A P-glycoprotein inhibitor and its preparation method and application

Abstract

The present invention discloses compounds represented by formula (I) or (II) or pharmaceutically acceptable salts thereof, as well as methods for preparing and using the compounds. These compounds exhibit significant inhibitory activity against drug-resistant tumor cells with high expression of P-glycoprotein, and have a strong effect in reversing multidrug resistance in tumor cells. Some compounds exhibit significantly better P-glycoprotein inhibitory activity than the third-generation P-glycoprotein inhibitor, talizquidar, and exhibit minimal cytotoxicity. Furthermore, these compounds can selectively inhibit P-glycoprotein in intestinal epithelial cells, potentially improving the bioavailability of anticancer agents when administered orally in combination with anticancer agents that are not readily absorbed in the digestive tract due to the inhibitory effect of intestinal P-glycoprotein.

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to an N-substituted-1,2,3,4-tetrahydrobenzofuran[2,3-C]pyridine P-glycoprotein inhibitor, a preparation method thereof, and an application thereof. Background Art

[0002] Multidrug resistance (MDR) refers to a mechanism in which tumor cells, after becoming resistant to one antitumor drug, also develop cross-resistance to other antitumor drugs with different structures and mechanisms of action.

[0003] The occurrence of multidrug resistance is a major reason for the failure of current tumor chemotherapy, and it is also the most common and difficult problem in tumor treatment.

[0004] Therefore, finding drugs that can reverse MDR to inhibit the development of multidrug resistance has become an urgent problem to be solved in cancer treatment.

[0005] The mechanisms of multidrug resistance in tumors are diverse, involving complex molecular biological underpinnings that are currently underdeveloped. However, overexpression of the transmembrane protein P-glycoprotein (P-gp) in tumor cells is considered a primary driver of multidrug resistance. Overexpressed P-glycoprotein utilizes the energy released by ATP hydrolysis to pump anticancer agents (such as vinca alkaloids, anthracyclines, paclitaxel, and doxorubicin) out of tumor cells, resulting in intracellular concentrations of anticancer drugs below their effective concentrations. This in turn causes tumor cells to develop tolerance to multiple chemotherapeutic agents, leading to MDR. Therefore, the combination of P-glycoprotein inhibitors and anticancer agents is expected to address the problem of multidrug resistance in tumors.

[0006] Since the discovery of the first multidrug resistance reversal agent, verapamil, the research on multidrug resistance reversal agents has gone through three generations.

[0007] The first generation, represented by verapamil and cyclosporine A, has significant cardiovascular and other side effects. Second-generation inhibitors, such as Valspodar and Biricodard, have enhanced activity, but these inhibitors significantly affect the plasma pharmacokinetics of anticancer drugs used in combination with them, limiting their clinical application. Third-generation inhibitors, designed and developed based on structure-activity relationship studies, exhibit superior activity and selectivity, such as Elacridar, Tariquidar, and WK-X-34.

[0008] However, due to the emergence of various side effects, there is still no effective reversal agent used in clinical treatment.

[0009] In addition to cancer cells, P-glycoprotein is also found in many normal human tissues, including the liver, small intestine, kidney, and blood-brain endothelium. In all of these tissues, P-glycoprotein is localized to the secretory compartment. This localization suggests that P-glycoprotein plays a role in limiting the absorption of exogenous toxic substances across biological barriers. For example, anticancer drugs such as paclitaxel are typically administered clinically via intravenous injection due to their physical and chemical properties and the excretion of P-glycoprotein by intestinal epithelial cells.

[0010] Therefore, a new generation of safe and effective P-glycoprotein inhibitors is urgently needed in clinical practice to solve problems such as tumor multidrug resistance and improve the bioavailability of anticancer agents.

[0011] In response to the above problems, the present invention has developed a class of P-glycoprotein inhibitors with novel structures and potent P-glycoprotein inhibitory activity. Summary of the Invention

[0012] The present invention provides compounds represented by formula (I) or (II) or pharmaceutically acceptable salts thereof having P-glycoprotein inhibitory activity, none of which exhibits obvious cytotoxic effects, and some of the compounds or pharmaceutically acceptable salts thereof have P-glycoprotein inhibitory activity significantly superior to that of the third-generation P-glycoprotein inhibitor taliciquidate. The compounds represented by formula (I) or (II) or pharmaceutically acceptable salts thereof have potential for preventing and treating the occurrence of multidrug resistance in tumors, can improve the oral bioavailability of anticancer agents, and have potential anti-tumor value.

[0013] A compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof:

[0014]

[0015] R 1 、R 2 、R 3 、R 4 、R 6 、R 7 are the same or different and are independently selected from H, halogen, C1-C5 alkyl or C1-C5 alkoxy;

[0016] R 5 、R 8 the same or different, each independently selected from substituted or unsubstituted phenyl, pyridyl, pyrazinyl, quinolyl, isoquinolyl, chromonyl, quinolone or quinoxalinyl; the substituents on the phenyl, pyridyl, pyrazinyl, quinolyl, isoquinolyl, chromonyl, quinolone or quinoxalinyl are each independently selected from cyano or halogen;

[0017] m is selected from 0 to 4;

[0018] X is selected from -NHC(=O)- or -NHS(=O)2-;

[0019] L is selected from n is independently selected from 0 to 4.

[0020] The pharmaceutically acceptable salts of the present invention may include addition salts formed with the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, maleic acid, benzenesulfonic acid, succinic acid and salts formed with similar known acceptable acids.

[0021] Furthermore, in the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof, one end of the benzene ring in L is connected to an amide bond.

[0022] Furthermore, the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof is selected from any one of the following:

[0023] N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine, which can be referred to as compound 10a;

[0024] N-(4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine, which can be referred to as compound 10b;

[0025] N-(4-(2-(7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine, which can be referred to as compound 10c;

[0026] N-(4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine, which can be referred to as compound 10d;

[0027] N-(4-(2-(3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine, which can be referred to as compound 10e;

[0028] N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine, which can be referred to as compound 10f;

[0029] N-(4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine, which can be referred to as compound 10g;

[0030] N-(4-(2-(7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine, which can be referred to as compound 10h;

[0031] N-(4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine, which can be referred to as compound 10i;

[0032] N-(4-(2-(3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine, which can be referred to as compound 10j;

[0033] N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13a;

[0034] N-(2-((4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13b;

[0035] N-(2-((4-(2-(7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13c;

[0036] N-(2-((4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13d;

[0037] N-(2-((4-(2-(3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13e;

[0038] 2-(3-cyanobenzamido)-N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxybenzamide, which can be referred to as compound 13f;

[0039] N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)nicotinamide, which can be referred to as compound 13g;

[0040] N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(3-fluorobenzamido)-4,5-dimethoxybenzamide, which can be referred to as compound 13h;

[0041] N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxy-2-(pyridine-3-sulfonyl)benzamide, which can be referred to as compound 13i;

[0042] N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-2-carboxamide, which can be referred to as compound 13j;

[0043] N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-2-oxoethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13k;

[0044] N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-difluorophenyl)quinoline-3-carboxamide, which can be referred to as compound 131;

[0045] N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)phenyl)quinoline-3-carboxamide, which can be referred to as compound 13m;

[0046] N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxopropyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13n;

[0047] (E)-N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide, which can be referred to as compound 13o;

[0048] (E)-N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamoyl)phenyl)quinoline-3-carboxamide, which can be referred to as compound 13p;

[0049] N-(4-(4,5-dimethoxy-2-(quinoline-3-carboxamido)benzamido)phenyl)-6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridine-2(1H)-carboxamide can be referred to as compound 13q.

[0050] The chemical structures of the specific compounds listed above are:

[0051]

[0052] The present invention also provides a method for preparing the compound represented by formula (I) or (II), and the synthetic route is:

[0053]

[0054] Compound 10 is a compound represented by formula (I), wherein Y is

[0055] Compound 13 is a compound represented by formula (II), wherein Y is -NH-Z is -XR 8 ;

[0056] Compound 1 undergoes ortho-Friedel-Crafts acylation under acetic acid / boron trifluoride etherate conditions to obtain compound 2, compound 2 reacts with ethyl bromoacetate in the presence of cesium carbonate as a base to generate compound 3, compound 3 undergoes hydrolysis reaction to obtain compound 4, compound 4 undergoes cyclization reaction to obtain compound 5, compound 5 undergoes a multi-component reaction with benzylamine and formaldehyde aqueous solution in acetic acid solvent to obtain compound 6, compound 6 is debenzylated to obtain compound 7, compound 7 undergoes a nucleophilic substitution reaction with a halide or an amide condensation reaction to obtain compound 8, compound 8 undergoes a nitro reduction reaction to obtain compound 9; compound 9 undergoes a nucleophilic substitution reaction with compound 14 to obtain compound 10; alternatively, compound 9 undergoes an amide condensation reaction with compound 15 to obtain compound 11, compound 11 undergoes a nitro reduction reaction to obtain compound 12, and compound 12 undergoes an amide condensation reaction or a sulfonamide condensation reaction to obtain compound 13.

[0057] The present invention also provides the use of the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof in the preparation of a P-glycoprotein inhibitor. The P-glycoprotein inhibitor can be an oral preparation.

[0058] The compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof can improve or increase the efficacy of anticancer agents, increase the sensitivity of tumors to anticancer agents, and reduce the MDR of tumors to anticancer agents. Specifically, it can improve the effect of cancer treatment by enhancing the cytotoxicity of chemotherapeutic drugs or increasing the net absorption, distribution, metabolism or elimination characteristics of therapeutic drugs.

[0059] Therefore, the present invention also provides the use of the compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof in the preparation of a drug resistance modulator for improving the therapeutic effect of an anticancer drug. The drug resistance modulator can be an oral preparation.

[0060] The anticancer drugs may be drugs that are poorly absorbed in the digestive tract due to the inhibitory effect of intestinal P-glycoprotein, and may include taxanes, vinca alkaloids, anthracyclines, camptothecins, podophyllotoxin, mitoxantrone, actinomycin, colchicine, and the like. Taxanes include paclitaxel and docetaxel. Vinca alkaloids include vincristine and vinblastine. Anthracyclines include daunorubicin and doxorubicin. Camptothecins include topotecan and irinotecan.

[0061] The cancer may be a solid tumor or a hematological malignancy, and may be selected from leukemia, multiple myeloma, and lymphoma. The leukemia may be acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, or chronic myeloid leukemia. The lymphoma may be Hodgkin's lymphoma, non-Hodgkin's lymphoma, mantle cell lymphoma, follicular lymphoma, B-cell lymphoma, T-cell lymphoma, or diffuse large B-cell lymphoma.

[0062] The present invention also provides a pharmaceutical composition comprising a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, and excipients. A pharmaceutically acceptable carrier refers to a carrier that is compatible with the active ingredient in the composition (in some embodiments, can stabilize the active ingredient) and is harmless to the individual being treated. Pharmaceutical carriers and / or excipients can be selected from diluents, fillers, salts, disintegrants, binders, lubricants, glidants, wetting agents, controlled release matrices, colorants, flavorings, buffers, stabilizers, solubilizers, and combinations thereof. The pharmaceutical composition comprising the compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof described herein can be administered in various known ways, such as oral, topical, rectal, parenteral, inhalation, or implantation. Depending on the therapeutic purpose, the pharmaceutical composition can be prepared into various types of dosage unit forms, such as tablets, pills, powders, liquid preparations, suspensions, emulsions, granules, capsules, suppositories, and injections (solutions and suspensions). To form the pharmaceutical composition into tablet form, any excipient known and widely used in the art may be used. For example, carriers such as lactose, sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, and silicic acid; binders such as water, ethanol, propanol, ordinary syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone, and the like; disintegrants such as dry starch, sodium alginate, agar powder, kelp powder, sodium bicarbonate, calcium carbonate, fatty acid esters of polyethylene sorbitan, sodium lauryl sulfate, monoglyceride of stearate, starch, and lactose; disintegration inhibitors such as sugar, glyceryl tristearate, coconut oil, and hydrogenated oil; adsorption promoters such as quaternary ammonium hydroxide and sodium lauryl sulfate; wetting agents such as glycerol and starch; adsorbents such as starch, lactose, kaolin, bentonite, and colloidal silicic acid; and lubricants such as purified talc, stearates, boric acid powder, and polyethylene glycol. Conventional coating materials can also be used to form tablets such as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets, double-layer film tablets, and multi-layer tablets, depending on the needs. To form a pharmaceutical composition in pill form, any known and widely used excipient in the art can be used, including, for example, carriers such as lactose, starch, coconut oil, hardened vegetable oils, kaolin, and talc; binders such as gum arabic powder, tragacanth powder, gelatin, and ethanol; and disintegrants such as agar and kelp powder. To form a pharmaceutical composition in suppository form, any known and widely used excipient in the art can be used, including, for example, polyethylene glycol, coconut oil, higher alcohols, esters of higher alcohols, gelatin, and semi-synthetic glycerides. To prepare a pharmaceutical composition in injectable form, the solution or suspension can be sterilized (preferably by adding an appropriate amount of sodium chloride, glucose, or glycerol) and prepared into an injectable formulation isotonic with blood.When preparing injections, any commonly used carrier in the art may be used, such as water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, and fatty acid esters of polyethylene sorbitan. In addition, common solvents, buffers, and analgesics may be added. In the present invention, there is no particular limitation on the method of administration of the pharmaceutical composition. Various dosage forms may be selected for administration based on the patient's age, gender, and other conditions and symptoms. For example, tablets, pills, solutions, suspensions, emulsions, granules, or capsules may be administered orally; injections may be administered alone or mixed with an injectable delivery fluid (such as a glucose solution or an amino acid solution) for intravenous injection; and suppositories are administered rectally.

