CN109803967B - Dihydropyrimidine compound and preparation method and application thereof - Google Patents

Abstract

The invention provides a dihydropyrimidine compound shown as formula (I) or a pharmaceutically acceptable salt, a tautomer, an enantiomer or a diastereoisomer thereof, wherein R is₁、R₂、R₃R and A are as defined in the description and claims. The invention also provides a preparation method of the compound shown in the formula (I) and application of the compound in medicines for treating or preventing diseases infected by hepatitis B virus.

Description

Dihydropyrimidine compounds and preparation method and use thereof

technical field

The present invention relates to a dihydropyrimidine compound and its use as a medicine. Such compounds have therapeutic and preventive effects on hepatitis B, especially as hepatitis B virus (HBV) inhibitors, treating HBV infection by targeting the HBV capsid. The present invention also relates to processes for the preparation of such compounds.

Background technique

Viral hepatitis B is a disease caused by hepatitis B virus (HBV), mainly inflammatory lesions of the liver, and can cause multiple organ damage. Hepatitis B virus, referred to as hepatitis B virus, is a DNA virus belonging to the family hepadnavividae. It can cause acute or persistent/progressive chronic disease. Hepatitis B is widespread in countries around the world, affecting more than 400 million people, especially in the Asia-Pacific region. A small number of these patients can be transformed into cirrhosis or liver cancer. The anti-HBV nucleoside (acid) drugs currently on the market include lamivudine, telbivudine, entecavir, tenofovir disoproxil, and clavudine. The disadvantages of this type of drug are: the course of treatment is not fixed, virus resistance is prone to occur, and it is easy to relapse after drug withdrawal. These shortcomings prevent patients from being cured.

Deres et al. reported heteroaromatic substituted dihydropyrimidine (HAP) compounds represented by Bay41-4109 and Bay3905493, which can inhibit HBV replication by preventing the formation of normal nucleocapsid. Bay41-4109 showed better drug metabolism parameters in clinical studies (Deres K. et al., Science, 299 (2003), 893-896). The study of its mechanism of action found that the heteroaromatic ring-substituted dihydropyrimidines interacted with the 113-143 amino acid residues of the core protein to change the angle between the dimers that form the nucleocapsid, resulting in the formation of incompatibility. Stable swelling of the nucleocapsid, accelerated degradation of the core protein (Biochem. Pharmacol. 66 (2003), 2273-2279).

There is still a need for new compounds that are effective against viruses, especially as drugs for the treatment and/or prevention of hepatitis B.

SUMMARY OF THE INVENTION

The present invention provides a compound represented by formula (I), or a pharmaceutically acceptable salt or tautomer or enantiomer or diastereomer thereof:

in:

R ₁ is phenyl, wherein said phenyl is optionally further substituted by one or more substituents selected from halogen, C _1-6 alkyl;

R ₂ is selected from hydrogen or C _1-4 alkyl;

A is a bond, -O-, -S- or -N(R ₅ )-;

R ₅ is hydrogen or C _1-4 alkyl;

R ₃ is selected from heteroaryl, preferably thiazolyl, oxazolyl, imidazolyl, thienyl, phenyl or pyridyl, which may be further selected from one or more of halogen, methylene (=CH ₂ ), oxo (=O), alkyl, alkoxy, cyano, hydroxyl, nitro, alkylamino, mercapto, amino, aryl, haloalkyl, alkylsulfonyl, arylalkyl, heteroaryl radical, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, cycloalkyl, cycloalkylalkyl, trifluoromethyl, trifluoromethoxy, haloalkyl substituted aryl, halogen substituted aryl substituted with a substituent of a trifluoromethanesulfonyl group or a trifluoromethanesulfonyl group;

R is the group shown below:

R ₆ is each independently -(CR ₇ R _7a ) _m -OH, -(CR ₇ R _7a ) _n -C(=O)OR ₈ , -(CR ₇ R _7a ) _n -C(=O)-NH -S(O) ₂ -R ₉ , -(CR ₇ R _7a ) _n -C(O)-NR ₁₀ R _10a , -(CR ₇ R _7a ) _n -C(=O)O-(CR ₇ R _7a ) _n -OC(=O)OR ₈ , -S(=O) _p OR ₈ , -(CR ₇ R _7a ) _n -S(=O) _p N(R ₈ ) ₂ , -(CR ₇ R _7a ) _n -C(=O)O-(CR ₇ R _7a ) _n -OC(=O)-R ₈ , -(CR ₇ R _7a ) _n -C(=O)O-(CR ₇ R _7a ) _n - C(=O)OR ₈ , -(CR ₇ R _7a ) _n -N(R ₈ ) ₂ , or -(CR ₇ R _7a ) _n -C(=O)N(R ₈ ) ₂ ;

R ₇ and R _7a are each independently hydrogen, halogen, alkyl or haloalkyl, wherein said alkyl, haloalkyl is optionally further selected from one or more of halogen, methylene (=CH ₂ ), oxygen substituted (=O), alkyl, alkoxy, cyano, hydroxyl, nitro, alkylamino, mercapto, amino, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, Heterocyclylalkyl, cycloalkyl, cycloalkylalkyl, trifluoromethyl, trifluoromethoxy, haloalkyl substituted aryl, halo substituted aryl or substituted with a trifluoromethanesulfonyl substituent , or R ₇ and R _7a and the carbon atoms to which they are attached together form a cycloalkyl or heterocyclyl;

R ₈ is each independently hydrogen, hydroxy, alkyl, alkoxy, alkyl-S(=O) _p- , aryl, heteroaryl, cycloalkyl, heterocyclyl, arylalkyl, heterocycle radical-S(=O) _p- , heteroaryl-S(=O) _p- , cycloalkyl-S(=O) _p- or aryl-S(=O) _p- , wherein the alkane radical, alkoxy, alkyl-S(=O) _p- , aryl, heteroaryl, cycloalkyl, heterocyclyl, arylalkyl, heterocyclyl-S(=O) _p- , heterocyclyl Aryl-S(=O) _p- , cycloalkyl-S(=O) _p- or aryl-S(=O) _p- is optionally further selected by one or more of halogen, methylene (= CH ₂ ), oxo (=O), alkyl, alkoxy, cyano, hydroxy, nitro, alkylamino, mercapto, amino, aryl, arylalkyl, heteroaryl, heteroarylalkyl , heterocyclyl, heterocyclylalkyl, cycloalkyl, cycloalkylalkyl, trifluoromethyl, trifluoromethoxy, haloalkyl substituted aryl, halogen substituted aryl, or trifluoromethanesulfonyl substituted by the substituent;

R ₉ is alkyl, alkoxy, aryl, heteroaryl, cycloalkyl, heterocyclyl or arylalkyl;

R ₁₀ and R _10a are each independently hydrogen, haloalkyl, cycloalkyl, alkyl or hydroxyalkyl, or R ₁₀ and R _10a together with the nitrogen atom to which they are attached form a heterocyclyl;

m is independently 1, 2, 3, 4;

n is each independently 0, 1, 2, 3, 4;

p is each independently 1 or 2.

In one embodiment of the present invention, R is preferably the substructural formula shown below:

In one embodiment of the present invention, R is preferably selected from:

In _one embodiment of the present invention, R1 is preferably selected from phenyl, optionally further substituted with one or more halogens.

In another embodiment of the present invention, R ₂ is preferably methyl or ethyl.

In another embodiment of the present invention, A is preferably -O-.

In another embodiment of the present invention, R ₃ is preferably thiazolyl, imidazolyl or pyridyl, wherein said thiazolyl, imidazolyl or pyridyl is optionally further selected by one or more selected from halogen, alkyl, Substituents of alkoxy, haloalkyl, alkylsulfonyl, and cycloalkyl are substituted.

In another embodiment of the present invention, R ₁ is preferably phenyl substituted with 2 halogens; R ₂ is preferably methyl or ethyl; A is preferably -O-; R ₃ is preferably selected from thiazolyl, imidazolyl or pyridine wherein said thiazolyl, imidazolyl or pyridyl is optionally further selected from one or more of halogen, methylene (=CH ₂ ), oxo (=O), alkyl, alkoxy, cyano group, hydroxyl, nitro, alkylamino, mercapto, amino, aryl, haloalkyl, alkylsulfonyl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, substituted by cycloalkyl, cycloalkylalkyl, trifluoromethyl, trifluoromethoxy, haloalkyl substituted aryl, halogen substituted aryl or trifluoromethanesulfonyl substituent; R is preferably one of the following shown groups:

R ₆ is each independently -(CR ₇ R _7a ) _n -C(=O)OR ₈ , -(CR ₇ R _7a ) _n -C(=O)O-(CR ₇ R _7a ) _n -OC(= O)-R ₈ or -(CR ₇ R _7a ) _n -C(=O)O-(CR ₇ R _7a ) _n -C(=O)OR ₈ ;

R ₇ and R _7a are each independently hydrogen, halogen, alkyl or haloalkyl, wherein said alkyl, haloalkyl is optionally further selected from one or more of halogen, methylene (=CH ₂ ), oxygen substituted (=O), alkyl, alkoxy, cyano, hydroxyl, nitro, alkylamino, mercapto, amino, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, Heterocyclylalkyl, cycloalkyl, cycloalkylalkyl, trifluoromethyl, trifluoromethoxy, haloalkyl substituted aryl, halo substituted aryl or substituted with a trifluoromethanesulfonyl substituent , or R ₇ and R _7a and the carbon atoms to which they are attached together form a cycloalkyl or heterocyclyl;

R ₈ is each independently hydrogen, hydroxy, alkyl, alkoxy, alkyl-S(=O) _p- , aryl, heteroaryl, cycloalkyl, heterocyclyl, arylalkyl, heterocycle radical-S(=O) _p- , heteroaryl-S(=O) _p- , cycloalkyl-S(=O) _p- or aryl-S(=O) _p- , wherein the alkane radical, alkoxy, alkyl-S(=O) _p- , aryl, heteroaryl, cycloalkyl, heterocyclyl, arylalkyl, heterocyclyl-S(=O) _p- , heterocyclyl Aryl-S(=O) _p- , cycloalkyl-S(=O) _p- or aryl-S(=O) _p- is optionally further selected by one or more of halogen, methylene (= CH ₂ ), oxo (=O), alkyl, alkoxy, cyano, hydroxy, nitro, alkylamino, mercapto, amino, aryl, arylalkyl, heteroaryl, heteroarylalkyl , heterocyclyl, heterocyclylalkyl, cycloalkyl, cycloalkylalkyl, trifluoromethyl, trifluoromethoxy, haloalkyl substituted aryl, halogen substituted aryl, or trifluoromethanesulfonyl substituted by the substituent;

n is each independently 0, 1, 2, 3, 4, preferably 0;

p is each independently 1 or 2.

