Hydroximic acid compounds containing tetrahydrobenzo[4,5]furano[2,3-c]pyridine, preparation method therefor, and uses thereof

Abstract

Provided are compounds of formula (1) and / or formula (2), and / or pharmaceutically acceptable salts thereof, and / or racemic mixtures, hydrates, solvates, prodrugs, enantiomers, diastereomers and tautomers thereof, belonging to the technical field of medicines. Also provided are pharmaceutical compositions containing the compounds of formula (1) and / or formula (2), and the uses of the compounds of formula (1) and / or formula (2) and / or the pharmaceutically acceptable salts thereof, and / or the racemic mixtures, hydrates, solvates, prodrugs, enantiomers, diastereomers and tautomers thereof, and / or the pharmaceutical compositions in preparing histone deacetylase inhibitors and drugs used for the treatment and / or prevention of diseases responsive to histone deacetylases.

Description

Hydroxamic acid compound containing tetrahydrobenzo[4,5]furan[2,3-C]pyridine, preparation method and application thereof Technical Field

[0001] The present invention relates to the field of biotechnology, and more specifically, to a 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine-containing hydroxamic acid compound and a preparation method thereof, as well as use of the compound as a histone deacetylase inhibitor in treating diseases associated with histone deacetylase. Background Art

[0002] Due to its high incidence and mortality rates, cancer has become the leading cause of death worldwide. Cancer research has found that epigenetics plays an important regulatory role in the occurrence, development, invasion and metastasis of tumors. Post-synthesis processing of proteins is an important process of physiological regulation, including methylation, acetylation, phosphorylation, etc., which can enable individual proteins to exist in multiple functional states and reversibly transfer from one state to another. Protein acetylation occurs on lysine residues, and the acetylation modification of histones can regulate gene transcription. Studies have found that the expression levels of histone deacetylases are abnormally increased in various cancers. In addition, the levels of protein acetylation and deacetylation are closely related to inflammation. Because histone deacetylases are key enzymes that regulate the level of protein acetylation in the body, histone deacetylases are an important target in the treatment of cancer and inflammation research.

[0003] Histone deacetylase catalyzes the removal of acetyl groups from histone lysine residues, imparting a positive charge to the lysine residues. This allows the lysine residues to tightly wrap around DNA, resulting in chromosome contraction and the inhibition of gene expression. Numerous studies have shown that histone deacetylase levels are elevated in various cancers and are often associated with poor prognosis. Studies have found that histone deacetylase inhibitors can inhibit angiogenesis in the tumor microenvironment and induce programmed cell death in tumor cells, while having no significant inhibitory effect on normal cells. Therefore, HDAC inhibitors are a novel anti-tumor drug.

[0004] Summary of the Invention

[0005] The present invention aims to provide a 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine-containing hydroxamic acid compound, a preparation method and application thereof, and the application of the compound as a histone deacetylase inhibitor in the prevention and / or treatment of tumors.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] The present invention provides a compound represented by formula (I) or formula (II) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs;

[0008] Among them, R 1 、R 2 、R 3 、R 4 are the same or different and are each independently selected from one or more of hydrogen, halogen, nitro, cyano, amino, hydroxyl, hydroxymethyl, hydroxyethyl, thiol, carboxyl, ester, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, aromatic heteroyl, alkenyl, alkynyl, monosubstituted alkylamino, disubstituted alkylamino, alkoxy, alkylcarbonyloxy, alkylcarbonylamino, cycloalkylcarbonylamino, heteroarylcarbonylamino, aminocarbonyl, alkoxyformamido, alkylthio, hydroxyalkoxy, sugar residue, sulfonic acid, phosphoric acid, polyhydroxyalkoxycarbonyl, carboxyalkoxy, carboxyalkylformyloxy, or R 2 R 3 forms a ring structure of a substituted or unsubstituted aryl group, or forms a ring structure of a substituted or unsubstituted heteroaryl group, or forms a ring structure of a substituted or unsubstituted alkyl group; R 5 For H, Where n is a natural number between 0 and 2. represents the junction site;

[0009] L in formula (1) and / or formula (2) is selected from the following:

[0010] Where n is a natural number from 0 to 8. Represents the attachment site of L.

[0011] As a preference, R 1 、R 2 、R 3 、R 4 are the same or different, and are independently selected from hydrogen, C1-C8 alkyl, C1-C8 alkoxy, halogen, hydroxyl, or R 2 、R 3 Connect with R 2 and R 3 The two C's together form a heterocycloalkyl group, wherein the heterocycloalkyl group is a 3-8 membered ring having 1 or 2 selected from N, O, and S;

[0012] R 5 For H, Where n is a natural number between 0 and 2. represents the junction site;

[0013] L in formula (1) and / or formula (2) is selected from the following:

[0014] Where n is a natural number from 0 to 8. Represents the attachment site of L.

[0015] Specifically, the present invention preferably uses the following compounds:

[0016] Unless otherwise indicated, the term "alkyl" as used herein includes branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, including all isomers; "alkene" refers to branched and straight-chain aliphatic hydrocarbon groups containing a carbon-carbon double bond (olefinic bond); and "alkyne" refers to branched and straight-chain aliphatic hydrocarbon groups containing a carbon-carbon triple bond (alkyne bond). Common abbreviations for alkanes, for example, methyl can be represented by "Me" or CH3, ethyl can be represented by "Et" or CH2CH3, propyl can be represented by "Pr" or CH2CH2CH3, and butyl can be represented by "Bu" or CH2CH2CH2CH3.

[0017] The term "carbonyl" refers to an organic functional group composed of two atoms, carbon and oxygen, connected by a double bond (C=O).

[0018] The terms "cycloalkyl" and "cycloalkenyl" refer to hydrocarbon groups having a monocyclic ring system of carbon atoms which may be saturated or unsaturated.

[0019] The term "aryl" refers to aromatic mono- and polycyclic carbocyclic ring systems, wherein the carbon rings in the polycyclic ring system are fused or connected to each other by single bonds. In the case of polycyclic rings, as long as one carbon ring is aromatic, typical aryl groups include phenyl, naphthyl and biphenylene.

[0020] The term "heterocyclyl" refers to a cyclic structure composed of carbon atoms and non-carbon atoms, such as nitrogen, oxygen, and sulfur. The heterocyclyl may be a monocyclic heterocyclyl having 4-8 ring atoms, or a bicyclic heterocyclyl having 7-11 ring atoms. In the bicyclic heterocyclyl, as long as one ring is a heterocycle, the other ring may be aromatic or non-aromatic, containing or not containing heteroatoms. In addition, the bicyclic heterocyclyl may be a paracyclic structure, a spirocyclic structure, or two heterocycles directly connected. Examples of heterocyclyls include, but are not limited to, azetidinyl, pyrrolidinyl, pyrrolinyl, tetrahydrofuranyl, dihydrofuranyl, piperazinyl, piperidinyl, morpholinyl, thiomorpholinyl, tetrahydrothiophenyl, and the like.

[0021] The term "heteroaryl" refers to an aromatic ring group containing 1-4 heteroatoms as ring members. Heteroatoms refer to nitrogen, oxygen, or sulfur. A heteroaryl group can be a monocyclic heteroaryl group having 5-7 ring atoms, or a bicyclic heteroaryl group having 7-11 ring atoms. In the bicyclic heteroaryl group, as long as one ring is a heteroaromatic ring, the other ring can be aromatic or non-aromatic, containing or not containing heteroatoms. In addition, the bicyclic heteroaryl group can be a paracyclic structure, a spirocyclic structure, or two heterocyclic rings directly connected. Examples of heteroaryl groups include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, pyridyl, pyrimidinyl, furanyl, thienyl, indolyl, and the like.

[0022] The term "halogen" (or "halo") refers to fluorine, chlorine, bromine, and iodine (alternatively referred to as fluoro (F), chloro (Cl), bromo (Br), and iodo (I)).

[0023] In the present invention, "C1-C8 alkyl" refers to a straight chain or branched chain alkyl group with a specific number of carbon atoms (1-8), including all isomers, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, etc. "C2-C8 alkenyl" refers to a straight chain or branched chain alkyl group with a specific number of carbon atoms (2-8), including all isomers, including but not limited to vinyl, allyl, etc. "C3-C8 cycloalkyl or cycloalkenyl" refers to a hydrocarbon group with a saturated or unsaturated ring of 3-8 carbon atoms in a monocyclic system, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopropenyl, cyclohexenyl, etc. Similarly, the term "C1-C8 alkyl" refers to a straight chain or branched chain alkyl group with a specific number of carbon atoms (2-8), including all isomers, including but not limited to vinyl, allyl, etc. 10 Alkyl", "C2-C 10 Alkenyl", "C1-C8 carbonyl", "C3-C8 cycloalkenyl", "C6-C 10 Aryl", "C5-C 10 Heterocyclic group", "C5-C 10 "Heteroaryl" and the like have similar meanings.

[0024] Unless otherwise specified, all ranges listed herein are inclusive. For example, "1-4" includes 1, 2, 3, and 4.

[0025] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound of the present invention is acidic, its corresponding salt can be readily prepared from an inorganic or organic base. Salts derived from such inorganic bases include salts of aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganous), potassium, sodium, zinc, and the like. Preferred are salts of ammonium, calcium, magnesium, potassium, and sodium. Salts prepared from organic bases include primary, secondary, and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable organic non-toxic bases that can form salts include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, dicyclohexylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compound of the present invention is basic, its corresponding salt can be easily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. In the present invention, salts with hydrochloric acid, nitric acid, sulfuric acid, phosphoric acid, hydrobromic acid, maleic acid, fumaric acid, tartaric acid, lactic acid, citric acid, acetic acid, methanesulfonic acid, p-toluenesulfonic acid, adipic acid, palmitic acid, tannic acid, lithium, sodium, potassium, calcium, magnesium, or lysine are preferred.

[0026] The term "solvate" refers to a complex of variable stoichiometry formed by a solute (i.e., a compound of Formula I or Formula II) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is referred to as a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, monosesquihydrates, dihydrates, and trihydrates.

[0027] The term "prodrug" is a functional derivative of the compounds of the invention which is readily convertible in vivo into the required compound.

