Application of N2, N3-di-tert-butylquinoxaline-2, 3-diamine derivative in preparation of antitumor drug composition

Abstract

The invention belongs to the technical field of antitumor drugs, and particularly discloses an application of a N2, N3-di-tert-butylquinoxaline-2, 3-diamine derivative in preparation of an antitumor drug composition. The N2, N3-di-tert-butylquinoxaline-2, 3-diamine derivative provided by the invention has a remarkable inhibition effect on human colon cancer cells HCT116, human liver cancer cells HepG-2, human lung cancer cells A549, human prostate cancer cells PC3, human cervical cancer cells Hela, human glial cancer cells U87 and human malignant breast cancer cells 4T1, shows excellent anti-tumor characteristics, and can be used for preparing medicines for treating tumors. The compound can be used as an anti-tumor lead drug molecule, and has a good development and application prospect in the aspect of developing anti-tumor drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of anti-tumor drugs, and relates to the application of a di-tert-butylquinoxaline diamine derivative in the preparation of an anti-tumor drug composition, and specifically relates to N 2 ,N 3 -Application of di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor drug compositions. Background Art

[0002] Quinoxaline derivatives do have broad application prospects in medicinal chemistry, especially in the development of anti-tumor drugs. These compounds have attracted the attention of many medicinal chemists due to their unique structure and multiple biological activities, such as antibacterial, anticancer, analgesic, antimalarial, antitumor, anti-epileptic, anticonvulsant, anti-tuberculosis, antiproliferative and anti-inflammatory.

[0003] As a member of the benzodiazine family, quinoxaline has a skeleton structure that gives it a variety of biological activities. In terms of anti-tumor effects, quinoxaline derivatives can inhibit the growth and spread of tumor cells through a variety of mechanisms, including but not limited to: (1) Cell cycle arrest: Some quinoxaline derivatives can interfere with the cell cycle of tumor cells, causing them to arrest at a specific stage, thereby inhibiting cell proliferation. (2) Apoptosis: By inducing apoptosis of tumor cells, quinoxaline derivatives can effectively eliminate cancer cells in the body. (3) Inhibition of angiogenesis: Angiogenesis is an important process in tumor growth and metastasis. Quinoxaline derivatives can inhibit tumor growth by inhibiting angiogenesis. (4) DNA embedding and damage: Some quinoxaline derivatives can embed into DNA, interfering with the normal replication and transcription of DNA, thereby damaging DNA and causing tumor cell death.

[0004] At present, quinoxaline derivatives have been widely studied in anti-tumor drugs, for example: (1) Echinomycin and its analogs: Echinomycin is a natural antibiotic with two terminal quinoxaline-2-carboxylic acid groups that can bind to the cytosine-guanine (CG) base pairs of DNA and act as a double intercalator. By further modifying Echinomycin, researchers have developed a variety of new quinoxaline antibiotics that show excellent anti-tumor activity. (2) Angiogenesis inhibitors: Some quinoxaline derivatives have been developed as angiogenesis inhibitors, such as DCQ and BPQ, which show significant activity against a variety of tumor cells such as Lewis lung cancer and EMT-6 breast cancer cells. (3) Kinase inhibitors: Protein kinases play a key role in a variety of cellular functions such as cell cycle, proliferation, and DNA damage repair. Quinoxaline derivatives have been developed as inhibitors of a variety of kinases, such as inhibitors of c-Src, c-Abl, PI3K / AKT and other kinases. These inhibitors show good application prospects in cancer treatment.

[0005] With the continuous deepening of the research on quinoxaline derivatives, their application prospects in the development of anti-tumor drugs will be broader. Quinoxaline derivatives do have great potential in the treatment of various tumors and are worthy of further in-depth research and development.

[0006] There is currently no N 2 ,N 3 -Related research reports on the anti-tumor biological activity or clinical application of di-tert-butylquinoxaline-2,3-diamine derivatives. Summary of the invention

[0007] The present invention provides a novel application of di-tert-butylquinoxaline diamine derivatives in the preparation of anti-tumor drug compositions.

