A β-lactamase inhibitor containing urea bicyclic compound, preparation method and use thereof

Abstract

The present invention belongs to the field of pharmaceutical chemistry technology, and specifically relates to a β-lactamase inhibitor containing a urea bicyclic compound, a preparation method thereof and its use. The present invention provides a class of β-lactamase inhibitors with a completely new structure, and its structural formula is shown in Formula I. When such molecules are used in combination with traditional β-lactam antibiotics, they can reverse the drug resistance problem of most bacteria to β-lactam antibiotics due to the expression of β-lactamase, and have a good application prospect in the clinical treatment of bacterial infections.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical chemistry, and in particular relates to a β-lactamase inhibitor containing a urea bicyclic compound, a preparation method thereof and a use thereof. Background Art

[0002] Since Fleming discovered penicillin in the 1920s, the invention and clinical application of antibiotics have profoundly changed human health care and quality of life, and greatly increased the average life expectancy of people in various countries. However, the resistance of pathogens to antibiotics in use has also been observed as a scientific fact for a long time. In 1967, penicillin-resistant Streptococcus pneumoniae isolates were reported; then in 1976, penicillinase-producing Neisseria gonorrhoeae isolates were discovered; extended-spectrum cephalosporins Cefotaxime was released in 1980, and three years later, extended-spectrum beta-lactamase-producing Escherichia coli isolates were identified. As time goes by and new varieties are continuously put into clinical use, especially with the expansion of the scale of antibiotic use, the occurrence of clinical pathogen resistance cases seems to be more and more rapid.

[0003] In biological terms, antibiotic resistance is the natural result of the interaction between the environmental selection pressure brought about by antibiotic use and the adaptive capacity of pathogens dominated by genetic mutability. Although the large-scale use of antibiotics is undoubtedly a key factor in the development and spread of antibiotic resistance into a problem and crisis, the latest research shows that resistance to antibiotic compounds first isolated from natural products is a natural phenomenon that has existed before their industrial production. The known resistance mechanisms generally include: ① Chemical alteration and inactivation of antibiotics by hydrolysis by β-lactamase as the main method; ② Modification of drug binding sites of penicillin binding proteins (PBPs); ③ Reduction of intracellular drug accumulation levels, such as active efflux pumps; ④ Loss of bacterial outer membrane pores, limiting drug influx; ⑤ Formation of resistant bacterial biofilms.

[0004] The hydrolytic degradation of β-lactam antibiotics by β-lactamases represents the most clinically relevant mechanism of pathogen resistance. The majority of resistant bacteria on the WHO and US CDC priority or urgent threat lists express β-lactamases as the primary mechanism. The target sites of β-lactam antibiotics are penicillin-binding proteins on the bacterial cell membrane, which are biochemically involved in the biosynthesis and self-degradation of bacterial cell wall peptidoglycan (PG), including DD-transpeptidases (TP), carboxypeptidases (DDcarboxy-peptidases), and endopeptidases (DD-endopeptidases). The β-lactam heterobicyclic nucleus of typical β-lactam antibiotics is structurally very similar to the D-alanyl-D-alanine (D-ala-D-ala) portion of the bacterial cell wall peptidoglycan tetrapeptide side chain, which is the normal substrate for PBPs. PBPs with normal number levels and types are necessary for bacteria to maintain normal cell morphology and function and complete cell division and morphogenesis in the cell cycle. β-lactam antibiotics can effectively interfere with the normal physiological cycle of cell wall biosynthesis and degradation through irreversible covalent binding with PBPs, and directly destroy the protective function of bacterial cell walls against osmotic shock, leading to bacterial cell disintegration and death. However, some Gram-negative bacteria and a few Gram-positive bacteria can produce β-lactamases with similar structures to PBPs. These enzymes also have ultra-high affinity for many β-lactam antibiotics, but can hydrolyze and destroy the four-membered β-lactam ring of the bound antibiotics to make cells resistant. Various types of β-lactamases that evolve at an astonishing rate have great molecular diversity. Currently, more than 7,000 different β-lactamase protein molecules are known worldwide. Many pathogenic bacterial cells can express multiple β-lactamases with different specificities. At the same time, some β-lactamases have evolved ultra-broad-spectrum hydrolytic activities (ESBLs, carbapenemases, etc.), thus constituting a pathway for acquiring multiple drug resistance (MDR, XDR, PDR, etc.).

[0005] A straightforward strategy to address the above resistance mechanisms is to develop small molecule inhibitors (BLIs) of β-lactamases and restore the antibacterial activity of resistant β-lactam antibiotics through combination therapy. For example, "CN104364254B heterobicyclic compounds as β-lactamase inhibitors" provides a heterobicyclic compound that, when used in combination with traditional β-lactam antibiotics, can reverse the resistance of most bacteria to β-lactam antibiotics due to the expression of β-lactamases.

[0006] However, the effects of the above-mentioned β-lactamase inhibitors still have room for further improvement, and thus it is necessary to design β-lactamase inhibitors with new molecular structures. Summary of the invention

[0007] In view of the problems of the prior art, the present invention provides a β-lactamase inhibitor containing a urea bicyclic compound, a preparation method and use thereof, with the aim of providing a new class of β-lactamase inhibitors.

[0008] The compound of formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, has the following structural formula:

[0009]

[0010] in,

[0011] A 1 Selected from -CO-;

[0012] A 2 , A 3 , A 4 are independently selected from -O-, -S-, -NR 4 -、-N=、-CR 5 R 6 -、-CO-、-CR 7 = or key;

[0013] R 1 Selected from

[0014] R 2 Selected from hydrogen, halogen, cyano, C l ~C 6 Alkyl, C 3 ~C 6 Cycloalkyl, C l ~C 6 Alkoxy, C l ~C 6 Alkylthio, -NR 8 R 9 ; wherein the above alkyl, alkoxy, alkylthio is substituted by 1-3 of the following groups: halogen, C l ~C 3 Alkoxy and hydroxyl groups;

[0015] R 3 Selected from hydrogen, C l ~C 12 Alkyl, C 2 ~C 12 The alkenyl group, C 2 ~C 12 Alkynyl, C3 ~C 10 Cycloalkyl, C 3 ~C 10 wherein the above alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycloalkyl, heterocycloalkenyl is substituted by 1 to 3 of the following groups: halogen, C l ~C 3 The alkoxy and hydroxyl groups;

[0016] R 4 Selected from hydrogen, C l ~C 6 Alkyl, C 2 ~C 6 The alkenyl group, C 2 ~C 6 Alkynyl, C 3 ~C 8 Cycloalkyl, C 3 ~C 8 wherein the above alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl is substituted by 1-3 of the following groups: halogen, C l ~C 3 The alkoxy and hydroxyl groups;

[0017] R 5 and R 6 are independently selected from H, C l ~C 6 Alkyl, C 2 ~C 6 The alkenyl group, C 2 ~C 6 Alkynyl, C 3 ~C 8 Cycloalkyl, C 3 ~C 8 Heterocycloalkyl, aryl, heteroaryl, halogen, cyano;

[0018] or R 5 and R 6 The carbon atom to which it is attached forms a C 3 ~C 7 cycloalkyl, 3-7 membered aryl, 3-7 membered heteroaryl, 3-7 membered heterocycloalkyl or 3-7 membered heterocycloalkenyl;

[0019] R 7 Selected from hydrogen, halogen, C l ~C 6 Alkyl, C l ~C 6 Alkoxy, C 3 ~C8 Cycloalkyl, C 3 ~C 8 Heterocycloalkyl, aryl, heteroaryl;

[0020] M is selected from hydrogen, metal ions or organic cations;

[0021] X 1 and X 2 are independently selected from H, halogen or cyano.

[0022] Preferably, R 2 Selected from hydrogen, halogen, cyano, C l ~C 6 Alkyl, C 3 ~C 6 of a cycloalkyl group.

[0023] Preferably, A 2 Selected from -O-, -S-, -NR 4 -or-N=;

[0024] A 3 Selected from -O-, -S-, -NR 4 -、-N=、-CR 5 R 6 -、-CO-、-CR 7 = or key;

[0025] A 4 Selected from -NR 4 -、-N=、-CR 5 R 6 -、-CO-、-CR 7 = or key.

[0026] Preferably, the structural formula of the compound is shown in any one of the following formula II to formula X:

[0027]

[0028] in,

[0029] R 1 Selected from

[0030] R 2 Selected from hydrogen, halogen, cyano, C l ~C 6 Alkyl, C 3 ~C 6 Cycloalkyl;

[0031] R 3 Selected from hydrogen, C l ~C 12 Alkyl, C 2~C 12 The alkenyl group, C 2 ~C 12 Alkynyl, C 3 ~C 10 Cycloalkyl, C 3 ~C 10 wherein the above alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycloalkyl, heterocycloalkenyl is substituted by 1 to 3 of the following groups: halogen, C l ~C 3 The alkoxy and hydroxyl groups;

[0032] R 4 , R 4a , R 4b , R 4c , R 4d , R 4e , R 4f are independently selected from hydrogen, C l ~C 6 Alkyl, C 2 ~C 6 Alkenyl, C 2 ~C 6 Alkynyl, C 3 ~C 8 Cycloalkyl, C 3 ~C 8 The above alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl are optionally substituted by 1-3 of the following groups: halogen, C l ~C 3 The alkoxy and hydroxyl groups;

[0033] R 5 and R 6 are independently selected from H, C l ~C 6 Alkyl, C 2 ~C 6 The alkenyl group, C 2 ~C 6 Alkynyl, C 3 ~C 8 Cycloalkyl, C 3 ~C 8 Heterocycloalkyl, 5-6 membered aryl, 5-6 membered heteroaryl, halogen, cyano;

[0034] or R 5 and R 6 The carbon atom to which it is attached forms a C 3 ~C 7cycloalkyl, 3-7 membered aryl, 3-7 membered heteroaryl, 3-7 membered heterocycloalkyl or 3-7 membered heterocycloalkenyl;

[0035] R 7 Selected from hydrogen, halogen, C l ~C 6 Alkyl, C l ~C 6 Alkoxy, C 3 ~C 8 Cycloalkyl, C 3 ~C 8 Heterocycloalkyl, aryl, heteroaryl;

[0036] M is selected from hydrogen, metal ions or organic cations;

[0037] X 1 and X 2 are independently selected from H, halogen or cyano.

[0038] Preferably, R 5 and R 6 are independently selected from H, C l ~C 6 Alkyl, C 2 ~C 6 The alkenyl group, C 2 ~C 6 Alkynyl, C 3 ~C 8 Cycloalkyl, C 3 ~C 8 Heterocycloalkyl, 5-6 membered aryl, 5-6 membered heteroaryl, halogen, cyano;

[0039] or R 5 and R 6 The carbon atom to which it is attached forms a C 3 ~C 7 cycloalkyl, 3-7 membered aryl, 3-7 membered heteroaryl, 3-7 membered heterocycloalkyl or 3-7 membered heterocycloalkenyl;

[0040] R 7 Selected from hydrogen, halogen, C l ~C 6 Alkyl, C l ~C 6 Alkoxy, C 3 ~C 8 Cycloalkyl, C 3 ~C 8 The invention also includes a heterocycloalkyl group, a 5-6 membered aryl group, a 5-6 membered heteroaryl group.

[0041] Preferably, R 1 Selected from

[0042] Preferably, the structural formula of the compound is:

[0043]

[0044]

[0045] The present invention also provides a method for preparing the above compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof, comprising the following steps:

[0046]

[0047] Step 1, using compound MI as a raw material to carry out a cyclization reaction to obtain intermediate A or intermediate B;

[0048] Step 2, using intermediate A or intermediate B as a raw material to carry out a cyclization reaction to obtain a product;

[0049] Among them, A 1 , A 2 , A 3 , A 4 , R 1 , R 2 , R 5 , R 6 As claimed in any one of claims 1 to 7;

[0050] X is selected from O or NH;

[0051] PG 1 and PG 2 are independently selected from hydrogen or an amine protecting group;

[0052] PG 3 and PG 4 are independently selected from hydrogen or a hydroxy protecting group;

[0053] PG 5 Selected from hydrogen, a hydroxyl protecting group or an amine protecting group.

[0054] Preferably, the preparation method of the compound MI comprises the following steps:

[0055]

[0056] It is prepared by using compound M-II as a raw material, undergoing olefin metathesis reaction, ring closure, and then reduction and Mitsunobu reaction;

[0057] Among them, R a1 and R a1 are independently selected from hydrogen or methyl.

[0058] The present invention also provides use of the above compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof in preparing a drug for treating bacterial infection.

[0059] Preferably, the drug is a drug for treating infection caused by drug-resistant bacteria expressing β-lactamase.

[0060] The present invention also provides a drug for treating bacterial infection, which is a preparation prepared by using the above compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a solvate thereof as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0061] Preferably, the drug is a drug for treating infection caused by drug-resistant bacteria expressing β-lactamase.

[0062] The present invention also provides a combined drug for treating bacterial infection, which comprises the above drug and at least one β-lactam antibiotic.

[0063] The present invention also provides a compound preparation for treating bacterial infection, which is a compound preparation prepared by taking the above-mentioned combined drug as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0064] The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0065] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

[0066] "Substitution" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0067] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a -C b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C 1 -C 4 "Alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0068] "Alkyl" refers to a saturated hydrocarbon chain having the specified number of member atoms. 1 -C 6Alkyl refers to an alkyl group having 1 to 6 member atoms, for example 1 to 4 member atoms. The alkyl group can be straight or branched. Representative branched alkyl groups have one, two or three branches. The alkyl group may be optionally substituted with one or more substituents as defined herein. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl. The alkyl group may also be part of other groups, such as C 1 -C 6 Alkoxy.

