Preparation of compound with effect of increasing content of lanosterol in brain, pharmaceutical composition of compound and application of compound

Abstract

The invention belongs to the technical field of medicines, and discloses a preparation method of a series of compounds which have the effect of increasing the content of lanosterol in the brain and have general formulas (I) and (II) as well as a pharmaceutical composition and application of the compounds.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of medicine, and relates to a compound with the effect of increasing the content of lanosterol in the brain, a preparation method thereof, a pharmaceutical composition containing the same, and the use of the compound as a medicine, especially as an anti-neurodegenerative disease such as Parkinson's Disease (PD) and Alzheimer's Disease (AD), for improving learning and memory impairment, and for treating memory function decline. BACKGROUND

[0002] Parkinson's Disease (PD) is the fastest growing neurological disease, and its incidence ranks second among neurodegenerative diseases (Lancet Neurol 2018; 17: 939-953). The main pathological features of PD are the progressive degeneration of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies (Redox Bio 2017; 11: 606-612). Epidemiological studies have shown that the prevalence of PD in people over 60 years old in Europe and the United States is as high as 1%, and in people over 80 years old, it is more than 4%. In China, the prevalence rate of people over 65 years old is 1.7%, which is similar to that in Europe and the United States. With the aggravation of population aging, it is estimated that the number of PD patients in the world will reach 120 million by 2050 (Lancet Neurol 2018; 17: 928-929). With the progression of the disease, the motor and non-motor symptoms of PD gradually worsen, not only affecting the quality of life of patients, but also bringing huge social and economic burden.

[0003] Currently, the drugs for PD treatment mainly improve the level of dopamine in the brain, which can be divided into three categories according to the function. The first category is the dopamine supplement drug, such as levodopa (combined with carbidopa), which is a dopamine precursor drug that can pass through the blood-brain barrier and generate dopamine after decarboxylation by decarboxylase. A study published in the New England Journal of Medicine recently showed that levodopa cannot alleviate the progression of the disease and is a drug that treats the symptoms rather than the root cause (N Engl J Med 2019; 380: 315-324). In addition, long-term use of levodopa can also produce a series of side effects, such as dopamine-induced movement disorders (JAMA Neurol 2017; 74: 941-949). The second category is the dopamine metabolism inhibitor, such as selegiline, which is an irreversible inhibitor of monoamine oxidase type B (MAO-B) (J Neural Transm Suppl, 1986; 22: 91-105). MAO-B can cause dopamine metabolism in the brain, i.e., deamination caused by oxidation. Selegiline binds to MAO-B in the substantia nigra, thereby inhibiting the binding and metabolism of dopamine by the enzyme. Benztropine hydrochloride can bind to acetylcholine receptors, indirectly causing an increase in dopamine release. The third category is the dopamine receptor agonist, such as pramipexole, ropinirole, and apomorphine, which can compensate for the decrease in dopamine levels by agonizing dopamine receptors. Other commonly used drugs also include amantadine, which can improve the symptoms of the disease, but long-term use can cause edema and confusion. It can be seen that the current drugs for PD treatment have a single target and cannot change the progression of the disease. Therefore, it is a major requirement in clinical practice to discover drug targets and drugs that can change the progression of PD.

[0004] Lanosterol (32) is a sterol compound, which is also the first sterol intermediate in the synthesis of cholesterol, and its main target is CYP51A1. At present, there are many studies on lanosterol, but most of them are about the relationship between lanosterol and antifungal activity. However, in 2011, researchers found that lanosterol has a dopaminergic neuron protective effect (Cell Death and Differentiation 2012; 19: 416-427). Since then, the related effects of intracerebral lanosterol have been gradually explored, which not only can protect neurons in the brain and inhibit neuroinflammation (Molecular Neurodegeneration 2015; 10: 48-65), but also has a clearing effect on amyloid (ACS Chem. Neurosci 2019; 10: 4051-4060). Although lanosterol has strong neuroprotective effect, direct administration has many inconveniences, such as short half-life, poor solubility, limited administration method (J Chem Eng Data 2020; 65: 436-445) (J Med Chem 2018; 61: 8693-8706) and so on.

[0005] Therefore, it is urgent to develop a compound that can increase the content of intracerebral lanosterol for the treatment of PD and other neurodegenerative diseases.

[0006] SUMMARY

[0007] A pharmaceutical composition, characterized in that it contains a compound according to any one of claims 1-12 or a salt thereof or an isomer thereof as an active ingredient and a pharmaceutically acceptable carrier.

[0008] The pharmaceutical composition according to claim 13, characterized in that it is selected from tablets, capsules, pills or injections.

[0009] Use of a compound according to any one of claims 1-12 or a salt thereof or an isomer thereof in the preparation of a medicament for the prevention and / or treatment of neurodegenerative diseases.

[0010] The use according to claim 15, characterized in that the neurodegenerative disease is selected from Parkinson's disease.

[0011] Use of a compound according to any one of claims 1-12 or a salt thereof or an isomer thereof in the preparation of a medicament for improving learning and memory disorders or for the treatment of memory impairment and Alzheimer's disease.

[0012] In summary, in order to overcome the shortcomings of the prior art, the present application aims to provide a compound with the effect of increasing the content of lanosterol in the brain;

[0013] Another object of the present application is to provide a preparation method of a compound with the effect of increasing the content of lanosterol in the brain.

[0014] Another object of the present application is to provide a compound with the effect of increasing the content of lanosterol in the brain and a composition thereof as a therapeutic drug for treating neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease (AD).

[0015] Still another object of the present application is to provide the use of a compound with the effect of increasing the content of lanosterol in the brain in a medicament for improving learning and memory disorders or in a therapeutic drug for memory decline.

[0016] Specifically, the present application relates to a compound of general formula (I) or a pharmaceutically acceptable salt thereof or a corresponding isomer thereof.

[0017]

[0018] characterized in that:

[0019] A ring is a benzene ring or an aromatic heterocycle containing 3-6 carbon atoms;

[0020] X is an alkylene group having 1-5 carbon atoms or an alkylene group containing a carbonyl group having 1-5 carbon atoms;

[0021] R1, R2 can be located at any position of the A ring, respectively;

[0022] R1, R2 are independently selected from hydrogen, a linear or branched alkyl group having 1-5 carbon atoms with or without oxygen, a linear or branched alkyl group having 1-5 carbon atoms substituted by halogen, nitro, cyano, acyloxy, sulfonyl, halogen.

[0023] and a compound of general formula (II) or a pharmaceutically acceptable salt thereof or a corresponding isomer thereof.

[0024]

[0025] characterized in that:

[0026] A, B, C ring is a benzene ring or an aromatic heterocycle containing 3-6 carbon atoms;

[0027] X can be O or N; n can be 0-5

[0028] R1, R2 can be located at any position of the A ring, respectively; R3, R4 can be located at any position of the B ring, respectively; R5, R6 can be located at any position of the C ring, respectively;

[0029] R1, R2, R3, R4, R5, R6are independently selected from hydrogen, linear or branched alkyl group with or without oxygen, halogen-substituted linear or branched alkyl group with 1-5 carbon atoms, nitro group, cyano group, acyloxy group, sulfonyl group, halogen; R7may be linear or branched alkyl group with or without oxygen, with 1-5 carbon atoms.

[0030] In addition, the present application also relates to a compound of general formula (Ia) or its pharmaceutically acceptable salt or its corresponding isomer.

[0031]

[0032] characterized in that:

[0033] A ring is a benzene ring;

[0034] X is alkylene with 1-5 carbon atoms and alkylene with 1-5 carbon atoms containing carbonyl group;

[0035] R1, R2may be located at any position of A ring respectively;

[0036] R1, R2are independently selected from hydrogen, linear or branched alkyl group with or without oxygen, halogen-substituted linear or branched alkyl group with 1-5 carbon atoms, nitro group, cyano group, acyloxy group, sulfonyl group, halogen.

[0037] and a compound of general formula (IIa) or its pharmaceutically acceptable salt or its corresponding isomer.

[0038]

[0039] A, B, C rings are benzene rings;

[0040] X can be O or N;

[0041] R1, R2may be located at any position of A ring respectively; R3, R4may be located at any position of B ring respectively; R5, R6may be located at any position of C ring respectively;

[0042] R1, R2, R3, R4, R5, R6are independently selected from hydrogen, linear or branched alkyl group with or without oxygen, halogen-substituted linear or branched alkyl group with 1-5 carbon atoms, nitro group, cyano group, acyloxy group, sulfonyl group, halogen.

[0043] According to the present application, preferred compounds of general formula (I), (II), (Ia), (IIa) include but are not limited to compounds represented by formula (Ia-1),

[0044]

[0045] wherein:

[0046] A ring is a benzene ring;

[0047] R1, R2may be located at any position of A ring respectively;

[0048] R1, R2are independently selected from hydrogen, linear or branched alkyl group with or without oxygen, halogen-substituted linear or branched alkyl group with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, halogen.

[0049] According to the present application, preferred compounds of general formula (I), (II), (Ia), (IIa) include but are not limited to compounds represented by formula (Ia-2),

[0050]

[0051] wherein:

[0052] A ring is a benzene ring;

[0053] R1, R2may be located at any position of A ring respectively;

[0054] R1, R2are independently selected from hydrogen, linear or branched alkyl group with or without oxygen, halogen-substituted linear or branched alkyl group with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, halogen.

[0055] According to the present application, preferred compounds of general formula (I), (II), (Ia), (IIa) include but are not limited to compounds represented by formula (Ia-3),

[0056]

[0057] wherein:

[0058] A ring and A" ring are benzene rings;

[0059] R1, R2may be located at any position of A ring respectively;

[0060] R1, R2are independently selected from hydrogen, linear or branched alkyl group with or without oxygen, halogen-substituted linear or branched alkyl group with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, halogen;

[0061] R1", R2"may be located at any position of A ring respectively;

[0062] R1", R2"are independently selected from hydrogen, linear or branched alkyl group with or without oxygen, halogen-substituted linear or branched alkyl group with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, halogen.

[0063] According to the present application, preferred compounds of general formula (I), (II), (Ia), (IIa) include but are not limited to compounds represented by formula (II-1),

[0064]

[0065] wherein:

[0066] A ring is a benzene ring or an aromatic heterocycle containing 3-6 carbon atoms; B, C ring is a benzene ring;

[0067] X can be O or N;

[0068] R1, R2 can be located at any position of A ring respectively; R3, R4 can be located at any position of B ring respectively; R5, R6 can be located at any position of C ring respectively;

[0069] R1, R2, R3, R4, R5, R6 are independently selected from hydrogen, linear or branched alkyl containing oxygen or not containing oxygen with carbon atoms number of 1-5, halogen-substituted linear or branched alkyl with carbon atoms number of 1-5, nitro, cyano, acyloxy, sulfonyl, halogen.

[0070] According to the present application, preferred compounds of general formula (I), (II), (Ia), (IIa) include but are not limited to compounds represented by formula (II-2),

[0071]

[0072] wherein:

[0073] B ring is a benzene ring or an aromatic heterocycle containing 3-6 carbon atoms; A, C ring is a benzene ring;

[0074] X can be O or N;

[0075] R1, R2 can be located at any position of A ring respectively; R3, R4 can be located at any position of B ring respectively; R5, R6 can be located at any position of C ring respectively;

[0076] R1, R2, R3, R4, R5, R6 are independently selected from hydrogen, linear or branched alkyl containing oxygen or not containing oxygen with carbon atoms number of 1-5, halogen-substituted linear or branched alkyl with carbon atoms number of 1-5, nitro, cyano, acyloxy, sulfonyl, halogen.

[0077] According to the present application, preferred compounds of general formula (I), (II), (Ia), (IIa) include but are not limited to compounds represented by formula (II-3),

[0078]

[0079] wherein:

[0080] Rings A and B are benzene rings;

[0081] X can be 0 or N; n can be 0-5;

[0082] R1 and R2 can be located at any position in ring A; R3 and R4 can be located at any position in ring B.

[0083] R1, R2, R3, and R4 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

[0084] According to the present invention, preferred compounds of general formulas (I), (II), (Ia), and (IIa) include, but are not limited to, compounds represented by formula (II-4).

[0085]

[0086] in:

[0087] Rings A and C are benzene rings;

[0088] X can be O or N;

[0089] R1 and R2 can be located at any position in ring A; R5 and R6 can be located at any position in ring C.

[0090] R1, R2, R5, and R6 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

[0091] The most preferred compound of this invention is:

[0092]

[0093]

[0094]

[0095]

[0096]

[0097] A method for preparing the compound or its isomers according to the present invention includes the following steps:

[0098] 1. 2,3-Disubstituted benzaldehyde and substituted phenylacetic acid are condensed in the presence of an anhydride to obtain 2,3-disubstituted phenylacrylic acid, which is then condensed with p-substituted benzyloxyphenethylamine. If the benzene ring contains a protecting substituent, such as an acetoxy group, the protecting group can be removed by hydrolysis. Next, the hydroxyl group on the parent structure is selectively protected using a tert-butyloxycarbonyl group, followed by reaction with substituted benzyl bromide or 2-bromosubstituted phenylacetone. Finally, the tert-butyloxycarbonyl group protection is removed to obtain the target product.

