CN109195975B - Diazaspiro [5.5] undecane derivative and application thereof - Google Patents

Diazaspiro [5.5] undecane derivative and application thereof Download PDF

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CN109195975B
CN109195975B CN201780033350.XA CN201780033350A CN109195975B CN 109195975 B CN109195975 B CN 109195975B CN 201780033350 A CN201780033350 A CN 201780033350A CN 109195975 B CN109195975 B CN 109195975B
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ethyl
ring
diazaspiro
oxa
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CN109195975A (en
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郑苏欣
张国彪
张晓波
李航
王文晶
杜勇
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Sichuan Haisco Pharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/08—Bronchodilators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

The invention relates to a diazaspiro [5.5]]Undecane derivatives and uses thereof, the derivatives are shown in the general formula (I) or stereoisomers, hydrates, metabolites, solvates, pharmaceutically acceptable salts, co-crystals or prodrugs thereof, the invention also relates to a preparation method of the derivatives and application in preparing drugs for treating airway obstruction diseases,

Description

Diazaspiro [5.5] undecane derivative and application thereof
Technical Field
The invention relates to a diazaspiro [5.5]]Undecane derivatives, their preparation method and their application in medicine, specifically a pharmaceutical composition having muscarinic receptor antagonism and beta-agonism2Novel diazaspiro [5.5] with dual activity for adrenergic receptor agonism]Undecane or a stereoisomer, hydrate, solvate, metabolite, pharmaceutically acceptable salt, co-crystal or prodrug thereof, a pharmaceutical composition thereof, and a use thereof in medicine.
Background
Bronchodilators play an important role in the treatment of respiratory diseases such as Chronic Obstructive Pulmonary Disease (COPD), asthma and the like. Bronchodilators widely used in the clinic include muscarinic receptor antagonists and beta 2-adrenergic agonists. Muscarinic receptor antagonists exert their effect on bronchodilation by lowering vagal cholinergic levels of airway smooth muscle. Currently used inhaled muscarinic receptor antagonists include ipratropium bromide, oxitropium bromide, glycopyrronium bromide, tiotropium bromide, aclidinium bromide and umeclidinium bromide. Beta is a2Adrenergic agonists reverse bronchoconstrictor responses to various mediators, such as acetylcholine, by stimulating adrenergic receptors on airway smooth muscle. Beta is currently used2Adrenergic agonists include salbutamol, salmeterol, arformoterol, formoterol, vilanterol and indacaterol. These drugs, in addition to improving lung function, may also improve the quality of life and reduce the exacerbation of the disease.
As more clinical findings were made, the combination was demonstratedMuscarinic receptor antagonists and beta2Adrenergic agonists are more effective than either therapeutic agent alone, and muscarinic receptor antagonists and β are now clinically used2Adrenergic agonists are prepared into compound preparations for treating asthma and severe COPD, and the compound preparations mainly comprise Anoro Ellipta (umeclidinium bromide/vilanterol), Ultibro Breezhaler (glycopyrronium bromide/indacaterol), ipratropium bromide/salbutamol and the like. Although the compound preparation has better treatment effect than the single preparation, the preparation of the preparation has higher requirements.
Therefore, it is desired to develop a compound having both muscarinic receptor antagonism and β2-an adrenergic agonist dual action drug having the pharmaceutical advantages of both component combinations while possessing a single molecular pharmacokinetics. These compounds are administered as a single therapeutic agent and provide bronchodilatory action from two distinct and possibly synergistic modes of action. In addition, have muscarinic receptor antagonism and beta2Adrenergic agonist dual action (MABA) compounds may also be combined with corticosteroid (ICS) anti-inflammatory agents drugs to form two therapeutic agents (MABA/ICS) to provide triple action therapeutic effects (Expert opin investig. drugs (2014)23 (4): 453-.
Therefore, there is a need to develop novel compounds having both muscarinic receptor antagonism and β2Dual active agents of adrenergic stimulation to provide more effective single therapeutic doses or compound formulations, providing more clinical medication options for the patient.
Disclosure of Invention
The invention provides a compound shown in a general formula (I) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof:
Figure GPA0000261095610000031
Wherein:
R1is selected from C6-12Carbocyclic ring or 5 toA 12 membered heterocyclic ring, said carbocyclic or heterocyclic ring optionally further substituted with 0 to 5R1aSubstituted and said heterocycle contains 1 to 4 heteroatoms selected from N, O or S;
R1aselected from the group consisting of F, Cl, Br, I, - (═ O), nitro, cyano, hydroxy, carboxy, -C (═ O) OC1-6Alkyl, amino, C1-6Alkyl radical, C2-6Alkynyl, C1-6Alkoxy radical, C3-12Carbocyclic, 5-to 12-membered heterocyclic, -O-C3-12Carbocyclic ring, -NH-C3-12Carbocyclic ring or-CH2-C3-6Cycloalkyl, said hydroxy, amino, alkyl, alkynyl, alkoxy, carbocycle, heterocycle or cycloalkyl optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, cyano, hydroxy, C1-6Alkyl radical, C2-6Alkynyl, C1-4Alkoxy radical, C3-6Cycloalkyl or C3-10Carbocyclic or 5 to 10 membered heterocyclic ring, said heterocyclic ring containing 1 to 4 heteroatoms selected from N, O or S;
w is selected from a bond or-C (═ O) -;
provided that when W is selected from-C (═ O) -, R1By 1 to 5R1aSubstituted and has at least 1R1aIs selected from C2-6Alkynyl or-CH2-C3-6A cycloalkyl group;
R2、R3、R4、R5each independently selected from C1-6Alkylene optionally further substituted by 0 to 4 groups selected from F, Cl, Br, I, cyano, hydroxy, C1-6Alkyl radical, C1-6Alkoxy, phenyl or phenyl-C1-4Substituted by a substituent of alkylene;
A is selected from C3-12A carbocyclic ring or a 5 to 12 membered heterocyclic ring, said carbocyclic or heterocyclic ring optionally further substituted by 0 to 5 of F, Cl, Br, I, cyano, hydroxy, amino, C1-6Alkyl radical, C2-6Alkynyl, C1-4Alkoxy or C3-6Cycloalkyl, and said heterocycle contains 1 to 3 heteroatoms selected from N, O or S;
X1and X2Each independently selected from the group consisting of a bond, -O-, -C (═ O) O-, -OC (═ O) -, -S-, -S(＝O)-、-S(＝O)2-、-C(＝O)NRx-、-NRxC(＝O)-、-OC(＝O)NRx-、-NRxC(＝O)O-、-NRxC(＝O)NRx、-NRxS(＝O)2-、-S(＝O)2NRx-、-NRxS(＝O)2NRxor-NRx-；
RxEach independently selected from H, C1-6Alkyl or C3-8Cycloalkyl, said alkyl or cycloalkyl being optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I, C1-4Alkyl radical, C3-6Cycloalkyl or C1-4Substituted by a substituent of alkoxy;
as alternative R3-X1-R4-X2-R5May be an ethylene group;
R6、R7each independently selected from H or C1-4An alkyl group;
R8selected from H or OH;
Figure GPA0000261095610000041
represents a beta-adrenoceptor binding group.
In a preferred embodiment of the present invention, a compound represented by the general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
Figure GPA0000261095610000042
represents a beta-adrenoceptor binding group;
b is preferably
Figure GPA0000261095610000043
R10、R11、R12、R13、R14、R15、R16、R17Or R18Each independently selected from H, F, Cl, Br, I, CF3、OH、-CH2OH, cyano, carboxyl, C1-4Alkyl radical, C1-4Alkoxy, -C (═ O) C1-4Alkyl, -C (═ O) OC 1-4Alkyl, -NHC (═ O) H, -NHS (═ O)2-C1-4Alkyl, -NHS (═ O)2-NH2or-NHS (═ O)2-NHC1-4Alkyl, Q is selected from-CRq1＝CRq2-、-CRq1Rq2CRq3Rq4-、-O-、-S-、-OCRq1Rq2-、-CRq1Rq2O-、-SCRq1Rq2-、-CRq1Rq2S-, said R-q1、Rq2、Rq3Or Rq4Each independently selected from H, F, Cl, Br, I or C1-4An alkyl group;
b is more preferably
Figure GPA0000261095610000051
Wherein Q is selected from-CH ═ CH-, -CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-；
B is further preferably
Figure GPA0000261095610000052
Figure GPA0000261095610000053
In a preferred embodiment of the present invention, a compound represented by the general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
R1is selected from C6-10A carbocyclic ring or a 5-to 10-membered heterocyclic ring, preferably a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring, a pyrazole ring, a pyridine ring, a quinoline ring, an isoquinoline ring, a benzisothiazole ring, an indazole ring, a 1H-pyrazole [4, 3-b ]]A pyridine ring or
Figure GPA0000261095610000054
The carbocycle, the heterocycle, the benzene ring, the thiophene ring,Furan ring, pyrrole ring, thiazole ring, oxazole ring, imidazole ring, isothiazole ring, isoxazole ring, pyrazole ring, pyridine ring, quinoline ring, isoquinoline ring, benzisothiazole ring, indazole ring, 1H-pyrazole [4, 3-b ]]A pyridine ring or
Figure GPA0000261095610000055
Optionally further substituted by 0 to 5R1aSubstituted and said heterocycle contains 1 to 4 heteroatoms selected from N, O or S;
R1aSelected from the group consisting of F, Cl, Br, - (═ O), nitro, cyano, hydroxy, carboxy, -C (═ O) OC1-4Alkyl, amino, C1-4Alkyl radical, C2-4Alkynyl, C1-4Alkoxy radical, C3-10Carbocyclic, 5-to 10-membered heterocyclic, -O-C3-10Carbocyclic ring, -NH-C3-10Carbocyclic ring or-CH2-C3-6Cycloalkyl, said hydroxy, amino, alkyl, alkynyl, alkoxy, carbocycle, heterocycle or cycloalkyl optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, cyano, hydroxy, C1-4Alkyl radical, C1-4Alkoxy radical, C3-6Cycloalkyl radical, C3-10Carbocyclic or 5 to 10 membered heterocyclic ring, said heterocyclic ring containing 1 to 4 heteroatoms selected from N, O or S;
w is selected from a bond or-C (═ O) -;
provided that when W is selected from-C (═ O) -, R1By 1 to 5R1aSubstituted and has at least 1R1aIs selected from C2-4Alkynyl or-CH2-C3-6A cycloalkyl group;
R2、R5each independently selected from C1-4Alkylene optionally further substituted with 0 to 4 substituents selected from F, methyl, ethyl, methoxy or ethoxy;
R3、R4each independently selected from C1-4Alkylene, preferably methylene, ethylene, propylene, -CH2CH(CH3)-、-CH(CH3)CH2-、-C(CH3)2CH2-、-CH2C(CH3)2-, butylene, -CH (CH)3)CH2CH2-、-CH2CH(CH3)CH2-or-CH2CH(CH3)CH2-said alkylene, methylene, ethylene, propylene or butylene is optionally further substituted with 0 to 4 substituents selected from F, methyl, ethyl, propyl, isopropyl, butyl, methoxy, ethoxy, phenyl-methylene, phenyl-ethylene, phenyl-propylene or phenyl-butylene;
A is selected from C6-10Carbocyclic or 5 to 10 membered heterocyclic ring, preferably
Figure GPA0000261095610000061
The carbocycle, the heterocycle,
Figure GPA0000261095610000062
Figure GPA0000261095610000063
Optionally further substituted by 0 to 5 of F, Cl, Br, I, cyano, hydroxy, amino, C1-6Alkyl radical, C2-6Alkynyl, C1-4Alkoxy or C3-6Cycloalkyl, and said heterocycle contains 1 to 3 heteroatoms selected from N, O or S;
X1and X2Each independently selected from the group consisting of a bond, -O-, -C (O) O-, -OC (O) -, -S (O)2-、-C(＝O)NRx-、-NRxC(＝O)-、-OC(＝O)NRx-、-NRxC(＝O)O-、-NRxC(＝O)NRx、-NRxS(＝O)2-、-S(＝O)2NRx-、-NRxS(＝O)2NRxor-NRx-, preferably a bond, -O-, -C (═ O) -, -C (═ O) O-, -OC (═ O) -, -C (═ O) NRx-、-NRxC(＝O)-、-OC(＝O)NRx-、-NRxC (═ O) O-or-NRx-；
RxEach independently selected from H, C1-6Alkyl or C3-8Cycloalkyl, said alkyl or cycloalkyl being optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I, methyl, ethyl,Cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy or ethoxy;
Rxpreferably H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
as alternative R3-X1-R4-X2-R5May be an ethylene group;
R6、R7each independently selected from H or C1-4Alkyl, preferably H, methyl or ethyl;
R8selected from H or OH;
b is selected from
Figure GPA0000261095610000064
Wherein Q is selected from-CH ═ CH-, -CH 2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-；
B is preferably
Figure GPA0000261095610000065
Figure GPA0000261095610000066
In a preferred embodiment of the present invention, a compound represented by the general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
R1selected from the group consisting of a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring, a pyrazole ring, a pyridine ring, a pyrimidine ring, a pyrazine ring, a quinoline ring, an isoquinoline ring, a benzisothiazole ring, an indazole ring, a 1H-pyrazolo [4, 3-b ] ring]A pyridine ring or
Figure GPA0000261095610000067
The benzene ring, the thiophene ring and the furan ringPyrrole ring, thiazole ring, oxazole ring, imidazole ring, isothiazole ring, isoxazole ring, pyrazole ring, pyridine ring, pyrimidine ring, pyrazine ring, quinoline ring, isoquinoline ring, benzisothiazole ring, indazole ring, 1H-pyrazole [4, 3-b ]]A pyridine ring or
Figure GPA0000261095610000071
Optionally further substituted by 0 to 5 substituents selected from R1aSubstitution;
R1aselected from F, Cl, Br, nitro, amino, cyano, carboxyl, -C (═ O) OBn, -CF3Hydroxy, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, ethynyl, propynyl, propargyl, methoxy, ethoxy, propoxy, isopropoxy, hydroxymethyl, hydroxyethyl, -C (CH) 3)2OH, phenoxy, anilino, phenyl, 2-thienyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or-CH2-a cyclopropyl group;
w is selected from a bond or-C (═ O) -;
provided that when W is selected from-C (═ O) -, R1By 1 to 5R1aSubstituted and has at least 1R1aSelected from ethynyl, propynyl, propargyl or-CH2-a cyclopropyl group;
R2、R5each independently selected from C1-4Alkylene optionally further substituted with 0 to 4 substituents selected from F, methyl, ethyl, methoxy or ethoxy;
R2、R5preferably methylene, ethylene, propylene, -CH2CH(CH3)-、-CH(CH3)CH2-、-C(CH3)2CH2-、-CH2C(CH3)2-, butylene, -CH (CH)3)CH2CH2-、-CH2CH(CH3)CH2-or-CH2CH(CH3)CH2-；
R3、R4Each independently selected from methylene, ethylene, propylene or butylene, said methylene, ethylene, propylene or butylene being optionally further substituted0 to 4 substituents selected from F, methyl, ethyl, propyl, isopropyl, butyl, methoxy, ethoxy, phenyl-methylene, phenyl-ethylene, phenyl-propylene or phenyl-butylene;
a is selected from
Figure GPA0000261095610000072
Said
Figure GPA0000261095610000073
Optionally further substituted with 0 to 5 substituents of F, Cl, Br, I, cyano, hydroxy, amino, methyl, ethyl, ethynyl, methoxy, ethoxy, cyclopropyl or cyclobutyl;
X1And X2Each independently selected from the group consisting of a bond, -O-, -C (═ O) O-, -OC (═ O) -, -C (═ O) NRx-、-NRxC(＝O)-、-OC(＝O)NRx-、-NRxC (═ O) O-or-NRx-；
RxEach independently selected from H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
as alternative R3-X1-R4-X2-R5May be an ethylene group;
R6、R7each independently selected from H, methyl or ethyl;
b is selected from
Figure GPA0000261095610000074
Figure GPA0000261095610000075
In a preferred embodiment of the present invention, the compound represented by the general formula (II) or (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
Figure GPA0000261095610000081
in formula (II) or formula (III):
p1 is selected from 0 or 1;
p2 is selected from 0, 1, 2, 3 or 4;
Raselected from H, F, Cl, Br, I, C1-4Alkyl radical, C1-4Alkoxy or C3-6A cycloalkyl group;
R1is selected from C6-10A carbocyclic ring or a 5 to 10 membered heterocyclic ring optionally further substituted with 0 to 5R1aSubstituted and said heterocycle contains 1 to 4 heteroatoms selected from N, O or S;
R1aselected from the group consisting of F, Cl, Br, - (═ O), nitro, cyano, hydroxy, amino, carboxy, -C (═ O) OC1-4Alkyl radical, C1-4Alkyl radical, C2-4Alkynyl, C1-4Alkoxy radical, C3-10Carbocyclic, 5-to 10-membered heterocyclic, -O-C 3-10Carbocyclic ring, -NH-C3-10Carbocyclic ring or-CH2-C3-6Cycloalkyl, said hydroxy, amino, alkyl, alkynyl, alkoxy, carbocycle, heterocycle or cycloalkyl optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, cyano, hydroxy, C1-4Alkyl radical, C1-4Alkoxy radical, C3-6Cycloalkyl radical, C3-10Carbocyclic or 5 to 10 membered heterocyclic ring, said heterocyclic ring containing 1 to 4 heteroatoms selected from N, O or S;
w is selected from a bond or-C (═ O) -;
provided that when W is selected from-C (═ O) -, R1By 1 to 5R1aSubstituted and has at least 1R1aIs selected from C2-4Alkynyl or-CH2-C3-6A cycloalkyl group;
Rxeach independently selected from H, C1-6Alkyl or C3-8Cycloalkyl, said alkyl or cycloalkyl optionally further substituted with 0 to 5 substituents selected from F, Cl, Br, I, methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy or ethoxy;
Rxpreferably H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
R8selected from H or hydroxy;
b is selected from
Figure GPA0000261095610000082
Q is selected from-CH ═ CH-, -CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-；
B is preferably
Figure GPA0000261095610000091
Figure GPA0000261095610000092
In a preferred embodiment of the present invention, the compound represented by the general formula (II) or (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
p1 is selected from 0 or 1;
p2 is selected from 0, 1, 2, 3 or 4;
R1selected from the group consisting of a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring, a pyrazole ring, a pyridine ring, a pyrazine ring, a pyrimidine ring, a quinoline ring, an isoquinoline ring, a benzisothiazole ring, an indazole ring, a 1H-pyrazolo [4, 3-b ] ring]A pyridine ring or
Figure GPA0000261095610000093
The benzene ring, the thiophene ring, the furan ring, the pyrrole ring, the thiazole ring, the oxazole ring, the imidazole ring, the isothiazole ring, the isoxazole ring, the pyrazole ring, the pyridine ring, the pyrimidine ring, the pyrazine ring, the quinoline ring, the isoquinoline ring, the benzisothiazole ring, the indazole ring, the 1H-pyrazole [4, 3-b ]]A pyridine ring or
Figure GPA0000261095610000094
Optionally further substituted by 0 to 5R1aSubstitution;
R1aselected from F, Cl, Br, nitro, amino, cyano, hydroxy, carboxy, -C (═ O) OBn, -CF3Methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, ethynyl, propynyl, propargyl, methoxy, ethoxy, propoxy, isopropoxy, hydroxymethyl, hydroxyethyl, -C (CH)3)2OH, phenoxy, anilino, phenyl, 2-thienyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or-CH2-a cyclopropyl group;
provided that when W is selected from-C (═ O) -, R 1By 1 to 5R1aSubstituted and has at least 1R1aSelected from ethynyl, propynyl, propargyl, or-CH2-a cyclopropyl group;
Raselected from F, Cl, Br, I, methyl, ethyl, methoxy, ethoxy or cyclopropyl;
Rxselected from H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
b is selected from
Figure GPA0000261095610000095
In a preferred embodiment of the present invention, the compound represented by the general formula (II) or (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:
p1 is selected from 0 or 1;
p2 is selected from 0, 1, 2, 3 or 4;
R1-w is selected from
Figure GPA0000261095610000101
Figure GPA0000261095610000102
RaSelected from F, Cl, Br, I, methyl, ethyl, methoxy, ethoxy or cyclopropyl;
Rxselected from H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
b is selected from
Figure GPA0000261095610000103
In a preferred embodiment of the present invention, the compound of the present invention includes, but is not limited to, one of the following compounds:
Figure GPA0000261095610000104
Figure GPA0000261095610000111
Figure GPA0000261095610000121
Figure GPA0000261095610000131
Figure GPA0000261095610000141
Figure GPA0000261095610000151
Figure GPA0000261095610000161
Figure GPA0000261095610000171
Figure GPA0000261095610000181
the invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of the general formulae (I), (II) or (III), or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, in combination with a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more other therapeutic agents; preferably wherein the other therapeutic agent is selected from one or more of a PDE4 inhibitor, an M receptor antagonist, a corticosteroid, and a β -adrenoceptor agonist.
The invention also relates to application of the compound shown in the general formula (I), (II) or (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic crystal or prodrug thereof, or the pharmaceutical composition in preparing medicaments for treating airway obstructive diseases, preferably asthma, chronic obstructive pulmonary disease or bronchitis.
The present invention also provides a method for treating an obstructive disease of the airways comprising administering a compound of formula (I), (II) or (III) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, or a pharmaceutical composition thereof, preferably the obstructive disease of the airways is selected from asthma, chronic obstructive pulmonary disease or bronchitis.
The invention also provides an intermediate for preparing the compound shown in the formula (I), wherein the intermediate is selected from the compounds shown in the formula (M-1), the formula (M-2) or the formula (M-3) or a stereoisomer and a pharmaceutically acceptable salt thereof:
Figure GPA0000261095610000182
Figure GPA0000261095610000191
wherein:
Rm1selected from amino protecting groups or H;
Rm2selected from-COOH, -COOC1-6Alkyl, -X2-R5-OH、-X2-R5-a leaving group, -X2-Rm3CHO、-X2-Rm3C(OC1-6Alkyl radical)2Or
Figure GPA0000261095610000192
Said alkyl group optionally being further substituted by 0 to 4 of F, Cl, Br or C1-4Alkyl substituted;
Rm3is selected from the group consisting of5Alkylene with one less carbon atom;
R1is selected from C6-12Carbocyclic or 5 to 12 membered heterocyclic ring, preferably C6-10A carbocyclic ring or a 5-to 10-membered heterocyclic ring, more preferably a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring, a pyrazole ring, a pyridine ring, a pyrimidine ring, a pyrazine ring, a quinoline ring, an isoquinoline ring, a benzisothiazole ring, an indazole ring, 1H-pyrazole [4, 3-b ] ring]A pyridine ring or
Figure GPA0000261095610000193
The carbocycle, heterocycle, benzene ring, thiophene ring, furan ring, pyrrole ring, thiazole ring, oxazole ring, imidazole ring, isothiazole ring, isoxazole ring, pyrazole ring, pyridine ring, pyrimidine ring, pyrazine ring, quinoline ring, isoquinoline ring, benzisothiazole ring, indazole ring, 1H-pyrazole [4, 3-b ]]A pyridine ring or
Figure GPA0000261095610000194
Optionally further substituted by 0 to 5R1aSubstituted and said heterocycle contains 1 to 4 heteroatoms selected from N, O or S;
R1aselected from the group consisting of F, Cl, Br, I, - (═ O), nitro, cyano, hydroxy, carboxy, -C (═ O) OC1-6Alkyl, amino, C1-6Alkyl radical, C2-6Alkynyl, C1-6Alkoxy radical, C3-12Carbocyclic, 5-to 12-membered heterocyclic, -O-C3-12Carbocyclic ring, -NH-C3-12Carbocyclic ring or-CH 2-C3-6Cycloalkyl, said hydroxy, amino, alkyl, alkynyl, alkoxy, carbocycle, heterocycle or cycloalkyl optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, cyano, hydroxy, C1-6Alkyl radical, C2-6Alkynyl, C1-4Alkoxy radical, C3-6Cycloalkyl or C3-10Carbocyclic or 5 to 10 membered heterocyclic ring, said heterocyclic ring containing 1 to 4 heteroatoms selected from N, O or S;
R1apreferred are F, Cl, Br, - (═ O), nitro, cyano, hydroxy, amino, carboxy, -C (═ O) OC1-4Alkyl radical, C1-4Alkyl radical, C2-4Alkynyl, C1-4Alkoxy radical, C3-10Carbocyclic, 5-to 10-membered heterocyclic, -O-C3-10Carbocyclic ring, -NH-C3-10Carbocyclic ring or-CH2-C3-6Cycloalkyl, said hydroxy, amino, alkyl, alkynyl, alkoxy, carbocycle, heterocycle or cycloalkyl optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, cyano, hydroxy, C1-4Alkyl radical, C1-4Alkoxy radical, C3-6Cycloalkyl radical, C3-10Carbocyclic or 5 to 10 membered heterocyclic ring, said heterocyclic ring containing 1 to 4 heteroatoms selected from N, O or S;
R2、R3、R4、R5each independently selected from C1-6Alkylene, preferably C1-4Alkylene optionally further substituted by 0 to 4 groups selected from F, Cl, Br, I, cyano, hydroxy, C1-6Alkyl radical, C1-6Alkoxy, phenyl or phenyl-C1-4Substitution of alkylene groups Substituted by a group;
R2、R5more preferably methylene, ethylene, propylene, -CH2CH(CH3)-、-CH(CH3)CH2-、-C(CH3)2CH2-、-CH2C(CH3)2-, butylene, -CH (CH)3)CH2CH2-、-CH2CH(CH3)CH2-or-CH2CH(CH3)CH2-；
R3、R4More preferably methylene, ethylene, propylene or butylene;
a is selected from C3-12Carbocyclic or 5 to 12 membered heterocyclic ring, preferably C6-10A carbocyclic ring or a 5 to 10 membered heterocyclic ring, said carbocyclic or heterocyclic ring optionally further substituted by 0 to 5 of F, Cl, Br, I, cyano, hydroxy, amino, C1-6Alkyl radical, C2-6Alkynyl, C1-4Alkoxy or C3-6Cycloalkyl, and said heterocycle contains 1 to 3 heteroatoms selected from N, O or S;
X1and X2Each independently selected from the group consisting of a bond, -O-, -C (O) O-, -OC (O) -, -S (O)2-、-C(＝O)NRx-、-NRxC(＝O)-、-OC(＝O)NRx-、-NRxC(＝O)O-、-NRxC(＝O)NRx、-NRxS(＝O)2-、-S(＝O)2NRx-、-NRxS(＝O)2NRxor-NRx-；
RxEach independently selected from H, C1-6Alkyl or C3-8Cycloalkyl, said alkyl or cycloalkyl being optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I, C1-4Alkyl radical, C3-6Cycloalkyl or C1-4Substituted by a substituent of alkoxy;
Rxpreferably H, C1-6Alkyl or C3-8Cycloalkyl, said alkyl or cycloalkyl being optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I, methyl, ethyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy or ethoxySubstitution;
Rxmore preferably H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH 2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
the leaving group is selected from chlorine, bromine, iodine, -OMs or OTs;
n1 is selected from 1, 2 or 3;
with the proviso that formula (M-1) is not
Figure GPA0000261095610000201
The invention relates to a preferable scheme for preparing an intermediate of a compound shown as a formula (I), wherein the intermediate is selected from a compound shown as a formula (M-1), a formula (M-2) or a formula (M-3) or a stereoisomer and a pharmaceutically acceptable salt thereof,
R1-w is selected from
Figure GPA0000261095610000202
Figure GPA0000261095610000203
Figure GPA0000261095610000211
A is selected from
Figure GPA0000261095610000212
Said
Figure GPA0000261095610000213
Optionally further substituted with 0 to 5 substituents of F, Cl, Br, I, cyano, hydroxy, amino, methyl, ethyl, ethynyl, methoxy, ethoxy, cyclopropyl or cyclobutyl;
Rm1selected from amino protecting groups or H, preferably tert-butyloxycarbonyl or H;
Rm2selected from-COOH, -COOC1-6Alkyl, aryl, heteroaryl, and heteroaryl,-X2-R5-OH、-X2-R5-a leaving group, -X2-Rm3CHO、-X2-Rm3C(OC1-6Alkyl radical)2Or
Figure GPA0000261095610000214
Said alkyl group optionally being further substituted by 0 to 4 of F, Cl, Br or C1-4Alkyl substituted, Rm2preferably-COOH, -COOt-Bu or-X2-Rm3C(OCH3)2；
Rm3Is selected from the group consisting of5Alkylene having one less carbon atom, Rm3Preferably methylene or ethylene;
X1is selected from-O-;
X2selected from a bond or-C (═ O) NRx-；
RxSelected from H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH 2-a cyclopentyl group;
R2、R3、R4、R5each independently selected from methylene, ethylene or propylene;
the leaving group is selected from chlorine, bromine, iodine, -OMs or OTs;
n1 is selected from 1, 2 or 3.
The present invention is a preferred embodiment of an intermediate for preparing a compound represented by formula (I) which is a compound represented by the following structure or a stereoisomer and a pharmaceutically acceptable salt thereof:
Figure GPA0000261095610000215
Figure GPA0000261095610000221
Figure GPA0000261095610000231
Figure GPA0000261095610000241
Figure GPA0000261095610000251
the process for the preparation of the preferred compounds of the invention is optionally prepared by one of the following routes (the following reaction steps are partly prepared by the "one-pot" method):
Figure GPA0000261095610000252
Figure GPA0000261095610000261
R1-w is selected from
Figure GPA0000261095610000262
Figure GPA0000261095610000263
RxSelected from H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
b is selected from
Figure GPA0000261095610000264
RaSelected from H, F, Cl, Br, methyl, ethyl, isopropyl or CF3；
The compound A (or a salt thereof, preferably trifluoroacetate) and the compound B are subjected to substitution reaction under alkaline conditions to obtain a compound C, preferably under the conditions that a reaction substrate is dissolved in dioxane, acetonitrile, tetrahydrofuran or 2-methyltetrahydrofuran, and a small amount of water (3-5% (v/v) of a reaction solvent) can be added in the presence of 1-6 equivalents of sodium tert-butoxide, potassium phosphate, cesium carbonate or potassium carbonate, so that the reaction is performed at room temperature to reflux.
Hydrolyzing the compound C to generate a compound D (or a salt thereof, preferably trifluoroacetate salt), preferably, dissolving a reaction substrate in dichloromethane, adding trifluoroacetic acid (1/5-1/1 of dichloromethane volume), and reacting at room temperature.
And (3) carrying out substitution reaction on the compound E and the compound D (or a salt thereof, preferably trifluoroacetic acid salt) to obtain a compound F, preferably, dissolving a reaction substrate in dichloromethane, adding 1.0-3 equivalents of HATU in the presence of 1-6 equivalents of triethylamine, 4-dimethylaminopyridine or N, N-diisopropylethylamine, and reacting at room temperature.
And reacting the compound F under the aldehyde group removal protection condition to obtain a compound G, preferably dissolving a reaction substrate in tetrahydrofuran, adding 1-10 equivalents of acetic acid or p-toluenesulfonic acid monohydrate, and reacting at room temperature-50 ℃.
And carrying out reductive amination reaction on the compound G and a compound H1 to obtain a compound T1 (the preferable compound of the invention), wherein the preferable conditions are that a reaction substrate is dissolved in N-methylpyrrolidone or N, N-dimethylformamide, 0.8-2 equivalents of acetic acid is added, stirring is carried out at room temperature for 0.5-1 hour, 2-3 equivalents of sodium triacetoxyborohydride is added, and reaction is carried out at room temperature.
And (3) carrying out reductive amination reaction on the compound G and a compound H2 to obtain a compound I, preferably dissolving a reaction substrate in a mixed solvent of dichloromethane and methanol (preferably, v/v is 10/3-10/1), stirring for 0.5-1 hour, adding 2-3 equivalents of sodium triacetoxyborohydride, and reacting at room temperature.
The compound I is subjected to TBS removal protection to obtain a compound T2 (the preferable compound of the invention), and the preferable conditions are that a reaction substrate is dissolved in tetrahydrofuran, 5-20 equivalents of triethylamine trihydrofluoride are added, and the reaction is carried out at room temperature.
Compound C can also be reacted by the following pathway:
Figure GPA0000261095610000271
the compound A (or a salt thereof, preferably trifluoroacetate) and the compound J are subjected to substitution reaction under alkaline conditions to obtain a compound K, preferably under the conditions that a reaction substrate is dissolved in dioxane, acetonitrile, tetrahydrofuran or 2-methyltetrahydrofuran, and a small amount of water (3-5% (v/v) of a reaction solvent) can be added in the presence of 1-6 equivalents of sodium tert-butoxide, potassium phosphate, cesium carbonate or potassium carbonate, so that the reaction is performed at room temperature to reflux.
Alternatively, the compound A and the compound J with protected hydroxyl are subjected to substitution reaction under alkaline conditions and then subjected to reaction under dehydroxylation protection conditions to obtain the compound K, wherein the preferable conditions of the substitution reaction are that a reaction substrate is dissolved in dioxane, acetonitrile, tetrahydrofuran or 2-methyltetrahydrofuran, and a small amount of water (3-5% (v/v) of a reaction solvent) can be added in the presence of 1-6 equivalents of sodium tert-butoxide, potassium phosphate, cesium carbonate or potassium carbonate, and the reaction is performed at room temperature to reflux. When the hydroxyl protecting group is TBS, the substrate is dissolved in tetrahydrofuran, and TBAF (tetrabutylammonium fluoride) is added and reacted at room temperature.
And (3) reacting the compound K with tert-butyl acrylate to generate a compound C, preferably, dissolving a reaction substrate in acetonitrile, adding benzyltrimethylammonium hydroxide, and reacting at room temperature.
Compound K can also be obtained by reacting:
Figure GPA0000261095610000281
and (2) carrying out reductive amination reaction on the compound A (or salt thereof, preferably trifluoroacetate) and the compound N to obtain a compound K, preferably dissolving a reaction substrate in tetrahydrofuran, stirring at room temperature for 0.5-1 hour, adding 2-3 equivalents of sodium triacetoxyborohydride, and reacting at room temperature. Alternatively, compound a is subjected to a reductive amination reaction with a hydroxy-protected compound N, followed by deprotection to form compound K. When the hydroxyl protecting group is TBS, the substrate is dissolved in tetrahydrofuran, and TBAF (tetrabutylammonium fluoride) is added and reacted at room temperature.
Compound C can also be obtained by reacting:
Figure GPA0000261095610000282
and (2) carrying out reductive amination reaction on the compound A (or salt thereof, preferably trifluoroacetate) and the compound M to obtain a compound C, preferably dissolving a reaction substrate in tetrahydrofuran, stirring at room temperature for 0.5-1 hour, adding 2-3 equivalents of sodium triacetoxyborohydride, and reacting at room temperature.
Compound a (or a salt thereof, preferably the trifluoroacetate salt) can be obtained by a pathway reaction:
Figure GPA0000261095610000283
LG is selected from Cl, Br or I;
R1in accordance with the synthetic route described above;
PG is selected from tert-butyloxycarbonyl;
the compound A2 and the compound A4 are subjected to coupling reaction under alkaline conditions to generate a compound A1(W ═ bond), preferably under the conditions that a reaction substrate is dissolved in dioxane, toluene, dimethyl sulfoxide or N, N-dimethylformamide, and 0-0.2 equivalent of palladium acetate or tris (dibenzylideneacetone) dipalladium catalyst and optionally 0-0.2 equivalent of 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene, triphenylphosphine or tri-tert-butylphosphine ligand are added in the presence of 1-3 equivalents of sodium tert-butoxide, potassium phosphate, cesium carbonate or potassium carbonate to react at the temperature of 60-150 ℃;
or the compound a2 and the compound a4 are subjected to substitution reaction under an alkaline condition to produce a compound a1(W ═ bond), preferably under a condition that a reaction substrate is dissolved in acetonitrile, dioxane, toluene, dimethyl sulfoxide or N, N-dimethylformamide and reacted at room temperature to 150 ℃ in the presence of 0.8 to 3 equivalents of 1, 5-diazabicyclo [5.4.0] undec-5-ene, potassium carbonate, sodium carbonate, triethylamine, cesium carbonate or potassium carbonate.
Compound A3 and compound a4 are subjected to condensation reaction to produce compound a1(W ═ C ═ O), preferably under conditions in which the reaction substrate is dissolved in dichloromethane, 2 to 5 equivalents of triethylamine and 1 to 2 equivalents of an activating agent (preferably 1-propylphosphoric anhydride (T3P, CAS: 68957-94-8) and HATU (CAS: 148893-10-1 or carbonyldiimidazole) are added, and the reaction is carried out at room temperature.
Unless stated to the contrary, the terms used in the specification and claims have the following meanings.