[0008] The first object of the present invention is to provide N 2 ,N 3 -The chemical structure of di-tert-butylquinoxaline-2,3-diamine derivative is as follows:

[0009]

[0010] Among them, R1 is one of a hydrogen group and a methyl group; R2 is one of a hydrogen group, a fluoro group, a chloro group and a methyl group; R3 is one of a hydrogen group, a methoxy group, a pentyl group, a methyl group, a trifluoromethoxy group, a fluoro group, a chloro group and a bromo group; R4 is one of a hydrogen group and a methyl group.

[0011] In the present invention, it is further described that the tumor includes but is not limited to colon cancer, liver cancer, lung cancer, prostate cancer, cervical cancer, glial cancer and breast cancer.

[0012] In the present invention, it is further described that R1 is a hydrogen group (H) and a methyl group One of: R2 is hydrogen (H), fluorine (F), chlorine (Cl) and methyl One of: R3 is hydrogen (H), methoxy Pentyl methyl trifluoromethoxy One of fluoro (F), chloro (Cl) and bromo (Br); R4 is hydrogen (H) and methyl One of them.

[0013] In the present invention, it is further described that the above N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives are catalytically synthesized by the following steps, specifically:

[0014]

[0015] Take a 25ml reaction tube, add o-phenylenediamine derivative 1 (0.1mmol), tert-butyl isonitrile 2 (0.25mmol), Pd(PhCN)2Cl2 (10mol%), carbon tetrabromide (1.0equiv), silver nitrate (2.0equiv), pyridine (3.0equiv) and dimethyl sulfoxide (1mL) into the reaction tube in sequence, and continue the reaction at 100℃ for 3 hours. After the reaction is completed, add 10mL of water to quench the reaction, extract with ethyl acetate three times (3×10mL), recover the organic layer, and concentrate it with a rotary evaporator. The crude product is separated by column chromatography to obtain the target product 3, and the developing solvent used is n-hexane / ethyl acetate (5 / 1). The product is then confirmed by nuclear magnetic resonance and high-resolution mass spectrometry characterization methods.

[0016] The second object of the present invention is to provide N 2 ,N 3 -Application of di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor drug compositions.

[0017] In the present invention, it is further described that the N 2 ,N 3 -di-tert-butylquinoxaline-2,3-diamine derivatives are used in the preparation of anti-tumor drug compositions, wherein N 2 ,N 3 -di-tert-butylquinoxaline-2,3-diamine derivatives as candidate drugs or lead compounds for treating tumors, using N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives have inhibitory activity against a variety of tumor cells, laying the foundation for the development of new anti-tumor drugs.

[0018] In the present invention, it is further described that the N 2 ,N 3 - Use of di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor pharmaceutical compositions, wherein the therapeutic object of the pharmaceutical composition is a mammal or a human.

[0019] In the present invention, it is further described that the N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor pharmaceutical compositions, wherein the pharmaceutical compositions are N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives are used as the main component, and pharmaceutically acceptable excipients or auxiliary components are added to prepare clinically acceptable pharmaceutical preparations.

[0020] In the present invention, it is further described that the N 2 ,N 3 The content of the di-tert-butylquinoxaline-2,3-diamine derivative in the pharmaceutical composition is usually 0.1-95.0% (w / w).

[0021] In the present invention, it is further described that the N 2 ,N 3 -Application of di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor pharmaceutical compositions, wherein the pharmaceutical preparations include oral preparations and injection preparations.

[0022] In the present invention, it is further described that the N 2 ,N 3 - Use of di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor pharmaceutical compositions, wherein the oral preparation is an oral capsule and the injection preparation is an intravenous injection.

[0023] Generally speaking, as a drug, it is clinically used after being prepared into a preparation. The pharmaceutical composition of the present invention can be prepared according to methods known in the art as a pharmaceutical composition. The pharmaceutical composition of the present invention can be combined with one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants to prepare any dosage form suitable for human or animal use.