[0069] "Cycloalkyl" refers to a saturated or partially saturated cyclic group having 3 to 14 carbon atoms and no ring heteroatoms and having a single ring or multiple rings (including fused, bridged and spirocyclic systems). For polycyclic systems with aromatic and non-aromatic rings without ring heteroatoms, when the point of attachment is located at a non-aromatic carbon atom, the term "cycloalkyl" (e.g., 5,6,7,8,-tetrahydronaphthalene-5-yl) is applicable. The term "cycloalkyl" includes cycloalkenyl groups, such as cyclohexenyl. Examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl and cyclohexenyl.

[0070] "Halogen" is fluorine, chlorine, bromine or iodine.

[0071] "Heterocycle" and "heterocycloalkyl" refer to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom;

[0072] "Heteroaryl" refers to an aromatic unsaturated ring group containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, an oxygen atom, or a sulfur atom;

[0073] "Alkenyl" is used to refer to a straight or branched hydrocarbon group containing at least one olefinic bond. 2 ~C 12 The alkenyl group refers to a straight or branched hydrocarbon group containing 2 to 12 carbon atoms and at least one olefinic bond. 2 ~C 4 The alkenyl group refers to a straight or branched hydrocarbon group containing 2 to 12 carbon atoms and at least one olefinic bond.

[0074] "Alkynyl" is used to refer to a straight or branched chain hydrocarbon group containing at least one acetylenic bond. 2 ~C 12 The alkynyl group refers to a straight-chain or branched hydrocarbon group containing 2 to 12 carbon atoms and at least one acetylenic bond. 2 ~C 4 The alkynyl group refers to a straight or branched hydrocarbon group containing 2 to 12 carbon atoms and at least one acetylenic bond.

[0075] "Cycloalkenyl" refers to a non-aromatic carbocyclic group containing at least one olefinic bond, including cyclized alkenyls. Cycloalkenyls may include bicyclic, polycyclic and spirocyclic systems. Examples of cycloalkenyls include, but are not limited to, cyclopentenyl and cyclohexenyl.

[0076] "Alkoxy" is used to indicate a saturated chain hydrocarbon group or a cyclic hydrocarbon group connected to a substituted atom via an oxygen atom. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, isopropoxy, tert-butoxy, cyclopropyloxy, and cyclobutyloxy.

[0077] "Alkylthio" is used to indicate a saturated chain hydrocarbon group or a cyclic hydrocarbon group connected to a substituted atom via a sulfur atom. Examples of alkoxy include, but are not limited to, methylthio, ethylthio, isopropylthio, tert-butylthio, cyclopropylthio, and cyclobutylthio.

[0078] "Stereoisomers" include enantiomers and diastereomers.

[0079] The hydrogen atoms in the compounds of the present invention may be various isotopes of hydrogen, such as protium (1H), deuterium (2H) or tritium ( 3 H).

[0080] The term "substituted" may mean substituted with one group or may mean substituted with at least two groups.

[0081] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients that constitute a pharmaceutical dosage form and physiologically compatible with the receptor.

[0082] The terms "salt" and "pharmaceutically acceptable salt" refer to acidic and / or basic salts of the above-mentioned compounds or their stereoisomers, formed with inorganic and / or organic acids and bases, and also include zwitterionic salts (inner salts), and also include quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final separation and purification of the compound. It can also be obtained by mixing the above-mentioned compound, or its stereoisomer, with a certain amount of acid or base appropriately (e.g., equivalent). These salts may form a precipitate in the solution and be collected by filtering, or be recovered after solvent evaporation, or be obtained by freeze drying after reaction in an aqueous medium. The salt described in the present invention can be a hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

[0083] In certain embodiments, one or more compounds of the present invention may be used in combination with each other. It is also possible to select the compounds of the present invention and any other active agent for use in combination to prepare a drug or pharmaceutical composition for regulating cell function or treating a disease. If a group of compounds is used, these compounds may be administered to a subject simultaneously, separately or sequentially.

[0084] The present invention provides a class of β-lactamase inhibitors with a novel structure. When used in combination with traditional β-lactam antibiotics, this class of molecules can reverse the drug resistance of most bacteria to β-lactam antibiotics due to the expression of β-lactamase, and has a good application prospect in the clinical treatment of bacterial infections.

[0085] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0086] The following is a further detailed description of the above contents of the present invention through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies implemented based on the above contents of the present invention belong to the scope of the present invention. DETAILED DESCRIPTION

[0087] The reagents and materials used in the following examples, unless otherwise specified, are all commercially available.

[0088] Example 1 Preparation of Intermediate M-15

[0089] Synthesis route:

[0090]

[0091] Step 1 Synthesis of Compound M-2:

[0092] Compound M-1 (7.0 g, 31.9 mmol, CAS No. 95715-85-8) and 15 mL of dichloromethane were added to a 50 mL three-necked flask. The reaction solution was cooled to 0 ° C in an ice bath. 2,2-dimethoxypropane (19 mL, 160 mmol) was added to the reaction solution under stirring. The solution was a colorless clear liquid. After stirring for 10 minutes, boron trifluoride ether (0.2 mL, 0.16 mmol) was added dropwise to the reaction solution. The solution was naturally warmed to room temperature and stirred overnight. The solution became a red clear liquid. As the stirring time increased, the color of the solution gradually darkened until it became black. After the reaction was completed by TLC monitoring, 2 mL of saturated sodium bicarbonate solution was added to the reaction solution and stirred for 30 minutes. The reaction solution was then concentrated to remove the solvent and excess 2,2-dimethoxypropane. 20 mL of ethyl acetate was added to the concentrated solution, washed with saturated brine, and the ethyl acetate phase was collected. Anhydrous NaSO 4 After drying and evaporating the solvent under reduced pressure, 7.7 g of black oily crude product M-2 was obtained with a yield of 93.0%, which was directly used for subsequent reactions.

[0093] 1 H NMR (400 MHz, CDCl 3 ): δ4.38(t,J=4.0Hz,1H), 4.14(d,J=4.0Hz,2H), 3.76(s,3H), 1.50(s,6H), 1.41(s,9H).

[0094] LCMS: theoretical value [M+H] + =259.14, measured value [M+H] + =260.1.

[0095] Step 2 Synthesis of Compound M-3:

[0096] Compound M-2 (4.0 g, 15.4 mmol) was added to a 100 mL four-necked round-bottom flask, and 40 mL of anhydrous tetrahydrofuran was added to dissolve the mixture, and the solution was a yellow clear liquid. The reaction flask was cooled to -78 ° C using a dry ice / ethanol bath, and mechanically stirred. Under nitrogen protection, a toluene solution of diisobutylaluminum hydride (1.5 M, 17.5 mL, 26.2 mmol) was added dropwise to the reaction solution through a constant pressure dropping funnel, and the reaction solution temperature was controlled below -65 ° C. After the addition was completed, the reaction was maintained at -78 ° C and stirred for about 4 hours. The reaction was monitored by TLC until complete.

[0097] 4 mL of methanol was slowly added dropwise to the reaction solution through a constant pressure dropping funnel to quench the reaction (note that gas is released during the dropwise addition), and the temperature in the reaction solution was maintained below -60 ° C. After the dropwise addition was completed, the temperature was naturally raised to room temperature under stirring, and then 50 mL of potassium sodium tartrate solution (13.0 g of potassium sodium tartrate tetrahydrate was dissolved in 50 mL of water) was added dropwise to the reaction solution. The organic phase was collected by a separatory funnel, and the aqueous phase was extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain 3.5 g of yellow oily crude product M-3, with a yield of 98.9%, which was directly used in subsequent reactions.

[0098] 1 H NMR (400 MHz, CDCl 3 ): δ9.54(s,1H), 4.31-4.11(m,1H), 4.02(d,J=8.0Hz,2H), 1.45(s,6H), 1.37(s,9H).

[0099] Step 3 Synthesis of Compound M-4:

[0100] Methyl triphenylphosphine bromide (16.4 g, 45.8 mmol) and 20 mL of anhydrous tetrahydrofuran were added to a 100 mL three-necked flask, and the solution was a white suspension. The reaction solution was cooled to 0°C under nitrogen protection, and a tetrahydrofuran solution of sodium hexamethyldisilazide (23.0 mL, 2M, 46.0 mmol) was added dropwise to the reaction solution, and the reaction solution was significantly heated and turned into a yellow suspension. After the addition was completed, the reaction solution was stirred at room temperature for 2 hours, and then the reaction bottle was cooled to -78°C with a dry ice / ethanol bath, and compound M-3 (3.50 g, 1.53 mmol) was dissolved in 7 mL of anhydrous tetrahydrofuran, and was added dropwise to the reaction solution through a constant pressure dropping funnel under nitrogen protection, and the temperature of the reaction solution was controlled below -65°C, and the solution was dripped in about 1 hour. After the addition was completed, the solution was stirred for 1 hour, and then naturally heated to room temperature and stirred for another hour. The reaction was monitored by TLC until it was complete.

[0101] 15 mL of saturated ammonium chloride solution was added dropwise to the reaction solution to quench the reaction, the organic phase was separated and collected, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a dark brown oily crude product. 30 mL of n-hexane was added to the crude product, cooled to 0°C and stirred, a white solid precipitated, the filtrate was collected by filtration, the filter cake was washed with a small amount of n-hexane, and the filtrate was concentrated to obtain 4.0 g (yield 115.3%) of brown oily crude product M-4, which was directly used in subsequent reactions.

[0102] Step 4 Synthesis of Compound M-5:

[0103] In a 50 mL three-necked round-bottom flask, the crude compound 4 (4.0 g, 17.6 mmol) was dissolved in 20 mL of acetonitrile. The solution was yellow and clear. Then copper chloride dihydrate (3.0 g, 17.6 mmol) was added all at once. The solution became a green suspension. The mixture was stirred at room temperature for 1 hour. The reaction was monitored by TLC until completion.

[0104] The reaction mixture was filtered under reduced pressure to remove insoluble solids, and the filtrate was concentrated. 4 mL of ethyl acetate was added to the concentrate to dilute it, and then aqueous ammonia (1 mL) was added and stirred. The collected organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a brown oily crude product. The product was purified by silica gel chromatography to obtain 1.0 g of a yellow oil. The total yield of compound 3 to compound 5 in three steps was 34.7%.

[0105] 1 H NMR (400 MHz, CDCl 3 ): δ5.89-5.76(m,1H), 5.25(d,J=12.0Hz,2H), 4.97(brs,1H), 4.31-4.17(m,1H), 3.76-3.58(m,2H), 2.50(brs,1H), 1.46(s,9H).

[0106] Step 5 Synthesis of Compound M-6:

[0107] Compound M-5 (1.0 g, 2.35 mmol) was dissolved in 10 mL of N,N-dimethylformamide in a 50 mL three-necked bottle. The solution was yellow and clear. Imidazole (336 mg, 4.93 mmol) was added and stirred for 15 minutes. The solution was yellow and clear. Tert-butyldiphenylsilyl chloride (646 mg, 2.35 mmol) was then added dropwise to the reaction solution. After the addition was complete, the solution was yellow and clear. Stir at room temperature overnight. The reaction was monitored by TLC until the reaction was complete.

[0108] Add 30 mL of ice water and 5 mL of ethyl acetate to the reaction solution, stir for 10 minutes, collect the organic phase, and extract the aqueous phase once with 5 mL of ethyl acetate. The combined organic phase is washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude yellow oily product. Purify by silica gel chromatography to obtain 1.2 g of a yellow oily product with a yield of 96.0%.

[0109] 1 H NMR (400 MHz, CDCl 3): δ7.56(d,J=8.0Hz, 4H), 7.40-7.26(m,6H), 5.82-5.71(m,1H), 5.10(d,J=8.0Hz, 2 H), 4.75 (brs, 1H), 4.15 (brs, 1H), 3.57 (d, J = 4.0Hz, 2H), 1.38 (s, 9H), 0.98 (s, 9H).

[0110] LCMS: theoretical value [M-Boc+H] + =326.2, measured value [M-Boc+H] + =326.3.

[0111] Step 6 Synthesis of Compound M-7:

[0112] Potassium carbonate (1.34 g, 9.6 mmol), potassium ferrocyanide (3.18 g, 9.6 mmol) and methanesulfonamide (0.31 g, 3.2 mmol) were added to a 50 mL three-necked flask in sequence, and then 10 mL of water and 8 mL of tert-butanol were added, stirred for 10 minutes, and then potassium osmate (25 mg) and (DHQD) were added. 2 PHAL (25 mg), stirred for 5 minutes. Then compound M-6 (1.37 g, 3.2 mmol) dissolved in 2 mL of tert-butyl alcohol was added to the above reaction solution, and the reactant was a brown suspension. Stirred at room temperature overnight, and the reaction was monitored by TLC until the reaction was complete.

[0113] After tert-butanol in the reaction solution was evaporated, the concentrate was extracted twice with ethyl acetate. The combined organic phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a brown oily crude product. The product was purified by silica gel chromatography to obtain 1.05 g of a yellow oily product with a yield of 71.0%.

