CN114605408A

CN114605408A - 5-hydroxy-1, 3-disubstituted phenyl pyrido [2,3-d ] pyrimidine compound and preparation method and application thereof

Info

Publication number: CN114605408A
Application number: CN202210322319.0A
Authority: CN
Inventors: 胡春; 谢倩; 李乐瑢; 王宇航; 文杰; 杨小力; 黄二芳; 彭子俊; 姚语桐; 林柔嘉; 金逸丹; 邓杰元
Original assignee: Shenyang Pharmaceutical University
Current assignee: Shenyang Pharmaceutical University
Priority date: 2022-03-30
Filing date: 2022-03-30
Publication date: 2022-06-10
Anticipated expiration: 2042-03-30
Also published as: CN114605408B

Abstract

5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] of the present invention]Pyrimidine compounds and a preparation method and application thereof, belonging to the technical field of medicines. In particular to 5-hydroxy-1, 3-disubstituted phenyl pyrido [2,3-d]The pyrimidine-2, 4,7(1H,3H,8H) -trione compound has a structural general formula shown in the formula (I): r₁Independently selected from hydrogen, fluoro, chloro, methyl, methoxy, trifluoromethyl, trifluoromethoxy; r₂Independently selected from hydrogen, methyl, butyl, cyclopropyl, 3-morpholinopropyl; r₃Independently selected from hydrogen, methyl, benzyl, 2-methylbenzyl. The compound has simple and convenient synthesis method, is suitable for industrial production, shows that the compound has anti-tumor activity through biological activity test, and can be applied to preparing anti-tumor drugs.

Description

5-hydroxy-1, 3-disubstituted phenyl pyrido [2,3-d ] pyrimidine compound and preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a 5-hydroxy-1, 3-disubstituted phenyl pyrido [2,3-d ] pyrimidine compound, and a preparation method and application thereof.

Background

Malignant tumor is a serious disease seriously threatening human health, has plagued human for many years, and is a big factor hindering the increase of human life span. With the discovery of oncogenes and the elucidation of cell signaling pathways, human understanding of the mechanism of canceration of cells has been greatly enriched, and by analyzing the function of oncogene products, many oncoproteins have been found to be located at different sites of normal cell signaling pathways, playing an important role in promoting cell division and proliferation (Klein G. science,1987(4833): 1539-.

The mitogen-activated protein kinase MAPK signaling pathway is a very important chain of information transfer in cells. The MAPK pathway has three stages of signal transmission processes: MAPK, MAPK kinases (MEK or MKK) and kinases of MAPK kinases (MEKK or mkkkk). These three kinases, which are sequentially activated and which together regulate many important physiological/pathological effects such as cell growth, differentiation, stress, inflammatory response, are the major areas of tumor development induction (Orton RJ, Sturm OE, Vyshemirsky V, Calder M, Gilbert DR, Kolch W. the Biochemical journal.2005,392(2): 249-61). The MAPK signal pathways commonly seen in human bodies mainly comprise a cell signal external kinase (ERK1/2) signal pathway, a c-Jun-N-terminal kinase (JNK)/Stress Activated Protein (SAPK) signal pathway, a p38MAPK signal pathway, mitogen activated extracellular signal-regulated kinase (MEK) and the like, and the MAPK pathway inhibitor small-molecule drugs which are on the market at present mainly comprise:

trametinib (Trametinib) is a MEK inhibitor drug with anticancer activity that inhibits the activity of MEK1 and MEK2 kinases. Trametinib produced good results in phase III clinical trials for metastatic melanoma, which carries the BRAF V600E mutation. In this mutation, the amino acid valine at position 600 of the BRAF protein has been substituted with glutamic acid, so that the mutated BRAF protein has constitutive activity (Flaherty KT, Infante JR, Daud A, Gonzalez R, Kefford RF, Sosman J, et al, the New England Journal of medicine.2012,367(18): 1694-.

Bimetinib (Binimetinib) is a selective inhibitor of MEK, developed by Array Biopharma for the treatment of various cancers. Binimetinib is a central kinase in the tumor-promoting MAPK pathway in MEK. Inappropriate activation of this pathway has been shown to occur in many cancers. 6 months 2018, it was approved by the FDA for use in combination with encorafenib in the treatment of patients with unresectable or metastatic BRAF V600E or V600K mutation-positive melanoma (Koelbinger P, Dornbierer J, Dummer R. future Oncology.2017,13(20): 1755-.

Sorafenib is an orally taken Raf kinase inhibitor, can directly inhibit Raf kinase activity, blocks signal transmission of a Ras pathway, and can also act on a platelet-derived growth factor receptor and a vascular endothelial growth factor receptor to down-regulate the expression level of p-MEK, thereby blocking a downstream Ras-Raf-MEK-ERK pathway. Sorafenib causes very low incidence of fatal adverse reactions such as gastrointestinal upset, skin toxicity, etc. (Wilhelm SM, Carter C, Tang L, et al cancer research.2004,64(19): 7099-.

Dalafinib (Dabraafinib) is a drug used for treating non-surgical or metastatic melanoma and can specifically act on B-Raf^V600EStructural domain, blocking MAPK pathway, inhibiting tumor cellsThe cell grows and proliferates, and the apoptosis of the cell is promoted. And researches show that the parallel passage causing the adverse reaction of the B-Raf inhibitor can be regulated and controlled by combining the B-Raf kinase inhibitor and the MEK inhibitor, and the combination of the B-Raf kinase inhibitor and the MEK inhibitor can improve the response speed and reduce the toxicity. Patients often develop resistance to dabrafenib more rapidly, and therefore this problem can be solved by combination with the MEK inhibitor trametinib (Khoja L, Hogg D.Extert Review of Anticancer therapy.2015,112(6): 536-545).

Disclosure of Invention

The invention aims to provide a 5-hydroxy-1, 3-disubstituted phenyl pyrido [2,3-d ] pyrimidine compound, in particular to a 5-hydroxy-1, 3-disubstituted phenyl pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione compound, a prodrug and a pharmaceutically acceptable salt thereof, a preparation method thereof and application thereof in preparing medicaments for treating or/and preventing tumor diseases related to MAPK pathways such as B-Raf kinase, Ras/Raf/MEK/ERK signal pathway, p38MAPK signal pathway, JNK-SAPK signal pathway and the like, and the application comprises application as a multi-target inhibitor.

5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidines as shown in formula I, prodrugs and pharmaceutically acceptable salts thereof;

R₁independently selected from hydrogen, halogen, C1-C4 alkyl, halogen substituted C1-C4 alkyl, C1-C4 alkoxy, halogen substituted C1-C4 alkoxy;

R₂independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, morpholinyl-substituted C1-C4 alkyl;

R₃independently selected from hydrogen, C1-C6 alkyl, benzyl, C1-C4 alkyl substituted benzyl.

Further, the compounds represented by formula I, prodrugs and pharmaceutically active metabolites thereof, and pharmaceutically acceptable salts thereof, according to the present invention:

R₁independently selected from hydrogenFluorine, chlorine, methyl, methoxy, trifluoromethyl, trifluoromethoxy;

R₂independently selected from hydrogen, methyl, butyl, cyclopropyl, 3-morpholinopropyl;

R₃independently selected from hydrogen, methyl, benzyl, 2-methylbenzyl.