[0099] Specifically, according to the present invention, the compounds of general formula (I)(Ia) represented by formulas (Ia-1, Ia-2, Ia-3) can be prepared via reaction route I:

[0100] Scheme I :

[0101]

[0102] i: condensation; ii: acylation; iii: hydrolysis; iv: tert-butyloxycarbonyl protection; v: condensation; vi: hydrolysis

[0103] Among them, the C ring is a benzene ring or an aromatic heterocycle containing heteroatoms;

[0104] X can be alkylene, oxyalkylene, carbonylalkylene, acyloxyalkylene, or amidealkylene;

[0105] 2. Methyl butyryneate is condensed with its corresponding iodide and borate in the presence of palladium catalyst and potassium carbonate to obtain a mixed α-structure. This is followed by ester hydrolysis under strongly alkaline conditions to yield the corresponding carboxylic acid compound. In this step, the isomers are separated to obtain cis and trans structures. Finally, dehydration condensation with the corresponding amino or hydroxyl compound yields the target product. If a protecting group is present, an additional protecting group removal reaction is performed after the reaction pathway is completed.

[0106] Specifically, according to the present invention, compounds of general formula (II)(IIa) represented by formulas (II-1, II-2, II-3, II-4) can be prepared via reaction route II:

[0107] Scheme II :

[0108]

[0109] i: condensation; ii: hydrolysis; iii: isomer separation; iv: condensation

[0110] Among them, rings A, B, and C are benzene rings or aromatic heterocycles containing heteroatoms;

[0111] X can be O or N.

[0112] In this invention, the term "alkyl" refers to a straight-chain or branched alkyl group containing 1-8 carbon atoms, such as ethyl, propyl, and n-butyl.

[0113] In this invention, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0114] According to the present invention, compounds of general formulas (I), (II), (Ia), and (IIa) may exist in the form of isomers, and the term "compound of the present invention" generally includes isomers of the compound.

[0115] Compounds of general formulas (I), (II), (Ia), and (IIa) may exist as cis-trans isomers with double bonds, and the asymmetric center has an S or R configuration. This invention includes all possible stereoisomers and mixtures of two or more isomers. If cis / trans isomers are present, this invention relates to cis and trans isomers and mixtures of these forms. If a single isomer is required, it can be isolated according to conventional methods or prepared by stereoselective synthesis.

[0116] According to the present invention, compounds of general formulas (I), (II), (Ia), and (IIa) and their isomers, when used in animals, preferably mammals, and especially humans, have shown excellent efficacy in treating neurodegenerative diseases such as Parkinson's disease (PD) and Alzheimer's disease (AD), improving learning and memory impairment, and treating memory decline. Therefore, they can be used as medicines for treating Parkinson's disease or preventing and treating learning and memory impairment, treating memory decline, and treating Alzheimer's disease.

[0117] The present invention therefore also relates to pharmaceutical compositions containing at least one compound of general formula (I), (II), (Ia), (IIa) and / or its stereoisomers as an active ingredient, and conventional pharmaceutical excipients or adjuvants. Typically, the pharmaceutical compositions of the present invention contain 0.1-90% by weight of a compound of general formula (I), (II), (Ia), (IIa) and / or its physiologically acceptable salts.

[0118] Pharmaceutical compositions may be prepared according to methods known in the art. For this purpose, if desired, compounds of general formulas (I), (II), (Ia), (IIa) and / or stereoisomers may be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants to prepare a suitable administration or dosage form for use as a human or veterinary medicine.

[0119] The compounds of general formulas (I), (II), (Ia), and (IIa) of this invention, or pharmaceutical compositions containing them, can be administered in unit doses via enteral or non-enteric routes, such as oral, intramuscular, subcutaneous, nasal, oral mucosa, skin, peritoneum, or rectum. Dosage forms include tablets, capsules, drops, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, lyophilized powder injections, etc., and can be conventional preparations, sustained-release preparations, controlled-release preparations, and various particulate delivery systems. Various carriers known in the art can be widely used to formulate unit doses into tablets. Examples of carriers include diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, and aluminum silicate; humectants and binders such as water, glycerin, polyethylene glycol, ethanol, propanol, starch paste, dextrin, syrup, honey, glucose solution, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, shellac, methylcellulose, potassium phosphate, and polyvinylpyrrolidone; disintegrants such as dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, methylcellulose, and ethylcellulose; disintegration inhibitors such as sucrose, tristearate, cocoa butter, and hydrogenated oil; absorption promoters such as quaternary ammonium salts and sodium dodecyl sulfate; and lubricants such as talc, silica, corn starch, stearates, boric acid, liquid paraffin, and polyethylene glycol. Tablets can be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets. To formulate the drug delivery unit into a pill, a wide variety of carriers known in the art can be used. Examples of carriers include diluents and absorbents such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, gelucire, kaolin, talc, etc.; binders such as gum arabic, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste, or flour paste, etc.; and disintegrants such as agar powder, dried starch, alginate, sodium dodecyl sulfate, methylcellulose, ethylcellulose, etc. To formulate the drug delivery unit into a suppository, a wide variety of carriers known in the art can be used. Examples of carriers include polyethylene glycol, lecithin, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, semi-synthetic glycerides, etc. To encapsulate the drug delivery unit, the active ingredient of general formula (I), (II), (Ia), (IIa) or its stereoisomers are mixed with the various carriers described above, and the resulting mixture is placed in a hard gelatin capsule or a soft capsule. Alternatively, the active ingredient of general formula (I), (II), (Ia), (IIa) or its stereoisomers can be formulated as microcapsules, suspended in an aqueous medium to form a suspension, or filled into hard capsules or formulated as an injectable preparation for use.To formulate the drug delivery unit into injectable formulations, such as solutions, emulsions, lyophilized powders for injection, and suspensions, all diluents commonly used in the art can be used, such as water, ethanol, polyethylene glycol, 1,3-propanediol, ethoxylated isostearyl alcohol, polyoxyethylene isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, etc. Additionally, to prepare isotonic injections, appropriate amounts of sodium chloride, glucose, or glycerol can be added to the injectable formulation. Furthermore, conventional solubilizers, buffers, pH adjusters, etc., can also be added.

[0120] In addition, colorants, preservatives, flavorings, tasters, sweeteners or other materials may be added to pharmaceutical preparations if necessary. Attached Figure Description

[0121] Figure 1 Protective effect of the compound on SH-SY5Y cells in an H2O2 injury model; * P < 0.05 vs. Feline group.

[0122] Figure 2 Effects of some compounds on lanosterol content in SH-SY5Y cells. ** P < 0.01, *** P < 0.001, compared with the DMSO group; & P < 0.05, compared with the fenoproline group.

[0123] Figure 3 Effect of compound 12 on lanosterol content in the brain of MPTP PD mice. **P < 0.01 vs. model mice; && P < 0.01 vs. mice in the fenprolene group.

[0124] Figure 4 Effect of test drug compound 12 on residence time on the rotator in MPTP PD mice. ### P < 0.001 vs. control group mice; *P < 0.05, ***P < 0.001 vs. model group mice; & P < 0.05 vs. mice in the fenprolene group.

[0125] Figure 5 Effect of test drug compound 12 on pole climbing score in MPTP PD mice. ### P < 0.001 vs. control group mice; *** P < 0.001 vs. model group mice.

[0126] Figure 6 Effect of test drug compound 12 on residence time on transptiles in A53T transgenic mice. ### P < 0.001 vs. wild-type mice; ***P < 0.001 vs. A53T transgenic mice.

[0127] Figure 7 Effects of compound 12 administration on pole-climbing behavior in A53T transgenic mice. ### P < 0.001 vs. wild-type mice, *P < 0.05, **P < 0.01 vs. A53T transgenic mice.

[0128] Figure 8 Effects of compound 12 administration on balance beam behavior in A53T transgenic mice. ### P < 0.001 vs. wild-type mice, *P < 0.01, ***P < 0.001 vs. A53T transgenic mice. & P < 0.05 vs. mice in the fenprolene group.

[0129] Figure 9 Effects of compound 12 administration on scopolamine-treated mice jumping platform. ### P < 0.001 vs. control group mice, *P < 0.05, **P < 0.01 vs. model group mice. Detailed Implementation

[0130] The following examples and drug activity experiments are used to further illustrate the present invention, but they do not imply any limitation on the present invention.

[0131] In this invention, the raw materials used are known compounds or compounds prepared by methods known to those skilled in the art. The preparation does not generate the compounds of this invention, but is a useful intermediate for the synthesis of compounds of general formulas (I), (II), (Ia), and (IIa).

[0132] Example of compound synthesis:

[0133] Step A:

[0134] Example 1:

[0135] 2000 mg (4.45 mmol) of (E)-2-(2,5-dimethoxyphenyl)-3-(4-hydroxy-3-methoxyphenyl)-N-(4-hydroxyphenylethyl)acrylamide and 272 mg (2.23 mmol) of DMAP were dissolved in 120 mL of DMF and heated to 60 °C with stirring. 971 mg of BOC anhydride (4.45 mmol) was dissolved in 10 mL of DMF and added in 5 portions, 45 minutes apart, and reacted for 3 hours after each addition. Stirring was stopped, and the mixture was extracted with 300 mL of ethyl acetate and 320 mL of saturated brine. The aqueous layer was extracted twice with 300 mL of ethyl acetate. The organic layers were combined, washed three times with 300 mL of brine, and concentrated under reduced pressure. Column chromatography (PE:EA = 4:1 to 5:3) was used for purification, yielding 1964 mg of compound (31), with a yield of 80.3%.

[0136]

[0137] 1 H NMR(DMSO-d6)δ9.21(s,1H),7.54(s,1H),7.16–7.09(m,1H),7.09–6.95(m,5H),6.78(d,1H),6.73–6.66(m,3 H),6.62(s,1H),3.72(s,3H),3.67(s,3H),3.40(s,3H),3.33(q,2H),2.55(s,1H),1.54(s,1H),1.49(s,9H).

[0138] HRMS(ESI+)calcd for C 31 H 36 NO8 + (M+H + )550.2441,found 550.2425.

[0139] Step B:

[0140] Example 2:

[0141] 150 mg (0.27 mmol) of compound 31 was dissolved in 7.5 mL of acetonitrile, and 60 mg of anhydrous K₂CO₃ and 59 mg of 4-nitrobenzyl bromide were added. The mixture was heated to 60 °C and stirred. After reacting for 8 hours, stirring was stopped, K₂CO₃ was filtered off, the acetonitrile was evaporated to dryness, dissolved in 1 mL of dichloromethane, and purified by column chromatography (PE:EA = 1:1). 182.3 mg of the product was given, with a yield of 97.5%.

[0142] Dissolve 150 mg (0.22 mmol) of the product from the previous step in 3 mL of dichloromethane, add 900 μL of trifluoroacetic acid, stir at room temperature for 2 hours, and evaporate to dryness. 82 mg of product (3) was obtained, with a yield of 64.0%.

[0143]

[0144] 1H NMR(Chloroform-d)δ8.27(d,2H),7.80(s,1H),7.63(d,2H),7.03(d,2H),6.97–6.92(m,2H),6.85(d,2H),6.80–6.72( m,2H),6.67(s,1H),6.47(s,1H),5.64(s,1H),5.17(s,2H),3.73(s,3H),3.66(s,3H),3.59–3.48(m,5H),2.76(t,2H).

[0145] HRMS(ESI+)calcd for C 33 H 33 N2O8 + (M+H + )585.2237,found 585.2265.

[0146] Example 3:

[0147] Using a method similar to that in Example 2, compound 31 was reacted with 3-nitrobenzyl bromide to give compound 1 below:

[0148]

[0149] Yield: 62.8%

[0150] 1 H NMR(Chloroform-d)δ8.37(tt,1H),8.24(ddd,1H),7.85–7.77(m,2H),7.62(t,1H),7.08–7.01(m,2H),6.95(d,2H),6.91–6.84(m,2H),6.8 0–6.75(m,2H),6.68(t,1H),6.48(d,1H),5.73(s,1H),5.61(t,1H),5 .17(s,2H),3.74(s,3H),3.66(s,3H),3.60–3.50(m,5H),2.76(t,2H).

[0151] HRMS(ESI+)calcd for C 33 H 33 N2O8 + (M+H + )585.2237, found 585.2210.

[0152] Example 4:

[0153] Using a method similar to that in Example 2, compound 31 was reacted with 2-nitrobenzyl bromide to give compound 2 as shown below:

[0154]

[0155] Yield: 48.9%

[0156] 1 H NMR(Chloroform-d)δ8.21(dd,1H),7.92(dq,1H),7.81(s,1H),7.73(td,1H),7.54(tdd,1H),7.09–6.91(m,4H),6.91–6.84(m,2H),6. 80–6.75(m,2H),6.72–6.64(m,1H),6.48(s,1H),5.60(t,1H),5.50(s,2H),3.74(s,3H),3.66(s,3H),3.60–3.52(m,5H),2.76(t,2H).

[0157] HRMS(ESI+)calcd for C 33 H 33 N2O8 + (M+H + )585.2237, found 585.2236.