[0114] 1 H NMR (400 MHz, CDCl 3 ): δ7.57(d,J=4.0Hz, 4H), 7.31-7.42(m,6H), 4.94(d,J=4.0Hz, 1H), 3.99-3.91(m,1H) ), 3.86-3.69(m,2H), 3.69-3.56(m,1H), 3.56-3.35(m,2H), 1.37(s,9H), 1.01(s,9H).

[0115] LCMS: theoretical value [M-Boc+H] + =360.24, measured value [M-Boc+H] + =360.1.

[0116] Step 7 Synthesis of Compound M-8:

[0117] In a 25 mL single-necked bottle, add compound M-7 (0.8 g, 1.74 mmol), dissolve in 9 mL of dichloromethane, add imidazole (142 mg, 2.09 mmol), stir at room temperature for 5 minutes, then add tert-butyldimethylsilyl chloride (315 mg, 2.09 mmol) to the reaction solution, stir at room temperature for about 6 hours, and monitor the reaction by TLC until the reaction is completed.

[0118] 10 mL of water was added to the reaction solution, and the mixture was stirred and allowed to stand. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined, washed with water and saturated brine, respectively, dried over anhydrous sodium sulfate, and concentrated to obtain a yellow oil. The mixture was purified by silica gel chromatography to obtain 845 mg of a colorless oil with a yield of 84.6%.

[0119] 1 H NMR (400 MHz, CDCl 3 ): δ7.66 (d, J = 8.0Hz, H), 7.47-7.34 (m, 6H), 5.16 (d, J = 4.0Hz, 1H), 4.03-3.92 (m, 1H), 3.8 2-3.51(m,5H), 2.90(d,J=4.0Hz,1H), 1.43(s,9H), 1.06(s,9H), 0.88(s,9H), 0.02(s,6H).

[0120] Step 8 Synthesis of Compound M-9:

[0121] In a 25 mL single-necked bottle, add compound M-8 (0.84 g, 1.46 mmol), dissolve in 8 mL of ethyl acetate, add diisopropylethylamine (0.95 g, 7.30 mmol) under stirring at room temperature, and then add a solution of sulfur trioxide pyridine (0.70 g, 4.39 mmol) dissolved in dimethyl sulfoxide (8 mL), stir at room temperature for 6 hours, and monitor the reaction by TLC until the reaction is completed.

[0122] The reaction solution was poured into a 50 mL beaker, 10 mL of ice water was added, stirred evenly and allowed to stand, the aqueous phase was extracted with ethyl acetate, the organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate to obtain a yellow oily crude product, which was purified by silica gel chromatography to obtain 0.56 g of a light yellow oily substance with a yield of 66.9%.

[0123] 1 H NMR (400 MHz, CDCl 3): δ7.55(d,J=4.0H, 4H), 7.41-7.29(m,6H), 5.50(d,J=8.0Hz, 1H), 4.72-4.64( m,1H), 4.33-3.85(m,4H), 1.40(s,9H), 0.98(s,9H), 0.82(s,9H), 0.02(s,6H).

[0124] Step 9 Synthesis of Compound M-10:

[0125] In a 50mL three-necked flask, add methyltriphenylphosphonium bromide (2.586g, 7.23mmol), add 24mL of anhydrous tetrahydrofuran, ventilate the reaction flask three times with a nitrogen balloon, add a tetrahydrofuran solution of sodium bis(trimethylsilyl)amide (3.6mL, 2M in THF, 7.2mmol) dropwise under stirring at room temperature, the solution becomes a bright yellow turbid liquid, stir at room temperature for 2 hours, add compound M-9 (1.38g, 2.4mmol) dissolved in 3mL of tetrahydrofuran dropwise to the above reaction solution, heat to 40°C after the addition is complete, and react for 3 hours. Monitor the reaction by TLC until it is complete.

[0126] The reaction solution was cooled to room temperature, and 6 mL of cold saturated ammonium chloride solution was added dropwise in an ice-water bath at a temperature of about 5°C to quench the reaction. The solution was allowed to stand and then separated. The aqueous phase was extracted twice with ethyl acetate, and the organic phases were combined, washed three times with saturated brine, dried over anhydrous sodium sulfate, and the organic solvent was evaporated to obtain a brown-yellow oily crude product.

[0127] 75 mL of petroleum ether was added to the crude product under stirring to precipitate a large amount of triphenylphosphine oxide solid. The product was filtered through a Buchner funnel and the filter cake was washed with a small amount of petroleum ether. The filtrate was concentrated to obtain a pale yellow oily crude product. The product was purified by silica gel chromatography to obtain 0.97 g of a colorless oily product with a yield of 70.3%.

[0128] 1 H NMR (400 MHz, DMSO-d 6 ): δ7.36(d,J=4.0Hz, 4H), 7.60-7.40(m,6H), 6.95(d,J=4.0Hz, 1H), 5.05(d,J=20.0Hz, 2H), 4.23-4 .11(m,1H), 4.02(s,2H), 3.63(d,J=4.0Hz,2H), 1.40(s,9H), 1.00(s,9H), 0.85(s,9H), 0.02(s,6H).

[0129] LCMS: theoretical value [M-Boc+1] + =569.34, measured value [M-Boc+1] + =470.3.

[0130] Step 10 Synthesis of compound M-11:

[0131] In a 100 mL single-necked bottle, add compound M-10 (1.82 g, 3.27 mmol), dissolve it with 36 mL of dichloromethane, add zinc bromide (1.84 g, 8.18 mmol) under stirring, stir and react at room temperature for 16 hours, and monitor the reaction by TLC until it is complete.

[0132] Add 25 mL of saturated sodium bicarbonate solution to the reaction solution under stirring, stir at room temperature for half an hour, stand and separate, extract the aqueous phase twice with dichloromethane, combine the organic phases, wash twice with saturated brine, dry over anhydrous sodium sulfate, and concentrate to obtain a crude product. Purify by silica gel chromatography to obtain 1.2 g of light yellow oil, with a yield of 80.4%.

[0133] 1 H NMR (400 MHz, DMSO-d 6 ): δ7.62(d,J=12.0Hz, 4H), 7.50-7.40(m,6H), 5.07(d,J=4.0Hz, 4H), 4.12(s,2H), 3.65-3 .57(m,1H), 3.47(d,J=8.0Hz,2H), 1.62(brs,2H), 1.00(s,9H), 0.85(s,9H), 0.02(s,6H).

[0134] LCMS: theoretical value [M+H] + =469.28, measured value [M+H] + =470.5.

[0135] Step 11 Synthesis of Compound M-12:

[0136] In a 50 mL single-mouth bottle, add compound M-11 (1.10 g, 2.35 mmol), dissolve it with 12 mL N,N-dimethylformamide, add diisopropylethylamine (0.45 g, 3.51 mmol) under stirring. Then add 2-bromo-N-methoxy-N-methylacetamide (0.51 g, 2.8 mmol) to the reaction solution at once, stir and react at room temperature for 6 hours, and monitor the reaction by TLC until completion.

[0137] Add 15 mL of water and 15 mL of ethyl acetate to the reaction solution, stir, let stand and then separate the liquids. The aqueous phase was extracted twice with ethyl acetate. The organic phases were combined, washed with 10 mL of water, washed twice with saturated brine, and dried over anhydrous sodium sulfate to obtain 1.2 g of a yellow oily crude product with a yield of 89.4%, which was directly used in the next reaction.

[0138] 1 H NMR (400 MHz, CDCl 3): δ7.67(d,J=4.0Hz, 4H), 7.42-7.32(m,6H), 5.17(d,J=4.0Hz, 2H), 4.03(d,J=4 .0Hz, 2H), 3.67-3.65(m,1H), 3.64(s,3H), 3.62-3.29(m,4H), 3.12(s,3H), 2.26 -1.90(m,1H), 1.07(s,9H), 0.86(s,9H), 0.02(s,6H).

[0139] LCMS: theoretical value [M+H] + =570.33, measured value [M+H] + =571.5.

[0140] Step 12 Synthesis of Compound M-13:

[0141] Compound M-12 (1.2 g, 2.1 mmol) was placed in a 50 mL single-mouth bottle, 12 mL of triethylamine was added, and then di-tert-butyl dicarbonate (0.92 g, 4.2 mmol) was added. After nitrogen replacement twice, the reaction was refluxed for 5 hours. The reaction was monitored by TLC until completion (developing solvent PE:EA=6:1, product Rf=0.65).

[0142] After the reaction solution was cooled to room temperature, it was directly concentrated to obtain a yellow oily crude product, which was purified by silica gel chromatography to obtain 1.02 g of a light yellow oily product with a yield of 72.4%.

[0143] 1 H NMR (400 MHz, CDCl 3 ): δ7.61 (d, J = 4.0Hz, 4H), 7.28-7.28 (m, 6H), 5.24 (d, J = 16.0Hz, 1H), 4.99 (d, J = 2.0Hz, 1H), 4.70 (dt, J = 64.0, 6Hz, 1H), 4.27 -3.74(m,6H), 3.56(s,3H), 3.05(s,3H), 1.38(s,9H), 0.99(s,9H), 0.84(s,9H), 0.02(s,6H).

[0144] LCMS: theoretical value [M-Boc+H] + =670.38, measured value [M-Boc+H] + =571.5.

[0145] Step 13 Synthesis of Compound M-14:

[0146] Compound M-13 (1.0 g, 1.49 mmol) was added to a 25 mL three-necked flask, dissolved with 10 mL of anhydrous tetrahydrofuran, replaced with nitrogen three times, and the temperature of the reaction solution was reduced to below 5°C in an ice-water bath. 5.2 mL of vinyl grignard reagent (1 M in THF, 5.22 mmol) was added dropwise with a dropping funnel. The addition was completed in about 10 minutes. The reaction was reacted in an ice-water bath for 2 hours, and the reaction was monitored by TLC until completion.

[0147] The reaction solution was poured into 10% citric acid solution (10 mL) with stirring below 0°C to quench the reaction and stirred at room temperature for 1 hour. The reaction solution was allowed to stand and then separated. The aqueous phase was extracted twice with ethyl acetate. The organic phases were combined, washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow oily crude product, which was purified by silica gel chromatography to obtain 0.56 g of a light yellow oil (yield 58.9%).

[0148] 1 H NMR (400 MHz, CDCl 3 ): δ7.61(d,J=6.0Hz, 4H), 7.43-7.28(m,6H), 6.42(t,J=4.0Hz, 1H), 5.90(dd,J=4.0, 17Hz, 2H), 5.10(dd,J =4.0, 20Hz, 2H), 4.86-4.61(m,1H), 4.18-3.70(m,6H), 1.35(s,9H), 1.01(s,9H), 0.86(s,9H), 0.02(s,6H).

[0149] LCMS: theoretical value [M-Boc+H] + =637.36, measured value [M-Boc+H] + =538.6.

[0150] Step 14: Synthesis of Compound M-15:

[0151] Compound M-14 (0.49 g, 0.768 mmol) was placed in a 25 mL three-necked flask, 10 mL of toluene was added, the reaction flask was fully ventilated with nitrogen three times, 25 mg of the second-generation Grubbs reagent (0.039 mmol, CAS No: 301224-40-8) was quickly added to the reaction solution, the reaction flask was ventilated with nitrogen twice again, the reaction was carried out at 80 ° C for 5 hours, and the reaction was monitored by TLC until completion.

[0152] The reaction solution was concentrated to obtain a brown oil, which was purified by silica gel chromatography to obtain 0.37 g of a light yellow oil with a yield of 78.7%. After standing and cooling, it became a light yellow crystalline solid.

[0153] 1H NMR (400 MHz, CDCl 3 ): δ7.54(d,J=6.0Hz,4H), 7.43-7.30(m,6H), 6.31(s,1H), 4.55(s,2H), 4.20(s,2H), 3.92( d,J=8.0Hz, 2H), 3.67(t,J=4.0Hz, 1H), 1.44(s,9H), 0.96(s,9H), 0.85(s,9H), 0.02(s,6H).

[0154] LCMS: theoretical value [M-Boc+H] + =609.33, measured value [M-Boc+H] + =510.4.

[0155] Example 1 Synthesis of Compound 1

[0156]

[0157] Step a Synthesis of Compound 1A

[0158] Compound M-15 (2.26 g, 3.71 mmol) was dissolved in ethanol (40 mL) in a 100 mL single-necked bottle, and the reaction bottle was placed in an ice bath. P-toluenesulfonic acid (0.19 g, 1.11 mmol) was added to the reaction solution under stirring. The reaction solution was yellow and clear, and stirred at room temperature overnight. TLC monitored the reaction until it was complete, and saturated sodium bicarbonate solution (20 mL) was added to the reaction solution to quench the reaction. Ethanol was evaporated under reduced pressure at a bath temperature of 35°C, and 20 mL of saturated sodium bicarbonate solution was added to the residual liquid. Ethyl acetate (20 mL×3) was used for extraction, and the organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a yellow oily crude product. 0.75 g of yellow oily product 1A was obtained by column chromatography purification, with a yield of 40.8%. ESI-MS: [M+H] + =496.4.

[0159] Step b Synthesis of Compound 1B

[0160] At 0°C, DIAD (0.65 g, 3.23 mmol) was slowly added dropwise to a solution of compound 1A (1.00 g, 2.01 mmol), phthalimide (0.40 g, 2.69 mmol) and triphenylphosphine (0.85 g, 3.23 mmol) in THF (10 ml). After the addition was complete, the reaction mixture was purged with nitrogen. Stirring at ambient temperature for 2 hours, TLC and LC-MS showed that the raw material was completely consumed. The reaction mixture was concentrated to dryness and purified by column chromatography (10-30% ethyl acetate in hexane) to obtain compound 1B as a colorless transparent liquid (1.12 g, yield 89.6%).

