Synthesis and application of diheterocyclic molecular compound containing pyrroline and pyridine

Abstract

The invention belongs to the technical field of medicinal chemistry, and particularly relates to synthesis and application of a diheterocyclic molecular compound containing pyrroline and pyridine. The diheterocyclic molecular compound containing pyrroline and pyridine and pharmaceutically acceptable salts thereof are synthesized for the first time, and the compound is found to have good anti-tumor activity and can inhibit proliferation and migration of various tumor cells. Therefore, the compound synthesized by the invention has a good application prospect in the aspect of new anti-tumor drugs, so that a foundation is laid for development of corresponding anti-cancer drugs, particularly colorectal cancer drugs, and meanwhile, the compound has important significance in structural modification of pyridine-containing drug molecules.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicinal chemistry, and in particular relates to the synthesis and application of a bicyclic heterocyclic molecular compound containing dihydropyrrole and pyridine. Background Art

[0002] Pyridine and its derivatives are the most common nitrogen heterocyclic structures in drug molecules, accounting for more than 80% of the small molecule drugs approved in recent years. This type of compound has outstanding performance in the fields of anti-tumor and antiviral. However, the regioselective modification of the pyridine ring, especially the C4 alkylation reaction, has always faced the dual challenges of selectivity control and functional group compatibility. The traditional Minisci reaction requires strongly acidic conditions and is prone to produce a mixture of positional isomers. The N-functionalized pyridinium salt strategy developed in recent years has provided a new approach to this problem. γ,δ-unsaturated α-imidooxo acids, as novel free radical precursors, undergo single-electron oxidative decarboxylation under photocatalytic conditions. The resulting imine radical undergoes 5-exo-trig cyclization to form a dihydropyrrolidine skeleton with important pharmaceutical activity. This type of structure is widely found in various clinical drugs, such as the antiviral drug nirmatrelvir.

[0003] This application has developed a photocatalytic tandem reaction strategy by combining the above two important reaction systems. The reaction uses γ,δ-unsaturated α-imidooxo acid and N-aminopyridinium salt as substrates. Through the visible light-induced single electron transfer process, the generation and cyclization of imine radicals are first realized, and then the formed dihydropyrrolidino radical is regioselectively added to the C4 position of the pyridinium salt. This process not only solves the selectivity problem of the traditional Minisci reaction, but also realizes the simultaneous construction of pyridine ring and pyrrolidine structure. The reaction does not require transition metal catalysis, the conditions are mild, and the functional group compatibility is good, providing an efficient and green new route for the synthesis of complex nitrogen heterocyclic structures. This one-step strategy for constructing dinitrogen heterocycles is particularly suitable for the later modification of drug molecules.

[0004] Preliminary biological activity showed that the key compound had a certain inhibitory effect on colorectal cancer cells HCT-116. This strategy uses light energy to drive the cyclization and pyridation reactions of imine radicals, providing an important synthetic tool for innovative drug development. Summary of the Invention

[0005] The purpose of the present invention is to provide a synthesis and application of a bicyclic heterocyclic molecular compound containing dihydropyrrole and pyridine, so as to achieve an effective anti-tumor effect.

[0006] To this end, the present invention provides the following technical solutions.

[0007] The first aspect of the present invention provides a compound as shown in Formula III or a pharmaceutically acceptable salt thereof: Among them, R 1 Refers to any of alkyl and aryl groups; R 2 Refers to either fluorine or hydrogen; R 3 、R 4 Each refers to any of aryl, methoxy, methyl, and hydrogen.

[0008] A second aspect of the present invention provides a method for preparing a compound represented by Formula III or a pharmaceutically acceptable salt thereof, the method comprising the following steps: S1: γ,δ-unsaturated α-imidooxo acid, N-aminopyridinium salt, sodium acetate and Mes-Acr + ClO4 - mixing to obtain a mixed solution; S2: Under an inert atmosphere, adding an organic solvent to the obtained mixed solution and stirring the mixture under blue light irradiation to obtain a reaction solution; S3: The obtained reaction solution is concentrated and purified to obtain the compound represented by formula III.

[0009] In some embodiments of the present invention, in step S2, the inert atmosphere is selected from any one of nitrogen and argon.

[0010] In some embodiments of the present invention, in step S2, the organic solvent is selected from any one of acetonitrile, 1,2-dichloroethane, and dichloromethane.

[0011] Preferably, the organic solvent is acetonitrile.

