CN112028831A - Benzopyrazolyl compound, preparation method thereof and application of benzopyrazolyl compound as antitumor drug - Google Patents
Benzopyrazolyl compound, preparation method thereof and application of benzopyrazolyl compound as antitumor drug Download PDFInfo
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Abstract
The invention particularly relates to a benzopyrazolyl compound, a preparation method thereof and application thereof as an anti-tumor drug. The tubulin inhibitor is a novel structural benzopyrazolyl small molecule compound, and the structural general formula is as follows:
experiments further prove that the compound can effectively inhibit the proliferation of human liver cancer cells. The action mechanism is similar to colchicine, and can inhibit tubulin polymerization. The invention has important significance for enhancing the specificity and effectiveness of the medicine, reducing toxic and side effects, preventing drug resistance and the like, and has good potential development and application values.
Description
Technical Field
The invention belongs to the technical field of antitumor active substances, and particularly relates to a benzopyrazolyl compound, a preparation method of the benzopyrazolyl compound and application of the benzopyrazolyl compound as a tubulin inhibitor.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
Microtubules are the main components of the cytoskeleton, are composed of alpha-tubulin and beta-tubulin heterodimers, have the dynamic characteristics of polymerization and depolymerization, and play an important role in the processes of maintaining cell morphology, mitosis, signal transduction, substance transportation and the like. Since cancer cells are largely distinguished from normal cells by their uncontrolled, abnormally active, rapid division, and microtubules play an extremely important role in the M phase, they not only participate in the formation of spindles, but also their kinetic velocity in the M phase can be increased by 20-100 times. Therefore, the tubulin inhibitor is combined with a specific site on tubulin to influence and interfere the polymerization and depolymerization kinetics of the tubulin, inhibit the activity of microtubules in the M phase of tumor cells, influence the mitotic process of the tumor cells, block the formation of spindle bodies in the M phase, and cause mitotic failure or arrest, thereby inhibiting the growth and proliferation of the tumor cells, leading to the apoptosis of microtubule-resistant drugs to become a main chemotherapeutic drug, and being widely used for the clinical treatment of various tumors. The tubulin inhibitor influences and interferes with the polymerization and depolymerization kinetics of tubulin by binding with a specific site on the tubulin, thereby blocking the formation of spindle in M phase and arresting the growth of tumor cells in G2/M phase. Currently, clinically used microtubule inhibitors mainly include drugs for inhibiting tubulin depolymerization, represented by paclitaxel, and drugs for inhibiting microtubule polymerization, represented by vinblastine. However, the drugs have the problems of great toxic and side effects, easy generation of drug resistance, complex structure, great synthesis difficulty and the like, so that the search for a novel, efficient and low-toxicity microtubule inhibitor becomes a hotspot of the research of the current antitumor drugs.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides a compound with a structure shown in a formula I, and a preparation method and application thereof. The compound has the function of inhibiting the polymerization activity of tubulin, is a novel small molecular tubulin inhibitor, and can effectively inhibit the proliferation of human liver cancer cells through further verification by experiments. The invention has important significance for enhancing the specificity and the effectiveness of the medicine, reducing the toxic and side effects, preventing the drug resistance and the like.
Based on the technical effects, the invention provides the following technical scheme:
in a first aspect of the present invention, there is provided a compound selected from compounds of formula i, or pharmaceutically acceptable salts, solvates and hydrates thereof, said compound having the structure:
wherein R is H, 4-fluorine, 4-methoxy or 2-methoxy.
The compound with the structure has an inhibition effect on tumor cells, and further research also confirms that the compound has an inhibition effect on the proliferation of the tumor cells as a tubulin inhibitor. The compound belongs to a small molecular entity, is applied to research and development of antitumor drugs, is expected to obtain better solubility and membrane permeation effect, and improves bioavailability. In addition, the invention also provides a preparation method of the compound, based on the preparation method provided by the invention, most reaction steps can be carried out based on normal temperature, no toxic reagent is introduced in the reaction process, high temperature and high pressure are not needed, the preparation process is safer, and the preparation method is very friendly to industrial expanded production.