[0161] ESI-MS: [M+H + ]=625.2.

[0162] Step c: Synthesis of Compound 1C

[0163] At 0°C, NaBH was slowly added to a solution of compound 1B (1.10 g, 1.79 mmol) and cerium trichloride (0.54 g, 2.20 mmol) in methanol (10 ml). 4 (0.025 g, 0.66 mmol). After the reaction mixture was stirred at ambient temperature for 30 minutes, TLC showed that the starting material was completely consumed. Saturated aqueous ammonium chloride solution was added to the reaction mixture and extracted with ethyl acetate. The organic phase was dried, filtered and concentrated. The residue was purified by column chromatography (20-40% ethyl acetate in hexane) to obtain compound 1C as a colorless transparent liquid (1.01 g, yield 90.0%).

[0164] ESI-MS: [M+H + ]=627.1.

[0165] Step d: Synthesis of Compound 1D

[0166] At 0°C, DIAD (0.48 g, 2.39 mmol) was slowly added dropwise to a DCM (10 ml) solution of compound 1C (1.00 g, 1.61 mmol), N-benzyloxy-o-nitrosulfonamide (0.55 g, 1.79 mmol) and triphenylphosphine (0.63 g, 2.39 mmol). After the addition was completed, the reaction mixture was purged with nitrogen. Stirring at ambient temperature for 2 hours, TLC and LC-MS showed that the raw material was completely consumed. The reaction mixture was concentrated to dryness and purified by column chromatography (20-40% ethyl acetate in hexane) to obtain compound 1D as a colorless transparent liquid (1.37 g, yield 92.7%). ESI-MS: [M+H + ]=917.2.

[0167] Step e: Synthesis of Compound 1E

[0168] To a solution of compound 1D (1.37 g, 1.49 mmol) in ethanol (12 ml) was added hydrazine hydrate (0.54 g, 8.65 mmol, 80%) at ambient temperature. After the mixture was refluxed at 80°C for 2 hours, TLC showed that the starting material was completely consumed. The reaction mixture was concentrated, the residue was diluted with water, extracted with ethyl acetate, and the organic phase was dried, filtered, and concentrated without further purification.

[0169] ESI-MS: [M+H + ]=663.2.

[0170] The concentrate from the previous step was dissolved in DCM (10 ml), and triethylamine (0.35 g, 3.46 mmol) and di-tert-butyl dicarbonate (0.76 g, 3.46 mmol) were added in sequence at 0°C. The reaction mixture was stirred at ambient temperature for 2 hours, and TLC showed that the starting material was completely consumed. The reaction mixture was concentrated, and the residue was redissolved in THF (10 ml), and TBAF (5.19 ml, 5.19 mmol, 1M in THF) was added. Stirring at ambient temperature for 1 hour, TLC showed that the starting material was completely consumed. The reaction mixture was concentrated, the residue was diluted with water, extracted with ethyl acetate, and the organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (20-40% ethyl acetate in hexane) to give compound 1E as a colorless transparent liquid (0.72 g, three-step yield 64.2%). ESI-MS: [M+H + ]=649.2.

[0171] Step f: Synthesis of Compound 1F

[0172] To a solution of periodic acid (0.62 g, 2.73 mmol) in wet acetonitrile (9 ml, containing 0.75% water) was added chromic anhydride (6 mg, 0.06 mmol) at ambient temperature and the mixture was stirred until complete dissolution was achieved.

[0173] At 0°C, the freshly prepared periodic acid / chromic anhydride solution was slowly added dropwise to a solution of compound 1E (0.72 g, 0.91 mmol) in wet acetonitrile (10 ml, containing 0.75% water). After the addition was complete, the reaction mixture was stirred at ambient temperature for 30 minutes. LC-MS showed that the starting material was completely consumed. Dilute with water, extract with ethyl acetate, and dry the organic phase, filter, and concentrate without further purification. ESI-MS: [M+H + ]=645.1.

[0174] The concentrate from the previous step was dissolved in a mixed solvent of TFA / DCM (8 ml, 1 / 5, v / v). Stirred at ambient temperature for 1 hour, LC-MS showed that the reaction was complete. The reaction solution was concentrated, diluted with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic phase was dried, filtered, and concentrated. The residue was purified by column chromatography (3-10% methanol in dichloromethane) to obtain compound 1F as a colorless transparent liquid (0.15 g, two-step yield 37.1%). ESI-MS: [M+H + ]=445.0.

[0175] Step g: Synthesis of Compound 1G

[0176] To a solution of compound 1F (0.15 g, 0.34 mmol) and potassium carbonate (0.28 g, 2.04 mmol) in methanol (5 ml) was added thioglycolic acid (0.16 g, 1.70 mmol). The reaction mixture was stirred overnight at ambient temperature, and TLC showed that the starting material was completely consumed. The reaction mixture was concentrated, re-added with a mixed solvent of DCM / MeOH (10 / 1, v / v) to dissolve, and filtered. The filtrate was concentrated and purified by column chromatography (3-10% methanol in dichloromethane) to obtain compound 1G as a white solid (0.045 g, yield 51.4%).

[0177] 1 H NMR (400 MHz, DMSO-d 6 )δ7.84(s,1H),7.38-7.23(m,5H),6.59(d,J=6.6Hz,1H),4.84-4.71(m,1H),4.64(s,2H),3.63(s,2H) ,3.28-3.15(m,2H),2.90-2.76(m,1H),2.43-2.29(m,1H),2.05(dd,J=17.1,7.4Hz,1H).ESI-MS:[M+H + ]=260.0.

[0178] Step h: Synthesis of compound 1H

[0179] At 0°C, a solution of triphosgene (0.014 g, 0.05 mmol) in acetonitrile (0.5 ml) was slowly added dropwise to a solution of compound 1G (0.03 g, 0.12 mmol) and DIPEA (0.05 g, 0.39 mmol) in acetonitrile (6 ml) . After the addition was complete, the reaction mixture was stirred overnight at ambient temperature, and TLC showed that the starting material was completely consumed. The reaction mixture was concentrated to dryness, diluted with ethyl acetate, and washed twice with water and saturated brine. The organic phase was dried, filtered, concentrated, and purified by column chromatography (2-10% methanol in dichloromethane) to give compound 1H as a white solid (0.023 g, yield 69.7%). ESI-MS: [M+H + ]=286.1.

[0180] Step i: Synthesis of compound 1I

[0181] Pd / C (2 mg, 0.02 mmol) was added to a solution of compound 1H (0.02 g, 0.07 mmol) in methanol (4 ml). The reaction mixture was replaced twice with nitrogen and hydrogen respectively. After the reaction mixture was stirred overnight at ambient temperature under a hydrogen atmosphere, LC-MS showed that the starting material was completely consumed. The reaction mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness to obtain compound 1I as a light black solid without further purification. ESI-MS: [M+H + ]=196.1.

[0182] Step j: Synthesis of compound 1

[0183] To a solution of compound 1I (0.02 g, 0.1 mmol) in pyridine (3 ml) was added sulfur trioxide pyridine (80 mg, 0.5 mmol). After the reaction mixture was stirred at ambient temperature overnight, LC-MS showed complete consumption of the starting material. The suspension was concentrated (<30°C) to dryness and resuspended in DCM. The solid was filtered off through a 0.45u Nalgene filter and the filtrate was concentrated to obtain an orange oil. The crude residue was purified by silica gel chromatography (0-30% methanol in dichloromethane), the product was dissolved in 5 mL of purified water and passed through a sodium ion exchange resin, washed with purified water, and the filtrate was lyophilized to obtain compound 1 as a white solid. The structure of compound 1 was verified by H NMR and mass spectrometry. HRMS (ESI) - : Theoretical value C 8 H 8 N 3 O 6 S - =274.0139, measured value: [M-Na] - =274.0147.

[0184] Example 2 Synthesis of Compound 2

[0185]

[0186] Step a: Synthesis of compound 2A:

[0187] Compound 1A (0.75 g, 1.51 mmol) was dissolved in tetrahydrofuran (10 mL) in a 25 mL single-necked bottle, and N-hydroxyphthalimide (300 mg, 1.81 mmol) and triphenylphosphine (480 mg, 1.81 mmol) were added at room temperature. The reaction bottle was placed in an ice bath at 0°C, and diisopropyl azodicarboxylate (370 mg, 1.81 mmol) was added to the reaction solution under stirring. The reaction solution was yellow, clear and transparent, and stirred at room temperature for 6 hours. The reaction was monitored by TLC until it was complete. The solvent was evaporated under reduced pressure to obtain a yellow oily crude product, which was purified by column chromatography to obtain 870 mg of a yellow oily product 2A with a yield of 89.7%.

[0188] LCMS: [M+H] + Theoretical value C 36 H 41 N 2 O 7 Si=641.3,measured value [M+H] + =641.5.

[0189] Step b: Synthesis of compound 2B:

[0190] Compound 2A (5 g, 7.8 mmol) was dissolved in methanol (75 mL) in a 250 mL single-necked bottle, cerium trichloride (1.92 g, 7.8 mmol) was added, and the mixture was stirred in an ice bath at 0°C for 15 minutes. Sodium borohydride (150 mg, 3.9 mmol) was slowly added to the reaction solution, and the reaction was stirred for 30 minutes. After TLC monitoring of the reaction was complete, a saturated ammonium chloride solution (50 mL) was added to the reaction solution to quench the reaction, and methyl tert-butyl ether (50 mL × 3) was added for extraction. The organic phases were combined, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain a yellow solid crude product, which was purified by chromatography to obtain 2.16 g of a yellow solid product 2B with a yield of 43.1%.

[0191] LCMS: [M+H] + Theoretical value C 36 H 43 N 2 O 7 Si=643.3,measured value:[M+H] + =643.3.

[0192] Step c: Synthesis of compound 2C:

[0193] Compound 2B (5.5 g, 8.56 mmol) was dissolved in dichloromethane (110 mL) in a 250 mL single-necked reaction bottle, and N-(benzyloxy)-2-nitrobenzenesulfonamide (3.96 g, 12.84 mmol), triphenylphosphine (2.92 g, 11.13 mmol) and diisopropyl azodicarboxylate (2.25 g, 11.13 mmol) were added in sequence at room temperature. The reaction solution was brown, clear and transparent, and stirred at room temperature overnight. After the reaction was completed under TLC monitoring, the solvent was evaporated under reduced pressure to obtain a yellow solid crude product, which was purified by chromatography to obtain 7.11 g of yellow solid product 2C, with a yield of 89.1%.

[0194] LCMS: [M+H] + Theoretical value C 49 H 53 N 4 O 11 SSi=933.3,measured value:[M+H] + =933.3.

[0195] Step d: Synthesis of compound 2D:

[0196] Compound 2C (3.83 g, 4.1 mmol) was dissolved in tetrahydrofuran (60 mL) in a 250 mL single-necked bottle, and acetic acid (0.37 g, 6.15 mmol) and tetrabutylammonium fluoride (6.15 mL, 6.15 mmol, 1 M in THF) were added at room temperature. The reaction solution was yellow, transparent and clear. It was stirred at room temperature for 6 hours. After the raw material was completely consumed as determined by LCMS, the solvent was evaporated under reduced pressure to obtain a yellow solid crude product, which was purified by chromatography to obtain 1.68 g of a yellow solid product 2D in a yield of 58.9%.

[0197] LCMS: [M+H] + Theoretical value C 33 H 35 N 4 O 11 S=695.2,measured value:[M+H] + =695.2.

[0198] Step e: Synthesis of compound 2E:

[0199] Periodic acid (7g, 30.71mmol) and chromium trioxide (28mg, 0.28mmol) were dissolved in a solution of acetonitrile (69.48mL) and water (0.52mL). Compound 2D (2.35g, 3.38mmol) was dissolved in a wet acetonitrile solution (60mL, acetonitrile 99.25%, water 0.75%) in a 250mL single-necked bottle, and the prepared oxidant solution was added using a constant pressure dropping funnel in an ice bath at 0°C. The reaction solution was white and turbid. The reaction was continued to stir at 0°C for 30 minutes. LCMS was used to determine whether the raw material was completely consumed. Then, sodium chloride aqueous solution (100mL) was added to the reaction solution and stirred, and the product was extracted with ethyl acetate (50mL×3). The organic phase was washed with saturated brine and the solvent was dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure to obtain 2.28g of a crude green solid product 2E with a yield of 95%, which was directly used in the next step.

[0200] LCMS: [M+H] + Theoretical value C 33 H 33 N 4 O 12 S=709.2,measured value:[M+H] + =709.1.

[0201] Step f: Synthesis of compound 2F:

[0202] Compound 2E (2.58 g, 3.64 mmol) was dissolved in N,N-dimethylformamide (50 mL) in a 250 mL single-necked bottle, and cesium carbonate (2.37 g, 7.28 mmol) and iodomethane (1.03 g, 7.28 mmol) were added at room temperature. The reaction solution was brown, clear and transparent. After stirring at room temperature for 15 minutes, the raw material was completely consumed as determined by LCMS. Then, sodium chloride aqueous solution was added to the reaction solution and stirred, and the product was extracted with ethyl acetate (50 mL×3). The organic phase was washed with saturated brine, the solvent was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain 1.31 g of yellow oily product 2F with a yield of 68.69%, which was directly used in the next step.