[0028] The present invention includes a pharmaceutical composition comprising the aforementioned 1,2,3,4-tetrahydrobenzo[4,5]furo[2,3-C]pyridine hydroxamic acid compound as an active ingredient and pharmaceutically acceptable excipients. The pharmaceutically acceptable excipients include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, and the like in the pharmaceutical field.

[0029] The pharmaceutical composition of the present invention can be prepared into several dosage forms, such as injection, tablet, powder, granule, capsule, oral solution, ointment, suppository, etc. The drugs in the above dosage forms can be prepared according to conventional methods in the pharmaceutical field.

[0030] The compounds of the present invention and pharmaceutical compositions comprising the compounds of the present invention can be formulated into a form suitable for intravenous, oral, rectal, parenteral, intranasal or transdermal administration, or a form suitable for administration by inhalation or by suppository administration. Oral and intravenous administration are preferred.

[0031] The 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine hydroxamic acid compounds and pharmaceutical compositions thereof of the present invention are used as histone deacetylase preparations in the prevention and / or treatment of diseases associated with abnormal histone deacetylase expression, such as various cancers, including but not limited to lymphoma, bladder cancer, bone cancer, childhood brainstem glioma, adult brain tumor, cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, craniopharyngioma, ependymoma, breast cancer, bronchiolar tumor, central nervous system lymphoma, cervical cancer, chronic lymphocytic leukemia, colon cancer, colorectal cancer, esophageal cancer, eye cancer, gallbladder cancer, gastric cancer, laryngeal cancer, nasopharyngeal cancer, pancreatic cancer, prostate cancer, intestinal cancer, thyroid cancer, etc. In the present invention, preferred cancers are leukemia, multiple myeloma, lymphoma, liver cancer, stomach cancer, breast cancer, pancreatic cancer, lung cancer, colorectal cancer, cervical cancer, nasopharyngeal cancer, laryngeal cancer, esophageal cancer, melanoma, prostate cancer, osteosarcoma, and glioma.

[0032] The preparation of the 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine hydroxamic acid compound adopts one of the following reaction routes:

[0033] Reaction Scheme 1 includes the following processes:

[0034] Compounds 1, 2, and 3a-3g undergo Mannich reaction to generate compounds 4a-4g, which then react with hydroxylamine to obtain compounds 5a-5g;

[0035] Reaction Scheme 2 includes the following steps:

[0036] Compounds 1a, 2, and 3h underwent a Mannich reaction to generate compound 4h, which was then debenzylated with hydrogen under the catalysis of palladium on carbon to obtain compound 6, which then underwent a condensation reaction with the corresponding carboxylic acids to obtain compounds 7a-7c, and finally reacted with hydroxylamine to obtain compounds 8a-8c.

[0037] Reaction Scheme 3 includes the following processes:

[0038] Compounds 1a, 2, and 3i underwent a Mannich reaction to generate compound 4i, which was then substituted with methyl parahydroxybenzoate to obtain an ester intermediate, which reacted with hydroxylamine to obtain compound 8d.

[0039] Reaction Scheme 4 includes the following processes:

[0040] Compounds 1, 2, and 3 undergo a Mannich reaction to produce compound 4 or compound 9, which is then hydrolyzed to the corresponding carboxylic acid compound 10 under the action of lithium hydroxide, and then condensed with methyl p-aminobenzoate to form compounds 11a-11aa, which are finally reacted with hydroxylamine to produce compounds 12a-12aa;

[0041] Reaction Scheme 5 includes the following process:

[0042] Compound 11ab reacts with trifluoromethanesulfonic anhydride to obtain an intermediate, which is then reacted with formic acid under the catalysis of palladium acetate to produce compound 13, which is then reacted with hydroxylamine to obtain compound 12ab;

[0043] Reaction Scheme 6 includes the following process:

[0044] Compound 10 was condensed with the corresponding amine to give compounds 11ac and 11ad, which were then reacted with hydroxylamine to give compounds 12ac and 12ad.

[0045] Beneficial effects of the present invention

[0046] The 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine hydroxamic acid compounds of the present invention all have good anti-tumor cell proliferation activity, and the preparation method is simple to operate and the conditions are mild. DETAILED DESCRIPTION

[0047] The present invention will be further described in detail below with reference to implementation cases.

[0048] Example 1: Preparation of Compound 5a

[0049] Structural formula of compound 5a

[0050] Step A: Preparation of compound 4a

[0051] In a single-necked flask, 1.62 g of 1a (10 mmol, 1 equiv.), 3.03 g of paraformaldehyde (100 mmol, 10 equiv.), and 2.51 g of glycine methyl ester hydrochloride (20 mmol, 2 equiv.) were weighed and thoroughly dissolved in 20 mL of acetic acid. After stirring, the mixture was heated to 60°C and reacted for 3 hours. After TLC confirmed complete conversion, the mixture was cooled to room temperature and neutralized with saturated sodium bicarbonate solution. Ethyl acetate (3 × 100 mL) was added for extraction. The organic phases were combined, washed with saturated brine (1 × 100 mL), and dried over anhydrous sodium sulfate. Silica gel was added and the sample was mixed. Column chromatography was performed to obtain the desired product 4a as a white solid in a 76% yield.

[0052] Step B: Preparation of compound 5a

[0053] 96 mg of lithium hydroxide (4 mmol, 8 equiv.) and hydroxylamine aqueous solution (5 mmol, 10 equiv.) were weighed into a single-necked flask and dissolved in 2 mL of methanol. At -10°C, 138 mg of compound 4a (0.5 mmol, 1 equiv.) was dissolved in 3 mL of methanol and slowly added dropwise to the reaction solution. After stirring at -10°C for 30 minutes, the mixture was slowly warmed to room temperature and stirred for 8 hours. The reaction solution was then dried, 10 mL of water was added, and the pH was adjusted to 3-4 with 2 M hydrochloric acid. The mixture was filtered, and the residue was washed twice with water and once with ethyl acetate. The filter cake was collected and dried to obtain compound 5a as a white solid with a yield of 72%; mp 140.1-143.2°C; R f =0.4 (DCM:MeOH 20:1). 1 H NMR(600MHz,CD3OD)δ7.30(d,J=9.0Hz,1H),7.01(d,J=2.4Hz,1H),6.83(dd,J1=8.4Hz ,J2=2.4Hz,1H),3.81(s,3H),3.76-3.71(m,2H),2.99-2.87(m,2H),2.78-2.66(m,2H). 13 C NMR(125MHz,CD3OD)δ167.8,157.7,155.7,149.2,121.2,118.3,110.7,110.6,95.8,57.6,54.7,50.1,49.5,19.9.HRMS(ESI)m / z Calcd for C 14 H 17 N2O4(M+H) + :277.1183;Found:277.1196.

[0054] According to the preparation method of Example 1, appropriate raw materials were selected to prepare the compounds of Examples 2 to 7.

[0055] Example 2: Preparation of Compound 5b

[0056] Structural formula of compound 5b

[0057] The preparation method is the same as that of Example 1, white solid, yield 80%; mp 142.1-143.6 ℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR (400MHz, DMSO-d6) δ10.29(s,1H),8.62(s,1H),7.25(d,J=8.4Hz,1H),7.03(d,J=2.4Hz,1H),6.74(dd,J1=8.4Hz,J2=2.4Hz ,1H),3.68(s,3H),3.46(s,2H),2.68-2.59(m,2H),2.54-2.45(m,2H),2.42-2.38(m,3H),1.98-1.86(m,2H),1.70-1.59(m,2H). 13 CNMR(100MHz,DMSO-d6)δ169.6,157.5,155.5,150.8,121.4,119.3,111.4,111.3,96.9,56.6,56.0,50.3,49.9,30.6,23.3,20.9.HRMS(ESI)m / z Calcd for C 16 H 21 N2O4(M+H) + :305.1496;Found:305.1520.

[0058] Example 3: Preparation of Compound 5c

[0059] Structural formula of compound 5c

[0060] The preparation method is the same as that of Example 1, light yellow solid, yield 84%; mp 129.1-130.3℃; R f =0.3 (DCM:MeOH 20:1). 1H NMR (400MHz, DMSO-d6) δ7.36(d,J=8.4Hz,1H),7.14(d,J=2.4Hz,1H),6.85(dd,J1=8.4Hz,J2=2.4Hz,1H),3.79(s, 3H),3.56(s,2H),2.78-2.68(m,2H),2.66-2.57(m,2H),2.56-2.48(m,3H),2.08-1.89(m,2H),1.61-1.43(m,4H). 13 C NMR(100MHz, DMSO-d6)δ169.6,157.4,155.4,150.9,121.4,119.3,111.3,96.9,56.9,56.0,50.4,49.9,32.6,26.7,23.5,20.9.HRMS(ESI)m / z Calcd for C 16 H 23 N2O4(M+H) + :319.1652;Found:319.1658.

[0061] Example 4: Preparation of Compound 5d

[0062] The structural formula of compound 5d

[0063] The preparation method is the same as that of Example 1, light yellow solid, yield 78%; mp 157.5-159.0℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR (400MHz, DMSO-d6) δ10.39(s,1H),8.75(s,1H),7.36(d,J=8.4Hz,1H),7.14(d,J=2.4Hz,1H),6.85(dd,J1=8.4Hz,J2=2.4Hz,1H),3.7 9(s,3H),3.56(s,2H),2.81-2.69(m,2H),2.67-2.55(m,2H),2.56-2.50(m,2H),2.07-1.86(m,2H),1.62-1.45(m,4H),1.40-1.22(m,2H). 13 C NMR(100MHz,DMSO-d6)δ169.6,157.4,155.5,150.9,121.4,119.3,111.3,96.8,57.3,56.0,50.4,49.9,32.7,26.9,25.6,20.9.HRMS(ESI)m / z Calcd for C 18H 25 N2O4(M+H) + :333.1809; Found:333.1826.

[0064] Example 5: Preparation of Compound 5e

[0065] Structural formula of compound 5e

[0066] The preparation method is the same as that of Example 1, light yellow solid, yield 80%; mp 126.0-128.5℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),8.71(s,1H),7.37(d,J=8.4Hz,1H),7.14(d,J=2.4Hz,1H),6.86(dd,J1=8.4Hz,J2=2.4Hz,1H),3.7 9(s,3H),3.57(s,2H),2.83-2.71(m,2H),2.67-2.58(m,2H),2.58-2.51(m,2H),1.99-1.91(m,2H),1.58-1.45(m,4H),1.35-1.22(m,4H). 13 C NMR (100MHz, DMSO-d6) δ169.6,157.4,155.4,150.9,121.4,119.3,111.3,96.9,57.3,56.0,50.4,49.9,32.7,29.0,27.1,25.6,20.9.HRMS (ESI) m / z Calcd for C 19 H 27 N2O4(M+H) + :347.1965;Found:347.1970.