[0063] Compared with the prior art, the present invention has the following beneficial effects: the compounds represented by formula (I) or (II) or their pharmaceutically acceptable salts can effectively inhibit P-glycoprotein. These compounds have significant inhibitory activity against drug-resistant tumor cells (such as drug-resistant leukemia cells K562 / A02) that overexpress P-glycoprotein, and have a strong effect in reversing multidrug resistance (MDR) in tumor cells. Some compounds have significantly better P-glycoprotein inhibitory activity than the third-generation P-glycoprotein inhibitor Tariquidar (XR9576) and exhibit less cytotoxicity. Furthermore, these compounds can selectively inhibit P-glycoprotein in intestinal epithelial cells, and when administered orally in combination with anticancer agents (such as paclitaxel, docetaxel, and doxorubicin), which are not easily absorbed in the digestive tract due to the inhibitory effect of intestinal P-glycoprotein, they can improve the bioavailability of the anticancer agents. DETAILED DESCRIPTION

[0064] The present invention will be further described below with reference to specific examples. It should be understood that these examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention.

[0065] Biological Experiment Example 1: Cytotoxic Effects of Compounds

[0066] This experiment tested the cytotoxic effects of 27 compounds on K562 / A02 cells. K562 / A02 cells, a human leukemia cell line resistant to doxorubicin, were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin. Cells in the logarithmic growth phase were plated at a density of 6,000 cells per well in a 96-well plate. Each well was incubated with 100 μL of RPMI 1640 medium at 37°C and 5% CO2 for 24 hours. The doxorubicin control group received 100 μL of a series of doxorubicin concentration gradients per well, while the blank control group received 100 μL of culture medium. The test compound group and the positive control group received 50 μL of the target compound (5 μM) and 50 μL of a series of doxorubicin concentration gradients per well, respectively. Three replicates were set up for each concentration. After addition of the drugs, the plates were incubated for 48 hours, and the solution was discarded. Add 1 mg / mL MTT solution and continue incubation for 4 h. Discard the solution, add DMSO and shake on a shaker for 5 min. Read the plate at a wavelength of 490 nm on a microplate reader to calculate the cell inhibition rate and calculate the compound IC using a dose-effect curve in Graphicpad 5.0 software. 50 value.

[0067] The test results are shown in Table 1. It can be seen from the test results that all compounds have no obvious cytotoxicity to K562 / A02 cells.

[0068] Table 1 Cytotoxic effects of compounds on K562 / A02 cells

[0069]

[0070] Biological Experiment Example 2: Reversal Effect of Compounds on Adriamycin-Resistant Human Leukemia Cells (K562 / A02)

[0071] K562 / A02 cells were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin, using Tariquidar as a positive control. Cells in the logarithmic growth phase were plated at a density of 6,000 cells per well in a 96-well plate. 100 μL of RPMI 1640 medium was added to each well and incubated in a 37°C, 5% CO2 incubator for 24 hours. A doxorubicin control group received 100 μL of a series of doxorubicin concentrations per well, while a blank control group received 100 μL of culture medium. The test compound group and the positive control group received 50 μL of the target compound (5 μM) and 50 μL of a series of doxorubicin concentrations per well, respectively, with triplicate wells set up for each concentration. After drug addition, the cells were incubated for 48 hours, and the solution was discarded. A 1 mg / mL MTT solution was added, and the cells were incubated for another 4 hours. The solution was discarded, and DMSO was added and shaken on a shaker for 5 minutes. The readings were taken at a wavelength of 490 nm on a microplate reader, and the cell inhibition rate was calculated. The IC value of the compound was calculated using the dose-effect curve in Graphicpad 5.0 software. 50value, and find the reversal multiple (RF), the reversal multiple is IC 50 (ADM) / IC 50 (ADM+reversal agent) indicates.

[0072] In this experiment, the reversal activity of 27 compounds against doxorubicin-resistant leukemia cells at a concentration of 5 μM was determined in batches. As shown in Table 2, the test results showed that the vast majority of the 27 compounds had MDR reversal activity, and the reversal activity of some compounds exceeded that of the positive control Tariquidar.

[0073] Table 2 Reversal effect of compounds (5 μM concentration) on drug resistance of doxorubicin-resistant human leukemia cells

[0074]

[0075] Biological Experiment Example 3: Oral Administration of Compound 13o in Combination with Anticancer Agents to Improve the Bioavailability of Anticancer Agents

[0076] Anti-tumor drugs such as paclitaxel (PTX), docetaxel (DTX), and doxorubicin (DOX) are typically administered clinically via intravenous injection due to their physicochemical properties and the excretion of P-glycoprotein by intestinal epithelial cells. Compound 13o and its pharmaceutically acceptable salts in the present invention have a certain inhibitory effect on intestinal P-glycoprotein, potentially improving the oral bioavailability of anticancer drugs.

[0077] Paclitaxel is an important anti-tumor drug that binds to intracellular tubulin, hindering the normal division and proliferation of tumor cells. It is primarily used as a first-line and follow-up treatment for advanced ovarian cancer; adjuvant treatment for patients with node-positive breast cancer following standard doxorubicin-containing combination chemotherapy; breast cancer recurring within 6 months; patients with non-small cell lung cancer; and the treatment of Kaposi's sarcoma. Paclitaxel is typically administered clinically as a solution for intravenous administration, which can be prone to a range of adverse reactions, including peripheral neuropathy, bone marrow suppression, and skin toxicity. Therefore, oral administration of compound 13o and its pharmaceutical salt in combination with paclitaxel can enhance paclitaxel's bioavailability, change the traditional intravenous administration of paclitaxel, mitigate its adverse side effects, and improve patient compliance.

[0078] Six 14-15 week old Sprague-Dawley (SD) rats were fasted for more than 12 hours while allowed to drink water and then randomly divided into two groups. The first group was given 20 mg / kg paclitaxel by gavage; the second group was given 20 mg / kg paclitaxel and 10 mg / kg compound 13o simultaneously by gavage. Blood samples were collected at 0.5, 1, 2, 4, 6, 8, 10, 12, and 24 hours to measure paclitaxel plasma concentrations.

[0079] The test results are shown in Table 3. Last_plasma The results show that compound 13o can significantly improve the oral bioavailability of paclitaxel, which is not easily absorbed in the digestive tract.

[0080] Table 3 Pharmacokinetic parameters of PTX in rats

[0081]

[0082] Note: AUC Last_plasma represents the area under the drug-time curve; C max represents the peak concentration of the drug; T max Indicates the time when the drug peak concentration reaches the peak; T 1 / 2 V represents the drug elimination half-life; Z / F represents the apparent distribution volume of the drug; CL Z / F represents drug clearance rate.

[0083] Preparation Example 1

[0084]

[0085] Preparation of 3,4-dimethoxy-2-hydroxyacetophenone: 20 g of 3,4-dimethoxyphenol was added to a 250 mL two-necked flask under argon. 80 mL of 48% boron trifluoride etherate and 12 mL of acetic acid were added at 0°C. The reaction was continued at this temperature for 30 minutes, then moved to an 80°C oil bath for 3 hours. After the reaction was completed, water was added to quench the mixture at 0°C. A large amount of yellow-green solid precipitated, which was filtered and the filter cake was recrystallized from methanol. Drying under reduced pressure afforded 18.3 g of a gray solid (yield: 72%). 1 H NMR (400MHz, CDCl3) δ12.64(s,1H),7.04(s,1H),6.44(s,1H),3.90(s,3H),3.86(s,3H),2.55(s,3H). 13 CNMR(101MHz, CDCl3)δ202.09,160.14,156.83,141.91,111.74,111.69,100.57,56.71,56.20,26.39.

[0086] Preparation Example 2

[0087]

[0088] Preparation of 2,4-dihydroxyacetophenone: Add 15g of resorcinol to a 250mL two-necked flask, protect with argon, add 80mL of 48% boron trifluoride ether solution and 12mL of acetic acid at 0℃, continue to react at this temperature for 30min, then move to an 80℃ oil bath to react for 3h. After the reaction is completed, quench with water at 0℃, extract with dichloromethane three times (100mL×3), combine the organic phases, wash three times with water, wash once with saturated brine, dry with anhydrous sodium sulfate, and remove the solvent under reduced pressure to obtain a crude product. The crude product is mixed with a small amount of organic solvent (V 乙酸乙酯 :V 石油醚 =1:3) slurry, filtered, and dried under reduced pressure to obtain 16.5 g of a white solid (yield: 80%). 1 H NMR (400MHz, CDCl3) δ12.68 (s, 1H), 7.64 (d, J = 8.5Hz, 1H), 6.46–6.34 (m, 2H), 5.91 (s, 1H), 2.56 (s, 3H). 13 C NMR (101MHz, CDCl3) δ202.77,165.12,162.72,133.09,114.33,107.78,103.51,26.22.

[0089] Preparation Example 3

[0090]

[0091] Preparation of 4-benzyloxy-2-hydroxyacetophenone: Weigh 7.6g of 2,4-dihydroxyacetophenone and 7.9g of potassium carbonate and add them to a 100mL two-necked flask. Under argon protection, add 50mL of acetonitrile and slowly add 6.5mL of benzyl bromide dropwise at room temperature, then heat and reflux for 6h. After the reaction is completed, remove the acetonitrile under reduced pressure, extract with dichloromethane three times (50mL×3), combine the organic phases, wash three times with water, wash once with saturated brine, dry over anhydrous sodium sulfate, and remove the solvent under reduced pressure to obtain a crude product. The crude product is mixed with a small amount of organic solvent (V 乙酸乙酯 :V 石油醚 =1:5) slurry, filtered, and dried under reduced pressure to obtain 10.5 g of a white solid (yield: 87%). 1 H NMR (400MHz, CDCl3) δ12.74 (s, 1H), 7.64 (d, J = 9.0Hz, 1H), 7.45–7.32 (m, 5H), 6.55–6.49 (m, 2H), 5.09 (s, 2H), 2.55 (s, 3H). 13C NMR (101MHz, CDCl3) δ202.62,165.23,135.93,132.40,128.74,128.34,127.56,114.14,108.14,101.94,70.24,26.24.

[0092] Preparation Example 4

[0093]

[0094] Preparation of 3,4-dimethoxy-2-hydroxyacetophenone: Weigh 10g of raw material 2,3,4-trihydroxyacetophenone and 17g of potassium carbonate and add them to a 100mL single-necked bottle, add 50mL of DMF, slowly add 8mL of iodomethane at room temperature, and continue to react at room temperature for 24h. After the reaction is completed, add appropriate amount of water to the reaction solution, extract three times with ethyl acetate (50mL×3), combine the organic phases, wash three times with water, wash once with saturated brine, dry over anhydrous sodium sulfate, and remove the solvent under reduced pressure to obtain a crude product. The crude product is mixed with a small amount of organic solvent (V 乙酸乙酯 :V 石油醚 =1:3) slurry, filtered, and dried under reduced pressure to obtain 8.7 g of a white solid (yield: 75%). 1 H NMR (400MHz, CDCl3) δ12.54(s,1H),7.47(d,J=9.0Hz,1H),6.47(d,J=9.0Hz,1H),3.90(s,3H),3.86(s,3H),2.54(s,3H). 13 C NMR (101MHz, CDCl3) δ203.27,158.53,157.04,136.50,127.05,115.33,102.92,60.66,56.13,26.39.