[0158] Example 5:

[0159] Using a method similar to that in Example 2, compound 31 was reacted with 3-fluorobenzyl bromide to give compound 4 below:

[0160]

[0161] Yield: 66.0%

[0162] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.40(td,1H),7.27–7.16(m,2H),7.13–6.98(m,3H),6.98–6.81(m,4H),6.81–6.73( m,2H),6.68(d,1H),6.49(s,1H),5.61(t,1H),5.07(s,2H),3.74(s,3H),3.64(s,3H),3.60–3.50(m,5H),2.75(t,2H).

[0163] HRMS(ESI+)calcd for C 33 H 33 FNO6 + (M+H +)558.2292,found 558.2290.

[0164] Example 6:

[0165] Using a method similar to that in Example 2, compound 31 was reacted with 3-chlorobenzyl bromide to give compound 5 below:

[0166]

[0167] Yield: 73.3%

[0168] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.48(s,1H),7.38–7.33(m,3H),7.06–6.81(m,6H),6.80–6.75(m,2H),6. 68(d,1H),6.49(s,1H),5.62(t,1H),5.05(s,2H),3.74(s,3H),3.64(s,3H),3.60–3.51(m,5H),2.75(t,2H).

[0169] HRMS(ESI+)calcd for C 33 H 33 ClNO6 + (M+H + )574.1996, found 574.1995.

[0170] Example 7:

[0171] Using a method similar to that in Example 2, compound 31 was reacted with 3-bromobenzyl bromide to give compound 6 below:

[0172]

[0173] Yield: 67.0%

[0174] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.64(t,1H),7.51(ddd,1H),7.39(d,1H),7.33–7.31(m,1H),7.06–6.81(m,6H),6.81–6 .70(m,2H),6.68(d,1H),6.51–6.46(m,1H),5.61(t,1H),5.04(s,2H),3.74(s,3H),3.63(s,3H),3.54(s,5H),2.75(t,2H).

[0175] HRMS(ESI+)calcd for C 33 H 33 BrNO6 + (M+H + )618.1491,found 618.1479.

[0176] Example 8:

[0177] Using a method similar to that in Example 2, compound 31 was reacted with 3-iodobenzyl bromide to give compound 7 below:

[0178]

[0179] Yield: 59.6%

[0180] 1 H NMR(Chloroform-d)δ7.85(t,1H),7.81(s,1H),7.71(ddd,1H),7.43(ddd,1H),7.17(t,1H),7.06–6.98(m,2H),6.98–6.89(m,2H),6.89–6.81( m,2H),6.81–6.73(m,2H),6.68(d,1H),6.51–6.46(m,1H),5.61(t,1H), 5.01(s,2H),3.74(s,3H),3.63(s,3H),3.60–3.50(m,5H),2.75(t,2H).

[0181] HRMS(ESI+)calcd for C 33 H 33 INO6 + (M+H + )666.1353,found 666.1336.

[0182] Example 9:

[0183] Using a method similar to that in Example 2, compound 31 was reacted with 3-methoxybenzyl bromide to give compound 8 below:

[0184]

[0185] Yield: 56.2%

[0186] 1H NMR(Chloroform-d)δ7.81(s,1H),7.40(s,4H),6.92(m,6H),6.78(m,2H),6.68(d,1H),6. 48(s,1H),5.61(t,1H),5.04(s,2H),3.73(s,3H),3.64(s,3H),3.54(m,5H),2.75(t,2H).

[0187] HRMS(ESI+)calcd for C 33 H 33 ClNO6 + (M+H + )574.1996, found 574.1977.

[0188] Example 10:

[0189] Using a method similar to that in Example 2, compound 31 was reacted with 2-chlorobenzyl bromide to give compound 9 below:

[0190]

[0191] Yield: 69.8%

[0192] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.59(m,1H),7.46(m,1H),7.33(m,2H),7.01(m,2H),6.91(m,4H),6.78(m, 2H),6.68(d,1H),6.49(s,1H),5.61(t,1H),5.18(s,2H),3.74(s,3H),3.64(s,3H),3.55(m,5H),2.76(t,2H).

[0193] HRMS(ESI+)calcd for C 33 H 33 ClNO6 + (M+H + )574.1996, found 574.1982.

[0194] Example 11:

[0195] Using a method similar to that in Example 2, compound 31 was reacted with 2,4-dichlorobenzyl bromide to give compound 10 as follows:

[0196]

[0197] Yield: 87.4%

[0198] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.52(d,1H),7.48(d,1H),7.32(dd,1H),7.02(m,2H),6.95(m,2H),6.86(m,2H),6 .78(m,2H),6.68(d,1H),6.49(s,1H),5.60(t,1H),5.13(s,2H),3.74(s,3H),3.65(s,3H),3.54(m,5H),2.76(t,2H).

[0199] HRMS(ESI+)calcd for C 33 H 33 Cl2NO6 + (M+H + )608.1607,found 608.1608.

[0200] Example 12:

[0201] Using a method similar to that in Example 2, compound 31 was reacted with 3,4-dichlorobenzyl bromide to give compound 11 below:

[0202]

[0203] Yield: 65.4%

[0204] 1 H NMR(Chloroform-d)δ7.77(s,1H),7.54(d,1H),7.46(d,1H),7.24(dd,1H),6.93(m,4H),6.76(m,4H),6 .63(d,1H),6.44(s,1H),5.57(t,1H),4.98(s,2H),3.69(s,3H),3.60(s,3H),3.50(m,5H),2.71(t,2H).

[0205] HRMS(ESI+)calcd for C 33 H 33 Cl2NO6 + (M+H + )608.1607,found 608.1598.

[0206] Example 13:

[0207] Using a method similar to that in Example 2, compound 31 was reacted with 3,5-dichlorobenzyl bromide to give compound 12 as follows:

[0208]

[0209] Yield: 83.3%

[0210] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.37(s,3H),7.02(m,2H),6.94(m,2H),6.81(m,4H),6.68(d,1H),6.49 (s,1H),5.60(t,1H),5.02(s,2H),3.74(s,3H),3.65(s,3H),3.54(m,5H),2.76(t,2H).HRMS(ESI+)calcd for C 33 H 33 Cl2NO6 + (M+H + )608.1607,found608.1593.

[0211] Example 14:

[0212] Using a method similar to that in Example 2, compound 31 was reacted with 2,5-dichlorobenzyl bromide to give compound 13 below:

[0213]

[0214] Yield: 82.2%

[0215] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.62(d,1H),7.38(d,1H),7.28(dd,1H),7.03(m,2H),6.91(m,4H),6.78(m, 2H),6.68(dd,1H),6.49(s,1H),5.61(t,1H),5.13(s,2H),3.74(s,3H),3.65(s,3H),3.56(m,5H),2.76(t,2H).

[0216] HRMS(ESI+)calcd for C 33 H 33 Cl2NO6 + (M+H + )608.1607, found 608.1605.

[0217] Example 15:

[0218] Using a method similar to that in Example 2, compound 31 was reacted with 3,5-difluorobenzyl bromide to give compound 14 below:

[0219]

[0220] Yield: 72.1%

[0221] 1 H NMR(DMSO-d6)δ9.27(s,1H),7.41(s,1H),7.18(m,3H),7.05(t,3H),6.96(dd,1H),6.90(d,2H),6.73(t,1H),6.62 (s,2H),6.56(d,1H),6.40(s,1H),5.11(s,2H),3.66(s,3H),3.59(s,3H),3.35(s,3H),3.29(m,2H),2.64(t,2H).

[0222] HRMS(ESI+)calcd for C 33 H 33 F2NO6 + (M+H + )576.2198, found 576.2182.

[0223] Example 16:

[0224] Using a method similar to that in Example 2, compound 31 was reacted with 4-tert-butylbenzyl bromide to give compound 15 below:

[0225]

[0226] Yield: 17.4%

[0227] 1 H NMR(Chloroform-d)δ7.77(s,1H),7.43(d,2H),7.37(d,2H),6.95(d,2H),6.86(m,4H),6.74(s,2H),6.63(d, 1H),6.44(s,1H),5.57(t,1H),4.99(s,2H),3.69(s,3H),3.57(s,3H),3.51(m,5H),2.70(t,2H),1.33(s,9H).

[0228] HRMS(ESI+)calcd for C 37 H 42 NO6 + (M+H + )596.3012,found 596.3007.

[0229] Example 17:

[0230] Using a method similar to that in Example 2, compound 31 was reacted with 4-n-butylbenzyl bromide to give compound 16 below:

[0231]

[0232] Yield: 44.1%

[0233] 1 H NMR(Chloroform-d)δ7.76(s,1H),7.34(d,2H),7.21(d,2H),6.95(d,2H),6.85(m,4H),6.73(s,2H),6.63(d,1H),6.45(s,1H),5. 55(t,1H),4.99(s,2H),3.69(s,3H),3.58(s,3H),3.50(m,5H),2.70(t,2H),2.62(t,2H),1.61(p,2H),1.37(h,2H),0.93(t,3H).

[0234] HRMS(ESI+)calcd for C 37 H 42 NO6 + (M+H + )596.3012,found 596.3009.

[0235] Example 18:

[0236] Using a method similar to that in Example 2, compound 31 was reacted with 3-methylbenzyl bromide to give compound 17 below:

[0237]

[0238] Yield: 53.3%

[0239] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.34(t,1H),7.28(t,2H),7.19(d,1H),6.99(m,2H),6.89(m,4H),6.78(m,2H),6. 68(d,1H),6.49(s,1H),5.62(t,1H),5.03(s,2H),3.74(s,3H),3.63(s,3H),3.55(m,5H),2.75(t,2H),2.43(s,3H).

[0240] HRMS(ESI+)calcd for C 34 H 36 NO6 +(M+H + )554.2543,found 554.2528.

[0241] Example 19:

[0242] Using a method similar to that in Example 2, compound 31 was reacted with 3-ethylbenzyl bromide to give compound 18 as shown below:

[0243]

[0244] Yield: 15.6%

[0245] 1 H NMR(Chloroform-d)δ7.76(s,1H),7.27(m,4H),7.18(d,1H),6.96(d,2H),6.86(m,4H),6.73(s,2H),6.63(d, 1H),6.45(s,1H),5.56(t,1H),5.00(s,2H),3.69(s,3H),3.59(s,3H),3.50(m,5H),2.69(m,4H),1.26(t,3H).

[0246] HRMS(ESI+)calcd for C 35 H 38 NO6 + (M+H + )568.2699,found 568.2701.

[0247] Example 20:

[0248] Using a method similar to that in Example 2, compound 31 was reacted with 3-trifluoromethylbenzyl bromide to give compound 19 below:

[0249]

[0250] Yield: 76.6%

[0251] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.75(s,1H),7.65(t,2H),7.56(t,1H),7.03(m,2H),6.90(m,4H),6.78(m, 2H),6.68(d,1H),6.49(s,1H),5.61(t,1H),5.12(s,2H),3.74(s,3H),3.65(s,3H),3.54(m,5H),2.76(t,2H).

[0252] HRMS(ESI+)calcd for C 34 H 33 F3NO6 + (M+H + )608.2260, found 608.2236.

[0253] Example 21:

[0254] Using a method similar to that in Example 2, compound 31 was reacted with 3-cyanobenzyl bromide to give compound 20 as follows:

[0255]

[0256] Yield: 77.5%

[0257] 1 H NMR(Chloroform-d)δ7.81(s,1H),7.79(td,1H),7.68(ddt,2H),7.55(t,1H),7.04(m,2H),6.95(d,2H),6.85(m,2H), 6.78(m,2H),6.68(t,1H),6.48(d,1H),5.61(t,1H),5.10(s,2H),3.74(s,3H),3.66(s,3H),3.54(m,5H),2.76(t,2H).

[0258] HRMS(ESI+)calcd for C 34 H 33 N2O6 + (M+H + )565.2339, found 565.2335.

[0259] Example 22:

[0260] Compound 31 was reacted with 3-acetylbenzyl bromide using a method similar to that in Example 2 to give compound 21 as follows:

[0261]

[0262] Yield: 79.8%

[0263] 1H NMR(Chloroform-d)δ8.03(t,1H),7.92(dt,1H),7.76(s,1H),7.65(d,1H),7.50(t,1H),6.98(m,2H),6.86(m,4H),6.73(m, 2H),6.63(dd,1H),6.44(s,1H),5.57(t,1H),5.08(s,2H),3.69(s,3H),3.60(m,3H),3.50(s,5H),2.71(t,2H),2.63(s,3H).

[0264] HRMS(ESI+)calcd for C 35 H 36 NO7 + (M+H + )582.2492,found 582.2478.

[0265] Example 23:

[0266] Using a method similar to that in Example 2, compound 31 was reacted with 3-methoxybenzyl bromide to give compound 22 as follows:

[0267]

[0268] Yield: 42.6%

[0269] 1 H NMR(Chloroform-d)δ7.80(s,1H),7.35(t,1H),7.03(m,4H),6.89(m,5H),6.78(m,2H),6.68(d,1H),6. 49(s,1H),5.61(t,1H),5.05(s,2H),3.87(s,3H),3.74(s,3H),3.62(s,3H),3.54(m,5H),2.74(t,2H).

