CN107108562B - With β2ReceptorsNitrogen-containing heterocyclic ring derivative with agonistic and M receptor antagonistic activities and application thereof in medicine - Google Patents

With β2ReceptorsNitrogen-containing heterocyclic ring derivative with agonistic and M receptor antagonistic activities and application thereof in medicine Download PDF

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CN107108562B
CN107108562B CN201680003435.9A CN201680003435A CN107108562B CN 107108562 B CN107108562 B CN 107108562B CN 201680003435 A CN201680003435 A CN 201680003435A CN 107108562 B CN107108562 B CN 107108562B
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hydroxy
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ethyl
azaspiro
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CN107108562A (en
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魏用刚
邱关鹏
雷柏林
楚红柱
郑苏欣
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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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/438—The ring being spiro-condensed with carbocyclic or heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4709—Non-condensed quinolines and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/536—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 ortho- or peri-condensed with carbocyclic ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Abstract

A compound shown as a general formula (I) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, a preparation method thereof and application thereof in preparing a medicament for treating airway obstruction diseases, wherein the compound shown as the general formula (I) is shown as the formula (I). Wherein the definition of each substituent is the same as that in the specification.

Description

Nitrogen-containing heterocyclic spiro derivative with β 2 receptor agonistic and M receptor antagonistic activities and application thereof in medicine
Technical Field
The invention relates to a nitrogenous heterocyclic spiro derivative, a preparation method and application thereof in medicine, in particular to a compound with muscarinic receptor antagonism and β2-dual active azaspiro derivatives of adrenergic receptor agonism or stereoisomers, hydrates, solvates, metabolites, pharmaceutically acceptable salts, co-crystals or prodrugs thereof, pharmaceutical compositions thereof and their use in medicine.
Background
Bronchodilators, which are widely used in clinics, include muscarinic receptor antagonists and β2Current inhaled muscarinic receptor antagonists include ipratropium bromide, oxitropium bromide, glycopyrrolate, tiotropium bromide, aclidinium bromide and umeclidinium bromide β2Adrenergic agonists dilate bronchi by stimulating adrenergic receptors of airway smooth muscle, reversing bronchoconstrictor pairsReaction of a mediator such as acetylcholine β 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 studies have found, the combined use of muscarinic receptor antagonist and β was demonstrated2Adrenergic 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 anoorlipta (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 composition having both muscarinic receptor antagonism and β2These compounds are administered as a single therapeutic agent, providing bronchodilatory action from two distinct and possibly synergistic modes of action2Adrenergic 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 GPA0000234146600000031
wherein:
R1is selected from
Figure GPA0000234146600000032
R1aSelected from H, C2-6Alkenyl radical, C2-6Alkynyl, C3-10Carbocyclyl, 3-to 8-membered heterocyclyl, C3-10carbocyclyl-C1-4Alkylene or 3-to 8-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1, 2 or 3 heteroatoms selected from N, O or S;
R1bis selected from C2-6Alkenyl radical, C2-6Alkynyl, C3-10Carbocyclyl, 3-to 8-membered heterocyclyl, C3-10carbocyclyl-C1-4Alkylene or 3-to 8-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1, 2 or 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy, cyano, NH2、C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio or-C (═ O) NH2NH of said2、-C(＝O)NH2Or alkyl is optionally further substituted by 0, 1 or 2 substituents selected from F, Cl, Br, I, hydroxy, cyano or C1-4Alkyl is substituted by a substituent;
m is selected from the group consisting of a bond, -CH2-、-CH2CH2-、-O-、-S-、-OCH2-、-SCH2-or-CH ═ CH-;
R1dselected from H, hydroxy, -CH2OH or C1-4An alkyl group;
R1eand R1fEach independently selected from F, Cl, Br, I, CF3、NH2OH, carboxyl, cyano, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylthio, -NHC1-4Alkyl, -N (C)1-4Alkyl radical)2、-S(＝O)-C1-4Alkyl, -S (═ O)2-C1-4Alkyl or-C (═ O) O-C1-4An alkyl group;
ring A and ring C are each independently selected from C6-10A carbocycle or a 5-to 10-membered heterocycle, said carbocycle or heterocycle optionally further substituted with 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, C1-4Alkyl or C1-4Alkoxy, and said heterocycle contains 1, 2, or 3 heteroatoms selected from N, O or S;
R2selected from the group consisting of a bond, C1-6Alkylene radical, C2-6Alkenylene or C2-6Alkynylene, said alkylene, alkenylene or alkynylene being optionally further substituted by 0, 1, 2, 3, 4 or 5 groups selected from F, Cl, Br, I, OH, cyano, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4Substituted by a substituent of alkylene;
l is selected from a bond or
Figure GPA0000234146600000041
And Y is directly connected to R2Connecting;
R3selected from the group consisting of a bond and C1-6Alkylene group ofAlkylene is optionally further selected from R by 0, 1, 2, 3, 4 or 53aSubstituted with the substituent(s);
and R is2L and R3The three can not be simultaneously keys;
R4、R5each independently selected from H or C1-4An alkyl group;
Figure GPA0000234146600000042
represents β -adrenoceptor binding group;
R3aselected from F, Cl, Br, I, cyano, OH, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4An alkylene group;
alternatively, two R3aMay form, together with the atoms to which they are attached, a 3 to 6 membered carbocyclic ring optionally further substituted by 0, 1, 2, 3, 4 or 5 substituents selected from F, Cl, Br, I, cyano, OH, C1-4Alkyl or C1-4Substituted by a substituent of alkoxy;
A1and A4Each independently selected from C3-7Cycloalkylene radical, C6-10Arylene, 5-to 8-membered heteroarylene, -O-C6-10Arylene, -O-5 to 8 membered heteroarylene, 5 to 8 membered heteroarylene-O-or C6-10arylene-O-wherein said cycloalkylene, arylene or heteroarylene is optionally further substituted with 0, 1, 2, 3, 4 or 5 substituents selected from R6Substituted with the substituent(s);
A2and A3Each independently selected from C1-6Alkylene radical, C2-6Alkenylene or C2-6Alkynylene, said alkylene, alkenylene or alkynylene being optionally further substituted by 0, 1, 2, 3, 4 or 5 groups selected from F, Cl, Br, I, OH, cyano, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4Substituted by a substituent of alkylene;
x and Y are each independently selected from the group consisting of a bond, -O-, -C (═ O) O-, -OC (═ O) -, -S (═ O)2-、-C(＝O)NR7-、-NR7C(＝O)-、-OC(＝O)NR7-、-NR7C(＝O)O-、-NR7C(＝O)NR7、-NR7S(＝O)2-、-S(＝O)2NR7-、-NR7S(＝O)2NR7or-NR7-；
R6Selected from F, Cl, Br, I, OH, NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C2-4Alkenyl radical, C2-4Alkynyl, C1-4Alkoxy, -OC3-6Cycloalkyl radical, C1-4Alkylthio, -S (═ O) -C1-4Alkyl, -S (═ O)2-C1-4Alkyl, -C (═ O) -C1-4Alkyl, -C (═ O) O-C1-4Alkyl, -OC (═ O) -C1-4Alkyl, 5-to 6-membered heteroaryl or-C (═ O) NH2The alkyl, alkoxy, cycloalkyl, alkenyl, alkynyl, heteroaryl, NH2or-C (═ O) NH2Optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CF3、C1-4Alkyl radical, C1-4Alkoxy or-C (═ O) -C1-4Alkyl is substituted by a substituent;
R7each independently selected from H or C1-4An alkyl group;
a is 0, 1, 2, 3 or 4;
b is 0, 1, 2, 3, 4 or 5;
m, n, p or q are each independently selected from 0 or 1.
In a specific embodiment of the present invention, in the compound represented by the above general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,
Figure GPA0000234146600000051
represents β -adrenoceptor binding group;
b is preferably selected from
Figure GPA0000234146600000052
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) OC1-4Alkyl, -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 selected from
Figure GPA0000234146600000053
Figure GPA0000234146600000054
Wherein Q is selected from-CH ═ CH-, -CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-；
B is further preferably selected from
Figure GPA0000234146600000055
Figure GPA0000234146600000061
In a preferred embodiment of the present invention, in the compound represented by the above general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,
R1is selected from
Figure GPA0000234146600000062
R1aSelected from H, C2-4Alkenyl radical, C2-4Alkynyl, C3-7Carbocyclyl, 3-to 6-membered heterocyclyl, C3-7carbocyclyl-C1-4Alkylene or 3-to 6-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CF3、OH、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1, 2 or 3 heteroatoms selected from N, O or S;
R1bis selected from C2-4Alkenyl radical, C2-4Alkynyl, C3-7Carbocyclyl, 3-to 6-membered heterocyclyl, C3-7carbocyclyl-C1-4Alkylene or 3-to 6-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CF3、OH、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1, 2 or 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy, cyano, NH2、C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylthio or-C (═ O) NH2NH of said2、-C(＝O)NH2Or alkyl is optionally further substituted by 0, 1 or 2 substituents selected from F, Cl, Br, I, hydroxy, cyano or C1-4Alkyl is substituted by a substituent;
ring A and ring C are each independently selected from a thiophene ring, furan ring, pyridine ring or benzene ring, preferably a benzene ring, said thiophene ring, furan ring, pyridine ring or benzene ring being optionally further substituted with 0, 1, 2, 3 or 4 groups selected from F, Cl, Br, I, C1-4Alkyl or C1-4The substituent of the alkoxy group is preferably further substituted by 0, 1,2.3 or 4 substituents selected from F, Cl, Br, I, methyl, ethyl, methoxy or ethoxy;
m is selected from the group consisting of a bond, -CH2-、-CH2CH2-、-O-、-S-、-OCH2-、-SCH2-or-CH ═ CH-, preferably selected from the group consisting of bonds, -O-or-S-;
R1dselected from H, hydroxy, -CH2OH or C1-4Alkyl, preferably selected from H, hydroxy, -CH2OH, methyl, ethyl or propyl;
R1eand R1fEach independently selected from F, Cl, Br, I, CF3、NH2OH, cyano, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylthio, -NHC1-4Alkyl or-N (C)1-4Alkyl radical)2；
R2Selected from the group consisting of a bond, C1-6Alkylene radical, C2-6Alkenylene or C2-6Alkynylene, said alkylene, alkenylene or alkynylene being optionally further substituted by 0, 1, 2, 3, 4 or 5 groups selected from F, Cl, Br, I, OH, cyano, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4Substituted by a substituent of alkylene; r2Preferably selected from a bond or C1-6Alkylene optionally further substituted by 0, 1, 2, 3, 4 or 5 groups selected from F, Cl, Br, I, OH, cyano, C1-4Alkyl or C1-4Substituted by a substituent of alkoxy;
R3selected from the group consisting of a bond and C1-6Alkylene optionally further substituted by 0, 1, 2, 3, 4 or 5 groups selected from R3aSubstituted with the substituent(s); r3Preferably selected from a bond or C1-6Alkylene optionally further substituted by 0, 1, 2, 3, 4 or 5 groups selected from R3aSubstituted with the substituent(s);
l is selected from a bond or
Figure GPA0000234146600000071
And Y is directly connected to R2Connecting;
and R is2L and R3The three can not be simultaneously keys;
R4、R5each independently selected from H or C1-4An alkyl group;
b is selected from
Figure GPA0000234146600000072
Figure GPA0000234146600000073
Wherein Q is selected from-CH ═ CH-, -CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-；
R3aSelected from F, Cl, Br, I, cyano, OH, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4An alkylene group; r3aPreferably selected from F, Cl, Br, I, cyano, OH, C1-4Alkyl radical, C1-4Alkoxy or phenyl;
alternatively, two R3aMay form, together with the atoms to which they are attached, a 3 to 6 membered carbocyclic ring optionally further substituted by 0, 1, 2, 3, 4 or 5 substituents selected from F, Cl, Br, I, cyano, OH, C1-4Alkyl or C1-4Substituted by a substituent of alkoxy;
A1and A4Each independently selected from C3-7Cycloalkylene radical, C6-10Arylene, 5-to 8-membered heteroarylene, -O-C6-10Arylene, -O-5 to 8 membered heteroarylene, 5 to 8 membered heteroarylene-O-or C6-10arylene-O-, preferably selected from cyclopentylene, cyclohexylene, phenylene, thienylene, furylene, thiazolylene, oxazolylene, imidazolylene or pyridylene wherein said cycloalkylene, arylene, heteroarylene, cyclopentylene, cyclohexylene, phenylene, thienylene, furylene, thiazolylene, oxazolylene, imidazolylene or pyridylene is optionally further substituted with 0, 1, 2, 3, 4 or 5 groups selected from R6Substituted with the substituent(s);
A2and A3Each independently selected from C1-6Alkylene, or a mixture thereof,C2-6Alkenylene or C2-6Alkynylene, preferably selected from C1-6Alkylene, more preferably selected from methylene, ethylene, propylene, butylene or pentylene, said alkylene, alkenylene, alkynylene or methylene, ethylene, propylene, butylene or pentylene being optionally further substituted by 0, 1, 2, 3, 4 or 5 groups selected from F, Cl, Br, I, OH, cyano, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4Substituted by a substituent of alkylene;
x and Y are each independently selected from the group consisting of a bond, -O-, -C (═ O) O-, -OC (═ O) -, -S (═ O)2-、-C(＝O)NR7-、-NR7C(＝O)-、-OC(＝O)NR7-、-NR7C(＝O)O-、-NR7C(＝O)NR7、-NR7S(＝O)2-、-S(＝O)2NR7-、-NR7S(＝O)2NR7or-NR7-；
R6Each independently selected from F, Cl, Br, I, OH, NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C2-4Alkynyl, C1-4Alkoxy, -OC3-6Cycloalkyl radical, C1-4Alkylthio, -S (═ O) -C1-4Alkyl, -S (═ O)2-C1-4Alkyl, -C (═ O) -C1-4Alkyl, 5-to 6-membered heteroaryl or-C (═ O) O-C1-4Alkyl, alkynyl, alkoxy, cycloalkyl, heteroaryl or NH2Optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CF3、C1-4Alkyl radical, C1-4Alkoxy or-C (═ O) -C1-4Substituted by alkyl substituents, R6Preferably selected from F, Cl, Br, cyano, methyl, ethyl, propyl, isopropyl, CHF2、CF3Methoxy, ethoxy, -OCHF2、-OCF3Cyclopropyloxy, ethynyl, propynyl, pyrrolyl, imidazolyl, pyrazolyl, thienyl, furyl, thiazolyl or oxazolyl;
R7is selected from H or C1-4Alkyl, preferably selected from H, methyl or ethyl;
a is 0, 1, 2, 3 or 4;
b is 0, 1, 2, 3, 4 or 5;
m, n, p or q are each independently selected from 0 or 1.
In a preferred embodiment of the present invention, in the compound represented by the above general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,
R1is selected from
Figure GPA0000234146600000081
R1aSelected from H, ethenyl, ethynyl, propynyl, phenyl, cyclopropylalkyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, benzyl, thienyl, furyl or pyridyl, preferably selected from ethenyl, ethynyl, phenyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, benzyl or thienyl, said ethenyl, ethynyl, propynyl, phenyl, cyclopropylalkyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, benzyl, thienyl, furyl or pyridyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CF3、OH、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy or-O (═ O) C1-4Alkyl is substituted by a substituent;
R1bselected from the group consisting of vinyl, ethynyl, propynyl, phenyl, cyclopropylalkyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, benzyl, thienyl, furyl or pyridyl, preferably selected from the group consisting of vinyl, ethynyl, phenyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, benzyl or thienyl, said vinyl, ethynyl, propynyl, phenyl, cyclopropylalkyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, benzyl, thienyl, furyl or pyridyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from the group consisting of F, Cl, Br, I, CF3、OH、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy or-O (═ O) C1-4Alkyl is substituted by a substituent;
R1cselected from H, hydroxy, NH2Methyl, ethyl, cyano, methylamino, dimethylamino, methoxy, ethoxy, -CH2OH or-C (═ O) NH2Preferably selected from H, hydroxy, NH2Methyl, ethyl or methoxy;
m is selected from the group consisting of a bond, -CH2-、-CH2CH2-、-O-、-S-、-OCH2-、-SCH2-or-CH ═ CH-, preferably selected from the group consisting of bonds, -O-or-S-;
R1dselected from H, NH2Hydroxy, -CH2OH, methyl or ethyl;
R1eand R1fEach independently selected from F, Cl, Br, I, CF3、NH2OH, cyano, methyl, ethyl, methoxy, ethoxy, -NHCH3or-N (CH)3)2；
R2Selected from the group consisting of a bond and C1-6Alkylene, preferably selected from the group consisting of a bond, methylene, ethylene, propylene, butylene or pentylene, said C1-6Alkylene, methylene, ethylene, propylene, butylene or pentylene optionally further substituted with 0, 1, 2, 3, 4 or 5 substituents selected from F, Cl, Br, I, cyano, OH, methyl, ethyl, methoxy or ethoxy;
R3selected from the group consisting of a bond and C1-6Alkylene, preferably selected from the group consisting of a bond, methylene, ethylene, propylene, butylene, pentylene or
Figure GPA0000234146600000091
Said C1-6Alkylene, methylene, ethylene, propylene, butylene, pentylene, or
Figure GPA0000234146600000092
Optionally further substituted with 0, 1, 2, 3, 4 or 5 substituents selected from F, Cl, Br, I, cyano, OH, methyl, ethyl, methoxy or ethoxy;
l is selected from a bond or
Figure GPA0000234146600000093
And Y is directly connected to R2Connecting;
and R is2L and R3The three can not be simultaneously keys;
R4、R5each independently selected from H, methyl or ethyl;
b is selected from
Figure GPA0000234146600000101
Figure GPA0000234146600000102
X and Y are each independently selected from the group consisting of a bond, -O-, -C (═ O) NR7-、-NR7C(＝O)-、-OC(＝O)NR7-or-NR7C(＝O)O-；
A1And A4Each independently selected from the group consisting of phenylene, cyclopentylene, cyclohexylene, phenylene, thienylene, furylene, thiazolylene, oxazolylene, imidazolylene and pyridylene, preferably from the group consisting of phenylene, thienylene, furylene and pyridylene, said phenylene, cyclopentylene, cyclohexylene, phenylene, thienylene, furylene, thiazolylene, oxazolylene, imidazolylene and pyridylene being optionally further substituted with 0, 1, 2, 3 or 4 groups selected from F, Cl, Br, cyano, methyl, ethyl, propyl, isopropyl, CHF2、CF3Methoxy, ethoxy, -OCHF2、-OCF3Ethynyl, propynyl, pyrrolyl, imidazolyl, pyrazolyl, thienyl, furyl, thiazolyl, or oxazolyl;
A2and A3Each independently selected from methylene, ethylene, propylene, butylene or pentylene;
R7is selected from H or C1-4Alkyl, preferably selected from H, methyl, ethyl or propyl;
a is 0, 1, 2, 3 or 4, preferably 0 or 1;
b is 0, 1, 2, 3 or 4, preferably 0, 1 or 2;
m, n, p or q are each independently selected from 0 or 1.
In a preferred embodiment of the present invention, in the compound represented by the above general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,
R1is selected from
Figure GPA0000234146600000103
Figure GPA0000234146600000111
R2Selected from the group consisting of a bond, methylene, ethylene, propylene, butylene or pentylene, said methylene, ethylene, propylene, butylene or pentylene being optionally further substituted with 0, 1, 2, 3, 4 or 5 substituents selected from the group consisting of F, Cl, Br, I, cyano, OH, methyl, ethyl, methoxy or ethoxy;
R2preferably selected from the group consisting of a bond, methylene, ethylene, propylene, -CH2CH(CH3)-、-CH2C(CH3)2-、-C(CH3)2CH2-、-CH(CH3)CH2-, butylene, -CH (CH)3)CH2CH2-、-CH2CH(CH3)CH2-、-CH2CH(CH3)CH2-or pentylene;
R3selected from a bond, methylene, ethylene, propylene, butylene, pentylene or
Figure GPA0000234146600000112
The methylene, ethylene, propylene, butylene, pentylene or
Figure GPA0000234146600000113
Optionally further substituted with 0, 1, 2, 3, 4 or 5 substituents selected from F, Cl, Br, I, cyano, OH, methyl, ethyl, methoxy or ethoxy;
R3preferably selected from the group consisting of a bond, methylene, ethylene, propylene, -CH2CH(CH3)-、-CH2C(CH3)2-、-C(CH3)2CH2-、-CH(CH3)CH2-, butylene, -CH (CH)3)CH2CH2-、-CH2CH(CH3)CH2-、-CH2CH(CH3)CH2-, pentylene or
Figure GPA0000234146600000114
L is selected from
Figure GPA0000234146600000115
And Y is directly connected to R2Connecting;
and R is2L and R3The three can not be simultaneously keys;
b is selected from
Figure GPA0000234146600000116
Figure GPA0000234146600000117
R4、R5Each independently selected from H, methyl or ethyl;
x and Y are each independently selected from the group consisting of a bond, -O-, -C (═ O) NR7-、-NR7C(＝O)-、-OC(＝O)NR7-or-NR7C(＝O)O-；
A1And A4Each independently selected from phenylene, thienylene, furanylene or pyridylene, preferably from phenylene, said phenylene, thienylene, furanylene or pyridylene being optionally further substituted by 0, 1, 2, 3 or 4 groups optionally selected from F, Cl, Br, CHF2、CF3Cyano, methyl, ethyl, methoxy, ethoxy, -OCHF2、-OCF3Ethynyl, propynyl, pyrrolyl, imidazolyl or pyrazolyl;
A2and A3Each independently selected from methylene, ethylene, propylene, butylene or pentylene;
R7is selected from H or C1-4Alkyl, preferably selected from H, methyl, ethyl or propyl;
m, n, p or q are each independently selected from 0 or 1.
In a preferred embodiment of the present invention, in the compound represented by the above general formula (I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, the compound includes, but is not limited to:
Figure GPA0000234146600000131
Figure GPA0000234146600000141
the invention also provides a pharmaceutical composition, which contains a therapeutically effective dose of the compound of the general formula (I) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, and a pharmaceutically acceptable carrier, a diluent, an adjuvant, a vehicle or an excipient, and can further comprise one or more other therapeutic agents, preferably, the other therapeutic agents are selected from one or more of a PDE4 inhibitor, a muscarinic receptor antagonist, a corticosteroid and a β -adrenergic receptor agonist.
The invention also provides the compound of the general formula (I) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, and application of the compound in preparing a medicament for treating airway obstructive diseases, preferably application in preparing a medicament for treating asthma, chronic obstructive pulmonary disease or bronchitis.