[0095] Preparation Example 5

[0096]

[0097] Preparation of ethyl 2-(2-acetyl-4,5-dimethoxyphenoxy)acetate: 11 g of 3,4-dimethoxy-2-hydroxyacetophenone and 20 g of cesium carbonate were added to a 250 mL two-necked flask. Under argon, 100 mL of acetone and 7.5 mL of ethyl bromoacetate were added by syringe and allowed to react at 60°C for 3 h. After the reaction, the reaction mixture was removed under reduced pressure, 100 mL of water was added, and the mixture was extracted three times with dichloromethane (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:5) and dried under reduced pressure to yield 10.4 g of a white solid (yield: 66%).1 H NMR (400MHz, CDCl3) δ7.40 (s, 1H), 6.39 (s, 1H), 4.68 (s, 2H), 4.26 (q, J = 7.1Hz, 2H), 3.89 (s, 3H), 3.85 (s, 3H), 2.68 (s, 3H), 1.28 (t, J = 7.1Hz, 3H). 13 C NMR (101MHz, CDCl3) δ197.31,168.27,153.64,153.33,143.84,120.05,112.54,97.50,66.67,61.59,56.23,56.15,32.18,14.17.

[0098] Preparation Example 6

[0099]

[0100] Preparation of ethyl 2-(6-acetyl-2,3-dimethoxyphenoxy)acetate: 8.7 g of the raw materials and 15 g of cesium carbonate were weighed and added to a 250 mL two-necked flask. Under argon, 100 mL of acetone and 7.5 mL of ethyl bromoacetate were added by syringe and allowed to react at 60°C for 3 h. After the reaction, the reaction mixture was removed under reduced pressure and extracted three times with dichloromethane (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:3) and dried under reduced pressure to yield 11 g of a white liquid (yield: 88%). 1 H NMR (400MHz, CDCl3) δ7.50–7.42(m,1H),6.72–6.65(m,1H),4.78–4.71(m,2H),4.24–4. 14(m,2H),3.90–3.83(m,3H),3.82–3.75(m,3H),2.66–2.55(m,3H),1.30–1.16(m,3H). 13 C NMR (101MHz, CDCl3) δ198.05,168.91,157.21,151.75,141.20,126.16,125.51,107.26,69.89,61.05,56.07,31.24,14.10.

[0101] Preparation Example 7

[0102]

[0103] Preparation of ethyl 2-(2-acetyl-5-(benzyloxy)phenoxy)acetate: 10 g of the raw materials and 13.5 g of cesium carbonate were weighed into a 250 mL two-necked flask. Under argon, 100 mL of acetone was added, and 5.0 mL of ethyl bromoacetate was added dropwise via syringe at room temperature. The mixture was then reacted at 60°C for 3 h. After completion of the reaction, the reaction mixture was removed under reduced pressure and extracted three times with dichloromethane (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:3) and dried under reduced pressure at room temperature to yield 12.5 g of a white solid (yield: 92%). 1 H NMR (400MHz, CDCl3) δ7.84(d,J=8.7Hz,1H),7.44–7.30(m,5H),6.63(dd,J=8.7,2.0Hz,1H),6.40(d ,J=2.2Hz,1H),5.08(s,2H),4.66(s,2H),4.26(q,J=7.1Hz,2H),2.67(s,3H),1.30(t,J=7.1Hz,3H). 13 C NMR (101MHz, CDCl3) δ197.66,167.97,163.42,158.97,136.08,132.93,128.7 4,128.34,127.57,121.85,106.91,99.98,70.35,65.55,61.58,32.06,14.16.

[0104] Preparation Example 8

[0105]

[0106] Preparation of 2-(2-acetyl-4,5-dimethoxyphenoxy)acetic acid: 10 g of ethyl 2-(2-acetyl-4,5-dimethoxyphenoxy)acetic acid was weighed and added to a 100 mL single-necked flask. 30 mL of methanol was added dropwise to the mixture under ice-cooling. 25 mL of 2N NaOH aqueous solution was added dropwise. The mixture was allowed to react at room temperature for 6 h. After the reaction, the methanol was removed from the reaction mixture under reduced pressure. 2M HCl aqueous solution was added to the reaction mixture under ice-cooling. The pH of the reaction mixture was adjusted to 2-3. Stirring was continued under ice-cooling for 30 min. A large amount of white solid precipitated. This solid was filtered and dried under reduced pressure to obtain 8.1 g of a white solid product (yield: 90%).

[0107] Preparation Example 9

[0108]

[0109] Preparation of 2-(6-acetyl-2,3-dimethoxyphenoxy)acetic acid: 10 g of the starting material was weighed and added to a 100 mL single-necked flask. 30 mL of methanol was added, and 25 mL of 2N NaOH aqueous solution was added dropwise under an ice bath. The mixture was allowed to react at room temperature for 6 h. After the reaction, the methanol was removed under reduced pressure. 2M HCl aqueous solution was added under an ice bath, and the pH was adjusted to 2-3. Stirring was continued under ice bath for 30 min. A large amount of white solid precipitated, which was filtered and dried under reduced pressure to obtain 7.5 g of a white solid product (yield: 83%).

[0110] Preparation Example 10

[0111]

[0112] Preparation of 2-(2-acetyl-5-(benzyloxy)phenoxy)acetic acid: 12 g of the starting material was weighed and added to a 100 mL single-necked flask. 30 mL of methanol was added, and 25 mL of 2N NaOH aqueous solution was added dropwise under an ice bath. The mixture was allowed to react at room temperature for 6 h. After the reaction, the methanol was removed under reduced pressure. 2 M HCl aqueous solution was added under an ice bath, and the pH of the reaction mixture was adjusted to 2-3. Stirring was continued under ice bath for 30 min. A large amount of white solid precipitated, which was filtered and dried under reduced pressure to yield 10.2 g of a white solid product (93% yield).

[0113] Preparation Example 11

[0114]

[0115] Preparation of 5,6-dimethoxy-3-methylbenzofuran: Add 11 g of 2-(2-acetyl-4,5-dimethoxyphenoxy)acetic acid and 25 g of anhydrous sodium acetate to a 250 mL two-necked flask, add 100 mL of acetic anhydride, and react at 110°C under argon for 12 h. After completion of the reaction, quench with saturated sodium bicarbonate solution in an ice bath until no bubbles form. Extract with ethyl acetate (100 mL x 3). The combined organic phases are washed three times with water and once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product is separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:10) and dried under reduced pressure to yield 6.3 g of a white solid (yield: 76%). 1 H NMR (400MHz, CDCl3) δ7.31(s,1H),7.01(s,1H),6.93(s,1H),3.94(s,3H),3.91(s,3H),2.21(s,3H). 13C NMR (101MHz, CDCl3) δ149.84,147.90,146.31,140.51,120.85,115.67,100.88,95.48,56.47,56.26,8.05.

[0116] Preparation Example 12

[0117]

[0118] Preparation of 6,7-dimethoxy-3-methylbenzofuran: 15 g of the starting material and 34 g of anhydrous sodium acetate were added to a 250 mL two-necked flask. 100 mL of acetic anhydride was added, and the mixture was reacted at 110°C for 12 h under argon. After completion of the reaction, the mixture was quenched with saturated sodium bicarbonate solution under an ice bath until no bubbles formed. The mixture was then extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed three times with water, once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:10) and dried under reduced pressure to yield 9.5 g of a white liquid (yield: 84%). 1 H NMR (400MHz, CDCl3) δ7.32 (d, J = 1.2 Hz, 1H), 7.07 (d, J = 8.4 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 4.14 (s, 3H), 3.90 (s, 3H), 2.18 (d, J = 1.1 Hz, 3H). 13 C NMR (101MHz, CDCl3) δ148.77,147.24,141.02,134.76,125.40,115.60,112.39,109.42,60.85,57.36 7.83.

[0119] Preparation Example 13

[0120]

[0121] Preparation of 6-(Benzyloxy)-3-methylbenzofuran: 15 g of the starting material and 30 g of anhydrous sodium acetate were added to a 250 mL two-necked flask. 100 mL of acetic anhydride was added, and the mixture was reacted at 110°C for 12 h under argon. After completion of the reaction, the mixture was quenched with saturated sodium bicarbonate solution under ice-cooling until no bubbles formed. The mixture was then extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed three times with water, once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:10) and dried under reduced pressure to yield 9.7 g of a white solid (yield: 82%). 1H NMR (400MHz, CDCl3) δ7.50–7.31(m,7H),7.07(d,J=1.7Hz,1H),6.97(dd,J=8.5,1.8Hz,1H),5.11(s,2H),2.22(s,3H). 13 C NMR (101MHz, CDCl3) δ157.09,156.12,140.62,137.06,128.62,127.99,127.53,122.80,119.52,115.52,112.00,97.36,70.66,7.95.

[0122] Preparation Example 14

[0123]

[0124] Preparation of 2-benzyl-6,7-dimethoxy-1,2,3,4-tetrahydrobenzofuran[2,3-c]pyridine: 3.84 g of 5,6-dimethoxy-3-methylbenzofuran was weighed and added to a 100 mL single-necked flask. 40 mL of acetic acid, 32 mL of 36-38% aqueous formaldehyde, and 8.5 g of benzylamine were added. The mixture was reacted at 60°C for 3 h under argon. After the reaction was completed, the mixture was cooled to room temperature and quenched by adding saturated sodium bicarbonate solution until no bubbles formed. The mixture was then extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed three times with water and once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:3) and dried under reduced pressure at room temperature to yield 5.5 g of a white solid (yield: 85%). 1 H NMR (400MHz, CDCl3) δ7.48–7.29(m,5H),7.03(s,1H),6.92(s,1H),3.96(s,3H), 3.94(s,3H),3.81(s,2H),3.68(s,2H),2.90(t,J=5.5Hz,2H),2.77–2.71(m,2H). 13 C NMR (101MHz, CDCl3) δ150.41,149.15,147.04,146.24,138.26,129.11,128.43,1 27.32,120.11,111.35,100.63,95.74,61.88,56.49,56.33,50.34,50.15,20.98.

[0125] Preparation Example 15

[0126]

[0127] Preparation of 2-benzyl-7-(benzyloxy)-1,2,3,4-tetrahydrobenzofuran[2,3-c]pyridine: 7.15 g of 6-(benzyloxy)-3-methylbenzofuran was weighed and added to a 250 mL single-necked bottle. 60 mL of acetic acid, 48 mL of 36-38% aqueous formaldehyde, and 13 g of benzylamine were added. The reaction was carried out under argon at 60°C for 3 h. After completion of the reaction, the mixture was cooled to room temperature and quenched by addition of saturated sodium bicarbonate solution until no bubbles formed. The mixture was then extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:10) and dried under reduced pressure to yield 5.6 g of a white solid (yield: 51%). 1 H NMR (400MHz, CDCl3) δ7.48–7.28(m,11H),7.04(d,J=2.1Hz,1H),6.93(dd,J=8.5,2.2Hz ,1H),5.09(s,2H),3.87(s,2H),3.73(s,2H),2.96(t,J=5.2Hz,2H),2.83–2.72(m,2H). 13 C NMR (101MHz, CDCl3) δ156.60,155.65,137.02,129.44,128.60,127.98,127.82,12 7.53,121.64,118.72,111.85,111.05,97.77,70.72,61.26,49.87,49.66,20.28.

