CN117800979A - A kind of tetrahydrobenzofuran[2,3-c]pyridine kinase inhibitor and its preparation method and use - Google Patents

Abstract

The invention relates to a type of tetrahydrobenzofuran[2,3-c]pyridine kinase inhibitor and its preparation method and use. Specifically, the compound of the present invention has the structure shown in Formula I, in which each group and substituent is defined as described in the specification. The invention also discloses the preparation method of the compound and its use in anti-tumor aspects.

Description

Tetrahydrobenzofuran [2,3-c ] pyridine kinase inhibitor and preparation method and application thereof

Technical Field

The invention relates to the field of medicines, in particular to a tetrahydrobenzofuran [2,3-c ] pyridine kinase inhibitor, a preparation method and application thereof.

Background

p 21-activated kinases (PAKs) are intracellular serine/threonine-specific protein kinases that play an important role in cytoskeletal organization, cell morphogenesis and cell survival. Is involved in the regulation of a variety of diseases including cancer, infectious diseases and neurological diseases. PAKs are located at junctions of signal pathways necessary for the development of a variety of tumors. When PAKs are over-expressed, mutated or abnormally activated by upstream signals such as the cell division controlling proteins Cdc42 or RAC, pathways such as ERK, AKT and WNT are affected, and oncogenic signals are generated in the cell, including promoting autonomous growth signals, evading apoptosis, and promoting invasion, metastasis, etc. In addition, PAKs proteins are upregulated in a variety of human cancers and are positively correlated with tumor advanced levels and patient survival decreases. Thus, the development of PAK inhibitors is an important strategy for the treatment of tumors.

The PAKs family consists of six members, which fall into two categories according to sequence and structural homology: type I (PAK 1-3) and type II (PAK 4-6). However, type I PAKs have proven to be very challenging small molecule drug targets. Moreover, a recent study indicated that PAK2 inhibition is associated with acute cardiovascular toxicity and would be enhanced by PAK1 inhibition. This study led to alarming studies with pan I-type PAKs inhibitors. While PAK4 type II promotes BRAF or KRAS driven tumor invasion, metastasis and proliferation, which is of great interest. Meanwhile, recent studies have shown that PAK4 is involved in tumor immunity. PAK4 is enriched in tumor biopsies with low T cell and dendritic cell infiltration and is inversely related to immune marker response and T cell infiltration. In addition, gene knockout of PAK4 can induce re-expression of endothelial cell adhesion protein, reduce vascular abnormalities and improve T cell infiltration. In the B16 melanoma mouse model, PAK4 knockdown would make PD-1 antibody treatment more sensitive and improve T cell infiltration.

Currently, only one PAK4 inhibitor, ATG-019 (KPT-9274), developed by Karyopharm corporation, is in phase I clinical study for the treatment of advanced solid tumors or non-Hodgkin's lymphoma. KPT-9274 is a PAK4 allosteric inhibitor, and does not directly inhibit PAK4 kinase activity, but can down regulate the phosphorylation level of PAK4 protein and downstream signals in tumor cells; moreover, KPT-9274 has no obvious inhibition effect on type I PAKs. Meanwhile, the compound has good inhibitory activity on various tumor cells and good anti-tumor activity in kidney cancer and pancreatic cancer models. Furthermore, in the mouse B16 melanoma, MC38 colon cancer and glioblastoma multiforme models, KPT-9274 in combination with PD-1 mab or CAR-T therapy improved the anti-tumor effect relative to single administration.

In conclusion, PAK4 plays an important role in tumorigenesis and development as well as tumor immune escape, so that development of small molecule drugs targeting PAK4 has good market prospect. Currently, no drug is marketed in the field, only one drug is in clinical research and development stage, and more novel PAK4 inhibitors are needed to be developed to meet clinical demands. Therefore, the inventor hopes to develop a high-selectivity PAK4 inhibitor with high efficiency, low toxicity and good metabolization, and provide a new option for tumor treatment.

Disclosure of Invention

The invention aims to provide a compound shown in a formula I, a preparation method thereof and application thereof in anti-tumor aspect.

In a first aspect of the present invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt, solvate, prodrug thereof,

wherein,

a is selected from the group consisting of:wherein R is ⁰ Selected from the group consisting of: hydrogen, C1-C6 alkyl;

v, W, X, Y are each independently selected from the group consisting of: n, CR ⁴ Wherein R is ⁴ Selected from the group consisting of: hydrogen, halogen, hydroxy, C1-C6 alkoxy, amino, cyano, C1-C6 alkyl;

z is selected from the group consisting of:

R ¹ selected from the group consisting of: hydrogen, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O, S, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O, S; wherein the substitution independently refers to substitution with one or more substituents selected from the group consisting of: halogen, hydroxy, cyano, C1-C6 alkoxy, amino, C1-C6 alkyl;

R ² Selected from the group consisting of: substituted or unsubstituted C1-C6 alkylSubstituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O, S, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O, S,Wherein n is selected from the group consisting of: 0. 1, 2, 3, said substitution independently referring to substitution with one or more substituents selected from the group consisting of: halogen, hydroxy, amino, cyano, mercapto, difluoromethyl, oxo (=o), C1-C6 alkoxy, C1-C6 alkyl, said heterocycloalkyl being selected from the group consisting of: monocyclic, parallel, spiro, bridged rings;

R ³ selected from the group consisting of: hydrogen, amino.

In another preferred embodiment, a is selected from the group consisting of:wherein R is ⁰ Selected from the group consisting of: H. C1-C6 alkyl;

v, W, X, Y are each independently selected from the group consisting of: n, CR ⁴ Wherein R is ⁴ Selected from the group consisting of: hydrogen, halogen, C1-C6 alkyl;

z is selected from the group consisting of:

R ¹ selected from the group consisting of: hydrogen, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C10 aryl; wherein the substitution independently refers to substitution with one or more substituents selected from the group consisting of: halogen, C1-C6 alkyl;

R ² Selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O, S, substituted or unsubstituted C3-C8 cycloalkyl,Wherein n is selected from the group consisting of: 0. 1, 2, saidSubstituted independently refers to substitution with one or more substituents selected from the group consisting of: halogen, hydroxy, cyano, mercapto, difluoromethyl, oxo (=o), C1-C6 alkoxy, C1-C6 alkyl, said heterocycloalkyl being selected from the group consisting of: monocyclic, parallel, spiro, bridged rings;

R ³ selected from the group consisting of: hydrogen, amino.

In another preferred embodiment, a is selected from the group consisting of:wherein R is ⁰ H.

In a further preferred embodiment of the present invention,selected from the group consisting of:

R ⁴ Selected from the group consisting of: hydrogen, halogen.

In another preferred embodiment, R ² Selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O, S, substituted or unsubstituted C3-C8 cycloalkyl,Wherein n is selected from the group consisting of: 0. 1, 2, 3, said substitution independently referring to substitution with one or more substituents selected from the group consisting of: halogen, hydroxy, cyano, mercapto, difluoromethyl, oxo (=o), C1-C6 alkoxy, C1-C6 alkyl, said heterocycloalkyl being selected from the group consisting of: monocyclic, fused, spiro, bridged rings.

In another preferred embodiment, R ² Selected from substitution orUnsubstituted groups of:C1-C6 alkyl,

In another preferred embodiment, R ¹ Selected from the group consisting of: hydrogen, halogen, substituted or unsubstituted C6-C10 aryl; wherein the substitution independently refers to substitution with one or more substituents selected from the group consisting of: halogen, C1-C6 alkyl.

In another preferred embodiment, the compound is selected from the group consisting of:

in a second aspect of the invention there is provided a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of a compound of the first aspect of the invention.

In a third aspect of the present invention there is provided the use of a compound according to the first aspect of the present invention for the manufacture of a medicament for the prevention and/or treatment of a disease selected from the group consisting of: cancer, infectious disease, neurological disease.

In a fourth aspect of the invention there is provided the use of a compound according to the first aspect of the invention for the manufacture of a medicament for inhibiting PAK4 kinase activity.

In a fifth aspect of the invention there is provided the use of a compound according to the first aspect of the invention for the manufacture of a medicament for the prophylaxis and/or treatment of diseases associated with PAK4 kinase activity.

In another preferred embodiment, the PAK4 kinase activity-related disorder is selected from the group consisting of: cancer, infectious disease, neurological disease.

In another preferred embodiment, the cancer is selected from the group consisting of: non-hodgkin's lymphoma, renal carcinoma, pancreatic carcinoma, melanoma, colon carcinoma, glioblastoma.

It is understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described below (e.g., in the examples) may be combined with each other to constitute new or preferred technical solutions. And are limited to a space, and are not described in detail herein.

Drawings

FIG. 1 is a graph of KPT-9274 (hydrochloride) solution concentration versus peak area.

FIG. 2 is a graph showing the linear relationship between the concentration of compound S21 (hydrochloride) solution and the peak area.

FIG. 3 is a graph showing the linear relationship between the concentration of compound S23 (hydrochloride) solution and the peak area.

