Indole heterocyclic compound, pharmaceutical composition and synthetic method and application thereof

Abstract

The invention provides an indole heterocyclic compound as shown in a formula (I) and / or a formula (II) or pharmaceutically acceptable salt, a pharmaceutical composition, a synthetic method and application thereof. The compound has good anti-tumor activity, the synthesis method of the compound is simple and convenient, and the compound shown in the formula (I) and / or the formula (II) can be selectively synthesized by selecting different catalyst types and regulating the proportion of substrates. The compound can be further used for preparing a reduced, acylated or ketoximated derivative compound thereof.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of chemical synthesis, in particular, to indole heterocyclic compounds, pharmaceutical compositions and synthetic methods and applications thereof. BACKGROUND

[0002] Nitrogen-containing or oxygen-containing heterocyclic compounds generally have excellent biological activity and drug activity, and they are the basic skeleton or mother nucleus for constructing pharmacophores. By improving the structure of these heterocyclic compounds, high-efficiency and selective drug molecules can be obtained. Indole and its derivatives belong to an important class of nitrogen-containing heterocyclic compounds, which are widely present in natural bioactive molecules, drugs and functional materials. Therefore, a lot of efforts have been devoted to developing methods for preparing indole skeletons. Indole derivatives with indole as the mother nucleus include bis-indole derivatives, heterocyclic indole derivatives, oxidized indole derivatives, hydrazine indole derivatives, amide indole derivatives, etc. Indole and its derivatives have anticancer, antituberculosis, antibacterial, antiviral, antimalarial, anti-inflammatory activity, anti-leishmaniasis drugs, anticholinesterase and enzyme inhibition effects. Connecting indole with heterocyclic structures with pharmacological effects to improve biological activity is a strategy widely used in the design of drug structures, among which the 3-position of indole is the most modified site. According to research reports, indole is connected with pyrrole, oxazole, thiazole, imidazole or quinoline heterocycle at 2, 3 or 5 positions to construct a series of indole-heterocyclic skeleton structures, and on this basis, new drug molecules are derived, which have obvious inhibitory effect on SMMC-7721, HL-60, HCT-29 and other cancer cells. At the same time, it is also widely used in the treatment of antibacterial, antiviral and antihypertensive fields. Therefore, as the most common design method of indole drugs, constructing indole-heterocyclic skeleton structure has very broad application prospects in the fields of organic synthesis, biochemistry and medicinal chemistry. SUMMARY

[0003] To improve the above technical problems, the present application provides a compound represented by formula (I) and / or formula (II) or a pharmaceutically acceptable salt thereof:

[0004]

[0005] wherein R is selected from the following groups which are unsubstituted or optionally substituted by one, two or more R 2 C1-6alkyl, C 1-10 C1-6alkoxy, C 1-10 C1-6alkyl, C 3-10 C1-6alkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl;

[0006] each R 1 is the same or different, and is independently selected from H, halogen, OH, CN, NO2, unsubstituted or optionally substituted by one, two or more Ra substituted lower alkyl, C 1-10 alkyl, C 1-10 alkoxy, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl;

[0007] each R 2 are the same or different, independently of each other, selected from the group consisting of H, halogen, OH, CN, NO2, unsubstituted or optionally substituted lower alkyl, C b substituted lower alkyl, C 1-10 alkyl, C 1-10 alkoxy, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl;

[0008] each R 3 are the same or different, independently of each other, selected from the group consisting of H, halogen, CN, NO2, unsubstituted or optionally substituted lower alkyl, C c substituted lower alkyl, C 1-10 alkyl, C 1-10 alkoxy, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl, NH2;

[0009] each R 4 are the same or different, independently of each other, selected from the group consisting of H, unsubstituted or optionally substituted lower alkyl, C d substituted lower alkyl, C 1-10 alkyl, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl;

[0010] each R a , R b , R c , R d are the same or different, independently of each other, selected from the group consisting of H, halogen, OH, CN, NO2, C 1-10 alkyl, C 1-10 alkoxy, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl, C 1-6 alkyl-C 6-10 aryl-S(O)2-;

[0011] m is selected from 0, 1, 2, 3, 4 or 5;

[0012] n is selected from 0, 1, 2, 3, 4 or 5;

[0013] p is selected from 0, 1, 2, 3, 4 or 5.

[0014] In an alternative embodiment, R is selected from the following groups, which are unsubstituted or optionally substituted by one, two or more R 2 substituted C 1-6 alkyl, phenyl;

[0015] In an alternative embodiment, R is selected from C 1-6 alkyl,

[0016] In an alternative embodiment, R is selected from n-butyl, t-butyl,

[0017] In an alternative embodiment, each R 1 is the same or different and independently of each other selected from H, halogen, C 1-6 alkyl, C 1-6 alkoxy;

[0018] In an alternative embodiment, each R 1 is the same or different and independently of each other selected from H, F, methoxy.

[0019] In an alternative embodiment, each R 2 is the same or different and independently of each other selected from H, halogen, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, halogenated C 1-6 alkoxy, phenyl;

[0020] In an alternative embodiment, each R 2 is the same or different and independently of each other selected from H, F, CI, methoxy, trifluoromethyl, phenyl.

[0021] In an alternative embodiment, each R 3 is the same or different and independently of each other selected from H, halogen, OH, CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkyl-phenyl-S(O)2-NH-, phenyl-C 1-6 alkoxy;

[0022] In an alternative embodiment, each R 3 is the same or different and independently of each other selected from H, F, CI, Br, OH, CN, methyl, methoxy, 4-methylphenylsulfonyl-NH-, benzyloxy.

[0023] In an alternative embodiment, each R 4They are either the same or different, and are independently selected from H and methyl.

[0024] In an optional implementation, each R a R b R c R d They may be the same or different, and are independently selected from H, halogens, OH, CN, NO2, and C. 1-6 Alkyl, C 1-6 Alkoxy, phenyl, 5-6 membered heteroaryl, 3-6 membered heterocyclic, C 1-6 Alkyl-phenyl-S(O)2-;

[0025] In an optional implementation, each R a R b R c R d They may be the same or different, and are independently selected from H, halogen, OH, CN, NO2, methyl, methoxy, phenyl, and 4-methylbenzenesulfonyl.

[0026] In an optional embodiment, the compounds represented by formula (I) and / or formula (II) are selected from the structures shown below:

[0027]

[0028] Among them, R 1 R 2 R 3 R 4 They each possess the definitions described above independently.

[0029] In an optional embodiment, the compound represented by formula (I) is selected from the following structures:

[0030]

[0031] In an optional embodiment, the compound represented by formula (II) is selected from the structures shown below:

[0032] Preparation method

[0033] The present invention also provides a method for preparing compounds of formula (I) and / or formula (II) or pharmaceutically acceptable salts thereof, comprising the following steps: reacting a first substrate compound a and a second substrate compound b in the presence of a catalyst to obtain compounds of formula (I) and / or formula (II);

[0034]

[0035] Among them, R, R 1 R 3 R 4, m, p are independently of each other defined as described above; L is selected from C 1-6 alkoxy, for example methoxy;

[0036] In an alternative embodiment, the catalyst is selected from catalyst 1 or catalyst 2; wherein catalyst 1 is selected from rac-BPA TsOH, CF3CO2H, CF3SO3H, Cu(OTf)2; catalyst 2 is selected from AlCl3; when the catalyst is catalyst 1, the reaction produces compounds of formula (I) and (II); when the catalyst is catalyst 2, the reaction produces compounds of formula (II);

[0037] In an alternative embodiment, the molar ratio of compound a, compound b and catalyst is selected from (0.2-5):1:(0.01-0.1), for example (0.9-3):1:(0.02-0.0.05), (0.2-0.8):1:(0.01-0.1), for example 1.5:1:0.025, 0.5:1:0.025;

[0038] In an alternative embodiment, the molar ratio of compound a, compound b and catalyst 1 is selected from (0.9-3):1:(0.02-0.0.05), for example 1.5:1:0.025;

[0039] In an alternative embodiment, the molar ratio of compound a, compound b and catalyst 2 is selected from (0.2-0.8):1:(0.01-0.1), for example 0.5:1:0.025;

[0040] In an alternative embodiment, the reaction is carried out in the presence of an organic solvent selected from at least one of DCM, toluene, methanol, ethanol, THF, 2-methyltetrahydrofuran, 1,4-dioxane, acetonitrile, DMF.

[0041] In an alternative embodiment, the temperature of the reaction is selected from 10-40℃, for example 20℃, 25℃, 30℃, 35℃; the time of the reaction is selected from 0.1-6h, for example 0.5h, 1h, 2h, 3h, 4h, 5h.

[0042] The present application also provides the use of the compound of formula (II) as raw material in further preparation of its derivatives.

[0043] The present application also provides a method for further preparation of derivatives of the compound of formula (II) using the compound of formula (II) as raw material, comprising the following scheme one, scheme two or scheme three:

[0044] Scheme one: the compound of formula (II) is reacted in the presence of a reducing agent to obtain a compound of formula (II-a);

[0045]

[0046] wherein R, R 1 , R 3 , R 4 , m, p independently of each other have the definitions described above;

[0047] In an alternative embodiment, the reducing agent is selected from at least one of sodium borohydride, potassium borohydride, sodium borohydride acetate, sodium cyanoborohydride;

[0048] Scheme II: the compound shown as formula (II-a) is reacted with acetic anhydride to obtain the compound shown as formula (II-b);

[0049]

[0050] wherein R, R 1 , R 3 , R 4 , m, p independently of each other have the definitions described above;

[0051] Scheme III: the compound shown as formula (II) is reacted with hydroxylamine hydrochloride to obtain the compound shown as formula (II-c);

[0052]

[0053] wherein R, R 1 , R 3 , R 4 , m, p independently of each other have the definitions described above.

[0054] The present application also provides a compound shown as follows:

[0055]

[0056] wherein R, R 1 , R 3 , R 4 , m, p independently of each other have the definitions described above.

[0057] The present application also provides a pharmaceutical composition comprising at least one of the compound shown as formula (I) and / or formula (II) or a pharmaceutically acceptable salt thereof.

[0058] In an alternative embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant, such as a carrier or an excipient.

[0059] Pharmaceutical use

[0060] The application also provides use of at least one of the compounds shown in the formula (I) and / or formula (II) or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing and / or treating cancer.