The present invention also provides a method for treating an obstructive airways disease, which comprises administering a compound represented by the general formula (I) above or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a co-crystal or a prodrug thereof, or a pharmaceutical composition thereof.
The present invention also provides a method of treating asthma, chronic obstructive pulmonary disease or bronchitis, which comprises administering a compound of the general formula (I) as described above, or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, or a pharmaceutical composition as described above.
The invention also provides an intermediate for preparing the compound shown in the general formula (I) or the stereoisomer thereof, wherein the intermediate is selected from the compound shown in the general formula (II) or the stereoisomer thereof:
Figure GPA0000234146600000142
wherein:
R1is selected from
Figure GPA0000234146600000151
R1aSelected from H, C2-6Alkenyl radical, C2-6Alkynyl, C3-10Carbocyclyl, 3-to 8-membered heterocyclyl, C3-10carbocyclyl-C1-4Alkylene or 3-to 8-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1 to 3 heteroatoms selected from N, O or S;
R1bis selected from C2-6Alkenyl radical, C2-6Alkynyl, C3-10Carbocyclyl, 3-to 8-membered heterocyclyl, C3-10carbocyclyl-C1-4Alkylene or 3-to 8-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0, 1, 2, 3 or 4 substituents selected from F, Cl, Br、I、CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1 to 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy, cyano, NH2、C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio or-C (═ O) NH2NH of said2、-C(＝O)NH2Or alkyl is optionally further substituted by 0 to 2 substituents selected from F, Cl, Br, I, hydroxy, cyano or C1-4Alkyl is substituted by a substituent;
R1eand R1fEach independently selected from F, Cl, Br, I, CF3、NH2OH, carboxyl, cyano, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylthio, -NHC1-4Alkyl, -N (C)1-4Alkyl radical)2、-S(＝O)-C1-4Alkyl, -S (═ O)2-C1-4Alkyl or-C (═ O) O-C1-4An alkyl group;
R9selected from H or an amino protecting group;
a is 0, 1, 2, 3 or 4;
b is 0, 1, 2, 3, 4 or 5.
In a preferred embodiment of the present invention, in the compound represented by the above general formula (II) or a stereoisomer thereof,
R1is selected from
Figure GPA0000234146600000152
Figure GPA0000234146600000161
R1Preferably selected from
Figure GPA0000234146600000162
R9Selected from H or an amino protecting group, wherein said amino protecting group is preferably benzyl, p-methoxybenzyl, tert-butoxycarbonyl, benzyloxycarbonyl, acetyl or benzoyl.
In a preferred embodiment of the present invention, the intermediate represented by the above general formula (II) for preparing the compound represented by the general formula (I) or a stereoisomer thereof is selected from one of the following compounds:
Figure GPA0000234146600000163
the invention relates to an intermediate for preparing a compound shown as a general formula (I) or a stereoisomer thereof, wherein the intermediate is selected from a compound shown as a general formula (III) or a stereoisomer thereof:
Figure GPA0000234146600000164
R1is selected from
Figure GPA0000234146600000165
R1aSelected from H, C2-6Alkenyl radical, C2-6Alkynyl, C3-10Carbocyclyl, 3-to 8-membered heterocyclyl, C3-10carbocyclyl-C1-4Alkylene or 3-to 8-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1 to 3 heteroatoms selected from N, O or S;
R1bis selected from C2-6Alkenyl radical, C2-6Alkynyl, C3-10Carbocyclyl, 3 to8-membered heterocyclic group, C3-10carbocyclyl-C1-4Alkylene or 3-to 8-membered heterocyclyl-C1-4Alkylene, said alkenyl, alkynyl, alkylene, carbocyclyl or heterocyclyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3、NH2Carboxy, cyano, nitro, C1-4Alkyl radical, C1-4Alkoxy, -O (═ O) C1-4Alkyl, - (═ O) C1-4Alkyl, -OC3-6Cycloalkyl or C1-4Alkylthio containing 1 to 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy, cyano, NH2、C1-6Alkyl radical, C1-6Alkoxy radical, C1-6Alkylthio or-C (═ O) NH2NH of said2、-C(＝O)NH2Or alkyl is optionally further substituted by 0 to 2 substituents selected from F, Cl, Br, I, hydroxy, cyano or C1-4Alkyl is substituted by a substituent;
R1eand R1fEach independently selected from F, Cl, Br, I, CF3、NH2OH, carboxyl, cyano, C1-4Alkyl radical, C1-4Alkoxy radical, C1-4Alkylthio, -NHC1-4Alkyl, -N (C)1-4Alkyl radical)2、-S(＝O)-C1-4Alkyl, -S (═ O)2-C1-4Alkyl or-C (═ O) O-C1-4An alkyl group;
R2selected from the group consisting of a bond, C1-6Alkylene radical, C2-6Alkenylene or C2-6Alkynylene, said alkylene, alkenylene or alkynylene being optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I, OH, cyano, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4Substituted by a substituent of alkylene;
w is selected from OH, carboxy, -C (═ O) OC1-4Alkyl, phenyl, -C (═ O) NRx-phenyl, -ONRxC (═ O) -phenyl, -C (═ O) NRx-C1-4alkylene-C (═ O) OH, -C (═ O) NRx-C1-4alkylene-C (═ C)O)OC1-4Alkyl, -C (═ O) NRx-C1-4alkylene-C (═ O) NRx-phenyl, -C (═ O) NRx-phenylene-C (═ O) NRx-phenyl,
Figure GPA0000234146600000171
Said phenyl group is optionally further substituted with 0 to 4 substituents selected from the group consisting of F, Cl, Br, I, methoxy, ethoxy,
Figure GPA0000234146600000172
Formyl, carboxy, -C (═ O) OC1-4Alkyl, aryl, heteroaryl, and heteroaryl,
Figure GPA0000234146600000173
Substituted with the substituent(s);
Rxis selected from H or C1-4An alkyl group;
R3selected from the group consisting of a bond and C1-6Alkylene optionally further substituted with 0 to 5 substituents selected from R3aIs substituted by a substituent of (A), R3aSelected from F, Cl, Br, I, cyano, OH, C1-4Alkyl radical, C1-4Alkoxy, phenyl or phenyl-C1-4An alkylene group; alternatively, two R3aMay form, together with the atoms to which they are attached, a 3 to 6 membered carbocyclic ring optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I, cyano, OH, C1-4Alkyl or C1-4Substituted by a substituent of alkoxy;
b is selected from
Figure GPA0000234146600000181
Figure GPA0000234146600000182
Wherein Q is selected from-CH ═ CH-, -CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-；
B is preferably selected from
Figure GPA0000234146600000183
Figure GPA0000234146600000184
a is 0, 1, 2, 3 or 4;
b is 0, 1, 2, 3, 4 or 5.
In a preferred embodiment of the present invention, the intermediate for preparing the compound represented by the general formula (I) or a stereoisomer thereof represented by the above general formula (III) is selected from one of the following compounds:
Figure GPA0000234146600000185
Figure GPA0000234146600000191
Figure GPA0000234146600000201
the ditrifluoroacetate salt in the compound of the embodiment of the present invention can be dissolved in a polar organic solvent (e.g., a mixed solvent of methanol and dichloromethane (v/v. 1/90)), the pH is adjusted to be alkaline by adding an alkaline reagent (e.g., a saturated sodium bicarbonate solution or a saturated sodium carbonate solution), the mixture is stirred and extracted with an organic solvent (e.g., dichloromethane, ethyl acetate, etc.), and the free base form of the corresponding compound can be obtained by concentrating the organic phase under reduced pressure.
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 group may optionally be further substituted by 0, 1, 2, 3, 4 or 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)a-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, R as appearing herein19And R19aAs defined above.
"alkylene group"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 may optionally be further substituted by 0, 1, 2, 3, 4 or 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)a-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 herein defined and defined
"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-carbon double bonds, and the main chain includes 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms, and still more preferably 2 to 4 carbon atoms in the main chain, and 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-hexenylAlkenyl, 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, 1, 2, 3, 4 or 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)a-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.
"alkenylene" refers to a divalent alkenyl group in which the alkenyl group is as defined 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, 1, 2, 3, 4 or 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)a-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.
"alkynylene" refers to a divalent alkynyl group in which the alkynyl group is as defined 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, 1, 2, 3, 4 or 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)a-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.
"cycloalkylene" refers to a divalent cycloalkyl group, wherein cycloalkyl is as defined above.
"aryl" means a monovalent aromatic hydrocarbon group having a single ring or fused rings, typically 6 to 10 carbon atoms, non-limiting examples of which include phenyl, naphthalen-1-yl or naphthalen-2-yl. Said aryl group may optionally be further substituted by 0, 1, 2, 3, 4 or 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)a-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). Aryl, as found herein, is defined as above.
"arylene" refers to a divalent aromatic radical, wherein aryl is as defined above.
"heteroaryl" refers to a monovalent aryl group having a single ring or two fused rings and containing at least 1 heteroatom selected from N, O or S in the ring, typically consisting of 5 to 8 atoms, non-limiting examples of which include pyrrolyl, imidazolyl, thiazolyl, thienyl, furyl, pyrazolyl, isoxazolyl, oxazolyl, pyridyl, or pyrazinyl. Said heteroaryl may optionally be further substituted by 0, 1, 2, 3, 4 or 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)a-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). Heteroaryl, as found herein, is defined as above.
"heteroarylene" refers to a divalent heteroaryl group, wherein heteroaryl is as defined above.
"carbocyclyl" refers to a saturated or unsaturated aromatic or non-aromatic ring which may be 3 to 10 memberedA monocyclic, 4-to 12-membered bicyclic or 10-to 15-membered tricyclic ring system to which the carbocyclic group may be attached an endocyclic or spirocyclic 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, spirocyclo-, cyclo,
Figure GPA0000234146600000231
Said carbocyclyl may optionally be further substituted by 0, 1, 2, 3, 4 or 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)a-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.
"heterocyclyl" means 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 and which contains 1 to 4 heteroatoms selected from N, O or S, preferably a 3 to 8 membered heterocyclyl group, the optionally substituted N, S ring of which may be oxidized to various oxidation states. The heterocyclic group may be attached at a heteroatom or carbon atom, the heterocyclic group may be attached to a bridged or 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, and oxazepinylDiazepine, thiazepine, pyridyl, piperidyl, homopiperidinyl, furyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, piperidyl, perinyl, morpholinyl, thiomorpholinyl, thiaoxazolidyl, 1, 3-dithianyl, dihydrofuranyl, dihydropyranyl, dithiafuranyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, chromanyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1, 3-dioxolyl, and dihydropyrrolyl, Pyrazolinyl, dithianyl, dithienoalkyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 1, 2, 3, 4-tetrahydroisoquinolinyl, 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, 1, 2, 3, 4 or 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)a-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.
"β -adrenoceptor binding group" refers to a group capable of binding to β -adrenoceptorSee, for example, the review article "β -acquired receivers in Comprehensive Medicinal Chemistry, 1990, B.E.Main, p187(Pergamon Press)". the above groups are also described, for example, in WO/2005092841, US/20050215542, WO/2005070872, WO/2006023460, WO/2006051373, WO/2006087315 and WO/2006032627 non-limiting examples include
Figure GPA0000234146600000241
R4、R5Each independently selected from H or C1-4Alkyl, B is selected from
Figure GPA0000234146600000251
Figure GPA0000234146600000252
Wherein Q is selected from-CH ═ CH-, -CH2CH2-、-O-、-S-、-CH2O-、-OCH2-、-C(CH3)2O-or-OC (CH)3)2-。
"amino protecting group" refers to a group used for amino protection which is suitable for protecting an amino group from chemical reaction, but which is easily removed after the desired chemical reaction is completed in the rest of the molecule. The protection of amino Groups In the protection group In Organic Synthesis (the university of eastern science and technology, the university of Organic chemistry, book of honor, school of eastern science and technology, published by the university of eastern science and technology, 2004; original book Protective Groups In Organic Synthesis (third edition), Theodora W.Green and Peter G.M.Wuts) is described In detail In the chapter on amino protection. Page 494-653 of protective groups in organic synthesis is incorporated herein by reference as part of the present application.
"amino protecting group" includes, but is not limited to, the following groups: benzyl, p-methoxybenzyl, trityl, tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, 2, 2, 2-trichloroethoxycarbonyl, ethoxycarbonyl, benzyloxycarbonyl, trifluoroacetyl, acetyl or benzoyl.
"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 several of the compounds described herein or a physiologically/pharmaceutically acceptable salt thereof with other constituents, wherein the other constituents 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 () at 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).
HP L C was determined using an Agilent 1260DAD high pressure liquid chromatograph (Zorbax SB-C18100 × 4.6.6 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 the thin layer chromatography (T L C) 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 of 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, Shaoshan far chemical technology, and Bailingwei technology.
Nitrogen atmosphere means that the reaction flask is connected to a nitrogen balloon of about 1L volume.
The hydrogen atmosphere refers to a reaction flask with a hydrogen balloon attached to it of about 1L volume.
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 ℃.
TBS is tert-butyl dimethyl silicon base.
Boc is tert-butyloxycarbonyl.
TFA is trifluoroacetic acid.
HATU is 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (CAS: 148893-10-1).
Example 1: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxyanilino ] -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] -2-hydroxy-2, 2-bis (2-thienyl) acetate (Compound 1)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000271
The first step is as follows: 2-hydroxy-7-azaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (1B)
tert-butyl 2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate
Figure GPA0000234146600000272
Sodium borohydride (1.1g, 29.3mmol) was added to a solution of tert-butyl 2-carbonyl-7-azaspiro [3.5] nonane-7-carboxylate (1A) (7g, 29.3mmol) in methanol (100m L) under ice-bath, after completion of addition, reaction was carried out at room temperature for 2 hours, water (100m L) was added, extraction was carried out with dichloromethane (100m L× 3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the title compound, tert-butyl 2-hydroxy-7-azaspiro [3.5] nonane-7-carboxylate (1B), as a white solid (7g, 99.2% yield).
1H NMR(400MHz，CDCl3)4.39-4.23(m，1H)，3.38-3.22(m，4H)，2.32-2.22(m，2H)，2.04(d，1H)，1.74-1.62(m，2H)，1.50(m，4H)，1.45-1.41(m，9H).
LCMS m/z＝264.1[M+23].
The second step is that: 2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (1C)
tert-butyl 2-[2-hydroxy-2，2-bis(2-thienyl)acetyl]oxy-7-azaspiro[3.5]nonane-7-carboxylate
Figure GPA0000234146600000281
Tert-butyl 2-hydroxy-7-azaspiro [3.5] nonane-7-carboxylate (1B) (0.482g, 2mmol) was dissolved in toluene (10m L), sodium hydride (0.024g, 1mmol) was added, stirring was carried out for 5 minutes, methyl 2-hydroxy-2, 2-bis (2-thienyl) acetate (0.508g, 2mmol) was added, the reaction was warmed to 155 ℃ and quenched with water, successively extracted with ethyl acetate (20m L× 2), washed with a saturated aqueous sodium chloride solution (20m L× 1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent ethyl acetate/petroleum ether (v: v) ═ 1/100 to 1/20) to give the title compound 2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] nonane-7-carboxylate (1C) as a pale yellow solid (yield, 0.23g, 25%).
1H NMR(400MHz，CDCl3)7.29(dd，2H)，7.17(dd，2H)，6.98(dd，2H)，5.25-5.05(m，1H)，4.66(s，1H)，3.32(dd，2H)，3.29-3.23(m，2H)，2.41-2.30(m，2H)，1.88(dd，2H)，1.54-1.50(m，2H)，1.47(d，2H)，1.44(s，9H).
LCMS m/z＝486.3[M+23].
The third step: 7-Azaspiro [3.5] nonan-2-yl 2-hydroxy-2, 2-bis (2-thienyl) acetate (1D)
7-azaspiro[3.5]nonan-2-yl 2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000282
Tert-butyl 2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] nonane-7-carboxylate (1C) (0.5g, 1.08mmol) was dissolved in 1, 4-dioxane (10m L), excess hydrochloric acid gas was bubbled through, and reacted at room temperature for 2 hours, water (20m L) was added, pH adjusted to 8 to 9 with solid potassium carbonate at 0 ℃, extracted with ethyl acetate (20m L× 2), washed with saturated aqueous sodium chloride (20m L× 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 7-azaspiro [3.5] nonane-2-yl 2-hydroxy-2, 2-bis (2-thienyl) acetate (1D), as a white solid (0.24g, 61% yield).
1H NMR(400MHz，CDCl3)7.30(dd，2H)，7.16(dd，2H)，6.98(dd，2H)，5.15(t，1H)，4.61(s，1H)，3.08(s，2H)，3.01(s，2H)，2.45-2.33(m，2H)，1.99-1.87(m，4H)，1.87-1.79(m，2H).
LCMS m/z＝364.0[M+1].
The fourth step: [7- [3- (2-chloro-4-formyl-5-methoxyanilino) -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] -2-hydroxy-2, 2-bis (2-thienyl) acetate (1F)
[7-[3-(2-chloro-4-formyl-5-methoxy-anilino)-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-y1]-2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000291
7-azaspiro [3.5] nonane-2-yl 2-hydroxy-2, 2-bis (2-thienyl) acetate (1D) (2.6g, 7.153mmol) and N- (2-chloro-4-formyl-5-methoxyphenyl) propan-2-enamide (1E) (prepared by reference to the synthetic example in WO2010119064a 1) (1.886g, 7.868mmol) were suspended in 2-methyltetrahydrofuran (15m L), triethylamine (1.448g, 14.31mmol) was added, the reaction was stopped at 100 ℃ with a microwave for 30 minutes, the reaction liquid was concentrated under reduced pressure after cooling to room temperature, and the residue was purified by silica gel column chromatography (eluent dichloromethane/methanol (v: v) ═ 100/1 to 20/1) to give the title compound [7- [3- (2-chloro-4-formyl-5-methoxyanilino) -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] -2-hydroxy-2-acetate (1g, 2-thienyl) as a pale yellow solid (46%).
1H NMR(400MHz，CDCl3)10.94(s，1H)，10.30(s，1H)，8.34(s，1H)，7.81(s，1H)，7.29(dd，2H)，7.18(dd，2H)，6.98(dd，2H)，5.16(s，1H)，4.67(s，1H)，3.93(s，3H)，3.78(s，1H)，2.63(s，4H)，2.46-2.31(m，4H)，1.90(dd，2H)，1.68(d，4H).
LCMS m/z＝603.0[M+1].
The fifth step: [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (1H)
[7-[3-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-2-chloro-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000301
[7- [3- (2-chloro-4-formyl-5-methoxyanilino) -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] -2-hydroxy-2, 2-bis (2-thienyl) acetate (1F) (0.603G, 1mmol) and 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.401G, 1.2mmol) were dissolved in a mixed solvent of dichloromethane (5m L) and methanol (5m L), stirred at room temperature for 30 minutes, sodium triacetoxyborohydride (0.636G, 3.0mmol) was added, reacted at room temperature for 2 hours, dichloromethane (20m L) was added to the reaction solution, and then washed with a saturated sodium bicarbonate solution (20m L× 2) and a saturated sodium chloride aqueous solution (20m L×) in order, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified as a silica gel column (silica-3-ethyl-2-hydroxy-2-phenyl) -7-azaspiro [ 3.5-hydroxy-2-yl ] quinoline (1G, 1.6-bis (7-hydroxy-2-silica-2-ethyl-2-ethyl-2-one half) was added to obtain the title compound, which was purified as a pale yellow solid.
And a sixth step: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxyanilino ] -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (Compound 1)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000302
[7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (1H) (0.5g, 0.543mmol) was dissolved in dichloromethane (10m L), triethylamine trihydrofluoride (5m L) was added, reaction was carried out at 35 ℃ for 6 hours, dichloromethane (50m L) was added to the reaction solution, pH was adjusted to about 9 with a saturated sodium bicarbonate solution, dichloromethane (30m L×) was then extracted, the organic phases were combined, washed with a saturated chlorinated solution (30m L×), anhydrous sodium sulfate was dried, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 2 v) ═ 2-hydroxy-2-oxo-2-hydroxy-2, 2-bis (7-thienyl-2-ethyl) -2, 2-bis (7-2-thienyl-2, 2-bis (7-2-thienyl) ethyl-2, 2-bis (7-2-bis (9) ethyl) yield).
1H NMR(400MHz，DMSO-d6)10.30(s，1H)，8.12(d，1H)，7.75(s，1H)，7.47(dd，2H)，7.30(s，2H)，7.09(dd，2H)，7.05(d，1H)，6.99(dd，2H)，6.91(d，1H)，6.46(d，1H)，5.11-5.00(m，2H)，3.71(s，3H)，3.64(s，2H)，2.67(dd，2H)，2.57(d，2H)，2.53(d，2H)，2.43-2.14(m，6H)，1.76(dd，2H)，1.60(s，2H)，1.55(s，2H).
LCMS m/z＝807.1[M+1].
Example 2: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] non-2-yl ] N- (2-phenylphenyl) carbamate (Compound 2)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000311
Figure GPA0000234146600000321
The first step is as follows: 1-isocyanate-2-phenyl-benzene (2B)
1-isocyanato-2-phenyl-benzene
Figure GPA0000234146600000322
2-aminobiphenyl (2A) (2.4g, 14mmol) was dissolved in toluene (50m L), triphosgene (1.7g, 5.7mmol) was added, and the reaction was reacted at 120 ℃ for 1 hour, after the reaction solution was cooled to room temperature, concentrated under reduced pressure to give the title compound 1-isocyanate-2-phenyl-benzene (2B) as a brown oil (2.7g, 98% yield).
The second step is that: 2- [ (2-Phenylphenyl) carbamoyloxy ] -7-azaspiro [3, 5] nonane-7-aminocarboxylic acid tert-butyl ester (2C)
tert-butyl 2-[(2-phenylphenyl)carbamoyloxy]-7-azaspiro[3.5]nonane-7-carboxylate
Figure GPA0000234146600000323
1-isocyanate-2-phenyl-benzene (2B) (2.7g, 13.4mmol) was dissolved in tetrahydrofuran (50m L), 2-hydroxy-7-azaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (1B) (1.9g, 7.87mmol) was added, triethylamine (2.39g, 23.6mmol) was added, and the reaction mixture was reacted at 70 ℃ for 4 hours, after cooling to room temperature and concentrating under reduced pressure, the residue was purified by silica gel column chromatography (eluent ethyl acetate: petroleum ether (v/v) ═ 0: 1 to 1: 9) to give the title compound, tert-butyl 2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3, 5] nonane-7-aminocarboxylate (2C), as a yellow oil (3.4g, 99% yield).
LCMS m/z＝459.2[M+23].