In a second aspect of the present invention, a preparation method of the compound of the first aspect is provided, where the preparation method includes using the compound of formula 1 as a starting material to construct a benzopyrazole core structure, so as to obtain a compound of formula 2; the compound shown in the formula 2 is subjected to acetylation to obtain a compound shown in a formula 3, then the compound is reacted with phenylboronic acid or substituted phenylboronic acid to obtain a compound shown in a formula 4, and finally, deacetylation is performed to obtain a compound shown in a formula I, wherein the reaction flow is shown as the following formula:
in a third aspect of the invention, there is provided the use of a compound of formula I as a tubulin inhibitor.
The research of the invention finds that when the R substituent is 4-F or 4-MeO, the inhibitor has obvious inhibition effect on liver cancer cells, and the inhibition effect is improved by five to six times compared with the inhibition effect of other two substituents.
In addition, the invention verifies that the compound has tubulin inhibition effect, and based on the effect, the compound shown in the formula I is expected to be applied to the development of antitumor drugs which are possibly taken as main active substances; since the art also employs a combination of a number of different mechanisms of antineoplastic drug combination therapy strategies, the present invention also provides regimens for the use of the compounds in combination with other active ingredients.
Accordingly, in a fourth aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of formula i in the first aspect.
In a fifth aspect, an anti-tumor drug is provided, which comprises the pharmaceutical composition of the fourth aspect.
The beneficial effects of one or more technical schemes are as follows:
(1) the benzopyrazoles small molecule tubulin inhibitor designed by the invention has a novel structure and has important significance for enhancing the specificity and effectiveness of the medicament, reducing toxic and side effects, preventing drug resistance and the like.
(2) The compound is a novel benzopyrazolyl small molecule tubulin inhibitor, has strong proliferation inhibition activity on human liver cancer HepG2 cells, can inhibit tubulin polymerization, and has good potential development and application values.
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The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.
FIG. 1 shows the results of the in vitro inhibition of tubulin polymerization experiments with the compounds described in examples 1-4.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
As introduced in the background art, aiming at the defects of high synthesis difficulty, high side effect and multidrug resistance of the prior tubulin inhibitor, the invention provides a benzopyrazolyl compound which can be used as the tubulin inhibitor in order to solve the technical problems.
In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.
In a first aspect of the present invention, there is provided a compound selected from compounds of formula i, or pharmaceutically acceptable salts, solvates and hydrates thereof, said compound having the structure:
wherein R is H, 4-fluorine, 4-methoxy or 2-methoxy.
In a second aspect of the present invention, a preparation method of the compound of the first aspect is provided, where the preparation method includes using the compound of formula 1 as a starting material to construct a benzopyrazole core structure, so as to obtain a compound of formula 2; the compound shown in the formula 2 is subjected to acetylation to obtain a compound shown in a formula 3, then the compound is reacted with phenylboronic acid or substituted phenylboronic acid to obtain a compound shown in a formula 4, and finally, deacetylation is performed to obtain a compound shown in a formula I, wherein the reaction flow is shown as the following formula:
preferably, the compound of formula 2 is prepared as follows: dissolving the compound shown in the formula 1 and p-nitrobromobenzene in an organic solvent, adding a reducing agent in batches, stirring at room temperature for reaction, slowly adding the reaction liquid into cold water after the reaction is finished, and filtering to obtain a yellow solid part, namely the compound shown in the formula 2.
Further preferably, the organic solvent includes but is not limited to DMF.
Further preferably, the reaction time at room temperature is 2.5-3.5 h.
Further preferably, the reducing agent is sodium hydride or sodium borohydride.
In some specific embodiments of the above preferred embodiments, the compound of formula 2 is prepared as follows: dissolving the compound shown in the formula 1 and p-nitrobromobenzene in DMF, stirring and reacting for 3h at room temperature, slowly adding the reaction liquid into cold water after the reaction is finished, washing the precipitated yellow solid by distilled water, and drying to obtain a yellow solid, namely the compound shown in the formula 2.
Preferably, the compound represented by formula 3 is prepared as follows: and (3) mixing the compound shown in the formula 2 with acetyl chloride and triethylamine, stirring at normal temperature for reaction, adding water for quenching after the reaction is completed, adding dichloromethane into a reaction system for extraction, and performing column chromatography purification on a dichloromethane extraction part to obtain the compound shown in the formula 3.