Further, the following compounds are preferred in the present invention:

6-benzyl-8-butyl-5-hydroxy-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 01);

6-benzyl-5-hydroxy-8-methyl-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 02);

5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 03);

6-benzyl-5-hydroxy-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 04);

6-benzyl-5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 05);

6-benzyl-5-hydroxy-8-methyl-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 06);

6-benzyl-5-hydroxy-8-methyl-1, 3-bis [4- (trifluoromethyl) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 07);

6-benzyl-5-hydroxy-8-methyl-1, 3-bis [3- (trifluoromethyl) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 08);

6-benzyl-5-hydroxy-1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 09);

6-benzyl-1, 3-bis (4-chlorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 10);

6-benzyl-5-hydroxy-1, 3-bis (4-methoxyphenyl) -8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 11);

6-benzyl-5-hydroxy-8-methyl-1, 3-bis (2-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 12);

6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 13);

6-benzyl-8-butyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 14);

6-benzyl-8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 15);

6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8- (3-morpholinopropyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 16);

1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 17);

8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-6- (2-methylbenzyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 18);

1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methyl-6- (2-methylbenzyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 19);

6-benzyl-8-cyclopropyl-5-hydroxy-1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 20);

8-cyclopropyl-5-hydroxy-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 21);

5-hydroxy-8-methyl-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 22).

A pharmaceutical composition comprising as active ingredient a compound of any one of the compounds of the invention, prodrugs and pharmaceutically active metabolites thereof, and pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier or diluent.

"pharmaceutically acceptable salt" refers to conventional acid addition salts or base addition salts that retain the biological potency and properties of the compounds of formula I and are formed with non-toxic organic or inorganic acids or organic or inorganic bases. Acid addition salts include hydrochloride, hydrobromide, hydroiodide, nitrate, phosphate, sulfate, perchlorate, thiocyanate, bisulfate, persulfate, borate, formate, acetate, propionate, valerate, pivalate, hexanoate, heptanoate, octanoate, isooctanoate, undecanoate, laurate, palmitate, stearate, oleate, cyclopropionate, oxalate, malonate, succinate, maleate, fumarate, adipate, azelate, acrylate, strawberry, crotonate, glatironate, itaconate, sorbate, cinnamate, glycolate, lactate, malate, tartrate, citrate, tartrate, mandelate, diphenoxylate, troponate, ascorbate, gluconate, glucoheptonate, mandelate, dibenzolate, trogoplate, ascorbate, gluconate, glucoheptonate, and the like, Glucarate, mannonate, lactobionate, benzoate, phthalate, paraththalate, furoate, nicotinate, isonicotinate, salicylate, acetylsalicylate, caseinate, gallate, caffeate, ferulate, picrate, camphorate, camphorsulfonate, methanesulfonate, ethanesulfonate, propanesulfonate, benzenesulfonate, p-toluenesulfonate, sulfanilate, sulfamate, taurate, 2-hydroxyethanesulfonate, glycinate, alaninate, valine, leucine, isoleucine, phenylalanine, tryptophan, caseinate, aspartate, asparagine, glutamate, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine, arginine, and salts thereof, Edetate, pyruvate, alpha-ketoglutarate, alginate, cyclopentanepropionate, 3-phenylpropionate, 3-cyclohexylpropionic acid, 2-naphthoate, 2-naphthalenesulfonate, pamoate, lauryl sulfate, glycerophosphate, lauryl sulfate, pectin oleate. Preferred acids for the formation of acid addition salts include hydrochloric acid, p-toluenesulfonic acid, methanesulfonic acid, maleic acid, malic acid, picric acid, citric acid, sulfanilic acid. Base addition salts include ammonium salts, alkali metal salts, such as sodium and potassium salts, alkaline earth metal salts, such as calcium and magnesium salts, salts with organic bases, such as dicyclohexylamine salts, N-methyl-D-glucamine salts, and basic nitrogen-containing groups may be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and diamyl sulfate; long chain halides, such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides such as benzyl and phenethyl bromides.

The invention also relates to a pharmaceutical composition for inhibiting MEK kinase and B-RAF kinase, which comprises the compound shown in the formula I or the derivative or the pharmaceutically acceptable acid addition salt thereof and a pharmaceutically acceptable carrier.

"pharmaceutically acceptable" such as pharmaceutically acceptable carriers, excipients, prodrugs, means pharmacologically acceptable and substantially non-toxic to a patient to whom a particular compound is administered.

"pharmaceutically active metabolite" refers to a pharmaceutically acceptable and effective metabolite of a compound of formula I.

The term "halogen" as used in the present invention includes fluorine, chlorine, bromine and iodine.

The compounds of the present invention may be administered to a patient by various methods, including orally in capsules or tablets, as sterile solutions or suspensions, and in some cases, intravenously in the form of solutions. The free base compounds of the present invention may be formulated and administered in the form of their pharmaceutically acceptable acid addition salts.

The compound is used as a small molecule inhibitor related to MAPK signal pathways such as B-Raf kinase, Ras/Raf/MEK/ERK signal pathways, p38MAPK signal pathways, JNK-SAPK signal pathways and the like with a brand new structure type, has the characteristics of novel structure type, capability of acting on a plurality of targets and the like, and can be used for preparing and treating or preventing cancer diseases related to MAPK pathways such as B-Raf kinase, Ras/Raf/MEK/ERK signal pathways, p38MAPK signal pathways, JNK-SAPK signal pathways and the like; such as lung cancer, liver cancer, melanoma, colon cancer, rectal cancer, breast cancer, ovarian cancer, cervical cancer and renal cancer, and has good application value and development and application prospect.

The preparation route of the compound of the invention is as follows:

the compound XQA01-XQA22, pharmaceutically acceptable salts and prodrugs thereof can act on Ras/Raf/MEK/ERK signal pathways and inhibit phosphorylation of ERK1/2, thereby achieving the purpose of inhibiting tumor cell proliferation.

The invention has the beneficial effects that:

the invention provides a structure of a novel antitumor compound, has potential patent medicine value, simple and convenient preparation route, easy synthesis and low cost. Compared with the marketed drugs, the compound has more remarkable anti-tumor cell proliferation activity.

Detailed Description

The present invention is described in detail by the following examples. It should be understood, however, that the present invention is not limited to the following examples which are specifically set forth. Example 1: preparation of 6-benzyl-8-butyl-5-hydroxy-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA01)

Step I: weigh 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) into a 500mL round bottom flask, weigh p-toluidine (8.57g,80mmol) into 40mL methylene chloride, drop it into the flask with a dropping funnel at room temperature within 30min, weigh triethylamine (20.24g,0.2mol) into 50mL methylene chloride and drop it into the flask within 30min, weigh again p-toluidine (8.57g,80mmol) into 40mL methylene chloride, and drop it into the flask again as described above. After the dropping, the reaction was stirred for 30min to monitor the completion of the reaction of p-toluidine. And (3) distilling under reduced pressure to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to generate a large amount of solid, filtering out the solid, repeatedly washing with diethyl ether, and drying to obtain the 1, 3-di-p-toluidine as a white solid with the yield of 18.25g and the yield of 94.93%.

Step II: 1, 3-di-p-toluylurea (4.81g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring that the raw material 1, 3-di-p-toluidine urea completely reacts, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 1, 3-bis (4-methylphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellow-white solid 3.89g, in 63.08% yield.

Step III: 1, 3-bis (4-methylphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione (4.99g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 4.86g of 6-chloro-1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone, with the yield of 91.80%.

Step IV: 6-chloro-1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -dione (3.26g,10mmol), n-butylamine (1.09g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was stirred under reflux for 8H to complete the reaction. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6- (butylamino) -1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 3.46g and 95.20% yield.

Step V: weighing 6- (butylamino) -1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone (1.82g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-8-butyl-5-hydroxy-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] through column chromatography]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA01), white solid, yield 1.30g, 49.85% yield; m.p.: 85.5-88.9 ℃; MS 522.23676[ M ]+H]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.53(s,1H),7.40(d,J＝8.1Hz,2H),7.34(d,J＝8.2Hz,2H),7.29(d,J＝8.4Hz,3H),7.25(d,J＝2.9Hz,3H),7.22(d,J＝7.6Hz,2H),7.13(t,J＝6.9Hz,1H),3.47(t,J＝7.6Hz,2H),3.30(s,2H),2.36(d,J＝6.5Hz,6H),1.05(dq,J＝30.8,7.4Hz,2H),0.89–0.83(m,2H),0.68(t,J＝7.2Hz,3H)。

Example 2: preparation of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA 02):

the compound 6-chloro-1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -dione was prepared by the method of example 1.