The third step: 7-Azaspiro [3, 5] nonan-2-yl N- (2-phenylphenyl) carbamate (2D)
7-azaspiro[3.5]nonan-2-yl N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000331
2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3, 5] nonane-7-aminocarboxylic acid tert-butyl ester (2C) (3.4g, 7.8mmol) was dissolved in dichloromethane (15m L), trifluoroacetic acid (4.4g, 39mmol) was added, reaction was carried out at room temperature for 3 hours, the reaction mixture was concentrated under reduced pressure, then water (20m L) and dichloromethane (20m L) were added, aqueous ammonia was added to adjust pH to 9, extraction was carried out, the aqueous phase was extracted with dichloromethane (20m L× 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 (ethyl acetate: petroleum ether (v/v) ═ 1: 4 to 1: 0; methanol: dichloromethane (v/v) ═ 1: 9) to give the title compound 7-azaspiro [3, 5] nonan-2-yl N- (2-phenylphenyl) carbamate (2D) as a yellow solid (2.1g, 80% yield).
1H NMR(400MHz，CDCl3)8.11(d，1H)，7.5(dd，2H)，7.44-7.38(m，1H)，7.38-7.31(m，3H)，720(dd，1H)，7.15-7.07(m，1H)，6.66(s，1H)，5.08-4.90(m，1H)，2.81-2.75(m，2H)，2.72(t，3H)，2.37-2.24(m，2H)，1.85-1.74(m，2H)，1.60-1.50(m，4H).
LCMS m/z＝337.1[M+1].
The fourth step: [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3, 5] nonan-2-yl N- (2-phenylphenyl) carbamate (2E)
[7-[3-(2-chloro-4-formyl-5-methoxy-anilino)-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000332
N- (2-chloro-4-formyl-5-methoxyphenyl) acrylamide (1E) (0.648g, 2.71mmol) was dissolved in 2-methyltetrahydrofuran (10m L), 7-azaspiro [3, 5] nonan-2-yl N- (2-phenylphenyl) carbamate (2D) (0.700g, 2.08mmol) was added, acetic acid (0.250g, 4.16mmol) was added, microwave reaction was performed at 100 ℃ for 1 hour, the reaction solution was concentrated under reduced pressure, methylene chloride (20m L) was added to the residue, saturated aqueous sodium bicarbonate solution (20m L) was added, the aqueous phase was extracted with (20m L× 2) eluent, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v/v) ═ 1: 1 to 1: 0), and the title compound [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3- (2-phenyl) carbamate (yield, N-azaspiro [ 2-phenyl) was obtained as a white solid (2g, 2.83%).
1H NMR(400MHz，CDCl3)10.30(s，1H)，8.33(s，1H)，8.11(d，1H)，7.81(s，1H)，7.50(dd，2H)，7.45-7.39(m，1H)，7.39-7.33(m，3H)，7.21(dd，1H)，7.15-7.10(m，1H)，6.62(s，1H)，5.06-4.95(m，1H)，3.93(s，3H)，2.83-2.20(m，10H)，1.90-1.77(m，2H)，1.71(s，4H).
LCMS m/z＝576.1[M+1].
The fifth step: [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3, 5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (2F)
[7-[3-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-2-chloro-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000341
Dissolving [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3, 5] nonan-2-yl N- (2-phenylphenyl) carbamate (2E) (1.00G, 1.74mmol) in dichloromethane (10m L) and methanol (10m L), adding 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.581G, 1.74mmol), reacting at room temperature for 1 hour, adding sodium triacetoxyborohydride (1.11G, 5.21mmol), reacting at room temperature for 2 hours, adding dichloromethane (20m L) to the reaction solution, adding a saturated sodium bicarbonate solution (20m L), extracting the aqueous phase with (20m L× 1), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, eluting the residue with silica gel column chromatography (ethyl acetate: 1 v-5-methoxy-anilino-2-phenyl) as an eluant (1.5G, 1.7-azaspiro [ 3-2-yl ] N- (2-phenyl) carbamate (1G, 1.74mmol), purifying the title compound by ethyl-2-amino-1-2-phenyl-1H-2-ethyl-7-phenyl-1-ethyl-7-azaspiro [ 7-8-phenyl ] carba- [ 3-7-azaspiro [ 7-1G, 7-azaspiro [ 7-ethyl ] as a yellow solid.
LCMS m/z＝447.6[(M+2)/2].
And a sixth step: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3, 5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (Compound 2)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000351
Dissolving [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3 oxo-propyl ] -7-azaspiro [3, 5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (2F) (0.900g, 1.01mmol) in dichloromethane (8m L), adding triethylamine trihydrofluoride (1.62g, 10.1mmol), reacting at room temperature for 24 hours, adding water (20m L) and dichloromethane (20m L) to the reaction solution, adding 3% sodium hydroxide solution to adjust pH to about 12, extracting the aqueous phase with (20m L× 2), combining the organic phases, washing with saturated sodium chloride aqueous solution (20m L× 1), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, separating the residue with silica gel column chromatography to obtain a yellow ethyl ester of 3-azaspiro [ 3-2-phenyl ] N- (2-phenyl) carbamate (3-oxo-propyl ] -7-azaspiro [ 3-2-phenyl ] carbamate (0.8-8-phenyl) as a yellow eluent, purifying compound (2-ethyl-phenyl-2-nonan-2-yl) as a yellow solid, purifying compound (7-azaspiro [ 7-azaspiro [ 7-8H-8-phenyl) and 8H).
1H NMR(400MHz，DMSO-d6)10.34(s，1H)，8.60(s，1H)，8.13(d，1H)，7.78(s，1H)，7.55-7.17(m，11H)，7.07(d，1H)，6.94(d，1H)，6.48(d，1H)，5.10(s，1H)，4.74(s，1H)，3.73(d，5H)，2.80-2.66(m，2H)，2.64-2.52(m，4H)，2.38(s，4H)，2.14(s，2H)，1.56(s，6H).
LCMS m/z＝390.6[(M+2)/2].
Example 3: [7- [2- [ [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-phenyl ] carbamoyloxy ] ethyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (Compound 3)
[7-[2-[[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-phenyl]carbamoyloxy]ethyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000361
The first step is as follows: [7- (2-hydroxyethyl) -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (3A)
[7-(2-hydroxyethyl)-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000362
7-azaspiro [3.5] nonan-2-yl 2-hydroxy-2, 2-bis (2-thienyl) acetate (1D) (0.500g, 1.38mmol) was dissolved in acetonitrile (15m L), bromoethanol (0.516g, 4.13mmol) and diisopropylethylamine (0.533g, 4.13mmol) were added, and the reaction mixture was reacted at 90 ℃ for 3 hours, after concentration under reduced pressure, the residue was purified by column chromatography on silica gel (eluent ethyl acetate: petroleum ether (v/v) ═ 1: 1 to 1: 0; methanol: dichloromethane (v/v) ═ 3: 97) to give the title compound [7- (2-hydroxyethyl) -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (3A), a yellow solid (0.54g, 96.3% yield).
1H NMR(400MHz，CDCl3)7.30(dd，2H)，7.16(dd，2H)，6.99(dd，2H)，5.19--5.10(m，1H)，4.04(s，2H)，3.50--2.79(m，6H)，2.43(dd，2H)，2.01(m，5H)，1.61-1.42(m，2H).
LCMS m/z＝408.1[M+1]。
The second step is that: [7- [2- [ (2-chloro-4-formyl-5-methoxy-phenyl) carbamoyloxy ] ethyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (3C)
[7-[2-[(2-chloro-4-formyl-5-methoxy-phenyl)carbamoyloxy]ethyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000371
Dissolving 4-amino-5-chloro-2-methoxy-benzaldehyde (3B) (1.48g, 7.95mmol) in toluene (30m L), adding triphosgene (1.18g, 3.97mmol), reacting at 120 ℃ for 2 hours, removing the solvent under reduced pressure to give reaction solution 1, [7- (2-hydroxyethyl) -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (3A) (0.540g, 1.32mmol) in tetrahydrofuran (20m L), adding reaction solution 1, adding triethylamine (0.670g, 6.62mmol), reacting at 70 ℃ for 2 hours, cooling the reaction solution to room temperature, concentrating under reduced pressure, separating and purifying the residue by silica gel column chromatography (eluent methanol: dichloromethane (v/v) ═ 1: 99 to 5: 95) to give the title compound [7- [2- [ (2-chloro-4-formyl-5-methoxy-phenyl) carbamoyl ] nonane ] acetate (2-hydroxy-2, 2-thienyl ] ethyl ] -7- [2- [ (2-chloro-4-formyl-methoxy-phenyl) carbamoyl) acetate (yield 0.1.73 g, 2-hydroxy-2-thienyl ] ethyl ] 7 (0.73%).
1H NMR(400MHz，CDCl3)10.29(s，1H)，8.01(s，1H)，7.82(s，1H)，7.29(dd，2H)，7.17(dd，2H)，6.98(dd，2H)，5.19-5.08(m，1H)，4.49(s，2H)，3.94(s，3H)，3.09-2.49(m，6H)，2.45-2.32(m，2H)，2.01-1.66(m，6H).
LCMS m/z＝619.2[M+1]。
The third step: [7- [2- [ [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-phenyl ] carbamoyloxy ] ethyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (3D)
[7-[2-[[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-2-chloro-5-methoxy-phenyl]carbamoyloxy]ethyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000372
[7- [2- [ (2-chloro-4-formyl-5-methoxy-phenyl) carbamoyloxy ] ethyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (3C) (0.600G, 0.969mmol) was dissolved in dichloromethane (10m L) and methanol (10m L), 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.324G, 0.969mmol) was added, reaction was performed at room temperature for 1 hour, sodium triacetoxyborohydride (0.619G, 2.91mmol) was added, reaction was performed at room temperature for 3 hours, dichloromethane (20m L) was added to the reaction solution, saturated aqueous sodium bicarbonate solution (20m L) was added, extraction was performed with dichloromethane (20m L× 1) for aqueous phase, anhydrous sodium sulfate was dried, concentration was performed, and the residue was purified by silica gel column chromatography (ethyl ether chromatography (ethyl-2-amino-2-yl) -2-hydroxy-2-quinolin-2-one (1G, 0.7-7-azaspiro [ 7-2-hydroxy-2-1H-1- [ [ 9mmol) was obtained as a yellow solid.
1H NMR(400MHz，CDCl3)8.19(d，1H)，7.78(s，1H)，7.28(dd，2H)，7.17(dd，3H)，7.06(s，1H)，7.03-6.92(m，3H)，6.84(d，1H)，6.59(d，1H)，5.22-5.04(m，2H)，4.38-4.22(m，2H)，3.83-3.71(m，5H)，3.00-2.78(m，2H)，2.71(t，2H)，2.46(d，4H)，2.37-2.28(m，2H)，1.86(dd，2H)，1.63(dd，4H)，0.85(s，9H)，0.02(s，3H)，-0.22(s，3H).
LCMS m/z＝469.3[(M+2)/2]。
The fourth step: [7- [2- [ [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-phenyl ] carbamoyloxy ] ethyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (Compound 3)
[7-[2-[[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-phenyl]carbamoyloxy]ethyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000381
[7- [2- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-phenyl ] carbamoyloxy ] ethyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (3D) (0.420g, 0.448mmol) was dissolved in tetrahydrofuran (8m L), triethylamine trihydrofluoride salt (0.722g, 4.48mmol) was added to the reaction solution at room temperature for 24 hours, dichloromethane (50m L) was added to the reaction solution to adjust pH to about 8, saturated sodium bicarbonate solution was added to the extraction solution, dichloromethane (50m L× 1) was used for extraction, the organic phases were combined, washed with saturated sodium chloride aqueous solution (20m L× 1), anhydrous sodium sulfate was dried, filtered, the filtrate was concentrated under reduced pressure, the residue was chromatographed on silica gel column (ethyl quinolineacetate (ethyl-2- [3 g-2-tert-butyl (2H) -2-oxo-5-yl ] amino ] ethyl ] -7-azaspiro [ 3-2, 2-bis (2-thienyl ] ethyl ] -2, 2-bis (2-thienyl) was purified as a yellow solid, 2-bis (2-thienyl) ethyl) compound, 2-bis (2-bis (2-bis (448) at yield, 2-bis (448-2-bis (448) at room temperature for 24 hours.
1H NMR(400MHz，CD3OD)8.25(d，1H)，7.75(s，1H)，7.39(m，3H)，7.25(d，1H)，7.16(dd，2H)，7.05-6.95(m，3H)，6.64(d，1H)，5.34(t，1H)，5.18-5.06(m，1H)，4.37(t，2H)，4.09(s，2H)，3.85(d，3H)，3.10(d，2H)，2.84(t，2H)，2.62(d，4H)，2.42-2.29(m，2H)，1.88(dd，2H)，1.73(t，2H)，1.66(t，2H).
LCMS m/z＝412.3[(M+2)/2]。
Example 4: [7- [2- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (4-hydroxy-2-oxo-3H-1, 3-benzothiazol-7-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (Compound 4)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(4-hydroxy-2-oxo-3H-1，3-benzothiazol-7-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000391
[7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (1F) (0.420g, 0.696mmol) was dissolved in dichloromethane (10m L) and methanol (10m L), 7- [ (1R) -2-amino-1-hydroxy-ethyl ] -4-hydroxy-3H-1, 3-benzothiazol-2-one (4A, reference Bioorganic & Medicinal Chemistry L etters, 21), 4612- & 4616; prepared) (0.2011g, 0.766), sodium triacetoxyborohydride (0.09 mmol) was added at room temperature for 3 hours, L) and saturated sodium bicarbonate (0.2011, 0.766) was reacted for 1 hour at room temperature, sodium triacetoxyborohydride (0.7g, 2.09mmol) was added to the reaction solution, dichloromethane (20 m) was extracted with ethyl bicarbonate (7-2-hydroxy-2-ethyl-2-yl) to obtain a yellow filtrate, the title compound was purified by silica-2-ethyl acetate (7-azaspiro [ 7-2-yl ] column chromatography, 7-2-hydroxy-2, 7-2-hydroxy-2, 7-2-hydroxy-2-ethyl-2-1-hydroxy-1-hydroxy-1-2-1-2-1-2-ethyl-4-one, 7-2-7-silica was added to obtain a yellow silica gel, 7-silica gel, 7-silica gel.
1H NMR(400MHz，CD3OD)7.79(s，1H)，7.31(dd，2H)，7.27(s，1H)，7.09(dd，2H)，6.93(dd，2H)，6.87(d，1H)，6.68(d，1H)，5.11-4.99(m，1H)，4.75(s，1H)，3.84(s，2H)，3.74(s，3H)，2.92-2.77(m，2H)，2.68(t，2H)，2.59(t，2H)，2.45(d，4H)，2.33-2.22(m，2H)，1.81(dd，2H)，1.66(t，2H)，1.59(s，2H).
LCMS m/z＝407.3[(M+2)/2]。
Example 5: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate; bitrifluoroacetate salt (Compound 5)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]meth yl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000401
Figure GPA0000234146600000411
The first step is as follows: 5- (methylamino) pentanoic acid (5B)
5-(methylamino)pentanoic acid
Figure GPA0000234146600000412
1-methyl-2-piperidone (5A) (7.0g, 61.9mmol) was added to an aqueous solution (17m L) of sodium hydroxide (2.7g, 68.0mmol), heated to 110 ℃ for 2 days, cooled to 0 ℃ and adjusted to pH about 2 with concentrated hydrochloric acid, and then directly concentrated under reduced pressure to give the title compound 5- (methylamino) pentanoic acid (5B) as a white solid (8.1g, 100% yield).
LCMS m/z＝132.1[M+1]。
The second step is that: 5- (methylamino) pentanoic acid methyl ester (5C)
methyl 5-(methylamino)pentanoate
Figure GPA0000234146600000413
5- (methylamino) pentanoic acid (5B) (8.0g, 61.0mmol) was dissolved in anhydrous methanol (100m L), cooled to 0 deg.C, sulfoxide chloride (7.3g, 61.0mmol) was added, warmed to 50 deg.C, stirred for 2 hours, concentrated directly under reduced pressure, then concentrated continuously with dichloromethane (150m L× 3) to remove methanol contained in the product to give the title product methyl 5- (methylamino) pentanoate (5C) as an off-white solid (8.8g, 100% yield).
LCMS m/z＝146.2[M+1]。
The third step: 5- [ methyl (prop-2-enoyl) amino ] pentanoic acid methyl ester (5D)
methyl 5-[methyl(prop-2-enoyl)amino]pentanoate
Figure GPA0000234146600000421
5- (methylamino) pentanoic acid methyl ester (5C) (5.9g, 40.0mmol) and acrylic acid (1.44g, 20.0mmol) were dissolved in dichloromethane (25m L), 2- (7-azobisbenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (HATU, CAS: 148893-10-1) (11.4g, 30mmol) was added, cooled to 0 deg.C, N, N-diisopropylethylamine (20.7g, 160mmol) was added dropwise, reaction was allowed to proceed at room temperature for 3 hours, dichloromethane (120m L) and water (100m L) were added to the reaction solution, the fractions were extracted, washed with saturated aqueous sodium chloride solution (100m L× 1), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified on silica gel (eluent ethyl acetate: petroleum ether (v/v): 0: 1-1: 3) to give the title compound 5- [ methyl (propionoyl) amino ] pentanoic acid methyl ester (5- [ 5g, 5.8% yield by column chromatography (1.8%).
1H NMR(400MHz，CDCl3)6.56(m，1H)，6.31(d，1H)，5.67(dd，1H)，3.67(s，3H)，3.49-3.32(m，2H)，3.09-2.96(m，3H)，2.35(t，2H)，1.62(d，4H).
LCMS m/z＝200.1[M+1]。
The fourth step: 5- [ methyl (prop-2-enoyl) amino ] pentanoic acid (5E)
5-[methyl(prop-2-enoyl)amino]pentanoic acid
Figure GPA0000234146600000422
Methyl 5- [ methyl (prop-2-enoyl) amino ] pentanoate (5D) (1.8g, 9.0mmol) was dissolved in tetrahydrofuran (20m L), an aqueous solution of lithium hydroxide (0.32g, 14mmol) was added (40m L) and reacted at room temperature for 3 hours, cooled to 0 ℃ to adjust pH to about 3, dichloromethane (50m L× 3) was added for extraction, the organic layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 5- [ methyl (prop-2-enoyl) amino ] pentanoic acid (5E) as a brown oil (1.48g, 88% yield).
LCMS m/z＝186.1[M+1]。
The fifth step: 5- [3- [2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] non-7-yl ] propionyl-methyl-amino ] pentanoic acid (5F)
5-[3-[2-[2-hydroxy-2，2-bis(2-thienyl)acetyl]oxy-7-azaspiro[3.5]nonan-7-yl]propanoyl-methyl-amino]pentanoic acid
Figure GPA0000234146600000423
5- [ methyl (prop-2-enoyl) amino ] pentanoic acid (5E) (0.76g, 4.13mmol) and 7-azaspiro [3.5] nonan-2-yl 2-hydroxy-2, 2-bis (2-thienyl) acetate (1D) (1.00g, 2.75mmol) were dissolved in 2-methyltetrahydrofuran (20m L), N-diisopropylethylamine (0.57g, 5.5mmol) was added, placed in a microwave reactor, allowed to react at elevated temperature to 100 ℃ for 1 hour, cooled to room temperature, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent dichloromethane/methanol (v: v) ═ 1/0-9/1) to give the title compound 5- [3- [2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] non-7-yl ] propionyl-methyl-amino ] pentanoic acid (5F) as a pale yellow solid (1.1g, 73% yield).
1H NMR(400MHz，CDCl3)7.29(dd，2H)，7.16(dd，2H)，7.02-6.92(m，2H)，5.13(dd，1H)，3.34(m，2H)，3.06(m，2H)，3.00-2.89(m，3H)，2.80(m，6H)，2.47-2.23(m，4H)，1.97-1.71(m，6H)，1.62(m，4H).
LCMS m/z＝601.2[M+1]。
And a sixth step: [7- [3- [ [5- [4- (1, 3-dioxolan-2-yl) aniline ] -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (5H)
[7-[3-[[5-[4-(1，3-dioxolan-2-yl)anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000431
Dissolving 5- [3- [2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] non-7-yl ] propanoyl-methyl-amino ] pentanoic acid (5F) (1.0G, 1.82mmol) and 4- (1, 3-dioxolan-2-yl) aniline (5G) (0.60G, 3.64mmol) in dichloromethane (25m L), cooling to 0 ℃, adding 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (HATU, CAS: 148893-10-1) (1.04G, 2.73mmol), dropwise adding N, N-diisopropylethylamine (0.47G, 3.65mmol), reacting at room temperature for 3 hours, adding dichloromethane (120m L) and water (100m L) to the reaction solution, extracting the fractions, the organic phase again with saturated aqueous sodium chloride solution (100m L×), washing the filtrate without water, filtering, purifying the residue by silica-2-oxo-2-pentyl-aniline (3H) -2-yl) column chromatography to obtain the title compound as yellow solid (3G, 3.5-oxo-2-pentyl-nonane-2-yl).
1H NMR(400MHz，CDCl3)7.62(d，2H)，7.41(t，2H)，7.29(m，2H)，7.17(m，2H)，6.98(dd，2H)，5.76(s，1H)，5.12(m，1H)，4.10(m，2H)，4.02(m，2H)，3.39(m，2H)，2.97(s，3H)，2.68(m，2H)，2.54(m，2H)，2.36(m，8H)，1.85(m，2H)，1.71(m，4H)，1.57(m，4H).
LCMS m/z＝696.3[M+1]。
The seventh step: [7- [3- [ [5- (4-carboxanilide) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (5I)
[7-[3-[[5-(4-formylanilino)-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000441
[7- [3- [ [5- [4- (1, 3-dioxolan-2-yl) aniline ] -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (5H) (1.1g, 1.6mmol) was dissolved in acetonitrile (40M L), a 3M aqueous hydrochloric acid solution (20M L) was added dropwise, reaction was carried out at room temperature for 1 hour, dichloromethane (100M L) and water (100M L) were added to the reaction solution, the layers were extracted, the organic phase was washed with a saturated aqueous sodium chloride solution (100M L× 1) and dried over anhydrous sodium sulfate, filtration was carried out, and the filtrate was concentrated under reduced pressure to give the title compound [7- [3- [ [5- (4-formanilide) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2-acetate (2-thienyl) as a yellow solid (95% yield).
1H NMR(400MHz，CDCl3)9.89(s，1H)，7.94(dd，2H)，7.81(dd，2H)，7.30(m，2H)，7.15(m，2H)，6.98(m，2H)，5.13(m，1H)，3.29(m，4H)，3.09(t，2H)，2.98(m，3H)，2.59(m，4H)，2.32(m，4H)，1.98(m，2H)，1.75(m，8H).