[0012] In some embodiments of the present invention, in step S2, the blue light irradiation conditions are: wavelength 424-525nm, light intensity 10-40W.

[0013] Preferably, the blue light irradiation conditions are: wavelength 424nm, 440nm, 456nm, 525nm, preferably 456nm, and light intensity 40W.

[0014] In some embodiments of the present invention, in step S2, the stirring reaction conditions are: room temperature, time 8-24 hours.

[0015] Preferably, the stirring reaction conditions are: room temperature, time 16 hours.

[0016] The third aspect of the present invention provides a pharmaceutical composition comprising: (1) the compound of formula III as described above or a pharmaceutically acceptable salt thereof; and (2) Pharmaceutically acceptable carriers and / or excipients.

[0017] Another aspect of the present invention provides a use of the compound as described above as shown in Formula III or a pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a drug for treating and / or preventing diseases associated with abnormal cell activity.

[0018] In some embodiments of the present invention, the disease associated with abnormal cell activity is selected from tumors.

[0019] In some embodiments of the invention, the tumor is colorectal cancer.

[0020] By means of the above technical solution, the present invention has at least the following advantages: The present invention introduces a redox-active carboxyl leaving group into the imine structure and utilizes a single electron transfer process mediated by photoredox catalysis to promote the cleavage of the N-LG bond. This process first releases CO2 through a decarboxylation reaction, which then undergoes two β-scissions to efficiently generate an imine radical. The resulting radical undergoes intramolecular cyclization to form a tertiary carbon-centered radical containing a dihydropyrrole skeleton. This active intermediate can regioselectively attack the C-4 position of the N-aminopyridinium salt, ultimately constructing a compound with a bicyclic structure containing both pyrrole and pyridine rings. The preparation method of the present invention is characterized by mild conditions, high atom economy, and simple steps, providing a new approach for the efficient synthesis of complex nitrogen polyheterocycles.

[0021] The compounds synthesized in the present invention, which have a bicyclic structure of pyrrole and pyridine rings, and their pharmaceutically acceptable salts, exhibit excellent antitumor activity and are capable of inhibiting the proliferation and migration of various tumor cells. Therefore, the compounds synthesized in the present invention have promising application prospects in new antitumor drugs, laying the foundation for the development of corresponding anticancer drugs, particularly colorectal cancer drugs. They also have important implications for the structural modification of pyridine-containing drug molecules.

[0022] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention. DETAILED DESCRIPTION

[0023] In order to make the technical means, creative features, objectives and effects achieved by the present invention easier to understand, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0024] Explanation of terms: Unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the laboratory procedures for cell culture, molecular genetics, nucleic acid chemistry, and immunology used herein are conventional procedures widely used in the relevant fields. To facilitate a better understanding of the present invention, definitions and explanations of relevant terms are provided below.

[0025] The term "pharmaceutically acceptable" means relatively non-toxic, safe, and suitable for use by patients.

[0026] The term "pharmaceutically acceptable salt" refers to a salt formed by reacting a compound with a pharmaceutically acceptable acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition).

[0027] The term "pharmaceutically acceptable" means that the carrier or excipient is compatible with the other ingredients of the composition and not largely toxic to the recipient thereof, and / or such carrier or excipient is approved or acceptable for inclusion in pharmaceutical compositions for parenteral administration to humans.

[0028] In some embodiments, carriers or excipients used with the compositions disclosed herein include, but are not limited to, sterile liquids such as water and oils, including oils of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. In some embodiments, when the pharmaceutical composition is administered intravenously, the carrier may be water. Saline solutions, aqueous dextrose solutions, and glycerol solutions may also be used as liquid carriers, particularly for injectable solutions. Examples of suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences by E. W. Martin, which is incorporated herein by reference. Such compositions will contain a clinically effective dose of an antibody or antibody fragment, together with a suitable carrier, to provide a dosage form suitable for the patient. The formulation should be suitable for the intended mode of administration. The formulation may be packaged in ampoules, disposable syringes, or multidose vials made of glass or plastic. For details, see the Pharmacopoeia of the People's Republic of China (2020 Edition) and the Handbook of Pharmaceutical Excipients (Paul J Sheskey, Bruno C Hancock, Gary P Moss, David J Goldfarb, 2020, 9th Edition).

[0029] The term "treat" refers to eliminating the cause or alleviating the symptoms of a disease.

[0030] The term "prevent" refers to reducing the risk of developing a disease.