More preferably, the compound formula (I) of the present invention is selected from:

(1) 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane- 5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(2) (S)-5-((6-(2-Chloro-4-fluorophenyl)-5-(ethoxycarbonyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine -4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid;

(3) 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-(morpholine-4-carbonyl)-5-azaspiro[2.4]heptan-5-yl ) methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(4) (S)-5-(((R)-6-(2-Chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-1,4 -dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid;

(5) 4-(2-Chloro-4-fluorophenyl)-2-(3,5-fluoropyridin-2-yl)-6-(((S)-6-((methylsulfonyl)amino) formyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylate;

(6) 4-(2-Chloro-4-fluorophenyl)-2-(3,5-fluoropyridin-2-yl)-6-((S)-5-azaspiro[2.4]heptane- 6-Carboxylic acid)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester;

(7) 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane- 5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate ethyl ester;

(8) 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-((cyclopropylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane -5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate ethyl ester;

(9) 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-(isopropylcarbamoyl)-5-azaspiro[2.4]heptan-5-yl ) methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(10) (S)-5-(((R)-6-(2-Chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(4-methylthiazol-2-yl) -3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid;

(11) (S)-5-(((R)-6-(2-Chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3,6 -dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid;

(12) (R)-Methyl-4-(2-chloro-3-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-aza spiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(13) (1R,3S,5R)-2-(((R)-6-(2-chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl) -3,6-dihydropyrimidin-4-yl)methyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid;

(14) (1R,3S,5R)-2-(((R)-6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(4-methylthiazole- 2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid;

(15) (R)-Methyl-4-(2-chloro-4-fluorophenyl)-6-(((S)-6-(isopropylcarbamoyl)-5-azaspiro[2.4 ]heptan-5-yl)methyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(16) (R)-methyl-4-(2-chloro-4-fluorophenyl)-6-(((1R,3S,5R)-3-((methylsulfonyl)carbamoyl)- 2-azabicyclo[3.1.0]hexane-2-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(17) (S)-5-(((R)-6-(2-Chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(1-methyl-1H-imidazole- 2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid;

(18) (R)-Methyl 4-(2-chloro-3-fluorophenyl)-6-(((S)-6-((cyclopropylsulfonyl)carbamoyl)-5-aza spiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(19) (1S,2S,5R)-3-(((R)-6-(2-chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl) -3,6-dihydropyrimidin-4-yl)methyl)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid;

(20) (R)-Methyl-4-(2-chloro-4-fluorophenyl)-6-(((S)-6-(hydroxymethyl)-5-azaspiro[2.4]heptane -5-yl)methyl)-2-(thipyridin-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(21) (R)-Methyl-4-(2-bromo-4-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-aza spiro[2.4]heptan-5-yl)methyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(22) (S)-5-(((R)-6-(2-Bromo-4-fluorophenyl)-5-(methoxycarbonyl)-2-(4-methylthiazol-2-yl )-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid;

(23) (1S,2S,5R)-3-(((R)-6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl) )-3,6-dihydropyrimidin-4-yl)methyl)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid;

(24) (R)-Methyl-4-(2-chloro-4-fluorophenyl)-2-(1-methyl-1H-imidazol-2-yl)-6-(((S)-6 -((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylate;

(25) (R)-Methyl-4-(2-bromo-4-fluorophenyl)-2-(4-methylthiazol-2-yl)-6-(((S)-6-(( phenylsulfonyl)carbamoyl)-5-azaspiro[2.4]hept-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(26) (R)-Methyl-4-(2-chloro-4-fluorophenyl)-6-(((S)-6-(methylcarbamoyl)-5-azaspiro[2.4] heptan-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(27) (R)-Methyl 4-(2-chloro-3-fluorophenyl)-6-(((S)-6-(dimethylaminomethyl)-5-azaspiro[2.4] heptan-5-yl)methyl)-2-(thipyridin-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester;

(28) (S)-5-(((R)-6-(2-bromo-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3, 6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid;

(29) (S)-((isopropoxycarbonyl)oxy)methyl 5-(((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-(thiazole- 2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylate methyl ester;

(30) (S)-(pivaloyloxy)methyl 5-(((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-(thiazol-2-yl) -3,6-Dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylate methyl ester.

The second aspect of the present invention provides a pharmaceutical composition comprising an effective dose of the compound of formula (I) or a pharmaceutically acceptable salt or tautomer or enantiomer or diastereomer thereof isomer.

Yet another aspect of the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt or tautomer or enantiomer or diastereomer thereof, or a pharmaceutical composition thereof, in the manufacture of a therapeutic or Use of medicines for the prevention of hepatitis B virus-infected diseases.

Yet another aspect of the present invention provides processes for the preparation of compounds of formula (I). The method comprises: reacting the compound represented by the formula d with the R-H compound or its salt in the presence of a base to obtain the compound of the formula (I):

wherein R ₁ , R ₂ , R ₃ , R and A are as defined above; wherein the base is preferably an organic base, more preferably triethylamine.

wherein the compound of formula d is prepared by the following method, including:

Step (1): the compound represented by formula a is reacted with the compound represented by formula b to obtain the compound represented by formula c:

Step (2): the compound shown in formula c is reacted with a bromination reagent to obtain the compound shown in formula d:

Wherein, R ₁ , R ₂ , R ₃ , R and A are as defined above; the bromination reagent in step (2) is preferably N-bromosuccinimide.

While certain embodiments of the present invention are now described in detail, the present invention is intended to cover all alternatives, modifications and equivalents, which are included within the scope of the present invention as defined by the claims. One skilled in the art should recognize that many methods and materials similar or equivalent to those described herein could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described herein. In the event that one or more of the incorporated literature, patents, and similar materials differs from or contradicts this application (including, but not limited to, defined terms, term applications, described techniques, etc.), the present invention prevail.

Unless otherwise defined, the terms used in the present invention have meanings generally accepted in the art. Further, some terms used in the present invention are defined as follows:

The term "comprising" is an open-ended expression, that is, it includes the contents specified in the present invention, but does not exclude other aspects.

The term "patient" as used herein may include humans (including adults and children) or other animals. In some embodiments, "patient" refers to a human.

"Alkyl" refers to a straight or branched chain saturated aliphatic hydrocarbon group. The alkyl group in this application is preferably a C _1-6 alkyl group, which means a saturated straight or branched chain alkyl group comprising 1 to 6 carbon atoms; the particularly preferred alkyl group in this application is a C _1-4 alkyl group , that is, a saturated straight or branched chain alkyl group of 1 to 4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, 1-butyl, 2-butyl and tert-butyl and the like.

"Alkoxy" refers to a group (alkyl-O-). wherein the alkyl group is as defined above. Preferred alkoxy groups are C _1-6 alkoxy groups, and particularly preferred alkoxy groups are C _1-4 alkoxy groups. The term _C1-6alkoxy includes methoxy, ethoxy, n-propoxy, isopropoxy, and the like.

The term "cycloalkyl" denotes a saturated carbocyclic ring containing 3 to 12 carbon atoms, especially 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. In particular, "cycloalkyl" is cyclopropyl, cyclopentyl, cyclohexyl.

"Haloalkyl" or "haloalkoxy" means an alkyl or alkoxy group substituted with one or more halogen atoms, the same or different. Preferred alkyl or alkoxy groups are as defined above. Examples include, but are not limited to: trifluoromethyl, trifluoroethyl, trifluoromethoxy.

"Cycloalkylalkyl" means that an alkyl group is substituted with one or more cycloalkyl groups. Examples include, but are not limited to: cyclopropylmethyl, cyclopropylethyl, cyclobutylmethyl, cyclopentylmethyl, and the like.

"Aryl" means monocyclic, bicyclic and tricyclic carbocyclic ring systems containing 6-14 ring atoms, wherein at least one ring system is aromatic, wherein each ring system contains 3-7 atoms A loop with one or more points of attachment to the rest of the molecule. Examples include, but are not limited to: phenyl, naphthyl, anthracene, and the like. Preferably, the aryl group is a carbocyclic ring system of 6-10 or 6-7 ring atoms.

"Heteroaryl" means monocyclic, bicyclic and tricyclic ring systems containing 5-14 ring atoms, wherein at least one ring system is aromatic and at least one ring system contains one or more selected from nitrogen, oxygen, A heteroatom of sulfur, where each ring system contains a ring of 5-7 atoms with one or more points of attachment to the rest of the molecule. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic". Examples include, but are not limited to: furyl, imidazolyl, 2-pyridyl, 3-pyridyl, thiazolyl, purinyl, quinolinyl. Preferably, the heteroaryl group is a ring system of 5-10 ring atoms.

"Heteroarylalkyl" means that a heteroaryl group is attached to the rest of the molecule through an alkyl group. Examples include, but are not limited to: pyridine-2-ethyl, thiazole-2-methyl, pyrimidine-2-propyl, and the like.

The term "amino" refers to a primary ( _-NH2 ), secondary (-NH-) or tertiary amino group

The term "carboxy" refers to the group -COOH.

The term "halogen" refers to fluorine, chlorine, bromine and iodine. In particular, halogen means fluorine, chlorine or bromine.

The term "cyano" refers to the group -CN.

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

The term "sulfonyl" refers to the group -S(O) _2- .

The term "carbonyl" refers to the group -C(=O)-.

The term "C _1-6 alkoxycarbonyl" refers to the group C _1-6 alkoxy-C(O)-, wherein said "C _1-6 alkoxy" is as defined above.

The term "aminocarbonyl" refers to the group amino-C(O)-, wherein said "amino" is as defined above.

The term "C _1-6 alkylsulfonyl" refers to the group C _1-6 alkyl-S(O) ₂ -, wherein said "C _1-6 alkyl" is as defined above.