[0024] The pharmaceutical composition of the present invention or a pharmaceutical composition containing the same can be administered in a unit dosage form, and the administration route can be enteral or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0025] The dosage form can be a liquid dosage form, a solid dosage form or a semisolid dosage form. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o / w types, w / o types and multiple emulsions), suspensions, injections (including water injections, powder injections and infusions), eye drops, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, micropills, dropping pills, suppositories, films, patches, aerosols (powders), sprays, etc.; semisolid dosage forms can be ointments, gels, pastes, etc.

[0026] The pharmaceutical composition of the present invention can be made into a common preparation, or a sustained-release preparation, a controlled-release preparation, a targeted preparation, and various microparticle delivery systems. In order to make the pharmaceutical composition of the present invention into tablets, various excipients known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, and glidants. The diluent may be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agent may be water, ethanol, isopropanol, etc.; the adhesive may be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinyl pyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0027] The tablets can be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

[0028] In order to prepare the dosing unit into a capsule, the active ingredient pharmaceutical composition of the present invention can be mixed with a diluent and a glidant, and the mixture can be directly placed in a hard capsule or a soft capsule. The active ingredient pharmaceutical composition of the present invention can also be first prepared into granules or pellets with a diluent, a binder, and a disintegrant, and then placed in a hard capsule or a soft capsule. The diluents, binders, wetting agents, disintegrants, and glidants used to prepare the tablets of the pharmaceutical composition of the present invention can also be used to prepare the capsules of the pharmaceutical composition of the present invention.

[0029] To prepare the pharmaceutical composition of the present invention into an injection, water, ethanol, isopropanol, propylene glycol or a mixture thereof can be used as a solvent and an appropriate amount of a solubilizer, a cosolvent, a pH adjuster, and an osmotic pressure regulator commonly used in the art can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc.; the pH adjuster can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. If a lyophilized powder injection is prepared, mannitol, glucose, etc. can also be added as a support agent.

[0030] Furthermore, if necessary, colorants, preservatives, perfumes, flavoring agents or other additives may be added to the pharmaceutical preparations.

[0031] Compared with the prior art, the present invention has the following advantages:

[0032] 1. The application of the di-tert-butylquinoxaline diamine derivatives developed by the present invention in the preparation of anti-tumor drug compositions has significant medical value and broad application prospects. In particular, N 2 ,N 3 -The application exploration of di-tert-butylquinoxaline-2,3-diamine derivatives in the field of anti-tumor drugs has indeed shown great potential and value. This type of derivative has opened up a new direction for the research and development of anti-tumor drugs due to its unique chemical structure and biological activity. The R1, R2, R3 and R4 groups introduced into the derivative can optimize its selectivity, affinity and metabolic stability by precisely regulating these groups, thereby enhancing its anti-tumor effect and reducing side effects. 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives are synthesized from o-phenylenediamine derivatives and tert-butyl isonitrile by catalysis of Pd(PhCN)2Cl2 (10mol%), carbon tetrabromide (1.0equiv), silver nitrate (2.0equiv) and pyridine (3.0equiv). Not only are the reaction conditions mild, but the required raw materials and catalysts are relatively cheap and easy to obtain, which is of great significance for future large-scale industrial production. In addition, this method may also have high atom economy and environmental friendliness, which is in line with the development trend of green chemistry. Based on the current research results, further research on N 2 ,N 3 -The preclinical and clinical trial research and development of di-tert-butylquinoxaline-2,3-diamine derivatives are expected to provide cancer patients with safer and more effective treatment options, improve the quality of life of cancer patients, and reduce the social medical burden. 2 ,N 3 -The application research of di-tert-butylquinoxaline-2,3-diamine derivatives in the field of anti-tumor drugs not only brings new hope to cancer patients, but also provides new ideas and methods for drug research and development.

[0033] 2. The present invention verifies N through experiments 2 ,N 3 -Application of di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor drug compositions, with a wide range of anti-tumor effects, 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives showed good inhibitory activity against human colon cancer cells HCT116, human liver cancer cells HepG-2, human lung cancer cells A549, human prostate cancer cells PC3, human cervical cancer cells Hela, human glial cancer cells U87, and human malignant breast cancer cells 4T1. The results showed that N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives have better inhibitory effects on tumor cells than the positive drug 5-fluorouracil. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 is the N of the present invention 2 ,N 3 -Chemical structure diagram of di-tert-butylquinoxaline-2,3-diamine derivatives. DETAILED DESCRIPTION

[0035] The specific implementation modes of the present invention are further described below in conjunction with examples.

