Photosensitizer based on quinoline-malononitrile mother nucleus as well as preparation method and application of photosensitizer

Abstract

The invention discloses a quinoline-malononitrile mother nucleus-based photosensitizer as well as a preparation method and application thereof. The structure of the photosensitizer is characterized in that a quinoline nitrile fragment is taken as an electron acceptor, triphenylamine and substituted triphenylamine fragments are taken as electron donors, and a carbon-carbon double bond or thiophene heterocyclic ring is taken as a pi bridge to connect the electron acceptor and the electron donor. Meanwhile, the invention further discloses a preparation method of the photosensitizer and potential biological application of killing tumor cells through the photodynamic effect.

Description

Technical Field

[0001] The present invention belongs to the field of tumor photodynamic therapy, and particularly relates to a photosensitizer based on a quinoline-malononitrile nucleus, a preparation method thereof, and an application thereof. Background Art

[0002] Photodynamic Therapy (PDT) is a new treatment method that irradiates tumor tissues with an excitation light source of a specific wavelength, activates photosensitizers (PS) selectively aggregated in the tumor tissues, and triggers a photochemical reaction to generate cytotoxic ROS, such as peroxides (O2 2- ), superoxides (O2 - ), hydroxyl radicals (·OH), and singlet oxygen ( 1 O2), etc., causing apoptosis or necrosis of tumor cells, thereby playing a role in tumor treatment.

[0003] Compared with traditional cancer treatment methods such as surgery, chemotherapy, and radiotherapy, PDT has the advantages of non-invasiveness, high selectivity, and low side effects. At present, PDT has been widely used in the clinical treatment of superficial bladder cancer, Barrett esophagus cancer, and skin cancer. At the same time, PDT can also be used as an adjuvant treatment method to enhance the treatment effect in combination with traditional cancer treatment methods. In addition, new application scenarios of PDT are constantly being developed. For example, in the fields of antibacterial and antiviral, PDT therapy is also constantly evolving.

[0004] The efficacy of PDT is affected by three factors: (1) photosensitizer; (2) tissue oxygen content; (3) excitation light source. Among them, the wavelength of the excitation light source is generally determined by the photophysical properties of the photosensitizer, and the tissue oxygen content is related to the tumor microenvironment. The core of photodynamic therapy is the photosensitizer. Developing photosensitizers with new structural types or different action mechanisms to solve the deficiencies of existing photosensitizers is the focus of research in this field. Aggregation-induced emission type photosensitizers can still emit strong fluorescence and efficiently generate reactive oxygen species under light excitation in the molecular aggregation state, solving the problem of aggregation-induced quenching effect of traditional photosensitizers, and are easy to realize imaging-guided photodynamic therapy, and have attracted much attention in recent years. Summary of the Invention

[0005] One of the purposes of the present invention is to provide a photosensitizer based on a quinoline-malononitrile nucleus, a preparation method thereof, and an application thereof. The photosensitizer is a D (Donor, D)-π (π-bridge)-A (Accepter, A) type photosensitizer molecule. This type of molecule uses a quinoline nitrile fragment as the photosensitizer electron acceptor, a triphenylamine and substituted triphenylamine fragment as the photosensitizer electron donor, and a carbon-carbon double bond or a thiophene heterocycle as the π-bridge to connect the electron acceptor and the donor to construct the photosensitizer molecule.

[0006] The technical solution is as follows:

[0007] A photosensitizer based on a quinoline-malononitrile core, which is a compound represented by the general formula (I):

[0008]

[0009] Wherein R1 is H, OH, OTs, SCH3, I, Br, N3, NH2,

[0010] R2 is H, CH3, OCH3 or F;

[0011] Linker is

[0012] Furthermore:

[0013] R1 is I, Br, NH2, OH,

[0014] R2 is H, CH3 or OCH3.

[0015] Furthermore, the compound represented by the general formula (I) or its pharmaceutically acceptable salt or its stereoisomer or its prodrug is selected from the following compounds:

[0016] (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-ethylquinolin-4(1H)-ylidene)malononitrile, namely QM01;

[0017] (E)-2-(2-(4-(Diphenylamino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM02;

[0018] (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM03;

[0019] (E)-2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM04;

[0020] (E)-2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM05;

[0021] (E)-2-(2-(2-(5-(4-(Diphenylamino)phenyl)thiophen-2-yl)vinyl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM06;

[0022] (E)-2-(2-(2-(5-(4-(Di(p-tolyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM07;

[0023] (E)-2-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM08;

[0024] (E)-2-(2-(4-(Diphenylamino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM09;

[0025] (E)-2-(2-(4-(Di(p-tolyl)amino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM10;

[0026] (E)-2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM11;

[0027] (E)-2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM12; (E)-2-(2-(2-(5-(4-(Diphenylamino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM13;

[0028] (E)-2-(2-(2-(5-(4-(Di(p-tolyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM14;

[0029] (E)-2-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM15;

[0030] (E)-2-(2-(2-(5-(4-(Bis(4-fluorophenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM16;

[0031] (E)-(2-(4-(Diphenylamino)-2-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate, namely QM17;

[0032] (E)-(2-(2-(4-(Di-p-tolylamino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfate, namely QM18;

[0033] (E)-(2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate, namely QM19;

[0034] (E)-(2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate, namely QM20;

[0035] (E)-(2-(4-(Dicyanovinyl)-2-(2-(5-(4-(diphenylamino)phenyl)thiophen-2-yl)vinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate, namely QM21;

[0036] (E)-(2-(2-(5-(4-(Di-p-tolylamino)phenyl)thiophen-2-yl)vinyl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate, namely QM22;

[0037] (E)-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate, namely QM23;

[0038] (E)-2-(2-(4-(Di-p-tolylamino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl 4-methylbenzenesulfonate, namely QM24;

[0039] (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-(2-iodoethyl)quinolin-4(1H)-ylidene)propanedinitrile, namely QM25;

[0040] (E)-2-(1-(2-Bromoethyl)-2-(4-(di-p-toluidino)styryl)quinolin-4(1H)-ylidene)propanedinitrile, namely QM26;

[0041] (E)-2-(1-(2-Azidoethyl)-2-(4-(di-p-toluidino)styryl)quinolin-4(1H)-ylidene)propanedinitrile, namely QM27;

[0042] (E)-2-(1-(2-Aminoethyl)-2-(4-(di-p-tolylamino)styryl)quinolin-4(1H)-ylidene)malononitrile, namely QM28;

[0043] (E)-(4-((2-(4-(Di-p-tolylamino)styryl)-4-(dicyanomethyl)quinolin-1(4H)-yl)ethyl)amino)-4-oxobutyl)triphenylphosphonium, namely QM29;

[0044] (E)-1-(4-((2-(4-(Di-p-tolylamino)styryl)-4-(dicyanomethyl)quinolin-1(4H)-yl)ethyl)amino)-4-oxobutyl)pyridin-1-ium, namely QM30.

[0045] Another object of the present invention is to provide a preparation method of the photosensitizer based on the quinoline-malononitrile nucleus, which is achieved through the following steps:

[0046] i) Using 2-methylquinoline and bromide as starting materials, using toluene as a solvent, reacting at 110 °C to obtain the corresponding quinolinium salt;

[0047] ii) Under the condition that the quinolinium salt uses sodium ethoxide as a base and ethanol as a solvent, reacting with malononitrile to obtain the corresponding quinoline-malononitrile nuclei Q1 and Q2; Q2 is obtained by OTs substitution and reaction with sodium methanethiolate to obtain Q3;

[0048] iii) Q1, Q2, and Q3 are subjected to Knoevenagel condensation reaction with triphenylamine containing a mono-substituted aldehyde group to obtain the photosensitizer based on the quinoline-malononitrile nucleus.

[0049] Furthermore:

[0050] The photosensitizers QM02-QM08 containing a methylthio substitution react with methyl iodide and silver tetrafluoroborate to obtain the corresponding sulfonium-substituted photosensitizers QM17-QM23;

[0051] The hydroxyl group of the photosensitizer QM10 containing a hydroxyl substitution undergoes p-toluenesulfonylation to obtain QM24, and subsequently, through further substitution reactions, bromine, iodine, and azide-substituted photosensitizer molecules QM25, QM26, and QM27 are obtained;

[0052] The azide-substituted photosensitizer molecule QM27 is reduced to obtain an amino-substituted photosensitizer molecule QM28, and further through a condensation reaction, a pyridinium salt and triphenylphosphonium are introduced to obtain the corresponding QM29 and QM30.

[0053] Another object of the present invention is to provide the application of the photosensitizer based on the quinoline-malononitrile nucleus in the preparation of drugs for treating or preventing tumors. The compound, or its pharmaceutically acceptable salt, or its stereoisomer, or its prodrug kills tumor cells through photodynamic therapy and exerts an anti-tumor effect. Preferably, the tumor cells are murine triple-negative breast cancer cells and human cervical cancer cells Hela.

[0054] The photosensitizer based on the quinoline-malononitrile nucleus described in the present invention has the ability to generate type I reactive oxygen species and is a class of oxygen-independent photosensitizers.

[0055] The photosensitizer based on the quinoline-malononitrile nucleus described in the present invention has an endoplasmic reticulum targeting function.

[0056] The photosensitizer based on the quinoline-malononitrile nucleus described in the present invention has the property of aggregation-induced fluorescence enhancement and can achieve autofluorescence imaging in the near-infrared region.

