CN104530030A

CN104530030A - Oxazolidinone compounds and application thereof in drugs

Info

Publication number: CN104530030A
Application number: CN201410758827.9A
Authority: CN
Inventors: 左应林; 郑金付; 文亮; 张瑾; 吴守涛; 袁小凤; 孟禄程; 王晓军; 张英俊
Original assignee: Guangdong HEC Pharmaceutical
Current assignee: Guangdong HEC Pharmaceutical
Priority date: 2014-12-10
Filing date: 2014-12-10
Publication date: 2015-04-22

Abstract

The invention relates to novel oxazolidinone compounds or stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites and pharmaceutically-acceptable salts or prodrugs of the novel oxazolidinone compounds. The invention also relates to application of the novel oxazolidinone compounds or the stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites and pharmaceutically-acceptable salts or prodrugs of the novel oxazolidinone compounds as drugs and particularly relates to application of the novel oxazolidinone compounds or the stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites and pharmaceutically-acceptable salts or prodrugs of the novel oxazolidinone compounds to preparation of drugs for inhibiting factors Xa.

Description

Oxazolidinone compound and application thereof in medicines

Technical Field

The invention belongs to the field of medicaments, and particularly relates to a novel oxazolidinone compound, a pharmaceutical composition and application thereof as a medicament, in particular to application of the oxazolidinone compound as a medicament for preparing a factor Xa inhibitor and application of the oxazolidinone compound as a medicament for treating thromboembolic diseases.

Background

Procoagulant blood (hemostasis) and anticoagulant blood (antithrombosis) are two opposite mechanisms in the human blood system, contradict each other and keep relative balance, and the precise and harmonious process maintains the integrity of the circulatory system. When the function of an anticoagulant fibrinolytic system in vivo is gradually reduced and the blood coagulation and the anticoagulant function in blood are out of balance, the blood coagulation occurs, so that the thrombus or embolism is caused, and the thromboembolic diseases such as myocardial infarction, stroke, deep venous thrombosis, pulmonary embolism and the like are further caused. Thromboembolic disease is the most serious of cardiovascular diseases and is the first killer of human health.

With the further elucidation of the mechanism of thrombosis, many new antithrombotic drugs, namely drugs for inhibiting thrombosis (anticoagulation) and platelet aggregation and drugs for dissolving thrombosis, have been researched and developed aiming at the characteristics and causes of thrombosis. The former has the inhibiting effect on the formation and enlargement of thrombus, and the latter dissolves the formed thrombus to eliminate the harm of thrombotic diseases to human.

The blood coagulation factor Xa is a serine protease which can convert prothrombin into thrombin, is an anticoagulation target with great clinical value, and has important position in controlling thrombin formation and activating coagulation waterfall. Factor Xa, located at the junction of intrinsic and extrinsic coagulation pathways, catalyzes primarily the conversion of factor II to factor IIa, and one inhibitor of factor Xa inhibits the physiological effect of 138 prothrombin molecules due to the amplification of biological signals present during coagulation.

WO 2001047919 discloses a factor Xa inhibitor having a structure which has a high inhibitory effect on both free and bound factor Xa, interrupts the intrinsic and extrinsic pathways of the blood coagulation cascade, and inhibits the generation of thrombin and thrombosis.

It represents the compound 5-chloro-aza- ((5S) -2-oxo-3- [ -4- (3-oxo-4-morpholinyl) phenyl ] -1, 3-oxazolidin-5-yl-2-thiophene-carboxamide marketed as the first factor Xa inhibitor drug at 2008 under the generic name Rivaroxaban or Rivaroxaban.

WO 2003026652 discloses the compound 4,5,6, 7-tetrahydro-1- (4-methoxyphenyl) -7-oxo-6- [4- (2-oxo-1-piperidinyl) phenyl ] -1H-pyrazolo [3,4-c ] pyridine-3-carboxamide having anticoagulant effect, which was marketed in europe in 2011 for the treatment of thromboembolic and acute arterial coronary syndromes, under the generic name Apixaban or Apixaban.

WO 2004058715 discloses the factor Xa inhibitor N' - (5-chloropyridin-2-yl) -N- [ (1S,2R,4S) -4- (dimethylcarbamoyl) -2- [ (5-methyl-6, 7-dihydro-4H- [1,3] thiazolo [5,4-c ] pyridine-2-carbonyl) amino ] cyclohexyl ] ethanediamide, marketed in japan in 2011 under the generic name Edoxaban or Edoxaban, for use in the treatment of venous thromboembolism.

Potent and specific inhibitors of factor Xa are potentially valuable therapeutic agents for the treatment of thromboembolic disorders. The present invention provides a new factor Xa inhibitor-oxazolidinone compound, pharmaceutically acceptable salt or prodrug thereof, which can effectively treat thromboembolic diseases.

Disclosure of Invention

The invention provides oxazolidinone compounds or pharmaceutical compositions thereof, which can effectively treat thromboembolic diseases related to inhibition of factor Xa.

In one aspect, the present invention relates to a compound represented by the general formula (I), or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt, or a prodrug of the compound represented by the formula (I):

wherein,

n is 0 or 1;

R¹is H, -NH₂-CN, F, Cl or Br;

R³is H, -NH₂-CN, F, Cl, Br or C₁-C₄An alkyl group;

x, Y and Z are each independently CR⁴Or N;

each R⁴Independently H, F, Cl, Br, -OH, -C (═ O) OH, -NH₂、C₁-C₄Alkyl or C₁-C₄An alkoxy group; and

R²is H, F, Cl, Br, -OH, -C (═ O) OH, -NH₂、C₁-C₄Alkyl or C₁-C₄An alkoxy group.

In some embodiments of the present invention, the substrate is,

R³is H, -NH₂CN, -F, Cl, Br, methyl, ethyl, n-propyl or isopropyl;

each R⁴Independently H, F, Cl, Br, -OH, -C (═ O) OH, -NH₂Methyl, ethyl, propyl, methoxy, ethoxy or propoxy;

and, R²Is H, F, Cl, Br, -OH, -C (═ O) OH, -NH₂Methyl, ethyl, propyl, methoxy, ethoxy orAnd (6) propoxy.

In some embodiments, the present invention relates to a compound of formula (II), or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, racemate, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of a compound of formula (II):

wherein,

n is 0 or 1;

R¹is H, -NH₂-CN, F, Cl or Br;

x, Y and Z are each independently CR⁴Or N; and

each R⁴Independently H, F, Cl, Br, -OH, -C (═ O) OH, -NH₂Methyl, ethyl, propyl, methoxy, ethoxy or propoxy.

In other embodiments, the invention relates to compounds of one of the following,

or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the above compound.

In another aspect, the invention relates to a pharmaceutical composition comprising a compound of any of the present invention, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or combination thereof.

In another aspect, the invention relates to the use of a compound or pharmaceutical composition according to any of the invention in the manufacture of a medicament for the prevention, treatment or treatment of a thromboembolic disorder.

In some embodiments, the use of the invention, wherein the thromboembolic disorder is myocardial infarction, angina, restenosis following angioplasty or coronary artery bypass, stroke, transient ischemic attack, peripheral arterial occlusive disorder, pulmonary embolism, or deep vein thrombosis.

In some embodiments, the use according to the invention is the use of said compound or said pharmaceutical composition for the manufacture of a medicament for the treatment of Disseminated Intravascular Coagulation (DIC).

In another aspect, the invention relates to the use of a compound according to any one of the invention or a pharmaceutical composition according to the invention in the manufacture of a medicament for inhibiting factor Xa.

The invention encompasses the use of the compounds of the invention and their pharmaceutically acceptable salts for the manufacture of a pharmaceutical product for the treatment of thromboembolic disorders in a patient, including those disorders described herein. The present invention encompasses pharmaceutical compositions comprising therapeutically effective amounts of a compound represented by formula (I) in combination with at least one pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle.

The invention also encompasses a method of treating or ameliorating a thromboembolic disorder, or a predisposition to such a disorder, in a subject, which comprises treating the subject with a therapeutically effective amount of a compound represented by formula (I).

The thromboembolic disorder of the present invention is myocardial infarction, angina pectoris, restenosis following reocclusion and angioplasty or after coronary artery bypass of the aorta, stroke, transient ischemic attack, peripheral arterial occlusive disease, pulmonary embolism or deep vein thrombosis.

Unless otherwise indicated, all stereoisomers, geometric isomers, tautomers, nitroxides, hydrates, solvates, metabolites, salts and pharmaceutically acceptable prodrugs of the compounds of the invention are within the scope of the present invention.

In particular, the salts are pharmaceutically acceptable salts. The term "pharmaceutically acceptable" includes materials or compositions which must be compatible chemically or toxicologically, with the other components comprising the formulation, and with the mammal being treated.

Salts of the compounds of the present invention also include, but are not necessarily pharmaceutically acceptable salts of intermediates used in the preparation or purification of the compounds of formula (I) or isolated enantiomers of the compounds of formula (I).

If the compounds of the invention are basic, the desired salts may be prepared by any suitable method provided in the literature, for example, using inorganic acids such as hydrochloric, hydrobromic, sulfuric, nitric and phosphoric acids and the like. Or using organic acids such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid and salicylic acid; pyranonic acids, such as glucuronic acid and galacturonic acid; alpha-hydroxy acids such as citric acid and tartaric acid; amino acids such as aspartic acid and glutamic acid; aromatic acids such as benzoic acid and cinnamic acid; sulfonic acids such as p-toluenesulfonic acid, ethanesulfonic acid, and the like.

If the compounds of the invention are acidic, the desired salts can be prepared by suitable methods, e.g., using inorganic or organic bases, such as ammonia (primary, secondary, tertiary), alkali or alkaline earth metal hydroxides, and the like. Suitable salts include, but are not limited to, organic salts derived from amino acids such as glycine and arginine, ammonia such as primary, secondary and tertiary amines, and cyclic amines such as piperidine, morpholine, piperazine and the like, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

In another aspect, the invention relates to methods for the preparation, isolation and purification of compounds of formula (I) or (II).

The foregoing merely summarizes certain aspects of the invention and is not intended to be limiting. These and other aspects will be more fully described below.

Detailed Description

Definitions and general terms

Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated by the accompanying structural and chemical formulas. The invention is intended to cover alternatives, modifications and equivalents, which may be included within the scope of the invention as defined by the appended claims. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can 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 documents, patents, and similar materials differ or contradict this application (including but not limited to defined terminology, application of terminology, described techniques, and the like), this application controls.

In the various parts of this specification, substituents of the disclosed compounds are disclosed in terms of group type or range. It is specifically intended that the invention includes each and every independent subcombination of the various members of these groups and ranges. For example, the term "C₁-C₆Alkyl "means in particular independently disclosed methyl, ethyl, C₃Alkyl radical, C₄Alkyl radical, C₅Alkyl and C₆An alkyl group.

The term "alkyl" or "alkyl group" as used herein, denotes a saturated, straight or branched chain monovalent hydrocarbon radical containing from 1 to 20 carbon atoms, wherein the alkyl group may be optionally substituted with one or more substituents as described herein. In some embodiments, the alkyl group contains 1 to 12 carbon atoms; in a further embodiment of the process according to the invention,the alkyl group contains 1 to 6 carbon atoms; in yet another embodiment, the alkyl group contains 1 to 4 carbon atoms; in yet another embodiment, the alkyl group contains 1 to 3 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me, -CH)₃) Ethyl (Et-CH)₂CH₃) N-propyl (n-Pr, -CH)₂CH₂CH₃) Isopropyl (i-Pr, -CH (CH)₃)₂) N-butyl (n-Bu, -CH)₂CH₂CH₂CH₃) Isobutyl (i-Bu, -CH)₂CH(CH₃)₂) Sec-butyl (s-Bu, -CH (CH)₃)CH₂CH₃) Tert-butyl (t-Bu, -C (CH)₃)₃) N-pentyl (-CH)₂CH₂CH₂CH₂CH₃) 2-pentyl (-CH (CH)₃)CH₂CH₂CH₃) 3-pentyl (-CH (CH)₂CH₃)₂) 2-methyl-2-butyl (-C (CH)₃)₂CH₂CH₃) 3-methyl-2-butyl (-CH (CH)₃)CH(CH₃)₂) 3-methyl-1-butyl (-CH)₂CH₂CH(CH₃)₂) 2-methyl-1-butyl (-CH)₂CH(CH₃)CH₂CH₃) N-hexyl (-CH)₂CH₂CH₂CH₂CH₂CH₃) 2-hexyl (-CH (CH)₃)CH₂CH₂CH₂CH₃) 3-hexyl (-CH (CH)₂CH₃)(CH₂CH₂CH₃) 2-methyl-2-pentyl (-C (CH))₃)₂CH₂CH₂CH₃) 3-methyl-2-pentyl (-CH (CH)₃)CH(CH₃)CH₂CH₃) 4-methyl-2-pentyl (-CH (CH)₃)CH₂CH(CH₃)₂) 3-methyl-3-pentyl (-C (CH)₃)(CH₂CH₃)₂) 2-methyl-3-pentyl (-CH (CH)₂CH₃)CH(CH₃)₂) 2, 3-dimethyl-2-butyl (-C (CH)₃)₂CH(CH₃)₂) 3, 3-dimethyl-2-butyl (-CH (CH)₃)C(CH₃)₃) N-heptyl, n-octyl, and the like.