[0036] Embodiment 1:

[0037] N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives are obtained by the following synthetic route, and the specific steps are as follows:

[0038]

[0039] Take a 25ml reaction tube, add o-phenylenediamine derivative 1 (0.1mmol), tert-butyl isonitrile 2 (0.25mmol), Pd(PhCN)2Cl2 (10mol%), carbon tetrabromide (1.0equiv), silver nitrate (2.0equiv), pyridine (3.0equiv) and dimethyl sulfoxide (1mL) into the reaction tube in sequence, and continue the reaction at 100℃ for 3 hours. After the reaction is completed, add 10mL of water to quench the reaction, extract with ethyl acetate three times (3×10mL), recover the organic layer, and concentrate it with a rotary evaporator. The crude product is separated by column chromatography to obtain the target product 3, and the developing solvent used is n-hexane / ethyl acetate (5 / 1). The product is then confirmed by nuclear magnetic resonance and high-resolution mass spectrometry characterization methods.

[0040] Embodiment 2:

[0041] Compound 1 was prepared by the following synthesis route, and the specific steps are as follows: take a 25 ml reaction tube, add o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 1, whose structural formula is synthesized by the following technical route), with a yield of 92%.

[0042] The technical route for preparing compound 1 is as follows:

[0043]

[0044] The characterization data of compound 1 are as follows: 1 H NMR (500MHz, CDCl3, δppm)7.66-7.63(m,2H),7.32-7.31(m,2H),4.09(s,2H),1.60(s,18H); 13 C NMR(126MHz, CDCl3, δppm)144.0,136.4,125.9,124.3,52.2,29.0.HRMS(ESI)m / z:calcd for C 16 H 25 N4[M+H] + 273.2079; found 273.2076.

[0045] Embodiment 3:

[0046] Compound 2 was prepared by the following synthesis route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-methoxy-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 2, whose structural formula is synthesized by the following technical route), with a yield of 80%.

[0047] The technical route for preparing compound 2 is as follows:

[0048]

[0049] The characterization data of compound 2 are as follows: 1H NMR(500MHz,DMSO,δppm)7.27(d,J=8.7Hz,1H),6.84(d,J=2.8Hz,1H),6.78-6 .76(m,1H),6.26(s,1H),6.11(s,1H),3.77(s,3H),1.52(s,9H),1.50(s,9H); 13 C NMR(126MHz,DMSO,δppm)156.4,143.9,142.2,136.7,130.6,126.0,113.4,106.3,55.6,52.3,52.1,29.3,29.2.HRMS(ESI)m / z:calcd for C 17 H 27 N4O[M+H] + 303.2185; found 303.2191.

[0050] Embodiment 4:

[0051] Compound 3 was prepared by the following synthesis route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-tert-butyl o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 3, whose structural formula is synthesized by the following technical route), with a yield of 61%.

[0052] The technical route for preparing compound 3 is as follows:

[0053]

[0054] The characterization data of compound 3 are as follows: 1 H NMR (400MHz, DMSO, δppm) 7.30 (d, J = 8.1Hz, 2H), 7.19 (d, J = 8.5Hz, 1H), 6.24 (s, 1H), 6.22 (s, 1H), 1.52 (s, 9H), 1.51 (s, 9H), 1.30 (s, 9H); 13 CNMR(101MHz,DMSO,δppm)146.0,143.6,143.3,135.5,133.8,124.8,121.5,121.0,52.2,52.1,34.6,31.9,29.3,29.2.HRMS(ESI)m / z:calcd for C 20 H33 N4[M+H] + 329.2710; found 329.2705.

[0055] Embodiment 5:

[0056] Compound 4 was prepared by the following synthesis route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-methyl-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 4, whose structural formula is synthesized by the following technical route), with a yield of 86%.