[0203] LCMS: [M+H] + Theoretical value C 34 H 35 N 4 O 12 S=723.2,measured value:[M+H] + =723.3.

[0204] Step g: Synthesis of compound 2G:

[0205] Compound 2F (2.67 g, 3.69 mmol) was dissolved in ethanol (50 mL) in a 100 mL single-necked bottle, and hydrazine monohydrate (0.74 g, 14.76 mmol) was added at room temperature. The reaction solution was yellow, clear and transparent. The reaction was stirred at room temperature for 2 hours. The raw material was completely consumed by monitoring by LCMS. Then, sodium chloride aqueous solution was added to the reaction solution and stirred, and the product was extracted with dichloromethane (50 mL × 3). The organic phase was washed with saturated brine and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, a yellow oily crude product was obtained, which was purified by chromatography to obtain 817 mg of a yellow oily product 2G with a yield of 37.32%.

[0206] LCMS: [M+H] + Theoretical value C 26 H 33 N 4 O 10 S=593.2,measured value:[M+H] + =593.1.

[0207] Step h: Synthesis of compound 2H:

[0208] Compound 2G (817 mg, 1.38 mmol) was dissolved in methanol (25 mL) in a 100 mL single-necked bottle, and sodium methoxide (0.11 g, 2.07 mmol) was added at room temperature. The reaction solution was yellow, clear and transparent. The reaction was stirred for 1.5 hours. The raw material was completely consumed by LCMS. Then, sodium chloride aqueous solution was added to the reaction solution and stirred, and the product was extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine, and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, a yellow solid crude product was obtained, which was purified by chromatography to obtain 365 mg of a yellow solid product 2H with a yield of 47.23%.

[0209] LCMS: [M+H] + Theoretical value C 25 H 29 N 4 O 9 S=561.2,measured value:[M+H] + =561.1.

[0210] Step i: Synthesis of compound 2I:

[0211] Compound 2H (365 mg, 0.65 mmol) was dissolved in dichloromethane (10 mL) in a 50 mL single-necked bottle, and zinc bromide (0.59 g, 2.6 mmol) was added at room temperature. Gas was generated during the reaction, and the mixture was stirred at room temperature overnight. The raw material was completely consumed as determined by LCMS. Ammonia water (20 mL) was then added to the reaction solution and stirred for 30 min, and the product was extracted with dichloromethane (20 mL × 3). The organic phase was washed with saturated brine, and the solvent was dried over anhydrous sodium sulfate. After the solvent was distilled off under reduced pressure, 275 mg of yellow solid product 2I was obtained, with a yield of 91.72%.

[0212] LCMS spectrum, [M+H] + Theoretical value C 20 H 21 N 4 O 7 S=461.1,measured value:[M+H] + =460.9.

[0213] Step j: Synthesis of compound 2J:

[0214] Compound 2I (275 mg, 0.6 mmol) was dissolved in methanol (5 mL) in a 25 mL single-necked reaction bottle, potassium carbonate (0.41 g, 3 mmol) and thioglycolic acid (0.14 g, 1.5 mmol) were added at room temperature, and the mixture was stirred overnight at room temperature. The reaction solution was yellow, clear and transparent, and the raw material was completely consumed as determined by LCMS. Then, sodium chloride aqueous solution (20 mL) was added to the reaction solution and stirred, and the product was extracted with dichloromethane (20 mL×3). The organic phase was washed with saturated brine, and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, a yellow solid crude product was obtained, which was purified by chromatography to obtain 75 mg of a yellow solid product 2J with a yield of 45.62%.

[0215] LCMS spectrum, [M+H] + Theoretical value C 14 H 18 N 3 O 3 =276.1, measured value: [M+H] + =276.1.

[0216] Step k: Synthesis of compound 2K:

[0217] Compound 2J (75 mg, 0.27 mmol) was dissolved in acetonitrile (3 mL) in a 10 mL three-necked bottle, and N,N-diisopropylethylamine (140 mg, 1.08 mmol) was added. After the reaction solution was cooled to -30°C, a solution of triphosgene (32 mg, 0.11 mmol) in acetonitrile (2 mL) was slowly added dropwise to the reaction solution, and the mixture was gradually returned to room temperature and stirred overnight. The raw material was completely consumed as determined by LCMS. The solvent was then evaporated under reduced pressure to obtain a yellow solid crude product, which was purified by chromatography to obtain 61 mg of a yellow solid product 2K with a yield of 74.39%.

[0218] LCMS: [M+H] + Theoretical value C 15 H 16 N 3 O 4 =302.1, measured value: [M+H] + =302.0.

[0219] Step 1: Synthesis of compound 2L:

[0220] Compound 2K (61 mg, 0.2 mmol) was dissolved in methanol (2 mL) in a 10 mL single-necked bottle, and 10% Pd / C (21.39 mg, 0.2 mmol) was added and stirred for 30 minutes under a hydrogen atmosphere. The raw material was completely consumed as determined by LCMS. The mixture was then filtered through a sand core filter padded with diatomaceous earth, and the filtrate was decompressed to distill off the solvent to obtain a yellow solid crude product, which was purified by chromatography to obtain 30 mg of a yellow solid product 2L, with a yield of 71.42%.

[0221] LCMS: [M+H] + Theoretical value C 8 H 10 N 3 O 4 =212.1, measured value: [M+H] + =212.2.

[0222] Step m: Synthesis of compound 2:

[0223] Compound 2L (30 mg, 0.14 mmol) was dissolved in pyridine (2 mL) in a 10 mL single-necked reaction bottle, pyridine / sulfur trioxide (133.65 mg, 0.84 mmol) was added, and the mixture was stirred at room temperature overnight. The raw material was completely consumed by LCMS. The solvent was then evaporated under reduced pressure to obtain a yellow solid crude product, which was purified by chromatography column. The product was dissolved in 5 mL of purified water and passed through a sodium ion exchange resin, washed with purified water, and the filtrate was lyophilized to obtain 32 mg of a yellow solid product 2, with a yield of 73.58%. The structure of compound 2 was verified by nuclear magnetic resonance hydrogen spectrum and mass spectrometry. HRMS (ESI) -: theoretical value C8 H 8 N 3 O 7 S - =290.2265, measured value: [M-Na] - =290.2247.

[0224] Example 3 Synthesis of Compound 3

[0225]

[0226] Step a: Synthesis of compound 3A:

[0227] Compound M-15 (1 g, 1.64 mmol) was dissolved in MeOH (50 mL) in a 250 mL single-mouth reaction bottle. The reaction bottle was placed in an ice bath and CeCl was added. 3 (0.4 g, 1.64 mmol) and then slowly add NaBH 4 (0.82mmol, 0.03g), stirred for 30 minutes. After the reaction was complete as monitored by TLC, saturated aqueous ammonium chloride solution (25mL) and water (25mL) were added to the reaction solution, and the product was extracted with ethyl acetate (50mL×3). The organic phases were combined, the extract was washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a yellow oily crude product. After purification by chromatography, 1g of colorless oily product 3A was obtained, with a yield of 99.7%.

[0228] 1 H NMR (400 MHz, CDCl 3 )δ7.65-7.60(m,4H),7.44-7.35(m,6H),5.93(d,J=20.2Hz,1H),4.50(d,J=10.3Hz,1H),4.40-4.05(m,4H),3.94(m,1H), 3.72(t,J=11.3Hz,1H),3.05(dt,J=19.6,10.6Hz,1H),1.78(m,1H),1.43(s,9H),1.04(s,9H),0.86(s,9H),0.00(s,6H).

[0229] Step b: Synthesis of compound 3B:

[0230] Compound 3A (1 g, 1.63 mmol) was dissolved in toluene (10 mL) in a 100 mL single-necked reaction bottle. The reaction bottle was placed in an ice bath and PPh 3(0.51 g, 1.96 mmol), BnONH-Ns (0.60 g, 1.96 mmol) and DIAD (0.40 g, 1.96 mmol) were added and stirred at room temperature for 2 hours. After the reaction was completed under TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a brown oily crude product, which was purified by chromatography to obtain 1.05 g of colorless oily liquid 3B, with a yield of 71.2%.

[0231] 1 H NMR (400 MHz, CDCl 3 )δ8.07(d,J=7.9Hz,1H),7.78-7.70(m,1H),7.67-7.51(m,7H),7.38(m,7H),7.29(s,3H),4.90(d,J=12.0Hz,2H),4.76(s,1H),4.54(s,2H),4 .07-3.98(m,2H),3.87(d,J=42.0Hz,1H),3.73(d,J=10.7Hz,1H),3.62 (d,J=13.8Hz,1H),1.20(s,9H),1.01(s,9H),0.80(s,9H),0.00(s,6H).

[0232] Step c: Synthesis of compound 3C:

[0233] Compound 3B (1.05 g, 1.16 mmol) was dissolved in 15 mL AcOH / THF / H2O in a 100 mL single-necked reaction bottle. 2 O (volume ratio 3:1:1), stirred at 60°C for 8 hours. After the reaction was completed as monitored by TLC, the pH of the reaction solution was adjusted to 7 with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate (15 mL×3), the organic phases were combined and the extracts were washed with saturated brine, the organic phases were dried over anhydrous sodium sulfate after separation, filtered, and concentrated under reduced pressure to obtain a yellow oily crude product, which was purified by chromatography to obtain 0.6 g of colorless oily liquid 3C with a yield of 65.4%.

[0234] 1H NMR(400MHz,DMSO)δ8.06(m,1H),8.01(m,1H),7.88(q,J=8.0Hz,1H),7.60 -7.38(m,10H),7.31(m,6H),5.71(brs,1H),5.08(t,J=5.2Hz,1H),4.80(d, J=8.8Hz,1H),4.74(s,1H),4.58(s,1H),4.38(d,J=19.9Hz,1H),3.96(d,J= 4.0Hz, 2H), 3.81 (s, 2H), 3.44 (d, J = 19.9Hz, 1H), 1.17 (s, 9H), 0.96 (s, 9H).

[0235] LC-MS: 3C molecular weight C 41 H 49 N 3 O 9 SSi=787.30,measured value:[M+H] + =788.4.

[0236] Step d: Synthesis of compound 3D:

[0237] Compound 3C (0.60 g, 0.76 mmol) was dissolved in 10 mL of dichloromethane in a 100 mL single-mouth reaction bottle, and Dess-Martin oxidant (0.39 g, 0.91 mmol) was slowly added to the reaction solution at room temperature, and stirred at room temperature for 1 hour. After TLC monitoring of the reaction was complete, 10 mL of n-hexane was added to the reaction solution, followed by 40 mL of saturated sodium thiosulfate aqueous solution and stirred for 10 minutes. After separation, 40 mL of saturated sodium bicarbonate aqueous solution was added to the organic phase and stirred for 10 minutes. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 0.6 g of a pale yellow oily crude product with a yield of 99.9%. The crude product was directly used in the next step without further purification.

[0238] 1 H NMR (400 MHz, CDCl 3 )δ8.95(s,1H),8.03(d,J=7.4Hz,1H),7.75(t,J=7.3Hz,1H),7.68-7.38(m,6H),7.37-7.02(m,9H),7.11(d,J=2.6Hz,2H), 6.45-6.15(m,1H),4.86(s,2H),4.71(d,J=9.9Hz,1H),4.59(d,J=4.7Hz,1H),3.90-3.31(m,4H),1.21(s,9H),0.91(s,9H).

[0239] LC-MS: 3D molecular weight C 41 H 47 N 3 O 9 SSi=785.28,measured value:[M+H] + =786.3.

[0240] Step e: Synthesis of compound 3E:

[0241] Compound 3D (0.6 g, 0.76 mmol) was dissolved in 10 mL of toluene solution in a 50 mL single-necked reaction bottle, and then (All) was added. 2 N-NH 2 (0.17 g, 1.52 mmol) and TFA (8 mg, 0.076 mmol), heated to reflux for 0.5 h. After the reaction was completed as monitored by TLC, it was concentrated under reduced pressure to obtain a pale yellow oily crude product. After purification by chromatography column, 0.22 g of colorless oily liquid 3E was obtained. The yield was 33.0%.

[0242] 1 H NMR(400MHz,DMSO)δ8.19-8.02(m,3H),7.98-7.90(m,1H),7.67-7.55(m,4H),7.51 -7.29(m,9H),7.21(t,J=8.2Hz,1H),6.92(s,1H),5.83-5.56(m,3H),5.11(dd,J=17 .3,8.8Hz,4H),5.24(s,1H),4.80(d,J=5.7Hz,1H),4.70(s,1H),4.57-4.45(m,2H) ,3.91-3.70(m,6H),3.54(d,J=10.8Hz,1H),3.12(s,1H),1.27(s,9H),0.99(s,9H).

[0243] LC-MS: 3E molecular weight C 47 H 57 N 5 O 8 SSi=879.37,measured value:[M+H] + =880.3.

[0244] Step f: Synthesis of compound 3F:

[0245] Compound 3E (220 mg, 0.25 mmol) was dissolved in 5 mL of dichloromethane solution in a 50 mL single-mouth reaction bottle, followed by the addition of zinc bromide (168.9 mg, 0.75 mmol), and stirred overnight at room temperature. After TLC monitoring of the reaction was complete, 20 mL of saturated sodium bicarbonate aqueous solution was added to the reaction solution, and after separation, the solution was washed with 20 mL of saturated sodium chloride aqueous solution, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 124.8 mg of a yellow oily crude product with a yield of 64.3%. The product was used directly in the next step without further purification.