LCMS m/z＝652.2[M+1]。
Eighth step: [7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] anilino ] -5-oxo-pentyl ] -methyl-amino ] -3 oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (5K)
[7-[3-[[5-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000442
[7- [3- [ [5- (4-carboxanilide) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (5I) (0.25G, 0.38mmol) and 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.19G, 0.58mmol) were dissolved in a mixed solvent of isopropanol/dichloromethane (v/v ═ 1: 1, 10m L), stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (0.24G, 1.14mmol) was added, stirred for 2 hours dichloromethane (150m L) and water (50m L) were added to the reaction solution, the organic phase was extracted with saturated sodium chloride aqueous solution (50m L× G) and the filtrate was washed with anhydrous sodium sulfate, filtered, and the residue was purified by silica-based chromatography to obtain the title compound [7- [ 3-oxo-propyl ] -7-azaspiro [ 3-amino-2-ethyl ] -7-amino-1- [ tert-butyl (dimethyl) ethyl- [ [ -1- [ 7-amino-2-yl ] quinoline (v- [ [).
The ninth step: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate; bitrifluoroacetate salt (Compound 5)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000451
[7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] anilino ] -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (5K) (0.17g, 0.18mmol) was dissolved in dichloromethane (10m L), triethylamine trihydrofluoride salt (0.14g, 0.88mmol) was added, reaction was allowed to proceed overnight at room temperature, the reaction solution was adjusted with saturated sodium bicarbonate, extracted with 8% methanol/dichloromethane (v/v ═ 8: 92, 100m L), the organic phase was washed with saturated aqueous sodium chloride solution (50m L×), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was isolated with a liquid preparative column (preparative run-preparative C: 36% aqueous phase eluent-2-ethyl-2-oxo-2-5-quinolinyl-2- (8-hydroxy-2-oxo-5-yl) ethyl ] 2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (0.05% aqueous eluent-2.
1H NMR(400MHz，CD3OD)8.06(dd，1H)，7.54(dd，2H)，7.32(dd，2H)，7.25(dd，2H)，7.13(d，1H)，7.01(m，2H)，6.87(m，3H)，6.49(dd，1H)，5.25(m，1H)，4.99(dt，1H)，4.13(d，2H)，3.30(m，6H)，3.07(d，2H)，2.82(m，7H)，2.32(m，3H)，2.21(s，1H)，1.82(dd，3H)，1.70(m，3H)，1.55(m，4H)..
LCMS m/z＝428.9[(M+2)/2]。
Example 6: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate; bitrifluoroacetate salt (Compound 6)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000461
Compound 6
Figure GPA0000234146600000462
The first step is as follows: [7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (6B)
[7-[3-[[5-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000471
[7- [3- [ [5- (4-carboxanilide) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (5I) (0.2g, 0.3mmol) and 8- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -5-hydroxy-4H-1, 4-benzoxazin-3-one (6A) (see WO2008149110a intermediate 65) were prepared (0.2g, 0.5mmol) dissolved in a mixed solvent of isopropanol/dichloromethane (v/v ═ 1: 1, 10m L), stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (0.19g, 0.9mmol) was added to the reaction solution and dichloromethane (150m L) and water (50m ═ L) was added to extract the organic phase with sodium chloro-2- [3- [ [ -2-oxo-propyl ] -7-azaspiro [ 3-yl ] 2-yl ] acetate (0.2-7-hydroxy-2, 0.7-7-4H-phenyl ] benzoxazin 2 intermediate 65) yield, then sodium triethyloxyborohydride was added to the reaction solution, and the reaction solution was concentrated under reduced pressure to obtain a saturated sodium hydroxide solution (7- [3- [ [ -4-7- [ 7-4-methyl-7-4-ethyl ] nona-2-7-4-phenyl ] nona-7-4-2-4-7-ethyl ] solid.
LCMS m/z＝487.9[(M+2)/2]。
The second step is that: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate; bitrifluoroacetate salt (Compound 6)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000472
[7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (6B) (0.2g, 0.2mmol) was dissolved in tetrahydrofuran (5m L), triethylamine trihydrofluoride salt (0.2g, 1.0mmol) was added, the reaction was allowed to react overnight at room temperature, the reaction solution was adjusted with saturated sodium bicarbonate, filtration was performed, the resulting solid was dissolved with 8% methanol/dichloromethane (v/v ═ 8: 92, 50m L), saturated aqueous sodium chloride solution (50m L×) was washed with anhydrous sodium sulfate, the filtrate was filtered, the filtrate was concentrated under reduced pressure to prepare a liquid phase containing the compound under a gradient of 7-hydroxy-2-ethyl-2-oxo-2-1, 4H-1, 4-benzoxazin a-bis (2-hydroxy-2-yl) -2-hydroxy-2-yl) acetate (0.2-hydroxy-2-yl) was dissolved in tetrahydrofuran (0.5% TFA) and the title compound was eluted under a gradient to obtain a (5-7-5-hydroxy-7-hydroxy-7-hydroxy-7% aqueous phase).
1H NMR(400MHz，CD3OD)7.68(dd，2H)，7.45(dd，2H)，7.40(dd，2H)，7.16(t，2H)，7.01(m，3H)，6.58(d，1H)，5.16(m，2H)，4.49(m，2H)，4.23(m，2H)，3.44(m，6H)，3.17(dd，1H)，2.98(m，8H)，2.47(m，3H)，2.37(m，1H)，1.98(m，3H)，1.84(d，3H)，1.70(m，4H).
LCMS m/z＝430.8[(M+2)/2]。
Free base of compound 6: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000481
Dissolving [7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (6B) (40.0g, 41.1mmol) in tetrahydrofuran (500m L), adding triethylamine trihydrofluoride (33.1g, 205mmol), reacting overnight at room temperature, adjusting the reaction solution with saturated sodium bicarbonate, filtering, preparing a solid with 8% methanol/dichloromethane (v/v ═ 8: 92, 50m L), washing with saturated aqueous sodium chloride solution (250m L× 1), drying with anhydrous sodium sulfate, filtering, preparing a filtrate under reduced pressure, preparing a residual liquid phase with 8% methanol/dichloromethane (v ═ 8: 92: 2: L), purifying the obtained solid phase with dichloromethane (3-ethyl-2-hydroxy-2-oxo-2-1H-1, 4 ═ 2-oxa-2-yl) acetate (40% of ethyl-2-ethyl-2-hydroxy-2-yl) after removing the obtained solid phase, dissolving the obtained solid phase, removing the obtained after the title compound (3-ethyl-hydroxy-2-hydroxy-2-hydroxy-2-yl) in tetrahydrofuran (3-hydroxy-2-yl) is dissolved in tetrahydrofuran (3-hydroxy-2-hydroxy-2-yl) in tetrahydrofuran (35% TFA gradient, removing the anhydrous sodium chloride, the obtained, the title compound, the anhydrous sodium bicarbonate solution, the obtained after the procedure of ethyl-2-hydroxy-2-hydroxy-2-.
1H NMR(400MHz，CD3OD)7.44(d，1H)，7.42(d，1H)，7.26(m，2H)，7.18(s，1H)，7.16(s，1H)，7.03(m，2H)，6.89-6.85(m，2H)，6.82(d，1H)，6.41(d，1H)，5.01-4.91(m，2H)，4.39-4.35(m，2H)，3.74-3.63(m，2H)，3.31(m，2H)，3.25(s，1H)，2.94(s，2H)，2.73-2.59(m，2H)，2.58-2.49(m，2H)，2.49-2.43(m，2H)，2.37-2.11(m，8H)，1.70(m，2H)，1.51(m，8H).
LCMS m/z＝430.9[(M+2)/2]。
Example 7: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (4-hydroxy-2-oxo-3H-1, 3-benzothiazol-7-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] -2-hydroxy-2, 2-di (2-thienyl) acetate; bitrifluoroacetate salt (Compound 7)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(4-hydroxy-2-oxo-3H-1，3-benzothiazol-7-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroacetic acid
Figure GPA0000234146600000491
Compound 7
Figure GPA0000234146600000492
Figure GPA0000234146600000501
The first step is as follows: 7- [ (1R) -2-azido-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -4- [ tert-butyl (dimethyl) silyl ] oxy-3H-1, 3-benzothiazol-2-one (7B)
7-[(1R)-2-azido-1-[tert-butyl(dimethyl)silyl]oxy-ethyl]-4-[tert-butyl(dimethyl)silyl]oxy-3H-1.3-benzothiazol-2-one
Figure GPA0000234146600000502
7- [ (1R) -2-azido-1-hydroxy-ethyl ] -4-hydroxy-3H-1, 3-benzothiazol-2-one (7A) (prepared by reference to WO2009098448A 1) (0.56g.2.2mmol) was dissolved in N, N-dimethylformamide (20m L), then imidazole (0.6g, 8.9mmol) was added, tert-butyldimethylchlorosilane (1.3g, 8.9mmol) was added in portions, a catalytic amount of 4-dimethylaminopyridine was added, the temperature was raised to 40 ℃ and stirred for 7 hours, the reaction solution was poured into water (100m L), extracted with ethyl acetate (100m L× 1), the organic phase was washed with a saturated aqueous sodium chloride solution (100m L× 2), dried with anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was subjected to silica gel column chromatography (eluent ethyl acetate/petroleum ether (v/v) ═ 0/1 to 5/95), to give the title compound 7- [ (1R) -2-azido-1- [ tert-butyloxy ] dimethyl-ethyl ] benzothiazol-2-one (silica-based solid yield 80%).
1H NMR(400MHz，CDCl3)8.25(s，1H)，6.92(d，1H)，6.71(d，1H)，4.78(dd，1H)，3.41(dd，1H)，3.25(dd，1H)，1.05-0.98(m，9H)，0.92-0.88(m，9H)，0.28(t，6H)，0.12(d，3H)，-0.04(d，3H).
The second step is that: 7- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -4- [ tert-butyl (dimethyl) silyl ] oxy-3H-1, 3-benzothiazol-2-one (7C)
7-[(1R)-2-amino-1-[tert-butyl(dimethyl)silyl]oxy-ethyl]-4-[tert-butyl(dimethyl)silyl]oxy-3H-1，3-benzothiazol-2-one
Figure GPA0000234146600000511
7- [ (1R) -2-azido-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -4- [ tert-butyl (dimethyl) silyl ] oxy-3H-1, 3-benzothiazol-2-one (7B) (0.85g, 1.8mmol) was dissolved in ethyl acetate (20m L), 10% (w/w) palladium on carbon (0.085g) was added and stirred under a balloon of hydrogen at normal pressure overnight, filtered through celite, and concentrated to give the title compound 7- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -4- [ tert-butyl (dimethyl) silyl ] oxy-3H-1, 3-benzothiazol-2-one (7C) as a light black solid (0.7g, 90% yield).
1H NMR(400MHz，CDCl3)6.89(d，1H)，6.68(t，1H)，4.64(dd，1H)，2.88(ddd，2H)，1.04-0.96(m，9H)，0.95-0.87(m，9H)，0.33-0.23(m，6H)，0.12-0.06(m，3H)，-0.04--0.11(m，3H).
The third step: [7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- [4- [ tert-butyl (dimethyl) silyl ] oxy-2-oxo-3H-1, 3-benzothiazol-7-yl ] ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (7D)
[7-[3-[[5-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-[4-[tert-butyl(dimethyl)silyl]oxy-2-oxo-3H-1，3-benzothiazol-7-yl]ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000512
[7- [3- [ [5- (4-carboxanilide) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (5I) (0.25g, 0.38mmol) and 7- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -4- [ tert-butyl (dimethyl) silyl ] oxy-3H-1, 3-benzothiazol-2-one (7C) (0.26g, 0.58mmol) were dissolved in a mixed solvent of isopropanol/dichloromethane (v/v 1: 110m L), stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (0.24g, 1.14mmol) was added to the reaction solution, dichloromethane (150m L) and water (50m L) was added to the reaction solution, the aqueous phase was separated by filtration, and the title compound was concentrated under reduced pressure to give a title compound [3- [ [ -oxo-propyl ] -7-amino-2-yl ] 2-phenyl ] -7- [ (1R) -2-tert-butyl (dimethyl) amino-2-silyl ] oxy-2-ethyl ] -7- [ (3-2-yl ] phenyl ] 2-oxo-7-amino-2-yl ] phenyl ] ethyl ] -7- [ (1- [ 2-tert-butyl (v/v ] [ (1: 110m L) as a solid.
LCMS m/z＝488.8[(M+2)/2]。
The fourth step: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (4-hydroxy-2-oxo-3H-1, 3-benzothiazol-7-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate; bitrifluoroacetate salt (Compound 7)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(4-hydroxy-2-oxo-3H-1，3-benzothiazol-7-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroacetic acid
Figure GPA0000234146600000521
[7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- [4- [ tert-butyl (dimethyl) silyl ] oxy-2-oxo-3H-1, 3-benzothiazol-7-yl ] ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (7D) (0.35g, 0.32mmol) was dissolved in tetrahydrofuran (5m L), triethylamine trihydrofluoride salt (0.26g, 1.6mmol) was added, the reaction was allowed to react overnight at room temperature, the reaction solution was adjusted with a saturated sodium bicarbonate solution, the organic phase was extracted with a saturated aqueous sodium chloride solution (50m L×) and washed with anhydrous sodium sulfate, the filtrate was filtered, the filtrate was prepared with 8% methanol/dichloromethane (v/v ═ 8: 92, 100m L) and the resulting filtrate was purified by column chromatography using a gradient elution under reduced pressure to obtain a white liquid phase containing 3- [4- [ [ -ethyl ] -2-oxo-2-oxo-propyl ] -7H-hydroxy-2, and a-2-hydroxy-2, after elution under reduced pressure (0.5% elution conditions of 0.5% concentration of ethyl-7% TFA) to obtain a, 2-7% TFA, 2-7% aqueous phase, the title compound (7% aqueous phase).
1H NMR(400MHz，CD3OD)7.69(dd，2H)，7.46(dd，2H)，7.40(dd，2H)，7.16(t，2H)，6.99(dd，3H)，6.77(d，1H)，5.14(m，1H)，4.99(dd，1H)，4.25(d，2H)，3.44(m，6H)，3.13(m，2H)，3.03(m，3H)，2.93(m，4H)，2.48(m，3H)，2.36(m，1H)，1.98(m，3H)，1.84(m，3H)，1.70(m，4H).
LCMS m/z＝431.8[(M+2)/2]。
Example 8: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] aniline ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate; bitrifluoroacetate salt (Compound 8)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000531
Compound 8
Figure GPA0000234146600000532
The first step is as follows: 3- [2- [ (2-Phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propionic acid (8A)
3-[2-[(2-phenylphenyl)carbamoyloxy]-7-azaspiro[3.5]nonan-7-yl]propanoic acid
Figure GPA0000234146600000541
7-Azaspiro [3, 5] nonan-2-yl N- (2-phenylphenyl) carbamate (2D) (0.8g, 2mmol) and acrylic acid (2.0g, 20mmol) were dissolved in 2-methyltetrahydrofuran (10m L), placed in a microwave reactor, heated to 100 ℃ for reaction for 1 hour, cooled to room temperature, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (methanol: dichloromethane (v/v) ═ 0: 1 to 1: 9) to give the title compound 3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propionic acid (8A) as a pale yellow solid (0.8g, 80% yield).
1H NMR(400MHz，CDCl3)8.08(d，1H)，7.50(m，2H)，7.42(m，1H)，7.35(m，3H)，7.21(dd，1H)，7.13(m，1H)，6.63(s，1H)，4.99(m，1H)，2.96(m，8H)，2.37(m，2H)，1.88(m，6H).
LCMS m/z＝409.1[M+1]。
The second step is that: 5- [ methyl- [3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propionyl ] amino ] pentanoic acid methyl ester (8B)
methyl 5-[methyl-[3-[2-[(2-phenylphenyl)carbamoyloxy]-7-azaspiro[3.5]nonan-7-yl]propanoyl]amino]pentanoate
Figure GPA0000234146600000542
3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propionic acid (8A) (0.8g, 2mmol) and methyl 5- (methylamino) valerate (0.6g, 4mmol) were dissolved in dichloromethane (25m L), 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (1g, 3mmol) was added, cooled to 0 ℃, diisopropylethylamine (2.0g, 20mmol) was added dropwise, reaction was allowed to warm to room temperature for 3 hours, dichloromethane (120m L) and water (100m L) were added to the reaction solution, the layers were extracted, the organic phase was washed with a saturated aqueous sodium chloride solution (100m L× 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 5- [ methyl- [3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propionyl ] amino ] pentanoic acid methyl ester (8 g, yield 0.100%).
LCMS m/z＝536.2[M+1]。
The third step: 5- [ methyl- [3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propionyl ] amino ] pentanoic acid (8C)
5-[methyl-[3-[2-[(2-phenylphenyl)carbamoyloxy]-7-azaspiro[3.5]nonan-7-yl]propanoyl]amino]pentanoic acid
Figure GPA0000234146600000551
Methyl 5- [ methyl- [3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propanoyl ] amino ] pentanoate (8B) (1.0g, 1.86mmol) was dissolved in tetrahydrofuran (10m L), an aqueous solution (10m L) of sodium hydroxide (0.37g, 9.3mmol) was added, the reaction was carried out at room temperature for 2 hours, cooling was carried out to 0 ℃ to adjust the pH to about 3, dichloromethane (50m L× 3) was added for extraction, the organic layers were combined, dried over anhydrous sodium sulfate, filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent dichloromethane/methanol (v: v) 1/0 to 9/1) to give the title compound 5- [ methyl- [3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propanoyl ] amino ] pentanoic acid (8C) as a pale yellow solid (0.85g, yield 87%).
1H NMR(400MHz，CDCl3)8.03(s，1H)，7.50(dd，2H)，7.42(t，1H)，7.35(m，3H)，7.21(dd，1H)，7.13(t，1H)，6.65(d，1H)，4.99(m，1H)，3.47(m，2H)，3.37(s，2H)，3.30(m，2H)，3.00(m，4H)，2.89(d，3H)，2.83(m，2H)，2.39(m，4H)，2.00(m，2H)，1.87(m，2H)，1.81(m，2H)，1.66(s，2H).
LCMS m/z＝522.2[M+1]。
The fourth step: [7- [3- [ [5- [4- (1, 3-dioxolan-2-yl) anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8D)
[7-[3-[[5-[4-(1，3-dioxolan-2-yl)anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000552
5- [ methyl- [3- [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] propanoyl ] amino ] pentanoic acid (8C) (0.4g, 0.8mmol) and 4- (1, 3-dioxolan-2-yl) aniline (0.3g, 1.6mmol) were dissolved in dichloromethane (10m L), cooled to 0 deg.C, 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (0.44g, 1.0mmol) was added dropwise N, N-diisopropylethylamine (0.2g, 2mmol), reacted at room temperature for 3 hours, dichloromethane (50m L) and water (50m L) were added to the reaction solution, the layers were extracted, the organic layer was washed with a saturated aqueous sodium chloride solution (50m L×), dried without water, filtered, the filtrate was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (eluent dichloromethane/methanol (v/v) ═ 7-azaspiro [3.5] phenyl ] -7-yl ] nonanoate (3-7-phenyl-7-yl) as a yellow solid (3 g, 7-azaspiro [ 7-yl ] phenyl-7-phenyl-7-yl ] nonanoate (yield).
1H NMR(400MHz，CDCl3)8.10(d，1H)，7.63(dd，2H)，7.50(dd，2H)，7.38(m，6H)，7.21(dd，1H)，7.13(m，1H)，6.61(d，1H)，5.75(d，1H)，4.97(m，1H)，4.10(m，2H)，4.00(m，2H)，3.46(m，1H)，3.31(m，1H)，3.00(s，3H)，2.77(m，2H)，2.72(m，2H)，2.58(m，2H)，2.48(m，2H)，2.29(m，4H)，1.78(m，4H)，1.63(m，6H).
LCMS m/z＝669.4[M+1]。
The fifth step: [7- [3- [ [5- (4-formylanilino) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8E)
[7-[3-[[5-(4-formylanilino)-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000561
[7- [3- [ [5- [4- (1, 3-dioxolan-2-yl) anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8D) (1.2g, 1.8mmol) was dissolved in acetonitrile (20M L), a 3M aqueous hydrochloric acid solution (6M L) was added dropwise, reaction was carried out at room temperature for 1 hour, methylene chloride (100M L) and water (100M L) were added to the reaction solution, the layers were separated by extraction, the organic phase was washed with a saturated aqueous sodium chloride solution (100M L× 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound [7- [3- [ [5- (4-formylanilino) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenyl) carbamate (8E) in a yellow solid (yield (89 g, 0.89%).
LCMS m/z＝625.3[M+1]。
And a sixth step: [7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8F)
[7-[3-[[5-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000571
[7- [3- [ [5- (4-formylanilino) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8E) (0.25G, 0.40mmol) and 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.20G, 0.60mmol) were dissolved in a mixed solvent of isopropanol/dichloromethane (v/v ═ 1: 1, 10 m- [ [ 36), stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (0.25G, 1.2mmol) was added, further stirred for 2 hours, dichloromethane (150m L) and water (50m L) were added to the reaction liquid, the layers were extracted, the organic phase was washed with a saturated aqueous sodium chloride solution (50m L× G), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure to obtain a crude product of dichloromethane (7G-phenyl-ethyl-7- [ 3-phenyl ] -7-azaspiro [ 3-yl ] N- (2-phenyl) carbamate (0.20G, 7-ethyl-7-phenyl) as a solid, and the title compound was purified.
LCMS m/z＝472.4[(M+2)/2]。
The seventh step: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] aniline ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate; bitrifluoroacetate salt (Compound 8)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000581
[7- [3- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8F) (0.11g, 0.12mmol) was dissolved in dichloromethane (10m L), triethylamine trihydrofluoride salt (0.10g, 0.58mmol) was added, TFA was reacted overnight at room temperature, the reaction solution was adjusted with saturated sodium bicarbonate solution, extracted with 8% methanol/dichloromethane (v/v ═ 8: 92, 100m L), the organic phase was washed with saturated chlorinated solution (50m 32 1), anhydrous sodium sulfate was dried, filtered, the filtrate was concentrated under reduced pressure, the residue was separated with a liquid preparative column (reverse phase preparative conditions: C: 36C, aqueous phase preparative column chromatography, aqueous eluent-ethyl-2-amino-2-ethyl-2-8-phenyl-yl ] N- (2-phenyl) carbamate (0.11-8% ethyl-phenyl) ethyl-8% ethyl-8-phenyl) carbamate, 0.5-8% aqueous eluent (0.5% aqueous eluent-ethyl-2-ethyl-7-phenyl) was obtained, the title compound was purified.