Detailed Description

Through long-term and intensive research, the inventor unexpectedly prepares a compound with novel structure, high inhibition activity and high water solubility through structural optimization. On this basis, the inventors completed the present invention.

Terminology

In the present invention, unless otherwise indicated, terms used have the ordinary meanings known to those skilled in the art.

In the present invention, the term "halogen" refers to F, cl, br or I.

In the present invention, "C1-C6 alkyl" means a straight-chain or branched alkyl group comprising 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, neopentyl, t-pentyl, or the like.

In the present invention, the term "C2-C6 alkenyl" refers to a straight or branched alkenyl group having 2 to 6 carbon atoms containing one double bond, including without limitation ethenyl, propenyl, butenyl, isobutenyl, pentenyl, hexenyl and the like.

In the present invention, the term "C2-C6 alkynyl" refers to a straight or branched chain alkynyl group having 2 to 6 carbon atoms containing one triple bond, and includes, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentynyl, hexynyl and the like.

In the present invention, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3 to 8 carbon atoms in the ring, including, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. The term "C3-C6 cycloalkyl" has similar meaning.

In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched chain alkoxy group having 1 to 6 carbon atoms, including without limitation methoxy, ethoxy, propoxy, isopropoxy, butoxy and the like. Preferably C1-C4 alkoxy.

In the present invention, the term "heterocyclyl" is a 4-8 membered heterocyclyl containing 1, 2 or 3 heteroatoms selected from N, O, S, including, but not limited to, the following groups:

in the present invention, the term "aromatic ring" or "aryl" has the same meaning, preferably "C6-C10 aryl". The term "C6-C10 aryl" refers to an aromatic cyclic group having 6 to 10 carbon atoms, such as phenyl, naphthyl, and the like, which does not contain heteroatoms in the ring.

In the present invention, the term "aromatic heterocycle" or "heteroaryl" has the same meaning and refers to a heteroaromatic group containing one to more heteroatoms. For example, "C3-C10 heteroaryl" refers to aromatic heterocycles containing 1 to 4 heteroatoms selected from oxygen, sulfur and nitrogen, and 3 to 10 carbon atoms. Non-limiting examples include: furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring. Heteroaryl groups may be optionally substituted or unsubstituted.

In the present invention, the term "halo" refers to substitution with halogen.

In the present invention, the term "deuterated" means substituted with deuterium.

In the present invention, the term "substituted" means that one or more hydrogen atoms on a particular group are replaced with a particular substituent. The specific substituents are those described in the foregoing for each of the examples or are those found in each of the examples. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable site of the group, which may be the same or different at each position. Those skilled in the art will appreciate that combinations of substituents contemplated by the present invention are those that are stable or chemically achievable. Such as (but not limited to): halogen, hydroxy, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3-to 12-membered heterocyclyl, aryl, heteroaryl, C1-C8 aldehyde, C2-C10 acyl, C2-C10 ester, amino, C1-C6 alkoxy, C1-C10 sulfonyl, and the like.

In the present invention, the term 1-6 refers to 1, 2, 3, 4, 5 or 6. Other similar terms each independently have similar meanings. The term "plurality" refers to 2-6, such as 2, 3, 4, 5 or 6.

It will be understood that when a group is present in a compound in a plurality of different positions at the same time, the definition of each position is independent of the other and may be the same or different. That is, the term "selected from the group consisting of: the "and the term" are each independently selected from the group consisting of: "has the same meaning.

Compounds of formula (I)

The invention relates to a compound with p21 activated kinase 4 (PAK 4) inhibitory activity, a pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof, a preparation method thereof, a pharmaceutical composition containing the compound and application of the compound in preparing medicines for preventing or treating tumors, infectious diseases and neurological diseases.

The invention provides a compound which is a compound shown in a formula I or pharmaceutically acceptable salt, solvate and prodrug thereof,

wherein each group is as defined above.

In another preferred embodiment, A, V, W, X, Y, Z, R of said compounds ¹ 、R ² 、R ³ Each of which is independently a group corresponding to a particular compound described herein.

As used herein, the term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention with acids or bases that are suitable for use as medicaments. Pharmaceutically acceptable salts include inorganic and organic salts. One preferred class of salts is the salts of the compounds of the present invention with acids. Suitable salts forming acids include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, and the like; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, and the like; amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

Another preferred class of salts are salts of the compounds of the invention with bases, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (e.g., lower alkanolammonium salts and other pharmaceutically acceptable amine salts), such as methylamine, ethylamine, propylamine, dimethylamine, trimethylamine, diethylamine, triethylamine, tert-butylamine, ethylenediamine, hydroxyethylamine, dihydroxyethylamine, and triethylamine salts, and amine salts formed from morpholine, piperazine, lysine, respectively.

The term "solvate" refers to a complex of a compound of the invention coordinated to a solvent molecule to form a specific ratio. "hydrate" refers to a complex of the compound of the present invention coordinated to water.

The term "prodrug" includes a class of compounds which may themselves be biologically active or inactive, and which upon administration by an appropriate method undergo a metabolic or chemical reaction in the human body to convert to a compound of formula I, or a salt or solution of a compound of formula I. The prodrugs include, but are not limited to, carboxylic acid esters, carbonic acid esters, phosphoric acid esters, nitric acid esters, sulfuric acid esters, sulfone esters, sulfoxide esters, amino compounds, carbamates, azo compounds, phosphoramides, glucosides, ethers, acetals, and the like of the compound.

Preparation method

The following more particularly describes the preparation method of the compound of the formula I, but these specific methods do not limit the present invention. The compounds of the present invention may also be conveniently prepared by optionally combining the various synthetic methods described in this specification or known in the art, such combinations being readily apparent to those skilled in the art to which the present invention pertains.

Typically, the compounds of the present invention are prepared by the following process wherein the starting materials and reagents used, unless otherwise specified, are commercially available.

Step one:

step two:

or the third step:

pharmaceutical compositions and methods of administration

The invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of the compound.

Because the compound of the present invention has excellent antitumor activity, the compound of the present invention and various crystalline forms thereof, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates thereof, and pharmaceutical compositions containing the compound of the present invention as a main active ingredient are useful for the treatment, prevention and alleviation of diseases associated with tumors.

The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical compositions contain 1-2000mg of the compound of the invention per dose, more preferably 10-1000mg of the compound of the invention per dose. Preferably, the "one dose" is a capsule or tablet.

"pharmaceutically acceptable carrier" means: one or more compatible solid or liquid filler or gel materials which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. "compatible" as used herein means that the components of the composition are capable of blending with and between the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g. sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g. stearic acid, magnesium stearate), calcium sulphate, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g. propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiersWetting agents (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizing agents, antioxidants, preservatives, pyrogen-free water and the like。

The pharmaceutical composition is injection, capsule, tablet, pill, powder or granule.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) Fillers or compatibilizers, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) Binders, for example, hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, e.g., glycerin; (d) Disintegrants, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) a slow solvent, such as paraffin; (f) an absorption accelerator, e.g., a quaternary amine compound; (g) Wetting agents, such as cetyl alcohol and glycerol monostearate; (h) an adsorbent, for example, kaolin; and (i) a lubricant, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be released in a delayed manner in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. The active compound may also be in the form of microcapsules with one or more of the above excipients, if desired.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of these substances and the like.

In addition to these inert diluents, the compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

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

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

Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the invention may be administered alone or in combination with other pharmaceutically acceptable compounds (e.g., antineoplastic agents).

The methods of treatment of the present invention may be administered alone or in combination with other therapeutic means or therapeutic agents.

When a pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is applied to a mammal (e.g., a human) in need of treatment, wherein the dose at the time of administration is a pharmaceutically effective dose, and the daily dose is usually 1 to 2000mg, preferably 50 to 1000mg, for a human having a body weight of 60 kg. Of course, the particular dosage should also take into account factors such as the route of administration, the health of the patient, etc., which are within the skill of the skilled practitioner.

Compared with the prior art, the invention has the following main advantages:

(1) The compound has excellent PAK4 inhibitory activity and excellent water solubility;

(2) The compound has excellent safety, pharmacokinetic performance and PAK4 selective inhibition;

(3) The compound has excellent tumor cell proliferation inhibition activity.

The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental procedure, which does not address the specific conditions in the examples below, is generally followed by routine conditions such as Sambrook et al, molecular cloning: conditions described in the laboratory Manual (New York: cold Spring Harbor Laboratory Press, 1989) or as recommended by the manufacturer. Percentages and parts are by weight unless otherwise indicated.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred methods and materials described herein are presented for illustrative purposes only.

