1-aryl-3-(heterocyclic substituted phenyl) urea compound as well as preparation method and application thereof

Abstract

The invention discloses 1-aryl-3-(heterocyclic substituted phenyl) urea compounds as well as a preparation method and application thereof, and belongs to the technical field of medicines. In particular to 1-aryl-3-{4-{1-[(3aR, 4R, 8R, 8aR)-2, 2, 6-trimethylhexahydro-5H-4, 8-epoxy [1, 3] dioxolane [4, 5-d] [1, 2] diaza # imgabs0 #-5-yl] methyl} phenyl} urea compounds as well as a preparation method and application thereof in preparation of antitumor drugs. The structural general formula of the 1-aryl-3-(heterocyclic substituted phenyl) urea compound is shown in the specification, wherein Y is CH2 or CO; r is selected from C1-C4 alkyl, C1-C4 alkoxy, halogen, halogen substituted C1-C4 alkyl, halogen substituted C1-C4 alkoxy and phenyl, and naphthyl is formed by the R and the connected phenyl; the number of R can be one or more. The synthesis method of the compound is simple and convenient, the compound is suitable for industrial production, and biological activity tests show that the compound has anti-tumor activity and can be applied to preparation of anti-tumor drugs. # imgabs1 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and specifically relates to 1-aryl-3-{4-{1-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea compounds and preparation methods thereof and applications thereof in the preparation of anti-tumor drugs. Background Art

[0002] Cancer refers to malignant tumor cells formed by genetic abnormalities in normal cells due to the action of various carcinogenic factors. Cancer has biological characteristics such as abnormal cell differentiation and proliferation, uncontrolled growth, invasiveness and metastasis. Its occurrence is a complex process with multiple factors and steps.

[0003] The development of cancer treatment strategies is a highly concerned issue today. At present, anti-tumor drugs can cure or control many types of tumors, but overall they have not yet reached a satisfactory stage. In recent years, with the development of multidisciplinary studies, people have gained a deeper understanding of the mechanisms of tumor occurrence and the mechanisms of action of anti-tumor drugs, including cytotoxic anti-tumor drugs and small molecule targeted anti-tumor drugs. However, many tumor cells will develop drug resistance during chemotherapy. Therefore, while improving the efficacy of developed drugs and reducing side effects, it is also necessary to synthesize new small molecule drugs and develop new low-toxic and effective anti-tumor drugs.

[0004] Urea compounds play an important role in the development of anti-tumor drugs because urea can form multiple stable hydrogen bonds with protein and receptor targets. This stable drug-target interaction makes urea compounds have good biological activity and drug properties. Typical anti-tumor drugs, such as sorafenib, lenvatinib and regorafenib, are all marketed drugs with diaryl urea structures (Zheng A, Chevalier N, Calderoni M, Dubuis G, Dormond O, Ziros PG, Sykiotis GP, Widmann C. CRISPR / Cas9 genome-wide screening identifies KEAP1 as a sorafenib, lenvatinib, and regorafenib sensitivity gene in hepatocellular carcinoma. Oncotarget, 2019, 10(66): 7058-7070.).

[0005] Structure of sorafenib:

[0006]

[0007] Structure of lenvatinib:

[0008]

[0009] Structure of regorafenib:

[0010] Summary of the invention

[0011] The primary purpose of the present invention is to provide a 1-aryl-3-(heterocyclic substituted phenyl) urea compound or a pharmaceutically acceptable salt thereof as shown in Formula I:

[0012]

[0013] Y is CH2 or CO;

[0014] R is selected from C1-C4 alkyl, C1-C4 alkoxy, halogen, halogen-substituted C1-C4 alkyl, halogen-substituted C1-C4 alkoxy, phenyl, and naphthyl formed with the connected phenyl; R can be 1 or more.

[0015] Furthermore, R is selected from methyl, methoxy, trifluoromethyl, trifluoromethoxy, halogen, phenyl, and naphthyl formed with the connected phenyl; R can be 1 or more.

[0016] Furthermore, R is selected from 4-chloro-3-trifluoromethyl, 4-methoxy, 4-fluoro, 3-chloro-4-fluoro, 4-methyl, 4-trifluoromethoxy, 4-chloro, 4-phenyl, 4-trifluoromethyl, 3-trifluoromethyl, 3-methyl, and the attached phenyl group to form a naphth-1-yl group.

[0017] The present invention preferably comprises the following compounds:

[0018] 1-(4-chloro-3-trifluoromethylphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0019] 1-(4-methoxyphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0020] 1-(3-Trifluoromethylphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0021] 1-(4-Fluorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0022] 1-(3-Chloro-4-fluorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0023] 1-(p-Tolyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0024] 1-(4-Trifluoromethoxyphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0025] 1-(4-Chlorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0026] 1-(Naphthalen-1-yl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0027] 1-(Biphenyl-4-yl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea;

[0028] 1-(4-Fluorophenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea;

[0029] 1-(4-Trifluoromethoxyphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea;

[0030] 1-(3-Trifluoromethylphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea;

[0031] 1-(3-Chloro-4-fluorophenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea;

[0032] 1-(3-Methylphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea.

[0033] A pharmaceutical composition comprises as an active ingredient a compound represented by formula I of the present invention, its prodrug and pharmaceutically active metabolite, and any one of its pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier or diluent.

