CN114621243A

CN114621243A - Sulfonamide derivative and application thereof

Info

Publication number: CN114621243A
Application number: CN202111529297.7A
Authority: CN
Inventors: 李云飞; 莫明广; 杨煜文; 张芳
Original assignee: Shanghai Tuojie Biomedical Technology Co ltd
Current assignee: Shanghai Tuojie Biomedical Technology Co ltd
Priority date: 2020-12-14
Filing date: 2021-12-14
Publication date: 2022-06-14
Anticipated expiration: 2041-12-14
Also published as: CN114621243B

Abstract

The disclosure relates to sulfonamide derivatives and uses thereof. Specifically, the present disclosure provides a compound represented by formula I or a pharmaceutically acceptable salt thereof, wherein ring a and B, X¹～X⁴、R¹～R⁸As defined herein.

Description

Sulfonamide derivative and application thereof

Technical Field

The disclosure belongs to the field of medicines, and relates to a sulfonamide derivative and application thereof.

Background

Semicarbazide-sensitive amine oxidases (SSAO) are a class of dopamine quinone-containing amine oxidases that are members of the semicarbazide-sensitive amine oxidase family, also known as vascular adhesion protein-1, VAP-1(vascular adhesion protein 1). Numerous studies have demonstrated that SSAO and its metabolites are closely related to inflammatory-related diseases such as atherosclerosis, diabetes and its complications, obesity, stroke, chronic kidney disease, retinopathy, Chronic Obstructive Pulmonary Disease (COPD), autoimmune diseases, multiple sclerosis, rheumatoid arthritis, alzheimer's disease, and the like.

Several known MAO inhibitors, such as mofetil, have been synthesized and studies have shown that mofetil inhibits experimental autoimmune encephalomyelitis (US20060025438),

WO2009066152 describes 3-substituted 3-haloalkallylamine SSAO/VAP-1 inhibitors and claims them as a treatment for inflammatory diseases,

WO2013163675 develops a new class of 3-haloalkallylamine SSAO/VAP-1 inhibitors on the basis of the above, and the following compounds are exemplified:

in addition, other 3-haloalkallylamine-based SSAO/VAP-1 inhibitors have been reported in succession, such as CN109251166, CN109810041, CN110938059, CN108778278, CN109988093, CN109988106, CN109988109, WO2018027892, WO2018149226, WO2020233583, WO2007120528, WO2018196677, WO 202006383854, WO2020089025, WO2020089026, WO2020125776, and the like, however, no SSAO/VAP-1 inhibitor is currently marketed, and the disclosed compounds are not disclosed in any literature and exhibit specific VAP-1 inhibitory effects.

Disclosure of Invention

The disclosure provides a compound of formula I or a pharmaceutically acceptable salt thereof

Wherein R is¹And R²Independently selected from hydrogen, deuterium, chlorine, fluorine;

R³and R⁴Independently selected from hydrogen, deuterium, C_1-6An alkyl group optionally substituted with one or more R^A1Each independently substituted, R^A1Selected from halogen (e.g., fluorine, chlorine), deuterium, hydroxy, nitro, cyano or amino;

R⁵and R⁶Independently selected from hydrogen, deuterium, C_1-6An alkyl group optionally substituted with one or more R^A2Each independently substituted, R^A2Selected from halogen (e.g., fluorine, chlorine), deuterium, hydroxy, nitro, cyano or amino;

R⁷and R⁸Independently selected from hydrogen, deuterium, C_1-6An alkyl group optionally substituted with one or more R^A3Each independently substituted, R^A3Selected from halogen (e.g., fluorine, chlorine), deuterium, hydroxy, nitro, cyano or amino;

ring A is selected from a 4-to 6-membered heterocyclic ring, said Ring A being optionally substituted with one or more R^A4Substituted; r^A4Each independently selected from halogen (e.g. fluoro, chloro), deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -SR^1a、-S(O)R^1a、-S(O)₂R^1a、-NR^1a(R^1b)、-(CH₂)_oCOR^1a、-(CH₂)_oNHCOR^1a、-(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCOR^1a；

R^1aOr R^1bEach independently selected from hydrogen, deuterium, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl optionally substituted with one or more groups selected from halo (e.g., fluoro, chloro), deuterium, hydroxy, oxo, nitro, cyano or amino;

ring B is selected from a 3-to 6-membered carbocyclic ring, a 3-to 6-membered heterocyclic ring, or a 5-to 6-membered heteroaromatic ringA ring, said ring B being optionally substituted with one or more R^A5Substituted; r^A5Selected from halogen (e.g. fluorine, chlorine), deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl, heteroaryl, -SR^2a、-S(O)R^2a、-S(O)₂R^2a、-NR^2a(R^2b)、-(CH₂)_pCOR^2a、-(CH₂)_pNHCOR^2a、-(CH₂)_pCONR^2a(R^2b)、-(CH₂)_pOCONR^2a(R^2b)、-N(CH₂)_pCONR^2a(R^2b)、-N(CH₂)_pCOR^2a；

R^2aOr R^2bEach independently selected from hydrogen, deuterium, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl being optionally substituted by one or more groups selected from halogen (e.g. fluoro, chloro), deuterium, hydroxy, oxo, nitro, cyano or amino;

and ring a is connected to ring B in a fused or spiro ring form;

X¹、X²、X³and X⁴Each independently is-CH-or-N-, and is not simultaneously-N-;

o and p are each selected from integers between 0 and 3.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof¹Selected from hydrogen, R²Selected from fluorine.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof¹Selected from fluorine, R²Selected from hydrogen.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof¹Selected from hydrogen, R²Selected from chlorine.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof¹Selected from chlorine, R²Selected from hydrogen.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof⁷And R⁸Independently selected from C_1-6An alkyl group optionally substituted with one or more R^A3Substituted, R^A3Each independently selected from halogen, deuterium, hydroxy, nitro, cyano or amino.

In other embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof⁷And R⁸Independently selected from methyl, ethyl or propyl, further optionally substituted with one or more R^A3Substituted, R^A3Each independently selected from halogen, deuterium, hydroxy, nitro, cyano or amino.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof⁵And R⁶Independently selected from hydrogen or deuterium.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof⁵、R⁶、R⁷And R⁸Selected from hydrogen.

In another aspect, some embodiments provide that ring A is selected from the group consisting of

Further, ring A is optionally substituted with 1 to 3R^A4Substituted, R^A4The foregoing definitions.

In other embodiments, there is provided a compound of formula I or a pharmaceutically acceptable salt thereof

Is selected from

Wherein n and m are each independently selected from integers between 0 and 4, the sum of n and m is not more than 4, and Y is selected from-NH-, O-CH₂-or-S-, further

By 1 to 3R^A4Or R^A5Substituted, R^A4、R^A5As defined above. In some embodiments, wherein Y is selected from-NH-, O, -CH₂-; the sum of n and m is 1,2, 3 or 4.

In some embodiments, ring B is selected from 3 to 6 carbocycles, preferably 3 to 6 carbocycles, in a compound of formula I or a pharmaceutically acceptable salt thereof

Further, ring B is optionally substituted with 1 to 3R^A5Substituted, R^A5As defined above.

In other embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof

Is selected from

Further, the method can be used for preparing a novel liquid crystal display

By 1 to 3R^A4Or R^A5Substituted, R^A4、R^A5As defined above.

Is selected from

Further, the method can be used for preparing a novel material

By 1 to 3R^A4Or R^A5Substituted, R^A4、R^A5As defined above.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A4Each independently selected from halogen, deuterium, oxo (═ O), hydroxy, cyano or amino.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A4Each independently selected from C_1-6Alkyl or C_1-6An alkoxy group.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A4Each independently selected from C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl or heteroaryl.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A4Each independently selected from-NR^1a(R^1b)、-(CH₂)_oCOR^1a、-(CH₂)_oNHCOR^1a、-(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCOR^1a，o、R^1aOr R^1bAs defined above. In some embodiments, o is 0, 1, or 2 in the compound of formula I or a pharmaceutically acceptable salt thereof.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^1aOr R^1bSelected from hydrogen, deuterium, hydroxy, C_1-6Alkyl or C_1-6Alkoxy, said alkyl or alkoxy being optionally substituted with 1 to 3 substituents selected from halogen. In other embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^1aOr R^1bSelected from hydrogen, deuterium, hydroxyl, methyl, ethyl, methoxy or ethoxy.

On the other hand, some embodimentsR in the compound shown in the formula I or the pharmaceutically acceptable salt thereof^A5Each independently selected from halogen, deuterium, oxo (═ O), hydroxy, cyano or amino.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A5Each independently selected from oxo (═ O), hydroxy, halogen, C_1-6Alkyl or C_1-6An alkoxy group.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A5Each independently selected from C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl or heteroaryl.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A5Each independently selected from oxo (═ O), hydroxy, C_1-6Alkyl, -NR^2a(R^2b)、-(CH₂)_pCOR^2a、-(CH₂)_pNHCOR^2a、-(CH₂)_pCONR^2a(R^2b)、-(CH₂)_pCONR^2a(R^2b)、-N(CH₂)_pCONR^2a(R^2b)、-N(CH₂)_pCOR^2a，p、R^2aOr R^2bAs defined above.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A5Each independently selected from- (CH)₂)_pCOR^2aOr- (CH)₂)_pCONR^2a(R^2b). In some embodiments, p is 0, 1, or 2 in a compound of formula I or a pharmaceutically acceptable salt thereof.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A5Each independently selected from oxo (═ O), hydroxy, C_1-6Alkyl, -NR^2a(R^2b)、-COR^2a、-CH₂NHCOR^2a、-CONR^2a(R^2b)、-CH₂CONR^2a(R^2b)、-N(CH₂)CONR^2a(R^2b)、-NCH₂COR^2a、-NHCOR^2a，R^2aOr R^2bAs defined above.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A5Each independently selected from oxo (═ O), hydroxy, C_1-6Alkyl, -COR^2a、-CONR^2a(R^2b)、-NHCOR^2a，R^2aOr R^2bAs defined above.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^A5Selected from fluorine, oxo, C_1-3Alkyl, -COOH, -COC_1-3Alkoxy, -CONHC_1-3An alkoxy group.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^2aOr R^2bSelected from hydrogen, deuterium, hydroxy, C_1-6Alkyl or C_1-6Alkoxy, said alkyl, alkoxy being optionally substituted with 1 to 3 substituents selected from halogen. In other embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^2aOr R^2bSelected from hydrogen, deuterium, hydroxy, methyl, ethyl, methoxy or ethoxy.

In another aspect, in some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^1aOr R^1bIs selected from C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, said cycloalkyl or heterocycloalkyl optionally substituted with 1 to 3 substituents selected from halogen, deuterium, hydroxy, oxo, nitro, cyano or amino.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof^2aOr R^2bIs selected from C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, said cycloalkyl or heterocycloalkyl optionally substituted with 1 to 3 substituents selected from halogen, deuterium, hydroxy, oxo, nitro, cyano or amino.

