WO2020015669A1 - Anti-influenza virus tricyclic derivative - Google Patents

Abstract

Provided are a class of anti-influenza virus compounds and the use of same in the preparation of drugs for treating diseases related to influenza viruses. Specifically, provided are a compound as shown in formula (II) and a pharmaceutically acceptable salt thereof.

Description

Anti-influenza virus tricyclic derivative

References to related applications

This application claims the following priority:

CN201810785083.8, the application date is July 17, 2018;

CN201811416150.5, the application date is November 26, 2018.

Technical field

The invention relates to a class of anti-influenza virus compounds and its application in the preparation of a medicament for treating diseases associated with influenza viruses, and in particular to a compound represented by formula (I), a pharmaceutically acceptable salt thereof, and an optical isomer thereof.

Background technique

Influenza virus, or influenza virus (IFV), is a segmented single-stranded antisense RNA virus that can cause influenza in humans and animals. Influenza viruses can cause very high morbidity and mortality. In particular, influenza A viruses can also cause global pandemics, such as the "Spanish flu" (H1N1 subtype) from 1918 to 1920, and the "Asian flu from 1957 to 1958." "(H2N2 subtype)," Asian influenza "(H3N2 subtype) from 1968 to 1969," Hong Kong flu "(H1N1 subtype) from 1977 to 1978, and the first H1N1 influenza outbreak in Mexico in March 2009. The pandemic caused thousands of deaths, caused great social panic and increased social instability.

Influenza A virus is a single-negative-stranded RNA virus with eight gene segments encoding eight proteins. The 5 ' and 3 ' ends of the influenza virus genome fragment are highly conserved, and the sequences of the two ends are complementary to form a stem-loop structure, which plays an important role in initiating viral RNA replication. The proteins encoded by the various gene fragments of the virus are different in size and play different roles in the life cycle of the influenza virus. First, the basic functions of several major proteins are described below. The influenza virus HA is a ligand that recognizes the host receptor of the influenza virus, and binds to the virus-specific receptor on the cell surface, and mediates the fusion of the outer membrane of the virus with the intracellular small membrane to release the nucleocapsid into the cytoplasm. The receptor for influenza virus is specific, and the receptor for influenza A virus is sialic glycoprotein. The NA protein of influenza virus can remove the sialic acid on the surface of the virus particle during the replication process, so that the virus particle cannot continue to aggregate on the surface of the host cell, which is conducive to the release of the virion and further infect more host cells.

The role of the M2 protein of the influenza virus: The HA protein of the influenza virus binds to sialic acid, and the influenza virus is endocytosed by the host cell. The acidity and alkalinity in phagocytic vesicles play a vital role in virus shelling. The ion channel of M2 protein on the viral membrane can gradually reduce the pH value of phagocytic vesicles. When the pH value drops to 5.0-6.0, it leads to HA2. The protein undergoes allosteric changes, and the fusion peptide located at the amino terminus of the HA2 protein shifts, which in turn activates the fusion process, causing the virus's double-layer lipid membrane to fuse with the cell membrane, releasing the RNPs inside the virus particle to the host cell plasm. M2 protein is a transmembrane ion channel, which is only found in influenza A virus, and part of it extends to the outer membrane surface of the virus.

The synthesis of influenza virus proteins also utilizes the translation mechanism of host cells, and even viruses can suspend the translation of host proteins and accelerate the synthesis of their own proteins. The polyadenylation of host cell mRNA is accomplished by specific adenylate enzymes. The difference is that the adenylate tail of viral mRNA is formed by the continuous transcription of 5-7 uracils on the negative-stranded vRNA. of. The capping of each messenger RNA (mRNA) of the virus is done in a similar way: PA and PB2 proteins take 5 'capped primers from the host's pre-mRNA transcript and then start viral mRNA synthesis, a process called "cap snatching. " After the polyadenylation process and capping process are completed, the viral mRNA exits the nucleus, enters the cytoplasm, and is translated like the host cell's mRNA. The nuclear export of the viral vRNA fragment is mediated by the viral M1 protein and NS2 protein In fact, when M1 protein can interact with vRNA and NP protein, it also interacts with nuclear export protein NS2; therefore, nuclear export protein NS2 mediates M1-RNP to nucleate into the cytoplasm of host cells as a nuclear protein.

Influenza has direct costs due to lost productivity and related medical resources, as well as indirect costs of preventive measures. In the United States, the cumulative flu causes approximately $ 10 billion in losses each year, and it is estimated that future influenza pandemics can cause hundreds of billions of dollars in direct and indirect costs. The cost of prevention is also very high, and governments around the world have spent billions of dollars preparing and planning for a possible H5N1 bird flu pandemic. The costs are related to the purchase of drugs and vaccines, as well as the development of disaster drills and strategies to improve border control.

Current influenza treatment options include vaccination and chemotherapy and chemoprophylaxis with antiviral drugs. Flu vaccination against influenza is often recommended to high-risk groups, such as children and the elderly, or people with asthma, diabetes, or heart disease, but even flu vaccination does not completely prevent the flu. Each season, vaccines for specific influenza strains are re-prepared, but it is impossible to cover the various virus strains that actively infect humans globally during that season. In addition, due to a certain degree of antigen drift in influenza viruses, if more than one virus infects a single cell, eight separate vRNA fragments in the genome are mixed or reassigned, resulting in rapid genetic changes in the virus. Antigen shifts and enable the virus to infect new host species and quickly overcome protective immunity.

Antiviral drugs can also be used to treat influenza. Among them, neuraminidase inhibitors, such as oseltamivir (Duffet), have obvious effects on influenza A virus, but clinical observations have shown that this type of neuraminidase inhibitors Agents have emerged resistant virus strains. In the field of anti-influenza viruses, a new mechanism of action for anti-influenza virus drugs is urgently needed in the clinic, which can support single-agent use for the treatment of influenza A, or through combination with other anti-influenza virus drugs that have been marketed for use in type A Prevention and treatment of influenza.

Among them, WO2016175224 reports the following compounds and their prodrugs:

Summary of the invention

The present invention provides a compound represented by formula (II), a pharmaceutically acceptable salt thereof, and an optical isomer thereof,

among them,

R _{1 is} selected from O and N (R ₄ );

R _{2 is} selected from H, F, Cl, Br, I, OH and NH ₂ ;

R _{3 is} selected from H, F, Cl, Br, I, OH, and NH ₂ ;

R _{4 is} selected from H, OH and C _1-3 alkoxy, said C _1-3 alkoxy is optionally substituted by 1, 2 or 3 R;

R _{5 is} selected from H and C _1-6 alkyl, said C _1-6 alkyl optionally substituted by 1, 2 or 3 R _a ;

R _{6 is} selected from H and C _1-6 alkyl, said C _1-6 alkyl optionally substituted with 1, 2 or 3 R _a ;

Or R ₅ and R _{6 are} linked together to form a C _3-6 cycloalkyl group together with the carbon atom to which they are linked;

R is independently selected from F, Cl, Br, OH, and NH ₂ ;

R _{a is} independently selected from F, Cl, Br, I, OH, NH ₂ and CN;

Carbon atoms with "*" are chiral carbon atoms and exist in the form of a single enantiomer (R) or (S) or are rich in one enantiomer.

The present invention provides a compound represented by formula (II), a pharmaceutically acceptable salt thereof, and an optical isomer thereof,

among them,

R _{1 is} selected from O and N (R ₄ );

R _{2 is} selected from H, F, Cl, Br, I, OH and NH ₂ ;

R _{3 is} selected from H, F, Cl, Br, I, OH, and NH ₂ ;

R _{4 is} selected from H, OH and C _1-3 alkoxy, said C _1-3 alkoxy is optionally substituted by 1, 2 or 3 R;

R _{5 is} selected from H;

R _{6 is} selected from H;

Or R ₅ and R 6 are linked together to form a C _3-5 cycloalkyl group;

R is selected from F, Cl, Br, OH, and NH ₂ ;

Carbon atoms with "*" are chiral carbon atoms and exist in the form of a single enantiomer (R) or (S) or are rich in one enantiomer.

The present invention provides a compound represented by formula (I), a pharmaceutically acceptable salt thereof, and an optical isomer thereof,

among them,

R _{1 is} selected from O and N (R ₄ );

R _{2 is} selected from H, F, Cl, Br, I, OH and NH ₂ ;

R _{3 is} selected from H, F, Cl, Br, I, OH, and NH ₂ ;

R _{4 is} selected from H, OH and C _1-3 alkoxy, said C _1-3 alkoxy is optionally substituted by 1, 2 or 3 R;

R is selected from F, Cl, Br, OH, and NH ₂ .

所述

任选被1、2或3个R取代，其它变量如本发明所定义。 In some aspects of the present invention, R _{4 is} selected from H, OH, and

Said

It is optionally substituted by 1, 2 or 3 R, other variables are as defined in the present invention.

其它变量如本发明所定义。 In some aspects of the present invention, R _{4 is} selected from H, OH, and

Other variables are as defined in the present invention.

In some aspects of the present invention, R _{1 is} selected from O, N (OH), and N (OCH ₃ ), and other variables are as defined in the present invention.