[0034] "Pharmaceutically acceptable salts" refer to conventional acid addition salts or base addition salts which retain the biological effectiveness and properties of the compounds of Formula I and which are formed with suitable non-toxic organic or inorganic acids or organic or inorganic bases. Acid addition salts include hydrochlorides, hydrobromides, hydroiodides, nitrates, phosphates, sulfates, perchlorates, thiocyanates, bisulfates, persulfates, borates, formates, acetates, propionates, valerates, pivalates, caproates, heptanoates, caprylates, isooctanoates, undecanoates, laurates, palmitates, stearates, oleates, cyclopropionates, oxalates, malonates, succinates, maleates, fumarates, adipates, azelaates, acrylates, crotonates, tiglates, itaconates, sorbates, cinnamates, glycolates, lactates, malates, tartrates, citrates, tartrates, mandelates, benzilates, tropicates, ascorbic acids, gluconates, glucoheptonates, glucarates, mannonates, lactobionates, benzoates, phthalates, terephthalates, furoates, nicotinates, isonicotinates, Salicylate, acetylsalicylate, butyrate, gallate, caffeate, ferulate, picrate, camphorate, camphorsulfonate, methanesulfonate, ethanesulfonate, propanesulfonate, benzenesulfonate, p-toluenesulfonate, p-aminobenzenesulfonate, sulfamate, taurine, 2-hydroxyethanesulfonate, glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartate, asparagine, glutamate, lysine, glutamine, methionine, serine, threonine, cysteine, proline, histidine, arginine, edetate, pyruvate, α-ketoglutarate, alginate, cyclopentanepropionate, 3-phenylpropionate, 3-cyclohexylpropionic acid, 2-naphthoate, 2-naphthalenesulfonate, pamoate, lauryl sulfate, glycerophosphate, lauryl sulfate, pectinate, etc. Basic salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salts of organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine salts, etc., and basic nitrogen-containing groups can be quaternized with such agents as lower alkyl halides such as methyl, ethyl, propyl and butyl chloride, bromide and iodide; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and diamyl sulfate; long chain halides such as decyl, lauryl, myristyl and stearyl chloride, bromide and iodide; aralkyl halides such as benzyl and phenethyl bromide, etc. Preferred acids for forming acid addition salts include hydrochloric acid, acetate, p-toluenesulfonic acid, methanesulfonic acid, maleic acid, malic acid, picric acid, citric acid, p-aminobenzenesulfonic acid.

[0035] The present invention also relates to a pharmaceutical composition for inhibiting tyrosine kinase and serine threonine kinase, which contains a compound or derivative represented by formula I or a pharmaceutically suitable acid addition salt thereof and a pharmaceutically acceptable carrier.

[0036] "Pharmaceutically acceptable" such as pharmaceutically acceptable carriers, excipients, prodrugs, etc., means pharmacologically acceptable and substantially non-toxic to the patient to whom the particular compound is administered.

[0037] "Pharmaceutically active metabolite" refers to a metabolic product of a compound of Formula I that is pharmaceutically acceptable and effective.

[0038] The term "halogen" as used herein includes fluorine, chlorine, bromine and iodine.

[0039] The compounds of the invention can be administered to patients by various methods, such as oral administration in capsules or tablets, administration in sterile solutions or suspensions, and in some cases, intravenous injection in the form of solutions. The free base compounds of the invention can be formulated and administered in the form of their pharmaceutically suitable acid addition salts.

[0040] The following scheme summarizes the preparation steps of the compounds of the present invention:

[0041]

[0042]

[0043] R in the synthetic route is the corresponding group at the corresponding position of the compound shown in Formula I.

[0044] The present invention also provides the use of 1-aryl-3-(heterocyclic substituted phenyl) urea compounds shown in formula I or pharmaceutically acceptable salts thereof or the above pharmaceutical compositions in the preparation of A549 cell, A375 cell, MCF-7 cell and HeLa cell proliferation inhibitors.

[0045] The present invention also provides the use of 1-aryl-3-(heterocyclic substituted phenyl) urea compounds or pharmaceutically acceptable salts thereof or the above pharmaceutical compositions shown in formula I in the preparation of anti-tumor drugs. The tumors include lung cancer, melanoma, breast cancer, human cervical cancer and ovarian cancer.

[0046] Beneficial effects of the present invention:

[0047] The present invention discloses a series of novel 1-aryl-3-(heterocyclic substituted phenyl) urea compounds, which exhibit different degrees of proliferation inhibition activity on tumor cell lines such as A549 (lung cancer cells), A375 (melanoma cells), MCF-7 (breast cancer cells) and HeLa (human cervical cancer cells) in vitro experiments, and are further applied to the research and preparation of anti-tumor drugs. The present invention provides a new strategy and potential drug for cancer treatment. The synthesis method of the compound of the present invention is simple and suitable for industrial production. DETAILED DESCRIPTION

[0048] The present invention is further described in detail below with specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments, and all technologies realized based on the content of the present invention belong to the scope of the present invention.

[0049] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents and instruments used, they are all conventional reagent products that can be purchased commercially.

[0050] Example 1: 1-(4-chloro-3-trifluoromethylphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-[5-yl]methyl]phenyl]urea (TT01)

[0051] Step a: Preparation of (3aR,6R,6aR)-6-hydroxymethyl-2,2-dimethyltetrahydrofuran[3,4-d][1,3]dioxolane-4-ol

[0052] Weigh D-ribose (10.0 g, 66.7 mmol) and place it in a flask, add acetone (50 mL), slowly drop sulfuric acid (0.5 mL) in an ice bath, stir and react at room temperature for 1.5 h, then add triethylamine (TEA) to neutralize the reaction solution to neutrality. After evaporating the acetone under reduced pressure, separate the solution by silica gel column chromatography (PE:EA=2:1) ​​to obtain 11.87 g of a light yellow viscous liquid with a yield of 93.7%.