1H NMR(400MHz，CD3OD)8.20(t，1H)，7.69(dd，2H)，7.57(s，1H)，7.38(m，11H)，7.04(d，1H)，6.63(dd，1H)，5.38(m，1H)，4.86(s，1H)，4.26(d，2H)，3.43(m，6H)，3.20(d，2H)，3.07(s，2H)，3.01(d，1H)，2.97(m，2H)，2.92(m，2H)，2.46(dd，3H)，2.23(s，1H)，1.91(m，6H)，1.70(d，4H).
LCMS m/z＝415.3[(M+2)/2]。
Example 9: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate; bitrifluoroacetate salt (Compound 9)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000591
Compound 9
Figure GPA0000234146600000592
The first step is as follows: [7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (9A)
[7-[3-[[5-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000593
[7- [3- [ [5- (4-formylanilino) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8E) (0.25g, 0.3mmol) and 8- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -5-hydroxy-4H-1, 4-benzoxazin-3-one (6A) (0.2g, 0.6mmol) were dissolved in a mixed solvent of methanol/dichloromethane (v/v ═ 1/1, 10m L), stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (0.25g, 1.2mmol) was added, further stirred for 3 hours dichloromethane (150m L) and water (50m L) were added to the reaction liquid, the layers were extracted, the organic phase was washed with a saturated sodium chloride aqueous solution (50m L×), dried without water, the filtrate, and the title compound was purified by column chromatography (3- [ [ -ethyl-phenyl) -3-oxo-propyl ] -7-azaspiro [ 3-amino ] -2-propyl ] -7-azaspiro [ 3-amino-1- [ tert-butyl (2-yl ] carba-2-silica-yl ] N- (2-yl ] amino-3-one (0.2-ethyl-2-silica-2-silica-2-one) (0.2-2-ethyl) yield).
LCMS m/z＝474.3[(M+2)/2]。
The second step is that: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate; bitrifluoroacetate salt (Compound 9)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000601
[7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (9A) (0.11g, 0.12mmol) was dissolved in tetrahydrofuran (5m L), triethylamine trihydrofluoride salt (0.1g, 0.58mmol) was added and reacted overnight at room temperature, the reaction solution was adjusted with a saturated sodium bicarbonate solution, the filtrate was filtered, the resulting solid was dissolved with 8% methanol/dichloromethane (v/v ═ 8: 92, 50m L), a saturated sodium chloride aqueous solution (50m L×) was washed, the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was prepared by column chromatography (v/v ═ 8 ═ v ═ 92, 50m L) and the title compound was purified by column chromatography (3-ethyl-phenyl-ethyl-2-amino-2-phenyl) under the conditions of title product was obtained as a white-hydroxy-phenyl-2-oxaspiro [ 3-2-ethyl ] -7-azaspiro [ 3-5-oxanonan- [ 3-5-phenyl ] carbamate (0.5-5-7-phenyl) solid, TFA-7-phenyl) and the title compound was dissolved in a-7% yield, and the title compound was purified by the title product was obtained as a-7% eluent under the title.
1H NMR(400MHz，CD3OD)7.69(dd，2H)，7.58(s，1H)，7.37(m，10H)，7.02(d，1H)，6.58(d，1H)，5.17(d，1H)，4.86(s，1H)，4.49(m，2H)，4.23(m，2H)，3.43(m，6H)，3.16(dd，1H)，2.98(m，8H)，2.46(m，3H)，2.23(s，1H)，1.91(m，6H)，1.70(d，4H).
LCMS m/z＝417.3[(M+2)/2]。
Example 10: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (4-hydroxy-2-oxo-3H-1, 3-benzothiazol-7-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate; bitrifluoroacetate salt (Compound 10)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(4-hydroxy-2-oxo-3H-1，3-benzothiazol-7-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000611
Compound 10
Figure GPA0000234146600000612
The first step is as follows: [7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2 [4- [ tert-butyl (dimethyl) silyl ] oxy-2-oxo-3H-1, 3-benzothiazol-7-yl ] ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (10A)
[7-[3-[[5-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-[4-[tert-butyl(dimethyl)silyl]oxy-2-oxo-3H-1，3-benzothiazol-7-yl]ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000613
[7- [3- [ [5- (4-formylanilino) -5-oxo-pentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (8E) (0.25g, 0.38mmol) and 7- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxyethyl ] -4- [ tert-butyl (dimethyl) silyl ] oxy-3H-1, 3-benzothiazol-2-one (7C) (0.27g, 0.6mmol) were dissolved in a mixed solvent of methanol/dichloromethane (v/v ═ 1/1, 10m L), stirred at room temperature for 1 hour, then sodium triacetoxyborohydride (0.25g, 1.2mmol) was added, reaction was carried out at room temperature for 2 hours, dichloromethane (150m L) and water (50m L) were added to the reaction solution, the layers were extracted, and the organic phase was washed with a saturated aqueous sodium chloride solution (50m 2) to obtain a filtrate, and the title compound was concentrated under reduced pressure to give a 3- [ [ -phenyl ] amino ] -3-propyl ] -7-azaspiro [ 3-amino-2-yl ] N- (2-yl ] carbamate (0.7-phenyl) solid (v- [ [ -2-propyl ] -4-phenyl-2-phenyl-oxo-2-phenyl).
The second step is that: [7- [3- [ [5- [4- [ [ [ (2R) -2-hydroxy-2- (4-hydroxy-2-oxo-3H-1, 3-benzothiazol-7-yl) ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate; bitrifluoroacetate salt (Compound 10)
[7-[3-[[5-[4-[[[(2R)-2-hydroxy-2-(4-hydroxy-2-oxo-3H-1，3-benzothiazol-7-yl)ethyl]amino]methyl]anilino]-5-oxo-pentyl]-methyl-amino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenvlphenvl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000621
[7- [3- [ [5- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2 [4- [ tert-butyl (dimethyl) silyl ] oxy-2-oxo-3H-1, 3-benzothiazol-7-yl ] ethyl ] amino ] methyl ] anilino ] -5-oxopentyl ] -methyl-amino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenylphenyl) carbamate (10A) (0.4g, 0.4mmol) was dissolved in dichloromethane (10m L), triethylamine trihydrofluoride salt (TFA 0.3g, 2mmol) was added and reacted overnight at room temperature, the reaction solution was adjusted to base with a saturated sodium bicarbonate solution, the reaction solution was extracted with 8% methanol/dichloromethane (v/v ═ dichloromethane (8: 92, 100m L), the organic phase was washed with a saturated sodium chloride aqueous solution (50m L×) and dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the residue was isolated with a preparative column (v/v ═ 8: 92, 100m L min, the eluent) aqueous phase was eluted as a white solid-containing 3-hydroxy-ethyl-2-phenyl-2-ethyl-2-phenyl-ethyl-2-amino-phenyl-2-phenyl-7-phenyl-7H-amino-phenyl-7-amino-phenyl-7 (TFA) and the title compound was purified at a gradient under the flow rate of 0.5-7% yield 0.5-7-azaspiro [ TFA).
1H NMR(400MHz，CD3OD)7.55(dd，2H)，7.43(s，1H)，7.24(m，9H)，6.83(d，1H)，6.63(d，1H)，4.84(d，1H)，4.72(s，1H)，4.10(s，2H)，3.29(m，6H)，2.93(s，4H)，2.82(m，5H)，2.32(m，3H)，2.09(s，1H)，1.76(m，6H)，1.56(m，4H).
LCMS m/z＝418.3[(M+2)/2]。
Example 11: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (5-hydroxy-3-oxo-4H-1, 4-benzoxazin-8-yl) ethyl ] amino ] methyl ] -5-methoxyanilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] -2-hydroxy-2, 2-di (2-thienyl) acetate (Compound 11)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(5-hydroxy-3-oxo-4H-1，4-benzoxazin-8-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000631
[7- [3- (2-chloro-4-formyl-5-methoxyanilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] -2-hydroxy-2, 2-di (2-thienyl) acetate (1F) (0.300g, 0.497mmol) was dissolved in a mixed solvent of dichloromethane (5m L) and isopropanol (5m L), 8- [ (1R) -2-amino-1-hydroxy-ethyl ] -5-hydroxy-4H-1, 4-benzoxazin-3-one (11A, see WO2009098448a 1) was added (0.134g, 0.597mmol), reaction was stirred at 30 ℃ for 1.5 hours and sodium triacetoxyborohydride (0.316g, 1.49mmol) was added, reaction was continued for 2 hours at 30 ℃, saturated aqueous ammonium chloride solution (15m L) was added dropwise, extraction was performed with dichloromethane (30m 463), organic layers were combined, saturated aqueous sodium oxyborohydride (10g, 1.49mmol) was added, the reaction was washed with saturated aqueous ammonium chloride solution (10g, and the title compound was purified by silica-2-ethyl-7-azaspiro [ 3-2-yl ] ethyl [ (7 ] ethyl-2-hydroxy-7-2-oxo-2-ethyl ] -7-hydroxy-2-oxo-oxadiazin-3-one (11A) as a solid after yield was purified by chromatography, silica gel chromatography (7-silica gel).
1H NMR(400MHz，CD3OD)7.69(s，1H)，7.28-7.25(m，2H)，7.18(s，1H)，7.04-7.01(m，2H)，6.88-6.86(m，2H)，6.79(d，1H)，6.41(d，1H)，5.38(s，1H)，5.05--4.96(m，1H)，4.92-4.86(m，1H)，4.37(s，2H)，3.68(s，3H)，3.66--3.58(m，2H)，3.22-3.18(m，6H)，2.72-2.62(m，2H)，2.62--2.55(m，2H)，2.54-2.48(m，2H)，2.42-2.30(m，3H)，2.28-2.18(m，2H)，1.77-1.73(m，2H)，1.62-1.56(m，2H)，1.54-1.50(m，2H).
LCMS m/z＝406.1[(M+2)/2]。
Example 12: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxyanilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] -2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate; bitrifluoroacetate salt (Compound 12)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentvl-2-hydroxy-2-(2-thienyl)acetate ditrifluoroacetate
Figure GPA0000234146600000641
The first step is as follows: 2-oxo-2- (2-thienyl) acetic acid (12B)
2-oxo-2-(2-thienyl)acetic acid
Figure GPA0000234146600000651
Ethyl 2-oxo-2- (2-thienyl) acetate (6.0g, 32.6mmol) was dissolved in water (20m L), and an aqueous solution of sodium hydroxide (2.61g, 65.1mmol) was added dropwise at room temperature to react for 2 hours after completion of the reaction, water (50m L) was added, the layers were extracted with dichloromethane (50m L× 2), the aqueous phase was adjusted to pH 3 with dilute hydrochloric acid, extracted with dichloromethane (100m L× 2), the second organic phase was combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound, 2-oxo-2- (2-thienyl) acetic acid (12B), a white solid (4.00g, 78.6% yield).
1H NMR(400MHz，DMSO-d6)14.51(s，1H)，8.22(m，1H)，8.07(m，1H)，7.32(m，1H).
LCMS m/z＝157.1[M+1].
The second step is that: 2- [ 2-oxo-2- (2-thienyl) acetoxy ] -7-azaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (12C)
tert-butyl 2-[2-oxo-2-(2-thienyl)acetyl]oxy-7-azaspiro[3.5]nonane-7-carboxylate
Figure GPA0000234146600000652
Dissolving 2-oxo-2- (2-thienyl) acetic acid (12B) (0.500g, 3.20mmol) in dichloromethane (10m L), adding oxalyl chloride (0.813g, 6.40mmol) dropwise, then adding one drop of N, N-dimethylformamide, reacting at room temperature for 1 hour, removing the solvent and excess oxalyl chloride under reduced pressure to give reaction solution 2. dissolving tert-butyl 2-hydroxy-7-azaspiro [3.5] nonane-7-carboxylate (0.773g, 3.20mmol) in dichloromethane (10m L), adding reaction solution 2 under ice bath, reacting at room temperature for 1 hour after completion of addition, adding water (50m L), extracting with dichloromethane (100m L× 2), combining the organic phases, drying over anhydrous sodium sulfate, filtering, concentrating under reduced pressure, separating and purifying the residue by silica gel column chromatography (eluent ethyl acetate: petroleum ether (v/v) ═ 49: 1-10: 1) to give the title compound 2- [ 2-oxo-2-oxospiro [ 2-carboxylic acid- (2-oxospiro [ 3.7 ] nonane ] (yield: 3.7 ] pale yellow).
1H NMR(400MHz，CDCl3)8.12(d，1H)，7.82(d，1H)，7.23-7.17(m，1H)，5.26(m，1H)，3.43-3.35(m，2H)，3.35-3.26(m，2H)，2.52-2.40(m，2H)，2.09-2.01(m，2H)，1.59(m，4H)，1.45(s，9H).
LCMS m/z＝402.1[M+23].
The third step: 2- [ 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetoxy ] -7-azaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (12D)
tert-butyl 2-[2-cyclopentyl-2-hydroxy-2-(2-thienyl)acetyl]oxy-7-azaspiro[3.5]nonane-7-carboxylate
Figure GPA0000234146600000661
Tert-butyl 2- [ 2-oxo-2- (2-thienyl) acetoxy ] -7-azaspiro [3.5] nonane-7-carboxylate (12C) (1.0g, 2.64mmol) was dissolved in tetrahydrofuran (20M L), cooled to-40 ℃ under nitrogen protection, 1.0M tetrahydrofuran solution of cyclopentyl magnesium bromide (3.95M L, 3.95mmol) was added dropwise, after completion of the addition, the temperature was raised to room temperature for reaction for 30 minutes, after completion of the reaction, saturated aqueous ammonium chloride solution (50M L) was added, extraction was performed with ethyl acetate (100M L× 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 (eluent ethyl acetate: petroleum ether (v/v) ═ 49: 1 to 19: 1) to give the title compound, 2- [ 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetoxy ] -7-azaspiro [3.5] nonane-7-carboxylic acid as a pale yellow liquid (12.42 g, 12.42% yield).
1H NMR(400MHz，CDCl3)7.21(m，1H)，7.11(m，1H)，6.97(m，1H)，5.08(m，1H)，3.96(s，1H)，3.35-3.26(m，4H)，2.85-2.73(m，1H)，2.39(m，1H)，2.31(m，1H)，1.90(m，1H)，1.82(m，1H)，1.61-1.48(m，12H)，1.45(s，9H).
The fourth step: 7-Azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (12E)
7-azaspiro[3.5]nonan-2-yl 2-cyclopentyl-2-hydroxy-2-(2-thienyl)acetate
Figure GPA0000234146600000662
Tert-butyl 2- [ 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetoxy ] -7-azaspiro [3.5] nonane-7-carboxylate (12D) (0.55g, 1.22mmol) was dissolved in 1, 4-dioxane (20m L) and reacted at room temperature for 1 hour with excess hydrochloric acid gas, after completion of the reaction, saturated aqueous sodium bicarbonate (50m L) was added, extraction was performed with dichloromethane (100m L× 2), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (12E) as a colorless liquid (0.40g, 93.6% yield).
1H NMR(400MHz，DMSO-d6)7.38(m，1H)，7.08(m，1H)，6.96(m，1H)，5.93(s，1H)，4.94(m，1H)，2.78-2.68(m，1H)，2.63-2.52(m，4H)，2.24(m，2H)，1.70(m，2H)，1.58-1.39(m，12H).
LCMS m/z＝350.2[M+1].
The fifth step: [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (12F)
[7-[3-(2-chloro-4-formyl-5-methoxy-anilino)-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-(2-thienyl)acetate
Figure GPA0000234146600000671
N- (2-chloro-4-formyl-5-methoxy-phenyl) prop-2-enamide (1E) (0.247g, 2.03mmol) was dissolved in 2-methyltetrahydrofuran (30m L), 7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (12E) (0.300g, 0.858mmol) was added, triethylamine (0.174g, 1.72mmol) was added, microwave reaction was performed at 100 ℃ for 1 hour, the reaction liquid was cooled to room temperature, and after concentration under reduced pressure, the residue was purified by silica gel column chromatography (eluent methanol: dichloromethane (v/v) ═ 1: 99 to 1: 49) to give the title compound [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (69.350 g, 0.69 g yield as a yellow solid).
1H NMR(400MHz，DMSO-d6)10.79(s，1H)，10.18(s，1H)，8.24(s，1H)，7.70(s，1H)，7.39(m，1H)，7.09(m，1H)，6.97(m，1H)，5.94(s，1H)，5.03-4.93(m，1H)，3.89(s，3H)，2.80-2.70(m，1H)，2.62(s，4H)，2.41(d，3H)，2.34-2.20(m，3H)，1.82-1.69(m，2H)，1.61(m，4H)，1.55-1.28(m，8H).
LCMS m/z＝589.1[M+1].
And a sixth step: [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (12G)
[7-[3-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-2-chloro-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-(2-thienyl)acetate
Figure GPA0000234146600000672
[7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (12F) (0.400G, 0.679mmol) was dissolved in a mixed solution of dichloromethane (10m L) and methanol (10m L), 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.318G, 0.951mmol) was added, reaction was carried out at room temperature for 30 minutes, sodium triacetoxyborohydride (0.432G, 2.04mmol) was added, reaction was carried out at room temperature for 3 hours, dichloromethane (20m L) and saturated sodium bicarbonate solution (20m L) were added to the reaction solution, the fractions were extracted with dichloromethane (20m L× 1) for aqueous phase, the organic phases were combined, dried without water, the reaction solution was purified by silica-column chromatography (1 v) with ethyl tert-butyl-2-phenyl ethyl-7-azaspiro [ 3-2-oxo-propyl ] -7-azaspiro [ 3-yl ] 2-cyclopentyl ]2- (2-thienyl ] ethyl-1G, the title compound was purified as a pale yellow solid (0.7-7-azaspiro [ 10G, 10m L) eluent, 2-ethyl tert-butyl-ethyl ether (0.7-tert-butyl-2-ethyl-butyl-7-ethyl-2-ethyl-7-ethyl-1G, 2-7-ethyl-1H-7-ethyl.
LCMS m/z＝454.1[(M+2)/2].
The seventh step: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxyanilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate; bitrifluoroacetate salt (Compound 12)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-(2-thienyl)acetate，；ditrifluoroacetate
Figure GPA0000234146600000681
Dissolving [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] -2-cyclopentyl-2-hydroxy-2- (2-thienyl) acetate (12G) (0.275G, 0.303mmol) in dichloromethane (15m L), adding triethylamine trihydrofluoride (0.488G, 3.03mmol) to the reaction mixture, reacting at room temperature for 24 hours, adding water (20m L) and dichloromethane (20m L) to the reaction mixture, adjusting pH to 12 with 3% sodium hydroxide solution, extracting the separated layers, extracting the aqueous phase with dichloromethane (20m L×) and combining the organic phases, washing with saturated aqueous sodium chloride solution (20m L×) and drying anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, eluting with a column chromatography to obtain crude compound (0.5% ethyl-2-7-2-ethyl-2-phenyl) and purifying the crude compound with a eluent (0.5% ethyl-2-phenyl-ethyl-2-phenyl) to obtain crude compound, eluting with a gradient of ethyl-2-hydroxy-2-cyclopentyl-2-hydroxy-2-thienyl-2-hydroxy-2-thienyl-phenyl-ethyl acetate (0.5-phenyl) at room temperature, eluting, and a gradient at room temperature, and a flow rate of 0.488 min, and a gradient at room temperature for 24 hours at room temperature, and a flow rate of 0.488 hours, and a flow rate of 0% for 24 hours, separating and a flow rate of 0.488 minutes, and a flow rate of 0.7 minutes, and a flow rate of 0% for 24 hours, and a flow.
1H NMR(400MHz，CD3OD)7.22(d，1H)，6.88(s，1H)，6.56(s，1H)，6.44-6.33(m，2H)，6.23(s，1H)，6.13(d，1H)，6.08-6.04(m，1H)，5.70(d，1H)，4.50(m，1H)，4.23-4.13(m，1H)，3.36(m，2H)，2.95(s，3H)，2.70-2.57(m，4H)，2.38-2.32(m，1H)，2.27(m，1H)，2.16(m，4H)，1.96(m，1H)，1.62(m，1H)，1.47(m，1H)，1.08(m，6H)，0.82-0.53(m，8H).
LCMS m/z＝397.3[(M+2)/2].
Example 13: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate; bitrifluoroacetate salt (Compound 13)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate；ditrifluoroacetic acid
Figure GPA0000234146600000691
Figure GPA0000234146600000701
The first step is as follows: 2-oxo-2-phenyl-acetic acid (13B)2-oxo-2-phenyl-acetic acid
Figure GPA0000234146600000702
Methyl 2-oxo-2-phenyl-acetate (13A) (10g, 60.92mmol) was dissolved in water (50M L), sodium hydroxide (4.9g, 121.8mmol) was added and the reaction was allowed to react at room temperature for 2 hours the reaction was extracted with dichloromethane (50M L× 2), the aqueous phase was adjusted to pH 3 with 4M aqueous hydrochloric acid and extracted with dichloromethane (100M L× 3), the organic phases were combined and concentrated under reduced pressure to give the title compound 2-oxo-2-phenyl-acetic acid (13B) as a white solid (9g, 98.41% yield).
1H NMR(400MHz，DMSO-d6)7.95-7.88(m，2H)，7.78-7.71(m，1H)，7.64-7.55(m，2H).
LCMS m/z＝149.1[M-1]。
The second step is that: 2- (2-oxo-2-phenyl-acetyl) oxy-7-azaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (13C)
tert-butyl 2-(2-oxo-2-phenyl-acetyl)oxy-7-azaspiro[3.5]nonane-7-carboxylate
Figure GPA0000234146600000703
Dissolving 2-oxo-2-phenyl-acetic acid (13B) (4.0g, 27mmol) in dichloromethane (100m L), adding oxalyl chloride (6.8g, 53mmol), dropwise adding N, N-dimethylformamide, reacting at room temperature for 1 hour, concentrating the reaction solution under reduced pressure to obtain reaction solution 3. dissolving tert-butyl 2-hydroxy-7-azaspiro [3.5] nonane-7-carboxylate (1B) (4.8g, 20mmol) in dichloromethane (100m L), adding triethylamine (11g, 110mmol), adding reaction solution 3 under ice bath, reacting at room temperature for 1 hour, adding dichloromethane (50m L) and water (50m L) to the reaction solution, extracting the aqueous phase with dichloromethane (50m L× 1), combining the organic phases, drying over anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and purifying the residue by silica gel column chromatography (ethyl acetate: petroleum ether (v/v) ═ 0: 1 to 1: 9) to obtain the title compound [2- (2-oxo-phenyl-nonane-7-carboxylic acid (0.5g, 6g) tert-butyl azaspiro [ 3-azaspiro [3 ] nonane (yield).