[0246] 1 H NMR (400 MHz, CDCl 3 )δ8.04-7.98(m,1H),7.68(td,J=7.7,1.0Hz,1H),7.61-7.49(m,6H),7.36 -7.21(m,11H),6.67(s,1H),5.65-5.57(m,3H),5.23(s,2H),5.05-4.93(m ,4H),4.81(d,J=9.6Hz,1H),4.34(d,J=2.6Hz,1H),3.89-3.81(m,3H),3.6 1(d,J=5.4Hz,4H),3.17(dd,J=13.5,4.8Hz,1H),2.72(s,1H),0.95(s,9H).

[0247] LC-MS: 3F molecular weight C 42 H 49 N 5 O 6 SSi=779.32,measured value:[M+H] + =780.4.

[0248] Step g: Synthesis of compound 3G:

[0249] Compound 3F (124.8 mg, 0.16 mmol) was dissolved in 5 mL of methanol solution in a 25 mL single-mouth reaction bottle, followed by addition of potassium carbonate (110.6 mg, 0.8 mmol) and thioglycolic acid (36.8 mg, 0.4 mmol), and stirred at room temperature overnight. After TLC monitoring of the reaction was complete, aqueous solution (10 mL) was added to the reaction solution, extracted with ethyl acetate (10 mL×2), the organic phases were combined, washed with saturated brine (20 mL), and the organic phases were dried over anhydrous sodium sulfate after separation, filtered, and concentrated under reduced pressure to obtain a yellow oily crude product HZ7, which was separated by chromatography to obtain 71.4 mg of a light yellow oily liquid product 3G, with a yield of 75.2%.

[0250] 1H NMR (400 MHz, CDCl 3 )δ7.59(td,J=7.9,1.5Hz,4H),7.36-7.20(m,11H),6.66(s,1H),5.64-5.56(m,3H),5.03-4.93(m,4H),4.67(s,2H),3.97-3.91(m, 1H), 3.64 (t, J = 8.8Hz, 1H), 3.57 (d, J = 5.6Hz, 4H), 3.38 (s, 1H), 3.04 (d, J = 11.9Hz, 1H), 2.87 (dd, J = 12.2, 3.1Hz, 1H), 0.98 (s, 9H).

[0251] LC-MS: 3G molecular weight C 36 H 46 N 4 O 2 Si=594.34,measured value:[M+H] + =595.4.

[0252] Step h: Synthesis of compound 3H:

[0253] Compound 3G (71.4 mg, 0.12 mmol) and DIPEA (62.1 mg, 0.48 mmol) were dissolved in 62 mL of acetonitrile solution in a 100 mL single-mouth reaction bottle. The reaction bottle was placed in an ice bath. Triphosgene (14.2 mg, 0.048 mmol) was dissolved in 14 mL of acetonitrile and slowly added dropwise to the reaction solution. After the addition was complete, the mixture was stirred at room temperature overnight. After the reaction was completed by TLC monitoring, the reaction solution was directly concentrated and separated by column chromatography to obtain 62.1 mg of light yellow oily liquid product 3H, with a yield of 83.3%.

[0254] 1 H NMR (400 MHz, CDCl 3 )δ7.61-7.46(m,4H),7.39-7.21(m,10H),7.17(dd,J=6.4,2.7Hz,1H),6.74(s,1H),5.76-5.56(m,3H),5.42(d,J=6.3Hz,1H),5.10-4.96(m,4 H),4.87-4.79(m,1H),4.73(s,2H),4.59(t,J=13.8Hz,1H),4.01-3.86(m,1H),3.77(dd,J=14.9,3.9Hz,1H),3.73-3.66(m,4H),0.95(s,9H).

[0255] LC-MS: 3H molecular weight C 37 H44 N 4 O 3 Si=620.32, measured value: [M+H] + =621.3.

[0256] Step i: Synthesis of compound 3I:

[0257] Compound 3H (62.1 mg, 0.1 mmol) was dissolved in 4 mL of tetrahydrofuran in a 25 mL single-mouth reaction bottle, and then TBAF (1 M in THF) (0.2 mL, 0.2 mmol) was added to the reaction solution, and stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the reaction solution was directly concentrated and separated by column chromatography to obtain 31.3 mg of light yellow oily liquid product 3I, with a yield of 82.4%.

[0258] 1 H NMR (400 MHz, CDCl 3 )δ7.38-7.33(m,2H),7.33-7.25(m,3H),6.64(s,1H),6.00(d,J=5.1Hz, 1H),5.73-5.63(m,2H),5.12-5.02(m,4H),4.93(d,J=11.6Hz,1H),4.79( d,J=11.6Hz,1H),4.31(dd,J=7.8,5.1Hz,1H),4.18-4.09(m,1H),3.72(d ,J=5.5Hz,4H),3.69-3.61(m,1H),3.47-3.40(m,1H),3.12-2.98(m,2H).

[0259] LC-M: 3I molecular weight C 21 H 26 N 4 O 3 =382.20, measured value: [M+H] + =383.1.

[0260] Step j: Synthesis of compound 3J:

[0261] Periodic acid (1 g, 5.21 mmol) and chromium trioxide (2 mg, 0.02 mmol) were dissolved in 20 mL of acetonitrile solution and 0.15 mL of water at room temperature. Compound 3I (31.3 mg, 0.082 mmol) was dissolved in 5 mL of acetonitrile solution in a 50 mL single-mouth reaction bottle, the reaction bottle was placed in an ice bath, the above-prepared oxidant solution (4 mL) was added to the reaction solution, and the reaction was stirred at room temperature for 30 minutes. The raw material was completely consumed by LC-MS, and the target molecular weight of HZ10 appeared. Subsequently, 20 mL of water and 20 mL of saturated brine were added to the reaction solution, extracted with ethyl acetate (10 mL × 3), the organic phases were combined and dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 27.7 mg of a yellow oily crude product with a yield of 85.3%. The product was directly used for the next step reaction.

[0262] LCMS: 3J molecular weight C 21 H 24 N 4 O 4 =396.18, measured value: [M-OBn] + =289.1.

[0263] Step k: Synthesis of compound 3K:

[0264] Compound 3J (27.7 mg, 0.07 mmol) was dissolved in 3 mL of dichloromethane in a 25 mL single-necked reaction bottle, and then Pd(PPh 3 ) 4 (16.18 mg, 0.014 mmol), AcOH (6.3 mg, 0.1 mmol) and 1,3-dimethylbarbituric acid (DMBA) (65.6 mg, 0.42 mmol) were purged three times and then replaced with N 2 The reaction mixture was stirred at 40°C for 3 hours, and LC-MS showed that the raw material was completely consumed and the target molecular weight of 3K appeared. Then 5 mL of water was added to the reaction solution, and the solution was separated and washed with dichloromethane (5 mL×2). The aqueous phase was freeze-dried to obtain 14.7 mg of yellow solid 3K, with a yield of 66.7%.

[0265] Step 1: Synthesis of compound 3L:

[0266] Compound 3K (14.7 mg, 0.046 mmol) was dissolved in 3 mL of N,N-dimethylformamide in a 25 mL single-mouth reaction bottle, followed by the addition of HATU (26.23 mg, 0.069 mmol) and DIPEA (23.78 mg, 0.184 mmol), and stirred at room temperature for 3 hours. LC-MS showed that the raw materials were completely consumed. 5 mL of ethyl acetate was then added to the reaction solution, and washed with saturated sodium bicarbonate aqueous solution (6 mL) and saturated brine (6 mL). After separation, the organic phase was dried, filtered, and concentrated under reduced pressure to obtain a yellow oily crude product. After separation by chromatography, 12.2 mg of yellow oily liquid product 3L was obtained, with a yield of 86.9%.

[0267] Step m: Synthesis of compound 3M:

[0268] In a 25 mL single-necked reaction bottle, compound 3L (12.2 mg, 0.04 mmol) was dissolved in 3 mL

[0269] 10% Pd / C (4.3 mg, 0.04 mmol) was added to the methanol solution and stirred for 30 minutes under a hydrogen atmosphere. After the reaction was completed by TLC monitoring, the mixture was filtered using a sand core funnel with diatomaceous earth pad, and the filtrate was concentrated under reduced pressure to obtain a yellow oily crude product. After separation by chromatography, 7.9 mg of a yellow oily liquid product 3M was obtained, with a yield of 95.2%.

[0270] Step n: Synthesis of compound 3:

[0271] In a 25 mL single-necked reaction bottle, compound 3M (7.9 mg, 0.038 mmol) was dissolved in 3 mL

[0272] Sulfur trioxide pyridine (36.29 mg, 0.228 mmol) was added to the pyridine solution and stirred overnight at room temperature. After the reaction was complete as monitored by LC-MS, the reaction solution was concentrated under reduced pressure to obtain a yellow oily crude product, which was separated by column chromatography. The product was dissolved in 5 mL of purified water and passed through a sodium ion exchange resin, washed with purified water, and the filtrate was freeze-dried to obtain 9.5 mg of a yellow solid product 3 with a yield of 86.5%. The structure of compound 3 was verified by nuclear magnetic resonance hydrogen spectrum and mass spectrometry. HRMS (ESI) - : Theoretical value C 8 H 7 N 4 O 6 S - =287.0092, measured value: [M-Na] - =287.0105.

[0273] Example 4 Synthesis of Compound 4

[0274]

[0275] Step a: Synthesis of compound 4A:

[0276] Compound M-15 (610 mg, 1.0 mmol) was dissolved in methanol (20 mL) in a 50 mL single-necked reaction bottle, cerium trichloride (270 mg, 1.1 mmol) was added, and sodium borohydride (42 mg, 1.1 mmol) was added to the reaction solution in batches after cooling and stirring in an ice-water bath for 10 min. Bubbles were generated, and the clear liquid became a suspension. The reaction was carried out at 0°C for about 30 minutes. After the reaction was completed by TLC monitoring, an aqueous solution of ammonium chloride (10 mL) was added to the reaction solution and stirred, and the product was extracted with ethyl acetate (10 mL × 3). The extract was washed with water and saturated brine, and anhydrous Na 2 SO 4 The solvent was dried and the solvent was distilled off under reduced pressure to obtain a colorless glassy solid crude product which was purified by column chromatography to obtain 480 mg of a colorless glassy solid product 4A with a yield of 78.43%.

[0277] 1 H NMR (400 MHz, CDCl 3 )δ=7.63(t,J=8.0Hz,4H),7.37-7.42(m,6H),5.93(d,J=20.8Hz,1H),4.50(d,J=10.2Hz,1H),4.09-4.37(m,5H),3.93(dd,J 1 =41.6Hz,J 2 =10.0Hz,1H),3.70-3.73(m,1H),3.04(dt,J 1 =35.6Hz,J 2 =10.2Hz,1H),1.43(s,9H),1.04(s,9H),0.86(s,9H),0.00(d,J=9.6Hz,6H)ppm.

[0278] LCMS [M+H] + Theoretical value C 34 H 54 NO 5 Si 2 =612.35, measured value: [M+H] + =612.3.

[0279] Step b: Synthesis of compound 4B:

[0280] Compound 4A (1 g, 1.63 mmol) was dissolved in toluene (10 mL) in a 100 mL single-mouth reaction bottle, and triphenylphosphine (0.51 g, 1.96 mmol), BnONH-Ns (0.60 g, 1.96 mmol) and diisopropyl azodicarboxylate (0.40 g, 1.96 mmol) were added to the reaction bottle in an ice bath, and stirred at room temperature for 2 hours. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to obtain a brown oily crude product, which was purified by chromatography to obtain 1.05 g of colorless oily liquid 4B, with a yield of 71.2%.

[0281] 1 H NMR (400 MHz, CDCl 3 )δ=8.07(d,J=7.9Hz,1H),7.78-7.70(m,1H),7.67-7.51(m,7H),7.38(m ,7H),7.29(s,3H),4.90(d,J=12.0Hz,2H),4.76(s,1H),4.54(s,2H),4. 07-3.98(m,2H),3.87(d,J=42.0Hz,1H),3.73(d,J=10.7Hz,1H),3.62(d ,J=13.8Hz,1H),1.20(s,9H),1.01(s,9H),0.80(s,9H),0.00(s,6H)ppm.

[0282] Step c: Synthesis of compound 4C:

[0283] Compound 4B (1.05 g, 1.16 mmol) was dissolved in 15 mL AcOH / THF / H2O in a 100 mL single-necked reaction bottle. 2 O (volume ratio 3:1:1), stirred at 60°C for 8 hours. After the reaction was completed as monitored by TLC, the pH of the reaction solution was adjusted to 7 with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate (15 mL×3), the organic phases were combined and the extract was washed with saturated brine, the organic phase was dried over anhydrous sodium sulfate after separation, filtered, and concentrated under reduced pressure to obtain a yellow oily crude product, which was purified by chromatography to obtain 0.6 g of colorless oily liquid 4C with a yield of 65.4%.

[0284] 1H NMR (400MHz, DMSO) δ = 8.06 (m, 1H), 8.01 (m, 1H), 7.88 (q, J = 8.0Hz, 1H), 7.60 -7.38(m,10H),7.31(m,6H),5.71(brs,1H),5.08(t,J=5.2Hz,1H),4.80(d,J =8.8Hz,1H),4.74(s,1H),4.58(s,1H),4.38(d,J=19.9Hz,1H),3.96(d,J=4. 0Hz,2H),3.81(s,2H),3.44(d,J=19.9Hz,1H),1.17(s,9H),0.96(s,9H)ppm.

