HSP90 inhibitor and its preparation method and use

Abstract

The present invention discloses an HSP90 inhibitor, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, and a preparation method and use thereof. The compound of the general formula (I) provided by the present invention can be used as an HSP90 inhibitor, which can be used to prevent and / or treat HSP90-related diseases, such as tumors, cancers, organ fibrosis, and can also be used for antibacterial purposes. All compounds of the general formula (I) involved in the present invention are HSP90 inhibitors, having stronger HSP90 inhibitory activity, especially anti-tumor, anti-cancer, anti-organ fibrosis and antibacterial activity.

Description

Technical Field

[0001] The present invention relates to the field of drug synthesis, and in particular to an HSP90 inhibitor or a pharmaceutically acceptable salt thereof, and a preparation method and use thereof. Background Art

[0002] HSP90 is an important target for treating a variety of human diseases, including anti-tumor, anti-organ fibrosis, antibacterial, etc. Currently, no HSP90 inhibitor has been marketed for the treatment of diseases, so there is still an urgent need to develop new HSP90 inhibitors. Summary of the invention

[0003] Based on the needs of the prior art, the present invention provides an HSP90 inhibitor or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising the same, and a preparation method and use thereof.

[0004] The first aspect of the present invention provides a compound represented by general formula (I) or a pharmaceutically acceptable salt thereof:

[0005]

[0006] Wherein: R1 is selected from the group consisting of alkyl, aryl, methylene aryl and halogen; preferably, the halogen is a fluorine atom or a chlorine atom;

[0007] R2 is selected from the group consisting of para- or meta-O-Ph-CONHOH, para- or meta-O-CH2-Ph-CONHOH, para- or meta-O-Ph-CH=CH-CONHOH, para- or meta-CONHOH, para- or meta-CH=CH-CONHOH, para- or meta-SOCH3, para- or meta-SO2CH3, para- or meta-PO(CH3)2, and para- or meta-NO2;

[0008] L is absent of any substituent or is selected from the group consisting of alkyl, vinyl, and ethynyl; wherein the alkyl group is preferably -(CH2) n -, most preferably -(CH2) 1-2 -;

[0009] X is O or NH.

[0010] Preferably, the R1 is selected from C 1-6 Alkyl, C 6-10 Aryl, methylene (C 6-10 A group consisting of an aromatic group, a fluorine atom, and a chlorine atom.

[0011] Preferably, R2 is selected from the group consisting of para- or meta-O-Ph-CONHOH, para- or meta-O-CH2-Ph-CONHOH, para- or meta-O-Ph-CH=CH-CONHOH, para- or meta-CONHOH, para- or meta-CH=CH-CONHOH, para- or meta-SOCH3, para- or meta-SO2CH3, para- or meta-PO(CH3)2 and para- or meta-NO2.

[0012] Preferably, X is O or NH.

[0013] Preferably, L does not have any substituent or L is selected from the group consisting of methylene, ethylene, vinyl and ethynyl.

[0014] In some embodiments of the present invention, the compound or pharmaceutically acceptable salt thereof is selected from Table 1.

[0015] The second aspect of the present invention provides a pharmaceutical composition comprising an effective amount of the above compound or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable excipient or a pharmaceutically acceptable carrier.

[0016] The third aspect of the present invention provides use of the above compound or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in the preparation of an HSP90 inhibitor.

[0017] The fourth aspect of the present invention provides the HSP90 inhibitor for use in preventing and / or treating tumors, cancers or organ fibrosis.

[0018] The fifth aspect of the present invention provides the use of the HSP90 inhibitor for antibacterial purposes.

[0019] The beneficial effects of the present invention include at least:

[0020] The HSP90 inhibitor or its pharmaceutically acceptable salt or pharmaceutical composition of the present invention can be used to prevent and / or treat HSP90-related diseases, such as tumors, cancers, organ fibrosis, and can also be used for antibacterial. All compounds of the general formula (I) involved in the present invention are HSP90 inhibitors, having stronger HSP90 inhibitory activity, especially anti-tumor, anti-cancer, anti-organ fibrosis and antibacterial activity.

[0021] The features and advantages of the present invention will be described in detail in the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIG. 1( a ) and FIG. 1( b ) show the inhibition rate results of 112 representative compounds of the present invention on HSP90 at 200 nM.

[0023] FIG. 2( a ) and FIG. 2( b ) show the proliferation inhibitory activity of 112 representative compounds of the present invention on A549 lung cancer cells and lung fibroblasts HLF-1.

[0024] Figure 3 The results showed that compound HH-69 has an inhibitory activity against tumors.

[0025] Figure 4 Compound HH-69 was shown to reverse lung fibrosis in mice. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical scheme and beneficial effects of the present invention clearer, the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. Examples of the embodiments are shown in the accompanying drawings. It should be understood that the specific embodiments described in the following embodiments of the present invention are only exemplary descriptions of the specific embodiments of the present invention, and are intended to be used to explain the present invention, but do not constitute a limitation of the present invention.

[0027] The endpoints of the scope disclosed in this article and any value are not limited to this accurate scope or value, and these scopes or values ​​should be understood as comprising values ​​close to these scopes or values.For numerical range, between the endpoint values ​​of each scope, between the endpoint values ​​of each scope and a separate point value, and between separate point values, one or more new numerical ranges can be combined with each other, and these numerical ranges should be considered as specifically disclosed in this article.In the description of the application, unless otherwise stated, the meaning of similar words such as "one / a kind", "multiple / multiple" is two / kinds or more than two / kinds.

[0028]

Terminology

[0029] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0030] As used herein, when used in reference to a specific recited value, the term "about" means that the value may vary by no more than 1% from the recited value.

[0031] As used herein, the term "comprising" or "including" may be open, semi-closed and closed. Selected from the group consisting of means at least one or one of the listed items.

[0032] As used herein, definitions of standard chemical terms (eg, groups) can be found in the reference literature in the art.