[0053] Step b: Preparation of [(3aR,4R,6aR)-6-hydroxy-2,2-dimethyltetrahydrofuran[3,4-d][1,3]dioxolan-4-yl]methyl p-toluenesulfonate

[0054] Weigh (3aR, 6R, 6aR)-6-hydroxymethyl-2,2-dimethyltetrahydrofuran [3,4-d] [1,3] dioxolane-4-ol (11.87 g, 62.4 mmol) into a flask, add 20 mL of pyridine to dissolve, weigh p-toluenesulfonyl chloride (TsCl, 14.28 g, 74.9 mmol) and dissolve in 50 mL of pyridine (Py), slowly add to the reaction system under ice bath, stir and react overnight at room temperature after addition. Dilute the reaction solution with water, extract with ethyl acetate, combine the organic phases, wash with saturated brine, dry over anhydrous sodium sulfate, evaporate most of the ethyl acetate under reduced pressure, and separate by column chromatography (PE: EA = 5: 1) to obtain 14.00 g of white solid, with a yield of 67.3%.

[0055] Step c: (3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation

[0056] [(3aR,4R,6aR)-6-hydroxy-2,2-dimethyltetrahydrofuran[3,4-d][1,3]dioxolan-4-yl]methyl p-toluenesulfonate (0.35 g, 1.0 mmol), methylhydrazine sulfate (0.17 g, 1.2 mmol), acetonitrile (2 mL), triethylamine (0.55 mL, 4.0 mmol) were added to a flask and stirred at 35°C under nitrogen protection for overnight reaction. Most of the acetonitrile was evaporated under reduced pressure and then separated by column chromatography (PE:EA=2:1) ​​to obtain 0.133 g of a slightly yellow waxy solid with a yield of 66.3%. 1 H NMR (400MHz, CDCl3) δ4.88–4.86(m,1H),4.86–4.81(m,1H),4.80–4.73(m,1H),4.27(m,J=2.1,0. 9Hz,1H),2.75–2.71(m,1H),2.36(dd,J=11.3,2.0Hz,1H),2.34(s,3H),1.47(s,3H),1.35(s,3H). 13 C NMR (100MHz, CDCl3) δ111.42,91.50,82.78,82.73,78.76,59.98,46.24,26.02,24.54.MS:201.1([M+H] + ).

[0057] Step d: (3aR,4R,8R,8aR)-2,2,6-trimethyl-5-(4-nitrobenzyl)hexahydro-4H-4,8-epoxy[1,3]dioxolano[4,5-d][1,2]diazepine Preparation

[0058] Weigh (3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine (2g, 10mmol), p-nitrobenzyl bromide (2.6g, 12mmol), potassium carbonate (6.0g, 45mmol) were placed in a 100mL round-bottom flask, a stirrer was added, 35mL of acetonitrile was added, a drying tube was added, the temperature was set to 60°C, and condensation reflux was performed for 48h. After the reaction was completed, it was extracted three times with 50mL of ethyl acetate, washed three times with 50mL of water, washed three times with 50mL of saturated brine, dried, most of the ethyl acetate was evaporated under reduced pressure, and then separated by column chromatography (PE:EA=6:1) to obtain 2.5g of yellow solid, with a yield of 74.5%; Mp: 190.3-192.5°C; MS: 336.1 ([M+H] + ).

[0059] Step e: 4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-amino-5-yl]methyl}aniline

[0060] Weigh (3aR,4R,8R,8aR)-2,2,6-trimethyl-5-(4-nitrobenzyl)hexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine (3.35g, 10mmol), ferric chloride hexahydrate (0.27g, 1mmol), and activated carbon 1g were placed in a 100mL round-bottom flask, a stirrer was added, 25mL of anhydrous ethanol was added, and the temperature was set to 80°C. Under heating and stirring conditions, hydrazine hydrate (3.03mL, 50mmol) was added dropwise, condensed and refluxed, and the reaction was carried out for 12h. After most of the ethanol was evaporated under reduced pressure, 1.25g of yellow solid was obtained by column chromatography (PE:EA=3:1), with a yield of 40.9%; Mp:161.1-163.2℃; MS:328.1([M+Na] + ).

[0061] Step f: 1-(4-chloro-3-trifluoromethylphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-[5-yl]methyl]phenyl]urea (TT01)

[0062] Weigh triphosgene (BTC) (0.2 g, 0.66 mmol) and dichloromethane (DCM, 10 mL) into a flask, slowly drop a dichloromethane solution of 4-chloro-3-trifluoromethylaniline (0.39 g, 2 mmol), then slowly drop a dichloromethane solution of triethylamine (0.4 g, 4 mmol), then slowly drop a dichloromethane solution of 4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}aniline (0.31 g, 1 mmol) in dichloromethane solution, stirred and reacted at room temperature overnight. After most of the dichloromethane was evaporated under reduced pressure, 0.48 g of white solid was obtained by column chromatography (DCM:EA=15:1), with a yield of 90.6%; Mp: 120.1-123.3°C; 1 H NMR (400MHz, DMSO-d6) δ9.12 (s, 1H), 8.81 (s, 1H), 8.11 (d, J = 2.4Hz, 1H), 7.63 (dd, J = 8 .8,2.5Hz,1H),7.58(d,J=8.8Hz,1H),7.42(d,J=8.3Hz,2H),7.23(d,J=8.2Hz,2H),4. 67(s,2H),4.28–4.23(m,1H),4.19–4.14(m,1H),3.92–3.78(m,2H),2.99(dd,J=11.8, 2.1Hz,1H),2.39(s,3H),2.37–2.32(m,1H),1.27(s,3H),1.22(s,3H); MS:527.1([M+H] + ).