[0031] The term "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0032] The methods of the present invention are described below by way of specific examples. It should be understood that these examples are intended to illustrate the basic principles, main features, and advantages of the present invention, and the present invention is not limited in scope by the following examples. The implementation conditions used in the examples can be further adjusted according to specific requirements. Unspecified implementation conditions are generally those used in routine experiments or selected according to product specifications. The reagents and raw materials used in the present invention are all commercially available.

[0033] The following embodiments relate to and mention: 1. Raw materials (1) Synthesis of γ, δ-unsaturated α-iminooxacids: In a round-bottom flask, a γ,δ-unsaturated ketone (1.0 equiv.) and methanol were stirred and dissolved at room temperature. α-Aminooxacid hydrochloride (1.0 equiv.) and sodium acetate (2.4 equiv.) were then added. The mixture was moved to a 90°C oil bath, refluxed, and stirred overnight. The reaction was monitored by thin-layer chromatography using dichloromethane / methanol (20:1). After stirring, the reaction mixture was cooled to room temperature and the methanol was removed by rotary evaporation in vacuo to obtain the crude product, which was then purified by silica gel column chromatography (40:1 v / v dichloromethane / methanol) to afford the γ,δ-unsaturated α-imidooxacid.

[0034] (2) Synthesis of N-toluenesulfonyl-1-aminopyridine ylide: Dissolve N-aminopyridine salt (1.0 equiv.) in dichloromethane, stir at room temperature, and cool to 0°C. Slowly add triethylamine (2.2 equiv.) dropwise. Stir for 1-2 minutes, then add p-toluenesulfonyl chloride (1.1 equiv.) and stir overnight at room temperature. After the reaction, extract the mixture three times with dichloromethane and water, then wash once with saturated sodium chloride solution. The combined organic layers are dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting mixture is purified by silica gel column chromatography (20:1 v / v dichloromethane / methanol) to afford N-p-toluenesulfonyl-1-aminopyridine ylide.

[0035] (3) Synthesis of N-aminopyridinium salt: Dissolve N-toluenesulfonyl-1-aminopyridine ylide (1.0 equiv.) in dichloromethane with stirring. Stir for a while, then add trimethyloxonium tetrafluoroborate (1.0 equiv.). Stir overnight at room temperature under nitrogen. Concentrate by rotary evaporation to obtain the crude product, which is then recrystallized to obtain the N-aminopyridinium salt.

[0036] (4) 10-Methyl-9-mesityl acridine perchlorate: Product No. W5100612500, purchased from Anaiji Chemical.

[0037] 2. Instruments (1) Microplate reader: Tecan infinite M1000 Pro.

[0038] (2) The nuclear magnetic resonance data were detected using a German Bruker magnetic resonance spectrometer 500 MHz (AVANCE NEO Ascend 500) or 600 MHz (BrukerAVANCE III HD, equipped with an ultra-low temperature probe).

[0039] (3) Mass spectrometry was performed using an AB Sciex high-resolution mass spectrometer (AB SCIEX 500R).

[0040] Example 1: Preparation of Bis-Heterocyclic Compound 3a In a 10 mL reaction tube, γ, δ-unsaturated α-imidooxacid 1a (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and 10-methyl-9-mesityl acridine perchlorate (Mes-Acr + ClO4 - ) (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the obtained mixed solution, and then magnetic stirring was carried out at room temperature for 16 h under blue light (wavelength 456 nm, light intensity 40 W). After the reaction, the obtained reaction solution was extracted with ethyl acetate (3 × 25 mL), and the combined organic phase was washed with saturated brine (1 × 30 mL), then dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 44.9 mg of the bicyclic heterocyclic compound 3a as a white solid with a yield of 85%.

[0041] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3a are: 1 H NMR (500 MHz, Chloroform- d ) δ 8.48 (d, J = 5.7 Hz, 2H), 7.81 (dd, J = 7.9, 1.8 Hz, 2H), 7.44 – 7.36 (m, 2H), 7.33 (d, J = 6.4 Hz, 3H), 4.38(tt, J = 7.6, 2.2 Hz, 1H), 2.85 – 2.73 (m, 1H), 2.71 – 2.61 (m, 1H), 1.95 –1.87 (m, 1H), 1.49 (s, 3H), 1.48 – 1.43 (m, 1H), 1.41 (s, 3H). 13C{1H} NMR (126 MHz, Chloroform- d) δ 173.05, 156.57, 149.21, 134.48,130.39, 128.37, 127.58, 122.40, 81.77, 42.16, 35.14, 26.29, 24.57, 24.11. HRMS (ESI) Calcd for C 18 H 20 N2: [M+H] + = 265.1699 . Found: 265.1709. Example 2: Preparation of Bis-Heterocyclic Compound 3b In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1b (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 50.7 mg of the bicyclic heterocyclic compound 3b as a yellow solid with a yield of 85%.

