N, 4-diphenylpyrimidine-2-amine compound as well as preparation and application thereof

Abstract

The invention discloses an N, 4-diphenylpyrimidine-2-amine compound and preparation and application thereof.The N, 4-diphenylpyrimidine-2-amine compound can remarkably reverse tumor MDR, enhance the sensitivity of malignant tumor cells to anti-tumor drugs such as doxorubicin hydrochloride and mitoxantrone and improve the drug-resistant tumor apoptosis rate, has good potential in the aspect of MDR reversing and has important development prospects.

Description

(I) TECHNICAL FIELD

[0001] The present application belongs to the field of synthetic chemistry and pharmacotherapy, and particularly relates to a N,4-diphenylpyrimidin-2-amine compound, a preparation method and application thereof. (II) BACKGROUND

[0002] Multidrug resistance (MDR) is one of the key factors for the failure of chemotherapy of malignant tumors, which greatly limits the efficacy of anticancer drugs, leading to increased difficulty in treatment and poor prognosis of patients. The formation mechanism is complex, mainly involving enhanced drug efflux mechanism in cells, activated DNA repair ability, inhibited apoptosis pathway, and changes in tumor microenvironment, and other factors.

[0003] At present, the reversal agents for MDR mainly include: chemical synthesis of small molecule reversal agents (such as verapamil), natural products and their derivatives (such as cyclosporin A, flavonoids), and drug delivery systems based on nano-carriers. However, most of the existing MDR reversal agents still have obvious limitations, such as low pharmacological selectivity, insufficient in vivo inhibitory activity, strong self-toxicity, and possible interference with the metabolic kinetics of co-administered anticancer drugs. In addition, there is a compensation and cross-activation phenomenon between tumor drug resistance pathways, and the development of reversal agents targeting a single mechanism is easy to trigger bypass drug resistance activation, leading to non-persistent or ineffective reversal effect, which is still the main challenge of current MDR reversal therapy. (III) SUMMARY

[0004] The purpose of the present application is to provide a N,4-diphenylpyrimidin-2-amine compound, a preparation method and application thereof, which can significantly reverse MDR of tumors, enhance the sensitivity of malignant tumor cells to doxorubicin hydrochloride, mitoxantrone and other antitumor drugs, and improve the apoptosis rate of drug-resistant tumors, showing good potential in reversing MDR and having important development prospects.

[0005] The technical solution adopted by the present application is:

[0006] In a first aspect, the present application provides a N,4-diphenylpyrimidin-2-amine compound, the structure of which is shown in formula I:

[0007]

[0008] R is one or more substitutions, specifically methoxy, ethoxy, propoxy, butoxy, phenoxy, trifluoromethoxy, fluorine, chlorine, bromine, iodine, trifluoromethyl, methyl, ethyl, propyl, butyl, tert-butyl, cyclohexyl, phenyl, naphthyl, pyridyl, nitro, amino or cyano.

[0009] R is hydrogen, 2-methoxy, 3-methoxy, 4-methoxy, 2-ethoxy, 4-ethoxy, 4-propoxy, 4-phenoxy, 2,3-dimethoxy, 2,4-dimethoxy, 2,5-dimethoxy, 3,4-dimethoxy, 3,5-dimethoxy, 3,4,5-trimethoxy, 2-methyl, 3-methyl, 4-methyl, 3,5-dimethyl, 4-ethyl, 4-propyl, 4-tert-butyl, 4-cyclohexyl, 4-phenyl, 2-naphthyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-fluoro, 3-fluoro, 4-fluoro, 2-chloro, 3-chloro, 4-chloro, 2-bromo, 3-bromo, 4-bromo, 2-iodo, 3-iodo, 4-iodo, 2-nitro, 3-nitro, 4-nitro, 4-amino, 4-cyano, 2-trifluoromethyl, 3-trifluoromethyl, 4-trifluoromethyl, 3,5-ditrifluoromethyl, 2-trifluoromethoxy, 3-trifluoromethoxy, 4-trifluoromethoxy, 2-ethoxy, 3-ethoxy, or 3,4,5-trimethoxy, more preferably 4-fluoro.

[0010] In a second aspect, the present application also provides a method for preparing the N,4-diphenylpyrimidin-2-amine compound, which comprises the following steps:

[0011] S1, reacting compound II with p-aminobenzoic acid hydrochloride in the presence of a diluent, a base and a palladium catalyst at 60-160°C for 6-24 hours, and separating and purifying the reaction solution to obtain intermediate III; the diluent is an inert organic solvent; the base is an organic base or an inorganic base; and the catalyst is a zero-valent or divalent palladium catalyst;

[0012] S2, reacting intermediate III with 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride in the presence of a diluent, an acid-binding agent and a condensing agent at 0-50°C for 10-40 hours, and separating and purifying the reaction solution to obtain intermediate IV; the condensing agent is dicyclohexyl carbodiimide (DCC), diisopropyl carbodiimide (DIC), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide (EDCI), benzotriazole-1-yl oxytris(dimethylamino) phosphonium hexafluorophosphate (BOP), (benzotriazole-1-yl oxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-hydroxybenzotriazole (HOBt), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-benzotriazol-tetramethyluronium hexafluorophosphate (HBTU), O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), triphenyl phosphorus-polyhalogenated methane, triphenyl phosphorus-hexachloroacetone, triphenyl phosphorus-NBS, 3-acyl-2-thiazoline, etc.; the diluent is the same as in step S1; and the acid-binding agent is an organic base or an inorganic base;

[0013] S3, reacting compound V with an acetal agent at 60-160°C for 1-10 hours to obtain intermediate VI; the acetal agent is N,N-dimethylformamide dimethyl acetal (DMF-DMA), ethylene glycol dimethyl ether, triethyl orthoformate, formic anhydride, chloroformate, 1,1'-carbonyldiimidazole, or methanesulfonimidate; more preferably, the acetal agent is N,N-dimethylformamide dimethyl acetal (DMF-DMA);

[0014] S4, reacting compound VII with 50% aqueous cyanamide in the presence of a diluent and an acid at 70-140°C for 10-48 hours, and recrystallizing in a recrystallization reagent to obtain intermediate VIII; the diluent is the same as in step S1; the acid is an organic acid or an inorganic acid; the recrystallization reagent is an ether;

[0015] S5, reacting intermediate VI with intermediate VIII in the presence of a base and a dehydrating agent at 60-290°C for 10-60 hours to obtain intermediate IX; the base is the same as in step S1; the dehydrating agent is an alcohol compound;

[0016] S6, reacting intermediate IV with intermediate IX in the presence of a diluent, an acid-binding agent, and a condensing agent at 0-50°C for 10-40 hours to obtain compound I; the diluent is the same as in step S1; the acid-binding agent is the same as in step S2; the condensing agent is the same as in step S2;

[0017]

[0018] R in V, VI, IX, and X is the same as R in I.