The term "alkoxy" means an alkyl group attached to the rest of the molecule through an oxygen atom, wherein the alkyl group has the meaning as described herein. Unless otherwise specified, the alkoxy group contains 1 to 12 carbon atoms. In one embodiment, the alkoxy group contains 1 to 6 carbon atoms; in another embodiment, the alkoxy group contains 1 to 4 carbon atoms; in yet another embodiment, the alkoxy group contains 1 to 3 carbon atoms. Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH)₃) Ethoxy (EtO, -OCH)₂CH₃) 1-propoxy (n-PrO, n-propoxy, -OCH)₂CH₂CH₃) 2-propoxy (i-PrO, i-propoxy, -OCH (CH)₃)₂) 1-butoxy (n-BuO, n-butoxy, -OCH)₂CH₂CH₂CH₃) 2-methyl-l-propoxy (i-BuO, i-butoxy, -OCH)₂CH(CH₃)₂) 2-butoxy (s-BuO, s-butoxy, -OCH (CH)₃)CH₂CH₃) 2-methyl-2-propoxy (t-BuO, t-butoxy, -OC (CH)₃)₃) 1-pentyloxy (n-pentyloxy, -OCH)₂CH₂CH₂CH₂CH₃) 2-pentyloxy (-OCH (CH)₃)CH₂CH₂CH₃) 3-pentyloxy (-OCH (CH))₂CH₃)₂) 2-methyl-2-butoxy (-OC (CH))₃)₂CH₂CH₃) 3-methyl-2-butoxy (-OCH (CH)₃)CH(CH₃)₂) 3-methyl-l-butoxy (-OCH)₂CH₂CH(CH₃)₂) 2-methyl-l-butoxy (-OCH)₂CH(CH₃)CH₂CH₃) And so on.

Any formulae given herein are also intended to represent the non-isotopically enriched forms as well as the isotopically enriched forms of these compounds. Isotopically enriched compounds have the structure depicted by the formulae given herein, except that one or more atoms are replaced by atoms having the selected atomAtomic substitution of mass or mass numbers. Exemplary isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as²H、³H、¹¹C、¹³C、¹⁴C、¹⁵N、¹⁷O、¹⁸O、¹⁸F、³¹P、³²P、³⁵S、³⁶Cl and¹²⁵I。

in addition, heavier isotopes are, in particular, deuterium (i.e.,²substitution of H or D) may provide certain therapeutic advantages resulting from greater metabolic stability. For example, increased in vivo half-life or decreased dosage requirements or improved therapeutic index. It is to be understood that deuterium in the present invention is considered as a substituent of the compound of formula (I). The concentration of such heavier isotopes, particularly deuterium, can be defined by isotopic enrichment factors. The term "isotopic enrichment factor" as used herein refers to the ratio between the isotopic and natural abundance of a given isotope. If a substituent of a compound of the invention is designated as deuterium, the compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). Pharmaceutically acceptable solvates of the invention include those in which the crystallization solvent may be isotopically substituted, e.g. D₂O, acetone-d₆、DMSO-d₆Those solvates of (a).

The term "thromboembolic disease" as used herein refers to a disease caused by two pathological processes, namely thrombosis and thromboembolism, and is also referred to as thrombotic disease. Thrombosis refers to a pathological process in which blood components form emboli in blood vessels or local parts of the endocardium under certain conditions, so that the blood vessels are partially or completely blocked, and the blood supply of corresponding parts is blocked. Thromboembolism is a pathological process in which a thrombus falls off from a formation part and partially or completely blocks blood vessels in the process of flowing along with blood, causing ischemia, anoxia, necrosis, blood stasis and edema of the blood vessels or the system. Examples of thromboembolic disorders include, but are not limited to, arterial cardiovascular thromboembolic disorders, venous cardiovascular thromboembolic disorders, and thromboembolic disorders in the chambers of the heart. More specific examples of such conditions include, but are not limited to, myocardial infarction, angina (including unstable angina), acute coronary syndrome, restenosis following reocclusion and angioplasty or aortic coronary bypass, stroke, transient ischemic attacks, peripheral arterial occlusive disease, arterial thrombosis, coronary thrombosis, cerebral arterial thrombosis, cerebral embolism, renal embolism, pulmonary embolism, thrombophlebitis, venous thrombosis or deep vein thrombosis, and the like.

The term "subject" as used herein refers to an animal. Typically the animal is a mammal. Subjects, e.g., also primates (e.g., humans, males or females), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, etc. In certain embodiments, the subject is a primate. In other embodiments, the subject is a human.

The term "patient" as used herein refers to a human or other animal. In some embodiments, "patient" refers to a human.

The term "treating" or "treatment" as used herein refers, in some embodiments, to ameliorating a disease or disorder (i.e., slowing or arresting or reducing the development of the disease or at least one clinical symptom thereof). In other embodiments, "treating" or "treatment" refers to moderating or improving at least one physical parameter, including physical parameters that may not be perceived by the patient. In other embodiments, "treating" or "treatment" refers to modulating the disease or disorder, either physically (e.g., stabilizing a perceptible symptom) or physiologically (e.g., stabilizing a parameter of the body), or both. In other embodiments, "treating" or "treatment" refers to preventing or delaying the onset, occurrence, or worsening of a disease or disorder.

"stereoisomers" refers to compounds having the same chemical structure but differing in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric isomers (cis/trans), atropisomers, and the like.

"chiral" is a molecule having the property of not overlapping its mirror image; and "achiral" refers to a molecule that can overlap with its mirror image.

"enantiomer" refers to two isomers of a compound that are not overlapping but are in mirror image relationship to each other.

"diastereomer" refers to a stereoisomer that has two or more chiral neutrals and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may be separated by high resolution analytical procedures such as electrophoresis and chromatography, e.g., HPLC.

The stereochemical definitions and rules used in the present invention generally follow the general definitions of S.P. Parker, Ed., McGraw-Hill Dictionary of chemical terms (1984) McGraw-Hill Book Company, New York; and Eliel, E.and Wilen, S., "Stereochemistry of organic Compounds", John Wiley & Sons, Inc., New York, 1994.

Many organic compounds exist in an optically active form, i.e., they have the ability to rotate the plane of plane polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of a molecule with respect to one or more of its chiral centers. The prefixes d and l or (+) and (-) are the symbols used to specify the rotation of plane polarized light by the compound, where (-) or l indicates that the compound is left-handed. Compounds prefixed with (+) or d are dextrorotatory. A particular stereoisomer is an enantiomer and a mixture of such isomers is referred to as an enantiomeric mixture. A50: 50 mixture of enantiomers is referred to as a racemic mixture or racemate, which may occur when there is no stereoselectivity or stereospecificity in the chemical reaction or process.

Depending on the choice of starting materials and methods, the compounds of the invention may exist as one of the possible isomers or as mixtures thereof, for example as racemates and mixtures of non-corresponding isomers (depending on the number of asymmetric carbon atoms). Optically active (R) -or (S) -isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituents may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl group, the substituents of the cycloalkyl group may have cis or trans configuration.

Any resulting mixture of stereoisomers may be separated into pure or substantially pure geometric isomers, enantiomers, diastereomers, depending on differences in the physicochemical properties of the components, for example, by chromatography and/or fractional crystallization.

The term "tautomer" or "tautomeric form" refers to structural isomers having different energies that can interconvert by a low energy barrier (low energy barrier). If tautomerism is possible (e.g., in solution), then the chemical equilibrium of the tautomer can be reached. For example, proton tautomers (also known as proton transfer tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers (valenctautomers) include interconversion by recombination of some of the bonding electrons. A specific example of keto-enol tautomerism is the tautomerism of the pentan-2, 4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-ketone tautomerism. One specific example of phenol-ketone tautomerism is the tautomerism of pyridin-4-ol and pyridin-4 (1H) -one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention.

The term "prodrug", as used herein, represents a compoundIs converted into the compound shown in the formula (I) in vivo. Such conversion is effected by hydrolysis of the prodrug in the blood or by enzymatic conversion to the parent structure in the blood or tissue. The prodrug compound of the invention can be ester, and in the prior invention, the ester can be used as the prodrug and comprises phenyl ester and aliphatic (C)_1-24) Esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters. For example, a compound of the present invention contains a hydroxy group, i.e., it can be acylated to provide the compound in prodrug form. Other prodrug forms include phosphate esters, such as those obtained by phosphorylation of a hydroxyl group on the parent. For a complete discussion of prodrugs, reference may be made to the following: T.Higuchi and V.Stella, Pro-drugs as in novel Systems, Vol.14of the A.C.S.Symphosium Series, Edward B.Roche, ed., Bioversible arrays in Drug designs, American Pharmaceutical Association and Pergamon Press,1987, J.Rautio et al, Prodrugs: Design and Clinical Applications, Nature Review Discovery,2008,7,255 and 270, and S.J.Herr et al, Prodrugs of pharmaceuticals and pharmaceuticals, Journal of chemical Chemistry,2008,51, 2328 and 2345.

"metabolite" refers to the product of a particular compound or salt thereof obtained by metabolism in vivo. Metabolites of a compound can be identified by techniques well known in the art, and its activity can be characterized by assay methods as described herein. Such products may be obtained by administering the compound by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic cleavage, and the like. Accordingly, the present invention includes metabolites of compounds, including metabolites produced by contacting a compound of the present invention with a mammal for a sufficient period of time.

As used herein, "pharmaceutically acceptable salts" refer to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as are: s.m.berge et al, j.pharmaceuticall Sciences, 66: 1-19,1977. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, salts of inorganic acids formed by reaction with amino groups such as hydrochlorides, hydrobromides, phosphates, sulfates, perchlorates, and salts of organic acids such as acetates, oxalates, maleates, tartrates, citrates, succinates, malonates, or those obtained by other methods described in the literature above, such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, cyclopentylpropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, glucoheptonates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxy-ethanesulfonates, lactobionates, lactates, laurates, malates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, palmitates, pamoates, pectinates, persulfates, 3-phenylpropionates, picrates, pivalates, propionates, stearates, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts obtained with appropriate bases include alkali metals, alkaline earth metals, ammonium and N⁺(C_1-4Alkyl radical)₄A salt. The present invention also contemplates quaternary ammonium salts formed from compounds containing groups of N. Water-soluble or oil-soluble or dispersion products can be obtained by quaternization. Alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts and amine cations resistant to formation of counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C_1-8Sulfonates and aromatic sulfonates.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, basic or acidic moiety, by conventional chemical methods. In general, such salts can be prepared by reacting the free acid forms of these compounds with a stoichiometric amount of the appropriate base (e.g., Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, etc.), or by reacting the free base forms of these compounds with a stoichiometric amount of the appropriate acid. Such reactions are usually carried out in water or an organic solvent or a mixture of both. Generally, where appropriate, it is desirable to use a non-aqueous medium such as diethyl ether, ethyl acetate, ethanol, isopropanol or acetonitrile. In, for example, "Remington's Pharmaceutical Sciences", 20 th edition, Mack publishing company, Easton, Pa., (1985); and "handbook of pharmaceutically acceptable salts: properties, Selection and application (Handbook of Pharmaceutical Salts: Properties, Selection, and Use) ", Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002) may find some additional lists of suitable Salts.

"solvate" of the present invention refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol. The term "hydrate" refers to an association of solvent molecules that is water.

In addition, the compounds disclosed herein, including their salts, may also be obtained in the form of their hydrates or in the form of solvents containing them (e.g., ethanol, DMSO, etc.), for their crystallization. The compounds disclosed herein may form solvates with pharmaceutically acceptable solvents (including water), either inherently or by design; thus, the present invention is intended to include both solvated and unsolvated forms.

The "Curtius rearrangement" is a rearrangement reaction in which the rearrangement of acyl azides to isocyanates is first observed in Curtius (Theodor Curtius). The product can be reacted with a range of nucleophiles: hydrolyzing with water to obtain amine; reacting with benzyl alcohol to produce amines with benzyloxycarbonyl protecting groups (Cbz); reacting with tert-butanol to generate amines with a tert-butoxycarbonyl protecting group (Boc) which are used as important intermediates in organic synthesis.

The term "protecting group" or "PG" refers to a substituent that, when reacted with other functional groups, is generally used to block or protect a particular functionality. For example, "amino protecting group" means a substituent attached to an amino group to block or protect the functionality of the amino group in a compound, and suitable amino protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC ), benzyloxycarbonyl (CBZ ) and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, "hydroxyl protecting group" refers to the functionality of a substituent of a hydroxyl group to block or protect the hydroxyl group, and suitable protecting groups include acetyl and silyl groups. "carboxy protecting group" refers to the functionality of a substituent of a carboxy group to block or protect the carboxy group, and typical carboxy protecting groups include-CH₂CH₂SO₂Ph, cyanoethyl, 2- (trimethylsilyl) ethyl, 2- (trimethylsilyl) ethoxymethyl, 2- (p-toluenesulfonyl) ethyl, 2- (p-nitrobenzenesulfonyl) ethyl, 2- (diphenylphosphino) ethyl, nitroethyl, and the like. General descriptions of protecting groups can be found in the literature: greene, Protective Groups in Organic Synthesis, John Wiley&Sons,New York,1991；and P.J.Kocienski,Protecting Groups,Thieme,Stuttgart,2005。

Compounds of the invention and pharmaceutical compositions, formulations, administration and uses

According to another aspect, a pharmaceutical composition of the invention is characterized by comprising a compound of formula (I), a compound listed in the present invention, or a compound of the examples, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of compound in the compositions of the invention is effective to treat or ameliorate thromboembolic disorders in a patient or as a factor Xa inhibitor.