[0042] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3b are: 1 H NMR (600 MHz, Chloroform- d ) δ 8.47 (d, J = 4.2 Hz, 2H), 7.73 (d, J = 8.5 Hz, 2H), 7.35 (d, J = 8.5 Hz, 2H), 7.30 (d, J = 5.4 Hz, 2H), 4.36 (t, J= 7.7 Hz, 1H), 2.78 – 2.69 (m, 1H), 2.65 – 2.57 (m, 1H), 1.94 – 1.86 (m, 1H), 1.48-1.43 (m, 4H), 1.39 (s, 3H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 171.88, 156.43, 149.07, 136.40,132.85, 128.86, 128.56, 122.33, 81.79, 42.08, 35.04, 26.15, 24.58, 24.13. HRMS (ESI) Calcd for C 18 H 19 ClN2: [M+H] + = 299.1310. Found: 299.1318. Example 3: Preparation of Bis-Heterocyclic Compound 3c In a 10 mL reaction tube, γ, δ-unsaturated α-imidooxacid 1c (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 47.7 mg of the bicyclic heterocyclic compound 3c as a yellow solid with a yield of 81%.

[0043] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3c are: 1 H NMR (500 MHz, Chloroform- d ) δ 8.40 (d, J= 5.2 Hz, 2H), 7.35 –7.31 (m, 1H), 7.29 – 7.17 (m, 4H), 6.89 (dd, J = 7.7, 2.0 Hz, 1H), 4.30 (t, J = 7.7 Hz, 1H), 3.76 (s, 3H), 2.75 – 2.64 (m, 1H), 2.61 – 2.51 (m, 1H), 1.87 –1.75 (m, 1H), 1.40 (s, 3H), 1.39 – 1.34 (m, 1H), 1.32 (s, 3H). 13C{1H} NMR (126 MHz, Chloroform- d ) δ 172.87, 159.49, 156.58, 148.99,135.79, 129.29, 122.33, 120.21, 116.23, 112.45, 81.62, 55.22, 42.10, 35.19,26.21, 24.48, 23.97. HRMS (ESI) Calcd for C 19 H 22 N2O: [M+H] + = 295.1805. Found: 295.1816. Example 4: Preparation of Bis-Heterocyclic Compound 3d In a 10 mL reaction tube, γ, δ-unsaturated α-imidooxacid 1d (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the mixture was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 45.0 mg of the bicyclic heterocyclic compound 3d as a yellow solid in a 77% yield.

[0044] The above reaction equation is as follows: The relevant data of the biheterocyclic compound 3d are: 1 H NMR (600 MHz, Chloroform- d ) δ 8.49 (s, 2H), 7.42 (s, 2H), 7.34 (d, J = 4.7 Hz, 2H), 7.07 (s, 1H), 4.38 (t, J 1.40 (s, 3H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 173.57, 156.91, 149.00, 137.93,134.29, 132.19, 125.43, 122.48, 81.66, 42.28, 35.26, 26.46, 24.48, 23.96,21.23. HRMS (ESI) Calcd for C 20 H 24 N2: [M+H] + = 293.2012. Found: 293.2022. Example 5: Preparation of Bis-Heterocyclic Compound 3e In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1e (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 -(0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 45.0 mg of the bicyclic heterocyclic compound 3e as a yellow solid in a 74% yield.

[0045] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3e are: 1 H NMR (500 MHz, Chloroform- d ) δ 8.52 (d, J = 6.3 Hz, 2H), 7.77 –7.71 (m, 2H), 7.44 – 7.39 (m, 3H), 7.37 – 7.32 (m, 2H), 4.10 (dd, J = 8.6,6.9 Hz, 1H), 2.56 – 2.43 (m, 2H), 2.17 (dd, J = 12.9, 6.9 Hz, 1H), 2.12 –2.03 (m, 1H), 2.01 – 1.93 (m, 1H), 1.92 – 1.83 (m, 1H), 1.72 – 1.63 (m, 1H), 1.59 (dd, J = 12.9, 8.7 Hz, 1H), 1.48 (s, 3H), 1.42 (s, 3H). 13C{1H} NMR (126 MHz, Chloroform- d ) δ 178.16, 156.35, 149.21, 135.09,129.39, 128.29, 127.58, 122.47, 77.05, 54.83, 43.32, 41.24, 32.49, 31.34,26.31, 24.26, 16.05. HRMS (ESI) Calcd for C 21 H 24N2: [M+H] + = 305.2012. Found: 305.2022. Example 6: Preparation of Bis-Heterocyclic Compound 3f In a 10 mL reaction tube, γ, δ-unsaturated α-imidooxacid 1f (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 14.6 mg of the bicyclic heterocyclic compound 3f as a yellow liquid in a 32% yield.