More preferably, the compound represented by formula 2 is mixed with acetyl chloride and triethylamine, and then dissolved in dichloromethane and stirred at normal temperature.
Further preferably, the reaction time is 9-12 h.
Further preferably, the column chromatography is dry silica gel column chromatography.
In some specific embodiments of the above preferred embodiments, the compound of formula 3 is prepared by the following steps: dissolving the compound shown in the formula 2 in dichloromethane, adding acetyl chloride and triethylamine, and stirring at room temperature for reaction for 10 hours; adding water to quench the reaction, extracting with dichloromethane, and purifying by dry silica gel column chromatography to obtain the compound of formula 3.
Preferably, the compound represented by formula 4 is prepared as follows: dissolving the compound shown in formula 3 in dioxane and water to obtain a mixed solvent, and adding phenylboronic acid or substituted phenylboronic acid, Pd (PPh)3)4Heating potassium carbonate for reaction; and extracting with ethyl acetate, and purifying the ethyl acetate extraction part by column chromatography to obtain the compound shown in the formula 4.
Further preferably, the substituted phenylboronic acids include, but are not limited to, 4-fluorophenylboronic acid, 4-methoxyphenylboronic acid, and 2-methoxyphenylboronic acid.
Further preferably, the heating reaction is heating reflux, and the reaction time is 2.5-3.5 h.
Further preferably, the column chromatography is silica gel column chromatography.
Preferably, the compound of formula I is prepared as follows: dissolving the compound of the formula 4 in methanol, adding LiOH in batches, and stirring at room temperature for reaction; after the reaction is completed, water is added to obtain a solid part, namely the compound shown in the formula I.
Further preferably, the room temperature reaction time is 0.5-1.5 h.
In some embodiments of the above preferred embodiments, the compound is prepared by the following formula:
in a third aspect of the invention, there is provided the use of a compound of formula I as a tubulin inhibitor.
Preferably, in the compound shown in the formula I, R is 4-F or 4-MeO. The research of the invention finds that when the R substituent is 4-F or 4-MeO, the inhibitor has obvious inhibition effect on liver cancer cells, and the inhibition effect is improved by five to six times compared with the inhibition effect of other two substituents.
In a fourth aspect of the invention, there is provided a pharmaceutical composition comprising a compound of formula i in the first aspect.
Preferably, the pharmaceutical composition further comprises other antitumor active ingredients or auxiliary therapeutic ingredients.
Further preferably, the antitumor active ingredients include but are not limited to alkylating agents and nitrogen mustards (such as nitrogen mustards, chlorambucil, cyclophosphamide, ifosfamide, melphalan and the like), thiotepa (such as thiotepa and the like), nitrosoureas (such as carmustine, semustine and the like) and methylsulfonates (such as busulfan and the like); platinum compounds (such as cisplatin, carboplatin, platinum oxalate and the like) and mitomycin (such as mitomycin and the like); dihydrofolate reductase inhibitors (e.g., methotrexate, pemetrexed, etc.), thymidine synthase inhibitors (e.g., 5-FU, FT-207, capecitabine, etc.), purine nucleotide synthase inhibitors (e.g., 6-mercaptopurine, 6-TG, etc.), ribonucleotide reductase inhibitors (e.g., hydroxyurea, etc.), and DNA polymerase inhibitors (e.g., cytarabine, gemcitabine, etc.); nucleic acid transcription inhibitors (e.g., actinomycin D, daunorubicin, doxorubicin, epirubicin, aclarubicin, mithramycin, etc.), topoisomerase inhibitors (e.g., irinotecan, topotecan, hydroxycamptothecin, etc.), tubulin inhibitors (e.g., paclitaxel, docetaxel, vinblastine, catharanthine, podophylline, homoharringtonine, etc.), other cytotoxic agents (e.g., asparaginase); hormone drugs, biological response modifiers, monoclonal antibodies, and the like, and also other drugs which are proved to have an inhibitory effect on tumors.