Carbon, hydrogen, oxygen, sulfur, nitrogen or halogen referred to in the groups and compounds of the invention all include isotopes thereof, and carbon, hydrogen, oxygen, sulfur, nitrogen or halogen referred to in the groups and compounds of the invention are optionally further replaced by one or more of their corresponding isotopes, wherein isotopes of carbon include isotopes of carbon12C、13C and14c, isotopes of hydrogen including protium (H), deuterium (D, also known as deuterium), tritium (T, also known as deuterium), and isotopes of oxygen including16O、17O and18isotopes of O, sulfur including32S、33S、34S and36isotopes of S, nitrogen include14N and15isotopes of N, F19Isotopes of F, chlorine including35Cl and37cl, isotopes of bromine including79Br and81Br。
"alkyl" means a straight and branched chain monovalent saturated hydrocarbon group, the backbone comprising 1 to 10 carbon atoms, preferably 1 to 8 carbon atoms, further preferably 1 to 6 carbon atoms, more preferably 1 to 4 carbon atoms, straight and branched chain groups, most preferably 1 to 2 carbon atoms, examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, and the like; said alkyl may optionally be further substituted by 0 to 5 substituents selected from F, Cl, Br, I, ═ O, -CH 2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3Hydroxy, -SR19Nitro, cyano, isocyano, alkyl, hydroxyalkyl, alkoxy, carbocyclyl, heterocyclyl, C2-8Alkenyl radical, C2-8Alkynyl, - (CH)2)k-C(＝O)-R19、-(CH2)k-C(＝O)-O-R19、-(CH2)k-C(＝O)-NR19R19a、-(CH2)k-S(＝O)j-R19、-O-C(＝O)-O-R19or-NR19R19aWherein R is substituted by a substituent of (1)19And R19aEach independently selected from H, hydroxy, amino, carboxyl and C1-8Alkyl radical, C1-8Alkoxy radical, C2-8Alkenyl radical, C2-8Alkynyl, 3-to 10-membered carbocyclyl, 4-to 10-membered heterocyclyl, 3-to 10-membered carbocyclyloxy or 4-to 10-membered heterocyclyloxy, k is selected from 0, 1, 2, 3, 4 or 5, and j is selected from 0, 1 or 2. Alkyl, k, j or R as appearing herein18And R18aAs defined above.
"alkylene" refers to a straight and branched chain divalent saturated hydrocarbon radical, including- (CH)2)v- (v is an integer of 1 to 10), examples of alkylene include, but are not limited to, methylene, ethylene, propylene, butylene, and the like; said alkylene group may optionally be further substituted by 0 to 5 groups selected from F, Cl, Br, I, ═ O, -CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3Hydroxy, -SR19Nitro, cyano, isocyano, alkyl, hydroxyalkyl, alkoxy, carbocyclyl, heterocyclyl, C2-8Alkenyl radical, C2-8Alkynyl, - (CH)2)k-C(＝O)-R19、-(CH2)k-C(＝O)-O-R19、-(CH2)k-C(＝O)-NR19R19a、-(CH2)k-S(＝O)j-R19、-O-C(＝O)-O-R19or-NR19R19aWhen the number of the substituents in the alkylene group is 2 or more,the substituents may be fused together to form a cyclic structure. Alkylene, as used herein, is defined as above.
"alkoxy" refers to a monovalent radical of an O-alkyl group, where alkyl is as defined herein, and examples of alkoxy include, but are not limited to, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, 2-methyl-2-propoxy, 1-pentyloxy, 2-pentyloxy, 3-pentyloxy, 2-methyl-2-butoxy, 3-methyl-1-butoxy, 2-methyl-1-butoxy, and the like.
"alkenyl" means a straight and branched chain monovalent unsaturated hydrocarbon group having at least 1, and usually 1, 2 or 3 carbon double bonds, and the main chain includes 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms, and even more preferably 2 to 4 carbon atoms in the main chain, examples of alkenyl include, but are not limited to, vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-3-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 2-hexenyl, and the like, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1-octenyl, 3-octenyl, 1-nonenyl, 3-nonenyl, 1-decenyl, 4-decenyl, 1, 3-butadiene, 1, 3-pentadiene, 1, 4-hexadiene, and the like; said alkenyl may optionally be further substituted by 0 to 5 substituents selected from F, Cl, Br, I, ═ O, -CH 2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3Hydroxy, -SR19Nitro, cyano, isocyano, alkyl, hydroxyalkyl, alkoxy, carbocyclyl, heterocyclyl, C2-8Alkenyl radical, C2-8Alkynyl, - (CH)2)k-C(＝O)-R19、-(CH2)k-C(＝O)-O-R19、-(CH2)k-C(＝O)-NR19R19a、-(CH2)k-S(＝O)j-R19、-O-C(＝O)-O-R19or-NR19R19aSubstituted with the substituent(s). Alkenyl as used herein, is defined as above.
"alkynyl" refers to straight and branched chain monovalent unsaturated hydrocarbon radicals having at least 1, and typically 1, 2 or 3 carbon-carbon triple bonds, and the backbone includes 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms, and even more preferably 2 to 4 carbon atoms in the backbone, with examples of alkynyl including, but not limited to, ethynyl, 1-propynyl, 2-propynyl, butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 4-pentynyl, 3-pentynyl, 1-methyl-2-butynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octynyl, 3-nonynyl, and 4-decynyl, and the like; said alkynyl may optionally be further substituted by 0 to 5 substituents selected from F, Cl, Br, I, ═ O, -CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3Hydroxy, -SR19Nitro, cyano, isocyano, alkyl, hydroxyalkyl, alkoxy, carbocyclyl, heterocyclyl, C2-8Alkenyl radical, C2-8Alkynyl, - (CH)2)k-C(＝O)-R19、-(CH2)k-C(＝O)-O-R19、-(CH2)k-C(＝O)-NR19R19a、-(CH2)k-S(＝O)j-R19、-O-C(＝O)-O-R19or-NR19R19aSubstituted with the substituent(s). Alkynyl, as found herein, is defined as above.
"cycloalkyl" refers to a monovalent saturated carbocyclic hydrocarbon group, typically of 3 to 10 carbon atoms, non-limiting examples including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl, and the like. Said cycloalkyl may optionally be further substituted by 0 to 5 substituents selected from F, Cl, Br, I, ═ O, -CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3Hydroxy, -SR19Nitro, cyano, isocyano, alkyl, hydroxyalkyl, alkoxy, carbocyclyl, heterocyclyl, C2-8Alkenyl radical, C2-8Alkynyl, - (CH)2)k-C(＝O)-R19、-(CH2)k-C(＝O)-O-R19、-(CH2)k-C(＝O)-NR19R19a、-(CH2)k-S(＝O)j-R19、-O-C(＝O)-O-R19or-NR19R19aSubstituted with the substituent(s). Cycloalkyl as found herein, is as defined above.
"carbocycle" or "carbocyclyl" refers to a saturated or unsaturated aromatic or non-aromatic ring which may be a 3 to 10 membered monocyclic, 4 to 12 membered bicyclic or 10 to 15 membered tricyclic ring system to which the carbocyclyl may be attached a bridged or spiro ring, non-limiting examples of which include cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-alkenyl, 1-cyclopentyl-2-alkenyl, 1-cyclopentyl-3-alkenyl, cyclohexyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, phenyl, naphthyl, benzocyclopropenyl, 2, 3-dihydrobenzocyclopropenyl, cyclodecyl, cyclopentadienyl, or the like,
Figure GPA0000261095610000301
The carbocyclyl may optionally be further substituted with 0 to 5 substituents selected from F, Cl, Br, I, ═ O, -CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3Hydroxy, -SR19Nitro, cyano, isocyano, alkyl, hydroxyalkyl, alkoxy, carbocyclyl, heterocyclyl, C2-8Alkenyl radical, C2-8Alkynyl, - (CH)2)k-C(＝O)-R19、-(CH2)k-C(＝O)-O-R19、-(CH2)k-C(＝O)-NR19R19a、-(CH2)k-S(＝O)j-R19、-O-C(＝O)-O-R19or-NR19R19aSubstituted with the substituent(s). Carbocyclyl as appearing herein, is defined as above.
"heterocycle" or "heterocyclyl" refers to a saturated or unsaturated aromatic or non-aromatic ring which may be a 3-to 10-membered monocyclic, 4-to 12-membered bicyclic, or 10-to 10-membered aromatic ringA 15 membered tricyclic ring system and containing 1 to 4 heteroatoms selected from N, O or S, preferably a 3 to 10 membered heterocyclyl group, the optionally substituted N, S of which in the ring may be oxidized to various oxidation states. The heterocyclic group may be attached at a heteroatom or carbon atom to which the heterocyclic group may be attached a bridged ring or a spiro ring, non-limiting examples of which include epoxyethyl, epoxypropyl, aziridinyl, oxetanyl, azetidinyl, thietanyl, 1, 3-dioxolanyl, 1, 4-dioxolanyl, 1, 3-dioxanyl, azepinyl, oxepinyl, thiepinyl, oxazepinyl, diazepinyl, thiazepinyl, pyridyl, piperidyl, homopiperidinyl, furyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, piperidyl, pering, morpholinyl, thiomorpholinyl, thiaoxazolidyl, 1, 3-dithianyl, dihydrofuranyl, dihydropyranyl, dithiazolyl, and the like, Tetrahydrofuryl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, chromanyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1, 3-dioxolanyl, pyrazolinyl, dithianyl, dithienylalkyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 1, 2, 3, 4-tetrahydroisoquinolinyl, 1, 2, 3, 4-tetrahydroquinolinyl, benzotriazolyl, 3-azabicyclo [3.1.0 ]Hexyl, 3-azabicyclo [4.1.0]Heptyl, azabicyclo [2.2.2]Hexyl, 3H-indolylquinozinyl, N-pyridylurea, 1-dioxothiomorpholinyl, azabicyclo [3.2.1]Octyl, azabicyclo [5.2.0 ] groups]Nonoalkyl oxatricyclo [5.3.1.1 ]]Dodecyl, azaadamantyl and oxaspiro [3.3 ]]A heptalkyl group. Said heterocyclyl may optionally be further substituted by 0 to 5 substituents selected from F, Cl, Br, I, ═ O, -CH2F、-CHF2、-CF3、-OCH2F、-OCHF2、-OCF3Hydroxy, -SR19Nitro, cyano, isocyano, alkyl, hydroxyalkyl, alkoxy, carbocyclyl, heterocycleBase, C2-8Alkenyl radical, C2-8Alkynyl, - (CH)2)k-C(＝O)-R19、-(CH2)k-C(＝O)-O-R19、-(CH2)k-C(＝O)-NR19R19a、-(CH2)k-S(＝O)j-R19、-O-C(＝O)-O-R19or-NR19R19aSubstituted with the substituent(s). Heterocyclyl, as used herein, is defined as above.
-OTs is
Figure GPA0000261095610000311
-OMs is
Figure GPA0000261095610000312
Bn being benzyl, i.e. -CH2A phenyl group.
"beta-adrenergic receptor binding group" refers to a group capable of binding to a beta-adrenergic receptor; see, for example, the review article "beta-acquired receivers in Comprehensive medical Chemistry, 1990, B.E. Main, p187(Pergamon Press)". See also, for example, WO/2005092841, US/20050215542, WO/2005070872, WO/2006023460, WO/2006051373, WO/2006087315 and WO/2006032627. Non-limiting examples include
Figure GPA0000261095610000313
R6、R7Each independently selected from H or C1-4Alkyl radical, R8Selected from H or OH, B is selected from
Figure GPA0000261095610000314
Figure GPA0000261095610000315
Wherein Q is selected from-CH ═ CH-, -CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-。
"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. Such as: "alkyl optionally substituted with F" means that the alkyl group may, but need not, be substituted with F, and the description includes the case where the alkyl group is substituted with F and the case where the alkyl group is not substituted with F.
"pharmaceutical composition" means a mixture of one or more compounds described herein or a physiologically/pharmaceutically acceptable salt thereof with other ingredients, wherein the other ingredients comprise physiologically/pharmaceutically acceptable carriers and excipients.
"carrier" refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.
"excipient" refers to an inert substance added to a pharmaceutical composition to further depend on the administration of the compound. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and different types of starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like.
"prodrug" refers to a compound that can be converted under physiological conditions or by solvolysis to a compound of the invention that is biologically active. Prodrugs of the invention are prepared by modifying functional groups in compounds of the invention, which modifications may be removed by routine manipulation or in vivo, to yield the parent compound.
"stereoisomers" refers to isomers resulting from the different arrangement of atoms in a molecule, including cis, trans isomers, enantiomers and conformational isomers.
An "effective dose" refers to an amount of a compound that causes physiological or medical translation in a tissue, system, or subject that is sought, including an amount of the compound that is sufficient to prevent, or alleviate to some extent, one or more symptoms of the condition or disorder being treated when administered to a subject.
"solvates" refers to compounds of the invention or salts thereof, which also include stoichiometric or non-stoichiometric amounts of solvents bound by intermolecular non-covalent forces. When the solvent is water, it is a hydrate.
“IC50"half inhibitory concentration" means the concentration at which half of the maximum inhibitory effect is achieved.
Detailed Description
The following detailed description is provided for the purpose of illustrating the embodiments and the advantageous effects thereof, and is not intended to limit the scope of the present disclosure.
The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or (and) Mass Spectrometry (MS). NMR shift (. delta.) of 10-6The units in (ppm) are given. NMR was measured using (Bruker Avance III 400 and Bruker Avance 300) nuclear magnetic spectrometers in deuterated dimethyl sulfoxide (DMSO-d)6) Deuterated chloroform (CDCl)3) Deuterated methanol (CD)3OD), internal standard Tetramethylsilane (TMS). MS was measured by Agilent 6120B (ESI) and Agilent 6120B (APCI). HPLC was carried out using an Agilent 1260DAD high pressure liquid chromatograph (Zorbax SB-C18100X 4.6 mm). The thin layer chromatography silica gel plate adopts HSGF254 of tobacco yellow sea or GF254 of Qingdao, the specification of the silica gel plate used by Thin Layer Chromatography (TLC) is 0.15 mm-0.20 mm, and the specification of the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm. The column chromatography generally uses 200-300 mesh silica gel of the Tibet Huanghai silica gel as a carrier. Known starting materials for the present invention can be synthesized by or according to methods known in the art, or can be purchased from companies such as Tatan technology, Annaiji chemistry, Shanghai Demer, Chengdong chemical, Shaoshou chemical technology, and Bailingwei technology. The nitrogen atmosphere means that the reaction flask is connected with a nitrogen balloon with a volume of about 1L. The hydrogen atmosphere refers to a reaction flask connected with a hydrogen balloon with a volume of about 1L. The hydrogenation reaction was usually evacuated and charged with hydrogen and repeated 3 times. In the examples, the reaction was carried out under a nitrogen atmosphere without specific mention. In the examples, the solution means an aqueous solution unless otherwise specified. In the examples, the reaction temperature is room temperature, unless otherwise specified. The room temperature is the most suitable reaction temperature and is 20-30 ℃. In the examples, M is mol/L, unless otherwise specified. TBS is tert-butyldimethyl And (3) silicon base. Boc is tert-butyloxycarbonyl. Bn is benzyl. TsOH. H2O is p-toluenesulfonic acid monohydrate. DMF is N, N-dimethylformamide. THF is tetrahydrofuran. DCM is dichloromethane.
Figure GPA0000261095610000331
HATU: (2- (7-Benzotolyltriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate CAS: 148893-10-1).
Intermediate 1: 3- [2- [3- (2-bromoethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester
tert-butyl 3-[2-[3-(2-bromoethyl)phenyl]ethoxy]propanoate
Figure GPA0000261095610000332
The first step is as follows: 2- (3- (2- ((tert-butyldimethylsilyl) oxy) ethyl) phenyl) ethanol (1b)
2-(3-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)ethan-1-ol
Sodium hydride (4.3g, 107.0mmol, 60% w/w) was added to THF (100mL), a THF solution (150mL) of 2, 2' - (1, 3-phenylene) diethanol (1a) (18.0g, 108.0mmol) was added dropwise at 0 deg.C, and after completion of the addition, stirring was carried out at 0 deg.C for 30 minutes, tert-butyldimethylsilyl chloride (14.5g, 108.0mmol) was added dropwise, and the mixture was stirred at room temperature for 10 hours. The reaction was quenched with water (150mL), concentrated under reduced pressure to remove most of THF, extracted with ethyl acetate (200mL × 2), the combined organic phases were washed with saturated brine (100mL × 1), the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 4: 1) to give 1b (10.2g, 34% yield) as a yellow liquid.
1H NMR(400MHz，CDCl3)δ7.27-7.22(m，1H)，7.10-7.07(m，3H)，3.87-3.80(m，4H)，2.87-2.80(m，4H)，0.89(s，9H)，0.00(s，6H).
The second step is that: 3- [2- [3- [2- [ tert-butyl (dimethyl) silyl ] oxyethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (1c)
tert-butyl 3-[2-[3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]phenyl]ethoxy]propanoate
Dissolve 1b (10.2g, 36.4mmol) in acetonitrile (30mL), add tert-butyl acrylate (14.0g, 109.0mmol) and benzyltrimethylammonium hydroxide (4.6g, 10.9mmol, 40% solution in methanol) and stir at room temperature for 3 hours. The reaction mixture was directly concentrated under reduced pressure to remove most of the reaction solvent, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give 1c (12.0g, yield 81%) as a yellow liquid.
1H NMR(400MHz，CDCl3)δ7.20-7.16(m，1H)，7.06-7.02(m，3H)，3.81-3.77(m，2H)，3.70-3.60(m，4H)，2.84-2.77(m，4H)，2.50-2.46(m，2H)，1.44(s，9H)，0.87(s，9H)，-0.02(s，6H).
The third step: 3- [2- [3- [ 2-hydroxyethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (1d)
tert-butyl 3-[2-[3-(2-hydroxyethyl)phenyl]ethoxy]propanoate
1c (12.0g, 29.4mmol) was dissolved in THF (100mL), tetrabutylammonium fluoride (15.4g, 58.7mmol) was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was directly concentrated under reduced pressure to remove most of the reaction solvent, water (100mL) was added and extracted with ethyl acetate (200mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 3: 1) to give 1d (7.2g, yield 83%) as a yellow solid.
1H NMR(400MHz，CDCl3)δ7.24-7.20(m，1H)，7.09-7.05(m，3H)，3.86-3.83(m，2H)，3.70-3.63(m，4H)，2.87-2.82(m，4H)，2.49-2.46(m，2H)，1.43(s，9H).
The fourth step: 3- [2- [3- (2-bromoethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (intermediate 1)
tert-butyl 3-[2-[3-(2-bromoethyl)phenyl]ethoxy]propanoate
1d (7.2g, 24.5mmol) was dissolved in methylene chloride (100mL), and imidazole (3.3g, 49.0mmol), triphenylphosphine (9.6g, 37.0mmol) and carbon tetrabromide (12.0g, 37.0mmol) were added and stirred for 2 hours. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate (100mL), extracted, the aqueous phase extracted with dichloromethane (150mL × 1), the combined organic phases dried over anhydrous sodium sulfate, filtered, the filtrate concentrated under reduced pressure and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to afford intermediate 1 as a yellow solid (7.5g, 86% yield).
1H NMR(400MHz，CDCl3)δ7.23-7.21(m，1H)，7.12-7.10(m，1H)，7.06-7.04(m，2H)，3.70-3.63(m，4H)，3.58-3.54(m，2H)，3.13-3.12(m，2H)，2.87-2.82(m，2H)，2.50-2.46(m，2H)，1.44(s，9H。
Intermediate 2: 3- [2- [3- (bromomethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (intermediate 2)
tert-butyl 3-[2-[3-(bromomethyl)phenyl]ethoxy]propanoate
Figure GPA0000261095610000341
The first step is as follows: 3- [2- (3-bromophenyl) ethoxy ] propionic acid tert-butyl ester (2b)
tert-butyl 3-[2-(3-bromophenyl)ethoxy]propanoate
M-bromophenylethyl alcohol (2a) (80.4g, 0.4mol) was placed in acetonitrile (200mL), tert-butyl acrylate (76.9g, 0.6mol) was added, benzyltrimethylammonium hydroxide (50.2g, 0.12mol, 40% solution in methanol) was added dropwise, and the mixture was stirred at 40 ℃ for 5 hours. Most of the reaction solvent was removed by concentration under reduced pressure, and water (200mL) and diethylamine (50mL) were added and stirred at room temperature for 30 minutes. Ethyl acetate (500mL) was added to the reaction mixture for extraction, the aqueous phase was extracted with ethyl acetate (500mL × 1), the combined organic phases were washed with saturated brine (200mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 12: 1) to give 2b (94.0g, yield 71.4%) as a yellow liquid.
1H NMR(400MHz，CDCl3)δ7.36-7.29(m，2H)，7.15-7.10(m，2H)，3.68-3.61(m，4H)，2.84-2.81(m，2H)，2.48-2.45(t，3H)，1.43(s，9H).
The second step is that: 3- [2- (3-cyanophenyl) ethoxy ] propionic acid tert-butyl ester (2c)
tert-butyl 3-[2-(3-cyanophenyl)ethoxy]propanoate
Dissolve 2b (9.9g, 30.0mmol) in DMF (60mL), add cuprous cyanide (5.4g, 60.0mmol) and stir at 150 ℃ for 16 h. The reaction was cooled to room temperature, water (100mL) and ethyl acetate (100mL) were added, celite was filtered, the filter cake was washed with water (100mL × 1) and ethyl acetate (100mL × 1) in this order, the filtrate was extracted, the aqueous phase was extracted with ethyl acetate (200mL × 1), the combined organic phases were washed with saturated brine (100mL × 1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give 2c as a yellow liquid (7.2g, yield 87%).
1H NMR(400MHz，CDCl3)δ7.50-7.44(m，3H)，7.37-7.33(m，1H)，3.67-3.63(m，4H)，2.89-2.85(t，3H)，2.46-2.42(m，2H)，1.41(s，9H).
LCMS m/z＝298.1[M+23]。
The third step: 3- [2- (3-formylphenyl) ethoxy ] propionic acid tert-butyl ester (2d)
tert-butyl 3-[2-(3-formylphenyl)ethoxy]propanoate
Pyridine (120mL), acetic acid (80mL) and water (80mL) were added to 2c (35.2g, 128mmol), raney nickel (1.8g, 30mmol) and sodium hypophosphite (27.1g, 256mmol) were added, and the mixture was stirred at 45 ℃ for 3 hours. Celite was filtered, the cake was washed with water (200mL) and ethyl acetate (300mL) in this order, the reaction solvent was removed by concentration under reduced pressure, water (100mL) was added and extracted with ethyl acetate (200mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtration was performed, the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 3: 1) to give 2d (22.0g, yield 61.8%) as a yellow liquid.
LCMS m/z＝301.2[M+23]。
The fourth step: 3- [2- [ (3-hydroxymethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (2e)
tert-butyl 3-[2-[3-(hydroxymethyl)phenyl]ethoxy]propanoate
Dissolve 2d (22.0g, 79.1mmol) in methanol (200mL), add sodium borohydride (6.0g, 158.2mmol), and stir at room temperature for 2 hours. Water (100mL) was added, and the mixture was concentrated under reduced pressure to remove most of the methanol, extracted with ethyl acetate (300mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2: 1) to give 2e (18.0g, yield 81.2%) as a yellow liquid.
1H NMR(400MHz，CDCl3)δ7.28-7.23(m，2H)，7.19-7.17(m，1H)，7.14-7.12(m，1H)，4.65(s，2H)，3.69-3.64(m，4H)，2.89-2.85(t.2H)，2.48-2.45(t，2H)，1.43(s，9H).
The fifth step: 3- [2- [3- (bromomethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (intermediate 2)
tert-butyl 3-[2-[3-(bromomethyl)phenyl]ethoxy]propanoate
Dissolve 2e (12.0g, 42.8mmol) in DCM (200mL), add triphenylphosphine (13.5g, 51.4mmol), carbon tetrabromide (17.0g, 51.4mmol), and imidazole (5.8g, 85.6mmol) at 0 deg.C, and stir at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (150mL) was added, extraction was performed, the aqueous phase was extracted with DCM (200mL × 1), the combined organic phases were dried over anhydrous sodium sulfate, filtration was performed, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give intermediate 2(4.0g, yield 27.0%) as a yellow liquid.
Intermediate 3: 3- [2- [4- (2-bromoethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (intermediate 3)
tert-butyl 3-[2-[4-(2-bromoethyl)phenyl]ethoxy]propanoate
Figure GPA0000261095610000351
The first step is as follows: 2- (4- (2- ((tert-butyldimethylsilyl) oxy) ethyl) phenyl) ethanol (3b)
2-(4-(2-((tert-butyldimethylsilyl)oxy)ethyl)phenyl)ethan-1-ol
Sodium hydride (4.0g, 100.0mmol, 60% w/w) was placed in THF (100mL), a THF solution (150mL) of 2, 2' - (1, 3-phenylene) diethanol (3a) (16.6g, 100.0mmol) was added dropwise at 0 deg.C, stirred for 30 minutes at 0 deg.C, then tert-butyldimethylchlorosilane (15.0g, 100.0mmol) was added dropwise and stirred at room temperature for 10 hours. The reaction was quenched by dropwise addition of water (150mL), concentrated under reduced pressure to remove most of THF, extracted with ethyl acetate (200mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was chromatographed on a silica gel column (petroleum ether/ethyl acetate (v/v) ═ 4: 1) to give 3b as a yellow liquid (11.2g, 40% yield).
LCMS m/z＝303.3[M+23]。
The second step is that: 3- [2- [4- [2- [ tert-butyl (dimethyl) silyl ] oxyethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (3c)
tert-butyl 3-[2-[4-[2-[tert-butyl(dimethyl)silyl]oxyethyl]phenyl]ethoxy]propanoate
3b (11.2g, 40.0mmol) was dissolved in acetonitrile (30mL), tert-butyl acrylate (10.3g, 80.0mmol) and benzyltrimethylammonium hydroxide (5.0g, 12.0mmol, 40% solution in methanol) were added and stirred at room temperature for 3 hours. Most of the reaction solvent was removed by concentration under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give 3c as a yellow liquid (13.9g, yield 85%).
The third step: 3- [2- [4- [ 2-hydroxyethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (3d)
tert-butyl 3-[2-[4-(2-hydroxyethyl)phenyl]ethoxy]propanoate
3c (13.9g, 34.0mmol) was dissolved in THF (100mL), tetrabutylammonium fluoride (17.8g, 68.0mmol) was added, and the mixture was stirred at room temperature for 3 hours. Most of the reaction solvent was removed by concentration under reduced pressure, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtration was performed, the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 3: 1) to give 3d as a yellow solid (8.0g, yield 80%).
LCMS m/z＝317.1[M+23]。
The fourth step: 3- [2- [4- (2-bromoethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (intermediate 3)
tert-butyl 3-[2-[4-(2-bromoethyl)phenyl]ethoxy]propanoate
3d (8.0g, 27.2mmol) was dissolved in DCM (100mL), imidazole (3.7g, 54.4mmol), triphenylphosphine (8.6g, 32.6mmol), carbon tetrabromide (10.8g, 32.6mmol) were added, a saturated aqueous sodium bicarbonate solution (100mL) was added with stirring at room temperature for 2 hours, extraction was performed, the aqueous phase was extracted with DCM (150 mL. times.1), the combined organic phases were dried over anhydrous sodium sulfate, filtration was performed, the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give intermediate 3(7.8g, yield 80%) as a yellow liquid.
LCMS m/z＝379.1[M+23]。
Intermediate 4: 3- [2- [4- (chloromethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (intermediate 4)
tert-butyl 3-[2-[3-(chloromethyl)phenyl]ethoxy]propanoate
Figure GPA0000261095610000361
The first step is as follows: 3- [2- (4-bromophenyl) ethoxy ] propionic acid tert-butyl ester (4b)
tert-butyl 3-[2-(4-bromophenyl)ethoxy]propanoate
4a (50.3g, 0.25mol) was taken up in acetonitrile (100mL), tert-butyl acrylate (48.1g, 0.375mol) was added and after that benzyltrimethylammonium hydroxide (31.4g, 0.075mol, 40% solution in methanol) was added dropwise and stirred for 5 h at 40 ℃. Most of the reaction solvent was removed by concentration under reduced pressure, and water (200mL) and diethylamine (50mL) were added and stirred at room temperature for 30 minutes. The reaction mixture was extracted with ethyl acetate (500mL × 2), the combined organic phases were washed with saturated brine (200mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (petroleum ether/ethyl acetate (v/v) ═ 12: 1) to give 4b (61.7g, yield 75%) as a yellow liquid.
The second step is that: 3- [2- (4-cyanophenyl) ethoxy ] propionic acid tert-butyl ester (4c)
tert-butyl 3-[2-(4-cyanophenyl)ethoxy]propanoate
4b (9.9g, 30.0mmol) was dissolved in N, N-dimethylformamide (60mL), cuprous cyanide (5.4g, 60.0mmol) was added, and the mixture was stirred at 150 ℃ for 16 hours. The reaction was cooled to room temperature, water (100mL) and ethyl acetate (100mL) were added, the mixture was filtered through celite, the cake was washed with water (100mL) and ethyl acetate (100mL) in this order, the filtrate was extracted, the aqueous phase was extracted with ethyl acetate (200mL × 1), the combined organic phases were washed with saturated brine (100mL × 1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed through a silica gel column (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give 4c as a yellow liquid (7.2g, yield 87%).
LCMS m/z＝298.1[M+23]。
The third step: 3- [2- (4-formylphenyl) ethoxy ] propionic acid tert-butyl ester (4d)
tert-butyl 3-[2-(4-formylphenyl)ethoxy]propanoate
4c (35.2g, 128mmol) was added to a mixture of pyridine (120mL), acetic acid (80mL) and water (80mL), Raney nickel (1.8g, 30mmol) and sodium hypophosphite (27.1g, 256mmol) were added, and the mixture was stirred at 45 ℃ for 3 hours. Celite was filtered, the cake was washed with water (200mL) and ethyl acetate (300mL) in this order, the filtrate was concentrated under reduced pressure to remove most of the reaction solvent, water (100mL) and ethyl acetate (200mL) were added to the residue, extraction was performed, the aqueous phase was extracted with ethyl acetate (200mL × 1), the combined organic phases were dried over anhydrous sodium sulfate, filtration was performed, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (petroleum ether/ethyl acetate (v/v) ═ 3: 1) to give 4d (22.0g, yield 61.8%) as a yellow liquid.
The fourth step: 3- [2- [ (4-hydroxymethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (4e)
tert-butyl 3-[2-[4-(hydroxymethyl)phenyl]ethoxy]propanoate
4d (22.0g, 79.1mmol) was dissolved in methanol (200mL), and sodium borohydride (6.0g, 158.2mmol) was added and stirred for 2 hours. Water (100mL) was added, and the mixture was concentrated under reduced pressure to remove most of the methanol, extracted with ethyl acetate (300mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2: 1) to give 4e (18.0g, yield 81.2%) as a yellow liquid.
1H NMR(400MHz，CDCl3)δ7.33-7.28(m，2H)，7.21-7.19(m，2H)，4.64(s，2H)，3.67-3.64(m，4H)，2.89-2.85(t，2H)，2.48-2.45(t，2H)，1.43(s，9H).
The fifth step: 3- [2- [4- (chloromethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester (intermediate 4)
tert-butyl 3-[2-[4-(chloromethyl)phenyl]ethoxy]propanoate
4e (11.2g, 40.0mmol) was dissolved in DCM (150mL), triethylamine (10.1g, 100.0mmol) and 4-dimethylaminopyridine (0.24g, 2.0mmol) were added at 0 deg.C, methanesulfonyl chloride (6.9g, 60.0mmol) was added dropwise, and the mixture was stirred at room temperature for 48 hours. Saturated aqueous sodium bicarbonate (150mL) was added, extraction was performed, the aqueous phase was extracted with DCM (200mL × 1), the combined organic phases were dried over anhydrous sodium sulfate, filtration was performed, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give intermediate 4(7.2g, yield 60.0%) as a yellow liquid.
LCMS m/z＝321.0[M+23]。
Intermediate 5: n- (2, 2-Dimethoxyethyl) butyl-1-amine (intermediate 5)
N-(2，2-dimethoxyethyl)butan-1-amine
Figure GPA0000261095610000381
After n-butylamine (5a) (14.6g, 0.2mol) was added to methanol (150mL) and aqueous glyoxal-1, 1-dimethylacetal solution (5b) (43.2g, 0.22mol, 60% aqueous solution) was added thereto, the mixture was stirred at room temperature for 6 hours, and then sodium borohydride (11.3g, 0.3mol) was added in portions and reacted at room temperature for 2 hours. After concentration under reduced pressure, most of the reaction solvent was removed, water (200mL) was added, and extraction was performed with DCM (300 mL. times.2), and after the organic phases were combined, they were dried over anhydrous sodium sulfate, and after concentration under reduced pressure, intermediate 5 was obtained as a yellow liquid (30.0g, yield 93%).
1H NMR(400MHz，CDCl3)δ4.45-4.42(m，1H)，3.35(s，6H)，2.70-2.69(m，2H)，2.60-2.56(m，2H)，1.44-1.40(m，2H)，1.33-1.26(m，2H)，0.90-0.86(m，3H)。
LCMS m/z＝162.2[M+1]。
Intermediate 6: 3- [2- [3- (chloromethyl) phenyl ] ethoxy ] propionic acid tert-butyl ester
tert-butyl 3-[2-[3-(chloromethyl)phenyl]ethoxy]propanoate
Figure GPA0000261095610000382
2e (91.0g, 324.5mmol) was added to DMF (450mL), N-dimethylpyridin-4-amine (3.97g, 32.5mmol) and triethylamine (110mL, 811.5mmol) were added, and after stirring at 0 ℃ for 5 minutes, methanesulfonyl chloride (37.7mL, 486.8mmol) was slowly added dropwise, and the reaction was carried out at room temperature for 8 hours. Saturated aqueous sodium bicarbonate (100mL) was added, extraction was performed with DCM (200mL × 3), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate (v/v) ═ 1: 0 to 8: 1) to give intermediate 6(54.5g, 56% yield) as a yellow liquid.
LCMS m/z＝321.1[M+23].
Intermediate 7: 3- [2- [3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] -2-fluorophenyl ] ethoxy ] acrylic acid tert-butyl ester
tert-butyl 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]ethoxy]propanoate
Figure GPA0000261095610000383
Figure GPA0000261095610000391
The first step is as follows: tert-butyl [2- (2-fluorophenyl) ethoxy ] dimethylsilane (7b)
tert-butyl-[2-(2-fluorophenyl)ethoxy]-dimethyl-silane
7a (21.03g, 150.0mmol) was added to DMF (150mL), imidazole (30.64g, 450.1mmol) and t-butyldimethylsilyl chloride (24.88g, 165.0mmol) were added at 0 deg.C, and after 10min, the reaction was allowed to warm to room temperature for 3.5 h. Water (200mL) was added, and extraction was performed with ethyl acetate (200 mL. times.2), and the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 7b as a colorless liquid (35.2g, yield: 92%).
The second step is that: 3- [2- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -2-fluorobenzaldehyde (7c)
3-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-2-fluoro-benzaldehyde
2, 2, 6, 6-tetramethylpyridine (39.1, 277mmol) was added to THF (200mL), n-butyllithium (0.133L, 160g/L) was slowly added dropwise at-78 deg.C, and after stirring for 15min, a solution of freshly 7b (35.2, 138mmol) in THF (100mL) was added dropwise and stirred at-78 deg.C for 2 h. A mixed solution of DMF (31mL) and THF (50mL) was added, and the mixture was stirred at-78 ℃ for 1 hour and then reacted at room temperature for 18 hours. The reaction solution was poured into 0.5M aqueous hydrochloric acid (1000mL), and 3.0M aqueous hydrochloric acid was added dropwise to a pH of about 7, followed by extraction with ethyl acetate (300 mL. times.3), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 7c (38.6g, yield: 99%) as a deep red liquid.
1H NMR(400MHz，CDCl3)δ10.36(s，1H)，7.74-7.69(m，1H)，7.53-7.48(m，1H)，7.23-7.13(m，1H)，7.06-6.95(m，1H)，3.84(t，2H)，2.94-2.89(m，2H)，0.84(d，9H)，-0.04(d，6H).
The third step: tert-butyl- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undec-9-yl ] methyl ] -2-fluorophenyl ] ethoxy ] -dimethylsilane (7d)
tert-butyl-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]ethoxy]-dimethyl-silane
Tert-butyl 4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (16C) (1.80g, 4.98mmol) was added to DCM (20mL), trifluoroacetic acid (5mL) was added dropwise at 0 ℃ to react at room temperature for 1 hour, the reaction solution was concentrated under reduced pressure, the concentrated residue was added to a mixed solvent of DCM (15mL) and methanol (10mL), 7C (1.69g, 5.97mmol) was added thereto, stirring was carried out at room temperature for 30 minutes, sodium triacetoxyborohydride (3.16g, 14.9mmol) was added thereto, and stirring was carried out at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, the mixture was extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and separated by column chromatography (DCM: methanol (v/v) ═ 1: 0-15: 1) to give 7d as a yellow liquid (1.36g, 52% yield).
The fourth step: 2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] -2-fluoro-phenyl ] ethanol (7e)
2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]ethanol
7d (1.3g, 2.5mmol) was dissolved in THF (15mL), tetrabutylammonium fluoride (1.3g, 4.9mmol) was added, and the mixture was stirred at room temperature for 1 hour. Water (50mL) was added, extraction was performed with DCM (50 mL. times.2), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 7e as a yellow liquid (0.89g, 87% yield).
The fourth step: 3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] -2-fluorophenyl ] ethoxy ] acrylic acid tert-butyl ester (intermediate 7)
tert-butyl 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]-2-fluoro-phenyl]ethoxy]propanoate
7e (0.89g, 2.2mmol) was added to acetonitrile (20mL), tert-butyl acrylate (0.83g, 6.5mmol) and benzyltrimethylammonium hydroxide (0.27g, 0.65mmol) were added, and the mixture was stirred at 40 ℃ for 3 hours. Water (50mL) was added and extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulphate, concentrated under reduced pressure and isolated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give intermediate 7 as a yellow liquid (0.94g, 81% yield).