[0270] HRMS(ESI+)calcd for C 34 H 36 NO7 + (M+H + )570.2492,found 570.2490.

[0271] Example 24:

[0272] Using a method similar to that in Example 2, compound 31 was reacted with 2-chloro-1-acetophenone to give compound 23 as shown below:

[0273]

[0274] Yield: 20.5%

[0275] 1 H NMR(Chloroform-d)δ8.01(m,2H),7.75(s,1H),7.63(ddt,1H),7.52(m,2H),6.92(m,4H),6.76(m,4H),6.63(t ,1H),6.44(s,1H),5.69(s,1H),5.55(t,1H),5.24(s,2H),3.68(s,3H),3.59(s,3H),3.49(m,5H),2.70(t,2H).

[0276] HRMS(ESI+)calcd for C 34 H 34 NO7 + (M+H + )568.2335,found 568.2338.

[0277] Example 25:

[0278] Using a method similar to that in Example 2, compound 31 was reacted with 2-chloro-1-(4-methoxyphenyl)ethyl ketone to give compound 24 below:

[0279] Yield: 54.5%

[0280] 1 H NMR(Chloroform-d)δ8.01(dt,2H),7.75(s,1H),6.96(m,4H),6.89(d,2H),6.76(m,4H),6.62(t,1H),6.44(s, 1H),5.68(s,1H),5.55(t,1H),5.18(s,2H),3.89(s,3H),3.69(s,3H),3.58(s,3H),3.49(m,5H),2.69(t,2H).

[0281] HRMS(ESI+)calcd for C 35 H 36 NO8 + (M+H + )598.2441,found 598.2442.

[0282] Example 26:

[0283] Using a method similar to that in Example 2, compound 31 was reacted with 2-chloro-1-(4-trifluoromethylphenyl)ethyl ketone to give compound 25 below:

[0284]

[0285] Yield: 47.9%

[0286] 1 H NMR(Chloroform-d)δ8.13(d,2H),7.77(d,3H),6.98(d,2H),6.90(d,2H),6.79(d,2H),6.73(s,2H),6. 63(d,1H),6.44(s,1H),5.56(t,1H),5.22(s,2H),3.69(s,3H),3.62(s,3H),3.50(m,5H),2.71(t,2H).

[0287] HRMS(ESI+)calcd for C 35 H 33 F3NO7 + (M+H + )636.2209, found 636.2211.

[0288] Example 27:

[0289] Using a method similar to that in Example 2, compound 31 was reacted with 2-chloro-1-(4-cyanophenyl)ethyl ketone to give compound 26 below:

[0290] Yield: 46.5%

[0291] 1 H NMR(Chloroform-d)δ8.10(d,2H),7.80(d,2H),7.75(s,1H),6.99(d,2H),6.80(m,6H),6.63(s,1H),6.43 (s,1H),5.56(t,1H),5.19(s,2H),3.69(s,3H),3.61(s,3H),3.50(m,5H),2.70(t,2H).HRMS(ESI+)calcd for C 35 H 33 N2O7 + (M+H + )593.2288,found 593.2296.

[0292] Example 28:

[0293] Compound 31 was reacted with 1,1-diphenyl-2-bromoethane using a method similar to that in Example 2 to give compound 27 below:

[0294] Yield: 26.9%

[0295] 1 H NMR(DMSO-d6)δ7.75(s,1H),7.27(m,7H),7.09(d,4H),6.91(s,2H),6.75(m,4H),6.61(d,2H),6.43(s,1H),5 .70(s,1H),5.57(t,1H),3.67(s,3H),3.60(s,3H),3.49(s,3H),3.39(q,2H),2.57(t,2H).HRMS(ESI+)calcd for C 40 H 40 NO6 + (M+H + )630.2856,found630.2848.

[0296] Example 29:

[0297] Using a method similar to that in Example 2, compound 31 was reacted with 4-methanesulfonylbenzyl bromide to give compound 28 below:

[0298] Yield: 82.0%

[0299] 1 H NMR(Chloroform-d)δ7.97(d,2H),7.76(s,1H),7.63(d,2H),7.00(d,2H),6.90(m,2H),6.81(d,2H),6.73(m,2H),6.63(dt, 1H),6.44(s,1H),5.70(s,1H),5.57(t,1H),5.13(s,2H),3.69(s,3H),3.62(s,3H),3.50(m,5H),3.06(s,3H),2.71(t,2H).

[0300] HRMS(ESI+)calcd for C 34 H 36 NO8S + (M+H + )618.2162,found 618.2157.

[0301] Example 30:

[0302] Using a method similar to that in Example 2, compound 31 was reacted with benzyl bromide to obtain compound 29 as shown below:

[0303]

[0304] Yield: 69.7%

[0305] 1 H NMR(Chloroform-d)δ7.80(s,1H),7.49–7.41(m,4H),7.40–7.35(m,1H),7.02–6.98(m,2H),6.94(dd,1H),6.92–6.85(m,3H),6.79–6. 76(m,2H),6.67(d,1H),6.49(s,1H),5.70(s,1H),5.60(t,1H),5.08(s,2H),3.74(s,3H),3.63(s,3H),3.58–3.52(m,5H),2.75(t,2H).

[0306] HRMS(ESI+)calcd for C 33 H 34 NO6 + (M+H + )540.2386, found 540.2371.

[0307] Example 31:

[0308] Using a method similar to that in Example 2, compound 31 was reacted with 4-fluorobenzyl bromide to give compound 30 as follows:

[0309]

[0310] Yield: 68.4%

[0311] 1 H NMR(DMSO-d6)δ9.27(s,1H),7.48(dt,2H),7.41(s,1H),7.21(t,2H),7.05(m,3H),6.96(dd,1H),6.89(d,2H),6.72(t,1H ),6.62(s,2H),6.56(d,1H),6.41(s,1H),5.04(s,2H),3.66(s,3H),3.60(s,3H),3.36(s,3H),3.29(m,2H),2.63(t,2H).

[0312] HRMS(ESI+)calcd for C 33 H 33 FNO6+ (M+H + )558.2292,found 558.2288.

[0313] Step C:

[0314] Example 32:

[0315] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0316] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 85.77 mg of the E-configuration compound, with a yield of 46.52%.

[0317] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.27 mmol of 4-(2-aminoethyl)phenol, 0.27 mmol of HATU, and 0.66 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 95.79 mg of the target product, with a yield of 86.12%.

[0318]

[0319] 1 H NMR(DMSO-d6)δ9.11(s,1H),7.26(t,1H),7.12(s,1H),7.03(d,1H),6.98(d,1H),6.94(d,1H),6.88(d,1H) ,6.78(m,3H),6.60(d,2H),3.78(s,3H),3.71(d,6H),2.98(q,2H),2.24(m,3H),2.21(d,2H),1.85(s,3H).

[0320] HRMS(ESI+)calcd for C 29 H 32 NO7 + (M+H + )506.2179, found 506.2167

[0321] Example 33:

[0322] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0323] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0324] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 4-(2-aminoethyl)phenol, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 87.22 mg of the target product, with a yield of 78.42%.

[0325]

[0326] 1 H NMR(400MHz,DMSO-d6)δ9.15(s,1H),7.56(t,1H),6.96(d,2H),6.86(d,1H),6.76(d ,1H),6.68(m,5H),6.45(d,1H),3.50(m,9H),3.29(m,2H),2.63(t,2H),2.18(d,6H).

[0327] HRMS(ESI+)calcd for C 29 H 32 NO7 + (M+H + )506.2179, found 506.2155

[0328] Example 34:

[0329] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0330] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 85.77 mg of the E-configuration compound, with a yield of 46.52%.

[0331] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.27 mmol of 2-(4-(prop-2-yn-1-yloxy)phenyl)ethane-1-amine, 0.27 mmol of HATU, and 0.66 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 88.96 mg of the target product, with a yield of 80.00%.

[0332]

[0333] 1 H NMR(DMSO-d6)δ7.30(t,1H),7.12(s,1H),7.04(d,1H),6.95(m,5H),6.81(m,3H),4.73(s,2H) ,3.78(s,3H),3.71(d,6H),3.54(s,1H),3.02(q,2H),2.30(t,2H),2.25(s,3H),1.86(s,3H).

[0334] HRMS(ESI+)calcd for C 32 H 34 NO7 + (M+H + )544.2335,found 544.2332

[0335] Example 35:

[0336] 1 mmol of compound 33 and 3 mmol of potassium carbonate were added to 10 mL of a methanol:water mixture (9:1) and stirred at room temperature for 1 hour. The methanol was evaporated to dryness, and 15 mL of ethyl acetate and 15 mL of water were added. The mixture was extracted and separated. The organic phase was washed once with saturated brine and dried over anhydrous sodium sulfate. 423.23 mg of the target product was obtained, with a yield of 91.30%.

[0337]

[0338] 1 H NMR(400MHz,DMSO-d6)δ9.12(s,1H),8.98(s,1H),7.13(t,1H),6.96(m,2H),6.86(dd,1H),6.81(d ,1H),6.75(m,4H),6.60(d,2H),3.76(s,3H),3.70(d,6H),3.00(q,2H),2.25(t,2H),1.81(s,3H).

[0339] HRMS(ESI+)calcd for C 27 H 30 NO6 + (M+H + )464.2073,found 464.2061

[0340] Example 36:

[0341] 1 mmol of compound 34 and 3 mmol of potassium carbonate were added to 10 mL of a methanol:water mixture (9:1) and stirred at room temperature for 1 hour. The methanol was evaporated to dryness, and 15 mL of ethyl acetate and 15 mL of water were added. The mixture was extracted and separated. The organic phase was washed once with saturated brine and dried over anhydrous sodium sulfate. 431.12 mg of the target product was obtained, with a yield of 93.07%.

[0342]

[0343] 1H NMR(DMSO-d6)δ9.14(s,1H),8.84(s,1H),7.41(t,1H),6.94(d,2H),6.78(d,1H),6.68(dd,1H),6.64(d,2H),6.54(m,2H ),6.48(s,1H),6.37(d,1H),3.55(s,3H),3.51(s,3H),3.44(s,3H),3.31(s,3H),3.27(q,2H),2.61(t,2H),2.13(s,3H).

[0344] HRMS(ESI+)calcd for C 27 H 30 NO6 + (M+H + )464.2073,found 464.2057

[0345] Example 37:

[0346] 1 mmol of compound 35 and 3 mmol of potassium carbonate were added to 10 mL of a methanol:water mixture (9:1) and stirred at room temperature for 1 hour. The methanol was evaporated to dryness, and 15 mL of ethyl acetate and 15 mL of water were added. The mixture was extracted and separated. The organic phase was washed once with saturated brine and dried over anhydrous sodium sulfate. 483.98 mg of the target product was obtained, with a yield of 96.49%.

[0347]

[0348] 1 H NMR(DMSO-d6)δ9.00(s,1H),7.19(t,1H),6.96(d,2H),6.82(m,8H),4.73(s,2H) ,3.76(s,3H),3.70(d,6H),3.54(s,1H),3.05(q,2H),2.33(t,2H),1.81(s,3H).

[0349] HRMS(ESI+)calcd for C 30 H 32 NO6 + (M+H + 502.2230, found 502.2216

[0350] Example 38:

[0351] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0352] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0353] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 2-(4-(prop-2-yn-1-yloxy)phenyl)ethane-1-amine, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 87.22 mg of the target product, with a yield of 72.91%.

[0354]

[0355] 1 H NMR(400MHz,DMSO-d6)δ7.58(t,1H),7.10(d,2H),6.86(t,3H),6.77(d,1H),6.68(m,3H),6 .45(s,1H),4.74(s,2H),3.55(s,1H),3.50(m,9H),3.33(q,2H),2.71(s,2H),2.17(d,6H).

[0356] HRMS(ESI+)calcd for C 32 H 34 NO7 + (M+H + )544.2335,found 544.2338

[0357] Example 39:

[0358] 1 mmol of compound 39 and 3 mmol of potassium carbonate were added to 10 mL of a methanol:water mixture (9:1), and the mixture was stirred at room temperature for 1 hour. The methanol was evaporated to dryness, and 15 mL of ethyl acetate and 15 mL of water were added. The mixture was extracted and separated. The organic phase was washed once with saturated brine and dried over anhydrous sodium sulfate. 455.64 mg of the target product was obtained, with a yield of 90.91%.

[0359]

[0360] 1 H NMR(DMSO-d6)δ8.85(s,1H),7.45(t,1H),7.09(d,2H),6.87(d,2H),6.79(d,1H),6.69(dd,1H),6.54(q,2H),6.48 (s,1H),6.36(d,1H),4.75(s,2H),3.55(s,3H),3.51(s,3H),3.44(s,3H),3.32(m,3H),2.68(m,2H),2.12(s,3H).

[0361] HRMS(ESI+)calcd for C 30 H 32 NO6 + (M+H + 502.2230, found 502.2214

[0362] Example 40:

[0363] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0364] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0365] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol HATU and 0.62 mmol N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water, once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 41.57 mg of the target product, with a yield of 34.75%.