Further preferably, the adjuvant therapy component includes, but is not limited to, a blood-increasing agent (e.g., G-CSF, GM-CSF, interleukin-11, EPO, etc.), an antiemetic agent (e.g., ondansetron, granisetron hydrochloride, etc.), an analgesic agent (e.g., aspirin, paracetamol, codeine, tramadol, morphine, fentanyl, etc.), an osteoclast-inhibiting agent (e.g., disodium clodronate, disodium pamidronate, etc.).
In a fifth aspect of the present invention, an anti-tumor drug is provided, wherein the anti-tumor drug comprises the pharmaceutical composition of the fourth aspect.
Preferably, the anti-tumor drug is in a dosage form including, but not limited to, oral preparations, injections, aerosols and external preparations.
Further preferably, the oral preparation is one of tablets, capsules, oral liquid dosage forms, oral pills, oral granules and oral powders.
Further preferably, the injection includes but is not limited to freeze-dried powder injection, emulsion, suspension and other liquid preparations.
The following example relates to the compound of formula 1, p-toluenesulfonyl (3,4, 5-methoxybenzaldehyde) hydrazone, which is a commercially available product, CAS: 188493-61-0.
Example 1 preparation of compound a1 (formula I, R ═ H)
(1) P-toluenesulfonyl (3,4, 5-methoxybenzaldehyde) hydrazone (formula 1) (2.0g, 5.48mmol) and p-nitrobenzyl bromide (1.22g,6.03mmol) were dissolved in DMF (20mL), NaH (0.66g, 16.44mmol) was added in portions, and the reaction was stirred at room temperature for 3 h. The reaction was slowly added to about 200mL of cold water to precipitate a yellow solid. And (4) carrying out suction filtration, washing with distilled water, and drying a filter cake to obtain a compound shown in the formula 2, wherein 2.05g of a yellow solid is obtained.1H NMR(600MHz,CDCl3)8.11(d,J＝1.8Hz,1H),7.50(dd,J＝9.0,1.8Hz,1H),7.36(d,J＝9.0Hz,1H),7.12(s,2H),3.96(s,6H),3.93(s,3H).
(2) The compound of formula 2 (174mg, 0.48mmol) was dissolved in dichloromethane (6mL), acetyl chloride (113mg, 1.44mmol) and triethylamine (290mg, 2.87mmol) were added, and the reaction was stirred at room temperature for 10 h. The reaction was quenched with water, extracted with dichloromethane and purified by dry silica gel column chromatography to give the compound of formula 3 as a yellow solid (107 mg).1H NMR(600MHz,CDCl3)8.41(d,J＝8.8Hz,1H),8.06(d,J＝1.8Hz,1H),7.69(dd,J＝8.8,1.8Hz,1H),7.10(s,2H),3.98(s,6H),3.94(s,3H),2.84(s,3H).
(3) The compound of formula 3 (200mg, 0.49mmol) was dissolved inA mixed solvent of dioxane (10mL) and water (0.4mL) was added phenylboronic acid (90mg,0.74mmol), Pd (PPh3)4(33mg), and potassium carbonate (205mg, 1.48mmol), and the reaction was refluxed for 3 hours. The reaction was quenched with water, extracted with ethyl acetate, and purified by silica gel column chromatography to give the compound of formula 4 (R ═ H) as a white solid (44 mg).1H NMR(600MHz,CDCl3)8.57(dd,J＝9.0,0.6Hz,1H),8.09(dd,J＝1.8,0.6Hz,1H),7.84(dd,J＝9.0,1.8Hz,1H),7.66–7.62(m,2H),7.51–7.46(m,2H),7.39(tt,J＝7.2,1.8Hz,1H),7.19(s,2H),3.98(s,6H),3.94(s,3H),2.87(s,3H).
(4) The compound of formula 4 (R ═ H) (44mg,0.11mmol) was dissolved in methanol (6mL), and LiOH · H was added2O (46mg,1.10mmol), the reaction was stirred at room temperature for 1 h. Water was added to precipitate a white solid. Suction filtration and drying of the filter cake gave compound a1, a compound of formula I (R ═ H), as a yellow solid 39 mg.1H NMR(600MHz,CDCl3)8.16(d,J＝1.2Hz,1H),7.71(dd,J＝8.4,1.2Hz,1H),7.64(dd,J＝8.4,1.2Hz,2H),7.58–7.55(m,1H),7.50–7.46(m,2H),7.37(tt,J＝7.2,1.2Hz,1H),7.22(s,2H),3.97(s,6H),3.94(s,3H).