The term "aminosulfonyl" refers to the group amino-S(O) _2- , wherein the "amino" is as defined above.

The term "heterocyclyl" refers to a non-aromatic saturated or partially unsaturated monocyclic, bicyclic or tricyclic ring system comprising 3 to 12 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. Wherein, the heterocyclyl group may be optionally substituted with one or more substituents described herein. Unless otherwise specified, heterocyclyl can be carbon or nitrogen, and _-CH2- groups can be optionally replaced by -C(=O)-. Ring sulfur atoms can optionally be oxidized to S-oxides. Preferably, the heterocyclic group is a non-aromatic saturated or partially unsaturated monocyclic, bicyclic or tricyclic ring system of 3-10, 3-6 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms.

The term "hydroxyalkyl" refers to a hydroxyalkyl group attached to the remainder of the molecule through a carbon atom, wherein the alkyl group is as defined above. The hydroxyalkyl group preferably includes a straight or branched chain alkyl group having 1 to 10 carbon atoms substituted with one or more hydroxy groups. More preferred hydroxyalkyl groups are lower hydroxyalkyl groups having 1-6 carbon atoms or 1-4 carbon atoms and one or more hydroxy groups, such as hydroxymethyl ( _-CH2OH ), hydroxyethyl (-CH ₂ CH ₂ OH) and hydroxypropyl (-CH ₂ CH ₂ CH ₂ OH) and the like.

The term "tautomer" refers to structural isomers of organic compounds that are readily interconverted by a chemical reaction known as tautomerization. This reaction usually results in the transfer of the form of a hydrogen atom or a proton with concomitant conversion of a single bond and an adjacent double bond, such as in compounds of general formula (I)

及其互变异构体

and its tautomers

The term "chiral" refers to a molecule that has the property of being non-superimposable with its mirror image; whereas "achiral" refers to a molecule that can overlap with its mirror image.

The term "enantiomers" refers to two non-superimposable but mirror-image isomers of a compound.

The term "diastereomer" refers to a stereoisomer having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers usually have different physical properties such as melting point, boiling point, spectral properties and reactivity. Diastereomeric mixtures can be separated by high resolution analytical procedures such as electrophoresis and chromatography, eg HPLC.

As described in the present invention, the compounds of the present invention may be optionally substituted with one or more substituents, such as compounds of the general formula above, specific compounds of the present invention, or specific examples, subclasses, as in the Examples, and a class of compounds encompassed by the present invention. It is to be understood that the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted". In general, the term "substituted" means that one or more hydrogen atoms in a given structure have been replaced with a specified substituent. Unless otherwise indicated, an optional substituent group may be substituted at various substitutable positions of the group. When a position in a given formula is not known to be substituted by one or more substituents selected from a particular group, the substituents may be substituted at each position identically or differently. The substituents can be, but are not limited to: fluorine, chlorine, bromine, iodine, oxo (=O), methylene (=CH ₂ ), alkyl, alkoxy, cyano, hydroxyl, nitro , alkylamino, mercapto, amino, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl, heterocyclylalkyl, cycloalkyl, cycloalkylalkyl, trifluoromethyl , trifluoromethoxy, haloalkyl substituted aryl, halogen substituted aryl or trifluoromethanesulfonyl and the like.

Furthermore, the term "pharmaceutically acceptable salts" refers to certain salts of the above-mentioned compounds which retain their original biological activity and which are suitable for medicinal use. The pharmaceutically acceptable salts of the compounds represented by formula (I) can be salts formed with suitable acids, suitable acids include inorganic and organic acids, such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid , ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, malic acid, maleic acid, mandelic acid, methanesulfonic acid, nitric acid, phosphoric acid, succinic acid , sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. Particularly preferred are hydrochloric acid, phosphoric acid or sulfuric acid.

Detailed ways

The following reactions are generally run under positive nitrogen pressure. Appropriate rubber stoppers are placed on the reaction vials, and the substrate can be injected through a syringe. Glassware is dried. The chromatographic column is a silica gel column. NMR data were determined on a BrukerAdvance 400 NMR instrument using _CDCl3 , d6- _DMSO or _CD3OD as solvent (reported in ppm) and with TMS (0 ppm) or chloroform (7.25 ppm) as reference standards. When multiplets are present, the following abbreviations are used: s (singlet, singlet), s, s (singlet, singlet, singlet, singlet), d (doublet, doublet), t (triplet, triplet), br (broadened, broad peak), dd (doublet of doublets, quartet), ddd (doublet of doublet of doublets, double double doublet), ddt (doublet of doublet of triplets, double double triplet), dddd (doublet of doublet of doublet of doublets, doublet of doublets), td (triplet of doublets, triplet of doublets), brs (broadened singlet, broad singlet). Coupling constant, expressed in Hertz (Hz).

Low resolution mass spectrometry (MS) data were determined by an Agilent 1100 Series LC-MS spectrometer. ESI sources are used in LC-MS spectrometers.

Compound purity was assessed by Agilent 1100 series high performance liquid chromatography (HPLC) with UV detection at 210nm and 254nm, Zorbax SB-C18 column, size 2.1X30mm, 4μm, 10min, flow rate 0.6ml/min, 5-95% (0.1% formic acid in acetonitrile) in (0.1% formic acid in water), column temperature maintained at 40°C.

The following abbreviations are used throughout this disclosure:

DCM or _{CH2Cl2 dichloromethane}

EtOAc or EA Ethyl acetate

THF tetrahydrofuran

CH ₃ OH or MeOH methanol

d ₆ -DMSO deuterated dimethyl sulfoxide

CDCl ₃ deuterated chloroform

CCl ₄ carbon tetrachloride

Boc tert-butoxycarbonyl

PE petroleum ether

K ₂ CO ₃ Potassium Carbonate

NaHCO ₃ Sodium Bicarbonate

Na ₂ SO ₄ Sodium Sulfate

KOAc Potassium Acetate

DIPEA N,N-Diisopropylethyl

amine

NBS N-bromosuccinimide

c concentration

g grams

v/v or v:v volume ratio

mole mole

mmol mmol

mL milliliter

L liter

h hours

t _1/2 half-life

AUC area under the curve

Vss Steady-state apparent distribution volume

product

CL or clearance clearance rate

F, absolute bioavailability

Dose

_Tmax peak time

_Cmax maximum concentration

hr*ng/mL plasma concentration*time

The starting materials and reagents of the present invention are all commercially available, and their suppliers are Aldrich Chemical Company, Alfa Chemical Company, Sinopharm Group, Lin'an Qingshan Chemical Reagent Factory, Jiangsu Huada Chemical Group and Hangzhou Chemical Reagent Co., Ltd. Commercially available starting materials and reagents were used without further purification unless otherwise indicated. In the examples described herein, unless otherwise indicated, all temperatures are in degrees Celsius (°C).

Example 1: 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane Preparation of methyl)-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

Step 1) Synthesis of compound 4-(2-chloro-4-fluorophenyl)-6-methyl-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester

The compound 2-(2-chloro-4-fluorobenzyl)-3-oxobutyric acid methyl ester (5.3 g, 20.7 mmol), 2-thiazolecarboxamidine hydrochloride (2.6 g, 15.9 mmol) was dissolved in three Fluoroethanol (70 ml), potassium acetate (3.1 g, 31.8 mmol) was added, nitrogen was replaced 3 times, and the reaction was heated under reflux at 80° C. overnight. After the reaction was completed, cooled, filtered with suction, concentrated the reaction solution, and the residue was separated and purified by column chromatography (n-heptane/ethyl ester (v/v)=9/1) to obtain 4-(2-chloro-4-) as a yellow solid Fluorophenyl)-6-methyl-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (4.4 g, 76%).

MS(ESI, pos.ion) m/z: 367[M+H] ⁺ ;

Step 2) Compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester synthesis

Under anhydrous operation, methyl 4-(2-chloro-4-fluorophenyl)-6-methyl-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate (500 mg , 1.37 mmol) was dissolved in CCl4 (15 ml), NBS (244 mg, 1.37 mmol) was added, and the reaction was carried out at room temperature for 1 h. After the reaction was completed, the reaction solution was concentrated, and the residue was separated and purified by column chromatography (n-heptane/ethyl ester (v/v)=4/1) to obtain a yellow solid (350 mg, 57%).

MS(ESI, pos.ion) m/z: 444,446[M+H] ⁺ ;

Step 3) Synthesis of compound (S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester

Compound (S)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (1.49 g, 6.18 mmol) was dissolved in THF (40 ml), CDI (2.2 g) was added , 13.6 mmol), the reaction was stirred at room temperature for 1 h, methylsulfonamide (2.35 g, 24.7 mmol), DBU (2.35 g, 15.5 mmol) were added, and the reaction was heated at 70° C. for 5 h. After the reaction was completed, cooled and concentrated the reaction solution, the residue was dissolved in DCM (50ml), washed with 1N HCl (50ml×2) and saturated brine (50ml×2) successively, the organic phase was concentrated, and the residue was subjected to column chromatography Isolation and purification (DCM/EA(v/v)=4/1) gave (S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane-5- as a white solid tert-Butyl formate (1.57 g, 80%).

MS(ESI, pos.ion) m/z: 319[M+H] ⁺ ;

Step 4) Synthesis of compound (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride

Under ice bath conditions, HCl gas (3.6 g, 98.7 mmol) was passed into THF (50 ml), and compound (S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4 ] Heptane-5-carboxylate tert-butyl ester (1.57 g, 4.94 mmol) was dissolved in the above solution, and the reaction was stirred at room temperature overnight. After the reaction was completed, the reaction solution was concentrated to obtain a white solid (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (0.95 g, 88%).

MS (ESI, pos.ion) m/z: 219 [M+H-HCl] ⁺ ;

Step 5) Compound 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane Synthesis of -5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (240 mg, 0.539mmol) was dissolved in CHCl3 (5ml), compound (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (176mg, 0.809mmol) was added , triethylamine (436mg, 4.31mmol), 50 ℃ heating reaction 5h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=19/1) to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)- 6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl) -1,4-Dihydropyrimidine-5-carboxylate methyl ester (70 mg, 22%).