[0063] Example 2: 1-(4-methoxyphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-[5-yl]methyl]phenyl]urea (TT02)

[0064] Referring to the preparation method of Example 1, 4-chloro-3-trifluoromethylaniline in step f was replaced by 4-methoxyaniline to obtain 0.23 g of a white solid with a yield of 50.6%; Mp: 174.7-186.4°C; 1H NMR (400MHz, DMSO-d6) δ8.56 (s, 1H), 8.44 (s, 1H), 7.39 (d, J = 8.5Hz, 2H), 7.35 (d,J=9.0Hz,2H),7.20(d,J=8.4Hz,2H),6.86(d,J=9.0Hz,2H),4.67(s,2H),4. 30–4.21(m,1H),4.19–4.14(m,1H),3.93–3.78(m,2H),3.71(s,3H),3.00(dd, J=11.7,2.1Hz,1H),2.40(s,3H),2.38–2.34(m,1H),1.28(s,3H),1.23(s,3H). 13 C NMR(101MHz,DMSO-d6)δ154.90,153.16,139.25,131.21,130.17,120.43,118.50,114.44,11 0.81,92.79,83.37,82.69,79.05,55.63,49.93,42.43,42.19,26.43,24.83; MS:455.0([M+H] + ).

[0065] Example 3: 1-(3-trifluoromethylphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-methyl]phenyl]urea (TT03)

[0066] Refer to the preparation method of Example 1, and replace 4-chloro-3-trifluoromethylaniline in step f with 3-trifluoromethylaniline to obtain 0.34 g of white solid, with a yield of 69.0%; Mp: 169.0-171.6°C; 1H NMR(400MHz,DMSO-d6)δ9.01(s,1H),8.77(s,1H),8.02(s,1H),7.56(d,J=9.1Hz,1H),7 .53–7.47(m,1H),7.43(d,J=8.4Hz,2H),7.30(d,J=7.5Hz,1H),7.23(d,J=8.5Hz,2H),4 .67(s,2H),4.30–4.22(m,1H),4.20–4.13(m,1H),3.94–3.80(m,2H),3.00(dd,J=11.8, 2.1Hz,1H),2.40(s,3H),2.38–2.33(m,1H),1.28(s,3H),1.22(s,3H); MS:493.1([M+H] + ).

[0067] Example 4: 1-(4-fluorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-[5-yl]methyl]phenyl]urea (TT04)

[0068] Refer to the preparation method of Example 1, and replace 4-chloro-3-trifluoromethylaniline in step f with 4-fluoroaniline to obtain 0.32 g of white solid, with a yield of 72.3%; Mp: 189.4-190.7°C; 1 H NMR(400MHz, CDCl3)δ7.46(s,1H),7.30(s,1H),7.28–7.23(m,2H),7.19–7.07(m,4 H),6.95–6.84(m,2H),4.85(d,J=5.5Hz,1H),4.74(d,J=5.6Hz,1H),4.53–4.48(m, 1H),4.34–4.29(m,1H),4.01–3.92(m,1H),3.88–3.78(m,1H),3.17(dd,J=11.9,2. 1Hz,1H),2.49(s,3H),2.35–2.28(m,1H),1.39(s,3H),1.28(s,3H); MS:440.9([MH] + ).

[0069] Example 5: 1-(3-chloro-4-fluorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-[5-yl]methyl]phenyl]urea (TT05)

[0070] Refer to the preparation method of Example 1, and replace 4-chloro-3-trifluoromethylaniline in step f with 3-chloro-4-fluoroaniline to obtain 0.31 g of white solid, with a yield of 65.0%; Mp: 204.7-206.5°C; 1 H NMR (400MHz, DMSO-d6) δ8.84 (s, 1H), 8.73 (s, 1H), 7.80 (dd, J = 6.8, 2.4Hz, 1H), 7. 40(d,J=8.5Hz,2H),7.37–7.25(m,2H),7.22(d,J=8.5Hz,2H),4.77–4.58(m,2H), 4.36–4.20(m,1H),4.19–4.06(m,1H),3.97–3.75(m,2H),3.00(dd,J=11.7,2.1Hz ,1H),2.40(s,3H),2.38–2.34(m,1H),1.28(s,3H),1.23(s,3H); MS:477.0([M+H] + ).

[0071] Example 6: 1-(p-Tolyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-methyl]phenyl]urea (TT06)

[0072] Refer to the preparation method of Example 1, and replace 4-chloro-3-trifluoromethylaniline in step f with 4-methylaniline to obtain 0.37 g of white solid, with a yield of 84.4%; Mp: 200.5-202.4°C; 1 H NMR (400MHz, DMSO-d6) δ8.59(s,1H),8.51(s,1H),7.39(d,J=8.5Hz,2H),7.32(d,J= 8.4Hz,2H),7.21(d,J=8.5Hz,2H),7.07(d,J=8.2Hz,2H),4.71–4.63(m,2H),4.28–4 .23(m,1H),4.19–4.14(m,1H),3.88–3.79(m,2H),3.00(dd,J=11.7,2.1Hz,1H),2.4 0(s,3H),2.38–2.34(m,1H),2.24(s,3H),1.28(s,3H),1.23(s,3H); MS:439.0([M+H] + ).

[0073] Example 7: 1-(4-trifluoromethoxyphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 1-(5-yl]methyl}phenyl}urea (TT07) was prepared by the method of Example 1, and 4-chloro-3-trifluoromethylaniline in step f was replaced by 4-trifluoromethoxyaniline to obtain 0.43 g of a white solid with a yield of 84.6%; Mp: 175.8-176.7°C. 1 H NMR (400MHz, CDCl3) δ7.71 (s, 1H), 7.42 (s, 1H), 7.36 (d, J = 9.0Hz, 2H), 7.20–7.13 (m, 4H),7.05(d,J=9.1Hz,2H),4.86(d,J=5.6Hz,1H),4.76(d,J=5.5Hz,1H),4.54–4.49( m,1H),4.36–4.31(m,1H),3.99–3.91(m,1H),3.85–3.78(m,1H),3.19(dd,J=11.9,2. 1Hz,1H),2.50(s,3H),2.36–2.30(m,1H),1.41(s,3H),1.30(s,3H); MS:509.0([M+H] + ).