¹ H-NMR was performed using a model Varian MercuryAMX and Varian MR-400; reagents were purchased from J&K Chemica carboline chemical reagent Co., shaoshao far technology (Shanghai) Co., shanghai Bi De medical technology Co., shanghai hong biological medicine technology Co., ltd., and the rest of the reagents are purchased from national pharmaceutical group chemical reagent Co., ltd. Redistilling all solvents before use, wherein the anhydrous solvents are obtained by drying according to a standard method; except for the description, all reactions were performed under nitrogen protection and followed by TLC, after-treatment by washing with saturated aqueous sodium chloride solution and drying with anhydrous sodium sulfate; purification of the product was performed by silica gel (200-300 mesh) column chromatography unless otherwise specified; wherein the silica gel (200-300 mesh) is prepared from Qingdao The GF-254 thin-layer silica gel plate is produced by Katao Jiang silicon gel development Co.

Preparation example 1: preparation of Compound S1

Synthesis of Compounds 1-2:

compound 1-1 (4.62 g,19.6 mmol) was dissolved in tetrahydrofuran (40 mL), cooled to 0deg.C, 1mol/L of a tetrahydrofuran solution of sodium borohydride tetrahydrofuran complex (39.2 mL,39.2 mmol) was carefully added, stirred for 20 minutes, heated to reflux, and stirred for 3 hours. After the TLC detection, the reaction is cooled to room temperature, quenched by adding a proper amount of methanol, and concentrated under reduced pressure. Ethyl acetate was added for dissolution, extraction was performed three times with 2M hydrochloric acid solution, then excess ammonia water was added for basification, extraction was performed three times with dichloromethane, and the organic layer was washed with water, saturated brine, and dried over anhydrous sodium sulfate in this order. Suction filtration and concentration of the filtrate under reduced pressure gave compound 1-2 (2.2 g, 47%). ¹ H NMR (400 MHz, deuterated chloroform) delta 7.68 (d, j=1.7 hz, 1H), 7.47 (s, 1H), 7.36 (dt, j=17.6, 5.2hz, 2H), 3.03 (t, j=6.8 hz, 2H), 2.78 (t, j=6.8 hz, 2H).

Synthesis of Compounds 1-3:

compound 1-2 (2 g,8.33 mmol) was dissolved in formic acid (20 mL), paraformaldehyde (750 mg,8.33 mmol) was added, heated to 50deg.C, and stirred for three hours (continuous monitoring of the reaction). After the TLC detection, the reaction is cooled to room temperature, and 1mol/L sodium hydroxide aqueous solution is added to adjust the pH value to be neutral. Ethyl acetate was added to the mixture to extract, and the organic layer was washed with water and then with saturated brine, followed by drying over anhydrous sodium sulfate. Suction filtration and concentration of the filtrate under reduced pressure to obtain crude product 1-3, which is directly used in the next step without further treatment.

Synthesis of Compounds 1-4:

the crude product 1-3 was dissolved in dichloromethane (20 mL), cooled to 0deg.C, di-tert-butyl dicarbonate (2.73 g,12.5 mmol), N, N-diisopropylethylamine (2.15 g,16.66 mmol) was added and stirred overnight. TLC detection was complete. Extracting with ethyl acetate, and sequentially adding water and saturated saline into the organic layerWashed and dried over anhydrous sodium sulfate. Suction filtration, concentration of the filtrate under reduced pressure, and column chromatography separation and purification gave compound 1-4 (1.46 g, 50%). ¹ H NMR (400 MHz, deuterated chloroform) delta 7.55 (s, 1H), 7.36-7.27 (m, 2H), 4.58 (s, 2H), 3.74 (s, 2H), 2.68 (s, 2H), 1.50 (s, 9H).

Synthesis of Compounds 1-6:

compounds 1-4 (85 mg,0.24 mmol), 1-5 (94 mg,0.266 mmol), potassium carbonate (83 mg,0.6 mmol) were dissolved in a mixed solution of 1, 4-dioxane (2 mL) and water (0.5 mL), and tetrakis (triphenylphosphine) palladium (28 mg,0.024 mmol) was added under nitrogen. Heated to reflux and stirred for 2 hours. After completion of TLC detection, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. Ethyl acetate was added to the mixture to extract, and the organic layer was washed with water and then with saturated brine, followed by drying over anhydrous sodium sulfate. Suction filtration, concentration of the filtrate under reduced pressure, column chromatography separation and purification gave compound 1-6 (105 mg, 88%). ¹ H NMR (400 MHz, deuterated chloroform) delta 7.82 (q, j=8.4 hz, 4H), 7.67 (s, 1H), 7.61-7.47 (m, 2H), 4.65 (s, 2H), 3.88-3.75 (m, 6H), 3.13-2.99 (m, 4H), 2.79 (s, 2H), 1.52 (s, 9H).

Synthesis of Compounds 1-7:

compounds 1 to 6 (105 mg,0.21 mmol) were dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added thereto and stirred at room temperature for 2 hours. After the TLC detection, the reaction was completed, and concentrated under reduced pressure to give crude compound 1-7, which was used directly in the next step without further treatment.

Synthesis of compound S1:

compounds 1 to 7 (84 mg,0.215 mmol), 1 to 8 (39 mg,0.237 mmol) and 2- (7-azobenzotriazole-N, N, N ', N' -tetramethylurea hexafluorophosphate (HATU, 122mg,0.32 mmol) were dissolved in N, N-dimethylformamide (1 mL), N, N-diisopropylethylamine (139 mg,1.08 mmol) was added, and the mixture was stirred at room temperature overnight. ¹ H NMR(500MHz,DMSO-d ₆ )δ8.20(s,1H),8.01(d,J＝8.3Hz,2H),7.97(s,1H),7.92(d,J＝7.9Hz,1H),7.82(d,J＝8.3Hz,2H),7.70(q,J＝8.6Hz,2H),7.46(d,J＝15.2Hz,1H),7.16(d,J＝15.0Hz,1H),6.49(d,J＝8.7Hz,1H),6.44(s,2H),4.92(d,J＝96.5Hz,2H),4.00(d,J＝45.3Hz,2H),3.70–3.60(m,4H),2.98–2.75(d,J＝38.4Hz,6H).

Preparation example 2: preparation of Compound S2

Synthesis of Compound 2-2:

compounds 1-4 (76 mg,0.216 mmol), 2-1 (81 mg,0.238 mmol), potassium carbonate (75 mg,0.54 mmol) were dissolved in a mixed solution of 1, 4-dioxane (2 mL) and water (0.5 mL), and tetrakis (triphenylphosphine) palladium (26 mg,0.022 mmol) was added under nitrogen. Heated to reflux and stirred for 2 hours. After completion of TLC detection, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. Ethyl acetate was added to the mixture to extract, and the organic layer was washed with water and then with saturated brine, followed by drying over anhydrous sodium sulfate. Suction filtration, concentration of the filtrate under reduced pressure, column chromatography separation and purification gave compound 2-2 (75 mg, 71%). ¹ H NMR (400 MHz, deuterated chloroform) delta 7.85 (d, j=8.4 hz, 2H), 7.74-7.61 (m, 3H), 7.55-7.45 (m, 2H), 4.64 (s, 2H), 3.85-3.73 (m, 4H), 2.77 (s, 2H), 2.59-2.44 (m, 2H), 1.53 (s, 9H).

Synthesis of Compound 2-3:

compound 2-2 (75 mg,0.15 mmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the TLC detection, the reaction was completed, and concentrated under reduced pressure to give crude compound 2-3, which was used directly in the next step without further treatment.

Synthesis of compound S2:

compound 2-3 (60 mg,0.15 mmol), 1-8 (27 mg,0.165 mmol), HATU (94 mg,0.25 mmol) were dissolved in N, N-dimethylformamide (1 mL), N-diisopropylethylamine (97 mg,0.75 mmol) was added, and stirred at room temperature overnight. After completion of TLC detection, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. Ethyl acetate was added to the mixture to extract, and the organic layer was washed with water and then with saturated brine, followed by drying over anhydrous sodium sulfate. Suction filtration, concentration of the filtrate under reduced pressure, column chromatography separation and purification gave compound S2 (28 mg, 35%) as a white solid. ¹ H NMR(600MHz,DMSO-d ₆ )δ8.74(t,J＝5.6Hz,1H),8.19(s,1H),7.97–7.88(m,4H),7.83(d,J＝8.3Hz,2H),7.65(q,J＝8.6Hz,2H),7.45(d,J＝15.1Hz,1H),7.20–7.05(m,1H),6.58–6.36(m,3H),4.90(d,J＝120.1Hz,2H),3.99(d,J＝60.2Hz,2H),3.53(dd,J＝12.7,6.7Hz,2H),2.80(d,J＝42.1Hz,2H),2.66–2.52(m,2H).

Preparation example 3: preparation of Compound S3

Synthesis of Compound 3-2:

compound 3-1 (3 g,16.1 mmol), N-bromosuccinimide (3.45 g,19.4 mmol) was dissolved in acetonitrile (35 mL) and stirred at room temperature for 4 hours. After completion of TLC detection, the reaction was cooled to 0℃and ice water was added thereto to precipitate a solid, which was filtered and dried to give Compound 3-2 (3.98 g, 93%). ¹ H NMR (400 MHz, deuterated chloroform) δ11.28 (s, 1H), 7.93-7.87 (m, 1H), 7.68 (d, j=2.4 hz, 1H), 3.98 (s, 3H).