[0128] Preparation Example 16

[0129]

[0130] Preparation of 2-benzyl-7,8-dimethoxy-1,2,3,4-tetrahydrobenzofuran[2,3-c]pyridine: 1.92 g of 6,7-dimethoxy-3-methylbenzofuran was weighed and added to a 100 mL single-necked bottle. 10 mL of acetonitrile, 10 mL of 36-38% aqueous formaldehyde, and 5.7 g of benzylamine hydrochloride were added and reacted at 90°C for 3 h. After the reaction, the mixture was cooled to room temperature and quenched by adding saturated sodium bicarbonate until no bubbles formed. The mixture was then extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:20) and dried under reduced pressure to yield 1.7 g of a white solid (yield: 53%). 1H NMR (400MHz, CDCl3) δ7.42–7.26(m,5H),7.00(d,J=8.4Hz,1H),6.87(d,J=8.4Hz,1H),4.11(s, 3H), 3.91 (s, 3H), 3.78 (s, 2H), 3.67–3.63 (m, 2H), 2.87 (t, J = 5.6Hz, 2H), 2.69 (t, J = 5.6Hz, 2H). 13 C NMR (101MHz, CDCl3) δ151.13,148.43,146.63,138.17,134.81,129.11,128.43,12 7.34,124.43,111.61,111.27,109.20,61.83,60.96,57.33,50.19,50.06,20.86.

[0131] Preparation Example 17

[0132]

[0133] Preparation of 6,7-dimethoxy-1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine: Weigh 3.2 g of 2-benzyl-6,7-dimethoxy-1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine into a 100 mL two-necked flask. Add 640 mg of 10 wt% Pd / C, 20 mL of THF, and 20 mL of methanol. Replace the atmosphere with hydrogen three times and allow the reaction to proceed at room temperature for 12-24 hours. After the reaction is complete, filter (using diatomaceous earth as a filter aid), wash the filter cake with a dichloromethane / methanol mixture, and remove the solvent under reduced pressure to obtain the crude product. The crude product is separated by column chromatography using a mobile phase of methanol:dichloromethane (1:10) and dried under reduced pressure to yield 2.1 g of a white solid (yield: 90%). 1 H NMR (400MHz, DMSO) δ7.24 (d, J = 8.3 Hz, 1H), 6.87 (s, 1H), 6.71 (dd, J = 8.2, 1.4 Hz, 1H), 3.79 (s, 2H), 3.32 (bs, 2H), 2.95 (s, 2H), 2.53 (s, 2H). 13 C NMR (101MHz, CDCl3) δ151.24,148.64,147.11,146.21,120.25,111.69,100.52,95.67,56.46,56.31,43.33,43.24,22.68.

[0134] Preparation Example 18

[0135]

[0136] Preparation of 1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridin-7-ol: 6.0 g of the starting material was weighed into a 100 mL two-necked flask, followed by 1.2 g of 10 wt% Pd / C and 50 mL of ethyl acetate. The mixture was replaced with hydrogen three times and allowed to react at 60°C for 6 h. After the reaction, the mixture was filtered (using diatomaceous earth as a filtration aid), the filter cake was washed with a dichloromethane / methanol mixture, and the solvent was removed under reduced pressure to yield a crude product. The crude product was slurried with a small amount of ethyl acetate, filtered, and dried under reduced pressure to yield 2.5 g of an off-white solid (yield: 81%). 1 H NMR (400MHz, DMSO) δ7.24 (d, J = 8.3 Hz, 1H), 6.87 (s, 1H), 6.71 (dd, J = 8.2, 1.4 Hz, 1H), 3.79 (s, 2H), 2.95 (s, 2H), 2.53 (s, 2H). 13 C NMR (101MHz, DMSO) δ155.25,155.05,151.25,120.68,118.97,111.78,111.71,98.32,42.97,42.68,22.43.

[0137] Preparation Example 19

[0138]

[0139] Preparation of tert-butyl 7-hydroxy-3,4-dihydrobenzofurano[2,3-c]pyridine-2(1H)-carboxylate: 3.8 g of the starting material was weighed and added to a 100 mL single-necked flask. 30 mL of methanol was added, and 4.8 g of Boc2O was slowly added at 0°C. Subsequently, 2.8 mL of triethylamine was slowly added dropwise. The reaction was continued at 0°C for 1 hour. After completion of the reaction, the solvent was removed under reduced pressure, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:3) and dried under reduced pressure to yield 4.5 g of a white solid (yield: 77%). 1 H NMR (400MHz, CDCl3) δ7.22(d,J=8.3Hz,1H),6.95(s,1H),6.80(dd,J=8.3,2.1Hz ,1H),6.54(s,1H),4.55(s,2H),3.73(s,2H),2.66(t,J=5.6Hz,2H),1.51(s,9H). 13 C NMR (101MHz, CDCl3) δ155.68,155.36,153.81,121.09,118.77,111.56,98.61,80.81,42.48,40.93,28.49,20.78.

[0140] Preparation Example 20

[0141]

[0142] Preparation of tert-butyl 7-((trifluoromethylsulfonyl)oxy)-3,4-dihydrobenzofurano[2,3-c]pyridine-2(1H)-carboxylate: 2.9 g of the starting material was weighed into a 50 mL two-necked flask, followed by 25 mL of anhydrous dichloromethane. 2.5 mL of trifluoromethanesulfonic anhydride and 2.0 mL of triethylamine were added dropwise at -5°C, and the reaction was continued for 3 h. After completion, the reaction was quenched with a small amount of water and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:5) and dried under reduced pressure at room temperature to yield 3.5 g of a white solid (yield: 83%). 1 H NMR (400MHz, CDCl3) δ7.46 (d, J = 8.5 Hz, 1H), 7.40 (d, J = 2.2 Hz, 1H), 7.17 (dd, J = 8.5, 2.2 Hz, 1H), 4.61 (s, 2H), 3.76 (s, 2H), 2.73 (s, 2H), 1.50 (s, 9H). 13 C NMR (101MHz, CDCl3) δ154.85,154.04,146.02,128.02,120.40,119.22,117.20,116.30,105.46,80.64,28.40.

[0143] Preparation Example 21

[0144]

[0145] Tert-Butyl 3,4-dihydrobenzofurano[2,3-c]pyridine-2(1H)-carboxylate: 1.26 g of starting material, 33 mg of Pd(OAc)2, and 80 mg of triphenylphosphine were weighed into a 25 mL two-necked flask. 15 mL of DMF was added, and the atmosphere was replaced with argon three times. 0.24 mL of formic acid and 1.2 mL of triethylamine were added via syringe and reacted at 100°C for 3 h. After the reaction, the mixture was cooled to room temperature and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:10) and dried under reduced pressure at room temperature to yield 650 mg of a white solid (yield: 79%). 1H NMR (400MHz, CDCl3) δ7.45–7.38(m,2H),7.26–7.18(m,2H),4.59(s,2H),3.74(s,2H),2.71(s,2H),1.50(s,9H). 13 C NMR (101MHz, CDCl3) δ154.99,154.77,127.83,123.71,122.65,118.69,111.18,80.35,42.47,40.82,28.47,20.83.

[0146] Preparation Example 22

[0147]

[0148] Preparation of 1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine hydrochloride: Weigh 2.7 g of the starting material into a pressure-resistant tube, add 10 mL of hydrogen chloride in ethyl acetate, and react at 50°C for 6 h. After the reaction, remove the solvent under reduced pressure, slurry the residual solid with a small amount of ethyl acetate, filter, and dry under reduced pressure to obtain 1.6 g of a white solid (yield: 77%). 1 H NMR (400MHz, DMSO) δ10.24(s,2H),7.68–7.55(m,2H),7.40–7.26(m,2H),4.39(s,2H),3.53(s,2H),2.96(s,2H). 13 CNMR(101MHz,DMSO)δ154.58,146.07,127.09,125.09,123.66,119.86,111.74,111.68,41.14,40.08,18.01.

[0149] Preparation Example 23

[0150]

[0151] Preparation of 6,7-dimethoxy-2-(4-nitrophenethyl)-1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine: 1.2 g of 6,7-dimethoxy-1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine, 2.3 g of 4-nitrophenethyl bromide, 83 mg of KI, and 1.0 g of triethylamine were weighed and added to a 25 mL two-necked flask. 10 mL of DMF was added, and the mixture was reacted at 100°C under argon for 2 h. After the reaction, the mixture was cooled to room temperature, an appropriate amount of water was added, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:3) and dried under reduced pressure at room temperature to yield 1.1 g of a yellow solid (yield: 58%). 1 H NMR (400MHz, CDCl3) δ8.16(d,J=8.5Hz,2H),7.41(d,J=8.4Hz,2H),7.03(s,1H),6.90(s,1H) ,3.94(s,3H),3.92(s,3H),3.73(s,2H),3.06–2.98(m,2H),2.95–2.87(m,4H),2.73(s,2H). 13 C NMR (101MHz, CDCl3) δ149.86,149.15,148.10,147.15,146.57,146.30,129.56,123. 66,119.91,111.41,100.63,95.72,58.30,56.48,56.33,50.54,50.25,34.08,20.95.

[0152] Preparation Example 24

[0153]

[0154] Preparation of (E)-1-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)-3-(4-nitrophenyl)prop-2-en-1-one: Weigh 690 mg of 6,7-dimethoxy-1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine, 700 mg of cinnamic acid, and 1.35 g of HATU into a two-necked flask. Under argon, 5 mL of DMF and 450 mg of triethylamine were added to the reaction system via syringe at 0°C. The reaction was continued dropwise for 30 minutes, then the reaction was allowed to proceed to room temperature for 1 hour. After the reaction, an appropriate amount of water was added, and the mixture was extracted with DCM (50 mL x 3). The organic phases were combined, washed three times with water, once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography with a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to obtain 1.0 g of a yellow solid (yield: 82%). 1 H NMR (400 MHz, CDCl3, conformer) δ 8.22 (d, J = 8.5 Hz, 2H), 7.78–7.63 (m, 3H), 7.14–6.96 (m, 2H), 6.87 (d, J = 8.5 Hz, 1H), 4.82 (d, J = 16.1 Hz, 2H), 4.12–3.84 (m, 8H), 2.80 (d, J = 18.9 Hz, 2H). 13 C NMR (101 MHz, CDCl3, conformer) δ 165.39, 149.40, 148.19, 148.11, 147.71, 147.59, 146.76, 146.59, 146.52, 141.34, 140.50, 128.42, 124.17, 121.83, 121.57, 119.26, 113.28, 111.36, 100.62, 100.48, 95.78, 95.65, 56.50, 56.34, 44.13, 44.00, 41.50, 40.37, 22.07, 20.60.

[0155] Preparation Example 25

[0156]

[0157] Preparation of tert-butyl (4-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-ylmethyl)phenyl)carbamate: 1.2 g of 6,7-dimethoxy-1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine and 1.3 g of tert-butyl (4-(chloromethyl)phenyl)carbamate were weighed and added to a 25 mL two-necked flask under argon. 10 mL of DMF was added, followed by the slow dropwise addition of 1 mL of triethylamine. The mixture was then heated to 60°C for 2 h. After the reaction, the mixture was cooled to room temperature and extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:3) and dried under reduced pressure at room temperature to yield 1.7 g of a yellow solid (yield: 77%). 1 H NMR (400MHz, CDCl3) δ7.39–7.27(m,4H),6.97(s,1H),6.85(s,1H),6.70(s,1H),3.89(s, 3H),3.87(s,3H),3.72(s,2H),3.63(s,2H),2.89–2.80(m,2H),2.68(s,2H),1.50(s,9H). 13 C NMR (101MHz, CDCl3) δ152.90,149.78,149.17,147.07,146.22,137.78,131.96,129.93,119. 93,118.62,111.31,100.61,95.71,80.52,61.06,56.45,56.30,50.02,49.86,28.37,20.67.

[0158] Preparation Example 26

[0159]

[0160] Preparation of 4-((6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)methyl)aniline: 1.5 g of the starting material was weighed and added to a 50 mL single-necked flask. 10 mL of ethanol and 5 mL of 6N HCl were added and the mixture was allowed to react at 60°C for 1 h. After the reaction, the mixture was cooled to room temperature and the pH was adjusted to 8-9 with 2N NaOH (aq.). The mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed three times with water and once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of ethyl acetate:petroleum ether (1:1) and dried under reduced pressure at room temperature to yield 950 mg of a yellow solid (yield: 83%). 1HNMR (400MHz, CDCl3) δ7.15(d,J=8.2Hz,2H),6.98(s,1H),6.86(s,1H),6.65(d,J=8.2Hz,2H ), 3.90 (s, 3H), 3.88 (s, 3H), 3.78–3.49 (m, 6H), 2.82 (t, J = 5.6Hz, 2H), 2.67 (t, J = 5.3Hz, 2H). 13 C NMR (101MHz, CDCl3) δ150.55,149.12,146.96,146.17,145.71,130.35,127.84,1 20.13,115.04,111.37,100.62,95.73,61.45,56.47,56.31,50.13,49.92,20.96.