[0074] Example 8: 1-(4-chlorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-[5-yl]methyl]phenyl]urea (TT08)

[0075] Refer to the preparation method of Example 1, and replace 4-chloro-3-trifluoromethylaniline in step f with 4-chloroaniline to obtain 0.37 g of white solid, with a yield of 80.6%; Mp: 202.0-203.5°C; 1H NMR (400MHz, CDCl3) δ7.52 (s, 1H), 7.33 (s, 1H), 7.29 (d, J = 8.8Hz, 2H), 7.20–7. 13(m,6H),4.88(d,J=5.5Hz,1H),4.75(d,J=5.6Hz,1H),4.55–4.50(m,1H),4.3 5–4.30(m,1H),4.02–3.94(m,1H),3.89–3.81(m,1H),3.18(dd,J=11.9,2.1Hz, 1H),2.52(s,3H),2.37–2.30(m,1H),1.39(s,3H),1.28(s,3H); MS:459.0([M+H] + ).

[0076] Example 9: 1-(Naphthalen-1-yl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-[5-yl]methyl]phenyl]urea (TT09)

[0077] Refer to the preparation method of Example 1, and replace 4-chloro-3-trifluoromethylaniline in step f with 1-naphthylamine to obtain 0.35 g of white solid, with a yield of 73.8%; Mp: 210.1-211.6°C; 1 H NMR (400MHz, CDCl3) δ7.88(d,J=8.4Hz,1H),7.72(d,J=7.6Hz,1H),7.56(d,J=6.9Hz,2H),7.46 –7.23(m,5H),7.13(d,J=8.5Hz,2H),7.07(d,J=8.2Hz,2H),4.78(d,J=5.6Hz,1H),4.67(d,J=5. 6Hz,1H),4.50–4.39(m,1H),4.24–4.19(m,1H),3.92–3.83(m,1H),3.78–3.67(m,1H),3.07(dd, J=11.7,2.1Hz,1H),2.41(s,3H),2.25–2.19(m,1H),1.37(s,3H),1.25(s,3H); MS:475.1([M+H] + ).

[0078] Example 10: 1-(Biphenyl-4-yl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-methyl]phenyl]urea (TT10)

[0079] Refer to the preparation method of Example 1, and replace 4-chloro-3-trifluoromethylaniline in step f with 4-aminobiphenyl to obtain 0.44 g of white solid with a yield of 88.0%; Mp: 168.7-169.3°C. 1 H NMR (400MHz, CDCl3) δ7.51 (s, 1H), 7.52–7.34 (m, 8H), 7.33 (d, J = 3.2Hz, 1H), 7.30–7.24 (m, 1H), 7. 23(d,J=5.8Hz,2H),7.18(s,1H),6.57(d,J=4.7Hz,0H),4.86(d,J=5.5Hz,1H),4.74(d,J=5.6Hz,1 H),4.53(s,1H),4.32(d,J=2.1Hz,1H),3.97(d,J=11.7Hz,1H),3.83(d,J=11.7Hz,1H),3.16(dd,J =11.9,2.1Hz,1H),2.49(s,3H),2.31(d,J=11.7Hz,1H),1.40(s,3H),1.29(s,3H); MS:501.0([M+H] + ).

[0080] Example 11: 1-(4-Fluorophenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-carbonyl]phenyl}urea (TT11)

[0081] Referring to the preparation method of steps ac of Example 1, (3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine was obtained. (Compound 4 in the synthetic route).

[0082] Step g: (4-nitrophenyl) [(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy [1,3] dioxolane [4,5-d] [1,2] diazepine Preparation of 5-methyl-1-methanone

[0083] Weigh p-nitrobenzoic acid (1.67 g, 10 mmol) into a 50 mL round-bottom flask, add 15 mL of dichloromethane, add thionyl chloride (0.89 g, 7.5 mmol) under ice bath, stir at room temperature for 1 h, add pyridine (1.67 mL, 20 mmol) and (3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine (1.0 g, 5 mmol), react for 24 h. Extract three times with 20 mL of dichloromethane, wash once with saturated sodium bicarbonate, wash three times with 20 mL of water, and wash once with saturated sodium chloride. Obtain 1.2 g of white solid, yield 68.7%; Mp: 162.6-165.7 ° C; MS: 350.0 ([M+H] + ).

[0084] Step h: 4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-amino-5-yl]methyl}aniline

[0085] Weigh (4-nitrophenyl) [(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy [1,3] dioxolane [4,5-d] [1,2] diazepine -5-yl) ketone (3.35g, 10mmol), ferric chloride hexahydrate (0.056g, 0.2mmol), and activated carbon 2.8g were placed in a 50mL round-bottom flask, and 20mL of anhydrous ethanol was added. The temperature was set to 80°C. Under heating and stirring conditions, hydrazine hydrate (0.44g, 7mmol) was added dropwise, condensed and refluxed, and the reaction was carried out for 12h. After filtering and decompression to evaporate most of the ethanol, 0.4681g of yellow solid was obtained by column chromatography (PE:EA=4:1), with a yield of 52.3%; Mp:143.3-145.5℃; MS:320.2([M+H] + ).