[0067] Example 6: Preparation of Compound 5f

[0068] The structural formula of compound 5f

[0069] The preparation method is the same as that of Example 1, light yellow solid, yield 64%; mp 200.5-202.3 ℃; R f =0.3 (DCM:MeOH 20:1). 1H NMR (500MHz, DMSO-d6) δ11.22(s,1H),9.06(s,1H),7.75(d,J=7.5Hz,2H),7.44(d,J=7.5Hz,2H),7.35(d,J=8.5 Hz,1H),7.16-7.06(m,1H),6.91-6.79(m,1H),3.77(s,5H),3.57(s,2H),2.83-2.70(m,2H),2.69-2.55(m,2H). 13 C NMR(125MHz,DMSO-d6)δ164.3,157.1,155.1,150.2,141.8,131.7,128.7,127.1 ,121.0,118.9,111.0,110.9,96.5,60.3,55.6,49.5,49.4,20.4.HRMS(ESI)m / z Calcd for C 20 H 21 N2O4(M+H) + :353.1496;Found:353.1490.

[0070] Example 7: Preparation of Compound 5g

[0071] The structural formula of compound 5g

[0072] The preparation method is the same as that of Example 1, light yellow solid, yield 47%; mp 153.9-156.0℃; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (400MHz, DMSO-d6) δ7.54(d,J=7.8Hz,2H),7.48-7.38(m,3H),7.36(d,J=8.4Hz,1H),7.13(d,J=2.4Hz,1H),6.85(dd,J 1=8.4,J2=2.4Hz,1H),6.47(d,J=15.8Hz,1H),3.78(s,3H),3.76(s,2H),3.58(s,2H),2.78(m,2H),2.63(d,J=5.6Hz,2H). 13C NMR(100MHz,DMSO-d6)δ163.3,157.5,155.5,150.6,140.4,138.3,134.3,129.8,128.0 ,121.4,119.4,119.3,111.40,111.3,96.9,60.8,56.0,49.9,49.8,20.8.HRMS(ESI)m / z Calcd for C 22 H 23 N2O4(M+H) + :379.1653;Found:378.1657.

[0073] Example 8: Preparation of Compound 8a

[0074] Structural formula of compound 8a

[0075] Step A: Preparation of compound 4h

[0076] In a single-necked flask, 1.62 g of compound 1a (10 mmol, 1 equiv.), 3.03 g of paraformaldehyde (100 mmol, 10 equiv.), and 2.87 g of benzylamine hydrochloride (20 mmol, 2 equiv.) were weighed and thoroughly dissolved in 20 mL of acetic acid. After stirring, the mixture was heated to 60°C and reacted for 3 hours. After TLC confirmed complete conversion, the mixture was cooled to room temperature and neutralized with saturated sodium bicarbonate solution. Ethyl acetate (3 × 100 mL) was added for extraction. The organic phases were combined, washed once with saturated brine (1 × 100 mL), and dried over anhydrous sodium sulfate. Silica gel was added and the sample was mixed. Column chromatography was performed to obtain the desired product 4h as a white solid in a 73% yield.

[0077] Step B: Preparation of compound 6

[0078] Into a double-necked empty flask, 2.13 g of 4h (7.25 mmol, 1 equiv.) and 200 mg of 10% palladium on carbon were weighed and thoroughly dissolved in 15 mL of methanol. The atmosphere was replaced with hydrogen and the reaction was stirred at room temperature for 2 hours. After TLC analysis of complete conversion, the reaction solution was filtered through celite, mixed with silica gel, and then separated by column chromatography to yield the desired product 6 as a white solid in a 91% yield.

[0079] Step C: Preparation of compound 7a

[0080] In a single-necked flask, 416 mg of 3-(4-(methoxycarbonyl)phenyl)propionic acid (2 mmol, 1 equiv.), 1.14 g of HATU (3 mmol, 1.5 equiv.), and 406 mg of compound 6 (2 mmol, 1 equiv.) were weighed and thoroughly dissolved in 6 mL of DMF. 0.7 mL of DIPEA (4 mmol, 2 equiv.) was added and stirred at room temperature for 2 hours. The reaction was incubated at 60°C for 6 hours. After complete conversion as determined by TLC, 20 mL of water was added and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, washed with saturated brine (5 × 20 mL), and then dried over anhydrous sodium sulfate. Silica gel was added and the sample was mixed. The desired product 7a was isolated by column chromatography as a white solid in a 73% yield.

[0081] Step D: Preparation of compound 8a

[0082] 96 mg of lithium hydroxide (4 mmol, 8 equiv.) and hydroxylamine aqueous solution (5 mmol, 10 equiv.) were weighed into a single-necked flask and dissolved in 2 mL of methanol. At -10°C, 197 mg of compound 7a (0.5 mmol, 1 equiv.) was dissolved in 3 mL of methanol and slowly added dropwise to the reaction solution. After stirring at -10°C for 30 minutes, the mixture was slowly warmed to room temperature and stirred for 8 hours. The reaction solution was then dried, 10 mL of water was added, and the pH was adjusted to 3-4 with 2M hydrochloric acid. The mixture was filtered, and the residue was washed twice with water and once with ethyl acetate. The filter cake was collected and dried to obtain compound 8a as a white solid with a yield of 55%; mp 198.1-199.5°C; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (400MHz, DMSO-d6) δ11.17(s,1H),9.00(s,1H),7.73-7.62(m,2H),7.43-7.30(m,3H),7.17(d,J=2.2Hz,1H), 6.87(dd,J1=8.4Hz,J2=2.2Hz,1H),4.70-4.60(m,2H),3.86-3.64(m,5H),2.95-2.75(m,4H),2.72-2.56(m,2H). 13 CNMR(100MHz,DMSO-d6)δ171.3,164.7,157.7,155.5,149.0,145.4,130.9,128.9,127.3, 121.1,119.6,112.4,111.6,96.9,56.1,43.0,34.6,34.1,30.9,21.5,20.6.HRMS(ESI)m / z Calcd for C22 H 23 N2O5(M+H) + :395.4345;Found:395.4345.

[0083] According to the preparation method of Example 8, appropriate starting materials were selected to prepare the compounds of Examples 9 and 10.

[0084] Example 9: Preparation of Compound 8b

[0085] Structural formula of compound 8b

[0086] The preparation method is the same as that of Example 8, white solid, yield 53%; mp218.6-220.4℃; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (400MHz, DMSO-d6) δ11.33(s,1H),9.14(s,1H),7.94-7.75(m,4H),7.64-7.35(m,3H),7.21(d,J=2.2Hz,1H),6.90( dd, J1=8.4Hz, J2=2.2Hz,1H),5.06-4.70(m,2H),4.17-3.87(m,2H),3.81(s,3H),3.43-3.15(m,1H),2.84-2.64(m,2H). 13 C NMR(100MHz,DMSO-d6)δ165.9,164.1,157.8,155.6,148.9,141.6,138.2,133.8,128.7 ,127.7,121.1,120.4,119.6,112.0,111.7,96.9,56.1,43.4,41.0,22.0.HRMS(ESI)m / z Calcd for C 22 H 21 N2O5(M+H) + : 393.1445; Found:393.1447.

[0087] Example 10: Preparation of Compound 8c

[0088] Structural formula of compound 8c

[0089] The preparation method is the same as that of Example 8, white solid, yield 57%; mp 171.5-173.5℃; R f =0.3 (DCM:MeOH 10:1). 1H NMR (400MHz, DMSO-d6) δ11.07(s,1H),8.92(s,1H),7.70(d,J=8.4Hz,2H),7.42(d,J=8.4Hz,1H),7.19(s,1H),7.0 5-6.95(m,2H),6.88(d,J=8.4Hz,1H),5.18-4.98(m,2H),4.76-4.57(m,2H),3.87-3.66(m,5H),2.85-2.60(m,2H). 13 C NMR(100MHz,DMSO-d6)δ167.1,164.5,160.8,157.8,155.7,155.6,148.5,148.2,129.0,125.8,1 21.0,119.7,114.8,112.4,112.0,111.7,96.9,66.4,56.1,42.5,42.3,21.5,20.5.HRMS(ESI)m / z Calcd for C 21 H 21 N2O6(M+H) + :397.1394; Found:397.1392.

[0090] Example 11: Preparation of Compound 8d

[0091] The structural formula of compound 8d

[0092] Step A: Preparation of compound 4i

[0093] In a single-necked flask, 1.62 g of 1a (10 mmol, 1 equiv.), 3.03 g of paraformaldehyde (100 mmol, 10 equiv.), and 2.32 g of chloroethylamine hydrochloride (20 mmol, 2 equiv.) were weighed and thoroughly dissolved in 20 mL of acetic acid. After stirring, the mixture was heated to 60°C and reacted for 3 hours. After TLC confirmed complete conversion, the mixture was cooled to room temperature and neutralized with saturated sodium bicarbonate solution. Ethyl acetate (3 × 100 mL) was added for extraction. The organic phases were combined, washed once with saturated brine (1 × 100 mL), and dried over anhydrous sodium sulfate. Silica gel was added, the sample was mixed, and column chromatography was performed to obtain the desired product 4i as a white solid in a 76% yield.

[0094] Step B: Preparation of compound 7d

[0095] In a single-necked flask, 2.01 g of compound 4i (7.56 mmol, 1 equiv.), 1.15 g of methyl parahydroxybenzoate (7.56 mmol, 1 equiv.), 2.09 g of potassium carbonate (15.1 mmol, 2 equiv.), and 249 mg of potassium iodide (1.5 mmol, 0.2 equiv.) were weighed and thoroughly dissolved in 30 mL of acetonitrile. The mixture was reacted at 80°C for 3 hours. After complete conversion by TLC, 60 mL of water was added and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, washed once with saturated brine (1 × 50 mL), and dried over anhydrous sodium sulfate. Silica gel was added and the sample was mixed. The desired product 7d was isolated by column chromatography as a white solid in a 77% yield.