[0019] All the intermediates and target compounds described above can be purified by conventional separation techniques such as recrystallization or chromatographic separation.

[0020] i.e., by undergoing the following reaction:

[0021]

[0022] Further, in steps S1-S7, the diluents can be the same or different, preferably the diluents are one of benzene, toluene, xylene, chlorobenzene, dichlorobenzene, petroleum ether, hexane, cyclohexane, dichloromethane, chloroform, carbon tetrachloride, diethyl ether, diisopropyl ether, dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, acetone, butanone, methyl isobutyl ketone, acetonitrile, propionitrile, butyronitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methyl-formanilide, N-methylpyrrolidone, hexamethylphosphoric triamide, methyl acetate, ethyl acetate, dimethyl sulfoxide, methanol, ethanol, 80-90% by volume aqueous ethanol solution, n-propanol, n-butanol, isopropanol, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, diethylene glycol monomethyl ether, or diethylene glycol monoethyl ether; more preferably the diluents are dichloromethane, n-butanol, or tetrahydrofuran.

[0023] Further, in step S1, the base is sodium hydroxide, potassium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, sodium methoxide, sodium ethoxide, trimethylamine, pyridine, N,N-dimethylaniline, N,N-dimethylbenzylamine, N-methylpiperidine, or N-methylmorpholine; and the catalyst is Pd(OAc)2, PdCl2, Pd(MeCN)2Cl2, Pd(PPh3)4, or Pd(TFA)2, etc.

[0024] Further, in step S1, the diluent is dichloromethane, the base is potassium carbonate, and the catalyst is tetrakis(triphenylphosphine)palladium, i.e., Pd(PPh3)4.

[0025] Further, in step S2, the acid scavenger is N,N-diisopropylethylamine (DIPEA), pyridine, N,N-dimethylaniline, N,N-dimethylbenzylamine, N-methylpiperidine, N-methylmorpholine, N,N-dimethylaminopyridine, diazabicyclooctane, diazabicyclononene, or diazabicycloundecene, sodium hydroxide, sodium ethoxide, trimethylamine, or triethylamine.

[0026] Further, in step S2, the diluent is dichloromethane, the acid scavenger is triethylamine, and the condensing agent is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI).

[0027] Further, in step S4, the acid is one or more of nitric acid, sulfuric acid, hydrochloric acid, phosphoric acid, p-toluenesulfonic acid, or trifluoroacetic acid; and the recrystallization reagent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether, isopropyl ether, or petroleum ether.

[0028] Further, in step S4, the diluent is n-butanol, the acid is 16N concentrated nitric acid, and the recrystallization reagent is methyl tert-butyl ether.

[0029] Further, in step S5, the dehydrating agent is methanol, ethanol, n-propanol, isopropanol, n-butanol, t-butanol, sec-butanol, n-pentanol, iso-pentanol, benzyl alcohol, phenethyl alcohol, cyclohexanol, trifluoroethanol, 2-methoxyethanol, ethylene glycol, glycerol or 1,2-propanediol.

[0030] Further, in step S5, the base is sodium hydroxide and the dehydrating agent is n-butanol.

[0031] Further, in step S6, the acid binding agent is triethylamine, the diluent is dichloromethane and the condensing agent is EDCI.

[0032] Further, in step S1, the reaction temperature can be in a wide range according to the selection of the diluent, preferably 80-100°C for 12h. The molar ratio of the compound II to the p-aminobenzoic acid hydrochloride is 1:1-1.5, preferably 1:1; the molar ratio of the compound II to the catalyst is 1:0.001-0.1, preferably 1:0.006; the molar ratio of the compound II to the base is 1:2-5, preferably 1:3; the volume of the diluent is 1-5mL / mmol of the compound II, preferably 3mL / mmol.

[0033] Further, in step S1, the following procedure is used: under nitrogen protection, the p-aminobenzoic acid hydrochloride, the catalyst, the base and the compound II are added into a three-necked flask with a thermometer, and the mixture is reacted at 80°C for 12h; the reaction solution is filtered through a qualitative filter paper with a pore size of 50 microns, and the filtrate is collected, dried with sodium sulfate overnight after water washing, and then the drying agent is filtered out and the solvent is removed by a rotary evaporator under reduced pressure to obtain a solid; the solid is dissolved in dichloromethane and then loaded into a silica gel chromatography column (silica gel 200-300 mesh, column height 10cm, diameter 2cm), eluted with petroleum ether: ethyl acetate (10:1 by volume) at a rate of 1 / 10 column volume per minute, and monitored by thin layer chromatography with petroleum ether: ethyl acetate (2:1 by volume) as the developing agent; the eluate with an Rf of 0.5 is collected, and the solvent is removed by a rotary evaporator under reduced pressure to obtain the intermediate III; the base is potassium carbonate; the catalyst is tetrakis(triphenylphosphine)palladium; the diluent is dichloromethane; the molar ratio of the compound II to the p-aminobenzoic acid hydrochloride is 1:1; the molar ratio of the compound II to the catalyst is 1:0.006; the molar ratio of the compound II to the base is 1:3; the volume of the diluent is 3mL / mmol of the compound II.

[0034] Further, the reaction temperature of step S2 can be in a wide range according to the selection of the diluent, preferably 0-30°C for 24h. The molar ratio of intermediate III to 6,7-dimethoxy-l,2,3,4-tetrahydroisoquinoline hydrochloride is 1:1-1.5, preferably 1:1; the volume of the diluent is 1-5mL / mmol based on the amount of substance of intermediate III, preferably 3mL / mmol; the molar ratio of intermediate III to acid-binding agent is 1:1-3, preferably 1:2; the molar ratio of intermediate III to condensing agent is 1:0.5-2, preferably 1:1.

