CN101899051B - 1-azaxanthone-3-formamide compounds as well as preparation method and antitumor application thereof - Google Patents

Abstract

The invention discloses 1-azaxanthone-3-formamide compounds as well as a preparation method and an application thereof, relating to the field of medicine. The compounds have certain antitumor effect. In particular, the compounds show good selective toxicity toward ambulant breast cancer cells, and provide a basis for developing an antitumor drug with high effect, low toxicity and strong specificity.

Description

1-Azaxanthone-3-carboxamide compounds, preparation method and anti-tumor application

technical field

The present invention relates to the technical field of medicine, more specifically, the present invention relates to a class of 1-azaxanthone-3-carboxamide compounds, and the present invention also relates to the preparation method of such compounds and their use in the preparation of antineoplastic drugs Applications. the

Background technique

Malignant tumors are one of the major diseases that seriously threaten human life and quality of life. According to data released by the Ministry of Health at the end of 2007, malignant tumors have become the number one cause of death among urban and rural residents in my country. In recent years, cancer chemotherapy has made considerable progress, and the survival time of cancer patients has been significantly prolonged. In particular, great breakthroughs have been made in the treatment of leukemia and malignant lymphoma. The treatment of more than % of solid tumors (including lung cancer, liver cancer, bladder cancer and colon cancer, etc.) still fails to achieve satisfactory results. the

With the rapid development of life science research, the tumor mechanism has been gradually elucidated, and new directions such as inhibiting tumor growth factors, interfering with tumor signal transduction, inhibiting tumor angiogenesis, and inducing tumor cell apoptosis have appeared one after another; however, due to the pathogenesis of malignant tumors It is very complicated. Clinical practice in recent years has shown that they can only achieve better clinical efficacy when they are used in combination with cytotoxic drugs. Therefore, it is of great significance to use key enzymes related to differentiation and proliferation in tumor cells as drug screening targets to discover new anticancer drugs that selectively act on specific targets with high efficiency, low toxicity and strong specificity. the

In recent years, topoisomerase I (Top1) has become one of the key target enzymes for designing new anticancer drugs. It is an essential enzyme for cell survival and participates in the whole process of DNA replication, transcription, recombination, and repair; and the content of Top1 in various tumor cells, especially lung cancer, gastric cancer, colon cancer, and ovarian cancer, is significantly higher than that in normal cells. All of these make Top1 inhibitor drugs not only have high curative effect and broad anti-tumor spectrum, but also have good selectivity for tumor cells. It has been listed as one of the six major anti-tumor drugs for key research by the National Cancer Institute of the United States. . the

Among various Top1 inhibitors, camptothecin (CPT) derivatives are the most deeply studied and the most classic Top1-specific inhibitors. Among these compounds, inotecan (CPT-11) and topotecan have been successfully marketed, and have been used for the treatment of metastatic colorectal cancer and refractory ovarian cancer, etc., and achieved good curative effect. However, this class of compounds also has the following outstanding problems: 1) In vivo metabolism is unstable, and the E-ring lactone structure necessary for activity is hydrolyzed into carboxylate form too quickly in the human body, and carboxylate form is not only ineffective for Top1, but also more easily Binding to human serum albumin; 2) The Top1 cleavage complex (Top1cc) needs to be maintained for a long time before it can be transformed into DNA damage, but camptothecin is easy to dissociate from Top1cc, so it must be prolonged when using camptothecin drugs clinically. Intravenous infusion time; 3) Poor water solubility, camptothecin unique pentacyclic conjugated planar structure has strong hydrophobicity, resulting in poor water solubility; 4) Camptothecin drugs have certain toxic and side effects, such as self-cell reduction, nausea, vomiting, etc., which limit the safe dose and thus limit the efficacy of the drug; 5) Drug resistance, there have been several reports of camptothecin-resistant mutants of Top1, the most common mutations are Asn722, Arg364, etc. Can lead to the development of camptothecin resistance. the

Non-camptothecin Top1 inhibitors have become a research hotspot of anticancer drugs in recent years. Indolocarbazole compounds are a class of in-depth research at present. J-107088 (Edotecarin) in this class of compounds has entered clinical research. However, studies have shown that this class of compounds is not a Top1-specific inhibitor, and also inhibits protein kinase C Or the activity of the checkpoint kinase Chk-1. In addition, LuotonineA and LamellarinD are also Top1 inhibitors isolated from nature in recent years, but most of them have problems such as complex structure, poor specificity, and high toxicity and side effects. Currently, only camptothecin antineoplastic drugs are widely used clinically. the

In order to discover new structural types of Top1 inhibitors, we carried out a virtual high-throughput screening study of Top1 inhibitors in the early stage, and successfully discovered several lead compounds with good drug-like properties and no potential carcinogenicity. Based on the activity, novelty and molecular graphics investigation, we finally selected and purchased some compounds for enzyme inhibitory and cytotoxic activity tests. It strongly inhibits the activity of Top1, which is significantly better than the marketed drug topotecan. The results of cytotoxic activity test show that it has certain anti-tumor activity against several tumor cells. Molecular docking studies show that this type of lead compound can form two hydrogen bonds with the key active site residue Arg364 of the Topo I target enzyme that we identified earlier, and the amide group on the side chain can interact with the DNA cleavage site on the target enzyme The cytosine at the downstream +1 position forms a selective effect, which makes this type of compound have greater advantages compared with existing Top1 inhibitors, and is expected to well overcome the problem of drug resistance of existing antitumor drugs. the

It should be noted that although individual compounds of this type are already available for purchase, the literature does not disclose the physical and chemical properties and preparation methods of such compounds, and there is no report that such compounds can be used for antitumor. Therefore, the 1-azoxanthone-3-carboxamides that we discovered for the first time represent a new class of antitumor active compounds. the