In another aspect, the compound of formula I or a pharmaceutically acceptable salt thereof has ring B and ring A, wherein ring B is selected from a 5-membered heteroaromatic ring. In some embodiments, ring B is selected from the group consisting of

Further, ring B is optionally substituted with 1 to 3R^A5Substituted, R^A5As defined above. In other embodiments, there is provided a compound of formula I or a pharmaceutically acceptable salt thereof

Is selected from

Which is further substituted by 1 to 3R^A4Or R^A5And (4) substitution.

In some embodiments, ring B is selected from the group consisting of

Further, ring B is optionally substituted with 1 to 3R^A5Substituted, R^A5As defined above. In other embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof

Is selected from

Which is further substituted by 1 to 3R^A4Or R^A5Substituted, R^A5As defined above.

In other embodiments, ring B is selected from the group consisting of

Is selected from

Further, the method can be used for preparing a novel material

By 1 to 3R^A4Or R^A5Substituted, R^A4、R^A5As defined above. In other embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof

By 1 to 3R^A5Substituted, R^A5Each independently oxo (═ O) or C_1-6Alkyl (e.g., methyl, ethyl, or propyl).

In another aspect, in some embodiments, X in the compound of formula I or a pharmaceutically acceptable salt thereof²Is selected from-N-, X¹、X³、X⁴Is selected from-CH-; or X³Is selected from-N-, X¹、X²、X⁴Is selected from-CH-,

in some embodiments, X in the compound of formula I or a pharmaceutically acceptable salt thereof¹、X²、X³、X⁴Is selected from-CH-,

in some embodiments, X in the compound of formula I or a pharmaceutically acceptable salt thereof¹、X⁴Is selected from-N-, X²、X³Is selected from-CH-,

in some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof

Is selected from

Further, the method can be used for preparing a novel liquid crystal display

By 1 to 2R^A4Or R^A5And (4) substituting.

Is selected from

Further, the method can be used for preparing a novel material

Is 1 to 2 of R^A4Or R^A5And (4) substituting.

In other embodiments, there is provided a compound of formula I or a pharmaceutically acceptable salt thereof as

In some embodiments, the compound of formula I, or a pharmaceutically acceptable salt thereof, is

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof³Selected from hydrogen or C_1-6Alkyl (including but not limited to methyl, ethyl or propyl).

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof³Is selected fromC_1-6Alkyl (including but not limited to methyl, ethyl or propyl), optionally substituted with one or more R^A1Substituted, R^A1Selected from halogen, deuterium, hydroxyl, nitro, cyano or amino.

Further, in some embodiments, R^A5Is selected from- (CH)₂)_pCOR^2aWherein R is^2aIs selected from C_1-6Alkyl optionally substituted with one or more groups selected from halo, deuterium, hydroxy, oxo, nitro, cyano or amino, p ═ 0, 1 or 2.

In some embodiments, R^A5Is selected from- (CH)₂)_pCONR^2a(R^2b) Wherein R is^1aSelected from hydrogen, R^2aIs selected from C_1-6Alkyl optionally substituted with one or more groups selected from halo, deuterium, hydroxy, oxo, nitro, cyano or amino, p ═ 0, 1 or 2. Further, in some embodiments, R^A4Is selected from- (CH)₂)_pCOR^2aWherein R is^2aIs selected from C_1-6Alkyl optionally substituted with one or more groups selected from halo, deuterium, hydroxy, oxo, nitro, cyano or amino, p ═ 0, 1 or 2.

In some embodiments, R^A4Is selected from- (CH)₂)_pCONR^2a(R^2b) Wherein R is^1aSelected from hydrogen, R^2aIs selected from C_1-6Alkyl optionally substituted with one or more groups selected from halo, deuterium, hydroxy, oxo, nitro, cyano or amino, p ═ 0, 1 or 2.

In some embodiments, R in the compound of formula I or a pharmaceutically acceptable salt thereof³Selected from hydrogen, methyl or ethyl.

Typical compounds of formula I, or pharmaceutically acceptable salts thereof, include, but are not limited to:

wherein

Including the E or Z configuration.

In some embodiments, a compound of formula I, or a pharmaceutically acceptable salt thereof, comprises:

the present disclosure also provides a pharmaceutical composition comprising at least one therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

In some embodiments, the unit dose of the pharmaceutical composition is from 0.001mg to 1000 mg.

In certain embodiments, the pharmaceutical composition comprises 0.01 to 99.99% of the aforementioned compound or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition comprises 0.1-99.9% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition comprises from 0.5% to 99.5% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition comprises 1% to 99% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition comprises 2% to 98% of the aforementioned compound or a pharmaceutically acceptable salt thereof.

In certain embodiments, the pharmaceutical composition comprises from 0.01% to 99.99% of a pharmaceutically acceptable excipient, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1% to 99.9% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition contains 0.5% to 99.5% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition contains 1% to 99% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition contains 2% to 98% of a pharmaceutically acceptable excipient.

The present disclosure also provides a method of preventing and/or treating a patient suffering from a condition associated with SSAO or SSAO/VAP-1 by administering to said patient a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof, as described above, or a pharmaceutical composition as described above.

In some embodiments, the SSAO or SSAO/VAP-1 related disorder is selected from inflammation, diabetes, an ocular disease, fibrosis, a neuroinflammatory disease, or cancer.

The present disclosure also provides a method for preventing and/or treating a patient suffering from inflammation, diabetes, ocular diseases, fibrosis, neuroinflammatory diseases, or cancer, comprising administering to the patient a therapeutically effective amount of a compound of formula I as set forth above or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as set forth above.

The present disclosure also provides the use of a compound of formula I as described above or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of the foregoing for the manufacture of a medicament for the prevention and/or treatment of a condition associated with SSAO or SSAO/VAP-1. In some embodiments, the PDE-related disorder is preferably inflammation, diabetes, an ocular disease, fibrosis, a neuroinflammatory disease, or cancer.

The present disclosure also provides a use of the compound represented by the formula I or a pharmaceutically acceptable salt thereof or the pharmaceutical composition for the preparation of a medicament for preventing and/or treating inflammation, diabetes, an eye disease, fibrosis, a neuroinflammatory disease or cancer.

The pharmaceutically acceptable salts of the compounds described in this disclosure may be selected from inorganic or organic salts.

The disclosed compounds may exist in specific geometric or stereoisomeric forms. The present disclosure contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which fall within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present disclosure. The compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from a racemic mixture or synthesized by using chiral starting materials or chiral reagents.

Optically active (R) -and (S) -isomers as well as D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one of the enantiomers of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), diastereomeric salts are formed with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art, and the pure enantiomers are recovered. Furthermore, separation of enantiomers and diastereomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amines).

In the chemical structure of the compounds described in the present disclosure, a bond

Denotes an unspecified configuration, i.e. a bond if a chiral isomer is present in the chemical structure

Can be that

Or at the same time contain

Two configurations. Key with a key body

Indicates unspecified configurations, including cis (E) or trans (Z) configurations. Or as described in the present disclosure

Refers to a double bond, and the structure bonded by the bond can be "cis isomer" or "trans isomer" or "a mixture of cis and trans isomers in any ratio", for example formula E represents E-1, formula E-2, or a mixture of both in any ratio:

the compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also referred to as proton transfer tautomers) include interconversion via proton migration, such as keto-enol and imine-enamine, lactam-lactam isomerizations. An example of a lactam-lactam equilibrium is between A and B as shown below.

All compounds in this disclosure can be drawn as form a or form B. All tautomeric forms are within the scope of the disclosure. The naming of the compounds does not exclude any tautomers.

The disclosure also includes some isotopically-labeled compounds of the present disclosure that are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as respectively²H、³H、¹¹C、¹³C、¹⁴C、¹³N、¹⁵N、¹⁵O、¹⁷O、¹⁸O、³¹P、³²P、³⁵S、¹⁸F、¹²³I、¹²⁵I and³⁶cl, and the like.

Unless otherwise stated, when a position is specifically designated as deuterium (D), that position is understood to be deuterium having an abundance that is at least 1000 times greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 10% deuterium incorporation). The compound of examples can have a natural abundance of deuterium greater than at least 1000 times the abundance of deuterium, at least 2000 times the abundance of deuterium, at least 3000 times the abundance of deuterium, at least 4000 times the abundance of deuterium, at least 5000 times the abundance of deuterium, at least 6000 times the abundance of deuterium, or more abundant deuterium. The disclosure also includes various deuterated forms of the compounds of formula (I). Each available hydrogen atom attached to a carbon atom may be independently replaced by a deuterium atom. The person skilled in the art is able to synthesize the deuterated forms of the compounds of the formula (I) with reference to the relevant literature. Commercially available deuterated starting materials can be used in preparing the deuterated forms of the compounds of formula (I), or they can be synthesized using conventional techniques using deuterated reagents including, but not limited to, deuterated boranes, trideuteroborane tetrahydrofuran solutions, deuterated lithium aluminum hydrides, deuterated iodoethanes, deuterated iodomethanes, and the like.

"optionally" or "optionally" means that the subsequently described event or circumstance can, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. For example "C optionally substituted by halogen or cyano_1-6Alkyl "means that halogen or cyano may, but need not, be present, and that the description includes alkyl substituted with halogen orA cyano group and a alkyl group which is not substituted by halogen or cyano.

Interpretation of terms:

"pharmaceutical composition" means a mixture containing one or more compounds described herein, or a physiologically acceptable salt or prodrug thereof, in admixture with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate absorption of the active ingredient and exert biological activity.

"pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that has been approved by the U.S. food and drug administration for use in humans or livestock animals.

An "effective amount" or "therapeutically effective amount" as referred to in this disclosure includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical condition. An effective amount also means an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on the following factors: such as the condition to be treated, the general health of the patient, the method and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

"alkyl" refers to a saturated aliphatic hydrocarbon group, including straight and branched chain groups of 1 to 20 carbon atoms. An alkyl group having 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, and various branched isomers thereof, and the like. Alkyl groups may be substituted or unsubstituted, and when substituted, substituents may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen, deuterium, hydroxy, nitro, cyano or amino.

The term "cycloalkyl" or "carbocycle"refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing from 3 to 20 carbon atoms, preferably from 3 to 7 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups. Cycloalkyl groups may be substituted or unsubstituted, and when substituted, the substituents may be substituted at any available point of attachment, preferably one or more groups independently selected from halogen, deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl, heteroaryl, -SR ', -S (O) R', -S (O)₂R’、-NR’(R”)、-(CH₂)_pCOR’、-(CH₂)_pNHCOR’、-(CH₂)_pCONR’(R”)、-(CH₂)_pOCONR’(R”)、-N(CH₂)_pCONR’(R”)、-N(CH₂)_pAnd (3) COR'. The cycloalkyl ring may be fused to an aryl or heteroaryl ring, wherein the ring to which the parent structure is attached is cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, and the like. Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl, heteroaryl, -SR ', -S (O) R', -S (O)₂R’、-NR’(R”)、-(CH₂)_pCOR’、-(CH₂)_pNHCOR’、-(CH₂)_pCONR’(R”)、-(CH₂)_pOCONR’(R”)、-N(CH₂)_pCONR’(R”)、-N(CH₂)_pCOR’。

The term "heterocycloalkyl" or "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 6 ring atoms, non-limiting example packages of "heterocycloalkyl" includeComprises the following steps:

and so on.

Heterocycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl, heteroaryl, -SR ', -S (O) R', -S (O)₂R’、-NR’(R”)、-(CH₂)_pCOR’、-(CH₂)_pNHCOR’、-(CH₂)_pCONR’(R”)、-(CH₂)_pOCONR’(R”)、-N(CH₂)_pCONR’(R”)、-N(CH₂)_pCOR’。

The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5-or 6-membered. For example. Non-limiting examples thereof include:

and so on.

Heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl, heteroaryl, -SR ', -S (O) R', -S (O)₂R’、-NR’(R”)、-(CH₂)_pCOR’、-(CH₂)_pNHCOR’、-(CH₂)_pCONR’(R”)、-(CH₂)_pOCONR’(R”)、-N(CH₂)_pCONR’(R”)、-N(CH₂)_pCOR’。

The disclosure of the inventionWherein R 'or R' is selected from hydrogen, deuterium, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl being optionally substituted with one or more groups selected from halogen, deuterium, hydroxy, oxo, nitro, cyano or amino.

The term "alkoxy" refers to-O- (alkyl) and-O- (unsubstituted cycloalkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from halogen, deuterium, hydroxy, oxo, nitro, cyano or amino.

The term "hydroxy" refers to an-OH group.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "cyano" refers to — CN.

The term "cyano" refers to-NH₂。

The term "nitro" means-NO₂。

The term "oxo" refers to the ═ O substituent.

"substituted" means that one or more, preferably up to 5, more preferably 1 to 3, hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (experimentally or theoretically) possible or impossible substitutions without undue effort.

Detailed Description

The present disclosure is further described below with reference to examples, but these examples do not limit the scope of the present disclosure.

Experimental procedures, in which specific conditions are not noted in the examples of the present disclosure, are generally performed under conventional conditions, or under conditions recommended by manufacturers of raw materials or commercial products. Reagents of specific sources are not indicated, and conventional reagents are purchased in the market.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS). NMR shift (. delta.) of 10^-6The units in (ppm) are given. NMR was measured using a Bruker AVANCE-400 NMR spectrometer using deuterated dimethyl sulfoxide (DMSO-d)₆) Deuterated chloroform (CDCl)₃) Deuterated Methanol (Methanol-d)₄) Internal standard is Tetramethylsilane (TMS).

The HPLC measurements were performed using an Agilent1100 HPLC, GAS15B DAD UV detector, Water Vbridge C18150 x 4.6mm 5um column.

The MS was measured using an Agilent6120 triple quadrupole mass spectrometer, G1315D DAD detector, Waters Xbridge C184.6 x 50mm,5um chromatography column, scanning in positive/negative ion mode with a mass scan range of 80-1200.

The silica gel plate for thin layer chromatography is HSGF254 silica gel plate of cigarette platform yellow sea, and the silica gel plate for Thin Layer Chromatography (TLC) is 0.2mm + -0.03 mm, and the specification of the product for thin layer chromatography separation and purification is 0.4mm-0.5 mm.

The flash column purification system used either Combiflash Rf150(TELEDYNE ISCO) or Isolara one (Biotage).

The forward column chromatography generally uses 200-300 mesh or 300-400 mesh silica gel of the Titan yellow sea as a carrier, or uses a hyperpure normal phase silica gel column (40-63 μm, 60g, 24g, 40g, 120g or other specifications) pre-filled by Santai in Changzhou.

Known starting materials in this disclosure can be synthesized by or according to methods known in the art, or can be purchased from companies such as Shanghai Tantan science, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoshi Chemical technology (Accela ChemBio Inc), Biddy medicine, and the like.

In the examples, the reactions were all carried out under a nitrogen atmosphere without specific indication.

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 connected with a hydrogen balloon with a volume of about 1L.

The hydrogen was produced by QPH-1L model hydrogen generator, Shanghai Quanpu scientific instruments.

The nitrogen atmosphere or the hydrogenation atmosphere is usually vacuumized, and filled with nitrogen or hydrogen, and the operation is repeated for 3 times.

In the examples, the solution means an aqueous solution unless otherwise specified.

In the examples, the reaction temperature is, unless otherwise specified, from 20 ℃ to 30 ℃ at room temperature.

The monitoring of the reaction progress in the examples employs Thin Layer Chromatography (TLC), a developing agent used for the reaction, an eluent system for column chromatography used for purifying a compound, and a developing agent system for thin layer chromatography, and the volume ratio of a solvent is adjusted according to the polarity of the compound, and may also be adjusted by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

Example 1

Synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-oxa-7-azaspiro [3.5] nonane (1)

Step 1: synthesis of 7- [ 6-chloropyridin-3-yl) sulfonyl ] -2-oxa-7-azaspiro [3.5] nonane (1c)

Compound 1b (563mg,2.59mmol) was dissolved in dichloromethane (10mL) and triethylamine (716mg,7.07mmol) and the compound 6-chloropyridine-3-sulfonyl chloride (1a) (0.50g,2.36mmol) were added and the reaction was monitored for completion by LC-MS at room temperature. Filtering, and separating with flash chromatography

Compound 1c (0.10g, yield 14%) was isolated.

MS(ESI)m/z:303.3[M+H]⁺

¹H NMR(400MHz,CDCl₃)δ8.76(d,J＝2.3Hz,1H),7.99(dd,J＝2.6,8.3Hz,1H),7.53(d,J＝8.0Hz,1H),4.37(s,4H),3.07-2.98(m,4H),2.03-1.97(m,4H).

Step 2: synthesis of 7- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-oxa-7-azaspiro [3.5] nonane (1d)

Compound 1c (59mg,0.20mmol) was dissolved in N-methylpyrrolidone (1mL) and triethylamine (30mg,0.29mmol) was added, followed by cooling to 0 ℃ with an ice-water bath. A solution of BB-1(40mg,0.20mmol) in tetrahydrofuran (2mL) and a solution of sodium tert-butoxide (28mg,0.29mmol) in dimethyl sulfoxide (0.5mL) were added. The reaction was allowed to proceed to LC-MS at room temperature to monitor completion of the reaction, water (10mL) was added, extraction was performed with ethyl acetate (10 mL. times.2), washing with saturated brine, drying over anhydrous magnesium sulfate, filtration and evaporation of the filtrate to dryness to give crude product 1d (50mg, yield 54%).

MS(ESI)m/z:472.2[M+H]⁺。

Wherein BB-1 is prepared by taking commercial BB-1a as a raw material:

dissolving compound BB-1a (6.62g,21.80mmol) in tetrahydrofuran (300mL), adding compound BB-1b (10.0g,21.60mmol), cooling to-60 deg.C, slowly adding NaHMDS (32.7mL,32.7mmol,1.0M), reacting at-50 deg.C to-60 deg.C for 1 hr, adding saturated aqueous ammonium chloride (500mL), extracting with ethyl acetate (500 mL. times.2), washing with saturated saline, drying with anhydrous magnesium sulfate, and separating with flash chromatography

BB-1c (1.50g, yield 21%) was isolated.

MS(ESI):m/z 220.2[M+H]⁺。¹H NMR(400MHz,CDCl₃)δ6.78-6.16(m,1H),4.34-3.96(m,2H),3.85-3.44(m,2H),1.34(d,J＝2.1Hz,9H),0.83-0.80(m,9H),0.01(d,J＝5.5Hz,6H).

Dissolving compound BB-1c (1.50g,0.46mmol) in tetrahydrofuran (30mL), adding TBAF (7.0mL,7.0mmol, 1.0M), reacting at room temperature until TLC detection reaction is complete, adding saturated aqueous ammonium chloride (100mL), extracting with ethyl acetate (100 mL. times.2), washing with saturated saline, drying over anhydrous magnesium sulfate, and separating with flash chromatography

BB-1(0.61g, yield 63%) was isolated.

And step 3: synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-oxa-7-azaspiro [3.5] nonane (1)

Compound 1d (50mg,0.11mmol) was dissolved in dichloromethane (3mL), trifluoroacetic acid (1mL) was added, stirring was carried out at room temperature until completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give a crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give Compound 1(4mg, 10% yield).

MS(ESI)m/z:372.2[M+H]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.57(d,J＝2.1Hz,1H),8.03(dd,J＝2.5,8.7Hz,1H),7.31-7.07(m,1H),7.02(d,J＝8.8Hz,1H),5.00(d,J＝3.7Hz,2H),4.36(s,4H),3.72(d,J＝1.8Hz,2H),3.11-2.91(m,4H),2.08-1.87(m,4H).

Example 2

Synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-methyl-7-azaspiro [3.5] nonan-2-ol (2)

Step 1: synthesis of 7- [ 6-chloropyridin-3-yl) sulfonyl ] -2-methyl-7-azaspiro [3.5] nonan-2-ol (2b)

Compound 1a (1.17g,4.72mmol) was dissolved in dichloromethane (10mL) and triethylamine (1.90g,18.9mmol) and compound 6-chloropyridine-3-sulfonyl chloride (1a) (1.00g,4.72mmol) were added and the reaction was monitored by LC-MS for completion at room temperature. Filtering, and separating with flash chromatography

Isolation gave 2b (0.80g, 51% yield).

MS(ESI)m/z:331.1[M+H]⁺

¹H NMR(400MHz,CDCl₃)δ8.76(dd,J＝0.6,2.4Hz,1H),8.00(dd,J＝2.6,8.3Hz,1H),7.65-7.48(m,1H),3.07-2.93(m,4H),1.92-1.85(m,4H),1.84-1.79(m,2H),1.72-1.64(m,2H),1.37(s,3H).

Step 2: synthesis of 7- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-methyl-7-azaspiro [3.5] nonan-2-ol (2c)

Compound 2b (322mg,0.98mmol) was dissolved in N-methylpyrrolidone (4mL) and triethylamine (150mg,1.46mmol) was added, followed by cooling to 0 ℃ with an ice-water bath. A solution of BB-1(200mg,0.98mmol) in tetrahydrofuran (8mL) and a solution of sodium tert-butoxide (141mg,1.46mmol) in dimethyl sulfoxide (2mL) were added. The reaction was monitored by LC-MS at room temperature for completion, water (10mL) was added, extracted with ethyl acetate (10mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered and the filtrate was evaporated to dryness to give crude 2c (200mg, yield 41%).

MS(ESI)m/z:500.2[M+H]⁺。

And step 3: synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-methyl-7-azaspiro [3.5] nonan-2-ol (2)

Dissolve compound 2c (200mg,0.40mmol) in dichloromethane (3mL), add trifluoroacetic acid (1mL), stir at room temperature until LC-MS monitors the reaction completion, concentrate the reaction to give the crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give Compound 2(31mg, 14% yield).