[0033] Unless otherwise indicated, conventional methods within the technical scope of the art are adopted, such as mass spectrometry, NMR, IR and UV / VIS spectroscopy and pharmacological methods. Unless specifically defined, the terms used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and drugs and medicinal chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, preparation and delivery, and in the treatment of patients. For example, the instructions for use of the kit can be utilized by the manufacturer, or the reaction and purification can be implemented according to a manner well known in the art or the description of the present invention. Generally, the above-mentioned techniques and methods can be implemented according to conventional methods well known in the art according to the description in a plurality of summaries and more specific documents cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable structural parts and compounds.

[0034] When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result when the formula is written from right to left.

[0035] The section headings used herein are only for the purpose of organizing the article and should not be interpreted as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals and papers, are incorporated herein by reference in their entirety.

[0036] Certain chemical groups defined herein are preceded by a shorthand notation to indicate the total number of carbon atoms present in the group. 1-6 Alkyl refers to an alkyl group as defined below having a total of 1 to 6 carbon atoms. The total number of carbon atoms in the shorthand notation does not include carbons that may be present in substituents of the group being described.

[0037] In addition to the foregoing, when used in the specification and claims of the present application, the following terms have the meanings indicated below unless otherwise specifically stated.

[0038] As used herein, the term "compound of the present invention" or "active ingredient of the present invention" is used interchangeably to refer to a stereoisomer, enantiomer, or pharmaceutically acceptable salt thereof of a compound of the general formula. The term also includes racemates, optical isomers, isotopic compounds (such as deuterated compounds) or prodrugs.

[0039] "Stereoisomers" refer to compounds composed of the same atoms, bonded by the same bonds, but having different three-dimensional structures. The present invention is intended to encompass various stereoisomers and mixtures thereof.

[0040] When the compounds of the present invention contain olefinic double bonds, and unless specified otherwise, it is intended that the compounds of the present invention include both E- and Z- geometric isomers.

[0041] "Tautomer" refers to an isomer formed when a proton is shifted from one atom of a molecule to another atom of the same molecule. All tautomeric forms of the compounds of the present invention are intended to be encompassed within the scope of the present invention.

[0042] The compounds of the invention or their pharmaceutically acceptable salts may contain one or more chiral carbon atoms and may therefore give rise to enantiomers, diastereomers and other stereoisomeric forms. Each chiral carbon atom may be defined as (R)- or (S)- based on stereochemistry. The present invention is intended to include all possible isomers, as well as racemates and optically pure forms thereof. The compounds of the invention may be prepared using racemates, diastereomers or enantiomers as starting materials or intermediates. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0043] Conventional techniques for the preparation / isolation of individual isomers include chiral synthesis from appropriate optically pure precursors, or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high performance liquid chromatography.

[0044] The present invention also includes isotopically labeled compounds that are equivalent to the original compounds disclosed herein. However, in practice, it is common for one or more atoms to be replaced by atoms having a different atomic mass or mass number than the original atoms. Examples of isotopes of the compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes. The compounds of the present invention, or enantiomers, diastereomers, isomers, or pharmaceutically acceptable salts or solvates, which contain isotopes or other isotopic atoms of the above compounds are all within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as radioactive isotopes, are useful in tissue distribution experiments of drugs and substrates. For example, tritium, i.e. 3 H and carbon 14, i.e. 14 C, their preparation and detection are relatively easy. It is the first choice among isotopes. In addition, heavier isotope substitutions such as deuterium, i.e. 2 H, due to its good metabolic stability, has advantages in certain therapies, such as increasing half-life or reducing dosage in vivo, and therefore, can be preferred in certain cases. Isotope-labeled compounds can be prepared by general methods by replacing readily available isotope-labeled reagents with non-isotope reagents and using the schemes disclosed in the examples. In the present application, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0045] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, and the like. Organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetates, trifluoroformates, trifluoroacetates, propionates, caproates, caprylates, decanoates, undecylenates, glycolates, gluconates, pyroglutamic acid, citrates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, isethionates, and the like. These salts can be prepared by methods known in the art.

[0046] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, and the like. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following salts: primary, secondary, and tertiary amines, substituted amines, including natural substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, and the like. These salts can be prepared by methods known in the art.

[0047] As described herein, the compounds of the present invention can be replaced with any number of substituents or functional groups to expand their scope. Generally, the term "substituted" appears before or after the term "optional", and the general formula including substituents in the formulation of the present invention refers to replacing hydrogen free radicals with designated structural substituents. When multiple positions in a specific structure are substituted by multiple specific substituents, the substituents can be the same or different at each position. The term "substituted" used herein includes all allowed organic compound substitutions. In a broad sense, allowed substituents include non-cyclic, cyclic, branched non-branched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds. In the present invention, heteroatom nitrogen can have hydrogen substituents or any allowed organic compounds described above to supplement its valence. In addition, the present invention is not intended to limit the allowed substitution of organic compounds in any way. The present invention believes that the combination of substituents and variable groups is very good in the treatment of diseases in the form of stable compounds. As used herein, the term "stable" refers to compounds that are stable and that maintain the structural integrity of the compound over a period of time sufficient to be detected, and preferably over a period of time sufficient to be effective, as used herein for the purposes described above.

[0048] The metabolites of the compounds and pharmaceutically acceptable salts thereof involved in the present application, as well as prodrugs that can be converted into the structures of the compounds and pharmaceutically acceptable salts involved in the present application and in vivo, are also included in the claims of the present application.

[0049] Pharmaceutical compositions and methods of administration

[0050] The pharmaceutical composition of the present invention is used to prevent and / or treat cancer, tumors, organ fibrosis, and antibacterial, etc. In the present application, "pharmaceutical composition" refers to a preparation of a compound of the present invention and a medium generally accepted in the art for delivering biologically active compounds to mammals (e.g., humans). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote administration of an organism, facilitate the absorption of the active ingredient, and then exert biological activity. The term "pharmaceutical" as used herein refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compound of the present invention, and is relatively non-toxic, that is, the substance can be applied to an individual without causing adverse biological reactions or interacting with any component contained in the composition in an adverse manner.

[0051] In this application, "pharmaceutical excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for human or livestock use.

[0052] The "cancer" or "tumor" of the present invention includes, but is not limited to, lymphoma, sarcoma, melanoma, lung cancer, liver cancer, breast cancer, ovarian cancer, prostate cancer, bladder cancer, pancreatic cancer, thyroid cancer, laryngeal cancer, tongue cancer, multiple myeloma, and leukemia. Indications also include organ fibrosis, such as cystic fibrosis, pulmonary fibrosis, cardiac fibrosis, liver fibrosis, and kidney fibrosis. Indications also include antibacterial, including bacteria (various Gram-negative and Gram-positive bacteria) and fungi.