Intermediate 8: 4-pyrazinyl-2-yl-1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-pyrazin-2-yl-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000401
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (0.824g, 4.0mmol) and 2-iodopyrazine (8a) (1.03g, 4.0mmol) were added to DMF (20mL) and stirred at 110 ℃ for 8 h. The reaction mixture was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, followed by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 8(0.4g, yield 30%) as a yellow liquid.
LCMS m/z＝335.2.[M+1]。
Intermediate 9: 4-Pyrimidin-2-yl-1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-pyrimidin-2-yl-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000402
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.03, 4.0mmol) was dissolved in DMF (20mL), and 2-chloropyrimidine (9a) (0.46g, 1.0mmol) and potassium carbonate (1.11g, 8.0mmol) were added in this order and stirred at 50 ℃ for 4 hours. Water (100mL) was added, and extraction was performed with ethyl acetate (200mL, 100mL), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v): 1) to give intermediate 9(1.1.34g, yield 100%) as a yellow liquid.
LCMS m/z＝335.2[M+1]。
Intermediate 10: 4-Thiazol-2-yl-1-oxa-4, 9-diazaspiro [5.5] -undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-thiazol-2-yl-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000403
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.03g, 4.0mmol) was dissolved in DMF (20mL), and 2-bromo-thiazole (10a) (0.655g, 4.0mmol) and cesium carbonate (2.61g, 8.0mmol) were added in that order and stirred at 50 ℃ for 4 hours. Water (100mL) was added, and the mixture was extracted with ethyl acetate (200mL, 100mL), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v): 1) to give intermediate 10(0.28g, yield 21%) as a yellow liquid.
LCMS m/z＝340.1[M+1]。
Intermediate 11: 4- (3-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(3-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000411
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (2.56g, 10.0mmol) was dissolved in 1, 4-dioxane (50mL), 3-chloropyridine (11a) (1.70g, 15.0mmol), sodium tert-butoxide (2.4g, 25.0mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.8g, 1.4mmol), palladium acetate (0.16g, 0.7mmol) were added, and the mixture was refluxed under nitrogen for 8 hours. The reaction mixture was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, followed by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 11(1.1g, yield 33%) as a yellow liquid.
Intermediate 12: 4- (4-Nitro-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-nitro-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000412
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.28g, 4.99mmol) was dissolved in acetonitrile (5mL), and 2-chloro-4-nitropyridine (12a) (0.87g, 5.49mmol) and triethylamine (1.52g, 15.0mmol) were added to the solution, followed by reaction at 140 ℃ for 2 hours in a microwave reactor. The reaction was cooled to room temperature, concentrated under reduced pressure and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 12(0.81g, 43% yield) as a yellow liquid.
Intermediate 13: 4- (4-fluoro-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-fluoro-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000413
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.28g, 4.99mmol) was dissolved in 1, 4-dioxane (20mL), and 2-chloro-4-fluoropyridine (13a) (0.72g, 5.5mmol), sodium tert-butoxide (1.2g, 12.5mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.41g, 0.7mmol), palladium acetate (0.08g, 0.35mmol) were added, and after refluxing under nitrogen for 8 hours. The reaction mixture was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, followed by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 13(0.68g, yield 39%) as a yellow liquid.
Intermediate 14: 4- (3-Nitro-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(3-nitro-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000421
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.28g, 4.99mmol) was dissolved in DMF (20mL), and 2-chloro-3-nitropyridine (14a) (1.19g, 7.5mmol), potassium carbonate (1.38g, 10.0mmol) were added and reacted at 80 ℃ for 8 hours. The reaction was cooled to room temperature, concentrated under reduced pressure and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 14(0.95g, 50% yield) as a yellow liquid.
Intermediate 15: 4- (4-cyclopropyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-cyclopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000422
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.20g, 4.68mmol) was dissolved in 1, 4-dioxane (80mL), and 2-chloro-4-cyclopropylpyridine (15a) (0.79g, 5.15mmol), sodium tert-butoxide (1.12g, 11.7mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.38g, 0.65mmol), palladium acetate (0.07g, 0.33mmol), and refluxed under nitrogen for 8 hours. The reaction mixture was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, followed by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 15(0.95g, yield 54%) as a yellow liquid.
LCMS m/z＝374.2[M+1].
Intermediate 16: 4- [4- (trifluoromethyl) -2-pyridinyl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-[4-(trifluoromethyl)-2-pyridyl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000423
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (5.3g, 21mmol) was dissolved in 1, 4-dioxane (50mL), and 2-chloro-4- (trifluoromethyl) pyridine (16a) (4.1g, 23mmol), sodium tert-butoxide (5.0g, 52.0mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (1.7g, 2.9mmol), palladium acetate (0.3g, 1.4mmol), under nitrogen, refluxed for 8 hours, were added. The reaction mixture was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to obtain intermediate 16(5.4g, yield 65%) as a yellow liquid.
LCMS m/z＝402.2[M+1].
Intermediate 17: 4- [4- (2-Thiophen) pyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-[4-(2-thienyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000431
The first step is as follows: 2-chloro-4- (2-thiophene) pyrimidine (17b)
2-chloro-4-(2-thienyl)pyrimidine
2, 4-dichloropyrimidine (3.0g, 20.1mmol), 2-thiophene-boronic acid (17a) (3.1g, 24.2mmol) were dissolved in a mixed solvent of 1, 4-dioxane (50m1) and water (10ml), and sodium carbonate (4.26g, 40.2mmol), [1, 1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (0.73g, 1.0mmol) was added to react at 90 ℃ for 5 hours under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, and a saturated aqueous solution of sodium hydrogencarbonate (50mL) was added to quench the reaction, which was extracted with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v): 5: 1) to obtain 17b (2.6g, yield 63.4%) as a brown liquid.
LCMS m/z＝197.6[M+1]。
The second step is that: 4- [4- (2-Thiophen) pyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl-4-[4-(2-thienyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
17b (2.6g, 12.75mmol), tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (3.42g, 13.38mmol) was added to DMF (50mL), potassium carbonate (5.3g, 38.25mmol) was added, and the reaction was carried out at 75 ℃ for 5 hours. The reaction mixture was cooled to room temperature, and a saturated aqueous solution of sodium hydrogencarbonate (50mL) was added thereto, followed by extraction with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5: 1) to give intermediate 17(5.0g, yield 92.6%) as a brown liquid.
Intermediate 18: 4- [ 4-Phenoxypyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl-4-(4-phenoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000432
The first step is as follows: 2-chloro-4-phenoxy-pyrimidine (18b)
2-chloro-4-phenoxy-pyrimidine
2, 4-dichloropyrimidine (18a) (5.0g, 33.56mmol) and phenol (3.15g, 33.56mmol) were dissolved in DMF (50ml), and potassium carbonate (6.9g, 50.3mmol) was added to react at 90 ℃ for 5 hours. The reaction mixture was cooled to room temperature, an aqueous solution (100mL) was added thereto, the mixture was extracted with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5: 1) to give 18b (5.0g, yield 72.5%) as a brown liquid.
LCMS m/z＝207.6[M+1]。
The second step is that: 4- [ 4-Phenoxypyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (intermediate 18)
tert-butyl-4-(4-phenoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
2b (1.0g, 4.8mmol) and tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.29g, 5.04mmol) were added to DMF (25mL), potassium carbonate (2g, 14.4mmol) was added, and the reaction was carried out at 75 ℃ for 5 hours. The reaction mixture was cooled to room temperature, and a saturated aqueous solution of sodium hydrogencarbonate (50mL) was added thereto, followed by extraction with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5: 1) to give intermediate 18(1.5g, yield 72.8%) as a brown liquid.
LCMS m/z＝427.5[M+1]。
Intermediate 19: 4- [ 4-Anilinopyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-anilinopyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000441
The first step is as follows: 2-chloro-4-anilino-pyrimidines 19b
2-chloro-N-phenyl-pyrimidin-4-amine
2, 4-dichloropyrimidine 18a (5.0g, 33.56mmol) and aniline (3.15g, 33.56mmol) were dissolved in ethanol (50ml), and triethylamine (5.1g, 50.3mmol) was added to the solution to react at 90 ℃ for 5 hours. The reaction mixture was concentrated under reduced pressure, an aqueous solution (100mL) was added thereto, the mixture was extracted with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, followed by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5: 1) to give 19b (4.5g, yield 65.3%) as a brown liquid.
LCMS m/z＝206.6[M+1]。
The second step is that: 4- [ 4-Anilinopyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (intermediate 19)
tert-butyl 4-(4-anilinopyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
2-chloro-4-anilinopyrimidine (3b) (1.0g, 4.8mmol) and tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.29g, 5.04mmol) were added to DMF (25mL), potassium carbonate (2g, 14.4mmol) was added, and the reaction was carried out at 75 ℃ for 5 hours. The reaction mixture was cooled to room temperature, and a saturated aqueous solution of sodium hydrogencarbonate (50mL) was added thereto, followed by extraction with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5: 1) to give intermediate 19(1.4g, yield 67.9%) as a brown liquid.
LCMS m/z＝426.5[M+1]。
Intermediate 20: 4- [ 5-Methylpyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(5-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000451
2-chloro-5-methylpyrimidine (20b) (1.0g, 7.8mmol) and tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (2.1g, 8.2mmol) were added to DMF (25mL), potassium carbonate (3.2g, 23.4mmol) was added and reacted at 90 ℃ for 5 hours. Aqueous solution (100mL) was added, extraction was performed with ethyl acetate (50mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (petroleum ether/ethyl acetate (v/v): 5: 1) to obtain intermediate 20(2.0g, yield 74.0%) as a brown liquid.
LCMS m/z＝349.4[M+1]。
Intermediate 21: 4- [ 4-Isopropoxypyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-isopropoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000452
2-chloro-4-isopropoxypyrimidine (5a) (0.5g, 2.9mmol) and tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (0.77g, 3mmol) were added to DMF (25mL), potassium carbonate (1.2g, 8.7mmol) was added, and the reaction was carried out at 90 ℃ for 5 hours. Aqueous solution (100mL) was added, extraction was performed with ethyl acetate (50mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5: 1) to give intermediate 21(0.9g, yield 78.9%) as a brown liquid.
LCMS m/z＝393.2[M+1]。
Intermediate 22: 4- (4-isopropyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000453
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (2.56g, 10.0mmol) was added to 1, 4-dioxane (50mL), 2-chloro-4-isopropylpyridine 22a (1.87g, 12.0mmol), sodium tert-butoxide (2.4g, 25.0mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.81g, 1.4mmol) and palladium acetate (0.16g, 0.7mmol) were added, and the mixture was refluxed under nitrogen for 8 hours. Water (100mL) was added, and the mixture was extracted with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v): 1) to obtain intermediate 22(2.5g, yield 67%) as a yellow liquid.
Intermediate 23: 4- [4- (1-hydroxy-1-methylethyl) pyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-[4-(1-hydroxy-1-methyl-ethyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000461
The first step is as follows: 2- (2-Chloropyrimidin-4-yl) propan-2-ol (23b)
2-(2-chloropyrimidin-4-yl)propan-2-ol
Methyl 2-chloropyrimidine-4-carboxylate (23a) (1.00g, 5.79mmol) was added to THF (30mL) under nitrogen, a solution of methylmagnesium bromide in THF (6mL, 3mol/L) was slowly added at-78 deg.C and the reaction was allowed to proceed at room temperature for 1.5 hours. Water (100mL) was added, extraction was performed with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give 23b as a brown liquid (0.63g, 63% yield).
The second step is that: 4- [4- (1-hydroxy-1-methylethyl) pyrimidin-2-yl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (intermediate 23)
tert-butyl 4-[4-(1-hydroxy-1-methyl-ethyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
23b (0.95g, 3.7mmol) was added to DMF (50mL), and 23b (0.61g, 3.5mmol), potassium carbonate (0.98g, 7.1mmol) were added and reacted at 75 ℃ under nitrogen for 5 hours. Water (100mL) was added, extraction was performed with DCM (100 mL. times.2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give intermediate 23 as a yellow liquid (1.01g, 73% yield).
Intermediate 24: 4- (4-Phenylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-phenylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000462
The first step is as follows: 2-chloro-4-phenylpyrimidine (24b)
2-chloro-4-phenyl-pyrimidine
24a (1.5g, 10.0mmol) was added to a mixed solvent of 1, 4-dioxane (52mL) and water (13mL), and phenylboronic acid (1.8g, 15.0mmol), sodium carbonate (2.7g, 25.0mmol), PdCl2(dppf) (cas: 72287-26-4) (0.37g, 0.5mmol) were added. The reaction was carried out at 90 ℃ for 5 hours. The reaction mixture was filtered through celite, water (100mL) was added to the filtrate, and extraction was performed with ethyl acetate (50mL × 3), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give 24b (1.32g, yield 70%) as a white solid.
The second step is that: 4- (4-Phenylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (intermediate 24)
tert-butyl 4-(4-phenylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.86g, 7.3mmol) was added to DMF (50mL), 24b (1.32g, 6.9mmol), potassium carbonate (1.91g, 13.8mmol) were added, and the reaction was carried out under nitrogen at 75 ℃ for 5 hours. Water (100mL) was added, and the mixture was extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give intermediate 24(2.01g, yield 71%) as a yellow liquid.
Intermediate 25: 4- (4-methoxyphenyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-methoxyphenyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000471
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.28g, 5.0mmol) was added to 1, 4-dioxane (50mL), 1-bromo-4-methoxybenzene 25a (1.4g, 7.5mmol), sodium tert-butoxide (1.2g, 12.5mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.4g, 0.7mmol), palladium acetate (0.08g, 0.35mmol), under nitrogen, refluxed for 8 hours. Water (100mL) was added, and extraction was performed with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v): 1) to obtain intermediate 25(0.96g, yield 53%) as a yellow liquid.
Intermediate 26: 4- (1-methylpyrazol-4-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(1-methylpyrazol-4-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000472
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.28g, 5.0mmol) was added to 1, 4-dioxane (50mL), 4-bromo-1-methylpyrazole (1.21g, 7.49mmol), sodium tert-butoxide (1.2g, 12.5mmol), tri-tert-butylphosphine (0.1g, 0.5mmol), palladium acetate (0.08g, 0.35mmol) were added, and the mixture was refluxed under nitrogen for 8 hours. Water (100mL) was added, and extraction was performed with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 4: 1) to give intermediate 26(1.0g, yield 50%) as a yellow liquid.
Intermediate 27: 4- (3-cyano-2-pyridinyl) -1-oxo-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(3-cyano-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000473
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.28g, 5.0mmol) was added to THF (50mL), 2-fluoropyridine-3-carbonitrile (0.92g, 7.5mmol), triethylamine (1.52g, 15.0mmol) were added, and the mixture was refluxed at 80 ℃ for 8 hours. The reaction mixture was cooled to room temperature, concentrated under reduced pressure and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 27(1.7g, yield 95%) as a yellow liquid.
Intermediate 28: 4- (2-chlorophenyl) -1-oxo-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(2-chlorophenyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000481
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.28g, 5.0mmol) was added to 1, 4-dioxane (50mL), 1-bromo-2-chlorobenzene (1.43g, 7.5mmol), sodium tert-butoxide (1.2g, 12.5mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.40g, 0.7mmol), palladium acetate (0.08g, 0.35mmol) were added, and the reaction was carried out at 100 ℃ under nitrogen for 8 hours. Water (100mL) was added, and extraction was performed with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 28(1.2g, yield 65%) as a yellow liquid.
Intermediate 29: [2- (1-oxa-4, 9-diaza [5.5] undecan-4-yl) -4-pyridinyl ] methanol
[2-(1-oxa-4，9-diazaspiro[5.5]undecan-4-yl)-4-pyridyl]methanol
Figure GPA0000261095610000482
The first step is as follows: 4- [4- [ [ tert-butyl (dimethyl) silyl ] oxymethyl ] -2-pyridyl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (29b)
tert-butyl 4-[4-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-pyridyl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.64g, 6.4mmol) was added to 1, 4-dioxane (30mL), and 4- (((tert-butylmethylsilyloxy) methyl) -2-chloropyridine (29a) (prepared with reference to WO2008074752A 2) (1.65g, 6.4mmol), sodium tert-butoxide (1.54g, 16.0mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.26g, 0.45mmol) and palladium acetate (0.072g, 0.32mmol) were added and refluxed under nitrogen for 8 hours. After the reaction was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 29b (2.0g, yield 65%) as a yellow solid.
LCMS m/z＝478.2[M+1].
The second step is that: [2- (1-oxa-4, 9-diaza [5.5] undecan-4-yl) -4-pyridinyl ] methanol (intermediate 29)
[2-(1-oxa-4，9-diazaspiro[5.5]undecan-4-yl)-4-pyridyl]methanol
29b (1.7g, 3.56mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (3mL) was added and stirred at room temperature for 2 h. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain trifluoroacetate salt of intermediate 29 (1.34, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝264.3[M+1].
Intermediate 30: 4- (2-chloro-4-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(2-chloro-4-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000491
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (2.56g, 10.0mmol) was added to 1, 4-dioxane (50mL), and 2-chloro-4-bromo-pyridine (30a) (2.1g, 11.0mmol), sodium tert-butoxide (2.4g, 25.0mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.58g, 1.0mmol) and palladium acetate (0.11g, 0.5mmol) were added in this order, and after replacement with nitrogen, a nitrogen atmosphere was refluxed for 8 hours. After the reaction was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 30(2.9g, yield 80%) as a yellow solid.
LCMS m/z＝368.2[M+1]。
Intermediate 31: 4- (4-methoxy-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-methoxy-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000492
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (5.12g, 20.0mmol) was added to 1, 4-dioxane (60mL), 2-chloro-4-methoxy-pyridine (4.3g, 30.0mmol), cesium carbonate (13.0g, 40.0mmol), 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene (1.6g, 2.8mmol) and palladium acetate (0.3g, 1.4mmol) were added, and the mixture was refluxed under nitrogen for 8 hours. After the reaction was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether (v/v) ═ 1: 2) to give intermediate 31(6.5g, yield 90%) as a yellow oil.
Intermediate 32: 4-phenyl-1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-phenyl-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000493
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (5.12g, 20.0mmol) was added to 1, 4-dioxane (50mL), bromobenzene (4.7g, 30.0mmol), sodium tert-butoxide (3.84g, 40.0mmol), 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene (1.6g, 2.8mmol) and palladium acetate (0.3g, 1.4mmol) were added, and the mixture was refluxed under nitrogen for 8 hours. After the reaction was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether (v/v) ═ 1: 10) to give intermediate 32(4.6g, yield 69%) as a yellow oil.
Intermediate 33: 4- (4-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000501
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (5.12g, 20.0mmol) was added to 1, 4-dioxane (50mL), 4-chloro-pyridine (3.4g, 30.0mmol), sodium tert-butoxide (3.84g, 40.0mmol), 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene (1.6g, 2.8mmol) and palladium acetate (0.3g, 1.4mmol) were added, and the mixture was refluxed under nitrogen for 10 hours. After the reaction was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate/petroleum ether (v/v) ═ 1: 1) to give intermediate 33(3.8g, yield 57%) as a yellow oil.
Intermediate 34: 4- (4-fluoro-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-fluoro-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000502
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.28g, 5.0mmol) was added to 1, 4-dioxane (30mL), 2-chloro-4-fluoro-pyridine (0.72g, 5.5mmol), sodium tert-butoxide (1.2g, 12.5mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.41g, 0.7mmol), palladium acetate (0.079g, 0.35mmol), under nitrogen, refluxed for 8 hours. After the reaction was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 34(0.68g, yield 39%) as a yellow solid.
LCMS m/z＝352.3[M+1].
Intermediate 35: 4- (4-isopropylphenyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(4-isopropylphenyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000503
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.28g, 5.0mmol) was added to 1, 4-dioxane (30mL), 1-chloro-4-isopropylbenzene (35a) (1.1g, 5.5mmol), sodium tert-butoxide (1.2g, 12.5mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.41g, 0.7mmol), palladium acetate (0.079g, 0.35mmol) were added, and the mixture was refluxed under nitrogen for 8 hours. After the reaction was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (200mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 35(1.68g, yield 90%) as a yellow solid.
LCMS m/z＝375.3[M+1].
Intermediate 36: 4- (3-Phenylmethyloxycarbonyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester
tert-butyl 4-(3-benzyloxycarbonyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Figure GPA0000261095610000511
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (1.3g, 5.0mmol) was placed in a microwave reaction tube (20mL), and 1, 4-dioxane (10mL), 36a (1.2g, 5.0mmol), N, N-diisopropylethylamine (1.9g, 15.0mmol) was added and microwave reacted at 130 ℃ for 2 hours. After the reaction was cooled to room temperature, the reaction solution was concentrated under reduced pressure and separated by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give intermediate 36(1.8g, 77% yield) as a yellow solid.
LCMS m/z＝468.3[M+1]。
The Boc removing reaction conditions of the intermediates 8 to 28 and the intermediates 30 to 36 are as follows: dissolving a substrate in DCM, adding trifluoroacetic acid (1/5-1/2 in volume of DCM), and reacting at room temperature until the reaction is finished. The reaction liquid is directly decompressed and concentrated to obtain the corresponding trifluoroacetic acid salt of the de-Boc product.
Example 1: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-heteropiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 1)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000512
The first step is as follows: 1- (Cyclopropylmethyl) pyrazole-3-carboxylic acid ethyl ester (1B)
ethyl 1-(cyclopropylmethyl)pyrazole-3-carboxylate
Sodium hydride (5.7g, 143.7mmol, 60% w/w) was weighed out and added to DMF (100mL), a solution of 1A (10.0g, 71.4mmol) in DMF (100mL) was added dropwise at 0 ℃ and after completion of the addition, stirred at 0 ℃ for 30 minutes, bromomethylcyclopropane (14.5g, 107.0mmol) was added dropwise and stirred at room temperature for 3 hours. The reaction was quenched by dropwise addition of water (150mL), extracted with ethyl acetate (150mL × 2), and the combined organic phases were washed with saturated brine (100mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2: 1) to give 1B as a yellow liquid (6.5g, yield 47%).
1H NMR(400MHz，CDCl3)δ7.52-7.51(d，1H)，6.80-6.79(d，1H)，4.40-4.35(m，2H)，4.05-4.04(d，2H)，1.37-1.32(m，3H)，1.26-1.23(m，1H)，0.64-0.62(m，2H)，0.37-0.35(m，2H).
The second step is that: 1- (cyclopropylmethyl) pyrazole-3-carboxylic acid (1C)
1-(cyclopropylmethyl)pyrazole-3-carboxylic acid
THF (60mL), methanol (15mL) and water (15mL) were added to 1B (3.86g, 20.0mmol), stirred well, added lithium hydroxide (0.95g, 40.0mmol) and stirred at room temperature for 3 hours. The reaction was directly concentrated under reduced pressure to remove most of THF and methanol, water (100mL) was added, the pH of the aqueous phase was adjusted to 6 with dilute hydrochloric acid (1.0M), ethyl acetate was extracted (150 mL. times.2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 1C as a white solid (3.2g, 100% yield).
LCMS m/z＝167.1[M+1]。
The third step: 4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1D)
tert-butyl 4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1C (3.2g, 20.0mmol) and tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (5.3g, 20.0mmol) were added to DCM (50mL), triethylamine (13.0mL, 100.0mmol) and 1-propylphosphoric anhydride (17.0g, 30.0mmol, 50% ethyl acetate solution) were added, and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extraction was performed, the aqueous phase was extracted with DCM (100mL × 1), the combined organic phases were dried over anhydrous sodium sulfate, filtration was performed, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 1D as a yellow solid (2.4g, yield 33%).
1H NMR(400MHz，CDCl3)δ7.45-7.44(d，1H)，6.69-6.68(d，1H)，4.69-3.94(m，4H)，3.75-3.55(m，7H)，3.22-3.09(m，2H)，1.80-1.76(m，2H)，1.56-1.54(m，1H)，1.42(s，9H)，1.25-1.20(m，1H)，0.63-0.59(m，2H)，0.35-0.32(m，2H).
LCMS m/z＝427.2[M+23]。
The fourth step: [1- (cyclopropylmethyl) pyrazol-3-yl ] - (1-oxa-4, 9-diaza [5.5] undecan-4-yl) methanone (1E)
[1-(cyclopropylmethyl)pyrazol-3-yl]-(1-oxa-4，9-diazaspiro[5.5]undecan-4-yl)methanone
1D (2.4g, 5.9mmol) was dissolved in DCM (30mL), and trifluoroacetic acid (6mL) was added and stirred at room temperature for 2 hours. The reaction mixture was directly concentrated under reduced pressure to remove the solvent, and toluene (5mL) was added to the residue to obtain 1E trifluoroacetate salt (2.5g, 92% yield) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝305.2[M+23]。
The fifth step: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (1F)
tert-butyl 3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 1E (1.2g, 3.0mmol) was placed in acetonitrile (25mL), intermediate 1(1.1g, 3.0mmol), potassium carbonate (2.0g, 15.0mmol) and water (0.5mL) were added and stirred at 60 ℃ for 24 h. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to obtain 1F as a yellow liquid (1.64g, yield 96%).
1H NMR(400MHz，CDCl3)δ7.47-7.46(m，1H)，7.18-7.16(m，1H)，7.04-7.01(m，3H)，6.72-6.71(m，1H)，4.13-3.97(m，3H)，3.75-3.65(m，9H)，2.87-2.45(m，13H)，2.03-1.58(m，4H)，1.48(s，9H)，0.68-0.62(m，2H)，0.38-0.34(m，2H).
LCMS m/z＝581.4[M+1]。
And a sixth step: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (1G)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
1F (1.64g, 2.82mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction mixture was directly concentrated under reduced pressure to remove the reaction solvent, and toluene (10mL) was added to the residue, followed by concentration under reduced pressure to obtain 1G trifluoroacetate salt (1.48G, yield 100%) as a yellow liquid.
LCMS m/z＝525.4[M+1]。
The seventh step: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (1H)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-methyl-propanamide
1G of trifluoroacetate salt (1.5G, 2.8mmol) was dissolved in DCM (20mL), methylaminoacetaldehyde dimethyl acetal (0.5G, 4.2mmol), triethylamine (1.43G, 14.1mmol) and HATU (1.61G, 4.2mmol) were added, and the mixture was stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 1H as a yellow liquid (1.3g, yield 73.6%).
LCMS m/z＝626.5[M+1]。
Eighth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (1I)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
1H (0.63g, 1.0mmol) was dissolved in THF (10mL), and TsOH. H.was added2O (0.96g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.64g, 3.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 1I as a yellow solid (0.50g, yield 55%).
LCMS m/z＝449.8[M/2+1]。
The ninth step: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-heteropiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 1)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
1I (0.50g, 0.56mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.18g, 1.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL) in that order, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was separated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give compound 1(0.35g, yield 80%) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.08(br，1H)，7.47(s，1H)，7.12-6.93(m，5H)，6.69-6.67(m，2H)，6.36(br，2H)，5.48(br，1H)，4.08-3.95(m，4H)，3.62-3.47(m，10H)，3.05-2.62(m，18H)，1.93-1.84(m，5H)，1.28-1.25(m，2H)，0.63-0.61(m，2H)，0.35-0.33(m，2H).
LCMS m/z＝784.5[M+1]。
Example 2: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 2)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000541
The first step is as follows: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl-3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (2A)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
1H (0.63g, 1.0mmol) was dissolved in THF (10mL), and TsOH. H.was added 2O (0.96g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to obtain a crude product, dissolving the crude product in a mixed solvent of DCM (10mL) and methanol (3mL), adding 8- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silicon dioxideBase of]Oxy-ethyl radical]-5-hydroxy-4H-1, 4-benzoxazin-3-one (0.34g, 1.0mmol) was stirred at room temperature for 30 minutes, then sodium triacetoxyborohydride (0.64g, 3.0mmol) was added, and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 2A as a yellow solid (0.450g, 50% yield).
LCMS m/z＝451.9[M/2+1]。
The second step is that: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 2)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-91-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
2A (0.45g, 0.50mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.16g, 1.0mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 2(0.28g, 71% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ7.48(s，1H)，7.16-7.12(m，1H)，6.97-6.87(m，4H)，6.53-6.51(m，1H)，6.51-6.45(m，1H)，5.00-4.94(m，1H)，4.48-4.42(m，2H)，4.16-3.97(m，4H)，3.65-3.42(m，10H)，2.94-2.54(m，20H)，1.95-1.77(m，4H)，1.28-1.25(m，2H)，0.63-0.61(m，2H)，0.36-0.33(m，2H).
LCMS m/z＝788.5[M+1]。
Example 3: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 3)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000551
Figure GPA0000261095610000561
The first step is as follows: 4- (2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (3B)
tert-butyl 4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (5.12g, 20.0mmol) was added to 1, 4-dioxane (50mL), 2-chloropyridine (3A) (3.4g, 30.0mmol), cesium carbonate (13.0g, 40.0mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (1.6g, 2.8mmol) and palladium acetate (0.3g, 1.4mmol) were added, and the mixture was refluxed under nitrogen for 8 hours. The reaction mixture was cooled to room temperature, water (100mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (100mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 3B as a yellow liquid (3.0g, yield 45%).
LCMS m/z＝334.3[M+1]。
The second step is that: 4- (2-pyridinyl) -1-oxa-4, 9-diaza [5.5] undecane (3C)
4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane
3B (0.5g, 1.5mmol) was dissolved in DCM (20 mL). Trifluoroacetic acid (4mL) was added to the reaction flask, and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain a yellow liquid of 3C trifluoroacetate salt (0.53g, yield 100%).
The third step: 3- [2- [3- [ [4- (2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (3D)
tert-butyl 3-[2-[3-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
After 3C trifluoroacetate salt (0.53g, 1.5mmol) was placed in acetonitrile (15mL), intermediate 2(0.53g, 1.5mmol), potassium carbonate (1.1g, 7.5mmol) and water (0.5mL) were added in this order, and the mixture was stirred at room temperature for 8 hours. Water (50mL) was added, and extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 3D as a yellow solid (0.50g, yield 65%).
LCMS m/z＝496.4[M+1]。
The fourth step: 3- [2- [3- [ [4- (2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (3E)
3-[2-[3-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
3D (0.50g, 1.0mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain the trifluoroacetate salt of 3E (0.45g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝440.4[M+1]。
The fifth step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (3F)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 3E (0.45g, 1.0mmol) was dissolved in DCM (20mL), triethylamine (0.52g, 5.0mmol), methylaminoacetaldehyde dimethyl acetal (0.18g, 1.5mmol), HATU (0.58g, 1.5mmol) were added, and the mixture was stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, the combined organic phases were extracted with DCM (100mL × 2), washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 3F as a yellow solid (0.50g, yield 90.0%).
LCMS m/z＝541.4[M+1]。
And a sixth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-N-methyl-3- [2- [3- [ [4- (2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (3G)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
3F (0.50g, 0.92mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.88g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.31g, 0.92mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.59g, 2.8mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 3G (0.33G, yield 44%) as a yellow solid
LCMS m/z＝407.4[M/2+1]。
The seventh step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 3)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
3G (0.33G, 0.41mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.65G, 4.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 3(0.20g, 71% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.32-8.29(d，1H)，7.93(br，1H)，7.76-7.74(m，1H)，7.28-7.14(m，4H)，7.07-7.05(d，1H)，6.94-6.92(d，1H)，6.81-6.78(m，1H)，6.57-6.55(d，1H)，5.30-5.27(m，1H)，4.20(s，2H)，3.69-3.67(m，2H)，3.62-3.38(m，9H)，3.22-3.03(m，13H)，2.78-2.75(m，2H)，2.57-2.56(m，2H)，2.13-2.09(m，2H)，1.75-1.72(m，2H).
LCMS m/z＝699.5[M+1]。
Example 4: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [4- [2- [4- (2-pyridyl) -1-oxa-4, 9-heterospirodiaza [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -propionamide (Compound 4)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000581
The first step is as follows: 3- [2- [4- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (4A)
tert-butyl 3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 3C (0.5g, 1.5mmol) was added to acetonitrile (15mL), intermediate 3(0.55g, 1.5mmol), potassium carbonate (1.1g, 7.5mmol) and water (0.5mL) were added, and the mixture was stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 4A as a yellow liquid (0.50g, yield 64%).
LCMS m/z＝510.5[M+1]。
The second step is that: 3- [2- [4- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (4B)
3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
4A (0.90g, 1.8mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain the trifluoroacetic acid salt of 4B (0.80g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝454.3[M+1]。
The third step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [4- [2- [4- (2-pyridyl) -1-oxa-4, 9-diaza [5.5] undec-3-yl ] ethyl ] phenyl ] ethoxy ] propanamide (4C)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 4B (0.8g, 1.8mmol) was dissolved in DCM (20mL), and triethylamine (0.89g, 8.8mmol), methylaminoacetaldehyde dimethyl acetal (0.32g, 2.6mmol) and HATU (1.0g, 2.6mmol) were added successively and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, and extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 4C as a yellow liquid (0.55g, 56% yield).
LCMS m/z＝555.3[M+1]。
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [4- [2- [4- (2-pyridyl) -1-oxa-4, 9-heterospirodiaza [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (4D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
4C (0.55g, 0.99mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.94g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.63g, 3.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 4D as a yellow solid (0.34g, yield 41%).
The fifth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [4- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 4)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
4D (0.34g, 0.41mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (1.98g, 12.3mmol) was added, and the mixture was stirred at room temperature for 12 hours. To the reaction solution, saturated aqueous sodium bicarbonate (50mL) was sequentially added dropwise, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 4(0.12g, 41% yield) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.22(br，1H)8.16-8.19(d，1H)，8.08-8.10(dd，1H)，7.49-7.53(m，1H)，7.05-7.11(m，5H)，6.90-6.92(d，1H)，6.81-6.83(d，1H)，6.60-6.63(dd，1H)，5.74(s，1H)，6.47-6.49(dd，1H)，4.98-5.02(m，1H)，3.69-3.71(m，2H)，3.59-3.62(m，2H)，3.52-3.57(m，2H)，3.43-3.45(m，2H)，3.37(s，2H)，3.29-3.34(m，4H)，2.92(s，2H)2.65-2.77(m，10H)，2.32-2.37(m，3H)，1.73-1.77(m，2H)1.52-1.57(m，2H)1.15-1.28(br，2H)。
LCMS m/z＝713.5[M+1]。
Example 5: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [4- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (Compound 5)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000591
Figure GPA0000261095610000601
The first step is as follows: 2-Propynylthiazole-4-carboxylic acid ethyl ester (5B)
ethyl 2-prop-1-ynylthiazole-4-carboxylate
Ethyl 2-bromothiazole-4-carboxylate (5A) (11.8g, 50.0mmol) was dissolved in THF (200mL), and triethylamine (7.6g, 75.0mmol), triphenylphosphine (0.33g, 1.25mmol), ditriphenylphosphine palladium dichloride (1.75g, 2.5mmol), propyne (120g, 100.0mmol, 3-4% heptane solution) were added in this order and stirred at 60 ℃ for 4 hours. Most of the reaction solvent was removed under reduced pressure, water (150mL) was added, extraction was performed with ethyl acetate (200mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was chromatographed on a silica gel column (petroleum ether/ethyl acetate (v/v) ═ 10: 1) to give 5B as a yellow solid (4.8g, yield 49.2%).
1H NMR(400MHz，CDCl3)δ8.06(s，1H)，4.41-4.36(q，2H)，2.07(s，3H)，1.39-1.35(t，3H).
The second step is that: 2-Propynylthiazole-4-carboxylic acid (5C)
2-prop-1-ynylthiazole-4-carboxylic acid
5B (4.8g, 24.6mmol) was dissolved in THF (100mL), methanol (10mL), water (20mL) and lithium hydroxide (1.8g, 73.8mmol) were added, and the mixture was stirred at room temperature for 2 hours. Most of THF and methanol were removed under reduced pressure, 1.0M dilute aqueous hydrochloric acid was added dropwise to adjust the pH to 6, extraction was performed with ethyl acetate (150 mL. times.2), the combined organic phases were dried over anhydrous sodium sulfate, and concentration under reduced pressure gave 2-propynylthiazole-4-carboxylic acid (5C) as a white solid (4.1g, yield 100%).
The third step: 4- (2-Propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (5D)
tert-butyl 4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
5C (4.2g, 25.1mmol) and tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (6.4g, 25.1mmol) were added to DCM (50mL), triethylamine (17.5mL, 126.0mmol), 1-propylphosphoric anhydride (24.0g, 37.7mmol, 50% ethyl acetate solution) were added, and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, the layers were extracted, the aqueous phase was extracted with DCM (200mL × 1), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 5D as a yellow liquid (4.2g, yield 41.2%).
The fourth step: 1-oxa-4, 9-diaza [5.5] undecan-4-yl- (2-propynylthiazol-4-yl) methanone (5E)
1-oxa-4，9-diazaspiro[5.5]undecan-4-yl-(2-prop-1-ynylthiazol-4-yl)methanone
5D (1.4g, 3.5mmol) was weighed out and dissolved in DCM (20mL), and trifluoroacetic acid (4mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain 5E trifluoroacetate salt (1.5g, yield 100%) as a yellow liquid after concentration under reduced pressure.