[0046] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3f are: 1 H NMR (500 MHz, Chloroform- d ) δ 8.52 – 8.47 (m, 2H), 7.30 – 7.25 (m,2H), 4.16 (t, J = 7.5 Hz, 1H), 2.20 – 2.12 (m, 1H), 2.02 – 1.93 (m, 1H), 1.94– 1.85 (m, 1H), 1.77 – 1.68 (m, 1H), 1.41 (s, 3H), 1.35-1.29 (m, 4H), 0.89 –0.85 (m, 2H), 0.83 – 0.78 (m, 1H), 0.76 – 0.71 (m, 1H). 13C{1H} NMR (126 MHz, Chloroform- d) δ 179.87, 156.61, 149.09, 122.35,80.87, 41.81, 34.29, 26.19, 24.05, 23.82, 14.18, 7.09, 6.65. HRMS (ESI) Calcd for C 15 H 20 N2: [M+H] + = 229.1699. Found: 229.1694. Example 7: Preparation of Bis-Heterocyclic Compound 3g In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1 g (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1), yielding 29.7 mg of the biheterocyclic compound (3 g) as a white solid, with a yield of 56%.

[0047] The above reaction equation is as follows: The relevant data for 3g of the bicyclic heterocyclic compound are: 1 H NMR (600 MHz, Chloroform- d ) δ 8.60 (d, J = 4.5 Hz, 1H), 8.49 (d, J = 6.0 Hz, 2H), 8.14 (d, J = 7.9 Hz, 1H), 7.73 (td, J = 7.7, 1.6 Hz, 1H), 7.35(d, J= 6.1 Hz, 2H), 7.32 – 7.27 (m, 1H), 4.42 (t, J = 7.8 Hz, 1H), 2.94 –2.86 (m, 2H), 1.97 – 1.88 (m, 1H), 1.50-1.44 (m, 4H), 1.40 (s, 3H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 174.81, 157.12, 153.08, 149.03,148.81, 136.22, 124.64, 122.41, 121.92, 82.28, 42.10, 35.07, 26.02, 24.48,24.11. HRMS (ESI) Calcd for C 17 H 19 N3: [M+H] + = 266.1652. Found: 266.1662. Example 8: Preparation of Bis-Heterocyclic Compound 3h In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1h (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 34.5 mg of the bicyclic heterocyclic compound 3h as a white solid with a yield of 68%.

[0048] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3h are: 1H NMR (600 MHz, Chloroform- d ) δ 8.48 (d, J = 6.1 Hz, 2H), 7.52 (d, J = 1.4 Hz, 1H), 7.35 – 7.31 (m, 2H), 6.80 (d, J = 3.4 Hz, 1H), 6.46 (dd, J =3.4, 1.8 Hz, 1H), 4.39 (t, J = 7.6 Hz, 1H), 2.73 – 2.65 (m, 1H), 2.62 – 2.55(m, 1H), 1.88 – 1.81 (m, 1H), 1.49 (s, 3H), 1.45-1.38(m, 1H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 164.00, 156.80, 149.67, 148.87,144.75, 122.41, 113.55, 111.55, 81.98, 42.15, 35.05, 26.64, 24.17, 23.86. HRMS (ESI) Calcd for C 16 H 18 N2O: [M+H] + = 255.1492. Found: 255.1500. Example 9: Preparation of Bis-Heterocyclic Compound 3i In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1i (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2a (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 -(0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 38.9 mg of the bicyclic heterocyclic compound 3i as a white solid in a 72% yield.