Contents of the invention

The object of the present invention is to provide a class of 1-azoxanthone-3-carboxamide compounds, or pharmaceutically acceptable salts thereof. The invention also discloses the preparation method, medical application and composition of the compound. the

The present invention further designs and synthesizes derivatives of this type of compound according to the structure of the lead obtained by our previous virtual high-throughput screening. The activity tests on various tumor cells show that this type of compound has certain anti-tumor activity, especially worth It should be pointed out that this type of compound shows good selective toxicity to metastatic breast cancer cells, which lays the foundation for the development of highly efficient, low toxicity, and strong specific antitumor drugs. the

The structure of the 1-azoxanthone-3-carboxamide compound of the present invention is shown in the general formula (I):

Or a pharmaceutically acceptable salt thereof, wherein, the substitution position of R ₁ can be located at 6 to 9 positions, and can be mono-, double-, or multi-substituted, and R ₁ is any one of the following groups: a) hydrogen; b) C1-8 linear or branched alkoxy; c) C1-8 linear or branched alkyl; d) halogen; m) methylenedioxy;

R ₂ and R ₃ independently represent the following groups: hydrogen, C1-8 linear or branched alkyl, substituted or unsubstituted aryl methylene, and the substitution is hydrogen, halogen, C1-8 straight-chain or branched-chain alkyl, C1-8 straight-chain or branched-chain alkoxy, nitro, amino, hydroxyl; or R ₂ , R ₃ together form a chain composed of 3 to 7 CH ₂ unit connections chain;

R ₄ is hydrogen, C1-8 linear or branched alkyl.

In a further preferred embodiment of the present invention, in the above-mentioned 1-azoxanthone-3-carboxamide compounds, when R ₁ is monosubstituted and the 7-position is fluorine or chlorine, the combination of R ₂ and R ₃ cannot Diethyl, tert-butyl, benzyl, p-chlorobenzyl; when R ₁ is monosubstituted and the 7th position is hydrogen, the combination of R ₂ and R ₃ cannot be benzyl, p-chlorobenzyl; when R ₁ When it is monosubstituted and the 7th position is a methyl group, the combination of _R2 and _R3 cannot be tert-butyl, benzyl, or p-chlorobenzyl.

Furthermore, in the above-mentioned 1-azoxanthone-3-carboxamide compounds, the substitution of _R1 is monosubstituted, preferably 7-position monosubstituted.

In a preferred embodiment of the present invention, in the above-mentioned 1-azoxanthone-3-carboxamide compound, the combination of R ₂ and R ₃ is a substituted or unsubstituted aryl methylene group, and the substituted It is hydrogen, halogen, C1-8 linear or branched alkyl, C1-8 linear or branched alkoxy, nitro, amino, hydroxyl; or R ₂ and R ₃ together form 3 to A chain of 7 _CH2 units connected. The aryl methylene is preferably benzyl, and the substitution on the aryl methylene is preferably hydrogen or halogen.

In a preferred embodiment of the present invention, in the 1-azoxanthone-3-carboxamide compound, R ₄ is preferably methyl.

The aryl group described in the present invention refers to the remaining atomic group after removing a hydrogen atom in an aromatic hydrocarbon molecule, which is called an aryl group. Aromatic hydrocarbons are a class of aromatic compounds, which are chemically stable, difficult to add, difficult to oxidize, easy to replace and abnormally stable in carbon rings, which are different from the properties of general saturated compounds, including: 1) single-ring aromatic hydrocarbons, The molecule contains only one benzene ring, such as benzene, toluene, ethylbenzene, styrene, etc.; 2) polycyclic aromatic hydrocarbons, the molecule contains 2 or more benzene rings, such as biphenyl, naphthalene, anthracene, phenanthrene, etc.; 3) Non-benzene aromatic hydrocarbons, which do not contain benzene rings in the molecule, but contain aromatic hydrocarbons similar in structure and properties to benzene rings, and have the characteristics of aromatic compounds, such as cyclopentadiene anion. the

The substituted aryl refers to a group obtained by replacing the hydrogen atoms on the aryl with other substituents. the

In an embodiment of the present invention, the 1-azoxanthone-3-carboxamide compound, the combination of each substituent group includes but is not limited to the following table:

Another object of the present invention is to provide the preparation method of above-mentioned 1-azoxanthone-3-carboxamides, comprising the following steps:

(1) Preparation of 3-cyano-4-benzopyrone (Ⅱ)

First react DMF and phosphorus oxychloride at 0°C for half an hour, then add substituted hydroxyacetophenone (I), stir at room temperature for 4 hours, then add hydroxylamine hydrochloride to continue the reaction for 6 hours to generate 3-cyano-4-benzo Pyrone (II);

(2) Preparation of ethyl 5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxylate (Ⅲ)

Reflux 3-cyano-4-benzopyrone (II), ethyl acetoacetate and piperidine in ethanol to generate 5-oxo-5H-[1]benzopyran[2,3-b] Ethyl pyridine-3-carboxylate (Ⅲ);

(3) Preparation of 5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxylic acid (Ⅳ)

Reflux ethyl 5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxylate (Ⅲ) in 50% H ₂ SO4-AcOH mixed acid to generate 2-methyl -5-Oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-carboxylic acid (IV);

(4) Preparation of 5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-carboxamide (Ⅴ)

Dissolve 5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxylic acid (IV) in DMF, add DCC and HOBt in dichloromethane at 0°C, After stirring for several minutes, the amine was added, and the reaction was stirred at room temperature for 24 hours to generate 5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxamide (Ⅴ). the