Some aspects of the present invention, the above-described R ₅ is selected from H and C _1-3 alkyl, said C _1-3 alkyl optionally substituted with 1, 2 or 3 R _a, the other variables are as defined in the present invention.

In some aspects of the present invention, R _{5 is} selected from H and CH ₃ , and other variables are as defined in the present invention.

Some aspects of the present invention, the above-described R ₆ is selected from H and C _1-3 alkyl, said C _1-3 alkyl optionally substituted with 1, 2 or 3 R _a, the other variables are as defined in the present invention.

In some aspects of the present invention, the above R _{6 is} selected from H and CH ₃ , and other variables are as defined in the present invention.

选自

其它变量如本发明所定义。 In some aspects of the present invention, the aforementioned structural unit

From

Other variables are as defined in the present invention.

选自

In some aspects of the present invention, the structural unit described above

From

In some embodiments of the present invention, the compound described above, a pharmaceutically acceptable salt thereof, and an optical isomer thereof are selected from the group consisting of

R ₁ , R ₂ , R ₃ , R ₅ and R _{6 are} as defined in any one of claims 1 to 5;

There are still some solutions of the present invention that can be arbitrarily combined from the above variables.

The present invention provides a compound of the following formula, a pharmaceutically acceptable salt thereof, and an optical isomer thereof,

In some embodiments of the present invention, the above compound, a pharmaceutically acceptable salt thereof, and an optical isomer thereof are selected from the group consisting of

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned compound, a pharmaceutically acceptable salt thereof, an optical isomer thereof, and a pharmaceutically acceptable carrier.

The present invention also provides the use of the above compound, a pharmaceutically acceptable salt thereof, an optical isomer thereof, or the above composition in the preparation of a medicament for treating a disease associated with influenza virus.

Technical effect

The compound of the present invention exhibits a positive effect in a test of inhibiting influenza virus replication at a cellular level

Definition and description

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear without a special definition, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding product or its active ingredient.

The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and / or dosage forms that are within the scope of sound medical judgment and are suitable for use in contact with human and animal tissues Without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit / risk ratio.

The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, prepared from a compound having a specific substituent and a relatively non-toxic acid or base found in the present invention. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting a sufficient amount of a base with a neutral form of such compounds in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc .; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also include salts of amino acids (such as arginine, etc.) , And salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain basic and acidic functional groups, and thus can be converted into any base or acid addition salt.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound containing an acid group or a base by a conventional chemical method. Generally, such salts are prepared by reacting these compounds in the form of a free acid or base with a stoichiometric appropriate base or acid in water or an organic solvent or a mixture of the two.

The compounds of the invention may exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including the (R)-and (S) -enantiomers and mixtures and other mixtures thereof, for example, the mixtures of (R)-and (S) -enantiomers, all of these mixtures All fall within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All these isomers and their mixtures are included in the scope of the present invention.

Unless otherwise stated, the term "optical isomers" refers to stereoisomers in a mirror image relationship to each other.

和楔形虚线键

表示一个立体中心的绝对构型，用直形实线键

和直形虚线键

表示立体中心的相对构型，用波浪线

表示楔形实线键

或楔形虚线键

或用波浪线

表示直形实线键

和直形虚线键

Unless otherwise specified, use a wedge solid line key

And wedge dashed keys

Represents the absolute configuration of a solid center, using straight solid line keys

And straight dashed keys

Represents the relative configuration of the solid center, with wavy lines

Represents a wedge solid line key

Or wedge-shaped dotted key

Or with wavy lines

Represents a straight solid line key

And straight dashed keys

连接，则表示该化合物的(Z)型异构体、(E)型异构体或两种异构体的混合物。例如下式(A)表示该化合物以式(A-1)或式(A-2)的单一异构体形式存在或以式(A-1)和式(A-2)两种异构体的混合物形式存在；下式(B)表示该化合物以式(B-1)或式(B-2)的单一异构体形式存在或以式(B-1)和式(B-2)两种异构体的混合物形式存在。下式(C)表示该化合物以式(C-1)或式(C-2)的单一异构体形式存在或以式(C-1)和式(C-2)两种异构体的混合物形式存在。 Unless otherwise stated, when a double bond structure exists in a compound, such as a carbon-carbon double bond, a carbon-nitrogen double bond, and a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (including the nitrogen atom In the double bond, a lone pair of electrons on the nitrogen atom is regarded as a substituent to which it is connected). If the compound on the double bond in the compound has a wavy line

Linking means the (Z) isomer, (E) isomer, or a mixture of the two isomers of the compound. For example, the following formula (A) indicates that the compound exists as a single isomer of formula (A-1) or formula (A-2) or as two isomers of formula (A-1) and formula (A-2) Exists in the form of a mixture; the following formula (B) represents that the compound exists as a single isomer of the formula (B-1) or (B-2) or in the form of both (B-1) and (B-2) The isomers exist as a mixture. The following formula (C) represents that the compound exists as a single isomer of the formula (C-1) or (C-2) or in the form of the two isomers of the formula (C-1) and the formula (C-2) It exists as a mixture.

Unless otherwise stated, the terms "rich in one isomer", "enriched in isomers", "enriched in one enantiomer" or "enantiomerically enriched" refer to one of the isomers or the The enantiomeric content is less than 100%, and the content of the isomer or enantiomer is 60% or more, or 70% or more, or 80% or more, or 90% or more, or 95% or more, or 96% or more, or 97% or more, or 98% or more, or 99% or more, or 99.5% or more, or 99.6% or more, or 99.7% or more, or 99.8% or more, or more 99.9%.

Optically active (R)-and (S) -isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If an enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, in which the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure The desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with a suitable optically active acid or base, and then a conventional method known in the art Diastereomeric resolution is performed and the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereoisomers is usually accomplished by using chromatography that employs a chiral stationary phase and optionally is combined with chemical derivatization (such as the generation of amino groups from amines) Formate). The compounds of the invention may contain atomic isotopes in unnatural proportions on one or more of the atoms constituting the compound. For example, compounds such as tritium ( ³ H), iodine-125 ( ¹²⁵ I) or C-14 ( ¹⁴ C) can be labeled with radioisotopes. As another example, deuterated drugs can be replaced by heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have reduced side effects and increased drug stability. , Enhance efficacy, extend the biological half-life of drugs and other advantages. Transformations of all isotopic compositions of the compounds of the invention, whether radioactive or not, are included within the scope of the invention. "Optional" or "optionally" refers to events or conditions described later that may, but need not, occur, and that the description includes situations in which the events or conditions occur and situations in which the events or conditions do not occur.

The term "substituted" refers to the replacement of any one or more hydrogen atoms on a specific atom with a substituent, and can include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and the substituted compound is stable of. When the substituent is oxygen (= O), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the kind and number of substituents may be arbitrary on the basis of chemically achievable.

When any variable (such as R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if one group is substituted with 0-2 R, the group may be optionally substituted with at most two R, and R in each case has independent options. In addition, combinations of substituents and / or variants are only permitted if such combinations result in stable compounds.

Unless otherwise specified, the term "C _1-6 alkyl" is used to indicate a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. The C _1-6 alkyl includes C _1-5 , C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ and C ₅ alkyl, etc .; it may Is monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of C _1-6 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl and the like.

Unless otherwise specified, the term "C _1-3 alkyl" is used to indicate a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, and the like; it may be monovalent (such as methyl), divalent (such as methylene), or polyvalent (such as methine). . Example C _{1- 3} alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n- propyl and isopropyl) and the like.

Unless otherwise specified, the term "C _1-3 alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule through one oxygen atom. The C _1-3 alkoxy group includes C _1-2 , C _2-3 , C ₃ and C ₂ alkoxy, and the like. Examples of C _1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise specified, "C _3-6 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic and bicyclic system. The C _3-6 cycloalkyl includes C _3-5 , C _4-5 and C _5-6 cycloalkyl and the like; it may be monovalent, divalent or polyvalent. Examples of C _3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

Unless otherwise specified, "C _3-5 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 5 carbon atoms, which is a monocyclic system, and the C _3-5 cycloalkyl includes C _{3 -4} and C _4-5 cycloalkyl and the like; it may be monovalent, divalent or polyvalent. Examples of C _3-5 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and the like.

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those familiar to those skilled in the art. Equivalent alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.

The solvent used in the present invention is commercially available. The present invention uses the following abbreviations: CAN stands for; TFA stands for trifluoroacetic acid; M stands for mol / L; aq stands for water; HATU stands for O- (7-azabenzotriazol-1-yl) -N, N , N ', N'-tetramethylurea hexafluorophosphate; EDC stands for N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride; m-CPBA stands for 3- Chloroperoxybenzoic acid; eq stands for equivalent, equivalent; CDI stands for carbonyldiimidazole; DCM stands for dichloromethane; PE stands for petroleum ether; DIAD stands for diisopropyl azodicarboxylate; DMF stands for N, N-dimethyl DMSO stands for dimethyl sulfoxide; EtOAc stands for ethyl acetate; EtOH stands for ethanol; MeOH stands for methanol; CBz stands for benzyloxycarbonyl, which is an amine protecting group; BOC stands for tert-butoxycarbonyl, which is an amine protecting group Groups; HOAc stands for acetic acid; NaCNBH ₃ stands for sodium cyanoborohydride; rt stands for room temperature; O / N stands for overnight; THF stands for tetrahydrofuran; Boc ₂ O stands for di-tert-butyl dicarbonate; TFA stands for trifluoroacetic acid; DIPEA Stands for diisopropylethylamine; SOCl ₂ stands for thionyl chloride; CS ₂ stands for carbon disulfide; TsOH stands for p-toluenesulfonic acid; NFSI stands for N-fluoro-N- (benzenesulfonyl) benzenesulfonamide; n-B u ₄ NF stands for tetrabutylammonium fluoride; iPrOH stands for 2-propanol; mp stands for melting point; LDA stands for lithium diisopropylamino.