[0049] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3i are: 1 H NMR (600 MHz, Chloroform- d ) δ 8.47 (d, J = 5.6 Hz, 2H), 7.40 (d, J = 5.0 Hz, 1H), 7.30 (d, J = 6.2 Hz, 2H), 7.23 (d, J = 3.5 Hz, 1H), 7.04 (dd, J = 5.0, 3.7 Hz, 1H), 4.35 (t, J = 7.5 Hz, 1H), 2.79 – 2.70 (m, 1H), 2.62 –2.55 (m, 1H), 1.94 – 1.86 (m, 1H), 1.49-1.43 (m, 4H), 1.40 (s, 3H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 167.61, 156.33, 149.01, 139.36,129.10, 129.04, 127.41, 122.46, 81.58, 42.22, 35.72, 26.36, 24.80, 24.16. HRMS (ESI) Calcd for C 16 H 18 N2S: [M+H] + = 271.1263. Found: 271.1275. Example 10: Preparation of Bis-Heterocyclic Compound 3j In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1a (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2b (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 47.1 mg of the bicyclic heterocyclic compound 3j as a yellow liquid with an 80% yield.

[0050] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3j are: 1 H NMR (500 MHz, Chloroform- d ) δ 8.04 (d, J = 5.5 Hz, 1H), 7.83 (dd, J = 7.7, 1.8 Hz, 2H), 7.45 – 7.36 (m, 3H), 6.95 (dd, J = 5.6, 1.5 Hz, 1H), 6.77 (s, 1H), 4.38 (t, J = 7.7 Hz, 1H), 3.91 (s, 3H), 2.86 – 2.77 (m, 1H), 2.77 – 2.68 (m, 1H), 1.94 – 1.85 (m, 1H), 1.52-1.44 (m, 4H), 1.34 (s, 3H). 13C{1H} NMR (126 MHz, Chloroform- d) δ 172.95, 164.40, 159.73, 146.04,134.60, 130.34, 128.35, 127.62, 115.98, 109.00, 81.71, 53.27, 42.22, 35.21,26.74, 24.56, 23.43. HRMS (ESI) Calcd for C 19 H 22 N2O: [M+H] + = 295.1805. Found: 295.1814. Example 11: Preparation of Bis-Heterocyclic Compound 3k In a 10 mL reaction tube, γ, δ-unsaturated α-imidooxacid 1a (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2c (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 28.8 mg of the bicyclic heterocyclic compound 3k as a yellow solid in a 51% yield.

[0051] The above reaction equation is as follows: The relevant data of the bicyclic compound 3k are: 1 H NMR (500 MHz, Chloroform- d ) δ 8.37 (d, J = 4.2 Hz, 1H), 8.36 –8.31 (m, 1H), 7.85 – 7.80 (m, 2H), 7.45 – 7.37 (m, 3H), 7.28 (dd, J = 7.1,5.2 Hz, 1H), 4.81 (t, J= 7.8 Hz, 1H), 2.93 – 2.86 (m, 2H), 1.97 – 1.87 (m,1H), 1.60-1.52(m, 4H), 1.31 (s, 3H). 13C{1H} NMR (126 MHz, Chloroform- d ) δ 172.93, 158.83 (d, J = 256.3 Hz),145.55 (d, J = 4.7 Hz), 143.52 (d, J = 9.1 Hz), 138.89 (d, J = 28.6 Hz),134.38, 130.50, 128.39, 127.72, 123.16 (d, J = 3.1 Hz), 78.85 (d, J = 4.2Hz), 42.18 (d, J = 4.0 Hz), 35.42, 24.91 (d, J = 3.5 Hz), 24.51, 22.12 (d, J = 2.6 Hz). 19 F NMR (565 MHz, Chloroform- d ) δ -123.54. HRMS (ESI) Calcd for C 18 H 19 FN2: [M+H] + = 283.1605. Found: 283.1615. Example 12: Preparation of Bis-Heterocyclic Compound 31 In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1a (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2d (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 -(0.0025 mmol, 2.5 mol%), resulting in a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After completion of the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 47.3 mg of the bicyclic heterocyclic compound 31 as a yellow liquid, with a yield of 81%.

[0052] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3l are: 1 H NMR (600 MHz, Chloroform- d ) δ 7.81 (dd, J = 7.9, 1.7 Hz, 2H), 7.44– 7.36 (m, 3H), 6.98 (s, 2H), 4.38 (t, J = 7.6 Hz, 1H), 2.83 – 2.74 (m, 1H), 2.67 – 2.59 (m, 1H), 2.47 (s, 6H), 1.92 – 1.84 (m, 1H), 1.49-1.43 (m, 4H), 1.35 (s, 3H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 172.96, 157.07, 156.91, 134.59,130.31, 128.33, 127.55, 118.90, 81.79, 42.04, 35.17, 26.64, 24.55, 24.52,23.72. HRMS (ESI) Calcd for C 20 H 24 N2: [M+H] + = 293.2012. Found: 293.2022. Example 13: Preparation of Bis-Heterocyclic Compound 3m In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1a (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2e (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 54.7 mg of the bicyclic heterocyclic compound 3m as a yellow liquid with a yield of 67%.