Example 2: preparation of compound a2 (formula I, R ═ 4-F)
(1) The compound of formula 3 (94mg, 0.23mmol) was dissolved in a mixed solvent of dioxane (5mL) and water (0.2mL), and 4-fluorobenzeneboronic acid (39mg, 0.28mmol), Pd (PPh3)4(16mg), potassium carbonate (96mg) were added to conduct a reaction at reflux for 14 h. The reaction was quenched with water, extracted with ethyl acetate, and purified by silica gel column chromatography to give the compound of formula 4 (R ═ 4-F) as a yellow solid (79.2 mg).1H NMR(600MHz,CDCl3)8.57(d,J＝9.0Hz,1H),8.03(d,J＝1.8Hz,1H),7.78(dd,J＝9.0,1.8Hz,1H),7.61–7.55(m,2H),7.20–7.14(m,4H),3.98(s,6H),3.94(s,3H),2.87(s,3H).
(2) The compound of formula 4 (R ═ 4-F) (52mg,0.12mmol) was dissolved in methanol (5mL), and LiOH · H was added2O (52mg,1.24mmol), the reaction was stirred at room temperature for 1 h. Water was added to precipitate a white solid. Filtering, drying the filter cake to obtain compound A2, i.e. the compound of formula I (R is 4-F) with yellow color36mg of solid.1H NMR(600MHz,CDCl3)10.32(brs,1H),8.09(s,1H),7.63(dd,J＝8.4,1.2Hz,1H),7.60–7.55(m,3H),7.20(s,2H),7.18–7.14(m,2H),3.96(s,6H),3.93(s,3H).
Example 3: preparation of compound a3 (formula I, R ═ 4-MeO)
(1) The compound of formula 3 (150mg, 0.37mmol) was dissolved in a mixed solvent of dioxane (7.5mL) and water (0.3mL), 4-methoxyphenylboronic acid (68mg, 0.44mmol), Pd (PPh3)4(17mg), and potassium carbonate (153mg) were added, and the reaction was refluxed for 12 hours. The reaction was quenched with water, extracted with ethyl acetate, and purified by silica gel column chromatography to give the compound of formula 4 (R ═ 4-methoxy) as a white solid (50 mg).1H NMR(600MHz,CDCl3)8.54(dd,J＝9.0,0.6Hz,1H),8.03(d,J＝1.8Hz,1H),7.80(dd,J＝9.0,1.8Hz,1H),7.57(d,J＝9.0Hz,2H),7.19(s,2H),7.02(d,J＝9.0Hz,2H),3.98(s,6H),3.94(s,3H),3.87(s,3H),2.87(s,3H).
(2) The compound of formula 4 (R ═ 4-methoxy) (30mg,0.07mmol) was dissolved in methanol (3mL), and LiOH · H was added2O (29mg,0.70mmol), the reaction was stirred at room temperature for 1.5 h. Water was added to precipitate a white solid. Suction filtration and filter cake drying gave compound a3, a compound of formula I (R ═ 4-methoxy), as a yellow solid, 24 mg.1H NMR(600MHz,CDCl3)8.10(s,1H),7.66(dd,J＝8.4,1.8Hz,1H),7.56(d,J＝9.0Hz,2H),7.52(d,J＝8.4Hz,1H),7.21(s,2H),7.01(d,J＝9.0Hz,2H),3.96(s,6H),3.94(s,3H),3.87(s,3H).
Example 4: preparation of compound a4 (formula I, R ═ 2-MeO)
(1) The compound of formula 3 (150mg, 0.37mmol) was dissolved in a mixed solvent of dioxane (7.5mL) and water (0.3mL), and 2-methoxyphenylboronic acid (68mg, 0.44mmol), Pd (PPh3)4(17mg), potassium carbonate (153mg) were added to conduct a reflux reaction for 15 h. Adding water to quench the reaction, and adding ethyl acetateAfter the ester extraction, the product was purified by silica gel column chromatography to give the compound of formula 4 (R ═ 2-methoxy) as an off-white solid (81 mg).1H NMR(600MHz,CDCl3)8.54(d,J＝8.4Hz,1H),8.08(s,1H),7.78(dd,J＝8.4,1.8Hz,1H),7.36(m,2H),7.20(s,2H),7.07(td,J＝7.2,1.2Hz,1H),7.05–7.02(m,1H),3.96(s,6H),3.93(s,3H),3.83(s,3H),2.87(s,3H).