[0061] In an alternative embodiment, the cancer is breast cancer.

[0062] Beneficial effects

[0063] The present application provides indole heterocyclic compounds shown in the formula (I) and / or formula (II) or a pharmaceutically acceptable salt thereof, pharmaceutical compositions, and synthetic methods and applications thereof. The compounds of the present application have good anti-tumor activity, and the synthetic method of the compounds is simple. By selecting different types of catalysts and regulating the ratio of substrates, the compounds shown in the formula (I) and / or formula (II) can be selectively synthesized. The compounds can be further used to prepare reduced, acylated or ketoxime derived compounds. BRIEF DESCRIPTION OF DRAWINGS

[0064] Figure 1 Results of MTT method for determining the cytotoxic effect of each compound on MCF-7 cells and MCF-10A;

[0065] Figure 2 Results of flow cytometry for detecting apoptosis of MCF-7 cells treated with compounds 2-4ak;

[0066] Figure 3 Results of DAPI staining method for analyzing MCF-7 cells treated with compounds 2-4ak;

[0067] Figure 4 Results of transmission electron microscopy for observing the ultrastructure of MCF-7 cells treated with compounds 2-4ak;

[0068] Figure 5 Results of Western blotting for detecting the expression of different proteins in MCF-7 cells treated with compounds 2-4ak;

[0069] Figure 6 Results of analysis of MCF-7 xenograft tumor model treated with compounds 2-4ak;

[0070] Figure 7 Results of H&E, TUNEL and immunofluorescence staining of tumor tissues of MCF-7 tumor-bearing mice treated with compounds 2-4ak;

[0071] Figure 8 Results of H&E staining of heart, liver, spleen, lung and kidney tissues of mice treated with compounds 2-4ak.

[0072] Definitions and explanations of terms

[0073] Unless otherwise indicated, the definitions of groups and terms in the present application specification and claims, including the definitions of examples, illustrative examples, preferred definitions, definitions recited in tables, definitions of specific compounds in examples, etc., can be combined and incorporated with each other in any manner. The group definitions and compound structures after such combination and incorporation should be understood as within the scope recited in the present application specification and / or claims.

[0074] The term "alkyl" denotes straight-chain or branched saturated hydrocarbon groups, preferably straight-chain or branched saturated hydrocarbon groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms (C 1-10 alkyl). The alkyl groups include C 1-3 alkyl, C 1-6 alkyl, C 3-6 alkyl, C 1-10 alkyl, etc. The "C 1-10 alkyl" denotes straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, the "C 1-8 alkyl" denotes straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, the "C 1-6 alkyl" denotes straight-chain and branched alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl groups are, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1 -methylbutyl, 1 -ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1 -dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1 -methylpentyl, 2-ethylbutyl, 1 -ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1 -dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, and the like or their isomers.

[0075] The term "cycloalkyl" denotes saturated, monovalent monocyclic, bicyclic (e.g. fused, bridged, spirocyclic) or tricyclic hydrocarbon rings, preferably having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms (C 3-10 cycloalkyl). The cycloalkyl groups include C 3-8 cycloalkyl, C 3-5 cycloalkyl, C 6-8 cycloalkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, C6cycloalkyl, C 3-10 cycloalkyl, etc. The C 3-10Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or bicyclic hydrocarbon groups, such as borneolyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl; or tricyclic hydrocarbon groups, such as adamantyl.

[0076] The term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic or polycyclic substituent comprising 3 to 10 ring atoms, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen, and sulfur, wherein the sulfur may optionally be oxidized (i.e., forming a sulfoxide or sulfone), but excluding the -OO-, -OS-, or -SS- ring moiety, and the remaining ring atoms are carbon. Preferably, it comprises 3 to 8 (e.g., 3, 4, 5, 6, 7, or 8) ring atoms, wherein 1 to 4 (e.g., 1, 2, 3, and 4) are heteroatoms; more preferably, it comprises 3 to 6 ring atoms, wherein 1 to 3 are heteroatoms; most preferably, it comprises 5 or 6 ring atoms, wherein 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, tetrahydropyranyl, 1,2,3,6-tetrahydropyridyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc.

[0077] The term "aryl" should be understood to preferably refer to a monocyclic, bicyclic, or tricyclic hydrocarbon ring having 6, 7, 8, 9, or 10 carbon atoms and possessing monovalent aromatic or partially aromatic properties ("C"). 6-10 Aryl), particularly a ring with 6 carbon atoms (“C6 aryl”), such as phenyl; or biphenyl, or a ring with 9 carbon atoms (“C9 aryl”), such as indenyl or indenyl, or a ring with 10 carbon atoms (“C9 aryl”). 10 Aryl), such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl. When the C 6-10 When the aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, it can be ortho, para, or meta substituted.

[0078] The term "heteroaryl" means a monovalent or polyvalent monocyclic, bicyclic or tricyclic aromatic ring system having 5, 6, 7, 8, 9 or 10 ring atoms, and of which the ring atoms include 1-5 heteroatoms independently selected from N, O and S, which bicyclic and tricyclic aromatic ring systems can be fused, spiro or bridged (5-10 membered heteroaryl). The 5-10 membered heteroaryl has 1-5, preferably 1-3 heteroatoms. Additionally, in each instance the 5-10 membered heteroaryl can be benzo-fused. The 5-10 membered heteroaryl includes 5-6 membered heteroaryl, 9-10 membered heteroaryl. Examples of heteroaryl include, but are not limited to: 5-membered rings such as oxazolyl, pyrazolyl, thienyl, thiazolyl, triazolyl, imidazolyl, and the like; 6-membered rings such as pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, and the like. The heteroaryl can be bicyclic, including but not limited to: 5,5 membered rings such as tetrahydrocyclopentapyrazole; 5,6 membered rings such as tetrahydroindole, tetrahydropyrazolopyridine, tetrahydroimidazopyridine, tetrahydrobenzisoxazole, tetrahydrobenzoxazole, tetrahydrobenzothiazole, tetrahydrobenzisothiazole, dihydrofuropyrazole, tetrahydrobenzofuran, dihydrobenzofuran, tetrahydrobenzothiophene; 6,6 membered rings such as tetrahydroquinoline; 5,7 membered rings such as tetrahydrocycloheptylthiazole, tetrahydrocycloheptylfuran. When the 5-10 membered heteroaryl is substituted, it can be mono- or poly-substituted. Also, there is no limitation on the substitution site, for example, the hydrogen attached to a carbon atom on the heteroaryl ring can be substituted, or the hydrogen attached to a heteroatom on the heteroaryl ring can be substituted.

[0079] The term "pharmaceutically acceptable" as used herein means a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present application, and is relatively nontoxic, i.e., the material is not biologically or otherwise undesirable, with respect to the

[0080] As will be understood by those skilled in the art, the compounds of the present disclosure can exist in various pharmaceutically acceptable salt forms. If the compounds have a basic center, they can form acid addition salts; if the compounds have an acidic center, they can form base addition salts; if the compounds contain both an acidic center (such as a carboxyl group) and a basic center (such as an amino group), they can also form inner salts.

[0081] The compounds defined or disclosed herein include all stereoisomers and optical isomers and racemates thereof, where such isomers exist. Unless otherwise specified, all chiral (enantiomeric and diastereomeric) and racemic forms are within the scope of the application. The compounds of the present application can be isolated in optically active or racemic forms. The optically active forms can be prepared by separation from a racemic mixture or by stereospecific synthesis from an optically active starting material. All processes used to prepare compounds of the present application and intermediates used therein are considered to be part of the present application. When enantiomeric or diastereomeric products are prepared, they can be separated by ordinary DETAILED DESCRIPTION

[0082] In order to make the purposes, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. If specific conditions are not indicated in the embodiments, the conditions are implemented according to conventional conditions or conditions recommended by manufacturers. If manufacturers of reagents or instruments are not indicated, the reagents and instruments are conventional products that can be purchased in the market.

[0083] The features and properties of the present application are further described in detail below in combination with the embodiments.

[0084] Embodiment 1

[0085] This embodiment provides a synthesis method of two indole heterocyclic compounds 2-3aa and 2-4aa, and the synthesis reaction formula is as follows:

[0086]

[0087] Addition reaction: 0.3 mmol of the first substrate 2-1a, 0.2 mmol of the second substrate 2-2a and 2.5 mol% of rac-BPA (catalyst) were placed in 2.0 ml of solvent MeCN, and the reaction was carried out at room temperature for 1 h to obtain the reaction products 2-3aa and 2-4aa, with the yield being 84% and 7% respectively.

[0088] Addition ring-opening reaction: 0.2 mmol of the first substrate 2-1a, 0.4 mmol of the second substrate 2-2a and 2.5 mol% of AlCl3 (catalyst) were placed in 2.0 ml of solvent MeCN, and the reaction was carried out at room temperature for 1 h to obtain the reaction product 2-4aa, with the yield being 90%.

[0089] The structural formula of 2-3aa is as follows,

[0090] Chemical name: 3-(3-phenyl-1H-isochromen-1-yl)-1H-indole [3-(3-Phenyl-1H- isochromen-1-yl)-1H-indole] (2-3aa), yellow solid (54.2 mg, yield 84%). mp 135.8-137.6 °C; 1 H NMR (500 MHz, CDC13) δ 7.99 (s, 1H), 7.90 (d, J = 7.7 Hz, 1H), 7.72-7.67 (m, 2H), 7.35-7.28 (m, 5H), 7.25-7.12 (m, 4H), 7.00 (d, J = 7.5 Hz, 1H), 6.84 (s, 1H), 6.72 (s, 1H), 6.54 (s, 1H); 13 C NMR (125 MHz, CDC13) δ 152.7, 136.7, 134.9, 132.1, 130.4, 128.7, 128.3, 128.2, 126.5, 126.5, 125.5, 125.2, 125.0, 123.8, 122.5, 120.2, 120.1, 115.3, 111.4, 101.0, 74.1; HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 18 NO, 323.1310; found: 323.1312.