[0057] The technical route for preparing compound 4 is as follows:

[0058]

[0059] The characterization data of compound 4 are as follows: 1 H NMR (400MHz, CDCl3, δppm) 7.49 (d, J = 8.2 Hz, 1H), 7.41 (s, 1H), 7.12 (d, J = 8.1 Hz, 1H), 4.05 (s, 1H), 3.97 (s, 1H), 2.45 (s, 3H), 1.56 (s, 9H), 1.56 (s, 9H). 13 C NMR (101MHz, CDCl3, δppm)144.1,143.5,138.5,136.2,134.1,125.9,125.6,125.5,52.1,29.0,28.9,21.3.HRMS(ESI)m / z:calcd for C 17 H 27 N4[M+H] + 287.2236; found 288.2237.

[0060] Embodiment 6:

[0061] Compound 5 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 3-methyl-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 5, whose structural formula is synthesized by the following technical route), with a yield of 47%.

[0062] The technical route for preparing compound 5 is as follows:

[0063]

[0064] The characterization data of compound 5 are as follows: 1 H NMR (400MHz, CDCl3, δppm) 7.49-7.46 (m, 1H), 7.21 (d, J = 7.2Hz, 1H), 7.17 (d, J = 6.8Hz, 1H), 4.06 (s, 2H), 2.64 (s, 3H), 1.60 (s, 9H), 1.57 (s, 9H). 13 C NMR (101MHz, CDCl3, δppm)143.6,142.8,136.1,135.1,133.9,124.9,123.9,52.1,52.0,29.0,28.7,17.6.HRMS(ESI)m / z:calcd for C 17 H 27 N4[M+H] + 287.2236; found 287.2241.

[0065] Embodiment 7:

[0066] Compound 6 was prepared by the following synthesis route, and the specific steps are as follows: take a 25 ml reaction tube, add 4,5-dimethyl-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 6, whose structural formula is synthesized by the following technical route), with a yield of 87%.

[0067] The technical route for preparing compound 6 is as follows:

[0068]

[0069] The characterization data of compound 6 are as follows: 1 H NMR (400MHz, CDCl3, δppm) 7.39 (s, 2H), 4.00 (s, 2H), 2.36 (s, 8H), 1.56 (s, 18H). 13 C NMR(101MHz, CDCl3, δppm)143.7,134.7,133.4,125.9,52.0,29.0,19.7.HRMS(ESI)m / z:calcd for C 18 H 29 N4[M+H] + 301.2392; found 301.2391.

[0070] Embodiment 8:

[0071] Compound 7 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-trifluoromethoxy-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 7, whose structural formula is synthesized by the following technical route), with a yield of 30%.

[0072] The technical route for preparing compound 7 is as follows:

[0073]

[0074] The characterization data of compound 7 are as follows: 1 H NMR (400MHz, CDCl3, δppm) 7.57 (d, J = 8.8 Hz, 1H), 7.44 (d, J = 1.4 Hz, 1H), 7.15-7.11 (m, 1H), 4.18 (s, 1H), 4.08 (s, 1H), 1.56 (s, 9H), 1.56 (s, 9H). 13 C NMR (101MHz, CDCl3, δppm) 145.7, 144.4, 143.9, 136.5, 126.8, 122.0, 119.4, 117.6, 117.3, 52.4, 52.3, 28.9, 28.8. 19 F NMR(376MHz, CDCl3, δppm)-58.20.HRMS(ESI)m / z:calcdfor C 17 H24 N4OF3[M+H] + 357.1897; found 357.1902.

[0075] Embodiment 9:

[0076] Compound 8 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-fluoro-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 8, whose structural formula is synthesized by the following technical route), with a yield of 59%.