软件命名，市售化合物采用供应商目录名称。 Compounds follow conventional naming principles in the art or use

Software naming. Commercially available compounds use supplier catalog names.

Unless otherwise specified, the silica gel column purification, the silica gel column chromatography, the preparative high-performance liquid phase, and the supercritical fluid chromatography column used in the present invention all have a volume ratio.

detailed description

The following describes the present invention in detail through examples, but it does not imply any disadvantageous limitation to the present invention. The present invention has been described in detail herein, and its specific embodiments are also disclosed. For those skilled in the art, various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. Will be obvious.

Reference example 1: Clips BB-1 and BB-2

synthetic route:

Step 1: Synthesis of the compound BB-1-2

BB-1-1 (10 g, 58.10 mmol) was dissolved in methanol (100 mL), sulfoxide (13.82 g, 116.19 mmol) was added, and the reaction solution was stirred at 80 ° C for 3 hours. The reaction solution was concentrated to dryness to obtain crude BB-1-2, which was directly used in the next reaction.

Step 2: Synthesis of the compound BB-1-3

Compound BB-1-2 (10 g, 53.72 mmol) was dissolved in dichloroethane (150 mL), and bromosuccinimide (10.52 g, 59.09 mmol) and benzoyl peroxide (390.36 mg, 1.61 mmol) were added. ), The reaction solution was stirred at 80 ° C for 12 hours. The reaction solution was washed with an aqueous solution of sodium hydroxide (0.1M, 200 mL) and saturated brine (200 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude BB-1-3, which was directly used in the next step. reaction.

Step 3: Synthesis of the compound BB-1-4

Compound BB-1-3 (14 g, 36.97 mmol) was dissolved in dichloromethane (100 mL), thiophenol (4.41 g, 40.03 mmol) and DBU (6.19 g, 40.67 mmol) were added, and the reaction solution was stirred at 25 ° C for 12 hour. Water (50 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (50 mL × 2). The organic phases were combined and washed with saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was purified by a silica gel column (petroleum ether: ethyl acetate = 50: 1 to 20: 1) to obtain compound BB-1-4.

Step 4: The synthesis of compound BB-1-5

Compound BB-1-4 (6 g, 20.39 mmol) was dissolved in methanol (50 mL) and water (5 mL), sodium hydroxide (1.63 g, 40.77 mmol) was added, and the reaction solution was stirred at 25 ° C. for 12 hours. The reaction solution was adjusted to pH 5 with dilute hydrochloric acid (2M), and the resulting solution was extracted with ethyl acetate (40 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain crude BB-1-5, which was directly used in the next reaction.

Step 5: Synthesis of compound BB-1-6

The mixture BB-1-5 (1 g, 3.57 mmol) and polyphosphoric acid (10 g, 23.78 mmol) were stirred at 120 ° C for 12 hours. The reaction solution was added to ice water (20 mL) and stirred for 10 minutes, and extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with saturated sodium bicarbonate (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. . The obtained crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 50: 1 to 20: 1, volume ratio) to obtain BB-1-6.

Step 6: Synthesis of compound BB-1

Compound BB-1-6 (3 g, 11.44 mmol) was dissolved in ethanol (60 mL), sodium borohydride (865.49 mg, 22.88 mmol) was added in portions at 0 ° C, and the reaction solution was stirred at 25 ° C for 2 hours. The reaction solution was quenched with dilute hydrochloric acid (1M) at ℃, and then extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. It's BB-1. ¹ H NMR (400MHz, deuterated chloroform) δ 7.46-7.47 (m, 1H), 7.14-7.20 (m, 4H), 7.00-7.14 (m, 1H), 6.09 (s, 1H), 4.66-4.70 ( m, 1H), 4.19-4.22 (m, 1H), 2.79 (s, 1H).

Step 7: Synthesis of compound BB-2

Compound BB-1 (0.3 g, 1.14 mmol) was dissolved in DCM (5 mL), and thionyl chloride (675.22 mg, 5.68 mmol, 411.72 uL) was slowly added dropwise. The reaction solution was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to obtain crude BB-2, which was directly used in the next reaction.

Example 1

Step 1: Synthesis of compound 1-2

To a solution of compound 1-1 (3.35 g, 13.61 mmol) in methanol (8 mL) and tetrahydrofuran (32 mL) under an ice bath, a trimethylsilane diazomethane solution (2M, 13.61 mL, 27.22 mmol) was added dropwise, After the addition was completed, the reaction solution was warmed to 20 ° C and stirred for 1 hour. A saturated citric acid solution (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated sodium bicarbonate solution (100 mL) and saturated brine (100 mL), and anhydrous sulfuric acid. Sodium was dried, filtered, and spin-dried to obtain the crude compound 1-2, which was directly used in the next reaction.

Step 2: Synthesis of compounds 1-3

Compound 1-2 (3.98 g, 15.29 mmol), tert-butyl hydrazineformate (2.02 g, 15.29 mmol) and p-toluenesulfonic acid pyridine salt (3.84 g, 15.29 mmol) were added to N, N-dimethylacetamide (80 mL), the reaction solution was reacted at 60 ° C for 12 hours. The reaction solution was cooled to room temperature, water (200 mL) was added, and extraction was performed with ethyl acetate (100 mL × 3). The organic phases were combined, washed with water (200 mL) and saturated brine (200 mL), dried over anhydrous sodium sulfate, and filtered. Spin dry. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1 to 1: 2) to obtain compound 1-3.

Step 3: Synthesis of compound 1-4

Compound 1-3 (2.7 g, 7.21 mmol), methyl acrylate (1.24 g, 14.42 mmol, 1.30 mL), N, N-diisopropylethylamine (2.80 g, 21.64 mmol, 3.77 mL) were dissolved in Acetonitrile (35 mL) was reacted at 50 ° C for 12 hours. The reaction solution was concentrated and dried, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4: 1 to 1: 2, volume ratio) to obtain compound 1-4.

Step 4: Synthesis of compound 1-5

To a solution of compound 1-4 (1.6 g, 3.47 mmol) in ethyl acetate (20 mL) was added a solution of ethyl acetate in hydrochloric acid (4M, 10 mL), and the reaction solution was stirred at 25 ° C. for 1 hour. The reaction solution was spin-dried to obtain crude 1-5, which was directly used in the next reaction.

Step 5: Synthesis of compound 1-6

Compound 1-5 (1.18 g, 3.27 mmol) and potassium tert-butoxide (955.32 mg, 8.51 mmol) were added to acetonitrile (20 mL), and the reaction solution was stirred at 25 ° C. for 1 hour. Methanol (30 mL) was added, concentrated, and dried. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1 to 0: 1, volume ratio, then dichloromethane: methanol = 10: 1 to 0: 1) to obtain compound 1-6.

Step 6: Synthesis of compound 1-7

At 25 ° C, compound 1-6 (0.485 g, 1.48 mmol), BB-1 (390.42 mg, 1.48 mmol) and 1-propylphosphoric anhydride (1.41 g, 2.22 mmol, 1.32 mL, 50% w / w ) Was dissolved in ethyl acetate (5 mL), then methanesulfonic acid (283.95 mg, 2.95 mmol, 210.33 μL) was added, and the reaction solution was stirred at 70 ° C. for 1 hour. The reaction solution was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1 to 0: 1, then ethyl acetate: methanol = 25: 1 to 10: 1, volume ratio) to obtain compound 1-7.

Step 7: Synthesis of Compound 1

Compound 1-7 (0.460 g, 800.57 μmol), sodium chloride solution (116.96 mg, 2.00 mmol in 0.25 mL of water) were added to dimethyl sulfoxide (7 mL), and the reaction solution was stirred at 110 ° C. for 2 hours. The reaction solution was cooled to room temperature, diluted with water (15 mL), and extracted with ethyl acetate (15 mL × 3). The organic phases were combined, washed with saturated brine (15 mL × 2), dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 3: 1 to 0: 1, then dichloromethane: methanol = 10: 1 to 0: 1) to obtain compound 1-8 and crude compound 1, crude product. Compound 1 was separated by preparative high-performance liquid phase (column: Phenomenex Synergi C18 150 × 25mm × 10μm; mobile phase: [A-water (0.225% TFA), B-ACN]; B%: 35% -65%, 10min), Compound 1 was obtained. MS (ESI) m / z: 427.1 (M + H ⁺ ). ¹ H NMR (400 MHz, deuterated methanol) δ 7.30-7.47 (m, 2H), 7.15–7.26 (m, 1H), 7.09–7.15 (m, 2H), 6.68-6.92 (m, 2H), 5.83 5.93 (m, 1H), 5.56-5.71 (m, 1H), 5.42-5.51 (m, 1H), 4.02-4.18 (m, 1H), 3.76-3.96 (m, 1H), 3.41-3.60 (m, 1H ), 3.01-3.21 (m, 1H), 2.53-2.76 (br s, 1H).