[0091] wherein the structural formula of 2-4aa is,

[0092] Chemical name: 2-(2-(di(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-one [2-(2-(Di(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-one] (4aa), white solid (63.7 mg, yield 72%). mp 154.4-156.2 °C; 1 H NMR (500 MHz, CDC13) δ 7.89 (s, 2H), 7.78 (d, J = 7.3 Hz, 2H), 7.47 (t, J = 7.4 Hz, 1H), 7.35 (d, J = 7.9 Hz, 2H), 7.34-7.26 (m, 4H), 7.23-7.18 (m, 2H), 7.18-7.08 (m, 4H), 6.98 (t, J = 7.5 Hz, 2H), 6.49 (s, 2H), 5.92 (s, 1H), 4.32 (s, 2H); 13C NMR (125 MHz, CDC13) δ 198.7, 142.5, 137.0, 136.8, 133.6, 133.1, 131.1, 128.9, 128.5, 128.4, 127.2, 127.1, 126.6, 124.3, 122.1, 120.1, 119.5, 118.7, 111.1, 42.7, 36.9; HRMS (ESI) m / z: [M + H] + calcd. for C 31 H 25 N2O, 441.1961; found: 441.1964.

[0093] Example 2

[0094] Example 2 is substantially the same as Example 1, the only difference is that the catalyst, solvent or reaction time used in the reaction of the first substrate and the second substrate. When the catalyst is rac-BPA, other conditions remain unchanged, use different solvents: DCM, toluene, MeOH, THF, wherein the reaction time of THF > 48 h; when the solvent is MeCN, other conditions remain unchanged, use different catalysts: BINOL, TsOH, CF3CO2H, CF3SO3H, AICI3, Cu(OTf)2. The condition screening is shown in Table 1.

[0095] Table 1 Screening of reaction conditions for isobenzopyran and indole

[0096]

[0097] [a] Reaction conditions: 2-1a (0.2 mmol), 2-2a (0.2 mmol), isolated yield. [b] Reaction conditions: 2-1a (0.3 mmol), 2-2a (0.2 mmol). [c] Reaction conditions: 2-1a (0.2 mmol), 2-2a (0.4 mmol).

[0098] Example 3

[0099] According to the reaction conditions of the addition reaction of Example 1, different indole heterocyclic compounds are obtained by changing the second substrate, and the specific reaction formula is as follows:

[0100]

[0101] The structural formula of the obtained indole heterocyclic compound and its yield are as follows:

[0102]

[0103] The structural formula of compound 2-3ab is

[0104] The chemical name is: 1-methyl-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole [1-methyl-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole] (2-3ab). Brown solid; 50.6 mg, yield 75%, mp 136.2-138.1°C; 1 H NMR (500 MHz, CDCI3) δ 7.92 (d, J = 7.8 Hz, 1 H), 7.70 (d, J = 6.9 Hz, 2H), 7.37-7.26 (m, 6H), 7.26-7.14 (m, 3H), 7.04 (d, J = 7.4 Hz, 1 H), 6.74 (s, 1 H), 6.72 (s, 1 H), 6.53 (s, 1 H), 3.69 (s, 3H); 13 C NMR (125 MHz, CDCI3) δ 152.6, 137.5, 135.0, 132.1, 130.5, 129.6, 128.7, 128.3, 128.2, 127.1, 126.4, 125.5, 125.2, 123.7, 122.0, 120.2, 119.7, 113.8, 109.6, 100.8, 74.0, 32.9; HRMS (ESI) m / z: [M + H] + calcd. for C 24 H 20 NO, 338.1539; found: 338.1537.

[0105] The structural formula of compound 2-3ab is

[0106] The chemical name is: 1-methyl-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole [1-methyl-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole] (2-3ab). Brown solid; 50.6 mg, yield 75%, mp 136.2-138.1°C; 1H NMR (500 MHz, CDC13) δ 7.91 (s, 1H), 7.81-7.75 (m, 2H), 7.51 (d, J = 7.9 Hz, 1H), 7.40-7.28 (m, 5H), 7.24 (d, J = 7.5 Hz, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.10-7.05 (m, 2H), 6.79 (d, J = 7.5 Hz, 1H), 6.65 (s, 1H), 6.56 (s, 1H), 2.39 (s, 3H); 13 C NMR (125 MHz, CDC13) δ 154.4, 135.5, 134.8, 134.6, 132.9, 130.9, 128.8, 128.4, 128.1, 127.9, 126.6, 125.3, 125.1, 123.5, 121.4, 120.2, 119.8, 110.5, 109.4, 101.1, 74.7, 12.3; HRMS (ESI) m / z: [M + H] + calcd. for C 24 H 20 NO, 338.1539; found: 338.1537.

[0107] The structural formula of compound 2-3ad is,

[0108] Chemical name: 4-methyl-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole (2-3ad). Yellow solid; 56.7 mg, yield 84%, mp 193.6-197.3 °C; 1 H NMR (500 MHz, DMSO-d6) δ 11.07 (s, 1H), 7.62 (d, J = 7.5 Hz, 2H), 7.34-7.25 (m, 6H), 7.21 (d, J = 8.1 Hz, 1H), 7.16 (t, J = 7.3 Hz, 1H), 7.00 (t, J = 7.6 Hz, 2H), 6.89 (s, 1H), 6.83 (d, J = 7.2 Hz, 1H), 6.75 (s, 1H), 2.73 (s, 3H); 13C NMR (125MHz, DMSO-d6) δ150.2,137.2,134.5,131.4,131.1,129.8,128.7,128.4,128.0,127.0,126. 5,125.1,124.9,124.5,123.6,121.6,120.8,113.4,109.6,101.0,73.4,20.5; HRMS(ESI)m / z:[M+H] + calcd.for C 24 H 20 NO. 338.1539; found: 338.1537.

[0109] The structural formula of compound 2-3ae is,

[0110]

[0111] Chemical name: 6-methyl-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole [6-methyl-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole](2-3ae). Red solid; 44.9 mg, yield 67%, mp 111.9-116.4℃; 1 H NMR (500MHz, CDCl) 3 )δ7.92(s,1H),7.73(d,J=8.1Hz,1H),7.68-7.65(m,2H),7.31-7.24(m,4H),7.18(d,J=6.7Hz,1H),7.18-7.14(m,1H),7.12(td,J= 7.5,1.3Hz,1H),7.00(dd,J=8.1,1.0Hz,1H),6.97(d,J=7.5Hz,1H),6.83(d,J=2.4Hz,1H),6.65(s,1H),6.50(s,1H),2.47(s,3H); 13 C NMR (125MHz, CDCl3) δ152.8,137.2,134.9,132.4,132.2,130.5,128.7,128.3,128.2,126.5,125.5 ,125.2,124.4,124.4,123.7,122.0,120.0,115.3,111.3,100.9,74.3,21.9; HRMS(ESI)m / z:[M+H] + calcd.for C 24 H 20NO, 338.1539; found: 338.1538.

[0112] The structural formula of compound 2-3af is,

[0113] Chemical name: 4-Methyl-N-(3-(3-phenyl-1H-isochromen-1-yl)-1H-indol-6- yl)benzenesulfonamide (2-3af). Brown solid; 73.9 mg, yield 75%, mp 173.4-175.4 °C; 1 H NMR (500 MHz, DMSO-d6) δ 11.00 (s, 1H), 9.98 (s, 1H), 7.64-7.59 (m, 4H), 7.53-7.47 (m, 1H), 7.33-7.26 (m, 6H), 7.23 (d, J = 7.4 Hz, 1H), 7.18 (d, J = 8.0 Hz, 1H), 7.13 (td, J = 7.4, 1.4 Hz, 1H), 6.94 (d, J = 7.5 Hz, 1H), 6.86-6.77 (m, 2H), 6.71 (s, 1H), 6.65 (s, 1H), 2.28 (s, 3H); 13 C NMR (125 MHz, DMSO-d6) δ 151.2, 142.9, 136.8, 136.7, 134.3, 131.9, 131.4, 130.2, 129.5, 128.7, 128.3, 128.1, 126.7, 126.4, 125.8, 124.9, 124.8, 123.7, 123.1, 119.7, 114.2, 113.7, 104.3, 100.8, 73.3, 21.0; HRMS (ESI) m / z: [M+H] + calcd. for C 30 H 25 N2O3S, 493.1580; found: 493.1580.

[0114] The structural formula of compound 2-3ah is,

[0115] Chemical name: 4-((3-Phenyl-1H-isochromen-1-yl)oxy)-1H-indole (2-3ah). Yellow solid; 47.8 mg, yield 70%, mp 150.1-153.2 °C;1 H NMR (500 MHz, CDC13) δ 8.15 (s, 1H), 7.75-7.67 (m, 2H), 7.39-7.27 (m, 5H), 7.22 (d, J = 7.0 Hz, 1H), 7.17-7.08 (m, 2H), 6.95 (d, J = 8.1 Hz, 1H), 6.84-6.77 (m, 2H), 6.64 (d, J = 7.7 Hz, 1H), 6.62 (s, 1H), 6.59 (s, 1H); 13 C NMR (125 MHz, CDC13) δ 152.0, 150.1, 139.1, 133.9, 131.5, 130.5, 129.2, 128.8, 128.6, 127.2, 125.2, 125.2, 124.5, 124.2, 123.7, 115.9, 112.7, 106.1, 103.8, 103.4, 75.9; HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 18 NO2, 340.1332; found: 340.1331.

[0116] The structural formula of compound 2-3ai is,

[0117] Chemical name: 4-Fluoro-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole (2-3ah). Yellow solid; 51.9 mg, yield 76%, mp 149.0-150.3 °C; 1 H NMR (500 MHz, CDC13) δ 8.15 (s, 1H), 7.75-7.67 (m, 2H), 7.39-7.27 (m, 5H), 7.22 (d, J = 7.0 Hz, 1H), 7.17-7.08 (m, 2H), 6.95 (d, J = 8.1 Hz, 1H), 6.84-6.77 (m, 2H), 6.64 (d, J = 7.7 Hz, 1H), 6.62 (s, 1H), 6.59 (s, 1H); 13C NMR (125 MHz, CDCI3) δ 157.1 (d, J = 247.4 Hz), 151.8, 139.2 (d, J = 11.6 Hz), 134.9, 131.8, 130.6, 128.7, 128.3, 128.2, 126.5, 125.4, 125.3, 125.2, 123.8, 123.1 (d, J = 7.8 Hz), 115.5 (d, J = 20.9 Hz), 114.2, 107.5 (d, J = 3.7 Hz), 105.6 (d, J = 19.6 Hz), 100.7, 73.4; 19 F NMR (471 MHz, CDCI3) δ -115.72 - -125.95 (m, IF); HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 17 F NO, 342.1289; found: 342.1288.