(2) The compound of formula 4 (R ═ 2-methoxy) (61mg,0.14mmol) was dissolved in methanol (5mL), and LiOH · H2O (59mg,1.40mmol) was added, and the reaction was stirred at room temperature for 1.5H. Water was added to precipitate a white solid. Suction filtration and filter cake drying gave compound a4, a compound of formula I (R ═ 2-methoxy), as a white solid, 50 mg.1H NMR(600MHz,CDCl3)10.13(brs,1H),8.12(s,1H),7.64(d,J＝8.4Hz,1H),7.54(d,J＝8.4Hz,1H),7.41–7.33(m,2H),7.21(s,2H),7.09–7.00(m,2H),3.95(s,6H),3.92(s,3H),3.84(s,3H).
Example 5: antitumor cell proliferation assay
1. The experimental method comprises the following steps:
human hepatoma HepG2 cells were dosed with the compound of formula I at different concentrations and placed at 37 ℃ with 5% CO2The culture box is incubated for 72 hours, and the inhibition rate of the compound on tumor cells is measured by a tetramethyl azoazolate (MTT) colorimetric method, and the result is shown in table 1.
2. And (3) test results:
TABLE 1 half inhibitory concentration of the compounds of examples 1-4 on HepG2 cells from liver cancer
aMean ± Standard Deviation (SD) of three experiments are shown.
As can be seen from the data in Table 1, the four compounds all have inhibitory effects on hepatoma cells, the effects of A2 and A3 are particularly prominent, and the inhibitory effects of the compounds are remarkably improved compared with those of the compounds A1 and A4.
Example 6: tubulin polymerization assay
1. Experimental drugs: the compound prepared in example 2 (noted as a2), colchicine and dimethyl sulfoxide (DMSO).
2. The experimental method comprises the following steps:
tubulin Polymerization experiments were performed according to the Tubulin Polymerization Assay Kit instructions from Cytoskeleton.
3. The results of the experiment are shown in FIG. 1. As can be seen from FIG. 1, Compound A2 prepared in example 2 inhibited tubulin polymerization, IC50It was 2.22. mu.M.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A compound selected from compounds of formula i, or pharmaceutically acceptable salts, solvates, and hydrates thereof, said compound of formula i having the structure:
wherein R is H, 4-fluorine, 4-methoxy or 2-methoxy.
2. The preparation method of the compound as claimed in claim 1, which comprises the steps of taking the compound as shown in formula 1 as a starting material, constructing a benzopyrazole mother nucleus structure to obtain the compound as shown in formula 2; the compound shown in the formula 2 is subjected to acetylation to obtain a compound shown in a formula 3, then the compound is reacted with phenylboronic acid or substituted phenylboronic acid to obtain a compound shown in a formula 4, and finally, deacetylation is performed to obtain a compound shown in a formula I, wherein the reaction flow is shown as the following formula:
3. a process for preparing a compound of claim 2, wherein the compound of formula 2 is prepared by: dissolving a compound shown as a formula 1 and p-nitrobromobenzene in an organic solvent, adding a reducing agent in batches, stirring at room temperature for reaction, slowly adding a reaction solution into cold water after the reaction is finished, and filtering to obtain a yellow solid part, namely a compound shown as a formula 2;
preferably, the organic solvent is a solvent including, but not limited to DMF;
preferably, the reaction time at room temperature is 2.5-3.5 h;
preferably, the reducing agent is sodium hydride or sodium borohydride;
further, the compound represented by formula 2 is prepared as follows: dissolving the compound shown in the formula 1 and p-nitrobromobenzene in DMF, stirring and reacting for 3h at room temperature, slowly adding the reaction liquid into cold water after the reaction is finished, washing the precipitated yellow solid by distilled water, and drying to obtain a yellow solid, namely the compound shown in the formula 2.