Step IV: 6-chloro-1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -dione (3.26g,10mmol), 40% methylamine aqueous solution (1.28g,15mmol), triethylamine (2.22g,20mmol) and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6- (methylamino) -1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 3.16g, 98.33% yield. m.p.: 121.8-128.4 ℃; MS 480.18939[ M + H ]]⁺；

Step V: weighing 6- (methylamino) -1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone (1.61g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, carrying out reflux reaction for 40min, after TLC (thin layer chromatography) monitoring of the completion of the raw material reaction, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, carrying out suction filtration, drying, and carrying out column chromatography purification to obtain a pure product of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] of a pure product]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA02), white solid, yield 1.42g, yield 59.22%; m.p.: 121.8-128.4 ℃; MS 480.18939[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.43(s,1H),7.36(d,J＝8.5Hz,2H),7.32(d,J＝8.1Hz,3H),7.28(d,J＝2.9Hz,2H),7.25(d,J＝6.6Hz,2H),7.22(d,J＝7.9Hz,3H),7.16–7.11(m,1H),3.73(s,2H),2.73(s,3H),2.36(d,J＝1.9Hz,6H)。

Example 3: preparation of 5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA03)

Step IV: 6-chloro-1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -dione (3.26g,10mmol), N-aminopropylmorpholine (2.16g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6- [ (3-morpholinopropyl) amino ] -1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 3.16g and 72.72% yield.

Step V: weighing 6- [ (3-Morpholpropyl) amino group]Heating (2.17g,5mmol) of-1, 3-bis (4-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone (2.96 g,6mmol), diethyl malonate (0.96g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask on a heating jacket, refluxing and reacting for 40min, after TLC monitors that the raw materials are completely reacted, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (4-methylphenyl) pyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA03), white solid, yield 0.98g, yield 39.00%; m.p.: 209.0-212.2 ℃; MS 503.12516[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ11.96(s,1H),7.35–7.23(m,8H),6.06(s,1H),3.93(t,J＝6.8Hz,2H),3.53(t,J＝4.5Hz,4H),2.37(d,J＝3.9Hz,6H),2.24(t,J＝4.5Hz,4H),2.10(t,J＝7.2Hz,2H),1.59(p,J＝7.0Hz,2H)。

Example 4: preparation of 6-benzyl-5-hydroxy-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA04)

Step I: weigh 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) into a 500mL round bottom flask, weigh m-toluidine (8.57g,80mmol) into 40mL methylene chloride, drop it into the bottle with a dropping funnel within 30min at room temperature, weigh triethylamine (20.24g,0.2mol) into 50mL methylene chloride and drop it into the bottle within 30min, weigh again m-toluidine (8.57g,80mmol) into 40mL methylene chloride, and drop it into the bottle again as described above. After the dripping is finished, the reaction is stirred for 30min to monitor the m-toluidine reaction is complete. Distilling under reduced pressure to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to obtain a large amount of solid, filtering out the solid, washing with diethyl ether repeatedly, and drying to obtain 1, 3-di-toluurea as white solid with yield of 18.25g and 94.93%.

Step II: 1, 3-di-m-toluylurea (4.81g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring that the raw material 1, 3-di-p-toluidine urea completely reacts, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 1, 3-bis (3-methylphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellow-white solid 3.91g, 63.40% yield.

Step III: 1, 3-bis (3-methylphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione (4.99g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 4.92g of 6-chloro-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone, with the yield of 92.94%.

Step IV: 6-chloro-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -dione (3.26g,10mmol), 25% aqueous ammonia (2.1g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-amino-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in 2.93g yield 95.23%.

Step V: weighing 6-amino-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone (1.54g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] through column chromatography]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA04), white solid, yield 0.70g, 30.07% yield; m.p.: 134.7-138.4 ℃; MS 466.17416[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.36(s,1H),11.71(s,1H),7.34–7.40(m,2H),7.26–7.13(m,11H),3.77(s,2H),2.34(s,6H)。

Example 5: preparation of 6-benzyl-5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA05)

The compound 6-chloro-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -dione was prepared by the method of example 4.

Step IV: 6-chloro-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -dione (3.26g,10mmol), N-aminopropylmorpholine (2.16g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6- [ (3-morpholinopropyl) amino ] -1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 3.67g, 84.46% yield.

Step V: weighing 6- [ (3-Morpholpropyl) amino group]Heating (2.17g,5mmol) of-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone, diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask on a heating jacket to 256 ℃, refluxing and reacting for 40min, monitoring the reaction of raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether to precipitate a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (3-methylphenyl) pyriproxyfenPyrido [2,3-d]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA05), white solid, yield 0.64g, 21.60% yield; m.p.: 109.5-110.4 ℃; MS 593.27572[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.24(s,1H),7.36–7.40(m,2H),7.27–7.15(m,11H),3.92(t,J＝6.3Hz,2H),3.79(s,2H),3.51(t,J＝4.5Hz,4H),2.35(d,J＝3.3Hz,6H),2.18(s,4H),2.03(d,J＝7.4Hz,2H),1.54(p,J＝6.8Hz,2H)。

Example 6: preparation of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA06)

Step IV: 6-chloro-1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -dione (3.26g,10mmol), 40% methylamine aqueous solution (1.28g,15mmol), triethylamine (2.22g,20mmol) and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6- (methylamino) -1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in 3.06g, 95.23% yield.

And V: weighing 6- (methylamino) -1, 3-bis (3-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone (1.61g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, filtering, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] through column chromatography]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA06), white solid, yield 1.30g, yield 54.22%; m.p.: 111.6-118.3 ℃; MS 480.18945[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.41(s,1H),7.377.42(m,2H),7.20–7.35(m,9H),7.17–7.13(m,2H),3.72(d,J＝10.0Hz,2H),2.75(s,3H),2.34(s,6H)。

Example 7: preparation of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis [4- (trifluoromethyl) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA07)

Step I: weigh 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) into a 500mL round bottom flask, weigh p-trifluoromethylaniline (12.88g,80mmol) into 40mL of methylene chloride, drop it into the flask with a dropping funnel over 30min at room temperature, weigh triethylamine (20.24g,0.2mol) into 50mL of methylene chloride and drop it into the flask over 30min, weigh p-trifluoromethylaniline (12.88g,80mmol) again into 40mL of methylene chloride, and drop it into the flask again as described above. The reaction is stirred for 30min after the dripping is finished, and the p-fluoroaniline reaction is monitored to be complete. And (3) distilling under reduced pressure to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to generate a large amount of solid, filtering out the solid, repeatedly washing with diethyl ether, and drying to obtain the 1, 3-di-p-trifluoromethyl phenylurea as a white solid with the yield of 26.25g and the yield of 94.22%.

Step II: 1, 3-Di-p-trifluoromethylphenylurea (6.96g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring that the raw material 1, 3-di-p-toluidine urea completely reacts, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 6.41g of 1, 3-bis [4- (trifluoromethyl) phenyl ] pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellowish white solid in 76.99% yield.

Step III: 1, 3-bis [4- (trifluoromethyl) phenyl ] pyrimidine-2, 4,6(1H,3H,5H) -trione (6.74g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid 6-chloro-1, 3-bis [4- (trifluoromethyl) phenyl ] pyrimidine-2, 4(1H,3H) -diketone 6.92g with the yield of 98.26%.

Step IV: 6-chloro-1, 3-bis [4- (trifluoromethyl) phenyl ] pyrimidine-2, 4(1H,3H) -dione (4.35g,10mmol), 40% methylamine water solution (1.28g,15mmol), triethylamine (2.22g,20mmol) and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring under reflux for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-methylamino-1, 3-bis [4- (trifluoromethyl) phenyl ] pyrimidine-2, 4(1H,3H) -dione as a yellow solid in 4.09g yield 95.23%.