[0118] The structural formula of compound 2-3aj is,

[0119] Chemical name: 4-bromo-3-(3-phenyl-1 H-isochromen-1 -yl)-1 H-indole [4-Bromo-3-(3-phenyl-1 H-isochromen-1 -yl)-1 H-indole] (2-3aj). Brown solid; 68.7 mg, yield 85%, mp 175.2-177.2 °C; 1 H NMR (500 MHz, CDCI3) δ 8.09 (s, 1 H), 7.74-7.69 (m, 2H), 7.40 (d, J = 7.6 Hz, 1 H), 7.36 (s, 1 H), 7.31 -7.24 (m, 4H), 7.26-7.20 (m, 1 H), 7.20-7.13 (m, 2H), 7.08-7.01 (m, 2H), 6.72 (s, 1 H), 6.49 (s, 1 H); 13 C NMR (125 MHz, CDCI3) δ 151.5, 137.8, 135.0, 132.0, 130.6, 128.7, 128.3, 128.2, 127.5, 126.6, 125.6, 125.4, 125.2, 125.0, 123.7, 123.3, 115.8, 114.2, 110.7, 100.7, 71.9; HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 17BrNO, 402.0488; found: 402.0487.

[0120] The structural formula of compound 2-3ak is,

[0121] Chemical name: 3-(3-phenyl-1H-isochromen-1-yl)-1H-indole-4-carbonitrile [3-(3-Phenyl-1H-isochromen-1-yl)-1H-indole-4-carbonitrile] (2-3ak). Brown solid; 44.1 mg, yield 63%, mp 181.5-183.9 °C; 1 H NMR (500 MHz, CDC13) δ 8.54 (s, 1H), 7.72-7.66 (m, 2H), 7.57 (dd, J = 7.4, 0.9 Hz, 1H), 7.53 (dd, J = 8.2, 0.9 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 7.27-7.19 (m, 4H), 7.19-7.15 (m, 2H), 7.14-7.12 (m, 1H), 7.01 (d, J = 7.5 Hz, 1H), 6.78 (s, 1H), 6.47 (s, 1H); 13 C NMR (125 MHz, CDC13) δ 151.3, 136.9, 134.8, 131.9, 129.7, 128.7, 128.6, 128.4, 128.3, 127.1, 126.6, 126.1, 125.2, 125.2, 123.9, 122.0, 119.3, 116.6, 115.3, 102.5, 100.7, 71.8; HRMS (ESI) m / z: [M + H] + calcd. for C 24 H 17 N2O, 349.1335; found: 349.1334.

[0122] The structural formula of compound 2-3al is,

[0123] Chemical name: 3-(3-phenyl-1H-isochromen-1-yl)-1H-indole-4-carbonitrile [3-(3-Phenyl-1H-isochromen-1-yl)-1H-indole-4-carbonitrile] (2-3ak). Brown solid; 44.1 mg, yield 63%, mp 181.5-183.9 °C; 1H NMR (500 MHz, CDC13) δ 8.02 (s, 1H), 7.85 (d, J = 1.9 Hz, 1H), 7.72-7.65 (m, 2H), 7.35-7.27 (m, 4H), 7.23-7.14 (m, 4H), 6.96 (d, J = 7.5 Hz, 1H), 6.83 (d, J = 2.6 Hz, 1H), 6.63 (s, 1H), 6.53 (s, 1H); 13 C NMR (125 MHz, CDC13) δ 152.5, 135.0, 134.7, 132.0, 130.0, 128.8, 128.4, 127.5, 126.6, 126.3, 125.8, 125.4, 125.1, 123.9, 122.9, 119.7, 115.1, 112.5, 101.1, 73.9; HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 17 ClNO, 358.0993; found: 358.0992.

[0124] The chemical name of compound 2-3am is,

[0125] The chemical name is: 6-Fluoro-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole (2-3am). White solid; 39.1 mg, yield 57%; mp 136.5-137.2 °C; 1 H NMR (500 MHz, CDC13) δ 8.02 (s, 1H), 7.85 (d, J = 1.9 Hz, 1H), 7.72-7.65 (m, 2H), 7.35-7.27 (m, 4H), 7.23-7.14 (m, 4H), 6.96 (d, J = 7.5 Hz, 1H), 6.83 (d, J = 2.6 Hz, 1H), 6.63 (s, 1H), 6.53 (s, 1H); 13C NMR (125 MHz, CDC13) δ 160.2 (d, J = 238.2 Hz), 152.6, 136.7 (d, J = 12.4 Hz), 134.8, 132.1, 130.2, 128.8, 128.4, 128.3, 126.5, 125.5, 125.3 (d, J = 3.3 Hz), 125.2, 123.8, 123.1, 121.1 (d, J = 10.1 Hz), 115.6, 109.0 (d, J = 24.4 Hz), 101.0, 97.8 (d, J = 26.1 Hz), 74.0; 19 F NMR (471 MHz, CDC13) δ -120.62 (td, J = 9.6, 5.3 Hz, IF); HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 17 FNO, 342.1289; found: 342.1289.

[0126] The structural formula of compound 2-3an is,

[0127] Chemical name: 7-Fluoro-3-(3-phenyl-1H-isochromen-1-yl)-1H-indole (2-3an). White solid; 62.9 mg, yield 92%; mp 173.4-175.0 °C; 1 H NMR (500 MHz, CDC13) δ 8.22 (s, 1H), 7.69-7.65 (m, 2H), 7.63 (d, J = 8.0 Hz, 1H), 7.34-7.25 (m, 4H), 7.20 (d, J = 7.4 Hz, 1H), 7.15 (td, J = 7.5, 1.2 Hz, 1H), 7.08 (td, J = 7.9, 4.8 Hz, 1H), 6.98 (d, J = 7.5 Hz, 1H), 6.94 (dd, J = 11.2, 7.9 Hz, 1H), 6.89 (d, J = 2.4 Hz, 1H), 6.68 (s, 1H), 6.51 (s, 1H); 13C NMR (125 MHz, CDC13) δ 152.5, 149.7 (d, J = 244.0 Hz), 134.8, 132.0, 130.2 (d, J = 5.0 Hz), 130.1, 128.8, 128.4, 128.4, 126.6, 125.6, 125.5, 125.1, 125.1 (d, J = 13.1 Hz), 123.9, 120.5, 120.5, 116.4 (d, J = 2.1 Hz), 116.1 (d, J = 3.6 Hz), 107.4 (d, J = 16.0 Hz), 101.0, 73.9; 19 F NMR (471 MHz, CDC13) δ -135.17 - -135.20 (m, IF); HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 17 FNO, 342.1289; found: 342.1289.

[0128] Example 4

[0129] According to the reaction conditions of the addition reaction of Example 1, different indole heterocyclic compounds were obtained by changing the first substrate, and the specific reaction formula is as follows:

[0130]

[0131] The structural formula of the obtained polyaryl-substituted indole heterocyclic compound and its yield are as follows:

[0132]

[0133] The structural formula of compound 2-3ba is,

[0134] Chemical name: 3-(3-(3-chlorophenyl)-1H-isochromen-1-yl)-1H-indole [3-(3-(3-chlorophenyl)-1H-isochromen-1-yl)-1H-indole] (2-3ba). Red solid, 57.7 mg, yield 81%, mp 118.3-120.4 °C; 1 H NMR (600 MHz, CDC13) δ 7.90 (s, 1H), 7.81 (d, J = 7.4 Hz, 1H), 7.61 (s, 1H), 7.47 (d, J = 7.5 Hz, 1H), 7.27 (d, J = 7.9 Hz, 2H), 7.20-7.07 (m, 6H), 6.93 (d, J = 7.4 Hz, 1H), 6.74 (s, 1H), 6.63 (s, 1H), 6.45 (s, 1H).13 C NMR (150 MHz, CDC13) δ 151.2, 136.7, 136.6, 134.3, 131.6, 130.5, 129.5, 128.5, 128.3, 126.9, 126.4, 125.5, 125.3, 125.1, 124.0, 123.5, 122.6, 120.2, 120.2, 115.1, 111.5, 102.0, 74.3; HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 17 ClNO, 358.0993; found: 358.0991.

[0135] The structural formula of compound 2-3ca is,

[0136] Chemical name: 3-(3-(3-methoxyphenyl)-1H-isochromen-1-yl)-1H-indole [3-(3-(3-Methoxyphenyl)-1H-isochromen-1-yl)-1H-indole] (2-3ca). White solid, 35.4 mg, yield 50%, mp 212.7-213.5 °C; 1 H NMR (500 MHz, DMSO-d6) δ 11.06 (s, 1H), 7.74 (d, J = 7.9 Hz, 1H), 7.38 (d, J = 8.1 Hz, 1H), 7.31 (t, J = 7.4 Hz, 1H), 7.28 - 7.21 (m, 3H), 7.21 - 7.15 (m, 2H), 7.13 - 7.08 (m, 1H), 7.05 - 7.00 (m, 2H), 6.84 (dt, J = 7.3, 2.1 Hz, 1H), 6.82 (d, J = 2.4 Hz, 1H), 6.75 (d, J = 5.8 Hz, 2H), 3.70 (s, 3H); 13 C NMR (125 MHz, DMSO-d6) δ 159.2, 150.9, 136.7, 135.9, 131.4, 130.3, 129.4, 128.1, 126.5, 126.0, 125.7, 125.0, 123.8, 121.5, 119.4, 119.1, 117.2, 114.5, 113.8, 111.8, 110.1, 101.2, 73.3, 55.1; HRMS (ESI) m / z: [M + H] + calcd. for C 24 H 20 NO2, 354.1489; found: 354.1488.