The fifth step: 3- [2- [4- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (5F)
tert-butyl 3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 5E (1.05g, 2.9mmol) was placed in acetonitrile (15mL), intermediate 4(1.0g, 2.9mmol), potassium carbonate (2.0g, 15.0mmol) and water (0.5mL) were added and the mixture was stirred at room temperature for 4 hours. Water (50mL) was added, and extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 5F as a yellow solid (1.0g, yield 60%).
LCMS m/z＝568.3[M+1]。
And a sixth step: 3- [2- [4- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (5G)
3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
5F (1.0g, 1.8mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 5G trifluoroacetate salt (0.9G, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝512.3[M+1]。
The seventh step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [4- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (5H)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
5G of trifluoroacetate salt (1.0G, 2.0mmol) was dissolved in DCM (20mL), triethylamine (0.99G, 10.0mmol), methylaminoacetaldehyde dimethyl acetal (0.35G, 3.0mmol) and HATU (1.1G, 3.0mmol) were added, and the mixture was stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 5H as a yellow solid (1.0g, yield 83.0%).
LCMS m/z＝613.4[M+1].
Eighth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethan-N-methyl-3- [2- [4- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (5I)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
5H (1.0g, 1.6mmol) was dissolved in THF (10mL), and TsOH. H.was added2O (1.6g, 8.2mmol), stirring at 40 deg.C for 1 hour, adding saturated aqueous sodium bicarbonate solution (50mL) dropwise, extracting with DCM (100 m.times.2), drying the combined organic phases over anhydrous sodium sulfate, and concentrating under reduced pressure to give a crude product which is dissolved in DCM (10mL) and methanol (3mL)Adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.55g, 1.6mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (1.0g, 4.9mmol), and stirred at room temperature for 3 hours. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (50mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 5I as a yellow solid (0.80g, 55% yield).
LCMS m/z＝886.5[M+1]。
The ninth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [4- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (Compound 5)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
5I (0.80g, 0.90mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (1.5g, 10.0mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 5(0.30g, 43% yield) as a yellow solid.
LCMS m/z＝771.5[M+1]。
1H NMR(400MHz，CDCl3)δ7.91-7.88(m，1H)，7.21-7.18(m，2H)，7.09-6.96(m，3H)，6.77-6.73(m，2H)，6.34-6.30(m，1H)，5.35-5.29(m，1H)，3.92-3.65(m，14H)，3.24-2.45(m，15H)，2.11(s，3H)，1.83-1.61(m，4H).
Example 6: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 6)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000621
The first step is as follows: tert-butyl 3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionate (6A)
tert-butyl 3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 5E (0.50g, 1.2mmol) was added to acetonitrile (15mL), and intermediate 1(0.44g, 1.2mmol), potassium carbonate (0.86g, 6.0mmol) and water (0.5mL) were added and stirred at 60 ℃ for 24 hours. Water (50mL) was added, and extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 6A (0.5g, yield 70%) as a yellow solid.
LCMS m/z＝582.4[M+1]。
The second step is that: trifluoroacetate salt of 3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionic acid (6B)
3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
6A (0.5g, 0.86mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (4mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 6B trifluoroacetate salt (0.45g, yield 100%) as a yellow liquid after concentration under reduced pressure.
The third step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanamide (6C)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 6B (0.45g, 0.86mmol) was dissolved in DCM (15mL), and triethylamine (0.43g, 4.3mmol), methylaminoacetaldehyde dimethyl acetal (0.15g, 1.3mmol), and HATU (0.49g, 1.3mmol) were added successively and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 6C as a yellow solid (0.50g, 93.0% yield).
LCMS m/z＝627.4[M+1]。
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (6D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
6C (0.50g, 0.80mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.76g, 4.0mmol), stirring at 40 ℃ for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.27g, 0.80mmol), stirred at room temperature for 30 minutes, added sodium triacetoxyborohydride (0.51g,2.4mmol), stirred at room temperature for 3 hours. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (50mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 6D as a yellow solid (0.16g, yield 22%)
LCMS m/z＝450.4[M/2+1]。
The fifth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 6)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
6D (0.16g, 0.18mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.29g, 1.8mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 6(0.10g, 72% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.08-8.02(m，1H)，7.94-7.92(m，1H)，7.15-7.08(m，2H)，6.98-6.80(m，4H)，6.56-6.48(m，1H)，5.13-5.07(m，1H)，3.95-3.43(m，10H)，2.98-2.36(m，24H)，2.20-1.67(m，4H)。
LCMS m/z＝785.5[M+1]。
Example 7: 3- [2- [3- [2- [4- (3-ethynylbenzoyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 7)
3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000641
The first step is as follows: 4- (3-ethynylbenzoyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (7B)
tert-butyl 4-(3-ethynylbenzoyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
3-ethynylbenzoic acid (7A) (3.0g, 20.0mmol) and tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (5.2g, 20.0mmol) were placed in DCM (100mL) and triethylamine (14.0mL, 100.0mmol), 1-propylphosphoric anhydride (19.0g, 30.0mmol, 50% ethyl acetate solution) were added successively and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (100mL) was added dropwise, the layers were separated by extraction, the aqueous phase was extracted with DCM (200mL × 1), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 7B as a yellow liquid (5.0g, 64% yield).
The second step is that: trifluoroacetate salt of (3-ethynylphenyl) - (1-oxa-4, 9-diaza [5.5] undecan-4-yl) methanone (7C)
(3-ethynylphenyl)-(1-oxa-4，9-diazaspiro[5.5]undecan-4-yl)methanone
7B (0.6g, 1.6mmol) was dissolved in DCM (20mL), and trifluoroacetic acid (4mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain 7C trifluoroacetate salt (0.65g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝285.3[M+1]。
The third step: tert-butyl 3- [2- [3- [2- [4- (3-ethynylbenzoyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionate (7D)
tert-butyl 3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
7C trifluoroacetate (0.65g, 1.6mmol) was added to acetonitrile (15mL), and intermediate 1(0.57g, 1.6mmol), potassium carbonate (1.1g, 7.9mmol) and water (0.5mL) were added and stirred at 60 ℃ for 24 hours. Water (50mL) was added, ethyl acetate (100mL × 2) was added, the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM/methanol (v/v) ═ 15: 1) to give 7D as a yellow solid (0.65g, 73% yield).
LCMS m/z＝561.4[M+1]。
The fourth step: 3- [2- [3- [2- [4- (3-ethynylbenzoyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (7E)
3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
7D (0.65g, 1.2mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (4mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain the trifluoroacetate salt of 7E (0.59g, yield 100%) as a yellow liquid after concentration under reduced pressure.
The fifth step: n- (2, 2-Dimethoxyethyl) -3- [2- [3- [2- [4- (3-ethynylbenzoyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-methyl-propionamide (7F)
N-(2，2-dimethoxyethyl)-3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
The trifluoroacetate salt of 7E (0.59g, 1.2mmol) was dissolved in DCM (15mL), and triethylamine (0.81mL, 6.0mmol), methylaminoacetaldehyde dimethyl acetal (0.21g, 1.8mmol), HATU (0.67g, 1.8mmol) were added successively and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 7F as a yellow solid (0.61g, yield 86.0%).
LCMS m/z＝606.5[M+1]。
And a sixth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (3-ethynylbenzoyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-methyl-propionamide (7G)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
7F (0.61g, 1.0mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.96g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.64g, 3.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 7G as a yellow solid (0.56G, yield 63%).
LCMS m/z＝878.6[M+1]。
The seventh step: 3- [2- [3- [2- [4- (3-ethynylbenzoyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 7)
3-[2-[3-[2-[4-(3-ethynylbenzoyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
7G (0.56G, 0.64mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (1.0G, 6.4mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 7(0.20g, 41% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.02-7.98(m，1H)，7.56-7.48(m，2H)，7.38-7.30(m，2H)，7.16-7.12(m，1H)，7.05-6.79(m，5H)，6.51-6.42(m，1H)，5.13-5.07(m，1H)，3.72-3.43(m，16H)，3.25-2.40(m，18H)，1.89-1.56(m，2H)。
LCMS m/z＝765.5[M+1]。
Example 8: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide (Compound 8)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
Figure GPA0000261095610000661
The first step is as follows: 4- (1, 2-Benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (8B)
tert-butyl 4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (3.1g, 12.1mmol) was added to acetonitrile (20mL), 3-chloro-1, 2-benzisothiazole (8A) (2.3g, 13.3mmol), 1, 5-diazabicyclo [5.4.0] undec-5-ene (DBU, 1.8g, 12.1mmol) were added, and stirring was carried out at 50 ℃ for 24 hours. After concentration under reduced pressure, most of the reaction solvent was removed, and silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 4: 1) gave 8B (2.5g, yield 53%) as a yellow liquid.
1H NMR(400MHz，CDCl3)δ7.66-7.63(m，2H)，7.59-7.55(m，1H)，7.34-7.30(m，1H)，3.79-3.76(m，4H)，3.12-3.02(m，4H)，2.86(s，2H)，1.96-1.93(m，2H)，1.48-1.41(m，11H)。
LCMS m/z＝412.3[M+23]。
The second step is that: 4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane (8C)
4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecane
8B (0.78g, 2.0mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (4mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 8C trifluoroacetate salt (0.84g, yield 100%) as a yellow liquid after concentration under reduced pressure.
The third step: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (8D)
tert-butyl 3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 8C (0.84g, 2.0mmol) was added to acetonitrile (15mL), and intermediate 1(0.71g, 2.0mmol), potassium carbonate (1.38g, 10.0mmol) and water (0.5mL) were added in that order and stirred at 60 ℃ for 24 hours. Water (50mL) was added, ethyl acetate (100mL × 2) was added, the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM/methanol (v/v) ═ 15: 1) to give 8D as a yellow solid (0.85g, yield 75%).
1H NMR(400MHz，CDCl3) δ 7.89-7.86(d, 1H), 7.83-7.81(d, 1H), 7.47-7.43(m, 1H), 7.37-7.33(m, 1H), 7.22-7.18(m, 1H), 7.06-7.05(m, 3H), 3.97-3.94(m, 2H), 3.72-3.68(m, 4H), 3.49-3.46(m, 2H), 3.33(s, 2H), 2.87-2.47(m, 12H), 2.18-2.14(m, 2H), 1.81-1.75(m, 2H), 1.44(s, 9H). The fourth step: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5]]Undec-9-yl]Ethyl radical]Phenyl radical]Ethoxy radical]Propionic acid (8E)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
8D (1.0g, 1.8mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (4mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain the trifluoroacetic acid salt of 8E (0.9g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝510.3[M+1]。
The fifth step: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-butyl- (2, 2-dimethoxyethyl) propanamide (8F)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-(2，2-dimethoxyethyl)propanamide
The trifluoroacetate salt of 8E (0.90g, 1.8mmol) was dissolved in DCM (15mL), and triethylamine (0.89g, 8.8mmol), N- (2, 2-dimethoxyethyl) butyl-1-amine (intermediate 5) (0.31g, 1.9mmol), HATU (1.0g, 2.6mmol) and the mixture were added successively and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 8F as a yellow solid (1.2g, yield 100%).
LCMS m/z＝653.5[M+1]。
And a sixth step: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-butyl-N- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide (8G)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
8F (0.65g, 1.0mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.95g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.63g, 3.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 8G (0.45G, 49% yield) as a yellow solid.
LCMS m/z＝463.4[M/2+1]。
The seventh step: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide (Compound 8)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
8G (0.45G, 0.49mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.47G, 2.9mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 8(0.2g, 50% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.04(br，1H)，7.84-7.79(m，2H)，7.48-7.45(m，1H)，7.33-7.30(m，1H)，7.14-6.81(m，6H)，6.49-6.45(m，1H)，5.18-5.07(m，1H)，3.92-3.91(m，2H)，3.59-3.27(m，13H)，2.81-2.54(m，17H)，2.20-2.16(m，2H)，1.84-1.81(m，2H)，1.52-1.43(m，2H)，0.92-0.86(m，3H)
LCMS m/z＝811.5[M+1]。
Example 9: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] propanamide (Compound 9)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]propanamide
Figure GPA0000261095610000691
The first step is as follows: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-butyl-N- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] propanamide (9A)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]propanamide
8F (0.65g, 1.0mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.95g, 5.0mmol), stirring at 40 deg.C for 1 hr, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to obtain a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 8- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxy-ethyl radical]-5-hydroxy-4-hydro-1, 4-benzoOxazin-3-one (0.34g, 1.0mmol) was stirred at room temperature for 30 minutes, then sodium triacetoxyborohydride (0.63g, 3.0mmol) was added, and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 9A as a yellow solid (0.3g, yield 32%).
LCMS m/z＝465.4[M/2+1]。
The second step is that: 3- [2- [3- [2- [4- (1, 2-benzothiazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] propanamide (Compound 9)
3-[2-[3-[2-[4-(1，2-benzothiazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-butyl-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]propanamide
9A (0.3g, 0.32mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.29g, 1.8mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction mixture was quenched by the sequential dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 9(0.12g, 46% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ7.80-7.78(m，2H)，7.48-7.45(m，1H)，7.33-7.30(m，1H)，7.14-6.77(m，5H)，6.49-6.45(m，1H)，5.12-5.07(m，1H)，4.23-4.20(m，2H)，3.89-3.60(m，28H)，2.37-2.34(m，2H)，2.04-1.98(m，2H)，1.52-1.43(m，2H)，1.29-1.26(m，4H)，0.92-0.88(m，3H).
LCMS m/z＝816.5[M+1]。
Example 10: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 10)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000701
The first step is as follows: tert-butyl 3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionate (10A)
tert-butyl 3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
After adding 3C trifluoroacetate (0.46g, 2.0mmol) to acetonitrile (25mL), intermediate 1(0.70g, 2.0mmol), potassium carbonate (1.38g, 10.0mmol) and water (0.5mL) were added in this order, and the mixture was stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 10A as a yellow liquid (0.70g, yield 70%).
LCMS m/z＝510.3[M+1]。
The second step is that: 3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] -ethoxy ] propanoic acid (10B)
3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
10A (0.70g, 1.40mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain a yellow liquid of 10B trifluoroacetate salt (0.63g, yield 100%).
LCMS m/z＝454.2[M+1]。
The third step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanamide (10C)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 10B (0.63g, 1.40mmol) was dissolved in DCM (20mL), and triethylamine (1.43g, 7.0mmol), methylaminoacetaldehyde dimethyl acetal (0.37g, 3.1mmol), and HATU (0.79g, 2.1mmol) were added in that order and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM/methanol (v/v) ═ 15: 1) to give 10C as a yellow liquid (0.75g, yield 97.0%).
LCMS m/z＝555.5[M+1]。
The fourth step: n- [2- [ [ (2R) -2- (tert-butyl (dimethyl) silyl) oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (10D)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
10C (0.75g, 1.4mmol) was dissolved in THF (10mL), and TsOH. H was added2O (1.3g, 6.8mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.1), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.45g, 1.3mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (1.3g, 6.1mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added and extracted with DCM (100 mL. times.2), and the combined organic phases were purifiedDried over sodium sulfate, filtered, and the filtrate concentrated under reduced pressure, and the residue chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 10D as a yellow solid (0.38g, 34% yield).
LCMS m/z＝827.6[M+1]。
The fifth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 10)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
10D (0.38g, 0.46mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.15g, 0.92mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 10(0.100g, yield 31%) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.14(d，1H)，8.00(s，1H)，7.46(t，1H)，7.14(t，1H)，7.09-6.73(m，5H)，6.61(t，3H)，6.50-6.30(m，1H)，5.17(d，1H)，3.75(d，4H)，3.51(dd，8H)，3.14-2.40(m，18H)，2.00(t，3H)，1.74(t，2H).
LCMS m/z＝357.4[M/2+1]。
Example 11: 3- [2- [3- [2- [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 11)
3-[2-[3-[2-[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000721
The first step is as follows: 4- (4-Ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (11A)
tert-butyl 4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (0.512g, 2.0mmol) was added to DMF (20mL), followed by 2-chloro-3-ethylpyrimidine (0.285g, 2.0mmol), potassium carbonate (0.552g, 4.0mmol), and stirring was carried out at 50 ℃ for 4 hours. Ethyl acetate (200mL), water (100mL) and extraction were added, the aqueous phase was extracted with ethyl acetate (100mL × 1), the combined organic phases were washed with saturated brine (100mL × 1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 11A as a yellow liquid (0.667g, 92.1% yield).
LCMS m/z＝363.3[M+1]。
The second step is that: 2- (4-Ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane (11B)
4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane
11A (0.80g, 2.21mmol) was dissolved in DCM (20mL), and trifluoroacetic acid (4mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain the yellow liquid trifluoroacetate salt of 11B (0.570g, yield 98%) after concentration under reduced pressure.
The third step: tert-butyl [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionate (11C)
tert-butyl 3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 11B (0.57g, 2.17mmol) was placed in acetonitrile (25mL), and intermediate 1(0.821g, 2.30mmol), potassium carbonate (1.50g, 10.9mmol), and water (0.5mL) were added in that order. Stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 11C as a yellow liquid (1.00g, yield 86%).
LCMS m/z＝539.5[M+1]。
The fourth step: 3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (11D)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoicacid
11C (1.00g, 1.86mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 11D trifluoroacetate salt (0.90g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝483.3[M+1]。
The fifth step: n- (2, 2-dimethoxyethyl) -3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-methyl-propionamide (11E)
N-(2，2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
The trifluoroacetate salt of 11D (0.90g, 1.86mmol) was dissolved in DCM (20mL), and triethylamine (0.84g, 9.3mmol), methylaminoacetaldehyde dimethyl acetal (0.266g, 2.23mmol), and HATU (1.07g, 2.79mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM/methanol (v/v) ═ 15: 1) to give 11E as a yellow liquid (1.05g, 96% yield).
LCMS m/z＝584.5[M+1]。
And a sixth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-methyl-propionamide (11F)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
11E (1.02g, 1.750mmol) was dissolved in THF (10mL), and TsOH. H was added2O (1.66g, 8.75mmol), stirring at 40 deg.C for 1 hour, adding saturated aqueous sodium bicarbonate solution (50mL) dropwise, extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to obtain a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.586g, 1.75mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (1.12g, 5.25mmol), and stirred at room temperature for 3 hours. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (50mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 11F as a yellow solid (0.56g, 37% yield).
LCMS m/z＝428.9[M/2+1]。
The seventh step: 3- [2- [3- [2- [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionyl (Compound 11)
3-[2-[3-[2-[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
11F (0.56g, 0.65mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.21g, 1.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 11(0.16g, yield 33%).
1H NMR(400MHz，CDCl3)δ8.17(d，1H)，8.09(d，1H)，7.14(t，2H)，6.97(dd，2H)，6.89-6.79(m，1H)，6.75(s，1H)，6.61-6.48(m，1H)，6.39(d，1H)，5.14(d，1H)，3.89-3.68(m，10H)，3.66-3.52(m，5H)，3.04(s，2H)，2.94(d，3H)，2.89-2.69(m，11H)，2.65-2.55(m，5H)，1.97(t，2H)，1.81(s，2H)，1.21(dt，4H).
LCMS m/z＝742.5[M+1]。
Example 12: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 12)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000741
The first step is as follows: 4- (4-Methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (12A)
tert-butyl 4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (0.512g, 2.0mmol) was weighed into DMF (20mL), and 2-chloro-3-methylpyrimidine (0.363g, 2.0mmol) and potassium carbonate (0.552g, 4.0mmol) were added in that order, and stirred at 50 ℃ for 4 hours. Ethyl acetate (200mL), water (100mL) and the like were added to a reaction flask, followed by extraction, extraction of the aqueous phase with ethyl acetate (100mL × 1), washing of the combined organic phases with saturated brine (100mL), drying over anhydrous sodium sulfate, filtration, and concentration of the filtrate under reduced pressure, and the residue was subjected to silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to obtain 12A as a yellow liquid (0.615g, 88.4% yield).
LCMS m/z＝349.3[M+1]。
The second step is that: 4- (4-Methylpyrimidinyl-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane (12B)
4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane
12A (0.615g, 1.77mmol) was dissolved in DCM (20mL), and trifluoroacetic acid (4mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain 12B trifluoroacetate salt (0.44g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝249.3[M+1]。
The third step: 3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (12C)
tert-butyl 3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 12B (0.44g, 1.77mmol) was placed in acetonitrile (25mL), and tert-butyl 3- [2- [3- (2-bromoethyl) phenyl ] ethoxy ] propionate (intermediate 1) (0.632g, 1.77mmol), potassium carbonate (1.221g, 8.83mmol), water (0.5mL) were added in that order and stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 12C as a yellow liquid (0.82g, yield 88%).
LCMS m/z＝525.3[M+1]。
The fourth step: 3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (12D)
3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
12C (0.85g, 1.62mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 12D trifluoroacetate salt (0.76g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝469.4[M+1]。
The fifth step: n- (2, 2-dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -propionamide (12E)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
12D trifluoroacetate salt (0.76g, 1.62mmol) was dissolved in DCM (20mL), and triethylamine (0.82g, 14.1mmol), methylaminoacetaldehyde dimethyl acetal (0.232g, 1.95mmol), and HATU (0.93g, 2.43mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, and extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 12E as a yellow liquid (0.9g, yield 98%).
LCMS m/z＝570.3[M+1]。
And a sixth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-N-methyl-3- [2- [3- [ [4- (4-methylpyrimidinyl-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (12F)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
12E (0.465g, 0.82mmol) was dissolved in THF (10mL), and TsOH. H.was added2O (0.79g, 5.0mmol), stirred at 40 ℃ for 1 hour, and then saturated sodium bicarbonate water was added dropwiseThe reaction was quenched with a solution (50mL), extracted with DCM (100 mL. times.2), the combined organic phases were dried over anhydrous sodium sulfate, the crude product obtained after concentration under reduced pressure was dissolved in a mixed solvent of DCM (10mL) and methanol (3mL), and 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] added ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.267g, 0.80mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.53g, 2.46mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 12F as a yellow solid (0.56g, yield 81%).
LCMS m/z＝421.8[M/2+1]。
The seventh step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 12)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
12F (0.56g, 0.66mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.22g, 1.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 12(0.14g, yield 29%) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.13(d，1H)，8.07(s，1H)，7.13(dd，2H)，6.98(dd，2H)，6.92-6.77(m，2H)，6.46(s，1H)，6.38(d，1H)，5.17(s，1H)，3.80(d，2H)，3.72(d，6H)，3.65(s，1H)，3.57(dd，2H)，2.99-2.87(m，8H)，2.86-2.52(m，10H)，2.03(s，1H)，1.97(t，2H)，1.86(s，2H)，1.32-1.26(m，7H).
LCMS m/z＝728.5[M+1]。
Example 13: 3- [2- [4- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 13)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamid
Figure GPA0000261095610000761
The first step is as follows: [1- (cyclopropylmethyl) pyrazol-3-yl ] - [3- [2- [4- (2-hydroxyethyl) phenyl ] ethyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methanone (13A)
[1-(cyclopropylmethyl)pyrazol-3-yl]-[3-[2-[4-(2-hydroxyethyl)phenyl]ethyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methanone
The trifluoroacetate salt of 1E (0.80g, 2.0mmol) was placed in acetonitrile (25mL), and 4- (2-bromoethyl) phenethyl alcohol (0.86g, 3.8mmol), potassium carbonate (1.38g, 10.0mmol), and water (0.5mL) were added in that order. The reaction was stirred at 60 ℃ for 3.5 hours. After the reaction was cooled to room temperature, water (50mL) was sequentially added, and extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to obtain 13A as a yellow liquid (0.82g, yield 93%).
The second step is that: 3- [2- [4- [2- [8- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (13B)
tert-butyl3-[2-[4-[2-[8-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
13A (0.82g, 1.8mmol) was dissolved in acetonitrile (30mL), tert-butyl acrylate (1.0g, 7.8mmol), benzyltrimethylammonium hydroxide (0.16g, 0.96mmol, 40% solution in methanol) were added, and the mixture was stirred at room temperature for 3 hours. Most of the reaction solvent was removed by concentration under reduced pressure, and column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) gave 13B as a yellow liquid (1.01g, yield 96%).
The third step: 3- [2- [4- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (13C)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
13B (1.01g, 1.74mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 13C trifluoroacetate salt (0.88g, yield 96%) as a yellow liquid after concentration under reduced pressure.
The fourth step: 3- [2- [4- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (13D)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-methyl-propanamide
13C trifluoroacetate (0.88g, 1.7mmol) was dissolved in DCM (20mL), and triethylamine (0.85g, 8.4mmol), methylaminoacetaldehyde dimethyl acetal (0.3g, 2.5mmol), HATU (0.96g, 2.5mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM/methanol (v/v) ═ 15: 1) to give 13D as a yellow liquid (0.82g, yield 78%).
The fifth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-3- [2- [4- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (13E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
13D (0.82g, 1.3mmol) was dissolved in THF (10mL), and TsOH. H was added2O (1.2g, 6.6mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.83g, 3.9mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 13E (0.42g, yield 36%) as a yellow solid
LCMS m/z＝449.5[M/2+1]。
And a sixth step: 3- [2- [4- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 13)
3-[2-[4-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
13E (0.42g, 0.47mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (1.98g, 12.3mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 13(0.09g, 25% yield) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ8.16-8.19(dd，1H)，7.82(s，1H)，7.06-7.09(m，5H)，6.91-6.93(d，1H)，6.57(s，1H)，6.47-6.49(d，1H)，5.74(s，1H)，5.00-5.03(m，1H)，4.00-4.02(d，2H)，3.94(s，2H)，3.51-3.58(m，12H)，3.31-3.33(m，3H)，2.92(s，1H)，2.66-2.77(m，10H)，2.36-2.44(m，5H)，1.66(br，2H)1.52(br，2H)，0.51-0.52(d，2H)，0.34-0.38(m，2H)。
LCMS m/z＝784.5[M+1]。
Example 14: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 14)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000781
Figure GPA0000261095610000791
The first step is as follows: N-butyl-N- (2, 2-dimethoxyethyl) -3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (14A)
N-butyl-N-(2，2-dimethoxyethyl)-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 6B (0.42g, 0.80mmol) was dissolved in DCM (15mL), and N- (2, 2-dimethoxyethyl) butyl-1-amine (intermediate 5) (0.13g, 0.8mmol), triethylamine (0.40g, 4.0mmol), HATU (0.46g, 1.2mmol) were added in that order and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 14A as a yellow solid (0.50g, 94.0% yield).
The second step is that: N-butyl-N- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (14B)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]unde can-9-yl]ethyl]phenyl]ethoxy]propanamide
14A (0.50g, 0.75mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.71g, 3.7mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.25g, 0.75mmol) and after stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.48g, 2.2mmol) was added and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 14B as a yellow solid (0.30g, 43% yield).
LCMS m/z＝471.5[M/2+1]。
The third step: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 14)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
14B (0.30g, 0.32mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.51g, 3.2mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 14(0.06g, 20% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.05-7.91(m，2H)，7.16-6.82(m，6H)，6.51-6.43(m，1H)，5.21-5.13(m，1H)，3.74-3.29(m，20H)，2.76-2.47(m，14H)，2.20-1.67(m，8H)，0.92-0.87(m，3H)
LCMS m/z＝828.4[M+1]。
Example 15: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 15)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[8-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000801
The first step is as follows: 4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (15B)
tert-butyl 4-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (2.6g, 10.0mmol) was placed in 1, 4-dioxane (20mL), and 3-chloro-1-methyl-1H-pyrazole (15A) (1.93g, 12.0mmol), sodium tert-butoxide (2.4g, 25.0mmol), palladium acetate (0.11g, 0.5mmol), tri-tert-butylphosphine (2.0g, 1.0mmol, 10% in toluene) were added in that order and stirred at 80 ℃ for 12 hours. After the reaction was cooled to room temperature, the reaction solution was poured into saturated aqueous sodium bicarbonate (30mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2: 1) to give 15B as a yellow liquid (0.6g, yield 18%).
1H NMR(400MHz，CDCl3)δ7.15-7.14(d，1H)，5.60-5.59(m，1H)，3.83-3.81(m，4H)，3.73(s，3H)，3.20-3.11(m，4H)，2.99(s，2H)，1.98-1.95(m，2H)，1.48-1.41(m，11H)。
The second step is that: 4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane (15C)
4-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecane
15B (0.60g, 1.8mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (4mL) was added dropwise and stirred at room temperature for 4 h. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 15C trifluoroacetate salt (0.65g, yield 100%) as a yellow liquid after concentration under reduced pressure.
The third step: tert-butyl 3- [2- [3- [2- [4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionate (15D)
tert-butyl 3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 15C (0.65g, 1.8mmol) was placed in acetonitrile (15mL), and intermediate 1(0.64, 1.8mol), potassium carbonate (1.2g, 8.9mmol), water (0.5mL) were added and stirred at 60 ℃ for 24 h. Water (50mL) was added, and extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 15D as a yellow solid (0.50g, yield 55%).
LCMS m/z＝513.4[M+1]。
The fourth step: 3- [2- [3- [2- [4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (15E)
3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
15D (0.5g, 0.98mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (4mL) was added dropwise and stirred for 4 h. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 15E trifluoroacetate salt (0.45g, yield 100%) as a yellow liquid after concentration under reduced pressure.
The fifth step: N-butyl-N- (2, 2-dimethoxyethyl) -3- [2- [3- [2- [4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (15F)
N-butyl-N-(2，2-dimethoxyethyl)-3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 15E (0.45g, 1.0mmol) was dissolved in DCM (15mL), and triethylamine (0.5g, 5.0mmol), N- (2, 2-dimethoxyethyl) butyl-1-amine (intermediate 5) (0.16g, 1.0mmol), HATU (0.56g, 1.5mmol) and the mixture were added successively and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 15F as a yellow solid (0.48g, yield 81%).
LCMS m/z＝600.5[M+1]。
And a sixth step: N-butyl-N- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -propionamide (15G)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
15F (0.48g, 0.8mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.76g, 4.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.27g, 0.8mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.51g, 2.4mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 15G as a yellow solid (0.28G, 40% yield).
LCMS m/z＝436.9[M/2+1]。
The seventh step: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (1-methylpyrazol-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 15)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[8-(1-methylpyrazol-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
15G (0.28G, 0.32mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.31G, 1.9mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 15(0.1g, 40% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.09-8.07(m，1H)，6.96-6.76(m，8H)，6.54-6.45(m，1H)，4.30-3.75(m，15H)，3.30-2.59(m，21H)，2.11-2.08(m，2H)，1.90-1.86(m，2H)，143-1.24(m，2H)，0.92-0.86(m，3H)。
LCMS m/z＝758.5[M+1]。
Example 16: 3- [2- [3- [2- [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 16)
3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000821
The first step is as follows: 2-chloro-4-ethyl-pyridine (16B)
2-chloro-4-ethyl-pyridine
2-amino-4-ethylpyridine (16A) (8.0g, 65mmol) was dissolved in 100mL of concentrated hydrochloric acid, and an aqueous solution (40mL) of sodium nitrite (6.8g, 98mmol) and an aqueous solution (40mL) of sodium chloride (7.7g, 130mmol) were added dropwise in this order at 0 ℃ and, after completion of the addition, stirring was continued for 30 minutes. The reaction solution was made alkaline with a 20% aqueous solution of sodium hydroxide, extracted with ethyl acetate (150mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 15: 1) to give 16B as a yellow liquid (6.5g, yield 70%).
1H NMR(400MHz，CDCl3)δ8.26-8.24(d，1H)，7.16(s，1H)，7.05-7.03(m，1H)，2.67-2.61(m，2H)，1.26-1.22(m，3H).
The second step is that: 4- (4-Ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (16C)
tert-butyl 4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (5.12g, 20.0mmol) was dissolved in 1, 4-dioxane (50mL), and 16B (4.24g, 30.0mmol), cesium carbonate (13.0g, 40.0mmol), 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene (1.6g, 2.8mmol), palladium acetate (0.3g, 1.4mmol) and a nitrogen atmosphere were added in this order to conduct a reflux reaction for 8 hours. Water (100mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 16C as a yellow liquid (3.0g, yield 42%).
LCMS m/z＝362.3[M+1]。
The third step: 4- (4-Ethyl-2-pyridine) -1-oxa-4, 9-diazaspiro [5.5] undecane (16D)
4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane
16C (0.5g, 1.4mmol) was dissolved in DCM (30mL), and trifluoroacetic acid (6mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain 16D trifluoroacetate salt (0.52g, yield 100%) as a yellow liquid after concentration under reduced pressure.
The fourth step: tert-butyl 3- [2- [3- [2- [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undec-9-yl ] ethyl ] phenyl ] ethoxy ] propionate (16E)
tert-butyl 3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 16D (0.52g, 1.4mmol) was placed in acetonitrile (15mL), and intermediate 1(0.49g, 1.4mmol), potassium carbonate (0.95g, 6.9mmol), and water (0.5mL) were added in that order and stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 16E as a yellow liquid (0.6g, yield 80%).
The fifth step: 3- [2- [3- [2- [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undec-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (16F)
3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
16E (0.78g, 1.5mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 16F trifluoroacetate salt (0.7g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝482.4[M+1]。
And a sixth step: n- (2, 2-Dimethoxyethyl) -3- [2- [3- [2- [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-methyl-propionamide (16G)
N-(2，2-dimethoxyethyl)-3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]unde can-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
The trifluoroacetic acid salt of 16F (0.7g, 1.5mmol) was dissolved in DCM (20mL), and triethylamine (0.74g, 7.3mmol), methylaminoacetaldehyde dimethyl acetal (0.26g, 2.2mmol), HATU (0.83g, 2.2mmol) and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 16G as a yellow solid (0.55G, yield 65%).
The seventh step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-methyl-propionamide (16H)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-methyl-propanamide
16G (0.6G, 1.0mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.96g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.64g, 3.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 16H as a yellow solid (0.55g, yield 60%).
LCMS m/z＝428.4[M/2+1]。
The seventh step: 3- [2- [3- [2- [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 16)
3-[2-[3-[2-[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
16H (0.55g, 0.64mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (1.0g, 6.4mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 16(0.3g, yield 60%) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.10-8.05(m，2H)，7.20-7.16(m，2H)，6.97-6.94(m，2H)，6.83-6.80(m，1H)，6.70(s，1H)，6.60-6.52(m，2H)，6.47(s，1H)，5.14-5.09(m，1H)，3.81-3.37(m，13H)，3.03-2.54(m，21H)，2.05-1.98(m，2H)，1.78(br，2H)，1.28-1.20(m，3H).
LCMS m/z＝371.4[M/2+1]。
Example 17: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [4- [ [4- (2-pyridinyl-1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 17)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000851
The first step is as follows: 3- [2- [4- [ [4- (2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (17B)
tert-butyl 3-[2-[4-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
17A (0.35g, 1.50mmol) and tert-butyl 3- [2- (4-formylphenyl) ethoxy ] propionate (4d) (0.42g, 1.51mmol) were dissolved in methanol (16mL) solvent, stirred at room temperature for 1 hour, sodium triacetoxyborohydride (0.95g, 4.5mmol) was added, stirred at room temperature for 3 hours, a saturated sodium bicarbonate solution (10mL) was added to quench the reaction, DCM (20mL) was added, the layers were extracted, the aqueous phase was extracted with DCM (50 mL. times.2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was isolated and purified by column chromatography on silica gel to give 17B as a yellow liquid (0.342g, 46% yield).
LCMS m/z＝496.3[M+1]。
The second step is that: 3- [2- [4- [ [4- (2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (17C)
3-[2-[4-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
Dissolve 17B (0.342g, 0.69mmol) in DCM (15mL) and stir with trifluoroacetic acid (5mL) dropwise at RT for 4 h. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 17C trifluoroacetate salt (0.309g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝440.4[M+1]。
The third step: n- (2, 2-dimethoxyethyl) -N-methyl-3- [2- [4- [ [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (17D)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 17C (0.303g, 0.69mmol) was dissolved in DCM (20mL), and triethylamine (0.25g, 3..45mmol), methylaminoacetaldehyde dimethyl acetal (0.098g, 0.82mmol), and HATU (0.393g, 1.04mmol) were added in that order and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 17D as a yellow liquid (0.37g, yield 99.3%).
LCMS m/z＝541.3[M+1]。
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-N-methyl-3- [2- [4- [ [4- (2-pyridinyl) -1-oxo-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (17E)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
17D (0.372g, 0.688mmol) was dissolved in THF (10mL) and TsOH. H was added2O (0.66g, 3.44mmol), stirring at 40 ℃ for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a residue, dissolving the residue in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.23g, 0.688mmol) and, after stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.43g, 2.0mmol) was added and, after stirring at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 17E as a yellow solid (0.30g, yield 553.6%).
LCMS m/z＝449.8[M/2+1]。
The fifth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [4- [ [4- (2-pyridinyl-1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 17)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[4-[[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
17E (0.30g, 0.369mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.12g, 0.738mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 17(0.080g, 31% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.12(s，2H)，7.49(d，1H)，7.30(s，1H)，7.20-6.93(m，3H)，6.82(s，1H)，6.60(s，3H)，5.48(d，2H)，3.76(s，7H)，3.53(m，10H)，3.35(s，3H)，3.07(s，2H)，3.03(d，1H)，2.94(s，3H)，2.80(s，2H)，2.63(s，3H)，2.01(d，4H)，1.87(s，2H).