[0161] Preparation Example 27

[0162]

[0163] Preparation of 4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)aniline: Weigh 1.0 g of 6,7-dimethoxy-2-(4-nitrophenylethyl)-1,2,3,4-tetrahydrobenzofurano[2,3-c]pyridine into a 25 mL two-necked flask, add 100 mg of 10 wt% Pd / C and 10 mL of methanol, and react at room temperature for 2 h. After the reaction is complete, filter, and remove the solvent under reduced pressure to obtain the crude product. The crude product is separated by column chromatography with a mobile phase of ethyl acetate:petroleum ether (1:1) and dried under reduced pressure at room temperature to obtain 810 mg of a yellow solid (yield: 88%). 1 H NMR (400MHz, CDCl3) δ7.05–6.99(m,3H),6.88(s,1H),6.62(d,J=8.3Hz,2H),3.92(s,3H),3.9 0(s,3H),3.71(s,2H),3.57(s,2H),2.89(t,J=5.6Hz,2H),2.81(s,4H),2.72(t,J=5.4Hz,2H). 13 C NMR (101MHz, CDCl3) δ150.29,149.16,147.04,146.22,144.64,129.98,129.52,120. 06,115.33,111.41,100.65,95.75,59.89,56.48,56.33,50.63,50.34,33.39,21.03.

[0164] Preparation Example 28

[0165]

[0166] (E)-3-(4-Aminophenyl)-1-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)prop-2-en-1-one: 610 mg of (E)-1-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-(4-nitrophenyl)prop-2-en-1-one and 900 mg of stannous chloride were weighed and added to a pressure tube. 6 mL of ethanol and 6 mL of 6N HCl were added, and the mixture was reacted at 90°C for 3 h. After completion of the reaction, the mixture was extracted with DCM (50 mL × 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain the crude product. The crude product was separated by column chromatography with a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to obtain 450 mg of a yellow solid (yield: 79%). 1 H NMR (400MHz, CDCl3) δ7.65(d,J=15.3Hz,1H),7.37(d,J=8.3Hz,2H),7.02(s,1H),6.88(s,1H) ,6.76(d,J=14.6Hz,1H),6.65(d,J=8.4Hz,2H),4.82(s,2H),4.06–3.88(m,10H),2.77(s,2H). 13 C NMR (101MHz, CDCl3) δ167.03,149.34,148.31,147.45,146.44,143.75,129.61,125.45,119.52,114.89,100.56,95.74,56.49,56.34.

[0167] Preparation Example 29

[0168]

[0169] Preparation of (E)-N-(4-(3-(7,8-dimethoxy-1,2,3,4-tetrahydrodibenzo[b,d]furan-3-yl)-3-oxoprop-1-en-1-yl)phenyl)-4,5-dimethoxyoxy-2-nitrobenzamide: Weigh 340 mg of 4,5-dimethoxy-2-nitrobenzoic acid and add it to a two-necked flask. Under argon protection, add 5 mL of anhydrous dichloromethane and 1 drop of DMF via syringe. At 0°C, slowly add 0.4 mL of oxalyl chloride via syringe to the reaction system. Continue the reaction for 30 minutes and then move to room temperature for 3 hours. After the reaction, the solvent and excess oxalyl chloride were removed under reduced pressure. The residual solid was dissolved in 3 mL of anhydrous dichloromethane and slowly added to a pre-prepared reaction mixture of 380 mg of (E)-3-(4-aminophenyl)-1-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)prop-2-en-1-one in 5 mL of DCM. 200 mg of triethylamine was then added, and the reaction continued at room temperature for 1 hour. After the reaction, an appropriate amount of water was added, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to yield 440 mg of a pale yellow solid (yield: 75%). 1H NMR (400MHz, CDCl3) δ8.95–8.49(m,1H),7.63(d,J=7.1Hz,2H),7.59–7.38(m,4H),7.01(s,1H),6. 95(d,J=9.2Hz,1H),6.91–6.75(m,2H),4.81–4.68(m,2H),4.00–3.82(m,14H),2.65-2.85(m,2H).

[0170] Preparation Example 30

[0171]

[0172] Preparation of N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxy-2-nitrobenzamide: Weigh 340 mg of 4,5-dimethoxy-2-nitrobenzoic acid and add it to a two-necked flask. Under argon protection, add 5 mL of anhydrous dichloromethane and 1 drop of DMF via syringe. At 0°C, slowly add 0.4 mL of oxalyl chloride via syringe to the reaction system. Continue the reaction for 30 minutes and then move to room temperature for 3 hours. After the reaction, the solvent and excess oxalyl chloride were removed under reduced pressure. The residual solid was dissolved in 3 mL of anhydrous dichloromethane and slowly added to a pre-prepared reaction mixture of 350 mg of 4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)aniline in 5 mL of DCM. 200 mg of triethylamine was then added, and the reaction continued at room temperature for 1 hour. After the reaction, an appropriate amount of water was added, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to yield the crude product. The crude product was separated by column chromatography using a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to yield 410 mg of a yellow solid (yield: 73%). 1 H NMR (400MHz, CDCl3) δ8.18 (s, 1H), 7.50–7.42 (m, 3H), 7.17 (d, J = 8.3Hz, 2H), 6.96 (s, 1H), 6 .90(s,1H),6.84(s,1H),3.93–3.83(m,12H),3.70(s,2H),2.95–2.80(m,6H),2.70(s,2H). 13 C NMR (101MHz, CDCl3) δ164.85,153.51,150.03,149.33,149.13,147.01,146.16,138.34,136.70,135.87,129.26,127.28, 120.61,119.96,111.41,110.11,107.05,100.64,95.70,59.34,56.68,56.45,56.44,56.28,50.62,50.26,33.61,20.95.

[0173] Preparation Example 31

[0174]

[0175] Preparation of (E)-2-amino-N-(4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxoprop-1-en-1-yl)phenyl)-4,5-dimethoxybenzamide: Weigh 590 mg of the starting material and 300 mg of stannous chloride, add 3 mL of EtOH and 3 mL of 6N HCl, and react at 90°C for 3 h. After completion of the reaction, extract with DCM (50 mL x 3). The combined organic phases are washed three times with water and once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to obtain the crude product. The crude product is separated by column chromatography with a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to obtain 410 mg of a yellow solid (yield: 74%). 1 H NMR (400 MHz, CDCl3+CD3OD, conformer) δ7.63–7.52 (m, 3H), 7.51–7.42 (m, 2H), 7.10–7.01 (m, 1H), 6.96 (s, 1H), 6.90–6.74 (m, 2H), 6.22–6.15 (m, 1H), 4.74 (s, 2H), 4.02–3.67 (m, 14H), 2.81–2.61 (m, 2H). 13 C NMR (101 MHz, CDCl3+CD3OD, conformer) δ 167.71, 167.03, 153.67, 149.35, 147.38, 146.34, 145.17, 143.21, 140.82, 140.17, 130.40, 128.63, 120.64, 119.39, 115.35, 111.87, 107.54, 100.89, 100.61, 95.75, 57.00, 56.41, 56.23, 55.68, 43.94, 41.38, 40.26, 21.88, 20.54.

[0176] Preparation Example 32

[0177]

[0178] Preparation of 2-amino-N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxybenzamide: 400 mg of 2-amino-N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxybenzamide was weighed and added to a 25 mL two-necked flask. 80 mg of 10 wt% Pd / C, 5 mL of methanol, and 5 mL of tetrahydrofuran were added and allowed to react at room temperature for 2 h. After the reaction was complete, the mixture was filtered and the solvent removed under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to obtain 320 mg of a yellow solid (yield: 85%). 1 H NMR (400MHz, CDCl3) δ7.91(s,1H),7.46(d,J=8.4Hz,2H),7.20(d,J=8.4Hz,2H),7.00(s,1H),6.98(s,1H),6.87(s,1H),6.2 0(s,1H),3.92–3.89(m,5H),3.89(s,3H),3.84(s,3H),3.80(s,3H),3.73(s,2H),2.93–2.83(m,6H),2.72(t,J=5.3Hz,2H). 13 C NMR (101MHz, CDCl3) δ167.20,153.64,150.00,149.17,147.06,146.22,144.98,141.08,136.25,135.99,129.23,120.94, 119.97,111.40,111.27,107.64,101.04,100.62,95.72,59.33,57.11,56.45,56.31,55.81,50.56,50.21,33.59,20.91.

[0179] Preparation Example 33

[0180]

[0181] Preparation of N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Weigh 70 mg of quinoline-3-carboxylic acid into a 10 mL single-necked flask, add 5 mL of anhydrous tetrahydrofuran and 0.15 mL of thionyl chloride, and react at 60°C for 12 h under argon protection. After completion of the reaction, remove the solvent and excess thionyl chloride under reduced pressure to obtain a gray solid. 106 mg of 2-amino-N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxybenzamide was weighed and added to a 10 mL single-necked flask. 5 mL of anhydrous dichloromethane was added, and the prepared acid chloride was added to the reaction system. 40 mg of triethylamine was then added, and the reaction continued at room temperature for 1 hour. After the reaction, an appropriate amount of 1N NaOH solution was added, and the mixture was stirred at room temperature for 30 minutes. Extraction was performed with dichloromethane (50 mL x 3). The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to yield 85 mg of compound 13a as a yellow solid (yield: 62%). 1 H NMR (400MHz, CDCl3+CD3OD) δ9.47(s,1H),8.82(s,1H),8.52(s,1H),8.14(d,J=8.3Hz ,1H),8.05(d,J=8.1Hz,1H),7.85(t,J=7.4Hz,1H),7.67(t,J=7.4Hz,1H),7.59(d,J=7 .9Hz,2H),7.39(s,1H),7.27(d,J=8.0Hz,2H),7.03(s,1H),6.91(s,1H),4.02(s,3H) ,3.96(s,3H),3.92(s,3H),3.91(s,3H),3.76(s,2H),3.00–2.85(m,6H),2.77(s,2H). 13C NMR (101MHz, CDCl3+CD3OD) δ167.90,163.69,152.36,149.41,149.17,148.72,148. 56,147.05,146.12,144.51,136.31,136.01,135.85,135.11,131.72,129.19,129. 05,128.50,127.63,127.29,126.96,121.84,119.75,112.59,111.27,110.75,104.79,100.70,95.70,59.14,56.33,56.29,56.16,55.97,50.42,50.05,33.18,20.52.

[0182] Preparation Example 34

[0183]

[0184] Preparation of N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine: Weigh 105 mg of 4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)aniline and 72 mg of 4-chloro-2-(pyridin-3-yl)quinazoline into a Schlenk tube. Under argon protection, add 3 mL of anhydrous ethanol and 3 μL of methanesulfonic acid, and react at 70°C for 5 h. After the reaction, extract with DCM (30 mL × 3), combine the organic phases, wash three times with water, wash once with saturated brine, dry over anhydrous sodium sulfate, and remove the solvent under reduced pressure to obtain the crude product. The crude product was separated by column chromatography with a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to obtain 110 mg of compound 10a as a yellow solid (yield: 66%). 1H NMR (500MHz, CDCl3) δ9.73 (d, J=1.6Hz, 1H), 8.79 (dt, J=7.9, 1.8Hz, 1H), 8.69 (dd, J=4.7, 1.5Hz,1H),7.99(d,J=8.3Hz,1H),7.91(d,J=8.2Hz,1H),7.85–7.77(m,3H),7.60(s,1H),7 .55(t,J=7.3Hz,1H),7.41(dd,J=7.8,4.8Hz,1H),7.34(d,J=8.3Hz,2H),7.02(s,1H),6.8 9(s,1H),3.92(s,3H),3.91(s,3H),3.77(s,2H),3.00–2.89(m,6H),2.75(t,J=5.2Hz,2H). 13 C NMR (126MHz, CDCl3) δ158.51,157.49,150.83,150.30,150.11,149.15,147.04,146.21,136.46,136.24,135.75,134.18,133.10,1 29.31,126.52,123.27,121.69,120.36,120.00,114.02,111.41,100.62,95.72,59.36,56.46,56.31,50.63,50.33,33.73,20.99.