[0035] Further, step S2 is carried out as follows: in a three-necked flask equipped with a thermometer, 6,7-dimethoxy-l,2,3,4-tetrahydroisoquinoline hydrochloride and a diluent are added, stirred and cooled to 0°C, acid-binding agent is added dropwise, the dropping speed is controlled to keep the temperature at 0°C; after the addition is completed, the system is stirred for 30min, the temperature is maintained at 0°C, condensing agent is added, and stirred for 30min; the temperature is maintained at 0°C, intermediate III is added, and then slowly raised to room temperature, reacted at 25°C for 18-24h, the reaction solution is transferred to a separatory funnel, extracted with an equal volume of saturated brine for 3 times, the organic phase is combined, dried with anhydrous sodium sulfate overnight, filtered to remove the drying agent with qualitative filter paper with a pore size of 50 microns, the filtrate is collected, and the solvent is removed by rotary evaporator under reduced pressure to obtain a light yellow solid; the light yellow solid is dissolved in dichloromethane, and then loaded on a silica gel chromatography column (silica gel 200-300 mesh, column height 10cm, diameter 2cm), eluted with petroleum ether: ethyl acetate (5:1 by volume) as eluent, the elution speed is 1 / 10 column volume per minute, eluted for 3 column volumes, and monitored by thin layer chromatography with petroleum ether: ethyl acetate (1:1 by volume) as developing agent, the effluent with Rf of 0.3 is collected, and the solvent is removed by rotary evaporator under reduced pressure to obtain intermediate IV; the acid-binding agent is triethylamine; the diluent is dichloromethane; the condensing agent is l-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI); the molar ratio of intermediate III to 6,7-dimethoxy-l,2,3,4-tetrahydroisoquinoline hydrochloride is 1:1, the volume of the diluent is 3mL / mmol based on the amount of substance of intermediate III, and the molar ratio of intermediate III to acid-binding agent is 1:2; the molar ratio of intermediate III to condensing agent is 1:1.

[0036] Further, in step S3, the reaction temperature can be in a wide range according to the selection of the acetal agent, preferably 90-120°C for 5h. The molar ratio of compound V to acetal agent is 1:1-10, preferably 1:1.5.

[0037] Further, step S3 is performed as follows: compound V and acetal agent are sequentially added into a single-mouth flask equipped with a condenser, the reaction system is placed in an oil bath heating device equipped with methyl silicone oil, the reaction temperature is monitored in real time by a thermocouple probe temperature sensor, and the reaction is refluxed at 103°C for 5 h; the temperature is reduced to room temperature, and the solvent is removed by rotary evaporation under reduced pressure to obtain a solid; the solid is dissolved in dichloromethane and then loaded onto a silica gel chromatographic column, petroleum ether: ethyl acetate (10:1 by volume) is used as an eluent, the elution rate is 1 / 10 column volume per minute, 3 column volumes are eluted, petroleum ether: ethyl acetate (1:1 by volume) is used as a developing agent for thin layer chromatography monitoring, the effluent with an Rf of 0.5 is collected, and the solvent is removed by rotary evaporation under reduced pressure to obtain intermediate VI; the acetal agent is N,N-dimethylformamide dimethyl acetal (DMF-DMA); and the mass ratio of the amount of compound V to the amount of acetal agent is 1:1.5.

[0038] Further, in step S4, the reaction temperature can be performed in a wide range according to the selection of the diluent, and is preferably 100-130°C for 16 h. The volume of the diluent is 3-10 mL / mmol of the amount of substance of compound VII, preferably 5 mL / mmol; the volume of the 50% aqueous cyanamide solution is 0.1-1 mL / mmol of the amount of substance of compound VII, preferably 0.3 mL / mmol; the mass ratio of the amount of compound VII to the amount of acid is 1:1-3, preferably 1:1.5; the volume of the recrystallization reagent for crystallization is 5-20 mL / mmol of the amount of substance of compound VII, preferably 10 mL / mmol; and the volume of the recrystallization reagent for elution is 0.5-2 mL / mmol of the amount of substance of compound VII, preferably 0.6 mL / mmol.

[0039] Preferably, step S4 is performed as follows: In a single-necked flask equipped with a condenser, compound VII and a diluent are added sequentially and stirred to dissolve. A 50% cyanamide aqueous solution and an acid are added dropwise sequentially. The reaction system is placed in an oil bath heating device equipped with a methyl silicone oil bath. The reaction temperature is monitored in real time using a thermocouple probe temperature sensor. The reaction is refluxed at 118°C for 16 hours. After cooling to room temperature, the reaction solution is poured into a recrystallization reagent pre-cooled to 0°C and stirred at 0°C for 30 minutes to induce crystallization. The system is filtered using qualitative filter paper with a pore size of 50 micrometers, and the filter residue is collected and filtered using a recrystallization reagent pre-cooled to 0°C. The reagent was used for rinsing, and the solvent was removed by vacuum distillation in a rotary evaporator to obtain intermediate VIII; the diluent was n-butanol; the acid was 16N concentrated nitric acid; the recrystallization reagent was methyl tert-butyl ether; the volume of the diluent used was 5 mL / mmol based on the molar amount of compound VII; the volume of the 50% cyanamide aqueous solution used was 0.3 mL / mmol based on the molar amount of compound VII; the molar ratio of compound VII to acid was 1:1.5; and the volume of the recrystallization reagent used for crystallization was 10 mL / mmol based on the molar amount of compound VII.

[0040] Furthermore, in step S5, the reaction temperature can be within a wide range depending on the selection of the dehydrating agent, preferably 70–120℃ for 48 hours. The molar ratio of intermediate VI to intermediate VIII is 1:1-3, preferably 1:1.3; the volumetric amount of dehydrating agent, based on the amount of intermediate VI, is 3-10 mL / mmol, preferably 4 mL / mmol; the molar ratio of intermediate VI to alkali is 1:2-5, preferably 1:3.