MS(ESI)m/z:400.2[M+H]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.58(d,J＝2.3Hz,1H),8.05(dd,J＝2.6,8.8Hz,1H),7.47-7.13(m,1H),7.05(d,J＝8.7Hz,1H),5.02(d,J＝3.5Hz,2H),3.83(d,J＝2.0Hz,2H),2.96(br d,J＝4.3Hz,4H),1.91-1.81(m,4H),1.80-1.73(m,2H),1.71-1.63(m,2H),1.30(s,3H)

Example 3

Synthesis of methyl 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-carboxylate (3)

Step 1: synthesis of 7- [ 6-chloropyridin-3-yl) sulfonyl ] -2-methyl-7-azaspiro [3.5] nonan-2-ol (3b)

Compound 3a (1.68g,5.66mmol) was dissolved in dichloromethane (10mL) and triethylamine (1.53g,15.1mmol), DMAP (50mg,0.38mmol) and the compound 6-chloropyridine-3-sulfonyl chloride (1a) (1.00g,4.72mmol) were added and reacted at room temperature until LC-MS monitored that the reaction was complete. Using a flash chromatography separation instrument

Isolation was carried out to give 3b (0.83g, yield 61%).

MS(ESI)m/z:359.1[M+H]⁺¹H NMR(400MHz,CDCl₃)δ8.75(d,J＝2.5Hz,1H),7.98(dd,J＝2.5,8.3Hz,1H),7.51(d,J＝8.3Hz,1H),3.67(s,3H),3.09-2.95(m,5H),2.03-1.93(m,4H),1.72(td,J＝5.6,13.7Hz,4H).

Wherein 3a can be prepared from commercial 3a-1 by the following operations:

dissolve 3a-1(2.00g,7.43mmol) in acetone (20mL) add potassium carbonate (3.08g,22.3mmol) and methyl iodide (2.31mL,37.1mmol) and react at room temperature until LC-MS monitors the reaction completion. Filtering, and separating with flash chromatography

3a-2(1.40g, 67% yield) was isolated as a colorless oil.

MS(ESI)m/z:284.2[M+H]⁺

¹H NMR(400MHz,CDCl₃)δ3.71-3.67(m,3H),3.38-3.33(m,2H),3.31-3.26(m,2H),3.09(quin,J＝8.8Hz,1H),2.06(d,J＝8.8Hz,4H),1.60-1.56(m,2H),1.55-1.50(m,2H),1.45(s,9H).

Compound 3a-2(1.00g,3.53mmol) was dissolved in dichloromethane (10mL), trifluoroacetic acid (3.4mL) was added, the reaction was stirred at room temperature until completion of TLC (ethyl acetate/petroleum ether ═ 1: 3) and the reaction solution was concentrated to give a crude product. The crude product was used directly in the next reaction.

MS(ESI)m/z:184.2[M+H]⁺

Step 2: synthesis of methyl 7- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-carboxylate (3c)

Compound 3b (350mg,0.98mmol) was dissolved in N-methylpyrrolidone (4mL) and triethylamine (150mg,1.46mmol) was added, followed by cooling to 0 ℃ with an ice-water bath. A solution of BB-1(200mg,0.98mmol) in tetrahydrofuran (8mL) and a solution of sodium tert-butoxide (141mg,1.46mmol) in dimethyl sulfoxide (2mL) were added. The reaction was allowed to proceed to LC-MS at room temperature to monitor completion of the reaction, water (10mL) was added, extraction was performed with ethyl acetate (10mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude product 3c (500mg, yield 97%). MS (ESI) M/z 428.0[ M-Boc]⁺。

And 3, step 3: synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-carboxylic acid (3)

Compound 3c (500mg,0.95mmol) was dissolved in dichloromethane (4.5mL), trifluoroacetic acid (1.5mL) was added, stirring was carried out at room temperature until completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give a crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give Compound 3(46mg, yield 9%).

MS(ESI)m/z:428.2[M+H]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.56(d,J＝2.3Hz,1H),8.03(dd,J＝2.5,8.8Hz,1H),7.39-7.14(m,1H),7.05(d,J＝8.8Hz,1H),5.01(d,J＝3.5Hz,2H),3.83(s,2H),3.63(s,3H),3.15-3.04(m,1H),3.01-2.96(m,2H),2.95-2.89(m,2H),2.03-1.90(m,4H),1.77-1.71(m,2H),1.68-1.62(m,2H)

Examples 4 and 5

2- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-azaspiro [3.3] heptane (4) and

synthesis of 2- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-azaspiro [3.3] heptane (5)

Step 1: synthesis of 2- [ 6-chloropyridin-3-yl) sulfonyl ] -2-azaspiro [3.3] heptane (4b)

Compound 4a (38mg,2.59mmol) was dissolved in dichloromethane (10mL) and triethylamine (716mg,7.07mmol) and the compound 6-chloropyridine-3-sulfonyl chloride (1a) (0.50g,2.36mmol) were added and the reaction was allowed to react at room temperature until LC-MS monitored for completion. Mixing reactant with silica gel directly, and separating with rapid chromatographic separator

Isolation was carried out to give 4b (0.20g, yield 31%) as a white solid. MS (ESI) M/z 273.1[ M + H ]]⁺

¹H NMR(400MHz,CDCl₃)δ8.83(d,J＝2.3Hz,1H),8.07(dd,J＝2.4,8.3Hz,1H),7.55(d,J＝8.3Hz,1H),3.79(s,4H),2.09-2.03(m,4H),1.85-1.76(m,2H).

Step 2: synthesis of 2- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-azaspiro [3.3] heptane (4c)

Compound 4b (53mg,0.20mmol) was dissolved in N-methylpyrrolidone (1mL) and triethylamine (30mg,0.29mmol) was added, followed by cooling to 0 ℃ with an ice-water bath. A solution of BB-1(40mg,0.20mmol) in tetrahydrofuran (2mL) and a solution of sodium tert-butoxide (28mg,0.29mmol) in dimethyl sulfoxide (0.5mL) were added. The reaction was allowed to proceed to LC-MS at room temperature to monitor completion of the reaction, water (10mL) was added, extraction was performed with ethyl acetate (10mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude 4c (50mg, yield 58%). MS (ESI) M/z 442.2[ M + H ]]⁺。

And 3, step 3: synthesis of 2- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-azaspiro [3.3] heptane (4) and 2- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -2-azaspiro [3.3] heptane (5)

Compound 4c (50mg,0.11mmol) was dissolved in dichloromethane (3mL), trifluoroacetic acid (1mL) was added, stirring was carried out at room temperature until completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give a crude product. The crude product was directly separated by preparative high performance liquid chromatography to give Compound 4(11mg, yield 28%) and Compound 5(3mg, yield 6%). MS (ESI) M/z 342.1[ M + H ]]⁺

Compound 4:

MS(ESI):m/z 342.1[M+H]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.63(d,J＝2.3Hz,1H),8.11(dd,J＝2.4,8.8Hz,1H),7.38-7.12(m,1H),7.08(d,J＝8.7Hz,1H),5.03(d,J＝3.3Hz,2H),3.79(d,J＝1.5Hz,2H),3.74(s,4H),2.10-1.97(m,4H),1.88-1.72(m,2H).

compound 5:

MS(ESI):m/z 342.1[M+H]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.64(d,J＝2.3Hz,1H),8.11(dd,J＝2.5,8.7Hz,1H),7.38-6.96(m,2H),5.19(d,J＝2.6Hz,2H),3.75(s,4H),3.69(d,J＝2.7Hz,2H),2.14-1.94(m,4H),1.87-1.69(m,2H).

example 6

Synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-formylethylamine (6)

Step 1: synthesis of 7- [ 6-chloropyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-formylethylamine (6b)

Compound 6a (2.20g,7.07mmol) was dissolved in dichloromethane (10mL) and triethylamine (2.62mL,18.9mmol), DMAP (60mg,0.47mmol) and compound 6-chloropyridine-3-sulfonyl chloride (1a) (1.00g,4.71mmol) were added and the reaction was monitored for completion by LC-MS at room temperature. Using a flash chromatography separation instrument

Isolation gave 6b (1.00g, yield 57%). MS (ESI) M/z 272.1[ M + H]⁺

¹H NMR(400MHz,CDCl₃)δ8.73(d,J＝2.5Hz,1H),7.97(dd,J＝2.5,8.3Hz,1H),7.50(d,J＝8.3Hz,1H),5.41(br s,1H),3.31-3.22(m,2H),3.06-2.93(m,4H),2.90-2.84(m,1H),2.05-1.86(m,4H),1.75-1.68(m,4H),1.10(t,J＝7.3Hz,3H).

Wherein 6a can be prepared using commercially available 3a-1 by the following procedure:

synthesis of 7-tert-butyl formate-7-azaspiro [3.5] nonane-2-formylethylamine (6a-1)

3a-1(2.50g,9.28mmol) was dissolved in DMF (20mL), cooled to 0 deg.C, HATU (4.24g,11.1mmol), HOBt (1.38g,10.2mmol) and DIEA (3.07mL,18.60mmol) were added, and diethylamine hydrochloride (1.14g,13.90mmol) was slowly added with stirring. The reaction was monitored by LC-MS for completion at room temperature. Quenching the reaction with water (20mL), extracting with ethyl acetate (20mLx3), combining the organic phases, washing with saturated brine, drying the organic phase with anhydrous sodium sulfate, filtering, mixing the filtrate with silica gel, and separating by flash chromatography

Isolation gave 6a-1(2.51g, 95% yield).

MS(ESI):m/z 297.2[M+H]⁺

¹H NMR(400MHz,DMSO-d₆)δ4.02(q,J＝7.1Hz,1H),3.28-3.12(m,4H),3.08-2.99(m,2H),2.94-2.84(m,1H),2.73-2.63(m,4H),1.85(d,J＝8.6Hz,4H),1.37(s,9H),0.98(t,J＝7.2Hz,3H).

Synthesis of 7-azaspiro [3.5] nonane-2-formylethylamine (6a)

Compound 6a-1(1.50g,5.06mmol) was dissolved in dichloromethane (15mL), trifluoroacetic acid (5mL) was added, the reaction was stirred at room temperature until completion of TLC (ethyl acetate/petroleum ether ═ 1: 2) and the reaction solution was concentrated to give a crude product (2.00 g). The crude product was used directly in the next reaction. MS (ESI) M/z 197.2[ M + H]⁺

And 2, step: synthesis of 7- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-formylethylamine (6c)

Compound 6b (360mg,0.97mmol) was dissolved in N-methylpyrrolidone (4mL) and triethylamine (0.20mL,1.45mmol) was added, followed by cooling to 0 ℃ with an ice water bath. A solution of BB-1(200mg,0.98mmol) in tetrahydrofuran (8mL) and a solution of sodium tert-butoxide (140mg,1.45mmol) in dimethyl sulfoxide (2mL) were added. The reaction was warmed to room temperature until completion of the reaction was monitored by LC-MS, water (10mL) was added, extraction was performed with ethyl acetate (10mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude 6c (507mg, yield 100%). MS (ESI) M/z 541.2[ M + H ]]⁺。

And step 3: synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-formylethylamine (6)

Compound 6c (523mg,0.97mmol) was dissolved in dichloromethane (3mL), trifluoroacetic acid (1mL) was added, stirring was carried out at room temperature until completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give a crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give Compound 6(18mg, 4% yield).