[0053] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3m are: 1 H NMR (500 MHz, Chloroform- d ) δ 8.61 (d, J = 5.2 Hz, 1H), 7.96 (d, J = 8.1 Hz, 2H), 7.92 – 7.86 (m, 2H), 7.80 (s, 1H), 7.63 (d, J = 8.2 Hz, 2H),7.50 – 7.39 (m, 3H), 7.34 (dd, J = 5.3, 1.8 Hz, 1H), 4.45 (t, J = 7.6 Hz,1H), 2.88 – 2.77 (m, 1H), 2.69 – 2.59 (m, 1H), 2.07 – 1.96 (m, 1H), 1.57-1.48(m, 7H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 173.29, 157.26, 155.49, 149.46,143.26, 134.37, 130.64, 130.47 (q,J = 32.3 Hz), 128.49, 127.72, 127.32,125.51 (q, J = 3.7 Hz), 124.24 (q, J = 272.1 Hz), 121.73, 120.27, 81.94, 42.50, 35.14, 26.08, 25.32, 24.73. 19 F NMR (565 MHz, Chloroform- d ) δ -62.5. HRMS (ESI) Calcd for C 25 H 23 F3N2: [M+H] + = 409.1886 . Found: 409.1898. Example 14: Preparation of Bis-Heterocyclic Compound 3n In a 10 mL reaction tube, γ, δ-unsaturated α-iminooxacid 1a (0.2 mmol, 1.0 equiv.), N-aminopyridinium salt 2f (0.3 mmol, 1.5 equiv.), sodium acetate (0.4 mmol, 2.0 equiv.) and Mes-Acr were added. + ClO4 - (0.0025 mmol, 2.5 mol%) to obtain a mixed solution. Under nitrogen protection, 2 mL of acetonitrile was added to the resulting mixture, followed by magnetic stirring at room temperature for 16 h under blue light irradiation (wavelength 456 nm, light intensity 40 W). After the reaction, the resulting reaction solution was extracted with ethyl acetate (3.125 mL). The combined organic phases were washed with saturated brine (1.130 mL), dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. The concentrated crude product was separated and purified by silica gel column chromatography (eluent, dichloromethane:methanol = 60:1) to obtain 48.9 mg of the biheterocyclic compound 3n as a yellow liquid in a 65% yield.

[0054] The above reaction equation is as follows: The relevant data of the bicyclic heterocyclic compound 3n are: 1 H NMR (600 MHz, Chloroform- d ) δ 8.58 (d, J = 5.3 Hz, 1H), 7.87 (td,J = 8.8, 6.8 Hz, 1H), 7.85 – 7.81 (m, 2H), 7.77 (s, 1H), 7.45 – 7.36 (m, 3H),7.31 (dd, J = 5.3, 1.8 Hz, 1H), 6.95 (td, J = 8.4, 2.5 Hz, 1H), 6.92 – 6.85(m, 1H), 4.43 (t, J = 7.7 Hz, 1H), 2.86 – 2.77 (m, 1H), 2.75 – 2.66 (m, 1H),2.01 – 1.92 (m, 1H), 1.56-1.49 (m, 4H), 1.46 (s, 3H). 13C{1H} NMR (151 MHz, Chloroform- d ) δ 173.15, 162.95 (dd, J = 250.5, 12.0Hz), 160.38 (dd, J = 252.2, 12.0 Hz), 157.05, 152.21 (d, J = 2.7 Hz), 149.16,134.40, 132.15 (dd, J = 9.6, 4.5 Hz), 130.41, 128.32, 127.63, 124.27 (dd, J =11.8, 3.8 Hz), 122.99 (d, J = 8.5 Hz), 121.25, 111.65 (dd, J = 21.0, 3.6 Hz),104.24 (dd, J = 26.9, 25.4 Hz), 81.74, 42.30, 35.15, 26.26, 24.58, 24.22. 19 F NMR (565 MHz, Chloroform- d ) δ -109.73 (d, J = 8.5 Hz), -112.78 (d, J = 8.4 Hz). HRMS (ESI) Calcd for C24 H 22 F2N2: [M+H] + = 377.1824. Found: 377.1836. Experiment 1: Cytotoxicity assay of different compounds In order to explore the anti-tumor effects of compounds 3a-3n obtained in Examples 1-14 of the present invention, the following tests were performed using human colon cancer cells HCT-116 as experimental subjects: Cell culture: HCT-116 (human colon cancer cells, purchased from the Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) cells were cultured at 37°C in a 5% CO2 atmosphere in high-glucose Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Catalog No. C11995500BT) supplemented with 10% fetal bovine serum (FBS) (Vazyme, Catalog No. F101-01) and 1% penicillin-streptomycin solution (Biosharp, Catalog No. BL505A). Cells were passaged when they reached 70-80% confluency, with experimental passages limited to five.

