CN110981870B - Beta-carboline-cycloenone derivative based on dual responses of pH and GSH and application thereof - Google Patents

Beta-carboline-cycloenone derivative based on dual responses of pH and GSH and application thereof Download PDF

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CN110981870B
CN110981870B CN201911256603.7A CN201911256603A CN110981870B CN 110981870 B CN110981870 B CN 110981870B CN 201911256603 A CN201911256603 A CN 201911256603A CN 110981870 B CN110981870 B CN 110981870B
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pyrido
indol
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oxocyclohex
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凌勇
刘季
刘昕
凌长春
张延安
钱建强
孟迟
杭佳颖
陈苏蒙
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Abstract

The invention discloses a beta-carboline-cycloenone derivative, which has a structure shown in the following general formula:

Description

Beta-carboline-cycloenone derivative based on dual responses of pH and GSH and application thereof
Technical Field
The invention relates to the field of biological medicines, in particular to a beta-carboline-cycloenone derivative based on dual responses of pH and GSH, a preparation method thereof and medical application of a medicine for inhibiting tumor proliferation by targeting GSH/GST pi, and especially application of the beta-carboline-cycloenone derivative in preparing an anti-tumor diagnosis and treatment agent.
Background
Malignant tumor seriously threatens human health and life, the number of cancer-caused deaths is rapidly increased, and the malignant tumor becomes the first death disease in the world and becomes a global challenge and problem. Research and development of diagnostic therapeutic agents, which combine diagnosis and treatment into one, and immediately administer effective treatment (surgery and/or drug) while making diagnosis, have now become one of the research hotspots in the medical field, improving the efficiency of treatment and specificity of drug release. The main components are small molecular diagnosis and treatment agent and macromolecular nano diagnosis and treatment agent. The former is easier to prepare, and usually employs a prodrug strategy, in which an anticancer drug, an imaging agent, and an activation unit are linked by covalent bonds. The small molecule diagnosis and treatment agent generally has better fluorescence imaging capability, can be induced by tumor cell related molecules to synergistically release drugs, and improves the selectivity of the small molecule diagnosis and treatment agent on solid tumors, thereby improving the anticancer effect. Compared with the macromolecule nano diagnosis and treatment agent, the small molecule prodrug system has better biocompatibility and cellular absorbability and can be regenerated.
The Tumor Microenvironment (TME) has unique physicochemical properties, is obviously different from normal tissues, and refers to a special environment for the growth of tumor cells formed after the interaction of the tumor cells and extracellular matrixes in the growth process of the tumor cells. Early dynamic tracking of the tumor and its microenvironment will facilitate its surgical resection and/or precise drug treatment. TME not only provides conditions for the growth of tumor cells, but also is a necessary site for tumor cell metastasis. Given that tumor cells and their microenvironment undergo glycolysis under hypoxic conditions, H is excreted+The tumor tissue has obvious slight acidity (pH is 6.5-6.8), and the normal tissue has pH of 7.2-7.4; furthermore, organelles with higher acidity, such as lysosomes (pH 4.5-5.0), are also present in tumor cells; therefore, the research on the pH-sensitive fluorescent probe has important significance in selectively targeting the tumor cells and the microenvironment thereof. However, the conventional pH-sensitive fluorescent probe is mainly activated by an acid-sensitive hydrazone bond or an acetal fragment, but the groups themselves have problems of instability in vivo, slow color development, easy metabolism, and the like.
Glutathione mercaptotransferases, glutathione-S-transferases (GSTs), are a multifunctional isozyme widely present in the body of mammals. The main function of the method is to catalyze the sulfhydryl of Glutathione (GSH) to carry out nucleophilic attack on endogenous or exogenous electrophilic groups (such as carbon, nitrogen, sulfur and the like) to carry out coupling reaction, so as to form a metabolic product with higher water solubility, and the metabolic product is easy to excrete from bile or urine. The structure of human cellular GSTs is closely related to the development and progression of human diseases. Human cytoplasmic GSTs can be divided into 7 subtypes, according to differences in amino acid sequence, physical structure and immunological cross-reactivity, which are: alpha, mu, pi, sigma, theta, omega and zeta, wherein GST pi has close relationship with human tissue cell canceration, tumor formation and generation of tumor resistance. Studies have shown that GST pi is overexpressed in a variety of tumor cells (e.g., breast cancer, colon cancer tumor cells) as well as in resistant tumors. In particular, in liver and intestinal tissues, the formation of GST pi is considered to be one of the markers of liver cancer formation. The characteristic of GST pi makes it an important target in designing antitumor drug prodrug. Therefore, the development and design of cancer chemotherapy drugs and probes by utilizing the highly expressed GSTpi of tumor tissues have great application value.
Disclosure of Invention
The invention carries out structural modification on the basis of natural indole alkaloid beta-carboline with anti-tumor activity, connects the beta-carboline with cycloalkenone through carbamate bond, introduces an electron-donating group on the structure of the beta-carboline, can generate pH sensitive fluorescence, can release fluorescence at 492nm after being covalently combined with GSH, simultaneously has targeting GSTpi, obviously enhances the anti-tumor proliferation and metastasis activity of the compound, and achieves the dual targeting effect of early diagnosis and accurate treatment of cancer.
The specific technical scheme of the invention is as follows: the beta-carboline-cycloenone derivative has a structure shown in the following general formula:
Figure GDA0002639606720000021
wherein,
R1or R2The same or different, represents 1 or more substituents on the corresponding substituted ring, and is selected from one or more of H, amino, halogen, hydroxyl, nitro, alkoxy, alkyl and alkylamino, R1Or R2When each represents a plurality of substituents,each substituent is the same or different;
R3selected from H, benzyl, allyl, alkyl, methoxyalkyl;
R4selected from H, alkyl, methoxyalkyl or
Figure GDA0002639606720000031
n is 1, 2 or 3.
Preferably, R1Or R2One or more selected from H, amino, halogen, hydroxyl, nitro, alkoxy of C1-C6, alkyl of C1-C6 and alkylamino of C1-C6, and more preferably one or more selected from H, F, Cl, Br, I, hydroxyl, amino, nitro, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, methylamino, ethylamino, methylethylamino and N, N-dimethylamino;
R3selected from H, benzyl, allyl, C1-C6 alkyl, C1-C6 methoxyalkyl, more preferably H, benzyl, allyl, methyl, ethyl, propyl, isopropyl, methoxymethyl, methoxyethyl, methoxypropyl or methoxyisopropyl;
R4selected from H, C1-C6 alkyl, C1-C6 methoxyalkyl or
Figure GDA0002639606720000032
More preferably H, methyl, ethyl, propyl, isopropyl, methoxymethyl, methoxyethyl, methoxypropyl, methoxyisopropyl or
Figure GDA0002639606720000033
n is 1, 2 or 3.
The invention relates to a specific beta-carboline/cyclic ketene derivative, wherein R is1 Represents 3,4,5-Tri-OCH3、4-CH3、3-OCH3、4-OCH3、4-N,N-Di-CH3、2,4-Di-OCH3、2,5-Di-OCH3、3,4-Di-OCH3Or 3,5-Di-OCH3；R2Or R3Represents H; r4Represents H or
Figure GDA0002639606720000034
n is 1, 2 or 3.