MS(ESI):m/z 441.1[M+H]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.55(d,J＝2.5Hz,1H),8.01(dd,J＝2.5,8.8Hz,1H),7.30-6.92(m,2H),4.98(d,J＝3.5Hz,3H),3.66(s,2H),3.15(q,J＝7.3Hz,2H),3.03-2.88(m,5H),1.89(d,J＝8.5Hz,4H),1.77-1.71(m,2H),1.68-1.62(m,2H),1.07(t,J＝7.2Hz,3H).

Example 7

8- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -1-oxo-2-methyl-2-8-diazaspiro [4.5] decane (7)

Step 1: synthesis of 8- [ 6-chloropyridin-3-yl) sulfonyl ] -7-azaspiro [3.5] nonane-2-formylethylamine (7b)

Dissolve 11a (2.00g,7.07mmol) in dichloromethane (10mL) add triethylamine (2.62mL,18.9mmol), DMAP (60mg,0.47mmol) and the compound 6-chloropyridine-3-sulfonyl chloride (1a) (1.00g,4.71mmol) and react at room temperature to LC-MS monitor reaction completion. Mixing reactant with silica gel directly, and separating with rapid chromatographic separator

Isolation gave 7b (1.20g, 56% yield). MS (ESI) M/z 344.1[ M + H ]]⁺

¹H NMR(400MHz,CDCl₃)δ8.77(d,J＝2.5Hz,1H),7.99(dd,J＝2.5,8.3Hz,1H),7.50(d,J＝8.3Hz,1H),3.53-3.48(m,2H),3.30(t,J＝6.9Hz,2H),3.50-3.03(m,2H),2.81(s,3H),2.01-1.92(m,2H),1.84(t,J＝6.9Hz,2H),1.65-1.55(m,2H).

Wherein 7a can be prepared using commercial 7a-1 by the following procedure:

synthesis of 8-carboxylic acid tert-butyl ester-1-oxo-2-8-diazaspiro [4.5] decane (7a-2)

Compound 7a-1(2.50g,9.83mmol) was dissolved in DMF (25mL), cooled to 0 ℃ with an ice water bath, 60% sodium hydride (0.79mg,19.70mmol) was added, and the reaction was continued at 0 ℃ for 10 minutes. Slowly adding methyl iodide (1.22mL,19.7mmol), maintaining the temperature at 0 ℃ for reaction until the reaction is completely monitored by LC-MS, adding water (200mL), extracting with ethyl acetate (150mL x3), combining organic phases, washing with saturated salt water, drying with anhydrous magnesium sulfate, filtering, evaporating filtrate to obtain a crude product, directly mixing the reactant with silica gel, and separating by flash chromatographyInstrument for measuring the position of a moving object

Isolation gave 7a-2(2.51g, 95% yield).

MS(ESI):m/z 169.2[M-Boc]⁺。

¹H NMR(400MHz,CDCl₃)δ4.00(br s,2H),3.32(t,J＝7.0Hz,2H),2.98(br t,J＝11.2Hz,2H),2.86(s,3H),1.97(br t,J＝6.5Hz,2H),1.89-1.89(m,1H),1.89-1.82(m,1H),1.46(s,9H),1.37(d,J＝13.1Hz,2H).

Synthesis of 1-oxo-2-8-diazaspiro [4.5] decane (7a)

Compound 7a-2(1.00g,3.73mmol) was dissolved in dichloromethane (10mL), trifluoroacetic acid (3mL) was added, stirring was carried out at room temperature until the completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give the crude product (2.00 g). The crude product was used directly in the next reaction. MS (ESI) M/z 169.1[ M + H]⁺

Step 2: synthesis of 8- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -1-oxo-2-methyl-2-8-diazaspiro [4.5] decane (7c)

Compound 7b (335mg,0.97mmol) was dissolved in N-methylpyrrolidone (4mL) and triethylamine (0.20mL,1.45mmol) was added, cooled to 0 ℃ with an ice-water bath. A solution of BB-1(200mg,0.98mmol) in tetrahydrofuran (8mL) and a solution of sodium tert-butoxide (140mg,1.45mmol) in dimethyl sulfoxide (2mL) were added. The reaction was warmed to room temperature until completion of the reaction was monitored by LC-MS, water (80mL) was added, extraction was performed with ethyl acetate (40 mL. times. 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude 7c (499 mg). MS (ESI) M/z 413.0[ M-Boc]⁺。

And step 3: synthesis of 8- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -1-oxo-2-methyl-2-8-diazaspiro [4.5] decane (7)

Compound 7c (499mg,0.97mmol) was dissolved in dichloromethane (4.5mL), trifluoroacetic acid (1.5mL) was added, stirring was carried out at room temperature until the reaction was complete as monitored by LC-MS, and the reaction solution was concentrated to give the crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give compound 7(68mg, yield 15%).

MS(ESI):m/z 413.1[M+H]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.61-8.50(m,2H),8.05(dd,J＝2.5,8.8Hz,1H),7.32-7.09(m,1H),7.03(d,J＝8.8Hz,1H),5.00(d,J＝3.5Hz,2H),3.76(s,2H),3.62-3.55(m,2H),3.36-3.33(m,1H),2.81-2.71(m,5H),1.91-1.82(m,4H),1.60-1.51(m,2H).

Example 8

Synthesis of 8- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -1-oxo-2-oxa-8-azaspiro [4.5] decane (8)

Step 1: synthesis of 8- [ 6-chloropyridin-3-yl) sulfonyl ] -1-oxo-2-oxa-8-azaspiro [4.5] decane (8b)

Compound 8a (723mg,3.77mmol) was dissolved in dichloromethane (10mL) and triethylamine (1.43g,14.1mmol) and the compound 6-chloropyridine-3-sulfonyl chloride (1a) (1.00g,4.71mmol) were added and the reaction was monitored by LC-MS at room temperature for completion. Mixing reactant with silica gel directly, and separating with rapid chromatographic separator

Isolation gave 8b (0.50g, 40% yield).

MS(ESI):m/z 331.1[M+H]⁺

¹H NMR(400MHz,CDCl₃)δ8.79(d,J＝2.1Hz,1H),8.22(dd,J＝2.4,8.4Hz,1H),7.83(d,J＝8.4Hz,1H),4.23(br t,J＝7.0Hz,2H),3.48(br d,J＝12.1Hz,2H),2.90-2.68(m,2H),2.07(br t,J＝6.9Hz,2H),1.71(br d,J＝4.5Hz,4H).

Step 2: synthesis of 8- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -1-oxo-2-oxa-8-azaspiro [4.5] decane (8c)

Compound 8b (65mg,0.20mmol) was dissolved in N-methylpyrrolidone (1mL) and triethylamine (30mg,0.29mmol) was added, followed by cooling to 0 ℃ with an ice-water bath. A solution of BB-1(40mg,0.20mmol) in tetrahydrofuran (2mL) and a solution of sodium tert-butoxide (28mg,0.29mmol) in dimethyl sulfoxide (0.5mL) were added. The reaction was warmed to room temperature until completion of the reaction was monitored by LC-MS, water (10mL) was added, extraction was performed with ethyl acetate (10mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude 8c (50mg, yield 58%). MS (ESI) M/z 400.1[ M-Boc]⁺。

And 3, step 3: synthesis of 8- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -1-oxo-2-oxa-8-azaspiro [4.5] decane (8)

Compound 8c (50mg,0.10mmol) was dissolved in dichloromethane (4.5mL), trifluoroacetic acid (1.5mL) was added, the reaction was stirred at room temperature for 3 to LC-MS to monitor completion of the reaction, and the reaction was concentrated to give the crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give Compound 8(10mg, yield 23%).

MS(ESI):m/z 400.0[M+H]⁺

¹H NMR(400MHz,CDCl₃)δ8.61(d,J＝2.4Hz,1H),8.08(dd,J＝2.5,8.7Hz,1H),7.43-7.15(m,1H),7.07(d,J＝8.8Hz,1H),5.03(d,J＝3.5Hz,2H),4.31(t,J＝7.0Hz,2H),3.84(d,J＝2.0Hz,2H),3.44-3.37(m,2H),3.06-3.01(m,2H),2.15(t,J＝7.0Hz,2H),2.03-1.87(m,2H),1.79-1.76(m,2H).

Example 9

6- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid methyl ester (9) and

synthesis of methyl 6- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylate (10)

Step 1: synthesis of methyl 6- [ 6-chloropyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylate (9b)

Compound 9a (1.00g,3.53mmol) was dissolved in dichloromethane (10mL) and triethylamine (1.90g,18.90mmol) and the compound 6-chloropyridine-3-sulfonyl chloride (1a) (1.00g,4.71mmol) were added and the reaction was monitored at room temperature by LC-MS for completion. Mixing reactant with silica gel directly, and separating with rapid chromatographic separator

Isolation gave 9b (0.80g, 49% yield). MS (ESI) M/z 331.1[ M + H ]]⁺

¹H NMR(400MHz,CDCl3)δ8.78(d,J＝2.0Hz,1H),8.01(dd,J＝2.5,8.4Hz,1H),7.53(d,J＝8.3Hz,1H),3.67(s,3H),3.35-2.94(m,4H),1.95-1.79(m,2H),1.67-1.61(m,1H),1.58-1.51(m,2H),1.13(t,J＝5.1Hz,1H),0.92(dd,J＝4.8,8.2Hz,1H).

Wherein 9a can be prepared from commercial 9a-1 by the following operations:

synthesis of 6-tert-butylcarbonate-6-azaspiro [2.5] octane-1-carboxylic acid methyl ester (9a-2)

Dissolve 9a-1(1.50g,5.88mmol) in acetone (15mL) add potassium carbonate (1.62g,11.80mmol) and methyl iodide (1.97mL,11.80mmol) and react at room temperature until LC-MS monitors the reaction completion. Filtering to remove insoluble substances, directly mixing the filtrate with silica gel, and separating with flash chromatography

Isolation gave 9a-2(1.40g, 89% yield). MS (ESI) M/z 270.1[ M + H ]]⁺

¹H NMR(400MHz,CDCl3)δ3.68(s,3H),3.56-3.38(m,3H),3.28(d,J＝7.5Hz,1H),1.70(dt,J＝4.3,7.2Hz,2H),1.58-1.54(m,1H),1.49-1.36(m,11H),1.18(t,J＝5.0Hz,1H),0.95(dd,J＝4.6,8.0Hz,1H).