[0041] Further, step S5 is performed as follows: In a single-necked flask equipped with a condenser, intermediate VI, intermediate VIII, dehydrating agent, and alkali are added sequentially. The reaction system is placed in an oil bath heating device containing methyl silicone oil. The reaction temperature is monitored in real time by a thermocouple probe temperature sensor. The reaction is refluxed at 118°C for 48 hours. After cooling to room temperature, the reaction solution is transferred to a separatory funnel and extracted three times with an equal volume of saturated brine. The organic phases are combined and dried overnight with anhydrous sodium sulfate. The desiccant is removed by filtering with qualitative filter paper with a pore size of 50 micrometers. The filtrate is collected and the solvent is removed by vacuum distillation using a rotary evaporator to obtain a solid. The solid is dissolved in dichloromethane. After elution, the sample was loaded onto a silica gel column using pure dichloromethane as the eluent at a rate of 1 / 10 column volume per minute for elution of 3 column volumes. Thin-layer chromatography was then performed using a dichloromethane:methanol mixture (volume ratio 100:1) as the developing solvent. The eluent with an Rf of 0.5 was collected and the solvent was removed by vacuum distillation using a rotary evaporator to obtain intermediate IX. The alkali was sodium hydroxide; the dehydrating agent was n-butanol; the molar ratio of intermediate VI to intermediate VIII was 1:1.3; the volumetric amount of dehydrating agent was 4 mL / mmol based on the molar amount of intermediate VI; and the molar ratio of intermediate VI to alkali was 1:3.

[0042] Further, in step S6, the reaction temperature can be in a wide range according to the selection of the diluent, preferably 0-30℃ for 24h. The molar ratio of intermediate IV to intermediate IX is 1:1-1.5, preferably 1:1; the volume of the diluent is 1-5mL / mmol of intermediate IV, preferably 3mL / mmol; the molar ratio of intermediate IV to acid-binding agent is 1:1-3, preferably 1:2; the molar ratio of intermediate IV to condensing agent is 1:0.5-2, preferably 1:1.

[0043] Further, step S6 is carried out as follows: in a three-necked flask equipped with a thermometer, intermediate IV and diluent are added, stirred and cooled to 0℃, acid-binding agent is added dropwise, the dropping speed is controlled to keep the temperature at 0℃; after the dropwise addition is completed, stirring is continued for 30min, the temperature of the system is maintained at 0℃, condensing agent is added, and stirring is continued for 30min; the temperature of the system is maintained at 0℃, intermediate IX is added, and the temperature is slowly raised to room temperature, and the reaction is carried out at 25℃ for 18-24h; the reaction solution is transferred to a separatory funnel, extracted with an equal volume of saturated brine three times, the organic phases are combined, dried with anhydrous sodium sulfate overnight, the drying agent is filtered off with qualitative filter paper with a pore size of 50 microns, the filtrate is collected, the solvent is removed by rotary evaporator under reduced pressure, and a solid is obtained; the solid is dissolved in dichloromethane, and then loaded onto a silica gel chromatography column, eluted with petroleum ether: ethyl acetate (5:1 by volume) as eluent, at a rate of 1 / 10 column volume per minute, for 3 column volumes, and monitored by thin layer chromatography with petroleum ether: ethyl acetate (1:1 by volume) as developing agent, the effluent with an Rf of 0.3 is collected, and the solvent is removed by rotary evaporator under reduced pressure to obtain the compound of formula I; the acid-binding agent is triethylamine; the diluent is dichloromethane; the condensing agent is 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide; the molar ratio of intermediate IV to intermediate IX is 1:1; the volume of the diluent is 3mL / mmol of intermediate IV; the molar ratio of intermediate IV to acid-binding agent is 1:2; the molar ratio of intermediate IV to condensing agent is 1:1.

[0044] In a third aspect, the present application also provides a use of the N,4-diphenylpyrimidin-2-amine compound in the preparation of a tumor MDR reversing agent.

[0045] Further, the inhibitor is a tumor multi-drug resistance reversing agent and a tumor metastasis inhibitor. The tumor includes human breast cancer adriamycin-resistant cell line MCF-7 / ADR and human colon cancer mitoxantrone-resistant cell line HCT116 / ABCG2. The inhibitor also includes a pharmaceutically acceptable salt.

[0046] The compound of the present application can be combined with anti-tumor drugs (clinical chemotherapy drugs) such as alkylating agents (such as cyclophosphamide or cisplatin), antimetabolites (such as 5-fluorouracil or hydroxyurea), topoisomerase inhibitors (such as mitoxantrone, camptothecin or topotecan), cell microtubule inhibitors (such as paclitaxel or vinblastine), DNA inserters (such as doxorubicin or daunorubicin) and lysine kinase inhibitors (such as gefitinib or imatinib). By combined treatment with anti-tumor drugs, the sensitivity of multi-drug resistant tumor cells to anti-tumor drugs is enhanced, thereby improving the effect of chemotherapy treatment.

[0047] Compared with the prior art, the present application has the following beneficial effects:

[0048] (1) The present application provides a kind of N, 4-diphenyl pyrimidine-2-amine compound, preparation method is simple, total yield reaches 31%~54%, show good synthesis efficiency.

[0049] (2) The N, 4-diphenyl pyrimidine-2-amine compound prepared by the present application has good biological activity, shows obvious sensitization effect in low cytotoxicity concentration to cancer cell strains showing drug resistance, can effectively improve the drug efficacy of doxorubicin hydrochloride on human breast cancer doxorubicin-resistant cell strain (MCF-7 / ADR) and effectively improve the drug efficacy of tetrahydroisoquinoline-indole-pyrimidine on human colon cancer mitoxantrone-resistant cell strain HCT116 / ABCG2, the RF value range of MCF-7 / ADR cell strain combined with doxorubicin is 6.3~134.9, the RF value range of HCT116 / ABCG2 cell strain combined with mitoxantrone is 2.6~2664.2, can be used as tumor multi-drug resistance reversing agent and tumor metastasis inhibitor or biological probe with good biological inclusion, expands the application prospect of N, 4-diphenyl pyrimidine-2-amine structure in tumor multi-drug resistance reversing agent. Among them, compound I-22 shows better activity than the control drug cyclosporin A (Cyclosporin A) in the combined drug system, the multi-drug resistance reversing coefficient (RF) of doxorubicin combined with doxorubicin is as high as 134.9, and shows excellent anti-proliferation effect on MCF-7 / ADR cell strain; the activity of compound I-21 significantly exceeds that of the control drugs cyclosporin A and gefitinib, the RF value of doxorubicin combined with doxorubicin is 32.9, and the RF value of mitoxantrone combined with mitoxantrone is 2664.2, which shows excellent dual anti-proliferation effect on MCF-7 / ADR resistant cell strain and HCT116 / ABCG2 resistant cell strain, and has outstanding clinical application potential. (IV) BRIEF DESCRIPTION OF DRAWINGS