[0053] As used herein, the term "preventing" includes reducing the likelihood of a disease or condition occurring or developing in a patient.

[0054] As used herein, the term "treatment" and other similar synonyms include the following meanings:

[0055] (i) preventing a disease or condition from occurring in a mammal, particularly where such mammal is susceptible to the disease or condition but has not yet been diagnosed as having the disease or condition;

[0056] (ii) inhibiting a disease or condition, i.e. arresting its development;

[0057] (iii) alleviate the disease or condition, that is, cause regression of the disease or condition; or

[0058] (iv) alleviating the symptoms caused by the disease or condition.

[0059] As used herein, the term "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" refers to an amount of at least one agent or compound sufficient to relieve to some extent one or more symptoms of the disease or condition being treated after administration. The result can be a reduction and / or alleviation of signs, symptoms or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein required to provide a clinically significant symptom alleviation effect. Techniques such as dose escalation trials can be used to determine the effective amount appropriate for any individual case.

[0060] As used herein, the terms "administering", "administering", "dosing", etc. refer to methods that can deliver a compound or composition to the desired site for biological action. These methods include, but are not limited to, oral routes, transduodenal routes, parenteral injections (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical administration, and rectal administration. Those skilled in the art are familiar with the administration techniques that can be used for the compounds and methods described herein. In a preferred embodiment, the compounds and compositions of the present invention are administered orally, and formulations of the compounds disclosed herein suitable for oral administration can be presented in discrete units, such as tablets, capsules, or cachets, each containing a predetermined amount of active ingredients. In other preferred embodiments, the compounds and compositions of the present invention are injections and powders.

[0061] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0062] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0063] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures.

[0064] In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0065] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0066] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0067] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0068] Dosage forms for topical administration of a compound of this invention include ointments, powders, patches, sprays and inhalants.

[0069] The active ingredients are mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required if necessary. The terms "pharmaceutical combination", "pharmaceutical combination", "combination therapy", "administration of other treatments", "administration of other therapeutic agents" and the like as used herein refer to a pharmaceutical treatment obtained by mixing or combining more than one active ingredient, including fixed and non-fixed combinations of the active ingredients. The term "fixed combination" refers to the simultaneous administration of at least one compound described herein and at least one synergistic agent to a patient in the form of a single entity or a single dosage form. The term "non-fixed combination" refers to the simultaneous administration, co-administration or sequential administration of at least one compound described herein and at least one synergistic agent to a patient in the form of separate entities at variable intervals.

[0070] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 25-500 mg, preferably 100-250 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0071] The present invention also provides a method for preparing a pharmaceutical composition, comprising the steps of: mixing a pharmaceutical carrier with the compound of general formula (I) of the present invention or its crystal form, pharmaceutical salt, hydrate or solvate, thereby forming a pharmaceutical composition.

[0072] The present invention also provides a treatment method, which comprises the steps of administering the compound of the present invention, or a pharmaceutically acceptable salt thereof, or administering the pharmaceutical composition of the present invention to a subject in need of treatment, for inhibiting HSP90.

[0073] The representative compound number, name, structure and high-resolution mass spectrometry results are shown in Table 1 below:

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101] Preparation method of compound of general formula (I)

[0102] It should be noted that the various materials and reagents used in the following examples are commonly used materials and reagents in the art and can be obtained commercially; the amounts mentioned in the following examples are molar amounts.

[0103] Synthesis route 1:

[0104] Preparation Example 1 Preparation Method of Intermediate 1 (Product 6 in this Preparation Example)

[0105]

[0106] Methods and reagents: A: BF3OEt2, AcOH, 90℃, 16h; B: dry N,N-dimethylformamide (dry DMF), BnBr, K2CO3, Ar, 140℃, 24h; C: NaH, dry tetrahydrofuran (dry THF), diethyl oxalate, Ar, 0℃, 0.5h, 90℃, 3h, reflux; D: NH2OH.HCl, dry ethanol (dry CH3CH2OH), 90℃, 4h, reflux; E: i): tetrahydrofuran (THF), H2O, LiOH, RT, 24h; ii) EtNH2.HCl, HOBT, EDCI, ET3N, dry dichloromethane (dryDCM), 0℃, 0.5h, RT 24h; F: CAN, NIS, dry acetonitrile (dry CH3CN), RT, 24h.

[0107] The specific steps include:

[0108] The raw material compound (1 times amount) was dissolved in etherified boron trifluoride (15 times amount), and acetic acid (5 times amount) was added to a round-bottom flask, and the gas was received by a balloon. The reaction was stirred at 90°C for 16 hours. After cooling to room temperature, an appropriate amount of 10% NaOAC solution was added, and the mixture was stirred at room temperature for half an hour. The mixture was extracted with ethyl acetate, washed three times with a saturated sodium chloride aqueous solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. After column chromatography purification (petroleum ether: ethyl acetate = 30:1), the pure product 1 was obtained as a brown solid.

[0109] Compound 1 (1 times amount) was dissolved in anhydrous dichloromethane, and anhydrous potassium carbonate (2.2 times amount) and benzyl bromide (2.2 times amount) were added to a round-bottom flask at room temperature, argon was replaced, and heated and stirred at 140°C overnight. After cooling to room temperature, water was added to the reaction solution, and solids precipitated. The filter cake was washed with water three times and petroleum ether three times in sequence, and purified by column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain pure khaki compound 2.

[0110] Compound 2 (1 times amount) was dissolved in anhydrous N,N-dimethylformamide, and the mixture was slowly added dropwise to a two-necked bottle containing sodium hydride (4.0 times amount) after argon was replaced, and the mixture was stirred at room temperature for half an hour, and diethyl oxalate (1.5 times amount) was slowly added, and the mixture was stirred and refluxed at 90°C for 3 hours. The mixture was cooled to room temperature, and the pH was adjusted to weak acid (PH=5) with acetic acid, and water was added to the mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed three times with a saturated sodium chloride solution, and the ethyl acetate layer was dried with anhydrous sodium sulfate for half an hour, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1) to obtain the pure product 3 as a bright yellow solid.