[0057] The technical solutions provided by the embodiments of the present application may include the following beneficial effects:

[0058] 1. Compared with the first-generation to third-generation photosensitizers, the photosensitizer based on the quinoline-malononitrile nucleus provided by the present invention is simple in synthesis and low in cost.

[0059] 2. The photosensitizer based on the quinoline-malononitrile nucleus provided by the present invention has a large molar extinction coefficient, a large Stokes shift, and the property of aggregation-induced fluorescence enhancement. Compared with traditional photosensitizers, it has a higher reactive oxygen species generation efficiency and can achieve autofluorescence imaging in the near-infrared region at the same time.

[0060] 3. The photosensitizer based on the quinoline-malononitrile nucleus provided by the present invention has a wide therapeutic window, that is, this type of photosensitizer shows high photodynamic activity and low dark cytotoxicity to cells; it has good photodynamic therapy effects on murine triple-negative breast cancer cells 4T1 and human cervical cancer cells Hela.

[0061] 4. The photosensitizer based on the quinoline-malononitrile nucleus provided by the present invention can effectively inhibit the tumor growth of breast cancer-bearing mice and has a small impact on the weight loss of mice.

[0062] 5. The photosensitizer based on the quinoline-malononitrile nucleus provided by the present invention has an endoplasmic reticulum targeting function.

[0063] In summary, the photosensitizers provided by the present invention have a large Stokes shift, a high in vitro and in vivo reactive oxygen species generation efficiency, and aggregation-induced fluorescence enhancement properties; cell results show that such photosensitizers have high photodynamic activity (0.6 - 5 μM) and low dark toxicity (>100 μM) against different types of cancer cells, with a good therapeutic window; such photosensitizers also have an endoplasmic reticulum localization function, and it has also been confirmed in a mouse breast cancer model that such photosensitizers can effectively inhibit the tumor growth of tumor-bearing mice with breast cancer, and have the potential to be further developed into new anti-tumor photosensitizers.

[0064] It should be understood that the above general description and the following detailed description are only exemplary and explanatory, and cannot limit this application. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] The drawings herein are incorporated into the specification and constitute a part of this specification, showing embodiments consistent with this application, and are used together with the specification to explain the principles of this application.

[0066] Figure 1 Schematic diagram of the ultraviolet-visible light absorption spectrum and fluorescence emission spectrum of some photosensitizer molecules (concentration 20 μM).

[0067] Figure 2 Schematic diagram of the fluorescence emission spectrum of some photosensitizer molecules in water / dimethyl sulfoxide solutions with different ratios (concentration 20 μM).

[0068] Figure 3 Images of photosensitizers under laser irradiation (taking QM10 and QM18 as examples), where (a) is the fluorescence intensity comparison diagram of QM18 in dimethyl sulfoxide solutions with different water contents, (b) is the fluorescence intensity comparison diagram of QM10 in dimethyl sulfoxide solutions with different water contents, (c) is the morphological diagram of QM18 under 365 nm ultraviolet lamp irradiation, and (d) is the morphological diagram of QM10 under 365 nm ultraviolet lamp irradiation.

[0069] Figure 4 Schematic diagram for evaluating the reactive oxygen species generation ability of some photosensitizer molecules in an aqueous solution containing a reactive oxygen indicator (DCFH) after continuous light irradiation, where (a) is the schematic diagram of the irradiation wavelength and relative fluorescence intensity change of QM01, QM03, QM09 - QM14, (b) is the schematic diagram of the irradiation wavelength and relative fluorescence intensity change of QM15 - QM22, and (c) is the schematic diagram of the irradiation wavelength and relative fluorescence intensity change of QM23 - QM30.

[0070] Figure 5Schematic diagram for evaluating the ability of some photosensitizer molecules to generate reactive oxygen species under continuous light irradiation in an aqueous solution containing a singlet oxygen indicator (ABDA), where (a) is a schematic diagram of the light irradiation wavelength and absorbance change of QM09 - QM16, and (b) is a schematic diagram of the light irradiation wavelength and absorbance change of QM17 - QM23 and ABDA.

[0071] Figure 6 Evaluation of the ability of some photosensitizer molecules to generate reactive oxygen species under continuous light irradiation in an aqueous solution containing a hydroxyl radical indicator (HPF), where (a) is a schematic diagram of the light irradiation wavelength and absorbance change of QM09 - QM15 and HPF, and (b) is a schematic diagram of the light irradiation wavelength and absorbance change of QM17 - QM23.

[0072] Figure 7 Co - localization map of the endoplasmic reticulum of 4T1 cells by photosensitizer QM10, where (a) is the laser confocal microscopy image of QM10 in Hela cells, (b) is the laser confocal microscopy image of the endoplasmic reticulum probe in Hela cells, (c) is the superimposed image of (a) and (b), (d) is the bright - field image of the laser confocal microscopy of Hela cells, and (e) is the correlation distribution map of the superimposed image of (a) and (b).

[0073] Figure 8 Bar chart of the phototoxicity and dark toxicity of photosensitizers to 4T1 cells.

[0074] Figure 9 In - vivo photodynamic effect diagram of photosensitizer QM10, where A is the small - animal in - vivo imaging diagram of mice after intratumoral injection of QM10 over time, B is the body weight curve diagram of mice over time after different treatment regimens, C is the tumor volume diagram over time after different treatment regimens, D is the mouse tumor diagram separated by dissection after different treatment regimens, E is the mouse tumor mass diagram separated by dissection after different treatment regimens, F is the appearance of mice after being treated with QM10 and Ce6 and sacrificed, and G is the H&E staining result diagram of the main organs of mice after different treatment regimens. Detailed implementation mode

[0075] Here, the exemplary embodiments will be described in detail, and the examples are shown in the accompanying drawings. When the following description refers to the drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The implementation manners described in the following exemplary embodiments do not represent all the implementation manners consistent with the present application.

[0076] The terms used in this application are for the purpose of describing specific embodiments only and are not intended to limit this application. The singular forms "a", "the", and "said" used in this application and the appended claims are also intended to include the plural forms unless the context clearly dictates otherwise. It should also be understood that the term "and / or" as used herein refers to and encompasses any and all possible combinations of one or more of the associated listed items.

[0077] It should be understood that although the terms first, second, third, etc. may be used in this application to describe various information, such information should not be limited to these terms. These terms are only used to distinguish information of the same type from each other. For example, without departing from the scope of this application, the first information may also be referred to as the second information, and similarly, the second information may also be referred to as the first information. Depending on the context, the word "if" as used herein may be interpreted as "when" or "while" or "in response to a determination".

[0078] Example 1:

[0079] (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-ethylquinolin-4(1H)-ylidene)malononitrile

[0080] QM01

[0081]

[0082] 2-(1-Ethyl-2-methylquinolin-4(1H)-ylidene)malononitrile (5 mmol, 1.17 g), 4-(di-p-toluidino)benzaldehyde (7.5 mmol, 2.26 g), 1.5 mL of piperidine and 50 mL of acetonitrile were added to a 100 mL pear-shaped flask, and the mixture was stirred and reacted at 75 °C in an oil bath for 12 h, monitored by TLC; after the reaction was completed, the solvent was removed by distillation under reduced pressure, and the residue was slurried with methanol and filtered to obtain 4.12 g of the target compound. A red solid with a yield of 82%. 1 H NMR (400 MHz, DMSO-d6) δ 8.89 (dd, J = 8.6, 0.8 Hz, 1H), 8.06 (d, J = 9.0 Hz, 1H), 7.91 (t, J = 7.6 Hz, 1H), 7.64 (d, J = 8.8 Hz, 2H), 7.61 - 7.54 (m, 1H), 7.32 (d, J = 2.8 Hz, 2H), 7.16 (d, J = 8.2 Hz, 4H), 6.98 (d, J = 8.0 Hz, 5H), 6.86 (d, J = 8.7 Hz, 2H), 4.55 (dd, J = 13.6, 6.6 Hz, 2H), 2.29 (s, 6H), 1.39 (t, J = 7.0 Hz, 3H). 1313C NMR (100 MHz, DMSO-d6) δ 152.57, 150.05, 149.75, 144.41, 142.71, 140.04, 138.38, 134.16, 133.92, 130.74, 129.96, 128.08, 125.70, 125.63, 121.14, 120.58, 118.61, 117.97, 107.05, 46.88, 20.95, 14.16. HRMS (ESI) m / z: [M+H] + calcd for C 36 H 31 N4 + 519.2543, found: 519.2540.

[0083] Example 2:

[0084] (E)-2-(2-(4-(Di-p-tolylamino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile

[0085] QM03

[0086]

[0087] The synthesis method was the same as that of Example 1, except that 2-(1-ethyl-2-methylquinolin-4(1H)-ylidene)malononitrile was replaced by 2-(2-methyl-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile. Red solid, yield 61%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.91 (dd, J = 8.4, 1.2 Hz, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.94 - 7.89 (m, 1H), 7.67 - 7.57 (m, 3H), 7.42 (d, J = 15.6 Hz, 1H), 7.34 (d, J = 15.6 Hz, 1H), 7.17 (d, J = 8.0 Hz, 4H), 7.00 (d, J = 8.0 Hz, 5H), 6.87 (d, J = 8.8 Hz, 2H), 4.72 (t, J = 7.4 Hz, 2H), 3.01 - 2.91 (m, 2H), 2.29 (s, 6H), 2.12 (s, 3H). 1313C NMR(100MHz,DMSO-d6)δ152.24,150.10,149.27,143.88,139.53,138.00,133.72,133.50,130.26,129.92,129.33,125.30,123.77,123.69,120.55,119.94,118.21,117.86,106.58,47.68,47.25,31.21,20.47,15.01.HRMS(ESI)m / z:[M+H] + calcd for C 37 H 33 N4S + 565.2420,found:565.2421.