Step V: weighing 6-methylamino-1, 3-bis [4- (trifluoromethyl) phenyl ]]Putting pyrimidine-2, 4(1H,3H) -diketone (2.14g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring by TLC that the raw materials are completely reacted, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis [4- (trifluoromethyl) phenyl ] bis]Pyrido [2,3-d]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA07), white solid, yield 1.70g, yield 57.87%; m.p.: 123.2-128.3 ℃; MS 593.27572[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.24(s,1H),7.36–7.40(m,2H),7.27–7.15(m,11H),3.92(t,J＝6.3Hz,2H),3.79(s,2H),3.51(t,J＝4.5Hz,4H),2.35(d,J＝3.3Hz,6H),2.18(s,4H),2.03(d,J＝7.4Hz,2H),1.54(p,J＝6.8Hz,2H)。

Example 8: preparation of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis [3- (trifluoromethyl) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA08)

Step I: 40mL of methylene chloride was weighed to dissolve phosgene solid (9.5g,32mmol) in a 500mL round bottom flask, m-trifluoromethylaniline (12.88g,80mmol) was weighed to dissolve in 40mL of methylene chloride, it was dropped into the flask over 30min at room temperature using a dropping funnel, triethylamine (20.24g,0.2mol) was weighed to dissolve in 50mL of methylene chloride and dropped into the flask over 30min, m-trifluoromethylaniline (12.88g,80mmol) was weighed again to dissolve in 40mL of methylene chloride, and again dropped into the flask as described above. The reaction is stirred for 30min after the dripping is finished, and the p-fluoroaniline reaction is monitored to be complete. And (3) distilling under reduced pressure to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to generate a large amount of solid, filtering out the solid, repeatedly washing with diethyl ether, and drying to obtain the 1, 3-di-trifluoromethyl phenylurea as a white solid with the yield of 26.78g and the yield of 96.12%.

Step II: 1, 3-Bit-trifluoromethylphenylurea (6.96g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring that the raw material 1, 3-di-p-toluidine urea completely reacts, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 6.53g of 1, 3-bis [3- (trifluoromethyl) phenyl ] pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellowish white solid in 78.43% yield.

Step III: 1, 3-bis [3- (trifluoromethyl) phenyl ] pyrimidine-2, 4,6(1H,3H,5H) -trione (6.74g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid 6-chloro-1, 3-bis [3- (trifluoromethyl) phenyl ] pyrimidine-2, 4(1H,3H) -diketone 6.62g with the yield of 94.00%.

Step IV: 6-chloro-1, 3-bis [3- (trifluoromethyl) phenyl ] pyrimidine-2, 4(1H,3H) -dione (4.35g,10mmol), 40% methylamine water solution (1.28g,15mmol), triethylamine (2.22g,20mmol) and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring under reflux for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-amino-1, 3-bis [3- (trifluoromethyl) phenyl ] pyrimidine-2, 4(1H,3H) -dione as a yellow solid in 4.09g yield 95.35%.

Step V: weighing 6-methylamino-1, 3-bis[3- (trifluoromethyl) phenyl group]Putting pyrimidine-2, 4(1H,3H) -diketone (2.15g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring by TLC that the raw materials are completely reacted, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, filtering, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis [3- (trifluoromethyl) phenyl ] s]Pyrido [2,3-d]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA08), white solid, yield 1.56g, yield 53.10%; m.p.: 104.2-110.3 ℃; MS 588.13263[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.10(s,1H),7.93–7.67(m,8H),7.21–7.31(m,4H),7.17–7.11(m,1H),3.76(s,2H),2.76(s,3H)。

Example 9: preparation of 6-benzyl-5-hydroxy-1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA09)

Step I: 40mL of methylene chloride was weighed to dissolve phosgene solid (9.5g,32mmol) in a 500mL round bottom flask, p-trifluoroanisidine (14.17g,80mmol) was weighed to dissolve in 40mL of methylene chloride, which was dropped into the flask with a dropping funnel within 30min at room temperature, triethylamine (20.24g,0.2mol) was weighed to dissolve in 50mL of methylene chloride and dropped into the flask within 30min, p-trifluoroanisidine 14.17g,80mmol) was weighed again to dissolve in 40mL of methylene chloride, and again dropped into the flask as described above. The reaction is stirred for 30min after the dripping is finished, and the p-fluoroaniline reaction is monitored to be complete. And (3) distilling under reduced pressure to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to generate a large amount of solid, filtering out the solid, repeatedly washing with diethyl ether, and drying to obtain the 1, 3-di-p-trifluoromethoxy phenylurea as a white solid with the yield of 16.25g and the yield of 53.42%.

Step II: 1, 3-Di-p-trifluoromethoxybenzeneurea (7.60g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring the reaction completion of raw materials, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring to dissolve. The organic phase was extracted three times with 2mol/L NaOH aqueous solution, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellow-white solid 6.91g, 77.07% yield.

Step III: 1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrimidine-2, 4,6(1H,3H,5H) -trione (7.26g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 7.04g of 6-chloro-1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrimidine-2, 4(1H,3H) -diketone, with the yield of 93.11%.

Step IV: 6-chloro-1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrimidine-2, 4(1H,3H) -dione (4.67g,10mmol), 25% aqueous ammonia (2.1g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed out in a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-amino-1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrimidine-2, 4(1H,3H) -dione as a yellow solid in 4.26g yield 95.23%.

Step V: weighing 6-amino-1, 3-bis [4- (trifluoromethoxy) phenyl ]]Putting pyrimidine-2, 4(1H,3H) -diketone (2.24g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring by TLC that the raw materials are completely reacted, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, filtering, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-1, 3-bis [4- (trifluoromethoxy) phenyl]Pyrido [2,3-d]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA09), white solid, yield 0.86g, yield 28.41%; m.p.: 116.9 to 21.9 ℃; MS 606.10614[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.16(s,1H),11.76(s,1H),7.56–7.49(m,8H),7.20–7.25(m,4H),7.11–7.17(m,1H),3.78(s,2H)。

Example 10: preparation of 6-benzyl-1, 3-bis (4-chlorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA10)

Step I: 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) was weighed into a 500mL round bottom flask, p-chloroaniline (10.21g,80mmol) was weighed into 40mL of methylene chloride, which was dropped into the flask with a dropping funnel at room temperature within 30min, triethylamine (20.24g,0.2mol) was weighed into 50mL of methylene chloride and dropped into the flask within 30min, p-chloroaniline (10.21g,80mmol) was weighed again into 40mL of methylene chloride, and again dropped into the flask as described above. The reaction is stirred for 30min after the dripping is finished, and the p-fluoroaniline reaction is monitored to be complete. Vacuum distilling to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to produce a large amount of solid, filtering to remove solid, washing with diethyl ether repeatedly, and drying to obtain 1, 3-di-p-chlorophenylurea as white solid with yield of 10.12g and 45.00%.

Step II: 1, 3-Diphenylurea (5.62g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL of acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring the reaction completion of raw materials, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 1, 3-bis (4-chlorophenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellowish white solid 5.99g, in 85.78% yield.

Step III: 1, 3-bis (4-chlorophenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione (6.43g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol), and 20mL of acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 5.54g of 6-chloro-1, 3-bis (4-chlorophenyl) pyrimidine-2, 4(1H,3H) -diketone, with the yield of 93.11%.

Step IV: 6-chloro-1, 3-bis (4-chlorophenyl) pyrimidine-2, 4(1H,3H) -dione (3.68g,10mmol), 40% methylamine water solution (1.28g,15mmol), triethylamine (2.22g,20mmol), and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-methylamino-1, 3-bis (4-chlorophenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 3.46g and 95.52% yield.