MS(ESI, pos.ion) m/z: 583[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 8.47(br,1H), 7.90(s,1H), 7.61(s,s,1H), 7.34(m,1H), 7.21(m,1H), 7.16( m,1H),7.04(m,1H),6.16(s,s,1H),4.34(m,2H),3.69(s,3H),3.14(s,s,3H),2.22(m,2H) ,1.98(m,1H),1.34(m,2H),0.69(m,4H).

Example 2: (S)-5-((6-(2-Chloro-4-fluorophenyl)-5-(ethoxycarbonyl)-2-(thiazol-2-yl)-1,4-dihydro Preparation of pyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid

Step 1) Synthesis of compound 4-(2-chloro-4-fluorophenyl)-6-methyl-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester

2-(2-Chloro-4-fluorobenzyl)-3-oxobutyric acid ethyl ester (4.2 g, 15.8 mmol), 2-thiazolecarboxamidine hydrochloride (2.0 g, 12.1 mmol) were dissolved in trifluoro Ethanol (40 ml), potassium acetate (2.4 g, 24.3 mmol) was added, nitrogen was replaced three times, and the reaction was heated under reflux at 80° C. overnight. After the reaction was completed, cooled, filtered with suction, concentrated the reaction solution, and the residue was separated and purified by column chromatography (n-heptane/ethyl ester (v/v)=9/1) to obtain 4-(2-chloro-4-) as a yellow solid Fluorophenyl)-6-methyl-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate ethyl ester (2.6 g, 43%).

MS(ESI,pos.ion)m/z:381[M+H] ⁺

Step 2) Compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester synthesis

Dissolve 4-(2-chloro-4-fluorophenyl)-6-methyl-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (600 mg, 1.6 mmol) In CCl4 (5 ml), NBS (282 mg, 1.58 mmol) was added, and the reaction was carried out at room temperature for 1 h under nitrogen protection. After the reaction, the reaction solution was concentrated, and the residue was separated and purified by column chromatography (n-heptane/ethyl ester (v/v)=1/1) to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)- 6-(Bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (460 mg, 63%).

MS(ESI, pos.ion) m/z: 458, 460[M+H] ⁺

Step 3) Synthesis of compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride

Compound (S)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (5.0 g, 20.7 mmol) was dissolved in 50 ml of THF under ice-water bath conditions Then, HCl gas (8.5 g, 232.9 mmol) was bubbled into THF (50 ml) and the reaction was stirred at room temperature overnight. After completion of the reaction, the reaction solution was concentrated to obtain (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (3.5 g, 95%) as a white solid.

MS(ESI,pos.ion)m/z:142[M+H-HCl] ⁺

Step 4) Compound (S)-5-((6-(2-chloro-4-fluorophenyl)-5-(ethoxycarbonyl)-2-(thiazol-2-yl)-1,4-dihydro Synthesis of pyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid

Compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (100 mg, 0.218mmol) was dissolved in 5ml of CHCl3, added compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (87mg, 0.489mmol), triethylamine (476mg, 1.75mmol) , 40 ℃ heating reaction 5h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH(v/v)=9/1) to obtain a yellow solid (S)-5-((6-(2-chloro) -4-Fluorophenyl)-5-(ethoxycarbonyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4] Heptane-6-carboxylic acid (70 mg, 62%).

MS(ESI,pos.ion)m/z:520[M+H] ⁺

¹ H NMR (400MHz, MeOD): δppm 8.12(m, 2H), 7.55(m, 1H), 7.25(m, 1H), 7.12(m, 1H), 6.21(s, s, 1H), 4.05( m, 2H), 3.84(m, 1H), 3.52(m, 1H), 2.90(m, 1H), 2.48(m, 1H), 2.19(m, 1H), 2.00(m, 1H), 1.28(m ,2H),1.14(m,3H),0.72(m,4H).

Example 3: 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-(morpholine-4-carbonyl)-5-azaspiro[2.4]heptane-5- Preparation of methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (200 mg, 0.45mmol) was dissolved in CHCl3 (5ml), compound (S)-morpholine (5-azaspiro[2.4]heptan-6-yl)ethanone hydrochloride (189mg, 0.90mmol), triethylamine was added (365 mg, 3.6 mmol), heated at 45 °C for 1 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography using EA as a developing solvent to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)-6-(((S)-6 -(Morpholine-4-carbonyl)-5-azaspiro[2.4]heptane-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester (70 mg, 22%).

MS(ESI, pos.ion) m/z: 575[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 10.24(br,1H), 7.95(m,1H), 7.43(m,2H), 7.15(m,1H), 6.95(m,1H), 6.23(s, s,1H),4.19(m,1H),3.86-3.50(m,12H),2.98-2.70(m,1H),2.22(m,1H),1.93(m,1H),1.52(m,2H) ,0.69(m,4H).

Example 4: (S)-5-(((R)-6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-1, Preparation of 4-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (70 mg, 0.157mmol) was dissolved in CHCl3 (5ml), compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (42mg, 0.236mmol), triethylamine (127mg, 1.26mmol) were added ), heated at 50°C for 8h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH(v/v)=9/1) to obtain a yellow solid (6S)-5-(((R)-6- (2-Chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidin-4-yl)methyl)-5-aza Spiro[2.4]heptane-6-carboxylic acid (25 mg, 32%)

MS(ESI, pos.ion) m/z: 506[M+H] ⁺ ;

¹ H NMR (400MHz, MeOD): δppm 8.19(m, 2H), 7.47(m, 1H), 7.28(m, 1H), 7.14(m, 1H), 6.20(s, 1H), 3.68(s, 3H) ), 3.58(m, 2H), 2.90(m, 1H), 2.54(m, 1H), 2.22(m, 1H), 2.00(m, 1H), 1.40(m, 2H), 0.78(m, 4H) .

Example 5: 4-(2-Chloro-4-fluorophenyl)-2-(3,5-fluoropyridin-2-yl)-6-(((S)-6-((methylsulfonyl) Preparation of carbamoyl)-5-azaspiro[2.4]heptane-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylate methyl ester

The compound 6-(bromomethyl)-4-(2-chloro-4-fluorophenyl)-2-(3,5-difluoropyridin-2-yl)-1,4-dihydropyrimidine-5- Methyl carboxylate (150mg, 0.32mmol) was dissolved in CHCl3 (5ml), compound (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride was added Salt (70 mg, 0.32 mmol), triethylamine (259 mg, 2.56 mmol), heated at 40 °C for 4 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=19/1) to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)- 2-(3,5-Fluoropyridin-2-yl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane-5- (yl)methyl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (65 mg, 33%).

MS(ESI, pos.ion) m/z: 613[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.17(m,1H), 8.34(m,1H), 7.96(m,1H), 7.56(m,1H), 7.36(m,1H), 7.16(m, 1H),6.22(s,s,1H),4.12(m,1H),3.70(m,5H),3.24(s,s,3H),2.83(m,1H),2.42(m,1H),2.05 (m,1H),1.29(m,2H),0.69(m,4H).

Example 6: 4-(2-Chloro-4-fluorophenyl)-2-(3,5-fluoropyridin-2-yl)-6-((S)-5-azaspiro[2.4]heptane -6-Carboxylic acid)-1,4-dihydropyrimidine-5-carboxylate methyl ester

The compound 6-(bromomethyl)-4-(2-chloro-4-fluorophenyl)-2-(3,5-difluoropyridin-2-yl)-1,4-dihydropyrimidine-5- Methyl carboxylate (100mg, 0.21mmol) was dissolved in CHCl3 (5ml), compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (49mg, 0.27mmol) was added, tris Ethylamine (170 mg, 1.68 mmol) was stirred at room temperature for 5 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=9/1) to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)- 2-(3,5-Fluoropyridin-2-yl)-6-((S)-5-azaspiro[2.4]heptane-6-carboxylic acid)-1,4-dihydropyrimidine-5-carboxylate methyl ester (90 mg, 80%).

MS(ESI, pos.ion) m/z: 536[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 11.65(br,1H), 8.32(m,1H), 7.37(m,1H), 7.29(m,1H), 7.16(m,2H), 6.99(m, 1H), 6.16(s, s, 1H), 4.12(m, 1H), 3.80(m, 1H), 3.63(s, 3H), 2.42(m, 2H), 2.02(m, 1H), 1.32(m ,2H),0.69(m,4H).

Example 7: 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane Preparation of -5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate ethyl ester

Compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (100 mg , 0.22mmol) was dissolved in CHCl3 (5ml), compound (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (62mg, 0.28mmol) was added ), triethylamine (176 mg, 1.75 mmol), and the reaction was stirred at room temperature for 6 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=9/1) to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)- 6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl) -1,4-Dihydropyrimidine-5-carboxylate ethyl ester (60 mg, 46%).

MS(ESI, pos.ion) m/z: 597[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.17(s,1H), 8.51(s,s,1H), 7.63(s,s,1H), 7.22(m,1H), 7.16(m,1H), 7.01(m, 1H), 6.05(s, s, 1H), 4.14(m, 2H), 3.67(m, 1H), 3.48(m, 1H), 3.16(s, s, 3H), 2.72(m, 1H), 2.43(m, 1H), 2.12(m, 1H), 1.34(m, 2H), 1.15(m, 3H), 0.69(m, 4H).

Example 8: 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-((cyclopropylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptyl Preparation of ethyl alkyl-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

Compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (100 mg , 0.22mmol) was dissolved in CHCl3 (5ml), compound (S)-N-(cyclopropylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (69mg, 0.28 mmol), triethylamine (176 mg, 1.75 mmol), and the reaction was stirred at room temperature for 6 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=19/1) to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)- 6-(((S)-6-((cyclopropylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl )-1,4-dihydropyrimidine-5-carboxylic acid ethyl ester (70 mg, 51%).

MS(ESI, pos.ion) m/z: 623[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.50(br,1H), 8.57(s,s,1H), 7.60(s,s,1H), 7.27(m,1H), 7.14(m,1H), 7.03(m, 1H), 6.07(s, s, 1H), 4.14(m, 2H), 3.80-3.12(m, 2H), 2.80(m, 1H), 2.43(m, 1H), 2.20(m, 1H), 1.32(m, 2H), 1.21(m, 3H), 0.95(m, 4H), 0.71(m, 4H).