[0111] Compound 3 (1 times amount) was dissolved in anhydrous ethanol, and hydroxylamine hydrochloride (1.5 times amount) was added to the reaction solution at room temperature, and refluxed at 90°C for 4h. The reaction solution was cooled to room temperature, concentrated, and water was added. The organic layer was extracted with ethyl acetate, and the organic layer was washed with saturated sodium chloride. The ethyl acetate layer was dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain the pure product 4 as a yellow compound.

[0112] Compound 4 (1 times the amount) was added to a round-bottom flask, and a mixture of tetrahydrofuran, 1M lithium hydroxide and water (v / v / v=2:1:1) was added. Stir at room temperature overnight. The reaction solution was concentrated, water was added to the concentrate, the pH was adjusted to acidic with 1mol / L HCl, extracted with ethyl acetate, washed with a saturated sodium chloride aqueous solution, and dried over anhydrous sodium sulfate. The dried organic liquid was filtered, concentrated, dried in a drying oven, and placed in a round-bottom flask, dissolved in anhydrous dichloromethane, and ethylamine hydrochloride (1.5 times the amount), HOBT (1.5 times the amount), EDCI (2 times the amount), and triethylamine (3.5 times the amount) were added to the reaction solution in turn under an ice bath, reacted at 0°C for half an hour, the ice bath was removed, stirred at room temperature overnight, the reaction solution was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain product 5 as a yellow compound.

[0113] NIS (2.0 times amount) and CAN (0.05 times amount) were added to anhydrous acetonitrile solution of compound 5 (1 times amount), stirred at room temperature overnight, and protected from light. Acetonitrile was evaporated, extracted with ethyl acetate, and the organic layer was washed three times with saturated sodium chloride solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) gave pure product 6 as a white compound.

[0114] Preparation Example 2 Preparation Method of Intermediate 2 (Final Product in This Preparation Example)

[0115]

[0116] Methods and reagents: A: NaHCO3, Pd(PPh3)2Cl2, N,N-dimethylformamide (DMF), H2O, Ar, 80℃, 24h; B: BCl3, anhydrous dichloromethane (dry DCM), -78℃, 1h, RT, 2h; C: NHOK, RT, 24h.

[0117] The specific steps include:

[0118] Compound 6 (1 times the amount), compound 7 (1.5 times the amount), sodium bicarbonate (3 times the amount) and bistriphenylphosphine palladium dichloride (0.01 times the amount) were placed in a two-necked bottle. The air in the bottle was replaced with argon. A mixture of N,N-dimethylformamide and water (v / v=5:1) that had been evacuated with argon was added. The mixture was reacted at 90°C overnight. After cooling to room temperature, the reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate until no fluorescence was washed out. The collected filtrate was washed 3 times with a saturated sodium chloride aqueous solution, the ethyl acetate layer was dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether: ethyl acetate=15:1) to obtain the pure product 8 as a yellow solid.

[0119] Compound 8 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) gave pure product 9 as a light yellow compound.

[0120] The final compound 9 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 65%: 35%) to obtain a pure white solid final product.

[0121] Preparation Example 3 A method for preparing a compound of the present invention, wherein R2 of the compound is para-O-CH2-Ph-CONHOH, L is free of any substituent, and X is O:

[0122]

[0123] Methods and reagents: A: K2CO3, CH3CN, RT, 24h; B: BCl3; dry dichloromethane (dry DCM), -78℃, 1h, RT, 2h; C: NaCO3, Pd(PPh3)2Cl2, dioxane, H2O, Ar, 80℃, 24h; D: NHOK, RT, 24h.

[0124] The specific steps include:

[0125] A: Compound 10 (1 times amount), compound 11 (1.2 times amount), and anhydrous potassium carbonate (3 times amount) were placed in a round-bottom flask, and an appropriate amount of acetonitrile was added to dissolve them, and stirred at room temperature overnight. The acetonitrile was evaporated, and double distilled water was added, and ethyl acetate was extracted. The mixture was washed three times with saturated sodium chloride solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. After column chromatography purification (petroleum ether: ethyl acetate = 20:1), the pure product 12 was obtained as a white solid.

[0126] Compound 6 (1 times the amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times the amount) was slowly added dropwise at -78°C, stirred for 1 hour, and then stirred for 2 hours at room temperature. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. The residue was dissolved in dichloromethane and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain the pure product 13 as a light yellow oily compound.

[0127] Compound 13 (1 times the amount), compound 12 (2 times the amount), anhydrous sodium carbonate (3 times the amount) and bistriphenylphosphine palladium dichloride (0.02 times the amount) were placed in a two-necked bottle. The air in the bottle was replaced with argon. A mixture of dioxane and water (v / v = 4:1) that had been evacuated with argon was added. The mixture was reacted at 80°C overnight. After cooling to room temperature, the reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate until no fluorescence was washed out. The filtrate was collected and washed 3 times with a saturated sodium chloride aqueous solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. The pure product 14 was obtained as a white solid by high performance liquid chromatography (methanol: water = 70%: 30%).

[0128] The final compound 14 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 65%: 35%) to obtain a pure white solid final product.

[0129] The preparation method of the product in which R2 is meta-O-CH2-Ph-CONHOH is the same as the preparation method of para-O-CH2-Ph-CONHOH, except that the hydroxyl group of compound 10 in the raw material is meta-position.

[0130] Preparation Example 4 A method for preparing a compound of the present invention, wherein R2 of the compound is para-CH=CH-CONHOH, L is free of any substituent, and X is O:

[0131]

[0132] Methods and reagents: A: Na2CO3, Pd(PPh3)2Cl2, DMF, H2O, Ar, 80℃, 5h; B: BCl3, dry DCM, -78℃, 1h, RT, 2h; C: NHOK, RT, 24h.