Step 8: Synthesis of compounds 1-8A and 1-8B

Compound 1-8 was detected by supercritical fluid chromatography (column model: Chiralpak AD-3 50 × 4.6mm ID, 3 μm; mobile phase: [A: carbon dioxide, B: 0.05% diethylamine ethanol solution, gradient: B%: 40%]; flow rate: 3mL / min; column temperature: 40 ° C; wavelength: 220nm) was analyzed as a racemic compound, and 1-8A (retention time 0.914min) and 1-8B retention time (1.085min) were separated.

Step 9: Synthesis of compound 1A

A solution of compound 1-8A (0.1 g, 193.59 μmol) and lithium chloride (82.06 mg, 1.94 mmol) in N, N-dimethylacetamide (3 mL) was stirred at 80 ° C. for 12 hours. The reaction solution was diluted with water (10 mL), and extracted with ethyl acetate (15 mL × 3). The organic phases were combined, washed with saturated brine (15 mL × 2), dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was separated and purified by preparative high-performance liquid phase (column model: Phenomenex Synergi C18 150 × 30mm × 4μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; gradient: B%: 36% -66% , Retention time: 10 min) to obtain compound 1A. MS (ESI) m / z: 427.2 (M + H ⁺ ). ¹ H NMR (400MHz, d ₄ -MeOH) δ = 7.40 (d, J = 7.4 Hz, 1H), 7.28-7.37 (m, 1H), 7.16-7.25 (m, 1H), 7.08-7.15 (m, 2H ), 6.74-6.89 (m, 2H), 5.87 (d, J = 7.2 Hz, 1H), 5.64 (d, J = 13.6 Hz, 1H), 5.47 (s, 1H), 4.09 (d, J = 13.8 Hz , 1H), 3.78-3.94 (m, 1H), 3.44-3.56 (m, 1H), 3.20-3.28 (m, 1H), 2.58-2.74 (m, 1H).

Step 10: Synthesis of Compound 1B

Compound 1-8B (0.1 g, 193.59 μmol) and a solution of lithium chloride (82.06 mg, 1.94 mmol) in N, N-dimethylacetamide (3 mL) were stirred and reacted at 80 ° C. for 12 hours. The reaction solution was diluted with water (10 mL), and extracted with ethyl acetate (15 mL × 3). The organic phases were combined, washed with saturated brine (15 mL × 2), dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was separated and purified by preparative high-performance liquid phase (column model: Phenomenex Synergi C18 150 × 30mm × 4μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; gradient: B%: 36% -66% , Retention time: 10 min) to obtain compound 1B. MS (ESI) m / z: 427.2 (M + H ⁺ ). ¹ H NMR (400MHz, d ₄ -MeOH) δ = 7.40 (d, J = 7.4 Hz, 1H), 7.27-7.37 (m, 1H), 7.16–7.26 (m, 1H), 7.08-7.15 (m, 2H ), 6.75-6.89 (m, 2H), 5.87 (d, J = 7.2 Hz, 1H), 5.63 (d, J = 15.6 Hz, 1H), 5.42-5.52 (m, 1H), 4.09 (d, J = 13.8Hz, 1H), 3.83-3.99 (m, 1H), 3.44-3.58 (m, 1H), 3.07-3.25 (m, 1H), 2.55-2.76 (m, 1H).

Example 2

synthetic route:

Step 1: Synthesis of Compound 2

Compound 1 (20 mg, 46.90 μmol, 1 eq), hydroxylamine hydrochloride (16.30 mg, 234.50 μmol, 5 eq) and sodium acetate (19.24 mg, 234.50 μmol, 5 eq) were dissolved in ethanol (2 mL), and the reaction solution was reacted at 70 ° C. 5 hour. The reaction solution was concentrated, spin-dried, and the crude product was separated and purified by preparative high-performance liquid phase (column: Phenomenex Synergi C18 150 × 25 × 10 μm; mobile phase: [A-water (0.1% TFA), B-acetonitrile]; B%: 27% -57%, 10 min) to give compound 2.MS (ESI) m / z: 441.8 (M + H ⁺ ). ¹ H NMR (400 MHz, deuterated methanol) δ 7.66 (d, J = 7.2 Hz, 1 H), 7.30-7.41 (m, 1H), 7.19-7.30 (m, 1H), 7.12-7.16 (m, 2H) , 6.79-6.88 (m, 1H), 6.63-6.75 (m, 1H), 6.35 (d, J = 7.2Hz, 1H), 5.63 (dd, J = 2.2, 14.4Hz, 1H), 5.34 (s, 1H ), 4.14 (d, J = 14.0 Hz, 1H), 3.61-3.70 (m, 1H), 3.46-3.55 (m, 1H), 3.14-3.25 (m, 1H), 2.96-3.07 (m, 1H).

Example 3

synthetic route:

Step 1: Synthesis of Compound 3

Compound 1 (20 mg, 46.90 μmol), hydroxylamine hydrochloride (19.59 mg, 234.50 μmol) and sodium acetate (19.24 mg, 234.50 μmol) were dissolved in ethanol (2 mL). The reaction solution was reacted at 70 ° C for 5 hours. The reaction solution was concentrated, spin-dried, and the crude product was separated and purified by preparative high-performance liquid phase (column: Phenomenex Synergi C18 150 * 25 * 10 μm; mobile phase: [A-water (0.1% TFA), B-acetonitrile]; B%: 35% -58%, 9 min) to obtain compound 3. MS (ESI) m / z: 455.9 (M + H ⁺ ). ¹ H NMR (400MHz, DMSO-d6) δ 7.48-7.57 (m, 1H), 7.36-7.44 (m, 1H), 7.29 (d, J = 7.4Hz, 1H), 7.03-7.25 (m, 2H) , 6.73-6.94 (m, 2H), 5.91 (d, J = 7.4 Hz, 1H), 5.54 (d, J = 14.4 Hz, 1H), 5.35 (s, 1H), 4.14 (s, 3H), 4.10 ( d, J = 13.8 Hz, 1H), 3.40-3.52 (m, 2H), 3.18-3.28 (m, 1H), 2.95-3.11 (m, 1H), 2.64-2.87 (m, 1H).

Example 4

Step 1: Synthesis of Compound 4-1

At room temperature, cesium carbonate (776.41 mg, 2.38 mmol, 2.5 eq) was added to tert-butanol (10 ml), compound 1-8 (500 mg, 953.19 μmol) was added to the reaction solution, and the compound (2 was added after stirring for 15 minutes. -Chloroethyl) dimethylsulfoxide iodide (361.08 mg, 1.43 mmol), continued to stir for 12 hours under the protection of nitrogen, and raised the temperature to 60 ° C for 3 hours to react. After the reaction was completed, the mixture was cooled to room temperature, water (30 ml) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and filtered. Concentrated under reduced pressure. The obtained crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate = 5: 1 to 0: 1) to obtain compound 4-1.MS (ESI) m / z: 543.2 (M + H ⁺ ).

Step 2: Synthesis of compounds 4-1A and 4-1B

Compound 4-1 was detected by a supercritical fluid chromatography column (column model: Chiralcel OD-3 50 × 4.6mm ID, 3 μm; mobile phase: [A: carbon dioxide, B: 0.05% diethylamine methanol solution, gradient: B%: 5% to 40%]; flow rate: 3mL / min; column temperature: 40 ° C; wavelength: 220nm) was analyzed as a racemic compound, and the chiral isomers 4-1A (retention time 2.000min) and 4-1B were separated. (Retention time 2.421min).

Step 3: Synthesis of compound 4A

At 25 ° C, compound 4-1A (49 mg, 78.22 μmol, 86.612% purity) was added to a N, N-dimethylacetamide solution (2 ml), and lithium chloride (33.16 mg, 782.16 μmol, 16.02 μL) was added. ), The reaction solution was heated to 90 ° C., and stirred for 4 hours under the protection of nitrogen. After the reaction was completed, it was cooled to room temperature. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate / tetrahydrofuran (4: 1, 10 mL × 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. . The obtained crude product was separated and purified by preparative high-performance liquid phase (column model: Phenomenex Synergi C18 150 × 25 × 10 μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; gradient: B%: 36% -66 %, 10 min) to isolate compound 4A. MS (ESI) m / z: 453.2 (M + H ⁺ ). ¹ HNMR (400MHz, d ₄ -MeOH) δ 7.53 (d, J = 7.2 Hz, 1H), 7.20-7.32 (m, 2H), 7.09-7.15 (m, 2H), 6.78-6.89 (m, 2H) , 5.89 (d, J = 7.2 Hz, 1H), 5.60-5.66 (m, 1H), 5.52 (s, 1H), 4.19-4.28 (m, 1H), 4.12 (d, J = 14.0 Hz, 1H), 2.98-3.06 (m, 1H), 1.89-1.99 (m, 1H), 1.63-1.71 (m, 1H), 1.08-1.17 (m, 1H), 0.94-1.03 (m, 1H).