[0050] Figure 1 It is the nuclear magnetic resonance hydrogen spectrum of the compound shown in formula I-1. (V) DETAILED DESCRIPTION

[0051] The application will be further described in conjunction with specific examples. However, the scope of the application is not limited to the following examples:

[0052] Unless otherwise specified, the reagents and methods involved in the examples are the reagents and methods commonly used in the art.

[0053] The room temperature described in the application refers to 25-30°C.

[0054] Example 1, Preparation of Compound I

[0055] 1. Preparation of Compound I-1

[0056]

[0057] S1, under nitrogen protection, 5 mmol of p-aminobenzoic acid hydrochloride, 0.03 mmol of tetrakis(triphenylphosphine)palladium, 15 mmol of potassium carbonate and 5 mmol of compound II were sequentially added into a 50 mL three-necked flask equipped with a thermometer, and 15 mL of dichloromethane was added, and the reaction was carried out at 80°C for 12 h. The reaction solution was filtered with qualitative filter paper with a pore size of 50 microns, the filtrate was collected, washed with water and dried with sodium sulfate overnight, the drying agent was filtered out, and the solvent was removed by rotary evaporator under reduced pressure to obtain white solid. All the white solid was dissolved in 0.1 mL of dichloromethane and then loaded onto a silica gel chromatography column (silica gel 200-300 mesh, column height 10 cm, diameter 2 cm), eluted with petroleum ether: ethyl acetate (volume ratio 10:1) as eluent, elution rate was 1 / 10 column volume per minute, elution amount was 3 column volumes, thin layer chromatography was monitored with petroleum ether: ethyl acetate (volume ratio 2:1) as developing agent, the effluent with Rf of 0.5 was collected, and the solvent was removed by rotary evaporator under reduced pressure to obtain 4.9 mmol of intermediate III, i.e. 5-(4-aminophenyl)-1H-indole-2-carboxylic acid, which was used for subsequent reaction.

[0058] S2, in a 50 mL three-necked flask equipped with a thermometer, 6,7-dimethoxy- 1,2,3,4-tetrahydroisoquinoline hydrochloride (7.5 mmol) and 22.5 mL of dichloromethane were added, and the mixture was stirred to cool to 0°C, 15 mmol of triethylamine was added dropwise, and the dropping speed was controlled to keep the temperature at 0°C. After the addition was completed, the mixture was stirred for 30 min, and the temperature of the system was maintained at 0°C, 7.5 mmol of EDCI was added, and the mixture was stirred for 30 min; the temperature of the system was maintained at 0°C, 7.5 mmol of 5-(4-aminophenyl)-1H-indole-2-carboxylic acid (III) was added, and the mixture was slowly warmed to 25°C after the addition was completed, and the mixture was reacted at 25°C for 20 h. The reaction solution was transferred to a separatory funnel, and the mixture was extracted with an equal volume of saturated brine three times, and the organic phase was combined, dried over anhydrous sodium sulfate overnight, filtered to remove the drying agent using a qualitative filter paper with a pore size of 50 microns, the filtrate was collected, and the solvent was removed by rotary evaporation under reduced pressure to obtain a light yellow solid. The entire light yellow solid was dissolved in 0.1 mL of dichloromethane and then loaded onto a silica gel chromatography column (silica gel 200-300 mesh, column height 10 cm, diameter 2 cm), eluted with petroleum ether: ethyl acetate (5:1 by volume) as the eluent, at a flow rate of 1 / 10 column volume per minute, and eluted for 3 column volumes, and monitored by thin layer chromatography using petroleum ether: ethyl acetate (1:1 by volume) as the developing agent, the effluent with an Rf of 0.3 was collected, and the solvent was removed by rotary evaporation under reduced pressure to obtain 7.4 mmol of intermediate IV, i.e., (5-bromo-1H-indol-2-yl)(6,7-dimethoxy-3,4-dihydroisoquinolin-2(1H)-yl)methanone, which was used in the next step.

[0059] S3, in a 50 mL single-necked flask equipped with a condenser, 7.5 mmol of acetone (V-1), 10 mL of N,N-dimethylformamide dimethyl acetal (DMF-DMA) were added in sequence, the reaction system was placed in an oil bath heating device filled with methyl silicone oil, and the reaction temperature was monitored in real time by a thermocouple probe temperature sensor, and the reaction was carried out at 103°C for 5 h; the temperature was lowered to room temperature, and the solvent was removed by rotary evaporation under reduced pressure to obtain a light yellow oily liquid; the entire light yellow oily liquid was dissolved in 0.1 mL of dichloromethane and then loaded onto a silica gel chromatography column (silica gel 200-300 mesh, column height 10 cm, diameter 2 cm), eluted with petroleum ether: ethyl acetate (10:1 by volume) as the eluent, at a flow rate of 1 / 10 column volume per minute, and eluted for 3 column volumes, and monitored by thin layer chromatography using petroleum ether: ethyl acetate (1:1 by volume) as the developing agent, the effluent with an Rf of 0.5 was collected, and the solvent was removed by rotary evaporation under reduced pressure to obtain 7.4 mmol of intermediate VI-1, i.e., 4-(dimethylamino)but-3-en-2-one, which was used in the subsequent reaction.