[0137] The chemical name of compound 2-3da is:

[0138] The chemical name of compound 2-3da is: 1 H NMR (500 MHz, CDC13) δ 8.04 (s, 1H), 7.88 (d, J = 7.9 Hz, 1H), 7.63-7.58 (m, 2H), 7.36 (d, J = 8.1 Hz, 1H), 7.30-7.26 (m, 1H), 7.25-7.21 (m, 1H), 7.20-7.16 (m, 2H), 7.11 (td, J = 7.5, 1.2 Hz, 1H), 6.97 (d, J = 7.4 Hz, 1H), 6.86 (d, J = 2.4 Hz, 1H), 6.84-6.79 (m, 2H), 6.67 (s, 1H), 6.39 (s, 1H), 3.78 (s, 3H); 13 C NMR (125 MHz, CDC13) δ 160.2, 152.7, 136.7, 132.5, 130.2, 128.2, 127.6, 127.1, 126.6, 126.1, 125.2, 125.0, 123.5, 122.5, 120.3, 120.1, 115.5, 113.7, 111.4, 99.4, 74.1, 55.4; HRMS (ESI) m / z: [M + H] + calcd. for C 24 H 20 NO2, 354.1489; found: 354.1488.

[0139] The chemical name of compound 2-3da is:

[0140] The chemical name of compound 2-3da is: 1H NMR (500 MHz, CDC13) δ 8.72 (s, 1H), 7.85 (d, J = 7.8 Hz, 1H), 7.73 (d, J = 8.2 Hz, 2H), 7.49 (d, J = 8.3 Hz, 2H), 7.36 (d, J = 8.0 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 7.24-7.17 (m, 2H), 7.18-7.13 (m, 2H), 7.00 (d, J = 7.5 Hz, 1H), 6.81 (d, J = 2.6 Hz, 1H), 6.71 (s, 1H), 6.56 (s, 1H); 13 C NMR (125 MHz, CDC13) δ 150.9, 138.3, 136.8, 131.4, 130.5, 130.0 (q, J = 32.2 Hz), 128.3, 127.1, 126.4, 125.4, 125.3, 125.3, 125.2 (q, J = 3.8 Hz), 124.2 (q, J = 272.0 Hz), 124.1, 122.4, 120.0, 119.9, 114.9, 111.6, 102.7, 74.2; 19 F NMR (471 MHz, CDC13) δ -62.6; HRMS (ESI) m / z: [M + H] + calcd. for C 24 H 17 F3NO, 392.1257; found: 392.1256.

[0141] The chemical name of compound 2-3ja is,

[0142] The chemical name is: 3-(6-Fluoro-3-phenyl-1H-isochromen-1-yl)-1H-indole (2-3ja). Yellow solid, 74.3 mg, yield 72%, mp 156.3-158.0 °C; 1 H NMR (500 MHz, CDC13) δ 8.72 (s, 1H), 7.85 (d, J = 7.8 Hz, 1H), 7.73 (d, J = 8.2 Hz, 2H), 7.49 (d, J = 8.3 Hz, 2H), 7.36 (d, J = 8.0 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 7.24-7.17 (m, 2H), 7.18-7.13 (m, 2H), 7.00 (d, J = 7.5 Hz, 1H), 6.81 (d, J = 2.6 Hz, 1H), 6.71 (s, 1H), 6.56 (s, 1H); 13C NMR (125 MHz, CDC13) δ 162.9 (d, J = 244.3 Hz), 153.8, 136.7, 134.4, 134.3 (d, J = 9.0 Hz), 129.1, 128.4, 126.8 (d, J = 8.7 Hz), 126.4, 126.0 (d, J = 2.8 Hz), 125.7, 125.0, 122.6, 120.3, 115.2, 112.8 (d, J = 22.0 Hz), 111.5, 110.2 (d, J = 22.5 Hz), 100.3, 100.3, 74.0; 19 F NMR (471 MHz, CDC13) δ -114.94 (q, J = 8.6 Hz); HRMS (ESI) m / z: [M + H] + calcd. for C 23 H 17 F NO, 342.1289; found: 342.1288.

[0143] Example 5

[0144] According to the reaction conditions of the ring-opening addition of Example 1, different indole heterocyclic compounds were obtained by changing the second substrate, and the specific reaction formula was as follows:

[0145]

[0146] The structural formula of the obtained indole heterocyclic compound and its yield were as follows:

[0147]

[0148] Among them, the structural formula of compound 2-4ak was,

[0149] Chemical name: 2-(2-(bis(4-bromo-1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-one [2-(2-(bis(4-bromo-1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-one] (2-4ak). White solid, 85.5 mg, yield 72%, mp 265.8-266.5 °C; 1H NMR (500 MHz, DMSO-d6) δ 11.17 (s, 1H), 11.17 (s, 1H), 7.47 - 7.40 (m, 3H), 7.34 (d, J = 7.9 Hz, 2H), 7.26 - 7.20 (m, 3H), 7.16 - 7.07 (m, 3H), 7.02 (d, J = 7.3 Hz, 2H), 6.90 (t, J = 7.8 Hz, 2H), 6.78 (d, J = 7.7 Hz, 1H), 6.48 (s, 1H), 6.47 (s, 1H), 4.38 (s, 2H); 13 C NMR (125 MHz, DMSO-d6) δ 197.6, 144.3, 138.3, 136.4, 133.0, 132.7, 130.6, 128.2, 128.1, 127.6, 127.4, 126.3, 125.7, 123.7, 122.7, 122.1, 118.3, 113.1, 111.3, 41.6, 35.7; HRMS (ESI) m / z: [M + Na]+calcd for C31H22Br2N2ONa, 618.9991; found: 618.9991.

[0150] The chemical name of compound 2-4am is,

[0151] The chemical name of compound 2-4am is, 1 H NMR (500 MHz, DMSO-d6) δ 11.17 (s, 1H), 11.17 (s, 1H), 7.47 - 7.40 (m, 3H), 7.34 (d, J = 7.9 Hz, 2H), 7.26 - 7.20 (m, 3H), 7.16 - 7.07 (m, 3H), 7.02 (d, J = 7.3 Hz, 2H), 6.90 (t, J = 7.8 Hz, 2H), 6.78 (d, J = 7.7 Hz, 1H), 6.48 (s, 1H), 6.47 (s, 1H), 4.38 (s, 2H); 13C NMR (125 MHz, DMSO-d6) δ 198.4, 142.6, 136.6, 135.1, 133.8, 133.2, 131.3, 128.6, 128.1, 127.8, 127.5, 126.7, 126.2, 126.1, 123.0, 121.0, 118.1, 116.8, 113.1, 42.3, 35.6; HRMS (ESI) m / z: [M + Na] Calcd for C + C 31 H 22 Cl2N2ONa, 531.1001 ; Found: 531.1002.

[0152] The chemical name of compound 2-4an is,

[0153] The chemical name is: 2-(2-(bis(6-fluoro-1 H-indol-3-yl)methyl)phenyl)-1 - phenylethan-1 -one [2-(2-(Bis(6-fluoro-1 H-indol-3-yl)methyl)phenyl)-1 - phenylethan-1 -one] (2-4an). White solid, 74.9 mg, yield 78%, mp 212.4-213.5 °C; 1 H NMR (500 MHz, DMSO-d6) δ 10.89 (s, 1 H), 10.89 (s, 1 H), 7.90-7.84 (m, 2H), 7.63-7.56 (m, 1 H), 7.48-7.39 (m, 2H), 7.21 -7.19 (m, 1 H), 7.18-7.10 (m, 6H), 7.03 (d, J = 7.8 Hz, 1 H), 6.69 (td, J = 9.8, 2.4 Hz, 2H), 6.59 (s, 1 H), 6.58 (s, 1 H), 5.80 (s, 1 H), 4.40 (s, 2H); 13 C NMR (125 MHz, DMSO-d6) δ 198.4, 158.8 (d, J = 233.9 Hz), 142.8, 136.6, 136.4 (d, J = 12.8 Hz), 133.7, 133.2, 131.2, 128.7, 128.1, 128.0, 126.6, 126.1, 124.9 (d, J = 3.3 Hz), 123.4, 119.9 (d, J = 10.3 Hz), 117.2, 106.7 (d, J = 24.3 Hz), 97.4 (d, J = 25.4 Hz), 42.4, 36.0; 19F NMR (471 MHz, DMSO-d6) δ -122.23 - -122.39 (m, 2F); HRMS (ESI) m / z: [M+Na] calcd for C + calcd. for C 31 H 22 F2N2ONa, 499.1592; found: 499.1592.

[0154] Example 6

[0155] According to the reaction conditions of the ring-opening addition reaction in Example 1, different substituted indole heterocyclic compounds were obtained by changing the first substrate, and the specific reaction formula is as follows:

[0156]

[0157] The structural formula of the obtained polysubstituted indole heterocyclic compound and its yield are as follows:

[0158]

[0159] Preparation of derivatives of compound 2-4aa in Example 7

[0160] Example 7 provides derivatives of the ring-opening addition product 2-4aa. First, 2-4aa (0.50 g, 1.1 mmol) can be reduced by sodium borohydride (NaBH4, 83.2 mg, 2.2 mmol) to reduce the carbonyl group to a hydroxyl group, to obtain compound 2-5 with a yield of 94% (0.47 g); then compound 2-5 (44.2 mg, 0.1 mmol) is reacted with acetic anhydride (Ac2O, 14 μL, 0.15 mmol) in acetonitrile as a solvent, pyridine (Py, 11 μL, 0.12 mmol) as a base, and 4-dimethylaminopyridine (4-DMAP, 1.2 mg, 0.01 mmol) as a catalyst to obtain acylated product 2-6 with a yield of 68% (32.8 mg), and compound 2-7 is reacted with hydroxylamine hydrochloride (10.4 mg, 0.15 mmol) in an ethanol aqueous solution with a yield of 84% (38.5 mg).

[0161] The structural formula of the obtained derivatives is as follows:

[0162]

[0163] Among them, the structural formula of compound 2-5 is,

[0164] Chemical name: 2-(2-(bis(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-ol [2-(2-(Bi(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-ol] (2-5). White solid, yield 94%, mp 112.9-113.4 °C; 1 H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 2H), 7.39-7.31 (m, 3H), 7.29-7.22 (m, 4H), 7.19 (d, J = 7.6 Hz, 3H), 7.11-7.00 (m, 5H), 6.88 (t, J = 7.4 Hz, 2H), 6.61 (s, 1H), 6.58 (s, 1H), 6.18 (s, 1H), 5.41 (d, J = 4.3 Hz, 1H), 4.84-4.76 (m, 1H), 3.01 (dd, J = 13.5, 8.1 Hz, 1H), 2.92 (dd, J = 13.6, 5.0 Hz, 1H); 13 C NMR (125 MHz, DMSO-d6) δ 146.1, 142.9, 136.9, 136.7, 136.7, 130.9, 128.0, 127.9, 126.8, 126.7, 126.6, 125.9, 125.7, 125.4, 124.2, 124.2, 120.9, 119.2, 119.0, 118.2, 118.2, 118.1, 118.0, 111.5, 73.9, 42.8, 35.5; HRMS (ESI) m / z: [M + Na] + calcd. for C 31 H 26 N2ONa, 465.1938; found: 465.1937.