Step 4: Synthesis of compound 4B

At 25 ° C, compound 4-1B (71 mg, 116.16 μmol, 88.773% purity) was added to a N, N-dimethylacetamide solution (2 ml), and lithium chloride (49.24 mg, 1.16 mmol, 23.79 μL) was added. ), The reaction solution was heated to 90 ° C., and stirred for 4 hours under the protection of nitrogen. After the reaction was completed, it was cooled to room temperature. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate / tetrahydrofuran (4: 1, 10 mL × 3). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude product was separated and purified by preparative high-performance liquid phase (column model: Phenomenex Synergi C18 150 × 25 × 10 μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; gradient: B%: 36% -66 %, 10 min) to isolate compound 4B. MS (ESI) m / z: 453.2 (M + H ⁺ ). ¹ HNMR (400MHz, d ₄ -MeOH) δ 7.53 (d, J = 7.2 Hz, 1H), 7.19-7.32 (m, 2H), 7.07-7.17 (m, 2H), 6.78–6.88 (m, 2H) , 5.89 (d, J = 7.6 Hz, 1H), 5.58-5.68 (m, 1H), 5.52 (s, 1H), 4.19-4.27 (m, 1H), 4.12 (d, J = 13.8 Hz, 1H), 2.98-3.09 (m, 1H), 1.89-1.99 (m, 1H), 1.63-1.71 (m, 1H), 1.07-1.17 (m, 1H), 0.93-1.03 (m, 1H).

Example 5

Step 1: Synthesis of compound 5-2

Add pyrrolidine (278.03 mg, 3.91 mmol, 326.33 μL) and acetic acid (1.17 g, 19.55 mmol, 1.12 mL) to compound 5-1 (3 g, 13.03 mmol) and cyclobutyl formaldehyde (5.48 g, 65.16 mmol, 7.93, respectively). mL) of a dimethyl sulfoxide (20 mL) solution, and the reaction solution was stirred at room temperature for 4 hr. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1/0 to 1/1) to obtain compound 5-2. MS (ESI) m / z: 314.9 (M + H ⁺ ).

Step 2: Synthesis of compound 5-3

Compound 5-2 (2.5 g, 7.95 mmol) was dissolved in methanol (20 mL), acetic acid (716.40 mg, 11.93 mmol, 682.29 μL) and tert-butyl hydrazinoformate (1.26 g, 9.54 mmol) were added. The reaction solution was Stir at 30 ° C for 2hr. Water (50 mL) was added to the reaction solution, and extraction was performed with ethyl acetate (50 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1/0 to 1/1) to obtain compound 5-3. MS (ESI) m / z: 373.3 (M + H ⁺ -56).

Step 3: Synthesis of compound 5-4

Compound 5-3 (550 mg, 1.28 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (3.08 g, 27.01 mmol, 2 mL) was added, and the reaction solution was stirred at room temperature for 2 hr. The reaction solution was concentrated under reduced pressure to obtain a crude compound 5-4, which was directly used in the next reaction. MS (ESI) m / z: 328.9 (M + H ⁺ ).

Step 4: Synthesis of compound 5-5

Compound 5-4 (400 mg, 1.22 mmol) was dissolved in ethanol (10 mL), acetic acid (525.00 mg, 8.74 mmol, 0.5 mL) was added, and the reaction solution was stirred at 70 ° C for 4 hr. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 10/1) to obtain compound 5-5. MS (ESI) m / z: 311.3 (M + H ⁺ ).

Step 5: Synthesis of compound 5-6

Compound 5-5 (200 mg, 644.44 μmol) was dissolved in tetrahydrofuran (3 mL), and sodium cyanoborohydride (121.49 mg, 1.93 mmol) and p-toluenesulfonic acid monohydrate (122.58 mg, 644.44 μmol) were added. The reaction solution was Stir at room temperature for 2hr. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 10/1) to obtain compound 5-6. MS (ESI) m / z: 313.3 (M + H ⁺ ).

Step 6: Synthesis of compound 5-7

Compound 5-6 (110 mg, 352.16 μmol) and BB-2 (181 mg, 640.17 μmol) were dissolved in dichloromethane (3 mL), and triethylamine (79.97 mg, 790.30 μmol, 110.00 μL) was added. The reaction solution was at 15 Stir at ℃ for 15hr. Water (10 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 20/1) to obtain compound 5-7. MS (ESI) m / z: 559.2 (M + H ⁺ ).

Step 7: Synthesis of compound 5-8

Compound 5-7 (80 mg, 143.21 μmol) was dissolved in DCM (2 mL), Dess Martin reagent (78.96 mg, 186.17 μmol) was added, and the reaction solution was stirred at 20 ° C. for 2 hr. Saturated sodium thiosulfate (10 mL) was added to the reaction solution, and after stirring for 10 min, saturated sodium bicarbonate (10 mL) was added and stirred for 10 min, and the reaction solution became clear. Extraction was performed with dichloromethane (20 mL × 2). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 20/1) to obtain compound 5-8. MS (ESI) m / z: 557.2 (M + H ⁺ ).

Step 8: Synthesis of compounds 5-8A and 5-8B

Compound 5-8 was detected by supercritical fluid chromatography (column model: Chiralcel OD-3 50 × 4.6mm ID, 3μm; mobile phase: [A: carbon dioxide, B: 0.05% diethylamine methanol solution, gradient: B%: 5% to 40%]; flow rate: 3mL / min; column temperature: 40 ° C; wavelength: 220nm) was analyzed as a racemic compound, and chiral isomers 5-8A (retention time 1.924min) and 5-8B were separated. (Retention time 2.505min).

Step 9: Synthesis of compound 5A

Compound 5-8A (16.00 mg, 28.74 μmol,) was dissolved in DMA (1 mL), LiCl (2.44 mg, 57.49 μmol) was added, and the mixture was stirred at 90 ° C. for 2 hr. The reaction solution was separated and purified by preparative high performance liquid phase (column model: Xtimate C18 150 * 40mm * 10μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; gradient: B%: 50% -80% , 8min) to give compound 5A. MS (ESI) m / z: 467.1 (M + H ⁺ ).

¹ H NMR (400 MHz, deuterated methanol) δ ppm 7.35-7.45 (m, 1H), 6.92-7.16 (m, 4H), 6.51-6.76 (m, 2H), 5.60-5.82 (m, 1H), 5.44-5.58 (m, 1H), 4.93 (s, 1H), 4.03 (br d, J = 13.55Hz, 1H), 3.56 (s, 2H), 2.76-2.95 (m, 1H), 2.31 (br s, 2H) 2.02 -2.18 (m, 1H) 1.84-2.03 (m, 2H).

Step 10: Synthesis of Compound 5B

Compound 5-8B (16.00 mg, 28.74 μmol) was dissolved in DMA (1 mL), LiCl (2.44 mg, 57.49 μmol) was added, and the mixture was stirred at 90 ° C. for 2 hr. The reaction solution was separated and purified by preparative high performance liquid phase (column model: Xtimate C18 150 * 40mm * 10μm; mobile phase: [A: water (0.225% formic acid), B: acetonitrile]; gradient: B%: 50% -80% , 8min) to obtain compound 5B. MS (ESI) m / z: 467.1 (M + H ⁺ ). ¹ H NMR (400 MHz, deuterated methanol) δ ppm 7.46-7.48 (m, 1H), 7.05-7.16 (m, 4H), 6.74 (br s, 2H), 5.79-5.80 (m, 1H), 5.59-5.63 ( m, 1H), 5.01 (s, 1H), 4.09 (br d, J = 13.80Hz, 1H), 3.74 (s, 2H), 2.93 (br s, 1H), 2.38 (br s, 2H) 2.15-2.28 (m, 1H) 1.84-2.06 (m, 2H).

Example 6

Step 1: Synthesis of compound 6-2

Compound 6-1 (4 g, 17.38 mmol) was dissolved in N, N-dimethylformamide (30 mL) and water (30 mL), and cyclovaleraldehyde (2.05 g, 20.85 mmol) and 2-amino-3- Potassium phenyl-propionate (2.01 g, 12.16 mmol), and the reaction solution was stirred at 20 ° C for 5 hours. Water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by a flash silica gel column (petroleum ether / ethyl acetate = 2/1 to 1/2) to obtain compound 6-2. MS (ESI) m / z: 328.9 (M + H ⁺ ).

Step 2: Synthesis of compound 6-3

Compound 6-2 (2 g, 6.09 mmol) was dissolved in methanol (20 mL), and tert-butyl hydrazinoformate (1.05 g, 7.92 mmol) and acetic acid (182.89 mg, 3.05 mmol, 174.18 μL) were added. The reaction solution was at 20 Stir at 3 ° C for 3 hours. Water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by a flash silica gel column (petroleum ether / ethyl acetate = 2/1 to 1/2) to obtain compound 6-3. MS (ESI) m / z: 443.21 (M + H ⁺ ).