The compounds of the invention exist in free form or, where appropriate, as pharmaceutically acceptable derivatives. According to the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable prodrugs, salts, esters, salts of esters, or any other adduct or derivative that can be administered directly or indirectly in accordance with the needs of the patient, compounds described in other aspects of the invention, metabolites thereof, or residues thereof.

As described herein, the pharmaceutically acceptable compositions of the present invention further comprise a pharmaceutically acceptable carrier, adjuvant, or excipient, as used herein, including any solvent, diluent, or other liquid excipient, dispersant or suspending agent, surfactant, isotonic agent, thickening agent, emulsifier, preservative, solid binder or lubricant, and the like, as appropriate for the particular target dosage form. As described in the following documents: in Remington, The Science and Practice of Pharmacy,21st edition,2005, ed.D.B.Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds.J.Swarbrick and J.C.Boylan, 1988. Annu 1999, Marcel Dekker, New York, taken together with The disclosure of this document, suggests that different carriers may be employed In The preparation of pharmaceutically acceptable compositions and their well known methods of preparation. Except insofar as any conventional carrier vehicle is incompatible with the compounds of the invention, e.g., any adverse biological effect produced or interaction in a deleterious manner with any other component of a pharmaceutically acceptable composition, its use is contemplated by the present invention.

Substances which may serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-blocking polymers, lanolin, sugars, such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; gum powder; malt; gelatin; talc powder; adjuvants such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic salt; ringer's solution; ethanol, phosphate buffered solutions, and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate, coloring agents, releasing agents, coating materials, sweetening, flavoring and perfuming agents, preservatives and antioxidants.

The compounds of the present invention may be administered in the form of oral dosage forms such as tablets, capsules (each of which includes sustained release or timed release formulations), pills, powders, granules, elixirs, tinctures, suspensions, syrups, and emulsions. They may also be administered intravenously (bolus or infusion), intraperitoneally, subcutaneously, or intramuscularly, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts. They may be administered separately, but will generally be administered together with a pharmaceutical carrier selected based on the mode of administration selected and standard pharmaceutical practice.

The dosage regimen for a compound of the invention will vary depending upon a variety of factors known, such as the pharmacokinetic characteristics of the particular agent and its mode and route of administration; race, age, sex, health condition, medical condition, and weight of the recipient; the nature and extent of the symptoms; the kind of concurrent therapy; the frequency of treatment; the route of administration, the renal and hepatic function of the patient, and the desired effect. A physician or veterinarian can make a decision and prescribe the effective amount of the drug to prevent, counter or arrest the progress of the thromboembolic disorder.

In accordance with general guidelines, the daily oral dosage of each active ingredient used is in the range of about 0.001 to 1000mg/kg body weight, preferably about 0.01 to 100mg/kg body weight, in order to achieve the indicated effect. And, most preferably, between about 1.0 and 20mg/kg body weight/day. For intravenous administration, the most preferred dosage range during infusion at conventional rates is from about 1 to about 10mg/kg body weight/minute. The compounds of the invention may be administered once daily, or may be administered in divided doses of two, three or four times daily.

The compounds of the invention may be administered in intranasal form via topical use of suitable intranasal vehicles, or by the transdermal route using transdermal patches. When administered in the form of a transdermal delivery system, the dosage administered throughout the administration period is continuous rather than intermittent.

The compounds of the invention may also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from different phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.

The compounds of the invention are also conjugated to soluble polymers that serve as targeted drug carriers. Such polymers include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspartamidephenol, or polyethylene oxide-polylysine substituted with palmitoyl residues. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers for controlled drug release, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polycaprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and crosslinked or amphipathic block copolymers of hydrogels.

Each unit dose of a dosage form (pharmaceutical composition) suitable for administration may contain from about 1mg to about 100mg of the active ingredient. In these pharmaceutical compositions, the weight of the active ingredient will generally be from about 0.5% to about 95% of the total weight of the composition.

Gelatin capsules may contain the active ingredient in combination with powder carriers such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Tablets and capsules can be manufactured as a sustained release product to provide a continuous release of drug over a period of time. The compressed tablets may be sugar coated or coated with a film to mask any unpleasant taste and to shield the tablet from the atmosphere, or enteric-coated for selective disintegration in the gastrointestinal tract.

Generally, water, a suitable oil, saline, hydrated dextrose (glucose), and related sugar solutions and glycols (e.g., propylene glycol or polyethylene glycol) are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain water-soluble salts of the active ingredient, suitable stabilizers, and possibly buffer substances. Antioxidants are suitable stabilizers, such as sodium bisulfite, sodium sulfite, or vitamin C, either alone or in combination, or citric acid and its salts and sodium EDTA salts. In addition, parenteral solutions also contain preservatives, such as benzalkonium chloride, methyl-or propyl-parabens, and chlorobutanol.

Wherein the compound of the invention is combined with other anticoagulant agents, e.g., a daily dose of about 0.1 to 100mg of the compound of formula (I) and about 1 to 7.5mg of the second anticoagulant per kilogram of patient body weight. For a tablet dosage form, the compound of the invention may generally be about 5 to 10mg per dosage unit and the amount of the second anti-agglomerating agent is from about 1 to 5mg per dosage unit. Other anti-coagulation agents specifically include, but are not limited to, apixaban, rivaroxaban, edoxaban, betrixaban, dabigatran, bemiparin, enoxaparin sodium, tinzaparin sodium, danaparin sodium, pentosan sodium, nadroparin calcium, aclidinarin sodium, heparan sodium, and the like.

In accordance with general guidelines, the compounds of the invention are administered in combination with an antiplatelet agent in a typical daily dose of from about 0.01 to 25mg of the compound of formula (I) and from about 50 to 150mg of the antiplatelet agent per kilogram of patient body weight, preferably from about 0.1 to 1mg of the compound of formula (I) and from about 1 to 3mg of the antiplatelet agent.

When the compound of formula (I) is administered in combination with a thrombolytic agent, a typical daily dose may be about 0.1 to 1mg of the compound of formula (I) per kg of patient body weight, and when the thrombolytic agent is administered together with the compound of formula (I) in the presence of the thrombolytic agent, the dose of the thrombolytic agent may be reduced by about 70-80% compared to the typical dose when the thrombolytic agent is administered alone.

When two or more of the foregoing second therapeutic agents are administered with a compound of formula (I), generally, the amount of each component in a typical daily dose and typical dosage form may be reduced relative to the usual dose when administered alone, taking into account the additive or synergistic effect of the therapeutic agents when administered in combination.

In particular, when provided as a single dosage unit, there is the possibility of a chemical reaction between the active ingredients of the combination. For this reason, when the compound of formula (I) or (II) and the second therapeutic agent are combined in a single dosage unit, they are formulated such that physical contact between the active ingredients is minimized (i.e., reduced), even though the active ingredients are combined in a single dosage unit. For example, one active ingredient may be an enteric coating. By enteric coating one active ingredient, it is possible not only to minimize contact between the combined active ingredients, but also to control the release of one of the ingredients in the gastrointestinal tract so that one of the components is not released in the stomach but in the small intestine. Further, the sustained release component may additionally be enteric coated to facilitate release of the component only in the intestinal tract. Yet another approach involves the formulation of a combination product in which one component is coated with a sustained and/or enteric release polymer and the other component is also coated with a polymer such as a low viscosity grade hydroxypropyl methylcellulose (HPMC) or other suitable material known in the art for the purpose of further isolating the active ingredients. The polymer coating forms an additional barrier to reaction with other components.

These and other methods of minimizing contact between the components of the combination product of the invention, whether they are administered in a single dosage form or in separate forms, but at the same time or in the same manner, will be apparent to those skilled in the art once apprised of the present disclosure.

The compound or the medicinal salt or the hydrate thereof can be effectively used for preventing, treating or relieving the thromboembolic diseases of patients, and particularly can be effectively used for treating myocardial infarction, angina, restenosis after reocclusion and angioplasty or aortic coronary artery bypass surgery, stroke, transient ischemic attack, peripheral arterial occlusive diseases, pulmonary embolism or deep vein thrombosis.

General Synthesis of Compounds of the invention

In general, the compounds of the invention may be prepared by the methods described herein, wherein the substituents are as defined in formula (I) or formula (II), unless otherwise indicated. The following reaction schemes and examples serve to further illustrate the context of the invention.

Those skilled in the art will recognize that: the chemical reactions described herein may be used to suitably prepare a number of other compounds of the invention, and other methods for preparing the compounds of the invention are considered to be within the scope of the invention. For example, the synthesis of those non-exemplified compounds according to the present invention can be successfully accomplished by those skilled in the art by modification, such as appropriate protection of interfering groups, by the use of other known reagents in addition to those described herein, or by some routine modification of reaction conditions. In addition, the reactions disclosed herein or known reaction conditions are also recognized as being applicable to the preparation of other compounds of the present invention.

The examples described below, unless otherwise indicated, are all temperatures set forth in degrees Celsius. Reagents were purchased from commercial suppliers such as Lingkai medicine, Aldrich Chemical Company, Inc., Arco Chemical Company and Alfa Chemical Company, and were used without further purification unless otherwise indicated. General reagents were purchased from Shantou Wen Long chemical reagent factory, Guangdong Guanghua chemical reagent factory, Guangzhou chemical reagent factory, Tianjin HaoLiyu Chemicals Co., Ltd, Qingdao Tenglong chemical reagent Co., Ltd, and Qingdao Kaseiki chemical plant.

The following reactions are generally carried out under positive pressure of nitrogen or argon or by sleeving a dry tube over an anhydrous solvent (unless otherwise indicated), the reaction vial being stoppered with a suitable rubber stopper and the substrate being injected by syringe. The glassware was dried.

The column chromatography is performed using a silica gel column. Silica gel (300 and 400 meshes) was purchased from Qingdao oceanic chemical plants. Nuclear magnetic resonance spectroscopy with CDC1₃Or DMSO-d₆As solvent (reported in ppm) TMS (0ppm) or chloroform (7.25ppm) was used as reference standard. When multiple peaks occur, the following abbreviations will be used: s (singleton), d (doublet), t (triplet ), m (multiplet, multiplet), br (broad ), dd (doublet of doublets, quartet), dt (doublet of triplets, doublet). Coupling constants are expressed in hertz (Hz).

The GC-MS is measured by an Agilent 7890A/5975C GC/MS mass spectrometer;

low resolution Mass Spectral (MS) data were measured by an Agilent6320 series LC-MS spectrometer equipped with a G1312A binary pump and a G1316A TCC (column temperature maintained at 30 ℃), a G1329A autosampler and a G1315B DAD detector were applied for analysis, and an ESI source was applied to the LC-MS spectrometer.

Low resolution Mass Spectral (MS) data were determined by Agilent6120 series LC-MS spectrometer equipped with a G1311A quaternary pump and a G1316A TCC (column temperature maintained at 30 ℃), a G1329A autosampler and a G1315D DAD detector were used for analysis, and an ESI source was used for the LC-MS spectrometer.

Both spectrometers were equipped with an Agilent Zorbax SB-C18 column, 2.1X 30mm, 5 μm. The injection volume is determined by the sample concentration; the flow rate is 0.6 mL/min; peaks of HPLC were recorded by UV-Vis wavelength at 210nm and 254 nm. The mobile phases were 0.1% formic acid in acetonitrile (phase a) and 0.1% formic acid in ultrapure water (phase B). Gradient elution conditions are shown in table 1:

TABLE 1

Time (min)	A(CH₃CN，0.1％HCOOH)	B(H₂O，0.1％HCOOH)
			0-3	5-100	95-0
3-6	100	0
			6-6.1	100-5	0-95
6.1-8	5	95

Compound purification was assessed by Agilent 1100 series High Performance Liquid Chromatography (HPLC) with UV detection at 210nm and 254nm, a Zorbax SB-C18 column, 2.1X 30mm, 4 μm, 10min, flow rate 0.6mL/min, 5-95% (0.1% formic acid in acetonitrile) in (0.1% formic acid in water), the column temperature was maintained at 40 ℃.

The following acronyms are used throughout the invention:

HATU 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate

Cbz benzyloxycarbonyl

DPPA Azidophosphoric acid Diphenyl ester

Ms methanesulfonyl

V/V, V/V volume ratio

M, mol/L mol/liter

Microliter of μ L

mL of

mmol millimole

g

mg of

The following synthetic schemes describe the steps for preparing the compounds disclosed herein.