Synthesis of Compound 3-3:

compound 3-2 (1.1 g,4.14 mmol), ethyl bromoacetate (830 mg,4.97 mmol), potassium carbonate (1.71 g,12.42 mmol) were dissolved in acetone (20 mL), heated to reflux, and stirred for 4 hours. After completion of TLC detection, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. Ethyl acetate was added to the mixture to extract, and the organic layer was washed with water and then with saturated brine, followed by drying over anhydrous sodium sulfate. Suction filtration and concentration of the filtrate under reduced pressure gave compound 3-3 (1.45 g, 99%). ¹ H NMR (400 MHz, deuterated chloroform) δ7.83 (d, j=2.5 hz, 1H), 7.69 (d, j=2.5 hz, 1H), 4.66 (s, 2H), 4.30 (q, j=7.1 hz, 2H), 3.90 (s, 3H), 1.32 (t, j=7.1 hz, 4H).

Synthesis of Compounds 3-4:

compound 3-3 (1.45 g,4.1 mmol) was dissolved in a mixture of 1M aqueous sodium hydroxide (50 mL) and ethanol (50 mL), heated to reflux, and reacted for 6 hours. After the TLC detection, the reaction was completed, cooled to 0℃and then added with hydrochloric acid solution to adjust the pH to acidity, and then a solid was precipitated and suction-filtered to obtain Compound 3-4 (1.09 g, 86%). ¹ H NMR (400 MHz, deuterated methanol) δ7.81 (d, j=2.3 hz, 1H), 7.77 (d, j=2.4 hz, 1H), 4.73 (s, 2H).

Synthesis of Compounds 3-5:

compound 3-4 (1.09 g,3.52 mmol) was dissolved in a mixed solution of acetic acid (5 mL) and acetic anhydride (15 mL), heated to 130℃and reacted overnight. After completion of the TLC detection, the reaction was cooled to room temperature, a proper amount of water was added, extraction was performed 3 times with ethyl acetate and petroleum ether (1:1), the organic layers were combined, washed 5 times with aqueous sodium bicarbonate, then with saturated brine, and dried over anhydrous sodium sulfate. Suction filtration, and decompression concentration of filtrate to obtain coarse product 3-5.

Synthesis of Compounds 3-6:

the crude product 3-5 was dissolved in a mixed solution of methanol (5 mL) and 1M aqueous hydrochloric acid (2 mL), heated to reflux, and reacted for 3 hours. After the TLC detection, the reaction was cooled to room temperature, and an appropriate amount of ice water was added thereto to precipitate a solid, which was suction-filtered to give Compound 3-6 (697 mg, 80%). ¹ H NMR (400 MHz, deuterated chloroform) delta 7.75 (d, j=1.8 hz, 1H), 7.71 (d, j=1.8 hz, 1H), 4.76 (s, 2H).

Synthesis of Compounds 3-7:

compound 1-2 (1.77 g,10 mmol) was dissolved in tetrahydrofuran (20 mL), cooled to 0deg.C, sodium hydride (480 mg,12 mmol) was added carefully and stirred for one hour. Tetrahydrofuran (20 mL) dissolved in 3-6 (10 mmol,2.47 g) was carefully added and stirred overnight. After completion of TLC detection, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. Ethyl acetate was added to the mixture to extract, and the organic layer was washed with water and then with saturated brine, followed by drying over anhydrous sodium sulfate. Suction filtration, concentration of filtrate under reduced pressure, column chromatography separation and purification to obtain compound 3-7 (2.33 g, 86%). ¹ H NMR (400 MHz, deuterated chloroform) delta 7.75 (s, 1H), 7.64 (d, j=1.7 hz, 1H), 7.52 (d, j=1.5 hz, 1H), 3.74 (d, j=1.2 hz, 2H).

Synthesis of Compounds 3-8:

synthesis of Compounds 3-8 reference Synthesis of Compounds 1-4. ¹ H NMR (600 MHz, deuterated chloroform) delta 7.60 (d, j=1.6 hz, 1H), 7.55 (s, 1H), 7.44 (d, j=1.6 hz, 1H), 3.03 (t, j=6.8 hz, 2H), 2.78 (t, j=6.7 hz, 2H).

Synthesis of Compounds 3-10:

synthesis of Compounds 3-10 reference Synthesis of Compounds 1-6. ¹ H NMR (400 MHz, deuterated chloroform) delta 7.47 (s, 1H),7.39(s,1H),4.63(s,2H),3.75(s,2H),2.68(s,2H),1.50(s,9H).

synthesis of Compounds 3-12:

synthesis of Compounds 3-12 reference Synthesis of Compounds 1-6. ¹ H NMR (400 MHz, deuterated chloroform) delta 7.64 (d, j=8.1 hz, 2H), 7.55-7.46 (m, 4H), 4.66 (s, 2H), 4.01-3.53 (m, 6H), 2.76 (s, 2H), 2.16-1.87 (m, 4H), 1.50 (s, 9H).

Synthesis of Compounds 3-13: synthesis of Compounds 3-13 reference Synthesis of Compounds 1-7.

Synthesis of compound S3:

synthesis of compound S3 reference synthesis of compound S1. White solid. ¹ H NMR(600MHz,DMSO-d ₆ )δ8.18(s,1H),7.91(d,J＝8.3Hz,1H),7.88(s,1H),7.81(d,J＝8.1Hz,2H),7.72(s,1H),7.54(d,J＝8.1Hz,2H),7.44(d,J＝15.1Hz,1H),7.18–7.06(m,1H),6.48(s,3H),5.05(s,1H),4.83(s,1H),3.97(d,J＝63.0Hz,2H),3.59(d,J＝156.5Hz,4H),3.32(s,1H),2.79(d,J＝40.9Hz,2H),2.13–1.96(m,4H).

Preparation example 4: preparation of Compound S4

Synthesis of compound S4: the same as in preparation example 3, except that: 3-11 is replaced by

A white solid was obtained. ¹ H NMR (500 MHz, deuterated methanol) δ8.11 (s, 1H), 7.90 (dd, j=8.8, 2.2hz, 1H), 7.76-7.70 (m, 4H), 7.68 (s, 1H), 7.55 (d, j=14.7 hz, 2H), 7.12-6.97 (m, 1H), 6.63 (d, j=8.8 hz, 1H), 5.03-4.82 (m, 5H), 4.16-4.00 (m, 4H), 3.89 (s, 2H), 2.85 (d, j=20.9 hz, 2H), 1.34 (s, 6H).

Preparation example 5: preparation of Compound S5

Synthesis of compound S5: the same as in preparation example 3, except that: 3-11 is replaced by

A white solid was obtained. ¹ H NMR (500 MHz, deuterated methanol) delta 8.10 (s, 1H), 7.90 (dd, j=8.8, 2.3hz, 1H), 7.71 (t, j=7.1 hz, 2H), 7.68-7.59 (m, 3H), 7.55 (dd, j=8.3, 7.0hz, 2H), 7.13-6.98 (m, 1H), 6.67-6.60 (m, 1H), 4.99-4.86 (m, 2H), 4.04 (t, j=5.3 hz, 2H), 3.94-3.58 (m, 5H), 2.84 (d, j=19.5 hz, 2H), 2.34-1.98 (m, 3H), 1.67-1.48 (m, 3H).

Preparation example 6: preparation of compound S6:

synthesis of compound S6: the same as in preparation example 3, except that: 3-11 is replaced by

A white solid was obtained. ¹ H NMR (500 MHz, deuterated methanol) δ8.10 (d, j=1.7hz, 1H), 7.90 (dd, j=8.8, 2.3hz, 1H), 7.64-7.59 (m, 2H), 7.58-7.53 (m, 1H), 7.49 (s, 1H), 7.38-7.34 (m, 2H), 7.11-6.99 (m, 1H), 6.63 (d, j=8.8hz, 1H), 5.05-4.87 (m, 2H), 4.06 (s, 2H), 3.95-3.53 (m, 4H), 2.92-2.77 (m, 2H), 2.19-1.96 (m, 4H).

Preparation example 7: preparation of Compound S7

Synthesis of Compound 7-3

To a mixed solution of compound 7-1 (9.28 g,40 mmol) and 7-2 (6.16 g,44 mmol) in N, N-dimethylformamide (40 mL) and water (20 mL), potassium phosphate (25.44 g,120 mmol) and [1,1' -bis (diphenylphosphino) ferrocene were added]Palladium dichloride (1.46 g,2 mmol), nitrogen-blanketed, was heated to 45℃and stirred for 5 hours. After completion of TLC detection, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. Ethyl acetate was added to the mixture to extract, and the organic layer was washed with water and then with saturated brine, followed by drying over anhydrous sodium sulfate. Suction filtration, concentration of the filtrate under reduced pressure, and column chromatography separation and purification gave compound 7-3 (5.3 g, 54%). ¹ H NMR (400 MHz, deuterated chloroform) δ11.30 (s, 1H), 7.85 (d, j=7.9 hz, 1H), 7.54 (dd, j=8.3, 5.7hz, 2H), 7.48 (d, j=7.2 hz, 1H), 7.11 (t, j=8.6 hz, 2H), 6.94 (t, j=7.8 hz, 1H), 3.96 (s, 3H).