4. The method of preparing the compound of claim 2, wherein the compound of formula 3 is prepared as follows: mixing the compound shown in the formula 2, acetyl chloride and triethylamine, stirring at normal temperature for reaction, adding water for quenching after the reaction is completed, adding dichloromethane into a reaction system for extraction, and performing column chromatography purification on a dichloromethane extraction part to obtain the compound shown in the formula 3;
preferably, the compound shown in the formula 2, acetyl chloride and triethylamine are mixed and then dissolved in dichloromethane to be stirred at normal temperature;
preferably, the reaction time is 9-12 h;
preferably, the column chromatography is dry silica gel column chromatography;
further preferably, the specific preparation method of the compound represented by the formula 3 is as follows: dissolving the compound shown in the formula 2 in dichloromethane, adding acetyl chloride and triethylamine, and stirring at room temperature for reaction for 10 hours; adding water to quench the reaction, extracting with dichloromethane, and purifying by dry silica gel column chromatography to obtain the compound of formula 3.
5. The method of preparing the compound of claim 2, wherein the compound of formula 4 is prepared as follows: dissolving the compound shown in formula 3 in dioxane and water to obtain a mixed solvent, and adding phenylboronic acid or substituted phenylboronic acid, Pd (PPh)3)4Heating potassium carbonate for reaction; extracting with ethyl acetate, and purifying the ethyl acetate extract by column chromatography to obtain a compound of formula 4;
preferably, the substituted phenylboronic acids include, but are not limited to, 4-fluorophenylboronic acid, 4-methoxyphenylboronic acid, and 2-methoxyphenylboronic acid;
preferably, the heating reaction is heating reflux, and the reaction time is 2.5-3.5 h;
preferably, the column chromatography is purified by silica gel column chromatography.
6. A process for the preparation of a compound according to claim 2, wherein the compound of formula i is prepared as follows: dissolving the compound of the formula 4 in methanol, adding LiOH in batches, and stirring at room temperature for reaction; adding water after the reaction is completed to obtain a solid part, namely the compound shown in the formula I; preferably, the room-temperature reaction time is 0.5-1.5 h.
7. A process for the preparation of a compound according to any one of claims 3 to 6, wherein the compound is prepared according to the formula:
8. the use of a compound of formula i as defined in claim 1 as a tubulin inhibitor;
preferably, in the compound shown in the formula I, R is 4-F or 4-MeO.
9. A pharmaceutical composition comprising a compound of formula i of claim 1;
preferably, the pharmaceutical composition also comprises other antitumor active ingredients or auxiliary therapeutic ingredients;
further preferred, the anti-tumor active ingredients include, but are not limited to, alkylating agents and nitrogen mustards, thiotepas, nitrosoureas and methyl sulfonates; platinum compounds and mitomycin; dihydrofolate reductase inhibitors, thymidine synthase inhibitors, purine nucleotide synthase inhibitors, ribonucleotide reductase inhibitors and DNA polymerase inhibitors; nucleic acid transcription inhibitors, topoisomerase inhibitors, tubulin inhibitors, other cytotoxic agents; hormone drugs, biological response modifiers, monoclonal antibodies and the like, and also other drugs which are proved to have an inhibiting effect on tumors;
further preferably, the adjuvant therapy component includes, but is not limited to, a blood-increasing agent, an antiemetic agent, an analgesic agent, an osteoclast-inhibiting agent.
10. An antitumor agent comprising the pharmaceutical composition according to claim 9;
preferably, the anti-tumor drug is in a dosage form including but not limited to oral preparations, injections, aerosols and external preparations;
further preferably, the oral preparation is one of tablets, capsules, oral liquid dosage forms, oral pills, oral granules and oral powder;
further preferably, the injection includes but is not limited to freeze-dried powder injection, emulsion, suspension and other liquid preparations.
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Cited By (1)
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| CN116514780A (en) * | 2023-04-26 | 2023-08-01 | 山东第一医科大学附属省立医院(山东省立医院) | Indazole micromolecular tubulin inhibitor and preparation method and application thereof |
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