The beta-carboline/cycloalkenone derivatives related to the embodiments of the present invention have the following structures:
TABLE 1
Figure GDA0002639606720000041
Ⅰ1: (6-Oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ1: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ2: (6-Oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ2: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ3: (6-Oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ3: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ4: (6-Oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ4: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ5: (6-Oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ5: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ6: (6-Oxocyclohex-1-en-1-yl) methyl (1- (2, 4-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ6: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ7: (6-Oxocyclohex-1-en-1-yl) methyl (1- (2, 5-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ7: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ8: (6-Oxocyclohex-1-en-1-yl) methyl (1- (3, 4-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ8: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ9: (6-Oxocyclohex-1-en-1-yl) methyl (1- (3, 5-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ9: bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ10: (6-Oxocyclopent-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ10: bis (6-oxocyclopent-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ11: (6-Oxocyclopent-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ11: bis (6-oxocyclopent-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ12: (6-Oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ12: bis (6-oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅰ13: (6-Oxocyclohept-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamate;
Ⅱ13: bis (6-oxocyclohept-1-en-1-yl) methyl (1- (4-N, N dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl) carbamates.
Another object of the present invention is to provide a process for the preparation of a compound of the formula of the present invention, comprising the steps of:
(1) reacting compound 1 in hydrazine hydrate to obtain compound 2, preferably reacting with 85% hydrazine hydrate in methanol,
Figure GDA0002639606720000061
(2) reacting compound 2 with NaNO2The reaction is carried out under acidic conditions, preferably diluted hydrochloric acid conditions, to obtain a compound 3,
Figure GDA0002639606720000071
(3) reacting the compound 3 under an acidic condition, preferably an acetic acid aqueous solution to obtain a compound 4;
Figure GDA0002639606720000072
(4) reacting compound 5 with p-nitrophenyl chloroformate under basic conditions, preferably DIPEA conditions, to give compound 6,
Figure GDA0002639606720000073
(5) reacting compound 4 with compound 6 under basic conditions, preferably DIPEA conditions, to give compound 7 and/or compound 8,
Figure GDA0002639606720000081
or, the synthesis step also comprises the step (6), the compound 7 is reacted under sodium hydrogen, and then reacted with benzyl bromide, allyl bromide or alkyl bromide to obtain the compound 9,
Figure GDA0002639606720000082
R1or R2The same or different, represents 1 or more substituents on the corresponding substituted ring, and is selected from one or more of H, amino, halogen, hydroxyl, nitro, alkoxy, alkyl and alkylamino, R1Or R2When each represents a plurality of substituents, each substituent may be the same or different;
R3selected from H, benzyl, allyl, alkyl, methoxyalkyl;
R4selected from H, alkyl or methoxyalkyl;
n is 1, 2 or 3.
The invention also aims to provide application of the beta-carboline-cycloalkenone derivative in preparing a targeting GST pi drug or probe. On one hand, after the beta-carboline/cycloalkenone derivative is targeted to GST pi highly expressed by tumor tissues, the cycloalkenone segment in the structure of the compound can be specifically identified with the sulfhydryl of GSH and generates Michael addition, and GSH response fluorescence is generated. The compound of the invention has pH sensitivity fluorescence, and when the pH is reduced from 8 to 4, the fluorescence is obviously enhanced. On the other hand, after the compound beta-carboline-cycloenone derivative targets GST pi highly expressed by tumor tissues, the compound beta-carboline-cycloenone derivative can be selectively activated by GSH/GST pi in tumor cells to release pharmacological activity segments 3-glutathione-2-exomethylene cyclic ketone and beta-carboline segments, and the pharmacological activity effects such as anti-tumor proliferation and transfer activity are obviously enhanced. The drug capable of targeting GST pi is a drug for treating and/or preventing cancer, preferably, the cancer is liver cancer, colon cancer, cervical cancer or gastric cancer.
The compound has better fluorescence imaging capability and high selectivity on solid tumors, and can achieve the dual purposes of diagnosis and treatment.
The compounds of the invention can be formulated for administration either alone or in combination with one or more pharmaceutically acceptable carriers. For example, solvents, diluents, etc., and can be used in oral administration forms such as capsules, dispersible powders, tablets, granules, etc. The various dosage forms of the pharmaceutical compositions of the present invention may be prepared according to methods well known in the pharmaceutical art. Such pharmaceutical formulations may contain, for example, from 0.05% to 90% by weight of the active ingredient, more usually between about 15% and 60% by weight of the active ingredient, in combination with a carrier. The dosage of the compound can be 0.005-5000 mg/kg/day, and the dosage can be beyond the dosage range according to the severity of diseases or different dosage forms.
The compounds of the invention can be self-assembled into nanoparticles alone to improve activity, or in combination with other antineoplastic agents such as alkylating agents (e.g., cyclophosphamide or chlorambucil), antimetabolites (e.g., 5-fluorouracil or hydroxyurea), topoisomerase inhibitors (e.g., camptothecin), mitotic inhibitors (e.g., paclitaxel or vinblastine), DNA intercalators (e.g., doxorubicin) to improve activity, or in combination with radiation therapy. These other antineoplastic agents or radiation therapy may be administered simultaneously or at different times than the compounds of the present invention. These combination therapies may produce a synergistic effect that helps improve the therapeutic effect.
The invention combines the structural characteristics, structure-activity relationship and pharmacophore model of the anti-tumor drug 2-crotonyloxymethyl-2-cyclohexene (COMC-6) and the natural alkaloid beta-carboline with anti-tumor activity, adopts a prodrug strategy, selectively releases fluorescence in tumor cells through GSH/GST pi, degrades to generate active fragments, and realizes the enhancement of fluorescence signals and imaging detection. The research result of the inhibition effect of the compound on malignant tumor cells shows that the compound can carry out early diagnosis on cancer cells through fluorescent signals and can strongly inhibit the proliferation of various tumor cells such as liver cancer, colon cancer, cervical cancer and the like. The compound has the advantages of ingenious design, good GSH selectivity, high antitumor activity, low toxicity and the like. The fluorescent sensor is used as a sensor for intracellular GSH/GST pi fluorescence imaging, has the advantages of high selectivity, direct observation, easy real-time monitoring and the like, and can play an important role in the fields of detection, imaging and treatment of cancer cells.
Drawings
FIG. 1 shows a compound I1pH response uv fluorescence spectrum of (a). (FIG. 1A is Compound I1Ultraviolet absorption spectrum at pH 3-8; FIG. 1B is compound I1Fluorescence emission spectrum at pH 3-8 (Ex ═ 445 nm); FIG. 1C shows Compound I1The change curve of the ultraviolet absorbance at 445nm at pH 3-8; FIG. 1D is I1Change curve of fluorescence intensity at 490nm at pH 3-8).
FIG. 2 shows Compound I1GSH response uv fluorescence spectra of (a). FIG. 2A shows Compound I1Ultraviolet absorption spectrum of GSH (containing catalytic amount of GST pi) at about 0-10 equivalent; FIG. 2B is Compound I1Fluorescence emission spectrum of GSH (containing catalytic amount of GST pi) around 0-50 equivalents (Ex ═ 440 nm); FIG. 2C shows Compound I1Curve of fluorescence intensity at 492nm as a function of GSH concentration; FIG. 2D shows Compound I1Time dose curve of the fluorescence intensity of (c) with GSH (containing catalytic amounts of GST pi).