Certain 1-azoxanthone-3-carboxamide compounds of the present invention can be prepared in the form of their pharmaceutically acceptable salts according to conventional methods. Including its inorganic acid salts and organic acid salts: inorganic acids include (but not limited to) hydrochloric acid, sulfuric acid, phosphoric acid, diphosphoric acid, hydrobromic acid, nitric acid, etc.; organic acids include (but not limited to) acetic acid, maleic acid, fumaric acid acid, tartaric acid, succinic acid, lactic acid, p-toluenesulfonic acid, salicylic acid, oxalic acid, etc. the

The compounds of the present invention have good antitumor activity, and they can be used to treat tumors, including esophagus, stomach, intestine, rectum, oral cavity, pharynx, larynx, lung, colon, breast, uterus, endometrium, ovary, prostate, testis , bladder, kidney, liver, pancreas, bone, connective tissue, skin, eye, brain, and central nervous system, as well as thyroid cancer, leukemia, Hodgkin's disease, lymphoma, and myeloma. Preferably, the tumor to be treated is breast cancer, more preferably metastatic breast cancer. the

The pharmacological activity of the 1-azaxanthone-3-carboxamide compounds of the present invention makes it possible to be used in the preparation of antitumor, antifungal and antiviral drugs, so the present invention also includes the use of these compounds or their pharmaceutically acceptable Salt as the pharmaceutical composition of the active ingredient, the pharmaceutical composition also contains a pharmaceutically acceptable carrier, which can be in solid form or liquid form, and the pharmaceutical dosage form can be tablet, capsule, powder, granule, mixed Suspension, or injection. the

Detailed ways

The present invention will be specifically described below in conjunction with examples, but the following examples should not be regarded as limiting the scope of the present invention. the

Embodiment 1: Preparation of 3-cyano-4-benzopyrone (II)

DMF (20 mL) and POCl ₃ (12 mL, 0125 mol) were added to a round bottom flask, and the reaction was stirred at 0°C for 0.5 h. o-Hydroxyacetophenone (4.25 g, 0.031 mol) was added at 0° C., and reacted at room temperature for 4 h. After the reaction, dilute with 45 mL of CH ₂ Cl ₂ , cool to 0°C, then add a DMF solution of hydroxylamine hydrochloride (6.5 g of H ₂ NOH·HCl dissolved in 20 mL of DMF), and stir at room temperature for 24 h. After the reaction was completed, it was diluted with 35 mL of water, extracted with dichloromethane (3×25 mL), and the organic layer was washed successively with 10% NaHCO ₃ solution and saturated sodium chloride solution. Dry over anhydrous Na ₂ SO ₄ , filter, and evaporate the filtrate to give a yellow solid. Methanol was recrystallized to obtain 3.2 g of colorless needle crystals, yield 60.4.

¹ H-NMR (300MHz, CDCl ₃ ) δ: 7.53 (1H, t, 6-H), 7.55 (1H, d, 8-H), 7.78 (1H, t, 7-H), 8.25 (1H, d , 5-H), 8.40 (1H, s, 2-H); ESI-MS (m/z): 210 (M ⁺ +H+Na); IR (KBr) cm ^-1 : 2241, 1662; mp176- 178°C (lit. 177-178°C).

Example 2: Preparation of ethyl 2-methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxylate (Ⅲ)

Take 3-cyano-4-benzopyrone (II) (3.42g, 0.02mol), ethyl acetoacetate (2.60g, 0.02mol) and 4mL of piperidine, and reflux in 80mL of ethanol. Stop reflux after the reaction, cool down, add 250 mL of water to dilute, filter the precipitated solid, wash with water until neutral, and drain to obtain a red crude product. Purify on a silica gel column (eluent: petroleum ether: ethyl acetate = 20:1) to obtain 2.65 g of white crystals with a yield of 46.8%. the

¹ H-NMR (300MHz, CDCl ₃ ) δ: 1.45 (3H, t, 2'-CH ₃ ), 2.91 (3H, s, 2-CH ₃ ), 4.42 (2H, q, 1'-CH ₂ -) , 7.52(1H, t, 7-H), 7.65(1H, d, 9-H), 7.85(1H, t, 8-H), 8.28(1H, d, 6-H), 9.16(1H, s , 4-H). ESI-MS (m/z): 282 (M ⁺ +Na-K); IR (KBr) cm ^-1 : 1715, 1655; mp151-152℃

Example 3: Preparation of 2-methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxylic acid (Ⅳ)

Add ethyl 2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-carboxylate (Ⅲ) (1.5 g, 0.0053 mol) into 50% H ₂ SO ₄ -AcOH (1:2) was mixed with 27 mL of acid, and stirred under reflux at 130° C. for 3 h. The reaction solution was poured into 200 mL of water, and a solid was precipitated, filtered, washed with water, and dried to obtain an off-white solid, which was recrystallized with DMF-H ₂ O to obtain 1.21 g of a white solid, with a yield of 91.3%. ¹ H-NMR (300 MHz, d ₆ -DMSO) δ: 2.85 (3H, s, 2-CH ₃ ), 7.51 (1H, t, 7-H), 7.71 (1H, d, 9-H), 7.90 ( 1H, t, 8-H), 8.15 (1H, d, 6-H), 8.90 (1H, s, 4-H), 13.56 (1H, br, -COOH). ESI-MS (m/z): 256 (M+H ⁺ ); IR (KBr) cm ^-1 : 3103, 3066, 1732, 1644; mp 290-295°C.

Example 4: Preparation of 2-methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-benzylamide (Ⅴ)

Dissolve 2-methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-3-carboxylic acid (IV) (1g, 0.004mol) in 20mL DMF and cool in ice water to 0°C. In addition, DCC (0.87g, 0.0042mol) and HOBt (0.57g, 0.0042mol) were weighed and dissolved in 50mL CH ₂ Cl ₂ , respectively, added to the reaction solution at 0°C, and stirred. Benzylamine (0.5 mL, 0.004 mol) was added at 0° C. and stirred overnight at room temperature. The reaction solution was washed successively with 5% citric acid, saturated NaHCO ₃ and water, and the organic layer was dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to dryness. Recrystallized from ethanol to obtain 0.72 g of white crystals with a yield of 53.1%.