[0086] Step i: 1-(4-fluorophenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-carbonyl]phenyl}urea (TT11)

[0087] Weigh triphosgene (0.2 g, 0.66 mmol) and dichloromethane (10 mL) into a flask, slowly drop a dichloromethane solution of p-fluoroaniline (0.22 g, 2 mmol), then slowly drop a dichloromethane solution of triethylamine (0.4 g, 4 mmol), then slowly drop 4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}aniline (0.32 g, 1 mmol) in dichloromethane solution, stirred and reacted at room temperature overnight. After most of the dichloromethane was evaporated under reduced pressure, 0.42 g of white solid was obtained by column chromatography (DCM:EA=15:1), with a yield of 92.4%; Mp: 190.0-190.9°C; 1 H NMR (400MHz, CDCl3) δ7.80 (s, 1H), 7.65 (s, 1H), 7.42 (d, J = 8.4Hz, 2H), 7.22– 7.20(m,2H),7.19–7.17(m,2H),6.93–6.83(m,2H),5.54–5.49(m,1H),5.00–4 .94(m,1H),4.80–4.73(m,1H),4.34–4.29(m,1H),3.56(dd,J=12.1,2.7Hz,1H ),2.84(s,3H),2.59–2.47(m,1H),1.36(s,3H),1.30(s,3H); MS:457.1([M+H] + ).

[0088] Example 12: 1-(4-trifluoromethoxyphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-carbonyl]phenyl}urea (TT12)

[0089] Refer to the preparation method of Example 11, and replace the p-fluoroaniline in step i with 4-trifluoromethoxyaniline to obtain 0.47 g of white solid, with a yield of 92.8%; Mp: 130.4-132.0°C; 1H NMR (400MHz, CDCl3) δ7.74 (s, 1H), 7.43 (d, J = 8.4Hz, 2H), 7.34 (d, J = 9.0Hz, 2H) ,7.21(s,1H),7.20–7.10(m,2H),7.07(d,J=8.4Hz,2H),5.58–5.48(m,1H),5.0 3–4.98(m,1H),4.83–4.78(m,1H),4.38–4.33(m,1H),3.57(dd,J=11.6,2.7Hz, 1H),2.87(s,3H),2.64–2.56(m,1H),1.37(s,3H),1.31(s,3H); MS:523.1([M+H] + ).

[0090] Example 13: 1-(3-trifluoromethylphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-carbonyl]phenyl}urea (TT13)

[0091] Refer to the preparation method of Example 11, and replace the p-fluoroaniline in step i with 3-trifluoromethylaniline to obtain 0.42 g of white solid, with a yield of 82.9%; Mp: 182.3-184.0°C; 1 H NMR (400MHz, CDCl3) δ7.97(s,1H),7.89(s,1H),7.63(s,1H),7.55(d,J=8.8Hz,1H ),7.44(d,J=8.4Hz,2H),7.34–7.26(m,1H),7.27–7.20(m,3H),5.58–5.48(m,1H) ,5.05–4.99(m,1H),4.87–4.81(m,1H),4.39–4.34(m,1H),3.60(dd,J=12.0,2.8H z,1H),2.89(s,3H),2.70–2.62(m,1H),1.37(s,3H),1.31(s,3H); MS:507.1([M+H] + ).

[0092] Example 14: 1-(3-chloro-4-fluorophenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-carbonyl]phenyl}urea (TT14)

[0093] Refer to the preparation method of Example 11, and replace the p-fluoroaniline in step i with 3-chloro-4-fluoroaniline to obtain 0.39 g of white solid, with a yield of 79.5%; Mp: 151.4-154.3°C; 1 H NMR (400MHz, CDCl3) δ7.91(s,1H),7.84(s,1H),7.42(d,J=8.6Hz,2H),7.34(dd,J= 6.5,2.7Hz,1H),7.17(d,J=8.2Hz,2H),7.11–7.01(m,1H),7.00–6.87(m,1H),5.59 –5.42(m,1H),5.01–4.93(m,1H),4.83–4.73(m,1H),4.37–4.30(m,1H),3.54(dd,2 H),2.83(s,3H),2.57(d,J=12.3Hz,1H),1.36(s,3H),1.30(s,3H);MS:491.1([M+H] + ).

[0094] Example 15: 1-(3-methylphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine Preparation of 5-carbonyl]phenyl}urea (TT15)

[0095] Refer to the preparation method of Example 11, and replace the p-fluoroaniline in step i with m-toluidine to obtain 0.31 g of a white solid with a yield of 68.3%; Mp: 198.9-202.8°C; 1 H NMR (400MHz, CDCl3) δ7.78 (s, 1H), 7.54 (s, 1H), 7.42 (d, J = 8.6Hz, 2H), 7.22 (d,J=4.7Hz,2H),7.12(d,J=8.6Hz,2H),6.98(d,J=7.8Hz,2H),5.60–5.45(m ,1H),5.00–4.93(m,1H),4.79–4.72(m,1H),4.33–4.28(m,1H),3.56(dd,2H) ,2.83(s,3H),2.56–2.48(m,1H),1.36(s,3H),1.30(s,3H); MS:453.1([M+H] + ).

[0096]

[0097]

[0098]

[0099] Example 16: Pharmacological Example

[0100] Experimental methods:

[0101] a. Cell recovery

[0102] The cryopreserved tubes containing melanoma cells A375, non-small cell lung cancer cells A549, human cervical cancer cells HeLa, and human breast cancer cells MCF-7 were taken out of the liquid nitrogen tank, shaken in a 37°C water bath to dissolve, and the dissolved cryopreserved solution was transferred to a 15mL centrifuge tube containing culture medium, centrifuged at 1050rpm for 3min, the supernatant was discarded, 1mL of complete culture medium was added to resuspend the cells, and inoculated into a 75mL culture bottle pre-filled with warm complete culture medium. After shaking the culture bottle to evenly distribute the cells, it was placed in an incubator at 37°C and 5% CO2 for culture, and the cell morphology and growth were observed at regular intervals.

[0103] b. Cell passaging

[0104] When the cells in the culture flask grow to 80%-90%, discard the culture medium, add 3mL 0.25% trypsin solution and digest for 2min in a 37℃ incubator, add 6mL culture medium to terminate digestion, blow the flask wall repeatedly, transfer the detached cells to a 15mL centrifuge tube, centrifuge at 1050rpm for 3min, discard the supernatant, add complete culture medium containing 10% fetal bovine serum to resuspend the cells, make a single cell suspension, blow evenly, and count. Press 100μL 2-5*10 3 The cells were inoculated into 96-well culture plates, and the edge wells were filled with sterile PBS to avoid edge effects. The plates were placed in an incubator at 37°C and 5% CO2 for 24 hours.