FIG. 3 shows Compound I1And the biological endogenous substance is specifically responded to the fluorescence characteristic.
FIG. 4 is a fluorescent image of the compounds of the present invention in HT29 cells.
FIG. 5 shows Compound I1In vivo fluorescence imaging.
FIG. 6 shows Compound I of the present invention1Results of antitumor activity in vivo.
Detailed Description
To further illustrate the present invention, a series of examples are given below, which are purely illustrative and are intended to be a detailed description of the invention only and should not be understood as limiting the invention.
Example 1 (6-Oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)1) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)1) Preparation of
Figure GDA0002639606720000101
(1) Preparation of 1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-carbohydrazide (2a)
After the compound methyl 1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-carboxylate (1a) (19.6g,50mmol) was dissolved in 80mL of methanol, 176.4mL of 85% hydrazine hydrate (75g,1.50mol) was added, reflux was carried out for 4-5H, TLC monitored reaction was complete, reaction solution was cooled to 0 ℃ and vacuum pump-filtered to obtain a light brown solid. A large amount of cold water is added into the filtrate, the solid is continuously precipitated, and after suction filtration and vacuum drying, 16.5g of light brown solid is obtained, and the yield is 84.2%.
(2) Preparation of 1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3a)
Compound (2a) (19.6g,50mmol) was dissolved in 80mL of 2mol/L HCl solution, followed by 60mL of H2O to NaNO2(10.4g,150mmol) was stirred under ice-bath conditions to prepare NaNO2The solution is slowly dropped into the solution of the compound (2a), and then after stirring for 1-4h, TLC monitors that the reaction is finished, and NaOH solution with the concentration of 1mol/L is used for adjusting the pH value to 7-8, a light yellow solid is precipitated, and the light yellow solid product 17.8g is obtained by suction filtration, and the yield is 88.4%.
(3) Preparation of 1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4a)
Dissolving compound (3a) (20.1g,50mmol) with 100mL of mixed solution of water and glacial acetic acid (1:1), refluxing at 90 deg.C for 4-5h, monitoring reaction by TLC, concentrating the reaction solution under reduced pressure to obtain sand, and purifying by column chromatography to obtain 8.4g light yellow solid with yield of48.2％。ESI-MS(m/z):350[M+H]+。
(4) Preparation of 4-nitrophenyl ((6-oxocyclohex-1-en-1-yl) methyl) carbonate (6b)
After completely dissolving the compound 2- (hydroxymethyl) -cyclohexenone (5b) (1.3g,10mmol) in 12ml of dichloromethane, N-Diisopropylethylamine (DIPEA) (3.9g,30mmol) was added, p-nitrophenyl chloroformate (3.1g,15mmol) was added under ice bath, stirring was carried out for 1-2h, the reaction was monitored by TLC to completion, the reaction solution was concentrated under reduced pressure to prepare sand, and column chromatography (EA: PE ═ 1:10) was carried out to obtain 1.8g of pale yellow liquid, with a yield of 62.1%.
(5) (6-Oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)1) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)1) Preparation of
Mixing compound 6(2.9g,10mmol) and 1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ]]Indol-3-amine (3.49g,10mmol) is dissolved in 20ml dichloromethane, stirred in ice bath, then DIPEA (3.9g,30mmol) is added, after 1-2h, the reaction is monitored by TLC until completion, the reaction solution is concentrated under reduced pressure to prepare sand, and the sand is purified by column chromatography (EA: PE ═ 1:4) to obtain a yellow solid (I)1)1.8g and (II)1)1.1g, yield 35.6% and 21.9%, respectively. (I)1) The spectrogram data is as follows: ESI-MS (M/z) 524[ M + Na ]]+；1H NMR(d6-DMSO,400MHz):10.83(s,1H,NH),8.07(d,J＝7.8Hz,1H,NH),7.46(q,J＝8.0Hz,2H,Ar-H),7.21(s,2H,Ar-H),7.13–7.09(m,2H,Ar-H),6.99(s,1H,Ar-H),6.15(d,J＝19.6Hz,1H,CH),4.10(s,2H,CH2),3.91(s,6H,CH3),3.76(s,3H,CH3),2.43–2.36(m,2H,CH2),2.36–2.30(m,2H,CH2),1.95–1.87(m,2H,CH2)。13C NMR(d6-DMSO,101MHz):199.32,153.40,152.53,145.79,143.03,138.17,137.03,128.37,127.91,126.62,121.98,121.31,112.48,106.11,95.58,60.54,56.32,38.55,25.67,23.18。(Ⅱ1) The spectrogram data is as follows: ESI-MS (M/z):654[ M + H]+；1H NMR(d6-DMSO,400MHz):10.89(s,1H,NH),8.15(d,J＝8.1Hz,1H,Ar-H),7.50–7.43(m,2H,Ar-H),7.23(s,2H,Ar-H),7.18(s,1H,Ar-H),7.13–7.09(m,1H,Ar-H),6.73(s,2H,CH),4.35(s,4H,CH2),3.89(s,6H,CH3),3.76(s,3H,CH3),2.42–2.37(m,4H,CH2),2.31(s,4H,CH2),1.92–1.90(m,4H,CH2)。13C NMR(d6-DMSO,101MHz):199.83,153.35,151.48,145.43,142.93,138.40,137.96,134.78,134.66,134.06,128.45,127.45,122.32,105.78,56.14,47.57,38.57,25.66,23.16。
Example 2 (6-Oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)2) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)2) Preparation of
Preparation of 1- (p-tolyl) -9H-pyrido [3,4-b ] indole-3-carbohydrazide (2b)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process with (1b), light yellow solid (2b) was finally obtained with a yield of 80.9%.
Preparation of 1- (p-tolyl) -9H-pyrido [3,4-b ] indole-3-ylazide (3b)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process with (2b), light yellow solid (3b) was obtained with 87.5% yield.
Preparation of 1- (p-tolyl) -9H-pyrido [3,4-b ] indol-3-amine (4b)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process with (3b), light yellow solid (4b) was obtained with 50.7% yield. ESI-MS (M/z):274[ M + H]+。
(6-Oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)2) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)2) Preparation of
Reference example 1 (I)1) The synthesis of (4a) is replaced by (4b), and a pale yellow solid (I) is obtained2) And (II)2) The yields were 34.1% and 22.3%, respectively.(I2) The spectrogram data is as follows: ESI-MS (M/z) 426[ M + H]+；1H NMR(d6-DMSO,400MHz):10.78(s,1H,NH),8.11(d,J＝7.8Hz,1H,NH),7.50–7.36(m,2H,Ar-H),7.28–7.20(m,3H,Ar-H),7.11–7.02(m,3H,Ar-H),6.94(s,1H,Ar-H),6.19(d,J＝19.5Hz,1H,CH),4.16(s,2H,CH2),3.03(s,3H,CH3),2.45–2.38(m,2H,CH2),2.36–2.32(m,2H,CH2),1.96–1.84(m,2H,CH2)。(Ⅱ2) The spectrogram data is as follows: ESI-MS (M/z):578[ M + H]+；1H NMR(d6-DMSO,400MHz):10.80(s,1H,NH),8.11(d,J＝8.1Hz,1H,Ar-H),7.59–7.47(m,2H,Ar-H),7.38–7.25(m,3H,Ar-H),7.21(s,1H,Ar-H),7.15–7.11(m,2H,Ar-H),6.79(s,2H,CH),4.39(s,4H,CH2),3.16(s,3H,CH3),2.41–2.35(m,4H,CH2),2.28(s,4H,CH2),1.95–1.92(m,4H,CH2)。
Example 3 (6-Oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)3) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)3) Preparation of
Preparation of 1- (3-methoxyphenyl) -9H-pyrido [3,4-b ] indole-3-carbohydrazide (2c)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process with (1c), light yellow solid (2c) was finally obtained with a yield of 81.3%.