Synthesis of Compound 7-4

Compound 7-3 (3.1 g,12.6 mmol) and N-bromosuccinimide (2.7 g,15.1 mmol) were dissolved in acetonitrile (30 mL), heated to 60℃and stirred at room temperature for 4 hours. After completion of TLC detection, the reaction was cooled, ice-water was added thereto, and a solid was precipitated, filtered and dried to give Compound 7-4 (3.8 g, 93%). ¹ H NMR (400 MHz, deuterated chloroform) δ11.24 (s, 1H), 7.96 (d, j=2.5 hz, 1H), 7.58 (d, j=2.1 hz, 1H), 7.51 (dd, j=8.9, 5.4hz, 2H), 7.11 (t, j=8.8 hz, 2H), 3.97 (s, 3H).

Synthesis of Compound S7

The subsequent steps are described with reference to preparation example 3.

A white solid was obtained. ¹ H NMR (500 MHz, deuterated methanol) δ8.10 (s, 1H), 7.90 (s, 3H), 7.79 (d, j=8.2 hz, 2H), 7.71 (s, 1H), 7.65 (s, 1H), 7.60-7.50 (m, 3H), 7.24 (t, j=8.7 hz, 2H), 7.05 (dd, j=37.5, 15.3hz, 1H), 6.65 (d, j=8.0 hz, 1H), 5.00-4.86 (m, 2H), 4.14-4.02 (m, 2H), 3.76 (d, j=112.2 hz, 4H), 2.87 (d, j=21.4 hz, 2H), 2.08 (s, 4H).

Preparation example 8: preparation of Compound S8

Synthesis of compound S8: the same as in preparation example 7 is distinguished in that: 3-11 is replaced by

A white solid was obtained. ¹ H NMR (500 MHz, deuterated methanol) delta 8.02 (d, j=28.0 hz, 1H), 7.89-7.74 (m, 3H), 7.68 (d, j=8.0 hz, 2H), 7.63-7.43 (m, 5H), 7.17 (q, j=8.6 hz, 2H), 6.96 (dd, j=40.8, 15.2hz, 1H), 6.65-6.52 (m, 1H), 4.84-4.71 (m, 2H), 3.96 (s, 2H), 3.90-3.53 (m, 4H), 2.77 (s, 2H), 2.29-1.95 (m, 2H), 1.53 (dd, j=46.5, 20.6hz, 3H).

Preparation example 9: preparation of Compound S9

Synthesis of compound S9: the same as in preparation example 7 is distinguished in that: 3-11 is replaced by 1-5.

A white solid was obtained. ¹ H NMR (500 MHz, deuterated chloroform) delta 8.23 (s, 1H), 7.89-7.79 (m, 6H), 7.72-7.55 (m, 4H), 7.22 (t, j=8.7 hz, 2H), 6.81 (s, 1H), 6.53 (d, j=8.6Hz,1H),4.93(s,2H),4.04(d,J＝44.4Hz,2H),3.78–3.74(m,4H),3.10–3.02(m,4H),2.91(s,2H).

Preparation example 10: preparation of Compound S10

Synthesis of compound S10: the same as in preparation example 7 is distinguished in that: 3-11 is replaced by

A white solid was obtained. ¹ H NMR (600 MHz, deuterated chloroform) delta 8.23 (s, 1H), 7.83 (s, 2H), 7.73-7.62 (m, 2H), 7.58 (t, j=7.7 hz, 2H), 7.53 (s, 1H), 7.31-7.27 (m, 2H), 7.21 (t, j=8.6 hz, 2H), 4.90 (d, j=38.0 hz, 4H), 4.19-3.53 (m, 5H), 2.93 (d, j=33.0 hz, 2H), 2.08-1.83 (m, 3H).

Preparation example 11: preparation of Compound S11

Synthesis of Compound S11

Compound 11-1 (223 mg,1 mmol) and 11-2 (221 mg,1.1 mmol) were dissolved in dichloromethane (5 mL), pyridine (158 mg,2 mmol) was added, and the mixture was stirred at room temperature for 5 hours. After completion of TLC detection, ethyl acetate was added for extraction, and the organic layer was washed with water, saturated brine, and dried over anhydrous sodium sulfate. Suction filtration, concentration of filtrate under reduced pressure, column chromatography separation and purification to obtain compound 11-3 (330 mg, 85%). ¹ H NMR (400 MHz, deuterated chloroform) delta 8.28-8.21 (m, 3H), 7.96 (d, j=8.9 hz, 1H), 7.67 (d, j=6.6 hz, 1H), 7.33 (d, j=9.0 hz, 2H), 5.44 (s, 1H), 4.41 (d, j=6.0 hz, 2H), 1.53 (s, 9H).

Compound 11-3 (52 mg,0.133 mmol) and 3-13a (52 mg,0.12 mmol) were dissolved in tetrahydrofuran (2 mL), N-diisopropylethylamine (46 mg,0.36 mmol) was added, and the mixture was stirred at room temperature overnight. After completion of TLC detection, ethyl acetate was added for extraction, and the organic layer was washed with water, saturated brine, and dried over anhydrous sodium sulfate. Suction filtration, concentration of the filtrate under reduced pressure, and column chromatography separation and purification (methanol: dichloromethane=2.5%) gave compound 11-4 (50 mg, 61%). ¹ H NMR (400 MHz, deuterated chloroform) delta 8.23 (s, 1H), 8.18 (s, 1H), 7.90 (d, j=8.5 hz, 1H), 7.66 (d, j=8.6 hz, 1H), 7.59 (d, j=8.1 hz, 2H), 7.49-7.42 (m, 4H), 4.59 (s, 2H), 4.40 (d, j=5.4 hz, 2H), 4-3.51 (m, 6H), 2.73 (s, 2H), 2.14-1.90 (m, 4H), 1.52 (s, 9H).

Compound 11-4 (50 mg,0.073 mmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL), and stirred at room temperature overnight. After completion of TLC detection, ethyl acetate was added for extraction, and the organic layer was washed with water, saturated brine, and dried over anhydrous sodium sulfate. Suction filtration, concentration of the filtrate under reduced pressure, and column chromatography separation and purification (methanol: dichloromethane=5%) gave compound S11 (38 mg, 90%). ¹ H NMR(500MHz,DMSO-d ₆ )δ7.88(d,J＝1.4Hz,1H),7.85–7.81(m,3H),7.73(d,J＝1.5Hz,1H),7.59–7.54(m,2H),7.31(dd,J＝8.4,2.4Hz,1H),7.20(t,J＝5.6Hz,1H),6.39(d,J＝8.4Hz,1H),5.74(s,2H),4.64(s,2H),4.09(d,J＝5.4Hz,2H),3.84–3.38(m,6H),2.74(s,2H),2.07(s,4H).

Preparation example 12: preparation of Compound S12

Synthesis of compound S12: the same as in preparation example 11 was distinguished in that: 3-13a is replaced with 7-14.

A white solid was obtained. ¹ H NMR (500 MHz, deuterated methanol) delta 7.87-7.81 (m, 3H), 7.74 (d, j=8.3 hz, 2H), 7.64 (d, j=1.7 hz, 1H), 7.60-7.57 (m, 2H), 7.53 (d, j=8.3 hz, 2H), 7.23-7.17 (m, 2H), 6.65 (d, j=8.7 hz, 1H), 4.59 (s, 2H), 4.28 (d, j=23.6 hz, 2H), 3.98-3.50 (m, 6H), 2.76 (s, 2H), 2.07 (bs, 4H).

Preparation example 13: preparation of Compound S13

Synthesis of compound S13: the same as in preparation example 11 was distinguished in that: 11-3 is replaced with 13-2.

A white solid was obtained. ¹ H NMR (500 MHz, deuteriumMethyl alcohol), delta 8.51 (d, j=19.3 hz, 1H), 8.41 (s, 1H), 7.89-7.80 (m, 3H), 7.77-7.71 (m, 2H), 7.63 (d, j=1.8 hz, 1H), 7.57 (d, j=1.8 hz, 1H), 7.54-7.50 (m, 2H), 4.61 (s, 2H), 4.44 (s, 2H), 4.00-3.51 (m, 7H), 2.76 (t, j=5.3 hz, 2H), 2.07 (s, 4H).