和7-H)，7.64(1H，d，9-H)，7.80(1H，t，8-H)，8.24(1H，d，6-H)，8.65(1H，s，4-H)。ESI-MS(m/z)：345(M+H⁺)，711(2M+Na⁺)；IR(KBr)cm^-1：3270，1673，1638；m.p.215-216℃  ¹ H-NMR (300MHz, CDCl ₃ ) δ: 2.85 (3H, s, 2-CH ₃ ), 4.70 (2H, d, 2'-CH ₂ -), 6.50 (1H, br, -NH-), 7.34 ～7.46(6H, m, 3'

and 7-H), 7.64 (1H, d, 9-H), 7.80 (1H, t, 8-H), 8.24 (1H, d, 6-H), 8.65 (1H, s, 4-H). ESI-MS (m/z): 345 (M+H ⁺ ), 711 (2M+Na ⁺ ); IR (KBr) cm ^-1 : 3270, 1673, 1638; mp215-216℃

The remaining target compounds are respectively substituted with different groups of o-hydroxyacetophenone and different amines as raw materials, and repeat the steps in Examples 1, 2, 3, and 4 to obtain different 2-methyl-5-oxo-5H -[1]benzopyrano[2,3-b]pyridine-3-amides. All reagents used in the examples are commercially available analytically pure. the

The chemical structures and characterization data of some of the synthesized compounds of the present invention are as follows. the

The structure of the synthesized compound in table 1

2-Methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-methyl-3-amide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-methyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ2.83 (3H, s, 2-CH ₃ ), 3.09 (3H, d, 2'-CH ₃ ), 6.21 (1H, br, -NH-), 7.45 (1H , t, 7-H), 7.60 (1H, d, 9-H), 7.80 (1H, t, 8-H), 8.25 (1H, d, 6-H), 8.62 (1H, s, 4-H ); ESI-MS (m/z): 269 (M+H ⁺ ); IR (KBr) cm ^-1 : 3275, 1673, 1642; mp277-278°C

2-Methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-ethyl-3-amide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-ethyl-3-amide

1H NMR (300MHz, CDCl3) δ: 1.32 (3H, t, 3'-CH ₃ ), 2.84 (3H, s, 2-CH ₃ ), 3.55 (2H, m, 2'-CH ₂ ), 6.05 (1H , br, -NH-), 7.46 (1H, t, 7-H), 7.63 (1H, d, 9-H), 7.82 (1H, t, 8-H), 8.30 (1H, d, 6-H ), 8.65 (1H, s, 4-H). ESI-MS (m/z): 283 (M+H ⁺ ); IR (KBr) cm ^-1 : 3274, 1670, 1637; mp252-254°C

2-Methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-n-propyl-3-amide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-propyl-3-amide

1H NMR (300MHz, CDCl3) δ1.04 (3H, t, 4'-CH ₃ ), 1.68 (2H, m, 3'-CH ₂ ), 2.80 (3H, s, 2-CH ₃ ), 3.48 (2H , m, 2'-CH ₂ ), 6.22 (1H, br, -NH-), 7.48 (1H, t, 7-H), 7.61 (1H, d, 9-H), 7.81 (1H, t, 8 -H), 8.25 (1H, d, 6-H), 8.61 (1H, s, 4-H).ESI-MS (m/z): 295 (M-H+); IR (KBr) cm-1: 3281, 1671, 1634; mp211-212℃

2-Methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-isopropyl-3-amide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-isopropyl-3-amide

¹ H NMR (300MHz, CDCl3) δ: 1.29 (6H, dd, 3'-CH ₃ ×2), 2.84 (3H, s, 2-CH ₃ ), 4.33 (1H, m, 2'-CH), 5.85 (1H, br, -NH-), 7.47 (1H, t, 7-H), 7.64 (1H, d, 9-H), 7.82 (1H, t, 8-H), 8.32 (1H, d, 6 -H), 8.63 (1H, s, 4-H); ESI-MS (m/z): 297 (M+H ⁺ ); IR (KBr) cm-1: 3274, 1671, 1636; mp240-241℃

2-Methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-n-butyl-3-amide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-butyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ1.01 (3H, t, 5'-CH ₃ ), 1.48 (2H, m, 4'-CH ₂ -), 1.66 (2H, m, 3'-CH ₂ - ), 2.83 (3H, s, 2-CH ₃ ), 3.52 (2H, q, 2'-CH ₂ -), 6.10 (1H, br, -NH-), 7.46 (1H, t, 7-H), 7.63(1H, d, 9-H), 7.81(1H, t, 8-H), 8.28(1H, d, 6-H), 8.63(1H, s, 4-H); ESI-MS(m/ z): 311 (M+H ⁺ ); IR (KBr) cm ^-1 : 3285, 1671, 1634; mp193-195°C

2-Methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-benzyl-3-amide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-benzyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ2.85 (3H, s, 2-CH ₃ ), 4.70 (2H, d, 2'-CH ₂ -), 6.50 (1H, br, -NH-), 7.34～ 7.46 (6H, m, Ar-H), 7.64 (1H, d, 9-H), 7.80 (1H, t, 8-H), 8.24 (1H, d, 6-H), 8.65 (1H, s, 4-H); ESI-MS(m/z): 345(M+H) ⁺ , 711(2M+Na) ⁺ ; IR(KBr)cm ^-1 : 3270, 1673, 1638.mp215-216℃