Example 9: 4-(2-Chloro-4-fluorophenyl)-6-(((S)-6-(isopropylcarbamoyl)-5-azaspiro[2.4]heptane-5- Preparation of methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (100 mg, 0.23mmol) was dissolved in CHCl3 (5ml), compound (S)-N-isopropyl-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (197mg, 0.90mmol), triethyl Amine (365 mg, 3.6 mmol) was heated at 45 °C for 1 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography using EA as a developing solvent to obtain a yellow solid 4-(2-chloro-4-fluorophenyl)-6-(((S)-6 -(Isopropylcarbamoyl)-5-azaspiro[2.4]heptane-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester (30 mg, 24%).

MS(ESI, pos.ion) m/z: 547[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.24(br,1H), 7.95(m,1H), 7.43(m,2H), 7.15(m,1H), 6.95(m,1H), 6.23(s, s,1H),4.39(m,1H),4.13-3.95(m,2H),3.75(s,3H),2.98-2.70(m,1H),2.22(m,1H),1.93(m,1H) ,1.22(m,6H),1.12(m,2H),0.74(m,4H).

Example 10: (S)-5-(((R)-6-(2-Chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(4-methylthiazol-2-yl )-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid preparation

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl The ester (80mg, 0.18mmol) was dissolved in CHCl3 (5ml), compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (64mg, 0.36mmol), triethylamine ( 146mg, 1.44mmol), heated at 45°C for 5h. After the reaction was completed, the reaction solution was cooled, concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=9/1) to obtain a yellow solid (20 mg, 22%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =520;

¹ H NMR (400MHz, CDCl ₃ ): δppm 11.33(br,1H), 7.64(s,1H), 7.33(m,1H), 7.31(m,1H), 7.03(m,1H), 6.17(s, 1H), 5.16(m, 1H), 4.58(m, 1H), 3.79(s, 3H), 3.52(m, 1H), 2.46(s, 3H), 2.19(m, 1H), 2.00(m, 1H ),1.28(m,2H),0.84(m,4H).

Example 11: (S)-5-(((R)-6-(2-chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3, Preparation of 6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid

Compound 6-(bromomethyl)-4-(2-chloro-3-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (80 mg , 0.18mmol) was dissolved in CHCl3 (15ml), added compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (48mg, 0.27mmol), triethylamine (146mg, 1.44 mmol), heated at 45 °C for 1 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA:MeOH=9:1) to obtain a yellow solid (S)-5-(((R)-6-(2-chloro- 3-Fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptyl Alkane-6-carboxylic acid (10 mg, 11%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =506;

¹ H NMR (400 MHz, CD ₃ OD): δppm 8.17 (m, 2H), 7.37 (m, 1H), 7.29 (m, 2H), 6.31 (s, 1H), 4.56 (m, 1H), 3.87 (m , 1H), 3.69(s, 3H), 3.52(m, 1H), 2.90(m, 1H), 2.51(m, 1H), 1.28(m, 2H), 0.79(m, 4H).

Example 12: (R)-Methyl 4-(2-chloro-3-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-aza Preparation of spiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester

Compound 6-(bromomethyl)-4-(2-chloro-3-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (80 mg , 0.18mmol) was dissolved in CHCl3 (15ml), compound (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (92mg, 0.36mmol) was added ), triethylamine (146 mg, 1.44 mmol), heated at 45 °C for 1 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA:MeOH=9:1) to obtain a yellow solid (R)-methyl 4-(2-chloro-3-fluorophenyl) -6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl )-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (30 mg, 29%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =583;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.46(br,1H), 7.99(m,1H), 7.64(m,1H), 7.26(m,1H), 7.15(m,2H), 6.34(s, 1H), 4.88(m, 1H), 3.80(m, 1H), 3.63(s, 3H), 3.02(s, 3H), 2.76(m, 1H), 2.41(m, 1H), 2.23(m, 1H) ),1.24(m,2H),0.79(m,4H).

Example 13: (1R,3S,5R)-2-(((R)-6-(2-chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl) )-3,6-dihydropyrimidin-4-yl)methyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid preparation

Compound 6-(bromomethyl)-4-(2-chloro-3-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (80 mg , 0.18mmol) was dissolved in CHCl3 (5ml), added compound (1R, 3S, 5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid hydrochloride (44mg, 0.27mmol), three Ethylamine (146 mg, 1.44 mmol) was heated at 45 °C and stirred for 5 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=9/1) to obtain a yellow solid (1R, 3S, 5R)-2-((((R) )-6-(2-Chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3,6-dihydropyrimidin-4-yl)methyl)- 2-Azabicyclo[3.1.0]hexane-3-carboxylic acid (15 mg, 17%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =492;

¹ H NMR (400MHz, CDCl ₃ ): δppm 8.15(br,1H), 7.90(m,1H), 7.83(m,1H), 7.51(m,1H), 7.38(m,2H), 7.23(m, 1H), 6.27(s, 1H), 3.65(s, 3H), 3.44(m, 1H), 2.55(m, 1H), 2.22(m, 1H), 1.78(m, 2H), 0.91(m, 1H) ),0.78(m,2H).

Example 14: (1R,3S,5R)-2-(((R)-6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(4-methylthiazole) Preparation of -2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl The ester (80mg, 0.18mmol) was dissolved in CHCl3 (5ml), compound (1R,3S,5R)-2-azabicyclo[3.1.0]hexane-3-carboxylic acid hydrochloride (59mg, 0.36mmol) was added ), triethylamine (146 mg, 1.44 mmol), heated at 45 °C and stirred for 5 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=9/1) to obtain yellow solid A (10 mg, 11%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =506;

¹ H NMR (400MHz, CD ₃ OD): δppm 7.59(s, 1H), 7.51(m, 1H), 7.25(m, 1H), 7.10(m, 1H), 6.14(s, 1H), 3.79(m ,1H),3.64(s,3H),3.36(m,1H),2.61(s,3H),2.55(m,1H),2.22(m,1H),1.78(m,2H),0.91(m, 1H)0.78(m,2H).

Example 15: (R)-Methyl-4-(2-chloro-4-fluorophenyl)-6-(((S)-6-(isopropylcarbamoyl)-5-azaspiro[ 2.4] Preparation of heptane-5-yl)methyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl The ester (80mg, 0.18mmol) was dissolved in CHCl3 (5ml), compound (S)-N-isopropyl-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (197mg, 0.90mmol) was added ), triethylamine (365 mg, 3.6 mmol), heated at 45 °C for 1 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography using EA as a developing solvent to obtain (R)-methyl-4-(2-chloro-4-fluorophenyl)-6- (((S)-6-(isopropylcarbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(4-methylthiazol-2-yl)- Methyl 1,4-dihydropyrimidine-5-carboxylate (20 mg, 24%).

MS(ESI, pos.ion) m/z: 561[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.20(br,1H), 8.02(br,1H), 7.43(s,1H), 7.15(m,1H), 6.95(m,2H), 6.12(s, 1H), 4.39(m, 1H), 4.13-3.95(m, 2H), 3.75(s, 3H), 2.98-2.70(m, 1H), 2.47(s, 3H), 2.22(m, 1H), 1.93 (m,1H),1.22(m,6H),1.12(m,2H),0.74(m,4H).

Example 16: (R)-Methyl-4-(2-chloro-4-fluorophenyl)-6-(((1R,3S,5R)-3-((methylsulfonyl)carbamoyl) - Preparation of methyl 2-azabicyclo[3.1.0]hexane-2-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (180 mg, 0.41mmol) was dissolved in CHCl3 (10ml), compound (1R,3S,5R)-N-(methylsulfonyl)-2-azabicyclo[3.1.0]hexane-3-carboxamide hydrochloride was added (195mg, 0.81mmol), triethylamine (332mg, 3.28mmol), heated at 45°C and stirred for 5h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography using EA as a developing solvent to obtain (R)-methyl-4-(2-chloro-4-fluorophenyl)-6- (((1R,3S,5R)-3-((methylsulfonyl)carbamoyl)-2-azabicyclo[3.1.0]hexane-2-yl)methyl)-2-(thiazole- 2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (37 mg, 16%).

MS(ESI, pos.ion) m/z: 568 [M+H] ⁺ =569;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.70(br,1H), 8.22(m,1H), 7.73(m,1H), 7.34(m,2H), 7.02(m,1H), 6.11(s, 1H), 4.64(m, 1H), 3.66(s, 3H), 3.22(m, 1H), 3.05(s, 3H), 2.48(m, 2H), 1.43(m, 2H), 0.96(m, 1H) ),0.87(m,1H),0.50(m,1H).

Example 17: (S)-5-(((R)-6-(2-Chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(1-methyl-1H-imidazole Preparation of -2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid

The compound 6-(bromomethyl)-4-(2-chloro-4-fluorophenyl)-2-(1-methyl-1H-imidazol-2-yl)-1,4-dihydropyrimidine-5 -Methyl carboxylate (80mg, 0.18mmol) was dissolved in CHCl3 (5ml), compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (140mg, 0.64mmol) was added, Triethylamine (200 mg, 1.98 mmol) was heated at 45 °C for 5 h. After the reaction was completed, the reaction solution was cooled, concentrated, and the residue was separated and purified by thin layer chromatography (EA:MeOH=9:1) to obtain a yellow solid (20 mg, 18%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =503;

1H NMR (400MHz, CD3OD): δppm 7.49(m,1H), 7.42(m,1H), 7.37(m,1H), 7.28(m,1H), 7.11(m,1H), 6.20(s,1H) ,4.22(m,1H),3.90(s,3H),3.79(m,1H),3.74(s,3H),3.56(m,1H),2.55(m,1H),2.22(m,1H), 1.28(m,2H),0.88(m,4H).