[0055] Cell treatment: HCT-116 cells in the logarithmic growth phase were selected, the cell suspension was counted using a cell counter, and the concentration was adjusted to 5×10 4 Cells were seeded in a 96-well plate (3000 cells / well) at a volume of 100 μL per well and incubated overnight to ensure adequate cell attachment. The culture medium was then replaced with fresh medium containing various final concentrations of the compound (0, 0.015, 0.046, 0.14, 0.41, 1.23, 3.7, 11.11, 33.33, and 100 mM / μM) (the anti-tumor drug cisplatin was also used as a positive control) and cultured for an additional 72 hours. Subsequently, 20 μL of CCK-8 solution (Beyotime, Shanghai Beyotime Biotechnology Co., Ltd.) was added to each well and incubated at 37°C for 2 hours. After incubation, the optical density (OD) of each well was measured at 450 nm using a microplate reader (Tecan Infinite M1000 Pro).

[0056] The relative cell viability of each sample was normalized using a DMSO control. IC50 is used to measure the level of inhibition or activity of a compound or drug on a biological system. The present invention calculates the IC50 value of a compound on tumor cells by cell viability.

[0057] Cell viability (%) = OD450 of sample-treated group / average OD450 of blank control group × 100%.

[0058] The half inhibitory concentration IC of each compound on HCT-116 cells 50 The statistical results are shown in Table 1. As shown in Table 1, the compounds obtained in Examples 1-14 have good biological activity and can effectively inhibit the vitality of colon cancer cells.

[0059] Table 1 IC values of each compound against HCT 116 50 The above description is merely a preferred embodiment of the present invention and does not constitute any form of limitation to the present invention. Although the present invention has been disclosed as a preferred embodiment as above, it is not intended to limit the present invention. Any technician familiar with this profession can make slight changes or modifications to equivalent embodiments of the methods and technical contents disclosed above without departing from the scope of the technical solution of the present invention. However, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A compound represented by formula III or a pharmaceutically acceptable salt thereof: in, R 1 Refers to any of alkyl and aryl groups; R 2 Refers to either fluorine or hydrogen; R 3 、R 4 Each refers to any of aryl, methoxy, methyl, and hydrogen.

2. The method for preparing the compound of formula III or a pharmaceutically acceptable salt thereof according to claim 1, wherein: The method comprises the following steps: S1: γ,δ-unsaturated α-imidooxo acid, N-aminopyridinium salt, sodium acetate and Mes-Acr + ClO4 - mixing to obtain a mixed solution; S2: Under an inert atmosphere, adding an organic solvent to the obtained mixed solution and stirring the mixture under blue light irradiation to obtain a reaction solution; S3: The obtained reaction solution is concentrated and purified to obtain the compound represented by formula III.

3. The preparation method according to claim 2, characterized in that In step S2, the inert atmosphere is selected from any one of nitrogen and argon.

4. The preparation method according to claim 2, characterized in that In step S2, the organic solvent is selected from any one of acetonitrile and 1,2-dichloroethane.

5. The preparation method according to claim 2, characterized in that In step S2, the blue light irradiation conditions are: wavelength 424-525nm, light intensity 10-40W.

6. The preparation method according to claim 2, characterized in that In step S2, the stirring reaction conditions are: room temperature, time 8-24h.

7. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (1) The compound of formula III according to claim 1 or a pharmaceutically acceptable salt thereof; and (2) Pharmaceutically acceptable carriers and / or excipients.

8. Use of the compound of formula III or a pharmaceutically acceptable salt thereof according to claim 1 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for treating and / or preventing diseases associated with abnormal cell activity.

9. The use according to claim 8, characterized in that The disease associated with abnormal cell activity is selected from tumors.

10. The use according to claim 8, characterized in that The tumor is colorectal cancer.

Images