Step V: weighing 6-methylamino-1, 3-bis (4-chlorophenyl) pyrimidine-2, 4(1H,3H) -dione (1.81g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, filtering, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-1, 3-bis (4-chlorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA10), white solid, yield 0.86g, 33.05% yield; m.p.: 131.3-150.4 ℃; MS 520.08032[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.23(s,1H),7.58–7.61(m,4H),7.52(d,J＝8.5Hz,2H),7.40(d,J＝8.6Hz,2H),7.30–7.20(m,4H),7.17–7.11(m,1H),3.74(s,2H),2.77(s,3H)。

Example 11: preparation of 6-benzyl-5-hydroxy-1, 3-bis (4-methoxyphenyl) -8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA11)

Step I: 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) was weighed into a 500mL round bottom flask, p-anisidine (9.82g,80mmol) was weighed into 40mL methylene chloride, which was dropped into the flask with a dropping funnel within 30min at room temperature, triethylamine (20.24g,0.2mol) was weighed into 50mL of methylene chloride and into the flask within 30min, p-anisidine (9.82g,80mmol) was weighed again into 40mL of methylene chloride, and again into the flask as described above. The reaction is stirred for 30min after the dripping is finished, and the p-fluoroaniline reaction is monitored to be complete. And (3) distilling under reduced pressure to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to generate a large amount of solid, filtering out the solid, repeatedly washing with diethyl ether, and drying to obtain the 1, 3-di-p-methoxyphenylurea as a white solid with the yield of 10.57g and the yield of 48.52%.

Step II: 1, 3-di-p-methoxyphenylurea (5.45g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring the reaction completion of raw materials, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 1, 3-bis (4-methoxyphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellowish white solid 5.01g with a yield of 73.60%.

Step III: 1, 3-bis (4-methoxyphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione (5.51g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 5.41g of 6-chloro-1, 3-bis (4-methoxyphenyl) pyrimidine-2, 4(1H,3H) -diketone, with the yield of 93.11%.

Step IV: 6-chloro-1, 3-bis (4-methoxyphenyl) pyrimidine-2, 4(1H,3H) -dione (3.59g,10mmol), 40% methylamine aqueous solution (1.28g,15mmol), triethylamine (2.22g,20mmol) and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-methylamino-1, 3-bis (4-methoxyphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in 3.37g yield 95.23%.

Step V: 6-methylamino-1, 3-bis (4-methoxyphenyl) pyrimidine-2, 4(1H,3H) -dione (1.77g,5mmol), diethyl benzylmalonate (1.50g,6 m) were weighedmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating the mixture to 256 ℃ on a heating jacket, carrying out reflux reaction for 40min, monitoring the reaction of the raw materials by TLC (thin layer chromatography), pouring the reaction solution into 30mL petroleum ether, separating out a large amount of solid, carrying out suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-1, 3-bis (4-methoxyphenyl) -8-methylpyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA11), white solid, yield 1.02g, 39.88% yield; m.p.: 104.1-111.2 ℃; MS 512.17969[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.47(s,1H),7.42–7.37(m,2H),7.30–7.22(m,6H),7.16–7.11(m,1H),7.08–7.01(m,4H),3.80(s,3H),3.80(s,3H),3.73(s,2H),2.75(s,3H)。

Example 12: preparation of 6-benzyl-5-hydroxy-8-methyl-1, 3-di-o-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA12)

Step I: weigh 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) into a 500mL round bottom flask, weigh o-toluidine (8.57g,80mmol) into 40mL methylene chloride, drop it into the flask with a dropping funnel at room temperature within 30min, weigh triethylamine (20.24g,0.2mol) into 50mL methylene chloride and drop it into the flask within 30min, weigh again o-toluidine (8.57g,80mmol) into 40mL methylene chloride, and drop it into the flask again as described above. After the dripping is finished, the reaction is stirred for 30min to monitor the reaction completion of the o-toluidine. Vacuum distilling to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to produce a large amount of solid, filtering out the solid, washing with diethyl ether repeatedly, and drying to obtain 1, 3-di-o-toluidine, white solid, 10.57g yield, 54.98% yield.

Step II: 1, 3-Di-o-tolylurea (4.81g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring the reaction completion of raw materials, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 5.01g of 1, 3-bis (2-methylphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellowish white solid in 81.24% yield.

Step III: 1, 3-bis (2-methylphenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione (5.00g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 4.93g of 6-chloro-1, 3-bis (2-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone, with the yield of 93.11%.

Step IV: 6-chloro-1, 3-bis (2-methylphenyl) pyrimidine-2, 4(1H,3H) -dione (3.27g,10mmol), 40% methylamine aqueous solution (1.28g,15mmol), triethylamine (2.22g,20mmol) and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-methylamino-1, 3-bis (2-methylphenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 3.06g, 95.23% yield.

Step V: weighing 6-methylamino-1, 3-bis (2-methylphenyl) pyrimidine-2, 4(1H,3H) -diketone (1.61g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, filtering, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-5-hydroxy-8-methyl-1, 3-bis (2-methylphenyl) pyrido [2,3-d ] after the reaction is completed, and purifying by column chromatography]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA12), white solid, yield 1.02g, yield 42.54%; m.p.: 97.0 to 108.4 ℃; MS 480.18936[ M + H ]]⁺；¹H NMR(400MHz,)δ12.51(s,1H),7.47(d,J＝7.8Hz,1H),7.35-7.44m,4H),7.23(dd,J＝8.4,6.6Hz,5H),7.13–7.15(m,2H),2.66(s,2H),2.64(s,3H),2.30(s,6H)。

Example 13: preparation of 6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA13)

Step I: 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) was weighed into a 500mL round-bottom flask, para-fluoroaniline (8.89g, 80mmol) was weighed into 40mL of methylene chloride, which was dropped into the flask with a dropping funnel over 30min at room temperature, triethylamine (20.24g,0.2mol) was weighed into 50mL of methylene chloride and dropped into the flask over 30min, and para-fluoroaniline (8.89g, 80mmol) was weighed into 40mL of methylene chloride and again dropped into the flask as described above. The reaction is stirred for 30min after the dripping is finished, and the p-fluoroaniline reaction is monitored to be complete. And (3) carrying out reduced pressure distillation to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to generate a large amount of solid, filtering out the solid, repeatedly washing with diethyl ether, and drying to obtain the 1, 3-di-p-fluorophenylurea as a white solid with the yield of 18.5g and the yield of 93.16%.

Step II: 1, 3-Di-p-fluorophenylurea (4.96g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring the reaction completion of raw materials, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 1, 3-bis (4-fluorophenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione as a yellow-white solid 3.87g, yield 61.18%.

Step III: 1, 3-bis (4-fluorophenyl) pyrimidine-2, 4,6(1H,3H,5H) -trione (5.12g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 5.12g of 6-chloro-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -diketone, with the yield of 94.43%.

Step IV: 6-chloro-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (3.35g,10mmol), 40% methylamine water solution (1.28g,15mmol), triethylamine (2.22g,20mmol) and 20mL ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-methylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 3.28g and a yield of 99.60%.

Step V: weighing 6-methylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -diketone (1.65g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, filtering, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA16), white solid, yield 1.21g, 49.64% yield; m.p.: 136.4-148.7 ℃; MS 510.12143[ M + Na ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.31(s,1H),7.59–7.52(m,2H),7.44–7.32(m,6H),7.29–7.20(m,4H),7.17–7.11(m,1H),3.74(s,2H),2.76(s,3H)。

Example 14: preparation of 6-benzyl-8-butyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA14)

The compound 6-chloro-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione was obtained by the method of example 13.

Step IV: 6-chloro-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (3.35g,10mmol), n-butylamine (1.09g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-butylamine-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 2.48g and 82.66% yield.