[0105] c. Cytotoxicity and proliferation inhibition

[0106] After the cells in the above 96-well culture plate were cultured in an incubator for 24 hours, the culture medium was discarded and replaced with 200 μL of complete culture medium containing the sample for a further 72 hours. 3-5 concentration gradients were set for each group of samples, and 3 replicate wells were set for each concentration. 20 μL of MTT solution (5 mg / mL, prepared with PBS, pH = 7.4) was added to each well and incubated for another 4 hours. 150 μL of DMSO was added to each well and shaken for 3 minutes to fully dissolve the crystals. The light absorption value of each well was measured on an ELISA reader at 490 nm, 570 nm or 630 nm, and the results were recorded. The cellular drug response of the sample (the concentration required to obtain 50% maximum cellular effect, EC 50 ) was obtained by calculating the experimental data using the software GraphPad Prism8.0.1.

[0107] Experimental results:

[0108]

[0109] Among the 15 compounds mentioned above, all compounds showed different degrees of tumor cell proliferation inhibition activity against A549, MCF-7, A375 and HeLa cell lines. Among them, TT01 had significant proliferation inhibition activity against A549 cell line, and TT02 and TT14 had significant proliferation inhibition activity against HeLa cell line. 50 All were below 10μM.

[0110] Formulation Examples

[0111] The following formulation examples are merely illustrative of the scope of protection of the present invention, but are not intended to limit the scope of protection of the present invention in any way. The active compounds described in the following examples are compounds TT01-TT15 prepared in the above examples.

[0112] Example 17: Tablet Formulation

[0113] Active compound 25-1000 mg, starch 45 mg, microcrystalline cellulose 35 mg, polyvinyl pyrrolidone (as a 10% aqueous solution) 4 mL, sodium carboxymethyl cellulose 4.5 mg, magnesium stearate 0.5 mg, talc 1 mg.

[0114] Example 18: Suspension Concentrate Formulation

[0115] Active compound 0.1-1000 mg, sodium carboxymethyl cellulose 50 mg, syrup 1.25 mg, sodium benzoate 0.1 mg, flavoring agent 25 mg, coloring agent 5 mg, add purified water to 5 mL.

[0116] Example 19: Aerosol Formulation

[0117] Active compound 0.25 mg, ethanol 25-75 mL, propellant 22 (chlorodifluoromethane) 70 mg.

[0118] Example 20: Suppository Formulation

[0119] Active compound 250mg, saturated fatty acid glycerides 2000mL.

[0120] Example 21: Injectable Formulation

[0121] Active compound 50 mg, isotonic saline solution 1000 mL.

[0122] Example 22: Ointment Formulation

[0123] 0.025 g of micronized active compound, 10 g of liquid paraffin, and add soft white wax to 100 g.

[0124] Example 23: Ointment Formulation

[0125] 0.025 g of active compound, 5 g of propylene glycol, 5 g of sorbitan sesquioleate, 10 g of liquid paraffin, and add soft white wax to 100 g.

[0126] Example 24: Oil-in-water cream formulation

[0127] Active compound 0.025g, hexadecanol 5g, glyceryl monostearate 5g, liquid paraffin 10g, cetyl alcohol polyoxyethylene ether 2g, citric acid 0.1g, sodium citrate 0.2g, propylene glycol 35g, add water to 100g.

[0128] Example 25: Oil-in-water cream formulation

[0129] 0.025 g of micronized active compound, 15 g of soft white wax, 5 g of liquid paraffin, 5 g of hexadecanol, 2 g of Sorbimacrogolstearate (Tween 65 of specific pharmaceutical excipient grade), 0.5 g of sorbitan monostearate, 0.2 g of sorbic acid, 0.1 g of citric acid, 0.2 g of sodium citrate, add water to 100 g.

[0130] Example 26: Water-in-oil cream formulation

[0131] 0.025g active compound, 35g soft white wax, 5g liquid paraffin, 5g dehydrated sorbitan sesquioleate, 0.2g sorbic acid, 0.1g citric acid, 0.2g sodium citrate, add water to 100g.

[0132] Example 27: Lotion Formulation

[0133] Active compound 0.25 g, isopropanol 0.5 mL, carboxyvinyl polymer 3 mg, NaOH 2 mg, add water to 1 g.

[0134] Example 28: Formulation of suspension for injection

[0135] Active compound 10 mg, sodium carboxymethyl cellulose 7 mg, NaCl 7 mg, polyoxyethylene (20) sorbitan monooleate 0.5 mg, benzyl alcohol 8 mg, add sterile water to 1 mL.

[0136] Example 29: Aerosol Formulation for Oral and Nasal Inhalation

[0137] Active compound 0.1% w / w, Sorbitan trioleate 0.7% w / w, Trichlorofluoromethane 24.8% w / w, Dichlorotetrafluoroethane 24.8% w / w, Dichlorodifluoromethane 49.6% w / w.

[0138] Example 30: Nebulized solution formulation

[0139] Active compound 7 mg, propylene glycol 5 mg, add water to 10 g.

[0140] Example 31: Powder Formulation for Inhalation

[0141] Fill a clear gelatin capsule with the mixture of the following ingredients, 0.1 mg of micronized active compound, 20 mg of lactose, and inhale the powder with the aid of an inhalation device.

[0142] Example 32: Powder Formulation for Inhalation

[0143] The spheronized powder is filled into a multiple-dose powder inhaler, each dose containing 0.1 mg of the micronized active compound.