[0285] LC-MS: Z3 molecular weight C 41 H 49 N 3 O 9 SSi=787.30,measured value:[M+H] + =788.4.

[0286] Step d: Synthesis of compound 4D:

[0287] Prepare about 50 mL of wet acetonitrile solution (containing 0.75% water, volume ratio), dissolve periodic acid (570 mg, 2.52 mmol) and chromium trioxide (3 mg, 0.03 mmol) in 8 mL of wet acetonitrile to make them completely dissolved. Dissolve compound 4C (500 mg, 0.63 mmol) in wet acetonitrile (25 mL) in a 50 mL single-necked reaction bottle, gradually add the above periodic acid solution (6 mL, 1.89 mmol) dropwise under an ice-water bath, and the reaction system becomes turbid and orange. After about half an hour of reaction, add the above periodic acid solution (2 mL, 0.63 mmol) again, and continue to react for half an hour. After LCMS detection of the reaction, extract the product with ethyl acetate (10 mL×3), wash the extract with a 1:1 solution of water and saturated brine, and dry the solvent with anhydrous sodium sulfate. After the solvent is evaporated under reduced pressure, a pale yellow solid crude product 4D is obtained. The crude product is not purified and directly reacted in the next step.

[0288] LCMS: [M+H] + Theoretical value C 41 H 48 N 3 O 10 SSi=802.28,measured value:[M+H] + =802.2.

[0289] Step e: Synthesis of compound 4E:

[0290] In a 50 mL single-necked reaction bottle, the crude product of compound 4D (508 mg, 0.63 mmol), HATU (480 mg, 1.26 mmol), and ammonium chloride (100 mg, 1.89 mmol) were dissolved in DMF (10 mL). Diisopropylethylamine (0.42 mL, 2.52 mmol) was added dropwise at room temperature. The reaction system changed from a light yellow solution to a brown solution. The reaction was carried out for about two hours. The reaction was detected by LCMS. The product was extracted with ethyl acetate (10 mL × 3). The extract was washed with saturated sodium bicarbonate and saturated saline solution. Anhydrous Na 2 SO 4 The solvent was dried and the solvent was distilled off under reduced pressure to obtain a pale yellow solid crude product 4E, which was purified by chromatography to obtain 135.3 mg of a white solid Z5 with a yield of 26.8%.

[0291] 1 H NMR (400 MHz, CDCl 3 )δ=8.06(d,J=8.0Hz,1H),7.77(t,J=7.6Hz,1H),7.53-7.65(m,7H),7.30-7.41(m,11H),7.21-7.22(m,1H),6.33(brs,1H),5.18(s,1H), 4.85-4.91(m,2H),4.75(d,J=8.8Hz,1H),4.60(d,J=40.8Hz,1H),3.83-3.89(m,2H),3.51(d,J=8.8Hz,1H),1.20(s,9H),1.01(s,9H)ppm.

[0292] LCMS: [M+H] + Theoretical value C 41 H 49 N 4 O 9 SSi=801.30,measured value:[M+H] + =801.2.

[0293] Step f: Synthesis of compound 4F:

[0294] Compound 4E (96.9 mg, 0.12 mmol) and zinc bromide (140 mg, 0.6 mmol) were dissolved in dichloromethane (10 mL) in a 50 mL single-necked reaction bottle. Zinc bromide was dissolved and reacted at room temperature overnight. The product was extracted with dichloromethane (10 mL × 3). The extract was washed with saturated sodium bicarbonate and saturated saline solution. Anhydrous Na 2 SO 4 The solvent was dried and the solvent was distilled off under reduced pressure to obtain a pale yellow solid crude product Z6, which was purified by column chromatography to obtain 78 mg of a white solid 4F with a yield of 92.0%.

[0295] LCMS: [M+H] + Theoretical value C 36 H 41 N 4 O 7 SSi=701.24,measured value:[M+H] + =701.1.

[0296] Step g: Synthesis of compound 4G:

[0297] Compound 4F (83.9 mg, 0.12 mmol) and potassium carbonate (83.0 mg, 0.6 mmol) were dissolved in methanol (10 mL) in a 50 mL single-necked reaction bottle, and thioglycolic acid (28 mg, 0.3 mmol) was added dropwise under stirring at room temperature, and the reaction was allowed to proceed overnight at room temperature. After the reaction system was detected by TLC, the product was extracted with dichloromethane (10 mL × 3), and the extract was washed with saturated sodium bicarbonate and saturated saline solution, and the solvent was dried over anhydrous sodium sulfate. After the solvent was distilled off under reduced pressure, a pale yellow solid crude product Z7 was obtained, which was purified by chromatography to obtain 46 mg of a white solid 4G, with a yield of 74.5%.

[0298] LCMS: [M+H] + Theoretical value C 30 H 38 N 3 O 3 Si=516.27, measured value: [M+H] + =516.2.

[0299] Step h: Synthesis of compound 4H:

[0300] Compound 4G (103 mg, 0.2 mmol) and diisopropylethylamine (0.14 mL, 0.8 mmol) were dissolved in acetonitrile (10 mL) in a 50 mL single-necked reaction bottle and stirred at 0°C. Triphosgene (23.8 mg, 0.08 mmol) was dissolved in acetonitrile (2 mL) and added dropwise to the Z7 solution at a rate of 0.1 mL / min. After the addition was completed, the reaction was allowed to proceed overnight at room temperature. After the reaction was completed by TLC spot plate detection, the product was extracted with ethyl acetate (10 mL × 3), and the extract was washed with saturated saline solution and water, and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, a pale yellow solid crude product 4H was obtained, which was purified by column chromatography to obtain 88.3 mg of white solid Z7, with a yield of 81.5%.

[0301] LCMS: [M+H] + Theoretical value C 31 H 36 N 3 O 4Si=542.25, measured value: [M+H] + =542.3.

[0302] Step i: Synthesis of compound 4I:

[0303] Preparation of sodium hypobromite solution: Bromine (0.25 mL, 4.88 mmol), sodium hydroxide (1.9 g, 47.51 mmol) and water (100 mL) were mixed. 6.5 mL of sodium hypobromite solution was placed in a 50 mL single-necked reaction bottle, cooled to 0°C, and compound 4H (108 mg, 0.2 mmol) was added to the solution. The reaction system was heated to 65°C and reacted for 2 hours. After the reaction was completed by TLC detection, the product was extracted with ethyl acetate (10 mL × 3), and the extract was washed with sodium sulfite solution, saturated saline solution and water in turn, and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, a pale yellow solid crude product 4I was obtained, which was purified by column chromatography to obtain 41.1 mg of white solid 4I, with a yield of 40.1%.

[0304] LCMS: [M+H] + Theoretical value C 30 H 36 N 3 O 3 Si=514.25, measured value: [M+H] + =514.2.

[0305] Step j: Synthesis of compound 4J:

[0306] Compound 4I (103 mg, 0.2 mmol) and potassium carbonate (55.3 mg, 0.4 mmol) were dissolved in dichloromethane (10 mL) in a 50 mL single-necked reaction bottle and stirred at room temperature. Iodomethane (24.9 μL, 0.4 mmol) was added dropwise to the solution of 4I. After the addition was complete, the reaction was allowed to proceed overnight at room temperature. After the reaction was completed by monitoring by TLC, the product was extracted with ethyl acetate (10 mL × 3), and the extract was washed with saturated saline solution and water, and the solvent was dried over anhydrous sodium sulfate. After the solvent was distilled off under reduced pressure, a pale yellow solid crude product 4J was obtained, which was purified by column chromatography to obtain 86.5 mg of white solid 4J, with a yield of 82.0%.

[0307] LCMS: [M+H] + Theoretical value C 31 H 38 N 3 O 3 Si=528.27, measured value: [M+H] + =528.2.

[0308] Step k: Synthesis of compound 4K:

[0309] Compound 4J (106 mg, 0.2 mmol), sodium carbonate (63.6 mg, 0.6 mmol), and BocNH-OTs (114.9 mg, 0.4 mmol) were dissolved in 1,4-dioxane (10 mL) in a 50 mL single-necked reaction bottle and stirred at room temperature overnight. The reaction was monitored by TLC until completion, and the product was extracted with ethyl acetate (10 mL × 3), and the extract was washed with saturated saline solution and water, and anhydrous Na 2 SO 4 The solvent was dried and the solvent was distilled off under reduced pressure to obtain a pale yellow solid crude product 4K, which was purified by column chromatography to obtain 96.4 mg of a white solid 4K with a yield of 74.8%.

[0310] LCMS: [M+H] + Theoretical value C 36 H 47 N 4 O 5 Si=643.33, measured value: [M+H] + =643.3.

[0311] Step 1: Synthesis of compound 4L:

[0312] At 0 degrees Celsius, compound 4K (129 mg, 0.2 mmol) and tetrabutylammonium fluoride (261.5 mg, 1.0 mmol) were dissolved in tetrahydrofuran (5 mL) in a 50 mL single-necked reaction bottle, and stirred at 0 degrees Celsius for 3 hours. After the reaction was completed by TLC monitoring, the product was extracted with ethyl acetate (10 mL × 3), and the extract was washed with saturated saline solution and water, and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, a pale yellow solid crude product 4L was obtained, which was purified by column chromatography to obtain 75.2 mg of a white solid 4L, with a yield of 92.8%.

[0313] LCMS: [M+H] + Theoretical value C 20 H 29 N 4 O 5 =405.21, measured value: [M+H] + =405.2.

[0314] Step m: Synthesis of compound 4M:

[0315] Prepare about 50 mL of wet acetonitrile solution (containing 0.75% water, volume ratio), dissolve periodic acid (570 mg, 2.52 mmol) and chromium trioxide (3 mg, 0.03 mmol) in 8 mL of wet acetonitrile to make them completely dissolved. Dissolve compound 4L (255 mg, 0.63 mmol) in wet acetonitrile (25 mL) in a 50 mL single-necked reaction bottle, gradually add the above periodic acid solution (6 mL, 1.89 mmol) dropwise under an ice-water bath, and the reaction system becomes turbid and orange. After about half an hour of reaction, LCMS detects whether the reaction is complete. If the reaction is not complete, add the above periodic acid solution (2 mL, 0.63 mmol), continue to react for half an hour, extract the product with ethyl acetate (10 mL×3), wash the extract with a 1:1 solution of water and saturated brine, and dry the solvent with anhydrous sodium sulfate. After the solvent is evaporated under reduced pressure, a pale yellow solid crude product 4M is obtained, which is directly used in the next step reaction.

[0316] LCMS: [M+H] + Theoretical value C 20 H 27 N 4 O 6 =419.19, measured value: [M+H] + =419.1.

[0317] Step n: Synthesis of compound 4N:

[0318] Compound 4M (83.7 mg, 0.2 mmol) was dissolved in dichloromethane (10 mL) in a 50 mL single-necked reaction bottle. Trifluoroacetic acid (23 μL, 0.3 mmol) was added dropwise under stirring at room temperature and reacted overnight. The reaction system remained colorless and transparent. The pH value was adjusted to neutral, and the product was extracted with ethyl acetate (10 mL × 3). The extract was washed with saturated saline solution and water, and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, 43.9 mg of a pale yellow solid crude product 4N was obtained, with a yield of 68.8%.

[0319] LCMS: [M+H] + Theoretical value C 15 H 19 N 4 O 4 =319.14, measured value: [M+H] + =319.1.

[0320] Step o: Synthesis of compound 4O:

[0321] In a 50 mL single-necked reaction bottle, the crude product of compound 4N (200.5 mg, 0.63 mmol) and HATU (480 mg, 1.26 mmol) were dissolved in DMF (5 mL), and diisopropylethylamine (0.42 mL, 2.52 mmol) was added dropwise at room temperature. The reaction system changed from a light yellow solution to a brown solution. The reaction was monitored by TLC. The reaction lasted for about two hours. The product was extracted with ethyl acetate (10 mL × 3), and the extract was washed with saturated sodium bicarbonate and saturated saline solution, and the solvent was dried over anhydrous sodium sulfate. After the solvent was evaporated under reduced pressure, a light yellow solid crude product 4O was obtained, which was purified by column chromatography to obtain 83.2 mg of a white solid 4O, with a yield of 45.1%.

[0322] LCMS: [M+H] + Theoretical value C 15 H 16 N 4 O 3 =300.12, measured value: [M+H] + =300.0.

[0323] Step p: Synthesis of compound 4:

[0324] In a 25 mL single-necked reaction bottle, compound 4O (30 mg, 0.1 mmol) was dissolved in 3 mL

[0325] 10% Pd / C (10.8 mg, 0.1 mmol) was added to the methanol solution and stirred for 30 min under a hydrogen atmosphere. After the reaction was complete as monitored by TLC, the mixture was filtered using a sand core funnel pad with diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain a yellow oily crude product. The crude product was dissolved in 3 mL of pyridine solution, and sulfur trioxide pyridine (90.7 mg, 0.57 mmol) was added and stirred at room temperature overnight. After the reaction was complete as monitored by LCMS, the reaction solution was concentrated under reduced pressure to obtain a yellow oily crude product, which was separated by column chromatography. The product was dissolved in 5 mL of purified water and then passed through a sodium ion exchange resin, washed with purified water, and the filtrate was freeze-dried to obtain 18.9 mg of a yellow solid product 4 with a yield of 65.1%. The structure of compound 4 was verified by hydrogen nuclear magnetic resonance and mass spectrometry. HRMS (ESI) -: theoretical value C 8 H 9 N 4 O 6 S - =289.0248, measured value: [M-Na] - =289.0261.