[0185] Preparation Example 35

[0186]

[0187] Preparation of N-(4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine: Weigh 100 mg of 4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)aniline and 72 mg of 4-chloro-2-(pyridin-3-yl)quinazoline into a Schlenk tube. Under argon protection, add 3 mL of anhydrous ethanol and 3 μL of methanesulfonic acid, and react at 70°C for 5 h. After the reaction is completed, extract with DCM (30 mL × 3), combine the organic phases, wash three times with water, wash once with saturated brine, dry over anhydrous sodium sulfate, and remove the solvent under reduced pressure to obtain the crude product. The crude product was separated by column chromatography with a mobile phase of dichloromethane:methanol = 50:1 and dried under reduced pressure at room temperature to obtain 95 mg of compound 10b as a yellow solid (yield: 58%). 1H NMR(500MHz, CDCl3)δ9.74(s,1H),8.79(s,1H),8.69(s,1H),8.11–7.75(m,5H),7.73–7.33(m,5H),7. 00(s,1H),6.88(s,1H),3.92(s,3H),3.90(s,3H),3.81(s,2H),3.69(s,2H),2.92(s,2H),2.73(s,2H). 13 C NMR (126MHz, CDCl3) δ158.52,157.46,150.87,150.29,149.14,147.02,146.21,137.52,135.77,134.25,134.16,133.16,129.8 0,129.38,126.60,123.30,121.36,120.35,120.06,114.04,111.37,100.60,95.71,61.33,56.46,56.31,50.32,50.14,20.96.

[0188] Preparation Example 36

[0189] Preparation of N-(4-(2-(7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine:

[0190] Referring to the preparation method in Preparation Example 34, yellow solid compound 10c was prepared (yield: 68%). 1 HNMR(500MHz, CDCl3)δ9.74–9.71(m,1H),8.78(d,J=7.9Hz,1H),8.71–8.66(m,1H),7.97(dd,J =8.3,1.4Hz,1H),7.93(d,J=8.3Hz,1H),7.83–7.76(m,3H),7.74–7.63(m,1H),7.53(dd,J=11. 8,7.4Hz,1H),7.42–7.38(m,1H),7.32(dd,J=8.2,2.7Hz,2H),7.01(d,J=8.3Hz,1H),6.87(d,J =8.3Hz,1H),4.13(s,3H),3.91(s,3H),3.77(d,J=1.1Hz,2H),2.98–2.88(m,6H),2.73(s,2H). 13C NMR (126MHz, CDCl3) δ158.50,157.53,150.90,150.84,150.83,150.31,148.50,146.71,136.51,136.20,135.76,134.79,134.21,133.0 8,129.28,126.51,124.33,123.28,121.73,120.48,114.05,111.71,111.37,109.23,61.01,59.29,57.31,50.49,50.23,33.73,20.88.

[0191] Preparation Example 37

[0192] Preparation of N-(4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine: Referring to the preparation method in Preparation Example 35, a yellow solid compound 10d was prepared (yield: 63%). 1 H NMR(500MHz, CDCl3)δ9.75(s,1H),8.79(dt,J=7.9,1.8Hz,1H),8.70(d,J=3.8Hz,1H),8.00(d,J= 8.3Hz,1H),7.93(d,J=8.1Hz,1H),7.89–7.80(m,3H),7.64(d,J=3.9Hz,1H),7.56(t,J=7.6Hz,1H) ,7.49(d,J=8.3Hz,2H),7.42(dd,J=7.8,4.8Hz,1H),7.01(d,J=8.4Hz,1H),6.87(d,J=8.4Hz,1H), 4.11(s,3H),3.91(s,3H),3.81(s,2H),3.70(s,2H),2.91(t,J=5.6Hz,2H),2.72(t,J=5.4Hz,2H). 13 C NMR (126MHz, CDCl3) δ158.54,157.46,151.06,150.89,150.88,150.30,148.42,146.64,137.52,135.77,134.80,134.24,134.16,133.1 6,129.78,129.39,126.60,124.41,123.30,121.37,120.33,114.03,111.64,111.31,109.21,61.32,60.95,57.32,50.21,50.06,20.88.

[0193] Preparation Example 38

[0194] Prepared by N-(4-(2-(3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine; referring to the preparation method in Preparation Example 34, a yellow solid compound 10e was obtained (yield: 61%). 1 H NMR (500MHz, CDCl3) δ9.73(s,1H),8.79(d,J=7.9Hz,1H),8.69(d,J=3.4Hz,1H),7.99(d,J=8.3Hz,1H),7.91(d,J=8.1Hz,1H),7.85–7.77(m, 3H),7.56(t,J=7.2Hz,2H),7.47–7.39(m,3H),7.35(d,J=8.2Hz,2H),7.25–7.17(m,2H),3.80(s,2H),3.02–2.90(m,6H),2.83–2.76(m,2H). 13 C NMR (126MHz, CDCl3) δ158.55,157.49,154.75,151.26,150.87,150.34,136.45,136.28,135.75,134.17,133.12,129.37,129.34 ,128.09,126.54,123.33,123.27,122.43,121.70,120.33,118.62,114.01,111.43,111.08,59.37,50.58,50.24,33.72,20.88.

[0195] Preparation Example 39

[0196] Preparation of N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine:

[0197] Referring to the preparation method in Preparation Example 34, yellow solid compound 10f was prepared (yield: 63%). 1HNMR(500MHz, CDCl3)δ8.76(d,J=4.0Hz,2H),8.37(d,J=4.6Hz,2H),8.01(d,J= 8.2Hz,1H),7.93(d,J=8.1Hz,1H),7.84(t,J=7.6Hz,1H),7.79(d,J=7.9Hz,2H) ,7.62(s,1H),7.58(t,J=7.4Hz,1H),7.36(d,J=7.9Hz,2H),7.02(s,1H),6.89( s,1H),3.92(s,3H),3.91(s,3H),3.77(s,2H),3.03–2.91(m,6H),2.75(s,2H). 13 C NMR (126MHz, CDCl3) δ158.32,157.64,150.77,150.24,150.08,149.17,147.09,146.25,146.14,136.45,136.35,133.19,129.60,1 29.32,127.01,122.40,121.68,120.37,119.98,114.29,111.42,100.62,95.73,59.34,56.47,56.32,50.67,50.33,33.74,20.99.

[0198] Preparation Example 40

[0199] Preparation of N-(4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine:

[0200] By referring to the preparation method in Preparation Example 35, 10 g of a yellow solid compound was obtained (yield: 65%). 1 HNMR(500MHz, CDCl3)δ8.77(d,J=5.6Hz,2H),8.37(d,J=5.9Hz,2H),8.02(d, J=8.2Hz,1H),7.95(d,J=8.2Hz,1H),7.90–7.82(m,3H),7.67(s,1H),7.59(t ,J=7.4Hz,1H),7.52(d,J=8.3Hz,2H),7.00(s,1H),6.88(s,1H),3.92(s,3H) ,3.90(s,3H),3.83(s,2H),3.69(s,2H),2.93(t,J=5.5Hz,2H),2.74(s,2H). 13C NMR (126MHz, CDCl3) δ158.32,157.62,150.79,150.27,149.15,147.06,146.24,146.09,137.48,134.40,133.24,129.77,1 29.64,127.07,122.39,121.41,120.36,120.04,114.30,111.37,100.60,95.71,61.36,56.47,56.31,50.32,50.23,20.98.

[0201] Preparation Example 41

[0202] Preparation of N-(4-(2-(7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine: Referring to the preparation method in Preparation Example 34, a yellow solid compound 10h was prepared (yield: 63%). 1 H NMR (500MHz, CDCl3) δ8.77 (d, J = 5.6 Hz, 2H), 8.37 (dd, J = 4.7, 1.3 Hz, 2H), 8.02 (d, J = 8. 1Hz,1H),7.93(d,J=8.2Hz,1H),7.86–7.82(m,1H),7.80(d,J=8.4Hz,2H),7.63(s,1H) ,7.58(t,J=7.2Hz,1H),7.36(d,J=8.4Hz,2H),7.02(d,J=8.4Hz,1H),6.88(d,J=8.4Hz ,1H),4.13(s,3H),3.91(s,3H),3.79(s,2H),3.01–2.92(m,6H),2.74(t,J=5.4Hz,2H). 13 C NMR (126MHz, CDCl3) δ158.32,157.65,150.86,150.76,150.24,148.55,146.73,146.15,136.46,136.33,134.80,133.19,129.59,1 29.31,127.01,124.29,122.40,121.67,120.38,114.30,111.72,111.37,109.26,61.01,59.25,57.31,50.52,50.22,33.76,20.87.

[0203] Preparation Example 42

[0204] Preparation of N-(4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine: Referring to the preparation method in Preparation Example 35, a yellow solid compound 10i was prepared (yield: 59%). 1 H NMR (500MHz, CDCl3) δ8.77(d,J=5.6Hz,2H),8.38(d,J=5.8Hz,2H),8.02(d,J=8. 3Hz,1H),7.95(d,J=8.2Hz,1H),7.91–7.81(m,3H),7.70(s,1H),7.59(t,J=7.5Hz ,1H),7.51(d,J=8.2Hz,2H),7.01(d,J=8.4Hz,1H),6.87(d,J=8.4Hz,1H),4.11(s ,3H),3.91(s,3H),3.83(s,2H),3.71(s,2H),2.93(t,J=5.5Hz,2H),2.73(s,2H). 13 C NMR (126MHz, CDCl3) δ158.31,157.61,151.01,150.78,150.26,148.48,146.67,146.11,137.52,134.79,134.32,133.23,129.7 6,129.62,127.07,124.36,122.40,121.37,120.38,114.31,111.68,111.31,109.24,61.30,60.96,57.30,50.20,50.11,20.87.

[0205] Preparation Example 43

[0206] Preparation of N-(4-(2-(3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine: Referring to the preparation method in Preparation Example 34, a yellow solid compound 10j was prepared (yield: 71%). 1H NMR (500MHz, CDCl3) δ8.77(dd,J=4.5,1.6Hz,2H),8.37(dd,J=4.5,1.6Hz,2H),8.02(dd,J=8.4,0.7Hz,1H),7.92(d,J=8.0Hz,1H),7.87–7.82(m,1H),7. 82–7.77(m,2H),7.61–7.56(m,2H),7.47–7.41(m,2H),7.37(d,J=8.4Hz,2H ),7.25–7.19(m,2H),3.81(s,2H),3.12–2.89(m,6H),2.79(t,J=5.6Hz,2H).

[0207] 13 C NMR (126MHz, CDCl3) δ158.33,157.61,154.73,151.22,150.76,150.24,146.11,136.42,136.36,133.18,129.61,129.32,12 8.06,127.01,123.35,122.45,122.38,121.65,120.32,118.60,114.27,111.42,111.07,59.32,50.60,50.22,33.74,20.87.

[0208] Preparation Example 44

[0209] Preparation of N-(2-((4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13b was prepared (yield: 53%). 1H NMR (500MHz, CDCl3) δ12.45(s,1H),9.54(d,J=2.2Hz,1H),8.77(d,J=1.9Hz,1H),8.60(s,1H) ,8.17(d,J=8.4Hz,1H),8.08(s,1H),8.00(d,J=7.9Hz,1H),7.86–7.80(m,1H),7.67–7.61(m, 3H),7.45(d,J=8.3Hz,2H),7.10(s,1H),6.99(s,1H),6.87(s,1H),3.98(s,3H),3.92(s,3H), 3.90(s,3H),3.85(s,3H),3.79(s,2H),3.65(s,2H),2.88(t,J=5.5Hz,2H),2.73–2.69(m,2H). 13 C NMR (101MHz, CDCl3) δ167.49,163.96,152.92,150.19,149.42,149.15,148 .82,147.05,146.22,144.68,136.54,135.85,135.80,131.50,129.90,129. 41,129.24,127.52,127.25,126.95,121.01,120.02,112.14,111.36,109.66,105.06,100.58,95.71,61.25,56.46,56.31,56.19,50.27,50.10,20.94.