Step 3: Synthesis of compound 6-4

Compound 6-3 (1.2 g, 2.71 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (1.54 g, 13.51 mmol, 1 mL) was added, and the reaction solution was stirred at 20 ° C for 3 hr. The reaction solution was concentrated under reduced pressure to obtain a crude compound 6-4, which was directly used in the next reaction. MS (ESI) m / z: 343.1 (M + H ⁺ ).

Step 4: Synthesis of compound 6-5

Compound 6-4 (0.9 g, 2.63 mmol) was dissolved in ethanol (10 mL), acetic acid (525.00 mg, 8.74 mmol, 0.5 mL) was added, and the reaction solution was stirred at 80 ° C for 1.5 hr. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 10/1) to obtain compound 6-5. MS (ESI) m / z: 325.3 (M + H ⁺ ).

Step 5: Synthesis of compound 6-6

Compound 6-5 (0.2 g, 616.57 μmol) was dissolved in THF (5 mL). Sodium cyanoborohydride (77.49 mg, 1.23 mmol) and p-toluenesulfonic acid monohydrate (117.28 mg, 616.57 μmol) were added. The reaction solution was Stir at 20 ° C for 1 hr. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by a thin-layer chromatography preparation plate (dichloromethane / methanol = 10/1) to obtain compound 6-6. MS (ESI) m / z: 327.0 (M + H ⁺ ).

Step 6: Synthesis of compound 6-7

Compound 6-6 (0.1 g, 306.38 μmol) was dissolved in dichloromethane (3 mL), and BB-2 (129.94 mg, 459.57 μmol) and triethylamine (93.01 mg, 919.15 μmol, 127.93 μL) were added to the reaction solution. Stir at 20 ° C for 12 hr. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by a thin-layer chromatography preparation plate (dichloromethane / methanol = 10/1) to obtain compound 6-7. MS (ESI) m / z: 573.1 (M + H ⁺ ).

Step 7: Synthesis of compound 6-8

Compound 6-7 (90 mg, 157.16 μmol) was dissolved in dichloromethane (3 mL), Dess Martin reagent (0.1 g, 235.77 μmol, 72.99 μL) was added, and the reaction solution was stirred at 20 ° C. for 2 hr. A saturated sodium thiosulfate solution (5 mL) was added to quench the mixture, and the mixture was neutralized with a saturated sodium bicarbonate solution (5 mL), and extracted with dichloromethane (20 mL × 2). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 15/1) to obtain compound 6 -8. MS (ESI) m / z: 571.1 (M + H ⁺ ).

Step 8: Synthesis of compounds 6-8A and 6-8B

Compound 6-8 was detected by supercritical fluid chromatography (column model: ChiralPak AD-3 150 × 4.6mm ID, 3 μm; mobile phase: [A: carbon dioxide, B: 0.05% diethylamine ethanol solution, gradient: B%: 40%]; flow rate: 2.5mL / min; column temperature: 40 ° C; wavelength: 220nm) was analyzed as a racemic compound, and the chiral isomers 6-8A (retention time 3.784min) and 6-8B (retention Time 6.267min).

Step 9: Synthesis of compound 6A

Compound 6-8A (0.017 g, 26.29 μmol) was dissolved in N, N-dimethylacetamide (2 mL), lithium chloride (11.14 mg, 262.86 μmol) was added, and the reaction solution was stirred at 80 ° C. for 12 hr. The reaction solution was filtered, and the filtrate was separated and purified by preparative high-performance liquid phase (column model: Welch Xtimate C18 150 * 25mm * 5μm; mobile phase: [A: water (0.225% FA), B: ACN]; gradient: B%: 50% -80%, 7 min) purification to obtain compound 6A. MS (ESI) m / z: 481.0 (M + H ⁺ ). ¹ H NMR (400 MHz, deuterated methanol) δ 7.57 (d, J = 7.53 Hz, 1H), 7.19 (br d, J = 5.02 Hz, 2H), 7.10 (br d, J = 2.51 Hz, 2H), 6.81 (br s, 2H), 5.87 (d, J = 7.53Hz, 1H), 5.57-5.69 (m, 1H), 5.20 (s, 1H), 4.12 (d, J = 13.55Hz, 1H), 3.87 ( br d, J = 15.06 Hz, 1H), 3.49 (d, J = 15.06 Hz, 1H), 2.59-2.73 (m, 1H), 1.95-2.10 (m, 2H), 1.54-1.91 (m, 5H).

Step 10: Synthesis of compound 6B

Compound 6-8B (0.017 g, 26.29 μmol) was dissolved in N, N-dimethylacetamide (2 mL), lithium chloride (11.14 mg, 262.86 μmol, 5.38 μL) was added, and the reaction solution was stirred at 80 ° C. for 12 hr. . The reaction solution was filtered, and the filtrate was separated and purified by preparative high-performance liquid phase (column model: Welch Xtimate C18 150 * 25mm * 5μm; mobile phase: [A: water (0.225% FA), B: ACN]; gradient: B%: 50 (% -80%, 7 min) to obtain compound 6B. MS (ESI) m / z: 481.0 (M + H ⁺ ). ¹ H NMR (400MHz, deuterated methanol) δ 7.58 (d, J = 7.28 Hz, 1H), 7.20 (br d, J = 4.52 Hz, 2H), 7.10 (br s, 2H), 6.82 (br s, 2H), 5.87 (d, J = 7.28 Hz, 1H), 5.64 (br d, J = 13.30 Hz, 1H), 5.20 (s, 1H), 4.13 (d, J = 13.80 Hz, 1H), 3.87 (br d, J = 14.81 Hz, 1H), 3.50 (br d, J = 15.06 Hz, 1H), 2.61-2.75 (m, 1H), 1.94-2.11 (m, 2H), 1.53-1.93 (m, 5H).

Example 7

Step 1: Synthesis of compound 7-2

Compound 7-1 (2 g, 8.69 mmol) was dissolved in dimethyl sulfoxide (20 mL), cyclohexyl formaldehyde (1.95 g, 17.38 mmol, 2.09 mL), pyrrolidine (185.36 mg, 2.61 mmol, 217.56 μL) and Acetic acid (782.55 mg, 13.03 mmol, 745.29 μL), and the reaction solution was stirred at 15 ° C. for 1 hour. The reaction solution was diluted with ethyl acetate (100 mL), washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by a flash silica gel column (ethyl acetate / petroleum ether = 1/2 to 1/1) to obtain compound 7-2. MS (ESI) m / z: 343.0 (M + H ⁺ ). ¹ H NMR (400MHz, deuterated chloroform) δ 9.72 (s, 1H), 7.64 (d, J = 5.52 Hz, 1H), 7.34-7.45 (m, 5H), 6.41 (d, J = 5.52 Hz, 1H ), 5.17-5.30 (m, 2H), 4.71 (d, J = 7.03 Hz, 1H), 2.03-2.12 (m, 1H), 2.00 (d, J = 7.28 Hz, 1H), 1.70 (br d, J = 15.06 Hz, 1H), 1.54 (br dd, J = 4.27, 7.78 Hz, 2H), 1.30-1.47 (m, 2H), 1.19-1.29 (m, 2H), 1.12 (ddd, J = 3.76, 10.54, 13.80Hz, 1H).

Step 2: Synthesis of compound 7-3

Compound 7-2 (1.9 g, 5.55 mmol) was dissolved in methanol (40 mL), tert-butyl hydrazinoformate (880.08 mg, 6.66 mmol) and acetic acid (333.24 mg, 5.55 mmol, 317.37 μL) were added. The reaction solution was Stir for 1 hour at 20 ° C. The reaction solution was diluted with ethyl acetate (60 mL), washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by a flash silica gel column (ethyl acetate / petroleum ether = 2/1 to 3/2) to obtain compound 7-3. MS (ESI) m / z: 479.1 (M + Na ⁺ ). ¹ H NMR (400 MHz, deuterated chloroform) δ 7.58-7.69 (m, 2H), 7.31-7.47 (m, 5H), 7.12 (s, 1H), 6.39 (d, J = 5.52 Hz, 1H), 5.22 -5.29 (m, 1H), 5.11-5.20 (m, 1H), 4.69 (br d, J = 5.77Hz, 1H), 2.08-2.21 (m, 1H), 1.52-1.58 (m, 4H), 1.51 ( s, 9H), 1.30-1.46 (m, 3H), 1.15-1.30 (m, 2H). Step 3: Synthesis of compound 7-4

Compound 7-3 (2.5 g, 5.48 mmol) was dissolved in dichloromethane (50 mL), hydrogen chloride / dioxane (4M, 5 mL, 20 mmol) was added, and the mixture was stirred at 20 ° C for 12 hours. The reaction solution was concentrated to dryness under reduced pressure, dissolved by adding methanol (30 mL), and then concentrated to dryness under reduced pressure to obtain a crude product 7-4, which was directly used in the next reaction. MS (ESI) m / z: 357.0 (M + H ⁺ )

Step 4: Synthesis of compound 7-5

Compound 7-4 (1.9 g, 5.33 mmol) was dissolved in ethanol (80 mL), acetic acid (8 mL) was added, the reaction solution was stirred at 70 ° C. for 12 hours, and concentrated to dryness under reduced pressure to obtain a crude product. The crude product was purified by a flash silica gel column (dichloromethane / methanol = 1/0 to 10/1) to obtain compound 7-5. MS (ESI) m / z: 339.0 (M + H ⁺ )