2-Methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

¹ H-NMR (300MHz, CDCl ₃ ) δ2.84 (3H, s, 2-CH ₃ ), 4.65 (2H, d, 2'-CH ₂ -), 6.49 (1H, br, -NH-), 7.34 (4H, m, Ar-H), 7.45 (1H, t, 7-H), 7.61 (1H, d, 9-H), 7.80 (1H, t, 8-H), 8.24 (1H, d, 6 -H), 8.64 (1H, s, 4-H); ESI-MS (m/z): 379 (M+H) ⁺ , 779 (2M+Na) ⁺ ; IR (KBr) cm ^-1 : 3308, 1672, 1637; mp240-241℃

2-Methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-piperidinylamide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-piperidyl amide

¹ H NMR (300MHz, CDCl ₃ ) δ1.55 (2H, m, 4'-CH ₂ -), 1.73 (4H, m, 3', 5'-CH ₂ -×2), 2.71 (3H, s, 2-CH ₃ ), 3.26 (2H, t, 6'-CH ₂ -), 3.81 (2H, t, 2'-CH ₂ -), 7.46 (1H, t, 7-H) 7.62 (1H, d, 9-H), 7.81 (1H, t, 8-H), 8.32 (1H, d, 6-H), 8.50 (1H, s, 4-H); ESI-MS (m/z): 323 (M +H) ⁺ ; IR(KBr)cm ^-1 : 1651, 1635; mp194-195°C

2-Methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-pyrrolylamide

2-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-3-pyrrolidinyl amide

¹ H NMR (300MHz, CDCl ₃ ) δ1.99 (4H, m, 3', 4'-CH ₂ -×2), 2.72 (3H, s, 2-CH ₃ ), 3.28 (2H, t, 2' -CH ₂ -), 3.72 (2H, t, 5'-CH ₂ -), 7.48 (1H, t, 7-H), 7.62 (1H, d, 9-H), 7.80 (1H, t, 8- H), 8.30 (1H, d, 6-H), 8.56 (1H, s, 4-H); ESI-MS (m/z): 309 (M+H ⁺ ), 348 (M+K) ⁺ , 639 (2M+Na ⁺ ); IR (KBr) cm ^-1 : 1664, 1629; mp159-160°C

2-Methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-methyl-3-amide

2-methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-methyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ 2.50 (3H, s, 7-CH ₃ ), 2.84 (3H, s, 2-CH ₃ ), 3.08 (3H, d, 2'-CH ₃ ), 6.11 ( 1H, br, -NH-), 7.52 (1H, d, 9-H), 7.60 (1H, d, 8-H), 8.07 (1H, s, 6-H), 8.65 (1H, s, 4- H); ESI-MS(m/z): 283(M+H) ⁺ ; IR(KBr)cm ^-1 : 3277, 1664, 1643; mp279-281℃.

2-Methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-ethyl-3-amide

2-methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-ethyl-3-amide

¹ H-MR (300MHz, CDCl ₃ ) δ: 1.32 (3H, t, CH ₃ ), 2.50 (3H, s, CH ₃ ), 2.83 (3H, s, CH ₃ ), 3.55 (2H, q, CH ₂ ), 6.04 (1H, br, NH), 7.52 (1H, d, Ar-H), 7.61 (1H, d, Ar-H), 8.08 (1H, s, Ar-H), 8.64 (1H, s, 4-H); ESI-MS(m/z): 297.46(M+H) ⁺ ; IR(KBr)cm ^-1 3274, 1672, 1637; mp258-260℃

2-Methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-propyl-3-amide

2-methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-propyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ: 1.05 (3H, t, 4'-CH ₃ ), 1.71 (2H, m, 3'-CH ₂ ), 2.50 (3H, s, 7-CH ₃ ), 2.82 (3H, s, 2-CH ₃ ), 3.49 (2H, m, 2'-CH ₂ ), 6.11 (1H, br, -NH-), 7.51 (1H, d, 9-H), 7.61 (1H, d, 8-H), 8.07 (1H, s, 6-H), 8.63 (1H, s, 4-H); EI-MS (m/z): 351 (M+K ⁺ ), 311 (M+ H ⁺ ); IR(KBr)cm ^-1 : 3284, 1669, 1635; mp221-222°C

2-Methyl-7-methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-isopropyl-3-amide

2-methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-isopropyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ1.33 (6H, d, 3'-CH ₃ ×2), 2.51 (3H, s, 7-CH ₃ ), 2.83 (3H, s, 2-CH ₃ ), 4.32 (1H, m, 2'-CH), 5.85 (1H, br, -NH-), 7.52 (1H, d, 9-H), 7.61 (1H, d, 8-H), 8.09 (1H, s , 6-H), 8.61 (1H, s, 4-H). ESI-MS (m/z): 311 (M+H ⁺ ). IR (KBr) cm ^-1 : 3298, 1674, 1633.mp254- 256°C.

2-Methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-butyl-3-amide

2-methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-butyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ1.01 (3H, t, 5'-CH ₃ ), 1.48 (2H, m, 4'-CH ₂ -), 1.69 (2H, m, 3'-CH ₂ - ), 2.49 (3H, s, 7-CH ₃ ), 2.77 (3H, s, 2-CH ₃ ), 3.51 (2H, q, 2'-CH ₂ -), 6.32 (1H, br, -NH-) , 7.48(1H, d, 9-H), 7.60(1H, d, 8-H), 8.01(1H, s, 6-H), 8.57(1H, s, 4-H).ESI-MS(m /z): 365(M+2H ⁺ +K ⁺ ).IR(KBr)cm ^-1 : 3298, 1674, 1633.mp220-221℃.