Preparation of 1- (3-methoxyphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3c)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process with (2c), light yellow solid (3c) was obtained with 88.4% yield.
Preparation of 1- (3-methoxyphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4c)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process with (3c), a pale yellow solid (4c) was finally obtained with a yield of 52.3%. ESI-MS (M/z):290[ M + H]+。
(6-oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-Pyrido [3,4-b]Indol-3-yl carbamates (I)3) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)3) Preparation of
Reference example 1 (I)1) The synthesis of (4) is replaced by (4c) to obtain a pale yellow solid (I)3) And (II)3) The yields were 30.4% and 21.8%, respectively. (I)3) The spectrogram data is as follows: ESI-MS (M/z):442[ M + H]+；1H NMR(d6-DMSO,400MHz):10.79(s,1H,NH),8.04(s,1H,NH),7.43–7.39(m,3H,Ar-H),7.23–7.18(m,3H,Ar-H),7.15(s,2H,Ar-H),6.97(s,1H,Ar-H),6.13(d,J＝19.6Hz,1H,CH),4.08(s,2H,CH2),3.88(s,3H,CH3),2.41–2.35(m,2H,CH2),2.33–2.29(m,2H,CH2),1.94–1.85(m,2H,CH2)。(Ⅱ3) The spectrogram data is as follows: ESI-MS (M/z):594[ M + H]+；1H NMR(d6-DMSO,400MHz):10.87(s,1H,NH),8.14(d,J＝8.0Hz,1H,Ar-H),7.48–7.42(m,3H,Ar-H),7.25(s,2H,Ar-H),7.19(s,1H,Ar-H),7.11–7.08(m,2H,Ar-H),6.71(s,2H,CH),4.33(s,4H,CH2),3.85(s,6H,CH3),2.41–2.35(m,4H,CH2),2.31–2.19(m,4H,CH2),1.94–1.89(m,4H,CH2)。
Example 4 (6-Oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)4) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)4) Preparation of 1- (4-methoxyphenyl) -9H-pyrido [3,4-b ]]Preparation of indole-3-carbonyl hydrazide (2d)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process with (1d), light yellow solid (2d) was finally obtained with a yield of 82.0%.
Preparation of 1- (4-methoxyphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3d)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process with (2d), a pale yellow solid (3d) was finally obtained in 87.4% yield.
Preparation of 1- (4-methoxyphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4d)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process with (3d), a pale yellow solid (4d) was finally obtained with a yield of 52.5%. ESI-MS (M/z):290[ M + H]+。
(6-Oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)4) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)4) Preparation of
Reference example 1 (I)1) The synthesis of (4) is replaced by (4d) to obtain a pale yellow solid (I)4) And (II)4) The yields were 35.1% and 22.6%, respectively. (I)4) The spectrogram data is as follows: ESI-MS (M/z):442[ M + H]+；1H NMR(d6-DMSO,400MHz):10.83(s,1H,NH),8.04(d,J＝7.8Hz,1H,NH),7.44–7.39(m,2H,Ar-H),7.24–7.17(m,3H,Ar-H),7.14–7.08(m,2H,Ar-H),6.97(s,2H,Ar-H),6.12(d,J＝19.6Hz,1H,CH),4.12(s,2H,CH2),3.83(s,3H,CH3),2.41–2.34(m,2H,CH2),2.35–2.32(m,2H,CH2),1.98–1.87(m,2H,CH2)。(Ⅱ4) The spectrogram data is as follows: ESI-MS (M/z):594[ M + H]+；1H NMR(d6-DMSO,400MHz):10.90(s,1H,NH),8.17(s,1H,Ar-H),7.55–7.48(m,2H,Ar-H),7.33–7.29(m,2H,Ar-H),7.22(s,2H,Ar-H),7.18–7.10(m,2H,Ar-H),6.77(s,2H,CH),4.37(s,4H,CH2),3.80(s,6H,CH3),2.41–2.37(m,4H,CH2),2.30–2.19(m,4H,CH2),1.94–1.90(m,4H,CH2)。
Example 5 (6-Oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)5) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)5) Preparation of
Preparation of 1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indole-3-carbonyl hydrazide (2e)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process with (1e), red solid (2e) was finally obtained with a yield of 81.1%.
Preparation of 1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3e)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process with (2e), a brown solid (3e) was finally obtained in 80.9% yield.
Preparation of 1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4e)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process with (3e), red solid (4e) was finally obtained with a yield of 45.8%. ESI-MS (M/z):303[ M + H]+。
(6-Oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)5) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)5) Preparation of
Reference example 1 (I)1) By replacing (4a) in the process with (4e), to give a pale red solid (I)5) And (II)5) The yields were 32.7% and 19.1%, respectively. (I)5) The spectrogram data is as follows: ESI-MS (M/z):455[ M + H]+；1H NMR(d6-DMSO,400MHz):10.88(s,1H,NH),8.09(s,1H,NH),7.51–7.44(m,3H,Ar-H),7.26–7.18(m,3H,Ar-H),7.12(s,2H,Ar-H),6.99(s,1H,Ar-H),6.19(d,J＝19.6Hz,1H,CH),4.14(s,2H,CH2),3.99(s,6H,CH3),2.45–2.37(m,2H,CH2),2.38–2.32(m,2H,CH2),1.97–1.89(m,2H,CH2)。(Ⅱ5) The spectrogram data is as follows: ESI-MS (M/z):607[ M + H]+；1H NMR(d6-DMSO,400MHz):10.95(s,1H,NH),8.18(s,1H,Ar-H),7.50–7.43(m,3H,Ar-H),7.30–7.24(m,2H,Ar-H),7.19(s,2H,Ar-H),7.14–7.10(m,1H,Ar-H),6.79(s,2H,CH),4.41(s,4H,CH2),4.04(s,6H,CH3),2.46(s,4H,CH2),2.31–2.25(m,4H,CH2),1.94–1.85(m,4H,CH2)。
EXAMPLE 6 (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)6) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)6) Preparation of
Preparation of 1- (2, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-carbohydrazide (2f)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process with (1f), a pale yellow solid (2f) was finally obtained in 87.1% yield.