Synthesis of 6-azaspiro [2.5] octane-1-carboxylic acid methyl ester trifluoroacetate (9a)

Compound 9a-2(1.00g,5.17mmol) was dissolved in dichloromethane (10mL), trifluoroacetic acid (3mL) was added, the reaction was monitored by TLC (ethyl acetate/petroleum ether ═ 1: 3) at room temperature and the reaction solution was concentrated to give the crude product. The crude product was used directly in the next reaction. MS (ESI) M/z 170.2[ M + H ]]⁺

Step 2: synthesis of methyl 6- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylate (9c)

Compound 9b (336mg,0.98mmol) was dissolved in N-methylpyrrolidone (4mL) and triethylamine (148mg,1.46mmol) was added, followed by cooling to 0 ℃ with an ice-water bath. A solution of BB-1(200mg,0.98mmol) in tetrahydrofuran (8mL) and a solution of sodium tert-butoxide (140mg,1.46mmol) in dimethyl sulfoxide (2mL) were added. The reaction was warmed to room temperature until completion of the reaction was monitored by LC-MS, water (80mL) was added, extraction was performed with ethyl acetate (40mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude product 9c (200mg, yield 46%). MS (ESI) M/z 414.0[ M-Boc]⁺。

And step 3: 6- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid methyl ester (9) and 6- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid methyl ester (10)

Compound 9c (200mg,0.39mmol) was dissolved in dichloromethane (4.5mL), trifluoroacetic acid (1.5mL) was added, stirring was carried out at room temperature until the reaction was monitored by LC-MS, and the reaction solution was concentrated to give the crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give compound 9(130mg, yield 58%) compound 10(5mg, yield 2%). MS (ESI) M/z 413.9[ M + H ]]⁺

Compound 9:

MS(ESI):m/z 413.9[M+H]⁺

¹H NMR(400MHz,CDCl3)δ8.60(d,J＝2.2Hz,1H),8.07(dd,J＝2.5,8.7Hz,1H),7.42-7.13(m,1H),7.06(d,J＝8.8Hz,1H),5.03(d,J＝3.5Hz,2H),3.84(d,J＝2.0Hz,2H),3.62(s,3H),3.16-3.05(m,3H),2.98(br dd,J＝3.7,7.3Hz,1H),2.03-1.73(m,2H),1.66-1.51(m,3H),1.05(s,2H).

compound 10:

MS(ESI):m/z 413.9[M+H]⁺

¹H NMR(400MHz,CDCl3)δ8.61(d,J＝2.3Hz,1H),8.07(dd,J＝2.5,8.7Hz,1H),7.31-6.98(m,2H),5.18(d,J＝2.4Hz,2H),3.72(d,J＝2.4Hz,2H),3.62(s,3H),3.17-3.07(m,3H),3.02-2.85(m,1H),1.92-1.81(m,2H),1.63-1.55(m,3H),1.08-0.93(m,2H).

example 10

6- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxamidemethanamine trifluoroacetate (11) and

synthesis of 6- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxamidine trifluoroacetate (12)

Step 1: synthesis of 6- [ 6-chloropyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid (11a)

Compound 9b (1.10g,3.19mmol) was dissolved in tetrahydrofuran (5mL) and water (1mL), lithium hydroxide hydrate (268mg,6.38mmol) was added and the reaction was allowed to react at room temperature until LC-MS monitored completion. Adjusting the pH value to 5-7 with 1N hydrochloric acid, extracting with ethyl acetate, combining organic phases, washing with saturated salt solution, drying with anhydrous sodium sulfate, filtering, and evaporating filtrate to obtain a crude product 11a (1.00 g). MS (ESI) M/z 331.1[ M + H ]]⁺。

Step 2: synthesis of 6- [ 6-chloropyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-methanamide (11b)

Compound 11a (1.00g,3.02mmol) was dissolved in DMF (10mL) and HOBt (449mg,3.33mmol), DIPEA (781mg,6.05mmol), HATU (1.38g,3.33mmol) and methylamine hydrochloride (370mg,4.54mmol) were added. The reaction was monitored at room temperature to completion by LC-MS. Adding water, extracting with ethyl acetate, mixing organic phases, washing with saturated salt solution, drying with anhydrous sodium sulfate, filtering, evaporating filtrate to obtain crude product, and separating by column chromatography to obtain compound 11b (0.60g, 58%). MS (ESI) M/z 344.1[ M + H ]]⁺。

And 3, step 3: synthesis of 6- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-formylmethylamine (11c)

Compound 11b (0.67g,1.95mmol) was dissolved in NMP (5mL) and triethylamine (296mg,2.92mmol) was added, cooled to 0 ℃ with an ice-water bath, followed by addition of a solution of BB-1(400mg,1.95mmol) in tetrahydrofuran (10mL) and a solution of sodium tert-butoxide (281mg,2.92mmol) in DMSO (2.5 mL). The reaction was warmed to room temperature until completion of the reaction was monitored by LC-MS, water (30mL) was added, ethyl acetate (30mL × 2) was extracted, the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude 11c (500 mg). MS (ESI) M/z 413.2[ M-Boc + H]⁺。

And step 3: synthesis of 6- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-ylmethylamine trifluoroacetate (11) and 6- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-ylmethylamine trifluoroacetate (12)

Compound 11c (300mg,0.58mmol) was dissolved in dichloromethane (3.0mL), trifluoroacetic acid (1.0mL) was added, stirring was carried out at room temperature until the reaction was monitored by LC-MS and the reaction was concentrated to give the crude product. The crude product was directly separated by preparative HPLC to give Compound 11(127mg, 53% yield) MS (ESI) M/z 413.2[ M + H ]]⁺

¹H NMR(400MHz,METHANOL-d₄)δ8.59(d,J＝2.5Hz,1H),8.06(dd,J＝2.5,8.8Hz,1H),7.47-7.12(m,1H),7.06(d,J＝8.8Hz,1H),5.02(d,J＝3.5Hz,2H),3.85(s,2H),3.19(ddd,J＝3.8,7.1,11.2Hz,1H),3.13-2.95(m,3H),2.69(s,3H),1.79(t,J＝5.5Hz,2H),1.66-1.41(m,3H),1.03(t,J＝4.9Hz,1H),0.77(dd,J＝4.6,8.1Hz,1H)。

And Compound 12(9mg, yield 3%). MS (ESI) M/z 413.2[ M + H]⁺

¹H NMR(400MHz,METHANOL-d₄)δ8.60(d,J＝2.1Hz,1H),8.53(br s,1H),8.07(dd,J＝2.4,8.7Hz,1H),7.24-6.93(m,1H),5.18(d,J＝2.3Hz,2H),3.68(d,J＝2.4Hz,2H),3.25-2.96(m,4H),2.68(s,3H),1.80(br t,J＝5.4Hz,2H),1.68-1.31(m,3H),1.04(t,J＝4.9Hz,1H),0.77(dd,J＝4.5,8.0Hz,1H)。

Example 11

6- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid trifluoroacetate (13) and

synthesis of 6- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid trifluoroacetate salt (14)

Step 1: synthesis of 6- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid (11a)

Compound 9c (0.90g,1.75mmol) was dissolved in tetrahydrofuran (5mL) and water (1mL) and lithium hydroxide hydrate (0.15g,3.51mmol) was added and reacted at room temperature until LC-MS monitored reaction completion. Adjusting the pH value to 5-7 with 1N hydrochloric acid, extracting with ethyl acetate, combining organic phases, washing with saturated salt solution, drying with anhydrous sodium sulfate, filtering, and evaporating the filtrate to dryness to obtain a crude product 13a (800mg, yield 91%). MS (ESI) M/z 444.1[ M-tBut [)]⁺。

Step 2: synthesis of 6- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid trifluoroacetate (13) and 6- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -6-azaspiro [2.5] octane-1-carboxylic acid trifluoroacetate (14)

Compound 13a (500mg,1.00mmol) was dissolved in dichloromethane (3.0mL), trifluoroacetic acid (1.0mL) was added, stirring was carried out at room temperature until completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give a crude product. The crude product was directly separated by preparative HPLC to give compound 13(127mg, 30% yield.) MS (ESI) M/z 400.1[ M + H ]]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.60(d,J＝2.0Hz,1H),8.07(dd,J＝2.5,8.8Hz,1H),7.39-7.15(m,1H),7.06(d,J＝8.8Hz,1H),5.03(d,J＝3.3Hz,2H),3.84(d,J＝1.8Hz,2H),3.22-3.04(m,3H),3.02-2.83(m,1H),1.98-1.85(m,2H),1.66-1.47(m,3H),1.11-0.84(m,2H).

And 14(30mg, yield 7%). MS (ESI) M/z 400.1[ M + H ]]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.61(d,J＝2.3Hz,1H),8.08(dd,J＝2.5,8.8Hz,1H),7.32-6.77(m,2H),5.18(d,J＝2.5Hz,2H),3.72(d,J＝2.5Hz,2H),3.20-2.93(m,4H),1.95-1.78(m,2H),1.75-1.45(m,3H),1.04(t,J＝4.9Hz,1H),0.93(dd,J＝4.6,7.9Hz,1H)

Example 12

7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,5-a ] pyrazine trifluoroacetate (15) and

synthesis of 7- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,5-a ] pyrazine trifluoroacetate (16)

Step 1: synthesis of 7- [ 6-chloropyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,5-a ] pyrazine (15b)

Compound 15a (527g,4.24mmol) was dissolved in dichloromethane (10mL) and triethylamine (0.86g,8.49mmol) and the compound 6-chloropyridine-3-sulfonyl chloride (1a) (0.90g,4.24mmol) were added and the reaction was allowed to react at room temperature until LC-MS monitored for reaction completion. Mixing reactant with silica gel directly, and separating with rapid chromatographic separator

Isolation was carried out to give 15b (0.80g, yield 62%) as a white solid. MS (ESI) M/z 300.1[ M + H]⁺

¹H NMR(400MHz,CDCl₃)δ8.86(d,J＝2.5Hz,1H),8.07(dd,J＝2.5,8.4Hz,1H),7.91(s,1H),7.55(d,J＝8.4Hz,1H),4.54(s,2H),4.32(t,J＝5.4Hz,2H),3.79-3.73(m,2H).

Step 2: synthesis of 7- [6- [ -2- (((tert-butoxycarbonyl) amino) methyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,5-a ] pyrazine (15c)

Compound BB-1(684mg,3.34mmol) was dissolved in tetrahydrofuran (10mL), cooled to 0 ℃ in an ice-water bath, and 60% sodium hydride (227mg,6.68mmol) was added thereto, followed by reaction at 0 ℃ for 10 minutes. 15b (1.00g,3.34mmol) was added. The reaction was warmed to room temperature until completion of the reaction was monitored by LC-MS, water (30mL) was added, extraction was performed with methylene chloride (30mL × 2), the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated to dryness to give crude 15c (500 mg). MS (ESI) M/z 469.2[ M + H ]]⁺。

And step 3: synthesis of 7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,5-a ] pyrazine trifluoroacetate (15) and 7- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,5-a ] pyrazine trifluoroacetate (16)

Compound 15c (500mg,1.71mmol) was dissolved in dichloromethane (3mL), trifluoroacetic acid (1mL) was added, stirring was carried out at room temperature until completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give a crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give compound 15(94mg, yield 23%).

MS(ESI):m/z 369.1[M+H]⁺。

¹H NMR(400MHz,METHANOL-d4)δ8.72(d,J＝2.1Hz,1H),8.16(dd,J＝2.6,8.8Hz,1H),7.96(s,1H),7.40-7.10(m,1H),7.05(d,J＝8.8Hz,1H),5.19-4.97(m,2H),4.54(s,2H),4.27(t,J＝5.6Hz,2H),3.89-3.73(m,4H).