[0133] The specific steps include:

[0134] Compound 6 (1 times the amount), compound 15 (1.5 times the amount), sodium bicarbonate (3 times the amount) and bistriphenylphosphine palladium dichloride (0.02 times the amount) were placed in a two-necked bottle. The air in the bottle was replaced with argon. A mixture of N,N-dimethylformamide (DMF) and water (v / v=5:1) that had been evacuated with argon was added. The mixture was reacted at 80°C for 5h. After cooling to room temperature, the reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate until no fluorescence was washed out. The collected filtrate was washed 3 times with a saturated sodium chloride aqueous solution, the ethyl acetate layer was dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether: ethyl acetate=10:1) to obtain the pure product 16 as a yellow oily compound.

[0135] Compound 16 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) gave pure product 17 as a light yellow solid compound.

[0136] The final compound 17 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 65%: 35%) to obtain a pure product as an orange solid.

[0137] The preparation method of the product in which R2 is meta-CH=CH-CONHOH is the same as that of para-CH=CH-CONHOH, except that the -CH=CH-COOCH3 of compound 15 in the raw material is meta-position.

[0138] Preparation Example 5 A method for preparing a compound of the present invention, wherein R2 of the compound is para-CONHOH, L is an alkyl group, and X is O:

[0139]

[0140] Methods and reagents: A: sodium carbonate, tetrakistriphenylphosphine palladium, dioxane, water, argon, 90℃, 24h; B: boron trichloride, anhydrous dichloromethane, -78℃, 1h, room temperature, 2h; C: potassium hydroxylamine, anhydrous methanol, room temperature, overnight.

[0141] The specific steps include:

[0142] Compound 6 (1 times the amount), compound 18 (1.5 times the amount), anhydrous sodium carbonate (3 times the amount) and tetrakistriphenylphosphine palladium (0.02 times the amount) are placed in a double-necked bottle. The air in the bottle is replaced with argon. A mixture of dioxane and water (v / v=5:1) that has been evacuated with argon is injected. The mixture is reacted at 90°C overnight. After cooling to room temperature, the reaction solution is filtered through diatomaceous earth, and the filter cake is rinsed with ethyl acetate until no fluorescence is washed out. The collected filtrate is washed 3 times with a saturated sodium chloride aqueous solution, the ethyl acetate layer is dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether: ethyl acetate=10:1) to obtain pure product 19 as a white solid compound.

[0143] Compound 19 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and then continued to react for 2 hours after being transferred to room temperature. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) gave the pure product 20 as a light yellow solid compound.

[0144] The final compound 20 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 65%: 35%) to obtain a pure product as a light yellow solid final product.

[0145] The preparation method of the final product in which R2 is meta-CONHOH is the same as that of para-CONHOH, except that the -COOCH3 of compound 18 in the raw material is meta-position.

[0146] Preparation Example 6 A method for preparing a compound of the present invention, wherein R2 of the compound is para-O-Ph-CONHOH, L is free of any substituent, and X is O:

[0147]

[0148] Methods and reagents: A: NaHCO3, Pd(PPh3)2Cl2, DMF, H2O, Ar, 80℃, 24h; B: Cu(OAc)2, pyridine, anhydrous dichloromethane, Molecular sieves, oxygen, RT, 24h; C: BCl3, dry dichloromethane (dry DCM), -78℃, 1h, RT, 2h; D: NHOK, anhydrous methanol, RT, 24h.

[0149] The specific steps include:

[0150] (i) Compound 6 (1 times the amount), compound 10 (2 times the amount), sodium bicarbonate (3 times the amount) and bistriphenylphosphine palladium dichloride (0.02 times the amount) are placed in a double-necked bottle. The air in the bottle is replaced with argon. A mixture of N,N-dimethylformamide and water (v / v=5:1) that has been evacuated with argon is added. The mixture is reacted at 80°C overnight. After cooling to room temperature, the reaction solution is filtered through diatomaceous earth, and the filter cake is rinsed with ethyl acetate until no fluorescence is washed out. The collected filtrate is washed three times with a saturated sodium chloride aqueous solution, the ethyl acetate layer is dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether: ethyl acetate=6:1) to obtain the pure product 21 as a yellow solid compound.

[0151] Compound 21 (1 times the amount), compound 7 (1.1 times the amount), copper acetate (1.1 times the amount) and a certain amount The molecular sieve was placed in a double-necked bottle. The air in the bottle was replaced with oxygen. Pyridine (5 times the amount) and anhydrous dichloromethane were added through a syringe. The mixture was stirred and reacted overnight at room temperature. The reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate until no fluorescence was washed out. The collected filtrate was washed 3 times with a saturated sodium chloride aqueous solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. After column chromatography purification (petroleum ether: ethyl acetate = 8:1), the pure product 22 was obtained as a yellow solid compound.

[0152] Compound 22 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) gave pure product 23 as a light yellow solid compound.

[0153] The final compound 23 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 65%: 35%) to obtain a pure white solid final product.

[0154] The preparation method of the product in which R2 is meta-O-Ph-CONHOH is the same as the preparation method of the product in which R2 is para-O-Ph-CONHOH, except that the hydroxyl group in compound 10 is meta-position.

[0155] Compound 21 (1 times the amount), compound 24 (1.1 times the amount), copper acetate (1.1 times the amount) and a certain amount The molecular sieve was placed in a double-necked bottle. The air in the bottle was replaced with oxygen. Pyridine (5 times the amount) and anhydrous dichloromethane were added by syringe. The mixture was stirred and reacted overnight at room temperature. The reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate until no fluorescence was washed out. The filtrate was collected and washed with saturated sodium chloride aqueous solution 3 times, and the ethyl acetate layer was dried over anhydrous sodium sulfate. After column chromatography purification (petroleum ether: ethyl acetate = 8:1), the pure product 25 was obtained as a yellow solid compound.

[0156] Compound 25 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) gave pure product 26 as a light yellow solid compound.

[0157] The final compound 26 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 65%: 35%) to obtain a pure white solid final product.

[0158] The preparation method of the product in which R2 is meta-O-Ph-CH=CH-CONHOH is the same as the preparation method of the product in which R2 is para-O-Ph-CH=CH-CONHOH, except that the hydroxyl group in compound 21 is meta-position.