Synthesis scheme 1

Compounds (I) can be prepared by the methods described in FIG. 1, wherein R is¹、R²、R³X, Y, Z, n have the meaning as indicated in the present invention. Carrying out nucleophilic substitution reaction on the compound (1) and the compound (2) under the action of alkali to obtain a compound (3); the compound (3) and the compound (4) are added in the presence of cuprous iodide and a ligand (such as N, N' -dimethylEthylenediamine) and a base (such as potassium carbonate) in a solvent at a high temperature to obtain a compound (5); deprotecting the compound (5) in ethanol under the action of methylamine to generate a compound (6); and carrying out condensation reaction on the compound (6) and the compound (7) to obtain the target compound (I).

Synthesis scheme 2

Compounds (I) can be prepared by the methods described in FIG. 2, wherein R is¹、R²、R³X, Y, Z, n have the meaning as indicated in the present invention. Reacting the compound (1) with the compound (8) under alkaline conditions to obtain a compound (9); hydrolyzing the compound (9) under alkaline conditions to obtain a compound (10); carrying out a curtis rearrangement reaction on the compound (10), DPPA and benzyl alcohol to obtain a compound (11); reacting the compound (11) with the compound (12) under the action of lithium bistrimethylsilyl amine to generate a compound (13); reacting the compound (13) with methylsulfonyl chloride under the action of alkali to obtain a compound (14); reacting the compound (14) with sodium azide to obtain a compound (15); under the action of triphenylphosphine, the compound (15) is subjected to reduction reaction in a proper solvent to generate a compound (16); and carrying out condensation reaction on the compound (16) and the compound (7) to obtain the target compound (I).

The compounds, pharmaceutical compositions and uses thereof provided by the present invention are further illustrated below in connection with the examples.

Examples

Example 1: (S) -5-chloro-N- ((2-oxo-3- (4- ((6-oxopyrimidin-1 (6H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1:3- ((5-bromothien-3-yl) methyl) pyrimidin-4 (3H) -one

To a solution of pyrimidin-4 (3H) -one (1.92g,20.0mmol) in methanol (100mL) was added potassium carbonate (5.52g,39.9mmol) and 2-bromo-4- (bromomethyl) thiophene (8.56g,20.1 mmol). The mixture was heated to 90 ℃ and stirred for 3 hours, filtered, and the solvent was evaporated under reduced pressure. Water (30mL) was added, and the mixture was extracted with ethyl acetate (40 mL. times.3). The organic phases were combined and dried over anhydrous sodium sulfate. Filtration and evaporation of the solvent under reduced pressure followed by column chromatography of the crude product (petroleum ether/ethyl acetate (v/v) ═ 2/1) gave a yellow solid (1.29g, 23.8%).

MS(ESI,pos.ion)m/z:272.90(M+1)。

Step 2 (S) -2- ((2-oxo-3- (4- ((6-oxopyrimidin-1 (6H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione

To the sealed tube were added 3- ((5-bromothien-3-yl) methyl) pyrimidin-4 (3H) -one (1.29g,4.76mmol), (R) -2- ((2-oxooxazolidin-5-yl) isoindoline-1, 3-dione (1.17g,4.75mmol), N' -dimethylethylenediamine (204 μ L,1.86mmol), cuprous iodide (180mg,0.95mmol), potassium carbonate (1.97g,14.3mmol), and 1, 4-dioxane (15mL) in this order, and under a nitrogen atmosphere, the mixture was heated to 120 ℃ and stirred for 10 hours, filtered, the solvent was evaporated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/5) to give a white solid (806mg, 38.8%).

MS(ESI,pos.ion)m/z:437.1(M+1)。

Step 3 (S) -5- (aminomethyl) -3- (4- ((6-oxopyrimidin-1 (6H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (S) -2- ((2-oxo-3- (4- ((6-oxopyrimidin-1 (6H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione (805mg,1.84mmol) in ethanol (19mL) was added 40% aqueous methylamine solution (1.9 mL). The mixture was heated to 95 ℃ and stirred for 1.5 h, cooled to room temperature, and the solvent was evaporated under reduced pressure, and the crude product was used in the next reaction without further treatment.

MS(ESI,pos.ion)m/z:307.0(M+1)。

Step 4 (S) -5-chloro-N- ((2-oxo-3- (4- ((6-oxopyrimidin-1 (6H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a solution of 5-chlorothiophene-2-carboxylic acid (359mg,2.21mmol) and N, N-diisopropylethylamine (1.2mL,7.3mmol) in dichloromethane (20mL) were added HATU (1.05g,2.76mmol) and (S) -5- (aminomethyl) -3- (4- ((6-oxopyrimidin-1 (6H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (565mg,1.84 mmol). The mixture was stirred at room temperature for 6 hours. The solvent was evaporated under reduced pressure, methylene chloride (50mL) was added to the residue, and the organic phase was washed with a saturated aqueous sodium hydrogencarbonate solution (25mL) and saturated brine (25mL) in this order and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) gave a white solid (100mg, 10%).

MS(ESI,pos.ion)m/z:451.0(M+1)；

¹H NMR(600MHz,DMSO-d₆):8.95(t,J＝5.8Hz,1H),8.60(s,1H),7.92(d,J＝6.6Hz,1H),7.68(d,J＝4.1Hz,1H),7.19(d,J＝4.0Hz,1H),7.03–6.98(m,1H),6.55(d,J＝1.6Hz,1H),6.43(dd,J＝6.6,0.6Hz,1H),5.04–4.95(m,2H),4.93–4.86(m,J＝11.8,5.6Hz,1H),4.13(t,J＝8.9Hz,1H),3.79(dd,J＝9.0,6.4Hz,1H),3.65–3.56(m,J＝14.4,8.7Hz,2H)。

Example 2: (S) -5-chloro-N- ((2-oxo-3- (4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1- ((5-bromothien-3-yl) methyl) pyridin-2 (1H) -one

To a solution of pyridin-2 (1H) -one (571mg,6mmol) in methanol (20mL) was added potassium carbonate (1.66g,12mmol) and 2-bromo-4- (bromomethyl) thiophene (2.3g,9 mmol). The mixture was heated to 70 ℃ and stirred for 3 hours, filtered and the solvent was evaporated off from the filtrate under reduced pressure. Water (30mL) was added and extracted with ethyl acetate (40 mL. times.3). The organic phases were combined, washed with saturated brine (20 mL. times.2), and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) gave a white solid (1.16g, 71.6%). MS (ESI, pos. ion) M/z 272.05(M + 1).

Step 2 (S) -2- ((2-oxo-3- (4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione

To the sealed tube were added 1- ((5-bromothien-3-yl) methyl) pyridin-2 (1H) -one (1.1g,4.0mmol), (R) -2- ((2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (1.0g,4.0mmol), N' -dimethylethylenediamine (144mg,1.6mmol), cuprous iodide (155mg,0.8mmol), potassium carbonate (1.89g,12.0mmol), and 1, 4-dioxane (20mL) in that order. The mixture was heated to 120 ℃ under nitrogen atmosphere and stirred for 10 hours. Cool to room temperature, filter, evaporate the solvent from the filtrate under reduced pressure, and purify the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/5) to give a white solid (950mg, 53.6%).

MS(ESI,pos.ion)m/z:436.10(M+1)。

Step 3 (S) -5- (aminomethyl) -3- (4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a suspension of (S) -2- ((2-oxo-3- (4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione (435mg,1.0mmol) in ethanol (10mL) was added 40% aqueous methylamine solution (1.0 mL). The mixture was heated to 95 ℃ and stirred for 1.5 hours, and the solvent was evaporated under reduced pressure. The crude product was used in the next step without purification.

Step 4 (S) -5-chloro-N- ((2-oxo-3- (4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a solution of 5-chlorothiophene-2-carboxylic acid (195mg,1.2mmol) in dichloromethane (20mL) was added N, N-diisopropylethylamine (380 μ L,2.0mmol), HATU (570mg,1.5mmol) and (S) -5- (aminomethyl) -3- (4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (305mg,1.0mmol) in that order. The mixture was stirred at room temperature for 6 hours. The solvent was evaporated under reduced pressure, methylene chloride (15mL) was added to the residue, and the organic phase was washed with saturated sodium hydrogencarbonate (10mL) and saturated brine (10mL) in this order and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/4) gave a white solid (250mg, 55.6%).

MS(ESI,pos.ion)m/z:450.00(M+1)；

¹H NMR(600MHz,DMSO-d₆):8.94(t,J＝5.8Hz,1H),7.73(dd,J＝6.8,1.7Hz,1H),7.67(d,J＝4.1Hz,1H),7.40(ddd,J＝8.9,6.6,2.1Hz,1H),7.18(d,J＝4.0Hz,1H),6.97–6.92(m,1H),6.51(d,J＝1.6Hz,1H),6.40(d,J＝8.7Hz,1H),6.22(td,J＝6.7,1.3Hz,1H),4.99(d,J＝14.2Hz,1H),4.95(d,J＝14.2Hz,1H),4.88(td,J＝11.7,5.6Hz,1H),4.11(t,J＝8.9Hz,1H),3.78(dd,J＝8.9,6.4Hz,1H),3.60(t,J＝5.6Hz,2H)。

Example 3: (S) -5-chloro-N- ((2-oxo-3- (4- ((2-oxopyrimidin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1- ((5-bromothien-3-yl) methyl) pyrimidin-2 (1H) -one

To a solution of pyrimidin-2 (1H) -one hydrogen chloride (1.92g,20.0mmol) in methanol (100mL) was added potassium carbonate (8.28g,59.9mmol) and 2-bromo-4- (bromomethyl) thiophene (8.52g,20.0 mmol). The mixture was heated to 90 ℃ and stirred for 3 hours, filtered, and the solvent was evaporated under reduced pressure. Water (30mL) was added, and the mixture was extracted with ethyl acetate (40 mL. times.3). The organic phases were combined and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (acetone/dichloromethane (v/v) ═ 1/20) gave a white solid (1.34g, 24.7%).

MS(ESI,pos.ion)m/z:272.90(M+1)。

Step 2 (S) -2- ((2-oxo-3- (4- ((2-oxopyrimidin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione

To a sealed tube were added in order 1- ((5-bromothien-3-yl) methyl) pyrimidin-2 (1H) -one (1.34g,4.94mmol), (R) -2- ((2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (1.22g,4.95mmol), N' -dimethylethylenediamine (217 μ L,2.0mmol), cuprous iodide (190mg,1.0mmol), potassium carbonate (2.05g,14.8mmol), and 1, 4-dioxane (15mL), and the mixture was heated to 120 ℃ under a nitrogen atmosphere and stirred for 10 hours. Cooled to room temperature, water (20mL) was added, and the mixture was extracted with ethyl acetate (40 mL. times.3). The organic phases were combined, washed with saturated brine (30 mL. times.3), and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (acetone/dichloromethane (v/v) ═ 1/7) gave a white solid (640mg, 30%).

MS(ESI,pos.ion)m/z:437.0(M+1)。

Step 3 (S) -5- (aminomethyl) -3- (4- ((2-oxopyrimidin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (S) -2- ((2-oxo-3- (4- ((2-oxopyrimidin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione (640mg,1.5mmol) in ethanol (15mL) was added 40% aqueous methylamine solution (1.5 mL). The mixture was heated to 95 ℃ and stirred for 1.5 h, cooled to room temperature, and the solvent was evaporated under reduced pressure, and the crude product was used in the next reaction without further treatment.

Step 4 (S) -5-chloro-N- ((2-oxo-3- (4- ((2-oxopyrimidin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a solution of 5-chlorothiophene-2-carboxylic acid (290mg,1.8mmol) and N, N-diisopropylethylamine (900 μ L,5mmol) in dichloromethane (20mL) was added HATU (840mg,2.2mmol) and (S) -5- (aminomethyl) -3- (4- ((2-oxopyrimidin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (450mg,1.5 mmol). The mixture was stirred at room temperature for 6 hours. The solvent was distilled off under reduced pressure, methylene chloride (50mL) was added thereto, and the organic phase was washed successively with a saturated aqueous sodium hydrogencarbonate solution (25mL) and saturated brine (25mL), and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) gave a white solid (100mg, 20%).

MS(ESI,pos.ion)m/z:450.9(M+1)；

¹H NMR(600MHz,DMSO-d₆):8.95(t,J＝5.8Hz,1H),8.55(dd,J＝4.1,2.9Hz,1H),8.27(dd,J＝6.5,2.8Hz,1H),7.68(d,J＝4.1Hz,1H),7.19(d,J＝4.0Hz,1H),7.03–7.00(m,1H),6.55(d,J＝1.6Hz,1H),6.45(dd,J＝6.5,4.1Hz,1H),4.98–4.92(m,2H),4.92–4.87(m,J＝11.7,7.6,4.5Hz,1H),4.12(t,J＝8.9Hz,1H),3.79(dd,J＝9.0,6.4Hz,1H),3.61(t,J＝5.6Hz,2H)。

Example 4: (S) -5-chloro-N- ((2-oxo-3- (4- ((2-oxopyrazin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1- ((5-bromothien-3-yl) methyl) pyrazin-2 (1H) -one

To a solution of pyrazin-2 (1H) -one (2.50g,26.0mmol) in methanol (20mL) was added potassium carbonate (7.17g,51.9mmol) and 2-bromo-4- (bromomethyl) thiophene (11.13g,26.09 mmol). The mixture was heated to 85 ℃ and stirred for 3 hours, filtered, and the solvent was evaporated from the filtrate under reduced pressure. Water (30mL) was added, followed by ethyl acetate (30 mL. times.3). The organic phases were combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2/1) gave a yellow solid (850mg, 12%).