And compound 16(42mg, yield 10%). MS (ESI) M/z 369.1[ M + H ]]⁺

¹H NMR(400MHz,METHANOL-d4)δ8.73(d,J＝2.3Hz,1H),8.17(dd,J＝2.5,8.8Hz,1H),7.96(s,1H),7.33-6.91(m,2H),5.18(d,J＝2.5Hz,2H),4.54(s,2H),4.27(t,J＝5.5Hz,2H),3.80(t,J＝5.5Hz,2H),3.71(d,J＝2.8Hz,2H).

Example 13

7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2] oxadiazolo [1,2-a ] pyrazine trifluoroacetate (17) and

the procedure for synthesizing 7- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,2-a ] pyrazine trifluoroacetate (18) was similar to example 11, and the corresponding procedure was carried out using 5,6,7, 8-tetrahydroimidazo [1,2-a ] pyrazine hydrochloride instead of the compound 15a to obtain compound 17(335mg, yield 53%).

MS(ESI):m/z 368.1[M+H]⁺。

¹H NMR(400MHz,METHANOL-d4)δ8.73(d,J＝2.2Hz,1H),8.18(dd,J＝2.6,8.8Hz,1H),7.61-7.49(m,2H),7.42-7.14(m,1H),7.09(d,J＝8.8Hz,1H),5.04(d,J＝3.2Hz,2H),4.69(s,2H),4.35(t,J＝5.4Hz,2H),3.84(d,J＝2.1Hz,2H),3.80-3.67(m,2H).

And Compound 18(22mg, yield 3%), MS (ESI): M/z 368.1[ M + H%]⁺。¹H NMR(400MHz,METHANOL-d4)δ8.75(d,J＝2.6Hz,1H),8.19(dd,J＝2.6,8.8Hz,1H),7.51(s,2H),7.32-6.95(m,2H),5.19(d,J＝2.3Hz,2H),4.68(s,2H),4.45-4.28(m,1H),4.34(t,J＝5.4Hz,1H),3.88-3.74(m,2H),3.72(d,J＝2.3Hz,2H)。

Example 14

2-methyl-7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [1,5-a ] pyrazine trifluoroacetate (19) and

2-methyl-7- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy]Pyridin-3-yl) sulfonyl]-5,6,7, 8-tetrahydro- [1,2,4]Triazolo [1,5-a ]]Synthesis of pyrazine trifluoroacetate (20) analogously to example 11 using 2-methyl-5, 6,7, 8-tetrahydro- [1,2,4]]Triazolo [1,5-a]Pyrazine substituted for 15a to obtain 19(72mg, yield 18%), MS (ESI): M/z 383.1[ M + H ]]⁺，¹H NMR(400MHz,METHANOL-d4)δ8.71(d,J＝2.0Hz,1H),8.16(dd,J＝2.6,8.8Hz,1H),7.45-7.12(m,1H),7.05(d,J＝8.8Hz,1H),5.02(dd,J＝0.7,3.7Hz,2H),4.48(s,2H),4.18(t,J＝5.6Hz,2H),3.83(s,2H),3.78(t,J＝5.6Hz,2H),2.31(s,3H)。

And compound 20(22mg, yield 54%). MS (ESI) M/z 383.1[ M + H [ ]]⁺，¹H NMR(400MHz,METHANOL-d4)δ8.72(d,J＝2.1Hz,1H),8.17(dd,J＝2.6,8.8Hz,1H),7.39-6.93(m,2H),5.18(d,J＝2.7Hz,2H),4.48(s,2H),4.18(t,J＝5.5Hz,2H),3.78(t,J＝5.6Hz,2H),3.71(d,J＝2.3Hz,2H),2.31(s,3H)。

Example 15

7- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -5,6,7, 8-tetrahydro- [1,2,4] triazolo [4,3-a ] pyrazine trifluoroacetate (21) and

7- [6- [ (Z) -2- (aminomethyl) -3-fluoroallyloxy group]Pyridin-3-yl) sulfonyl]-5,6,7, 8-tetrahydro- [1,2,4]Triazolo [4,3-a]Synthesis of pyrazine trifluoroacetate (22) analogously to example 11 using 5,6,7, 8-tetrahydro- [1,2,4]]Triazolo [4,3-a]Pyrazine substituted for 15a to obtain 21(35mg, yield 7%), MS (ESI): M/z 369.1[ M + H ]]⁺，¹H NMR(400MHz,METHANOL-d4)δ8.72(d,J＝2.1Hz,1H),8.48(s,1H),8.18(dd,J＝2.6,8.9Hz,1H),7.46-7.14(m,1H),7.09-7.00(m,1H),5.08-4.99(m,2H),4.57(s,2H),4.24(t,J＝5.6Hz,2H),3.83(d,J＝1.2Hz,2H),3.72(t,J＝5.6Hz,2H)。

And Compound 22(18mg, yield 4%), MS (ESI): M/z 369.1[ M + H%]⁺，¹H NMR(400MHz,METHANOL-d4)δ8.74(d,J＝2.4Hz,1H),8.48(s,1H),8.19(dd,J＝2.5,8.9Hz,1H),7.40-6.87(m,2H),5.18(d,J＝2.7Hz,2H),4.58(s,2H),4.24(t,J＝5.6Hz,2H),3.89-3.67(m,4H)。

Example 16

3-methyl-8- [6- [ (E) -2- (aminomethyl) -3-fluoroallyloxy ] pyridin-3-yl) sulfonyl ] -1,3, 8-triazaspiro [4.5] decane-2, 4-dione trifluoroacetate (23) and

3-methyl-8- [6- [ (Z) -2- (aminomethyl)) -3-fluoroallyloxy]Pyridin-3-yl) sulfonyl]-1,3, 8-triazaspiro [4.5]Synthesis of decane-2, 4-dione trifluoroacetate (24) by a procedure similar to example 14, substituting 23a for 15a gave compound 23(388mg, yield 68%), MS (ESI): M/z 428.0[ M + H428.0 [)]⁺，¹H NMR(400MHz,METHANOL-d4)δ8.60(d,J＝2.4Hz,1H),8.07(dd,J＝2.6,8.8Hz,1H),7.39-7.13(m,1H),7.06(d,J＝8.7Hz,1H),5.02(d,J＝3.7Hz,2H),3.82(d,J＝2.1Hz,2H),3.60-3.44(m,2H),3.31(td,J＝1.6,3.3Hz,3H),3.10-3.07(m,2H),2.05-2.00(m,2H),1.81-1.76(m,2H)。

And 24(11mg, yield 2%), MS (ESI) M/z 428.0[ M + H]⁺，¹H NMR(400MHz,METHANOL-d4)δ8.61(d,J＝2.1Hz,1H),8.07(dd,J＝2.6,8.8Hz,1H),7.18-6.87(m,2H),5.17(d,J＝2.4Hz,2H),3.62-3.50(m,4H),3.10-2.99(m,2H),2.90(s,3H),2.06-2.02(m,2H),1.80-1.75(m,2H)。

Wherein 23a can be prepared using commercially available 23a-1 by the following procedure:

synthesis of 3-methyl-8-tert-butoxycarbonyl-1, 3, 8-triazaspiro [4.5] decane-2, 4-dione (23a-2)

Compound 23a-1(5.90g,99.80mmol) was dissolved in DMSO (40mL) and cooled to 0 deg.C, and methyl iodide (1.16g,8.17mmol) and potassium carbonate (3.08g,22.30mmol) were added. The reaction was warmed to room temperature until completion was monitored by LC-MS. Diluted with 200mL of water and extracted with ethyl acetate (100mLx 2). The organic phases were combined, washed with saturated brine and dried over anhydrous sodium sulfate. Separating by column chromatography to obtain compound 23a-2(1.74g, yield 83%) MS (ESI) with M/z 184.4[ M + H ]]⁺。¹H NMR(400MHz,CDCl₃)δ6.95(s,1H),4.12-3.87(m,2H),3.20(ddd,J＝2.9,10.5,13.5Hz,2H),3.02(s,3H),2.08-1.93(m,2H),1.63(s,2H),1.47(s,9H)。

Synthesis of 3-methyl-1, 3, 8-triazaspiro [4.5] decane-2, 4-dione (23a)

Compound 23a-2(3.00g,13.90mmol) was dissolved in dichloromethane (30mL), trifluoroacetic acid (10mL) was added, stirring was carried out at room temperature until completion of the reaction was monitored by LC-MS, and the reaction solution was concentrated to give the crude product. The crude product was directly isolated by preparative high performance liquid chromatography to give compound 23a (1.13g), which was used directly in the next reaction.

Biological evaluation

The present disclosure is further described and explained below in conjunction with test examples, which are not meant to limit the scope of the present disclosure.

Test example 1: evaluation of inhibitory Activity of rhVAP-1 enzyme

(1) Instrument consumable and reagent

Multifunctional enzyme-linked immunosorbent assay (MD, FlexStation3), black impermeable bottom 96-1L plate (Corning), rhVAP-1(PeproTech)

(2) Self-preparation of compound concentration gradient solution

An appropriate amount of test compound was dissolved in DMSO to 10mM and stored. Before the experiment, a proper amount of 10mM stock solution of the compound to be tested is diluted to 1mM solution by DMSO, then the stock solution is diluted by 3-fold gradient by DMSO, 10 concentration gradients are obtained, and then the stock solution is diluted by 100-fold by PBS to prepare 10X series concentration compound solutions.

(3) Preparation of enzyme solution

An appropriate amount of protein diluent was added to the rhVAP-1 powder to give 1mg/mL of mother liquor for storage. The enzyme solution was diluted with PBS before the experiment to 4 Xconcentration.

(4) Preparation of 2x concentration substrate mixture

Weighing a proper amount of benzylamine, adding PBS to dissolve the benzylamine to obtain a 200mM benzylamine solution, adding 2mM Amplex Red mother liquor and 500U/mL HRP mother liquor, and diluting the mixture with the PBS to obtain substrate mixed liquor with the concentration of 2 x.

(5) Test method

First, 10. mu.L of compound solutions of different concentrations, 25. mu.L of 4 XrhVAP-1 enzyme solution and 15. mu.L of PBS were added to a 96-well plate, mixed by shaking, and incubated at 37 ℃ for 30 min. Then adding 50 mu L of 2x substrate mixed liquor into each hole, immediately using a microplate reader for detection, exciting light at 565nm, emitting light at 590nm, detecting the fluorescence intensity of each hole for 5 min/time for 25min, and calculating the inhibition rate according to the following formula:

V(RFU/min)＝(F_t(RFU)-F₀(RFU))/(time (min))

Inhibition ratio (%) - < 100% to V_cmpd(RFU/min)/V_max(RFU/min)x 100％

V: rate of change of fluorescence F_tFluorescence reading F at time t₀: an initial fluorescence reading; time: a duration t; v_cmpdThe rate of change of fluorescence V of the test compound_maxMax pore fluorescence change rate.

(6) Fitting dose-effect curve

The log value of the concentration is taken as an X axis, the percent inhibition rate is taken as a Y axis, and a dose-effect curve is fitted by adopting the log (inhibitor) vs. response-Variable slope of the GraphPad Prism 5 analysis software, so as to obtain the IC of each compound on the enzyme activity₅₀The value is obtained.

In vitro inhibition of VAP-1 enzyme Activity by the examples of the present disclosure the IC determined by the above assay₅₀The values are shown in Table 1.