[0060] S4, in a 100 mL single neck flask equipped with a condenser, 7.5 mmol of methyl 3-amino-4-methylbenzoate (VII) was added, followed by 37.5 mL of n-butanol and stirred to dissolve, 2.5 mL of 50% aqueous cyanamide solution and 11.5 mL of 16 N concentrated nitric acid were added dropwise, the reaction system was placed in an oil bath heating device filled with methyl silicone oil, the reaction temperature was monitored in real time by a thermocouple probe temperature sensor, and the reaction was refluxed at 118°C for 16 h; cooled to room temperature, the reaction liquid was poured into 75 mL of pre-cooled to 0°C recrystallization reagent methyl tert-butyl ether, stirred at 0°C for 30 min to induce crystallization, filtered with a qualitative filter paper with a pore size of 50 microns, washed with 5 mL of pre-cooled to 0°C methyl tert-butyl ether, and the solvent was removed by rotary evaporator under reduced pressure to obtain 7.4 mmol of intermediate VIII, i.e. methyl 3-guanidino-4-methylbenzoate, which was used for the next step reaction.

[0061] S5, in a 50 mL single neck flask equipped with a condenser, 5 mmol of intermediate VI-1, 6.5 mmol of intermediate VIII, 20 mL of n-butanol, and 15 mmol of sodium hydroxide were added, the reaction system was placed in an oil bath heating device filled with methyl silicone oil, the reaction temperature was monitored in real time by a thermocouple probe temperature sensor, and the reaction was refluxed at 118°C for 48 h, cooled to room temperature, the reaction liquid was transferred to a separatory funnel, extracted with an equal volume of saturated brine 3 times, the organic phase was combined, dried with anhydrous sodium sulfate overnight, filtered with a qualitative filter paper with a pore size of 50 microns to remove the drying agent, the filtrate was collected, and the solvent was removed by rotary evaporator under reduced pressure to obtain a white solid; all the white solid was dissolved in 0.1 mL of dichloromethane and then loaded onto a silica gel chromatography column (silica gel 200-300 mesh, column height 10 cm, diameter 2 cm), eluted with pure dichloromethane as eluent, elution rate was 1 / 10 column volume per minute, eluted for 3 column volumes, monitored by thin layer chromatography with dichloromethane:methanol at a volume ratio of 100:1 as developing agent, the effluent with Rf of 0.5 was collected, and the solvent was removed by rotary evaporator under reduced pressure to obtain 3.6 mmol of intermediate IX-1, i.e. 4-methyl-3-((4-methylpyrimidin-2-yl)amino)benzoic acid ethyl ester, which was used for the next step reaction.

[0062] S6. In a 25 mL three-necked flask equipped with a thermometer, add 2 mmol of intermediate IV and 6 mL of dichloromethane, stir and cool to 0 °C, then add 4 mmol of triethylamine dropwise, controlling the dropping rate to maintain the temperature at 0 °C; after the addition is complete, stir for 30 min, maintaining the system temperature at 0 °C, add 2 mmol of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), stir for 30 min; maintaining the system temperature at 0 °C, add 2 mmol of intermediate IX-1, slowly raise to room temperature, and react at 25 °C for 24 h. Transfer the reaction solution to a separatory funnel, extract three times with an equal volume of saturated saline, combine the organic phases, dry overnight with anhydrous sodium sulfate, filter to remove the desiccant using 50 μm qualitative filter paper, collect the filtrate, and remove the solvent by vacuum distillation using a rotary evaporator to obtain a solid; dissolve the solid in dichloromethane, and load it onto a silica gel chromatography column (200-300 mesh silica gel, 10 cm high, 2 cm diameter) to obtain a solid. Using a petroleum ether:ethyl acetate ratio of 5:1 (v / v) as the eluent, the elution rate was 1 / 10 column volume per minute. After eluting for 3 column volumes, thin-layer chromatography was performed using a petroleum ether:ethyl acetate ratio of 1:1 (v / v) as the developing solvent. The eluent with an Rf of 0.3 was collected, and the solvent was removed by rotary evaporation under reduced pressure to obtain 1.4 mmol of compound I-1, namely N-(4-(2-(6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)-1H-indol-5-yl)phenyl)-4-methyl-3-((4-methylpyrimidin-2-yl)amino)benzamide. The 1H NMR spectrum is shown below. Figure 1 The total molar yield of the seven-step reaction was 35%.

[0063] 2. Preparation of compounds I-2 to I-38

[0064] Following a similar method to step 1, only the R in compound V was replaced as shown in Table 1 to obtain the corresponding compounds I-2 to I-38. The appearance, yield, and high-resolution mass spectrometry data of these compounds are listed in Table 1, and the 1H NMR results are listed in Table 2. It can be seen from the above that the structures of the above compounds are correct, and they are all compounds I.

[0065] Table 1. Physicochemical constants and high-resolution mass spectrometry data of compound I.

[0066]

[0067]

[0068] Table 2. 1H NMR spectral data of compound I

[0069]

[0070]

[0071]

[0072]

[0073] Example 2 Study on Multidrug Resistance Reversal Activity of N,4-Diphenylpyrimidin-2-amine Compounds on MCF-7 / ADR Cells and HCT116 / ABCG2

[0074] Cell strain: MCF-7 / ADR (human breast cancer cell line resistant to adriamycin), HCT116 / ABCG2 (human colon cancer cell line resistant to mitoxantrone).

[0075] Test compound: compounds I-1 to I-38 prepared in Example 1.

[0076] Positive control drug: Cyclosporin A, Gefitinib.

[0077] Test method: MTT (tetrazolium salt) method was used to perform cell proliferation activity test of the compound combined with adriamycin (or mitoxantrone).

[0078] MCF-7 / ADR cells were cultured in RPMI 1640 medium containing 10% bovine serum at 37℃, 5% CO2 saturated humidity. Cells in logarithmic growth phase were inoculated in 96-well culture plates at a density of 1×10 5 / mL, 100 μL per well, and cultured at 37℃, 5% CO2 saturated humidity, and divided into blank control group, test compound group, and positive control group. Test compound, positive control drug, and adriamycin were respectively prepared into a solution with a concentration of 10 mM with dimethyl sulfoxide (DMSO) for standby. Different test compound solutions and adriamycin solutions were added to the test compound group, diluted with RPMI 1640 medium containing 10% bovine serum, and the final concentration of the test compound and adriamycin was 5 μmol / L; positive control drug solution and adriamycin solution were added to the positive control drug group, diluted with RPMI 1640 medium containing 10% bovine serum, and the final concentration of the positive control drug and adriamycin was 5 μmol / L; the blank control group was given the same volume of PBS. After 48 hours of culture, MTT working solution was added, and after 4 hours, the culture solution was poured out, 100 μL of DMSO was added to each well for dissolution, and then the optical density was read at a wavelength of 570 nm on an enzyme marker, and the IC 50 (μM) and the reversal fold (RF) of multidrug resistance were calculated, and the effect of the compound on the survival rate of the cells was investigated, and the results are shown in Table 3.