MS(ESI,pos.ion)m/z:272.90(M+1)。

Step 2 (S) -2- ((2-oxo-3- (4- ((2-oxopyrazin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione

To the sealed tube were added in this order 1- ((5-bromothien-3-yl) methyl) pyrazin-2 (1H) -one (850mg,3.14mmol), (R) -2- ((2-oxooxazolidin-5-yl) isoindoline-1, 3-dione (771mg,3.13mmol), N' -dimethylethylenediamine (110mg,1.25mmol), cuprous iodide (119mg,0.625mmol), potassium carbonate (1.3g,9.4mmol), and 1, 4-dioxane (10mL), and the mixture was heated to 120 ℃ under a nitrogen atmosphere, stirred for 10 hours, filtered, the solvent was evaporated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) to give a white solid (260mg, 19%).

MS(ESI,pos.ion)m/z:436.9(M+1)。

Step 3 (S) -5- (aminomethyl) -3- (4- ((2-oxopyrazin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (S) -2- ((2-oxo-3- (4- ((2-oxopyrazin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) isoindoline-1, 3-dione (260mg,0.60mmol) in ethanol (6mL) was added 40% aqueous methylamine (0.6 mL). The mixture was heated to 95 ℃ and stirred for 1.5 hours, cooled to room temperature, and the solvent was evaporated under reduced pressure, and the crude product was used in the next reaction without further treatment.

MS(ESI,pos.ion)m/z:307.10(M+1)。

Step 4 (S) -5-chloro-N- ((2-oxo-3- (4- ((2-oxopyrazin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a solution of 5-chlorothiophene-2-carboxylic acid (120mg,0.74mmol) and N, N-diisopropylethylamine (460 μ L,2.8mmol) in dichloromethane (20mL) was added HATU (340mg,0.89mmol) and (S) -5- (aminomethyl) -3- (4- ((2-oxopyrazin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (182mg,0.594 mmol). The mixture was stirred at room temperature for 6 hours. The solvent was evaporated under reduced pressure, and methylene chloride (50mL) was added to the residue, which was washed with a saturated aqueous sodium hydrogencarbonate solution (30mL) and brine (30mL) in this order, and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) gave a white solid (140mg, 52%).

MS(ESI,pos.ion)m/z:450.7(M+1)；

¹H NMR(600MHz,DMSO-d₆):8.95(t,J＝5.8Hz,1H),8.04(d,J＝1.0Hz,1H),7.71(dd,J＝4.4,1.0Hz,1H),7.67(d,J＝4.1Hz,1H),7.34(d,J＝4.4Hz,1H),7.19(d,J＝4.1Hz,1H),7.04–7.03(m,1H),6.53(d,J＝1.6Hz,1H),5.01–4.93(m,2H),4.93–4.86(m,J＝11.8,5.6Hz,1H),4.12(t,J＝8.9Hz,1H),3.79(dd,J＝9.0,6.4Hz,1H),3.65–3.56(m,2H)。

Example 5: (S) -5-chloro-N- ((3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1 3-chloro-4-methylthiophene-2-carboxylic acid methyl ester

To a 6M aqueous solution (50mL) of 3-amino-4-methylthiophene-2-carboxylic acid methyl ester (20g,12mmol) in hydrochloric acid at 0 ℃ was added a solution of sodium nitrite (8g,120mmol) in water (20mL), and after the addition was completed, the mixture was stirred for 1 hour, and then a solution of cuprous chloride (12g,120mmol) in 12M hydrochloric acid (50mL) was slowly added. After completion of the dropwise addition, the mixture was warmed to room temperature, water (100mL) was added, and the mixture was extracted with ethyl acetate (100 mL. times.2). The organic phases were combined, washed successively with water (50 mL. times.2) and saturated brine (50 mL. times.2), and dried over anhydrous sodium sulfate. Filtering, and evaporating the solvent under reduced pressure to obtain yellow solid which is directly used in the next step without purification.

GC-MS:M＝189.98。

Step 2-4- (bromomethyl) -3-chlorothiophene-2-carboxylic acid methyl ester

To a round bottom flask were added 3-chloro-4-methylthiophene-2-carboxylic acid methyl ester (35g,185mmol), N-bromosuccinimide (33.8g,203mmol), benzoyl peroxide (4.84g,20mmol), and carbon tetrachloride (200mL) in that order. The mixture was heated to 85 ℃ and stirred for 7 hours. Filtration was carried out, the solvent was evaporated from the filtrate under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 20/1) to give a yellow oil (22g, 75%).

GC-MS:M＝267.90。

Step 3 methyl 3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylate

To a solution of pyridin-2 (1H) -one (8.09g,30mmol) and methyl 4- (bromomethyl) -3-chlorothiophene-2-carboxylate (2.38g,25.0mmol) in acetone (100mL) was added potassium carbonate (63.6g,460.2 mmol). The mixture was heated to 60 ℃ and stirred overnight, cooled to room temperature. Suction filtration was carried out, the solvent was evaporated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 4/1) to give a white solid (6.38g, 89.9%).

MS(ESI,pos.ion)m/z:284.0(M+1)。

Step 4 3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylic acid

To a solution of methyl 3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylate (5.93g,20.9mmol) in methanol (120mL) was added a solution of lithium hydroxide monohydrate (2.63g,62.7mmol) in water (40mL), and the mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure, the residue was dissolved in water (100mL), extracted with ethyl acetate (30mL × 3), and the aqueous phase was acidified with concentrated hydrochloric acid to pH 2. The solid precipitated, filtered off with suction, washed with water and dried to give a white solid (4.9g, 87.0%).

MS(ESI,pos.ion)m/z:270.0(M+1)。

Step 5 benzyl (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) carbamate

To a solution of 3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylic acid (1.35g,5.0mmol), benzyl alcohol (624. mu.L, 6.0mmol), and triethylamine (1.39mL,10mmol) in 1, 4-dioxane (15mL) was added diphenyl phosphorazide (1.43mL,7.5mmol), and the mixture was heated to 80 ℃ and stirred for 5.5 hours. The reaction solution was cooled to room temperature, water (30mL) was added, extraction was performed with ethyl acetate (40 mL. times.3), and the organic phases were combined, washed with water (30 mL. times.3) and saturated brine (50mL) in this order, and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/1) gave a yellow solid (1.17g, 62.6%).

MS(ESI,pos.ion)m/z:374.90(M+1)。

Step 6 (R) -3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -5- (hydroxymethyl) oxazolidin-2-one

To a solution of benzyl (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) carbamate (750mg,2.001mmol) in anhydrous tetrahydrofuran (10mL) was slowly added dropwise a solution of 1M lithium bistrimethylsilyl amide in tetrahydrofuran (3.002mL,3.002mmol) under nitrogen at-78 ℃. The mixture was stirred at-78 ℃ for 0.5 hour, then warmed to 0 ℃ and stirred for 15 minutes, and then a solution of (R) -oxiran-2-ylmethylbutyrate (432.7mg,3.002mmol) in tetrahydrofuran (4mL) was slowly added dropwise. The mixture was cooled to-78 ℃ and stirred for 2 hours, then warmed to room temperature and stirred overnight. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a white solid (370mg, 54%).

MS(ESI,pos.ion)m/z:340.90(M+1)。

Step 7 (R) - (3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl methanesulfonate

To a round bottom flask was added (R) -3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -5- (hydroxymethyl) oxazolidin-2-one (374mg,1.10mmol), N-diisopropylethylamine (310 μ L,2.19mmol), 4-dimethylaminopyridine (13.4mg,0.110mmol), and dichloromethane (10 mL). The mixture was cooled to 0 ℃ and methanesulfonyl chloride (127. mu.L, 1.65mmol) was slowly added dropwise. The reaction solution was stirred at room temperature for 4 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a white solid (407mg, 88.5%).

MS(ESI,pos.ion)m/z:418.85(M+1)。

Step 8 (R) -5- (azidomethyl) -3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (R) - (3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl methanesulfonate (400mg,0.9549mmol) in N, N-dimethylformamide (20mL) was added sodium azide (931.2mg,14.32mmol), and the mixture was heated to 70 ℃ and stirred for 12 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (dichloromethane/methanol (v/v) ═ 20/1) to give a white solid (340mg, 97%).

MS(ESI,pos.ion)m/z:365.90(M+1)。

Step 9 (S) -5- (aminomethyl) -3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (R) -5- (azidomethyl) -3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (340mg,0.93mmol) in tetrahydrofuran (20mL) was added water (2mL) and triphenylphosphine (490mg,1.9mmol), the reaction solution was stirred overnight at room temperature, and the solvent was evaporated under reduced pressure. The crude product was purified by column chromatography (dichloromethane/methanol (v/v) ═ 10/1) to give a colourless oil (330mg, 100%).

MS(ESI,pos.ion)m/z:339.95(M+1)。

Step 10 (S) -5-chloro-N- ((3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a round bottom flask was added (S) -5- (aminomethyl) -3- (3-chloro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (340mg,1.001mmol), 5-chlorothiophene-2-carboxylic acid (211.5mg,1.301mmol), N-diisopropylethylamine (331 μ L,2.001mmol), HATU (627.6mg,1.601mmol) and dichloromethane (30 mL). The reaction was stirred at room temperature overnight, the solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a white solid (220mg, 45%).

MS(ESI,pos.ion)m/z:483.70(M+1)；

¹H NMR(600MHz,CDCl₃)7.41(d,J＝4.0Hz,1H),7.38(t,J＝7.1Hz,2H),7.27(s,1H),7.23(s,1H),6.88(d,J＝3.9Hz,1H),6.63(d,J＝9.4Hz,1H),6.22(t,J＝6.5Hz,1H),5.03(s,2H),4.87-4.96(m,1H),4.11(t,J＝8.9Hz,1H),3.89(dd,J＝8.9,6.2Hz,1H),3.80-3.83(m,2H)。

Example 6 (S) -5-chloro-N- ((3- (4- ((3-fluoro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1- ((5-bromothien-3-yl) methyl) -3-fluoropyridin-2 (1H) -one

To a solution of 3-fluoropyridin-2 (1H) -one (2.26g,20.0mmol) in methanol (100mL) was added potassium carbonate (5.52g,39.9mmol) and 2-bromo-4- (bromomethyl) thiophene (8.52g,20.0 mmol). The mixture was heated to 85 ℃ and stirred for 3 hours, filtered, and the solvent was evaporated from the filtrate under reduced pressure. Water (30mL) was added, and the mixture was extracted with ethyl acetate (40 mL. times.3). The organic phases were combined, washed with saturated brine (50mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 4/1) gave a white solid (2.0g, 35%).

MS(ESI,pos.ion)m/z:289.80(M+1)。

Step 2 (S) -2- ((3- (4- ((3-fluoro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione

To the sealed tube were added in this order 1- ((5-bromothien-3-yl) methyl) -3-fluoropyridin-2 (1H) -one (2.0g,6.9mmol), (R) -2- ((2-oxooxazolidin-5-yl) isoindoline-1, 3-dione (1.71g,6.95mmol), N' -dimethylethylenediamine (244mg,2.77mmol), cuprous iodide (270mg,1.4mmol), potassium carbonate (2.88g,20.8mmol), and 1, 4-dioxane (20mL), and the mixture was heated to 120 ℃ under a nitrogen atmosphere and stirred for 10 hours, filtered, the solvent was evaporated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2/3) to give a white solid (860mg, 27%).

MS(ESI,pos.ion)m/z:453.90(M+1)。

Step 3 (S) -5- (aminomethyl) -3- (4- ((3-fluoro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (S) -2- ((3- (4- ((3-fluoro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (860mg,1.9mmol) in ethanol (20mL) was added 40% aqueous methylamine solution (2.0 mL). The mixture was heated to 95 ℃ and stirred for 1.5 h, cooled to room temperature, and the solvent was evaporated under reduced pressure, and the crude product was used in the next reaction without further treatment.

Step 4 (S) -5-chloro-N- ((3- (4- ((3-fluoro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a solution of (S) -5- (aminomethyl) -3- (4- ((3-fluoro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (610mg,1.9mmol) and N, N-diisopropylethylamine (978mg,7.56mmol) in dichloromethane (20mL) was added HATU (1.08g,2.84mmol) and 5-chlorothiophene-2-carboxylic acid (368mg,2.26 mmol). The mixture was stirred at room temperature for 6 hours. The solvent was evaporated under reduced pressure, methylene chloride (50mL) was added thereto, and the mixture was washed with a saturated aqueous sodium hydrogencarbonate solution (30mL) and brine (30mL) in this order and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) gave a white solid (200mg, 22.7%).