[0366]

[0367] 1 H NMR(Chloroform-d)δ8.73(d,1H),8.40(d,1H),7.42(m,1H),6.94(dd,1H),6.83(d,2H),6.76(dt ,1H),6.64(s,1H),6.43(s,1H),3.97(s,3H),3.55(s,3H),3.50(s,3H),2.67(s,3H),2.28(s,3H).

[0368] 13 C NMR(Chloroform-d)δ168.89,164.29,156.18,153.40,152.29,151.67,150.69,145.63,141.04,140.43,139.70 ,135.16,129.45,122.71,120.78,120.47,117.68,115.71,113.03,111.48,56.50,55.91,55.83,23.71,20.75.

[0369] HRMS(ESI+)calcd for C 26 H 25 N4O6 + (M+H + )489.1774, found 489.1762

[0370] Example 41:

[0371] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0372] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0373] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 2-(4-((4-fluorobenzyl)oxy)phenyl)ethane-1-amine, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 129.24 mg of the target product, with a yield of 92.33%.

[0374]

[0375] 1 H NMR(Chloroform-d)δ7.40(dd,2H),7.07(t,2H),6.96(d,2H),6.81(d,2H),6.66(m,4H),6.46(dd,2H),5.9 4(t,1H),5.44(s,1H),4.98(s,2H),3.56(m,9H),3.50(q,2H),2.70(t,2H),2.35(s,3H).HRMS(ESI+)calcd for C 34 H 35 FNO6 + (M+H + )572.2448,found572.2440

[0376] Example 42:

[0377] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0378] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0379] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 2-(4-((3,5-dichlorobenzyl)oxy)phenyl)ethane-1-amine, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 142.23 mg of the target product, with a yield of 93.47%.

[0380]

[0381] 1 H NMR(Chloroform-d)δ7.40(dd,2H),7.07(t,2H),6.96(d,2H),6.81(d,2H),6.66(m,4H),6.46(dd,2H),5.9 4(t,1H),5.44(s,1H),4.98(s,2H),3.56(m,9H),3.50(q,2H),2.70(t,2H),2.35(s,3H).HRMS(ESI+)calcd for C 34 H 34 Cl2NO6 + (M+H + )622.1763,found622.1779

[0382] Example 43:

[0383] 1.24 mmol of methyl butylene, 1.86 mmol of 2-iodo-1,4-dimethoxybenzene, 2.48 mmol of (4-acetoxy-3-methoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of dicyanophenyl palladium dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The organic phases were combined, washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 193.45 mg of the isomer mixture (38.96%).

[0384] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 74.28 mg of the Z-configuration compound, with a yield of 40.29%.

[0385] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.24 mmol of 4-(2-aminoethyl)phenol, 0.24 mmol of HATU, and 0.60 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 145.54 mg of the target product, with a yield of 95.65%.

[0386]

[0387] 1 H NMR(DMSO-d6)δ9.15(s,1H),8.80(s,1H),8.04(t,1H),7.01(d,2H),6.88(d,1H),6.69(m,3H),6.50(d,1H) ,6.41(d,2H),6.35(d,1H),3.71(s,3H),3.53(s,3H),3.38(s,3H),3.34(m,2H),2.69(t,2H),1.94(s,3H).

[0388] HRMS(ESI+)calcd for C 27 H 30 NO6 + (M+H + )464.2073,found 464.2056

[0389] Example 44:

[0390] 1.24 mmol of methyl methacrylate, 1.86 mmol of 4-iodo-2-methoxy-1-((2-methoxyethoxy)methoxy)benzene, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of dicyanophenyl palladium dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The organic phases were combined, washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 227.32 mg of the isomer mixture (36.41%).

[0391] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 92.44 mg of the Z-configuration compound, with a yield of 42.48%.

[0392] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.24 mmol of 4-(2-aminoethyl)phenol, 0.24 mmol of HATU, and 0.60 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 101.37 mg of the target product, with a yield of 91.88%.

[0393]

[0394] 1 H NMR(400MHz,DMSO-d6)δ9.14(s,1H),7.47(t,1H),6.94(d,2H),6.86(d,1H),6.78(d,1H),6.69(dd,1H),6.64(d,3H),6.56(d,1H), 6.40(d,1H),5.10(s,2H),3.66(t,2H),3.55(s,3H),3.51(s,3H),3.46(s,3H),3.41(t,2H),3.29(s,5H),2.62(t,2H),2.14(s,3H).

[0395] HRMS(ESI+)calcd for C 31 H 38 NO8+ (M+H + )552.2597,found 552.2591

[0396] Example 45:

[0397] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0398] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0399] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of tyrosine methyl ester, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 95.10 mg of the target product, with a yield of 87.67%.

[0400]

[0401] 1 H NMR(Chloroform-d)δ6.68(m,8H),6.59(m,2H),6.48(d,1H),6.45(d,1H),5.45(s,1 H),4.90(dt,1H),3.70(s,3H),3.62(s,3H),3.56(d,6H),2.97(m,2H),2.31(s,3H).

[0402] HRMS(ESI+)calcd for C 29 H 32 NO8 + (M+H+ )522.2128,found 522.2137

[0403] Example 46:

[0404] 1.50 mmol of methyl methacrylate, 2.25 mmol of iodobenzene, 3.00 mmol of 2-thiopheneboronic acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of palladium dicyanophthalate were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 134.02 mg of the isomer mixture (34.60%).

[0405] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 65.30 mg of the target product, with a yield of 51.40%.

[0406] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.32 mmol of 4-(2-aminoethyl)phenol, 0.32 mmol of HATU, and 0.80 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 72.2 mg of the target product, with a yield of 73.57%.

[0407]

[0408] 1 H NMR(Chloroform-d)δ7.30(m,3H),7.16(dd,2H),7.10(dd,1H),6.86(m,2H),6.79(dd,1H),6 .76(dd,1H),6.69(m,2H),5.49(t,1H),5.40(s,1H),3.52(q,2H),2.69(t,2H),2.39(s,3H).

[0409] HRMS(ESI+)calcd for C 22 H 22 NO2S + (M+H +)364.1371,found 364.1382

[0410] Example 47:

[0411] 1.50 mmol of methyl methacrylate, 2.25 mmol of iodobenzene, 3.00 mmol of 2-thiopheneboronic acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of palladium dicyanophthalate were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 134.02 mg of the isomer mixture (34.60%).

[0412] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 35.21 mg of the target product, with a yield of 27.71%.

[0413] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.32 mmol of 4-(2-aminoethyl)phenol, 0.32 mmol of HATU, and 0.80 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 17.6 mg of the target product, with a yield of 32.28%.

[0414]

[0415] 1 H NMR(400MHz,Chloroform-d)δ7.28(m,2H),7.25(m,1H),7.14(dd,2H),7.02(m,3H),6.76 (m,3H),6.67(dd,1H),5.74(t,1H),5.21(s,1H),3.64(q,2H),2.81(t,2H),2.21(s,3H).

[0416] HRMS(ESI+)calcd for C 22 H 22 NO2S + (M+H + )364.1371,found 364.1789

[0417] Example 48:

[0418] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0419] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0420] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 2-(4-methoxyphenyl)ethane-1-amine, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 86.91 mg of the target product, with a yield of 87.97%.

[0421]

[0422] 1 H NMR(400MHz,Chloroform-d)δ6.95(m,2H),6.76(m,2H),6.66(m,4H),6.46(d,2H),5.93(t,1H ),5.44(s,1H),3.77(s,3H),3.59(s,3H),3.56(d,6H),3.50(q,2H),2.70(t,2H),2.35(s,3H).

[0423] HRMS(ESI+)calcd for C 28 H 32 NO6 + (M+H +)478.2230, found 478.2213

[0424] Example 49:

[0425] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0426] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0427] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 2-(3-methoxyphenyl)ethane-1-amine, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 82.33 mg of the target product, with a yield of 83.34%.

[0428]

[0429] 1 H NMR(400MHz,Chloroform-d)δ7.14(t,1H),6.73(dd,1H),6.69(d,1H),6.64(m,5H),6.46(d,2H ),5.98(t,1H),5.44(s,1H),3.76(s,3H),3.58(s,3H),3.56(m,6H),2.75(t,2H),2.36(s,3H).

[0430] HRMS(ESI+)calcd for C 28 H 32 NO6 + (M+H+ )478.2230, found 478.2219

[0431] Example 50:

[0432] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0433] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0434] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 2-(2-chlorophenyl)ethane-1-amine, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 83.11 mg of the target product, with a yield of 83.45%.

[0435]

[0436] 1 H NMR(400MHz,Chloroform-d)δ7.30(dd,1H),7.08(m,3H),6.66(m,4H),6.46(dd,2 H),5.98(t,1H),5.47(s,1H),3.60(s,3H),3.56(m,8H),2.92(t,2H),2.36(s,3H).

[0437] HRMS(ESI+)calcd for C 27 H 29 ClNO5 + (M+H +)482.1734, found 482.1731

[0438] Example 51:

[0439] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0440] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0441] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of (3-bromopropyl)benzene and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 94.73 mg of the target product, with a yield of 99.02%.

[0442]

[0443] 1 H NMR(400MHz,Chloroform-d)δ7.24(d,2H),7.18(m,1H),7.07(d,2H),6.71(m,4H),6.46(d,1H),6.35(d,1H) ,5.49(s,1H),4.16(t,2H),3.70(s,3H),3.55(s,3H),3.50(s,3H),2.53(m,2H),2.48(s,3H),1.91(dt,2H).

[0444] HRMS(ESI+)calcd for C 28 H 31 O6 + (M+H+ )463.2121,found 463.2114

[0445] Example 52:

[0446] 1.24 mmol of methyl butylene, 1.86 mmol of 4-iodo-2-methoxyphenyl acetate, 2.48 mmol of (2,5-dimethoxyphenyl)boric acid, 2.48 mmol of potassium carbonate, and 0.13 mmol of palladium dicyanophenyl dichloride were added to 25 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 191.08 mg of the isomer mixture (34.50%).

[0447] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 81.65 mg of the Z-configuration compound, with a yield of 44.29%.

[0448] The product obtained in the previous step was dissolved in 4 mL of dichloromethane, and 0.25 mmol of 3-methylbutane-1-amine, 0.25 mmol of HATU, and 0.62 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 76.66 mg of the target product, with a yield of 89.61%.

[0449]

[0450] 1 H NMR(400MHz,Chloroform-d)δ6.67(m,4H),6.48(dd,2H),5.86(t,1H),5.48(s,1H),3.71 (s,3H),3.56(s,6H),3.29(q,1H),2.37(s,3H),1.55(dp,1H),1.35(q,2H),0.88(d,6H).

[0451] HRMS(ESI+)calcd for C 24 H 32 NO5 + (M+H +)414.2280, found 414.2273

[0452] Example 53:

[0453] 1.50 mmol of methyl butylene, 2.25 mmol of 2-chloroiodobenzene, 3.00 mmol of (3,4-dimethoxyphenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The organic phases were combined, washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 84.01 mg of the isomer mixture (16.15%).

[0454] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water, once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 34.32 mg of the target product, with a yield of 42.97%.

[0455] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.13 mmol of 4-(2-aminoethyl)phenol, 0.13 mmol of HATU, and 0.31 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 10.68 mg of the target product, with a yield of 23.17%.

[0456]

[0457] 1 H NMR(400MHz,Chloroform-d)δ7.28(d,1H),7.04(dt,2H),6.92(d,2H),6.89(d,1H),6.69(d,2H),6.61(s,2H), 6.50(s,1H),5.53(t,1H),5.12(s,1H),3.79(s,3H),3.58(m,5H),2.74(t,2H),2.28(s,3H).HRMS(ESI+)calcd for C 26 H 27 ClNO4 + (M+H +)452.1629,found452.1641

[0458] Example 54:

[0459] 1.50 mmol of methyl methacrylate, 2.25 mmol of 2-iodo-1,4-dimethoxybenzene, 3.00 mmol of (2,5-dimethoxyphenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 169.2 mg of the isomer mixture (30.29% yield).

[0460] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water, once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 65.41 mg of the target product, with a yield of 36.50%.

[0461] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.23 mmol of 4-(2-aminoethyl)phenol, 0.23 mmol of HATU, and 0.56 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 53.75 mg of the target product, with a yield of 60.84%.

[0462]

[0463] 1 H NMR(400MHz,Chloroform-d)δ6.88(d,2H),6.69(d,2H),6.63(m,3H),6.58(dd,1H),6.52(d,1H),6.41(d,1H ),5.93(t,1H),5.40(s,1H),3.72(s,3H),3.62(s,3H),3.54(d,6H),3.49(q,2H),2.67(t,2H),2.34(s,3H).

[0464] HRMS(ESI+)calcd for C 28 H 32 NO6+ (M+H + )478.2230,found 478.2241

[0465] Example 55:

[0466] 1.50 mmol of methyl methacrylate, 2.25 mmol of 4-isopropyliodobenzene, 3.00 mmol of (2,5-dimethoxyphenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 162.60 mg of the isomer mixture (30.58% yield).

[0467] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water, once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 25.8 mg of the target product, with a yield of 16.41%.

[0468] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.10 mmol of 4-(2-aminoethyl)phenol, 0.10 mmol of HATU, and 0.24 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 15.06 mg of the target product, with a yield of 42.01%.