[0159] Preparation Example 7 A method for preparing a compound of the present invention, wherein R2 of the compound is para-CONHOH, L is an alkyl, alkenyl or alkynyl group, and X is O:

[0160]

[0161] Methods and reagents: A: CuI, Pd(PPh3)2Cl2, Et3N, MeCN, Ar, 70℃, 12h; B: BCl3, dry DCM, -78℃, 1h, RT, 2h; C: NHOK, RT, 24h; D: Pd(OAc)2, PPh3, K2CO3, DMF, H2O; E: H2, 10% Pd / C, CH3OH, RT, 1h.

[0162] The specific steps include:

[0163] Compound 6 (1 times the amount), compound 27 (1.2 times the amount), cuprous iodide (0.05 times the amount), and dichlorobistriphenylphosphine palladium (0.05 times the amount) were placed in a double-necked bottle. The air in the bottle was replaced with argon. Anhydrous triethylamine (3 times the amount) and acetonitrile bubbled with argon were added through a syringe. The mixture was stirred and reacted at 70°C for 12 hours. The reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate until no fluorescence was washed out. The collected filtrate was washed three times with a saturated sodium chloride aqueous solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. After column chromatography purification (petroleum ether: ethyl acetate = 10:1), the pure product 28 was obtained as a light yellow solid.

[0164] Compound 28 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) gave pure product 29 as a light yellow solid.

[0165] The final compound 29 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 60%: 40%) to obtain a pure product as a light yellow solid.

[0166] Compound 6 (1 times the amount), compound 30 (1.2 times the amount), palladium acetate (0.05 times the amount) and triphenylphosphine (0.1 times the amount) were placed in a two-necked bottle. The air in the bottle was replaced with argon. A mixture of N,N-dimethylformamide and water (v / v=5:1) that had been evacuated with argon was added. The mixture was stirred and reacted overnight at 100°C. The reaction solution was filtered through diatomaceous earth, and the filter cake was rinsed with ethyl acetate until no fluorescence was washed out. The collected filtrate was washed three times with a saturated sodium chloride aqueous solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. After column chromatography purification (petroleum ether: ethyl acetate=10:1), the pure product 31 was obtained as a light yellow solid.

[0167] Compound 31 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (dichloromethane: methanol = 80: 1) gave pure product 32 as a light yellow solid compound.

[0168] The final compound 32 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain the final crude product, which was purified by high performance liquid chromatography (methanol: water = 65%: 35%) to obtain a pure off-white solid final product.

[0169] Compound 31 (1 times amount) was dissolved in methanol solution, 10% palladium carbon (0.1 times amount) was added, the air in the bottle was replaced with hydrogen, the reaction was stirred at room temperature for 1 hour, the reaction solution was filtered through diatomaceous earth, and the filter cake was washed with methanol until no fluorescence was washed out. Purification by silica gel column chromatography (dichloromethane: methanol = 40: 1) gave pure product 33 as a light yellow solid compound.

[0170] The final compound 33 was dissolved in anhydrous methanol solution of potassium hydroxylamine and stirred at room temperature overnight. After removing methanol under reduced pressure, double distilled water was added to dissolve the residue, and the pH value was adjusted to 6-7 with 1M HCl solution. The generated solid was filtered and washed with double distilled water 4 times to obtain a crude final product, which was purified by high performance liquid chromatography (methanol: water = 70%: 30%) to obtain a pure off-white solid final product.

[0171] The preparation method of R2 as a meta-substituent is consistent with the preparation method of R2 as a para-substituent, as long as the -COOCH3 in compounds 27 and 30 is in the meta-position.

[0172] Preparation Example 8 A method for preparing a compound of the present invention, wherein R2 of the compound is para-SO2CH3, para-SOCH3, para-PO(CH3)2 or para-NO2, L is free of any substituent, and X is O:

[0173]

[0174] Methods and reagents: A: NaHCO3, Pd(PPh3)2Cl2, DMF, H2O, Ar, 90℃, 24h; B: BCl3, dry DCM, -78℃, 1h, RT, 2h.

[0175] The specific steps include:

[0176] Compound 6 (1 times the amount), compound 34 or 37 or 40 or 43 (2 times the amount), sodium bicarbonate (3 times the amount) and bistriphenylphosphine palladium dichloride (0.01 times the amount) are placed in a double-necked bottle. The air in the bottle is replaced with argon. A mixture of N,N-dimethylformamide and water (v / v=5:1) that has been evacuated with argon is added. The mixture is reacted at 80°C overnight. After cooling to room temperature, the reaction solution is filtered through diatomaceous earth, and the filter cake is rinsed with ethyl acetate until no fluorescence is washed out. The collected filtrate is washed 3 times with a saturated sodium chloride aqueous solution, the ethyl acetate layer is dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether: ethyl acetate=8:1) to obtain pure product 35 or 38 or 41 or 44 as a yellow solid compound.

[0177] Compound 35 or 38 or 41 or 44 (1 times the amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times the amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) gave pure product 36 or 39 or 42 or 45 as a white solid.

[0178] Preparation Example 9 A method for preparing a compound of the present invention, wherein R2 of the compound is meta-SO2CH3, meta-SOCH3, meta-PO(CH3)2 or meta-NO2, L is free of any substituent, and X is O:

[0179]

[0180] Methods and reagents: A: NaHCO3, Pd(PPh3)2Cl2, DMF, H2O, Ar, 90℃, 24h; B: BCl3, anhydrous dichloromethane, -78℃, 1h, RT, 2h.

[0181] The specific steps include:

[0182] Compound 6 (1 times the amount), compound 46 or 49 or 52 or 55 (2 times the amount), sodium bicarbonate (3 times the amount) and bistriphenylphosphine palladium dichloride (0.1 times the amount) are placed in a double-necked bottle. The bottle is replaced with argon. A mixture of N,N-dimethylformamide and water (v / v=5:1) that has been evacuated with argon is added. The mixture is reacted at 80°C overnight. After cooling to room temperature, the reaction solution is filtered through diatomaceous earth, and the filter cake is rinsed with ethyl acetate until no fluorescence is washed out. The collected filtrate is washed 3 times with a saturated sodium chloride aqueous solution, the ethyl acetate layer is dried over anhydrous sodium sulfate, and purified by column chromatography (petroleum ether: ethyl acetate=8:1) to obtain pure product 47 or 50 or 53 or 56 as a yellow solid compound.