[0165] The structural formula of compound 2-6 is,

[0166] Chemical name: 2-(2-(bis(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-ol [2-(2-(Bi(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-ol] (2-5). White solid, yield 94%, mp 112.9-113.4 °C; 1H NMR (500 MHz, DMSO-d6) δ 10.84 (s, 1H), 10.83 (s, 1H), 7.37 (d, J = 8.1 Hz, 2H), 7.27 (q, J = 6.0 Hz, 4H), 7.20 - 7.14 (m, 4H), 7.13 - 7.07 (m, 1H), 7.09 - 7.02 (m, 4H), 6.92 - 6.83 (m, 2H), 6.63 (s, 1H), 6.58 (s, 1H), 6.10 (s, 1H), 5.90 (t, J = 6.8 Hz, 1H), 3.22 (dd, J = 14.1, 8.2 Hz, 1H), 3.12 (dd, J = 14.1, 5.8 Hz, 1H), 1.95 (s, 3H); 13 C NMR (125 MHz, DMSO-d6) δ 169.5, 142.9, 140.4, 136.7, 136.7, 135.0, 130.6, 128.3, 128.3, 127.9, 126.6, 126.5, 126.3, 126.3, 125.7, 124.3, 124.2, 121.0, 119.0, 119.0, 118.3, 117.7, 117.6, 111.6, 111.5, 76.1, 35.6, 20.9; HRMS (ESI) m / z: [M + Na] + calcd. for C 33 H 28 N2O2Na, 485.2224; found: 485.2226.

[0167] The structural formula of compound 2-7 is,

[0168] Chemical name: 2-(2-(di(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-one oxime [2-(2-(di(1H-indol-3-yl)methyl)phenyl)-1-phenylethan-1-one oxime] (2-7). White solid, yield 84%, mp 109.0-121.1 °C; 1H NMR (500 MHz, DMSO-d6) δ 11.48 (s, 1H), 10.87 (s, 2H), 7.42-7.35 (m, 2H), 7.35-7.31 (m, 2H), 7.31-7.23 (m, 3H), 7.22-7.15 (m, 2H), 7.14-7.08 (m, 1H), 7.08-7.01 (m, 4H), 6.95-6.90 (m, 1H), 6.91-6.84 (m, 2H), 6.77-6.71 (m, 2H), 6.22 (s, 1H), 4.23 (s, 2H); 13 C NMR (125 MHz, DMSO-d6) δ 154.9, 142.6, 136.7, 135.9, 134.3, 128.5, 128.3, 128.2, 127.8, 126.7, 126.0, 125.9, 125.8, 124.3, 121.0, 119.1, 118.3, 117.2, 111.5, 35.8, 27.7; HRMS (ESI) m / z: [M+H] + calcd. for C 31 H 26 N3O, 456.2070; found: 456.2069.

[0169] Example 8

[0170] I. Test cells, animals and reagents

[0171] Experimental cells: MCF-7 human breast cancer cells were purchased from Wuhan University, China Typical Culture Collection Center, and MCF-10A human normal breast epithelial cells were purchased from Sibcro (Shanghai) Biotechnology Co., Ltd.

[0172] Experimental animals: 4-week-old female BALB / c-nu mice were purchased from Hunan Slike Jingda Experimental Animal Co., Ltd. Animal Production License No.: SCXK(Xiang)2019-0004, the animal breeding facility was provided by the Experimental Animal Center of South Central University for Nationalities, and the experimental unit used license number: SYXM(E)2021-0089. This experiment has been approved by the Experimental Animal and Biomedical Ethics Committee of South Central University for Nationalities, approval number: No.2024-SCUEC-067. Before the formal start of the experiment, all mice were adaptively fed in the laboratory environment with a relative humidity of 45% ± 15% and a feeding temperature of 20℃ ± 2℃ for 7 days, with a 12h / 12h light-dark alternation, during which the mice were free to drink water and eat, and the bedding was changed every 2 days.

[0173] Experimental reagents: DMEM medium, PBS solution, trypsin purchased from Gibco; DAPI, BCA protein concentration determination kit purchased from Biyun Tian Biotechnology; Annexin V-FITC / PI double staining cell apoptosis detection kit purchased from Tongren Chemical; Bax, Bcl-2, PARP, Pro caspase 9, P53, Cyclin B1, GAPDH, β-actin purchased from Abotek.

[0174] II. Experimental instruments

[0175] CO2 incubator (Thermo-240i); refrigerated centrifuge (Eppendorf-5425R); multifunctional enzyme marker (SPARK10M, TECAN); confocal microscope (LSM 900, ZEISS); inverted microscope (ECLIPSE Ts2); flow cytometer (FACSCalibur, BD); developing instrument (Omega Lum W).

[0176] III. Cell experiment one

[0177] Compound 2-4ak in Example 5 was taken as the experimental object, and the cytotoxic effect of compound 2-4ak on MCF-7 cells and MCF-10A was determined.

[0178] The experimental method is MTT method

[0179] Logarithmic growth phase cells were trypsinized, uniformly blown and diluted at a certain ratio, 100 μL per well was inoculated into a 96-well plate, and placed in a 37℃, 5% CO2 incubator for 6h, then the old culture medium was discarded, 100 μL of different concentrations of 166 solution (serum-free DMEM medium dilution) was added, incubated for 24h, then MTT solution (0.05mg / mL, serum-free DMEM medium dilution) was added, 4h later, the MTT solution was discarded, 100 μL of DMSO was added to dissolve the crystals, and then the multifunctional enzyme marker was used to detect the absorbance OD value at 570nm after uniform oscillation on the oscillator, and the half maximal inhibitory concentration (IC50) was calculated. IC50=OD drug group / OD blank group*100%.

[0180] See Figure 1 B, C, the experimental results of compound 2-4ak on MCF-7 IC 50 =1.97 μM, and the IC 50 =18.7 μM on MCF-10A.

[0181] According to the same method, the experimental results of the compounds in the derivative examples are recorded as follows Figure 1 A.

[0182] Four, cell experiment two

[0183] The compound 2-4ak in Example 5 was taken as the experimental object, and the effects of the compound 2-4ak on the apoptosis, autophagy and ferroptosis of MCF-7 cells were determined.

[0184] Experimental method:

[0185] (1) Flow cytometry apoptosis detection

[0186] Logarithmic growth phase MCF-7 cells were trypsinized, uniformly blown, diluted at a certain ratio, and inoculated into a six-well plate at 2 mL per well. After 6 h of incubation, the culture medium was discarded, and compound 2-4ak (0, 5, 15, and 30 μM) diluted in serum-free medium was added to the experimental group, and the blank group was added with serum-free DMEM medium. After 24 h of treatment, the cells were collected by trypsinization, centrifuged at 1000 rpm at room temperature for 5 min, resuspended with pre-cooled PBS, centrifuged again at 1000 rpm for 5 min, and then 100 μL of Binding Buffer was added to suspend the cells. After adding 5 μL of Annexin V-FITC to the working solution and mixing well, 5 μL of PI staining solution was added and mixed well, and then incubated at room temperature for 15 min in the dark. After incubation, 400 μL of Binding buffer working solution was added, and the sample was detected by flow cytometry.

[0187] (2) DAPI staining method

[0188] Logarithmic growth phase MCF-7 cells were selected, inoculated in a laser confocal culture dish, and cultured in a constant temperature incubator. When the cell confluence reached about 80%, the old culture medium was discarded, and 2-4ak solution (0 μM, 5 μM, 10 μM, and 20 μM) prepared with serum-free DMEM medium was added. After 24 h of incubation, the adherent cells were gently washed with 4°C pre-cooled PBS solution for 2 times, and 1 mL of 4% paraformaldehyde was added for fixation for 15 min. After fixation, 1 mL of DAPI staining solution was added for staining at room temperature for 5 min in the dark, and then washed with PBS solution for 3 times. Laser confocal microscopy was used for observation.

[0189] (3) Transmission electron microscopy analysis

[0190] MCF-7 cells were evenly inoculated in 10 cm culture dishes, and when the cell confluence reached 50%, 2-4ak solution (0 μM and 4 μM) prepared in serum-free DMEM was added for 24 h. After trypsin digestion and blowing, the cells were centrifuged, the supernatant was discarded, and the cells were resuspended in electron microscopy fixing solution and fixed at 4°C for 4 h. After fixation, the fixing solution was removed by centrifugation, 0.1 M phosphate buffer was added, and the cells were resuspended and centrifuged. The washing was repeated 3 times. 1% agarose solution was added to the EP tube and dissolved in advance, and the cells were wrapped in agarose. Then, 1% osmium acid was used for room temperature dark fixation for 2 h. PB solution was used for repeated washing 3 times, 15 min each time. Dehydration was performed at room temperature, and then embedding agent was added for embedding. The cells were sectioned using an ultrathin microtome, and 2% uranyl acetate saturated alcohol solution was used for room temperature dark staining for 8 min. The cells were sequentially washed with 70% alcohol and ultrapure water 3 times, and then 2.6% citric acid lead solution was added for air exclusion staining for 8 min. The cells were washed with ultrapure water 3 times, and then dried at room temperature. Finally, the cells were observed under a transmission electron microscope and photographed for analysis.