2-Methyl-7-methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-benzyl-3-amide

2-methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-benzyl-3-amide

¹ H NMR (300 MHz, CDCl ₃ ) δ 2.47 (3H, s, 7-CH ₃ ), 2.83 (3H, s, 2-CH ₃ ), 4.69 (2H, d, 2'-CH ₂ ), 6.51 ( 1H, br, -NH-), 7.32～7.41 (5H, m, Ar-H), 7.51 (1H, d, 9-H), 7.61 (1H, d, 8-H), 8.01 (1H, s, 6-H), 8.64 (1H, s, 4-H). ESI-MS (m/z): 739 (2M+Na ⁺ ), 399 (M+K ⁺ ), 359 (M+H ⁺ ).IR (KBr)cm ^-1 : 3270, 1673, 1633.mp248-249℃.

2-Methyl-7-methyl-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

2-methyl-7-methyl-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

¹ H NMR (300 MHz, CDCl ₃ ) δ 2.49 (3H, s, 7-CH ₃ ), 2.84 (3H, s, 2-CH ₃ ), 4.65 (2H, d, 2'-CH ₂ ), 6.53 ( 1H, br, NH), 7.35 (4H, d, Ar-H), 7.51 (1H, d, 9-H), 7.61 (1H, d, 8-H), 8.01 (1H, s, 6-H) , 8.65(1H, s, 4-H).ESI-MS(m/z): 393(M+H ⁺ ).IR(KBr)cm ^-1 : 3277, 1671, 1635.mp245-246℃

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-n-propyl-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-propyl-3-amide

¹ H-NMR (300MHz, DMSO-d ₆ ) δ0.93(t, 3H, CH ₃ ), 1.56(sext, 2H, CH ₂ ), 2.66(s, 3H, CH ₃ ), 3.24(m, 2H, CH ₂ ), 7.77 (d, 1H, J=9.0Hz, Ar-H), 7.94 (dd, 1H, J ₁ =8.7Hz, J ₂ =2.4Hz, Ar-H), 8.07 (s, 1H, Ar -H), 8.46(s, 1H, Ar-H), 8.71(br, 1H, J=5.4Hz, NH).ESI-MS (m/z): 329.63(MH ^- ).IR(KBr)cm ^{- 1} : 3283, 1635, 1671; mp256-258℃

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-isopropyl-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-isopropyl-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ1.33 (6H, d, 2CH ₃ ), 2.85 (3H, s, 2-CH ₃ ), 4.32 (1H, m, CH), 5.80 (1H, br, NH) , 7.60(1H, d, 9-H), 7.76(1H, d, 8-H), 8.28(1H, s, 6-H), 8.62(1H, s, 4-H).ESI-MS(m /z): 331(M+H ⁺ ).IR(KBr)cm ^-1 : 3282, 1672, 1637.mp280-281℃.

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-n-butyl-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-n-butyl-3-amide

¹ H NMR (300 MHz, DMSO-d ₆ ) δ0.92 (t, 3H, CH ₃ ), 1.36 (m, 2H, CH ₂ ), 1.51 (m, 2H, CH ₂ ), 2.67 (s, 3H, CH 2 ) ₃ ), 3.27(m, 2H, CH ₂ ), 7.81(d, 1H, J=9.0Hz, Ar-H), 8.11(d, 1H, J=2.4Hz, Ar-H), 8.48(s, 1H , Ar-H), 8.70 (br, 1H, J=5.4Hz, NH).IR(KBr)cm ^-1 : 3279, 1698, 1673.mp242-244℃.

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-hydroxyethyl-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-hydroxylethyl-3-amide

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 1.24 (m, 2H, CH ₂ ), 2.67 (s, 3H, CH ₃ ), 3.54 (m, 2H, J=5.7 Hz, CH ₂ ), 4.84 ( t, 1H, J = 5.7Hz, OH), 7.81 (d, 1H, J = 9.0Hz, Ar-H), 7.99 (dd, 1H, J ₁ = 9.0Hz, J ₂ = 2.7Hz, Ar-H) , 8.11(d, 1H, J=2.7Hz, Ar-H), 8.56(s, 1H, Ar-H), 8.72(br, 1H, J=5.4Hz, NH).ESI-MS(m/z) : 333.27(M+H ⁺ ).IR(KBr)cm ^-1 : 3274, 1681, 1636.mp249-251℃.

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-benzyl-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-benzyl-3-amide

¹ H NMR (300MHz, DMSO-d ₆ ) δ 2.68 (s, 3H, CH ₃ ), 4.45 (d, 2H, J=5.7Hz, CH ₂ ), 7.28 (dd, 1H, J1=8.4Hz, J ₂ = 4.2Hz, Ar-H), 7.37 (d, 4H, J = 5.2Hz, Ar-H), 7.78 (d, 1H, J = 9.0Hz, Ar-H), 7.94 (dd, 1H, J ₁ =9.0Hz, J ₂ =2.7Hz, Ar-H), 8.07(d, 1H, J=2.7Hz, Ar-H), 8.54(s, 1H, Ar-H), 9.27(t, 1H, J= 5.7Hz, NH).ESI-MS(m/z): 377.35(MH) ^- .IR(KBr)cm ^-1 : 3269, 1675, 1634.mp268-270℃.

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-fluorobenzyl)-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-flurobenzyl)-3-amide

¹ H NMR (300MHz, DMSO-d ₆ ) δ2.68 (s, 3H, CH ₃ ), 4.48 (d, 2H, J ₂ =6.0Hz, CH ₂ ), 7.16-7.22 (m, 2H, Ar-H ), 7.39-7.44 (m, 2H, Ar-H), 7.80 (d, 1H, J=9.0Hz, Ar-H), 7.96 (dd, 1H, J ₁ =9.0Hz, J ₂ =2.7Hz, Ar -H), 8.10(d, 1H, J=2.4Hz, Ar-H), 8.56(s, 1H, Ar-H), 9.25(t, 1H, J=6.0Hz, NH).ESI-MS(m /z): 395.62(MH) ^- .IR(KBr)cm ^-1 : 3270, 1677, 1635.mp269℃(dec).