[0144] Example 33: Powder formulation for inhalation

[0145] The spheronized powder is filled into a multiple-dose powder inhaler, each dose containing 0.1 mg of micronized active compound and 1 mg of micronized lactose.

[0146] Example 34: Capsule Formulation

[0147] Active compound 1.0 mg, small sugar spheres 321 mg, Aquacoat ECD 30 6.6 mg, acetyl tributyl citrate 0.5 mg, Tween-80 0.1 mg, Eudragit L 100-55 17.5 mg, triethyl citrate 1.8 mg, talc 8.8 mg, defoaming agent MMS 0.1 mg.

[0148] Example 35: Capsule Seedling Formulation

[0149] Active compound 2.0 mg, small sugar spheres 305 mg, Aquocoat ECD 30 5.0 mg, acetyl tributyl citrate 0.4 mg, Tween-80 0.14 mg, Eudragit NE30D 12.6 mg, Eudragit S100 12.6 mg, talc 0.16 mg.

[0150] Example 36: Enema Formulation

[0151] Active compound 2 mg, sodium carboxymethyl cellulose 25 mg, disodium edetate 0.5 mg, methyl parahydroxybenzoate 0.8 mg, propyl parahydroxybenzoate 0.2 mg, sodium chloride 7 mg, citric acid 1.8 mg, Tween-80 0.01 mg, add purified water to 1 mL.

[0152] Example 37: Formulation containing liposomes

[0153] A. Preparation of Instillation Formulation

[0154] Dipalmitoyl phosphatidylcholine (45 mg), dimyristoyl phosphatidylcholine (7 mg), dipalmitoyl phosphatidylglycerol (1 mg) and active compound (5 mg) were placed in a glass tube, all components were dissolved in chloroform, most of the solvent was evaporated with N2, and then the pressure was reduced, thereby forming a lipid film on the surface of the glass tube. An aqueous solution (0.9% NaCl) was added to the lipid, and liposomes were formed at a temperature higher than the phase transition temperature of the lipid. The resulting suspension contained liposomes ranging in size from extremely small vesicles to 2 μm.

[0155] B. Preparation of Formulations for Inhalation

[0156] Liposomes were prepared according to Example A, wherein the aqueous solution contained 10% lactose, and the ratio of lactose to lipid was 7:3. The liposome suspension was frozen with dry ice and freeze-dried, and the dried product was micronized to obtain particles with a mass mean aerodynamic diameter (MMAD) of about 2 μm.

[0157] The above is only a preferred embodiment of the present invention, and does not limit the present invention in other forms. Any person skilled in the art may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. Any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention without departing from the technical solution of the present invention still belongs to the protection scope of the technical solution of the present invention.

Claims

1. A 1-aryl-3-(heterocyclic substituted phenyl) urea compound as shown in formula I or a pharmaceutically acceptable salt thereof, characterized in that: The structure of formula I is: Y is CH2 or CO; R is selected from C1-C4 alkyl, C1-C4 alkoxy, halogen, halogen-substituted C1-C4 alkyl, halogen-substituted C1-C4 alkoxy, phenyl, and naphthyl formed with the connected phenyl; R can be 1 or more.

2. The 1-aryl-3-(heterocyclic substituted phenyl) urea compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: In formula I, R is selected from methyl, methoxy, trifluoromethyl, trifluoromethoxy, halogen, phenyl, and naphthyl formed with the connected phenyl; R can be 1 or more.

3. The 1-aryl-3-(heterocyclic substituted phenyl) urea compound or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: In formula I, R is selected from 4-chloro-3-trifluoromethyl, 4-methoxy, 4-fluoro, 3-chloro-4-fluoro, 4-methyl, 4-trifluoromethoxy, 4-chloro, 4-phenyl, 4-trifluoromethyl, 3-trifluoromethyl, 3-methyl, and the connected phenyl group forms a naphth-1-yl group.

4. The 1-aryl-3-(heterocyclic substituted phenyl) urea compound or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: The 1-aryl-3-(heterocyclic substituted phenyl) urea compound is any one of the following compounds: 1-(4-chloro-3-trifluoromethylphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(4-methoxyphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(3-Trifluoromethylphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(4-Fluorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(3-Chloro-4-fluorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(p-Tolyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(4-Trifluoromethoxyphenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(4-Chlorophenyl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(Naphthalen-1-yl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(Biphenyl-4-yl)-3-{4-{[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-5H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-yl]methyl}phenyl}urea; 1-(4-Fluorophenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea; 1-(4-Trifluoromethoxyphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea; 1-(3-Trifluoromethylphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea; 1-(3-Chloro-4-fluorophenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea; 1-(3-Methylphenyl)-3-{4-[(3aR,4R,8R,8aR)-2,2,6-trimethylhexahydro-4H-4,8-epoxy[1,3]dioxolane[4,5-d][1,2]diazepine -5-carbonyl]phenyl}urea.

5. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises as an active ingredient any one of the 1-aryl-3-(heterocyclic substituted phenyl) urea compounds or pharmaceutically acceptable salts thereof according to any one of claims 1 to 4 and a pharmaceutically acceptable carrier or diluent.

6. Use of the 1-aryl-3-(heterocyclic substituted phenyl) urea compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 in the preparation of an anti-tumor drug.

7. Use of the pharmaceutical composition according to claim 5 in the preparation of anti-tumor drugs.

8. The use according to claim 6 or 7, characterized in that: The tumors include lung cancer, melanoma, breast cancer, cervical cancer and ovarian cancer.

9. The method for preparing the 1-aryl-3-(heterocyclic substituted phenyl) urea compound or a pharmaceutically acceptable salt thereof according to claim 4, characterized in that: The following steps are involved: R in the synthetic route is the corresponding group at the corresponding position of the compound described in claim 4.

Images