Preparation of 1- (2, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3f)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process with (2f), a pale yellow solid (3f) was finally obtained with a yield of 86.2%.
Preparation of 1- (2, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4f)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process with (3f), a pale yellow solid (4f) was finally obtained in 55.2% yield. ESI-MS (M/z) 320[ M + H]+。
(6-Oxocyclohex-1-en-1-yl) methyl (1- (2, 4-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)6) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)6) Preparation of
Reference example 1 (I)1) The synthesis of (4) is replaced by (4f) to obtain a pale yellow solid (I)6) And (II)6) The yields were 34.2% and 20.7%, respectively. (I)6) The spectrogram data is as follows: ESI-MS (M/z):472[ M + H]+；1H NMR(d6-DMSO,400MHz):10.79(s,1H,NH),8.10(s,1H,NH),7.47–7.36(m,3H,Ar-H),7.28(s,1H,Ar-H),7.19–7.10(m,3H,Ar-H),7.02(s,1H,Ar-H),6.18(d,J＝19.6Hz,1H,CH),4.19(s,2H,CH2),3.93(s,3H,CH3),3.78(s,3H,CH3),2.37–2.30(m,2H,CH2),2.27–2.21(m,2H,CH2),1.93–1.82(m,2H,CH2)。(Ⅱ6) The spectrogram data is as follows: ESI-MS (M/z):624[ M + H]+；1H NMR(d6-DMSO,400MHz):10.95(s,1H,NH),8.05(s,1H,Ar-H),7.55–7.46(m,3H,Ar-H),7.26(s,1H,Ar-H),7.19(s,1H,Ar-H),7.16–7.11(m,2H,Ar-H),6.78(s,2H,CH),4.39(s,4H,CH2),3.93(s,3H,CH3),3.80(s,3H,CH3),2.45–2.36(m,4H,CH2),2.23–2.15(m,4H,CH2),1.90–1.82(m,4H,CH2)。
EXAMPLE 7 (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)7) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)7) Preparation of
Preparation of 1- (2, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-carbohydrazide (2g)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process with (1g), a pale yellow solid (2g) was finally obtained with a yield of 85.1%.
Preparation of 1- (2, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3g)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process with (2g), a pale yellow solid (3g) was obtained in 86.9% yield.
Preparation of 1- (2, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4g)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process with (3g), a pale yellow solid (4g) was finally obtained in 52.4% yield. ESI-MS (M/z) 320[ M + H]+。
(6-Oxocyclohex-1-en-1-yl) methyl (1- (2, 5-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)7) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)7) Preparation of
Reference example 1 (I)1) The synthesis of (4) from (4g instead of (4a) in the process, gives a pale yellow colorSolid (I)7) And (II)7) The yields were 35.7% and 18.4%, respectively. (I)7) The spectrogram data is as follows: ESI-MS (M/z):472[ M + H]+；1H NMR(d6-DMSO,400MHz):10.82(s,1H,NH),8.04(s,1H,NH),7.45–7.34(m,3H,Ar-H),7.23(s,2H,Ar-H),7.14–7.02(m,2H,Ar-H),6.98(s,1H,Ar-H),6.27(d,J＝19.6Hz,1H,CH),4.21(s,2H,CH2),3.92(s,3H,CH3),3.79(s,3H,CH3),2.43–2.35(m,2H,CH2),2.33–2.21(m,2H,CH2),1.96–1.86(m,2H,CH2)。(Ⅱ7) The spectrogram data is as follows: ESI-MS (M/z):624[ M + H]+；1H NMR(d6-DMSO,400MHz):10.89(s,1H,NH),8.12(s,1H,Ar-H),7.62–7.51(m,2H,Ar-H),7.31–7.22(m,3H,Ar-H),7.11–7.05(m,2H,Ar-H),6.81(s,2H,CH),4.29(s,4H,CH2),3.92(s,3H,CH3),3.66(s,3H,CH3),2.52–2.46(m,4H,CH2),2.30–2.18(m,4H,CH2),1.93–1.89(m,4H,CH2)。
EXAMPLE 8 (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)8) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)8) Preparation of
Preparation of 1- (3, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-carbohydrazide (2H)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process by (1h), a pale yellow solid (2h) was finally obtained with a yield of 84.5%.
Preparation of 1- (3, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3H)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process with (2h), a pale yellow solid (3h) was obtained in 86.7% yield.
Preparation of 1- (3, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4H)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process by (3h), a pale yellow solid (4h) was obtained in 51.7% yield. ESI-MS (m-z):320[M+H]+。
(6-Oxocyclohex-1-en-1-yl) methyl (1- (3, 4-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)8) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 4-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)8) Preparation of
Reference example 1 (I)1) (4h) instead of (4a) in the process, to give a pale yellow solid (I)8) And (II)8) The yields were 36.1% and 16.7%, respectively. (I)8) The spectrogram data is as follows: ESI-MS (M/z):472[ M + H]+；1H NMR(d6-DMSO,400MHz):10.77(s,1H,NH),8.01(d,J＝7.8Hz,1H,NH),7.50–7.39(m,3H,Ar-H),7.27(s,2H,Ar-H),7.15–7.09(m,2H,Ar-H),6.98(s,1H,Ar-H),6.16(d,J＝19.6Hz,1H,CH),4.22(s,2H,CH2),3.93(s,3H,CH3),3.78(s,3H,CH3),2.42–2.35(m,2H,CH2),2.31–2.23(m,2H,CH2),1.93–1.87(m,2H,CH2)。(Ⅱ8) The spectrogram data is as follows: ESI-MS (M/z):624[ M + H]+；1H NMR(d6-DMSO,400MHz):10.98(s,1H,NH),8.09(s,1H,Ar-H),7.55–7.46(m,3H,Ar-H),7.28(s,2H,Ar-H),7.19(s,1H,Ar-H),7.15–7.08(m,1H,Ar-H),6.81(s,2H,CH),4.33(s,4H,CH2),3.84(s,3H,CH3),3.73(s,3H,CH3),2.41–2.36(m,4H,CH2),2.29–2.24(s,4H,CH2),1.92–1.88(m,4H,CH2)。
Example 9 (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)9) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)9) Preparation of
Preparation of 1- (3, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-carbohydrazide (2i)
Referring to the synthesis of (2a) in example 1, replacing (1a) in the process by (1i), light yellow solid (2i) was finally obtained with a yield of 81.0%.
Preparation of 1- (3, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indole-3-acyl azide (3i)
Referring to the synthesis of (3a) in example 1, replacing (2a) in the process by (2i), a pale yellow solid (3i) was finally obtained with a yield of 85.7%.