MS(ESI,pos.ion)m/z:468.0(M+1)；

¹H NMR(600MHz,DMSO-d₆):8.95(t,J＝5.8Hz,1H),7.68(t,J＝5.4Hz,1H),7.63(d,J＝6.8Hz,1H),7.42–7.36(m,1H),7.19(d,J＝4.0Hz,1H),6.99(d,J＝1.3Hz,1H),6.52(d,J＝1.6Hz,1H),6.22(td,J＝7.2,4.7Hz,1H),5.10–5.01(m,2H),4.93–4.86(m,J＝11.6,5.6Hz,1H),4.12(t,J＝8.9Hz,1H),3.79(dd,J＝8.9,6.4Hz,1H),3.61(t,J＝5.6Hz,2H)。

Example 7 (S) -5-chloro-N- ((3- (4- ((3-hydroxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1- ((5-bromothien-3-yl) methyl) -3-methoxypyridin-2 (1H) -one

To a solution of 3-methoxypyridin-2 (1H) -one (630mg,5.0mmol) in acetone (10mL) was added potassium carbonate (1.4g,10mmol) and 2-bromo-4- (bromomethoxy) thiophene (1.3g,5.0 mmol). The mixture was heated to 65 ℃ and stirred for 2.5 hours. Filtering, and evaporating the solvent from the filtrate under reduced pressure. Water (15mL) was added thereto, and the mixture was extracted with ethyl acetate (10 mL. times.4). The organic phases were combined, washed with saturated brine (20mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/1) gave a yellow oil (950mg, 63%).

MS(ESI,pos.ion)m/z:302.00(M+1)。

Step 2 (S) -2- ((3- (4- ((3-methoxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione

To a sealed tube were added in this order 1- ((5-bromothien-3-yl) methyl) -3-methoxypyridin-2 (1H) -one (4.4g,15mmol), (R) -2- ((2-oxooxazolidin-5-yl) isoindoline-1, 3-dione (3.6g,15mmol), N' -dimethylethylenediamine (520mg,5.9mmol), cuprous iodide (560mg,2.9mmol), potassium carbonate (6.1g,44mmol) and 1, 4-dioxane (20mL), and under a nitrogen atmosphere, the mixture was heated to 120 ℃ and stirred for 8 hours, cooled to room temperature, filtered, the solvent was evaporated under reduced pressure from the filtrate, and the crude product was purified by column chromatography (dichloromethane/methanol (v/v) ═ 50/1) to give a white solid (2.51g, 37%).

MS(ESI,pos.ion)m/z:466.00(M+1)。

Step 3 (S) -2- ((3- (4- ((3-hydroxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione

Boron tribromide (110 μ L,1.2mmol) was slowly added to a solution of (S) -2- ((3- (4- ((3-methoxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (470mg,1.0mmol) in dry dichloromethane (20mL) at-30 ℃ under a nitrogen atmosphere. The reaction solution was warmed to room temperature and stirred for 48 hours. Cooled to-30 ℃ and methanol (5mL) was slowly added dropwise thereto. The solvent was evaporated under reduced pressure, adjusted to pH 7 with 1M sodium hydroxide solution, and extracted with chloroform (30mL × 3). The organic phases were combined, washed with saturated brine (45mL) and dried over anhydrous sodium sulfate. Filtering, and evaporating the solvent under reduced pressure. The crude product was purified by column chromatography (dichloromethane/methanol (v/v) ═ 20/1) to give a yellow solid (300mg, 70%).

MS(ESI,pos.ion)m/z:452.10(M+1)。

Step 4 (S) -5- (aminomethyl) -3- (4- ((3-hydroxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a suspension of (S) -2- ((3- (4- ((3-hydroxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (1.08g,2.39mmol) in ethanol (30mL) was added 40% aqueous methylamine solution (3.0 mL). The mixture was heated to 95 ℃ and stirred for 1.5 hours, and the solvent was evaporated under reduced pressure. The crude product was used in the next step without purification.

Step 5 (S) -5-chloro-N- ((3- (4- ((3-hydroxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a solution of 5-chlorothiophene-2-carboxylic acid (360mg,2.2mmol) and (S) -5- (aminomethyl) -3- (4- ((3-hydroxy-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (710mg,2.2mmol) in dichloromethane (40mL) was added N, N-diisopropylethylamine (460 μ L,2.6mmol) and HATU (1.0g,2.6mmol) in that order. The mixture was stirred at room temperature for 6 hours. The solvent was evaporated under reduced pressure, and methylene chloride (50mL) was added to the residue, which was washed with saturated sodium hydrogencarbonate (25mL) and saturated brine (25mL) in this order, and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and column chromatography of the crude product (dichloromethane/methanol (v/v) ═ 40/1) gave a pale yellow solid (360mg, 35%).

MS(ESI,pos.ion)m/z:466.00(M+1)；

¹H NMR(600MHz,DMSO-d₆):8.94(t,J＝5.5Hz,1H),7.67(d,J＝3.9Hz,1H),7.20(d,J＝6.8Hz,1H),

7.18(d,J＝3.9Hz,1H),6.95(s,1H),6.68(d,J＝7.1Hz,1H),6.51(s,1H),6.10(t,J＝7.0Hz,1H),5.03(d,J＝14.3Hz,1H),5.00(d,J＝14.3Hz,1H),4.93–4.84(m,1H),4.10(t,J＝8.8Hz,1H),3.80–3.75(m,1H),3.60(t,J＝5.6Hz,2H)。

Example 8 (S) -1- ((5- (5- ((5-chlorothiophene-2-carboxamido) methyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid

Step 1: 2-oxo-1, 2-dihydropyridine-3-methyl-tert-butyl ester

To a solution of 2-oxo-1, 2-dihydropyridine-3-carboxylic acid (5.00g,35.9mmol) in t-butanol (100mL) were added di-tert-butyl dicarbonate (9.23g,42.3mmol) and N, N-dimethylpyridin-4-amine (1.34g,11.0mmol), and the mixture was heated to 40 ℃ and stirred overnight. Water (50mL) was added, and the mixture was extracted with ethyl acetate (50 mL. times.3). The organic phases were combined, washed with saturated brine (60mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and column chromatography of the crude product (dichloromethane/methanol (v/v) ═ 20/1) gave a white solid (1.70g, 24.2%).

¹H NMR(600MHz,DMSO-d₆):11.96(s,1H),7.92(dd,J＝7.0,2.1Hz,1H),7.60(dd,J＝6.2,2.0Hz,1H),6.25(dt,J＝13.3,9.0Hz,1H),1.48(s,9H)。

Step 2- ((5-bromothien-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid tert-butyl ester

To a round bottom flask was added 2-oxo-1, 2-dihydropyridine-3-methyl-tert-butyl ester (1.70g,8.71mmol), 2-bromo-4- (bromomethyl) thiophene (2.45g,9.57mmol), potassium carbonate (2.41g,17.4mmol) and acetone (30 mL). The mixture was heated to 65 ℃ and stirred for 4 hours, the solvent was evaporated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10/1) to give a yellow solid (1.83g, 56.8%).

MS(ESI,pos.ion)m/z:313.9(M-56)。

Step 3 (S) -1- ((5- (5- ((1, 3-dioxoisoindolin-2-yl) methyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid tert-butyl ester

To the sealed tube were added tert-butyl 1- ((5-bromothien-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylate (1.83g,4.94mmol), (R) -2- ((2-oxooxazolidin-5-yl) isoindoline-1, 3-dione (1.22g,4.95mmol), N' -dimethylethylenediamine (175mg,1.99mmol), cuprous iodide (188mg,0.982mmol), potassium carbonate (2.05g,14.8mmol), and 1, 4-dioxane (20mL) in this order, the mixture was heated to 120 ℃ under a nitrogen atmosphere, stirred for 10 hours, cooled to room temperature, filtered, the filtrate was evaporated under reduced pressure to remove the solvent, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2), a white solid (1.28g, 48.4%) was obtained.

¹H NMR(400MHz,DMSO-d₆):8.03(dd,J＝6.7,2.1Hz,1H),7.95–7.83(m,5H),6.97(s,1H),6.52(d,J＝1.5Hz,1H),6.30(t,J＝6.9Hz,1H),5.08–4.95(m,3H),4.16(t,J＝9.0Hz,1H),4.02(dd,J＝14.4,7.1Hz,1H),3.97–3.85(m,2H),1.49(s,9H)。

Step 4 (S) -1- ((5- (5- (aminomethyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid tert-butyl ester

To a suspension of (S) -1- ((5- (5- ((1, 3-dioxoisoindolin-2-yl) methyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid tert-butyl ester (1.28g,2.39mmol) in ethanol (30mL) was added 40% aqueous methylamine solution (2.4 mL). The mixture was heated to 95 ℃ and stirred for 1.5 hours, and the solvent was evaporated under reduced pressure. The crude product was used in the next step without purification.

Step 5 (S) -1- ((5- (5- ((5-chlorothiophene-2-carboxamido) methyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid tert-butyl ester

To a solution of 5-chlorothiophene-2-carboxylic acid (466mg,2.87mmol) and N, N-diisopropylethylamine (1.24g,9.58mmol) in dichloromethane (50mL) was added HATU (1.36g,3.58mmol) at room temperature. After stirring for 1 hour, (S) -1- ((5- (5- (aminomethyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid tert-butyl ester (969mg,2.39mmol) was added. The mixture was stirred at room temperature for 6 hours. The solvent was distilled off under reduced pressure, water (20mL) was added, and the mixture was extracted with methylene chloride (20 mL. times.3). The organic phases were combined, washed with saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/4) gave a white solid (1.21g, 92.1%).

¹H NMR(400MHz,CDCl3):8.11(d,J＝5.3Hz,1H),7.62(d,J＝5.3Hz,1H),7.41(d,J＝4.1Hz,1H),6.91(d,J＝4.0Hz,1H),6.78(s,1H),6.50(s,1H),6.31(t,J＝6.9Hz,2H),5.06(s,2H),5.01–4.92(m,J＝11.8,5.8Hz,1H),4.20–4.12(m,1H),3.89(dd,J＝9.4,6.4Hz,1H),3.83–3.70(m,2H),1.57(s,9H)。

Step 6 (S) -1- ((5- (5- ((5-chlorothiophene-2-carboxamido) methyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid

To a solution of (S) -1- ((5- (5- ((5-chlorothiophene-2-carboxamido) methyl) -2-oxooxazolidin-3-yl) thiophen-3-yl) methyl) -2-oxo-1, 2-dihydropyridine-3-carboxylic acid tert-butyl ester (1.21g,2.20mmol) in dichloromethane (50mL) was added trifluoroacetic acid (10mL,129 mmol). The mixture was stirred at room temperature overnight, the solvent was evaporated under reduced pressure, water (15mL) was added to precipitate a solid, which was filtered with suction and purified by column chromatography (dichloromethane/methanol (v/v) ═ 20/1) to give a red solid (36mg, 3.3%).

MS(ESI,neg.ion)m/z:491.9(M-1)；

¹H NMR(400MHz,DMSO-d₆):8.94(t,J＝5.6Hz,1H),8.40(dd,J＝7.3,2.0Hz,1H),8.34(dd,J＝6.6,2.0Hz,1H),7.67(d,J＝4.1Hz,1H),7.19(d,J＝4.1Hz,1H),7.06(s,1H),6.76(t,J＝6.9Hz,1H),6.56(d,J＝1.6Hz,1H),5.21(s,2H),4.97–4.85(m,J＝8.6,6.0Hz,1H),4.13(t,J＝9.0Hz,1H),3.79(dd,J＝8.9,6.5Hz,1H),3.65–3.56(m,J＝5.5Hz,2H).

Example 9: (S) -5-chloro-N- ((3- (3-nitrile-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1:1- (2-oxopropyl) pyridin-2 (1H) -one

To a round-bottom flask were added 1-chloroprop-2-one (11.10g,120mmol), pyridin-2 (1H) -one (9.51g,100mmol), potassium iodide (3.32g,20mmol), potassium carbonate (33.50g,240mmol) and acetone (150mL) in that order. The mixture was heated to 65 ℃ and stirred for 6 hours. Cool to room temperature, filter, evaporate the solvent from the filtrate under reduced pressure, and purify the crude product by column chromatography (ethyl acetate) to give a yellow oil (13.5g, 89.3%).

Step 2-2-amino-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-3-carbonitrile

To a solution of 1- (2-oxopropyl) pyridin-2 (1H) -one (13.50g,89.31mmol) and malononitrile (6.49g,98.24mmol) in ethanol (150mL) was added sulfur (3.437g,107.2mmol) and morpholine (16.34g,187.6mmol), and the mixture was heated to 85 ℃ and stirred for 2 hours. Cooled to room temperature and the solvent was evaporated under reduced pressure. Water (200mL) was added, and the mixture was extracted with ethyl acetate (100 mL. times.4). The organic phases were combined, washed with saturated brine (150mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/1) gave a yellow solid (2.3g, 11%).