Example 18: (R)-Methyl 4-(2-chloro-3-fluorophenyl)-6-(((S)-6-((cyclopropylsulfonyl)carbamoyl)-5-nitrogen Preparation of methyl heterospiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

Compound 6-(bromomethyl)-4-(2-chloro-3-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (80 mg , 0.18mmol) was dissolved in CHCl3 (15ml), compound (S)-N-(cyclopropylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (96mg, 0.36 mmol), triethylamine (146 mg, 1.44 mmol), heated at 45 °C for 1 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA:MeOH=9:1) to obtain a yellow solid (R)-methyl 4-(2-chloro-3-fluorophenyl) -6-(((S)-6-((cyclopropylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thiazole-2- yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (15 mg, 15%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =609;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.57(br,1H), 8.22(m,1H), 7.54(m,1H), 7.24(m,1H), 7.14(m,2H),

6.32(s, 1H), 4.84(m, 1H), 3.81(m, 1H), 3.77(s, 3H), 3.12(m, 1H), 2.80(m, 1H), 2.43(m, 1H), 2.20( m,1H),1.76(m,1H),1.32(m,2H),0.97(m,4H),0.79(m,4H).

Example 19: (1S,2S,5R)-3-(((R)-6-(2-chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl) )-3,6-dihydropyrimidin-4-yl)methyl)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid.

The compound 6-(bromomethyl)-4-(2-chloro-3-fluorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester (70 mg , 0.16mmol) was dissolved in CHCl3 (5ml), the compound (1S,2S,5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid hydrochloride (53mg, 0.32mmol) was added, DIPEA (203 mg, 1.60 mmol) was heated at 50 °C for 3 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA:MeOH=9:1) to obtain a yellow solid (1S, 2S, 5R)-3-(((R)-6-( 2-Chloro-3-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-3-azadi Cyclo[3.1.0]hexane-2-carboxylic acid (10 mg, 13%).

MS(ESI, pos.ion) m/z: [M+H] ⁺ =491;

¹ H NMR (400 MHz, CD ₃ OD): δppm 7.96 (m, 1H), 7.82 (m, 1H), 7.34-7.11 (m, 3H), 6.20 (s, 1H), 4.59 (m, 1H), 4.40 (m,1H),3.98(m,1H),3.61(s,3H),2.33(m,1H),2.0(m,1H),1.81(m,1H),1.34(m,2H),0.93( m, 2H).

Example 20: (R)-Methyl-4-(2-chloro-4-fluorophenyl)-6-(((S)-6-(hydroxymethyl)-5-azaspiro[2.4]heptyl) Preparation of methyl alkane-5-yl)methyl)-2-(thipyridin-2-yl)-1,4-dihydropyrimidine-5-carboxylate

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (330 mg, 0.74mmol) was dissolved in CHCl3 (15ml), compound (S)-5-azaspiro[2.4]heptane-6-ylmethanol hydrochloride (242mg, 1.48mmol), triethylamine (603mg, 5.92mmol) were added ), heated at 50°C and stirred for 3h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA) to obtain a yellow solid (R)-methyl-4-(2-chloro-4-fluorophenyl)-6-(( (S)-6-(Hydroxymethyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thipyridin-2-yl)-1,4-dihydropyrimidine- Methyl 5-carboxylate (45 mg, 12%).

MS(ESI, pos.ion) m/z: 491[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 7.85(m,1H), 7.48(s,1H), 7.31(m,1H), 7.15(m,1H), 6.94(m,1H), 6.18(s, 1H), 4.48(m, 1H), 4.27(m, 1H), 3.84(m, 3H), 3.78(s, 3H), 3.34(m, 1H), 3.18(m, 1H), 2.92(m, 1H) )2.09(m,2H),0.88(m,4H).

Example 21: (R)-Methyl-4-(2-bromo-4-fluorophenyl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-nitrogen Preparation of methyl heterospiro[2.4]heptan-5-yl)methyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

The compound 4-(2-bromo-4-fluorophenyl)-6-(bromomethyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl The ester (500mg, 0.99mmol) was dissolved in CHCl3 (25ml), compound (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (427mg) was added , 1.68 mmol), triethylamine (800 mg, 8 mmol), heated at 45 ° C and stirred for 5 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA) to obtain a yellow solid (R)-methyl-4-(2-bromo-4-fluorophenyl)-6-(( (S)-6-((Methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(4-methylthiazol-2-yl) -1,4-Dihydropyrimidine-5-carboxylate methyl ester (250 mg, 39%).

MS(ESI, pos.ion) m/z: 639,641 [M+H] ⁺ ;

¹ H NMR (400 MHz, CDCl ₃ ): δppm 7.43 (m, 3H), 7.08 (m, 1H), 6.15 (s, 1H), 4.35 (m, 1H), 4.11 (m, 1H), 3.81 (m, 1H), 3.65(s, 3H), 3.53(m, 1H), 3.05(s, 3H), 2.71(s, 3H), 2.62(m, 2H), 2.45(m, 1H), 1.80(m, 1H ),0.73(m,4H).

Example 22: (S)-5-(((R)-6-(2-bromo-4-fluorophenyl)-5-(methoxycarbonyl)-2-(4-methylthiazol-2-yl )-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid preparation

The compound 4-(2-bromo-4-fluorophenyl)-6-(bromomethyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl The ester (500mg, 0.99mmol) was dissolved in CHCl3 (25ml), compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (352mg, 2mmol), triethylamine (800mg) were added , 8mmol), heated at 50°C and stirred for 5h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH(v/v)=19/1) to obtain a yellow solid (S)-5-(((R)-6- (2-Bromo-4-fluorophenyl)-5-(methoxycarbonyl)-2-(4-methylthiazol-2-yl)-3,6-dihydropyrimidin-4-yl)methyl)- 5-Azaspiro[2.4]heptane-6-carboxylic acid (120 mg, 21%).

MS(ESI, pos.ion) m/z: 562,564 [M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 7.77(s,1H), 7.49(m,2H), 7.21(m,1H), 6.25(s,1H), 3.74(m,2H), 3.67(s, 3H), 3.61(m, 2H), 2.87(m, 1H), 2.69(s, 3H), 2.53(m, 1H), 2.02(m, 2H), 0.90-0.64(m, 4H).

Example 23: (1S,2S,5R)-3-(((R)-6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazole-2- (base)-3,6-dihydropyrimidin-4-yl)methyl)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid

The compound 4-(2-chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (100 mg, 0.22mmol) was dissolved in CHCl3 (8ml), compound (1S, 2S, 5R)-3-azabicyclo[3.1.0]hexane-2-carboxylic acid hydrochloride (100mg, 0.61mmol) was added, three Ethylamine (178 mg, 1.76 mmol) was heated at 50 °C and stirred for 3 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH(v/v)=19/1) to obtain a yellow solid (1S, 2S, 5R)-3-(((R )-6-(2-Chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3,6-dihydropyrimidin-4-yl)methyl) -3-Azabicyclo[3.1.0]hexane-2-carboxylic acid (10 mg, 9%).

MS(ESI, pos.ion) m/z: 492[M+H] ⁺ ;

¹ H NMR (400 MHz, CD ₃ OD): δppm 8.17 (m, 1H), 7.25 (m, 2H), 7.12 (m, 2H), 6.25 (s, 1H), 4.63 (m, 1H), 3.98 (m ,1H),3.61(s,3H),2.33(m,1H),2.0(m,1H),1.81(m,1H),1.34(m,2H),0.93(m,2H).

Example 24: (R)-Methyl-4-(2-chloro-4-fluorophenyl)-2-(1-methyl-1H-imidazol-2-yl)-6-(((S)- Preparation of methyl 6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylate

Compound (S)-N-(methylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (36mg, 0.12mmol) was dissolved in CHCl3 (5ml), 4 -(2-Chloro-4-fluorophenyl)-6-(bromomethyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (27 mg , 0.06mmol), triethylamine (49mg, 0.48mmol), 45 ℃ heating reaction 5h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA:MeOH=9:1) to obtain a yellow solid (R)-methyl-4-(2-chloro-4-fluorophenyl) )-2-(1-Methyl-1H-imidazol-2-yl)-6-(((S)-6-((methylsulfonyl)carbamoyl)-5-azaspiro[2.4]heptane Alk-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (8 mg, 23.5%).

MS(ESI, pos.ion) m/z: 580[M+H] ⁺ ;

¹ H NMR (400 MHz, CDCl ₃ ): δppm 7.38 (m, 1H), 7.36 (m, 2H), 7.33 (m, 2H), 6.24 (s, 1H), 4.35 (m, 1H), 4.11 (m, 1H), 3.81(m, 1H), 3.65(s, 3H), 3.53(m, 1H), 3.05(s, 3H), 2.71(s, 3H), 2.62(m, 2H), 2.45(m, 1H) ),1.80(m,1H),0.83(m,4H).

Example 25: (R)-Methyl-4-(2-bromo-4-fluorophenyl)-2-(4-methylthiazol-2-yl)-6-(((S)-6-( Preparation of (phenylsulfonyl)carbamoyl)-5-azaspiro[2.4]hept-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylate methyl ester

Compound (S)-N-(phenylsulfonyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (204mg, 0.64mmol) was dissolved in CHCl3 (10ml), 4 -(2Bromo-4-fluorophenyl)-6-(bromomethyl)-2-(4-methylthiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (200 mg, 0.40 mmol), triethylamine (324 mg, 3.2 mmol), heated at 50 °C for 5 h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH (v/v)=19/1) to obtain a yellow solid (R)-methyl-4-(2-bromo- 4-Fluorophenyl)-2-(4-methylthiazol-2-yl)-6-(((S)-6-((phenylsulfonyl)carbamoyl)-5-azaspiro[2.4 ]hept-5-yl)methyl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (90 mg, 32%).

MS(ESI, pos.ion) m/z: 702,704 [M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 8.90(br,1H), 7.77(m,2H), 7.45(m,1H), 7.40(m,2H), 7.35(s,1H), 7.27(m, 2H), 6.82(m, 1H), 6.08(s, 1H), 4.29(m, 1H), 4.18(m, 1H), 3.78(m, 1H), 3.64(s, 3H), 3.44(m, 1H) ), 2.79(s, 3H), 2.50(m, 1H), 2.44(m, 1H), 2.03(m, 2H), 0.72(m, 4H).

Example 26: (R)-Methyl-4-(2-chloro-4-fluorophenyl)-6-(((S)-6-(methylcarbamoyl)-5-azaspiro[2.4 ]Heptane-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester

Compound (S)-N-(methyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (380mg, 1.99mmol) was dissolved in CHCl3 (15ml), 4-(2 -chloro-4-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (360 mg, 0.81 mmol), N, N-diisopropylethylamine (1.4ml) was heated at 45°C for 1h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA) to obtain a yellow solid (R)-methyl-4-(2-chloro-4-fluorophenyl)-6-(( (S)-6-(Methylcarbamoyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine -Methyl 5-carboxylate (125 mg, 30%).