Preparation of 1- (3, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-amine (4i)
Referring to the synthesis of (4a) in example 1, replacing (3a) in the process by (3i), a pale yellow solid (4i) was finally obtained with a yield of 53.2%. ESI-MS (M/z) 320[ M + H]+。
(6-Oxocyclohex-1-en-1-yl) methyl (1- (3, 5-Dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)9) Or bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 5-dimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)9) Preparation of
Reference example 1 (I)1) (4i) instead of (4a) in the process, to give (I) as a pale yellow solid9) And (II)9) The yields were 34.4% and 24.1%, respectively. (I)9) The spectrogram data is as follows: ESI-MS (M/z):472[ M + H]+；1H NMR(d6-DMSO,400MHz):10.82(s,1H,NH),8.06(d,J＝7.8Hz,1H,NH),7.49–7.39(m,3H,Ar-H),7.24(s,2H,Ar-H),7.13–7.08(m,2H,Ar-H),6.98(s,1H,Ar-H),6.52(d,J＝19.6Hz,1H,CH),4.09(s,2H,CH2),3.92(s,6H,CH3),2.43–2.35(m,2H,CH2),2.36–2.31(m,2H,CH2),1.96–1.87(m,2H,CH2)。(Ⅱ9) The spectrogram data is as follows: ESI-MS (M/z):624[ M + H]+；1H NMR(d6-DMSO,400MHz):10.85(s,1H,NH),8.12(d,J＝8.0Hz,1H,Ar-H),7.52–7.46(m,3H,Ar-H),7.22(s,2H,Ar-H),7.19(s,1H,Ar-H),7.14–7.08(m,1H,Ar-H),6.76(s,2H,CH),4.38(s,4H,CH2),3.87(s,6H,CH3),2.42–2.36(m,4H,CH2),2.27–2.19(s,4H,CH2),1.91–1.86(m,4H,CH2)。
Example 10 (6-Oxocyclopent-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)10) Or bis (6-oxocyclopent)-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)10)；
Preparation of 4-nitrophenyl ((6-oxocyclopent-1-en-1-yl) methyl) carbonate (6a)
Referring to the synthesis method of (6b) in example 1, replacing (5b) in the method with (5a), light yellow liquid (6a) was finally obtained with a yield of 85.1%.
(6-Oxocyclopent-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)10) Or bis (6-oxocyclopent-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)10) Preparation of
Reference example 1 (I)1) (6a) in place of (6b) in the process to give a pale yellow solid (I)10) And (II)10) The yields were 30.7% and 19.6%, respectively. (I)10) The spectrogram data is as follows: ESI-MS (M/z):488[ M + H [)]+；1H NMR(d6-DMSO,400MHz):10.83(s,1H,NH),8.08(d,J＝7.8Hz,1H,NH),7.45–7.36(m,2H,Ar-H),7.24(s,2H,Ar-H),7.16–7.09(m,2H,Ar-H),6.99(s,1H,Ar-H),6.18(d,J＝19.6Hz,1H,CH),4.14(s,2H,CH2),3.92(s,6H,CH3),3.78(s,3H,CH3),2.35–2.29(m,2H,CH2),1.96–1.87(m,2H,CH2)。(Ⅱ10)ESI-MS(m/z):626[M+H]+。
Example 11 (6-Oxocyclopent-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)11) Or bis (6-oxocyclopent-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)11) Preparation of
Reference example 1 (I)1) The synthesis of (4) and (6b) in the (4e) and (6a) replacement processes gives a pale red solid (I)11) And (II)11) The yields were 31.2% and 18.4%, respectively. (I)11) The spectrogram data is as follows: ESI-MS (M/z):441[ M + H]+；1H NMR(d6-DMSO,400MHz):10.87(s,1H,NH),8.04(d,J＝7.8Hz,1H,NH),7.44–7.37(m,3H,Ar-H),7.24(s,2H,Ar-H),7.16–7.11(m,3H,Ar-H),6.98(s,1H,Ar-H),6.17(d,J＝19.6Hz,1H,CH),4.19(s,2H,CH2),4.11(s,6H,CH3),2.38–2.32(m,2H,CH2),1.97–1.88(m,2H,CH2)。(Ⅱ11) The spectrogram data is as follows: ESI-MS (M/z):579[ M + H]+。
Example 12 (6-oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)12) Or bis (6-oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)12)；
Preparation of 4-nitrophenyl ((6-oxocyclopent-1-en-1-yl) methyl) carbonate (6c)
Referring to the synthesis method of (6c) in example 1, replacing (5b) in the method with (5c), light yellow liquid (6c) was finally obtained in 84.0% yield.
(6-Oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)12) Or bis (6-oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)12) Preparation of
Reference example 1 (I)1) (6c) in place of (6b) in the process, to give (I) as a yellow solid12) And (II)12) The yields were 29.8% and 18.0%, respectively. (I)12) The spectrogram data is as follows: ESI-MS (M/z):516[ M + H]+；1H NMR(CDCl3,400MHz):8.11(s,1H,NH),8.03(d,J＝7.8Hz,1H,Ar-H),7.50–7.46(m,1H,Ar-H),7.40(d,J＝8.1Hz,1H,Ar-H),7.22–7.19(m,1H,Ar-H),7.14(s,2H,Ar-H),6.93(s,1H,Ar-H),6.87–6.84(m,1H,CH),4.21(s,2H,CH2),3.95(s,6H,CH3),3.92(s,3H,CH3),2.70–2.58(m,2H,CH2),2.44–2.40(m,2H,CH2),1.82–1.71(m,4H,CH2)。13C NMR(CDCl3,101MHz):205.05,153.77,152.78,143.12,141.80,140.13,138.52,134.37,133.69,128.47,128.22,122.01,121.90,119.45,111.45,105.48,94.46,60.99,56.44,45.35,42.90,27.65,25.18,21.49。(Ⅱ12)ESI-MS(m/z):682[M+H]+。
Example 13 (6-oxocyclohept-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (I)13) Or bis (6-oxocyclohept-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3, 4-b)]Indol-3-yl carbamates (II)13) Preparation of
Reference example 1 (I)1) The synthesis of (4) and (6b) in the processes are replaced by (4e) and (6c), and finally a light red solid (I) is obtained13) And (II)13) The yields were 27.7% and 16.9%, respectively. (I)13)；1H NMR(d6-DMSO,400MHz):10.94(s,1H,NH),8.08(d,J＝7.8Hz,1H,NH),7.47–7.39(m,3H,Ar-H),7.23(s,2H,Ar-H),7.15–7.10(m,3H,Ar-H),7.01(s,1H,Ar-H),6.18(d,J＝19.6Hz,1H,CH),4.23(s,2H,CH2),4.16(s,6H,CH3),2.73–2.66(m,2H,CH2),2.46–2.40(m,2H,CH2),1.89–1.82(m,4H,CH2)。ESI-MS(m/z):469[M+H]+。(Ⅱ13)ESI-MS(m/z):635[M+H]+。
EXAMPLE 14 examination of tumor cell proliferation inhibition assay of the Compound of the present invention by MTT method
The anti-proliferation activity of the compound on 4 human cancer cell lines is evaluated by adopting a tetramethyl triazole blue colorimetric Method (MTT) in-vitro anti-tumor test. COMC-6 was used as a positive control. Human cancer cell line: liver cancer cell HepG2, human cervical carcinoma cell Hela, colon cancer cell HCT116 and HT29 cell, and stomach cancer cell HGC-27.