[0469]

[0470] 1 H NMR(400MHz,Chloroform-d)δ6.91(m,6H),6.68(m,4H),6.41(d,1H),5.93(t,1H),5.02 (s,1H),3.58(s,3H),3.50(m,5H),2.76(p,1H),2.69(t,2H),2.36(s,3H),1.14(d,6H).

[0471] HRMS(ESI+)calcd for C 29 H 34 NO4 + (M+H+ )460.2488,found 460.2493

[0472] Example 56:

[0473] 1.50 mmol of methyl methacrylate, 2.25 mmol of iodobenzene, 3.00 mmol of (2-chlorophenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 97.80 mg of the isomer mixture (22.74%).

[0474] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water, once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 65.58 mg of the target product, with a yield of 70.31%.

[0475] The product obtained in the previous step was dissolved in 5 mL of dichloromethane, and 0.24 mmol of 4-(2-aminoethyl)phenol, 0.24 mmol of HATU, and 0.72 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 60.78 mg of the target product, with a yield of 64.62%.

[0476]

[0477] 1 H NMR (400MHz, Chloroform-d) δ7.25(m,1H),7.04(m,7H),6.91(d,2H),6.87(dd,1H),6.70(d,1H),5.53(t,1H),3.55(dt,2H),2.72(t,2H),2.29(s,3H).

[0478] HRMS(ESI+)calcd for C 29 H 34 NO4 + (M+H + )392.1417, found 392.1434

[0479] Example 57:

[0480] 1.50 mmol of methyl butyl acrylate, 2.25 mmol of iodobenzene, 3.00 mmol of (4-nitrophenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 262.20 mg of the isomer mixture (58.79%).

[0481] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 113.94 mg of the target product, with a yield of 45.60%.

[0482] The product obtained in the previous step was dissolved in 5 mL of dichloromethane, and 0.49 mmol of 4-(2-aminoethyl)phenol, 0.49 mmol of HATU, and 1.21 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 90.60 mg of the target product, with a yield of 55.85%.

[0483]

[0484] 1 H NMR(400MHz,DMSO-d6)δ9.16(s,1H),8.38(t,1H),8.04(d,2H),7.32(d,2H),7.13(m, 3H),7.00(d,2H),6.94(dd,2H),6.66(d,2H),3.37(q,2H),2.69(t,2H),2.08(s,3H).

[0485] HRMS(ESI+)calcd for C 24 H 23 N2O4 + (M+H + 403.1658, found 403.1664

[0486] Example 58:

[0487] 1.50 mmol of methyl butyl acrylate, 2.25 mmol of iodobenzene, 3.00 mmol of (4-nitrophenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 262.20 mg of the isomer mixture (58.79%).

[0488] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 21.84 mg of the target product, with a yield of 8.74%.

[0489] The product obtained in the previous step was dissolved in 5 mL of dichloromethane, and 0.08 mmol of 4-(2-aminoethyl)phenol, 0.08 mmol of HATU, and 1.21 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 16.74 mg of the target product, with a yield of 53.32%.

[0490]

[0491] 1 H NMR(400MHz,DMSO-d6)δ9.12(s,1H),8.21(d,2H),7.97(t,1H),7.65(d,2H),7.42(m ,2H),7.35(m,3H),6.72(d,2H),6.57(d,2H),2.98(q,1H),2.22(t,2H),2.00(s,3H).

[0492] HRMS(ESI+)calcd for C 24 H 23 N2O4 + (M+H + 403.1658, found 403.1668

[0493] Example 59:

[0494] 1.50 mmol of methyl methacrylate, 2.25 mmol of iodobenzene, 3.00 mmol of (4-nitrophenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 180.01 mg of the isomer mixture (38.42%).

[0495] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 53.72 mg of the target product, with a yield of 31.25%.

[0496] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.22 mmol of 4-(2-aminoethyl)phenol, 0.22 mmol of HATU, and 0.54 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The product was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 31.32 mg of the target product, with a yield of 41.68%.

[0497]

[0498] 1 H NMR(400MHz,Chloroform-d)δ7.13(m,3H),7.03(d,2H),6.92(d,2H),6.70(d,2H),6.62(d,1H),6.55(dd,1H ),6.39(s,1H),5.54(d,1H),5.45(s,1H),3.81(s,3H),3.57(q,2H),3.49(s,3H),2.74(t,2H),2.30(s,3H).

[0499] HRMS(ESI+)calcd for C 26 H 28 NO4 + (M+H + )418.2018,found 418.2012

[0500] Example 60:

[0501] 1.50 mmol of methyl methacrylate, 2.25 mmol of iodobenzene, 3.00 mmol of (4-nitrophenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 180.01 mg of the isomer mixture (38.42%).

[0502] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 55.86 mg of the target product, with a yield of 32.51%.

[0503] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.22 mmol of 4-(2-aminoethyl)phenol, 0.22 mmol of HATU, and 0.54 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 31.38 mg of the target product, with a yield of 24.25%.

[0504]

[0505] 1 H NMR(400MHz,Chloroform-d)δ7.12(dd,3H),6.99(dd,2H),6.89(d,2H),6.70(m,4H),6.4 1(s,1H),5.49(t,1H),3.81(s,3H),3.55(q,2H),3.48(s,3H),2.72(t,2H),2.30(s,3H).

[0506] HRMS(ESI+)calcd for C 26 H 28 NO4 + (M+H + )418.2018,found 418.2015

[0507] Example 61:

[0508] 1.50 mmol of methyl methacrylate, 2.25 mmol of iodobenzene, 3.00 mmol of (4-nitrophenyl)boric acid, 3.00 mmol of potassium carbonate, and 0.037 mmol of dicyanophenyl palladium dichloride were added to 30 mL of a 4:1 mixture of N,N-dimethylformamide and water. The mixture was stirred at room temperature for 36 hours under argon protection. Stirring was then stopped, and the reaction mixture was poured into 100 mL of water and extracted with 80 mL of ethyl acetate, repeated three times. The combined organic phases were washed with 150 mL of saturated brine, repeated three times, and dried over anhydrous sodium sulfate. Column chromatography was used to purify the mixture, yielding 180.01 mg of the isomer mixture (38.42%).

[0509] The product obtained in the previous step was dissolved in 1 mL of tetrahydrofuran, and 10 mL of 10% NaOH aqueous solution was added. The mixture was refluxed at 110 °C for 24 h. After cooling to room temperature, the pH was adjusted to less than 3 with 2 M HCl. Extraction was performed with 10 mL of ethyl acetate, repeated three times. The organic phases were combined, washed once with water and once with saturated brine, and dried over anhydrous sodium sulfate. Purification by column chromatography yielded 55.86 mg of the target product, with a yield of 32.51%.

[0510] The product obtained in the previous step was dissolved in 3 mL of dichloromethane, and 0.22 mmol of 4-(2-aminoethyl)phenol, 0.22 mmol of HATU, and 0.54 mmol of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 2 hours. The solution was washed three times with water and once with saturated brine, and dried over anhydrous sodium sulfate. The product was purified by column chromatography to obtain 9.12 mg of the target product, with a yield of 11.80%.

[0511]

[0512] 1 H NMR(700MHz,DMSO-d6)δ9.17(s,1H),7.97(t,1H),7.05(d,2H),7.01(d,2H),6.96(d,1H),6 .68(dd,2H),6.18(d,1H),3.79(s,3H),3.77(s,3H),3.28(q,1H),2.63(t,2H),2.45(s,3H).

[0513] HRMS(ESI+)calcd for C 20 H 24 NO4 + (M+H + )342.1705, found 342.1693

[0514] Examples of cell and animal experiments:

[0515] Experimental Example 1

[0516] Hydrogen peroxide-damaged SH-SY5Y cell drug screening method

[0517] 1SH-SY5Y neural cell culture method

[0518] SH-SY5Y neural cells were cultured in DMEM medium (containing 5% fetal bovine serum) at 37°C in an incubator containing 5% CO2, and passaged every 3–4 days. Cells in the logarithmic growth phase were selected for experiments.

[0519] 2. Hydrogen peroxide model screening method

[0520] SH-SY5Y cells were used at a rate of 5 × 10⁻⁶ 3 The drug was inoculated into 96-well plates at a concentration of [specific concentration not specified] and cultured for 24 h. Then, 100 μL of H2O2-containing culture medium was added to each well to bring the final H2O2 concentration to 400 μM. The final drug concentrations were 10 μM, 1 μM, and 0.1 μM, with three parallel wells for each concentration. The plates were cultured for another 24 h. After 24 h, the supernatant was discarded, and 100 μL of MTT (0.5 mg / mL) was added to each well. The plates were incubated for another 4 h, and the supernatant was discarded again. Then, 150 μL of DMSO was added to each well, and the plates were shaken for 10 min. The absorbance was measured at 570 nm using a microplate reader.

[0521] (Efficacy rate % = (OD drug - OD model) / (OD control - OD model) * 100)

[0522] 3 Experimental Results

[0523] Protective effect of candidate compounds on SH-SY5Y cells

[0524] The results are shown in Table 1 and... Figure 1 The cytoprotective effects of the compounds were quantitatively analyzed using absorbance values. Compound 12 exhibited significant cytoprotective effects compared to other compounds, with results showing a marked improvement over fenopromine (Fenle). * (P < 0.05). Compound 12 was identified as the next experimental compound through cell screening.

[0525] Table 1. Protective effects of compounds on SH-SY5Y cells in the H2O2 damage model.

[0526]

[0527] * P < 0.05, compared with the fenoproline group.

[0528] Experiment Example 2

[0529] Determination of intracellular lanosterol content

[0530] 1. Cell Culture and Intracellular Lanosterol Assay Methods

[0531] SH-SY5Y cells were placed in 10cm culture dishes containing DMEM medium with 10% FBS (Gibco) and incubated at 37°C for 48 hours in a 5% CO2 cell culture incubator. After incubation, the original medium was removed and the cell dishes were washed three times with PBS. Then, 20μM of drug-containing medium was added, and incubation continued for 2 hours. After incubation, the original medium was removed and the cell dishes were washed three times with PBS. 0.5mL of trypsin (Gibco) was added for digestion for 3 minutes. After digestion, 3mL of DMEM medium with 10% FBS (Gibco) was added to the cell dishes to stop digestion, and the cells were collected by pipetting into 10mL centrifuge tubes. The cells were centrifuged at 1000rpm at room temperature for 5 minutes, the supernatant was removed, and 1mL of PBS was added to collect the pellet into a 1.5mL EP tube. The cells were centrifuged again at 1000rpm at room temperature for 5 minutes, the supernatant was removed, and 800μL of methanol (AR) was added and vortexed for 10 minutes. After vortexing to homogenize, centrifuge at 14000 rpm and 4°C for 10 minutes. Collect the supernatant into another 1.5 mL EP tube, remove the solution using a vacuum centrifuge, and store in a -80°C freezer for further GC-MS analysis.

[0532] Preparation: The reference drug was fenoxaprop-P-ethyl (Acta Pharmaceutica Sinica B 2021; 11(5):1213-1226), and other compounds were prepared in the laboratory according to the aforementioned method. A certain amount of sample was weighed and added to dimethyl sulfoxide (AR) to prepare a clear stock solution with a drug concentration of 50 mM. The sample stock solution was mixed evenly with DMEM medium containing 10% FBS (Gibco) at a ratio of 2 μL: 5 mL to obtain a drug-containing medium with a sample concentration of 20 μM. The medium was prepared fresh for each use.

[0533] 2GC-MS determination

[0534] Sample pretreatment: Add 300 μL of methanol to an EP tube, vortex for 1 minute, centrifuge at 14000×g for 10 minutes, take all supernatant and dry under nitrogen, add 100 μL of acetonitrile to redissolve, then add 100 μL of N-methyltrimethylsilyltrifluoroacetamide (Macklin) and derivatize at 60℃ in the dark for 60 minutes. After the reaction, centrifuge as above and take 100 μL of supernatant. Inject 2 μL of the supernatant into the gas chromatography-mass spectrometry (GC-MS) and calculate the standard curve using the external standard method as follows: 10, 50, 100, 200, 500, 1000 ng / mL.

[0535] Gas chromatography-mass spectrometry (GC-MS): The injection port temperature was set to 280℃, and the sample was injected in splitless mode. A SH-Rxi-5-Sil MS column (30m × 0.25mm id × 0.25μm) was used for separation. Helium was used as the carrier gas. The column oven temperature was programmed as follows: 75℃ initially, held for 1 minute; increased to 150℃ at 15℃ / min; then increased to 280℃ at 40℃ / min; finally increased to 310℃ at 3℃ / min and held for 3 minutes. The ion source temperature was 200℃, and the MS was operated in electron collision mode (EI). Lanosterol was monitored in SIM mode (m / z): 393, 483, 498, and quantified at 393.

[0536] 3 Experimental Results

[0537] Effect of compound 12 on lanosterol content in SH-SY5Y cells

[0538] The results are shown in Table 2 and Figure 2 The intracellular lanosterol content was quantitatively analyzed using fragment ion 393. Compound 12 significantly increased the intracellular lanosterol content in SH-SY5Y cells, with results significantly superior to fenpropathrin (…). && (P < 0.01). Since the increase in lanosterol content contributes to neuroprotection in the brain, this result indicates that compound 12 has a significant neuroprotective effect.