[0183] Compound 47 or 50 or 53 or 56 (1 times amount) was dissolved in anhydrous dichloromethane, and then boron trichloride (4 times amount) was slowly added dropwise at -78°C, stirred for 1 hour, and reacted at room temperature for 2 hours. The reaction solution was concentrated, then dissolved in methanol, and concentrated again, and repeated three times. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) gave pure product 48 or 51 or 54 or 57 as a yellow solid.

[0184] Synthesis route 2:

[0185] Preparation Example 10 Preparation method of intermediate 3

[0186]

[0187] Methods and reagents: A: BF3OEt2, AcOH, 90℃, 16h; B: anhydrous N,N-dimethylformamide, BnBr, K2CO3, Ar, 140℃, 24h; C: NaH, anhydrous tetrahydrofuran, diethyl oxalate, argon, 0℃, 0.5h, 90℃, 3h, reflux; D: hydrazine hydrate, ethanol, 90℃, 4h, reflux; E: i): THF, H2O, LiOH, RT, 24h; ii) ethylamine hydrochloride, HOBt, EDCI, triethylamine, anhydrous dichloromethane, 0℃, 0.5h, room temperature, 24h; F: CAN, NIS, anhydrous acetonitrile, room temperature, 24h;

[0188] The specific steps include:

[0189] The raw material compound 6-position R-substituted resorcinol (1 times amount) in the preparation example of synthetic route 1 was dissolved in etherified boron trifluoride (15 times amount), and acetic acid (5 times amount) was added to a round-bottom flask, and the gas was received by a balloon. The reaction was stirred at 90°C for 16 hours. After cooling to room temperature, an appropriate amount of 10% NaOAC solution was added, and the mixture was stirred at room temperature for half an hour. The mixture was extracted with ethyl acetate, washed three times with a saturated sodium chloride aqueous solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. After column chromatography purification (petroleum ether: ethyl acetate = 30:1), the pure product 1 was obtained as a brown solid.

[0190] Compound 1 (1 times amount) was dissolved in anhydrous dichloromethane, and anhydrous potassium carbonate (2.2 times amount) and benzyl bromide (2.2 times amount) were added to a round-bottom flask at room temperature, argon was replaced, and heated and stirred at 140°C overnight. After cooling to room temperature, water was added to the reaction solution, and solids precipitated. The filter cake was washed with water three times and petroleum ether three times in sequence, and purified by column chromatography (petroleum ether: ethyl acetate = 20:1) to obtain pure khaki compound 2.

[0191] Compound 2 (1 times amount) was dissolved in anhydrous N,N-dimethylformamide, and the mixture was slowly added dropwise to a two-necked bottle containing sodium hydride (4.0 times amount) after argon was replaced, and the mixture was stirred at room temperature for half an hour, and diethyl oxalate (1.5 times amount) was slowly added, and the mixture was stirred and refluxed at 90°C for 3 hours. The mixture was cooled to room temperature, and the pH was adjusted to weak acid (PH=5) with acetic acid, and water was added to the mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed three times with a saturated sodium chloride solution, and the ethyl acetate layer was dried with anhydrous sodium sulfate for half an hour, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1) to obtain the pure product 3 as a bright yellow solid.

[0192] Compound 3 (1 times amount) was dissolved in anhydrous ethanol, and hydrazine hydrate (1.5 times amount) was added to the reaction solution at room temperature, and refluxed at 90°C for 4 hours. The reaction solution was cooled to room temperature, concentrated, and water was added. The organic layer was extracted with ethyl acetate, and the organic layer was washed with saturated sodium chloride. The ethyl acetate layer was dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain the pure product 58 as a yellow compound.

[0193] Compound 58 (1 times amount) was added to a round-bottom flask, and a mixture of tetrahydrofuran, lithium hydroxide and water (v / v / v = 2:1:1) was added. Stir at room temperature overnight. The reaction solution was concentrated, water was added to the concentrate, the pH was adjusted to acidic with 1 mol / L HCl, extracted with ethyl acetate, washed with a saturated sodium chloride aqueous solution, and dried over anhydrous sodium sulfate. The dried organic solution was filtered, concentrated, dried in a drying oven, and placed in a round-bottom flask, dissolved in anhydrous dichloromethane, and ethylamine hydrochloride (1.5 times amount), HOBt (1.5 times amount), EDCI (2 times amount), and triethylamine (3.5 times amount) were added to the reaction solution in turn under an ice bath, reacted at 0°C for half an hour, the ice bath was removed, stirred at room temperature overnight, the reaction solution was concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain product 59 as a yellow compound.

[0194] NIS (2.0 times amount) and CAN (0.05 times amount) were added to anhydrous acetonitrile solution of compound 5 (1 times amount), stirred at room temperature overnight, and protected from light. Acetonitrile was evaporated, extracted with ethyl acetate, and the organic layer was washed three times with saturated sodium chloride solution, and the ethyl acetate layer was dried over anhydrous sodium sulfate. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) gave pure product 60 as a brown compound.

[0195] The synthesis methods of all the remaining target compounds of the pyrazole series are completely consistent with the synthesis methods of the various end products of the first oxazole series mentioned above, and will not be repeated here.

[0196] Activity test results of representative compounds:

[0197] (I) Activity inhibition experiment

[0198] First, the single concentration inhibition rate of 112 representative compounds on HSP90 was tested, and the administration concentration was 200nM. The specific steps included: 100mM TrisCl pH7.4, 20mM KCl, 6mM MgCl2, 5ug / ml BSA, 25nM full-length Hsp90, 10nM VER-51001 (fluorescent probe) were mixed to a total volume of 50μL; the mixture was equilibrated at room temperature for 30 minutes in the dark using a 384-well plate; then 200nM of the compound was added and equilibrated for another 30 minutes in the dark at room temperature (the final concentration of DMSO was 2%); fluorescence polarization was measured using a Fusion plate reader (Perkin Elmer; absorption wavelength 485nm, emission wavelength 535nm), and the inhibition rate was calculated. The results are shown in Figures 1(a) and 1(b). It can be seen from the results that the inhibition rate of all compounds on HSP90 at a concentration of 200nM was higher than 85%. It was demonstrated that 112 representative compounds had extremely high HSP90 inhibitory activity.