The experimental method is as follows: taking a bottle of cells in exponential growth phase, adding 0.25% trypsin for digestion to make adherent cells fall off, and preparing the solution containing 2 × 10 cells per ml4～4×104A suspension of individual cells. Inoculating the cell suspension on a 96-well plate, placing 180 μ L of the cell suspension in each well, and placing in a constant temperature CO2The culture was carried out in an incubator for 24 hours. Changing the solution, adding the test compound I1-Ⅰ13And II1-Ⅱ13(Compounds dissolved in DMSO and diluted with PBSThe concentrations of the test compounds were 6.25X 10, respectively-6，1.25×10-5，2.5×10-5，5×10-5mol/L), 20. mu.L per well, and cultured for 72 hours. MTT was added to a 96-well plate at 20. mu.L per well and reacted in an incubator for 4 hours. The supernatant was aspirated, DMSO was added, 150. mu.L per well, and shaken on a plate shaker for 5 minutes. The absorbance of each well was measured at a wavelength of 570nm using an enzyme linked immunosorbent assay to calculate the cell inhibition rate. The results of the experiment are shown in table 2.
The cell inhibition rate (negative control OD value-test substance OD value)/negative control OD value × 100%.
The compounds of the invention are tested by a series of tumor cell antiproliferative activity, and pharmacological experiment results (shown in table 2) show that the I or II series compounds of the invention have stronger inhibition effect on the proliferation of most tumor cells, and especially, part of the compounds are slightly stronger or equivalent to the positive control drug COMC-6.
TABLE 2 inhibition of human tumor cells by partial compounds of the invention (% 12.5. mu. mol/L)
Figure GDA0002639606720000211
Figure GDA0002639606720000221
ND: not detected
EXAMPLE 15 pH-sensitive fluorescence characteristics of Compounds of the invention
The ultraviolet absorption wavelength and the change condition of fluorescence thereof along with pH of the I or II series compounds of the invention are measured by an ultraviolet-visible spectrophotometer and a fluorescence spectrometer. The selected pH range is 3.1-7.6.
The pH sensitive uv absorption test method is as follows: 100 μ M of the series of compounds was dissolved in 1% aqueous DMSO solution at a pH of about 8.00-3.00. All absorption spectra were recorded at room temperature with a scanning wavelength range of 350-600nm and a scanning speed of 1.0 nm/s.
The pH sensitive fluorescence experiment method is as follows: mu.M of the series of compounds was dissolved in 1% DMSO aqueous solution at a pH of about 8.00-3.00. All emission spectra were performed at room temperature, with 430-470nm excitation, recorded at 450nm to 650 nm.
The results show that the compounds I of the invention1-Ⅰ13And II1-Ⅱ13The characteristic ultraviolet absorption wavelength of the ultraviolet absorption material is 390-420 nm, the peak value of the ultraviolet absorption material is gradually reduced along with the reduction of the pH value, and the peak value at the position of 430-470nm is gradually increased; the fluorescence spectrum shows that the fluorescence at 480-520 nm also increases obviously along with the reduction of the pH value.
Wherein the compound I1The typical pH response ultraviolet fluorescence spectrum is shown in FIG. 1, the characteristic ultraviolet absorption wavelength is around 402nm, and the peak value at 402nm gradually decreases (FIG. 1A) and the peak value at 445nm gradually increases (FIG. 1A and FIG. 1C) along with the decrease of the pH value; its fluorescence at 490nm also gradually increased with decreasing pH value ((fig. 1B and fig. 1D, excitation wavelength Ex 445 nm).
Example 16 Compounds of the invention and GSH-responsive fluorescence characteristics
The changes of ultraviolet and fluorescence of the I-II series compounds of the invention along with the concentration of GSH are judged by an extracellular fluorescence experiment.
The GSH combined ultraviolet absorption experimental method is as follows: 100 μ M of the series was dissolved in 1% aqueous DMSO, to which approximately 0-10 equivalents of GSH and catalytic amounts of GST π were added. All absorption spectra were recorded after incubation at 37 ℃ with a scanning wavelength range of 350-600nm and a scanning speed of 1.0 nm/s.
GSH binding fluorescence assay methods were as follows: mu.M of the series was dissolved in 1% aqueous DMSO, to which approximately 0-50. mu.M GSH and catalytic amounts of GST π were added. All emission spectra were recorded after incubation at 37 ℃ for 0.5 h, 430-470nm excitation, 450nm to 650 nm.
Compounds of the invention I1-Ⅰ13And II1-Ⅱ13Under the concentration of 1 mu M, with the increase of GSH equivalent, the peak value of the characteristic ultraviolet absorption wavelength is gradually reduced between 385 and 430nm, and the peak value at 435 to 470nm is gradually increased;the fluorescence spectrum shows that the fluorescence at 470-530 nm also gradually increases with the increase of GSH equivalent.
Wherein the compound I1The response ultraviolet fluorescence spectrum of the representative GSH is shown in figure 2, the ultraviolet absorption wavelength is around 402nm, and the peak value at 402nm gradually decreases and the peak value at 440nm gradually increases with the increase of the concentration of the GSH (figure 2A); fluorescence was released immediately after GSH addition and peaked rapidly (fig. 2D, excitation wavelength Ex ═ 440nm), with an emission wavelength of 492nm, and fluorescence intensity increased with increasing GSH concentration (fig. 2B and 2C).
Example 17 specific response of Compounds of the invention to GSH fluorescence characteristics
Specific binding of the compounds of the invention to GSH was judged by extracellular fluorescence experiments.
The specific experimental method is as follows: mu.M of the series of compounds are dissolved in an aqueous solution containing a catalytic amount of GST pi, to which 1mM of K is added+、Na+、Ca2+、Mg2+、Zn2+、Al3+、Cu2+、Fe2+(ii) a 100 μ M GSH, lysine, histidine, alanine, cysteine, glutamic acid, serine, glycine, arginine, vitamin C, Na2S、H2O2And NADH, and the like. All emission spectra were recorded after incubation at 37 ℃ for 0.5 h, 430-480nm excitation, 450-650 nm.
With a compound I1As representative, compound I1The chemoselectivity under GSH is far superior to other common components or ions in biological systems, such as K+、Na+、Ca2+、Mg2+、Zn2+、Al3+、Cu2+、Fe2+Lysine, histidine, alanine, cysteine, glutamic acid, serine, glycine, arginine, vitamin C, Na2S、H2O2NADH, and the like. The results are shown in FIG. 3, where I is the presence of other analytes1It is very stable, and its fluorescence intensity at 492nm is still very weak and unchanged.
Example 18 tumor cell imaging Using confocal microscopy
A fluorescence imaging method of tumor cells by using a confocal microscope selects the HT29 cells with high expression of GST pi, and comprises the following steps:
(1) centrifuging HT29 cell suspension at 3000 rpm for 5min, and removing supernatant;
(2) adding the prepared 1uM PBS solution containing the compound, incubating at the constant temperature of 37 ℃, and then carrying out a laser confocal scanning imaging experiment;
(3) placing the incubated cells on an objective table of a confocal microscope, and exciting the wavelength: 430-480 nm; receiving wavelength: 480-550 nm.