LCMS m/z＝700.5[M+1]。
Example 18: 3- [2- [3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-azaspirodiaza [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 18)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000861
Figure GPA0000261095610000871
The first step is as follows: 3- [2- [3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (18A)
tert-butyl 3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The 16D trifluoroacetate salt (0.52g, 1.5mmol) was placed in acetonitrile (15mL), and intermediate 2(0.53g, 1.5mmol), potassium carbonate (1.1g, 7.7mmol) and water (0.5mL) were added in that order and stirred at room temperature for 8 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then the residue was separated by silica gel column chromatography (DCM/methanol (v/v) ═ 15: 1) to obtain 18A as a yellow liquid (0.5g, yield 65%).
LCMS m/z＝524.5[M+1].
The second step is that: 3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (18B)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
18A (0.5g, 1.0mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain the trifluoroacetate salt of 18B (0.45g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝468.4[M+1].
The third step: n- (2, 2-Dimethoxyethyl) -3- [2- [3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (18C)
N-(2，2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
The trifluoroacetate salt of 18B (0.45g, 1.0mmol) was dissolved in DCM (20mL), and triethylamine (0.50g, 5.0mmol), methylaminoacetaldehyde dimethyl acetal (0.17g, 1.4mmol), and HATU (0.55g, 1.4mmol) were added successively and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM/methanol (v/v) ═ 15: 1) to give 18C as a yellow solid (0.50g, 91% yield).
LCMS m/z＝570.5[M+1]。
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (18D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
18C (0.5g, 0.88mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.84g, 4.4mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.29g, 0.88mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.56g, 2.6mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographed on silica gel (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 18D as a yellow solid (0.55g, 74% yield).
LCMS m/z＝422.15[M/2+1]。
The fifth step: 3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 18)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
18D (0.55g, 0.65mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (1.0g, 6.4mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the successive dropwise addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was chromatographed on a silica gel column (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 18(0.3g, 60% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.44-8.41(d，1H)，7.91-7.90(d，1H)，7.39-7.33(m，5H)，7.20(s，1H)，7.06-7.04(d，1H)，6.97-6.95(d，1H)，6.69-6.66(d，1H)，5.43-5.40(m，1H)，4.32(s，2H)，3.97-3.95(m，2H)，3.75-3.62(m，11H)，3.33-3.20(m，7H)，3.11(s，3H)，2.90-2.88(m，2H)，2.78-2.67(m，4H)，2.27-2.23(m，2H)，1.93-1.89(m，2H)，1.32-1.28(m，3H).
LCMS m/z＝728.5[M+1]。
Example 19: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 19)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000881
Figure GPA0000261095610000891
The first step is as follows: tert-butyl 3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoate (19B)
tert-butyl 3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 12B (0.50g, 2.01mmol) was placed in acetonitrile (25mL), and tert-butyl 3- [2- [3- (chloromethyl) phenyl ] ethoxy ] propionate (intermediate 6) (0.602g, 2.01mmol), potassium carbonate (1.39g, 10.1mmol), water (0.5mL) and stirred at 60 ℃ for 24 hours were added sequentially. After the reaction was cooled to room temperature, water (50mL) was added, and extraction was performed with ethyl acetate (100mL × 2), and the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 19B as a yellow liquid (0.82g, yield 88%).
LCMS m/z＝511.3[M+1]。
The second step is that: 3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (19C)
3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
Dissolve 19B (1.0g, 1.98mmol) in DCM (15mL) and stir with trifluoroacetic acid (5mL) dropwise at room temperature for 4 h. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 19C trifluoroacetate salt (0.89g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝455.3[M+1].
The third step: n- (2, 2-dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -propionamide (19D)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
The 19C trifluoroacetate salt (0.45g, 0.98mmol) was dissolved in DCM (20mL), and triethylamine (0.99g, 9.8mmol), methylaminoacetaldehyde dimethyl acetal (0.14g, 1.2mmol), and HATU (0.56g, 1.5mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 19D as a yellow liquid (0.54g, 98% yield).
LCMS m/z＝556.3[M+1]。
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-N-methyl-3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (19E)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
19D (0.540g, 0.97mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.95g, 5.0mmol), stirring at 40 ℃ for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] ester]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.325g, 0.97mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.62g, 2.91mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 19E as a yellow solid (0.40g, 50% yield).
LCMS m/z＝829.5[M+1]。
The fifth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 19)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
19E (0.40g, 0.48mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.14g, 0.72mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 19(0.11g, 59% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.40(d，1H)，8.19(d，1H)，7.42-7.24(m，5H)，7.03(d，1H)，6.75-6.54(m，2H)，5.40(d，1H)，4.86(dd，14H)，3.87-3.64(m，12H)，2.89(t，2H)，2.67(t，2H)，2.39(s，3H)，2.19(d，2H)，1.79(s，2H).
LCMS m/z＝715.5[M+1]。
Example 20: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-ethyl-3- [2- [3- [2- [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 20)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-ethyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000901
Figure GPA0000261095610000911
The first step is as follows: n- (2, 2-dimethoxyethyl) -N-ethyl-3- [2- [3- [ [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -propionamide (20B)
N-(2，2-dimethoxyethyl)-N-ethyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
12D trifluoroacetate (0.525g, 1.12mmol) was dissolved in DCM (20mL), and triethylamine (0.34g, 3.36mmol), ethylaminoacetaldehyde dimethyl acetal (0.179g, 1.34mmol), and HATU (0.852g, 2.24mmol) were added in that order and stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 20B as a yellow liquid (0.65g, 98% yield).
LCMS m/z＝584.4[M+1].
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-N-ethyl-3- [2- [3- [ [4- (4-methylpyrimidinyl-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (20C)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-ethyl-3-[2-[3-[[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
20B (0.65g, 1.12mmol) was dissolved in THF (10mL), and TsOH. H was added2O (1.09g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.375g, 1.12mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.712g, 3.36mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 20C as a yellow solid (0.384g, 40% yield).
LCMS m/z＝856.5[M+1]。
The third step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-ethyl-3- [2- [3- [2- [4- (4-methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 20)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-ethyl-3-[2-[3-[2-[4-(4-methylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
20C (0.38g, 0.45mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.18g, 0.9mmol) was added, and the mixture was stirred at room temperature for 12 hours. To this, saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 20(0.12g, yield 36%) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.40(d，1H)，8.17(d，1H)，7.40-7.29(m，5H)，7.03(d，1H)，6.63-6.65(m，2H)，5.44(d，1H)，4.29(dd，2H)，3.68-3.81(m，13H)，3.20-3.45(m，12H)，2.89(t，2H)，2.65(t，2H)，2.38(s，3H)，2.15-2.18(m，2H)，1.85(s，2H).1.18-1.41(m，6H).
LCMS m/z＝742.5[M+1]。
Example 21: 3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 21)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000921
The first step is as follows: tert-butyl 3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoate (21B)
tert-butyl 3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 11B (0.538g, 2.05mmol) was placed in acetonitrile (25mL), and tert-butyl 3- [2- [3- (2-chloromethyl) phenyl ] ethoxy ] propionate (intermediate 6) (0.613g, 2.05mmol), potassium carbonate (1.42g, 10.3mmol), water (0.5mL) and stirring at 60 ℃ for 24 hours were added sequentially. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous ammonium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 21B as a yellow liquid (0.791g, 73.5% yield).
LCMS m/z＝525.3[M+1]。
The second step is that: 3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (21C)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
21B (0.37g, 0.71mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 21C trifluoroacetate salt (0.33g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝469.3[M+1].
The third step: n- (2, 2-dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -propionamide (21D)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
The trifluoroacetic acid salt of 21C (0.37g, 0.79mmol) was dissolved in DCM (20mL), and triethylamine (0.79g, 7.8mmol), methylaminoacetaldehyde dimethyl acetal (0.11g, 0.95mmol), and HATU (0.45g, 1.2mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 21D as a yellow liquid (0.44g, 98% yield).
LCMS m/z＝571.3[M+1]。
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-N-methyl-3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (21E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
21D (0.370g, 0.65mmol) was dissolved in THF (10mL), and TsOH. H.was added thereto2O (0.62g, 3.2mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.22g, 0.65mmol) and, after stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.41g, 1.91mmol) was added and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 21E as a yellow solid (0.22g, 40% yield).
LCMS m/z＝842.4[M+1]。
The fifth step: 3- [2- [3- [ [4- (4-ethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 21)
3-[2-[3-[[4-(4-ethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
21E (0.215g, 0.255mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.074g, 0.38mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 21(0.11g, 59% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.42(d，1H)，8.16(d，1H)，7.48-7.19(m，5H)，7.03(d，1H)，6.66(d，1H)，6.50(d，1H)，5.42-5.30(m，1H)，4.11(s，2H)，3.88-3.58(m，11H)，3.47(s，1H)，3.27(s，1H)，3.20(dq，4H)，3.16-2.99(m，8H)，2.90(dd，2H)，2.72-2.63(m，3H)，2.59(dd，2H)，2.04(dd，3H)，1.87-1.68(m，3H)，1.58(s，2H).
LCMS m/z＝729.4[M+1]。
Example 22: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-hetero spiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (compound 22)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000941
The first step is as follows: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (22A)
tert-butyl 3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 1E (0.95g, 3.1mmol) was placed in acetonitrile (25mL), and tert-butyl 3- [2- [3- (chloromethyl) phenyl ] ethoxy ] propionate (intermediate 6) (0.93g, 3.1mmol), potassium carbonate (1.3g, 9.4mmol), water (1mL) were added in that order and stirred at 60 ℃ for 3.5 hours. After the reaction was cooled to room temperature, water (50mL) was added, and extraction was performed with DCM (100mL × 2), and the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 22A as a yellow liquid (1.65g, 93% yield).
The second step is that: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (22B)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-methyl-propanamide
22A (1.65g, 2.9mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (5mL) was added dropwise at 0 ℃ to complete the reaction at room temperature for 1 hour. The reaction mixture was directly concentrated under reduced pressure, and toluene (20mL) was added to the reaction mixture, followed by concentration under reduced pressure to obtain a residue. DCM (15mL) and triethylamine (1.37g, 13.5mmol) were added to the residue in this order at 0 ℃ under nitrogen, and after stirring well, N- (2, 2-dimethoxyethyl) methyl-1-ammonia (0.39g, 3.2mmol) and HATU (1.54g, 4.0mmol) were added and reacted at room temperature for 1 hour. Water (30mL) was added, DCM (30mL × 2) was extracted, the organic phases were combined, washed with brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 22B as a yellow liquid (1.51g, 91% yield).
The third step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (5-hydroxy-3-oxa-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (22C)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
22B (0.75g, 1.2mmol) was dissolved in THF (15mL), and TsOH. H.was added2O (1.2g, 6.1mmol), stirred at 40 ℃ for 1 hour, quenched by dropwise addition of saturated aqueous sodium bicarbonate (50mL), and quenched with DCM (100 mL. times.2), the combined organic phases were dried over anhydrous sodium sulfate, and the crude product obtained after concentration under reduced pressure was dissolved in a mixed solvent of DCM (10mL) and methanol (3mL), and 8- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] added]Oxy-ethyl radical]-5-hydroxy-4H-1, 4-benzoxazin-3-one (0.26g, 0.86mmol), stirring at room temperature for 30 minutes, adding sodium triacetoxyborohydride (0.78g, 3.7mmol), and stirring at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 22C as a yellow solid (0.52g, 48% yield).
The fourth step: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (compound 22)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide
22C (0.52g, 0.59mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.98g, 6.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 22(0.22g, 49% yield) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ9.80(br，1H)，7.82(d，1H)，7.19(m，1H)，7.08-7.10(m，3H)，6.86(dd，1H)，6.55(s，1H)，6.49(d，1H)，4.99(m，1H)，4.84-4.78(m，1H)，4.47(t，2H)，3.91-4.01(m，4H)，3.67-3.51(m，8H)，3.22-3.42(m，8H)，2.75-2.78(m，4H)，2.67(t，1H)，2.61(dd，2H)，2.37-2.24(m，4H)，1.68(d，2H)，1.45-1.57(m，2H)，0.49-0.57(m，2H)，0.32-0.41(m，2H).
LCMS m/z＝774.4[M+1].
Example 23: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (Compound 23)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610000951
Figure GPA0000261095610000961
The first step is as follows: 4- (4-methyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (23B)
tert-butyl 4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (5.12g, 20.0mmol) was placed in 1, 4-dioxane (50mL), and 2-chloro-4-methyl-pyridine (23A) (3.83g, 30.0mmol), cesium carbonate (13.0g, 40.0mmol), 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene (1.6g, 2.8mmol), palladium acetate (0.3g, 1.4mmol), refluxed under nitrogen for 8 hours. The reaction mixture was cooled to room temperature, water (100mL) was added thereto, and extraction was performed with ethyl acetate (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 23B as a yellow liquid (3.0g, yield 43%).
The second step is that: 3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (23C)
tert-butyl 3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
After 23B (1.02g, 2.9mmol) was added to DCM (20mL) and trifluoroacetic acid (3mL) was added dropwise at 0 ℃ to complete the reaction at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure, and toluene (20mL) was added and concentrated again under reduced pressure. The concentrated residue was taken up in acetonitrile (25mL), and intermediate 6(0.98g, 3.3mmol), potassium carbonate (1.5g, 11.0mmol) and water (1mL) were added in that order and stirred at 60 ℃ for 3.5 hours. After the reaction was cooled to room temperature, water (50mL) was added, and extraction was performed with DCM (100mL × 2), and the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 23C as a yellow liquid (1.42g, yield 97%).
The third step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (23D)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
23C (1.42g, 2.79mmol) was added to DCM (20mL), trifluoroacetic acid (4mL) was added dropwise at 0 ℃ and after completion of the dropwise addition, the reaction was allowed to react at room temperature for 1 hour, and then the system was concentrated under reduced pressure, and toluene (20mL) was added and concentrated again under reduced pressure. DCM (15mL) and triethylamine (1.39g, 13.8mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) methyl-1-ammonia (0.43g, 3.6mmol) and HATU (1.57g, 4.1mmol) were added and the mixture was reacted at room temperature for 1 hour. After completion of the reaction, water (30mL) was added, and extraction was performed with DCM (80mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 23D as a yellow liquid (1.11g, yield 73%).
LCMS m/z＝555.3[M+1]。
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (23E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
23D (0.55g, 1.0mmol) was dissolved in THF (15mL), and TsOH. H.was added thereto2O (0.94g, 5.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.23g, 0.69mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.63g, 3.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 23E as a yellow solid (0.23g, 28% yield).
The fifth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (Compound 23)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
23E (0.23g, 0.28mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.8g, 5.0mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 23(0.14g, yield 70.8%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.29(s，1H)，8.17(d，1H)，7.94(d，1H)，7.23-7.16(m，1H)，7.14(s，1H)，7.05-7.11(m，3H)，6.91(d，1H)，6.65(s，1H)，6.51-6.44(m，2H)，5.01(t，1H)，3.69-3.65(m，2H)，3.62-3.54(m，4H)，3.44-3.23(m，12H)，2.75-2.77(m，3H)，2.73-2.60(m，4H)，2.41-2.44(m，2H)，2.28(t，2H)，2.20(s，3H)，1.78-1.70(m，2H)，1.54(dt，2H).
LCMS m/z＝357.3[M/2+1].
Example 24: 3- [2- [4- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 24)
3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610000971
Figure GPA0000261095610000981
The first step is as follows: 3- [2- [4- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (24A)
tert-butyl 3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 1E (0.38g, 1.2mmol) was placed in acetonitrile (25mL), and tert-butyl 3- [2- [4- (bromomethyl) phenyl ] ethoxy ] propionate (intermediate 2) (0.45g, 1.3mmol), potassium carbonate (0.35g, 2.5mmol), water (1mL) were added in that order and stirred at 60 ℃ for 3.5 hours. After the reaction was cooled to room temperature, water (50mL) was added, and extraction was performed with DCM (100mL × 2), and the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 24A as a yellow liquid (0.48g, yield 68%).
The second step is that: 3- [2- [4- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (24B)
3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-methyl-propanamide
After 24A (0.48g, 0.85mmol) was added to DCM (10mL) and trifluoroacetic acid (2mL) was added dropwise at 0 ℃ and reacted at room temperature for 1 hour, the system was concentrated under reduced pressure, and toluene (20mL) was added and concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.41g, 4.0mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) methyl-1-ammonia (0.14g, 1.2mmol) and HATU (0.46g, 1.2mmol) were added and the mixture was allowed to warm to room temperature for 1 hour. Water (30mL) was added to the reaction solution, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 24B as a yellow liquid (0.42g, yield 85%).
The third step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [4- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (24C)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
24B (0.42g, 0.69mmol) was dissolved in THF (15mL), and TsOH. H.was added thereto2O (0.98g, 3.6mmol) was stirred at 40 deg.CAfter stirring for 1 hour, the reaction was quenched by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extracted with DCM (100 mL. times.2), the combined organic phases were dried over anhydrous sodium sulfate, and the crude product obtained after concentration under reduced pressure was dissolved in a mixed solvent of DCM (10mL) and methanol (3mL), and 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] added]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.18g, 0.55mmol) and, after stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.44g, 2.1mmol) was added and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 24C as a yellow solid (0.23g, 37% yield).
LCMS m/z＝442.9[M/2+1]。
The fourth step: 3- [2- [4- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 24)
3-[2-[4-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
24C (0.23g, 0.30mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.99g, 6.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. To this, saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 24(0.11g, yield 49%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.22(br，1H)，8.17(d，1H)，7.82(d，1H)，7.14(m，4H)，7.06(d，1H)，6.91(d，1H)，6.55(br，1H)，6.48(dd，1H)，5.00(dd，1H)，4.01(m，2H)，3.90-3.94(m，2H)，3.66-3.52(m，8H)，3.42-3.23(m，9H)，2.78-2.71(m，4H)，2.62-2.69(m，3H)，2.26-2.33(m，4H)，1.64-1.68(m，2H)，1.48(m，2H)，0.50-0.57(m，2H)，0.40-0.34(m，2H).
LCMS m/z＝770.3[M+1]。
Example 25: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-ethyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide (Compound 25)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
Figure GPA0000261095610000991
The first step is as follows: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-ethyl-propionamide (25A)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-ethyl-propanamide
After 24A (0.39g, 0.69mmol) was added to DCM (10mL) and trifluoroacetic acid (5mL) was added dropwise at 0 ℃ and reacted at room temperature for 1 hour, the system was concentrated under reduced pressure, and toluene (20mL) was added and concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.45g, 4.4mmol) were added to the residue in that order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) ethane-1-ammonia (0.35g, 2.6mmol) and HATU (0.60g, 1.6mmol) were added and the mixture was allowed to warm to room temperature for 1 hour. After completion of the reaction, water (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to obtain 25A (0.37g, yield 88%) as a yellow liquid.
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-ethyl-propionamide (25B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-ethyl-propanamide
25A (0.37g, 0.58mmol) was dissolved in THF (15mL), and TsOH. H.was added2O (0.56g, 2.9mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.16g, 0.47mmol) and after stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.37g, 1.75mmol) was added and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 25B as a yellow solid (0.27g, 51% yield).
LCMS m/z＝449.9[M/2+1].
The third step: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-ethyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide (Compound 25)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
25B (0.27g, 0.30mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.80g, 5.0mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 25(0.10g, yield 44%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.22(br，1H)，8.17(d，1H)，7.82(d，1H)，7.04-7.18(m，5H)，6.91(d，1H)，6.55(s，1H)，6.50-6.45(m，1H)，4.99-5.00(m，1H)，4.01(s，2H)，3.91-3.93(m，2H)，3.66-3.50(m，8H)，3.43-3.18(m，11H)，2.87-2.56(m，7H)，2.27-2.33(m，4H)，1.67(d，2H)，1.44-1.55(m，2H)，0.49-0.57(m，2H)，0.33-0.40(m，2H).
LCMS m/z＝784.3[M+1].
Example 26: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-propyl-propionamide (Compound 26)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-propyl-propanamide
Figure GPA0000261095610001011
The first step is as follows: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-propyl-propionamide (26A)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-propyl-propanamide
24A (0.39g, 0.63mmol) was added to DCM (10mL), trifluoroacetic acid (2mL) was added dropwise at 0 ℃ and after completion of the addition, the reaction was carried out at room temperature for 1 hour, then the system was concentrated under reduced pressure, toluene (20mL) was added and the reaction was concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.45g, 4.4mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) propan-1-amine (0.36g, 2.4mmol) and HATU (0.60g, 1.6mmol) were added and the mixture was allowed to warm to room temperature for 1 hour. After completion of the reaction, water (30mL) was added, extraction was performed with DCM (50mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 26A as a yellow liquid (0.36g, yield 84%).
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-propyl-propionamide (26B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-propyl-propanamide
26A (0.36g, 0.56mmol) was dissolved in THF (15mL), and TsOH. H.was added2O (0.53g, 2.8mmol), stirring at 40 ℃ for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, which is dissolved in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.15g, 0.45mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.36g, 1.7mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 26B as a yellow solid (0.20g, 39% yield).
LCMS m/z＝456.7[M/2+1]。
The third step: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-propyl-propionamide (Compound 26)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-propyl-propanamide
26B (0.20g, 0.22mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.98g, 6.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 26(0.13g, yield 72%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.24(s，1H)，8.18(d，1H)，7.82(d，1H)，7.04-7.09(m，5H)，6.91(d，1H)，6.55(s，1H)，6.50-6.45(m，1H)，5.04-4.94(m，1H)，4.01(m，2H)，3.91-3.94(m，2H)，3.50-3.61(m，8H)，3.46-3.22(m，14H)，2.78-2.59(m，6H)，2.28-2.33(m，4H)，1.67(d，2H)，1.45-1.49(m，2H)，0.50-0.58(m，2H)，0.33-0.41(m，2H).
LCMS m/z＝798.5[M+1].
Example 27: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-ethyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide (Compound 27)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
Figure GPA0000261095610001021
The first step is as follows: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-ethyl-propionamide (27A)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-ethyl-propanamide
1F (0.71g, 1.2mmol) was added to DCM (10mL), trifluoroacetic acid (2mL) was added dropwise at 0 ℃ and after completion of the addition, the reaction was carried out at room temperature for 1 hour, then the system was concentrated under reduced pressure, toluene (20mL) was added and the reaction was concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.59g, 5.8mmol) were added to the residue in that order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) ethane-1-ammonia (0.17g, 1.3mmol) and HATU (0.66g, 1.7mmol) were added and the mixture was allowed to warm to room temperature for 1 hour. After completion of the reaction, water (30mL) was added, and extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to obtain 27A as a yellow liquid (0.70g, yield: 94%).
LCMS m/z＝640.4[M+1]。
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-ethyl-propionamide (27B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-ethyl-propanamide
27A (0.37g, 0.58mmol) was dissolved in THF (15mL), to which TsOH. H.was added2O (0.61g, 3.2mmol), stirred at 40 ℃ for 1 hour, quenched by dropwise addition of saturated aqueous sodium bicarbonate (50mL), and quenched with DCM (100 mL. times.2)) The extract was extracted, the combined organic phases were dried over anhydrous sodium sulfate, and the crude product obtained after concentration under reduced pressure was dissolved in a mixed solvent of DCM (10mL) and methanol (3mL), and 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group was added]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.2g, 0.6mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.37g, 1.7mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 27B as a yellow solid (0.20g, 38% yield).
LCMS m/z＝456.9[M/2+1]。
The third step: 3- [2- [3- [2- [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] -N-ethyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide (Compound 27)
3-[2-[3-[2-[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]-N-ethyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide
27B (0.20g, 0.22mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.99g, 6.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. To this, saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 27(0.125g, yield 71%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.27(br，1H)，8.18(dd，1H)，7.82(s，1H)，7.17-7.11(m，1H)，6.99-7.08(m，4H)，6.92(dd，1H)，6.57(s，1H)，6.49(dd，1H)，4.99-5.09(m，1H)，4.01(d，2H)，3.94(s，2H)，3.67-3.50(m，9H)，3.18-3.42(m，9H)，2.78-2.59(m，11H)，2.31-2.43(m，4H)，1.63-1.71(m，2H)，1.48-1.57(m，2H)，0.49-0.55(m，2H)，0.38-0.34(m，2H).
LCMS m/z＝798.5[M+1]。
Example 28: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethylamino ] ethyl ] -N-methyl-propionamide bistrifluoroacetate (Compound 28)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethylamino]ethyl]-N-methyl-propanamide ditrifluoroaceticacid
Figure GPA0000261095610001041
24B (0.37g, 0.60mmol) was dissolved in N-methylpyrrolidinone (15mL) and the acetate salt of 8- (2-aminoethyl) -5-hydroxy-4H-1, 4-benzoxazin-3-one (0.15g, 0.72mmol), acetic acid (0.037g, 0.61mmol) were added sequentially. After stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.37g, 1.7mmol) was added and the reaction was terminated after 5 hours. Most of the reaction solvent was removed by concentration under reduced pressure, a saturated aqueous sodium bicarbonate solution (50mL) was slowly dropped into the residue, extraction was performed with DCM (100mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, and then subjected to column chromatography separation under reduced pressure to give a crude product [ (DCM/methanol (v/v) ═ 15: 1) ], which was separated with a liquid-phase preparative column (liquid-phase preparative condition: C18 reverse-phase preparative column, mobile phases were deionized water (a) containing 0.1% trifluoroacetic acid, acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution B (a + B) ═ 5% to 20%, elution time 15min, flow rate 12mL/min, column temperature: 30 ℃) to give compound 28(0.034g, yield 7%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ9.78(br，1H)，7.82(d，1H)，7.22-7.16(m，1H)，7.14-7.05(m，3H)，6.60(d，1H)，6.55(s，1H)，6.45-6.38(m，1H)，4.46(d，2H)，4.04-3.98(m，2H)，3.96-3.87(m，2H)，3.65-3.50(m，9H)，3.43-3.26(m，11H)，2.66-2.53(m，6H)，2.38-2.25(m，4H)，1.70-1.63(m，2H)，1.56-1.46(m，2H)，0.57-0.50(m，2H)，0.40-0.34(m，2H).
LCMS m/z＝758.4[M+1]。
Example 29: n-cyclopentyl-3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethylamino ] ethyl ] propionamide ditrifluoroacetate (Compound 29)
N-cyclopentyl-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethylamino]ethyl]propanamideditrifluoroacetic acid
Figure GPA0000261095610001042
The first step is as follows: n-cyclopentyl-3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) propanamide (29A)
N-cyclopentyl-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)propanamide
24A (0.39g, 0.67mmol) was added to DCM (10mL), trifluoroacetic acid (2mL) was added dropwise at 0 ℃ and after completion of the addition, the reaction was carried out at room temperature for 1 hour, then the system was concentrated under reduced pressure, toluene (20mL) was added and the reaction was concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.45g, 4.4mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) cyclopentylamine (0.40g, 2.3mmol) and HATU (0.60g, 1.6mmol) were added and the mixture was allowed to warm to room temperature for 1 hour. After completion of the reaction, water (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 29A as a yellow liquid (0.40g, yield 91%).
The second step is that: n-cyclopentyl-3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethylamino ] ethyl ] propionamide ditrifluoroacetate (Compound 29)
N-cyclopentyl-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethylamino]ethyl]propanamide ditrifluoroacetic acid
29A (0.40g, 0.61mmol) was dissolved in N-methylpyrrolidone (15mL) and the acetate salt of 8- (2-aminoethyl) -5-hydroxy-4H-1, 4-benzoxazin-3-one (0.15g, 0.72mmol), acetic acid (0.04g, 0.7mmol) were added in that order. After stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.40g, 1.9mmol) was added and the reaction was terminated after 5 hours. Most of the reaction solvent was removed by concentration under reduced pressure, a saturated aqueous sodium bicarbonate solution (50mL) was slowly dropped into the residue, extraction was performed with DCM (100mL × 2), the combined organic phases were washed with a saturated saline solution (50mL), dried over anhydrous sodium sulfate, and the residue after concentration under reduced pressure was purified by silica gel column chromatography (DCM/methanol (v /): 1: 10) to obtain a crude product, which was prepared by column chromatography (liquid phase preparation condition: C18 reverse phase preparation column, mobile phase was deionized water (a) containing 0.1% trifluoroacetic acid, mobile phase was acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution B (a + B): 5% to 20%, elution time 15min, flow rate 12mL/min, column temperature: 30 ℃) to obtain compound 29 as a white solid (0.029g, yield 6%).
1H NMR(400MHz，CD3OD)δ7.71(d，1H)，7.40-7.31(m，4H)，6.73(d，1H)，6.63(d，1H)，6.49(d，1H)，4.62(s，2H)，4.30-4.26(m，2H)，4.09-3.98(m，4H)，3.82-3.73(m，3H)，3.70(dd，4H)，3.65-3.62(m，1H)，3.50(t，2H)，3.39-3.31(m，2H)，3.25-3.16(m，4H)，3.12(t，2H)，2.93-2.87(m，4H)，2.68(t，2H)，2.24-2.16(m，2H)，1.91-1.83(m，2H)，1.81-1.70(m，4H)，1.64-1.57(m，2H)，1.53-1.45(m，2H)，0.99-0.82(m，2H)，0.59(d，2H)，0.38(s，2H).
LCMS m/z＝812.5[M+1]。
Example 30: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-methoxypyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (Compound 30)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
Figure GPA0000261095610001061
The first step is as follows: 4- (4-Methylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (30B)
tert-butyl 4-(4-methoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (0.769g, 3.00mmol) was dissolved in DMF (20mL), and 2-bromo 3-methoxypyrimidine (30A) (0.567g, 3.00mmol) and potassium carbonate (0.829g, 6.00mmol) were added in this order and stirred at 90 ℃ for 4 hours. Ethyl acetate (200mL), water (100mL) was added thereto, extraction was performed, the organic phase and the aqueous phase were separated, the aqueous phase was extracted with ethyl acetate (100mL × 1), the combined organic phase was washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 30B (0.961g, yield 87.9%) as a yellow liquid.
LCMS m/z＝365.3[M+1]。
The second step is that: 4- (4-Methoxypyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane (30C)
4-(4-methoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecane
30B (1.367g, 1.77mmol) was dissolved in DCM (10mL), and trifluoroacetic acid (4mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain 30C trifluoroacetate salt (0.99g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝265.3[M+1]。
The third step: 3- [2- [3- [ [4- (4-methoxypyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (30D)
tert-butyl 3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
30C trifluoroacetate (0.99g, 3.75mmol) was placed in acetonitrile (25mL), and intermediate 2(1.12g, 3.75mmol) and triethylamine (1.221g, 12.07mmol) were added in that order and stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, and extraction was performed with ethyl acetate (100mL × 2), and the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to obtain 30D as a yellow liquid (1.967g, yield 100%).
LCMS m/z＝527.3[M+1]。
The fourth step: 3- [2- [3- [ [4- (4-methoxypyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (30E)
3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
30D (1.90g, 3.61mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 30E trifluoroacetate salt (1.70g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝471.4[M+1]。
The fifth step: n- (2, 2-dimethoxyethyl) -3- [2- [3- [ [4- (4-methoxypyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (30F)
N-(2，2-dimethoxyethyl)-3-[2-[3-[2-[4-(4-methoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
The trifluoroacetate salt of 30E (3.61g, 1.70mmol) was dissolved in DCM (20mL), and triethylamine (0.73g, 7.22mmol), methylaminoacetaldehyde dimethyl acetal (0.52g, 4.33mmol), HATU (2.06g, 5.42mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 30F as a yellow liquid (2.06g, 100% yield).
LCMS m/z＝572.3[M+1]。
And a sixth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-3- [2- [3- [ [4- (4-methoxypyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (30G)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methoxpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
30F (0.500g, 0.875mmol) was dissolved in THF (10mL) and TsOH. H.was added2O (0.832g, 4.37mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.267g, 0.80mmol), stirred at room temperature for 30 minutes, then sodium triacetoxyborohydride (0.56g, 2.62mmol) was added, and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 30G (0.16G, 21% yield) as a yellow solid.
LCMS m/z＝422.8[M/2+1]。
The seventh step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-methoxypyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (Compound 30)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methoxypyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
30G (0.16G, 0.19mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.061G, 0.38mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 30(0.06g, 40% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.20(dd，1H)，7.87(d，1H)，7.08(dq，5H)，6.92-6.82(m，1H)，6.52(dd，1H)，5.89(d，1H)，5.18-5.01(m，1H)，3.74(s，3H)，3.68-3.49(m，10H)，3.49-3.40(m，3H)，3.37(dd，1H)，3.02-2.89(m，3H)，2.85-2.76(m，3H)，2.72(dd，3H)，2.54-2.43(m，4H)，2.39(t，2H)，1.72(d，2H)，1.53(t，2H).
LCMS m/z＝731.2[M+1]。
Example 31: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (4-trifluoromethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 31)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610001081
The first step is as follows: 4- (4-trifluoromethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (31B)
tert-butyl 4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxy late
Tert-butyl 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylate (0.77g, 3.0mmol) was placed in DMF (20mL), and 2-bromo-3-trifluoromethylpyrimidine (31A) (0.55g, 3.0mmol) and potassium carbonate (0.83g, 6.0mmol) were added in that order, and stirred at 50 ℃ for 4 hours. Ethyl acetate (200mL), water (100mL) was added thereto, extraction was performed, the organic phase and the aqueous phase were separated, the aqueous phase was extracted with ethyl acetate (100mL × 1), the combined organic phase was washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 31B as a yellow liquid (1.2g, yield 100%).
LCMS m/z＝425.2[M+23]。
The second step is that: 4- (4-trifluoromethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane (31C)
4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecane
31B (1.61g, 4.0mmol) was dissolved in DCM (20mL), and trifluoroacetic acid (8mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain a yellow liquid of trifluoroacetate salt of 2- (4-trifluoromethylpyrimidin-2-yl) -11-oxa-2, 9-diazaspiro [5.5] undecane (31C) (1.21g, yield 100%).
LCMS m/z＝303.2[M+1]。
The third step: 3- [2- [3- [ [4- (4-trifluoromethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (31D)
tert-butyl 3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The trifluoroacetic acid salt of 31C (1.21g, 4.0mmol) was placed in acetonitrile (25mL), and intermediate 2(1.20g, 4.0mmol), triethylamine (1.221g, 12.07mmol) and water (0.5mL) were added in that order and stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 31D as a yellow liquid (1.90g, yield 84.1%).
LCMS m/z＝565.5[M+1]。
The fourth step: 3- [2- [3- [ [4- (4-trifluoromethylpyrimidin-2-yl) 1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (31E)
3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
31D (1.88g, 3.33mmol) was dissolved in DCM (15 mL). Trifluoroacetic acid (5mL) was added dropwise to the reaction at room temperature, and the mixture was stirred for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 31E trifluoroacetic acid (1.69g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝509.3[M+1]。
The fifth step: n- (2, 2-dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (4-trifluoromethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -propionamide (31F)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
The trifluoroacetate salt of 31E (1.69g, 3.33mmol) was dissolved in DCM (20mL), and triethylamine (0.674g, 6.66mmol), methylaminoacetaldehyde dimethyl acetal (0.480g, 4.00mmol), HATU (1.90g, 5.00mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 31F as a yellow liquid (2.03g, 100% yield).
LCMS m/z＝610.4[M+1]。
And a sixth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl-N-methyl-3- [2- [3- [ [4- (4-trifluoromethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (31G)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(4-trifluoromethylpyrimidin-2-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
31F (0.500g, 0.82mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.78g, 4.10mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.267g, 0.80mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.53g, 2.46mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 31G (0.15G, 20.6% yield) as a yellow solid.
LCMS m/z＝883.5[M+1]。
The seventh step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (4-trifluoromethylpyrimidin-2-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 31)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-[4-(trifluoromethyl)pyrimidin-2-yl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
31G (0.15G, 0.17mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.055G, 0.34mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 31(0.07g, 50% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.55(d，1H)，8.36(t，1H)，7.27-7.07(m，5H)，7.03-6.94(m，1H)，6.85(d，1H)，6.74-6.56(m，1H)，5.28-5.11(m，1H)，3.90-3.79(m，2H)，3.78-3.69(m，6H)，3.69-3.62(m，2H)，3.60-3.51(m，3H)，3.51-3.43(m，1H)，3.03(s，2H)，2.96(d，1H)，2.90(s，2H)，2.86-2.75(m，3H)，2.60(dd，4H)，2.50(t，2H)，1.92(d，1H)，1.83(d，2H)，1.64(dd，2H).
LCMS m/z＝384.7[M/2+1].
Example 32: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-isopropyl-propionamide (Compound 32)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-isopropyl-propanamide
Figure GPA0000261095610001111
The first step is as follows: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-isopropyl-propionamide (32A)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-isopropyl-propanamide
22A (0.81g, 1.4mmol) was added to DCM (10mL), trifluoroacetic acid (5mL) was added dropwise at 0 ℃ and reaction was carried out at room temperature for 1 hour after completion of dropwise addition, and the system was concentrated under reduced pressure, and after addition of toluene (20mL), concentration was carried out again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.70g, 7.0mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) propan-2-amine (prepared with reference to CN 102625808) (0.31g, 2.1mmol) and HATU (0.79g, 2.1mmol) were added and allowed to warm to room temperature for 1 hour. After completion of the reaction, water (30mL) was added, DCM (30mL × 2) was extracted, the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 32A as a yellow liquid (0.85g, yield: 96%).
LCMS m/z＝640.4[M+1]
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-isopropyl-propionamide (32B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-isopropyl-propanamide
32A (0.85g, 1.3mmol) was dissolved in THF (15mL), and TsOH. H.was added2O (0.97g, 5.1mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.65g, 3.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 32B as a yellow solid (0.31g, 33% yield).