[0079] HCT116 / ABCG2 cells were cultured in RPMI 1640 medium containing 10% calf serum at 37°C, 5% CO2 saturated humidity. The cells in logarithmic growth phase were inoculated in 96-well culture plates at a density of 1 x 10 5 / mL, 100 μL per well, and cultured at 37°C, 5% CO2 saturated humidity. The cells were divided into blank control group, test compound group and positive control group. The test compound, positive control drug and mitoxantrone were respectively dissolved in dimethyl sulfoxide (DMSO) to prepare a solution with a concentration of 10 mM for standby. Different test compound solutions and mitoxantrone solutions were added to the test compound group, which were diluted with RPMI 1640 medium containing 10% calf serum, and the final concentrations of the test compound and mitoxantrone were both 5 μmol / L. The positive control drug solution and mitoxantrone solution were added to the positive control group, which were diluted with RPMI 1640 medium containing 10% calf serum, and the final concentrations of the positive control drug and mitoxantrone were both 5 μmol / L. The blank control group was given the same volume of PBS. After 48 hours of further culture, MTT working solution was added, and after 4 hours, the culture solution was poured out, 100 μL of DMSO was added to each well for dissolution, and then the optical density was read at a wavelength of 570 nm on an enzyme marker. The IC 50 (μM) and the reversal factor (RF) of multidrug resistance were calculated to investigate the effect of the compound on the survival rate of the cells, and the results are shown in Table 4.

[0080] Table 3 Results of the reversal activity of 5 μM of the compound of Formula I on MCF-7 / ADR cells

[0081]

[0082] a IC 50 value of doxorubicin combined with the test compound (5 μM). b Reversal factor (reversal fold) = IC 50 (ADR) / IC 50 (MDR inhibitor + ADR). c IC 50 (ADR) of doxorubicin alone. d N.T.: Not tested due to excessive toxicity (more than 30% inhibition) when used alone.

[0083] Table 4 Results of the reversal activity of 5 μM of the compound of Formula I on HCT116 / ABCG2 cells

[0084]

[0085]

[0086] IC of mitoxantrone combined with test compound (5 μM) 50 Reversal fold = IC 50 (MX) / IC 50 (MDR inhibitor + MX). c IC of mitoxantrone alone 50 (MX). d N.T.: Not tested due to toxicity (more than 30% inhibition) when used alone.

[0087] As can be seen from Tables 3 and 4, the tested compounds generally have good reversal of multi-drug resistance activity of tumor cells, among which, compound I-22 shows better activity than the control drug Cyclosporin A in the combined drug system, and has a multi-drug resistance reversal factor (RF) of 134.9 when combined with doxorubicin, and shows excellent anti-proliferation effect on MCF-7 / ADR cell strain; the activity of compound I-21 significantly exceeds that of the control drugs Cyclosporin A and Gefitinib, and has an RF value of 32.9 when combined with doxorubicin, and an RF value of 2664.2 when combined with mitoxantrone, and shows excellent dual anti-proliferation effect on MCF-7 / ADR and HCT116 / ABCG2 drug-resistant cell strains, and has outstanding clinical application potential. The rest of the compounds also show excellent drug resistance reversal effect, and have an RF value of 6.3-134.9 on MCF-7 / ADR cell strain when combined with doxorubicin, and an RF value of 2.6-2664.2 on HCT116 / ABCG2 cell strain when combined with mitoxantrone, and have good application prospect.

Claims

1. An N,4-diphenylpyrimidine-2-amine compound, the structure of which is shown in Figure I: In part I, the R group can be one or more substitutions, wherein the R group is methoxy, ethoxy, propoxy, butoxy, phenoxy, trifluoromethoxy, fluorine, chlorine, bromine, iodine, trifluoromethyl, methyl, ethyl, propyl, butyl, tert-butyl, cyclohexyl, phenyl, naphthyl, pyridyl, nitro, amino, or cyano.

2. The N,4-diphenylpyrimidine-2-amine compound as described in claim 1, characterized in that, The R group is a hydrogen group, 2-methoxy, 3-methoxy, 4-methoxy, 2-ethoxy, 4-ethoxy, 4-propoxy, 4-phenoxy, 2,3-dimethoxy, 2,4-dimethoxy, 2,5-dimethoxy, 3,4-dimethoxy, 3,5-dimethoxy, 3,4,5-trimethoxy, 2-methyl, 3-methyl, 4-methyl, 3,5-dimethyl, 4-ethyl, 4-propyl, 4-tert-butyl, 4-cyclohexyl, 4- Phenyl, 2-naphthyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-fluoro, 3-fluoro, 4-fluoro, 2-chloro, 3-chloro, 4-chloro, 2-bromo, 3-bromo, 4-bromo, 2-iodine, 3-iodine, 4-iodine, 2-nitro, 3-nitro, 4-nitro, 4-amino, 4-cyano, 2-trifluoromethyl, 3-trifluoromethyl, 4-trifluoromethyl, 3,5-ditrifluoromethyl, 2-trifluoromethoxy, 3-trifluoromethoxy, or 4-trifluoromethoxy.