[0088] Example 3:

[0089] (E)-(2-(2-(4-(Di-p-tolylamino)styryl)-4-(dicyanomethyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate

[0090] QM18

[0091]

[0092] QM03(1mmol,0.56g),methyl iodide(1.2mmol,0.17g),silver tetrafluoroborate(1.2mmol,0.23g) and 20 mL of dichloromethane were added to a 50 mL pear-shaped flask and stirred at room temperature for 48 h,monitored by TLC;after the reaction was completed,the solvent was removed by distillation under reduced pressure and purified by column chromatography to obtain 0.21 g of the target compound.Dark red solid,yield 31%. 1 1H NMR(400MHz,DMSO-d6)δ8.94(d,J = 7.6Hz,1H),8.13(d,J = 8.8Hz,1H),7.97(t,J = 8.0Hz,1H),7.72(d,J = 8.8Hz,2H),7.68 - 7.60(m,1H),7.42 - 7.40(m,1H),7.18(d,J = 8.2Hz,4H),7.04(s,1H),7.00(d,J = 8.2Hz,4H),6.87(d,J = 8.8Hz,2H),5.07(t,J = 7.0Hz,2H),3.81(t,J = 7.0Hz,2H),2.98(s,6H),2.30(s,6H). 1313C NMR(100MHz,DMSO-d6)δ149.86,149.46,143.82,140.51,137.98,133.93,133.66,130.31,129.73,129.34,127.35,125.38,125.31,121.53,120.60,120.17,119.95,119.74,118.02,117.13,106.71,48.34,25.15,20.48,15.03.HRMS(ESI)m / z:[M] + calcd for C 38 H 35 N4S + 579.2577,found:579.2574.

[0093] Example 4:

[0094] (E)-2-(2-(4-(Di-p-tolylamino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile

[0095] QM10

[0096]

[0097] The synthesis method was the same as that of Example 1, except that 2-(1-ethyl-2-methylquinolin-4(1H)-ylidene)malononitrile was replaced by 2-(1-(2-hydroxyethyl)-2-methylquinolin-4(1H)-ylidene)malononitrile. Red solid, with a yield of 79%. 1 1H NMR(400MHz,DMSO-d6)δ8.89(d,J = 8.4Hz,1H),8.07(d,J = 9.0Hz,1H),7.88(t,J = 7.8Hz,1H),7.61 - 7.55(m,3H),7.49(d,J = 15.6Hz,1H),7.28(d,J = 15.6Hz,1H),7.16(d,J = 8.2Hz,4H),7.05 - 6.95(m,5H),6.88(d,J = 8.6Hz,2H),5.19(t,J = 5.6Hz,1H),4.61(t,J = 4.6Hz,2H),3.85(q,J = 5.4Hz,2H),2.29(s,6H). 1313C NMR(100MHz, DMSO-d6) δ 152.19, 150.79, 149.18, 143.96, 139.03, 138.51, 133.44, 133.39, 130.25, 129.27, 127.78, 125.17, 125.03, 120.69, 120.24, 118.89, 118.58, 106.57, 58.90, 50.48, 46.47, 20.47. HRMS(ESI) m / z: [M+H] + calcd for C 36 H 31 N4O + 535.2492, found: 535.2499.

[0098] Example 5:

[0099] (E)-2-(2-(4-(Di-p-tolylamino)styryl)-4-(dicyanomethylene)quinolin-1(4H)-yl)ethyl 4-methylbenzenesulfonate

[0100] QM24

[0101]

[0102] QM10 (5 mmol, 2.67 g), p-toluenesulfonyl chloride (7.5 mmol, 1.42 g), 4-dimethylaminopyridine (0.5 mmol, 0.061 g) and 20 mL of dichloromethane were added to a 50 mL pear-shaped flask; after adding triethylamine (10 mmol) to the pear-shaped flask, it was stirred at room temperature for 4 h and monitored by TLC; after the reaction was completed, the solvent was removed by distillation under reduced pressure and purified by column chromatography to obtain 1.79 g of the target compound. Red solid, yield 52%. 1 1H NMR(400MHz, DMSO-d6) δ 8.89(d, J = 8.4Hz, 1H), 8.07(d, J = 9.0Hz, 1H), 7.88(t, J = 7.8Hz, 1H), 7.61 - 7.55(m, 3H), 7.49(d, J = 15.6Hz, 1H), 7.28(d, J = 15.6Hz, 1H), 7.16(d, J = 8.2Hz, 4H), 7.05 - 6.95(m, 5H), 6.88(d, J = 8.6Hz, 2H), 5.19(t, J = 5.6Hz, 1H), 4.61(t, J = 4.6Hz, 2H), 3.85(q, J = 5.4Hz, 2H), 2.29(s, 6H). HRMS(ESI) m / z: [M+H] + calcd for C 43 H 37 N4O3S+ 689.2581, found: 689.2588.

[0103] Example 6:

[0104] (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-(2-iodoethyl)quinolin-4(1H)-ylidene)malononitrile

[0105] QM25

[0106]

[0107] Add QM24 (1 mmol, 0.69 g), sodium iodide (1.5 mmol, 0.23 g) and 10 mL of acetone into a 50 mL pear-shaped flask; stir at room temperature for 12 h and monitor by TLC; after the reaction is completed, remove the solvent by distillation under reduced pressure, filter by suction and wash the solid with water to obtain 0.457 g of the target compound. Dark red solid, yield 71%. 1 H NMR (400 MHz, DMSO) δ 8.92 (d, J = 8.4 Hz, 1H), 8.07 - 7.86 (m, 2H), 7.66 - 7.62 (m, 3H), 7.38 - 7.35 (m, 2H), 7.18 (d, J = 7.5 Hz, 4H), 7.04 - 7.00 (m, 5H), 6.89 (d, J = 8.0 Hz, 2H), 4.86 - 4.84 (m, 2H), 3.62 - 3.58 (m, 2H), 2.30 (s, 6H). HRMS (ESI) m / z: [M + H] + calcd for C 36 H 30 IN4 + 645.1510, found: 645.1516.

[0108] Example 7:

[0109] (E)-2-(1-(2-Bromoethyl)-2-(4-(di-p-toluidino)styryl)quinolin-4(1H)-ylidene)malononitrile

[0110] QM26

[0111]

[0112] Add QM24 (1 mmol, 0.69 g), lithium bromide (1.5 mmol, 0.13 g) and 10 mL of tetrahydrofuran into a 50 mL pear-shaped flask; stir at room temperature for 12 h and monitor by TLC; after the reaction is completed, remove the solvent by distillation under reduced pressure, filter by suction and wash the solid with water to obtain 0.40 g of the target compound. Dark red solid, yield 68%. 11H NMR (400 MHz, DMSO-d6) δ 8.91 (d, J = 8.4 Hz, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.92 (t, J = 8.0 Hz, 1H), 7.66 - 7.60 (m, 3H), 7.37 (q, J = 15.6 Hz, 2H), 7.17 (d, J = 8.2 Hz, 4H), 7.00 (d, J = 8.6 Hz, 5H), 6.88 (d, J = 8.6 Hz, 2H), 4.96 (t, J = 6.8 Hz, 2H), 3.89 (t, J = 6.8 Hz, 2H), 2.29 (s, 6H). 13 13C NMR (100 MHz, DMSO-d6) δ 152.41, 150.29, 149.31, 143.91, 139.74, 137.94, 133.77, 133.46, 130.26, 129.43, 127.62, 125.24, 125.17, 120.09, 118.23, 117.98, 106.56, 99.53, 95.24, 48.49, 47.91, 42.10, 20.47. HRMS (ESI) m / z: [M + H] + calcd for C 36 H 30 BrN4 + 597.1648, found: 597.1653.

[0113] Example 8:

[0114] (E)-2-(1-(2-Azidoethyl)-2-(4-(di-p-tolylamino)styryl)quinolin-4(1H)-ylidene)malononitrile

[0115] QM27

[0116]

[0117] QM24 (5 mmol, 3.44 g), sodium azide (5.5 mmol, 0.36 g) and 25 mL of N,N-dimethylformamide were added to a 50 mL pear-shaped flask; stirred at room temperature for 12 h and monitored by TLC; after the reaction was completed, the reaction system was poured into 500 mL of ice water, filtered by suction and dried to obtain 2.24 g of the target compound. Dark red solid, yield 80%. 11H NMR (400 MHz, DMSO-d6) δ 8.91 (d, J = 8.4 Hz, 1H), 8.12 (d, J = 8.8 Hz, 1H), 7.92 (t, J = 7.6 Hz, 1H), 7.66 - 7.60 (m, 3H), 7.43 (d, J = 15.6 Hz, 1H), 7.34 (d, J = 15.6 Hz, 1H), 7.17 (d, J = 8.0 Hz, 4H), 7.06 - 6.95 (m, 5H), 6.88 (d, J = 8.4 Hz, 2H), 4.83 - 4.78 (m, 2H), 3.88 - 3.82 (m, 2H), 2.29 (s, 6H). 13 13C NMR (100 MHz, DMSO-d6) δ 152.35, 150.36, 149.25, 143.91, 139.40, 138.19, 133.61, 133.44, 130.25, 129.42, 127.66, 125.21, 125.13, 120.56, 120.06, 118.38, 118.06, 106.63, 48.89, 47.54, 46.94, 20.47. HRMS (ESI) m / z: [M+H] + calcd for C 36 H 30 N7 + 560.2557, found: 560.2559.