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ2.86(s, 3H, CH ₃ ), 4.64(d, 2H, J=5.7Hz, CH ₂ ), 6.28(br, 1H, NH), 7.32-7.39(m , 4H, Ar-H), 7.59(d, 1H, J=8.7Hz, Ar-H), 7.75(dd, 1H, J ₁ =9.0Hz, J ₂ =2.4Hz, Ar-H), 8.25(d , 1H, J=2.7Hz, Ar-H), 8.65 (s, 1H, Ar-H).ESI-MS(m/z): 411.64(MH) ^- .IR(KBr)cm ^-1 : 3273, 1674 , 1634.mp248℃(dec).

2-Methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-methylbenzyl)-3-amide

2-methyl-7-chloro-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-methylbenzyl)-3-amide

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 2.29 (s, 3H, CH ₃ ), 2.68 (s, 3H, CH ₃ ), 4.44 (d, 2H, J=5.7 Hz, CH ₂ ), 7.17 ( d, 2H, J=8.1Hz, Ar-H), 7.26(d, 2H, J=8.1Hz, Ar-H), 7.81(d, 1H, J=9.0Hz, Ar-H), 7.97(dd, 1H, J ₁ =9.0Hz, J ₂ =2.7Hz, Ar-H), 8.11(d, 1H, J=2.7Hz, Ar-H), 8.54(s, 1H, Ar-H), 9.23(br, 1H, NH).ESI-MS(m/z): 391.74(MH) ^- .IR(KBr)cm ^-1 : 3277, 1675, 1635.mp261℃(dec).

2-Methyl-7-methoxy-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-(4-n-butyl)-3-amide

2-methyl-7-methoxy-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-n-butyl)-3-amide

¹ H NMR (300MHz, CDCl ₃ ) δ0.92 (t, 3H, CH ₃ ), 1.40 (m, 2H, CH ₂ ), 1.52 (m, 2H, CH ₂ ), 2.67 (s, 3H, CH ₃ ) , 3.26(m, 2H, CH ₂ ), 3.95(s, 3H, OCH ₃ ), 7.10(dd, 1H, J ₁ =8.7Hz, J ₂ =2.4Hz, Ar-H), 7.26(d, 1H, J=2.4Hz, Ar-H), 8.09(d, 1H, J=9.0Hz, Ar-H), 8.45(s, 1H, Ar-H), 8.68(br, 1H, J=5.4Hz, NH) .ESI-MS(m/z): 341.66(M+H) ⁺ .IR(KBr)cm ^-1 : 3280, 1694, 1634.mp248-250℃.

2-Methyl-7-methoxy-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-benzyl-3-amide

2-methyl-7-methoxy-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-benzyl)-3-amide

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 2.68 (s, 3H, CH ₃ ), 3.94 (s, 3H, OCH ₃ ), 4.50 (d, 2H, J=5.7 Hz, CH ₂ ), 7.09 ( dd, 1H, _J1 = 9.0Hz, J2 = 2.4Hz, Ar-H), 7.24 (d, 1H, J = 2.4Hz _, Ar-H), 7.28 (d, 1H, J = 4.5Hz, Ar- H), 7.37(d, 4H, J=4.5Hz, Ar-H), 8.08(d, 1H, J=9.0Hz, Ar-H), 8.53(s, 1H, Ar-H), 9.23(br, 1H, NH).ESI-MS(m/z): 373.72(MH) ^- .IR(KBr)cm ^-1 : 3270, 1663, 1634.mp268-270℃.

2-Methyl-7-methoxy-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-fluorobenzyl)-3-amide

2-methyl-7-methoxy-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-flurobenzyl)-3-amide

¹ H NMR (300MHz, DMSO-d ₆ ) δ2.67 (s, 3H, CH ₃ ), 3.95 (s, 3H, OCH ₃ ), 4.47 (d, 2H, J=6.0Hz, CH ₂ ), 7.08- 7.26(m, 4H, Ar-H), 7.39-7.43(m, 2H, Ar-H), 8.09(d, 1H, J=9.0Hz, Ar-H), 8.53(s, 1H, Ar-H) , 9.23 (br, 1H, J=6.0Hz, NH).EI-MS(m/z): 393.72(M+H) ⁺ .IR(KBr)cm ^-1 : 3292, 1716, 1662.mp295-297℃ .

2-Methyl-7-methoxy-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

2-methyl-7-methoxy-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-chlorobenzyl)-3-amide

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 2.67 (s, 3H, CH ₃ ), 3.95 (s, 3H, OCH ₃ ), 4.48 (d, 2H, J=6.0 Hz, CH ₂ ), 7.10 ( dd, 1H, _J1 = 9.0Hz, J2 = 2.4Hz, Ar-H) _, 7.25 (d, 1H, J = 2.4Hz, Ar-H), 7.38-7.45 (m, 4H, Ar-H), 8.09 (d, 1H, J = 8.7Hz, Ar-H), 8.54 (s, 1H, Ar-H), 9.24 (br, 1H, J = 6.0Hz, NH). ESI-MS (m/z): 407.56MH) ^- .IR(KBr)cm ^-1 : 3294, 1716, 1662.mp270℃(dec).

2-Methyl-7-methoxy-5-oxo-5H-[1]benzopyran[2,3-b]pyridine-N-(4-methylbenzyl)-3-amide

2-methyl-7-methoxy-5-oxo-5H-[1]benzopyrano[2,3-b]pyridine-N-(4-methylbenzyl)-3-amide

¹ H NMR (300MHz, DMSO-d ₆ ) δ 2.29 (s, 3H, CH ₃ ), 2.67 (s, 3H, CH ₃ ), 3.95 (s, 3H, OCH ₃ ), 4.44 (d, 2H, J =5.7Hz, _CH2 ), 7.10(dd, 1H, _J1 =9.0Hz, _J2 =2.4Hz, Ar-H), 7.17(d, 2H, J=7.8Hz, Ar-H), 7.25(d , 2H, J=3.9Hz, Ar-H), 7.26(d, 1H, J=3.9Hz, Ar-H), 8.09(d, 1H, J=9.0Hz, Ar-H), 8.50(s, 1H , Ar-H), 9.19 (br, 1H, NH). EI-MS (m/z): 389.61 (M+H) ⁺ .IR (KBr) cm ^-1 : 3257, 1662, 1637.mp275-277℃ .