MS(ESI,pos.ion)m/z:232.00(M+1)。

Step 3 benzyl (3-cyano-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) carbamate

To a round bottom flask was added 2-amino-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-3-carbonitrile (2.10g,9.079mmol) and pyridine (30mL), the mixture was stirred at room temperature for 15 minutes, then benzyl chloroformate (7.74mL,54.47mmol) was added slowly and stirred at room temperature overnight. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/1) to give a yellow solid (500mg, 20%).

MS(ESI,pos.ion)m/z:366.10(M+1)。

Step 4 (R) -2- (5- (hydroxymethyl) -2-oxooxazolidin-3-yl) -4- ((2-oxazolidin-1 (2H) -yl) methyl) thiophene-3-carbonitrile

To a solution of benzyl (3-cyano-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) carbamate (500mg,1.368mmol) in anhydrous tetrahydrofuran (10mL) under nitrogen protection at-78 ℃ was slowly added dropwise a solution of lithium bistrimethylsilyl amide in tetrahydrofuran (2.737mL,2.737mmol, 1M). The mixture was stirred at-78 ℃ for 0.5 hour, then warmed to 0 ℃ and stirred for 15 minutes, then cooled to-78 ℃ and then a solution of (R) -oxiran-2-ylmethyl butyrate (394.6mg,2.737mmol) in tetrahydrofuran (4mL) was slowly added dropwise. The mixture was stirred at-78 ℃ for 2 hours, warmed to room temperature and stirred overnight. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) to give a white solid (400mg, 88.2%).

MS(ESI,pos.ion)m/z:332.10(M+1)。

Step 5 (R) - (3- (3-nitrile-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl methanesulfonate

To a round bottom flask was added (R) -2- (5- (hydroxymethyl) -2-oxooxazolidin-3-yl) -4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-3-carbonitrile (91mg,0.2746mmol), N-diisopropylethylamine (272 μ L,1.648mmol), and dichloromethane (10 mL). The mixture was cooled to 0 ℃ and methanesulfonyl chloride (63.8. mu.L, 0.8238mmol) was slowly added dropwise. Stirred at room temperature for 4 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a white solid (445mg, 90.0%).

MS(ESI,pos.ion)m/z:410.05(M+1)。

Step 6 (R) -2- (5- (azidomethyl) -2-oxooxazolidin-3-yl) -4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-3-carbonitrile

To a solution of (R) - (3- (3-nitrile-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl methanesulfonate (105mg,0.2565mmol) in N, N-dimethylformamide (10mL) was added sodium azide (166.7mg,2.565mmol), and the mixture was heated to 70 ℃ and stirred for 12 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (dichloromethane/methanol (v/v) ═ 20/1) to give a colorless oil (65mg, 71.1%).

MS(ESI,pos.ion)m/z:357.10(M+1)。

Step 7 (S) -2- (5- (aminomethyl) -2-oxooxazolidin-3-yl) -4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-3-carbonitrile

To a solution of (R) -2- (5- (azidomethyl) -2-oxooxazolidin-3-yl) -4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-3-carbonitrile (65mg,0.1824mmol) in tetrahydrofuran (20mL) was added water (2mL) and triphenylphosphine (191.4mg,0.7295mmol), the mixture was stirred at room temperature overnight and the solvent was evaporated under reduced pressure. The crude product was purified by column chromatography (dichloromethane/methanol (v/v) ═ 10/1) to give a yellow oil (60mg, 99.6%). The product was used directly in the next step.

Step 8 (S) -5-chloro-N- (3- ((3-cyano-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) amino) -2-hydroxypropyl) thiophene-2-carboxamide

To a solution of (S) -2- (5- (aminomethyl) -2-oxooxazolidin-3-yl) -4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-3-carbonitrile (60mg,0.1816mmol) and N, N-diisopropylethylamine (120 μ L,0.7264mmol) in dichloromethane (20mL) at 0 ℃ was added a solution of 5-chlorothiophene-2-carbonyl chloride (270mg,1.5mmol) in dichloromethane (5 mL). The mixture was stirred at room temperature for 12 hours. Dichloromethane (50mL) was added and the mixture was washed with a 1M aqueous solution of sodium hydroxide (20 mL. times.2). The organic phase was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a colourless oil (65mg, 79.7%). The product was used directly in the next step.

Step 9 (S) -5-chloro-N- ((3- (3-nitrile-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a round bottom flask was added in sequence (S) -5-chloro-N- (3- ((3-cyano-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) amino) -2-hydroxypropyl) thiophene-2-carboxamide (65mg,0.145mmol), N' -carbonyldiimidazole (93.9mg,0.579mmol), 4-dimethylaminopyridine (3.6mg,0.0290mmol), and tetrahydrofuran (10 mL). The mixture was heated to 70 ℃ and stirred for 6 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) to give a white solid (45mg, 65.5%).

MS(ESI,pos.ion)m/z:475.00(M+1)；

¹H NMR(400MHz,CDCl₃):7.48(dd,J＝6.7,1.7Hz,1H),7.42–7.32(m,2H),7.05(s,1H),6.92(d,J＝4.0Hz,1H),6.62(t,J＝9.0Hz,2H),5.10(s,2H),5.07–4.97(m,1H),4.66(t,J＝9.2Hz,1H),4.27(dd,J＝9.6,6.4Hz,1H),3.96(ddd,J＝14.6,6.6,3.5Hz,1H),3.74(dt,J＝14.7,6.3Hz,1H)。

Example 10: (S) -5-chloro-N- ((3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1 (2- (methoxycarbonyl) -4-methylthiophen-3-yl) diazo tetrafluoroborate

To a stirred 6M aqueous hydrochloric acid solution (25mL) was slowly added methyl 3-amino-4-methylthiophene-2-carboxylate (10.27g,60.00 mmol). The mixture was stirred at room temperature for 30 minutes, cooled to 0 ℃ and diazotized by addition of a solution of sodium nitrite (4.2g,60mmol) in water (10 mL). The resulting diazonium salt is stirred at 0 ℃ for a further 1 hour. To this was added 60% aqueous fluoroboric acid solution (9mL) at 0 ℃ in one portion, and stirring was continued for 30 minutes. The filtrate was washed with cold water (50mL) and a mixed solvent of methanol and ether (50mL, (methanol/ether (v/v) ═ 1/4)) in this order, and dried to obtain a white solid (13.8g, 80.4%). The product was used directly in the next step.

Step 2 3-fluoro-4-methylthiophene-2-carboxylic acid methyl ester

To a round bottom flask were added (2- (methoxycarbonyl) -4-methylthiophen-3-yl) diazotetrafluoroborate (13.8g,48.3mmol) and p-xylene (100 mL). The mixture was heated to 160 ℃ and stirred for 8 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (petroleum ether/dichloromethane (v/v) ═ 5/1) to give a pale yellow oil (3g, 35.7%).

GC-MS:M＝174.0。

Step 3-4- (bromomethyl) -3-fluorothiophene-2-carboxylic acid methyl ester

To a round bottom flask was added methyl 3-fluoro-4-methylthiophene-2-carboxylate (3.00g,17.2mmol), N-bromosuccinimide (3.98g,22.4mmol), benzoyl peroxide (834mg,3.44mmol), and carbon tetrachloride (40 mL). The mixture was heated to 85 ℃ and stirred for 8 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10/1) to give a yellow oil (4g, 91.8%).

MS(ESI,pos.ion)m/z:252.90(M+1)。

Step 4 methyl 3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylate

To a round bottom flask was added 4- (bromomethyl) -3-fluorothiophene-2-carboxylic acid methyl ester (4.35g,17.2mmol), pyridin-2 (1H) -one (1.63g,17.2mmol), potassium carbonate (7.20g,51.6mmol), potassium iodide (571mg,3.44mmol), and acetone (50 mL). The mixture was heated to 60 ℃ and stirred for 4 hours. Filtration was carried out, the solvent was evaporated from the filtrate under reduced pressure, and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2/1) to give a yellow oil (700mg, 20%).

MS(ESI,pos.ion)m/z:268.00(M+1)。

Step 5 3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylic acid

To a solution of methyl 3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylate (450mg,1.7mmol) in ethanol (10mL) was added 1M aqueous sodium hydroxide (10 mL). Heated to 100 ℃ and stirred for 2 hours. The solvent was evaporated under reduced pressure and acidified to pH 1 by addition of 12M hydrochloric acid solution. Filtration, filter residue washing with water (10mL), oven drying to obtain white solid (400mg, 90%). The product was used directly in the next step. Step 6 benzyl (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) carbamate

To a solution of 3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophene-2-carboxylic acid (400mg,1.58mmol), benzyl alcohol (256.3mg,2.370mmol), and N, N-diisopropylethylamine (428.8mg,3.318mmol) in toluene (20mL) was added diphenyl phosphorazidate (695.6mg,2.528mmol), and the mixture was heated to 110 deg.C and stirred for 5.5 hours. After cooling to room temperature, water (30mL) was added, extraction was performed with ethyl acetate (40 mL. times.3), and the organic phases were combined, washed with water (30 mL. times.3) and saturated brine (50mL) in this order, and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2/1) gave a yellow solid (377.7mg, 66.7%).

MS(ESI,pos.ion)m/z:359.05(M+1)。

Step 7 (R) -3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -5- (hydroxymethyl) oxazolidin-2-one

To a solution of benzyl 3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) carbamate (377.7mg,1.054mmol) in dry tetrahydrofuran (10mL) under nitrogen protection at-78 ℃ was slowly added dropwise a solution of 1M lithium bistrimethylsilylamide in tetrahydrofuran (3mL,3 mmol). The mixture was stirred at-78 ℃ for 0.5 h, then warmed to 0 ℃ and stirred for 15min, then a solution of (R) -oxiran-2-ylmethylbutyrate (303.9mg,2.108mmol) in tetrahydrofuran (4mL) was slowly added dropwise, cooled to-78 ℃ and stirred for 2h, warmed to room temperature and stirred overnight. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/2) to give a white solid (300mg, 90%).

MS(ESI,pos.ion)m/z:325.10(M+1)。

Step 8 (R) - (3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl methanesulfonate

To a round bottom flask was added (R) -3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -5- (hydroxymethyl) oxazolidin-2-one (332mg,1.024mmol), N-diisopropylethylamine (677 μ L,4.095mmol), and dichloromethane (10 mL). The mixture was cooled to 0 ℃ and then methanesulfonyl chloride (158. mu.L, 2.047mmol) was slowly added dropwise and stirred at room temperature for 4 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a white solid (350mg, 85%).

MS(ESI,pos.ion)m/z:403.05(M+1)。

Step 9 (R) -5- (azidomethyl) -3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (R) - (3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl methanesulfonate (350mg,0.8698mmol) in N, N-dimethylformamide (10mL) was added sodium azide (565.4mg,8.698mmol), and the mixture was heated to 70 ℃ and stirred for 12 hours. The solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a white solid (290mg, 95.5%).

Step 10 (S) -5- (aminomethyl) -3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a solution of (R) -5- (azidomethyl) -3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (290mg,0.8302mmol) in tetrahydrofuran (20mL) were added water (2mL) and triphenylphosphine (435.5mg,1.660mmol), the mixture was stirred overnight at room temperature, and the solvent was evaporated under reduced pressure. The crude product was purified by column chromatography (dichloromethane/methanol (v/v) ═ 10/1) to give a colourless oil (250mg, 93.1%). MS (ESI, pos. ion) M/z 324.00(M + 1).

Step 11 5-chlorothiophene-2-carbonyl chloride

To a round bottom flask was added 5-chlorothiophene-2-carboxylic acid (325.2mg,2mmol) and thionyl chloride (10 mL). The mixture was heated to 90 ℃ and stirred for 3 hours. The solvent was evaporated under reduced pressure to give a crude product which was used in the next step without purification.

Step 12 (S) -5-chloro-N- ((3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a round bottom flask was added (S) -5- (aminomethyl) -3- (3-fluoro-4- ((2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (250mg,0.7733mmol), N-diisopropylethylamine (399.8mg,3.093mmol) and dichloromethane (30 mL). The mixture was cooled to 0 ℃ and then a solution of 5-chlorothiophene-2-carbonyl chloride (362.1mg,2mmol) in dichloromethane (5mL) was added, warmed to room temperature and stirred for 12 hours. Dichloromethane (50mL) was added and the mixture was washed with 1M sodium hydroxide (20 mL. times.2). The organic phase was dried over anhydrous sodium sulfate, filtered, the solvent was evaporated under reduced pressure and the crude product was purified by column chromatography (ethyl acetate) to give a white solid (250mg, 73.5%).