[0118] Example 9:

[0119] (E)-2-(1-(2-Aminoethyl)-2-(4-(di-p-tolylamino)styryl)quinolin-4(1H)-ylidene)malononitrile

[0120] QM28

[0121]

[0122] Add QM27 (5 mmol, 2.80 g), triphenylphosphine (6 mmol, 1.57 g) and 50 mL of dichloromethane into a 100 mL pear-shaped flask; stir at room temperature for 12 h and monitor by TLC; after the reaction is completed, remove the solvent by distillation under reduced pressure, add 25 mL of tetrahydrofuran and 25 mL of water. Stir and react at 75 °C in an oil bath for 12 h and monitor by TLC; after the reaction is completed, remove the solvent by distillation under reduced pressure, and perform column chromatography purification to obtain 1.25 g of the target compound. Dark red solid, with a yield of 47%. 11H NMR (400 MHz, DMSO-d6) δ 8.93 (d, J = 8.4 Hz, 1H), 8.13 (d, J = 9.0 Hz, 1H), 7.92 (t, J = 7.6 Hz, 1H), 7.67 (d, J = 8.8 Hz, 2H), 7.65 - 7.58 (m, 1H), 7.46 (d, J = 15.6 Hz, 1H), 7.35 - 7.31 (m, 1H), 7.18 (d, J = 8.2 Hz, 3H), 7.12 (d, J = 8.2 Hz, 1H), 7.02 (d, J = 7.0 Hz, 3H), 6.99 (s, 1H), 6.92 - 6.86 (m, 3H), 4.49 (t, J = 7.0 Hz, 2H), 2.99 (t, J = 7.0 Hz, 2H), 2.31 (s, 6H), 2.29 - 2.25 (m, 2H). 13 13C NMR (100 MHz, DMSO-d6) δ 153.48, 150.71, 149.71, 145.38, 144.41, 139.86, 139.35, 138.75, 133.91, 132.46, 130.73, 130.48, 128.09, 125.67, 124.45, 122.82, 120.59, 109.52, 59.87, 46.95, 46.78, 20.86. HRMS (ESI) m / z: [M+H] + calcd for C 36 H 32 N5 + 534.2652, found: 534.2648.

[0123] Example 10:

[0124] (E)-(4 - ((2-(4-(Di(p - toluidino)styryl)-4-(dicyanomethyl)quinolin - 1(4H)-yl)ethyl)amino)-4 - oxobutyl)triphenylphosphonium

[0125] QM29

[0126]

[0127] To 4-(Triphenyl - λ 4(1.2 mmol, 0.51 g) of (phosphoryl)butyric acid, EDCI (1.2 mmol, 0.23 g), 4-dimethylaminopyridine (0.2 mmol, 0.024 g) and 10 mL of N,N-dimethylformamide were added to a 50 mL pear-shaped flask; stirred at room temperature for 30 min and monitored by TLC; QM28 (1.0 mmol, 0.53 g) was added and the reaction was continued at room temperature for 12 h and monitored by TLC; after the reaction was completed, the solvent was removed by distillation under reduced pressure and purified by column chromatography to obtain 0.39 g of the target compound. Dark red solid, yield 42%. 1 HNMR (500 MHz, DMSO-d6) δ 8.89 (d, J = 8.4 Hz, 1H), 8.30 (t, J = 5.6 Hz, 1H), 8.14 (d, J = 9.0 Hz, 1H), 7.94 - 7.82 (m, 4H), 7.81 - 7.68 (m, 12H), 7.65 - 7.53 (m, 3H), 7.32 (q, 15.6 Hz, 2H), 7.17 (d, J = 8.2 Hz, 4H), 7.02 - 6.93 (m, 5H), 6.80 (d, J = 8.6 Hz, 2H), 4.58 - 4.53 (m, 2H), 3.72 - 3.42 (m, 4H), 2.30 (s, 6H), 2.29 - 2.22 (m, 2H), 1.64 (dd, J = 14.8, 7.2 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 171.76, 152.18, 150.03, 149.19, 143.84, 139.51, 138.36, 134.93, 133.53, 133.43, 130.27, 130.15, 129.38, 127.53, 125.23, 120.64, 119.90, 118.75, 118.10, 117.89, 117.48, 106.23, 46.80, 37.00, 34.99, 34.81, 20.48, 20.15, 18.03. HRMS (ESI) m / z: [M] + calcd for C 58 H 51 N5OP + 864.3826, found: 864.3830.

[0128] Example 11:

[0129] (E)-1-(4-((2-(4-(Di(p-tolyl)amino)phenyl)vinyl)-4-(dicyanomethyl)quinolin-1(4H)-yl)ethyl)amino)-4-oxobutyl)pyridin-1-ium

[0130] QM30

[0131]

[0132] 1-(3-Carboxypropyl)pyridin-1-ium (1.2 mmol, 0.29 g), HATU (1.2 mmol, 0.46 g), DIPEA (1.5 mmol, 0.19 g) and 10 mL of N,N-dimethylformamide were added to a 50 mL pear-shaped flask; stirred at room temperature for 30 min and monitored by TLC; QM28 (1.0 mmol, 0.53 g) was added and the reaction was continued at room temperature for 12 h and monitored by TLC; after the reaction was completed, the solvent was removed by distillation under reduced pressure and purified by column chromatography to obtain 0.29 g of the target compound. Dark red solid, yield 39%. 1 H NMR (500 MHz, DMSO-d6) δ 8.92 (d, J = 8.4 Hz, 1H), 8.69 (d, J = 4.0 Hz, 1H), 8.47 (d, J = 8.4 Hz, 2H), 8.22 (d, J = 9.0 Hz, 1H), 7.97 (t, J = 8.0 Hz, 1H), 7.91 (t, J = 5.4 Hz, 1H), 7.70 (d, J = 8.8 Hz, 2H), 7.68 - 7.61 (m, 1H), 7.46 (dd, J = 8.4, 4.4 Hz, 1H), 7.39 (s, 2H), 7.19 (d, J = 8.2 Hz, 4H), 7.04 (s, 1H), 6.99 (d, J = 8.4 Hz, 4H), 6.89 (d, J = 8.8 Hz, 2H), 4.84 - 4.72 (m, 2H), 3.65 - 3.62 (m, 4H), 3.58 - 3.49 (m, 2H), 3.17 - 3.14 (m, 2H), 2.30 (s, 6H). HRMS (ESI) m / z: [M] + calcd for C 45 H 41 N6O + 681.3336, found: 681.3340.

[0133] Example 12:

[0134] (E)-2-(2-(4-(Diphenylamino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile

[0135] QM09

[0136]

[0137] The synthesis method was the same as that of Example 4, except that 4-(di-p-toluidino)benzaldehyde was replaced by 4-(diphenylamino)benzaldehyde. Dark red solid, yield 72%. 11H NMR (400 MHz, DMSO-d6) δ 8.92 (dd, J = 8.4, 1.2 Hz, 1H), 8.09 (d, J = 9.0 Hz, 1H), 7.97 - 7.86 (m, 1H), 7.70 - 7.50 (m, 4H), 7.39 - 7.29 (m, 5H), 7.16 - 7.07 (m, 6H), 7.03 (s, 1H), 6.97 (d, J = 8.8 Hz, 2H), 5.18 (t, J = 5.6 Hz, 1H), 4.63 (t, J = 5.2 Hz, 2H), 3.85 (c, 2H). 13 13C NMR (100 MHz, DMSO-d6) δ 152.21, 150.66, 148.77, 146.49, 138.85, 138.47, 133.39, 129.76, 129.34, 128.74, 125.03, 124.88, 124.05, 121.59, 120.68, 119.54, 119.38, 118.56, 106.64, 58.91, 50.50, 46.67. HRMS (ESI) m / z: [M+H] + calcd for C 34 H 27 N4O + 507.2179, found: 507.2171.

[0138] Example 13:

[0139] (E)-2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile

[0140] QM11

[0141]

[0142] The synthesis method is the same as that of Example 4, except that 4-(di-p-toluidino)benzaldehyde is replaced by 4-(bis(4-methoxyphenyl)amino)benzaldehyde. Dark red solid, with a yield of 76%. 11H NMR (400 MHz, DMSO-d6) δ 8.90 (d, J = 8.4 Hz, 1H), 8.07 (d, J = 9.0 Hz, 1H), 7.88 (t, J = 7.8 Hz, 1H), 7.62 - 7.53 (m, 3H), 7.45 (d, J = 15.6 Hz, 1H), 7.27 (d, J = 15.6 Hz, 1H), 7.09 (d, J = 8.8 Hz, 4H), 7.01 (s, 1H), 6.95 (d, J = 8.8 Hz, 4H), 6.75 (d, J = 8.6 Hz, 2H), 5.17 (t, J = 5.6 Hz, 1H), 4.64 - 4.58 (m, 2H), 3.88 - 3.822 (m, 2H), 3.76 (s, 6H). 13 13C NMR (100 MHz, DMSO-d6) δ 156.39, 152.11, 150.86, 149.95, 139.26, 139.20, 138.53, 133.37, 129.31, 127.40, 126.50, 125.03, 124.87, 120.70, 118.58, 118.04, 117.93, 115.12, 106.44, 58.93, 55.31, 50.44, 46.28. HRMS (ESI) m / z: [M+H] + calcd for C 36 H 31 N4O3 + 567.2391, found: 567.2388.