[0096] Step C: Preparation of compound 8d

[0097] 96 mg of lithium hydroxide (4 mmol, 8 equiv.) and hydroxylamine aqueous solution (5 mmol, 10 equiv.) were weighed into a single-necked flask and dissolved in 2 mL of methanol. At -10°C, 191 mg of compound 7d (0.5 mmol, 1 equiv.) was dissolved in 3 mL of methanol and slowly added dropwise to the reaction solution. After stirring at -10°C for 30 minutes, the mixture was slowly warmed to room temperature and stirred for 8 hours. The reaction solution was then dried and 10 mL of water was added. The pH was then adjusted to 3-4 with 2 M hydrochloric acid. The mixture was filtered, and the residue was washed twice with water and once with ethyl acetate. The filter cake was collected and dried to obtain compound 8d as a white solid with a yield of 65%; mp 145.0-146.8°C; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),8.95(s,1H),7.75(d,J=8.4Hz,2H),7.38(d,J=8.4Hz,1H),7.16(s,1H), 7.04(d,J=8.4Hz,2H),6.87(d,J=8.4Hz,1H),4.25(s,2H),3.95-3.61(m,5H),3.13-2.81(m,4H),2.66(s,2H). 13 C NMR(100MHz,DMSO-d6)δ164.5,161.2,157.5,155.5,150.6,129.2,125.5,121.4,1 19.4,114.7,111.4,111.3,96.9,66.5,56.0,55.8,50.6,50.2,20.8.HRMS(ESI)m / z Calcd for C 21 H 23N2O5(M+H) + :383.1601;Found:383.1602.

[0098] Example 12: Preparation of Compound 12a

[0099] Structural formula of compound 12a

[0100] Step A: Synthesis of compound 4a

[0101] In a single-necked flask, 1.62 g of compound 1a (10 mmol, 1 equiv.), 3.03 g of paraformaldehyde (100 mmol, 10 equiv.), and 2.51 g of glycine methyl ester hydrochloride (20 mmol, 2 equiv.) were weighed and thoroughly dissolved in 20 mL of acetic acid. After stirring, the mixture was heated to 60°C and reacted for 3 hours. After TLC confirmed complete conversion, the mixture was cooled to room temperature and neutralized with saturated sodium bicarbonate solution. Ethyl acetate (3 × 100 mL) was then added for extraction. The organic phases were combined, washed once with saturated brine (1 × 100 mL), and dried over anhydrous sodium sulfate. Silica gel was added and the sample was mixed. Column chromatography was performed to obtain the desired product 4a as a white solid in a 76% yield.

[0102] Step B: Synthesis of compound 10a

[0103] In a single-necked flask, 550 mg of 4a (2 mmol, 1 equiv.) was weighed and dissolved in 4 mL of MeOH and 2 mL of water. 96 mg of lithium hydroxide (4 mmol, 2 equiv.) was added and allowed to react at room temperature for 8 h. The reaction solution was spin-dried, 10 mL of water was added, and the pH was adjusted to 1-2 with 4 M hydrochloric acid. The reaction solution was filtered, and the filter cake was washed with water and petroleum ether to obtain compound 10a as a white solid in 80% yield.

[0104] Step C: Preparation of compound 11a

[0105] In a single-necked flask, 261 mg of compound 10a (1 mmol, 1 equiv.), 570 mg of HATU (1.5 mmol, 1.5 equiv.), and 151 mg of methyl p-aminobenzoate (1 mmol, 1 equiv.) were weighed and thoroughly dissolved in 6 mL of DMF. 0.35 mL of DIPEA (2 mmol, 2 equiv.) was added and stirred at room temperature for 2 hours. The mixture was then heated to 60°C and reacted for 6 hours. After TLC confirmed complete conversion, 20 mL of water was added, and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, washed with saturated brine (5 × 20 mL), and dried over anhydrous sodium sulfate. Silica gel was added and the sample was mixed. Column chromatography was then performed to obtain the desired product 11a as a pale yellow oil in a 67% yield.

[0106] Step D: Preparation of compound 12a

[0107] 96 mg of lithium hydroxide (4 mmol, 8 equiv.) and hydroxylamine aqueous solution (5 mmol, 10 equiv.) were weighed into a single-necked flask and dissolved in 2 mL of methanol. At -10°C, 197 mg of compound 11a (0.5 mmol, 1 equiv.) was dissolved in 3 mL of methanol and slowly added dropwise to the reaction solution. After stirring at -10°C for 30 minutes, the mixture was slowly warmed to room temperature and stirred for 8 hours. The reaction solution was then dried, 10 mL of water was added, and the pH was adjusted to 3-4 with 2M hydrochloric acid. The mixture was filtered, and the residue was washed twice with water and once with ethyl acetate. The filter cake was collected and dried to obtain compound 12a as a white solid with a yield of 84%; mp 199.4-201.5°C; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (500MHz, DMSO-d6) δ11.12(s,1H),10.05(s,1H),8.96(s,1H),7.76-7.68(m,4H),7.39(d,J=8.5Hz,1H),7.15(d,J=2.0Hz ,1H),6.86(dd,J1=8.5Hz,J2=2.0Hz,1H),3.84-3.79(m,2H),3.79(s,3H),3.45(s,2H),2.96-2.89(m,2H),2.73-2.65(m,2H). 13C NMR (125MHz, DMSO-d6) δ169.4,157.5,155.4,150.3,141.7,128.1,127.9,121.4 ,119.3,119.3,111.4,111.2,96.9,60.7,56.0,50.1,49.7,20.6.HRMS(ESI)m / z Calcd for C 21 H 22 N3O5(M+H) + :396.1554;Found:396.1550.

[0108] According to the preparation method of Example 12, appropriate starting materials were selected to prepare the compounds of Examples 13 to 38.

[0109] Example 13: Preparation of Compound 12b

[0110] Structural formula of compound 12b

[0111] The preparation method is the same as that of Example 12, light yellow solid, yield 69%; mp 201.1-203.2℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(600MHz,DMSO-d6)δ11.15(s,1H),10.15(s,1H),8.96(s,1H),7.74(s,4H),7.28-7.03(m ,2H),6.76(d,J=7.8Hz,1H),3.86(s,5H),3.50(s,2H),3.05-2.91(m,2H),2.91-2.74(m,2H). 13 C NMR(150MHz,DMSO-d6)δ169.1,164.4,155.6,154.2,149.5,141.7,128.1,127.9,1 24.9,119.3,117.4,110.7,104.8,104.4,60.5,56.0,50.2,49.7,22.3.HRMS(ESI) m / z Calcd for C 21 H 22 N3O5(M+H) + :396.1554;Found:396.1550.

[0112] Example 14: Preparation of Compound 12c

[0113] Structural formula of compound 12c

[0114] The preparation method is the same as that of Example 12, light yellow solid, yield 66%; mp 195.5-197.3 ℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(500MHz,DMSO-d6)δ10.08(s,1H),7.73(s,4H),7.21(s,1H),7.05(s,1 H),3.85-3.74(m,8H),3.45(s,2H),2.98-2.88(m,2H),2.75–2.63(m,2H). 13 C NMR(125MHz,DMSO-d6)δ169.4,164.2,150.1,148.9,147.4,146.6,141.6,128.0,1 19.9,119.3,111.6,101.7,96.7,60.7,56.5,56.4,50.2,49.9,20.8.HRMS(ESI)m / z Calcd for C 22 H 24 N3O6(M+H) + :426.1660; Found:426.1654.

[0115] Example 15: Preparation of Compound 12d

[0116] Structural formula of compound 12d

[0117] The preparation method is the same as that of Example 12, white solid, yield 76%; mp 229.2-231.4℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(500MHz,DMSO-d6)δ11.15(s,1H),10.08(s,1H),8.98(s,1H),7.74(s,4H),7.21(s,1 H),7.01(s,1H),6.03(s,2H),3.80(s,2H),3.45(s,2H),3.05-2.85(m,2H),2.66(s,2H). 13 C NMR (125MHz, DMSO-d6) δ169.3,164.4,150.6,149.3,145.4,144.4,141.7,128.1,12 7.9,121.4,119.3,111.9,101.6,98.3,94.3,60.6,50.1,49.8,20.6.HRMS(ESI)m / z Calcd for C 21 H20 N3O6(M+H) + :410.1347; Found:410.1358.

[0118] Example 16: Preparation of Compound 12e

[0119] Structural formula of compound 12e

[0120] The preparation method is the same as that of Example 12, light yellow solid, yield 83%; mp 184.9-188.3℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR (400MHz, DMSO-d6) δ11.16(s,1H),10.09(s,1H),9.00(s,1H),7.75(s,4H),7.13(d,J=8.4Hz,1H),7. 00(d,J=8.4Hz,1H),3.95(s,3H),3.90–3.80(m,5H),3.48(s,2H),3.01-2.89(m,2H),2.76-2.65(m,2H). 13 C NMR(100MHz,DMSO-d6)δ169.4,164.4,150.9,149.1,146.7,141.7,134.7,128.1,127.9,1 24.0,119.3,112.8,111.5,110.2,60.9,60.6,57.4,55.4,49.9,49.8,20.6.HRMS(ESI)m / z Calcd for C 22 H 24 N3O6(M+H) + :426.1660;Found:426.1664.

[0121] Example 17: Preparation of Compound 12f

[0122] The structural formula of compound 12f

[0123] The preparation method is the same as that of Example 12, light yellow solid, yield 48%; mp 247.2-248.0℃; R f =0.3 (DCM:MeOH 20:1). 1H NMR (400MHz, DMSO-d6) δ11.18(s,1H),10.06(s,1H),9.46(s,1H),8.99(s,1H),7.74(s,4H),7.28(d,J=8.4H z,1H),6.89(s,1H),6.73(d,J=8.4Hz,1H),3.80(s,2H),3.45(s,2H),3.09-2.82(m,2H),2.77-2.58(m,2H). 13 C NMR(100MHz,DMSO-d6)δ169.4,155.6,155.4,149.5,141.7,128.0,127.8,120.2 ,119.3,119.3,111.9,111.2,98.4,60.7,55.4,50.1,49.8,20.7.HRMS(ESI)m / z Calcd for C 20 H 20 N3O5(M+H) + :382.1397;Found:382.1397.