3. A method for preparing the N,4-diphenylpyrimidine-2-amine compound according to claim 1 or 2, characterized in that, The method includes the following steps: S1. In the presence of a diluent, a base, and a palladium catalyst, compound II reacts with p-aminophenylborate acid salt at 60–160°C for 6–24 hours, and the reaction solution is separated and purified to obtain intermediate III; the diluent is an inert organic solvent; the base is an organic or inorganic base; and the catalyst is a zero-valent or divalent palladium catalyst. S2. In the presence of a diluent, an acid-binding agent, and a condensing agent, intermediate III reacts with 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride at 0–50 °C for 10–40 hours. The reaction solution is then separated and purified to obtain intermediate IV. The condensing agent is dicyclohexylcarbodiimide, diisopropylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, benzotriazole-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate, or (benzotriazole-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate. The following substances are used: pyrrolidinylphosphonium hexafluorophosphate, 1-hydroxybenzotriazole, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, O-benzotriazole-tetramethylurea hexafluorophosphate, O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, triphenylphosphine-polyhalomethane, triphenylphosphine-hexachloroacetone, triphenylphosphine-NBS, or 3-acyl-2-thiothiazoline; the diluent is the same as in step S1; the acid-binding agent is an organic or inorganic base. S3. In the presence of an acetal agent, compound V is reacted at 60–160°C for 1–10 hours, and the reaction solution is separated and purified to obtain intermediate VI; the acetal agent is N,N-dimethylformamide dimethyl acetal, ethylene glycol dimethyl ether, triethyl orthoformate, formic anhydride, chloroformate, 1,1'-carbonyldiimidazole, or methanesulfonylimide salt. S4. In the presence of a diluent and an acid, compound VII is reacted with a 50% aqueous cyanamide solution at 70–140°C for 10–48 hours, and then recrystallized in a recrystallization reagent to obtain intermediate VIII; the diluent is the same as in step S1; the acid is an organic acid or an inorganic acid; the recrystallization reagent is an ether. S5. In the presence of alkali and a dehydrating agent, intermediate VI and intermediate VIII are reacted at 60–290°C for 10–60 hours, and the reaction solution is separated and purified to obtain intermediate IX; the alkali is the same as in step S1; the dehydrating agent is an alcohol compound. S6. In the presence of a diluent, an acid-binding agent, and a condensing agent, intermediate IV and intermediate IX are reacted at 0–50°C for 10–40 hours. The reaction solution is then separated and purified to obtain compound I. The diluent is the same as in step S1; the acid-binding agent is the same as in step S2; and the condensing agent is the same as in step S2. The R bases in V, VI, IX, and X are the same as those in I.

4. The preparation method according to claim 3, characterized in that, In step S1, the diluent is one of the following: benzene, toluene, xylene, chlorobenzene, dichlorobenzene, petroleum ether, hexane, cyclohexane, dichloromethane, chloroform, carbon tetrachloride, diethyl ether, diisopropyl ether, dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, acetone, butanone, methyl isobutyl ketone, acetonitrile, propionitrile, butyronitrile, N,N-dimethylformamide, N,N-dimethylacetamide, N-methylformaniline, N-methylpyrrolidone, hexamethylphosphoric triamine, methyl acetate, ethyl acetate, dimethyl sulfoxide, methanol, ethanol, 80-90% (v / v) aqueous ethanol solution, n-propanol, n-butanol, isopropanol, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, diethylene glycol monomethyl ether or diethylene glycol monoethyl ether; The alkali is sodium hydroxide, potassium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, sodium methoxide, sodium ethoxide, trimethylamine, pyridine, N,N-dimethylaniline, N,N-dimethylbenzylamine, N-methylpiperidine, or N-methylmorpholine; the catalyst is Pd(OAc)2, PdCl2, Pd(MeCN)2Cl2, Pd(PPh3)4, or Pd(TFA)2. The molar ratio of compound II to p-aminophenylborate acid salt is 1:1-1.5; the molar ratio of compound II to catalyst is 1:0.001-0.

1. The molar ratio of compound II to alkali is 1:2-5; the volume of diluent used is 1-5 mL / mmol based on the molar amount of compound II.

5. The method as described in claim 3, characterized in that, In step S2, the acid-binding agent is N,N-diisopropylethylamine, pyridine, N,N-dimethylaniline, N,N-dimethylbenzylamine, N-methylpiperidine, N-methylmorpholine, N,N-dimethylaminopyridine, diazabicyclooctane, diazabicyclononene or diazabicycloundecene, sodium hydroxide, sodium ethoxide, trimethylamine or triethylamine; The molar ratio of intermediate III to 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline hydrochloride is 1:1-1.5; the volumetric amount of the diluent is 1-5 mL / mmol based on the amount of intermediate III; the molar ratio of intermediate III to the acid-binding agent is 1:1-3; and the molar ratio of intermediate III to the condensing agent is 1:0.5-2.

6. The method as described in claim 3, characterized in that, In step S3, the ratio of compound V to the amount of acetal agent is 1:1-10.

7. The method as described in claim 3, characterized in that, In step S4, the acid is one or more of nitric acid, sulfuric acid, hydrochloric acid, phosphoric acid, p-toluenesulfonic acid, or trifluoroacetic acid; the recrystallization reagent is one or more of diethyl ether, tetrahydrofuran, methyl tert-butyl ether, isopropyl ether, or petroleum ether. The volumetric amount of the diluent used is 3-10 mL / mmol based on the amount of compound VII; the volumetric amount of the 50% cyanamide aqueous solution used is 0.1-1 mL / mmol based on the amount of compound VII; the molar ratio of compound VII to acid feed is 1:1-3; and the volumetric amount of the recrystallization reagent used for crystallization is 5-20 mL / mmol based on the amount of compound VII.

8. The method as described in claim 3, characterized in that, In step S5, the dehydrating agent is methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, sec-butanol, n-pentanol, isopentanol, benzyl alcohol, phenethyl alcohol, cyclohexanol, trifluoroethanol, 2-methoxyethanol, ethylene glycol, glycerol, or 1,2-propanediol. The molar ratio of intermediate VI to intermediate VIII is 1:1-3; the volumetric dosage of the dehydrating agent is 3-10 mL / mmol based on the amount of intermediate VI; and the molar ratio of intermediate VI to alkali is 1:2-5.

9. The method as described in claim 3, characterized in that, In step S6, the molar ratio of intermediate IV to intermediate IX is 1:1-1.5, the volume of diluent is 1-5 mL / mmol based on the amount of intermediate IV, the molar ratio of intermediate IV to acid-binding agent is 1:1-3, and the molar ratio of intermediate IV to condensing agent is 1:0.5-2.

10. The use of the N,4-diphenylpyrimidine-2-amine compound of claim 1 in the preparation of a tumor multidrug resistance reversal agent.

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