Step 5: Synthesis of compound 7-6

Compound 7-5 (400 mg, 1.18 mmol) was dissolved in tetrahydrofuran (8 mL), sodium cyanoborohydride (222.84 mg, 3.55 mmol) was added, and then p-methylbenzenesulfonic acid monohydrate (337.27 mg, 1.77) was added in portions. mmol), and the reaction solution was stirred at 20 ° C for 1 hour. The reaction solution was diluted with ethyl acetate (20 mL), washed with saturated brine (10 mL × 2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by a flash silica gel column (dichloromethane / methanol = 1/0 to 9/1) to obtain compound 7-6. MS (ESI) m / z: 341.1 (M + H ⁺ )

Step 6: Synthesis of compound 7-7

Compound 7-6 (140 mg, 411.26 μmol) was dissolved in dichloromethane (5 mL), and BB-2 (174.42 mg, 616.89 μmol) and triethylamine (83.23 mg, 822.52 μmol, 114.49 μL) were added. Stir at 20 ° C for 12 hours. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by a flash silica gel column (dichloromethane / methanol = 1/0 to 9/1) to obtain a compound 7-7.MS (ESI) m / z: 587.1 (M + H ⁺ ) .

Step 7: Synthesis of compound 7-8

Compound 7-7 (75 mg, 127.84 μmol) was dissolved in dichloromethane (2 mL), Dess-Martin reagent (108.44 mg, 255.67 μmol, 79.15 μL) was added, and the reaction solution was stirred at 20 ° C. for 2 hours. Add saturated sodium thiosulfate solution (5mL), saturated sodium bicarbonate solution (5mL), stir for 3 minutes, extract with dichloromethane (5mL × 3), combine the organic phases, wash with saturated brine (5mL), anhydrous sulfuric acid Dry with sodium, filter, and concentrate the filtrate to dryness to give a crude product. The obtained crude product was purified by a flash silica gel column (dichloromethane / methanol = 1/0 to 9/1) to obtain compound 7-8.MS (ESI) m / z: 585.1 (M + H ⁺ ).

Step 8: Synthesis of compounds 7-8A and 7-8B

Compound 7-8 was detected by a supercritical fluid chromatography column (column model: DAICELCHIRALPAKAD (250mm * 30mm, 10μm); mobile phase: [A: carbon dioxide, B: 0.1% ammonia water ethanol solution, gradient: B%: 55%] ) As a racemic compound, the chiral isomers 7-8A (retention time 0.644min) and 7-8B (retention time 0.991min) were isolated.

Step 9: Synthesis of compound 7A

Compound 7-8A (30.00 mg, 51.31 μmol) was dissolved in N, N-dimethylacetamide (1 mL), lithium chloride (6.53 mg, 153.93 μmol, 3.15 μL) was added, and the reaction solution was stirred at 80 ° C. 12 hours. Take 0.3mL of the reaction solution, dilute with acetonitrile (1mL), filter to obtain the filtrate, and isolate and purify by preparative high-performance liquid phase (column model: Welch Xtimate C18 150 * 25mm * 5μm; mobile phase: [A: water (0.225% formic acid) ; B: acetonitrile]; gradient: B%: 44% -74%, 8.5 min) to give compound 7A. MS (ESI) m / z: 495.3 (M + H ⁺ ). ¹ H NMR (400MHz, deuterated methanol) δ 7.53 (br d, J = 7.53 Hz, 1H), 6.99-7.33 (m, 4H), 6.78 (br s, 2H), 5.87 (br d, J = 7.03 Hz, 1H), 5.67 (br d, J = 14.31 Hz, 1H), 5.12-5.33 (m, 1H), 4.13 (br d, J = 13.80 Hz, 1H), 3.55-3.80 (m, 2H), 2.13 -2.36 (m, 1H), 1.97 (br d, J = 15.31Hz, 1H), 1.41-1.79 (m, 4H), 1.13-1.38 (m, 3H), 0.82-1.09 (m, 1H)

Step 10: Synthesis of compound 7B

Compound 7-8B (30.00 mg, 51.31 μmol) was dissolved in N, N-dimethylacetamide (1 mL), lithium chloride (6.53 mg, 153.93 μmol, 3.15 μL) was added, and the reaction solution was stirred at 80 ° C. 12 hours. Take 0.3mL of the reaction solution, dilute with acetonitrile (1mL), filter to obtain the filtrate, and isolate and purify by preparative high-performance liquid phase (column model: Welch Xtimate C18 150 * 25mm * 5μm; mobile phase: [A: water (0.225% formic acid) B: Acetonitrile]; Gradient: B%: 44% -74%, 8.5 min) to give compound 7B. MS (ESI) m / z: 495.3 (M + H ⁺ ). ¹ H NMR (400 MHz, deuterated methanol) δ 7.54 (d, J = 7.28 Hz, 1 H), 7.18 (br d, J = 4.27 Hz, 2 H), 7.10 (d, J = 3.51 Hz, 2 H), 6.73 -6.88 (m, 2H), 5.87 (d, J = 7.53Hz, 1H), 5.67 (br d, J = 13.80Hz, 1H), 5.24 (s, 1H), 4.13 (d, J = 13.80Hz, 1H ), 3.60-3.76 (m, 2H), 2.25 (br t, J = 13.05 Hz, 1H), 1.98 (br d, J = 14.81 Hz, 1H), 1.49-1.74 (m, 4H), 1.16-1.40 ( m, 3H), 0.96 (br d, J = 11.80Hz, 1H).

Example 8

Step 1: Synthesis of compound 8-2

Compound 8-1 (5 g, 21.72 mmol) and 2-methylpropanal (3.92 g, 54.30 mmol, 4.96 mL) were dissolved in DMSO (50 mL), and pyrrolidine (463.39 mg, 6.52 mmol, 543.89 μL) and acetic acid were added. (1.96 g, 32.58 mmol, 1.86 mL), and the reaction solution was stirred at 20 ° C for 4 hr. Water (50 mL) was added to the reaction solution, and extraction was performed with ethyl acetate (50 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1/0 to 1/1) to obtain compound 3-benzyloxy-2- (1,3-dihydroxy-2,2-dimethyl-3 -Pyrrolidin-1-yl-propyl) pyran-4-one. MS (ESI) m / z: 495.3 (M + H ⁺ ). After the above compound was dissolved in DMSO (20 mL), acetic acid (4.18 g, 69.62 mmol, 3.98 mL) was added, and the reaction solution was stirred at 80 ° C. for 4 hr. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1-1-1 / 2) to obtain compound 8-2. MS (ESI) m / z: 303.0 (M + H ⁺ ).

Step 2: Synthesis of compound 8-3

Compound 8-2 (1 g, 3.31 mmol) was dissolved in methanol (5 mL), and acetic acid (297.94 mg, 4.96 mmol, 283.76 μL) and tert-butyl hydrazineformate (568.30 mg, 4.30 mmol) were added. The reaction solution was at 20 Stir for 2hr. Water (20 mL × 3) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1/0 to 1/1) to obtain compound 8-3. MS (ESI) m / z: 495.3 (M + H ⁺ -56).

Step 3: Synthesis of compound 8-4

Compound 8-4 (660 mg, 1.58 mmol) was dissolved in dichloromethane (6 mL), trifluoroacetic acid (1.85 g, 16.21 mmol, 1.2 mL) was added, and the reaction solution was stirred at 20 ° C for 1 hr. The reaction solution was concentrated under reduced pressure to obtain compound 8-4. MS (ESI) m / z: 316.9 (M + H ⁺ ).

Step 4: Synthesis of compound 8-5

Compound 8-4 (500 mg, 1.58 mmol) was dissolved in EtOH (3.5 mL), acetic acid (681.14 mg, 11.34 mmol, 648.71 μL) was added, and the reaction solution was stirred at 70 ° C. for 16 hr. The reaction solution was concentrated, water (10 mL) was added, and extraction was performed with dichloromethane (10 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (dichloromethane / methanol = 100/1 to 100/4) to purify compound 8-5. MS (ESI) m / z: 298.9 (M + H ⁺ ).

Step 5: Synthesis of compound 8-6

Compound 8-5 (130 mg, 435.75 μmol) was dissolved in tetrahydrofuran (1.5 mL), and sodium cyanoborohydride (82.15 mg, 1.31 mmol) and p-toluenesulfonic acid monohydrate (124.33 mg, 653.63 μmol) were added. Stir for 1 hr. At 20 ° C. Water (10 mL) was added to the reaction solution, and extraction was performed with dichloromethane (10 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography (dichloromethane / methanol = 1/0 to 100/4) to obtain compound 8-6. MS (ESI) m / z: 300.9 (M + H ⁺ ).

Step 6: Synthesis of compound 8-7

Compound 8-6 (100 mg, 332.94 μmol) and BB-2 (188.27 mg, 665.89 μmol) were dissolved in dichloromethane (2 mL), and triethylamine (75.61 mg, 747.18 μmol, 104.00 μL) was added. Stir at 25 ° C for 8hr. Water (20 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 15/1) to obtain compound 8-7.MS (ESI) m / z: 547.9 (M + H ⁺ ).