LCMS m/z＝456.9[M/2+1]。
The third step: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-isopropyl-propionamide (Compound 32)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-isopropyl-propanamide
32B (0.32g, 0.35mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.99g, 6.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 32(0.13g, 48% yield) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.23(br，1H)，8.19(dd，1H)，7.81(d，1H)，7.18(d，1H)，7.13-7.03(m，4H)，6.91(d，1H)，6.55(s，1H)，6.48(d，1H)，5.00(dd，1H)，4.04-3.99(m，2H)，3.96-3.86(m，2H)，3.66-3.49(m，9H)，3.43-3.22(d，8H)，3.19-3.13(m，2H)，2.80-2.67(m，4H)，2.65-2.57(m，2H)，2.39-2.25(m，4H)，1.67(d，2H)，1.56-1.44(s，2H)，1.08-1.06(m，2H)，1.02(d，2H)，0.57-0.49(m，2H)，0.40-0.32(m，2H).
LCMS m/z＝399.8[M/2+1].
Example 33: 3- [2- [ 2-chloro-3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (compound 33)
3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methy1-propanamide
Figure GPA0000261095610001121
The first step is as follows: [9- [ [ 2-chloro-3- (2-hydroxyethyl) phenyl ] methyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-4-yl ] - [1- (cyclopropylmethyl) pyrazol-3-yl ] methanone (33A)
[9-[[2-chloro-3-(2-hydroxyethyl)phenyl]methyl]-1-oxa-4，9-diazaspiro[5.5]undecan-4-yl]-[1-(cyclopropylmethyl)pyrazol-3-yl]methanone
1E (1.48g, 4.86mmol) was dissolved in THF (30mL), 2-chloro-3- (2-hydroxyethyl) benzaldehyde (0.9g, 5.0mmol) was added, and after stirring at room temperature for 1 hour, sodium triacetoxyborohydride (2.85g, 13.4mmol) was added, and the mixture was stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, the mixture was extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and separated by column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 33A as a yellow liquid (1.3g, 57% yield).
The second step is that: 3- [2- [ 2-chloro-3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (33B)
tert-butyl 3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
33A (1.3g, 2.7mmol) was dissolved in acetonitrile (30mL), and tert-butyl acrylate (1.38g, 10.0mmol), benzyltrimethylammonium hydroxide (0.50g, 1.2mmol, 40% solution in methanol) were added in that order and stirred at room temperature for 3 hours. Most of the reaction solvent was removed by concentration under reduced pressure, and column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10: 1) gave 33B as a yellow liquid (1.5g, yield 91%).
The third step: 3- [2- [ 2-chloro-3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (33C)
3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-methyl-propanamide
After 33B (1.5g, 2.5mmol) was added to DCM (10mL) and trifluoroacetic acid (5mL) was added dropwise at 0 ℃ and the reaction was completed at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure, and toluene (20mL) was added and concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (1.0g, 9.9mmol) were added to the residue in this order at 0 ℃ under nitrogen, and after stirring well, N- (2, 2-dimethoxyethyl) methyl-1-ammonia (0.5g, 4.2mmol) and HATU (1.8g, 4.7mmol) were added and reacted at room temperature for 1 hour. After completion of the reaction, water (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 33C as a yellow liquid (1.59g, yield 98%).
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [ 2-chloro-3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (33D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
33C (0.80g, 1.2mmol) was dissolved in THF (15mL), and TsOH. H.was added2O (1.2g, 6.2mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.33g, 1.0mmol), stirred at room temperature for 30 minutes, added sodium triacetoxyborohydride (0.79g, 3.7mmol), and stirred at room temperatureFor 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 33D as a yellow solid (0.16g, 14% yield).
The fifth step: 3- [2- [ 2-chloro-3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (compound 33)
3-[2-[2-chloro-3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]tmdecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
33D (0.16g, 0.17mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.8g, 4.9mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 33(0.08g, yield 60%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.20(br，1H)，8.20(t，1H)，7.82(d，1H)，7.36-7.28(s，1H)，7.26-7.18(d，2H)，7.08(dd，1H)，6.96(dd，1H)，6.56(s，1H)，6.49(dd，1H)，5.14-5.04(m，1H)，4.05-3.90(m，5H)，3.67-3.48(m，8H)，3.37(dd，4H)，2.93(s，3H)，2.79-2.70(m，9H)，2.44-2.32(m，4H)，1.72-1.65(m，2H)，1.58-1.48(m，2H)，0.56-0.50(m，2H)，0.40-0.34(m，2H).
LCMS m/z＝402.8[M/2+1].
Example 34: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (compound 34)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610001141
The first step is as follows: N-butyl-N- (2, 2-dimethoxyethyl) -3- [2- [3- [ [4- (4-methyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (34A)
N-butyl-N-(2，2-dimethoxyethyl)-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
23C (0.42g, 0.82mmol) was added to DCM (10mL), trifluoroacetic acid (2mL) was added dropwise at 0 ℃ and the reaction was carried out at room temperature for 1 hour, then the reaction mixture was concentrated under reduced pressure, toluene (20mL) was added and the reaction mixture was concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.40g, 3.9mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) butan-2-amine (prepared by reference to CN 102625808) (0.2g, 1.2mmol) and HATU (0.50g, 1.3mmol) were added and reacted at room temperature for 1 hour. Water (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 34A as a yellow liquid (0.42g, yield 85%).
The second step is that: N-butyl-N- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (34B)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Mixing 34A (0.20 g)0.34mmol) was dissolved in THF (15mL), and TsOH. H was added2O (0.32g, 1.7mmol), stirring at 40 ℃ for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl group]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.09g, 0.27mmol), stirred at room temperature for 30 minutes, added sodium triacetoxyborohydride (0.21g, 1.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 34B (0.11g, yield 38%) as a yellow solid
The third step: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (compound 34)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
34B (0.11g, 0.13mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.99g, 6.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 34(0.016g, 17% yield) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.24(br，1H)，8.18(d，1H)，7.93(d，1H)，7.23-7.16(m，1H)，7.14-7.04(m，4H)，6.91(d，1H)，6.65(s，1H)，6.51-6.43(m，2H)，5.10-4.87(m，1H)，3.70-3.64(m，2H)，3.61-3.54(m，4H)，3.45-3.14(m，17H)，2.79-2.60(m，6H)，2.43-2.38(m，2H)，2.32-2.24(m，2H)，2.20(s，3H)，1.87-1.64(m，3H)，1.63-1.47(m，3H).
LCMS m/z＝378.3[M/2+1]。
Example 35: N-butyl-N- [2- [2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethylamino ] ethyl ] -3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (Compound 35)
N-butyl-N-[2-[2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethylamino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610001151
34A (0.21g, 0.35mmol) was dissolved in N-methylpyrrolidone (15mL) and the acetate salt of 8- (2-aminoethyl) -5-hydroxy-4H-1, 4-benzoxazin-3-one (0.094g, 0.35mmol), acetic acid (0.021g, 0.35mmol) were added. After stirring at room temperature for 30 minutes, sodium triacetoxyborohydride (0.22g, 1.1mmol) was added and the reaction was terminated after 5 hours. Most of the reaction solvent was removed by concentration under reduced pressure, a saturated aqueous sodium bicarbonate solution (50mL) was slowly added dropwise to the residue, and extraction was performed with DCM (50mL × 2), the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, and then column chromatography separation was performed by concentration under reduced pressure [ (DCM/methanol (v/v) ═ 15: 1) ] to obtain compound 35(0.05g, yield 20%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ9.92(br，1H)，7.94(d，1H)，7.30-7.05(m，5H)，6.65(d，2H)，6.51-6.44(m，2H)，4.54-4.42(m，2H)，3.67(d，3H)，3.64-3.17(m，25H)，2.78(dd，4H)，2.67(d，1H)，2.39(d，2H)，1.77(t，2H)，1.64-1.52(m，3H)，1.51-1.37(m，2H).
LCMS m/z＝372.3[M/2+1].
Example 36: N-cyclopentyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (compound 36)
N-cyclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610001161
The first step is as follows: N-cyclopentyl-N- (2, 2-dimethoxyethyl) -3- [2- [3- [ [4- (4-methyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (36A)
N-cyclopentyl-N-(2，2-dimethoxyethyl)-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
23C (0.42g, 0.82mmol) was added to DCM (10mL), trifluoroacetic acid (2mL) was added dropwise at 0 ℃ and the reaction was carried out at room temperature for 1 hour, then the reaction mixture was concentrated under reduced pressure, toluene (20mL) was added and the reaction mixture was concentrated again under reduced pressure to give a residue. DCM (15mL) and triethylamine (0.40g, 3.9mmol) were added to the residue in this order under nitrogen at 0 ℃ and, after stirring well, N- (2, 2-dimethoxyethyl) cyclopentylamine (prepared with reference to CN 102625808) (0.20g, 0.23mmol) and HATU (0.50g, 1.3mmol) were added and reacted at room temperature for 1 hour. After completion of the reaction, water (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 36A (0.41g, yield 80%) as a yellow liquid.
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-cyclopentyl-3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (36B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-cyclopentyl-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
36A (0.21g, 0.34mmol) was dissolved in THF (15mL), and TsOH. H.was added2O (0.33g, 1.7mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.12g, 0.34mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.22g, 1.0mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 36B as a yellow solid (0.09g, 30% yield).
The third step: N-cyclopentyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (compound 36)
N-cyclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
36B (0.09g, 0.10mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.69g, 4.3mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 36(0.02g, yield 30%) as a yellow solid.
1H NMR(400MHz，DMSO-d6)δ10.24(br，1H)，8.19(d，1H)，7.93(d，1H)，7.21-7.16(m，1H)，7.14-7.05(m，4H)，6.91(d，1H)，6.65(s，1H)，6.50-6.44(m，2H)，5.03-4.96(m，1H)，3.69-3.65(m，2H)，3.61-3.54(m，4H)，3.44-3.21(m，10H)，2.77-2.73(m，2H)，2.71-2.56(dd，4H)，2.44-2.39(m，2H)，2.30-2.24(m，2H)，2.20(s，3H)，1.76-1.70(m，3H)，1.65-1.40(m，10H).
LCMS m/z＝384.2[M/2+1].
Example 37: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (Compound 37)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610001171
Figure GPA0000261095610001181
The first step is as follows: 3- [2- [3- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (37A)
tert-butyl 3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 5E (0.50g, 1.2mmol) was placed in acetonitrile (15mL), intermediate 2(0.43g, 1.2mmol), potassium carbonate (0.86g, 6.2mmol), and water (0.5mL) were added, and the mixture was stirred at room temperature for 8 hours. Water (50mL) was added, and extraction was performed with ethyl acetate (100mL × 2), and the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 37A (0.60g, yield 80%) as a yellow solid.
LCMS m/z＝568.3[M+1].
The second step is that: 3- [2- [3- [ [4- (2-propynothiazole-4-carbonyl) -11-oxa-3, 8-diazaspiro [5.5] undecan-3-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (37B)
3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoic acid
37A (0.60g, 1.0mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred for 4 h. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain a yellow liquid of trifluoroacetic acid salt of 37B (0.55g, yield 100%).
LCMS m/z＝512.3[M+1]。
The third step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [3- [ [4- (2-propynothiazole-4-carbonyl) -11-oxa-3, 8-diazaspiro [5.5] undecan-3-yl ] methyl ] phenyl ] ethoxy ] propanamide (37C)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
Trifluoroacetic acid (0.55g, 1.1mmol) of 37B was added to DCM (20mL), triethylamine (0.54g, 5.4mmol), methylaminoacetaldehyde dimethyl acetal (0.19g, 1.6mmol), HATU (0.61g, 1.6mmol) were added, and the mixture was stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 37C as a yellow solid (0.50g, 80.0% yield).
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (2-propynothiazole-4-carbonyl) -11-oxa-3, 8-diazaspiro [5.5] undecan-3-yl ] methyl ] phenyl ] ethoxy ] propionamide (37D)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
37C (0.50g, 0.8mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.80g, 4.0mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.31g, 0.92mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.5g, 2.4mmol), and stirred at room temperature for 3 hours. To this was added saturated aqueous sodium bicarbonate (50mL), extracted with DCM (100mL × 2), and the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and separated by column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give 37D as a yellow solid (0.3g, 40% yield).
LCMS m/z＝443.4[M/2+1].
The fifth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [ [4- (2-propynylthiazole-4-carbonyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (Compound 37)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[[4-(2-prop-1-ynylthiazole-4-carbonyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
37D (0.30g, 0.34mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.55g, 3.4mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 37(0.10g, yield 38%) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.03-7.89(m，2H)，7.26-7.07(m，5H)，6.76(br，1H)，6.48-6.44(m，1H)，5..0-5.27(m，1H)，3.94-3.53(m，14H)，3.10-2.59(m，14H)，2.11-1.64(m，9H).
LCMS m/z＝772.3[M+1].
Example 38: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-hetero spiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 38)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610001191
The first step is as follows: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (38A)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
22B (0.50g, 0.82mmol) was dissolved in THF (15mL), and TsOH. H.was added2O (0.78g, 4.1mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.27g, 0.82mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.35g, 1.6mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extraction was performed with DCM (100 mL. times.2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and separated by column chromatography (DCM) Methanol (v/v) at 1: 0-8: 1 gave 38A as a yellow solid (03g, 42% yield).
LCMS m/z＝885.5[M+1].
The second step is that: 3- [2- [3- [ [4- [1- (cyclopropylmethyl) pyrazole-3-carbonyl ] -1-oxa-4, 9-hetero spiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 38)
3-[2-[3-[[4-[1-(cyclopropylmethyl)pyrazole-3-carbonyl]-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
38A (0.25g, 0.28mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.46g, 2.8mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 38(0.1g, 50% yield) as a yellow solid.
1H NMR(400MHz，CDCl3)δ8.03(br，1H)，7.48-7.41(m，1H)，7.23-7.05(m，5H)，6.78-6.41(m，3H)，5.14-5.04(m，1H)，4.08-3.44(m，19H)，3.01-2.40(m，15H)，1.83-1.61(m，4H)，0.65-0.56(m，2H)，0.37-0.28(m，2H).
LCMS m/z＝771.4[M+1]。
Example 39: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-isopropyl-2-pyridinyl) -1-oxa-4, 9-oxaspirodiaza [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (Compound 39)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
Figure GPA0000261095610001201
Figure GPA0000261095610001211
The first step is as follows: 4- (4-isopropyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (39B)
tert-butyl 4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (2.56g, 10.0mmol) was dissolved in 1, 4-dioxane (50mL), and 2-chloro-4-isopropyl-pyridine (39A) (WO2008100426, 1.87g, 12.0mmol), sodium tert-butoxide (2.4g, 25.0mmol), 4, 5-bis diphenylphosphine-9, 9-dimethyloxaanthracene (0.81g, 1.4mmol), palladium acetate (0.16g, 0.7mmol), was added in order, and the reaction was warmed to reflux under nitrogen for 8 hours. After the reaction was cooled to room temperature, ethyl acetate (200mL) and water (100mL) were added, extraction was performed, the organic and aqueous phases were separated, the aqueous phase was extracted with ethyl acetate (100mL), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 39B as a yellow liquid (2.5g, 67% yield).
The second step is that: 4- (4-isopropyl-2-pyridine) -1-oxa-4, 9-diazaspiro [5.5] undecane (39C)
4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane
39B (0.75g, 2.0mmol) was weighed out, dissolved in DCM (10mL), and trifluoroacetic acid (3mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain 39C trifluoroacetate salt (0.80, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝276.1[M+1].
The third step: 3- [2- [3- [ [4- (4-isopropyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (39D)
tert-butyl 3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
After adding 39C trifluoroacetate (0.8g, 2.0mmol) to acetonitrile (15mL), tert-butyl 3- [2- [3- (2-bromomethyl) phenyl ] ethoxy ] propionate (intermediate 2) (0.71g, 2.0mmol), potassium carbonate (1.4g, 10.0mmol) and water (0.5mL) were added, and the mixture was stirred at room temperature for 8 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to obtain 39D as a yellow liquid (1.1g, yield 100%).
LCMS m/z＝539.3[M+1]。
The fourth step: 3- [2- [3- [ [4- (4-isopropyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid (39E)
3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoicacid
39D (1.1g, 2.0mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain 39E (1.0g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝482.4[M+1]。
The fifth step: n- (2, 2-Dimethoxyethyl) -3- [2- [3- [ [4- (4-isopropyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (39F)
N-(2，2-dimethoxyethyl)-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
39E (0.99g, 2.0mmol) was dissolved in DCM (20mL), and triethylamine (1.0g, 10.0mmol), methylaminoacetaldehyde dimethyl acetal (0.36g, 3.0mmol), HATU (1.2g, 3.0mmol) were added successively, followed by stirring at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and isolated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 39F as a yellow solid (0.81g, 68% yield).
LCMS m/z＝584.3[M+1]。
And a sixth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-isopropyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (39G)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
39F (0.4g, 0.7mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.7g, 3.5mmol), stirring at 40 deg.C for 1 hour, quenching the reaction by dropwise addition of saturated aqueous sodium bicarbonate (50mL), extraction with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentration under reduced pressure to give a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), and addition of 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ]Oxymethyl radical]-8-hydroxy-1H-quinolin-2-one (0.2g, 0.7mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.3g, 1.4mmol), and stirred at room temperature for 3 hours. Saturated aqueous sodium bicarbonate (50mL) was added, extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 39G (0.3G, 50% yield) as a yellow solid.
LCMS m/z＝428.4[M/2+1]。
The seventh step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-isopropyl-2-pyridinyl) -1-oxa-4, 9-oxaspirodiaza [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (Compound 39)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-isopropyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
39G (0.3G, 0.4mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.6G, 4.0mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 39(0.1g, 40% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.39-8.34(m，1H)，7.96-7.95(d，1H)，7.20-7.14(m，5H)，6.98-6.97(d，1H)，6.63-6.58(m，3H)，5.20-5.17(m，1H)，3.71-3.64(m，6H)，3.48-3.35(m，6H)，3.11(s，3H)，2.84-2.75(m，8H)，2.60-2.41(m，6H)，1.92-1.89(m，2H)，1.40-1.38(m，3H)，1.23-1.21(m，6H).
LCMS m/z＝742.5[M+1]。
Example 40: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 40)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610001231
The first step is as follows: tert-butyl 3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionate (40B)
tert-butyl 3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
3C (3.28g, 14mmol) was added to acetonitrile (50mL), and tert-butyl 3- [2- [3- (2-bromoethyl) phenyl ] ethoxy ] propionate (intermediate 1) (4.9g, 14mmol), potassium carbonate (3.87g, 28mmol), water (1mL) were added and stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 40: 1) to give 40B as a yellow liquid (4.28g, yield 60%).
The third step: N-butyl-N- (2, 2-dimethoxyethyl) -3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (40C)
N-butyl-N-(2，2-dimethoxyethyl)-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
40B (2.2g, 4.32mmol) was dissolved in DCM (15mL), and trifluoroacetic acid (10mL) was added dropwise and stirred at room temperature for 4 hours. The reaction solvent was removed by concentration under reduced pressure, toluene (10mL) was added to the residue, and after concentration under reduced pressure, DCM (30mL) and triethylamine (1.98mL, 14.2mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, N- (2, 2-dimethoxyethyl) but-1-amino (0.84g, 5.2mmol) (prepared in reference WO 2011012896) and HATU (1.97g, 5.18mmol) were added and reacted at room temperature for 30 minutes. Water (30mL) was added, DCM (30mL × 2) was extracted, the organic phases were combined, washed with brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 20) to give 40C as a clear liquid (1.62g, yield 63%).
LCMS m/z＝597.3[M+1]。
The fourth step: N-butyl-N- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (40D)
N-butyl-N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
40C (0.6g, 1.0mmol) was dissolved in THF (10mL), and TsOH. H was added2O (0.96g, 5.0mmol), stirring at 40 deg.C for 30min, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (50 mL. times.2), combining the organic phases, drying over anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, adding 5- [ 2-amino-1- (tert-butyl (dimethyl) silyl) oxy-ethyl to the residue]-8-hydroxy-1H-quinolin-2-one (0.34g, 1.0mmol), methanol (10mL) and DCM (10mL) was stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (0.64g, 3.0mmol) was added and the reaction was continued for 3 hours. Saturated sodium bicarbonate solution (30mL) was added, DCM (30mL × 2) was extracted, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 12) to give 40D as a yellow solid (0.46g, 53% yield).
LCMS m/z＝435.4[M/2+1].
The fifth step: N-butyl-N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [2- [4- (2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 40)
N-butyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[3-[2-[4-(2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
40D (0.46g, 0.53mmol) was dissolved in THF (10mL), triethylamine trihydrofluoride salt (0.41mL, 2.52mmol) was added, and the reaction was allowed to proceed at room temperature for 16 hours. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate solution (20mL), extracted with DCM (20mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 10) to give compound 40 as a yellow solid (0.21g, yield 53%).
1H NMR(400MHz，DMSO-d6)δ10.26(s，1H)，8.18(d，1H)，8.09(d，1H)，7.52(s，1H)，7.15(s，1H)，7.05(s，3H)，6.92(d，1H)，6.83(d，1H)，6.62(s，1H)，6.48(d，1H)，5.01(s，1H)，3.74-3.66(m，2H)，3.66-3.59(m，2H)，3.56(m，2H)，3.49-3.41(m，3H)，3.36-3.23(m，6H)，3.19(s，2H)，2.79-2.59(m，6H)，2.43-2.28(m，2H)，2.00(s，1H)，1.91(s，2H)，1.73(s，2H)，1.55(s，2H)，1.46(s，1H)，1.36(s，1H)，1.32-1.16(m，8H).
LCMS m/z＝378.3[M/2+1].
Example 41: 3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2-8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 41)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
Figure GPA0000261095610001241
Figure GPA0000261095610001251
The first step is as follows: 2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethanol (41B)
2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethanol
16D (1.3g, 5mmol) was dissolved in THF (30mL), 2-chloro-3- (2-hydroxyethyl) benzaldehyde (prepared by reference to CN 201080043822) (0.92g, 5mmol) was added, after stirring at room temperature for 1 hour, sodium triacetoxyborohydride (3.2g, 15mmol) was added, stirring at room temperature for 3 hours, a saturated sodium bicarbonate solution (50mL) was added to quench the reaction, DCM (60 mL. times.2) was extracted, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 40: 1) to give 41B as a brown oil (1.27g, 59% yield).
The second step is that: 3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanoic acid tert-butyl ester (41C)
tert-butyl 3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanoate
41B (1.27g, 2.95mmol) was dissolved in acetonitrile (1mL), tert-butyl acrylate (1.54g, 12mmol) was added and stirred well, then benzyltrimethylammonium hydroxide (0.17g, 1mmol) (40% in MeOH) was added and stirred at room temperature for 1 h. After completion of the reaction, the system was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 60: 1), whereby 41C was obtained as a brown-yellow oil (1.1g, yield 67%).
The third step: 3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] N- (2, 2-dimethoxyethyl) -N-methyl-propionamide (41D)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-(2，2-dimethoxyethyl)-N-methyl-propanamide
After adding DCM (20mL) to 41C (0.56g, 1mmol), trifluoroacetic acid (8mL) was added dropwise at 0 ℃ and the reaction was completed at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure, and toluene (20mL) was added and concentrated under reduced pressure. DCM (20mL) and triethylamine (0.4g, 4mmol) were added to the residue in this order under nitrogen at 0 ℃ and after stirring well, methylaminoacetaldehyde dimethyl acetal (0.14g, 1.2mmol) and HATU (0.46g, 1.2mmol) were added and reacted at room temperature for 30 minutes. After completion of the reaction, water (30mL) was added, DCM (30mL × 2) was extracted, the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 80) to give 41D as a colorless liquid (0.53g, yield 90%).
The fourth step: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (41E)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
41D (0.53g, 0.88mmol) was dissolved in THF (20mL), and TsOH. H was added2O (0.95g, 5mmol), reacted at 40 ℃ for 30 minutes. Cooling to room temperature, adding an aqueous solution (10mL) of sodium bicarbonate (0.67g, 8mmol), stirring for 10 minutes, adding a saturated solution of sodium bicarbonate (20mL), extracting with ethyl acetate (30 mL. times.2), combining the organic phases, drying over anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, adding 5- [ 2-amino-1- (tert-butyl (dimethyl) silyl) oxy-ethyl to the residue]-8-hydroxy-1H-quinolin-2-one (0.3g, 0.9mmol), methanol (10mL) and DCM (10mL) was stirred at room temperature for 1 hour, and then sodium triacetoxyborohydride (0.64g, 3mmol) was added and the reaction was continued for 3 hours. The reaction was quenched by addition of saturated sodium bicarbonate solution (30mL), extracted with DCM (30mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 12) to give 41E as a yellow solid (0.24g, 31% yield).
The fifth step: 3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2-8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide (Compound 41)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-propanamide
41E (0.24g, 0.27mmol) was dissolved in THF (20mL), triethylamine trihydrofluoride salt (0.3mL) was added, and the reaction was carried out at room temperature for 16 hours. The reaction was quenched by addition of saturated sodium bicarbonate solution (20mL), extracted with DCM (20mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 10) to give compound 41 as a light yellow solid (0.09g, 50% yield).
LCMS m/z＝381.3[M/2+1]。
1H NMR(400MHz，DMSO-d6)δ10.26(s，1H)，8.17(dd，1H)，7.97(d，1H)，7.33(s，1H)，7.31-7.15(m，2H)，7.15-7.02(m，1H)，6.92(dd，1H)，6.67(s，1H)，6.53-6.46(m，2H)，5.30(s，1H)，5.02(d，1H)，3.73-3.67(m，2H)，3.60(m，5H)，3.47-3.42(m，2H)，3.37(m，2H)，2.94(s，3H)，2.77(s，1H)，2.72(m，4H)，2.39(m，2H)，2.05-1.91(m，1H)，1.76(m，2H)，1.57(s，2H)，1.24(m，7H)，1.15(m，3H)，1.00(m，3H)，0.85(s，1H).
Example 42: 3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-cyclopentyl-N- [2- [ [ (2R) -2-hydroxy-2-8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -propionamide bistrifluoroacetate (Compound 42)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-cyclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
Figure GPA0000261095610001261
The first step is as follows: 3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] N-cyclopentyl-N- (2, 2-dimethoxyethyl) propanamide (42A)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-cyclopentyl-N-(2，2-dimethoxyethyl)propanamide
41C (0.56g, 1mmol) was added to DCM (20mL), trifluoroacetic acid (8mL) was added dropwise at 0 ℃ and after completion of the addition, the reaction mixture was concentrated under reduced pressure after 1 hour of reaction at room temperature, and toluene (20mL) was added and concentrated under reduced pressure. DCM (20mL) and triethylamine (0.4g, 4mmol) were added to the residue in this order under nitrogen at 0 ℃ and, after stirring well, cyclopentylaminoacetaldehyde dimethyl acetal (0.25g, 1.4mmol) and HATU (0.46g, 1.2mmol) were added and reacted at room temperature for 30 minutes. Water (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 80) to give 42A as a transparent liquid (0.52g, yield 80%).
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-cyclopentyl-propionamide (42B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-cyclopentyl-propanamide
42A (0.52g, 0.8mmol) was dissolved in THF (20mL) and TsOH. H was added2O (0.76g, 4mmol), reacted at 40 ℃ for 30 minutes. Cooling to room temperature, adding saturated sodium bicarbonate solution (20mL), extracting with ethyl acetate (30 mL. times.2), combining the organic phases, drying over anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, adding 5- [ 2-amino-1- (tert-butyl (dimethyl) silyl) oxy-ethyl to the residue ]-8-hydroxy-1H-quinolin-2-one (0.27g, 0.8mmol), methanol (10mL) and DCM (10mL) was stirred at room temperature for 1 hour, and then sodium triacetoxyborohydride (0.64g, 3mmol) was added and the reaction was continued for 3 hours. After the reaction was completed, the reaction was quenched with a saturated sodium bicarbonate solution (30mL), extracted with DCM (30mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 12) to give 42B as a yellow solid (0.23g, yield 31%).
The third step: 3- [2- [ 2-chloro-3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-cyclopentyl-N- [2- [ [ (2R) -2-hydroxy-2-8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -propionamide bistrifluoroacetate (Compound 42)
3-[2-[2-chloro-3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-c yclopentyl-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
42B (0.23g, 0.25mmol) was dissolved in THF (20mL), triethylamine trihydrofluoride salt (0.3mL) was added, and the reaction was carried out at room temperature for 16 hours. The reaction was quenched by addition of saturated sodium bicarbonate solution (20mL), extracted with DCM (20mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 10) and then separated and purified by a liquid phase preparative column (liquid phase preparative conditions: C18 reverse phase preparative column, mobile phases deionized water (a) containing 0.1% trifluoroacetic acid, acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution, B content 5% to 50%, elution time 15min, flow rate 12mL/min, column temperature: 30 ℃) to give compound 42(0.04g, yield 20%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ10.48(s，2H)，9.61(s，1H)，8.63(s，2H)，8.18(d，1H)，7.98(d，1H)，7.36(s，3H)，7.14(d，1H)，6.98(d，1H)，6.91(s，1H)，6.70(s，1H)，6.58(d，1H)，6.16(d，2H)，5.39-5.24(m，2H)，4.41(s，3H)，4.30(s，3H)，4.17(s，3H)，3.77(s，2H)，3.63(s，3H)，3.52(s，2H)，3.46(s，3H)，3.23(s，2H)，3.06(s，4H)，2.82(s，2H)，2.59(s，3H)，2.11(s，2H)，2.00(s，1H)，1.79(s，1H)，1.78-1.61(m，3H)，1.49(s，3H)，1.24(s，3H).
LCMS m/z＝815.3[M+1]。
Example 43: 3- [2- [3- [ [4- (4-Ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide bistrifluoroacetate (Compound 43)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide ditrifluoroacetic acid
Figure GPA0000261095610001281
The first step is as follows: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N-methyl-propionamide (43A)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-methyl-propanamide
18C (0.52g, 0.91mmol) was dissolved in THF (20mL) and TsOH. H was added2O (0.95g, 5mmol), reacted at 40 ℃ for 30 minutes, cooled to room temperature, added with saturated sodium bicarbonate solution (20mL), extracted with ethyl acetate (30 mL. times.2), combined organic phases, dried over anhydrous sodium sulfate, filtered, the filtrate concentrated under reduced pressure, and 8- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] added to the residue]Oxy-ethyl radical]-5-hydroxy-4H-1, 4-benzoxazin-3-one (0.3g, 0.9mmol), methanol (10mL) and DCM (10mL) were stirred at room temperature for 1 hour, sodium triacetoxyborohydride (0.64g, 3mmol) was added and the reaction was continued for 3 hours. After completion of the reaction, the reaction was quenched with a saturated sodium bicarbonate solution (30mL), extracted with DCM (30mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 12) to give 43A as a white solid (0.32g, yield 41%).
LCMS m/z＝845.5[M+1].
The second step is that: 3- [2- [3- [ [4- (4-Ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] ethyl ] -N-methyl-propionamide bistrifluoroacetate (Compound 43)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]ethyl]-N-methyl-propanamide ditrifluoroacetic acid
43A (0.32g, 0.38mmol) was dissolved in THF (20mL), triethylamine trihydrofluoride salt (0.4mL) was added, the reaction was carried out at room temperature for 16 hours, a saturated sodium bicarbonate solution (20mL) was added to quench the reaction, DCM (20mL × 2) was used for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 10), and then separated and purified by a liquid phase preparative column (liquid phase preparative conditions: C18 reverse phase preparative column, mobile phase was deionized water (a) containing 0.1% trifluoroacetic acid, acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution was carried out, B content was 5% to 50%, elution time was 15min, flow rate was 12mL/min, column temperature: 30 ℃) to obtain compound 43(90mg, yield 20%) as a white solid.
1H NMR(400MHz，DMSO)δ9.96(s，2H)，9.54(s，1H)，8.57(s，1H)，8.44(s，1H)，8.00(d，1H)，7.37(s，3H)，6.97-6.79(m，2H)，6.66(s，1H)，6.56(s，1H)，5.92(s，1H)，5.33(s，1H)，5.04(d，1H)，4.53(s，2H)，4.45(s，2H)，4.31(s，2H)，3.76(s，2H)，3.69-3.54(m，5H)，3.50(s，2H)，3.44(s，2H)，3.23(s，2H)，3.07(s，3H)，2.95(s，3H)，2.81(s，2H)，2.56(s，3H)，2.11(d，2H)，2.00(s，1H)，1.77-1.62(m，2H)，1.45(s，1H)，1.24(s，4H).
LCMS m/z＝366.3[M/2+1]。
Example 44: 3- [2- [3- [ [4- (4-Ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethylamino ] ethyl ] -N-methyl-propionamide bistrifluoroacetate (Compound 44)
3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[2-(5-hy droxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethylamino]ethyl]-N-methyl-propanamide ditrifluoroacetic acid
Figure GPA0000261095610001291
Dissolving 18C (0.52g, 0.91mmol) in waterTsOH. H was added to THF (20mL)2O (0.95g, 5mmol), reaction at 40 ℃ for 30 minutes, cooling to room temperature, addition of saturated sodium bicarbonate solution (20mL), extraction with ethyl acetate (30mL × 2), combination of the organic phases, drying over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, addition of the acetate salt of 8- (2-aminoethyl) -5-hydroxy-4H-1, 4-benzoxazin-3-one (0.27g, 1mmol), methanol (10mL), N-methylpyrrolidone (10mL) and acetic acid (0.06g, 1mmol) to the residue, stirring at room temperature for 1 hour, addition of sodium triacetoxyborohydride (0.64g, 3mmol), and reaction for 3 hours. Saturated sodium bicarbonate solution (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 14), followed by separation and purification with a liquid phase preparative column (liquid phase preparative condition: C18 reverse phase preparative column, mobile phase was deionized water (a) containing 0.1% trifluoroacetic acid, acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution, B content of 5% to 50%, elution time 15min, flow rate 12mL/min, column temperature: 30 ℃) to give compound 44(0.12g, yield 12%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ9.93(s，1H)，9.85(s，1H)，9.53(s，1H)，8.53(s，1H)，8.37(s，1H)，8.00(d，1H)，7.36(m，3H)，6.86(s，1H)，6.74-6.57(m，2H)，6.48(m，1H)，5.37-5.28(m，1H)，4.54(s，2H)，4.40(s，1H)，4.31(s，2H)，3.77(s，2H)，3.68-3.58(m，3H)，3.52(m，3H)，3.44(s，1H)，3.23(d，2H)，3.06(s，5H)，2.95(s，2H)，2.82(m，3H)，2.56(m，2H)，2.12(d，2H)，2.00(m，2H)，1.84(s，1H)，1.75-1.62(m，2H)，1.47(m，1H)，1.24(s，4H).
LCMS m/z＝715.4[M+1]。
Example 45: n- (cyclopropylmethyl) -3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide bistrifluoroacetate (Compound 45)
N-(cyclopropylmethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
Figure GPA0000261095610001301
The first step is as follows: n- (cyclopropylmethyl) -N- (2, 2-dimethoxyethyl) -3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (45A)
N-(cyclopropylmethyl)-N-(2，2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
After 18A (0.52g, 1mmol) was added to DCM (20mL), trifluoroacetic acid (8mL) was added dropwise at 0 ℃ and the reaction was carried out at room temperature for 1 hour, the reaction mixture was concentrated under reduced pressure, and toluene (20mL) was added and concentrated under reduced pressure. DCM (20mL) and triethylamine (0.4g, 4mmol) were added to the residue in this order under nitrogen at 0 ℃ and, after stirring well, cyclopropylaminoacetaldehyde dimethyl acetal (0.18g, 1.1mmol) and HATU (0.46g, 1.2mmol) were added and reacted at room temperature for 30 minutes. Water (30mL) was added, extraction was performed with DCM (30mL × 2), the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 60) to give 45A as a colorless liquid (0.51g, yield 83%).
LCMS m/z＝609.4[M+1]。
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N- (cyclopropylmethyl) -3- [2- [3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (45B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-(cyclopropylmethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
45A (0.51g, 0.84mmol) was dissolvedTo THF (20mL) was added TsOH. H2O (0.8g, 4.2mmol), reaction at 40 ℃ for 30 minutes, cooling to room temperature, addition of saturated sodium bicarbonate solution (20mL), extraction with ethyl acetate (30 mL. times.2), combination of the organic phases, drying over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, addition of 5- [ 2-amino-1- (tert-butyl (dimethyl) silyl) oxy-ethyl to the residue]-8-hydroxy-1H-quinolin-2-one (0.28g, 0.84mmol), methanol (10mL) and DCM (10mL) was stirred at room temperature for 1 hour, and then sodium triacetoxyborohydride (0.64g, 3mmol) was added and the reaction was continued for 3 hours. After the reaction was completed, the reaction was quenched with a saturated sodium bicarbonate solution (30mL), extracted with DCM (30mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 12) to give 45B as a yellow solid (0.29g, yield 39%).