Preparation example 14: preparation of Compound S14

Compound 7-12 (5.9 g,13.3 mmol) was dissolved in 100mL 1, 4-dioxane and potassium acetate (3.9 g,39.8 mmol), pd (dppf) Cl was added ₂ (4815 mg,0.66 mmol), pinacol diboronate (4 g,15.9 mmol) were heated to reflux and stirred for 3 hours, after completion of TLC detection the reaction was cooled to room temperature, the reaction mixture was diluted with water, extracted three times with ethyl acetate, the organic layer was washed successively with water, saturated brine and dried over anhydrous sodium sulfate. Suction filtration and concentration of the filtrate under reduced pressure gave compound 14-1 (3.5 g, 47%). ¹ H NMR(400MHz,Chloroform-d)δ7.90(s,1H),7.86–7.78(m,3H),7.21–7.13(m,2H),4.61(s,2H),3.77(s,2H),2.76(s,2H),1.50(s,9H),1.38(s,12H).

A mixed solution of compound 14-1 (50 mg,0.1 mmol) in 1, 4-dioxane (0.9 mL) and water (0.3 mL) was added Pd (PPh) ₃ ) ₄ (12mg,0.01mmol)，K ₂ CO ₃ (35 mg,0.25 mmol) was refluxed at 100℃for 1 hour. After the TLC detection, the reaction solution was cooled to room temperature, diluted with water, extracted three times with ethyl acetate, and the organic layer was washed with water, saturated brine, and dried over anhydrous sodium sulfate. Suction filtration and concentration of the filtrate under reduced pressure gave compound 14-3 (50 mg, 83%). ¹ H NMR (400 MHz, deuterated chloroform) delta 8.88 (s, 1H), 8.07 (dd, j=8.1, 2.1hz, 1H), 7.88-7.77 (m, 3H), 7.61 (s, 1H), 7.55 (s, 1H), 7.22 (t, j=8.6 hz, 2H), 4.65 (s, 2H), 3.94 (t, j=5.8 hz, 2H), 3.81 (s, 2H), 2.81 (s, 2H), 2.22-2.08 (m, 4H), 1.51 (s, 9H).

Compound 14-3 (50 mg,0.08 mmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 30 minutes, and the solvent was removed to give compound 14-4. 14-4 was dissolved in DMF (1 mL) and Compounds 1-8 (16 mg,0.1 mmol) were added, HATU (47 mg,0.12 mm)ol), DIPEA (112 μl,0.64 mmol). The reaction was carried out at room temperature overnight, the reaction mixture was diluted with water, extracted three times with ethyl acetate, and the organic layer was washed with water, saturated brine and dried over anhydrous sodium sulfate. Suction filtration and concentration of the filtrate under reduced pressure gave compound S14 (20 mg, 37%). ¹ H NMR (400 MHz, deuterated chloroform) delta 8.88 (d, j=1.8 hz, 1H), 8.24 (s, 1H), 8.08 (dd, j=8.1, 2.3hz, 1H), 7.88-7.78 (m, 3H), 7.71-7.60 (m, 3H), 7.57 (d, j=1.8 hz, 1H), 7.22 (t, j=8.6 hz, 2H), 6.88-6.70 (m, 1H), 6.53 (d, j=8.6 hz, 1H), 4.99-4.73 (m, 4H), 4.15-3.97 (m, 2H), 3.94 (t, j=5.9 hz, 2H), 3.81 (t, j=5.9 hz, 2H), 2.91 (s, 2H), 2.22-2.03 (m, 4H).

Preparation example 15: preparation of Compound S15

Synthesis of compound S15: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (500 MHz, deuterated methanol) δ8.74 (d, j=2.0 hz, 1H), 8.09 (s, 3H), 8.03 (d, j=8.2 hz, 1H), 7.98-7.83 (m, 4H), 7.55 (d, j=15.3 hz, 1H), 7.24 (t, j=8.4 hz, 2H), 7.04 (dd, j=33.8, 14.7hz, 1H), 6.63 (d, j=7.9 hz, 1H), 5.00-4.85 (m, 2H), 4.09-4.00 (m, 2H), 3.79 (d, j=115.9 hz, 4H), 2.92-2.78 (m, 2H), 2.23-1.96 (m, 4H).

Preparation example S16: preparation of Compound S16

Synthesis of compound S16: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated chloroform) δ9.07-9.01 (m, 2H), 8.22 (s, 1H), 8.15-8.03 (m, 2H), 7.85 (t, j=6.5 hz, 2H), 7.72-7.55 (m, 2H), 7.21 (t, j=8.6 hz, 2H), 6.87-6.68 (m, 1H), 6.51 (d, j=8.7 hz, 1H), 5.02-4.62 (m, 4H), 4.15-3.91 (m, 4H), 3.82 (s, 2H), 2.91 (s, 2H), 2.21-2.04 (m, 4H).

Preparation example S17: preparation of Compound S17

Synthesis of compound S17: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated chloroform) δ9.08 (s, 2H), 8.23 (s, 1H), 7.83 (dd, j=8.5, 5.3hz, 2H), 7.71-7.59 (m, 3H), 7.54 (d, j=1.8 hz, 1H), 7.23 (t, j=9.1 hz, 2H), 6.89-6.69 (m, 1H), 6.52 (d, j=8.6 hz, 1H), 5.00-4.86 (m, 2H), 4.82 (s, 2H), 4.15-3.93 (m, 4H), 3.56 (t, j=5.9 hz, 2H), 2.92 (s, 2H), 2.22-2.02 (m, 4H).

Preparation example S18: preparation of Compound S18

Synthesis of compound S18: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated chloroform) delta 8.89 (s, 2H), 8.62-8.54 (m, 2H), 8.24 (s, 1H), 7.94-7.86 (m, 2H), 7.74-7.58 (m, 2H), 7.22 (t, j=8.5 hz, 2H), 6.88-6.69 (m, 1H), 6.52 (d, j=8.6 hz, 1H), 5.06-4.53 (m, 4H), 4.22-3.58 (m, 6H), 2.93 (s, 2H), 2.09 (s, 4H).

Preparation example S19: preparation of Compound S19

Synthesis of compound S19: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (700 MHz, deuterated chloroform) delta 8.25 (d, j=19.7 hz, 2H), 8.18-8.06 (m, 2H), 8.01 (d, j=9.0 hz, 1H), 7.92-7.83 (m, 2H), 7.74-7.59 (m, 2H), 7.22 (t, j=8.3 hz, 2H), 6.87-6.69 (m, 1H), 6.52 (d, j=8.5 hz, 1H), 5.00-4.86 (m, 2H), 4.81 (s, 2H), 4.16-3.94 (m, 6H), 2.92 (s, 2H), 2.31-2.10 (m, 4H).

Preparation example S20: preparation of Compound S20

Synthesis of compound S20: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated chloroform) delta 8.25 (s, 2H), 8.17-8.05 (m, 3H), 7.92-7.84 (m, 2H), 7.66 (d, j=14.8 hz, 2H), 7.23 (t, j=9.6 hz, 2H), 6.88-6.70 (m, 1H), 6.52 (d, j=8.6 hz, 1H), 4.93 (d, j=11.3 hz, 2H), 4.70 (s, 2H), 4.17 (dt, j=12.4, 6.3hz, 4H), 4.12-3.93 (m, 2H), 2.93 (s, 2H), 2.74 (t, j=6.2 hz, 2H), 2.69 (t, j=6.4 hz, 2H).

Preparation example S21: preparation of Compound S21

Synthesis of compound S21: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated chloroform) δ8.22 (d, j=6.7hz, 2H), 8.10 (d, j=8.8hz, 2H), 7.98 (d, j=8.8hz, 1H), 7.90-7.83 (m, 2H), 7.71-7.60 (m, 2H), 7.21 (t, j=8.6hz, 2H), 6.87-6.67 (m, 1H), 6.51 (d, j=8.6hz, 1H), 5.01-4.75 (m, 5H), 4.45 (d, j=14.0hz, 1H), 4.03 (d, j=33.8hz, 2H), 3.52-3.43 (m, 1H), 3.20 (td, j=13.2, 2.7hz, 1H), 2.91 (s, 2H), 2.26-1.96 (m, 5H), 5.01-4.75 (m, 5H), 4.45 (d, j=14.0hz, 1H), 3.52-3.43 (m, 1H), 3.20 (td, j=13.2.7hz, 1H).

Preparation example S22: preparation of compound S22:

synthesis of compound S22: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated chloroform) delta 8.31-8.03%m,5H),7.91–7.83(m,2H),7.73–7.60(m,2H),7.21(t,J＝8.5Hz,2H),6.90–6.66(m,1H),6.52(d,J＝8.7Hz,1H),5.04–4.62(m,4H),4.56(d,J＝12.3Hz,1H),4.40(dd,J＝33.7,12.8Hz,2H),4.19–3.95(m,3H),2.92(s,2H),2.53–2.29(m,2H).