[0143] Example 14:

[0144] (E)-2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile

[0145] QM12

[0146]

[0147] The synthesis method was the same as that of Example 4, except that 4-(di-p-toluidino)benzaldehyde was replaced with 4-(bis(4-fluorophenyl)amino)benzaldehyde. Dark red solid, with a yield of 58%. 11H NMR (400 MHz, DMSO-d6) δ 8.91 (dd, J = 8.4, 1.2 Hz, 1H), 8.08 (d, J = 8.8 Hz, 1H), 7.89 (ddd, J = 8.8, 7.2, 1.2 Hz, 1H), 7.68 - 7.57 (m, 3H), 7.53 (d, J = 15.8 Hz, 1H), 7.32 (d, J = 15.8 Hz, 1H), 7.26 - 7.12 (m, 8H), 7.02 (s, 1H), 6.89 (d, J = 8.8 Hz, 2H), 5.18 (t, J = 5.6 Hz, 1H), 4.63 (t, J = 5.2 Hz, 2H), 3.85 (q, J = 5.4 Hz, 2H). 13 13C NMR (100 MHz, DMSO-d6) δ 159.34 (d, J = 241.9 Hz), 152.65, 151.09, 149.42, 143.21, 139.30, 138.93, 133.84, 129.85, 127.67 (d, J = 8.3 Hz), 125.49, 125.34, 121.13, 120.75, 119.82, 119.00, 117.10 (d, J = 22.6 Hz), 107.06, 59.38, 50.96, 47.13. HRMS (ESI) m / z: [M+H] + calcd for C 34 H 25 F2N4O + 543.1991, found: 543.1998.

[0148] Example 15:

[0149] (E)-2-(2-(2-(5-(4-(Diphenylamino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile

[0150] QM13

[0151]

[0152] The synthesis method was the same as that of Example 4, except that 4-(di-p-tolylamino)benzaldehyde was replaced with 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde. Dark red solid, yield 67%. 11H NMR (400 MHz, DMSO-d6) δ 9.02 - 8.82 (m, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.94 - 7.86 (m, 1H), 7.64 - 7.56 (m, 4H), 7.53 (d, J = 3.8 Hz, 1H), 7.46 (d, J = 3.8 Hz, 1H), 7.42 - 7.32 (m, 5H), 7.13 - 7.06 (m, 6H), 7.03 (s, 1H), 6.99 (d, J = 8.8 Hz, 2H), 5.22 (t, J = 5.6 Hz, 1H), 4.61 (t, J = 4.8 Hz, 2H), 3.88 (q, J = 5.4 Hz, 2H), 3.30 (s, 3H). 13 13C NMR (100 MHz, DMSO-d6) δ 152.14, 149.91, 147.52, 146.66, 145.82, 138.62, 138.39, 133.43, 132.76, 132.29, 129.70, 126.67, 125.06, 124.88, 124.61, 123.73, 122.47, 120.66, 119.68, 119.31, 118.43, 106.57, 58.93, 56.03, 50.68, 46.91, 18.56. HRMS (ESI) m / z: [M + H] + calcd for C 38 H 29 N4OS + 589.2057, found: 589.2063.

[0153] Example 16:

[0154] (E)-2-(2-(2-(5-(4-(Di(p-tolyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)propanedinitrile

[0155] QM14

[0156]

[0157] The synthesis method was the same as that of Example 4, except that 4-(di(p-tolyl)amino)benzaldehyde was replaced with 5-(4-(di(p-tolyl)phenyl)thiophen-2-carbaldehyde). Dark red solid, yield 73%. 11H NMR (400 MHz, DMSO-d6) δ 8.92 (dd, J = 8.4, 1.2 Hz, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.94 - 7.86 (m, 1H), 7.63 - 7.54 (m, 4H), 7.51 (d, J = 3.8 Hz, 1H), 7.42 (d, J = 3.8 Hz, 1H), 7.37 (d, J = 15.6 Hz, 1H), 7.15 (d, J = 8.2 Hz, 4H), 7.02 (s, 1H), 6.97 (d, J = 8.4 Hz, 4H), 6.90 (d, J = 8.8 Hz, 2H), 5.22 (t, J = 5.4 Hz, 1H), 4.60 (t, J = 4.7 Hz, 2H), 3.88 (q, J = 5.4 Hz, 2H), 2.28 (s, 6H). 13 13C NMR (100 MHz, DMSO-d6) δ 152.63, 150.50, 148.44, 146.55, 144.63, 138.89, 138.83, 133.94, 133.53, 130.69, 127.04, 126.13, 125.43, 123.92, 121.54, 121.15, 120.01, 118.98, 118.95, 107.04, 59.42, 51.14, 47.28, 20.92. HRMS (ESI) m / z: [M + H] + calcd for C 40 H 33 N4OS + 617.2370, found: 617.2378.

[0158] Example 17:

[0159] (E)-2-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)propanedinitrile

[0160] QM15

[0161]

[0162] The synthesis method was the same as that of Example 4, except that 44-(di-p-toluidino)benzaldehyde was replaced with 5-(4-(bis(4-methoxyphenyl)amino)phenyl)thiophene-2-carbaldehyde. Dark red solid, yield 60%. 11H NMR (400 MHz, DMSO-d6) δ 8.90 (d, J = 8.4 Hz, 1H), 8.04 (d, J = 8.8 Hz, 1H), 7.88 (t, J = 7.8 Hz, 1H), 7.57 (dd, J = 15.6, 8.4 Hz, 2H), 7.51 (d, J = 8.8 Hz, 2H), 7.48 (d, J = 4.0 Hz, 1H), 7.38 - 7.30 (m, 2H), 7.07 (d, J = 8.8 Hz, 4H), 7.00 (s, 1H), 6.94 (d, J = 8.8 Hz, 4H), 6.77 (d, J = 8.8 Hz, 2H), 5.25 (t, J = 5.4 Hz, 1H), 4.62 - 4.56 (m, 2H), 3.88 (q, J = 5.4 Hz, 2H), 3.75 (s, 6H). 13 13C NMR (100 MHz, DMSO-d6) δ 156.65, 152.60, 150.51, 149.20, 146.91, 139.93, 138.90, 138.46, 133.91, 133.41, 132.92, 127.66, 127.00, 125.55, 125.38, 124.82, 123.49, 121.15, 119.83, 119.73, 119.16, 118.94, 115.55, 107.02, 59.40, 55.76, 51.14, 47.24. HRMS (ESI) m / z: [M+H] + calcd for C 40 H 33 N4O3S + 649.2268, found: 649.2274.

[0163] Example 18:

[0164] (E)-2-(2-(2-(5-(4-(Bis(4-fluorophenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile

[0165] QM16

[0166]

[0167] The synthesis method was the same as that of Example 4, except that 4-(di-p-toluidino)benzaldehyde was replaced with 5-(4-(bis(4-fluorophenyl)amino)phenyl)thiophene-2-carbaldehyde. Dark red solid, yield 44%. 11H NMR (400 MHz, DMSO-d6) δ 8.90 (d, J = 8.2 Hz, 1H), 8.05 (d, J = 8.4 Hz, 1H), 7.88 (t, J = 7.4 Hz, 1H), 7.64 - 7.46 (m, 5H), 7.46 - 7.30 (m, 2H), 7.24 - 7.10 (m, 8H), 7.00 (s, 1H), 6.90 (d, J = 8.2 Hz, 2H), 5.30 - 5.22 (m, 1H), 4.68 - 4.56 (m, 2H), 3.95 - 3.85 (m, 2H). 13 13C NMR (100 MHz, DMSO-d6) δ 159.18 (d, J = 241.6 Hz), 152.66, 150.49, 148.23, 146.31, 143.42 (d, J = 2.3 Hz), 139.05, 138.90, 133.95, 133.30, 132.82, 127.41 (d, J = 8.3 Hz), 127.22, 126.74, 125.56, 125.42, 124.19, 121.66, 121.16, 120.20, 118.98, 117.06 (d, J = 22.7 Hz), 107.08, 59.40, 51.17, 47.31. HRMS (ESI) m / z: [M+H] + calcd for C 38 H 27 F2N4OS + 625.1868, found: 625.1875.

[0168] Example 19:

[0169] (E)-2-(2-(4-(Diphenylamino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile

[0170] QM02

[0171]

[0172] The synthesis method was the same as that of Example 2, except that 4-(di-p-toluidino)benzaldehyde was replaced by 4-(diphenylamino)benzaldehyde. Red solid, with a yield of 75%. 11H NMR (400 MHz, DMSO-d6) δ 8.96 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 9.2 Hz, 1H), 7.96 (t, J = 7.6 Hz, 1H), 7.73 (d, J = 8.4 Hz, 2H), 7.66 (t, J = 7.4 Hz, 1H), 7.50 (d, J = 15.6 Hz, 1H), 7.43 - 7.37 (m, 5H), 7.20 - 7.12 (m, 6H), 7.05 (s, 1H), 6.99 (d, J = 8.4 Hz, 2H), 5.01 - 4.54 (m, 2H), 2.99 (t, J = 7.0 Hz, 2H), 2.15 (s, 3H). HRMS (ESI) m / z: [M+H] + calcd for C 35 H 29 N4S + : 537.2107, found: 537.2102.