LCMS m/z＝441.4[M/2+1]
The third step: n- (cyclopropylmethyl) -3- [2- [3- [ [4- (4-ethyl-2-piperidinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide bistrifluoroacetate (Compound 45)
N-(cyclopropylmethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
45B (0.29g, 0.33mmol) was dissolved in THF (20mL), triethylamine trihydrofluoride salt (0.3mL) was added, and the reaction was carried out at room temperature for 16 hours. Saturated sodium bicarbonate solution (20mL) was added, DCM (20mL × 2) was extracted, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 10) and then separated and purified by a liquid phase preparative column (liquid phase preparative conditions: C18 reverse phase preparative column, mobile phase was deionized water (a) containing 0.1% trifluoroacetic acid, acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution, B content ═ 5% to 50%, elution time 15min, flow rate 12mL/min, column temperature: 30 ℃) to give compound 45 as a white solid (0.06g, yield 20%).
1H NMR(400MHz，DMSO-d6)δ10.49(s，2H)，9.55(s，1H)，8.71(s，1H)，8.57(s，1H)，8.16(m，1H)，7.99(m，1H)，7.36(s，3H)，7.15(m，1H)，7.03-6.93(m，1H)，6.85(s，1H)，6.60(m，2H)，6.17(s，2H)，5.43-5.22(m，2H)，4.30(s，3H)，3.76(s，4H)，3.64(m，6H)，3.44(s，3H)，3.14(m，7H)，2.82(m，2H)，2.71-2.55(m，3H)，2.11(m，2H)，2.00(m，2H)，1.83(s，1H)，1.69(m，2H)，1.40(s，1H)，1.24(s，5H).
LCMS m/z＝384.4[M/2+1].
Example 46: n-cyclopentyl-3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide bistrifluoroacetate (Compound 46)
N-cyclopentyl-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
Figure GPA0000261095610001311
Figure GPA0000261095610001321
The first step is as follows: N-cyclopentyl-N- (2, 2-dimethoxyethyl) -3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propanamide (46A)
N-cyclopentyl-N-(2，2-dimethoxyethyl)-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
After 18A (0.52g, 1mmol) was added to DCM (20mL), trifluoroacetic acid (8mL) was added dropwise at 0 ℃ and the reaction was completed at room temperature for 1 hour, the system was concentrated under reduced pressure, and toluene (20mL) was added and concentrated under reduced pressure. DCM (20mL) and triethylamine (0.4g, 4mmol) were added to the residue in this order under nitrogen at 0 ℃ and, after stirring well, cyclopentylaminoacetaldehyde dimethyl acetal (0.19g, 1.1mmol) and HATU (0.46g, 1.2mmol) were added and reacted at room temperature for 30 minutes. Water (30mL) was added, DCM (30mL × 2) was extracted, the organic phases were combined, washed with saturated brine (40mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 60) to give 46A as a colorless liquid (0.56g, yield 89%).
LCMS m/z＝623.4[M+1]
The second step is that: n- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-cyclopentyl-3- [2- [3- [ [4- (4-ethyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] propionamide (46B)
N-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-cyclopentyl-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]propanamide
46A (0.56g, 0.9mmol) was dissolved in THF (20mL) and TsOH. H was added2O (0.95g, 5mmol), reaction at 40 ℃ for 30 minutes, cooling to room temperature, addition of saturated sodium bicarbonate solution (30mL), extraction with ethyl acetate (30 mL. times.2), combination of the organic phases, drying over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, addition of 5- [ 2-amino-1- (tert-butyl (dimethyl) silyl) oxy-ethyl to the residue]-8-hydroxy-1H-quinolin-2-one (0.3g, 0.9mmol), methanol (10mL) and DCM (10mL) was stirred at room temperature for 1 hour, and then sodium triacetoxyborohydride (0.64g, 3mmol) was added and the reaction was continued for 3 hours. After completion of the reaction, the reaction was quenched with saturated sodium bicarbonate solution (30mL), extracted with DCM (30mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 12) to give 46B as a yellow solid (0.35g, yield: 43%).
LCMS m/z＝448.4[M/2+1]。
The third step: n-cyclopentyl-3- [2- [3- [ [4- (4-ethyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethoxy ] -N- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] propionamide bistrifluoroacetate (Compound 46)
N-cyclopentyl-3-[2-[3-[[4-(4-ethyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethoxy]-N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]propanamide ditrifluoroacetic acid
46B (0.35g, 0.39mmol) was dissolved in THF (20mL), triethylamine trihydrofluoride salt (0.4mL) was added, and the reaction was carried out at room temperature for 16 hours. Saturated sodium bicarbonate solution (20mL) was added, DCM (20mL × 2) was extracted, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/methanol (v/v) ═ 1: 10) and then separated and purified by a liquid phase preparative column (liquid phase preparative conditions: C18 reverse phase preparative column, mobile phase was deionized water (a) containing 0.1% trifluoroacetic acid, acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution, B content ═ 5% to 50%, elution time 15min, flow rate 12mL/min, column temperature: 30 ℃) to give compound 46(0.11g, yield: 28%) as a white solid.
1H NMR(400MHz，DMSO-d6)δ10.48(s，2H)，9.61(s，1H)，8.63(s，2H)，8.18(d，1H)，7.98(d，1H)，7.36(s，3H)，7.14(d，1H)，6.98(d，1H)，6.91(s，1H)，6.70(s，1H)，6.58(d，1H)，6.29-6.04(m，1H)，5.39-5.22(m，2H)，4.41(s，1H)，4.30(s，2H)，4.17(s，1H)，3.77(s，2H)，3.63(d，4H)，3.52(s，2H)，3.46(s，3H)，3.35-2.94(m，7H)，2.82(s，2H)，2.59(s，3H)，2.33(s，1H)，2.11(d，2H)，2.02-1.89(m，1H)，1.79(s，1H)，1.74-1.60(m，3H)，1.49(s，4H)，1.24(s，3H).
LCMS m/z＝391.3[M/2+1]。
Example 47: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-oxo-1H-pyridin-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 47)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
Figure GPA0000261095610001331
The first step is as follows: 4- (2-benzyloxy-3-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (47B)
tert-butyl 4-(2-benzyloxy-3-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (0.512g, 2.00mmol) was added to toluene (50mL), 2-benzyloxy-3-bromopyridine (47A) (0.528g, 2.00mmol), sodium tert-butoxide (0.40g, 4.16mmol), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.036g, 0.062mmol), tris (dibenzylideneacetone) dipalladium (0.036g, 0.039mmol) were added, and the reaction was carried out at 100 ℃ under nitrogen for 6 hours. The reaction mixture was cooled to room temperature, ethyl acetate (200mL) and water (100mL) were added thereto, extraction was performed, the organic and aqueous phases were separated, the aqueous phase was extracted with ethyl acetate (100mL), the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 47B (0.812g, 93% yield) as a yellow liquid.
LCMS m/z＝440.2[M+1]。
The second step is that: 4- (2-oxo-1H-pyridin-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (47C)
tert-butyl 4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecane-9-carboxylate
47B (0.89g, 2.02mmol) was added to ethyl acetate (20mL), palladium on carbon (0.08g, 10% w/w) was added, and the reaction was carried out for 4 hours under a hydrogen atmosphere. The reaction mixture was filtered through celite, the filter cake was washed with ethyl acetate (100ml), the filtrate was concentrated under reduced pressure and subjected to column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1: 1) to give 47C as a yellow liquid (0.700g, yield 99%).
LCMS m/z＝372.2[M+23]。
The third step: 3- (1-oxa-4, 9-diazaspiro [5.5] undecan-4-yl) -1H-pyridin-2-one (47D)
3-(1-oxa-4，9-diazaspiro[5.5]undecan-4-yl)-1H-pyridin-2-one
47C (0.70g, 2.00mmol) was dissolved in DCM (20mL), and trifluoroacetic acid (4mL) was added and stirred at room temperature for 2 hours. The reaction solvent was removed by concentration under reduced pressure, and toluene (5mL) was added to the residue to obtain 47D trifluoroacetate salt (0.50g, yield 100%) as a yellow liquid after concentration under reduced pressure.
LCMS m/z＝250.2[M+1].
The fourth step: 3- [2- [3- [2- [4- (2-oxo-1H-pyridin-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionic acid tert-butyl ester (47E)
tert-butyl 3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoate
The trifluoroacetate salt of 47D (0.500g, 2.00mmol) was placed in acetonitrile (25mL) and intermediate 1(0.720g, 2.00mmol), potassium carbonate (1.589g, 11.50mmol), water (0.5mL) were added. Stirred at 60 ℃ for 24 hours. After the reaction was cooled to room temperature, water (50mL) was added, extraction was performed with ethyl acetate (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 47E (0.43g, yield 41%) as a yellow liquid.
LCMS m/z＝526.3[M+1]。
The fifth step: 3- [2- [3- [2- [4- (2-oxo-1-H-pyridin-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propanoic acid (47F)
3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanoic acid
47E (0.43, 0.818mmol) was added to DCM (15mL), and trifluoroacetic acid (5mL) was added dropwise and stirred at room temperature for 4 h. The reaction solvent was removed by concentration under reduced pressure, and toluene (10mL) was added to the residue to obtain the yellow liquid trifluoroacetate salt of 47F (0.38, yield 99%).
LCMS m/z＝470.3[M+1]。
And a sixth step: n- (2, 2-Dimethoxyethyl) -N-methyl-3- [2- [3- [2- [4- (2-oxo-1H-pyridin-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (47G)
N-(2，2-dimethoxyethyl)-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
The trifluoroacetic acid salt of 47F (0.38g, 0.81mmol) was dissolved in DCM (20mL), triethylamine (0.246g, 2.43mmol), methylaminoacetaldehyde dimethyl acetal (0.116g, 0.973mmol), HATU (0.462g, 1.21mmol) were added, and the mixture was stirred at room temperature for 2 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise, extracted with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (50mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 47G (0.46G, 99% yield) as a yellow liquid.
LCMS m/z＝571.4[M+1]。
The seventh step: n- [2- [ [ (2R) -2- (tert-butyl (dimethyl) silyl) oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-oxo-1H-pyridin-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (47H)
N-[2-[[(2R)-2-(tert-butyl(dimethyl)silyl)oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
47G (0.465G, 0.81mmol) was dissolved in THF (10mL), and TsOH. H.was added2O (0.77g, 4.04mmol), stirring at 40 deg.C for 1 hr, quenching the reaction by dropwise adding saturated aqueous sodium bicarbonate (50mL), extracting with DCM (100 mL. times.2), drying the combined organic phases over anhydrous sodium sulfate, concentrating under reduced pressure to obtain a crude product, dissolving in a mixed solvent of DCM (10mL) and methanol (3mL), adding 5- [ 2-amino-1- (tert-butyl (dimethyl) silyl) oxy-ethyl]-8-hydroxy-1H-quinolin-2-one (0.27g, 0.80mmol), stirred at room temperature for 30 minutes, then added sodium triacetoxyborohydride (0.51g, 2.42mmol), and stirred at room temperature for 3 hours. Is added intoThe reaction was quenched with aqueous sodium bicarbonate (50mL), extracted with DCM (100mL × 2), the combined organic phases dried over anhydrous sodium sulfate, concentrated under reduced pressure and isolated by column chromatography (DCM: methanol (v/v) ═ 1: 0-8: 1) to give 47H as a yellow solid (0.40g, 59% yield).
LCMS m/z＝422.4[M/2+1]。
Eighth step: n- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethyl ] -N-methyl-3- [2- [3- [2- [4- (2-oxo-1H-pyridin-3-yl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] ethyl ] phenyl ] ethoxy ] propionamide (Compound 47)
N-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl]-N-methyl-3-[2-[3-[2-[4-(2-oxo-1H-pyridin-3-yl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]ethyl]phenyl]ethoxy]propanamide
47H (0.40g, 0.474mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.22g, 1.1mmol) was added, and the mixture was stirred at room temperature for 12 hours. Saturated aqueous sodium bicarbonate (50mL) was added dropwise in this order, the reaction was quenched with methanol (5mL), extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 47(0.076g, 22% yield) as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.37(dd，1H)，7.28-7.14(m，2H)，7.13-6.96(m，5H)，6.91(d，1H)，6.63(t，1H)，6.34(t，1H)，5.24(dt，1H)，3.93-3.79(m，2H)，3.73(dd，2H)，3.69-3.42(m，5H)，3.13-2.74(m，20H)，2.63(t，2H)，2.20(d，2H)，1.85(t，2H).
LCMS m/z＝730.4[M+1].
Example 48: compounds 48-138 of the present invention, compounds (which can be summarized by formula I) are synthesized as follows:
Figure GPA0000261095610001361
step 1): compound a (LG ═ Cl, Br, I, or other leaving group) (1.5eq.) is added to a solvent (e.g., 1, 4-dioxane, DMF, acetonitrile) with 1-oxa-4, 9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester (1.0eq.) under conditions of 1 (sodium tert-butoxide (2.0eq.), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (0.15eq.) and palladium acetate (0.07eq.), reflux under nitrogen atmosphere) or 2 (potassium carbonate (2.0eq.) or triethylamine (3.0eq.), 40 ℃ to reflux of the reaction), and the reaction temperature may be increased depending on the reaction monitoring, until the end of the reaction monitored by TLC or LC-MS. Obtaining a compound B by adopting conventional post-treatment such as extraction and column chromatography separation;
step 2): compound B was dissolved in DCM and trifluoroacetic acid (1/4 volume DCM) was added and the reaction was monitored by TLC or LC-MS at rt to completion. Directly concentrating under reduced pressure to obtain trifluoroacetate of compound C (directly used for next reaction), or performing conventional post-treatment such as adding water, adjusting pH value of the solution to neutrality, extracting, and performing column chromatography separation to obtain compound C.
And 3) dissolving the compound C or the trifluoroacetate (1.0eq) thereof in acetonitrile, adding 1.0eq of any one of the intermediates 1-6, 5.0eq of potassium carbonate and 0.5mL of water, and reacting at 40-60 ℃ until the reaction is finished by TLC or LC-MS monitoring. And separating by conventional post-treatment such as extraction and column chromatography to obtain compound D.
Step 4) Compound D was dissolved in DCM and trifluoroacetic acid (1/3 volume of DCM) was added and the reaction was stopped by TLC or LC-MS monitoring at RT. Directly concentrating under reduced pressure to obtain compound E (directly used for next reaction), or performing conventional post-treatment such as adding water, adjusting pH value of the solution to neutrality, extracting, and separating by column chromatography to obtain compound E.
Step 5) Compound E (1.0eq) is dissolved in DCM and Compound F (R) is addedxMethyl, ethyl or cyclopentyl) (1.5eq), triethylamine (3-5 eq.), and HATU (1.5eq.) were reacted at room temperature until the end of the reaction was monitored by TLC or LC-MS. Adding saturated sodium bicarbonate water solution to quench the reaction, extracting and separating by column chromatography to obtain a compound G.
Step 6) Compound G (1.0eq.) was dissolved in THF, and TsOH. H.was added thereto2O (5.0eq), 40 ℃ to TLC or LC-MS to monitor the end of the reaction. Adding saturated sodium bicarbonate water solution to quench reaction, extracting, and reducing pressureThe concentrated product was dissolved in a mixed solvent of DCM and methanol (v/v ═ 10/3), compound H (1.0eq) was added, and after stirring at room temperature for 30min, sodium triacetoxyborohydride (3.0eq.) was added to react at room temperature until the reaction was completed, monitored by TLC or LC-MS. Adding saturated sodium bicarbonate water solution to quench reaction, extracting and separating by column chromatography to obtain the product. When R in the compound H 9When H, the product obtained is compound I. When R in the compound H9When OTBS is used, the product (1.0eq.) is further dissolved in THF, triethylamine trihydrofluoric acid (1.1eq.) is added, and the reaction is carried out at room temperature until the end of the reaction is monitored by TLC or LC-MS. Adding saturated sodium bicarbonate water solution to quench reaction, extracting, and separating by column chromatography to obtain compound I.
In the synthesis of the compound I, the residue after concentration after extraction can also be separated and purified by preparative liquid chromatography, and the chromatographic conditions are as follows: c18 reverse phase preparative column, mobile phase deionized water (a) containing 0.1% trifluoroacetic acid, acetonitrile (B) containing 0.1% trifluoroacetic acid, gradient elution B: (A + B) ═ 5% to 20%, elution time 15min, flow rate 12mL/min, column temperature: 30 ℃) and the compound I obtained after separation and purification by preparative liquid chromatography is in the form of ditrifluoroacetate salt. The chromatographic conditions can be modified by those skilled in the art according to conventional means to isolate and purify the product.
Meanwhile, the compound 48 to the compound 149 (which can be summarized by the general formula I) can also be synthesized into a desired intermediate by the methods in examples 1 to 47 and intermediates 1 to 15.
Compound 48
Figure GPA0000261095610001371
1H NMR(400MHz，DMSO-d6)δ10.23(br，1H)，8.17(d，1H)，8.05(t，2H)，7.56(t，1H)，7.45(t，1H)，7.19-7.12(m，1H)，7.10-6.98(m，4H)，6.92(d，1H)，6.48(dd，1H)，5.05-4.96(m，1H)，3.89-3.78(m，2H)，3.64-3.52(m，4H)，3.40-3.36(m，2H)，3.34-3.29(m，2H)，3.26(s，2H)，2.92(s，1H)，2.78-2.64(m，9H)，2.62-2.52(m，6H)，2.36(t，2H)，1.95(d，2H)，1.66(t，2H)，1.23(s，2H).
LCMS m/z＝385.3[M/2+1].
Compound 49
Figure GPA0000261095610001372
1H NMR(400MHz，DMSO-d6)δ10.28(br，1H)，8.17(dd，1H)，7.82(d，1H)，7.18(s，1H)，7.14-7.04(m，4H)，6.94-6.88(m，1H)，6.55(s，1H)，6.48(dd，1H)，5.05-4.97(m，1H)，4.04-3.99(m，2H)，3.96-3.87(m，2H)，3.66-3.52(m，8H)，3.38-3.24(m，14H)，2.80-2.61(m，6H)，2.38-2.24(m，4H)，1.71-1.63(m，2H)，1.58-1.40(m，4H)，0.56-0.49(m，2H)，0.40-0.33(m，2H).
LCMS m/z＝406.8[M/2+1].
Compound 50
Figure GPA0000261095610001381
1H NMR(400MHz，DMSO-d6)δ10.26(br，1H)，8.18(d，1H)，7.94(d，1H)，7.22-7.04(m，6H)，6.91(d，1H)，6.65(s，1H)，6.50-6.45(m，2H)，5.05-4.95(m，1H)，3.68-3.65(m，2H)，3.63-3.53(m，5H)，3.43-3.39(s，5H)，3.35-3.20(m，12H)，2.79-2.63(m，6H)，2.43-2.39(m，2H)，2.30-2.24(m，2H)，1.76-1.71(m，2H)，1.57-1.50(m，2H).
LCMS m/z＝728.4[M+1].
Compound 51
Figure GPA0000261095610001382
1H NMR(400MHz，DMSO-d6)δ9.82(br，1H)，7.82(d，1H)，7.27-6.94(m，6H)，6.86(d，1H)，6.55(s，1H)，6.50(d，1H)，4.83(dd，1H)，4.03-3.99(m，2H)，3.94-3.88(m，2H)，3.65-3.53(m，9H)，3.44-3.24(m，12H)，2.80-2.72(m，3H)，2.70-2.60(m，3H)，2.59-2.53(m，2H)，2.35-2.25(m，4H)，1.75-1.42(m，7H)，0.57-0.52(m，2H)，0.40-0.35(m，2H).
LCMS m/z＝828.5[M+1]。
Compound 52
Figure GPA0000261095610001383
1H NMR(400MHz，Methanol-D4)δ8.01-7.92(m，2H)，7.38-7.25(m，5H)，6.95-6.93(m，1H)，6.72-6.70(d，1H)，6.51-6.49(d，1H)，3.92-3.91(m，2H)，3.74-3.54(m，10H)，3.40-3.18(m，14H)，2.90-2.87(m，4H)，2.63-2.61(m，2H)，2.26-2.22(m，2H)，1.97-1.85(m，2H).
LCMS m/z＝687.4[M+1].
Compound 53 and compound 54
Figure GPA0000261095610001384
Compound 53RxEthyl group; LCMS M/z 364.8[ M/2+1].
Compound 54RxIs cyclopropyl; LCMS M/z 370.8[ M/2+1 ]].
Compound 55
Figure GPA0000261095610001391
LCMS m/z＝342.2[M/2+1].
Compound 56
Figure GPA0000261095610001392
LCMS m/z＝370.8[M/2+1]。
Compound 57
Figure GPA0000261095610001393
1H NMR(400MHz，DMSO-d6)δ10.24(br，1H)，8.19(d，1H)，7.82(d，1H)，7.26-7.01(m，6H)，6.92(d，1H)，6.55(s，1H)，6.49(d，1H)，5.06-4.94(m，1H)，4.03-3.98(m，2H)，3.96-3.89(m，2H)，3.67-3.48(m，10H)，3.44-3.24(m，10H)，3.21-3.08(m，3H)，2.78-2.68(m，4H)，2.62(t，2H)，2.58-2.53(m，1H)，2.36-2.25(m，4H)，1.74-1.39(m，4H)，0.53(s，2H)，0.37(s，2H).
LCMS m/z＝824.5[M+1].
Compound 58
Figure GPA0000261095610001394
1H NMR(400MHz，CDCl3)δ8.62(s，1H)，7.98(d，1H)，7.60(s，1H)，7.30-7.27(m，1H)，7.24-7.12(m，3H)，6.66(d，1H)，6.56-6.40(m，4H)，4.57-4.48(m，3H)，4.09(s，3H)，3.85-3.58(m，9H)，3.50-3.42(m，3H)，3.37(s，2H)，3.27-3.16(m，6H)，2.95-2.89(m，3H)，2.86-2.78(m，3H)，2.61-2.55(m，2H)，2.13-1.95(m，6H).
LCMS m/z＝701.6[M+1].
Compound 59
Figure GPA0000261095610001395
1H NMR(400MHz，DMSO-d6)δ9.78(s，1H)，7.82(d，1H)，7.22-7.15(m，1H)，7.14-7.04(m，3H)，6.59(d，1H)，6.55(s，1H)，6.41(d，1H)，4.46(d，2H)，4.04-3.98(m，2H)，3.97-3.87(m，2H)，3.70-3.45(m，9H)，3.35-3.23(m，16H)，2.75(t，2H)，2.65-2.59(m，3H)，2.58-2.54(m，2H)，2.39-2.25(m，4H)，1.70-1.63(m，2H)，1.55-1.46(m，2H)，0.57-0.49(s，2H)，0.38(s，2H).
LCMS m/z＝400.8[M/2+1]。
Compound 60
Figure GPA0000261095610001401
1H NMR(400MHz，CD3OD)δ8.05(d，1H)，7.89(d，1H)，7.48-7.39(m，2H)，7.30(d，1H)，7.24-7.18(m，2H)，7.04(d，1H)，6.96(d，1H)，6.65(d，1H)，5.40(dd，1H)，4.40(s，2H)，3.96-3.91(m，2H)，3.81-3.57(m，11H)，3.46-3.39(m，2H)，3.35(s，2H)，3.29-3.21(m，4H)，3.09(s，3H)，2.96-2.90(m，2H)，2.76(q，2H)，2.67(t，2H)，2.24(d，2H)，2.00-1.85(m，2H)，1.28(t，3H).
LCMS m/z＝373.3[M/2+1].
Compound 61
Figure GPA0000261095610001402
1H NMR(400MHz，CDCl3)δ8.00(d，1H)，7.22-7.05(m，4H)，6.54(d，1H)，6.46(d，1H)，6.41(s，1H)，6.27-6.19(m，1H)，4.54(s，2H)，3.81-3.76(m，2H)，3.71(dd，2H)，3.61-3.55(m，2H)，3.50(s，2H)，3.48-3.44(m，3H)，3.39-3.27(m，5H)，2.88-2.68(m，8H)，2.62-2.51(m，4H)，2.43(t，2H)，1.91(d，2H)，1.68-1.60(m，2H)，1.30-1.21(m，3H)，1.11(t，2H)，1.06(t，1H).
LCMS m/z＝358.3[M/2+1].
Compound 62
Figure GPA0000261095610001403
1H NMR(400MHz，DMSO-d6)δ9.77(s，1H)，7.82(d，1H)，7.18(s，1H)，7.13-7.02(m，3H)，6.60(d，1H)，6.55(s，1H)，6.42(d，1H)，4.46(d，2H)，4.04-3.98(m，2H)，3.96-3.86(m，2H)，3.68-3.47(m，9H)，3.44-3.19(m，12H)，2.76(t，2H)，2.66-2.60(m，3H)，2.60-2.53(m，2H)，2.37-2.23(m，4H)，1.71-1.62(m，2H)，1.56-1.44(m，2H)，0.54(s，2H)，0.37(s，2H).
LCMS m/z＝386.8[M/2+1].
Compound 63
Figure GPA0000261095610001411
1H NMR(400MHz，CD3OD)δ8.43(d，1H)，7.76(s，1H)，7.52-7.40(m，2H)，7.33(d，1H)，7.25(s，1H)，7.07(d，1H)，6.68(d，2H)，5.44(d，1H)，4.42(s，2H)，4.31(dd，1H)，4.13-4.00(m，4H)，3.87-3.51(m，11H)，3.47-3.39(m，2H)，3.32-3.17(m，5H)，2.98(t，2H)，2.76(t，2H)，2.31-2.17(m，2H)，1.97-1.89(m，2H)，1.89-1.75(m，4H)，1.68-1.61(m，2H)，1.59-1.47(m，2H)，1.37-1.28(m，2H)，0.66-0.57(m，2H)，0.42(s，2H).
LCMS m/z＝421.8[M/2+1].
Compound 64
Figure GPA0000261095610001412
1H NMR(400MHz，DMSO-d6)δ9.89(dd，2H)，8.82(s，1H)，7.83(s，1H)，7.55-7.17(m，4H)，6.67(d，1H)，6.57(s，1H)，6.50(dd，1H)，4.54(s，2H)，4.30-4.16(m，1H)，4.14-4.06(m，1H)，4.01(d，3H)，3.95-3.87(m，1H)，3.71-3.56(m，7H)，3.54-3.40(m，3H)，3.30(s，5H)，3.08-3.02(m，3H)，2.99-2.92(m，2H)，2.87-2.76(m，4H)，2.60(t，2H)，2.03-1.75(m，4H)，1.35(d，2H)，1.12(d，3H)，1.05(d，1H)，0.53(d，2H)，0.37(d，2H).
LCMS m/z＝393.8[M/2+1].
Compound 65
Figure GPA0000261095610001413
1H NMR(400MHz，DMSO-d6)δ10.24(br，1H)，8.18(dd，1H)，7.82(d，1H)，7.19(s，2H)，7.06(dt，2H)，6.93(dd，1H)，6.55(s，1H)，6.49(dd，1H)，5.08-5.01(m，1H)，4.01(d，2H)，3.96-3.87(m，2H)，3.65-3.54(m，7H)，3.49(s，2H)，3.43(s，1H)，3.35-3.24(m，6H)，2.84-2.65(m，7H)，2.40-2.27(m，4H)，1.71-1.63(m，2H)，1.55-1.44(m，2H)，1.03(t，3H)，0.95(t，1H)，0.56-0.49(m，2H)，0.37(s，2H).
LCMS m/z＝802.5[M+1].
Compound 66
Figure GPA0000261095610001414
1H NMR(400MHz，DMSO-d6)δ9.81(br，1H)，7.82(d，1H)，7.22-7.17(m，1H)，7.15-7.06(m，3H)，6.86(d，1H)，6.55(s，1H)，6.50(d，1H)，4.87-4.78(m，1H)，4.52-4.42(m，2H)，4.04-3.98(m，2H)，3.96-3.88(m，2H)，3.67-3.60(m，4H)，3.59-3.44(m，5H)，3.44-3.21(m，7H)，2.79-2.74(m，2H)，2.71-2.63(m，5H)，2.62-2.54(m，4H)，2.38-2.26(m，4H)，1.71-1.64(m，2H)，1.56-1.46(m，2H)，0.54(s，2H)，0.37(s，2H).
LCMS m/z＝800.5[M+1].
Compound 67
Figure GPA0000261095610001421
1H NMR(400MHz，DMSO-d6)δ10.32(s，1H)，8.29(d，1H)，8.20(t，1H)，7.98(dd，1H)，7.32-7.27(m，1H)，7.21-7.07(m，6H)，6.95(dd，1H)，6.50(dd，1H)，5.21-5.07(m，1H)，3.77-3.71(m，2H)，3.65-3.50(m，5H)，3.47-3.41(m，3H)，3.13-3.08(m，2H)，3.03(s，2H)，2.94(s，2H)，2.91-2.82(m，4H)，2.81-2.73(m，5H)，2.58-2.52(m，2H)，2.48-2.45(m，1H)，2.29(t，2H)，1.83(d，2H)，1.60(t，2H).
LCMS m/z＝350.3[M/2+1].
Compound 68
Figure GPA0000261095610001422
1H NMR(400MHz，DMSO-d6)δ7.82(d，1H)，7.19(s，1H)，7.14-7.05(m，3H)，6.85(d，1H)，6.69(d，1H)，6.55(s，1H)，4.59-4.52(m，1H)，4.04-3.98(s，2H)，3.96-3.85(m，2H)，3.67-3.46(m，9H)，3.45-3.33(m，4H)，3.30(t，4H)，2.82-2.71(m，4H)，2.63(qd，4H)，2.39-2.25(m，4H)，1.68(d，2H)，1.56-1.46(s，2H)，0.57-0.50(m，2H)，0.38(s，2H).
LCMS m/z＝388.8[M/2+1].
Compound 69
Figure GPA0000261095610001423
1H NMR(400MHz，DMSO-d6)δ7.81(d，1H)，7.21-7.16(m，1H)，7.14-7.05(m，3H)，6.85(d，1H)，6.71-6.67(m，1H)，6.55(s，1H)，4.59-4.52(m，1H)，4.04-3.98(m，2H)，3.98-3.88(m，2H)，3.70-3.46(m，9H)，3.45-3.34(m，3H)，3.31-3.19(m，5H)，2.76(t，2H)，2.69-2.57(m，4H)，2.48-2.44(m，2H)，2.39-2.26(m，4H)，1.67(d，2H)，1.56-1.45(m，2H)，1.04(t，2H)，0.57-0.50(m，2H)，0.38(s，2H).
LCMS m/z＝395.8[M/2+1].
Compound 70
Figure GPA0000261095610001431
1H NMR(400MHz，DMSO-d6)δ10.06(br，1H)，8.37(d，1H)，8.17(d，1H)，7.47(d，1H)，7.23-7.16(m，2H)，7.13(s，1H)，7.10(d，2H)，7.06(d，1H)，6.91(d，1H)，6.48(dd，1H)，5.02-4.96(m，1H)，3.73-3.67(m，2H)，3.62-3.52(m，8H)，3.42(d，2H)，3.37-3.23(m，4H)，2.79-2.71(m，4H)，2.71-2.57(m，4H)，2.53(d，1H)，2.47-2.39(m，3H)，2.26(t，2H)，1.72(d，2H)，1.60-1.52(m，2H).
LCMS m/z＝372.8[M/2+1].
Compound 71
Figure GPA0000261095610001432
Compound 70(0.25g, 0.34mmol) was dissolved in methanol (10mL), palladium on carbon (0.1g) was added, and the reaction was stirred at room temperature for 4 hours under a hydrogen atmosphere. The reaction mixture was filtered through celite, concentrated under reduced pressure, and separated by column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 71 as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.40(d，1H)，7.54(d，1H)，7.35(dd，4H)，7.29(d，1H)，7.03(d，1H)，6.65(d，1H)，6.33(dd，1H)，6.07(s，1H)，5.38(dd，1H)，4.29(s，2H)，3.91-3.85(m，2H)，3.75(dd，3H)，3.69(t，3H)，3.50-3.31(m，7H)，3.28-3.18(m，5H)，3.08(s，3H)，2.89(t，2H)，2.66(t，2H)，2.22(d，2H)，1.86(t，2H)，1.33-1.27(m，1H).
LCMS m/z＝715.5[M+1].
Compound 72
Figure GPA0000261095610001433
1H NMR(400MHz，DMSO-d6)δ10.49(s，1H)，9.41(d，1H)，8.08(dd，1H)，7.23-7.17(m，1H)，7.12(dd，3H)，6.69(d，1H)，6.52-6.47(m，1H)，6.39(d，1H)，4.10-3.99(m，1H)，3.68-3.65(m，2H)，3.64-3.60(m，2H)，3.59-3.54(m，2H)，3.50-3.44(m，3H)，3.44-3.37(m，5H)，3.30(s，6H)，2.92(d，3H)，2.84(d，1H)，2.80-2.75(m，2H)，2.72-2.67(m，1H)，2.44-2.39(m，2H)，2.30-2.23(m，2H)，1.72(d，2H)，1.57-1.50(m，2H).
LCMS m/z＝360.3[M/2+1].
Compound 73
Figure GPA0000261095610001441
1H NMR(400MHz，DMSO-d6)δ10.15(br，1H)，8.39(dd，1H)，8.23(dd，1H)，8.17(d，1H)，7.18(dt，1H)，7.13-7.05(m，4H)，6.90(dd，2H)，6.48(dd，1H)，5.02-4.97(m，1H)，3.72-3.66(m，2H)，3.65-3.48(m，5H)，3.40(d，2H)，3.35(s，2H)，3.29(dd，3H)，3.23-3.16(m，3H)，2.79-2.73(m，4H)，2.71-2.61(m，4H)，2.53(d，1H)，2.42-2.35(m，2H)，2.28(t，2H)，1.72(d，2H)，1.56-1.47(m，2H).
LCMS m/z＝745.4[M+1].
Compound 74
Figure GPA0000261095610001442
1H NMR(400MHz，DMSO-d6)δ9.76(br，1H)，8.08(dd，1H)，7.19(dd，1H)，7.15-7.04(m，3H)，6.68(dd，1H)，6.60(d，1H)，6.52-6.46(m，1H)，6.42(d，1H)，4.47(s，2H)，3.70-3.64(m，2H)，3.64-3.54(m，4H)，3.50-3.46(m，2H)，3.41(s，4H)，3.21-3.16(m，1H)，3.15-3.10(m，1H)，2.76(t，2H)，2.67-2.52(m，7H)，2.46-2.40(m，3H)，2.27(t，2H)，1.72(d，3H)，1.68-1.58(m，3H)，1.58-1.52(m，2H)，1.51-1.42(d，4H)，1.26-1.22(m，1H).
LCMS m/z＝380.3[M/2+1]。
Compound 75
Figure GPA0000261095610001443
1H NMR(400MHz，DMSO-d6)δ10.05(br，1H)，8.18(d，1H)，7.89(d，1H)，7.22-7.16(m，1H)，7.13(s，1H)，7.08(dd，3H)，6.91(d，1H)，6.53(s，1H)，6.48(d，1H)，6.28(dd，1H)，5.03-4.96(m，1H)，3.69-3.64(m，2H)，3.63-3.54(m，4H)，3.41(s，4H)，3.34(s，2H)，3.32-3.18(m，5H)，2.76(t，2H)，2.72-2.57(m，4H)，2.48-2.38(m，4H)，2.27(t，2H)，1.85-1.77(m，1H)，1.73(d，2H)，1.54(t，2H)，1.27-1.19(m，1H)，0.97-0.93(m，4H)，0.76-0.71(m，2H).
LCMS m/z＝753.5[M+1].
Compound 76
Figure GPA0000261095610001451
1H NMR(400MHz，DMSO-d6)δ9.77(br，1H)，8.08(dd，1H)，7.23-7.16(m，1H)，7.10(dd，3H)，6.69(dd，1H)，6.60(d，1H)，6.51-6.46(m，1H)，6.42(dd，1H)，4.47(d，2H)，3.70-3.64(m，2H)，3.63-3.53(m，4H)，3.50-3.46(m，2H)，3.44-3.38(m，4H)，3.30(dd，3H)，2.91(s，1H)，2.81-2.74(m，3H)，2.68-2.52(m，6H)，2.45-2.39(m，2H)，2.27(t，2H)，1.72(d，2H)，1.58-1.47(m，2H)，1.28-1.20(m，2H).
LCMS m/z＝353.1[M/2+1].
Compound 77
Figure GPA0000261095610001452
1H NMR(400MHz，DMSO-d6)δ9.77(br，1H)，7.89(d，1H)，7.23-7.16(m，1H)，7.11(dd，3H)，6.60(d，1H)，6.54(s，1H)，6.42(dd，1H)，6.29(dd，1H)，4.47(d，2H)，3.71-3.63(m，2H)，3.63-3.53(m，4H)，3.45-3.38(m，4H)，3.34(s，2H)，3.32-3.26(m，5H)，2.79-2.74(m，3H)，2.67-2.60(m，3H)，2.58-2.53(m，2H)，2.45-2.40(m，2H)，2.28(t，2H)，1.84-1.78(m，1H)，1.77-1.69(m，2H)，1.54(t，2H)，1.25-1.22(m，2H)，0.97-0.93(m，2H)，0.76-0.72(m，2H).
LCMS m/z＝364.3[M/2+1].
Compound 78
Figure GPA0000261095610001453
Compound 73(0.27g, 0.36mmol) was dissolved in methanol (10mL), palladium on carbon (0.1g) was added, and the mixture was stirred at room temperature for 4 hours under a hydrogen atmosphere. The reaction mixture was filtered through celite, concentrated under reduced pressure, and separated by column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 78 as a yellow solid.