TABLE 1

Remarking:

(PXS-4728A)；

(BI-38-Z)。

test example 2: selectivity of MAO-A/B enzyme

(1) Instrument consumable and reagent

Microplate reader (Perkin Elmer, EnVision), 384 well plates (Perkin Elmer), centrifuge (Eppendorf), MAO-GloTM (Promega), MAO-A (active motif) and MAO-B (active motif).

(2) Self-preparation of compound concentration gradient solution

The test compound was dissolved in DMSO to 10mM and stored, and then diluted with DMSO in 4-fold gradient for a total of 6 concentration gradients.

(3) Preparation of enzyme solution

The MAO-A/B stock was diluted with MAO-A/B assay buffer to A2 Xconcentration of enzyme solution.

(4) Preparation of 2x concentration substrate mixture

The MAO-A/B substrate mixture mother liquor was diluted to 2 Xconcentration with MAO-A/B assay buffer.

(5) Test method

200nL of compound solutions or solvents with different concentrations, 10. mu.L of 2xMAO-A/B enzyme solution, were added to A384-well plate, centrifuged at 1000rpm for 60s, shaken, mixed, and incubated at room temperature for 15 min. The reaction was then initiated by adding 10. mu.L of 2 Xsubstrate mix per well. The 384 well plates were centrifuged at 1000rpm for 60s, shaken well and incubated at room temperature for 60 min. The reaction was stopped by adding 20. mu.L of a termination detection solution, centrifuging at 1000rpm for 60 seconds, and shaking and mixing. Standing for 30min, and reading with a microplate reader.

The inhibition rate was calculated according to the following formula:

suppression ratio (%) (Signal _ Max-Signal _ sample)/(Signal _ Max-Signal _ min) x 100

(6) Fitting dose-effect curve

Examples of the disclosure in vitro inhibition of MAO-A and MAO-B enzyme Activity by the above assays, IC₅₀The values are shown in Table 2.

TABLE 2

As can be seen from the above tables 1 and 2, the compounds of the present disclosure have good inhibitory activity against rhVAP-1 enzyme, and show excellent selective inhibitory effect against rhVAP-1 enzyme compared to monoamine oxidase (MAO). From the above tables 1 and 2, it is clear that the compounds of the present disclosure do not cause side effects due to inhibition of rhAOC1 enzyme or MAO enzyme while treating and/or preventing SSAO/VAP-1 enzyme-related diseases.

Test example 3: in vivo pharmacokinetic study in mice

1. Abstract (abstract)

The drug concentration in plasma of mice administered with compound 16 by gavage was measured by LC/MS/MS method using mice as test animals. The pharmacokinetic behavior of the compounds of the disclosure was studied in mice and evaluated for their pharmacokinetic profile.

2. Test protocol

2.1 test drugs

Compound 16

2.2 test animals

C57 mice 9, female, were divided into 3 groups on average, purchased from shanghai jestie laboratory animals ltd, animal production license number: SCXK (Shanghai) 2013-0006

2.3 pharmaceutical formulation

A defined amount of the compound was weighed and dissolved in 1% hydroxyethylcellulose (w/v) and 0.25% Tween 80 to give a colorless clear solution at 1 mg/ml.

2.4 administration

C57 mice were fasted overnight and then gavaged at 10mg/kg for each dose and at 0.1ml/10g for each volume.

3. Operation of

The mouse is administrated by gastric lavage, blood is collected for 0.1ml before and after administration at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0 and 24.0 hours, the blood is placed in a heparinized test tube, after centrifugation is carried out for 10 minutes at 3500 rpm, blood plasma is separated, and the blood plasma is stored at the temperature of minus 20 ℃.

Determination of the content of the test compound in the plasma of mice administered with different concentrations of the drug by injection: 25 mu L of mouse plasma at each moment after administration is taken, 50 mu L (100ng/mL) of camptothecin (China institute for biological products) as an internal standard solution and 200 mu L of acetonitrile are added, vortex mixing is carried out for 5 minutes, centrifugation is carried out for 10 minutes (4000 rpm), and 4 mu L of supernatant fluid of a plasma sample is taken for LC/MS/MS analysis.

4. Pharmacokinetic parameter results

TABLE 3

In addition, in a caco-2 permeability experiment, ER (equal to 24) of the compound indicates that the brain permeability is very low, and the compound has very good safety.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof

R³and R⁴Independently selected from hydrogen, deuterium, C_1-6An alkyl group optionally substituted with one or more R^A1Each independently substituted, R^A1Selected from halogen, deuterium, hydroxy, nitro, cyano or amino;

R⁵and R⁶Independently selected from hydrogen, deuterium, C_1-6An alkyl group optionally substituted with one or more R^A2Each independently substituted, R^A2Selected from halogen, deuterium, hydroxy, nitro, cyano or amino;

R⁷and R⁸Independently selected from hydrogen, deuterium, C_1-6An alkyl group optionally substituted with one or more R^A3Each independently substituted, R^A3Selected from halogen, deuterium, hydroxy, nitro, cyano or amino;

ring A is selected from a 4-to 6-membered heterocyclic ring, said ring A optionally substituted with one or more R^A4Substituted; r^A4Each independently selected from halogen, deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -SR^1a、-S(O)R^1a、-S(O)₂R^1a、-NR^1a(R^1b)、-(CH₂)_oCOR^1a、-(CH₂)_oNHCOR^1a、-(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCOR^1a；

R^1aOr R^1bEach independently selected from hydrogen, deuterium, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl being optionally substituted by one or more groups selected from halogen, deuterium, hydroxy, oxo, nitro, cyano or amino;

ring B is selected from a 3-to 6-membered carbocyclic ring, a 3-to 6-membered heterocyclic ring, or a 5-to 6-membered heteroaromatic ring, said ring B optionally substituted with one or more R^A5Substituted; r^A5Each independently selected from halogen, deuterium, oxo (═ O), hydroxy, nitro, cyano, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl, heteroaryl, -SR^2a、-S(O)R^2a、-S(O)₂R^2a、-NR^2a(R^2b)、-(CH₂)_pCOR^2a、-(CH₂)_pNHCOR^2a、-(CH₂)_pCONR^2a(R^2b)、-(CH₂)_pOCONR^2a(R^2b)、-N(CH₂)_pCONR^2a(R^2b)、-N(CH₂)_pCOR^2a；

R^2aOr R^2bEach independently selected from hydrogen, deuterium, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, said alkyl, alkoxy, cycloalkyl or heterocycloalkyl being optionally substituted by one or more groups selected from halogen, deuterium, hydroxy, oxo, nitro, cyano or amino;

and ring a is connected to ring B in a fused or spiro ring form;

X¹、X²、X³and X⁴Each is independent of othersIs selected from-CH-or-N-and is not simultaneously-N-;

o and p are each selected from integers between 0 and 3.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹Selected from hydrogen, R²Selected from fluorine; or R¹Selected from fluorine, R²Selected from hydrogen, further preferably R¹Selected from F, R²Selected from hydrogen.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R⁵、R⁶、R⁷And R⁸Selected from hydrogen.

4. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from

Further, ring A is optionally substituted with 1 to 3R^A4Substituted, R^A4As defined in claim 1.

5. A compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein

Is selected from

Wherein n and m are each independently selected from integers between 0 and 4, the sum of n and m is not more than 4, and Y is selected from-NH-, O, -CH₂-or-S-, further

By 1 to 3R^A4Or R^A5Substituted, R^A4、R^A5As defined in claim 1.

6. The method of any one of claims 1 to 5A compound or a pharmaceutically acceptable salt thereof, wherein

Is selected from

Preference is given to

Further, the method can be used for preparing a novel material

By 1 to 3R^A4Or R^A5Substituted, R^A4、R^A5As defined in claim 1; or

Is selected from

Preference is given to

Further, the method can be used for preparing a novel material

Is 1 to 2 of R^A4Or R^A5Substituted, R^A4、R^A5As in claimAs defined in claim 1.

7. The compound of any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein R^A4Each independently selected from halogen, deuterium, oxo (═ O), hydroxy, cyano or amino, or R^A4Each independently selected from C_1-6Alkyl or C_1-6Alkoxy, or R^A4Each independently selected from-NR^1a(R^1b)、-(CH₂)_oCOR^1a、-(CH₂)_oNHCOR^1a、-(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCONR^1a(R^1b)、-N(CH₂)_oCOR^1a，o、R^1aOr R^1bAs defined in claim 1.

8. The compound of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, wherein R^A5Each independently selected from halogen, deuterium, oxo (═ O), hydroxy, cyano or amino, or R^A5Each independently selected from oxo (═ O), hydroxy, halogen, C_1-6Alkyl or C_1-6Alkoxy, or R^A5Each independently selected from C_3-6Cycloalkyl, 3-to 6-membered heterocycloalkyl, aryl or heteroaryl, or R^A5Each independently selected from oxo (═ O), hydroxy, C_1-6Alkyl, -NR^2a(R^2b)、-(CH₂)_pCOR^2a、-(CH₂)_pNHCOR^2a、-(CH₂)_pCONR^2a(R^2b)、-(CH₂)_pCONR^2a(R^2b)、-N(CH₂)_pCONR^2a(R^2b)、-N(CH₂)_pCOR^2a，p、R^2aOr R^2bAs defined in claim 1, or R^A5Each independently selected from- (CH)₂)_pCOR^2aOr- (CH)₂)_pCONR^2a(R^2b)，p、R^1aOr R^1bAs defined in claim 1, or R^A5Each independently selected from fluorine and oxygenGeneration, C_1-3Alkyl, -COOH, -COC_1-3Alkoxy, -CONHC_1-3An alkoxy group.

9. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from a 5-membered heteroaromatic ring, preferably

Further, ring B is optionally substituted with 1 to 3R^A5Substituted, R^A5As defined in claim 1.

10. A compound according to any one of claims 1 to 4 or 9, or a pharmaceutically acceptable salt thereof, wherein

Is selected from

Which is further substituted by 1 to 3R^A4Or R^A5Substituted, R^A4、R^A5As defined in claim 1.

11. The compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein X²Is selected from-N-, X¹、X³、X⁴Is selected from-CH-; or X³Is selected from-N-, X¹、X²、X⁴Is selected from-CH-.

12. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I is

13. The compound of any one of claims 1-12, or a pharmaceutically acceptable salt thereof, wherein R³Selected from hydrogen or C_1-6Alkyl radicalPreferably hydrogen, methyl or ethyl; or R³Is selected from C_1-6Alkyl, preferably methyl, ethyl or propyl, optionally substituted by one or more R^A1Substituted, R^A1Each independently selected from halogen, deuterium, hydroxy, nitro, cyano or amino.

14. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound of formula I is selected from

Wherein

Including the E or Z configuration; further, the compounds of the formula I are preferably

15. A pharmaceutical composition comprising a therapeutically effective amount of at least one compound of any one of claims 1-14, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

16. Use of a compound according to any one of claims 1 to 14 or a pharmaceutical composition according to claim 15 for the manufacture of a medicament for the prevention and/or treatment of a condition associated with SSAO or SSAO/VAP-1.