[0077] The technical route for preparing compound 8 is as follows:

[0078]

[0079] The characterization data of compound 8 are as follows: 1 H NMR (400MHz, CDCl3, δppm)7.55-7.51(m,1H),7.26-7.23(m,1H),7.04-7.00(m,1H),4.19(s,1H),3.95(s,1H),1.55(s,18H). 13 C NMR (101MHz, CDCl3, δppm) 160.1 (d, J = 241.39Hz), 144.5, 143.3, 137.0 (d, J = 10.1Hz), 132. 9,126.4(d,J=60.6Hz),112.6(d,J=10.1Hz),110.3(d,J=10.1Hz),52.3,52.2,29.0,28.9. 19 FNMR(376MHz,CDCl3,δppm)-123.64.HRMS(ESI)m / z:calcd for C 16 H 24 N4F[M+H] + 291.1985; found 291.1987.

[0080] Embodiment 10:

[0081] Compound 9 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-chloro-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 9, whose structural formula is synthesized by the following technical route), with a yield of 79%.

[0082] The technical route for preparing compound 9 is as follows:

[0083]

[0084] The characterization data of compound 9 are as follows: 1 H NMR (500MHz, CDCl3, δppm) 7.62 (d, J = 2.2 Hz, 1H), 7.53 (d, J = 8.6 Hz, 1H), 7.26-7.23 (m, 1H), 4.20 (s, 2H), 1.59 (s, 18H); 13 C NMR (126MHz, CDCl3, δppm)144.3,143.8,136.9,134.8,129.2,126.9,125.0,124.6,52.4,52.3,28.9,28.8.HRMS(ESI)m / z:calcd for C 16 H 24 N4Cl[M+H] + 307.1689; found 307.1695.

[0085] Embodiment 11:

[0086] Compound 10 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-bromo-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 10, whose structural formula is synthesized by the following technical route), with a yield of 79%.

[0087] The technical route for preparing compound 10 is as follows:

[0088]

[0089] The characterization data of compound 10 are as follows: 1 H NMR (500MHz, CDCl3, δppm) 7.75 (d, J = 2.2 Hz, 1H), 7.44 (d, J = 8.6 Hz, 1H), 7.35-7.32 (m, 1H), 4.13 (s, 1H), 4.07 (s, 1H), 1.55 (s, 9H), 1.55 (s, 9H); 13 C NMR (126MHz, CDCl3, δppm)144.2,143.9,137.3,135.1,128.1,127.2,117.3,117.0,52.4,28.9,28.8.HRMS(ESI)m / z:calcd for C 16 H 24 N4Br[M+H] + 351.1184; found353.1168,353.1186.

[0090] Embodiment 12:

[0091] Compound 11 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4,5-difluoro-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 11, whose structural formula is synthesized by the following technical route), with a yield of 40%.

[0092] The technical route for preparing compound 11 is as follows:

[0093]

[0094] The characterization data of compound 11 are as follows: 1 H NMR (400MHz, DMSO, δppm) 7.27 (t, J = 10.3Hz, 2H), 6.50 (s, 2H), 1.49 (s, 18H). 13 C NMR (101MHz, DMSO, δppm) 147.0 (dd, J = 226.4, 16.8Hz), 143.6, 132.4 (t, J = 6.0Hz), 111.38 (dd, J = 11.5, 6.8Hz), 52.5, 29.1. 19FNMR(376MHz,DMSO,δppm)-154.18.HRMS(ESI)m / z:calcd for C 16 H 23 N4F2Na[M+H] + 309.1895; found 309.1891.

[0095] Embodiment 13:

[0096] Compound 12 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4,5-dichloro-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100 ° C for 3 hours to obtain the target compound (compound 12, whose structural formula is synthesized by the following technical route), with a yield of 35%.

[0097] The technical route for preparing compound 12 is as follows:

[0098]

[0099] The characterization data of compound 12 are as follows: 1 H NMR (400MHz, DMSO, δppm) 7.50 (s, 2H), 6.69 (s, 2H), 1.50 (s, 18H); 13 C NMR(101MHz,DMSO,δppm)144.1,135.6,125.6,125.052.729.0.HRMS(ESI)m / z:calcd for C 20 H 18 N3ClNa[M+H] + 341.1303; found 341.1300.