Embodiment 5: the cytotoxic effect of the compound of the present invention

(1) Experimental materials and methods:

1. Experimental materials

(1) Experimental strains

In this experiment, three human cancer cell lines were used as screening objects, and the cancer cell lines were all provided by the Pharmacology Laboratory of Shanghai Institute of Pharmaceutical Industry. the

1) Human lung cancer cell A549

2) Human colon cancer cell Colo205

3) Human breast cancer cell MDA-MB-435

(2) The culture medium is RPMI1640+15%NBS+double antibody. The model of the automatic microplate reader used is: WellscanMK-2, the manufacturer: Labsystems. Imported 96-well culture plates, etc. the

2. Experimental method

In vitro anti-tumor activity was determined by MTT method. the

Sample preparation: After dissolving with DMSO (Merck), add PBS (-) to make a 1000 μg/ml solution or a homogeneous suspension, and then dilute with DMSO-containing PBS (-). the

Culture medium: RPMI640+15%NBS+double antibody

Positive control drug: hydroxycamptothecin

The experimental steps are as follows:

Add 1000 μl of cell suspension with a concentration of 4-5×10 ⁴ cells/ml to each well of a 96-well culture plate, and place in a 37° C., 5% CO ₂ incubator. After 24 hours, add the sample solution, 10 μl/well, set up duplicate wells, and act at 37°C and 5% CO ₂ for 72 hours. Add 20 μl of 5 mg/ml MTT solution to each well, and add the dissolving solution after 4 hours of action, 100 μl/well, put it in the incubator, measure the 570nmOD value with MK-2 automatic microplate reader after dissolving.

(2) Experimental results

Table 2.29 Compounds Inhibitory Effects on Human Tumor Cell Proliferation in Vitro

Example 6: Topoisomerase I inhibitory activity test

Using literature methods (Kingsbury WD, Boehm JC, Jakas DR, et al. J Med Chem, 1991, 34, 98-107), some compounds were tested for topoisomerase I inhibitory activity, and they were found to be in the range of 0.01-100uM It has good activity of inhibiting Top1. the

In summary, the compounds of the present invention have certain anti-tumor effects. It is particularly worth pointing out that these compounds show good selective toxicity to metastatic breast cancer cells. Tumor drugs have laid the foundation and have good development value. The 1-azoxanthone-3-carboxamide compound of the present invention represents a class of compounds with a new structure and antitumor activity, which opens up a new approach and direction for in-depth research and development of new antitumor drugs. the

Claims (6)

1. a class 1-azepine xanthone-3-Carbox amide is selected from:

2. the preparation method of 1-azepine xanthone claimed in claim 1-3-Carbox amide may further comprise the steps:

(1) preparation 3-cyano group-4-benzopyrone (II)

First DMF and phosphorus oxychloride are reacted half an hour at 0 ℃, add again the hydroxy acetophenone (I) that replaces, add again oxammonium hydrochloride behind the stirring at room reaction 4h and continue to react 6h, generate 3-cyano group-4-benzopyrone (II);

(2) preparation 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxylic acid ethyl ester (III)

3-cyano group-4-benzopyrone (II), methyl aceto acetate and piperidines are refluxed in ethanol, generate 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxylic acid ethyl ester (III);

(3) preparation 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxylic acid (IV)

With 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxylic acid ethyl ester (III) is at 50%H ₂SO ₄Reflux in-AcOH the mixing acid, generate 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxylic acid (IV);

(4) preparation 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxamide (V)

With 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxylic acid (IV) is dissolved among the DMF, the dichloromethane solution that under 0 ℃, adds respectively DCC and HOBt, add amine after the stirred for several minute, stirring at room reaction 24h, generate 2-methyl-5-oxo-5H-[1] chromene [2,3-b] pyridine-3-carboxamide (V);

Wherein, definition in each symbol in the formula such as the claim 1.

3. the 1-azepine xanthone of a class shown in general formula (I)-3-Carbox amide or its pharmacy acceptable salt application in the preparation antitumor drug:

Wherein, R ₁The position of substitution can be positioned at 6 to 9, can be single, double or polysubstituted, R ₁Be the arbitrary class in the following groups: a) hydrogen; B) the straight or branched alkoxyl group of C1-8; C) the straight or branched alkyl of C1-8; D) halogen; M) methylene-dioxy;

R ₂, R ₃Represent independently following groups: the alkyl of the straight or branched of hydrogen, C1-8, replacement or unsubstituted benzyl, alkoxyl group, nitro, amino, the hydroxyl of the straight or branched of the described alkyl that is substituted by the straight or branched of halogen, C1-8, C1-8; Or R ₂, R ₃Form together by 3 to 7 CH ₂The chain that the unit connects to form;

R ₄Straight or branched alkyl for hydrogen, C1-8.

4. according to the application of formula claimed in claim 3 (I) compound or its salt at the preparation antitumor drug, it is characterized in that tumour is mammary cancer.

5. according to the application of formula claimed in claim 3 (I) compound or its salt at the preparation antitumor drug, it is characterized in that tumour is metastatic breast cancer.

6. a composition contains any 1-azepine xanthone-3-Carbox amide claimed in claim 1 or its pharmacy acceptable salt and reaches the carrier of pharmaceutically accepting.