MS(ESI, pos.ion) m/z: 518[M+H] ⁺ ;

¹ H NMR (400MHz, CDCl ₃ ): δppm 9.13(br,1H), 7.85(m,1H), 7.54(m,2H), 7.17(m,1H), 6.97(m,1H), 6.20(s, 1H), 4.36(m, 1H), 4.03(m, 1H), 3.60(s, 3H), 2.90-2.70(m, 5H), 2.45(m, 2H), 1.92(m, 2H), 0.64(m ,4H).

Example 27: (R)-Methyl 4-(2-chloro-3-fluorophenyl)-6-(((S)-6-(dimethylaminomethyl)-5-azaspiro[2.4 ]Heptane-5-yl)methyl)-2-(thipyridin-2-yl)-1,4-dihydropyrimidine-5-carboxylate methyl ester

Compound (S)-N-(dimethyl)-5-azaspiro[2.4]heptane-6-carboxamide hydrochloride (130 mg, 0.77 mmol) was dissolved in CHCl3 (6 ml), 4- (2-Chloro-3-fluorophenyl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (115 mg, 0.26 mmol), N,N-diisopropylethylamine (0.45ml) was heated at 45°C for 1h. After the reaction was completed, the reaction solution was cooled, washed and concentrated, and the residue was separated and purified by thin layer chromatography (EA) to obtain a yellow solid (R)-methyl 4-(2-chloro-3-fluorophenyl)-6-( ((S)-6-(Dimethylaminomethyl)-5-azaspiro[2.4]heptan-5-yl)methyl)-2-(thipyridin-2-yl)-1,4- Methyl dihydropyrimidine-5-carboxylate (42 mg, 30%).

MS(ESI, pos.ion) m/z: 533[M+H] ⁺ ;

¹ H NMR (400 MHz, CDCl ₃ ): δppm 7.88 (m, 1H), 7.40 (m, 1H), 7.21 (m, 1H), 7.03 (m, 1H), 6.99 (m, 1H), 6.26 (s, 1H), 4.44(m, 1H), 3.96(m, 1H), 3.82(m, 1H), 3.62(s, 3H), 3.07(s, 3H), 3.01(s, 3H), 2.69(m, 1H) ), 2.25(m, 1H), 1.94(m, 2H), 0.62(m, 4H).

Example 28: (S)-5-(((R)-6-(2-bromo-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3 Preparation of ,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid

Compound (S)-5-azaspiro[2.4]heptane-6-carboxylic acid hydrochloride (360mg, 2.03mmol) was dissolved in CHCl3 (20ml), 4-(2-bromo-4-fluorobenzene was added) yl)-6-(bromomethyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylic acid methyl ester (496 mg, 1.01 mmol), triethylamine (816 mg, 8.08 mmol) , 45 ℃ heating reaction 8h. After the reaction was completed, the reaction solution was cooled and concentrated, and the residue was separated and purified by thin layer chromatography (EA/MeOH(v/v)=9/1) to obtain a yellow solid (S)-5-(((R)-6- (2-Bromo-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-5- Azaspiro[2.4]heptane-6-carboxylic acid (150 mg, 22%)

MS(ESI, pos.ion) m/z: 548,550 [M+H] ⁺ ;

¹ H NMR (400MHz, MeOD): δppm 7.96(m,1H), 7.86(m,1H), 7.55(m,1H), 7.50(m,1H), 7.20(m,1H), 6.15(s,1H) ), 4.59(m, 1H), 4.48(m, 1H), 3.73(m, 1H), 3.64(s, 3H), 3.46(m, 1H), 2.69(m, 1H), 2.14(m, 2H) ,0.84(m,4H).

Example 29: (S)-((isopropoxycarbonyl)oxy)methyl 5-((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-(thiazole Preparation of methyl)-2-yl)-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylate

Compound (S)-5-((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine- 4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (1.0 g, 1.9 mmol) was dissolved in acetonitrile (10 mL), potassium carbonate (1.3 g, 9.4 mmol) was added and catalyzed amount of sodium iodide, replaced with nitrogen three times, and stirred at room temperature for 30 minutes. Chloromethyl isopropyl carbonate (1.5 g, 9.8 mmol) was added to the reaction flask and the temperature was raised to 55° C. to react for 5 hours. After the reaction was completed, the reaction solution was cooled, filtered and concentrated, and the residue was separated and purified by column chromatography (n-heptane/ethyl ester (v/v)=4/1) to obtain a yellow solid (S)-((isopropoxy Carbonyl)oxy)methyl5-((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-(thiazol-2-yl)-3,6-dihydropyrimidine- 4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid methyl ester (750 mg, 60.9%).

MS(ESI, pos.ion) m/z: 621[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ ): δppm 9.75(s,1H), 8.15(m,1H), 7.92(m,1H), 7.41(m,2H), 7.18(m,1H), 6.01( s, 1H), 5.75(m, 1H), 5.67(m, 1H), 4.80(m, 1H), 4.30(m, 1H), 4.10(m, 1H), 3.96(m, 1H), 3.50(s ,3H),2.95(m,1H),2.76(m,1H),2.30(m,1H),1.98(m,1H),1.22(m,6H),0.60(m,4H).

Example 30: (S)-(pivaloyloxy)methyl 5-(((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-(thiazol-2-yl) )-3,6-dihydropyrimidin-4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylate methyl ester

Compound (S)-5-((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine- 4-yl)methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (0.2 g, 0.39 mmol) was dissolved in acetonitrile (2 mL), potassium carbonate (0.269 g, 1.95 mmol) was added and catalyzed amount of sodium iodide, replaced with nitrogen three times, and stirred at room temperature for 30 minutes. Chloromethyl pivalate (0.298 g, 1.98 mmol) was added to the reaction flask and the temperature was raised to 55° C. to react for 5 hours. After the reaction was completed, the reaction solution was cooled, filtered and concentrated, and the residue was separated and purified by column chromatography (n-heptane/ethyl ester (v/v)=4/1) to obtain a yellow solid (S)-(pivaloyloxy ) methyl 5-((6-(2-chloro-4-fluorophenyl)-5-(methoxycarbonyl)-(thiazol-2-yl)-3,6-dihydropyrimidin-4-yl) Methyl)-5-azaspiro[2.4]heptane-6-carboxylic acid methyl ester (180 mg, 73.4%).

MS(ESI, pos.ion) m/z: 619[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ ): δppm 9.84(s,1H), 8.00(m,1H), 7.94(m,1H), 7.43(m,2H), 7.20(m,1H), 6.03( s,1H),5.82(m,1H),5.64(m,1H),,4.31(m,1H),4.11(m,1H),3.91(m,1H),3.61(s,3H),2.95( m, 1H), 2.75(m, 1H), 2.36(m, 1H), 1.98(m, 1H), 1.10(s, 9H), 0.62(m, 4H).

Pharmacodynamic testing experiment

In vitro anti-HBV pharmacodynamic activity test with HBV HepG2.2.15 cell line

1. Experimental method:

qPCR was used to detect the viral DNA content of cell culture medium and calculate the concentration (EC50) when the compound inhibited the virus by half. The specific experimental methods are as follows:

Inoculate HepG2.2.15 cells into a 24-well cell culture plate (200,000 cells/well), and the next day, add cell culture solutions containing different concentrations of the compounds to be tested to treat the cells (the highest concentration of compounds is 5 μM, 5-fold serial dilution, 6 cells) dilution point). On the fifth day, the culture medium containing the drug to be tested was replaced, and on the eighth day, the culture supernatant was collected and centrifuged.

Quantitative PCR: refer to the Hepatitis B Virus Nucleic Acid Quantitative Assay Kit (PCR-Fluorescent Probe Method). Add nucleic acid release agent to the PCR reaction tube, and then add the diluted standard template to each tube (the maximum concentration of the standard template is 4X107IU/mL, ten-fold dilution is 4 points, and the minimum concentration is 4X104IU/mL); add Sample template; configure the reaction mixture according to the PCR system, add it to the reaction tube; cover the PCR reaction tube cap; run the quantitative PCR instrument according to the set program.

Calculation of the percentage inhibition of HBV replication by compounds: %Inh.=[1-total amount of HBV DNA treated with compound/total amount of HBV DNA treated with control]×100.

Calculate the EC ₅₀ value of the compound on HBV replication: use the GraphPad Prism5 analysis software, and use the "four-parameter logistic equation" to calculate the EC ₅₀ value.

2. Experimental results: see Table 1:

Table 1: Anti-HBV Activity of Compounds in HBV HepG2.2.15 Cell Line

Activity interval: a (0.001<a≤0.20μmol).

The experimental results show that the compound of the present invention has a strong anti-HBV virus effect, so it is suitable for treating various diseases caused by HBV virus infection.

Toxicity test

In Vitro Toxicity CC ₅₀ Detection Experiment Using HepG2 Cell Line

1. Experimental method:

The luminescence method cell viability detection kit detects the HepG2 cell viability and calculates the concentration (CC ₅₀ ) when the compound inhibits the HepG2 cell viability by half. The specific experimental method is as follows:

HepG2 cells were seeded into a 96-well cell culture plate (4,000 cells/well), and cell culture medium containing different concentrations of the compounds to be tested was added on the next day to process the cells (the highest compound concentration was 200 μM, 10-fold gradient dilution, 6 dilution points). ). On the fifth day, the cell viability was detected with a luminescence cell viability detection kit.

Calculation of percentage inhibition of HepG2 cell viability by compound: %Inh.=[1-add compound-treated HepG2 cell viability/control-treated HepG2 cell viability]×100.

Calculate the CC ₅₀ value of the compound on the viability of HepG2 cells: the GraphPad Prism5 analysis software was used to calculate the CC ₅₀ value using the "four-parameter logistic equation".