[0191] (4) Western blotting

[0192] MCF-7 cells were evenly inoculated in 10 cm culture dishes, and when the cell confluence reached 50%, 2-4ak solution (0 μM, 5 μM, 10 μM and 15 μM) prepared in serum-free DMEM was added for 24 h. The culture medium was collected into the corresponding EP tube, and 2 mL of trypsin was added to the culture dish to digest the cells. After most of the cells were rounded off and fell off, 4 mL of DMEM complete medium was added to terminate the digestion, and then the cells were transferred to the EP tube in the previous step, centrifuged at 1000 rpm for 5 min to collect the cell precipitate. The cell precipitate was resuspended and washed with 1 mL of PBS solution, repeated 2 times, and then 500 μL of RIPA lysis buffer containing protease inhibitors was added. The cells were lysed on ice for 30 min, and during this period, the cells were shaken on a shaker for 10 s every 5 min. After complete lysis, the cells were centrifuged at 4°C, 12000 rpm for 15 min, and the supernatant was collected and the volume was recorded. Protein quantification was performed according to the BCA protein concentration determination kit instructions. The separation gel and the concentrated gel were prepared, and then an appropriate amount of protein was added to each well for electrophoresis and membrane transfer. After blocking with 10 mL of fast blocking solution, the primary antibody was added for incubation overnight, and then the cells were washed with TBST for 3 times. The corresponding secondary antibody was added for incubation at room temperature for 1 h. After washing again, the developing solution was added, and the freshly prepared ECL luminescent solution was evenly covered on the front surface of the PVDF membrane. The developer was used for exposure observation.

[0193] Five, animal experiment

[0194] The compound 2-4ak in Example 5 was used as the experimental object, and a MCF-7 transplanted tumor model was established.

[0195] Experimental method:

[0196] (1) Construction of MCF-7 tumor xenograft model in nude mice

[0197] The well-grown MCF-7 cells were trypsinized and terminated by adding DMEM complete medium, centrifuged at 1000 rpm for 3 min to remove trypsin, and resuspended in PBS to prepare a cell suspension with a concentration of 2 x 10 7 A 120 μL aliquot of the MCF-7 cell suspension was injected subcutaneously into the right back of the mouse to establish the MCF-7 tumor xenograft model.

[0198] (2) Grouping and scheme of animal experiment

[0199] The growth of the subcutaneous tumor in the mice inoculated with the MCF-7 cell suspension was closely observed, and the mice were randomly divided into the PBS group and the drug administration group (2-4ak, 25 mg / kg) when the tumor volume reached 50 mm 3 After 14 days, the experiment was terminated, the mice were sacrificed under anesthesia, the tumor tissues were peeled off and weighed, photographed, and the main organs (heart, liver, spleen, lung, kidney) of the mice were taken together with the tumor tissues for H&E staining. The tumor volume was calculated according to the following formula: tumor volume V = (L x W 2 ) / 2, wherein L (mm) is the longest diameter of the tumor and W (mm) is perpendicular to L (mm).

[0200] (3) H&E, TUNEL and immunofluorescence staining of tissues

[0201] H&E staining: The collected tumor tissues and main organs of the mice were fixed with 4% paraformaldehyde, dehydrated and embedded in wax, embedded in an embedding machine, cut into 4 μm thick sections with a paraffin sectioning machine. The sections were placed in a 40°C water bath for expansion, the tissue sections were taken out with a glass slide to evenly spread them on the glass slide, and finally sent to a 60°C oven for baking. After deparaffinization, the sections were sequentially placed in hematoxylin staining solution for 5 min, dehydrated with 95% alcohol for 1 min, and finally placed in eosin staining solution for 15 s. After dehydration and mounting, they were observed under a microscope.

[0202] TUNEL staining: The paraffin sections were prepared and deparaffinized as described above. After deparaffinization, proteinase K solution was added and incubated for 20 min, then washed with PBS, and 0.1% Triton solution was added and incubated for 20 min to break the membrane. TUNEL staining was performed according to the instructions of the TUNEL kit. After staining, the sections were washed with PBS for 3 times, DAPI staining solution was added for counterstaining of the cell nuclei, and anti-fluorescence quenching mounting medium was used for mounting. The sections were observed under a fluorescence microscope.

[0203] Immunofluorescence staining: The paraffin section making and dewaxing process were the same as above. After the paraffin section was dewaxed, citric acid antigen repair solution was added for repair. After repair, PBS solution was washed 3 times, each for 5 min, and then BSA was blocked for 30 min. After blocking, different sources of Ki-67 and CD31 primary antibody mixed solution were added and incubated at 4°C overnight. The next day, PBS solution was washed 3 times, and the corresponding secondary antibody was added and incubated at room temperature for 1 h, and then PBS was added for washing, and DAPI was added for incubation at room temperature for 10 min for cell nucleus restaining. After all staining was completed, PBS solution was washed 3 times, spontaneous fluorescence quencher was added and incubated for 10 min, and anti-fluorescence quenching mounting agent was used for mounting, and then observed under a fluorescence microscope.

[0204] Six, experimental results

[0205] Cell experiment two:

[0206] (1) Flow cytometry cell apoptosis detection result analysis

[0207] The apoptosis of MCF-7 cells treated by 2-4ak was detected by flow cytometry to verify whether 2-4ak could inhibit cell proliferation by inducing apoptosis of MCF-7 cells. As shown in Figure 2 , after treatment with 2, 4 and 8 μM of 2-4ak, the relative proportion of apoptotic cells in MCF-7 cells showed an increasing trend, and had a concentration-dependent effect, and the total percentage of early and late apoptotic cells increased from 8.84% to 9.63, 15.05 and 24.8%, indicating that 2-4ak could induce apoptosis of MCF-7 cells.

[0208] (2) DAPI staining method result analysis

[0209] After MCF-7 cells treated with different concentrations of 2-4ak (0, 2, 4 and 8 μM) were incubated for 24 h, the changes in nuclear morphology were observed under a laser confocal microscope by DAPI staining. As shown in Figure 3 , the nuclei of untreated MCF-7 cells were regular in shape and clear in outline, while the MCF-7 cells treated with 2-4ak showed nuclear condensation and highly condensed chromatin, as indicated by the red arrow, which was a characteristic of early apoptosis. With the increase of 2-4ak concentration, the nuclei of some MCF-7 cells were fragmented at 8 μM, forming round bodies of different sizes, as indicated by the white arrow, which was a characteristic of late apoptosis.

[0210] (3) Transmission electron microscopy result analysis

[0211] To verify the effect of 2-4ak on autophagy of human breast cancer cells MCF-7, first observe the effect of 2-4ak on the ultrastructure of MCF-7 cells by transmission electron microscopy. MCF-7 cells were treated with different concentrations of 2-4ak (0 and 4 μM), and after 24 h of action, the cells were collected, fixed with electron microscope fixing solution, and the ultrastructure of MCF-7 cells was observed by transmission electron microscopy, and the results are shown in Figure 4 Compared with normal MCF-7 cells, most of the mitochondria treated with 2-4ak were reduced, the mitochondrial cristae were reduced (yellow arrow), the nuclear chromatin was uniform, and there was no chromatin condensation, which was a characteristic of ferroptosis of the cells. At the same time, four autophagolysosome structures (red arrow) were observed in the field of view of the cells treated with 2-4ak, while no typical autophagy structure was observed in the normal cells. The above results show that 2-4ak can induce MCF-7 cells to undergo cell ferroptosis and autophagy.

[0212] (4) Western blotting result analysis

[0213] Caspase 8 is an apoptosis initiator in the caspase family of proteins. After activation, caspase 8 is cleaved to transmit apoptosis signals to the downstream, leading to the activation of apoptosis effectors caspase 3 or caspase 7, causing exogenous apoptosis and mediating the cleavage of downstream proteins such as PARP. As shown in Figure 5 Compared with the Control group, the expression of caspase 8 protein in MCF-7 cells treated with 2-4ak increased slightly and then decreased significantly with increasing concentration, while PARP showed the same trend and decreased significantly in the highest concentration group, indicating that 2-4ak may activate exogenous apoptosis by activating caspase 8, thereby inhibiting the growth of MCF-7 cells.

[0214] In autophagy, p62 is selectively wrapped into autophagosomes as a bridge between LC3 and polyubiquitinated proteins, and is degraded by autolysosomal proteolytic enzymes, and its expression level is negatively correlated with the intensity of autophagy. Beclin 1 is an important autophagy-related protein that participates in autophagosome formation and is essential for autophagy. LC3 I is modified to LC3 II by the ubiquitin-like system, and the change of LC3 II / I ratio is required to judge autophagy, which is positively correlated with the level of autophagy and is a representative indicator for detecting autophagy. GPX4 is the core of ferroptosis regulation, and its inactivation leads to the accumulation of lipid peroxides in cells, triggering ferroptosis, which is a key indicator for judging ferroptosis. As shown in Figure 5B shows that compared with the Control group, the expression level of autophagy substrate p62 in MCF-7 cells treated by 2-4ak decreased significantly in a dose-dependent manner, while the total Beclin 1 showed an upward trend, and the ratio of autophagy marker LC3 II / I increased significantly, proving that 2-4ak treatment can induce autophagy in MCF-7 cells, achieving the effect of inhibiting the proliferation of MCF-7 cells. The expression of GPX4 also showed a downward trend with the increase of 2-4ak treatment concentration, confirming that ferroptosis also plays a role in the process of 2-4ak inhibiting the proliferation of MCF-7 cells.

[0215] Animal experiment:

[0216] (1) MCF-7 tumor model results analysis

[0217] After subcutaneously injecting MCF-7 cells into the right back of mice for 7 days, the tumor volume of mice reached 50 mm3, and the mice were randomly divided into 2 groups: PBS group and 2-4ak 25 mg / kg group (n=5). The tumor diameters of mice in each group were measured every 2 days with vernier caliper to calculate the tumor volume of mice, and the tumor volume-time curve of mice was drawn, as shown in Figure 6 A. At the end of the experiment, the tumor tissues of mice in each group were taken and photographed, as shown in Figure 6 B. Compared with the PBS group, the growth of tumor tissues in the 2-4ak treatment group was significantly inhibited. Figure 6 B also confirmed that the tumor volume of mice in the 2-4ak treatment group was significantly lower than that in the PBS group at the end of the experiment, showing significant anti-breast cancer activity.

[0218] (2) H&E, TUNEL and immunofluorescence staining of tumor tissues of MCF-7 tumor-bearing mice

[0219] To further detect the anti-breast cancer effect of 2-4ak in mice, we performed H&E staining on the tumor tissues of each group to observe the tissue morphology Figure 7 A). From the figure, it can be seen that the tumor cell structure in the PBS group is orderly and densely arranged, with large and deeply stained nuclei and less cytoplasm. While in the 2-4ak treatment group, the tumor cell structure is loose, the cell nucleus is obviously dissolved, and the chromatin is condensed, with a large number of necrotic cells. The above results show that 2-4ak can effectively inhibit the proliferation of breast cancer cells.