[0124] Example 18: Preparation of Compound 12g

[0125] The structural formula of compound 12g

[0126] The preparation method is the same as that of Example 12, light yellow solid, yield 72%; mp 201.2-204.6 ℃; R f =0.3 (DCM: MeOH 20:1). 1 H NMR(500MHz,DMSO-d6)δ10.06(s,1H),7.73(s,4H),7.47-7.32(m,2H),3.8 6(s,3H),3.82(s,2H),3.46(s,2H),2.99-2.85(m,2H),2.73-2.60(m,2H). 13 C NMR(125MHz,DMSO-d6)δ169.3,151.5,150.6,150.5,148.6,145.3,145.2,141.6,128.0,127.9,12 0.1,119.9,119.3,111.7,111.6,105.6,105.4,97.9,60.6,56.9,49.9,49.7,20.6.HRMS(ESI)m / z Calcd for C 21 H 21 FN3O5(M+H) +:414.1460; Found:414.1468.

[0127] Example 19: Preparation of Compound 12h

[0128] The structural formula of compound 12h

[0129] The preparation method is the same as that of Example 12, light yellow solid, yield 67%; mp 217.9-218.9 ℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(600MHz,DMSO-d6)δ11.12(s,1H),10.05(s,1H),8.95(s,1H),7.73(s,4H),7.59(s,1H) ,7.41(s,1H),3.88(s,3H),3.83(s,2H),3.46(s,2H),2.98-2.86(m,2H),2.78-2.62(m,2H). 13 C NMR(150MHz,DMSO-d6)δ169.3,164.4,153.8,152.2,151.5,141.7,128.1,127.9,12 1.6,119.6,119.3,117.3,111.1,97.2,60.6,56.9,49.9,49.6,20.6.HRMS(ESI)m / z Calcd for C 21 H 21 ClN3O5(M+H) + :430.1164; Found:430.1170.

[0130] Example 20: Preparation of Compound 12i

[0131] Structural formula of compound 12i

[0132] The preparation method is the same as that of Example 12, light yellow solid, yield 61%; mp 207.0-209.2℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(400MHz,DMSO-d6)δ11.15(s,1H),10.08(s,1H),8.99(s,1H),7.74(s,4H),7.26(s,1H) ,7.16(s,1H),3.82(s,5H),3.46(s,2H),2.99-2.87(m,2H),2.77-2.62(m,2H),2.23(s,3H). 13C NMR(100MHz,DMSO-d6)δ169.4,164.4,155.5,154.0,149.7,141.7,128.1,127.9,121.5,1 20.4,119.8,119.3,111.1,94.8,60.8,56.1,55.4,50.2,49.9,20.7,16.9.HRMS(ESI)m / z Calcd for C 22 H 24 N3O5(M+H) + :410.1710;Found:410.1715.

[0133] Example 21: Preparation of Compound 12j

[0134] Structural formula of compound 12j

[0135] The preparation method is the same as that of Example 12, light yellow solid, yield 56%; mp 189.9-192.0℃; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (500MHz, DMSO-d6) δ10.14(s,1H),7.74(s,4H),7.34(d,J=8.5Hz,1H),7.14(d,J=2.5Hz,1H),6.84(dd,J1=8.5Hz,J2=2.5H z,1H),3.92-3.84(m,1H),3.82-3.73(m,4H),3.64-3.54(m,1H),2.90-2.81(m,2H),2.68-2.59(m,2H),1.33(d,J=7.0Hz,3H). 13 C NMR (125MHz, DMSO-d6) δ172.3,164.3,157.4,155.5,150.7,141.7,128.0,127.9,121. 4,119.3,111.4,111.3,96.8,63.2,55.9,47.1,46.5,25.9,21.4,13.3.HRMS(ESI)m / z Calcd for C 22 H 24 N3O5(M+H) + :410.1710;Found:410.1714.

[0136] Example 22: Preparation of Compound 12k

[0137] Structural formula of compound 12k

[0138] The preparation method is the same as that of Example 12, light yellow solid, yield 58%; mp 168.8-171.4℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(500MHz,DMSO-d6)δ11.16(s,1H),10.20(s,1H),9.01(s,1H),7.74(d,J=8.5Hz,2H),7.67(d,J=8.5Hz,2H),7.43-7.28(m,5H),7 .24-7.15(m,2H),6.96-6.82(m,1H),4.04-3.90(m,2H),3.89-3.77(m,4H),3.33-3.20(m,1H),3.14-3.00(m,3H),2.71-2.61(m,2H). 13 C NMR (125MHz, DMSO-d6) δ170.1,164.3,157.4,155.4,150.7,141.5,139.2,129.6,128.7,128.1,1 27.9,126.6,121.4,119.3,119.2,111.3,96.8,69.5,56.0,47.2,46.6,34.4,21.6.HRMS(ESI)m / z Calcd for C 28 H 28 N3O5(M+H) + :486.2023;Found:486.2024.

[0139] Example 23: Preparation of Compound 121

[0140] Structural formula of compound 12l

[0141] The preparation method is the same as that of Example 12, light yellow solid, yield 45%; mp 123.1-126.5℃; R f =0.3 (DCM:MeOH 20:1). 1H NMR (500MHz, DMSO-d6) δ11.14(s,1H),10.29(s,1H),8.98(s,1H),7.72(s,4H),7.34(d,J=8.5Hz,1H),7.13(d,J=2.5Hz,1H),6.84(dd,J1= 8.5Hz, J2=2.5Hz,1H),3.96-3.75(m,5H),3.70-3.62(m,1H),2.99-2.89(m,2H),2.65-2.58(m,2H),2.59-2.51(m,2H),2.14-1.98(m,5H). 13 C NMR(125MHz,DMSO-d6)δ170.2,163.9,157.0,155.0,150.3,141.2,127.7,127.5,120.9,118 .9,118.8,110.9,110.9,96.4,65.9,55.6,46.6,46.0,30.3,27.3,21.2,14.8.HRMS(ESI)m / z Calcd for C 24 H 28 N3O5S(M+H) + :470.1744;Found:470.1740.

[0142] Example 24: Preparation of Compound 12m

[0143] The structural formula of compound 12m

[0144] The preparation method is the same as that of Example 12, white solid, yield 40%; mp 134.0-139.5℃; R f =0.2 (DCM:MeOH 20:1). 1H NMR (500MHz, DMSO-d6) δ10.13(s,1H),9.18(s,1H),7.70(d,J=8.5Hz,2H),7.63(d,J=8.5Hz, 2H),7.34(d,J=8.5Hz,1H),7.13(d,J=2.5Hz,1H),7.07(d,J=8.0Hz,2H),6.84(dd,J1=8.5Hz, J2=2.5Hz,1H),6.65(d,J=8.0Hz,2H),3.99-3.92(m,1H),3.91-3.82(m,1H),3.78(s,3H),3. 76-3.69(m,1H),3.16-3.06(m,1H),3.03-2.96(m,2H),2.95-2.87(m,1H),2.67-2.54(m,2H). 13 C NMR (125MHz, DMSO-d6) δ170.4,169.9,163.8,156.9,155.7,155.0,150.4,141.1,130.1,128.5,127. 7,127.5,120.9,118.8,118.8,115.1,110.9,96.4,69.4,55.6,46.9,46.1,33.3,21.1.HRMS(ESI)m / z Calcd for C 28 H 28 N3O6(M+H) + :502.1973;Found: 502.1979.

[0145] Example 25: Preparation of Compound 12n

[0146] The structural formula of compound 12n

[0147] The preparation method is the same as that of Example 12, light yellow solid, yield 35%; mp 188.5-187.4℃; R f =0.3 (DCM:MeOH 10:1). 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H),10.13(s,1H),9.09-8.75(m,1H),7.68(d,J=8.4Hz,2H),7.60(d,J=8.4Hz,2H),7.29-7.24(m,4H) ,7.19-7.14(m,2H),6.97(s,1H),6.00(s,2H),3.99-3.85(m,2H),3.84-3.64(m,1H),3.23-3.17(m,1H),3.04-2.97(m,3H),2.56(s,2H). 13 C NMR (100MHz, DMSO-d6) δ170.1,164.4,151.2,149.3,145.3,144.4,141.6,139.2,129.7,128.7,128. 2,128.0,126.7,121.4,119.2,112.0,101.6,98.3,94.3,69.5,47.2,46.7,34.4,21.6.HRMS(ESI)m / z Calcd for C 28 H 26 N3O6(M+H) + :500.1816;Found:500.1817.

[0148] Example 26: Preparation of Compound 12o

[0149] The structural formula of compound 12o

[0150] The preparation method is the same as that of Example 12, light yellow solid, yield 41%; mp 170.5-172.3 ℃; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),7.71(s,4H),7.34(d,J=8.4Hz,1H),7.14(d,J=2.2Hz,1H), 6.84(dd,J1=8.4,J2=2.2Hz,1H),4.02-3.98(m,1H),3.84(s,2H),3.78(s,3H),3.02–2.55(m,6H). 13C NMR (100MHz, CD3OD) δ174.41,170.53,164.82,157.97,155.97,151.10,142.34,128.66,128.30,121. 89,119.82,119.63,111.87,111.83,97.36,64.92,56.52,47.39,47.21,32.70,22.15.HRMS(ESI)m / z Calcd for C 23 H 24 N3O7(M+H) + : 454.1609; Found:454.1621.

[0151] Example 27: Preparation of Compound 12p

[0152] Structural formula of compound 12p

[0153] The preparation method is the same as that of Example 12, light yellow solid, yield 34%; mp 171.6-173.4 ℃; R f =0.3 (DCM:MeOH 10:1). 1 H NMR(400MHz,CD3OD)δ7.66(d,J=7.8Hz,2H),7.51(d,J=8.2Hz,2H),7.33-7.15(m, 7H),4.38-4.12(m,2H),3.96(s,1H),3.89(s,3H),3.43-3.29(m,4H),2.84(s,2H). 13 C NMR (100MHz, CD3OD) δ167.6,152.6,152.5,150.2,149.9,147.3,142.0,137.8and137.7,134.1and 134.0,130.5,129.7,129.2,128.9,128.1,120.8,112.8,106.0,105.8,98.2,71.2,57.1,48.14,21.2.HRMS(ESI)m / z Calcd for C 28 H 27 FN3O5(M+H) + :504.1929;Found:504.1930.