Step V: 6-Butylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (1.86g,5mmol), benzyl propyl ester were weighedDiethyl diacid (1.50g,6mmol) and 15g diphenyl ether are put in a 100mL three-neck round-bottom flask, heated to 256 ℃ on a heating jacket and then refluxed for 40min, TLC monitors that the raw materials are completely reacted, the reaction liquid is poured into 30mL petroleum ether, a large amount of solid is separated out, filtered, dried and purified by column chromatography to obtain a pure product of 6-benzyl-8-butyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA17), white solid, yield 1.03g, 38.90% yield; m.p.: 104.8 to 107.1 ℃; MS 530.18634[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.42(s,1H),7.64–7.56(m,2H),7.47–7.38(m,4H),7.37–7.31(m,2H),7.27–7.21(m,4H),7.17–7.10(m,1H),3.75(s,2H),3.48(t,J＝7.6Hz,2H),1.10–1.03(m,2H),0.89–0.93(m,2H),0.70(t,J＝7.2Hz,3H)。

Example 15: preparation of 6-benzyl-8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA15)

Step IV: 6-chloro-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (3.35g,10mmol), cyclopropylamine (0.85g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-cyclopropylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 2.77g and 77.95% yield.

Step V: weighing 6-cyclopropylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (1.78g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, carrying out reflux reaction for 40min, monitoring the reaction of raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, carrying out suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA18), white solid, yield 0.89g, yield 34.66%; m.p.: 225.9-28.2℃；MS:514.15637[M+H]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.17(s,1H),7.54(dd,J＝8.8,4.8Hz,2H),7.42(dd,J＝8.8,5.1Hz,2H),7.39–7.18(m,8H),7.14(dd,J＝8.0,5.0Hz,1H),3.71(s,2H),1.64(p,J＝5.4Hz,1H),0.80(d,J＝6.0Hz,4H)。

Example 16: preparation of 6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8- (3-morpholinopropyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA16)

Step IV: 6-chloro-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (3.35g,10mmol), N-aminopropylmorpholine (2.16g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-morpholinopropylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 2.32g and 54.15% yield.

Step V: weighing 6-morpholine propylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -diketone (2.14g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8- (3-morpholine propyl) pyrido [2,3-d ] after the reaction is completed]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA19), white solid, yield 1.03g, 34.30% yield; m.p.: 181.3 to 181.6 ℃; MS 601.22611[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.14(s,1H),7.41–7.55(m,4H),7.29–7.40(m,4H),7.17–7.27(m,4H),7.11–7.17(m,1H),3.93(t,J＝6.5Hz,2H),3.80(s,2H),3.51(t,J＝4.5Hz,4H),2.18(t,J＝4.6Hz,4H),2.05(t,J＝7.2Hz,2H),1.56(p,J＝6.8Hz,2H)。

Example 17: preparation of 1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA17)

The compound 6-methylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione was prepared by the method of example 13.

Step V: weighing 6-methylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -diketone (1.65g,5mmol), diethyl malonate (0.96g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product 1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA20), white solid, yield 1.39g, 69.97% yield; m.p.: 246.3 to 253.6 ℃; MS 398.09290[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.00(s,1H),7.60–7.54(m,2H),7.43–7.34(m,6H),5.63(s,1H),2.73(s,3H)。

Example 18: preparation of 8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-6- (2-methylbenzyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA18)

Step IV: 6-chloro-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (3.34g,10mmol), cyclopropylamine (0.85g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-cyclopropylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione as a yellow solid in a yield of 2.77g, 77.95% yield.

Step V: weighing 6-cyclopropylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -dione (1.78g,5mmol), diethyl 2- (2-methylbenzyl) malonate (1.58g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, carrying out reflux reaction for 40min, monitoring the reaction of raw materials by TLC (thin layer chromatography), pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, carrying out suction filtration, drying, and purifying by column chromatography to obtain a pure product8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-6- (2-methylbenzyl) pyrido [2,3-d]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA21), white solid, yield 0.95g, yield 36.02%; m.p.: 133.2-140.6 ℃; MS 528.17218[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.16(s,1H),7.62–7.51(m,2H),7.48–7.40(m,2H),7.39–7.28(m,4H),7.16–7.09(m,1H),7.09–6.95(m,3H),3.65(s,2H),2.36(s,3H),1.63-1.67(m,1H),0.81(dd,J＝12.1,6.0Hz,4H)。

Example 19: preparation of 1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methyl-6- (2-methylbenzyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA19)

Step V: weighing 6-methylamino-1, 3-bis (4-fluorophenyl) pyrimidine-2, 4(1H,3H) -diketone (1.65g,5mmol), diethyl 2- (2-methylbenzyl) malonate (1.58g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methyl-6- (2-methylbenzyl) pyrido [2,3-d ] pyridine [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA22), white solid, yield 0.85g, yield 33.90%; m.p.: 118.8-125.3 ℃; MS 502.15601[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.30(s,1H),7.62–7.54(m,2H),7.46–7.32(m,6H),7.08–7.16(m,1H),7.07–7.01(m,3H),3.69(s,2H),2.77(s,3H),2.37(s,3H)。

Example 20: preparation of 6-benzyl-8-cyclopropyl-5-hydroxy-1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA20)

Step I: weigh 40mL of methylene chloride dissolved phosgene solid (9.5g,32mmol) into a 500mL round bottom flask, weigh aniline (7.45g,80mmol) into 40mL methylene chloride, drop it into the flask with a dropping funnel at room temperature over 30min, weigh triethylamine (20.24g,0.2mol) into 50mL methylene chloride and drop it into the flask over 30min, weigh again aniline (7.4g,80mmol) into 40mL methylene chloride and drop it into the flask again as described above. The reaction is stirred for 30min after the dripping is finished, and the p-fluoroaniline reaction is monitored to be complete. And (3) carrying out reduced pressure distillation to remove about 3/4 volume of dichloromethane, stirring with 5% HCl solution for 5-10min to generate a large amount of solid, filtering out the solid, repeatedly washing with diethyl ether, and drying to obtain the 1, 3-diphenyl urea as a white solid with the yield of 16.17g and the yield of 93.25%.

Step II: 1, 3-diphenylurea (4.25g,20mmol), malonic acid (2.7g,26mmol), phosphorus oxychloride (3.4g,22mmol) and 15mL acetonitrile were weighed into a35 mL sealed tube. Sealing and heating to 120 ℃, reacting for 1h, monitoring the reaction completion of raw materials, pouring the reaction liquid in the sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding 50mL of 2mol/L NaOH and 20mL of ethyl acetate, and stirring and dissolving. The organic phase was extracted three more times with 2mol/L aqueous NaOH, the aqueous layers were combined, the aqueous layer was extracted once with a small amount of ethyl acetate, and the remaining aqueous layer was adjusted to pH with hydrochloric acid to give a large amount of solid. The solid was filtered off, the filter cake was washed with water and dried to give 1, 3-diphenylpyrimidine-2, 4,6(1H,3H,5H) -trione as a yellowish white solid 3.75g, in 66.90% yield.

Step III: 1, 3-diphenylpyrimidine-2, 4,6(1H,3H,5H) -trione (4.54g,16.2mmol), phosphorus oxychloride (4.92g,32.4mmol) and 20mL of acetonitrile were weighed into a 125mL sealed tube. Sealing and heating to 120 ℃ for reaction for 3H, monitoring the reaction completion of raw materials, pouring the reaction liquid in a sealed tube into an eggplant-shaped bottle, distilling under reduced pressure to remove all solvents, adding a small amount of ethanol to dissolve the mixture, adding water to precipitate a solid, filtering out the solid, washing a filter cake with water, and drying to obtain a yellow solid, namely 4.58g of 6-chloro-1, 3-diphenylpyrimidine-2, 4(1H,3H) -diketone, with the yield of 94.64%.