[0539] Table 2 shows the effects of some compounds on lanosterol content in SH-SY5Y cells.

[0540]

[0541] ** P < 0.01, *** P < 0.001, compared with the DMSO group; & P < 0.05, compared with the fenoproline group.

[0542] Experimental Example 3

[0543] Determination of lanosterol content in the brain

[0544] 1. Animal administration method and extraction method of lanosterol from the brain

[0545] C57 / BL6J mice were acclimatized for three days, then randomly divided into a blank control group, an MPTP model group, a compound 12 group (75 mg / kg), and a fenoxaprop-P-ethyl group (75 mg / kg), with 8 mice in each group. Mice in each group were administered the corresponding drug dose by gavage at 10 mL / kg, followed by an intraperitoneal injection of MPTP (30 mg / kg) 30 minutes later, once daily for 7 consecutive days. Half an hour after the last administration, the animals were processed, and the midbrain of each mouse was harvested and stored at -80°C for later use. Brain tissue samples were homogenized with physiological saline at a mass:volume ratio of 1:2. 50 μL of the homogenate was added to 50 μL of purified water and 5 μL of methanol solution containing a 400 ng / mL carbamazepine internal standard. The mixture was vortexed for 30 seconds, then extracted with 500 μL of ethyl acetate and vortexed for 3 minutes. The mixture was centrifuged at 5000 rpm for 5 minutes, and the supernatant was collected. The supernatant was dried under nitrogen at room temperature and reconstituted with 50 μL of acetonitrile:water solution (40:60). The mixture was sonicated for 15 seconds, centrifuged at 13000 rpm for 10 minutes, and the supernatant was collected for GC-MS analysis.

[0546] Preparation: The reference drug, Fefenidine, was obtained from the literature (Acta Pharmaceutica Sinica B 2021; 11(5):1213-1226). All other compounds were prepared in the laboratory according to the aforementioned method. Weigh 150 mg of the corresponding sample at a time, dissolve it in 0.4 mL of DMSO, and after complete dissolution, add 10 mL of PEG300, mix well, add 60 μL of Tween 80, mix well, add 9.54 mL of 0.5% CMC-Na solution, mix well, protect from light, and prepare fresh before use.

[0547] 2GC-MS determination

[0548] All supernatant was dried under nitrogen, redissolved in 100 μL of acetonitrile, and then 100 μL of N-methyltrimethylsilyltrifluoroacetamide (Macklin) was added. The reaction was carried out at 60 °C in the dark for 60 minutes. After the reaction, the supernatant was centrifuged as above, and 100 μL of the supernatant was injected into 2 μL of the gas chromatography-mass spectrometry (GC-MS). The standard curve was calculated using the external standard method and is as follows: 10, 50, 100, 200, 500, 1000 ng / mL.

[0549] Gas chromatography-mass spectrometry (GC-MS): The injection port temperature was set to 280℃, and the sample was injected in splitless mode. A SH-Rxi-5-Sil MS column (30m × 0.25mm id × 0.25μm) was used for separation. Helium was used as the carrier gas. The column oven temperature was programmed as follows: 75℃ initially, held for 1 minute; increased to 150℃ at 15℃ / min; then increased to 280℃ at 40℃ / min; finally increased to 310℃ at 3℃ / min and held for 3 minutes. The ion source temperature was 200℃, and the MS was operated in electron collision mode (EI). Lanosterol was monitored in SIM mode (m / z): 393, 483, 498, and quantified at 393.

[0550] 3 Experimental Results

[0551] Effect of compound 12 on lanosterol content in the brain of MPTP mice

[0552] The results are shown in Table 3 and Figure 3 The content of lanosterol in the brain was quantitatively analyzed using fragment ion 393. The lanosterol content in the brain of MPTP model mice was significantly reduced, and compound 12 significantly increased the lanosterol content in MPTP mice, with results significantly superior to fenproxil fumarate (Fenrod). & (P < 0.01), however, fenproxetine failed to significantly increase the lanosterol content in the brains of MPTP mice. Since an increase in lanosterol content contributes to neuroprotection in the brain, this result indicates that compound 12 has a significant neuroprotective effect.

[0553] Table 3. Effects of compound 12 on lanosterol levels in the brains of MPTP mice (n=8)

[0554]

[0555] ## P < 0.01, compared with the blank control group; ** P < 0.01, compared with the model group, && P < 0.01 vs. mice in the fenprolene group.

[0556] Experiment Example 4

[0557] Detection of compounds using MPTP-induced PD animal models

[0558] 1. Establishment of MPTP-induced PD mouse model and drug administration method

[0559] C57 / BL6J mice were acclimatized for three days, then randomly divided into a blank control group, an MPTP model group, a compound 12 group (75 mg / kg), and a fenoxaprop-P-ethyl group (75 mg / kg), with 29-35 mice in each group. Mice in each group were administered the corresponding drug dose by gavage at 10 mL / kg. 30 minutes after administration, MPTP (30 mg / kg) was injected intraperitoneally once daily for 7 consecutive days. After MPTP injection was discontinued, mice were administered the test drug and positive control drug once daily for 5 consecutive days. On day 12, behavioral tests of stick spinning, pole climbing, and grip strength were performed. After the last behavioral test, the animals were processed. The midbrain and striatum of each group were collected and stored at -80℃ for later use. Additionally, the brains of four mice from each group were fixed in 4% paraformaldehyde.

[0560] Preparation: MPTP was provided by Sigma-Aldrich; weigh 12,150 mg of the compound and dissolve it in 0.4 mL of DMSO. After complete dissolution, add 10 mL of PEG300 and mix well. Add 60 μL of Tween 80 and mix well. Add 9.54 mL of 0.5% CMC-Na solution and mix well. Prepare a 7.5 mg / mL solution of fenolecidine with 0.5% CMC-Na. Prepare a 3 mg / mL solution of MPTP with 0.9% physiological saline. Protect from light and prepare fresh before use.

[0561] 2 detection indicators

[0562] (1) Rotating doll test: The rotating doll test is a classic method for assessing the motor coordination ability of mice. The rotating doll device (UgoBasie) consists of a horizontal metal rod (3 cm in diameter) approximately 50 cm long, divided into 5 segments by metal plates, with baffles separating the animals so they do not interfere with each other. The rotating doll device is rotated at a constant speed of 25 r / min. The mouse is then placed on the rod, and the timer is started until the mouse falls off the rod. This is recorded as the latency period (i.e., the time of the first fall), which represents its motor coordination ability. Each mouse is tested twice, with a 30-minute interval, and the average of the two results is taken.

[0563] (2) Pole Climbing Test: The pole climbing test is commonly used to evaluate limb coordination in mice. A pole climbing apparatus (Institute of Materia Medica, Chinese Academy of Medical Sciences) with a diameter of 13 mm and a height of 150 mm is used. The mouse is placed head down on top of the metal pole and allowed to climb down naturally. The animal's behavior during the descent is observed. The mouse's behavior during the descent is scored according to the following criteria:

[0564] 5 points: Using all four limbs, crawling downwards in a coordinated manner step by step;

[0565] 4 points: Crawling downwards step by step, but also exhibiting gliding behavior with hind limbs;

[0566] 3 points: After climbing halfway, slide downhill, but you can hold on to the metal pole;

[0567] 2 points: Sliding behavior occurred before climbing more than half the distance;

[0568] 1 point: Do not fall off the pole after climbing halfway;

[0569] 0 points: Failed to grab the pole before climbing halfway and fell off.

[0570] 3. Statistical Analysis

[0571] Experimental data are presented as mean ± standard error (Mean ± SEM). Differences between different groups were compared using one-way ANOVA, with P < 0.05 indicating a significant difference.

[0572] 4 Experimental Results

[0573] 4.1 Effect of compound 12 on the spinneret test in MPTP-induced PD mice

[0574] The results are shown in Table 4 and Figure 4 Motor dysfunction is a major pathological feature of Parkinson's disease (PD). To detect whether MPTP-induced model mice exhibited PD-like motor symptoms, a stick-spinning test was performed on day 12 of the experiment. The stick-spinning test is a commonly used experimental method for assessing motor coordination in rodents. The results showed that the MPTP model group mice spent significantly less time on the stick than the blank control group. Administration of compound 12 significantly increased the time spent on the stick in MPTP model mice, superior to fenproxil fumarate (…). & (P < 0.05). The results indicate that administration of the test drug compound 12 had a good effect on improving motor coordination dysfunction induced by MPTP in PD mice.

[0575] Table 4. Effects of compound 12 on the rotarod test in MPTP-induced PD mice (n = 29-35)

[0576]

[0577] ### P < 0.001, compared with the blank control group; * P < 0.05 *** P < 0.001, compared with the model group, & P < 0.05 vs. mice in the fenprolene group.

[0578] 4.2 Effect of compound 12 on MPTP-induced pole climbing test in PD mice

[0579] The results are shown in Table 5 and Figure 5The pole climbing test can be used to assess the motor function of the limbs in mice. On day 12 of the experiment, the pole climbing test was performed on mice. The results showed that the pole climbing test scores of the MPTP model group were significantly lower than those of the blank control group. Compound 12 significantly improved the pole climbing test scores of mice, comparable to that of fenoxaprop-P-ethyl. These results indicate that compound 12 has a good ameliorative effect on MPTP-induced motor dysfunction.

[0580] Table 5. Effects of compound 12 on MPTP-induced pole climbing test in PD mice (n = 29-35)

[0581]

[0582]

[0583] ### P < 0.001, compared with the blank control group; *** P < 0.001, compared with the model group.

[0584] Experiment 5: Compound Detection Using the A53T Transgenic PD Mouse Model

[0585] 1A53T transgenic PD mice and administration method

[0586] SPF-grade α-Syn(A53T) (male), 6 months old, weighing 22-24g; and SPF-grade wild-type C57BL / 6J mice (WT mice, male), 6 months old, weighing 22-24g, were purchased from Changzhou Cavens Laboratory Animal Co., Ltd. Five mice were housed in barrier-grade SPF animal enclosures at room temperature with free access to food and water. All procedures performed in accordance with the regulations of the Beijing Laboratory Animal Ethics Committee.

[0587] Six-month-old α-Syn(A53T) and WT mice underwent monthly behavioral testing. At 12 months of age, A53T transgenic mice exhibited behavioral impairments. Based on the behavioral results, A53T mice were divided into four groups: the A53T model group, the compound 12 group (75 mg / kg), the fenopromine group (75 mg / kg), and the positive control levodopa group (20 mg / kg), with 14-23 mice in each group. Mice in each treatment group were administered the corresponding drug dose via gavage at 10 mL / kg for 8 consecutive weeks. Mice in the WT and A53T groups were administered the corresponding solvent dose via gavage. At the second month after drug administration, behavioral tests were performed on a stick, pole, and balance beam. After the last behavioral test, the animals were processed. Blood was collected from the eyes of mice in each group, allowed to stand at room temperature for 2 hours, centrifuged at 3000 rpm for 10 minutes, and the serum was stored at -80℃ for later use. Additionally, the brains of four mice from each group were fixed in 4% paraformaldehyde, and the striatum and midbrain of the remaining mice were used for subsequent studies.

[0588] The positive control drug, levodopa tablets, was provided by Beijing Shuguang Pharmaceutical Co., Ltd.; levodopa was prepared into a 2 mg / mL solution using 0.5% CMC-Na; fenproxil was prepared into a 7.5 mg / mL solution using 0.5% CMC-Na; preparation of compound 12: 150 mg of compound 12 was weighed and dissolved in 0.4 mL of DMSO. After complete dissolution, 10 mL of PEG300 was added and mixed well. Then, 60 μL of Tween 80 was added and mixed well. Finally, 9.54 mL of 0.5% CMC-Na solution was added and mixed well.

[0589] 2. Detection indicators:

[0590] (1) Rotating doll test: The rotating doll test (Ugo Basie) uses a horizontal metal rod (3cm in diameter) approximately 50cm long, divided into 5 sections by metal plates, with baffles separating the animals so they are not affected by each other. The rotating doll test is run at a constant speed, and the mouse is placed on the rod. Timing begins, and the latency period (i.e., the time of the first fall) until the mouse falls off the rod is recorded as the latency period, which represents its motor coordination ability. Each mouse is tested twice, with a 30-minute interval, and the average of the two results is taken.

[0591] (2) Pole Climbing Test: A pole climbing device (Institute of Materia Medica, Chinese Academy of Medical Sciences) with a diameter of 13 mm and a height of 150 mm was used. The device was erected vertically, and the mouse was placed head down on the top of the metal pole, allowing it to climb down naturally. The animal's behavior during the descent was observed. The mouse's behavior during the descent was scored according to the following criteria:

[0592] 5 points: Using all four limbs, crawling downwards in a coordinated manner step by step;

[0593] 4 points: Crawling downwards step by step, but also exhibiting gliding behavior with hind limbs;

[0594] 3 points: After climbing halfway, slide downhill, but you can hold on to the metal pole;

[0595] 2 points: Sliding behavior occurred before climbing more than half the distance;

[0596] 1 point: Do not fall off the pole after climbing halfway;

[0597] 0 points: Failed to grab the pole before climbing halfway and fell off. 5. Insulin resistance index test.