[0154] Example 28: Preparation of Compound 12q

[0155] The structural formula of compound 12q

[0156] The preparation method was the same as that of Example 12, and the product was a white solid with a yield of 59%, mp 168.8–171.4°C; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(500MHz,DMSO-d6)δ11.16(s,1H),10.20(s,1H),9.01(s,1H),7.74(d,J=8.5Hz,2H),7.67(d,J=8.5Hz,2H),7.43-7.28(m,5H),7 .24-7.15(m,2H),6.96-6.82(m,1H),4.04-3.90(m,2H),3.89-3.77(m,4H),3.33-3.20(m,1H),3.14-3.00(m,3H),2.71-2.61(m,2H). 13 C NMR (125MHz, DMSO-d6) δ170.1,164.3,157.4,155.4,150.7,141.5,139.2,129.6,128.7,128.1,1 27.9,126.6,121.4,119.3,119.2,111.3,96.8,69.5,56.0,47.2,46.6,34.4,21.6.HRMS(ESI)m / z Calcd for C 28 H 28 N3O5(M+H) + :486.2023;Found:486.2023.

[0157] Example 29: Preparation of Compound 12r

[0158] The structural formula of compound 12r

[0159] The preparation method is the same as that of Example 12, white solid, yield 43%; mp 134.1-139.3 ℃; R f =0.2 (DCM:MeOH 20:1). 1H NMR (500MHz, DMSO-d6) δ10.13(s,1H),9.18(s,1H),7.70(d,J=8.5Hz,2H),7.63(d,J=8.5Hz, 2H),7.34(d,J=8.5Hz,1H),7.13(d,J=2.5Hz,1H),7.07(d,J=8.0Hz,2H),6.84(dd,J1=8.5Hz, J2=2.5Hz,1H),6.65(d,J=8.0Hz,2H),3.99-3.92(m,1H),3.91-3.82(m,1H),3.78(s,3H),3. 76-3.69(m,1H),3.16-3.06(m,1H),3.03-2.96(m,2H),2.95-2.87(m,1H),2.67-2.54(m,2H). 13 C NMR (125MHz, DMSO-d6) δ170.4,169.9,163.8,156.9,155.7,155.0,150.4,141.1,130.1,128.5,127. 7,127.5,120.9,118.8,118.8,115.1,110.9,96.4,69.4,55.6,46.9,46.1,33.3,21.1.HRMS(ESI)m / z Calcd for C 28 H 28 N3O6(M+H) + :502.1973;Found:502.1979.

[0160] Example 30: Preparation of Compound 12s

[0161] Structural formula of compound 12s

[0162] The preparation method is the same as that of Example 12, light yellow solid, yield 56%; mp 189.9-192.0℃; R f =0.3 (DCM:MeOH 10:1). 1H NMR (400MHz, DMSO-d6) δ11.13(s,1H),10.11(s,1H),8.96(s,1H),7.81–7.67(m,4H),7.38(d,J=8.4Hz,1H),7.16(d,J=2.2Hz,1H),6.87( dd,J1=8.4,J2=2.2Hz,1H),3.95–3.72(m,5H),3.61(q,J=6.6Hz,1H),2.87(d,J=5.4Hz,2H),2.67(d,J=5.4Hz,2H),1.34(d,J=6.6Hz,3H). 13 C NMR (100MHz, DMSO-d6) δ172.3,164.4,157.5,155.5,150.8,141.8,128.1,127. 9,121.4,119.3,111.5,111.4,96.9,63.2,56.0,47.1,46.5,26.0,21.4,13.4. HRMS(ESI)m / z Calcd for C 22 H 24 N3O5(M+H) + :410.1710;Found:410.1714.

[0163] Example 31: Preparation of Compound 12t

[0164] Structural formula of compound 12t

[0165] The preparation method is the same as that of Example 12, white solid, yield 42%; mp 191.2-194.0℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR(400MHz,DMSO-d6)δ11.07(s,1H),10.19(s,1H),8.90(s,1H),7.71–7.61(m,4H) ,7.29(d,J=8.4Hz,1H),7.08(d,J=2.2Hz,1H),6.79(dd,J1=8.4,J2=2.2Hz,1H),3.9 1–3.83(m,1H),3.77–3.68(m,4H),3.57–3.50(m,1H),2.88(t,J=5.6Hz,2H),2.58–2 .49(m,2H),1.73–1.61(m,1H),1.60–1.50(m,2H),0.89(dd,J1=14.8,J2=5.8Hz,6H). 13C NMR(100MHz,DMSO-d6)δ171.7,164.9,157.9,156.0,151.4,142.2,128.7,128.5, 122.0,119.8,111.9,97.4,66.3,56.5,47.8,46.9,38.1,25.8,23.8,23.5,22.2.

[0166] Example 32: Preparation of Compound 12u

[0167] Structural formula of compound 12u

[0168] The preparation method is the same as that of Example 12, white solid, yield 44%; f =0.3 (DCM:MeOH 20:1). 1 H NMR (400MHz, DMSO-d6) δ11.08(s,1H),10.37(s,1H),8.91(s,1H),7.67(s,4H),7.58–7.50(m,2H),7.40–7.28(m,4H),7.07(d,J=2. 2Hz,1H),6.82(dd,J1=8.4,J1=2.2Hz,1H),4.39(s,1H),3.73(s,3H),3.67–3.54(m,2H),2.90–2.74(m,2H),2.61(d,J=5.6Hz,2H). 13 C NMR(100MHz,DMSO-d6)δ170.1,164.3,157.6,155.5,150.2,141.7,137.5,129.2,12 9.1,128.8,128.2,121.4,119.5,119.4,111.5,96.9,73.3,56.1,48.0,47.8,20.6.

[0169] Example 33: Preparation of Compound 12v

[0170] Structural formula of compound 12v

[0171] The preparation method is the same as that of Example 12, white solid, yield 72%; R f =0.3 (DCM:MeOH 10:1). 1H NMR(400MHz,DMSO-d6)δ11.13(s,1H),10.16(s,1H),8.97(s,1H),7.74(s,4H), 7.37(d,J=8.4Hz,1H),7.15(d,J=2.2Hz,1H),6.86(dd,J1=8.4,J2=2.2Hz,1H), 4.92(t,J=5.2Hz,1H),4.09–4.00(m,1H),4.01–3.90(m,1H),3.88–3.81(m,2H) ,3.80(s,3H),3.56(t,J=6.0Hz,1H),3.05–2.91(m,2H),2.65(d,J=5.2Hz,2H). 13 C NMR(100MHz,DMSO-d6)δ170.6,164.4,157.5,155.5,150.8,141.7,128.1,1 27.9,121.5,119.3,119.2,111.4,96.9,69.8,60.1,56.0,48.0,46.9,21.5.

[0172] Example 34: Preparation of Compound 12w

[0173] Structural formula of compound 12w

[0174] The preparation method is the same as that of Example 12, light yellow solid, yield 41%; mp 170.5-172.3 ℃; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),7.71(s,4H),7.34(d,J=8.4Hz,1H),7.14(d,J=2.2Hz,1H), 6.84(dd,J1=8.4,J2=2.2Hz,1H),4.02-3.98(m,1H),3.84(s,2H),3.78(s,3H),3.02–2.55(m,6H). 13 C NMR (100MHz, CD3OD) δ174.41,170.53,164.82,157.97,155.97,151.10,142.34,128.66,128.30,121. 89,119.82,119.63,111.87,111.83,97.36,64.92,56.52,47.39,47.21,32.70,22.15.HRMS(ESI)m / z Calcd for C 23 H 24N3O7(M+H) + :454.1609; Found:454.1621.

[0175] Example 35: Preparation of Compound 12x

[0176] Structural formula of compound 12x

[0177] The preparation method is the same as that of Example 12, yellow solid, yield 45%; mp 123.1-126.5℃; R f =0.3 (DCM:MeOH 20:1). 1 H NMR (500MHz, DMSO-d6) δ11.14(s,1H),10.29(s,1H),8.98(s,1H),7.72(s,4H),7.34(d,J=8.5Hz,1H),7.13(d,J=2.5Hz,1H),6.84(dd,J1= 8.5Hz, J2=2.5Hz,1H),3.96-3.75(m,5H),3.70-3.62(m,1H),2.99-2.89(m,2H),2.65-2.58(m,2H),2.59-2.51(m,2H),2.14-1.98(m,5H). 13 C NMR(125MHz,DMSO-d6)δ170.2,163.9,157.0,155.0,150.3,141.2,127.7,127.5,120.9,118 .9,118.8,110.9,110.9,96.4,65.9,55.6,46.6,46.0,30.3,27.3,21.2,14.8.HRMS(ESI)m / z Calcd for C 24 H 28 N3O5S(M+H) + :470.1744;Found:470.1740.

[0178] Example 36: Preparation of Compound 12y

[0179] The structural formula of compound 12y

[0180] The preparation method is the same as that of Example 12, white solid, yield 29%; R f =0.3 (DCM:MeOH 10:1). 1H NMR (400MHz, DMSO-d6) δ11.08(s,1H),10.12(s,1H),8.93(s,1H),7.69(d,J=8.4Hz,2H),7.62(d,J=8.4Hz,2H),7.33–7.22(m,4H) ,7.21–7.12(m,2H),7.00(s,1H),3.98–3.83(m,2H),3.82–3.72(m,7H),3.27–3.16(m,1H),3.08–2.94(m,3H),2.66–2.56(m,2H). 13 C NMR(100MHz,DMSO-d6)δ170.2,164.4,150.6,148.9,147.4,146.6,141.6,139.2,129.7,128.7,1 28.2,128.0,126.7,120.0,119.3,111.7,101.8,96.7,69.6,56.5,56.5,47.3,46.8,34.4,21.8.