Step IV: 6-chloro-1, 3-diphenylpyrimidine-2, 4(1H,3H) -dione (2.99g,10mmol), cyclopropylamine (1.10g,15mmol), triethylamine (2.22g,20mmol) and 20mL of ethanol were weighed into a 50mL round-bottomed flask, and the reaction was completed after stirring and refluxing for 8 hours. Most of the ethanol was distilled off under reduced pressure, and diethyl ether was added to stir the solid out. The solid was filtered off, washed with diethyl ether and dried to give 6-cyclopropylamino-1, 3-diphenylpyrimidine-2, 4(1H,3H) -dione as a yellow solid in 2.54g yield 79.53%.

Step V: weighing 6-cyclopropylamino-1, 3-diphenylpyrimidine-2, 4(1H,3H) -diketone (1.60g,5mmol), diethyl benzylmalonate (1.50g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, carrying out reflux reaction for 40min, after TLC (thin layer chromatography) monitors that raw materials are completely reacted, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, carrying out suction filtration, drying, and purifying by column chromatography to obtain a pure product of 6-benzyl-8-cyclopropyl-5-hydroxy-1, 3-diphenylpyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA35), white solid, yield 0.85g, 35.60% yield; m.p.: 238.2 to 241.3 ℃; MS 478.17505[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.24(s,1H),7.53–7.43(m,8H),7.40–7.35(m,2H),7.27–7.21(m,4H),7.17–7.11(m,1H),3.71(s,2H),1.58–1.49(m,1H),0.84–0.73(m,4H)。

Example 21: preparation of 8-cyclopropyl-5-hydroxy-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA21)

The compound 6-cyclopropylamino-1, 3-diphenylpyrimidine-2, 4(1H,3H) -dione was obtained by the method of example 35.

Step V: weighing 6-cyclopropylamino-1, 3-diphenylpyrimidine-2, 4(1H,3H) -dione (1.60g,5mmol), diethyl 2- (2-methylbenzyl) malonate (1.58g,6mmol) and 15g diphenyl ether in a 100mL three-necked round-bottom flask, heating to 256 ℃ on a heating jacket, carrying out reflux reaction for 40min, after TLC (thin-layer chromatography) monitors that the raw materials are completely reacted, pouring the reaction liquid into 30mL petroleum ether, separating out a large amount of solid, carrying out suction filtration, drying, and purifying by column chromatography to obtain a pure product of 8-cyclopropyl-5-hydroxy-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA36), white solid, yield 0.92g, yield 37.43%; m.p.: 276.6-279.3℃；MS:492.19162[M+H]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.24(s,1H),7.55–7.43(m,8H),7.41–7.37(m,2H),7.16–7.09(m,1H),7.08–6.96(m,3H),3.66(s,2H),2.36(s,3H),1.55(tt,J＝7.0,4.0Hz,1H),0.92–0.82(m,2H),0.80–0.68(m,2H)。

Example 22: preparation of 5-hydroxy-8-methyl-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA22)

The compound 6-methylamino-1, 3-diphenylpyrimidine-2, 4(1H,3H) -dione was obtained by the method of example 30.

Step V: weighing 6-methylamino-1, 3-diphenylpyrimidine-2, 4(1H,3H) -diketone (1.47g,5mmol), diethyl 2- (2-methylbenzyl) malonate (1.58g,6mmol) and 15g diphenyl ether in a 100mL three-neck round-bottom flask, heating to 256 ℃ on a heating jacket, refluxing for 40min, monitoring the reaction of the raw materials by TLC, pouring the reaction liquid into 30mL petroleum ether, precipitating a large amount of solid, performing suction filtration, drying, and purifying by column chromatography to obtain a pure product of 5-hydroxy-8-methyl-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ]]Pyrimidine-2, 4,7(1H,3H,8H) -trione (XQA37), white solid, yield 1.01g, yield 43.39%; m.p.: 113.9-140.3 ℃; MS 466.17529[ M + H ]]⁺；¹H NMR(400MHz,DMSO-d₆)δ12.38(s,1H),7.57–7.43(m,8H),7.41–7.35(m,2H),7.15–7.09(m,1H),7.07–7.01(m,3H),3.69(s,2H),2.73(s,3H),2.37(s,3H)。

Pharmacological examples

Example 23: inhibitory Activity of test Compounds on MCF7, A375, SK-Mel-2 cell proliferation

(1) Experimental Material

Cell line (b): MCF7, A375 and SK-Mel-2 cells were plated in 96-well plates at densities of 4000, 5000 and 4000/well, 200 ul/well, and used after 24 h.

No. XQA01-XQA22 target compound: dissolved in DMSO, diluted with culture medium to obtain six different concentrations of 100. mu.M, 50. mu.M, 25. mu.M, 12.5. mu.M, 6.25. mu.M and 3.125. mu.M, and stored at-20 deg.C for use, with the final concentration of DMSO in the culture medium being less than 0.1%.

Positive control drug: 5-Fluorouracil (5-Fu, Fluorouracil).

MTT: dissolved in PBS to 2mg/mL and stored at-20 ℃.

(2) Experimental method

MCF7, A375, SK-Mel-2 cells were selected to evaluate the anti-tumor proliferation activity of test samples using the MTT method. MCF7, A375, SK-Mel-2 cell lines were cultured in DMEM medium containing 10% bovine serum (FBS). Cells were pooled when they proliferated to 80-90% and subsequently subcultured for no more than 20 passages, and then allowed to acclimate for 24h before further processing. These cells were plated in 96-well plates and then incubated in a medium containing 5% CO₂Culturing in an incubator at constant temperature of 37 ℃ until the cells are completely attached to the wall. After 24h, various concentrations of representative compounds of the invention were added. After an additional 24h of incubation, MTT (2mg/mL) was added and incubation continued for 4 h. The culture medium was removed, the crystals were dissolved in DMSO, and the absorbance was measured at a wavelength of 570nm using a microplate reader (Thermo Multiskan GO, Thermo Fisher, USA). According to the formula: the cell growth inhibition rate is (1-drug group OD value/control group OD value) × 100%, the cell growth inhibition rate under the corresponding concentration is calculated, and the IC corresponding to the tested compound is calculated according to the logarithmic curve of the inhibition rate of the tested compound to the cell and the different concentrations of the tested compound₅₀The value is obtained. Representative compounds of the invention were tested according to the methods described above.

Most of the 22 compounds prepared above showed moderate tumor cell proliferation inhibiting activity on MCF-7, A375 and SK-MEL-2 cell lines, and XQA09 showed superior proliferation inhibiting activity to the positive control drug, IC₅₀As low as 9.92. + -. 0.52. mu.M and 8.80. + -. 0.53. mu.M.

Formulation examples

The following formulation examples are merely illustrative of the scope of the invention and are not to be construed as limiting in any way. The active compounds described in the following examples refer to compounds XQA01 to XQA22 prepared in the above examples.

Example 24: tablet formulation

25-1000mg of active compound, 45mg of starch, 35mg of microcrystalline cellulose, 4mL of polyvinylpyrrolidone (as a 10% aqueous solution), 4.5mg of sodium carboxymethylcellulose, 0.5mg of magnesium stearate, and 1mg of talc.

Example 25: suspending agent formula

0.1-1000mg of active compound, 50mg of sodium carboxymethylcellulose, 1.25mg of syrup, 0.1mg of sodium benzoate, 25mg of flavoring agent and 5mg of coloring agent, and pure water is added to the volume of 5 mL.

Example 26: aerosol formulations

0.25mg of active compound, 25-75mL of ethanol and 70mg of propellant 22 (chlorodifluoromethane).

Example 27: suppository formula

250mg of active compound, 2000mL of saturated fatty acid glycerides.

Example 28: injectable formulation

50mg of active compound, 1000mL of isotonic saline solution.

Example 29: ointment formulation

0.025g of micronized active compound, 10g of liquid paraffin, and 100g of soft white wax.