[0173] Example 20:

[0174] (E)-2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile

[0175] QM04

[0176]

[0177] The synthesis method was the same as that of Example 2, except that 4-(di-p-toluidino)benzaldehyde was replaced by 4-(bis(4-methoxyphenyl)amino)benzaldehyde. Red solid, with a yield of 71%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.90 (d, J = 8.0 Hz, 1H), 8.05 (d, J = 8.6 Hz, 1H), 7.91 (t, J = 7.2 Hz, 1H), 7.59 (d, J = 7.8 Hz, 3H), 7.34 (d, J = 6.6 Hz, 2H), 7.11 (d, J = 8.2 Hz, 4H), 7.01 - 6.88 (m, 5H), 6.73 (d, J = 8.0 Hz, 2H), 4.75 - 4.67 (m, 2H), 3.76 (s, 6H), 2.98 - 2.90 (m, 2H), 2.12 (s, 3H). HRMS (ESI) m / z: [M+H] + calcd for C 37 H 33 N4O2S + 597.2319, found: 597.2325.

[0178] Example 21:

[0179] (E)-2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile

[0180] QM05

[0181]

[0182] The synthesis method is the same as that of Example 2, except that 4-(di-p-toluidino)benzaldehyde is replaced by 4-(bis(4-fluorophenyl)amino)benzaldehyde. Red solid, with a yield of 67%. 1 H NMR(400MHz,DMSO-d6)δ8.92(d,J=8.0Hz,1H),8.06(d,J=9.0Hz,1H),7.92(t,J=8.0Hz,1H),7.68(d,J=8.8Hz,2H),7.64-7.58(m,1H),7.45(d,J=15.6Hz,1H),7.35(d,J=15.6Hz,1H),7.22(t,J=8.8Hz,4H),7.18-7.12(m,4H),7.00(s,1H),6.87(d,J=8.6Hz,2H),4.72(t,J=7.2Hz,2H),2.95(t,J=7.2Hz,2H),2.11(s,3H).HRMS(ESI)m / z:[M+H] + calcd for C 35 H 27 F2N4S + 573.1919,found:573.1909.

[0183] Example 22:

[0184] (E)-2-(2-(2-(5-(4-(Diphenylamino)phenyl)thiophen-2-yl)vinyl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile

[0185] QM06

[0186]

[0187] The synthesis method is the same as that of Example 2, except that 4-(di-p-toluidino)benzaldehyde is replaced by 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde. Dark red solid, with a yield of 67%. 11H NMR (400 MHz, DMSO-d6) δ 8.92 (d, J = 8.8 Hz, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.95 - 7.90 (m, 1H), 7.65 - 7.62 (m, 2H), 7.60 - 7.55 (m, 3H), 7.40 - 7.32 (m, 3H), 7.00 (s, 1H), 6.94 (d, J = 8.6 Hz, 3H), 7.20 - 7.16 (m, 1H), 7.09 (d, J = 7.2 Hz, 2H), 7.06 - 7.02 (m, 5H), 4.84 (t, J = 6.8 Hz, 2H), 2.98 (t, J = 6.8 Hz, 2H), 2.17 (s, 3H). HRMS (ESI) m / z: [M+H] + calcd for C 39 H 31 N4S2 + 619.1985, found: 619.1980.

[0188] Example 23:

[0189] (E)-2-(2-(2-(5-(4-(Di-p-tolylamino)phenyl)thiophen-2-yl)vinyl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile

[0190] QM07

[0191]

[0192] The synthesis method was the same as that of Example 2, except that 4-(di-p-tolylamino)benzaldehyde was replaced with 5-(4-(di-p-tolyl)phenyl)thiophene-2-carbaldehyde. Dark red solid, yield 59%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.92 (dd, J = 8.4, 1.2 Hz, 1H), 8.07 (d, J = 8.8 Hz, 1H), 7.95 - 7.90 (m, 1H), 7.68 - 7.60 (m, 2H), 7.58 - 7.55 (m, 3H), 7.44 (d, J = 4.0 Hz, 1H), 7.24 (d, J = 15.6 Hz, 1H), 7.16 (d, J = 8.4 Hz, 4H), 7.02 (s, 1H), 6.98 (d, J = 8.4 Hz, 4H), 6.91 (d, J = 8.8 Hz, 2H), 4.82 - 4.54 (m, 2H), 3.05 - 2.85 (m, 2H), 2.29 (s, 6H), 2.17 (s, 3H). HRMS (ESI) m / z: [M+H] + calcd for C 41 H35 N4S2 + 647.2298, found: 647.2231.

[0193] Example 24:

[0194] (E)-2-(2-(2-(5-(4-(bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile

[0195] QM08

[0196]

[0197] The synthesis method is the same as that of Example 2, except that 4-(di-p-toluidino)benzaldehyde is replaced by 5-(4-(bis(4-methoxyphenyl)amino)phenyl)thiophene-2-carboxaldehyde. Dark red solid, with a yield of 52%. 1 H NMR (400 MHz, DMSO-d6) δ 8.90 (d, J = 8.2 Hz, 1H), 8.05 (d, J = 8.8 Hz, 1H), 7.91 (t, J = 8.0 Hz, 1H), 7.65 - 7.54 (m, 2H), 7.53 - 7.45 (m, 3H), 7.39 - 7.37 (m, 1H), 7.19 (d, J = 15.6 Hz, 1H), 7.08 (d, J = 8.6 Hz, 4H), 7.00 (s, 1H), 6.94 (d, J = 8.6 Hz, 3H), 6.77 (d, J = 8.4 Hz, 2H), 4.87 - 4.30 (m, 2H), 3.75 (s, 6H), 2.96 (t, J = 6.8 Hz, 2H), 2.17 (s, 3H). HRMS (ESI) m / z: [M+H] + calcd for C 41 H 35 N4O2S2 + 679.2196, found: 679.2190.

[0198] Example 25:

[0199] (E)-(2-(4-(diphenylamino)-2-4-(dicyanoimino)quinolin-1(4H)-yl)ethyl)dimethyl sulfonate

[0200] QM17

[0201]

[0202] The synthesis method is the same as that of Example 3, except that QM03 is replaced by QM02. Dark red solid, with a yield of 22%. 11H NMR (400 MHz, DMSO-d6) δ 8.94 (d, J = 8.0 Hz, 1H), 8.10 (d, J = 8.0 Hz, 1H), 8.06 - 7.91 (m, 2H), 7.74 (d, J = 8.0 Hz, 2H), 7.68 - 7.61 (m, 2H), 7.43 - 7.33 (m, 6H), 7.16 - 7.73 (m, 2H), 7.10 (d, J = 7.2 Hz, 3H), 7.04 (s, 1H), 6.96 (d, J = 7.4 Hz, 2H), 5.12 - 5.04 (m, 2H), 3.85 - 3.76 (m, 2H), 2.96 (s, 6H). 13 13C NMR (100 MHz, DMSO-d6) δ 152.66, 149.76, 149.08, 146.46, 146.40, 134.00, 129.84, 129.80, 129.76, 129.43, 128.71, 128.32, 125.34, 125.28, 125.09, 125.02, 124.29, 121.16, 120.62, 106.87, 48.54, 42.44, 25.18. HRMS (ESI) m / z: [M] + calcd for C 36 H 31 N4S + 551.2264, found: 551.2260.

[0203] Example 26:

[0204] (E)-(2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate

[0205] QM19

[0206]

[0207] The synthesis method is the same as that of Example 3, except that QM03 is replaced with QM04. Dark red solid, with a yield of 28%. 11H NMR (400 MHz, DMSO-d6) δ 8.95 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 8.8 Hz, 1H), 7.98 (t, J = 8.0 Hz, 1H), 7.67 (t, J = 9.0 Hz, 3H), 7.41 (d, J = 15.6 Hz, 1H), 7.32 (d, J = 15.6 Hz, 1H), 7.12 (t, J = 6.0 Hz, 4H), 7.05 (s, 1H), 6.99 (d, J = 8.8 Hz, 4H), 6.77 (d, J = 8.8 Hz, 2H), 5.07 (t, J = 6.8 Hz, 2H), 3.80 - 3.75 (m, 8H), 2.96 (s, 6H). 13 13C NMR (101 MHz, DMSO) δ 156.56, 152.55, 150.27, 149.92, 140.75, 139.03, 138.03, 133.93, 129.74, 127.59, 126.09, 125.36, 125.28, 120.66, 118.98, 117.96, 117.55, 116.20, 115.19, 106.60, 79.00, 55.35, 48.21, 42.33, 25.19.

[0208] HRMS (ESI) m / z: [M] + calcd for C 38 H 35 N4O2S + 611.2475, found: 611.2479.

[0209] Example 27:

[0210] (E)-(2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate

[0211] QM20

[0212]

[0213] The synthesis method was the same as that of Example 3, except that QM03 was replaced by QM05. Dark red solid, with a yield of 19%. 11H NMR (400 MHz, DMSO-d6) δ 8.96 (d, J = 8.4 Hz, 1H), 8.11 (d, J = 9.0 Hz, 1H), 8.04 - 7.96 (m, 1H), 7.73 (d, J = 8.6 Hz, 2H), 7.68 (d, J = 7.6 Hz, 1H), 7.42 (d, J = 6.4 Hz, 2H), 7.30 - 7.21 (m, 4H), 7.20 - 7.15 (m, 4H), 7.06 (s, 1H), 6.91 (d, J = 8.6 Hz, 2H), 5.07 (t, J = 6.8 Hz, 2H), 3.78 (t, J = 6.8 Hz, 2H), 2.96 (s, 6H). HRMS (ESI) m / z: [M] + calcd for C 36 H 29 F2N4S + 589.2076, found: 589.2070.