[0326] Example 5 Synthesis of Compound 5

[0327]

[0328] Step a: Preparation of intermediate 5B

[0329] Tetrabutylammonium fluoride (1M in THF) (23.6mL, 23.6mmol) was added to a solution of intermediate 5A (4.58g, 5.9mmol) in tetrahydrofuran (100mL) at room temperature, and the color of the reaction solution became darker. The reaction was allowed to proceed overnight at room temperature, and TLC monitoring showed that the raw material had reacted completely. The mixture was extracted with ethyl acetate (50mL) and water (50mL), and after separation of the organic phase, the aqueous phase was extracted twice with ethyl acetate (30mL×2), the organic phases were combined, and the organic phases were concentrated onto silica gel. Silica gel column chromatography (PE / EA=0-70%) gave yellow oily intermediate 5B (2.4g, 62%).

[0330] LC-MS: [M] = C 31 H 42 N 4 O 10 S=662.26,[M+1]=663.3:

[0331] Step b: Preparation of intermediate 5C

[0332] Chromic anhydride (95mg, 0.95mmol) and periodic acid (9.5g, 41.6mmol) were added to a mixed solvent of acetonitrile (143ml) and water (1.5ml) and stirred at ambient temperature for 30 minutes. Intermediate 5B (9g, 13.6mmol) was dissolved in a mixed solvent of acetonitrile (40ml) and water (1ml), and the prepared oxidant was added dropwise under an ice-water bath. The process took 2 hours. After the addition was complete, the ice-water bath was removed and stirred at ambient temperature for 30 minutes. LC-MS showed that the raw material was completely consumed. Saturated brine (300ml) was added to quench the reaction, and DCM was extracted (200ml×2). The organic phase was dried and concentrated. No further purification was required and the next step was directly carried out.

[0333] The crude product obtained above was dissolved in DCM (40 ml) solution. Under ice-water bath, trifluoroacetic acid (20 ml) was added along the wall and stirred at 25°C for 1 hour. LC-MS showed that the raw material was completely consumed. Under ice-water bath, saturated sodium bicarbonate aqueous solution (violent bubbling) was slowly added until no more bubbles were generated. DCM extraction (100 ml × 2) was performed, the organic phase was dried, filtered and concentrated, and purified by column chromatography (DCM / MeOH = 30 / 1-10 / 1) to obtain the target compound 5C (4.9 g, 78.5%) as a light black solid.

[0334] LCMS:[M]:C 21 H 22 N 4 O 6S=458.13,MS(ESI,positive)found:[M+H] + =459.1

[0335] Step c: Preparation of intermediate 5D

[0336] Intermediate 5C (4.0 g, 8.72 mmol) was dissolved in acetonitrile (20 ml), p-toluene thiophenol (5.6 g, 45.0 mmol) and potassium carbonate (6.2 g, 45.0 mmol) were added at ambient temperature, and stirred overnight at room temperature. LC showed that the starting material was completely consumed. Filter, wash the filter cake with a mixed solvent of DCM / MeOH (10 / 1), and concentrate the filtrate. The product spot was eluted by column chromatography (MeCN / DCM=1 / 1-DCM / MeOH=10 / 1), and concentrated at 25°C to obtain a light black solid 5D (1.64 g, 68.6%).

[0337] LCMS:[M]:C 15 H 19 N 3 O 2 =273.15,MS(ESI,positive)found:[M+H] + =274.2

[0338] Step d: Preparation of intermediate 5E

[0339] Intermediate 5D (1.53 g, 5.6 mmol) and DIPEA (3.3 ml, 20.0 mmol) were dissolved in acetonitrile (500 ml), and a solution of triphosgene (0.59 g, 2.0 mmol) in acetonitrile (50 ml) was added dropwise under an ice-salt bath. The process took 4 hours. After the addition was completed, the mixture was stirred at 25°C for 1 hour. LC-MS showed that the reaction was complete. The mixture was concentrated at 35°C to remove acetonitrile, washed with dilute hydrochloric acid (0.5N, 150 ml), extracted with DCM (100 ml×2), and the organic phase was dried, filtered and concentrated (25°C). The mixture was purified by column chromatography (DCM / i-PrOH=40 / 1-10 / 1) to obtain a light yellow solid 5E (0.94 g, 55.9%).

[0340] LCMS:[M]:C 16 H 17 N 3 O 3 =299.13,MS(ESI,positive)found:[M+H] + =300.1.

[0341] Step e: Preparation of intermediate 5F

[0342] Intermediate 5E (0.1g, 0.33mmol), triethylamine (0.15ml, 1.05mmol), sulfur trioxide triethylamine complex (0.15g, 1.05mmol) and palladium carbon (5wt%, 70mg) were suspended in a mixed solvent of isopropanol (1ml), ethyl acetate (2ml) and water (3ml), replaced with a double-layer hydrogen balloon twice, stirred at ambient temperature for 1 hour, LC-MS showed that the starting material was completely consumed, the hydrogen balloon was removed, and stirring was continued at ambient temperature for 1 hour. Filter, the filter cake was washed with water, isopropanol / water = 1 / 1, and water in turn. The filtrate was washed with isopropyl acetate (20ml) and extracted with water (15ml×2). Tetrabutylammonium hydrogen sulfate (1.0g, 3.0mmol) was added to the aqueous phase and stirred at ambient temperature for 20 minutes. The mixture was washed with water (20 ml), extracted with DCM (15 ml×5), and the organic phase was concentrated (25° C.) and subjected to prep-HPLC to obtain the target compound 5F (46 mg, 25.8%).

[0343] LCMS: [M - ]:C 9 H 10 N 3 O 6 S - =288.03,MS(ESI,negative)found:[M] - =288.2

[0344] Step f: Preparation of compound 5

[0345] Dowex(R)50WX8-200-400 ion exchange resin (10.0 g) was adjusted by stirring with aqueous sodium hydroxide solution (0.2 N, 20 ml) for 30 minutes, filtered, and the filter cake was washed with water, acetone / water (1 / 1), and water in turn. Intermediate 21 (46 mg, 0.086 mmol) was dissolved in a mixed solvent of acetonitrile (5 ml) and water (5 ml), added with the above ion exchange resin, stirred at ambient temperature for 1.5 hours, filtered, and the filter cake was washed with water. The filtrate was lyophilized to obtain a light yellow compound 5 (11 mg, 40.7%). The structure of compound 5 was verified by nuclear magnetic resonance hydrogen spectrum and mass spectrometry. HRMS (ESI) -: theoretical value C 9 H 10 N 3 O 6 S - =288.0296, measured value: [M-Na] - =288.0308.

[0346] The technical solution of the present invention is further illustrated by experiments below.

[0347] Test Example 1 Antibacterial Activity Test

[0348] MIC is determined by the microdilution method according to the guidelines of The Clinical and laboratory Standard Institute (CLSI). The specific test process is as follows: Take out the agar plate that has been cultured for 18-24 hours to observe the morphology and size of the colonies, select several well-separated colonies, use a sterile inoculation loop to pick a part of the colonies for Gram staining microscopy, and directly make a bacterial suspension with the remaining part of the corresponding colonies in sterile saline, and adjust the concentration of the bacterial suspension to 0.5 McFarland units. Dilute the corrected bacterial solution with CAMHB broth to (4-8)×10 5 CFU / mL, ready for use. The bacterial solution was co-cultured with antibiotics and β-lactamase inhibitors in a 96-well plate at 37°C for 16-20 hours, and the lowest drug concentration that completely inhibited bacterial growth in the wells was the minimum inhibitory concentration.

[0349] Table 1 Antibacterial activity MIC 50 (mg / L)

[0350]

[0351]

[0352] Note: “+++” indicates MIC 50 ≤0.1mg / L; "++" means 0.1mg / L <MIC 50 ≤1mg / L; "+" means 1mg / L <MIC 50 ≤10mg / L; “-” indicates MIC 50 >10mg / L.

[0353] As can be seen from the above table, when the Example compounds are used in combination with the β-lactam antibiotic ceftazidime, the MIC of ceftazidime against several β-lactamase-producing resistant bacteria can be reduced several times, indicating that the Example compounds can effectively enhance the sensitivity of β-lactamase-producing resistant bacteria to β-lactam antibiotics.

[0354] Test Example 2 β-lactamase inhibitory activity test

[0355] The IC50 test of β-lactamase inhibitors against β-lactamase uses nitrocefin as a substrate. The enzyme is incubated with a gradient dilution of the β-lactamase inhibitor in 50 mM phosphate buffer (containing 0.1 mg / mL bovine serum albumin) at 37°C for 10 minutes. 100 μM nitrocefin is added to initiate the reaction, and the reaction temperature is set to 37°C. The absorbance value at 490 nm is recorded every minute for a total of 1 hour. The initial reaction rate is calculated and the IC is calculated in GraphPadPrsim software 50.

[0356] Table 2 Inhibitory activity IC of compounds against β-lactamase 50 (mg / L)

[0357]

[0358] Note: “+++” means IC 50 ≤1μM; "++" means 1μM <IC 50 ≤10μM; “+” means 10μM <IC 50 ≤100μM; “-” indicates MIC 50 >100μM.

[0359] As can be seen from the above table, the example compounds can exhibit good inhibitory activity against the selected three categories A / C / D, a total of 5 β-lactamases, indicating that the compounds of the present invention can enhance the sensitivity of bacteria to existing β-lactam antibiotics by inhibiting β-lactamase.

[0360] Test Example 3 Antibacterial Activity Test of Clinically Isolated Strains

[0361] The specific test process of antibacterial activity test was the same as that of Test Example 1: the bacterial suspension of different clinical isolates was diluted with CAMHB broth to (4-8)×10 5 CFU / mL, ready for use. The bacterial solution was co-cultured with antibiotics and β-lactamase inhibitors in a 96-well plate at 37°C for 16-20 hours, and the lowest drug concentration that completely inhibited bacterial growth in the wells was the minimum inhibitory concentration.

[0362] Table 3 Antibacterial activity MIC 50 (mg / L)

[0363]

[0364] Note: “+++” indicates MIC 50 ≤0.1mg / L; "++" means 0.1mg / L <MIC 50 ≤1mg / L; "+" means 1mg / L <MIC 50 ≤10mg / L; “-” indicates MIC 50 >10mg / L.

[0365] As can be seen from the above table, when compound 5 is used in combination with the β-lactam antibiotic ceftazidime or meropenem, the sensitivity of clinical strains of Escherichia coli and Klebsiella pneumoniae to ceftazidime or meropenem can be greatly enhanced, indicating that the example compounds can effectively enhance the sensitivity of carbapenem-resistant bacteria to β-lactam antibiotics.

[0366] It can be seen from the above embodiments and experimental examples that the present invention provides a class of β-lactamase inhibitors, which, when used in combination with traditional β-lactam antibiotics, can reverse the problem of bacterial resistance to β-lactam antibiotics due to the expression of β-lactamase, and have good application prospects in the clinical treatment of bacterial infections.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The structural formula of the compound of formula I is as follows: in, A1 is selected from -CO-; A2 is selected from -NH-; A3 is selected from -NR4-, -O-, -N= or -CH2-; A4 is selected from -CR5R6-, -CR7= or a bond; R1 is selected from R2 is selected from hydrogen; R4 is selected from methyl; R5 and R6 are selected from hydrogen; R7 is selected from hydrogen; M is selected from hydrogen and metal ions.

2. The compound according to any one of claims 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The structural formula of the compound is:

3. A method for preparing the compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The steps include: Step 1, using compound MI as a raw material to carry out a cyclization reaction to obtain intermediate A or intermediate B; Step 2, using intermediate A or intermediate B as a raw material to carry out a cyclization reaction to obtain a product; Wherein, A1, A2, A3, A4, R1, R2, R5, R6 are as described in claim 1 or 2; X is selected from O or NH; PG1 and PG2 are independently selected from hydrogen or an amine protecting group; PG3 and PG4 are independently selected from hydrogen or a hydroxyl protecting group; PG5 is selected from hydrogen, a hydroxyl protecting group or an amine protecting group.

4. The preparation method according to claim 3, characterized in that: The preparation method of the compound MI comprises the following steps: It is prepared by using compound M-II as a raw material, undergoing olefin metathesis reaction, ring closure, and then reduction and Mitsunobu reaction; Among them, R a1 and R a1 are independently selected from hydrogen or methyl.

5. Use of the compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a medicament for treating infection by drug-resistant bacteria expressing β-lactamase.

6. A drug for treating bacterial infection, characterized in that: The invention relates to a preparation which uses the compound described in claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as an active ingredient and adds pharmaceutically acceptable adjuvants or auxiliary ingredients.

7. The drug according to claim 6, characterized in that: The drug is used to treat infections caused by drug-resistant bacteria expressing β-lactamase.

8. A combined drug for treating bacterial infection, characterized in that: It comprises the drug according to claim 6 or 7 and at least one β-lactam antibiotic.

9. A compound preparation for treating bacterial infection, characterized in that: The invention relates to a compound preparation which is prepared by using the combined drug as claimed in claim 8 as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

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