[0100] Embodiment 14:

[0101] Compound 13 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-chloro-5-methyl-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 13, whose structural formula is synthesized by the following technical route), with a yield of 64%.

[0102] The technical route for preparing compound 13 is as follows:

[0103]

[0104] The characterization data of compound 13 are as follows: 1 H NMR (400MHz, DMSO, δppm)7.34(s,1H),7.31(s,1H),6.43(s,2H),2.35(s,3H),1.50(s,9H),1.50(s,9H). 13 C NMR(101MHz,DMSO,δppm)143.7,143.6,135.1,134.9,130.1,128.1,126.7,124.1,52.4,29.1,20.0.HRMS(ESI)m / z:calcdfor C 17 H 26 N4Cl[M+H] + 321.1849; found 321.1846.

[0105] Embodiment 15:

[0106] Compound 14 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 4-bromo-5-methyl-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 14, whose structural formula is synthesized by the following technical route), with a yield of 45%.

[0107] The technical route for preparing compound 14 is as follows:

[0108]

[0109] The characterization data of compound 14 are as follows: 1 H NMR (400MHz, DMSO, δppm)7.52(s,1H),7.32(s,1H),6.44(s,1H),6.43(s,1H),2.37(s,3H),1.51(s,9H),1.50(s,9H). 13 C NMR(101MHz,DMSO,δppm)143.8,143.6,135.4,135.3,131.6,127.4,126.6,118.4,52.4,52.4,29.1,29.1,22.6.HRMS(ESI)m / z:calcd for C 17 H 26 N4Br[M+H] + 365.1341; found 365.1345.

[0110] Embodiment 16:

[0111] Compound 15 was prepared by the following synthetic route, and the specific steps are as follows: take a 25 ml reaction tube, add 3-methyl-5-bromo-o-phenylenediamine (0.1 mmol), tert-butyl isocyanide (0.25 mmol), Pd(PhCN)2Cl2 (10 mol%), carbon tetrabromide (1.0 equiv), silver nitrate (2.0 equiv), pyridine (3.0 equiv) and dimethyl sulfoxide (1 mL) into the reaction tube in sequence, and continue the reaction at 100° C. for 3 hours to obtain the target compound (compound 15, whose structural formula is synthesized by the following technical route), with a yield of 69%.

[0112] The technical route for preparing compound 15 is as follows:

[0113]

[0114] The characterization data of compound 15 are as follows: 1 H NMR (500MHz, DMSO, δppm) 7.34 (d, J = 2.0Hz, 1H), 7.15 (d, J = 1.2Hz, 1H), 6.50 (s, 1H), 6.49 (s, 1H), 2.47 (s, 3H), 1.53 (s, 9H), 1.50 (s, 9H); 13 CNMR(126MHz,DMSO,δppm)143.7,142.6,136.8,135.0,133.6,126.5,125.0,114.9,52.5,52.3,29.1,28.7,17.5.HRMS(ESI)m / z:calcd for C 17 H26 N4Br[M+H] + 365.1341; found 365.1345.

[0115] The beneficial effects of the drug of the present invention are further described below through pharmacodynamic tests and comparative tests. The tumor cell lines used in the experiments of the present invention are drugs for preparing tumors formed by human colon cancer cells HCT116, human liver cancer cells HepG-2, human lung cancer cells A549, human prostate cancer cells PC3, human cervical cancer cells Hela, human glial cancer cells U87, and human malignant breast cancer cells 4T1.

[0116] N 2 ,N 3 - In vitro anti-tumor cell activity test of di-tert-butylquinoxaline-2,3-diamine derivatives:

[0117] 1. Cells and drugs:

[0118] Human colon cancer cells HCT116, human liver cancer cells HepG-2, human lung cancer cells A549, human prostate cancer cells PC3, human cervical cancer cells Hela, human glial cancer cells U87, and human malignant breast cancer cells 4T1 were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences;

[0119] Positive drug: 5-fluorouracil (5-Fu), purchased from Sigma Aldrich Company (St. Louis, MO, USA), purity >99.5% (HPLC).