MS(ESI,pos.ion)m/z:468.10(M+1)；

¹H NMR(600MHz,DMSO-d₆)8.95(t,J＝5.8Hz,1H),7.69(t,J＝5.2Hz,2H),7.43(ddd,J＝8.8,6.6,1.9Hz,1H),7.20(d,J＝4.0Hz,1H),7.04(d,J＝5.0Hz,1H),6.41(d,J＝9.1Hz,1H),6.31–6.21(m,1H),4.97(s,2H),4.92-4.86(m,1H),4.17(t,J＝8.6Hz,1H),3.92–3.82(m,1H),3.68–3.54(m,2H)；

¹⁹F NMR(376MHz,DMSO-d₆):-139.37(s)。

Example 11: (S) -5-chloro-N- ((3- (4- ((4-chloro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

Step 1- ((5-bromothien-3-yl) methyl) -4-chloropyridin-2 (1H) -one

To a solution of 4-chloropyridin-2-ol (650mg,5.0mmol) in acetone (20mL) was added potassium carbonate (1.4g,10mmol) and 2-bromo-4- (bromomethyl) thiophene (1.3g,5.0 mmol). The mixture was heated to 65 ℃ and stirred for 2.5 hours, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. Water (25mL) was added thereto, and the mixture was extracted with ethyl acetate (20 mL. times.4). The organic phases were combined, washed with saturated brine (40mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 4/1) gave a white solid (1.20g, 77.6%).

MS(ESI,pos.ion)m/z:305.90(M+1)。

Step 2 (S) -2- ((3- (4- ((4-chloro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione

To the sealed tube were added in order 1- ((5-bromothien-3-yl) methyl) -4-chloropyridin-2 (1H) -one (870mg,2.9mmol), (R) -2- ((2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (700mg,2.9mmol), N' -dimethylethylenediamine (100mg,1.1mmol), cuprous iodide (110mg,0.57mmol), potassium carbonate (1.2g,8.6mmol), and 1, 4-dioxane (10 mL). The mixture was heated to 120 ℃ under nitrogen and stirred for 8 hours. Cooling to room temperature, vacuum filtering, and vacuum evaporating the filtrate to remove solvent. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2/3) to give a pale yellow solid (910mg, 68%).

MS(ESI,pos.ion)m/z:470.00(M+1)。

Step 3 (S) -5- (aminomethyl) -3- (4- ((4-chloro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one

To a suspension of (S) -2- ((3- (4- ((4-chloro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (910mg,1.9mmol) in ethanol (20mL) was added 40% aqueous methylamine solution (2.0 mL). The mixture was heated to 95 ℃ and stirred for 1.5 hours, and the solvent was evaporated under reduced pressure. The crude product was used in the next step without purification.

Step 4 (S) -5-chloro-N- ((3- (4- ((4-chloro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) thiophene-2-carboxamide

To a solution of (S) -5- (aminomethyl) -3- (4- ((4-chloro-2-oxopyridin-1 (2H) -yl) methyl) thiophen-2-yl) oxazolidin-2-one (646mg,1.90mmol) and 5-chlorothiophene-2-carboxylic acid (371mg,2.28mmol) in dichloromethane (40mL) was added HATU (1.08g,2.85mmol) and N, N-diisopropylethylamine (491mg,3.80 mmol). The mixture was stirred at room temperature for 6 hours. The solvent was evaporated under reduced pressure, methylene chloride (40mL) was added, and the organic phase was washed successively with saturated sodium hydrogencarbonate (30mL), saturated brine (30mL) and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and column chromatography of the crude product (dichloromethane/methanol (v/v) ═ 50/1) gave a pale yellow solid (310mg, 34%).

MS(ESI,pos.ion)m/z:484.00(M+1)；

1H NMR(400MHz,DMSO-d₆):8.93(t,J＝5.5Hz,1H),7.83(d,J＝7.3Hz,1H),7.67(d,J＝4.0Hz,1H),7.18(d,J＝4.0Hz,1H),6.96(s,1H),6.57(d,J＝1.9Hz,1H),6.49(s,1H),6.38(dd,J＝7.3,2.1Hz,1H),4.97(s,2H),4.92–4.83(m,1H),4.11(t,J＝8.9Hz,1H),3.78(dd,J＝8.5,6.7Hz,1H),3.60(t,J＝5.4Hz,2H)。

Example 12: (S) -N- ((3- (4- ((1H-pyrazol-1-yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) -5-chlorothiophene-2-carboxamide

Step 1- ((5-bromothien-3-yl) methyl) -1H-pyrazole

Pyrazole (1.00g,14.7mmol), 2-bromo-4- (bromomethyl) thiophene (3.76g,14.7mmol), cesium fluoride (4.67g,30.7mmol), and acetonitrile (50mL) were added sequentially to a 100mL two-necked round-bottomed flask, and the mixture was heated to reflux and stirred for 24 hours. After cooling to room temperature, water (100mL) was added and extracted with ethyl acetate (50 mL. times.3). The organic phases were combined, washed successively with water (50 mL. times.2) and saturated brine (50mL), and dried over anhydrous sodium sulfate. Filtration and evaporation of the solvent under reduced pressure followed by column chromatography of the crude product (petroleum ether/ethyl acetate (v/v) ═ 10/1) gave a yellow oil (1.32g, 37.0%).

MS(ESI,pos.ion)m/z:243.0(M+1)。

Step 2 (S) -2- ((3- (4- ((1H-pyrazol-1-yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione

Under nitrogen protection, (R) -2- ((2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione, 1- ((5-bromothien-3-yl) methyl) -1H-pyrazole (1.32g,5.43mmol), potassium carbonate (2.25g,16.3mmol), cuprous iodide (207mg,1.08mmol), N¹,N²Dimethylethane-1, 2-diamine (193mg,2.19mmol) and 1, 4-dioxane (20mL), and the mixture was heated to 120 ℃ and stirred for 10 hours. Cooled to room temperature, and the mixture was extracted with ethyl acetate (50 mL. times.3). MergingThe organic phase was washed with water (50 mL. times.2) and saturated brine (40mL) in this order, and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 1/1) gave a yellow oil (1.09g, 49.2%).

MS(ESI,pos.ion)m/z:409.2(M+1)。

Step 3 (S) -3- (4- ((1H-pyrazol-1-yl) methyl) thiophen-2-yl) -5- (aminomethyl) oxazolidin-2-one

Methylamine (40% aqueous solution, 2.7mL) was added to a solution of (S) -2- ((3- (4- ((1H-pyrazol-1-yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) isoindoline-1, 3-dione (1.09g,2.67mmol) in ethanol (30mL), heated to 95 ℃ and stirred for 1.5 hours. The solvent was evaporated under reduced pressure to give a white solid (0.743g, 100%). The crude product is directly put into the next step.

Step 4 (S) -N- ((3- (4- ((1H-pyrazol-1-yl) methyl) thiophen-2-yl) -2-oxooxazolidin-5-yl) methyl) -5-chlorothiophene-2-carboxamide

To a 50mL two-necked round bottom flask were added 2-chlorothiophene-5-carboxylic acid (0.52g,3.2mmol), N-diisopropylethylamine (1.84mL,10.7mmol), dichloromethane (30mL), HATU (1.52g,4.0mmol) in that order, and after stirring at room temperature for 1 hour, (S) -3- (4- ((1H-pyrazol-1-yl) methyl) thiophen-2-yl) -5- (aminomethyl) oxazolidin-2-one (0.742g,2.67mmol) was added. The mixture was stirred at room temperature for 6 hours, and extracted with dichloromethane (30 mL. times.2). The organic phases were combined, washed successively with water (20 mL. times.2) and saturated brine (20mL), and dried over anhydrous sodium sulfate. Filtration, evaporation of the solvent under reduced pressure and purification of the crude product by column chromatography (petroleum ether/ethyl acetate (v/v) ═ 4/1) gave a white solid (0.568g, 50.4%). MS (ESI, pos.ion) M/z 423.0(M + 1);

¹H NMR(600MHz,CDCl₃):7.56(d,J＝1.5Hz,1H),7.41(d,J＝2.1Hz,1H),7.34(d,J＝4.0Hz,1H),6.91(d,J＝4.0Hz,1H),6.77(d,J＝0.7Hz,1H),6.73(t,J＝6.0Hz,1H),6.36(d,J＝1.6Hz,1H),6.30(t,J＝2.0Hz,1H),5.23(s,2H),4.97–4.87(m,J＝9.4,6.5,3.2Hz,1H),4.08(t,J＝9.0Hz,1H),3.90(ddd,J＝14.7,6.4,3.1Hz,1H),3.78(dd,J＝9.2,6.7Hz,1H),3.73(dt,J＝14.7,6.2Hz,1H)。

biological activity assay

Inhibition of human FXa enzyme

The enzymatic activity of human coagulation factor xa (FXa) is determined by conversion of a chromogenic substrate specific for FXa. In this regard, factor Xa cleaves p-nitroaniline from the chromogenic substrate. The assay was performed on microtiter plates as follows.

The test substances were dissolved in 10% dimethylsulfoxide at different concentrations, 5. mu.L of compound was mixed with 10. mu.L of human FXa (10nM in 50mM Tris, 150mM NaCl, pH 8.3), incubated for 15min at 25 ℃ in a constant temperature incubator, and after incubation, 5. mu.L of FXa chromogenic substrate (800. mu.M, sigma) was added and absorbance values were measured kinetically at 405nM at 25 ℃. The test mixtures containing the test substances are compared with the control mixtures without test substances and the IC is calculated from these data₅₀The value is obtained.

In vitro anticoagulation assay

Compound for prolonging coagulation time of rabbit plasma

1. Preparation of Compounds at various concentrations

mu.L of each compound working solution (100mM) was diluted with dimethyl sulfoxide solution to give working solutions of respective concentrations.

2. Preparation of plasma samples

Taking a plurality of rabbits, carrying out intravenous injection of 3% pentobarbital solution (30mg/kg) on ear edges for anesthesia, collecting blood to 2mL by using a vacuum blood collection tube abdominal aorta containing 0.2mL of 3.8% sodium citrate, collecting a plurality of tubes, reversing the upper part and the lower part and mixing the mixture evenly for a plurality of times, standing the mixture for 10min, centrifuging the mixture for 10min at 3000rpm, sucking blood plasma of each tube, mixing all the blood plasma into the same centrifuge tube, subpackaging each tube with 1.6mL, and rapidly placing the mixture into a refrigerator at minus 80 ℃ for storage and standby.

3. Sample adding and blood coagulation time determination PT and APTT

Prepare 1.5mL EP tubes, add 180. mu.l per tubeAn L plasma sample; respectively adding 4 μ L of corresponding concentration medicine into each blood vessel specimen, adding 4 μ L of dimethyl sulfoxide solution into control group, shaking, mixing, and incubating at 37 deg.C for 5 min; PT and APTT are measured by a Sysmex CA1500 full-automatic hemagglutination instrument; dose-response curves were plotted and fitted to calculate the concentration of test compound that doubles the Clotting Time (CT)₂)。

TABLE 2 inhibition of human FXa activity and in vitro anticoagulation of the Compounds

And (4) conclusion: the compound has better coagulation factor Xa inhibitory activity and the function of prolonging the coagulation time.

Solubility testing of Compounds

Water (10mL) was added to a15 mL conical tube and the sample was added with shaking until the sample stopped dissolving, and the mixture was shaken in a 37 ℃ thermostatic water bath for 24h at a shaking speed of 40 rpm. After shaking, the sample was filtered through a water-based microporous membrane (0.45 μm, Φ 13mm), the initial filtrate was discarded, the subsequent filtrate (500 μ L) was precisely removed, and acetonitrile-water (500 μ L, v/v ═ 60/40) as a diluent was added to the filtrate, and the mixture was mixed well to obtain a sample solution.

The sample solution (40. mu.L) was taken, subjected to HPLC detection, and the sample concentration and the compound solubility were calculated by the external standard one-point method.

TABLE 3 solubility of the Compounds in Water

Example numbering	Chemical combinationSolubility of substance (mg/mL)
		Example 1	0.14
Example 2	0.15
		Example 3	0.16
Example 4	0.13
		Example 5	0.09
Example 6	0.18
		Example 7	0.08
Example 8	0.25
		Example 9	0.13
Example 10	0.12
		Example 11	0.15
Example 12	0.23

And (4) conclusion: the series of compounds have better solubility.

In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.

Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.

Claims

1. A compound which is a compound of formula (I), or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of a compound of formula (I):

wherein,

n is 0 or 1; r¹Is H, -NH₂、-CN、F. Cl or Br;

R³is H, -NH₂-CN, F, Cl, Br or C₁-C₄An alkyl group;

x, Y and Z are each independently CR⁴Or N;

2. The compound according to claim 1, wherein,

R³is H, -NH₂CN, -F, Cl, Br, methyl, ethyl, n-propyl or isopropyl;

and, R²Is H, F, Cl, Br, -OH, -C (═ O) OH, -NH₂Methyl, ethyl, propyl, methoxy, ethoxy or propoxy.

3. The compound according to claim 1, which is a compound represented by the general formula (II), or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a racemate, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt, or a prodrug of the compound represented by the general formula (II):

wherein,

n is 0 or 1;

R¹is H, -NH₂-CN, F, Cl or Br;

x, Y and Z are each independently CR⁴Or N; and

4. The compound of claim 1, comprising one of the following,

5. A pharmaceutical composition comprising a compound of any one of claims 1-4, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or combination thereof.

6. Use of a compound according to any one of claims 1 to 4 or a pharmaceutical composition according to claim 5 in the manufacture of a medicament for the prevention, treatment or treatment of a thromboembolic disorder.

7. Use of a compound according to any one of claims 1 to 4 or a pharmaceutical composition according to claim 5 in the manufacture of a medicament for inhibiting factor Xa.