Preparation example S23: preparation of Compound S23

Synthesis of compound S23: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (700 MHz, deuterated dimethyl sulfoxide) delta 8.98 (d, j=9.3 hz, 1H), 8.63 (d, j=8.8 hz, 1H), 8.43 (s, 1H), 8.33 (d, j=1.8 hz, 1H), 8.28 (d, j=8.8 hz, 1H), 8.19 (d, j=7.3 hz, 1H), 8.04-7.99 (m, 2H), 7.92 (d, j=8.6 hz, 1H), 7.49-7.44 (m, 1H), 7.42 (t, j=8.7 hz, 2H), 7.14 (dd, j=41.1, 15.3hz, 1H), 6.52-6.41 (m, 3H), 5.05 (s, 1H), 4.85 (s, 1H), 4.50-4.37 (m, 1H), 4.02 (d, j=8.6 hz, 1H), 7.49-7.44 (m, 1H), 7.42 (t, j=8.7.42 (m, 2H), 7.14 (d, j=41.3 hz, 15.3H), 6.52-6.41 (m, 1H), 4.52-4.37 (d, 3H), 4.05 (d, 3H), 9.9-2H), 2.9 (2H), 2.9-7.9 (2H), 2.9 (d, 1H), 2.9.7.7 (2H).

Preparation example S24: preparation of Compound S24

Synthesis of compound S24: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated chloroform) δ8.31 (d, j=8.9 hz, 1H), 8.24 (d, j=14.8 hz, 2H), 8.11 (d, j=8.8 hz, 2H), 7.86 (dd, j=8.4, 5.4hz, 2H), 7.72-7.58 (m, 2H), 7.22 (t, j=8.5 hz, 2H), 6.88-6.67 (m, 1H), 6.52 (d, j=8.6 hz, 1H), 6.06 (d, j=4.2 hz, 1H), 5.03 (t, j=9.9 hz, 1H), 4.97-4.82 (m, 5H), 4.44-4.36 (m, 1H), 4.30 (dd, j=11.0, 5.4, 1H), 4.04 (d, j=34.8 hz, 3.28-1H), 6.06 (d, 1H), 3.91 (m, 2H).

Preparation example S25: preparation of Compound S25

Synthesis of compound S25: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (700 MHz, deuterated dimethyl sulfoxide) delta 8.60 (d, j=8.9 hz, 1H), 8.46 (s, 1H), 8.31 (s, 1H), 8.22-8.15 (m, 2H), 8.03 (dd, j=8.4, 5.4hz, 2H), 7.93 (d, j=8.4 hz, 1H), 7.49-7.39 (m, 3H), 7.14 (dd, j=44.7, 15.3hz, 1H), 6.53-6.42 (m, 3H), 5.18 (t, j=5.4 hz, 1H), 4.94 (d, j=139.8 hz, 2H), 4.52-4.43 (m, 2H), 4.07 (s, 1H), 3.97 (s, 2H), 3.96 (s, 1H), 3.90 (d, j=10.1 hz, 1H), 6.53-6.42 (m, 3H), 5.18 (t, j=5.4 hz, 1H).

Preparation example S26: preparation of Compound S26

Synthesis of compound S26: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated dimethyl sulfoxide) delta 8.60 (d, j=8.9 hz, 1H), 8.46 (s, 1H), 8.30 (s, 1H), 8.19 (s, 1H), 8.07-7.99 (m, 3H), 7.93 (d, j=8.8 hz, 1H), 7.50-7.37 (m, 3H), 7.14 (dd, j=24.5, 14.9hz, 1H), 6.51-6.41 (m, 3H), 4.96 (d, j=79.5 hz, 2H), 4.03 (d, j=41.3 hz, 2H), 3.66-3.60 (m, 2H), 3.59-3.53 (m, 2H), 2.87 (d, j=27.0 hz, 2H).

Preparation example S27: preparation of Compound S27

Synthesis of compound S27: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (700 MHz, deuterated dimethyl sulfoxide) delta 8.86 (d, j=8.8 Hz, 1H), 8.70 (s, 1H), 8.55 (s, 1H), 8.44 (s, 1H), 8.27 (dt, j=11.4, 7.1Hz, 3H), 8.18 (d, j=8.7 Hz),1H),7.75–7.63(m,3H),7.40(dd,J＝43.8,14.9Hz,1H),6.74(s,3H),5.20(d,J＝138.4Hz,2H),4.38–3.87(m,6H),3.31–3.01(m,6H),2.84(s,3H).

Preparation example S28: preparation of Compound S28

Synthesis of compound S28: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated dimethyl sulfoxide) delta 9.22-9.09 (m, 1H), 8.63 (d, j=8.9 hz, 1H), 8.44 (s, 1H), 8.32 (d, j=1.8 hz, 1H), 8.26 (d, j=8.8 hz, 1H), 8.19 (s, 1H), 8.02 (dd, j=8.5, 5.5hz, 2H), 7.93 (d, j=8.8 hz, 1H), 7.50-7.36 (m, 3H), 7.23-7.06 (m, 1H), 6.48 (s, 3H), 4.96 (d, j=80.7 hz, 2H), 4.02 (d, j=41.3 hz, 2H), 3.63-3.50 (m, 5H), 3.34 (s, 2H), 2.86 (d, j=27.7 hz, 2H).

Preparation example S29: preparation of Compound S29

Synthesis of compound S29: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated dimethyl sulfoxide) δ9.12 (s, 1H), 8.60 (d, j=8.9 hz, 1H), 8.42 (d, j=1.8 hz, 1H), 8.29 (d, j=1.8 hz, 1H), 8.24 (d, j=8.9 hz, 1H), 8.15 (s, 1H), 8.03-7.96 (m, 2H), 7.89 (d, j=8.8 hz, 1H), 7.49-7.31 (m, 3H), 7.20-7.01 (m, 1H), 6.49-6.34 (m, 3H), 5.02 (s, 1H), 4.82 (s, 1H), 3.99 (d, j=39.9 hz, 2H), 3.63-3.48 (m, 6H), 3.45-3.39 (m, 2H), 3.21 (s, 3.84), 7.20-7.01 (m, 1H), 5.63-3.48 (m, 2H).

Preparation example S30: preparation of Compound S30

Synthesis of compound S30: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated dimethyl sulfoxide) δ11.41 (s, 1H), 8.44 (s, 2H), 8.32 (d, j=1.8 hz, 1H), 8.18 (d, j=2.1 hz, 2H), 8.01 (td, j=5.7, 2.0hz, 2H), 7.93 (d, j=8.8 hz, 1H), 7.50-7.32 (m, 3H), 7.22-7.05 (m, 1H), 6.54-6.38 (m, 3H), 4.94 (d, j=80.4 hz, 2H), 4.02 (d, j=41.3 hz, 2H), 2.97-2.74 (m, 6H).

Preparation example S31: preparation of Compound S31

Synthesis of compound S31: the same as in preparation example 14, except that: substitution of 14-2 for

A white solid was obtained. ¹ H NMR (400 MHz, deuterated dimethyl sulfoxide) delta 10.63 (s, 1H), 8.42-8.31 (m, 2H), 8.25 (s, 1H), 8.18-8.09 (m, 2H), 8.00-7.86 (m, 3H), 7.47-7.32 (m, 3H), 7.17-7.03 (m, 1H), 6.49-6.38 (m, 3H), 4.90 (d, j=79.9 hz, 2H), 4.55 (dd, j=8.2, 5.5hz, 1H), 3.97 (td, j=15.7, 14.9,8.6hz, 3H), 3.82 (q, j=7.1 hz, 1H), 2.81 (d, j=28.0 hz, 2H), 2.28-2.14 (m, 1H), 2.06-1.77 (m, 3H).

Experimental example 1: evaluation of pancreatic cancer cell proliferation inhibitory Activity of Compounds

Sulforhodamine B (SRB) protein staining was used to evaluate the cell proliferation inhibitory activity of the compounds.

1. Pancreatic cancer cells MIAPaCa-2 in the logarithmic growth phase were seeded at an appropriate density in 96-well plates.

Incubated overnight at 2.37℃until cells attached, 10. Mu.L/well of compound at different concentrations was added, 3 secondary wells were set per concentration, and solvent control and cell-free blank control were set.

3. The cells were cultured for 72 hours, the cell culture solution was discarded, 100. Mu.L of a pre-chilled 10% trichloroacetic acid (TCA) solution was added to each well, and the mixture was fixed at 4℃for 1 hour. The fixed liquid is discarded, washed by distilled water and dried in an oven.

4. mu.L of SRB dye was added to each well, and after staining at room temperature for 20min, the dye was discarded. Washing with 1% glacial acetic acid solution, and oven drying.

5. mu.L of 10 mM Tris solution was added to each well to dissolve the SRB dye.

6. The OD value per well was measured with a microplate reader at 560nm wavelength and the compound inhibition ratio was calculated according to the following formula:

inhibition% = (control OD value-dosing OD value)/control OD value x 100%.

TABLE 1 inhibitory Activity of Compounds S19-S23, S25, S26 against MIAPaCa-2 cell proliferation

Cell proliferation inhibition activity test shows that compounds S19-S23, S25 and S26 have better in vitro proliferation inhibition activity on MIAPaCa-2 cells than KPT-9274, wherein, the IC of the compounds S21 and S23 ₅₀ The values were raised by about 4-fold and 3-fold, respectively, over KPT-9274. These results show that the compounds have good tumor cell proliferation inhibition activity, and the in vitro tumor cell proliferation inhibition activity is superior to KPT-9274.