1H NMR(400MHz，CDCl3)8.00(d，2H)，7.70-7.62(m，1H)，7.52(t，2H)，5.35-5.28(m，1H)，3.42-3.34(m，2H)，3.34-3.26(m，2H)，2.54-2.41(m，2H)，2.07-1.98(m，2H)，1.64-1.53(m，4H)，1.50-1.37(m，9H).
LCMS m/z＝396.2[M+23]。
The third step: 2- (2-cyclopentyl-2-hydroxy-2-phenyl-acetyl) oxy-7-azaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (13D)
tert-butyl 2-(2-cyclopentyl-2-hydroxy-2-phenyl-acetyl)oxy-7-azaspiro[3.5]nonane-7-carboxylate
Figure GPA0000234146600000711
Tert-butyl 2- (2-oxo-2-phenyl-acetyl) oxy-7-azaspiro [3.5] nonane-7-carboxylate (13C) (1.0g, 11mmol) was dissolved in tetrahydrofuran (100M L), 2.0M tetrahydrofuran solution of cyclopentyl magnesium bromide (8M L, 16mmol) was added at-40 ℃, the reaction was gradually warmed to room temperature for 2 hours, a saturated ammonium chloride solution (60M L) was added to the reaction mixture, ethyl acetate (50M L) was added, the aqueous phase was extracted with ethyl acetate (50M L× 1), 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 (ethyl acetate: petroleum ether (v/v): 0: 1 to 1: 19) to give the title compound 2- (2-cyclopentyl-2-hydroxy-2-phenyl-acetyl) oxy-7-azaspiro [3.5] nonane-7-carboxylate (13D) as a colorless oil (yield, 51.4%).
1H NMR(400MHz，CDCl3)7.68-7.42(m，2H)，7.38-7.29(m，2H)，7.29-7.23(m，1H)，5.08-4.95(m，1H)，3.39-3.17(m，4H)，2.97-2.82(m，1H)，2.44-2.20(m，2H)，1.90-1.80(m，1H)，1.79-1.54(m，7H)，1.53-1.47(m，4H)，1.45-1.43(m，9H)，1.38-1.30(m，2H).
LCMS m/z＝466.3[M+23]。
The fourth step: 7-Azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E)
7-azaspiro[3.5]nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000721
Tert-butyl 2- (2-cyclopentyl-2-hydroxy-2-phenyl-acetyl) oxy-7-azaspiro [3.5] nonane-7-carboxylate (13D) (2.4g, 5.4mmol) was dissolved in dichloromethane (20m L), trifluoroacetic acid (6.2g, 54mmol) was added and reacted at room temperature for 3 hours, aqueous ammonia was added to the reaction solution to adjust pH to 10, dichloromethane (50m L) and water (50m L) were added, the aqueous phase was extracted with dichloromethane (20m L× 1), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E) as a yellow oil (1.7g, 91% yield).
1H NMR(400MHz，CDCl3)7.66-7.40(m，2H)，7.38-7.29(m，2H)，7.29-7.24(m，1H)，5.07-4.95(m，1H)，3.03-2.84(m，4H)，2.51-2.18(m，2H)，2.00-1.27(m，15H).
LCMS m/z＝344.2[M+1]。
The fifth step: [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F)
[7-[3-(2-chloro-4-formyl-5-methoxy-anilino)-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000722
7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E) (0.400g, 1.16mmol) was dissolved in 2-methyltetrahydrofuran (10m L), N- (2-chloro-4-formyl-5-methoxy-phenyl) propyl-2-enamide (1E) (0.307g, 1.28mmol) was added, triethylamine (0.236g, 2.33mmol) was added, microwave reaction was performed at 100 ℃ for 1 hour, the reaction liquid was cooled to room temperature, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v/v): 1 to 1: 0) to give the title compound [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F) in a yellow solid yield (0.73 g, 0.73%).
1H NMR(400MHz，CDCl3)10.93(s，1H)，10.32(s，1H)，8.34(d，1H)，7.81(d，1H)，7.67-7.41(m，2H)，7.38-7.30(m，2H)，7.29-7.23(m，1H)，5.08-4.98(m，1H)，3.93(s，3H)，2.98-2.85(m，1H)，2.74-2.55(m，4H)，2.47-2.23(m，4H)，1.91-1.41(m，14H)，1.39-1.29(m，2H).
LCMS m/z＝583.3[M+1]。
And a sixth step: [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13G)
[7-[3-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-2-chloro-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000731
[7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F) (0.430G, 0.737mmol) was dissolved in methanol (10m L), 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.296G, 0.885mmol) was added, anhydrous zinc chloride (0.402G, 2.95mmol) was added, reaction was carried out at 55 ℃ for 1 hour, sodium cyanoborohydride (0.139G, 2.21mmol) was added, reaction was carried out at 55 ℃ for 2 hour, dichloromethane (50m L) and saturated sodium bicarbonate solution (20m L) were added to the extraction aqueous phase with dichloromethane (30m L× 1), the organic phase was combined, the filtrate was filtered over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure to give the title compound [ 3-oxo-propyl ] -7-azaspiro [ 3-2-yl ] 2-cyclopentyl-hydroxy-2-quinolin-2-phenyl-one (0.296G, 0.885 mmol).
LCMS m/z＝451.3[(M+2)/2]。
The seventh step: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate; bitrifluoroacetate salt (Compound 13)
[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate；ditrifluoroacetic acid
Figure GPA0000234146600000741
[7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13G) (0.450G, 0.499mmol) was dissolved in tetrahydrofuran (5m L), triethylamine trihydrofluoride (0.805G, 4.99mmol) was added, TFA was reacted at room temperature for 24 hours, 10% (v/v) methanol/dichloromethane solution (50m L) was added to the reaction solution, the pH was adjusted to about 8 by adding saturated aqueous sodium bicarbonate solution, the aqueous phase was extracted with 10% (v/v) methanol/dichloromethane solution (50m L×), the organic phases were combined, washed with saturated brine (20m L×) to obtain a filtrate, the filtrate was filtered under reduced pressure, the column chromatography conditions of 0.7% ethyl-2-phenyl-2-ethyl-2-phenyl-2-yl ] acetate, the title compound was purified by column chromatography (0.05-2-ethyl-2-phenyl-ethyl-2-phenyl-2-yl) and the eluent was purified under reduced pressure, the conditions of title compound under reduced pressure of a-2-7-ethyl-7% yield of a-7-phenyl-7-TFA-7% eluent under the title 1-7% eluent under the flow rate of the title 1-7% eluent under the title TFA-7% of the title TFA gradient.
1H NMR(400MHz，CD3OD)8.16(d，1H)，7.79(s，1H)，7.64(d，2H)，7.49-7.42(m，1H)，7.40-7.22(m，4H)，7.05(d，1H)，6.62(d，1H)，5.45-5.35(m，1H)，5.10-4.96(m，1H)，4.28(q，2H)，33.82(s，3H)，3.62-3.40(m，4H)，3.30-3.15(m，2H)，3.13-2.86(m，5H)，2.60-2.24(m，2H)，2.03-1.78(m，6H)，1.76-1.30(m，8H).
19F NMR(376MHz，CD3OD)-75.35.
LCMS m/z＝394.3[(M+2)/2]。
Example 14: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate; isomer 1 of ditrifluoroacetate salt (compound 13)
Isomer 1 of[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate；ditrifluoroaceticacid
Figure GPA0000234146600000751
The first step is as follows: isomer 1 of 7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E)
Isomer 1 of 7-azaspiro[3.5]nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000752
Reacting 2- (2-cyclopentyl-2-hydroxy-2-phenyl-acetyl) oxy-7-azaspiro [3.5]Nonane-7-carboxylic acid tert-butyl ester (13D) (13g, 29.31mmol) was dissolved in dichloromethane (50m L), trifluoroacetic acid (33.41g, 293.1mmol) was added, the reaction was carried out at room temperature for 3 hours, ammonia water was added to the reaction solution to adjust the pH to 10, dichloromethane (50m L) and water (50m L) were added, extraction was carried out, the aqueous phase was extracted with dichloromethane (50m L× 1), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtration was carried out, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by chromatography under the conditions of Instrument: Shimadzu L C-20AP Prep HPLC(PrepL-GB)；Column：ChiralPak AY，300×50mm，10u；Mobile phase：A for Heptane(0.1％NH3H2O) and B for Ethanol, Gradient: B25%, Flow rate: 80m L/min, Columnteroperation: R.T. to give the title compound 7-azaspiro [3.5]Nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E) isomer 1, yellow oil (2.7g, 27% yield), with a peak time for 13E isomer 1 less than that for 13E isomer 2.
1H NMR(400MHz，CDCl3)7.65-7.40(m，2H)，7.37-7.29(m，2H)，7.29-7.23(m，1H)，5.05-4.95(m，1H)，2.96-2.88(m，1H)，2.88-2.72(m，4H)，2.46-2.20(m，2H)，1.91-1.79(m，1H)，1.79-1.40(m，12H)，1.37-1.30(m，1H).
LCMS m/z＝344.3[M+1]。
The second step is that: isomer 1 of [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F)
Isomer 1 of[7-[3-(2-chloro-4-formyl-5-methoxy-anilino)-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000761
Isomer 1(0.600g, 1.75mmol) of 7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E) was dissolved in 2-methyltetrahydrofuran (10m L), N- (2-chloro-4-formyl-5-methoxy-phenyl) propyl-2-enamide (1E) (0.460g, 1.92mmol) was added, triethylamine (0.354g, 3.49mmol) was added, microwave reaction was performed at 100 ℃ for 1 hour, the reaction liquid was cooled to room temperature, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v/v) ═ 1: 1 to 1: 0) to give the title compound [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-phenyl-acetate (1.740 g, 13.72 g, yield as a yellow solid).
1H NMR(400MHz，CDCl3)10.29(d，1H)，8.33(d，1H)，7.81(d，1H)，7.65-7.40(m，2H)，7.37-7.29(m，2H)，7.29-7.24(m，2H)，5.07-4.95(m，1H)，3.93(s，3H)，2.97-2.83(m，1H)，2.68(br，4H)，2.48(br，2H)，2.42-2.34(m，1H)，2.32-2.23(m，1H)，1.92-1.80(m，1H)，1.80-1.65(m，7H)，1.65-1.30(m，8H).
LCMS m/z＝583.3[M+1]。
The third step: isomer 1 of [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13G)
Isomer 1 of[7-[3-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-2-chloro-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000771
Isomer 1(0.740G, 1.27mmol) of [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F) is dissolved in methanol (10m L), 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.425G, 1.27mmol) is added, anhydrous zinc chloride (0.692G, 5.08mmol) is added, reaction is carried out at 55 ℃ for 1H sodium cyanoborohydride (0.239G, 3.81mmol) is added, reaction is carried out at 55 ℃ for 2H dichloromethane (50m L) is added to the reaction solution, saturated sodium bicarbonate solution (20m L) is added, extraction of the aqueous phase is carried out with dichloromethane (30m L× 2), the combined organic phase is filtered, the yellow isomer 1G of [ (3-chloro-2-oxo-2-propyl ] -7-azaspiro [ 3-2-yl ] 2-phenyl-acetate (13F) is concentrated to give the title compound as yellow solid (1G, yield- [ [ - [ 2-hydroxy-2-hydroxy-2-quinolin-2-oxo-2-one (0.7H-2-yl).
LCMS m/z＝451.3[(M+2)/2]。
The fourth step: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate; isomer 1 of ditrifluoroacetate salt (compound 13)
Isomer 1 of[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate；ditrifluoroaceticacid
Figure GPA0000234146600000781
Isomer 1(1.0G, 1.1mmol) of [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13G) is dissolved in tetrahydrofuran (5m L), triethylamine trihydrofluoride salt (1.8G, 11mmol) is added, reaction is carried out at room temperature for 24 hours, 10% (v/v) methanol/dichloromethane solution (50m L) is added to the reaction solution, pH is adjusted to about 8 by adding saturated aqueous sodium bicarbonate solution, extraction is carried out with 10% (v/v) methanol/dichloromethane solution (50m L×), the combined organic phases are extracted with saturated sodium bicarbonate (20m L×), the filtrate is washed with anhydrous sodium bicarbonate solution, the filtrate is filtered under reduced pressure to obtain a filtrate, the title compound is purified by a column chromatography, the procedure is carried out under reduced pressure, the conditions of 0.7-5% eluent of ethyl-phenyl-2-ethyl-2-acetate (13G) and the yield of the title compound is reduced pressure, the concentration is 0.1 min, the title compound is obtained, the product is obtained, the procedure is carried out as a, the procedure of triethylamine trihydrogen phase after the procedure is carried out as a procedure of triethylamine trihydrogen concentration, the procedure is carried out as a procedure of triethylamine trihydrogen concentration conditions of triethylamine trihydrogen phase, the procedure of triethylamine trihydrogen phase after 1 min, the procedure of triethylamine trihydro.
1H NMR(400MHz，CD3OD)8.13(d，1H)，7.77(s，1H)，7.62(d，2H)，7.46(s，1H)，7.37-7.21(m，4H)，7.03(d，1H)，6.60(d，1H)，5.40(dd，1H)，5.09-4.94(m，1H)，4.26(q，2H)，3.89-3.77(m，3H)，3.62-3.40(m，4H)，3.29-3.22(m，1H)，3.21-3.12(m，1H)，3.11-2.86(m，5H)，2.60-2.40(m，1H)，2.40-2.22(m，1H)，2.00-1.80(m，6H)，1.71-1.53(m，5H)，1.52-1.40(m，1H)，1.39-1.26(m，2H).
19F NMR(376MHz，CD3OD)-75.43.
LCMS m/z＝394.4[(M+2)/2]。
Example 15: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate; isomer 2 of ditrifluoroacetate salt (compound 13)
Isomer 2 of[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate；ditrifluoroaceticacid
Figure GPA0000234146600000791
The first step is as follows: isomer 2 of 7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E)
Isomer 2 of 7-azaspiro[3.5]nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000792
Reacting 2- (2-cyclopentyl-2-hydroxy-2-phenyl-acetyl) oxy-7-azaspiro [3.5]Tert-butyl nonane-7-carboxylate (13D) (13g, 29.31mmol) was dissolved in dichloromethane (50m L), and trifluoroacetic acid (33.41g, 293.1mmol) was added to react at room temperature for 3 hours.The reaction solution was adjusted to pH 10 by adding aqueous ammonia, dichloromethane (50m L) and water (50m L) were added, extraction was carried out, the aqueous phase was extracted with dichloromethane (50m L× 1), 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 resolved under the conditions of Instrument: Shimadzu L C-20AP Prep HP L C (Prep L-GB), Column: ChiralPak AY, 300 × 50mm, 10u, Mobile phase: A for Heptane (0.1% NH)3H2O) and B forEthanol, Gradient: B25%, Flow rate: 80m L/min, Column temperature: R.T. to give the title compound 7-azaspiro [3.5]Isomer 2 of nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E), yellow oil (2.50g, 25% yield).
1H NMR(400MHz，CDCl3)9.21(s，1H)，7.62(dd，2H)，7.36-7.30(m，2H)，7.29-7.24(m，1H)，5.08-4.96(m，1H)，3.12-2.94(m，4H)，2.94-2.85(m 1H)，2.48-2.37(m，1H)，2.37-2.27(m，1H)，1.93(dd，1H)，1.86-1.74(m，5H)，1.73-1.40(m，6H)，1.40-1.30(m，2H).
LCMS m/z＝344.3[M+1]。
The second step is that: isomer 2 of [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F)
Isomer 2 of[7-[3-(2-chloro-4-formyl-5-methoxy-anilino)-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000801
Isomer 2(0.600g, 1.75mmol) of 7-azaspiro [3.5] nonan-2-yl 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13E) was dissolved in 2-methyltetrahydrofuran (10m L), N- (2-chloro-4-formyl-5-methoxy-phenyl) propyl-2-enamide (1E) (0.460g, 1.92mmol) was added, triethylamine (0.354g, 3.49mmol) was added, microwave reaction was performed at 100 ℃ for 1 hour, the reaction solution was concentrated, the residue was purified by silica gel column chromatography (ethyl acetate: petroleum ether (v/v) ═ 1: 1 to 1: 0), to give the title compound [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F) as a yellow solid (yield, 0.73% yield).
1H NMR(400MHz，CDCl3)10.30(s，1H)，8.34(s，1H)，7.81(s，1H)，7.68-7.62(m，2H)，7.37-7.29(m，2H)，7.29-7.24(m，2H)，5.09-4.94(m，1H)，3.93(s，3H)，2.98-2.84(m，1H)，2.67(s，4H)，2.48(s，2H)，2.42-2.35(m，1H)，2.32-2.22(m，1H)，1.91-1.83(m，1H)，1.80-1.39(m，13H)，1.39-1.29(m，2H).
LCMS m/z＝583.3[M+1]。
The third step: isomer 2 of [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13G)
Isomer 2 of[7-[3-[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-2-chloro-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate
Figure GPA0000234146600000811
Isomer 2(0.752G, 1.29mmol) of [7- [3- (2-chloro-4-formyl-5-methoxy-anilino) -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13F) is dissolved in methanol (10m L), 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.432G, 1.29mmol) is added, anhydrous zinc chloride (0.701G, 5.14mmol) is added, reaction is carried out at 55 ℃ for 1H sodium cyanoborohydride (0.243G, 3.86mmol) is added, reaction is carried out at 55 ℃ for 2H dichloromethane (50m L) is added to the reaction solution, saturated sodium bicarbonate solution (20m L) is added, extraction of the aqueous phase is carried out with dichloromethane (30m L× 2), the organic phase is combined, the organic phase is filtered, the yellow isomer 2-oxo-propyl ] -7-azaspiro [ 3-2-yl ] 2-phenyl-acetate (13F) is concentrated to give the title compound as yellow solid [ (1G, 2-hydroxy-1H-quinolin-2-one (0.432G, 1G- [ [ -2-oxo-2-one (0.29 mmol).
LCMS m/z＝451.4[(M+2)/2]。
The fourth step: [7- [3- [ 2-chloro-4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate; isomer 2 of ditrifluoroacetate salt (compound 13)
Isomer 2 of[7-[3-[2-chloro-4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]-5-methoxy-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-cyclopentyl-2-hydroxy-2-phenyl-acetate；ditrifluoroaceticacid
Figure GPA0000234146600000812
Isomer 2(1.0G, 1.1mmol) of [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy ] -2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] -2-chloro-5-methoxy-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-cyclopentyl-2-hydroxy-2-phenyl-acetate (13G) was dissolved in tetrahydrofuran (5m L), triethylamine trihydrofluoride (1.8G, 11mmol) was added, reaction was performed at room temperature for 24 hours, 10% (v/v) methanol/dichloromethane solution (50m L) was added to the reaction solution, pH was adjusted to about 8 by adding saturated aqueous sodium bicarbonate solution, extraction was performed using 10% (v/v) methanol/dichloromethane solution (50m L×), organic phases were combined, saturated sodium bicarbonate solution (20m L×) was used, filtrate was washed with anhydrous sodium bicarbonate solution, filtered, the filtrate was dried over a column with anhydrous column, the eluent was prepared, the title compound was obtained as a white solid, filtered, the eluent was obtained as a-2-hydroxy-2-phenyl-2-ethyl-2-phenyl-acetate (13G), the title compound, the eluent was purified under the conditions of a-2-cyclopentyl-2-phenyl-2-phenyl-ethyl-acetate (13G-2-phenyl-acetate (13G) and the eluent, the title compound was dissolved in a-2-cyclopentyl-2-phenyl-ethyl-2-phenyl-ethyl-2-ethyl-phenyl-acetate (13G, the concentration of the eluent under the concentration of.
1H NMR(400MHz，CD3OD)8.04(d，1H)，7.70(s，1H)，7.54(d，2H)，7.38(s，1H)，7.29-7.13(m，4H)，6.95(d，1H)，6.52(d，1H)，5.32(dd，1H)，4.98-4.87(m，1H)，4.17(q，2H)，3.77(s，3H)，3.50-3.30(m，4H)，3.20-3.13(m，1H)，3.13-3.04(m，1H)，3.02-2.77(m，5H)，2.50-2.30(m，1H)，2.31-2.17(m，1H)，1.93-1.69(m，6H)，1.65-1.45(m，5H)，1.45-1.32(m，1H)，1.32-1.20(m，2H).
19F NMR(376MHz，CD3OD)-75.43.
LCMS m/z＝394.3[(M+2)/2]。
Example 16: [7- [3- [3- [ [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] phenyl ] carbamoyl ] -N-methyl-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate; bitrifluoroacetate salt (Compound 16)
[7-[3-[3-[[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]phenyl]carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000831
The first step is as follows: 3- (methylamino) benzoic acid methyl ester (16B)
methyl 3-(methylamino)benzoate
Figure GPA0000234146600000832
Methyl 3-aminobenzoate (16A) (10.0g, 66.2mmol) was dissolved in N, N-dimethylformamide (150m L), potassium carbonate (13.7g, 99.2mmol) was added, methyl iodide (9.39g, 66.2mmol) was further slowly dropped, reaction was performed at room temperature for 3 hours after the addition was completed, filtration was performed, the residue was washed with ethyl acetate (50m L), the filtrate was collected, water (100m L) was added, extraction was performed with ethyl acetate (200m L× 2), the organic phases were combined, washing was performed with a saturated sodium chloride solution (100m L× 2), drying was performed with anhydrous sodium sulfate, filtration was performed, and the residue was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: ethyl acetate (v/v): 100: 0 to 97: 3) to obtain the title compound methyl 3- (methylamino) benzoate (16B), a pale yellow oil (3.85g, yield 35.2%).
1H NMR(400MHz，CDCl3)7.29(d，1H)，7.20-7.17(m，1H)，7.15(t，1H)，6.70(m，1H)，3.81(s，3H)，2.78(s，3H).
LCMS m/z＝166.2[M+1]。
The second step is that: 3- [ methyl (prop-2-enoyl) aminobenzoic acid methyl ester (16C)
methyl 3-[methyl(prop-2-enoyl)amino]benzoate
Figure GPA0000234146600000841
Methyl 3- (methylamino) benzoate (16B) (0.500g, 3.03mmol) was dissolved in dichloromethane (5m L), sodium bicarbonate (0.509g, 6.05mmol) was added, the mixture was cooled to 0 ℃ in an ice bath under nitrogen protection, acryloyl chloride (0.411g, 4.54mmol) was added dropwise, after completion of the addition, the mixture was warmed to room temperature to react for 30 minutes, dichloromethane (20m L× 1) was added for extraction, the mixture was washed with a saturated sodium bicarbonate solution (10m L× 2), the organic phases were combined, the organic phases were washed with a saturated sodium chloride solution (10m L× 1), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (eluent petroleum ether: ethyl acetate (v/v) ═ 100: 0 to 4: 1) to give the title compound methyl 3- [ methyl (prop-2-enoyl) aminobenzoate (16C), a colorless oil (0.9 g, yield 85.7%).