[0220] TUNEL staining is the most commonly used method for detecting apoptosis in tissue samples. Its principle is to use fluorescently labeled deoxyuridine triphosphate to connect to the 3'-OH end of the broken DNA of apoptotic cells, and to detect apoptotic cells by fluorescence excitation. The higher the TUNEL positive cell rate, the higher the proportion of apoptotic cells. TUNEL staining has high sensitivity and strong specificity, and can accurately reflect the most typical biochemical and morphological characteristics of cell apoptosis. For example,Figure 7 As shown in FIG. 2-4A, the number of apoptotic positive cells (green fluorescence) in the tumor tissue sections of the 2-4ak treatment group mice was significantly increased relative to the PBS group, indicating that 2-4ak can effectively induce cancer cell apoptosis to exert its anticancer effect.

[0221] Ki-67 is a marker of cell proliferation, present in all active stages of the cell cycle, and is not expressed in the G0 phase. Therefore, the expression level of Ki-67 can represent the degree of cell proliferation activity, and a higher expression level of Ki-67 indicates more cells in the division phase and more active cell proliferation. CD31 is a platelet-endothelial cell adhesion molecule, which marks the differentiation of vascular endothelial cells and is used to assess the generation of blood vessels in tumors. A higher positive expression indicates faster tumor proliferation. As shown in FIG. 2-4B, the positive expression of Ki-67 (red) and CD31 (green) in the PBS group was significantly higher than that in the 2-4ak group, indicating that 2-4ak can effectively inhibit the proliferation of tumor cells to delay the growth of mouse tumor tissue. Figure 7

[0222] (3) Biological safety evaluation

[0223] To evaluate the biological safety of 2-4ak, the body weight and major organs of the mice in each group were evaluated. H&E staining of the heart, liver, spleen, lung, and kidney tissues of the mice showed no obvious pathological changes. Figure 8

[0224] In summary, the synthesis method of the two polyaryl-substituted indole heterocyclic compounds provided in the present application is as follows: the first substrate isobenzopyran and the second substrate indole reacts under the conditions of acetonitrile and racemic binol phosphate to generate 3-(3-phenyl-1H-isochromen-1-yl)-1H-indole reacts under the conditions of acetonitrile and aluminum chloride to generate 2-(2-(di(1H-indol-3-yl)methyl)phenyl)-1-phenyl ethanone This method develops a selective Friedel-Crafts reaction of isobenzopyran and indole, and obtains a series of addition and Friedel-Crafts ring-opening products in moderate to good yields, which proves the good tolerance of each substrate in the reaction. The gram-scale reaction and derivatization experiments also prove that the series of compounds have good applicability. By combining the two structures of indole and isobenzopyran, which have good biological activity, new compounds with cytotoxic activity to the MCF-7 cell line are successfully obtained.

[0225] ​​The above merely provides the preferred embodiments of the present application, and is not used to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modifications, equivalent replacements, improvements, etc. made within the principles and technical scope of the present application shall fall into the scope of the present application.

Claims

1. A compound represented by Formula (I) and / or Formula (II) or a pharmaceutically acceptable salt thereof: Formula (I) Formula (II) wherein R is selected from the group consisting of no substitution or optionally substituted by one, two, or more R 2 substituted C 1-10 alkyl, C 1-10 alkoxy, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl; Each R 1 They may be the same or different, and are independently selected from H, halogens, OH, CN, NO2, unsubstituted, or optionally substituted by one, two, or more Rs. a The following groups are substituted: C 1-10 Alkyl, C 1-10 Alkoxy, C 3-10 cycloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclic; each R is independently selected from the group consisting of H, halogen, OH, CN, N02, unsubstituted or optionally substituted with one, two, or more R 2 the same or different, are independently selected from the group consisting of H, halogen, OH, CN, N02, unsubstituted or optionally substituted with one, two, or more R b substituted with one, two, or more R 1-10 alkyl, C 1-10 alkoxy, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl; each R is independently selected from the group consisting of H, halogen, CN, N02, unsubstituted or optionally substituted with one, two, or more R 3 the same or different, are independently selected from the group consisting of H, halogen, CN, N02, unsubstituted or optionally substituted with one, two, or more R c substituted with one, two, or more R 1-10 alkyl, C 1-10 alkoxy, C 3-10 cycloalkyl, C 6-10 aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclyl, NH2; Each R 4 They may be identical or different, independently selected from H, without substitution, or optionally by one, two, or more Rs. d The following groups are substituted: C 1-10 Alkyl, C 3-10 cycloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclic; Each R a R b R c R d They may be the same or different, and are independently selected from H, halogens, OH, CN, NO2, and C. 1-10 Alkyl, C 1-10 Alkoxy, C 3-10 cycloalkyl, C 6-10 Aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclic, C 1-6 Alkyl-C 6-10 Aryl-S(O)2-; m is selected from 0, 1, 2, 3, 4 or 5; n is selected from 0, 1, 2, 3, 4 or 5; p is selected from 0, 1, 2, 3, 4 or 5.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, R is selected from the group consisting of no substitution or optionally substituted by one, two, or more R 2 substituted by one, two, or more R 1-6 alkyl, phenyl; Preferably, R is selected from C 1-6 alkyl, Preferably, R is selected from n-butyl, t-butyl, Preferably, each R 1 identically or differently, independently of one another, selected from H, halogen, C 1-6 alkyl, C 1-6 alkoxy; Preferably, each R 1 are the same or different, independently of each other, selected from H, F, methoxy; Preferably, each R 2 They are either the same or different, and are independently selected from H, halogens, and C. 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, phenyl; Preferably, each R 2 are the same or different, independently of each other, selected from H, F, CI, methoxy, trifluoromethyl, phenyl; Preferably, each R 3 They may be the same or different, and are independently selected from H, halogens, OH, CN, and C. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkyl-phenyl-S(O)2-NH-, phenyl-C 1-6 Alkoxy; Preferably, each R 3 are the same or different, independently of one another, selected from the group consisting of H, F, CI, Br, OH, CN, methyl, methoxy, 4-methylphenylsulfonyl-NH-, benzyloxy; Preferably, each R 4 are the same or different, independently of each other, selected from H, methyl; Preferably, each R a , R b , R c , R d are the same or different, independently of each other, selected from H, halogen, OH, CN, N02, C 1-6 alkyl, C 1-6 alkoxy, phenyl, 5-6 membered heteroaryl, 3-6 membered heterocyclyl, C 1-6 alkyl-phenyl-S(0)2-; Preferably, each R a , R b , R c , R d are the same or different, independently of each other, selected from the group consisting of H, halogen, OH, CN, NO2, methyl, methoxy, phenyl, 4-methylphenylsulfonyl.

3. The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, characterized in that, The compound represented by Formula (I) and / or Formula (II) is selected from the following structures: wherein R 1 , R 2 , R 3 , R 4 each independently have the definition of claim 1 or 2.

4. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein The compound represented by Formula (I) is selected from the following structures: The compound represented by Formula (II) is selected from the following structures:

5. A process for the preparation of a compound of formula (I) and / or formula (II) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 4, comprising the steps of: reacting a first substrate compound a and a second substrate compound b in the presence of a catalyst to obtain a compound represented by Formula (I) and / or Formula (II); wherein R, R 1 , R 3 , R 4 , m, p independently of one another have the definitions according to any of claims 1 to 4; L is selected from the group consisting of C 1-6 alkoxy, for example methoxy; Preferably, the catalyst is selected from catalyst 1 or catalyst 2; wherein catalyst 1 is selected from rac-BPA TsOH, CF3CO2H, CF3SO3H, Cu(OTf)2; catalyst 2 is selected from AlCl3; when the catalyst is catalyst 1, the reaction produces compounds of formula (I) and (II); when the catalyst is catalyst 2, the reaction produces compounds of formula (II); Preferably, the molar ratio of compound a, compound b and catalyst is selected from (0.2-5):1:(0.01-0.1), for example (0.9-3):1:(0.02-0.0.05), (0.2-0.8):1:(0.01-0.1), for example 1.5:1:0.025, 0.5:1:0.025; Preferably, the molar ratio of compound a, compound b and catalyst 1 is selected from (0.9-3):1:(0.02-0.0.05), for example 1.5:1:0.025; Preferably, the molar ratio of compound a, compound b and catalyst 2 is selected from (0.2-0.8):1:(0.01-0.1), for example 0.5:1:0.025; Preferably, the reaction is carried out in the presence of an organic solvent selected from at least one of DCM, toluene, methanol, ethanol, THF, 2-methyltetrahydrofuran, 1,4-dioxane, acetonitrile, DMF.

6. Use of a compound represented by Formula (II) of any one of claims 1-4 as a starting material for further preparation of a derivative thereof.

7. A method for further preparation of a derivative of a compound represented by Formula (II) of any one of claims 1-4 using a compound represented by Formula (II) as a starting material, comprising the following Scheme I, Scheme II or Scheme III: Scheme I: reacting a compound represented by Formula (II) in the presence of a reducing agent to obtain a compound represented by Formula (II-a); wherein R, R 1 R, R 3 R, R 4 m, p independently of one another have the definitions according to any of claims 1 to 4; Preferably, the reducing agent is selected from at least one of sodium borohydride, potassium borohydride, sodium borohydride acetate, sodium cyanoborohydride; Scheme II: reacting a compound represented by Formula (II-a) with acetic anhydride to obtain a compound represented by Formula (II-b); wherein R, R 1 , R 3 , R 4 , m, p have independently of each other the definitions according to any of claims 1 to 4; Scheme III: reacting a compound represented by Formula (II) with hydroxylamine hydrochloride to obtain a compound represented by Formula (II-c); wherein R, R 1 , R 3 , R 4 , m, p have independently of each other the definitions according to any of claims 1 to 4.

8. A compound represented by Formula (II-a) or Formula (II-b) or Formula (II-c): wherein R, R 1 , R 3 , R 4 , m, p independently of one another have the definitions given in any of claims 1 to 4.

9. A pharmaceutical composition comprising at least one of the compound represented by Formula (I) and / or Formula (II) or a pharmaceutically acceptable salt thereof.

10. Use of at least one of the compound represented by Formula (I) and / or Formula (II) or a pharmaceutically acceptable salt thereof of any one of claims 1-4 in the preparation of a medicament for the prevention and / or treatment of cancer; Preferably, the cancer is breast cancer.

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