[0181] Example 37: Preparation of Compound 12z

[0182] The structural formula of compound 12z

[0183] The preparation method is the same as that of Example 12, light yellow solid, yield 35%; mp 188.5-187.4℃; R f =0.3 (DCM:MeOH 10:1). 1 H NMR (400MHz, DMSO-d6) δ11.09(s,1H),10.13(s,1H),9.09-8.75(m,1H),7.68(d,J=8.4Hz,2H),7.60(d,J=8.4Hz,2H),7.29-7.24(m,4H) ,7.19-7.14(m,2H),6.97(s,1H),6.00(s,2H),3.99-3.85(m,2H),3.84-3.64(m,1H),3.23-3.17(m,1H),3.04-2.97(m,3H),2.56(s,2H). 13C NMR (100MHz, DMSO-d6) δ170.1,164.4,151.2,149.3,145.3,144.4,141.6,139.2,129.7,128.7,128. 2,128.0,126.7,121.4,119.2,112.0,101.6,98.3,94.3,69.5,47.2,46.7,34.4,21.6.HRMS(ESI)m / z Calcd for C 28 H 26 N3O6(M+H) + :500.1816;Found:500.1817.

[0184] Example 38: Preparation of Compound 12aa

[0185] The structural formula of compound 12aa

[0186] The preparation method is the same as that of Example 12, white solid, yield 47%; R f =0.3 (DCM:MeOH 10:1). 1 H NMR(400MHz, DMSO-d6)δ11.06(s,1H),10.10(s,1H),8.90(d,J=2.0Hz,1H),7.69–7.63(m,2H),7.62–7.56(m,2H),7.52(s,1H),7.36(s,1H),7.29 –7.21(m,4H),7.18–7.10(m,1H),3.99–3.87(m,2H),3.85(s,3H),3.81–3 .74(m,1H),3.23–3.14(m,1H),3.06–2.92(m,3H),2.58(d,J=4.6Hz,2H). 13 C NMR (100MHz, DMSO-d6) δ170.1,153.9,152.2,152.0,141.5,139.2,129.7,128.7,128.2,128. 0,126.7,121.7,119.6,119.3,117.3,111.3,97.3,69.5,57.0,47.2,46.6,34.4,29.5,21.5.

[0187] Example 39: Preparation of Compound 12ab

[0188] The structural formula of compound 12ab

[0189] Step A: Preparation of compound 13

[0190] In a single-necked flask, 1.14 g of 11ab (3 mmol, 1 equiv.) was weighed and dissolved in 10 mL of anhydrous DCM. 0.5 mL of pyridine (6 mmol, 2 equiv.) and 1.0 mL of trifluoromethanesulfonic anhydride (6 mmol, 2 equiv.) were added at -10°C and allowed to warm to room temperature for 2 hours. After complete conversion as determined by TLC, 2 mL of 2M hydrochloric acid was added to the reaction mixture at 0°C. The mixture was extracted with DCM (3 × 20 mL). The organic phases were combined, washed with saturated sodium bicarbonate solution and saturated brine, respectively, and dried over anhydrous sodium sulfate. Silica gel was added to the sample, and the mixture was separated by column chromatography to obtain the intermediate.

[0191] Under anhydrous and oxygen-free conditions, the intermediate obtained in the previous step was dissolved in 6 mL of DMF, and 0.6 mL of triethylamine (4 mmol, 2 equiv.), 0.4 mL of formic acid (10 mmol, 5 equiv.), and 45 mg of palladium acetate (0.2 mmol, 0.1 equiv.) were added. The reaction was heated to 90°C for 2 hours. After TLC confirmed complete conversion, the mixture was cooled to room temperature. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, washed with water and saturated brine, respectively, and dried over anhydrous sodium sulfate. Silica gel was added and the sample was separated by column chromatography to obtain the desired product 13 as a light yellow oily liquid. The total yield of the two-step reaction was 65%.

[0192] Step B: Preparation of compound 12ab

[0193] 96 mg of lithium hydroxide (4 mmol, 8 equiv.) and hydroxylamine aqueous solution (5 mmol, 10 equiv.) were weighed into a single-necked flask and dissolved in 2 mL of methanol. At -10°C, 182 mg of compound 13 (0.5 mmol, 1 equiv.) was dissolved in 3 mL of methanol and slowly added dropwise to the reaction solution. After stirring at -10°C for 30 minutes, the mixture was slowly warmed to room temperature and stirred for 8 hours. The reaction solution was then dried, 10 mL of water was added, and the pH was adjusted to 3-4 with 2 M hydrochloric acid. The mixture was filtered, and the residue was washed twice with water and once with ethyl acetate. The filter cake was collected and dried to obtain compound 12ab as a light yellow solid with a yield of 66%; mp 197.9-199.7°C; R f =0.3 (DCM:MeOH 20:1). 1H NMR (400MHz, DMSO-d6) δ11.14(s,1H),10.09(s,1H),8.98(s,1H),7.74(s,4H),7.63-7.44(m,2H),7.37-7.17(m,2H),3.88(s,2H), 3.49(s,2H),3.03-2.93(m,2H),2.81-2.69(m,2H). 13 C NMR(100MHz,DMSO-d6)δ169.4,164.4,154.5,151.5,141.7,128.1,127.9,124.1,123.2,119.3,111.5,60.6,50.1,49.8,20.6.HRMS(ESI)m / z Calcd for C 20 H 20 N3O4(M+H) + :366.1448;Found:366.1448.

[0194] Example 40: Preparation of Compound 12ac

[0195] Structural formula of compound 12ac

[0196] Step A: Synthesis of compound 4a

[0197] In a single-necked flask, 1.62 g of compound 1a (10 mmol, 1 equiv.), 3.03 g of paraformaldehyde (100 mmol, 10 equiv.), and 2.51 g of glycine methyl ester hydrochloride (20 mmol, 2 equiv.) were weighed and thoroughly dissolved in 20 mL of acetic acid. After stirring, the mixture was heated to 60°C and reacted for 3 hours. After TLC confirmed complete conversion, the mixture was cooled to room temperature and neutralized with saturated sodium bicarbonate solution. Ethyl acetate (3 × 100 mL) was then added for extraction. The organic phases were combined, washed once with saturated brine (1 × 100 mL), and dried over anhydrous sodium sulfate. Silica gel was added and the sample was mixed. Column chromatography was performed to obtain the desired product 4a as a white solid in a 76% yield.

[0198] Step B: Synthesis of compound 10a

[0199] In a single-necked flask, 550 mg of 4a (2 mmol, 1 equiv.) was weighed and dissolved in 4 mL of MeOH and 2 mL of water. 96 mg of lithium hydroxide (4 mmol, 2 equiv.) was added and allowed to react at room temperature for 8 h. The reaction solution was spin-dried, 10 mL of water was added, and the pH was adjusted to 1-2 with 4 M hydrochloric acid. The reaction solution was filtered, and the filter cake was washed with water and petroleum ether to obtain compound 10a as a white solid in 80% yield.

[0200] Step C: Preparation of compound 11ac

[0201] In a single-necked flask, 261 mg of compound 10a (1 mmol, 1 equiv.), 570 mg of HATU (1.5 mmol, 1.5 equiv.), and 178 mg of (E)-methyl 3-(4-aminophenyl)-2-propenoate (1 mmol, 1 equiv.) were weighed and thoroughly dissolved in 6 mL of DMF. 0.35 mL of DIPEA (2 mmol, 2 equiv.) was added and stirred at room temperature for 2 hours. The mixture was then heated to 60°C and allowed to react for 6 hours. After TLC confirmed complete conversion, 20 mL of water was added, and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, washed with saturated brine (5 × 20 mL), and dried over anhydrous sodium sulfate. Silica gel was added and the sample was then separated by column chromatography to yield the desired product 11ac as a pale yellow oil in a 67% yield.

[0202] Step D: Preparation of compound 12ac

[0203] In a single-necked flask, 96 mg of lithium hydroxide (4 mmol, 8 equiv.) and a hydroxylamine aqueous solution (5 mmol, 10 equiv.) were weighed and dissolved in 2 mL of methanol. At -10°C, 210 mg of compound 11ac (0.5 mmol, 1 equiv.) was dissolved in 3 mL of methanol and slowly added dropwise to the reaction mixture. After stirring at -10°C for 30 minutes, the mixture was slowly warmed to room temperature and stirred for 8 hours. The reaction mixture was then dried, 10 mL of water was added, and the pH was adjusted to 3-4 with 2M hydrochloric acid. The mixture was filtered, and the residue was washed twice with water and once with ethyl acetate. The filter cake was collected and dried to obtain compound 12ac as a white solid with a yield of 84%; mp 199.6-201.3°C; R f =0.3 (DCM:MeOH 10:1). 1H NMR (600MHz, DMSO-d6) δ10.70(s,1H),9.99(s,1H),8.99(s,1H),7.71(d,J=8.0Hz,2H),7.51(d,J=8.2Hz,2H),7.43-7.31(m,2H),7.15(d,J=2.2Hz,1 H),6.86(dd,J1=8.4Hz,J2=2.2Hz,1H),6.37(d,J=15.6Hz,1H),3.81(s,2H ),3.78(s,3H),3.44(s,2H),2.92(t,J=5.6Hz,2H),2.69(t,J=5.6Hz,2H). 13 C NMR(150MHz,DMSO-d6)δ169.2,163.5,157.5,155.4,150.3,140.3,138.4,130.3,128.6,12 1.4,120.1,119.3,118.0,111.4,111.2,96.9,60.7,56.0,50.1,49.8,20.6.HRMS(ESI)m / z Calcd for C 23 H 24 N3O5(M+H) + :422.1710; Found:422.1706.

[0204] According to the preparation method of Example 40, appropriate starting materials were selected to prepare the compound of Example 41.

[0205] Example 41: Preparation of Compound 12ad

[0206] The structural formula of compound 12ad

[0207] The preparation method is the same as that of Example 40, white solid, yield 62%; mp 151.8–155.1°C; R f =0.3 (DCM:MeOH 10:1). 1H NMR (400MHz, DMSO-d6) δ10.73(s,1H),10.14(s,1H),9.03(s,1H),7.62(d,J=8.0Hz,2H),7.50(d,J=8.2Hz,2H),7.44-7.33(m,2H),7.33-7.23(m ,4H),7.23-7.11(m,2H),6.89-6.81(m,1H),6.37(d,J=15.8Hz,1H),4.0 5-3.73(m,6H),3.29-3.17(m,1H),3.10-2.94(m,3H),2.72-2.55(m,2H). 13 C NMR (100MHz, DMSO) δ169.8,163.5,157.5,155.5,140.2,139.2,138.4,130.4,129.7,128.7,128.6, 126.7,121.4,120.0,119.3,118.0,111.4,96.9,69.5,56.1,47.3,46.6,34.4,21.6.HRMS(ESI)m / z Calcd for C 30 H 30 N3O5(M+H) + :512.2180; Found:512.2174.