[0120] 2. Experimental Methods

[0121] Take 5.0×10 tumor cells in the logarithmic growth phase 4 cells / mL were inoculated into 96-well plates, 200 μL per well, and cultured at 37°C and 5% CO2 for 24 h;

[0122] Dissolve the test sample with dimethyl sulfoxide (DMSO), and then use fresh culture medium to prepare a sample culture solution with a final concentration of 1.0-100 μM, add it to a 96-well plate, and incubate it at 37°C and 5% CO2 for 24 hours;

[0123] Remove the culture medium, add 100 μL of 0.5 mg / mL MTT to each well, and incubate at 37°C and 5% CO2 for 3-4 h; carefully remove the culture medium, add 100 μL DMSO, and shake thoroughly to dissolve the crystals;

[0124] The microplate reader uses 570nm as the experimental wavelength and 630nm as the reference wavelength to detect the absorbance value. 50 The values ​​were calculated using the LOGIT method.

[0125] 3. Experimental results

[0126] As shown in Table 1, the experimental results show that N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives have significant inhibitory effects on human colon cancer cells HCT116, human liver cancer cells HepG-2, human lung cancer cells A549, human prostate cancer cells PC3, human cervical cancer cells Hela, human glial cancer cells U87, and human malignant breast cancer cells 4T1. 50 The value is 0.51-67.55μmol / L.

[0127] Table 1N 2 ,N 3 -IC of di-tert-butylquinoxaline-2,3-diamine derivatives on various tumor cells 50 value

[0128]

[0129]

[0130] In summary, the present invention relates to N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives have significant inhibitory effects on colon cancer, liver cancer, lung cancer, prostate cancer, cervical cancer, glioma and breast cancer, and show excellent anti-tumor properties. As a leading anti-tumor drug molecule, the derivative has very important research and development value. Through in-depth research on its mechanism of action, we can further understand its mechanism of action in anti-tumor, thereby providing a more solid theoretical basis for its application in drug research and development. Therefore, the N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives have great development and application prospects in the field of anti-tumor drug research and development. Through further research and optimization, we are expected to develop more efficient and safe anti-tumor drugs, provide new treatment options for cancer patients, and make greater contributions to human health.

[0131] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. Use of a di-tert-butylquinoxaline diamine derivative in the preparation of an anti-tumor pharmaceutical composition, characterized in that: Specifically, N 2 ,N 3 - Application of di-tert-butylquinoxaline-2,3-diamine derivatives in the preparation of anti-tumor drug compositions; The N 2 ,N 3 -The chemical structure of di-tert-butylquinoxaline-2,3-diamine derivative is as follows: Among them, R1 is one of a hydrogen group and a methyl group; R2 is one of a hydrogen group, a fluoro group, a chloro group and a methyl group; R3 is one of a hydrogen group, a methoxy group, a pentyl group, a methyl group, a trifluoromethoxy group, a fluoro group, a chloro group and a bromo group; R4 is one of a hydrogen group and a methyl group.

2. The use according to claim 1, characterized in that: The tumors include, but are not limited to, colon cancer, liver cancer, lung cancer, prostate cancer, cervical cancer, glioma and breast cancer.

3. The use according to claim 1, characterized in that: N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives as candidate drugs or lead compounds for treating tumors.

4. The use according to claim 1, characterized in that: The therapeutic object of the pharmaceutical composition is a mammal or a human.

5. The use according to claim 1, characterized in that: The pharmaceutical composition is N 2 ,N 3 -Di-tert-butylquinoxaline-2,3-diamine derivatives are used as the main component, and auxiliary materials or auxiliary components accepted in pharmacy are added to prepare clinically accepted drug preparations.

6. The use according to claim 1, characterized in that: The N 2 ,N 3 The content of the di-tert-butylquinoxaline-2,3-diamine derivative in the pharmaceutical composition is usually 0.1-95.0% w / w.

7. The use according to claim 5, characterized in that: The pharmaceutical preparations include two dosage forms: oral preparations and injection preparations.

8. The use according to claim 7, characterized in that: The oral preparation is an oral capsule, and the injection preparation is an intravenous injection solution.

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