Confocal fluorescence images showing HT29 cells vs Compound I of the invention1-Ⅰ13And II1-Ⅱ13The absorption is rapid, relatively increases in the first 10min, gradually increases, the increase is brightest after 1h, and lasts for 2h, and the accumulation efficiency is high. FIG. 4 shows the representation of compound I in HT29 cells1、Ⅰ3、Ⅰ6、Ⅰ9、Ⅰ10、Ⅰ13、Ⅱ2、Ⅱ4、Ⅱ7、Ⅱ9、Ⅱ11、Ⅱ12The results of the cell fluorescence imaging pictures in 1h show that the compound can well perform fluorescence imaging in tumor cells.
EXAMPLE 19 in vivo imaging Studies of Compounds of the invention
To evaluate the in vivo imaging efficacy of the I-II series of compounds, BALB/c nude mice model was constructed by subcutaneous inoculation of HT29 cells, and the inventive compounds I were studied using a Living body imager1、Ⅰ3、Ⅰ6、Ⅰ9、Ⅰ10、Ⅰ13、Ⅱ2、Ⅱ4、Ⅱ7、Ⅱ9、Ⅱ11、Ⅱ12Fluorescence distribution of tissues in tumor-bearing nude mice. The compound of the present invention was administered intravenously at 40mg/kg, and 2 hours later, the mice were anesthetized, and the fluorescence signals of the tumor and major organs in the nude mice were examined.
The results show that the compounds of the inventionⅠ1、Ⅰ3、Ⅰ6、Ⅰ9、Ⅰ10、Ⅰ13、Ⅱ2、Ⅱ4、Ⅱ7、Ⅱ9、Ⅱ11、Ⅱ12The fluorescence intensity of tumor tissues in nude mice was significantly higher than that of major organs including heart, lung, liver, kidney and spleen. FIG. 5 shows Compound I of the present invention1The in vivo fluorescence imaging distribution result shows that the compound of the invention can selectively display stronger fluorescence signals in vivo tumor tissues.
Example 20 in vivo antitumor Activity Studies
To evaluate the in vivo antitumor activity of the series I-II compounds, BALB/c nude mouse model was established subcutaneously inoculated with HT29 cells. After solid tumors had formed, nude mice were randomly given PBS, control drug COMC-6, Compound I1Changes in tumor size were monitored every three days over 21 days.
Compounds of the invention I1The results of the in vivo antitumor activity study are shown in fig. 6, and stable tumor growth was observed in the control group. However, the compounds I1The treatment group significantly reduced the volume and weight of colon tumors. With the same dosage of I1The treatment showed better antitumor activity than COMC-6 treatment, in I1Greater tumor suppression rates resulted at the end of treatment. I compared with PBS-treated control group (1.49. + -. 0.27g)1The tumor weight (0.48. + -. 0.07g) of 40mg/kg treated mice was reduced by 67.7% (w/w), while the tumor weight (0.81. + -. 0.12g) of COMC-640 mg/kg treated control group was reduced by 45.6% (w/w). The results show that the compound I1Has significant anti-tumor activity on colon tumor growth in vivo.

Claims (9)

1. The beta-carboline-cycloenone derivative has a structure shown in the following general formula:
Figure FDA0002697186800000011
wherein,
R1represents a correspondingly substituted ringOne or more substituent(s) selected from H, C1-C6 alkoxy, C1-C6 alkyl, and C1-C6 alkylamino, R1When a plurality of substituents are represented, each substituent may be the same or different;
R2represents one or more substituents on the corresponding substituted ring, selected from H or C1-C6 alkyl, R2When a plurality of substituents are represented, each substituent may be the same or different;
R3selected from H or C1-C6 alkyl;
R4selected from alkyl groups of H, C1-C6 or
Figure FDA0002697186800000012
n is 1, 2 or 3.
2. The beta-carboline-cycloalkenone derivative according to claim 1, characterized in that said R1One or more selected from H, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, methylamino, ethylamino, methylethylamino and N, N-dimethylamino;
R2one or more selected from H, methyl, ethyl, propyl and isopropyl;
R3selected from H, methyl, ethyl, propyl or isopropyl;
R4selected from H, methyl, ethyl, propyl, isopropyl or
Figure FDA0002697186800000013
n is 1, 2 or 3.
3. The beta-carboline-cycloalkenone derivative according to claim 1, characterized in that said R1Represents 3,4, 5-trimethoxy, 4-CH3、3-OCH3、4-OCH34-N, N-dimethyl, 2, 4-dimethoxy, 2, 5-dimethoxy, 3, 4-dimethoxy or 3, 5-dimethoxy; r2Or R3Represents H; r4Represents H or
Figure FDA0002697186800000021
n is 1, 2 or 3.
4. The β -carboline-cycloalkenone derivative according to claim 1, characterized by being selected from the following compounds:
(6-oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (p-tolyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3-methoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-methoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (2, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (2, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (2, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (3, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 4-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohex-1-en-1-yl) methyl (1- (3, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohex-1-en-1-yl) methyl (1- (3, 5-dimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclopent-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclopent-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclopent-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclopent-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohept-1-en-1-yl) methyl (1- (3,4, 5-trimethoxyphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
(6-oxocyclohept-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate;
bis (6-oxocyclohept-1-en-1-yl) methyl (1- (4-N, N-dimethylphenyl) -9H-pyrido [3,4-b ] indol-3-yl) carbamate.
5. The process for producing the β -carboline-cycloalkenone derivative according to any one of claims 1 to 4, characterized by comprising the steps of:
(1) reacting the compound 1 in hydrazine hydrate to obtain a compound 2,
Figure FDA0002697186800000041
(2) reacting compound 2 with NaNO2Reacting under an acidic condition to obtain a compound 3,
Figure FDA0002697186800000042
(3) reacting the compound 3 under an acidic condition to obtain a compound 4;
Figure FDA0002697186800000043
(4) reacting the compound 5 with p-nitrophenyl chloroformate under alkaline condition to obtain a compound 6,
Figure FDA0002697186800000044
(5) reacting the compound 4 with the compound 6 under alkaline conditions to obtain a compound 7 and/or a compound 8,
Figure FDA0002697186800000045
or the synthesis step also comprises the step (6), the compound 7 is reacted under sodium hydrogen, and then is reacted with C1-C6 alkyl bromide to obtain the compound 9,
Figure FDA0002697186800000051
R1represents one or more substituents on corresponding substituted ring, and is selected from one or more of H, C1-C6 alkoxy, C1-C6 alkyl and C1-C6 alkylamino, R1When a plurality of substituents are represented, each substituent may be the same or different;
R2represents one or more substituents on the corresponding substituted ring, selected from H or C1-C6 alkyl, R2When a plurality of substituents are represented, each substituent may be the same or different;
R3selected from H or C1-C6 alkyl;
R4alkyl selected from C1-C6;
n is 1, 2 or 3.
6. Use of a β -carboline-cycloalkenone derivative according to any one of claims 1 to 4 for producing a drug and/or a probe targeting GST pi.
7. The use according to claim 6, characterized in that the GST pi-targeting drug is a drug for the treatment and/or prevention of cancer.
8. Use according to claim 7, characterized in that said cancer is selected from liver cancer, colon cancer, cervical cancer or stomach cancer.
9. The use according to claim 6, characterized in that the probe targeting GST pi is a fluorescent probe based on a dual pH and GSH response.
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