Example 30: ointment formulation

0.025g of active compound, 5g of propylene glycol, 5g of sorbitan sesquioleate, 10g of liquid paraffin and 100g of soft white wax.

Example 31: oil-in-water cream formulation

0.025g of active compound, 5g of cetyl alcohol, 5g of glycerol monostearate, 10g of liquid paraffin, 2g of cetyl polyoxyethylene ether, 0.1g of citric acid, 0.2g of sodium citrate, 35g of propylene glycol and water to 100 g.

Example 32: oil-in-water cream formulation

0.025g of micronized active compound, 15g of soft white wax, 5g of liquid paraffin, 5g of cetyl alcohol, 2g of Sorbimcarol stearate (Tween 65 of the particular pharmaceutical excipient grade), 0.5g of sorbitan monostearate, 0.2g of sorbic acid, 0.1g of citric acid, 0.2g of sodium citrate, and water to 100 g.

Example 33: water-in-oil cream formulation

0.025g of active compound, 35g of soft white wax, 5g of liquid paraffin, 5g of sorbitan sesquioleate, 0.2g of sorbic acid, 0.1g of citric acid and 0.2g of sodium citrate, and water is added until the weight is 100 g.

Example 34: lotion formulation

0.25g of active compound, 0.5mL of isopropanol, 3mg of carboxyvinyl polymer, 2mg of NaOH and water to 1 g.

Example 35: injectable suspension formulations

10mg of the active compound, 7mg of sodium carboxymethylcellulose, 7mg of NaCl, 0.5mg of polyoxyethylene (20) sorbitan monooleate, 8mg of benzyl alcohol, and sterile water to 1 mL.

Example 36: aerosol formulation for oral and nasal inhalation

0.1% w/w active compound, 0.7% w/w sorbitan trioleate, 24.8% w/w trichlorofluoromethane, 24.8% w/w dichlorotetrafluoroethane and 49.6% w/w dichlorodifluoromethane.

Example 37: atomized solution formulation

7mg of active compound, 5mg of propylene glycol, water to 10 g.

Example 38: powder formulations for inhalation

Gelatine capsules were filled with a mixture of the following ingredients, 0.1mg of micronised active compound, 20mg of lactose and the powder was inhaled by means of an inhalation device.

Example 39: powder formulations for inhalation

The spheronized powder was loaded into a multi-dose powder inhaler containing 0.1mg of micronized active compound per dose.

Example 40: powder formulations for inhalation

The spheronized powder was loaded into a multi-dose powder inhaler containing 0.1mg of micronized active compound and 1mg of micronized lactose per dose.

Example 41: capsule formulation

1.0mg of active compound, 321mg of small sugar spheres, 306.6 mg of Aquacoat ECD, 0.5mg of acetyl tributyl citrate, 800.1 mg of Tween-800, 100-5517.5 mg of Eudragit L, 1.8mg of triethyl citrate, 8.8mg of talcum powder and 0.lmg of defoamer MMSg.

Example 42: capsule seedling formula

2.0mg of active compound, 305mg of small sugar spheres, Aquocoat ECD 305.0 mg, acetyl tributyl citrate 0.4mg, Tween-800.14 mg, Eudragit NE 30D 12.6mg, Eudragit S10012.6 mg, talc 0.l6 mg.

Example 43: enema formula

2mg of active compound, 25mg of sodium carboxymethylcellulose, 0.5mg of disodium ethylenediaminetetraacetate, 0.8mg of methylparaben, 0.2mg of propylparaben, 7mg of sodium chloride, 1.8mg of citric acid, 800.01 mg of tween-tween and 1mL of pure water.

Example 44: formulations containing liposomes

A. Preparation of a drip infusion formula:

dipalmitoyl lecithin (45mg), dimyristoyl lecithin (7mg), dipalmitoyl phosphatidyl glycerol (1mg) and active compound (5mg) were placed in a glass tube, all components were dissolved in chloroform and N was used₂The pressure was reduced after evaporating most of the solvent, whereby a thin lipid film was formed on the surface of the glass tubeLiposomes were formed at a temperature above the phase inversion temperature of the lipids by adding an aqueous solution (0.9% NaCl) to the lipids, and the resulting suspension contained liposomes ranging in size from very small vesicles to 2 μm.

B. Preparation of formulation for inhalation:

liposomes were prepared as in example A, with an aqueous solution containing 10% lactose at a 7:3 lactose to lipid ratio. The liposome suspension was frozen with dry ice and freeze-dried, and the dried product was micronized, and the Mass Mean Aerodynamic Diameter (MMAD) of the resulting particles was about 2 μm.

Claims

1. A 5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidines compound of formula I, prodrugs and pharmaceutically active metabolites thereof, and pharmaceutically acceptable salts thereof, wherein the formula I has the structure:

R₂independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, morpholinyl substituted C1-C4 alkyl;

2. The 5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidines, prodrugs and pharmaceutically active metabolites thereof, and pharmaceutically acceptable salts thereof according to claim 1, wherein in formula I:

R₁independently selected from hydrogen, fluoro, chloro, methyl, methoxy, trifluoromethyl, trifluoromethoxy;

R₃independently selected from hydrogen, AA benzyl group, a 2-methylbenzyl group.

3. The 5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidines, prodrugs and pharmaceutically active metabolites thereof, and pharmaceutically acceptable salts thereof according to claim 2, wherein the compound is any one of the following compounds:

6-benzyl-8-butyl-5-hydroxy-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-8-methyl-1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (4-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-8- (3-morpholinopropyl) -1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-8-methyl-1, 3-bis (3-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-8-methyl-1, 3-bis [4- (trifluoromethyl) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-8-methyl-1, 3-bis [3- (trifluoromethyl) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-1, 3-bis [4- (trifluoromethoxy) phenyl ] pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-1, 3-bis (4-chlorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-1, 3-bis (4-methoxyphenyl) -8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-5-hydroxy-8-methyl-1, 3-bis (2-methylphenyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-8-butyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxypyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-8- (3-morpholinopropyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

8-cyclopropyl-1, 3-bis (4-fluorophenyl) -5-hydroxy-6- (2-methylbenzyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

1, 3-bis (4-fluorophenyl) -5-hydroxy-8-methyl-6- (2-methylbenzyl) pyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

6-benzyl-8-cyclopropyl-5-hydroxy-1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

8-cyclopropyl-5-hydroxy-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione;

5-hydroxy-8-methyl-6- (2-methylbenzyl) -1, 3-diphenylpyrido [2,3-d ] pyrimidine-2, 4,7(1H,3H,8H) -trione.

4. A pharmaceutical composition comprising as an active ingredient a compound of any one of the 5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidines of claims 1-3, prodrugs and pharmaceutically active metabolites thereof, and pharmaceutically acceptable salts thereof, and a pharmaceutically acceptable carrier or diluent.

5. A pharmaceutical composition according to claim 4, which is a pharmaceutical composition for inhibiting mitogen-activated protein kinase.

6. A process for the preparation of 5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidines, prodrugs and pharmaceutically active metabolites thereof, and pharmaceutically acceptable salts thereof according to claim 3, wherein the process comprises the following steps:

7. use of a 5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidine according to any one of claims 1 to 3, a prodrug and a pharmaceutically active metabolite thereof, a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to any one of claims 4 to 5 for the preparation of a mitogen-activated protein kinase, a mitogen-activated protein kinase, an extracellular regulated protein kinase, a Raf protein kinase inhibitor.

8. Use of a 5-hydroxy-1, 3-disubstituted phenylpyrido [2,3-d ] pyrimidine according to any one of claims 1 to 3, a prodrug and a pharmaceutically active metabolite thereof, a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to any one of claims 4 to 5 for the manufacture of a medicament for the treatment of cancer.

9. The use of claim 8, wherein the cancer is lung cancer, liver cancer, melanoma, colon cancer, rectal cancer, breast cancer, ovarian cancer, cervical cancer, or renal cancer.