[0208] Example 42: In vitro anti-histone deacetylase 1 activity test of target compounds

[0209] The HDAC1 in vitro inhibitory enzyme activity screening platform was used, and the inhibition was detected using the Fluorescence assay method. The specific steps are as follows:

[0210] Prepare 1x assay buffer (modified Tris Buffer);

[0211] Dilute the test compound and transfer it to the test plate with 100% DMSO, repeat three times;

[0212] Prepare enzyme solution with 1x assay buffer;

[0213] Prepare substrate solution with trypsin and acetylated peptide substrate in assay buffer;

[0214] Transfer 15 μL of enzyme solution to the assay plate and incubate at room temperature for 15 minutes;

[0215] Add 10 μL of substrate solution to each well of the plate;

[0216] The absorbance was measured at wavelengths of 355 nm and 460 nm, and the HDAC1 inhibitory activity of the test compound was calculated.

[0217] Table 1 HDAC1 inhibitory activity of compounds

[0218] In the above table, “+++” represents 0-10 nM; “++” represents 10-100 nM; “+” represents 100-1000 nM; and “-” represents >1000 nM.

[0219] Example 43: In vitro anti-tumor cell proliferation activity test of target compounds

[0220] The MTT assay was used to test the in vitro antiproliferative activity of the target compounds against HepG2, Bel-7402, and Huh-7 cell lines. The cells were placed in RPMI-1640 medium containing 10% fetal bovine serum (FPS) and 5% penicillin-streptomycin and cultured in a 37°C incubator containing 5% CO2. Cells in the logarithmic growth cycle were plated at 5×10 cells per well. 3 The cells were seeded into 96-well plates at a density of 100 cells / well and placed in a 37°C incubator containing 5% CO2 for about 18-24 hours. According to the experimental design, 100 μL of compounds with different concentration gradients were added to each well. No compound was added to the blank control group. SAHA or Abexinostat with the same concentration gradient was added to the positive control group and cultured in the incubator for 72 hours. When the drug action time was reached, 20 μL of 5 mg / mL MTT solution was added to each well in the dark and incubated in the incubator for 4-6 hours. Subsequently, 100 μL of MTT triple buffer was added to each well and incubated in the dark for 12 hours. The 96-well plate was placed in a microplate reader and the absorbance was measured at 570 nm and 650 nm using the dual wavelength method to calculate the inhibition rate of the test compound.

[0221] Table 2 Hepatocellular carcinoma cell inhibitory activity of compounds

[0222] In the above table, “+++” represents 0.01-0.1 μM; “++” represents 0.1-1 μM; “+” represents 1-10 μM; “-” represents >10 μM; and “NT” represents not tested.

Claims

1. A 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine-containing hydroxamic acid compound, characterized in that: The compound represented by the general formula (1) and / or the formula (2) and its pharmaceutically acceptable salt and / or hydrate, deuterated product, and / or its racemic mixture, hydrate, solvate, prodrug, enantiomer, diastereomer and tautomer; Where R 1 , R 2 , R 3 , R 4 are the same or different, and are each independently selected from one or more of hydrogen, halogen, nitro, cyano, amino, hydroxyl, hydroxymethyl, hydroxyethyl, mercapto, carboxyl, ester, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, aromatic heteroyl, alkenyl, alkynyl, alkyl monosubstituted amino, alkyl disubstituted amino, alkoxy, alkylcarbonyloxy, alkylcarbonylamino, cycloalkylcarbonylamino, heteroarylcarbonylamino, aminocarbonyl, alkoxyformamido, alkylthio, hydroxyalkoxy, sugar residue, sulfonic acid, phosphoric acid, polyhydroxyalkoxycarbonyl, carboxyalkoxy, carboxyalkylformyloxy, or R 2 R 3 Connect with R 2 and R 3 The two C's of R form a substituted or unsubstituted aryl ring structure, or a substituted or unsubstituted heteroaryl ring structure, or a substituted or unsubstituted cycloalkyl structure, or a substituted or unsubstituted heteroatom-containing cycloalkyl; 5 For H. Where n is a natural number between 0 and 2. represents the junction site; L in formula (1) and / or formula (2) is selected from the following: Where n is a natural number from 0 to 8. Represents the attachment site of L.

2. The 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine-containing hydroxamic acid compound according to claim 1, characterized in that: The pharmaceutically acceptable salt is an inorganic acid salt or an organic acid salt; The inorganic salt is selected from the group consisting of salts formed by any one of the following inorganic acids: hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid; The organic acid salt is selected from the group consisting of salts formed by any one of the following organic acids: acetic acid, trifluoroacetic acid, malonic acid, citric acid, tartaric acid, pyruvic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, mandelic acid, lactic acid and p-toluenesulfonic acid.

3. The 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine-containing hydroxamic acid compound according to claim 1, characterized in that: Where R 1 , R 2 , R 3 , R 4 The same or different, each independently selected from hydrogen, halogen, nitro, cyano, amino, hydroxyl, hydroxymethyl, hydroxyethyl, mercapto, carboxyl, ester, substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heterocycloalkyl, C5-C 10 Aromatic hetero groups, C2-C8 alkenyl groups, C2-C8 alkynyl groups, C1-C8 alkyl monosubstituted amino groups, C1-C8 alkyl disubstituted amino groups, C1-C8 alkoxy groups, C1-C8 alkyl carbonyloxy groups, C1-C8 alkyl carbonylamino groups, C3-C8 cycloalkyl carbonylamino groups, C5-C 10 one or more of heteroarylcarbonylamino, aminocarbonyl, C1-C8 alkoxycarboxamide, C1-C8 alkylthiol, hydroxy C1-C8 alkoxy, sugar residue, sulfonic acid, phosphoric acid, polyhydroxy C1-C8 alkoxycarbonyl, carboxyl C1-C8 alkoxy, carboxyl C1-C8 alkylformyloxy, or R 2 R 3 Connect with R 2 and R 3 The two C's form a substituted or unsubstituted C6~C 10 The ring structure of the aromatic group, or the formation of substituted or unsubstituted C5~C 10 The ring structure of the heteroaryl group may form a substituted or unsubstituted C3-C8 cycloalkyl structure, or may form a substituted or unsubstituted C3-C8 cycloalkyl group containing a heteroatom.

4. The 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine-containing hydroxamic acid compound according to claim 1, characterized in that: R 1 , R 2 , R 3 , R 4 are the same or different and are independently selected from hydrogen, C1-C8 alkyl, C1-C8 alkoxy, halogen, hydroxyl, or R 2 , R 3 Connect with R 2 and R 3 The two C's together form a heterocycloalkyl group, wherein the heterocycloalkyl group is a 3-8 membered ring having 1 or 2 selected from N, O, and S; R 5 For H. Where n is a natural number between 0 and 2. represents the junction site; L in formula (1) and / or formula (2) is selected from the following: Where n is a natural number from 0 to 8. Represents the attachment site of L.

5. The 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine-containing hydroxamic acid compound according to claim 1, characterized in that: The 1,2,3,4-tetrahydrobenzo[4,5]furan[2,3-C]pyridine hydroxamic acid compound is selected from any one of the following compounds or pharmaceutically acceptable salts and / or hydrates, deuterated compounds, and / or racemic mixtures, hydrates, solvates, prodrugs, enantiomers, diastereomers and tautomers thereof:

6. The method for preparing the compound according to claim 5, characterized in that: Compounds 5a-5g were synthesized by reaction route 1, compounds 8a-8c were synthesized by reaction route 2, compound 8d was synthesized by reaction route 3, compounds 12a-12aa were synthesized by reaction route 4, compound 12ab was synthesized by reaction route 5, and compounds 12ac and 12ad were synthesized by reaction route 6; Reaction Scheme 1 The process includes: Compounds 1, 2 and 3a-3g undergo Mannich reaction to generate compounds 4a-4g, which are then reacted with hydroxylamine to obtain compounds 5a-5g; Reaction Scheme 2 The process includes: Compounds 1a, 2 and 3h undergo Mannich reaction to generate compound 4h, which is then debenzylated with hydrogen under the catalysis of palladium carbon to obtain compound 6, which is then condensed with the corresponding carboxylic acid to obtain compounds 7a-7c, and finally reacted with hydroxylamine to obtain compounds 8a-8c; Reaction Scheme 3 The process includes: Compounds 1a, 2 and 3i undergo Mannich reaction to generate compound 4i, which is then substituted with methyl p-hydroxybenzoate to obtain an ester intermediate, which reacts with hydroxylamine to obtain compound 8d; Reaction Scheme 4 The process includes: Compounds 1, 2 and 3 undergo a Mannich reaction to obtain compound 4 or compound 9, which is then hydrolyzed to the corresponding carboxylic acid compound 10 under the action of lithium hydroxide, followed by condensation with methyl p-aminobenzoate to form compounds 11a-11aa, which are then reacted with hydroxylamine to obtain compounds 12a-12aa; Reaction Scheme 5 The process includes: Compound 11ab reacts with trifluoromethanesulfonic anhydride to obtain an intermediate, which is then reacted with formic acid under the catalysis of palladium acetate to generate compound 13, which is then reacted with hydroxylamine to obtain compound 12ab; Reaction Scheme 6 includes the following process: Compound 10 is condensed with the corresponding amine to give compounds 11ac and 11ad, which are then reacted with hydroxylamine to give compounds 12ac and 12ad.

7. A pharmaceutical composition, characterized in that Comprising the compound according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier thereof.

8. Use of a compound according to any one of claims 1 to 5 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or a pharmaceutical composition according to claim 7 in the preparation of a drug for treating a disease associated with histone deacetylase.

9. The use according to claim 8, characterized in that: The effects of the drug include, but are not limited to, anti-tumor, anti-inflammatory and neuroprotective.

10. Use of a compound according to any one of claims 1 to 5 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, or a pharmaceutical composition according to claim 7 in the preparation of a drug for treating a related disease, characterized in that: The medicine is a medicine for preventing and / or treating liver cancer, leukemia, multiple myeloma, lymphoma, head and neck cancer, esophageal cancer, prostate cancer, colon cancer, pancreatic cancer, lung cancer, stomach cancer, breast cancer and ovarian cancer.

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