1H NMR(400MHz，CDCl3)8.01(d，1H)，7.87(t，1H)，7.50(t，1H)，7.40-7.36(m，1H)，6.39(m，1H)，6.04(m，1H)，5.55(m，1H)，3.94(s，3H)，3.38(s，3H).
LCMS m/z＝220.2[M+1]。
The third step: 3- [ methyl (prop-2-enoyl) amino ] benzoic acid (16D)
3-[methyl(prop-2-enoyl)amino]benzoic acid
Figure GPA0000234146600000842
Methyl 3- [ methyl (prop-2-enoyl) aminobenzoate (16C) (0.550g, 2.51mmol) was dissolved in tetrahydrofuran (17M L), 1.0M aqueous lithium hydroxide (0.409g, 17.1mmol) was added dropwise with ice-bath cooling, and after completion of addition, reaction was carried out in ice-bath for 1 hour, pH was adjusted to 1 with 10% hydrochloric acid solution under ice-bath, extraction was carried out with ethyl acetate (20M L× 2), the organic phases were combined, washed with saturated sodium chloride solution (10M L× 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 3- [ methyl (prop-2-enoyl) amino ] benzoic acid (16D), a white solid (0.500g, 97.1% yield).
1H NMR(400MHz，CDCl3)8.02(d，1H)，7.88(t，1H)，7.48(t，1H)，7.37(d，1H)，6.35(m，1H)，5.99(m，1H)，5.51(m，1H)，3.34(s，3H).
LCMS m/z＝206.2[M+1]。
The fourth step: 3- [3- [2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] nonan-7-yl ] propionyl-methyl-amino ] benzoic acid (16E)
3-[3-[2-[2-hydroxy-2，2-bis(2-thienyl)acetyl]oxy-7-azaspiro[3.5]nonan-7-yl]propanoyl-methyl-amino]benzoic acid
Figure GPA0000234146600000851
3- [ methyl (prop-2-enoyl) amino ] benzoic acid (16D) (0.400g, 1.95mmol) was dissolved in 2-methyltetrahydrofuran (10m L), 7-azaspiro [3.5] nonan-2-yl 2-hydroxy-2, 2-bis (2-thienyl) acetate (1D) (0.709g, 1.95mmol) and triethylamine (0.394g, 3.90mmol) were added, and after completion of the addition, the reaction mixture was reacted at 100 ℃ for 1 hour, and after concentration of the reaction mixture under reduced pressure, the residue was chromatographed on a silica gel column (eluent dichloromethane: methanol (v/v) ═ 100: 0 to 9: 1) to give the title compound 3- [2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl ] oxy-7-azaspiro [3.5] nonan-7-yl ] propionyl-methyl-amino ] benzoic acid (16E) as a yellow solid (0.500g, 45.1%).
1H NMR(400MHz，DMSO-d6)7.87(d，1H)，7.76(d，1H)，7.50(d，2H)，7.47(d，1H)，7.45(d，1H)，7.26(s，1H)，7.08(d，1H)，7.07(d，1H)，6.98(d，1H)，6.97(d，1H)，5.04-4.96(m，1H)，3.17(s，3H)，2.73(m，2H)，2.24-2.14(m，5H)，2.14-2.07(m，2H)，1.71-1.67[m，2H)，1.46(s，2H)，1.41(s，2H)，1.24(s，1H).
LCMS m/z＝569.3[M+1]。
The fifth step: [7- [3- [3- [ [4- (1, 3-dioxolan-2-yl) phenyl ] carbamoyl ] -N-methylaniline ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (16F)
[7-[3-[3-[[4-(1，3-dioxolan-2-yl)phenyl]carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000852
3- [3- [2- [ 2-hydroxy-2, 2-bis (2-thienyl) acetyl]Oxy-7-azaspiro [3.5]Nonan-7-yl]Propionyl-methyl-amino]Benzoic acid (16E) (1.50g, 2.64mmol) was dissolved in dichloromethane (20m L) and 4- (1, 3-dioxolan-2-yl) aniline was added
Figure GPA0000234146600000853
(0.871g, 5.28mmol), HATU (1.30g, 3.43mmol), cooling to 0 ℃ in an ice bath, adding diisopropylethylamine (1.70g, 13.2mmol) dropwise, after the addition, naturally warming to room temperature to react for 3 hours, adding water (10m L), dichloromethane (20m L), separating, washing the organic phase with a saturated sodium chloride solution (10m L× 1), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and purifying the resulting crude product with a silica gel column (eluent:70% (v/v) ethyl acetate/petroleum ether-2% (v/v) methanol/dichloromethane-4% (v/v) methanol/dichloromethane to obtain the title compound [7- [3- [3- [ [4- (1, 3-dioxolan-2-yl) phenyl ] ethyl acetate/petroleum ether]Carbamoyl radical]-N-methylaniline]-3-oxo-propyl]-7-azaspiro [3.5]Nonan-2-yl]2-hydroxy-2, 2-bis (2-thienyl) acetate (16F), light yellow solid (0.820g, 43.4% yield).
1H NMR(400MHz，CDCl3)7.87(d，1H)，7.80-7.73(m，3H)，7.53(t，1H)，7.48(d，2H)，7.34(d，1H)，7.28(m，2H)，7.15(m，2H)，6.97(m，2H)，5.80(s，1H)，5.14-5.05(m，1H)，4.76-4.60(m，1H)，4.15-4.10(m，2H)，4.05-4.00(m，2H)，3.28(s，3H)，2.80(t，2H)，2.50-2.34(m，5H)，2.34-2.26(m，2H)，1.84-1.81(m，2H)，1.67-1.57(m，4H).
LCMS m/z＝716.3[M+1]。
And a sixth step: [7- [3- [3- [ (4-formylphenyl) carbamoyl ] -N-methylaniline ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (16G)
[7-[3-[3-[(4-formylphenyl)carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000861
[7- [3- [3- [ [4- (1, 3-dioxolan-2-yl) phenyl ] carbamoyl ] -N-methylaniline ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (16F) (0.820G, 1.15mmol) was dissolved in acetonitrile (8M L), and 3M aqueous hydrochloric acid (10M L) was added under ice-cooling, followed by reaction for 3 hours on ice, extraction with dichloromethane (20M L× 2), organic phases were combined, the organic phase was washed with a saturated sodium chloride solution (10M L× 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound [7- [3- [3- [ (4-formylphenyl) carbamoyl ] -N-methylaniline ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (16.82G, a crude product (16.82% G) as an impure solid for the next step.
1H NMR(400MHz，CDCl3)9.94(s，1H)，8.05(s，2H)，7.93(d，1H)，7.89(d，2H)，7.84(s，1H)，7.57(t，1H)，7.35(d，1H)，7.29(m，2H)，7.15(m，2H)，6.97(m，2H)，5.18-5.09(m，1H)，4.62(s，1H)，3.29(s，3H)，3.06-2.98(m，2H)，2.75-2.61(m，3H)，2.40-2.31(m，2H)，1.91-1.85(m，3H)，1.80-1.70(m，5H).
LCMS m/z＝672.2[M+1]。
The seventh step: [7- [3- [3- [ [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] phenyl ] carbamoyl ] -N-methyl-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (16H)
[7-[3-[3-[[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]phenyl]carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000871
[7- [3- [3- [ (4-formylphenyl) carbamoyl ] -N-methylaniline ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (16G) (0.600G, 0.893mmol) was dissolved in dry methanol (20m L), 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.299G, 0.893mmol) and dry zinc chloride (0.487G, 3.57mmol) were added after reaction for 1 hour at 50 ℃ sodium cyanoborohydride (0.281G, 4.47mmol) was added, reaction was continued for 1 hour at 50 ℃ water (50m L), dichloromethane (100m L× 3) was extracted, the organic phases were combined, the organic phase was washed with saturated sodium chloride solution 50m L× 1%, the filtrate was washed with dry anhydrous butyl-2-hydroxy-2, 2-di (2-thienyl) acetate (2H-quinol-2-oxo-propyl ] -7-azaspiro [ 3-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (7-ethyl-2-phenyl) was concentrated under reduced pressure to give crude product (7-azaspiro [ 3-ethyl-2-ethyl-2-oxo-2-oxo-2-yl) as a yellow solid, 7-2-yl).
LCMS m/z＝495.8[M/2+1]。
Eighth step: [7- [3- [3- [ [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] phenyl ] carbamoyl ] -N-methyl-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate; bitrifluoroacetate salt (Compound 16)
[7-[3-[3-[[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]phenyl]carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000881
Dissolving [7- [3- [3- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] methyl ] phenyl ] carbamoyl ] -N-methyl-anilino ] -3-oxo-propyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (16H) (0.600g, 0.606mmol) in tetrahydrofuran (10m L), adding triethylamine trihydrofluoride (0.391g, 2.42mmol) to react overnight at room temperature, pouring off the reaction clear solution, washing the remaining viscous substance with tetrahydrofuran (20m L), pouring off the clear solution, pouring off the viscous substance, adjusting the viscous substance with a mixed solution of methanol/dichloromethane (v ═ 1/9), separating the saturated aqueous sodium bicarbonate solution, adjusting the pH to an aqueous layer, then using a mixed solution of methanol/dichloromethane (v/v ═ 1/9), filtering the obtained crude compound with dichloromethane (3-2- [3- [ [ [ (2R) -2-tert-butyl (dimethyl) silyl ] oxy-2- (8-quinolin-5-yl) ethyl ] 2-yl ] acetate (0.05% of ethyl-2-hydroxy-2-yl) and purifying the obtained crude compound with a gradient, after elution under a gradient of 0.7-5-2-5-2-hydroxy-2, after a gradient, after a.
1H NMR(400MHz，CD3OD)8.19(d，1H)，8.01(d，1H)，7.92(s，1H)，7.83(d，2H)，7.67(t，1H)，7.58(d，1H)，7.51(d，2H)，7.39-7.34(m，2H)，7.27(d，1H)，7.12(s，2H)，7.02(d，1H)，6.99-6.94(m，2H)，6.63(d，1H)，5.39(t，1H)，5.16-5.08(m，1H)，4.30(s，2H)，3.51-3.35(m，4H)，3.33(s，3H)，3.22(d，2H)，2.94-2.80(m，2H)，2.61(t，2H)，2.51-2.45(m，1H)，2.36-2.30(m，1H)，1.98-1.88(m，3H)，1.82-1.70(m，3H).
19F NMR(376MHz，CD3OD)-75.44.
LCMS m/z＝876.4[M+1]。
Example 17: [7- [3- [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] aminomethyl ] phenyl ] carbamoyl ] -N-methylaniline ] -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate; bitrifluoroacetate salt (Compound 17)
[7-[3-[4-[[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]phenyl]carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000891
The first step is as follows: 4- (methylamino) benzoic acid methyl ester (17B)
methyl 4-(methylamino)benzoate
Figure GPA0000234146600000892
Methyl 4-aminobenzoate (17A) (10.0g, 66.2mmol) was dissolved in N, N-dimethylformamide (150m L), potassium carbonate (13.7g, 99.2mmol) was added, methyl iodide (9.39g, 66.2mmol) was further slowly dropped, reaction was performed at room temperature for 3 hours after the addition was completed, filtration was performed, the residue was washed with ethyl acetate (50m L), the filtrate was collected, water (100m L) was added, extraction was performed with ethyl acetate (200m L× 2), the organic phases were combined, the organic phase was washed with a saturated sodium chloride solution (100m L× 2), dried over anhydrous sodium sulfate, filtration was performed, and the residue after concentration of the filtrate under reduced pressure was separated and purified by silica gel column chromatography (eluent: ethyl acetate (v/v): 100: 0 to 97: 3) to obtain the title compound methyl 4- (methylamino) benzoate (17B), a white solid (4.80g, yield 43.9%).
1H NMR(400MHz，CDCl3)7.81-7.79(m，1H)，7.79-7.77(m，1H)，6.50-6.48(m，1H)，6.48-6.45(m，1H)，3.77(s，3H)，2.80(s，3H).
LCMS m/z＝166.2[M+1]。
The second step is that: 4- [ methyl (prop-2-enoyl) aminobenzoic acid methyl ester (17C)
methyl 4-[methyl(prop-2-enoyl)amino]benzoate
Figure GPA0000234146600000901
The compound 4- (methylamino) benzoic acid methyl ester (17B) (1.80g, 10.90mmol) was dissolved in dichloromethane (5m L), sodium bicarbonate (1.831g, 21.8mmol) was added, the mixture was cooled to 0 ℃ in an ice bath under nitrogen protection, acryloyl chloride (1.48g, 16.3mmol) was added dropwise, after the addition was completed, the mixture was warmed to room temperature to react for 30 minutes, dichloromethane (20m L× 1) was added to extract, the organic phase was washed with a saturated sodium bicarbonate solution (10m L× 2), a saturated sodium chloride solution (10m L× 1) was washed, anhydrous sodium sulfate was dried, filtration was performed, and the filtrate was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography to purify (eluent petroleum ether: ethyl acetate (v/v) ═ 100: 0 to 4: 1) to obtain the title compound 4- [ methyl (propan-2-enoyl) aminobenzoic acid methyl ester (17C), a colorless oil (1.80g, yield 75.3%).
1H NMR(400MHz，CDCl3)8.10-8.06(m，2H)，7.27(d，2H)，7.24(d，1H)，6.40(m，1H)，6.09(m，1H)，5.57(m，1H)，3.94(s，3H)，3.39(s，3H).
LCMS m/z＝220.2[M+1]。
The third step: 4- [ methyl (prop-2-enoyl) amino ] benzoic acid (17D)
4-[methyl(prop-2-enoyl)amino]benzoic acid
Figure GPA0000234146600000902
Methyl 4- [ methyl (prop-2-enoyl) aminobenzoate (17C) (3.100g, 14.14mmol) was dissolved in tetrahydrofuran (50M L), 1.0M aqueous lithium hydroxide (2.303g, 96.15mmol) was added dropwise with ice-bath cooling, and after completion of addition, reaction was carried out in ice-bath for 1 hour, pH was adjusted to 1 with 10% hydrochloric acid solution under ice-bath, extraction was carried out with ethyl acetate (20M L× 2), the organic phases were combined, washed with saturated sodium chloride solution (10M L× 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 4- [ methyl (prop-2-enoyl) amino ] benzoic acid (17D) as a white solid (2.60g, 89.8% yield).
1H NMR(400MHz，DMSO-d6)13.05(s，1H)，8.02-8.00(m，1H)，8.00-7.97(m，1H)，7.43-7.40(m，1H)，7.40-7.38(m，1H)，6.19(m，1H)，6.12(m，1H)，5.62(m，1H)，3.29(s，3H).
LCMS m/z＝206.2[M+1]。
The fourth step: n- [4- (1, 3-Dioxolan-2-yl) phenyl ] -4- [ methyl (prop-2-enoyl) amino ] benzamide (17E)
N-[4-(1，3-dioxolan-2-yl)phenyl]-4-[methyl(prop-2-enoyl)amino]benzamide
Figure GPA0000234146600000911
Dissolving 4- [ methyl (prop-2-enoyl) amino ] benzoic acid (17D) (0.500g, 2.44mmol) in dichloromethane (10m L), adding 4- (1, 3-dioxolan-2-yl) aniline (0.604g, 3.65mmol), HATU (1.20g, 3.17mmol), cooling in an ice bath to 0 ℃, further adding diisopropylethylamine (1.57g, 12.2mmol), naturally warming to room temperature after completion of the addition, reacting for 3 hours, adding water (10m L) and dichloromethane (20m L), separating, washing the organic phase with a saturated sodium chloride solution (10m L× 1), drying over anhydrous sodium sulfate, filtering, and concentrating the filtrate under reduced pressure to obtain a crude product, purifying with a silica gel column (eluent: 20% (v/v) ethyl acetate/petroleum ether to 70% (v/v) ethyl acetate/petroleum ether) to obtain the title compound N- [4- (1, 3-dioxolan-2-yl) phenyl ] -4- [ methyl (prop-2-enoyl) amino ] benzamide (0.17g, 2.17 g of yellow solid (93%).
LCMS m/z＝353.2[M+1]。
The fifth step: n- (4-formylphenyl) -4- [ methyl (prop-2-enoyl) amino ] benzamide (17F)
N-(4-formylphenyl)-4-[methyl(prop-2-enoyl)amino]benzamide
Figure GPA0000234146600000912
N- [4- (1, 3-Dioxolan-2-yl) phenyl ] -4- [ methyl (prop-2-enoyl) amino ] benzamide (17E) (0.800g, 2.27mmol) was dissolved in acetonitrile (8M L), 3M dilute hydrochloric acid (10M L) was added under cooling in an ice bath, and after completion of the addition, reaction was carried out in an ice bath for 3 hours, extraction was carried out with dichloromethane (20M L× 2), the organic phases were combined, washed with a saturated sodium chloride solution (10M L× 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound N- (4-formylphenyl) -4- [ methyl (prop-2-enoyl) amino ] benzamide (17F) as a crude product as a pale yellow solid (0.560g, yield 80.0%), which was used as an impure in the next step.
1H NMR(400MHz，CDCl3)9.96(s，1H)，7.96(d，2H)，7.94-7.85(m，4H)，7.32(d，2H)，6.40(m，1H)，6.11(m，1H)，5.59(m，1H)，3.40(s，3H).
LCMS m/z＝309.2[M+1]。
And a sixth step: [7- [3- [4- [ (4-formylphenyl) carbamoyl ] -N-methylaniline ] -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (17G)
[7-[3-[4-[(4-formylphenyl)carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000921
N- (4-formylphenyl) -4- [ methyl (prop-2-enoyl) amino ] benzamide (17F) (0.560G, 1.82mmol) was dissolved in 2-methyltetrahydrofuran (10m L), 7-azaspiro [3.5] nonan-2-yl 2-hydroxy-2, 2-bis (2-thienyl) acetate (1D) (0.660G, 1.82mmol) and triethylamine (0.368G, 3.63mmol) were added, and after completion of addition, the reaction mixture was reacted at 100 ℃ for 1 hour, the reaction solution was cooled to room temperature, and after concentration under reduced pressure, the residue was purified by silica gel column chromatography (eluent petroleum ether: ethyl acetate (v/v): 4: 1 to 1: 4) to give the title compound [7- [3- [4- [ (4-formylphenyl) carbamoyl ] -N-methylaniline ] -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-diyl acetate (2-thienyl) in a yield of 650.53G as a pale yellow solid.
LCMS m/z＝672.3[M+1]。
The seventh step: [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] aminomethyl ] phenyl ] carbamoyl ] -N-methylaniline ] -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-bis (2-thienyl) acetate (17H)
[7-[3-[4-[[4-[[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]phenyl]carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate
Figure GPA0000234146600000922
[7- [3- [4- [ (4-formylphenyl) carbamoyl ] -N-methylaniline ] -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (17G) (0.635G, 0.945mmol) was dissolved in dry methanol (20m L), 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.316G, 0.945mmol) and dry zinc chloride (0.515G, 3.78mmol) were added, after reaction at 50 ℃ for 1 hour sodium cyanoborohydride (0.297G, 4.73mmol) was added, reaction was continued at 50 ℃ for 1 hour water (50m L), dichloromethane (100m L× 3) was extracted, the organic phases were combined, the organic phase was extracted with saturated sodium chloride solution (50m L× 1%) and the filtrate was washed with dry sodium tert-butylsilyl-2-hydroxy-2-phenyl-oxo-propyl ] -7-azaspiro [ 3-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (0.8- [ [) was concentrated under reduced pressure to give the crude product of the title compound (1H-2-ethyl-2-quinolino-2-one (7-yl) as a yellow solid which was then filtered, 4- [ [ -2-quinolinone (1G- [ [ -2-one (1G, 0.945mmol) and dried.
LCMS m/z＝495.8[M/2+1]。
Eighth step: [7- [3- [4- [ [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] aminomethyl ] phenyl ] carbamoyl ] -N-methylaniline ] -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate; bitrifluoroacetate salt (Compound 17)
[7-[3-[4-[[4-[[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]methyl]phenyl]carbamoyl]-N-methyl-anilino]-3-oxo-propyl]-7-azaspiro[3.5]nonan-2-yl]2-hydroxy-2，2-bis(2-thienyl)acetate；ditrifluoroaceticacid
Figure GPA0000234146600000931
Dissolving [7- [3- [4- [ [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] aminomethyl ] phenyl ] carbamoyl ] -N-methylaniline ] -3-oxopropyl ] -7-azaspiro [3.5] nonan-2-yl ] 2-hydroxy-2, 2-di (2-thienyl) acetate (17H) (0.600g, 0.606mmol) in tetrahydrofuran (10m L), adding triethylamine trihydrofluoride (0.391g, 2.42mmol) to react overnight at room temperature, pouring off the reaction supernatant, washing the remaining residue with tetrahydrofuran (20m L), pouring off the supernatant, adjusting the pH of the viscous product with a mixed solution of methanol/dichloromethane (v/v ═ 1/9), adding sodium bicarbonate to an alkaline solution, separating the solution, adjusting the pH to alkaline, then using a mixed solution of methanol/dichloromethane (v/v ═ 34) as an aqueous layer, drying the viscous product, eluting with a column (5-5% aqueous layer, purifying the resulting crude product with a-2-ethyl-2-yl-2-yl-2, eluting the resulting crude product with a-2-5% aqueous layer, purifying under a gradient, the gradient of methanol-2-5-7-2-7-2-7-8-7-8-7-8-7-8-7-8.
1H NMR(400MHz，CD3OD)8.15(d，1H)，7.97(d，2H)，7.73(d，2H)，7.44-7.39(m，4H)，7.27(m，2H)，7.17(d，1H)，7.03(t，2H)，6.93(d，1H)，6.89-6.86(m，2H)，6.54(d，1H)，5.35-5.25(m，1H)，5.07-4.98(m，1H)，4.21(s，2H)，3.35-3.22(m，7H)，3.14(d，2H)，2.90-2.72(m，2H)，2.57-2.47(m，2H)，2.45-2.35(m，1H)，2.28-2.20(m，1H)，1.90-1.78(m，3H)，1.75-1.65(m，3H).
19F NMR(376MHz，CD3OD)-75.48.