[0598] (3) Balance Beam Test: The balance beam is a square wooden bar 100cm long and 1cm wide, with a black box covered with sawdust at one end. The experiment was divided into a training period and a testing period. During the training period, the mouse was placed 80cm away from the black box end of the balance beam and allowed to walk into the black box. After 30 minutes, the mouse was placed 80cm away from one end of the balance beam and allowed to walk to the black box at the other end. The time taken for the mouse to cross the balance beam from the starting position to the point where all four limbs were in the black box, as well as the number of times it slipped, were recorded. The mouse's behavior on the balance beam was also recorded and scored according to Feeney's scoring criteria, which are as follows:

[0599] A score of 0 means passing through the balance beam without falling;

[0600] A score of 1 indicates that the chance of falling while crossing the balance beam is less than 50%.

[0601] A score of 2 indicates a greater than 50% chance of falling while crossing the balance beam;

[0602] A score of 3 indicates that the person can walk across the balance beam, but the affected paralyzed hind limb cannot help move forward.

[0603] A score of 4 indicates that you cannot walk across the balance beam, but you can sit on it;

[0604] A score of 5 indicates that the mouse will fall off the balance beam.

[0605] 3. Statistical Analysis

[0606] Experimental data are presented as mean ± standard error (Mean ± SEM). Differences between different groups were compared using one-way ANOVA, with P < 0.05 indicating a significant difference.

[0607] 4 Experimental Results

[0608] 4.1 Effects of compound 12 on the transgenic behavior of A53T transgenic mice

[0609] The results are shown in Table 6 and Figure 6 Motor dysfunction is a major pathological feature of Parkinson's disease (PD). To detect whether A53T transgenic mice exhibited PD-like motor symptoms, mice in each group underwent a stick-spinning test at the end of week 8 after drug administration. The stick-spinning test is a commonly used experimental method for assessing motor coordination in rodents. The results showed that the time mice in the compound 12 group spent on the stick was significantly longer than that in the A53T group, and superior to that of fenproxil fumarate and levodopa. This indicates that compound 12 has a good effect on improving motor coordination dysfunction in A53T mice.

[0610] Table 6. Effects of compound 12 on the transposon test in A53T transgenic mice (n = 14-23)

[0611]

[0612] ### P < 0.001, compared with the wild-type group; * P < 0.05 *** P < 0.001, compared with A53T transgenic genome.

[0613] 4.2 Effects of compound 12 on pole-climbing behavior in A53T transgenic mice

[0614] The results are shown in Table 7 and Figure 7 The pole climbing test can be used to assess the motor function of the limbs in mice. Mice in each group underwent the pole climbing test at the end of week 8 after drug administration. The results showed that the scores of mice in the compound 12, fenolephrine, and levodopa groups were significantly different from those in A53T mice, indicating that compound 12 had a certain ameliorative effect on the motor dysfunction of A53T mice.

[0615] Table 7. Effects of compound 12 on pole climbing test in A53T transgenic mice (n = 14-23)

[0616]

[0617] ### P < 0.001, compared with the wild-type group; * P < 0.05 ** P < 0.01, compared with A53T transgenic genome.

[0618] 4.3 Effects of Compound 12 on Balance Behavior of A53T Transgenic Mice

[0619] The results are shown in Table 8 and Figure 8 The balance beam test is commonly used to evaluate the fine motor skills of mice, as well as the balance and coordination of their forelimbs and hindlimbs. Evaluation indicators include the time taken for the mouse to cross the beam and the number of slips. Mice in each group underwent the balance beam test at the end of week 8 after drug administration. Results showed that the A53T transgenic mice had significantly longer time on the balance beam and fewer falls than wild-type mice. The mice in the compound 12 group had significantly shorter crossing time, comparable to that of fenproxetine. & (P < 0.05); The number of falls in the compound 12 group was significantly reduced, which was better than that in the fenoxaprop-P-ethyl group and comparable to that in the levodopa group. These results indicate that compound 12 can significantly improve motor dysfunction in PD mice.

[0620] Table 8. Effects of compound 12 on the balance beam test in A53T transgenic mice (n = 14-23)

[0621]

[0622] ### P < 0.001, compared with the wild-type group; * P < 0.05 ** P < 0.01, *** P < 0.001, compared with A53T transgenic genome; & P < 0.05 vs. mice in the fenprolene group.

[0623] Example 6: Detection of the compound using a scopolamine AD mouse model

[0624] 1. Establishment and administration method of scopolamine AD mice

[0625] ICR mice were acclimatized for three days, then randomly divided into a blank control group, a scopolamine model group, a compound 12 group (75 mg / kg), a donepezil group (5 mg / kg), and a memantine group (5 mg / kg), with 11-15 mice in each group. Mice in each group were administered the corresponding dose of the drug by gavage at 10 mL / kg once daily for 7 consecutive days. One hour after drug administration on day 7, the model group and each drug administration group were intraperitoneally injected with scopolamine (2 mg / kg). Thirty minutes later, a behavioral test was performed using the jumping test. The time of the first jump (latency period) and the number of jumps within 5 minutes were recorded. After the last behavioral test, the animals were processed. Cortex and hippocampus samples from each group were stored at -80°C for later use. Additionally, the brains of four mice from each group were fixed in 4% paraformaldehyde.

[0626] Preparation: Donepezil tablets (positive control) were provided by Eisai (China) Pharmaceutical Co., Ltd., memantine by Lundbeck Pharmaceuticals, Denmark, and scopolamine by Sigma-Aldrich. Donepezil and memantine were prepared to a concentration of 5 mg / mL using 0.5% CMC-Na. 12,150 mg of the compound was weighed and dissolved in 0.4 mL of DMSO. After complete dissolution, 10 mL of PEG300 was added and mixed. 60 μL of Tween 80 was added and mixed. 9.54 mL of 0.5% CMC-Na solution was added and mixed. Scopolamine was prepared to a concentration of 0.2 mg / mL using 0.9% physiological saline. The solution was kept away from light and prepared fresh before use.

[0627] 2 detection indicators

[0628] The jumping platform experiment setup consisted of a rectangular reflective box measuring 10cm × 10cm × 60cm, divided into 5 compartments by black plastic panels. The bottom was covered with copper grids spaced 0.5cm apart. The box was electrified, with the voltage controlled by a transformer. A wooden platform, 4.5cm high and in diameter, was placed in the right corner of each compartment. During the experiment, a 36V AC power supply was applied. The normal reaction of mice after receiving an electric shock was to jump back to the safe platform to avoid the harmful stimulus. On the first day, the power was off, and the mice were allowed to move freely in the reflective box for 5 minutes to familiarize themselves with the environment. After 24 hours, the copper grid power supply (36V AC) was connected. The time from receiving the electric shock to first jumping onto the safe platform (reaction time) and the number of errors jumping off the safe platform within 5 minutes were recorded for each group of mice, serving as the learning test score. The process was repeated the following day, recording the time for the first jump off the safe platform (latency period) and the number of electric shocks received within 5 minutes (error count), serving as the memory test score. During the experiment, if the mouse remained on the safe platform for more than 5 minutes, its latency period was calculated as 5 minutes.

[0629] 3. Statistical Analysis

[0630] Experimental data are presented as mean ± standard error (Mean ± SEM). Differences between different groups were compared using one-way ANOVA, with P < 0.05 indicating a significant difference.

[0631] 4 Experimental Results

[0632] Effects of Compound 12 on the Jumping Behavior of Scopolamine AD Mice

[0633] The results are shown in Table 9 and Figure 9 The platform jumping test is commonly used to evaluate the learning and memory abilities of mice. Evaluation indicators include the time it takes for the mouse to jump off the platform for the first time (latency) and the number of electric shocks received within 5 minutes (number of errors). Results showed that the scopolamine-induced model mice had a significantly shorter platform jumping latency and a significantly higher number of errors than the control group. Compound 12 could prolong the platform jumping latency to some extent and significantly reduce the number of errors, comparable to the positive control drugs donepezil and memantine. These results indicate that compound 12 can significantly improve learning and memory impairment in AD mice.

[0634] Table 9. Effects of compound 12 on scopolamine AD mice jumping platform (n = 13-15)

[0635]

[0636] ### P < 0.001, compared with the blank control group; * P < 0.05 ** P < 0.01, compared with the model group.

Claims

1. A compound of the general formula or a pharmaceutically acceptable salt thereof or its corresponding isomer. Its features are: Ring A is a benzene ring or an aromatic heterocycle containing 3-6 carbon atoms; X can be an alkylene group with 1-5 carbon atoms or an alkylene group containing a carbonyl group with 1-5 carbon atoms; R1 and R2 can be located at any position in ring A; R1 and R2 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

2. A compound of the general formula or a pharmaceutically acceptable salt thereof or its corresponding isomer. Its features are: Ring A is a benzene ring; X can be an alkylene group with 1-5 carbon atoms or an alkylene group containing a carbonyl group with 1-5 carbon atoms; R1 and R2 can be located at any position in ring A; R1 and R2 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

3. A compound of the general formula or a pharmaceutically acceptable salt thereof or its corresponding isomer. Rings A, B, and C are benzene rings or aromatic heterocycles containing 3-6 carbon atoms; X can be 0 or N; n can be 0-5 R1 and R2 can be located at any position in ring A; R3 and R4 can be located at any position in ring B; R5 and R6 can be located at any position in ring C. R1, R2, R3, R4, R5, and R6 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen; R7 can be a straight-chain or branched alkyl group with 1-5 carbon atoms (oxygen-containing or oxygen-free).

4. A compound of the general formula or a pharmaceutically acceptable salt thereof or its corresponding isomer. Rings A, B, and C are benzene rings; X can be O or N; R1 and R2 can be located at any position in ring A; R3 and R4 can be located at any position in ring B; R5 and R6 can be located at any position in ring C. R1, R2, R3, R4, R5, and R6 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

5. The compound or its isomers according to claims 1-4, characterized in that... Including compounds of formula (Ia-1) in: Ring A is a benzene ring; R1 and R2 can be located at any position in ring A; R1 and R2 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

6. The compound or its isomers according to claims 1-4, characterized in that... Including the following compound (Ia-2) in Ring A is a benzene ring; R1 and R2 can be located at any position in ring A; R1 and R2 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

7. The compound or its isomers according to claims 1-4, characterized in that... Including compounds of formula (Ia-3) in: Ring A and ring A” are benzene rings; R1 and R2 can be located at any position in ring A; R1 and R2 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen. R1” and R2” can be located at any position in ring A; R1” and R2” are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

8. The compound or its isomers according to claims 1-4, characterized in that... Including compound (II-1) of the following formula. in: Ring A is a benzene ring or an aromatic heterocycle containing 3-6 carbon atoms; rings B and C are benzene rings. X can be O or N; R1 and R2 can be located at any position in ring A; R3 and R4 can be located at any position in ring B; R5 and R6 can be located at any position in ring C. R1, R2, R3, R4, R5, and R6 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

9. The compound or its isomers according to claims 1-4, characterized in that... Including compound (II-2) of formula below. in: Ring B is a benzene ring or an aromatic heterocycle containing 3-6 carbon atoms; rings A and C are benzene rings. X can be O or N; R1 and R2 can be located at any position in ring A; R3 and R4 can be located at any position in ring B; R5 and R6 can be located at any position in ring C. R1, R2, R3, R4, R5, and R6 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

10. The compound or its isomers according to claims 1-4, characterized in that... Including compound (II-3) in: Rings A and B are benzene rings; X can be 0 or N; n can be 0-5; R1 and R2 can be located at any position in ring A; R3 and R4 can be located at any position in ring B. R1, R2, R3, and R4 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

11. The compound or its isomers according to claims 1-4, characterized in that... Including compound (II-4) in: Rings A and C are benzene rings; X can be O or N; R1 and R2 can be located at any position in ring A; R5 and R6 can be located at any position in ring C. R1, R2, R5, and R6 are independently selected from hydrogen, straight-chain or branched alkyl groups with 1-5 carbon atoms (oxygen-containing or oxygen-free), halogen-substituted straight-chain or branched alkyl groups with 1-5 carbon atoms, nitro, cyano, acyloxy, sulfonyl, and halogen.

12. The compound or its isomer according to any one of claims 1-11, characterized in that... The compounds include:

13. A pharmaceutical composition, characterized in that... It contains the compound of any one of claims 1-12, or a pharmaceutically acceptable salt thereof, or an isomer thereof as the active ingredient and a pharmaceutically acceptable carrier.

14. The pharmaceutical composition according to claim 13, characterized in that, The pharmaceutical composition is selected from tablets, capsules, pills, or injections.

15. Use of any compound of claims 1-12, or a chemically acceptable salt thereof, or an isomer thereof, in the preparation of medicaments for the prevention and / or treatment of neurodegenerative diseases.

16. The application according to claim 15, characterized in that, The neurodegenerative disease mentioned is selected from Parkinson's disease.

17. Use of any compound of claims 1-12, or a chemically acceptable salt thereof, or an isomer thereof, in the preparation of a medicament for improving learning and memory impairment or for treating memory loss and Alzheimer's disease.

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