2. Experimental results: see Table 2:

Table 2: _CC50 values of compounds on HepG2 cells

compound CC₅₀(μmol) control compound 82 Example 1 83 Example 2 120 Example 4 132 Example 7 90 Example 8 110 Example 10 115 Example 19 108 Example 22 70 Example 23 96 Example 28 79

Remarks: The structural formula of the reference compound is shown in the following formula, and the preparation method thereof is shown in Example 2 of WO2014037480.

The results of the toxicity test show that the compounds of the present invention are less toxic. Some compounds, eg, the compound of Example 4 (murine _LD50 >1000 mg), showed less toxicity and a better safety profile than the control compound (murine _LD50 >600 mg).

Pharmacokinetic Testing Experiment

Pharmacokinetic Study of Test Compounds in Beagle Dogs

1. Experimental method 1:

Beagle dogs were intravenously injected with 2 mg/kg of the test compound via the forelimb.

Blood was collected from the jugular vein at time points (0.083, 0.25, 0.5, 1, 2, 4, 8, 12 and 24 hours) after administration and collected in anticoagulant tubes with EDTA-K2. Blood samples were centrifuged at 5500 rpm for 10 min to obtain plasma. After liquid-liquid extraction, the plasma samples were quantitatively analyzed by multiple reactive ion monitoring (MRM) on LC-MS/MS. Pharmacokinetic parameters were calculated by non-compartmental model method using WinNonlin6.3 software.

2. Data: see Table 3:

Table 3: Pharmacokinetic data of compounds in Beagle dogs

Pharmacokinetic Testing Experiment

Pharmacokinetic Study of Test Compounds in Beagle Dogs

1. Experimental method 2:

Beagle dogs were orally administered 10 mg/kg of test compound.

Blood was collected from the jugular vein at time points (0.25, 0.5, 1, 2, 4, 8, 12 and 24 hours) after administration, and collected in an anticoagulant tube supplemented with EDTA-K2. Blood samples were centrifuged at 5500 rpm for 10 min to obtain plasma. After liquid-liquid extraction, the plasma samples were quantitatively analyzed by multiple reactive ion monitoring (MRM) on LC-MS/MS. Pharmacokinetic parameters were calculated by non-compartmental model method using WinNonlin6.3 software.

2. Data: see Table 4:

Table 4: Pharmacokinetic data of compounds in Beagle dogs

The results in Table 3 and Table 4 show that: after IV or PO administration to Beagle dogs, compared with the control compound, most of the indicators of the compound of Example 4 are the area under the curve (AUC _0-t ) and the area under the curve (AUC ) _0-inf ), bioavailability (F%) clearance rate (CL) and steady state performance fraction volume (Vss) were much better than the area under the drug-time curve (AUC _0-t ) of the control compound, the drug-time curve Lower area (AUC _0-inf ), bioavailability (F%) clearance (CL) and steady state performance fraction volume (Vss).

In conclusion, the compounds of the present invention have strong anti-HBV effects; and the compounds of the present invention have more favorable pharmacokinetic test results and good toxicity test results, which will make them more likely to be effective and safe drug. In particular, the compound of Example 4 has a moderate clearance rate (4.99mL/kg/min), good bioavailability (48%), and a good toxicity test result (CC ₅₀ is 132 μmol), which indicates that the compound of Example 4 has a good bioavailability (48%). The application prospect of anti-HBV virus is very good.

Claims (13)

1. A compound represented by formula (I), or a pharmaceutically acceptable salt thereof:

wherein:

R₁is phenyl, wherein said phenyl is optionally further substituted with one or more substituents selected from halogen;

R₂is selected from C_1-4An alkyl group;

a is-O-or-S-;

R₃selected from thiazolyl, imidazolyl or pyridyl, said thiaThe azolyl, imidazolyl or pyridyl group may be further substituted with one or more groups selected from halogen or C_1-4Substituted by a substituent of an alkyl group;

r is a group shown as follows:

R₆each independently is- (CR)₇R_7a)_m-OH、-(CR₇R_7a)_n-C(＝O)-NH-S(O)₂-R₉、-(CR₇R_7a)_n-C(＝O)-NR₁₀R_10a、-(CR₇R_7a)_n-C(＝O)O-(CR₇R_7a)_n-OC(＝O)O-R₈、-(CR₇R_7a)_n-C(＝O)O-(CR₇R_7a)_n-OC(＝O)-R₈Or- (CR)₇R_7a)_n-C(＝O)N(R₈)₂；

R₇And R_7aEach independently is hydrogen or C_1-4Alkyl, wherein said C_1-4Alkyl is further substituted with one or more substituents selected from ═ O;

R₈each independently is hydrogen or C_1-4An alkyl group; r₉Is C_1-4Alkyl radical, C_6-10Aryl or C_3-6A cycloalkyl group;

R₁₀and R_10aEach independently is hydrogen or C_1-4Alkyl, or R₁₀And R_10aAnd the nitrogen atom to which it is attached, to form a heterocyclic group, wherein the heterocyclic group is a non-aromatic, saturated or partially unsaturated, monocyclic, bicyclic, or tricyclic ring system of 3 to 6 ring atoms, wherein at least one ring atom is selected from the group consisting of nitrogen, sulfur, and oxygen atoms; m is 1;

n is 0 and 1 independently.

2. The compound of claim 1, wherein R₂Is methyl or ethyl.

3. The compound of claim 1 or 2, wherein a is-O-.

4. The compound of claim 1 or 2, wherein R is selected from:

5. the compound of claim 1, wherein:

R₁is 2 halo-substituted phenyl;

R₂is methyl or ethyl;

a is-O-.

6. A compound selected from:

(1)4- (2-chloro-4-fluorophenyl) -6- (((S) -6- ((methylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(2) (S) -5- ((6- (2-chloro-4-fluorophenyl) -5- (ethoxycarbonyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidin-4-yl) methyl) -5-azaspiro [2.4] heptane-6-carboxylic acid;

(3)4- (2-chloro-4-fluorophenyl) -6- (((S) -6- (morpholine-4-carbonyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(4) (S) -5- (((R) -6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidin-4-yl) methyl) -5-azaspiro [2.4] heptane-6-carboxylic acid;

(5)4- (2-chloro-4-fluorophenyl) -2- (3, 5-fluoropyridin-2-yl) -6- (((S) -6- ((methylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(6)4- (2-chloro-4-fluorophenyl) -2- (3, 5-fluoropyridin-2-yl) -6- ((S) -5-azaspiro [2.4] heptane-6-carboxylic acid) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(7)4- (2-chloro-4-fluorophenyl) -6- (((S) -6- ((methylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid ethyl ester;

(8)4- (2-chloro-4-fluorophenyl) -6- (((S) -6- ((cyclopropylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid ethyl ester;

(9)4- (2-chloro-4-fluorophenyl) -6- (((S) -6- (isopropylcarbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(10) (S) -5- (((R) -6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) -2- (4-methylthiazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) -5-azaspiro [2.4] heptane-6-carboxylic acid;

(11) (S) -5- (((R) -6- (2-chloro-3-fluorophenyl) -5- (methoxycarbonyl) -2- (thiazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) -5-azaspiro [2.4] heptane-6-carboxylic acid;

(12) (R) -methyl 4- (2-chloro-3-fluorophenyl) -6- (((S) -6- ((methylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(15) (R) -methyl-4- (2-chloro-4-fluorophenyl) -6- (((S) -6- (isopropylcarbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (4-methylthiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(17) (S) -5- (((R) -6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) -2- (1-methyl-1H-imidazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) -5-azaspiro [2.4] heptane-6-carboxylic acid;

(18) (R) -methyl 4- (2-chloro-3-fluorophenyl) -6- (((S) -6- ((cyclopropylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(20) (R) -methyl-4- (2-chloro-4-fluorophenyl) -6- (((S) -6- (hydroxymethyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiapyridin-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(21) (R) -methyl-4- (2-bromo-4-fluorophenyl) -6- (((S) -6- ((methylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (4-methylthiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(24) (R) -methyl-4- (2-chloro-4-fluorophenyl) -2- (1-methyl-1H-imidazol-2-yl) -6- (((S) -6- ((methylsulfonyl) carbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(25) (R) -methyl-4- (2-bromo-4-fluorophenyl) -2- (4-methylthiazol-2-yl) -6- (((S) -6- ((phenylsulfonyl) carbamoyl) -5-azaspiro [2.4] hept-5-yl) methyl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(26) (R) -methyl-4- (2-chloro-4-fluorophenyl) -6- (((S) -6- (methylcarbamoyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiazol-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(27) (R) -methyl 4- (2-chloro-3-fluorophenyl) -6- (((S) -6- (dimethylaminomethyl) -5-azaspiro [2.4] heptan-5-yl) methyl) -2- (thiapyridin-2-yl) -1, 4-dihydropyrimidine-5-carboxylic acid methyl ester;

(29) (S) - ((isopropoxycarbonyl) oxy) methyl 5- ((6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) - (thiazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) -5-azaspiro [2.4] heptane-6-carboxylic acid methyl ester;

(30) (S) - (pivaloyloxy) methyl 5- ((6- (2-chloro-4-fluorophenyl) -5- (methoxycarbonyl) - (thiazol-2-yl) -3, 6-dihydropyrimidin-4-yl) methyl) -5-azaspiro [2.4] heptane-6-carboxylic acid methyl ester.

7. A pharmaceutical composition comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 6.

8. Use of a compound according to any one of claims 1 to 6 or a pharmaceutical composition according to claim 7 in the manufacture of a medicament for the treatment or prophylaxis of a disease caused by hepatitis b virus infection.

9. A process for preparing a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1, which process comprises: reacting a compound represented by formula d with an R-H compound or a salt thereof in the presence of a base to obtain a compound represented by formula (I):

wherein R is₁、R₂、R₃R and A are as defined in claim 1.

10. The method of claim 9, wherein the base is an organic base.

11. The production method according to claim 9, wherein the base is triethylamine.

12. The method of claim 9, wherein the compound of formula d is prepared by a method comprising:

step (1): reacting the compound shown in the formula a with the compound shown in the formula b to obtain a compound shown in the formula c:

step (2): reacting the compound shown in the formula c with a brominating reagent to obtain a compound shown in a formula d:

wherein R is₁、R₂、R₃R and A are as defined in claim 1.

13. The method according to claim 9, wherein the brominating reagent in step (2) is N-bromosuccinimide.