1H NMR(400MHz，CD3OD)δ8.39(d，1H)，7.62(d，1H)，7.47(dd，1H)，7.41-7.31(m，4H)，7.29(d，1H)，7.22(dd，1H)，7.02(d，1H)，6.65(d，1H)，5.38(dd，1H)，4.31(s，2H)，3.98-3.94(m，2H)，3.79-3.62(m，6H)，3.43-3.32(m，5H)，3.28-3.21(m，6H)，3.15(s，1H)，3.09(s，2H)，3.00-2.75(m，3H)，2.66(t，2H)，2.46(d，2H)，1.81(t，2H)，1.33-1.26(s，1H).
LCMS m/z＝357.7[M/2+1].
Compound 79
Figure GPA0000261095610001461
1H NMR(400MHz，CD3OD)δ8.27(d，1H)，7.43-7.30(m，6H)，7.00(s，1H)，6.86(d，1H)，5.17(dd，1H)，4.55(d，1H)，4.29(s，3H)，4.14(dd，1H)，3.86-3.81(m，3H)，3.77-3.65(m，7H)，3.63-3.52(dd，6H)，3.50(dd，1H)，3.34(s，1H)，2.90(s，3H)，2.63(dt，3H)，2.23(d，3H)，2.10-1.90(m，1H)，1.77(t，3H).
LCMS m/z＝386.2[M/2+1].
Compound 80
Figure GPA0000261095610001462
LCMS m/z＝760.4[M+1].
Compound 81
Figure GPA0000261095610001463
LCMS m/z＝368.8[M/2+1].
Compound 82
Figure GPA0000261095610001464
LCMS m/z＝360.8[M/2+1].
Compound 83
Figure GPA0000261095610001465
LCMS m/z＝703.5[M+1]。
Compound 84
Figure GPA0000261095610001471
LCMS m/z＝774.3[M+1]。
Compound 85
Figure GPA0000261095610001472
LCMS m/z＝772.4[M+1].
Compound 86
Figure GPA0000261095610001473
LCMS m/z＝367.8[M/2+1]。
Compound 87
Figure GPA0000261095610001474
LCMS m/z＝719.3[M+1].
Compound 88
Figure GPA0000261095610001475
LCMS m/z＝701.4[M+1].
Compound 89
Figure GPA0000261095610001476
LCMS m/z＝706.4[M+1].
Compound 90
Figure GPA0000261095610001481
LCMS m/z＝716.3[M+1].
Compound 91
Figure GPA0000261095610001482
1H NMR(400MHz，CD3OD)δ8.29(d，2H)，7.25-7.05(m，4H)，6.67(d，1H)，6.56(t，1H)，6.44(d，1H)，4.52(d，2H)，4.33-4.07(m，1H)，3.64-3.74(m，10H)，3.48(d，2H)，3.33(d，2H)，，2.73-2.85(m，8H)，2.56-2.65(m，2H)，2.50(s，2H)，2.43(d，2H)，1.72-1.83(m，6H)，1.66-1.43(m，6H).
LCMS m/z＝371.7[M/2+1].
Compound 92
Figure GPA0000261095610001483
1H NMR(400MHz，CD3OD)δ7.96-7.95(d，1H)，7.20-7.13(m，4H)，6.68-6.59(m，3H)，6.45-6.43(d，1H)，4.54-4.50(d，2H)，3.78-3.68(m，6H)，3.52-3.42(m，6H)，2.84(s，2H)，2.75-2.65(m，10H)，2.59-2.54(m，3H)，2.45-2.41(m，2H)，1.92-1.89(m，2H)，1.64-1.61(m，2H)，1.23-1.21(m，6H).
LCMS m/z＝365.3[M/2+1].
Compound 93
Figure GPA0000261095610001484
1H NMR(400MHz，CD3OD)δ8.39-8.35(m，1H)，8.06-8.05(d，1H)，7.53-7.51(m，1H)，7.25-7.13(m，5H)，6.98-6.96(d，1H)，6.77-6.75(d，1H)，6.64-6.61(m，2H)，5.21-5.18(m，1H)，3.76-3.64(m，7H)，3.48-3.36(m，8H)，2.86-2.76(m，6H)，2.58-2.53(m，4H)，2.48-2.40(m，2H)，1.90-1.88(m，2H)，1.54-1.40(m，5H)，1.35-1.26(m，2H)，0.94-0.88(m，3H).
LCMS m/z＝741.3[M+1].
Compound 94
Figure GPA0000261095610001491
1H NMR(400MHz，CDCl3)δ8.00-7.98(m，1H)，7.53-7.48(m，2H)，7.39-7.35(m，2H)，7.16-7.07(m，2H)，6.98-6.95(m，2H)，6.89-6.80(m，2H)，6.52-6.45(m，1H)，5.20-5.12(m，1H)，3.75-3.14(m，15H)，2.94-2.59(m，16H)，1.92-1.89(m，2H)，1.52-1.43(m，2H)，1.29-1.26(m，4H)，0.92-0.88(m，3H)
LCMS m/z＝807.5[M+1].
Compound 95
Figure GPA0000261095610001492
1H NMR(400MHz，CD3OD)δ7.74(d，1H)，7.30-7.19(m，5H)，7.12(s，1H)，6.80(d，1H)，6.57(d，1H)，6.37(d，1H)，4.33(s，2H)，4.14(s，2H)，3.83-3.80(m，2H)，3.58-3.48(m，11H)，3.42(t，3H)，3.15-3.08(m，2H)，3.02(t，2H)，2.77(t，2H)，2.68(t，2H)，2.43(t，2H)，2.35(s，3H)，2.10(d，2H)，1.98(dd，2H)，1.81(d，2H)，1.68(d，2H)，1.59-1.46(m，5H).
LCMS m/z＝378.3[M/2+1].
Compound 96
Figure GPA0000261095610001493
1H NMR(400MHz，CD3OD)δ8.39-8.35(m，1H)，7.96-7.95(d，1H)，7.22-7.16(m，5H)，6.98-6.96(d，1H)，6.65-6.58(m，3H)，5.20-5.17(m，1H)，3.78-3.64(m，6H)，3.51-3.35(m，9H)，2.86-2.77(m，6H)，2.60-2.52(m，4H)，2.48-2.42(m，2H)，1.93-1.89(m，2H)，1.68-1.62(m，2H)，1.40-1.38(m，4H)，1.23-1.21(m，6H).
LCMS m/z＝378.3[M/2+1].
Compound 97
Figure GPA0000261095610001494
1H NMR(400MHz，CD3OD)δ7.97-7.96(d，1H)，7.30-7.23(m，5H)，6.75-6.72(d，1H)，6.65-6.61(m，2H)，6.50-6.48(m，1H)，4.54-4.50(m，2H)，3.81-3.71(m，7H)，3.52-3.43(m，6H)，3.15-3.07(m，3H)，2.87-2.67(m，10H)，2.15-1.26(m，16H)，1.24-1.22(m，6H).
LCMS m/z＝783.5[M+1].
Compound 98
Figure GPA0000261095610001501
LCMS m/z＝729.5[M+1].
Compound 99
Figure GPA0000261095610001502
LCMS m/z＝359.3[M/2+1].
Compound 100
Figure GPA0000261095610001503
LCMS m/z＝372.3[M/2+1].
Compound 101
Figure GPA0000261095610001504
LCMS m/z＝392.3[M/2+1].
Compound 102
Figure GPA0000261095610001505
LCMS m/z＝386.3[M/2+1].
Compound 103
Figure GPA0000261095610001511
LCMS m/z＝357.3[M/2+1].
Compound 104
Figure GPA0000261095610001512
LCMS m/z＝753.5[M+1].
Compound 105
Figure GPA0000261095610001513
LCMS m/z＝371.3[M/2+1].
Compound 106
Figure GPA0000261095610001514
LCMS m/z＝755.3[M+1]。
Compound 107
Figure GPA0000261095610001515
LCMS m/z＝372.2[M/2+1].
Compound 108
Figure GPA0000261095610001516
LCMS m/z＝405.3[M/2+1]。
Compound 109
Figure GPA0000261095610001521
LCMS m/z＝399.3[M/2+1]。
Compound 110
Figure GPA0000261095610001522
LCMS m/z＝352.7[M/2+1]。
Compound 111
Figure GPA0000261095610001523
LCMS m/z＝385.8[M/2+1]。
Compound 112
Figure GPA0000261095610001524
1H NMR(400MHz，CDCl3)δ7.46(d，1H)，7.21-7.04(m，4H)，6.69(d，1H)，6.54(s，1H)，6.24(d，1H)，4.54(d，2H)，4.09(s，1H)，3.98-3.88(m，3H)，3.81-3.52(m，9H)，3.47(s，3H)，3.38(d，1H)，3.31-3.23(m，1H)，3.22-3.17(m，1H)，2.91-2.65(m，8H)，2.62-2.46(m，5H)，2.38(s，1H)，1.83(s，2H)，1.71(s，1H)，1.61-1.46(m，3H)，1.32-1.18(m，3H)，0.63(s，2H)，0.35(s，2H).
LCMS m/z＝393.8[M/2+1]
Compound 113
Figure GPA0000261095610001525
1H NMR(400MHz，CD3OD)δ8.43(d，1H)，7.93(d，1H)，7.43-7.35(m，4H)，7.33(d，1H)，7.17(s，1H)，7.07(d，1H)，7.00(d，1H)，6.69(d，1H)，5.44(dd，1H)，4.32(d，3H)，3.99-3.94(m，2H)，3.79(t，2H)，3.71(dt，5H)，3.61(d，3H)，3.38(s，2H)，3.27(d，4H)，3.04(dt，1H)，2.93(t，2H)，2.77(t，2H)，2.30-2.22(m，2H)，2.06(d，1H)，1.93(s，4H)，1.79(s，2H)，1.71-1.50(m，5H)，1.38-1.25(m，7H).
LCMS m/z＝398.3[M/2+1].
Compound 114
Figure GPA0000261095610001531
LCMS m/z＝702.5[M+1]。
Compound 115
Figure GPA0000261095610001532
1H NMR(400MHz，DMSO-d6)δ10.14(br，1H)，8.17(d，1H)，7.94(d，1H)，7.20(dd，2H)，7.08-7.02(m，2H)，6.91(d，1H)，6.65(s，1H)，6.48(dd，2H)，5.02-4.97(m，1H)，3.69-3.64(m，2H)，3.62-3.54(m，4H)，3.49(s，2H)，3.43-3.39(m，2H)，3.38-3.10(m，12H)，2.81(t，2H)，2.72-2.61(m，4H)，2.32(t，2H)，1.74(d，2H)，1.53(t，2H)，1.23(d，3H).
LCMS m/z＝366.3[M/2+1].
Compound 116
Figure GPA0000261095610001533
1H NMR(400MHz，DMSO-d6)δ8.17(dd，1H)，7.81(d，1H)，7.20(d，2H)，7.05(t，2H)，6.91(d，1H)，6.56(s，1H)，6.48(dd，1H)，5.00(t，1H)，4.01(d，2H)，3.91(s，2H)，3.70-3.52(m，8H)，3.48(d，2H)，3.43(s，1H)，3.30(t，2H)，2.91(s，1H)，2.80(s，2H)，2.77(s，1H)，2.72-2.63(m，4H)，2.53(d，1H)，2.47(s，1H)，2.32(s，4H)，1.66(s，2H)，1.50(s，2H)，1.23(d，2H)，0.53(d，2H)，0.37(s，2H).
LCMS m/z＝394.8[M/2+1].
Compound 117
Figure GPA0000261095610001534
1H NMR(400MHz，CD3OD)δ8.39-8.34(m，1H)，8.26-8.25(m，1H)，7.19-7.13(m，5H)，7.00-6.98(m，2H)，6.81-6.80(d，1H)，6.65-6.61(m，1H)，5.21-5.16(m，1H)，3.71-3.64(m，6H)，3.55-3.46(m，8H)，3.02(s，2H)，2.84-2.75(m，7H)，2.63-2.60(m，4H)，2.46-2.41(m，2H)，1.90-1.86(m，2H)，1.63-1.61(m，2H).
LCMS m/z＝384.3[M/2+1].
Compound 118
Figure GPA0000261095610001541
1H NMR(400MHz，CD3OD)δ7.72(s，1H)，7.23(dd，2H)，7.08(t，1H)，6.74(d，1H)，6.62(d，1H)，6.50(d，1H)，4.62(s，1H)，4.30-4.20(m，1H)，4.03(d，2H)，3.98(d，2H)，3.80-3.57(m，11H)，3.51(t，2H)，3.12(dt，3H)，2.95-2.85(m，4H)，2.70-2.64(m，3H)，2.56(d，3H)，1.88(s，4H)，1.81-1.55(m，8H)，1.51(dd，2H)，0.61(d，2H)，0.40(d，2H).
LCMS m/z＝415.8[M/2+1]。
Compound 119
Figure GPA0000261095610001542
1H NMR(400MHz，DMSO-d6)δ10.21(br，1H)，8.29(d，1H)，8.18(dd，1H)，7.18(dd，1H)，7.13(s，1H)，7.11-7.05(m，3H)，6.92(dd，1H)，6.83(d，1H)，6.49(dd，1H)，5.15(s，1H)，5.03(dd，1H)，3.70(d，2H)，3.67-3.53(m，9H)，3.41(d，2H)，3.38-3.29(m，3H)，2.92(s，1H)，2.79-2.66(m，8H)，2.64(d，1H)，2.35(s，4H)，1.67(d，2H)，1.54(d，2H)，1.36(s，6H).
LCMS m/z＝379.8[M/2+1].
Compound 120
Figure GPA0000261095610001543
1H NMR(400MHz，DMSO-d6)δ10.26(br，1H)，8.42(d，1H)，8.17(dd，1H)，8.13(dd，2H)，7.54-7.49(m，3H)，7.21-7.16(m，2H)，7.13-7.05(m，4H)，6.92(dd，1H)，6.48(dd，1H)，5.02(s，1H)，3.81(d，2H)，3.76-3.67(m，4H)，3.57(dt，5H)，3.42(d，3H)，3.30(t，6H)，2.91(s，1H)，2.80-2.65(m，5H)，2.42-2.29(m，4H)，1.72(d，2H)，1.57(t，2H).
LCMS m/z＝388.8[M/2+1]。
Compound 121
Figure GPA0000261095610001551
LCMS m/z＝782.3[M+1]。
Compound 122
Figure GPA0000261095610001552
LCMS m/z＝394.8[M/2+1]。
Compound 123
Figure GPA0000261095610001553
LCMS m/z＝729.4[M+1]。
Compound 124
Figure GPA0000261095610001554
1H NMR(400MHz，DMSO-d6)δ9.81(br，1H)，7.97(d，1H)，7.19(d，1H)，7.16-7.06(m，3H)，6.87(d，1H)，6.66(s，1H)，6.51(dd，2H)，4.83(d，1H)，4.47(s，2H)，3.68(s，2H)，3.58(dd，4H)，3.42(s，4H)，3.37-3.17(m，12H)，2.77(t，3H)，2.65(s，3H)，2.56(d，3H)，2.43(d，2H)，2.28(t，2H)，1.75(d，3H)，1.68-1.43(m，9H).
LCMS m/z＝400.3[M/2+1]。
Compound 125
Figure GPA0000261095610001555
1H NMR(400MHz，CD3OD)δ8.38-8.34(m，1H)，7.88-7.86(d，1H)，7.22-7.14(m，5H)，6.98-6.96(d，1H)，6.65-6.62(m，1H)，6.52(s，1H)，6.33-6.31(m，1H)，5.20-5.16(m，1H)，3.77-3.65(m，6H)，3.53-3.40(m，8H)，3.02(s，2H)，2.84-2.75(m，7H)，2.58-2.55(m，4H)，2.46-2.41(m，2H)，1.88-1.84(m，2H)，1.67-1.64(m，2H)，1.04-1.02(m，2H)，0.77-0.74(m，2H).
LCMS m/z＝370.3[M/2+1]
Compound 126
Figure GPA0000261095610001561
1H NMR(400MHz，CD3OD)δ8.38-8.34(m，1H)，7.89-7.88(d，1H)，7.22-7.11(m，5H)，6.98-6.96(d，1H)，6.81-6.75(m，2H)，6.63-6.61(m，1H)，5.21-5.16(m，1H)，3.71-3.64(m，7H)，3.52-3.26(m，6H)，3.02(s，2H)，2.84-2.75(m，8H)，2.58-2.55(m，4H)，2.39-2.36(m，2H)，1.88-1.84(m，2H)，1.64-1.56(m，2H).
LCMS m/z＝733.3[M+1].
Compound 127
Figure GPA0000261095610001562
1H NMR(400MHz，DMSO-d6)δ9.83(br，1H)，7.96(d，1H)，7.21-7.17(m，1H)，7.15-7.07(m，3H)，6.87(d，1H)，6.66(s，1H)，6.54-6.46(m，2H)，4.86(s，1H)，4.52-4.44(m，2H)，3.67(d，2H)，3.59(dd，5H)，3.43(s，5H)，3.36-3.17(m，8H)，2.77(t，2H)，2.68(d，4H)，2.58(t，2H)，2.43(d，2H)，2.29(t，2H)，1.75(d，3H)，1.65(s，3H)，1.53(dd，7H).
LCMS m/z＝393.4[M/2+1]。
Compound 128
Figure GPA0000261095610001563
1H NMR(400MHz，CD3OD)δ8.26-8.25(d，1H)，7.23-7.13(m，4H)，6.96-6.91(m，2H)，6.81-6.80(d，1H)，6.72-6.70(m，1H)，4.76-4.72(m，1H)，3.78-3.66(m，6H)，3.57-3.53(m，8H)，3.01(s，2H)，2.84-2.75(m，7H)，2.63-2.57(m，4H)，2.45-2.40(m，2H)，1.90-1.86(m，2H)，1.63-1.61(m，2H).
LCMS m/z＝387.3[M/2+1].
Compound 129
Figure GPA0000261095610001571
1H NMR(400MHz，CD3OD)δ8.39-8.34(m，1H)，8.04-8.02(m，1H)，7.21-7.13(m，5H)，6.98-6.93(d，1H)，6.64-6.62(m，1H)，6.54-6.40(m，2H)，5.21-5.16(m，1H)，3.75-3.66(m，6H)，3.53-3.44(m，8H)，3.02(s，2H)，2.84-2.75(m，7H)，2.63-2.60(m，4H)，2.46-2.41(m，2H)，1.89-1.85(m，2H)，1.65-1.60(m，2H).
LCMS m/z＝718.4[M+1].
Compound 130
Figure GPA0000261095610001572
LCMS m/z＝349.8[M/2+1]。
Compound 131
Figure GPA0000261095610001573
LCMS m/z＝350.3[M/2+1]。
Compound 132
Figure GPA0000261095610001574
LCMS m/z＝396.8[M/2+1]。
Compound 133
Figure GPA0000261095610001575
LCMS m/z＝357.8[M/2+1]。
Compound 134
Figure GPA0000261095610001581
LCMS m/z＝379.8[M+1]。
Compound 135
Figure GPA0000261095610001582
LCMS m/z＝396.2[M+1]。
Compound 136
Figure GPA0000261095610001583
1H NMR(400MHz，CD3OD)δ8.35(dd，1H)，8.28(d，2H)，7.25-7.05(m，5H)，6.96(d，1H)，6.62(dd，1H)，6.55(t，1H)，5.16-5.21(m，1H)，3.77-3.62(m，10H)，3.56-3.41(m，4H)，3.01(s，2H)，2.92-2.73(m，7H)，2.67-2.48(m，4H)，2.43(t，2H)，1.81(d，2H)，1.70-1.53(m，2H).
LCMS m/z＝351.0[M/2+1].
Compound 137: 7- [ (1R) -2- [2- [ 3-fluoro-5- [ [4- (4-methyl-2-pyridinyl) -1-oxo-4, 9-diazaspiro [5.5] deca-alk-9-yl ] methyl ] phenyl ] ethylamino ] -1-hydroxy-ethyl ] -4-hydroxy-3H-1, 3-benzothiazol-2-one
7-[(1R)-2-[2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethylamino]-1-hydroxy-ethyl]-4-hydroxy-3H-1，3-benzothiazol-2-one
Figure GPA0000261095610001584
The first step is as follows: 2- [ 3-fluoro-5- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethanol (137C)
2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethanol
137A (0.61g, 2.5mmol) was dissolved in acetonitrile (30mL), and 137B (0.63g, 2.7mmol), potassium carbonate (0.68g, 4.9mmol) and water (1mL) were added to react at 50 ℃ for 2.5 hours. Water (100mL) was added, extraction was performed with DCM (100mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 15: 1) to give 137C as a yellow liquid (0.80g, yield 81%).
The second step is that: 9- [ [3- (2-bromoethyl) -5-fluoro-phenyl ] methyl ] -4- (4-methyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecane (137D)
9-[[3-(2-bromoethyl)-5-fluoro-phenyl]methyl]-4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecane
137C (0.80g, 2.0mmol) was dissolved in DCM (30mL), and imidazole (0.2g, 3.0mmol), carbon tetrabromide (0.80g, 2.4mmol), triphenylphosphine (0.79g, 3.0mmol) and reacted at room temperature for 5 hours. Water (100mL) was added to the reaction solution, followed by extraction, and aqueous phase was extracted with DCM (50mL × 2), and the combined organic phases were washed with saturated brine (100mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM/methanol (v/v) ═ 20: 1) to give 137D (0.45g, yield 49%) as a yellow liquid.
The third step: 7- [ (1R) -1- [ tert-butyl (dimethyl) silyl ] oxy-2- [2- [ 3-fluoro-5- [ [4- (4-methyl-2-pyridyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethylamino ] ethyl ] -4-hydroxy-3H-1, 3-benzothiazol-2-one (137E)
7-[(1R)-1-[tert-butyl(dimethyl)silyl]oxy-2-[2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethylamino]ethyl]-4-hydroxy-3H-1，3-benzothiazol-2-one
137D (0.45g, 0.97mmol) was dissolved in DMF (30mL), 7- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -4-hydroxy-3H-1, 3-benzothiazol-2-one (0.30g, 0.88mmol), diisopropylethylamine (0.72g, 5.6mmol) was added, reacted at 50 ℃ for 40 hours, the reaction was directly concentrated under reduced pressure and column chromatography (DCM/methanol (v/v): 10: 1) gave 137E (0.12g, 19% yield) as a yellow solid.
The fourth step: 7- [ (1R) -2- [2- [ 3-fluoro-5- [ [4- (4-methyl-2-pyridinyl) -1-oxa-4, 9-diazaspiro [5.5] undecan-9-yl ] methyl ] phenyl ] ethylamino ] -1-hydroxy-ethyl ] -4-hydroxy-3H-1, 3-benzothiazol-2-one (compound 137)
7-[(1R)-2-[2-[3-fluoro-5-[[4-(4-methyl-2-pyridyl)-1-oxa-4，9-diazaspiro[5.5]undecan-9-yl]methyl]phenyl]ethylamino]-1-hydroxy-ethyl]-4-hydroxy-3H-1，3-benzothiazol-2-one
137E (0.12g, 0.17mmol) was dissolved in THF (10mL), triethylamine trihydrofluoric acid (0.5g, 3.0mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction was quenched by the addition of saturated aqueous sodium bicarbonate (50mL) and methanol (5mL) in that order, the residue was extracted with DCM (100mL × 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure and separated by column chromatography (DCM: methanol (v/v) ═ 1: 0 to 8: 1) to give compound 137(0.03g, yield 30%) as a yellow solid.
LCMS m/z＝304.7[M/2+1].
Compound 138
Figure GPA0000261095610001591
Compound 139
Figure GPA0000261095610001601
1H NMR(400MHz，DMSO-d6)δ8.18(d，1H)，7.22-7.17(td，1H)，7.14(s，1H)，7.08(dd，3H)，6.91(d，1H)，6.88-6.74(m，4H)，6.48(d，1H)，5.05-4.96(m，1H)，3.75-3.69(m，2H)，3.67(s，3H)，3.63-3.54(m，4H)，3.42(d，2H)，3.31(dd，2H)，2.93-2.88(m，3H)，2.80(s，2H)，2.78-2.75(m，3H)，2.73-2.62(m，4H)，2.54(d，1H)，2.46(dd，3H)，2.28(t，2H)，1.84(d，2H)，1.58(t，2H)，1.23(s，2H).
LCMS m/z＝364.8[M/2+1].
Compound 140
Figure GPA0000261095610001602
LCMS m/z＝351.8[M/2+1]。
Compound 141
Figure GPA0000261095610001603
1H NMR(400MHz，DMSO-d6)δ8.40(dd，1H)，8.17(d，1H)，8.05(dd，1H)，7.23-7.16(m，1H)，7.14-7.04(m，4H)，6.94-6.89(m，2H)，6.48(dd，1H)，5.00(dd，1H)，3.77-3.72(m，2H)，3.63-3.50(m，7H)，3.47(s，2H)，3.41(d，2H)，3.38-3.25(m，4H)，2.80-2.72(m，4H)，2.71-2.57(m，4H)，2.54(d，1H)，2.47-2.42(m，2H)，2.26(t，2H)，1.84-1.76(m，2H)，1.55(t，2H).
LCMS m/z＝362.8[M/2+1]。
Compound 142
Figure GPA0000261095610001604
1H NMR(400MHz，DMSO-d6)δ10.22(s，1H)，8.17(d，1H)，7.39(dd，1H)，7.31-7.26(m，1H)，7.22-7.16(m，1H)，7.16-7.00(m，6H)，6.91(d，1H)，6.52-6.43(m，1H)，5.00(dd，1H)，3.79-3.72(m，2H)，3.62-3.54(m，4H)，3.41(d，2H)，3.35-3.25(m，4H)，2.92-2.88(m，3H)，2.76(d，5H)，2.73-2.60(m，4H)，2.54(d，1H)，2.48-2.40(m，3H)，2.30(t，2H)，1.92(d，2H)，1.61(t，2H).
LCMS m/z＝366.8[M/2+1]。
Compound 143
Figure GPA0000261095610001611
LCMS m/z＝741.4[M+1].
Compound 144
Figure GPA0000261095610001612
LCMS m/z＝335.1[M/2+1].
Compound 145
Figure GPA0000261095610001613
LCMS m/z＝834.4[M+1].
Compound 146
Figure GPA0000261095610001614
LCMS m/z＝744.4[M+1].
Compound 145(0.41g, 0.5mmol) was dissolved in methanol (10mL), 10% palladium on carbon (100mg) was added, and the mixture was stirred under a hydrogen atmosphere at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure and subjected to column chromatography (dichloromethane/methanol (v/v) ═ 4: 1) to give compound 146(0.37g, yield 50%) as a yellow solid.
Compound 147
Figure GPA0000261095610001615
LCMS m/z＝303.7[M/2]
Adopting a synthesis method (same solvent, temperature, reagent and feeding ratio) similar to that of the compound 137, 137B is used
Figure GPA0000261095610001621
Instead.
Compound 148
Figure GPA0000261095610001622
Adopting a synthesis method (same solvent, temperature, reagent and feeding ratio) similar to that of the compound 137, 137B is used
Figure GPA0000261095610001623
Instead, 7- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl]Oxoethyl radical]Use of (E) -4-hydroxy-3H-1, 3-benzothiazol-2-one
Figure GPA0000261095610001624
Instead.
Biological test example
Test example 1: inhibitory Activity on human muscarinic M3 receptor
CHO cells (PerkinElmer, ES-212-AF) stably expressing human muscarinic receptor 3(hM3) and apo-Aequorin were cultured in Ham' S F12 medium (Invitrogen 12500 062) containing 10% Fetal Bovine Serum (FBS) (Gibico 10099-141), 400. mu.g/mL G418(sigmaG5013) and 250. mu.g/mLzeocin (Invitrogen ant-zn-5p) at 37 ℃ with 5% CO2Culturing under the condition to achieve 90-100% fusion. The cells were separated by PBS/5mM EDTA rinse, harvested by centrifugation, resuspended and counted in phenol-free Ham's F12 medium (Invitrogen 11039-6cells/mL. 15mL of the cell suspension was added to a 50mL centrifuge tube and Coelenterazine-h (Promega S2011) was added to a final concentration of 5. mu.M. Wrapped with tinfoil and protected from light, and incubated for 4 hours at 20 ℃ on a rotary shaker. The cells were then diluted with 0.1% BSA/phenol red free Ham's F12 medium to a final concentration of 5.0X 10 5cells/mL, cells were placed on a rotary shaker and spun at low speed and incubated at room temperature for at least 1 hour. The compounds of the examples were formulated in DMSO as 10mM stock solution, 0.1% BSA/phenol red free Ham's F12 medium diluted in gradient (log (M): 7, -8, -9, -10, -11), and added to 96 well plates at 50. mu.L per well. A further 50. mu.L of cell suspension (25000 cells/well) was added to each well and incubated for 15 minutes at room temperature. The 96-well plate was placed in a microplate reader (Perkin Elmer, Envision), 50. mu.L of acetylcholine chloride (SigmaA6625) solution at a concentration of 112.92nM (hM3) was added to each well with a microplate reader applicator, luminescence was recorded for 20 seconds, and IC was calculated and analyzed using origin7.550. Inhibitory Activity of the Compounds of the invention at human muscarinic receptors the IC determined by the above assay50The values are given in table 1 below. TABLE 1 testingResults of the inhibitory Activity of Compounds on human muscarinic M3 receptors
Figure GPA0000261095610001625
Figure GPA0000261095610001631
And (4) conclusion: the invention has obvious inhibitory activity on human muscarinic M3 receptor.
Test example 2: agonistic activity at the human adrenergic beta 2 receptor
Agonist activity of the compounds of the examples on human adrenergic receptors was determined by LANCE Ultra cAMP Assay.
CHO cells (PerkinElmer, ES-034-CF) stably expressing human adrenergic receptor (h.beta.2) were cultured in MEM-alpha medium (Invitrogen 12561-056) containing 10% Fetal Bovine Serum (FBS) (Gibico 10099-141) and 250. mu.g/mL Zeocin (Invivogen ant-zn-5p) at 37 ℃ with 5% CO 2Culturing under the condition, and detecting the cAMP agonism of the examples by using a LANCE Ultra cAMP Assay kit (PerkinElmer TRF0263) after 90-100% fusion is achieved. The cells were separated with PBS/5mM EDTA, collected by centrifugation, resuspended in Stimulation Buffer (1XHBSS, 5mM HEPES, 0.5mM IBMX, 0.1% BSA, pH7.4), and adjusted to 6X 10 cell concentration5cells/ml. The compounds of the examples were made up in DMSO as 10mM stock, diluted in a Ststimulation Buffer gradient and added to 384 well plates at 5. mu.l per well. mu.L of cell suspension (3000 cells/well) was added to each well, and after incubation for 30 minutes at room temperature, 5. mu.l of 4 × Eu-cAMP tracer working solution was added to each well, followed by 5. mu.l of 4 × Ulight-anti-cAMP working solution to each well, and incubation for 1 hour at room temperature was carried out. The TR-FRET was detected in 384 well plates using a microplate reader (Perkin Elmer, Envision) and EC was calculated and analyzed using origin7.550. The agonistic activity of the compound of the present invention on human adrenergic receptors was determined by the above experiment, and the EC was measured50The values are shown in Table 2:
TABLE 2 results of agonist activity of test compounds at the human adrenergic beta 2 receptor
Figure GPA0000261095610001641
Figure GPA0000261095610001651
And (4) conclusion: the invention has obvious agonist activity on beta 2 adrenergic receptors.
Test example 3: inhibition of methacholine-induced bronchoconstriction in guinea pigs
The 8-week-old male guinea pigs were purchased in the Witonglihua and the experiment was started after 3 days of acclimation. The stock solution to be tested was prepared with 83% absolute ethanol and 17% tween 80 to 0.6 mM. Diluted with water to the desired concentration prior to administration. Before administration, animals were anesthetized with 5% isoflurane using a small animal anesthesia machine (Matrx; VME2) for 1.5-2 minutes. After anesthetizing the guinea pigs, the guinea pigs were mounted on a tracheal intubation operating platform and administered intratracheally using a rat liquid aerosol dosing kit (pen-centre; MSA-250-R) in a volume of 250. mu.l per guinea pig. After administration, guinea pig enhanced expiratory pause (enhanced pause; PenH) values were measured at 4 hours, 24 hours using a full volume profiler (DSI; GS220A 12-R7B). Nebulization was given at 3mg/ml methacholine (Mch) for 36 seconds and recording time 7 minutes. The PenH mean was calculated. (reference J Pharmacol Exp Ther 345: 260-270.). The results are shown in Table 3.
PenH formula: PenH PEP/PIP Pause; pause ═ Te-Tr/Tr
Te: expiratory phase time(s)
Tr: relaxation phase time(s)
PEP: expiratory peak flow rate (ml/s)
PIP: inspiratory peak flow rate (ml/s)
TABLE 3 inhibition of acetylcholine-induced bronchoconstriction in guinea pigs
Figure GPA0000261095610001652
Figure GPA0000261095610001661
"-" represents no test.
And (4) conclusion: the compound of the invention has obvious inhibition effect on acetylcholine-induced bronchoconstriction of guinea pigs, and after being administrated for 24 hours, part of the compound still has good bronchoconstriction inhibition effect.

Claims (8)

1. A compound of formula (II) or formula (III), a stereoisomer or a pharmaceutically acceptable salt thereof:
Figure FDA0003358962110000011
R1selected from the group consisting of a benzene ring, a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, an isothiazole ring, an isoxazole ring, a pyrazole ring, a pyridine ring, a pyrimidine ring, a pyrazine ring, a quinoline ring, an isoquinoline ring, a benzisothiazole ring, an indazole ring, a 1H-pyrazolo [4,3-b ] ring]A pyridine ring or
Figure FDA0003358962110000012
The benzene ring, the thiophene ring, the furan ring, the pyrrole ring, the thiazole ring, the oxazole ring, the imidazole ring, the isothiazole ring, the isoxazole ring, the pyrazole ring, the pyridine ring, the pyrimidine ring, the pyrazine ring, the quinoline ring, the isoquinoline ring, the benzisothiazole ring, the indazole ring, the 1H-pyrazole [4,3-b ]]A pyridine ring or
Figure FDA0003358962110000013
Optionally further substituted by 0 to 5R1aSubstitution;
R1aselected from F, Cl, Br, nitro, amino, cyano, hydroxy, carboxy, -C (═ O) OBn, -CF3Methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, ethynyl, propynyl, propargyl, and the like, Methoxy, ethoxy, propoxy, isopropoxy, hydroxymethyl, hydroxyethyl, -C (CH)3)2OH, phenoxy, anilino, phenyl, 2-thienyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or-CH2-a cyclopropyl group;
w is selected from a bond or-C (═ O) -;
provided that when W is selected from-C (═ O) -, R1By 1 to 5R1aSubstituted and has at least 1R1aSelected from ethynyl, propynyl, propargyl or-CH2-a cyclopropyl group;
Rxeach independently selected from H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl or-CH2-a cyclopentyl group;
R8selected from H or OH;
b is selected from
Figure FDA0003358962110000014
p1 is selected from 0 or 1;
p2 is selected from 0, 1, 2, 3 or 4;
Raselected from H, F, Cl, Br, I, C1-4Alkyl radical, C1-4Alkoxy or C3-6A cycloalkyl group.
2. A compound, stereoisomer, or pharmaceutically acceptable salt thereof, according to claim 1, wherein:
Raselected from F, Cl, Br, I, methyl, ethyl, methoxy, ethoxy or cyclopropyl;
b is selected from
Figure FDA0003358962110000021
3. A compound, stereoisomer or pharmaceutically acceptable salt thereof, according to claim 2, wherein:
R1-w is selected from
Figure FDA0003358962110000022
Figure FDA0003358962110000023
Figure FDA0003358962110000024
4. A compound, stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from:
Figure FDA0003358962110000031
Figure FDA0003358962110000041
Figure FDA0003358962110000051
Figure FDA0003358962110000061
Figure FDA0003358962110000071
Figure FDA0003358962110000081
Figure FDA0003358962110000091
Figure FDA0003358962110000101
Figure FDA0003358962110000111
5. A pharmaceutical composition comprising a therapeutically effective dose of a compound according to any one of claims 1 to 4, a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.
6. The pharmaceutical composition of claim 5, wherein the additional therapeutic agent is selected from one or more of a PDE4 inhibitor, an M receptor antagonist, a corticosteroid, and a beta-adrenoceptor agonist.
7. Use of a compound according to any one of claims 1 to 4, a stereoisomer or pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of claims 5 to 6, in the manufacture of a medicament for the treatment of an obstructive airways disease.
8. Use according to claim 7, wherein the obstructive airways disease is selected from the group consisting of asthma, chronic obstructive pulmonary disease and bronchitis.
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WO2012085583A1 (en) * 2010-12-23 2012-06-28 Astrazeneca Ab New compound
WO2012085582A1 (en) * 2010-12-23 2012-06-28 Astrazeneca Ab Compound
CN102625808A (en) * 2009-07-31 2012-08-01 阿斯利康(瑞典)有限公司 Spirocyclic amide derivatives
CN103249418A (en) * 2010-10-07 2013-08-14 阿斯利康(瑞典)有限公司 Novel combinations

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CN102083839A (en) * 2008-02-06 2011-06-01 阿斯利康(瑞典)有限公司 Compounds
WO2011012897A1 (en) * 2009-07-31 2011-02-03 Astrazeneca Ab New combinations for the treatment of asthma
CN102625808A (en) * 2009-07-31 2012-08-01 阿斯利康(瑞典)有限公司 Spirocyclic amide derivatives
CN103249418A (en) * 2010-10-07 2013-08-14 阿斯利康(瑞典)有限公司 Novel combinations
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