Experimental example 2: determination of Compounds' influence on PAK4 and downstream Signal Activity by immunoblotting experiments

1. U2-OS cells in the logarithmic growth phase were seeded in 6-well plates at an appropriate density, after the cells had attached to the walls, the original culture broth was discarded, 2mL of fresh culture broth was added, and DMSO or each concentration of compound was added, and the cells were cultured at 37℃for 72 hours. The cell culture broth was discarded, washed 3 times with PBS solution, and an appropriate amount of 1X SDS (Sodium Dodecyl Sulfate) cell lysate was added, and after complete cell lysis, the sample was collected to 600. Mu.L EP tube and heated at 100℃for 20min.

2. 10% SDS-PAGE gel (sodium dodecyl sulfate-polyacrylamide gel, sodium Dodecyl Sulfate Polyacrylamide Gel) was prepared, 1 xTGS (25mM Tris,250mM Glycine,0.1%SDS) running buffer was added and appropriate samples were taken for gel electrophoresis. The initial voltage is 90V, and when the sample enters the lower layer separation gel, the voltage is adjusted to 120V until the electrophoresis is finished.

3. Proteins on the gel were transferred to nitrocellulose membrane using a transfer membrane, and the strips were blocked for 1h at room temperature with a 5% skim milk powder solution (w/v) in TBST solution (10 mM Tris-HCl,150mM NaCl solution containing 1% Tween 20). The bands were washed with distilled water, bands at each target protein were cut, and placed in primary antibody solutions of p-PAK4 (S474), p-beta-Catenin (S675) and beta-action, respectively, and incubated overnight at 4 ℃.

4. The primary antibody was recovered and the strips were washed 3 times with TBST solution for 10min each on a shaker. The strips were incubated with a mouse and rabbit secondary antibody solution formulated with TBST solution for 1h and the TBST solution was washed 3 times. And (5) using a color developing instrument to develop color and develop. The compound is initially screened by a 96-well plate system, the initial screening concentration of the compound is 1 mu mol/L, the compound is diluted by 3 times of gradient, and each concentration is double-multiplexed.

5. Downstream phosphorylation levels were semi-quantitatively analyzed by Image J software.

TABLE 2 inhibition of p-PAK4 and p-beta-Catenin by Compounds S19-S21, S23 and S25

Immunoblotting experiments show that the compounds S19-S21, S23 and S25 can obviously inhibit the levels of p-PAK4 and p-beta-Catenin downstream of PAK4, and the results show that the compounds are novel PAK4 inhibitors and can effectively inhibit downstream signal phosphorylation of PAK 4. Furthermore, the inhibition activity of the compounds on PAK4 downstream signals is obviously superior to KPT-9274.

Experimental example 3: test of the solubility of the hydrochloride salts of Compounds S21 and S23 in Water by high Performance liquid chromatography

The compound S21, S23 and KPT-9274 are dissolved in a small amount of methanol, a proper amount of 4mol/L hydrochloric acid methanol solution is added, the mixture is stirred for one hour, the methanol is removed by screwing, and the mixture is repeated for three times, so that the hydrochloride of the compound S21, S23 and KPT-9274 is prepared.

Weighing a proper amount of compound to be dissolved in 1mL of methanol or water, sequentially diluting 5, 10, 50 and 100 times, sequentially detecting the five concentration solutions by high performance liquid chromatography under the condition of the same sample injection amount, and establishing a linear relation between the concentration and the peak area.

Weighing a proper amount of compound to prepare 1mL of saturated aqueous solution, filtering the solution by a filter membrane, detecting the peak area of the filtrate by high performance liquid chromatography, substituting the peak area into a linear equation, and calculating the solubility of the compound in water.

FIG. 1 is a graph of KPT-9274 (hydrochloride) solution concentration versus peak area.

The solubility of KPT-9274 (hydrochloride) in water was calculated to be <0.002mg/mL.

FIG. 2 is a graph showing the linear relationship between the concentration of compound S21 (hydrochloride) solution and the peak area.

The solubility of compound S21 (hydrochloride) in water was calculated to be 0.252.+ -. 0.014mg/mL.

FIG. 3 is a graph showing the linear relationship between the concentration of compound S23 (hydrochloride) solution and the peak area.

The solubility of compound S23 (hydrochloride) in water was calculated to be 33.98.+ -. 1.36mg/mL.

The compounds S21 and S23 both show excellent water solubility, compared with KPT-9274, the solubility of the two compounds in water is respectively improved by 126 times and 16990 times, and the data show that the compounds S21 and S23 have excellent physicochemical properties and are expected to be used for researching and treating PAK4 related diseases.

Summarizing: the medicine taking PAK4 as the target has good market prospect, but no medicine is marketed at present, and more PAK4 inhibitors with novel structures are required to be developed to meet clinical demands. The invention designs and synthesizes a tricyclic PAK4 inhibitor with novel structure based on the structure of a clinical compound ATG-019 (KPT-9274) of Karyopharms company I phase. The PAK4 inhibitor has good cell proliferation inhibition activity on MIAPaCa-2 cells, has good inhibition effect on PAK4 phosphorylation and downstream beta-catenin phosphorylation, and is a PAK4 inhibitor with a brand new structure. Wherein, the cell proliferation inhibition activity, PAK4 downstream signal inhibition activity and water solubility of part of the compounds are obviously superior to those of phase I clinical compound KPT-9274. Therefore, the compounds described in this patent are expected to be effective PAK4 inhibitors in vivo, providing a new choice for the treatment of tumor patients.

All documents mentioned in this application are incorporated by reference as if each were individually incorporated by reference. Further, it will be appreciated that various changes and modifications may be made by those skilled in the art after reading the above teachings, and such equivalents are intended to fall within the scope of the claims appended hereto.

Claims (10)

1. A compound is characterized in that the compound is a compound shown in a formula I or pharmaceutically acceptable salt, solvate and prodrug thereof,

wherein,

a is selected from the group consisting of:wherein R is ⁰ Selected from the group consisting of: hydrogen, C1-C6 alkyl;

v, W, X, Y are each independently selected from the group consisting of: n, CR ⁴ Wherein R is ⁴ Selected from the group consisting of: hydrogen, halogen, hydroxy, C1-C6 alkoxy, amino, cyano, C1-C6 alkyl;

z is selected from the group consisting of:

R ¹ selected from the group consisting of: hydrogen, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C6 alkoxy containing 1, 2 or 3 members selected from N, O,A 4-8 membered heterocycloalkyl of S, a substituted or unsubstituted C6-C10 aryl, a substituted or unsubstituted 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O, S; wherein the substitution independently refers to substitution with one or more substituents selected from the group consisting of: halogen, hydroxy, cyano, C1-C6 alkoxy, amino, C1-C6 alkyl;

R ² selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O, S, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 5-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O, S, Wherein n is selected from the group consisting of: 0. 1, 2, 3, said substitution independently referring to substitution with one or more substituents selected from the group consisting of: halogen, hydroxy, amino, cyano, mercapto, difluoromethyl, oxo (=o), C1-C6 alkoxy, C1-C6 alkyl, said heterocycloalkyl being selected from the group consisting of: monocyclic, parallel, spiro, bridged rings;

R ³ selected from the group consisting of: hydrogen, amino.

2. The compound of claim 1, wherein a is selected from the group consisting of: wherein R is ⁰ H.

3. A compound according to claim 1,selected from the group consisting of:

R ⁴ Selected from the group consisting of: hydrogen, halogen.

4. The compound of claim 1, wherein R ² Selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 4-8 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O, S, substituted or unsubstituted C3-C8 cycloalkyl,Wherein n is selected from the group consisting of: 0. 1, 2, 3, said substitution independently referring to substitution with one or more substituents selected from the group consisting of: halogen, hydroxy, cyano, mercapto, difluoromethyl, oxo (=o), C1-C6 alkoxy, C1-C6 alkyl, said heterocycloalkyl being selected from the group consisting of: monocyclic, fused, spiro, bridged rings.

5. The compound of claim 1, wherein R ¹ Selected from the group consisting of: hydrogen, halogen, substituted or unsubstituted C6-C10 aryl; wherein the substitution independently refers to substitution with one or more substituents selected from the group consisting of: halogen, C1-C6 alkyl.

6. The compound of claim 1, wherein the compound is selected from the group consisting of:

7. a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of a compound of claim 1.

8. Use of a compound according to claim 1 for the preparation of a medicament for the prevention and/or treatment of a disease selected from the group consisting of: cancer, infectious disease, neurological disease.

9. Use of a compound according to claim 1 for the preparation of a medicament for inhibiting PAK4 kinase activity.

10. Use of a compound according to claim 1 for the preparation of a medicament for the prophylaxis and/or treatment of diseases which are associated with PAK4 kinase activity.