[0210] Preparation Example 45

[0211] Preparation of N-(2-((4-(2-(7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13c was prepared (yield: 49%). 1H NMR (500MHz, CDCl3) δ12.45(s,1H),9.53(d,J=2.2Hz,1H),8.76(d,J=1.9Hz,1H),8.58(s,1H),8.17( d,J=8.4Hz,1H),8.08(s,1H),8.00(d,J=8.0Hz,1H),7.87–7.79(m,1H),7.63(t,J=7.4Hz,1H),7.58( d,J=8.4Hz,2H),7.29(d,J=8.3Hz,2H),7.09(s,1H),7.01(d,J=8.4Hz,1H),6.88(d,J=8.4Hz,1H),4. 13(s,3H),3.97(s,3H),3.91(s,3H),3.84(s,3H),3.76(s,2H),2.96–2.86(m,6H),2.75–2.70(m,2H). 13 C NMR (126MHz, CDCl3) δ167.43,163.92,152.90,150.84,149.40,148.81,148.53,1 46.71,144.67,137.14,135.81,135.80,135.43,134.78,131.47,129.44,129.39, 129.20,127.48,127.26,126.94,124.28,121.27,112.13,111.71,111.35,109.64,109.24,105.08,61.00,59.21,57.30,56.50,56.18,50.48,50.19,33.73,20.87.

[0212] Preparation Example 46

[0213] Preparation of N-(2-((4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13d was obtained (yield: 43%). 1H NMR (500MHz, CDCl3) δ12.46 (s, 1H), 9.54 (d, J = 1.8Hz, 1H), 8.78 (s, 1H), 8.62 (s, 1H), 8.17 ( d,J=8.4Hz,1H),8.04–7.97(m,2H),7.83(t,J=7.4Hz,1H),7.67–7.60(m,3H),7.45(d,J=8. 2Hz,2H),7.10(s,1H),7.00(d,J=8.4Hz,1H),6.87(d,J=8.4Hz,1H),4.10(s,3H),4.00(s,3 H), 3.91 (s, 3H), 3.88 (s, 3H), 3.79 (s, 2H), 3.67 (s, 2H), 2.88 (t, J = 5.3Hz, 2H), 2.70 (s, 2H). 13C NMR (101MHz, CDCl3+CD3OD) δ167.80,163.76,152.56,150.61,148.95,148.67,14 8.42,146.60,144.55,136.95,136.01,135.31,134.66,134.17,131.70,129.89,1 29.23,128.82,127.63,127.28,126.98,124.21,121.35,112.50,111.76,111.25,110.53,109.25,104.88,61.20,60.89,57.25,56.41,56.10,49.98,49.89,20.58.

[0214] Preparation Example 47

[0215] Preparation of N-(2-((4-(2-(3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13e was prepared (yield: 47%). 1H NMR (500MHz, CDCl3) δ12.47(s,1H),9.54(d,J=2.3Hz,1H),8.77(d,J=2.1Hz,1H),8.61(s,1H ),8.17(d,J=8.6Hz,1H),8.00(d,J=7.7Hz,1H),7.94(s,1H),7.85–7.80(m,1H),7.65–7.61( m,1H),7.56(d,J=8.4Hz,2H),7.45–7.40(m,2H),7.30(d,J=8.4Hz,2H),7.25–7.19(m,2H),7 .09(s,1H),4.00(s,3H),3.89(s,3H),3.77(s,2H),2.98–2.87(m,6H),2.77(t,J=5.6Hz,2H). 13 C NMR (101MHz, CDCl3) δ167.46,163.92,154.74,152.84,151.13,149.38,148.8 1,144.65,137.06,135.83,135.74,135.51,131.49,129.44,129.38,129.22,1 28.04,127.50,127.25,126.94,123.38,122.47,121.29,118.63,112.19,111.41,111.09,109.71,105.06,59.26,56.47,56.18,50.54,50.17,33.67,20.83.

[0216] Preparation Example 48

[0217] Preparation of 2-(3-cyanobenzamido)-N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxybenzamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13f was prepared (yield: 42%). 1H NMR(500MHz,DMSO)δ12.09(s,1H),10.47(s,1H),8.29(s,1H),8.19(d,J=8.0Hz,1H),8 .15(s,1H),8.09(d,J=7.7Hz,1H),7.80(t,J=7.8Hz,1H),7.67(d,J=8.3Hz,2H),7.53( s,1H),7.33(d,J=8.4Hz,2H),7.30(s,1H),7.17(s,1H),4.76–4.40(m,2H),3.90(s,3H ),3.87(s,3H),3.81(s,6H),3.57–3.39(m,4H),3.24–3.13(m,2H),3.12–2.93(m,2H). 13 C NMR (126MHz, DMSO) δ167.46,163.14,151.92,149.49,148.30,146.95,144.92,137.67,136.28,135.76,133.88,131.97,131.30,130.77,129. 40,122.36,118.71,118.58,115.00,112.53,112.48,111.68,105.79,1 01.97,96.57,56.51,56.49,56.46,56.12,49.95,48.08,29.64,18.26.

[0218] Preparation Example 49

[0219] Preparation of N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)nicotinamide: Referring to the preparation method in Preparation Example 33, 13 g of a pale yellow solid compound was obtained (yield: 45%). 1H NMR (400MHz, CDCl3) δ12.26(s,1H),9.27(d,J=1.7Hz,1H),8.76(dd,J=4.7,1.2Hz,1H ),8.51(s,1H),8.25(d,J=8.0Hz,1H),8.09(s,1H),7.55(d,J=8.3Hz,2H),7.43(dd,J =7.8,4.9Hz,1H),7.29(d,J=8.3Hz,2H),7.05(s,1H),7.01(s,1H),6.88(s,1H),3.92 (s,6H),3.90(s,3H),3.81(s,3H),3.74(s,2H),2.97–2.84(m,6H),2.76–2.70(m,2H). 13 C NMR (101MHz, CDCl3) δ167.43,163.83,152.81,152.58,150.10,149.14,147.01,146.18,144.63,137.22,135.59,135.38,134.81,130.25, 129.49,123.57,121.32,119.95,112.13,111.42,109.51,104.97,10 0.51,95.65,59.37,56.43,56.31,56.19,50.66,50.34,33.77,21.04.

[0220] Preparation Example 50

[0221] Preparation of N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(3-fluorobenzamide)-4,5-dimethoxybenzamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13h was prepared (yield: 43%). 1 H NMR (500MHz, CDCl3) δ12.05(s,1H),8.43(s,1H),8.29(s,1H),7.73(t,J=6.4H z,2H),7.60(d,J=8.1Hz,2H),7.47(dd,J=13.6,7.9Hz,1H),7.29(d,J=8.1Hz, 2H),7.25–7.21(m,1H),7.02(d,J=2.1Hz,2H),6.88(s,1H),3.92(s,3H),3.91 (s,3H),3.84(s,3H),3.76–3.72(m,5H),2.97–2.85(m,6H),2.77–2.71(m,2H).

[0222] 13 C NMR (101MHz, CDCl3) δ167.41,164.35,152.74,149.15,147.05,146.21,144 .55,135.58,135.50,130.58,130.50,129.48,122.72,122.69,121.29,119 .94,119.07,118.86,114.96,114.73,112.32,111.42,109.57,105.01,100.54,95.67,59.35,56.45,56.42,56.32,56.15,50.66,50.32,33.74,21.01.

[0223] Preparation Example 51

[0224] Preparation of N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxy-2-(pyridine-3-sulfonyl)benzamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13i was prepared (yield: 45%). 1 H NMR (500MHz, CDCl3) δ8.76(d,J=1.5Hz,1H),8.60(d,J=3.8Hz,1H),8.06(d,J=8.0Hz,1H),7.70(s,1H),7.33(d,J=7.8Hz,2H),7.30–7.17(m,5H) ,7.01(s,1H),6.91(s,1H),6.87(s,1H),3.93(s,3H),3.91(s,3H),3.90 (s,3H),3.84(s,3H),3.74(s,2H),2.96–2.85(m,6H),2.76–2.71(m,2H). 13 C NMR (101MHz, CDCl3) δ166.11,153.31,152.70,149.95,149.14,147.93,147.05,146.37,146.20,137.22,135.58,135.13,134.98,132.48,129 .39,123.77,121.06,119.92,115.95,111.42,109.23,107.62,100.53, 95.67,59.21,56.47,56.42,56.35,56.30,50.62,50.30,33.65,20.97.

[0225] Preparation Example 52

[0226] Preparation of N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoxaline-2-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13j was prepared (yield: 43%). 1 H NMR (400MHz, CDCl3+CD3OD) δ9.42(s,1H),8.32(s,1H),8.14–8.08(m,1H),7.94(d,J=8.1Hz,1H),7.73–7.63(m,2H),7.43(d,J=7.4Hz,2H),7.18(s,1H ),7.06(d,J=7.9Hz,2H),6.82(s,1H),6.70(s,1H),3.81(s,3H),3.75(s,3 H),3.70(s,3H),3.69(s,3H),3.57(s,2H),2.83–2.65(m,6H),2.57(s,2H). 13 C NMR (101MHz, CDCl3+CD3OD) δ171.45,165.96,155.85,153.26,153.12,151.13 ,150.16,148.69,148.08,147.23,144.53,140.35,139.83,137.44,136.07,1 35.01,134.37,132.97,132.61,125.68,123.73,118.61,115.27,115.10,108.88,104.81,99.76,63.07,60.29,60.09,59.85,54.38,53.95,36.99,24.35.

[0227] Preparation Example 53

[0228] Preparation of N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-2-oxoethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a pale yellow solid compound 13k was prepared (yield: 44%). 1H NMR (500 MHz, CDCl3, conformer) δ12.58–12.50 (m, 1H), 9.53–9.49 (m, 1H), 8.76–8.72 (m, 1H), 8.68–8.60 (m, 1H), 8.53–8.50 (m, 1H), 8.16–8.12 (m, 1H), 7.98–7.94 (m, 1H), 7.83–7.77 (m, 1H), 7.63–7.59 (m, 1H), 7.5 9–7.52(m,2H),7.23–7.16(m,3H),7.01–6.95(m,1H),6.87–6.79(m,1H),4.80–4.56(m,2H),3.96–3.92 (m,4H),3.91–3.88(m,3H),3.88–3.85(m,3H),3.83–3.80(m,4H),3.79–3.75(m,2H),2.71–2.56(m,2H). 13C NMR (126 MHz, CDCl3, conformer) δ 170.50, 170.44, 167.54, 167.52, 163.89, 163.85, 152.70, 152.66, 149.34, 149.31, 148.75, 148.66, 148.20, 147.62, 147.49, 146.81, 146.54, 146.42, 144.55, 136.50, 136.45, 135.97, 135.84, 135.73, 135.68, 131.47, 131.45, 131.15, 130.92, 129.3 6,129.33,129.26,129.21,129.18,127.48,127.46,127.29,127.26,126.95,126.92,121.52,119.30,119.23,113.06,112.16,112.10,111.32,110.01,104.93,100.59,100.44,95.74,95.63,56.43,56.39,56.29,56.13,44.23,44.03,40.93,40.85,40.55,39.94,21.67,20.60.

[0229] Preparation Example 54

[0230] Preparation of N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-difluorophenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 131 was prepared (yield: 39%).1 H NMR (400MHz, CDCl3) δ12.49(s,1H),9.50(s,1H),8.97–8.65(m,3H),8.16(d,J=6.9Hz,1H),7.99(d,J=6.6Hz,1H),7.90–7. 77(m,1H),7.74–7.54(m,4H),7.27(s,2H),7.01(s,1H),6.88(s,1H),3.92(s,6H),3.74(s,2H),2.92(bs,6H),2.73(s,2H). 13 C NMR (101MHz, CDCl3) δ166.05,163.94,150.04,149.53,149.15,148.59,147.06,146.22,137.48,136.16,135.22,131.70,129.42,129.23,127. 61,126.90,126.77,121.50,119.97,116.83,116.13,111.43,111.32,1 11.09,100.58,95.69,59.30,56.46,56.32,50.63,50.33,33.75,21.00.