[0214] Example 28:

[0215] (E)-(2-(4-(Dicyanomethylene)-2-(2-(5-(4-(Diphenylamino)phenyl)thiophen-2-yl)vinyl)quinolin-1(4H)-yl)ethyl)dimethylsulfonate

[0216] QM21

[0217]

[0218] The synthesis method was the same as that of Example 3, except that QM03 was replaced by QM06. Dark red solid, with a yield of 17%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.94 (d, J = 8.8 Hz, 1H), 8.11 (d, J = 8.8 Hz, 1H), 8.02 - 7.94 (m, 1H), 7.65 - 7.58 (m, 4H), 7.52 - 7.49 (m, 1H), 7.39 - 7.33 (m, 5H), 7.23 - 7.18 (m, 1H), 7.13 (d, J = 7.2 Hz, 2H), 7.10 - 7.07 (m, 5H), 7.05 (s, 1H), 7.01 (d, J = 8.6 Hz, 3H), 5.05 (t, J = 6.8 Hz, 2H), 3.78 (t, J = 6.8 Hz, 2H), 2.99 (s, 6H). 1313C NMR (100 MHz, DMSO-d6) δ 152.61, 148.93, 146.62, 138.33, 138.01, 133.78, 133.38, 129.75, 126.71, 125.34, 124.68, 124.60, 124.54, 123.85, 123.76, 122.57, 122.43, 120.56, 118.77, 117.97, 117.86, 48.79, 25.14, 21.14. HRMS (ESI) m / z: [M] + calcd for C 40 H 33 N4S2 + 633.2141, found: 633.2141.

[0219] Example 29:

[0220] (E)-(2-(2-(5-(4-(Di-p-tolylamino)phenyl)thiophen-2-yl)vinyl)-4-(dicyanomethylene)quinolin-1(4H)-yl)ethyl)dimethylsulfonate

[0221] QM22

[0222]

[0223] The synthesis method is the same as that of Example 3, except that QM03 is replaced by QM07. Dark red solid, with a yield of 26%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.93 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 8.8 Hz, 1H), 7.97 (t, J = 7.8 Hz, 1H), 7.73 - 7.63 (m, 2H), 7.60 (d, J = 3.8 Hz, 1H), 7.56 (d, J = 8.6 Hz, 2H), 7.46 (d, J = 3.8 Hz, 1H), 7.22 - 7.13 (m, 5H), 7.04 (s, 1H), 6.98 (d, J = 8.2 Hz, 4H), 6.91 (d, J = 8.8 Hz, 2H), 5.04 (t, J = 6.8 Hz, 2H), 3.77 (t, J = 6.8 Hz, 2H), 2.98 (s, 6H), 2.28 (s, 6H). 1313C NMR (100 MHz, DMSO-d6) δ 152.58, 148.96, 148.11, 146.78, 144.10, 138.04, 136.62, 133.99, 133.84, 133.45, 133.18, 130.24, 128.45, 126.58, 125.54, 125.34, 125.01, 123.54, 121.00, 120.57, 118.79, 117.96, 117.65, 106.85, 48.74, 42.41, 25.15, 20.45. HRMS (ESI) m / z: [M] + calcd for C 42 H 37 N4S2 + 661.2454, found: 661.2450.

[0224] Example 30:

[0225] (E)-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-4-(dicyanomethylene)quinolin-1(4H)-yl)ethyl)dimethylsulfonate

[0226] QM23

[0227]

[0228] The synthesis method is the same as that of Example 3, except that QM03 is replaced by QM08. Dark red solid, with a yield of 24%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.94 (d, J = 8.4 Hz, 1H), 8.11 (d, J = 8.6 Hz, 1H), 7.97 (t, J = 7.6 Hz, 1H), 7.74 - 7.63 (m, 2H), 7.59 (d, J = 2.6 Hz, 1H), 7.52 (d, J = 8.0 Hz, 2H), 7.43 (d, J = 2.6 Hz, 1H), 7.16 (d, J = 15.5 Hz, 1H), 7.09 (d, J = 8.4 Hz, 4H), 7.04 (s, 1H), 6.95 (d, J = 8.2 Hz, 4H), 6.79 (d, J = 8.0 Hz, 2H), 5.08 - 5.00 (m, 2H), 3.81 - 3.72 (m, 8H), 2.98 (s, 6H). 1313C NMR(100MHz, DMSO-d6) δ 156.24, 152.56, 148.99, 148.86, 147.13, 139.37, 138.02, 137.69, 133.98, 133.89, 133.51, 127.25, 126.54, 125.33, 124.21, 123.12, 120.57, 118.61, 117.39, 115.11, 106.80, 55.28, 48.67, 36.72, 34.60, 25.15. HRMS(ESI) m / z: [M] + calcd for C 42 H 37 N4O2S2 + 693.2352, found: 693.2359.

[0229] In addition to the compounds in the above examples, considering that the anti-tumor effect of photosensitizers is closely related to their optical properties, the present invention characterized the absorption wavelength, emission wavelength, molar extinction coefficient and AIE properties of some compounds in the examples and evaluated their anti-tumor activities at the cellular level.

[0230] Example 31: Determination of the Absorption Wavelength, Emission Wavelength and Molar Extinction Coefficient of Compounds

[0231] Experimental Purpose: To characterize the characteristic absorption wavelength, emission wavelength and molar extinction coefficient of compounds.

[0232] Experimental Method: Determined by ultraviolet-visible spectrophotometer and fluorescence spectrophotometer. Absorption spectrum determination: The compound was configured into a DMSO solution with a concentration of 20 μM, and the absorption curve was measured in the range of 200 - 800 nm using an ultraviolet-visible spectrophotometer, and the corresponding molar extinction coefficient was calculated according to the absorbance. Fluorescence spectrum determination: The compound was configured into a DMSO solution with a concentration of 20 μM, and the fluorescence emission curve was measured at 400 - 800 nm using a fluorescence spectrophotometer.

[0233] Experimental Results: See Table 1 and Appendix Figure 1 .

[0234] Table 1 Photophysical Properties of Some Compounds

[0235]

[0236]

[0237] Example 32: Determination of AIE Properties

[0238] Experimental Purpose: To characterize the aggregation-induced fluorescence enhancement property of compounds.

[0239] Experimental method: Determined by fluorescence spectrophotometer. Prepare DMSO / H2O solutions with different ratios (water content is set to 0%, 20%, 40%, 60%, 80%, 99%), add the compound to make the final concentration of 20 μM, and use a fluorescence spectrophotometer to measure the fluorescence emission curve at 400-800 nm.

[0240] Experimental results: see attached Figure 2 , 3. It can be observed that both QM10 and QM18 have the strongest fluorescence emission peak in 20% DMSO / 80% H2O solution, and solid QM10 and QM18 also show visible red fluorescence under 365nm ultraviolet light excitation, such as Figure 2 and Figure 3 As shown in (c) and (d) in Fig. Figure 2 and Figure 3 As shown in (a) and (b), the change curves of the fluorescence intensity under different water contents present a "V" shape. The reason is the combined influence of the ICT and AIE effects. Since water has a large polarity, initially with the increase of water content, the ICT effect causes the fluorescence intensity to decrease; and with the further increase of water content, the AIE effect gradually increases, causing the fluorescence intensity to increase.

[0241] Example 33: Evaluation of the reactive oxygen species generating ability of photosensitizer molecules

[0242] Experimental purpose: To characterize the total reactive oxygen species generating capacity of photosensitizers.

[0243] Experimental method: Determined by fluorescence spectrophotometer. The compound (concentration of 10 μM) was added to an aqueous solution containing the active oxygen indicator DCFH (concentration of 50 μM). 2 ) After irradiation for 30 seconds, the fluorescence spectrum of the solution in the range of 500-650 nm (excitation wavelength is 480 nm) was measured. The fluorescence intensity at 525 nm was recorded to indicate the total active oxygen generation rate.

[0244] Experimental results: see attached Figure 4 According to Figure 4 It can be seen that after adding the photosensitizer, the amount of active oxygen generated increases as the illumination time increases.

[0245] Example 34: Evaluation of the singlet oxygen generating ability of photosensitizer molecules

[0246] Experimental purpose: To characterize the singlet oxygen generating ability of photosensitizers.

[0247] Experimental method: Determined by ultraviolet-visible spectrophotometer. The compound (at a concentration of 10 μM) was added to an aqueous solution containing the singlet oxygen indicator ABDA (at a concentration of 50 μM). After irradiating with white light (400 - 800 nm, 20 mW / cm 2 ) for 30 s each time, the ultraviolet-visible absorption spectrum of the solution in the range of 300 - 650 nm was measured. The absorbance at 400 nm was recorded to represent the singlet oxygen generation rate.

[0248] Experimental results: See the appendix Figure 5 . According to as Figure 5 it can be seen that after adding the photosensitizer, with the extension of the irradiation time, the amount of singlet oxygen generated increases. Among them, the compounds with hydroxyl side chains do not have the ability to generate singlet oxygen.