Step 7: Synthesis of compound 8-8

Compound 8-7 (90 mg, 164.65 μmol) was dissolved in dichloromethane (1 mL), Dess Martin reagent (104.75 mg, 246.97 μmol, 76.46 μL) was added, and the reaction solution was stirred at 20 ° C. for 1 hr. Saturated sodium thiosulfate (5 mL) was added to the reaction solution, and after stirring for 10 min, saturated sodium bicarbonate (5 mL) was added and stirred for 10 min. The reaction solution was clear. Dichloromethane (20 mL × 2) was added for extraction, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin-layer chromatography preparation plate (dichloromethane / methanol = 15/1) to obtain compound 8-8.MS (ESI) m / z: 545.3 (M + H ⁺ ).

Step 8: Synthesis of compounds 8-8A and 8-8B

Compound 8-8 was detected by supercritical fluid chromatography (column model: DAICELCHIRALCELOD-H (250mm * 30mm, 5μm); mobile phase: [A: carbon dioxide, B: 0.1% ammonia water ethanol solution, gradient: B%: 40 %]) Was analyzed as a racemic compound, and the chiral isomers 8-8A (retention time 1.490min) and 8-8B (retention time 1.758min) were isolated.

Step 9: Synthesis of compound 8A

Compound 8-8A (29.35 mg, 53.90 μmol) was dissolved in DMA (1 mL), LiCl (22.85 mg, 538.97 μmol) was added, and the reaction solution was stirred at 90 ° C. for 6 hr. The crude product was separated and purified by preparative high performance liquid phase (column model: Welch Xtimate C18 150 * 25mm * 5μm; mobile phase: [A: water (0.225% formic acid); B: acetonitrile]; gradient: B%: 42% -72% , 8min) was 8A. MS (ESI) m / z: 455.0 (M + H ⁺ ). ¹ H NMR (400 MHz, deuterated chloroform) δ ppm 7.36 (br d, J = 7.28 Hz, 1H), 7.03-7.11 (m, 3H), 6.94 (br s, 1H), 6.80 (br s, 1H), 6.65 (br d, J = 7.53 Hz, 1H), 5.86 (br d, J = 7.28 Hz, 1H), 5.48 (br d, J = 13.30 Hz, 1H), 5.05 (s, 1H), 4.09 (br d, J = 13.30 Hz, 1H), 3.65 (br d, J = 15.31 Hz, 1H), 3.44 (br d, J = 14.81 Hz, 1H), 1.50 (s, 3H), 1.22 (s, 3H). Step 10 :

Synthesis of compound 8B

Compound 8-8B (29.35 mg, 53.90 μmol) was dissolved in DMA (1 mL), LiCl (22.85 mg, 538.97 μmol) was added, and the reaction solution was stirred at 90 ° C. for 6 hr. The crude product was separated and purified by preparative high performance liquid phase (column model: Welch Xtimate C18 150 * 25mm * 5μm; mobile phase: [A: water (0.225% formic acid); B: acetonitrile]; gradient: B%: 42% -72% 8min) to 8B. MS (ESI) m / z: 454.9 (M + H ⁺ ). ¹ H NMR (400 MHz, deuterated chloroform) δ ppm 7.37 (br d, J = 7.53 Hz, 1H), 7.02-7.11 (m, 3H), 6.94 (br s, 1H), 6.81 (br s, 1H), 6.66 (br d, J = 7.28 Hz, 1H), 5.87 (br d, J = 7.53 Hz, 1H), 5.48 (br d, J = 12.80 Hz, 1H), 5.05 (s, 1H), 4.09 (br d, J = 13.55Hz, 1H), 3.65 (br d, J = 14.81Hz, 1H), 3.44 (br d, J = 14.56Hz, 1H), 1.51 (s, 3H), 1.23 (s, 3H).

Biology part

Experimental example 1: Influenza virus cytopathic (CPE) experiment

(EC ₅₀₎ values of the compounds was evaluated antiviral activity against influenza virus (Influenza virus, IFV) measured by the EC50 compound. Cytopathic experiments are widely used to determine the protective effect of compounds on virus-infected cells to reflect the antiviral activity of compounds. Influenza virus CPE experiment

MDCK cells were seeded into a black 384-well cell culture plate at a density of 2,000 cells per well, and then cultured overnight at 37 ° C in a 5% CO ₂ incubator. Compounds were diluted by the Echo555 non-contact nano-upgrade sonic pipette system and added to the cell wells (3-fold dilution, 8 test concentration points). The influenza virus A / Weiss / 43 (H1N1) strain was then added to the cell culture wells at a concentration of 1-2 90% tissue culture infectious dose (TCID90) per well, and the final DMSO concentration in the medium was 0.5%. Set up virus control wells (with DMSO and virus, no compound), cell control wells (with DMSO, no compound and virus), and medium control wells (medium only, no cells). The cytotoxicity and antiviral activity of the compound were measured in parallel. Except that no virus was added, other experimental conditions were consistent with the antiviral activity experiment. The cell plate was cultured in a 5% CO ₂ incubator at 37 ° C for 5 days. After 5 days of incubation, cell viability was detected using the Cell Viability Detection Kit CCK8. Raw data were used for the calculation of the antiviral activity and cytotoxicity of the compounds.

The antiviral activity and cytotoxicity of a compound are represented by the inhibitory rate (%) of the compound on the viral effect of the cell. Calculated as follows:

GraphPad Prism software was used to perform non-linear fitting analysis on the inhibition rate and cytotoxicity of the compounds to obtain the EC ₅₀ values of the compounds. The experimental results are shown in Table 1.

Table 1

Compound EC ₅₀ (μm) Compound EC ₅₀ (μm) 1 0.018 2 1.01 3 0.77 1A 0.157 1B 0.009 4A 2.483 4B 0.002 5A 1.37 5B 0.004 6A > 10 6B 0.007 7A > 1 7B 0.029 8A > 1 8B 0.005 Zh Zh

Results and discussion: The compounds of the present invention have shown a positive effect in the inhibition of influenza virus replication at the cellular level.

Claims (14)

A compound represented by formula (II), a pharmaceutically acceptable salt thereof, and an optical isomer thereof,

among them, R _{1 is} selected from O and N (R ₄ ); R _{2 is} selected from H, F, Cl, Br, I, OH and NH ₂ ; R _{3 is} selected from H, F, Cl, Br, I, OH, and NH ₂ ; R _{4 is} selected from H, OH and C _1-3 alkoxy, said C _1-3 alkoxy is optionally substituted by 1, 2 or 3 R; R _{5 is} selected from H and C _1-6 alkyl, said C _1-6 alkyl optionally substituted by 1, 2 or 3 R _a ; R _{6 is} selected from H and C _1-6 alkyl, said C _1-6 alkyl optionally substituted with 1, 2 or 3 R _a ; Or R ₅ and R _{6 are} linked together to form a C _3-6 cycloalkyl group together with the carbon atom to which they are linked; R is independently selected from F, Cl, Br, OH, and NH ₂ ; R _{a is} independently selected from F, Cl, Br, I, OH, NH ₂ and CN; Carbon atoms with "*" are chiral carbon atoms and exist in the form of a single enantiomer (R) or (S) or are rich in one enantiomer. The compound according to claim 1, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, wherein R _{4 is} selected from the group consisting of H, OH, and

Said

It is optionally substituted with 1, 2 or 3 R. The compound according to claim 2, pharmaceutically acceptable salts and optical isomers thereof, wherein, R ₄ is selected from H, OH and

The compound according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, wherein R _{1 is} selected from the group consisting of O, N (OH), and N (OCH ₃ ). The compound according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, wherein R _{5 is} selected from H and C _1-3 alkyl, and the C _1-3 alkyl It is optionally substituted with 1, 2 or 3 R _a . The compound according to claim 5, pharmaceutically acceptable salts and optical isomers thereof, wherein, R ₅ is selected from H and CH _3. The compound according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, wherein R _{6 is} selected from the group consisting of H and C _1-3 alkyl, and the C _1-3 alkyl It is optionally substituted with 1, 2 or 3 R _a . The compound according to claim 7, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, wherein R _{6 is} selected from H and CH ₃ . The compound according to any one of claims 1 to 3, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, wherein the structural unit

From

The compound according to any one of claims 1 to 8, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, which are selected from the group consisting of

R ₁ , R ₂ , R ₃ , R ₅ and R _{6 are} as defined in any one of claims 1 to 8; A compound of formula, a pharmaceutically acceptable salt thereof, and an optical isomer thereof,

The compound according to claim 11, a pharmaceutically acceptable salt thereof, and an optical isomer thereof, which are selected from the group consisting of

A pharmaceutical composition comprises a therapeutically effective amount of a compound according to any one of claims 1 to 12, a pharmaceutically acceptable salt thereof, an optical isomer thereof, and a pharmaceutically acceptable carrier. Use of a compound according to any one of claims 1 to 12, a pharmaceutically acceptable salt thereof and an optical isomer thereof, or a composition according to claim 13 in the manufacture of a medicament for treating a disease associated with influenza virus.