[0231] Preparation Example 55

[0232] Preparation of N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)phenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13m was prepared (yield: 43%). 1H NMR (400MHz, CDCl3) δ12.08(s,1H),9.51(d,J=1.3Hz,1H),8.87(s,1H),8.76(s,1H),8.60(d,J=8.3Hz ,1H),8.18(d,J=8.4Hz,1H),8.00(d,J=8.1Hz,1H),7.83(t,J=7.5Hz,1H),7.70(d,J=8.1Hz,2H),7.64( t,J=7.5Hz,1H),7.59(d,J=7.6Hz,1H),7.37(t,J=7.8Hz,1H),7.29(d,J=8.1Hz,2H),7.00(s,1H),6.90 (t,J=7.6Hz,1H),6.87(s,1H),3.90(s,3H),3.89(s,3H),3.74(s,2H),2.99–2.85(m,6H),2.73(s,2H). 13 C NMR (101MHz, CDCl3) δ167.47,164.18,150.08,149.48,149.15,148.70,147 .04,146.20,139.08,137.01,136.05,135.77,132.59,131.62,129.47,129. 21,127.59,127.28,127.14,126.95,123.38,121.95,121.55,120.92,119.98,111.42,100.58,95.70,59.37,56.44,56.31,50.63,50.33,33.76,21.01.

[0233] Preparation Example 56

[0234] Preparation of N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxopropyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13n was prepared (yield: 46%).

[0235] 1H NMR (500 MHz, CDCl3, conformer) δ12.51 (s, 1H), 9.52 (s, 1H), 8.75 (s, 1H), 8.56 (s, 1H), 8.38–8.22 (m, 1H), 8.15 (d, J = 8.4 Hz, 1H), 7.98 (d, J = 8.1 Hz, 1H), 7.84–7.79 (m, 1H), 7.62 (t, J = 7.5 Hz, 1H), 7.55 (d, J = 8.3 Hz, 1H), 7.52 (d, J = 8.3 Hz, 1H ),7.24(t,J=8.2Hz,2H),7.15–7.07(m,1H),7.05–6.95(m,1H),6.89–6.80(m,1H),4.62(m,2H),3.95(s,3H),3.94–3 .91(m,1H),3.91–3.84(m,6H),3.81(s,3H),3.72(t,J=5.4Hz,1H),3.00(dd,J=15.8,8.0Hz,2H),2.79–2.65(m,4H). 13 C NMR (126 MHz, CDCl3, conformer) δ 171.47, 171.32, 167.51, 167.42, 163.85, 152.83, 149.35, 149.29, 148.79, 148.73, 148.53, 147.57, 147.44, 147.04, 146.50, 146.41, 144.61, 138.08, 137.95, 135.92, 135.85, 135.80, 135.57, 131.47, 129.35, 129.20, 129.09,127.49,127.30,126.95,121.46,121.32,119.37,119.31,113.09,112.08,111.29,109.79,105.00,100.56,100.42,95.75,95.58,56.45,56.41,56.28,56.25,56.16,43.68,43.35,40.78,39.71,35.78,35.33,30.80,30.78,21.74,20.60.

[0236] Preparation Example 57

[0237] Preparation of (E)-N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13o was prepared (yield: 38%). 1 H NMR (400 MHz, CDCl3 + CD3OD, conformer) δ9.40 (s, 1H), 8.70 (s, 1H), 8.43 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H), 7.94 (d, J = 8.1 Hz, 1H), 7.76 (t, J = 7.6 Hz, 1H), 7.68 (d, J = 8.3 Hz, 2H), 7.63–7.51 (m,2H),7.47(d,J=8.2Hz,2H),7.25–7.22(m,1H),7.01–6.93(m,1H),6.89–6.75(m,2H), 4.75(s,2H),3.90(s,3H),3.85(s,6H),3.82(s,3H),2.82(s,2H),2.73(d,J=21.9Hz,2H). 13 C NMR (101 MHz, CDCl3 + CD3OD, conformer) δ 167.82, 166.87, 163.60, 152.60, 149.33, 148.91, 148.51, 148.17, 147.51, 146.97, 146.40, 144.45, 142.92, 139.51, 136.09, 135.37, 131.72, 131.15, 129.22, 128.80,128.65,127.65,127.17,126.95,121.14,119.33,116.21,115.89,112.17,110.56,104.67,100.58,100.49,95.72,95.62,56.40,56.30,56.25,56.06,44.05,43.95,41.41,21.91,20.56.

[0238] Preparation Example 58

[0239] Preparation of (E)-N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamoyl)phenyl)quinoline-3-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13p was prepared (yield: 46%). 1H NMR (400 MHz, CDCl3+CD3OD, conformer) δ9.45 (s, 1H), 8.84 (s, 1H), 8.71 (d, J=8.3 Hz, 1H), 8.14 (d, J=8.4 Hz, 1H), 8.07 (d, J=8.1 Hz, 1H), 7.95–7.85 (m, 2H), 7.80 (d, J=8.3 Hz, 2H), 7.73–7.59 (m, 5H), 7.27 (t, J=7.6 Hz, 1H), 7.12–6.86 (m, 3H), 4.83 (s, 2H), 4.08–3.97 (m, 2H), 3.92 (s, 6H), 2.98–2.68 (m, 2H). 13 C NMR (101 MHz, CDCl3 + CD3OD, conformer) δ 165.64, 164.48, 161.12, 146.69, 145.95, 145.71, 145.34, 144.67, 144.32, 143.60, 140.39, 136.91, 136.33, 133.68, 130.18, 129.29, 128.60, 126.5 7,125.94,125.61,125.45,125.16,124.55,124.34,120.99,118.87,118.80,118.58,116.72,113.20,110.31,108.89,97.98,93.06,53.60,53.41,41.28,38.66,37.67,19.10,17.78.

[0240] Preparation Example 59

[0241] Preparation of N-(4-(4,5-dimethoxy-2-(quinoline-3-carboxamido)benzamido)phenyl)-6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridine-2(1H)-carboxamide: Referring to the preparation method in Preparation Example 33, a light yellow solid compound 13q was obtained (yield: 43%). 1H NMR (400MHz, CDCl3+CD3OD) δ9.35(s,1H),8.70(s,1H),8.42(s,1H),8.03(d,J=8.4Hz,1H) ,7.92(d,J=8.0Hz,1H),7.74(t,J=7.5Hz,1H),7.54(t,J=7.4Hz,1H),7.46(d,J=8.2Hz,2H ),7.29(s,1H),7.20(d,J=8.2Hz,2H),6.91(s,1H),6.79(s,1H),4.47(s,2H),4.31(s,2H) ,3.91(s,3H),3.84(s,3H),3.81(s,3H),3.80(s,3H),3.66(t,J=5.1Hz,2H),2.63(s,2H). 13 C NMR (101MHz, CDCl3+CD3OD) δ168.01,163.58,158.22,152.39,149.22,148.72, 148.46,148.12,147.26,146.24,144.48,136.56,136.13,135.24,131.74,129. 18,128.54,128.01,127.65,127.30,126.97,121.98,119.51,112.26,110.64,104.66,100.58,95.65,56.35,56.27,56.19,56.01,44.29,42.26,41.61,20.96.

[0242] In addition, it should be understood that after reading the above description of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound represented by formula (I) or (II) or a pharmaceutically acceptable salt thereof: R 1 、R 2 、R 3 、R 4 、R 6 、R 7 are the same or different and are independently selected from H, halogen, C1-C5 alkyl or C1-C5 alkoxy; R 5 、R 8 are the same or different and are independently selected from substituted or unsubstituted phenyl, pyridyl, pyrazinyl, quinolyl, isoquinolyl or quinoxalinyl; the substituents on the phenyl, pyridyl, pyrazinyl, quinolyl, isoquinolyl or quinoxalinyl are independently selected from cyano or halogen; m is selected from 0 to 4; X is selected from -NHC(=O)- or -NHS(=O)2-; L is selected from n are independently selected from 0 to 4; One end of the benzene ring in L is connected to an amide bond.

2. The compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound is selected from any one of the following: N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine; N-(4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine; N-(4-(2-(7,8-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine; N-(4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine; N-(4-(2-(3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-3-yl)quinazolin-4-amine; N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine; N-(4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine; N-(4-(2-(7,8-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine; N-(4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine; N-(4-(2-(3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(pyridin-4-yl)quinazolin-4-amine; N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; N-(2-((4-((6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; N-(2-((4-(2-(7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; N-(2-((4-((7,8-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)methyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; N-(2-((4-(2-(3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; 2-(3-cyanobenzamido)-N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxybenzamide; N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)nicotinamide; N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-2-(3-fluorobenzamido)-4,5-dimethoxybenzamide; N-(4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)-4,5-dimethoxy-2-(pyridine-3-sulfonyl)benzamide; N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-2-carboxamide; N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-2-oxoethyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)-4,5-difluorophenyl)quinoline-3-carboxamide; N-(2-((4-(2-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)ethyl)phenyl)carbamoyl)phenyl)quinoline-3-carboxamide; N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxopropyl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; (E)-N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofuran[2,3-c]pyridin-2(1H)-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamoyl)-4,5-dimethoxyphenyl)quinoline-3-carboxamide; (E)-N-(2-((4-(3-(6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridin-2(1H)-yl)-3-oxoprop-1-en-1-yl)phenyl)carbamoyl)phenyl)quinoline-3-carboxamide; N-(4-(4,5-dimethoxy-2-(quinoline-3-carboxamido)benzamido)phenyl)-6,7-dimethoxy-3,4-dihydrobenzofurano[2,3-c]pyridine-2(1H)-carboxamide.

3. A method for preparing the compound of formula (I) or (II) according to claim 1 or 2, characterized in that: Synthesis route: Compound 10 is a compound represented by formula (I), wherein Y is Compound 13 is a compound represented by formula (II), wherein Y is -NH-Z is -XR 8 .

4. The preparation method according to claim 3, characterized in that Compound 1 undergoes ortho-Friedel-Crafts acylation under acetic acid / boron trifluoride etherate conditions to obtain compound 2, compound 2 reacts with ethyl bromoacetate in the presence of cesium carbonate as a base to generate compound 3, compound 3 undergoes hydrolysis reaction to obtain compound 4, compound 4 undergoes cyclization reaction to obtain compound 5, compound 5 undergoes a multi-component reaction with benzylamine and formaldehyde aqueous solution in acetic acid solvent to obtain compound 6, compound 6 is debenzylated to obtain compound 7, compound 7 undergoes a nucleophilic substitution reaction with a halide or an amide condensation reaction to obtain compound 8, compound 8 undergoes a nitro reduction reaction to obtain compound 9; compound 9 undergoes a nucleophilic substitution reaction with compound 14 to obtain compound 10; alternatively, compound 9 undergoes an amide condensation reaction with compound 15 to obtain compound 11, compound 11 undergoes a nitro reduction reaction to obtain compound 12, and compound 12 undergoes an amide condensation reaction or a sulfonamide condensation reaction to obtain compound 13.

5. Use of the compound represented by formula (I) or (II) according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of a P-glycoprotein inhibitor.

6. The use according to claim 5, characterized in that The P-glycoprotein inhibitor is an oral preparation.

7. Use of the compound of formula (I) or (II) according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of a drug resistance regulator for improving the therapeutic effect of an anticancer drug.

8. The use according to claim 7, characterized in that The anticancer drugs are drugs that are not easily absorbed in the digestive tract due to the inhibitory effect of intestinal P-glycoprotein, including taxanes, vinca alkaloids, anthracyclines, camptothecin, podophyllotoxin, mitoxantrone, actinomycin, and colchicine; The taxanes include paclitaxel and docetaxel; Said vinca alkaloids include vincristine and vinblastine; The anthracyclines include daunorubicin and doxorubicin; The camptothecins include topotecan and irinotecan.

9. The use according to claim 7, characterized in that The cancer is a solid tumor or a hematological malignancy selected from leukemia, multiple myeloma, and lymphoma; The leukemia is acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, or chronic myeloid leukemia; The lymphoma is Hodgkin's lymphoma, non-Hodgkin's lymphoma, mantle cell lymphoma, follicular lymphoma, B cell lymphoma, T cell lymphoma, and diffuse large B cell lymphoma.

10. The use according to claim 7, characterized in that The drug resistance regulator is an oral preparation.

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