LCMS m/z＝876.3[M+1]。
Example 18: [7- [ [4- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethylmethylcarbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate; bitrifluoroacetate salt (Compound 18)
[7-[[4-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl-methyl-carbamoyl]phenyl]methyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000941
Figure GPA0000234146600000951
The first step is as follows: 4- [ [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] methyl ] benzoic acid methyl ester (18A)
methyl 4-[[2-[(2-phenylphenyl)carbamoyloxy]-7-azaspiro[3.5]nonan-7-yl]methyl]benzoate
Figure GPA0000234146600000952
7-Azaspiro [3.5] non-2-yl-N- (2-phenylphenyl) carbamate (2D) (1.50g, 4.46mmol) was dissolved in acetonitrile (30m L), N-dimethylformamide (10m L), potassium carbonate (1.85g, 13.4mmol) and methyl 4- (bromomethyl) benzoate (1.02g, 4.46mmol) were added, the temperature was raised to 70 ℃ to react for 2 hours, water (50m L) was added, methylene chloride (100m L× 3) was used for extraction, the organic phases were combined, dried with anhydrous sulfuric acid, filtered, the filtrate was concentrated under reduced pressure, and the residue was chromatographically purified with a silica gel column (eluent dichloromethane: methanol (v/v) ═ 99: 1 to 97: 3) to give the title compound 4- [ [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] methyl ] benzoate (18A) as a white solid in a yield (2.0.6 g, 92.6%).
1H NMR(400MHz，DMSO-d6)8.60(s，1H)，7.96(m，2H)，7.71(s，1H)，7.37(m，10H)，4.72(s，1H)，4.34(s，1H)，3.85(s，3H)，3.47(s，1H)，3.16(m，1H)，2.89(m，1H)，2.17(m，4H)，1.63(m，6H).
LCMS m/z＝485.3[M+1]。
The second step is that: 4- [ [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] methyl ] benzoic acid (18B)
4-[[2-[(2-phenylphenyl)carbamoyloxy]-7-azaspiro[3.5]nonan-7-yl]methyl]benzoic acid
Figure GPA0000234146600000961
Methyl 4- [ [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] methyl ] benzoate (18A) (2.20g, 4.54mmol) was dissolved in tetrahydrofuran (30m L), aqueous sodium hydroxide (7.3g, 61.0mmol) solution (5m L) was added and stirred at room temperature for 2 hours water (50m L) was added, the pH was adjusted to about 2 with 10% aqueous hydrochloric acid, extraction was performed with dichloromethane (100m L× 3), the organic phases were combined, dried with anhydrous sulfuric acid, filtered, and the filtrate was concentrated under reduced pressure to give the title compound 4- [ [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] methyl ] benzoic acid (18B) as a white solid (2.0g, 93.6% yield).
LCMS m/z＝471.3[M+1]。
The third step: [7- [ [4- [2, 2-Dimethylethyl (methyl) carbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate (18C)
[7-[[4-[2，2-dimethoxyethyl(methyl)carbamoyl]phenyl]methyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000962
4- [ [2- [ (2-phenylphenyl) carbamoyloxy ] -7-azaspiro [3.5] non-7-yl ] methyl ] benzoic acid (18B) (2.00g, 4.01mmol) was dissolved in dichloromethane (20m L), N-dimethylformamide (4m L), 2-methoxy-N-methylethylamine (0.531g, 4.45mmol), HATU (2.29g, 6.02mmol) and triethylamine (1.22g, 12.0mmol) were added, reaction was kept at room temperature for 3 hours after completion of addition, dichloromethane (50m L) and water (20m L) were added to the reaction solution, separation was extracted, the organic phase was washed with a saturated aqueous sodium chloride solution (20m L× 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound [7- [ [4- [2, 2-dimethylethyl (methyl) carbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] phenyl ] carbamate (yield: 18.79%).
The fourth step: [7- [ [4- [ methyl (2-oxoethyl) carbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate (18D)
[7-[[4-[methyl(2-oxoethyl)carbamoyl]phenyl]methyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000971
[7- [ [4- [2, 2-dimethylethyl (methyl) carbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate (18C) (1.50g, 2.62mmol) was dissolved in dichloromethane (20m L), p-toluenesulfonic acid (2.26g, 13.1mmol) was added, and reaction was carried out at room temperature for 3 hours, water (20m L) was added, dichloromethane (50m L× 2) was extracted, the organic layers were combined, washed with a saturated aqueous sodium bicarbonate solution (30m L× 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the title compound [7- [ [4- [ methyl (2-oxoethyl) carbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate (18D), a pale yellow solid (1.22g, 88.5% yield).
LCMS m/z＝526.3[M+1]。
The fifth step: [7- [ [4- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethylmethylcarbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate (18E)
[7-[[4-[2-[[(2R)-2-[tert-butyl(dimethyl)silyl]oxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-yl)ethyl]amino]ethyl-methyl-carbamoyl]phenyl]methyl]-7-azaspiro[3.5]nonan-2-yl]N-(2-phenylphenyl)carbamate
Figure GPA0000234146600000972
[7- [ [4- [ methyl (2-oxoethyl) carbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate (18D) (0.200G, 0.381mmol) and 5- [ (1R) -2-amino-1- [ tert-butyl (dimethyl) silyl ] oxymethyl ] -8-hydroxy-1H-quinolin-2-one (1G) (0.127G, 0.381mmol) were dissolved in a mixed solvent of isopropanol/dichloromethane (v/v ═ 1/1, 20m L), glacial acetic acid (1m L) was added, stirring was carried out at room temperature for 1 hour, sodium triacetoxyborohydride (0.162G, 0.761mmol) was then added, dichloromethane (50m L) and water (50m L) were further stirred for 2 hours, the layers were extracted, the organic phase was washed with a saturated aqueous sodium chloride solution (20m 21), anhydrous sodium sulfate was dried, the filtrate was filtered, the residue was subjected to silica gel column chromatography (3G-ethyl-2-phenyl) to obtain a crude product (3.5G-azaspiro [ 3-2-yl ] nonan-2-yl ] as a solid, and the title compound was purified by column chromatography (v [ (-2-ethyl-8-hydroxy-2-8H-quinolino-2-one (0G).
And a sixth step: [7- [ [4- [2- [ [ (2R) -2-hydroxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethylmethylcarbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate; bitrifluoroacetate salt (Compound 18)
[7-[[4-[2-[[(2R)-2-hydroxy-2-(8-hydroxy-2-oxo-1H-quinolin-5-y1)ethyl]amino]ethyl-methyl-carbamoyl]phenyl]methyl]-7-azaspiro[3.5]nonan-2-y1]N-(2-phenylphenyl)carbamate；ditrifluoroacetic acid
Figure GPA0000234146600000981
[7- [ [4- [2- [ [ (2R) -2- [ tert-butyl (dimethyl) silyl ] oxy-2- (8-hydroxy-2-oxo-1H-quinolin-5-yl) ethyl ] amino ] ethylmethylcarbamoyl ] phenyl ] methyl ] -7-azaspiro [3.5] nonan-2-yl ] N- (2-phenoxyphenyl) carbamate (18E) (0.800g, 0.948mmol) was dissolved in dichloromethane (10m L), triethylamine trihydrofluoride salt (0.764g, 4.74mmol) was added, reaction overnight at room temperature, the reaction liquid was adjusted to alkali with saturated sodium bicarbonate, extraction was performed with 8% methanol/dichloromethane (v/v ═ 8: 92, 100m L×), the organic phase was washed with saturated aqueous sodium chloride solution (50m L× 1), anhydrous sodium sulfate was dried, the filtrate was concentrated under reduced pressure, the residue was separated and purified with a liquid-phase preparative column (conditions: C18 preparative column, mobile phase was 0.05% aqueous phase containing 0.05% of ethyl-2-phenyl-ethyl carbamate (0.7% ethyl-7% phenyl-7% of the title compound, eluent was obtained at a flow rate of ethyl-7-phenyl-7% ethyl-7% of ethyl-phenyl-7-amino-phenyl-7% of the title compound (TFA).
1H NMR(400MHz，CD3OD)8.43(d，1H)，7.64-7.57(m，5H)，7.48-7.27(m，9H)，7.06(d，1H)，6.66(d，1H)，5.47(s，1H)，4.36(s，2H)，3.94(s，2H)，3.53-3.35(m，6H)，3.04(m，5H)，2.46(s，1H)，2.25(s，1H)，1.97-1.87(m，7H).
LCMS m/z＝365.8[(M+2)/2]。
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/m L G418(sigma G5013) and 250. mu.g/m L Zeocin (Invitrogen ant-zn-5p) at 37 ℃ with 5% CO2Culturing under the condition to achieve 90-100% fusion. Washing with PBS/5mM EDTA to separate cells, centrifuging, collecting, resuspending and counting the cells in 0.1% BSA (BOVOGEN BSAS 100) phenol red-free Ham's F12 medium (Invitrogen 11039-6cells/m L. Add 15ml of cell suspension to a 50m L centrifuge tube, add Coelenterazine-h (promega S2011) to a final concentration of 5. mu.M, wrap with tinfoil to protect from light, incubate for 4 hours at 20 ℃ on a rotary shaker, dilute the cells with 0.1% BSA/phenol-free Red Ham' S F12 medium to a final concentration of 5.0 × 105cells/m L, cells were placed on a rotary shaker at low speed and incubated at room temperature for at least 1 hour the compounds of the examples were made up in DMSO as 10mM stock, 0.1% BSA/phenol red free Ham's F12 medium gradient diluted (log (M): 7, -8, -9, -10, -11), 96 well plates were added, 50L per well 50L cell suspension (25000 cells/well) was added per well and incubated at room temperature for 15 minutes the 96 well plates were placed in a microplate reader (Perkin Elmer, Envision) and labeled with microplate50L acetylcholine chloride (SigmaA6625) solution was added to each well of the instrument at 112.92nM (hM3), 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 results of the inhibitory activity of the test compounds on human muscarinic M3 receptor
Example numbering hM3 receptor IC50(nM)
Compound 1 2.97
Compound 3 3.25
Compound 5 0.44
Compound 6 0.70
Compound 7 0.75
Compound 8 1.34
Compound 9 0.60
Compound 10 1.63
Compound 11 5.75
Compound 16 2.04
Compound 17 1.39
And (4) conclusion: the compound has obvious inhibitory activity on human muscarinic M3 receptor.
Test example 2 agonistic Activity on human adrenergic β 2 receptor
Agonist activity of the compounds of the examples on human adrenergic receptors was determined by L ANCE Ultra cAMP Assay.
CHO cells (PerkinElmer, ES-034-CF) stably expressing human adrenergic receptor (h β 2) were cultured in MEM-alpha medium (Invitrogen 12561-056) containing 10% Fetal Bovine Serum (FBS) (Gibico 10099-141) and 250. mu.g/m L Zeocin (Invivogen ant-zn-5p) at 37 ℃ and 5% CO2Culturing under conditions to achieve 90-100% fusion and detecting the agonism of cAMP in the examples using L ANCE Ultra cAMP Assay kit (PerkinElmer TRF0263) cells were separated with PBS/5mM EDTA, harvested by centrifugation, resuspended in Stimulation Buffer (1 × HBSS, 5mM HEPES, 0.5mM IBMX, 0.1% BSA, pH7.4), and adjusted to 6 × 10 cell concentration5cells/ml. example compounds were made up in DMSO as 10mM stock, diluted in a Ststimulation Buffer gradient and added to a 384 well plate at 5. mu.l per well 5. mu. L cell suspension (3000 cells/well) was added per well, and after incubation for 30 minutes at room temperature, 5. mu.l 4x Eu-cAMP tracer working solution was added per well, followed by 5. mu.l 4x Ulight-anti-cAMP working solution per well and incubation for 1 hour at room temperature the 384 well plate was tested for TR-FRET using a microplate reader (Perkin Elmer, Envision), and EC was calculated and analyzed using origin7.550. Stimulation of human adrenergic receptors by the Compounds of the inventionKinetic Activity measured by the above experiment, measured EC50The values are shown in Table 2.
Table 2 results of testing the agonist activity of the compounds at the human adrenergic β 2 receptor
Example numbering h β 2 receptor EC50(nM)
Compound 1 2.8
Compound 2 8.4
Compound 3 7.75
Compound 4 1.25
Compound 6 1.02
Compound 7 3.0
Compound 9 3.4
Compound 10 2.0
Compound 11 0.90
Compound 12 1.08
Compound 13 0.85
Isomer 1 of compound 13 2.34
Isomer 2 of compound 13 2.63
Compound 16 4.90
Compound 17 1.80
Compound 18 1.96
The conclusion is that the compound of the invention has obvious activation activity on β 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 test compound was prepared into 0.6mM stock solution using 83% absolute ethanol + 17% Tween 80. It is diluted 500 times with water before 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 ExpTher 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 GPA0000234146600001011
And (4) conclusion: the compound has better inhibition effect on acetylcholine-induced bronchoconstriction of guinea pigs than positive control, and still has good bronchoconstriction inhibition effect after being administrated for 24 hours.

Claims (14)

1. A compound shown in a general formula (I) or a stereoisomer and a pharmaceutically acceptable salt thereof,
Figure FDA0002493146940000011
wherein:
R1is selected from
Figure FDA0002493146940000012
R1aSelected from H, C3-10Carbocyclyl or 3-to 8-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3Or C1-4Substituent of alkyl(iii) the heterocyclyl contains 1 to 3 heteroatoms selected from N, O or S;
R1bis selected from C3-10Carbocyclyl or 3-to 8-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3Or C1-4Alkyl, said heterocyclyl containing 1 to 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy or C1-6An alkyl group;
R1eand R1fEach independently selected from F, Cl, Br, I or CF3；
R2Selected from the group consisting of a bond and C1-6Alkylene optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I or C1-4Alkyl is substituted by a substituent;
l is selected from a bond or
Figure FDA0002493146940000013
And Y is directly connected to R2Connecting;
R3selected from the group consisting of a bond and C1-6Alkylene optionally further substituted with 0 to 5 substituents selected from R3aSubstituted with the substituent(s);
and R is2L and R3The three can not be simultaneously keys;
R4、R5each independently selected from H or C1-4An alkyl group;
R3aselected from F, Cl, Br, I or C1-4An alkyl group;
A1and A4Each independently selected from C6-10Arylene, wherein said arylene is optionally further substituted with 0 to 5 substituents selected from R6Substituted with the substituent(s);
A2and A3Each independently selected from C1-6Alkylene optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I or C1-4Alkyl is substituted by a substituent;
y and X are each independently selected fromBond, -C (═ O) NR7-、-NR7C(＝O)-、-OC(＝O)NR7-or-NR7C(＝O)O-；
R6Selected from F, Cl, Br, I, OH, C1-4Alkyl or C1-4An alkoxy group;
R7each independently selected from H or C1-4An alkyl group;
a is 0 or 1;
b is 0 or 1;
m, n, p or q are each independently selected from 0 or 1;
b is selected from
Figure FDA0002493146940000021
Wherein Q is selected from-CH ═ CH-, -O-, -S-, -CH2O-or-OCH2-。
2. A compound according to claim 1, or a stereoisomer or pharmaceutically acceptable salt thereof,
R1aselected from H, C3-7Carbocyclyl or 3-to 6-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CF3OH or C1-4Alkyl, said heterocyclyl containing 1 to 3 heteroatoms selected from N, O or S;
R1bis selected from C3-7Carbocyclyl or 3-to 6-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CF3OH or C1-4Alkyl, said heterocyclyl containing 1 to 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy or C1-4An alkyl group;
R1eand R1fEach independently selected from F, Cl, Br, I or CF3；
R6Each independently selected from F, Cl, Br, I, OH and C1-4Alkyl or C1-4An alkoxy group.
3. A compound according to claim 2, or a stereoisomer or pharmaceutically acceptable salt thereof,
R1aselected from H, phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, thienyl, furyl or pyridyl, said phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, thienyl, furyl or pyridyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, CF3OH or C1-4Alkyl is substituted by a substituent;
R1bselected from phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, thienyl, furyl or pyridyl, said phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, thienyl, furyl or pyridyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, CF3OH or C1-4Alkyl is substituted by a substituent;
R1cselected from H, hydroxy, methyl or ethyl;
R1eand R1fEach independently selected from F, Cl, Br, I or CF3；
R2Selected from the group consisting of a bond, methylene, ethylene, propylene, butylene or pentylene, said methylene, ethylene, propylene, butylene or pentylene being optionally further substituted with 0 to 5 substituents selected from the group consisting of F, Cl, Br, I, methyl or ethyl;
R3selected from a bond, methylene, ethylene, propylene, butylene, pentylene or
Figure FDA0002493146940000031
The methylene, ethylene, propylene, butylene, pentylene or
Figure FDA0002493146940000032
Optionally further substituted with 0 to 5 substituents selected from F, Cl, Br, I, cyano, OH, methyl, ethyl, methoxy or ethoxy;
R4、R5each independently selected from H, methyl or ethyl;
b is selected from
Figure FDA0002493146940000033
Figure FDA0002493146940000034
A1And A4Each independently selected from phenylene, thienylene, furanylene, or pyridinylene, said phenylene, thienylene, furanylene, or pyridinylene optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, methyl, ethyl, propyl, isopropyl, methoxy, or ethoxy;
A2and A3Each independently selected from methylene, ethylene, propylene, butylene or pentylene;
y and X are each independently selected from the group consisting of a bond, -C (═ O) NR7-、-NR7C(＝O)-、-OC(＝O)NR7-or-NR7C(＝O)O-；
R7Selected from H, methyl, ethyl or propyl;
a is 0 or 1;
b is 0 or 1.
4. A compound according to claim 3, or a stereoisomer or pharmaceutically acceptable salt thereof,
R1is selected from
Figure FDA0002493146940000041
Figure FDA0002493146940000042
R2Selected from the group consisting of a bond, methylene, ethylene, propylene, -CH2CH(CH3)-、-CH2C(CH3)2-、-C(CH3)2CH2-、-CH(CH3)CH2-, butylene, -CH (CH)3)CH2CH2-、-CH2CH(CH3)CH2-、-CH2CH(CH3)CH2-or pentylene;
R3selected from the group consisting of a bond, methylene, ethylene, propylene, -CH2CH(CH3)-、-CH2C(CH3)2-、-C(CH3)2CH2-、-CH(CH3)CH2-, butylene, -CH (CH)3)CH2CH2-、-CH2CH(CH3)CH2-、-CH2CH(CH3)CH2-、
Figure FDA0002493146940000043
Or a pentylene group;
A1and A4Each independently selected from phenylene optionally further substituted with 0 to 4 substituents optionally selected from F, Cl, Br, methyl, ethyl, methoxy or ethoxy.
5. A compound as shown below or a stereoisomer or pharmaceutically acceptable salt thereof:
Figure FDA0002493146940000044
Figure FDA0002493146940000051
6. a pharmaceutical composition comprising a therapeutically effective dose of a compound of any one of claims 1-5, or a stereoisomer or 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.
7. The pharmaceutical composition of claim 6, wherein the additional therapeutic agent is selected from one or more of a PDE4 inhibitor, an M receptor antagonist, a corticosteroid, and a β -adrenoceptor agonist.
8. Use of a compound according to any one of claims 1 to 5, or a stereoisomer or pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of an obstructive airways disease.
9. Use of a compound according to any one of claims 1 to 5, or a stereoisomer or pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of asthma, chronic obstructive pulmonary disease or bronchitis.
10. An intermediate selected from a compound of formula (II) or a stereoisomer thereof:
Figure FDA0002493146940000061
wherein:
R1is selected from
Figure FDA0002493146940000062
R1aSelected from H, C3-10Carbocyclyl or 3-to 8-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3Or C1-4Alkyl, said heterocyclyl containing 1 to 3 heteroatoms selected from N, O or S;
R1bis selected from C3-10Carbocyclyl or 3-to 8-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3Or C1-4Alkyl, said heterocyclyl containing 1 to 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy or C1-6An alkyl group;
R1eand R1fEach independently selected from F, Cl, Br, I or CF3；
R9Selected from H or an amino protecting group;
a is 0 or 1;
b is 0 or 1.
11. The intermediate of claim 10, wherein:
R1is selected from
Figure FDA0002493146940000071
R9Selected from H, benzyl, p-methoxybenzyl, tert-butoxycarbonyl, benzyloxycarbonyl, acetyl or benzoyl.
12. An intermediate selected from one of the following compounds:
Figure FDA0002493146940000072
13. an intermediate selected from a compound of formula (III) or a stereoisomer thereof:
Figure FDA0002493146940000073
R1is selected from
Figure FDA0002493146940000074
R1aSelected from H, C3-10Carbocyclyl or 3-to 8-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3Or C1-4Alkyl, said heterocyclyl containing 1 to 3 substituents selected fromA heteroatom from N, O or S;
R1bis selected from C3-10Carbocyclyl or 3-to 8-membered heterocyclyl, said carbocyclyl or heterocyclyl being optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, CH2F、CHF2、CF3、OH、OCH2F、OCHF2、OCF3Or C1-4Alkyl, said heterocyclyl containing 1 to 3 heteroatoms selected from N, O or S;
R1cselected from H, hydroxy or C1-6An alkyl group;
R1eand R1fEach independently selected from F, Cl, Br, I or CF3；
R2Selected from the group consisting of a bond and C1-6Alkylene optionally further substituted by 0 to 5 substituents selected from F, Cl, Br, I or C1-4Alkyl is substituted by a substituent;
w is selected from OH, carboxyl, phenyl, -C (═ O) NRx-phenyl, -C (═ O) NRx-C1-4alkylene-C (═ O) OH, -C (═ O) NRx-C1-4alkylene-C (═ O) OC1-4Alkyl, -C (═ O) NRx-C1-4alkylene-C (═ O) NRx-phenyl, -C (═ O) NRx-phenylene-C (═ O) NRx-phenyl group,
Figure FDA0002493146940000081
Figure FDA0002493146940000082
Said phenyl group is optionally further substituted with 0 to 4 substituents selected from F, Cl, Br, I, methoxy, ethoxy,
Figure FDA0002493146940000083
Formyl, carboxy, -C (═ O) OC1-4Alkyl, aryl, heteroaryl, and heteroaryl,
Figure FDA0002493146940000084
Figure FDA0002493146940000085
Substituted with the substituent(s);
Rxis selected from H or C1-4An alkyl group;
R3selected from the group consisting of a bond and C1-6Alkylene optionally further substituted with 0 to 5 substituents selected from R3aIs substituted by a substituent of (A), R3aSelected from F, Cl, Br, I or C1-4An alkyl group;
b is selected from
Figure FDA0002493146940000086
Wherein Q is selected from-CH ═ CH-, -O-, -S-, -CH2O-or-OCH2-；
a is 0 or 1;
b is 0 or 1.
14. An intermediate selected from one of the following compounds:
Figure FDA0002493146940000087
Figure FDA0002493146940000091
Figure FDA0002493146940000101
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