[0199] Then, the proliferation inhibitory activity of 112 compounds on A549 lung cancer cells and lung fibroblasts HLF-1 was tested. The specific steps included: using culture medium containing 10% fetal calf serum to prepare a single cell suspension, inoculating 10,000 cells per well into a 96-well plate with a volume of 200 microliters per well. After adding compounds of different concentrations, incubate for 72 hours. Add 20 microliters of MTT solution (5 mg / ml prepared with PBS) to each well. Continue incubation for 4 hours, terminate the culture, and carefully discard the culture supernatant in the well. For suspended cells, centrifuge and then discard the culture supernatant in the well. Add 150 microliters of DMSO to each well and oscillate for 10 minutes to fully dissolve the crystals. Select a wavelength of 490nm, measure the light absorbance of each well on an enzyme-linked immunosorbent assay, record the results, draw a cell growth curve with time as the horizontal axis and absorbance as the vertical axis, and calculate the IC 50 The results are shown in Figure 2(a) and Figure 2(b).

[0200] From the results, it can be seen that 112 representative compounds showed strong proliferation inhibition activity on A549 cells and lung fibroblast HLF-1 cells. 50 The values ​​are all at the nM level, demonstrating that this class of compounds has the potential to be used as anticancer and pulmonary fibrosis drugs.

[0201] (II) A representative compound HH-69 was selected to test its inhibitory activity against the A549 cell mouse tumor model. The specific steps included: BALB / c mice were randomly divided into three groups (n=10 mice in each group): Group 1, saline and dimethyl sulfoxide (5%, v / v); Group 2 was given 10 mg / kg of HH-69 and saline and dimethyl sulfoxide (5%, v / v); Group 3 was the drug administration group, 20 mg / kg of HH-69 and saline and dimethyl sulfoxide (5%, v / v), once every three days, intraperitoneally for 18 days; after 18 days, the mice were killed, the tumors were removed, and the tumor volume (length × width) was calculated. 2 ÷2), the statistical results are as follows Figure 3 shown.

[0202] It can be seen from the results that compound HH-69 can significantly inhibit tumor growth at both low and high doses by intraperitoneal injection for 18 days.

[0203] (III) In addition, the compound HH-69 was tested for reversing bleomycin-induced pulmonary fibrosis in mice. The specific steps included: CBA / J mice were randomly divided into three groups, with 6 mice in each group; the first group was a normal mouse group, which was orally administered with normal saline daily; the second group was a bleomycin-induced fibrosis group, in which bleomycin was administered via tracheal injection (2.5 mg / kg), the wound was sutured after the injection, and the mice were allowed to stand upright for 3 minutes to allow bleomycin to be evenly distributed in the lungs. After three days, pulmonary fibrosis lesions were formed; the third group was an HH-69 administration group, and bleomycin was administered 3 days after modeling, with a dose of 10 mg / kg, administered intraperitoneally once a day for 14 days; after 14 days, the mice were dissected, the lung tissues were removed, sliced, and HE-stained, and finally observed under a microscope. The results are as follows: Figure 4 As shown, A is a normal mouse, B is a bleomycin group, and C is a bleomycin + HH-69 group. From the results, it can be seen that compound HH-69 can reverse bleomycin-induced mouse pulmonary fibrosis.

[0204] (IV) Finally, according to literature reports, HSP90 inhibitors have antibacterial activity (J. Med. Chem. 2022, 65, 7, 5539–5564), so we continued to test whether compound HH-69 could inhibit the growth of fungi, using fluconazole as a positive control drug.

[0205] The specific steps include: prepare 10 test tubes. Add 1mL of drug to the first test tube and mix well, pipette 1mL into the second test tube and mix well. Pipet 1mL into the second test tube and add it to the third test tube, and so on, when it comes to the tenth test tube, pipette out 1mL and discard it. Add 1mL of bacterial solution (10000fu / ml) to each of the 10 test tubes and mix well, put them in an incubator and culture for 24 hours, take them out to observe which test tube is still clear, and the drug concentration of that test tube is the MIC. The experimental results are shown in Table 2 below:

[0206]

[0207] From the results, it can be seen that compound HH-69 exhibited strong inhibitory activity against both bacteria, and its activity was significantly better than that of fluconazole.

[0208] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention and do not constitute a limitation on the content of the present invention. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof: in: R1 is selected from the group consisting of alkyl, aryl, methylene aryl and halogen; R2 is selected from the group consisting of para- or meta-O-Ph-CONHOH, para- or meta-O-CH2-Ph-CONHOH, para- or meta-O-Ph-CH=CH-CONHOH, para- or meta-CONHOH, para- or meta-CH=CH-CONHOH, para- or meta-SOCH3, para- or meta-SO2CH3, para- or meta-PO(CH3)2, and para- or meta-NO2; L is absent of any substituent; X is O or NH.

2. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof: in: R1 is selected from the group consisting of alkyl, aryl, methylene aryl and halogen; R2 is selected from the group consisting of para- or meta-O-Ph-CONHOH, para- or meta-O-CH2-Ph-CONHOH, para- or meta-O-Ph-CH=CH-CONHOH, para- or meta-CONHOH, para- or meta-CH=CH-CONHOH, para- or meta-SOCH3, para- or meta-SO2CH3, para- or meta-PO(CH3)2, and para- or meta-NO2; L is selected from the group consisting of alkyl, vinyl, and ethynyl; X is O or NH.

3. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound is selected from 4. A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: The compound is selected from 5. A pharmaceutical composition comprising an effective amount of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable excipient or a pharmaceutically acceptable carrier.

6. Use of the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, and the pharmaceutical composition according to claim 5, in a medicament for preventing and / or treating lymphoma, sarcoma, melanoma, lung cancer, liver cancer, breast cancer, ovarian cancer, prostate cancer, bladder cancer, pancreatic cancer, thyroid cancer, laryngeal cancer, tongue cancer, multiple myeloma, leukemia or cystic fibrosis, pulmonary fibrosis, cardiac fibrosis, liver fibrosis, or kidney fibrosis.

7. Use of the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, or the pharmaceutical composition according to claim 5, in an antibacterial drug.

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