[0249] Example 35: Evaluation of the hydroxyl radical generation ability of photosensitizer molecules

[0250] Experimental purpose: To characterize the hydroxyl radical generation ability of the photosensitizer.

[0251] Experimental method: Determined by fluorescence spectrophotometer. The compound (at a concentration of 10 μM) was added to an aqueous solution containing the hydroxyl radical indicator HPF (at a concentration of 20 μM). After irradiating with white light (400 - 800 nm, 20 mW / cm 2 ) for 30 s each time, the fluorescence spectrum (excitation wavelength: 480 nm) of the solution in the range of 500 - 650 nm was measured. The fluorescence intensity at 514 nm was recorded to represent the total reactive oxygen species generation rate.

[0252] Experimental results: See the appendix Figure 6 . According to as Figure 6 it can be seen that after adding the photosensitizer, with the extension of the irradiation time, the amount of hydroxyl radicals generated increases.

[0253] Example 36: Evaluation of the organelle localization of photosensitizer molecules

[0254] Experimental purpose: To characterize the organelle targeting ability of the photosensitizer.

[0255] Experimental method: 4T1 cells were cultured in a 35-mm glass-bottom cell culture dish with RPMI 1640 medium containing 5% fetal bovine serum and 1% penicillin-streptomycin in an incubator at 37 °C with 5% CO2. When the cells proliferated to 80%, the culture medium was removed, and the cells were washed twice with PBS solution. Then, a medium containing a photosensitizer at a concentration of 5 μM was added to the 4T1 cell culture dish and cultured at 37 °C for a certain period of time. After rinsing three times with PBS solution to remove the photosensitizer that did not enter the cells, mitochondrial targeting probe, lipid droplet targeting probe, lysosome targeting probe, and endoplasmic reticulum targeting probe were added respectively. After incubating for 15 min, the culture medium was removed, and the cells were washed three times with PBS solution, then fresh culture medium was added, and the cells were observed under a confocal microscope.

[0256] Experimental results: See the appendix Figure 7 According to Figure 7 it can be seen that the fluorescence image of the photosensitizer in the cells highly coincides with the endoplasmic reticulum probe, indicating that the photosensitizer can specifically target the endoplasmic reticulum.

[0257] Example 37: MTT method for testing the photodynamic activity and dark toxicity of photosensitizers

[0258] Experimental purpose: Using the photosensitizer chlorin e6 that has entered clinical research as a positive control, the cytotoxicity of some compounds was evaluated by the thiazolyl blue MTT colorimetric method, and the cytotoxicity of the compounds to tumors under dark and light conditions was detected.

[0259] Experimental methods: (1) Cell lines and cell culture: 4T1 cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin in an incubator at 37°C with 5% CO2; Hela cells were cultured in DMEN medium containing 10% fetal bovine serum and 1% penicillin-streptomycin in an incubator at 37°C with 5% CO2. (2) Dark cytotoxicity assay of cells: 4T1 and Hela cells in good growth condition were inoculated into the corresponding 96-well plates with 200 μL of medium containing 5000 cells per well, and cultured overnight in an incubator at 37°C with 5% CO2. 1 μL of DMSO solution containing photosensitizer was added to make the final concentrations 20 - 100 μM respectively, and 3 replicate wells were set. At the same time, replicate wells with 1 μL of DMSO (without photosensitizer) were set as the control group, and cultured in the dark for 24 hours. The 96-well plates were taken out, and the medium was changed to remove the drugs that did not enter the cells (washed 1 - 2 times with PBS and fresh medium was added). 15 μL of MTT solution with a concentration of 50 mg / mL was added to each well, cultured for 4 hours, the medium was removed and 150 μL of DMSO was added to each well. Finally, the absorbance value (OD) of each well was measured at 490 nm with an enzyme-linked immunosorbent assay (ELISA) reader, and the cell survival rate was calculated. (3) Phototoxicity assay of cells: 4T1 and Hela cells in good growth condition were inoculated into the corresponding 96-well plates with 200 μL of medium containing 5000 cells per well, and cultured overnight in an incubator at 37°C with 5% CO2. 1 μL of DMSO solution containing photosensitizer was added to make the final concentrations 0.1 - 20 μM respectively, and 3 replicate wells were set. At the same time, replicate wells with 1 μL of DMSO (without photosensitizer) were set as the control group, and cultured in the dark for 24 hours. The 96-well plates were taken out, and the medium was changed to remove the drugs that did not enter the cells (washed 1 - 2 times with PBS and fresh medium was added), and irradiated with white light for 10 min (wavelength 400 - 800 nm, power 20 mW / cm 2 ). After light treatment, it was cultured in the dark for 24 hours. 15 μL of MTT solution with a concentration of 50 mg / mL was added to each well, cultured for 4 hours, the medium was removed and 150 μL of DMSO was added to each well. Finally, the absorbance value (OD) of each well was measured at 490 nm with an enzyme-linked immunosorbent assay (ELISA) reader, and the cell survival rate was calculated. For some compounds, curves were plotted using GraphPad Prism 8, and the phototoxicity IC 50 was calculated by fitting the curves.

[0260] Experimental results: See Table 2 and Appendix Figure 8 . It can be seen from the data results in Table 2 that the tested compounds all had relatively low cytotoxicity under dark conditions (IC 50Larger), while after photoexcitation, the cytotoxicity increases sharply, and some of these compounds have stronger phototoxicity than the positive control chlorin e6, indicating that this type of compound is an excellent photosensitizer capable of killing tumor cells.

[0261] Table 2 Cytotoxicity of Compounds

[0262]

[0263] Example 38: Photodynamic Therapy of Photosensitizer in Mice Bearing Breast Cancer

[0264] Experimental Purpose: To evaluate the anti-tumor effect of the photosensitizer in vivo.

[0265] Experimental Method: Select the photosensitizer QM10 for in vivo photodynamic anti-tumor activity evaluation. Implant 5*10 5 4T1 breast cancer cells subcutaneously in mice. When the tumor volume grows to about 50 mm 3 , randomly divide the experimental mice into three groups. The first and second groups of mice are given 0.6 mg / kg of the photosensitizer QM10, and the third group of mice is given an equal volume of PBS solution; about 30 min after administration, select a white light lamp (200 mW / cm 2 ) to irradiate the first group of mice and the third group of mice for 15 min. The second group of mice is placed in the dark after administration. Administer the drug and irradiate on the first and third days, and measure the changes in tumor volume and mouse body weight every other day. When the tumor volume of the experimental group of mice reaches about 600 mm 3 (after 15 days), sacrifice the mice, dissect the tumors and weigh the tumor masses. The results are as Figure 9 shown, and QM10 shows good in vivo anti-tumor activity.

[0266] In summary, the present invention is based on the Quinoline-Malononitrile (QM) core with Aggregation-Induced Emission (AIE) properties as the backbone. By introducing diverse substituent side chains at the nitrogen atom at the 1-position of the QM core and further introducing diverse triphenylamine (TPA) fragments through the methyl group at the 2-position carbon atom, a D-π-A type AIE photosensitizer molecule is constructed. The above structural modification can affect its Intramolecular Charge Transfer (ICT) effect by regulating the photophysical properties of the photosensitizer and the electron-donating and electron-withdrawing abilities of the D-π-A system, effectively reducing the energy level difference (ΔE ST), thus promoting the efficient generation of reactive oxygen species (ROS) and transforming the QM parent nucleus without photosensitizing activity into a novel AIE photosensitizer molecule. Judging from the experimental results, the maximum absorption wavelength of the target compound is within 430 nm to 474 nm, the maximum emission wavelength is within 590 nm to 663 nm, and the molar extinction coefficient is within 2600 M -1 CM -1 to 51700 M - 1 CM -1 . Under dark conditions, in the presence of 100 μM of the target compound, the survival rates of Hela and 4T1 cells both reached over 90%, demonstrating its low dark toxicity. Under photoactivation conditions, the anti-proliferative inhibitory activity IC 50 of some target compounds against Hela and 4T1 cells was 0.61 - 15.85 μM, demonstrating its good photosensitizing anti-tumor effect. In addition, the results of the co-localization experiment in cells showed that in 4T1 cells, QMA10 could spontaneously target the endoplasmic reticulum, and QM10 exhibited good in vivo anti-tumor activity.

[0267] Therefore, the compounds involved in the present invention have the properties of aggregation-induced fluorescence enhancement, extremely low dark toxicity, and excellent phototoxicity, and are expected to be developed into a novel class of photosensitizers for photodynamic therapy, with good anti-tumor application prospects.

[0268] After considering the specification and practicing the content disclosed herein, those skilled in the art will readily conceive of other embodiments of the present application. The present application is intended to cover any variations, uses, or adaptations of the present application, which follow the general principles of the present application and include the common general knowledge or conventional technical means in the technical field not disclosed in the present application.

[0269] It should be understood that the present application is not limited to the exact structures described above and shown in the drawings, and various modifications and changes can be made without departing from its scope.

Claims

1. A photosensitizer based on a quinoline-malononitrile nucleus, characterized in that, It is a compound represented by the general formula (I): wherein R1 is H, OH, OTs, SCH3, I, Br, N3, NH2, R2 is H, CH3, OCH3 or F; The linker is 2. The photosensitizer based on the quinoline-malononitrile nucleus according to claim 1, characterized in that: R1 is I, Br, NH2, OH, R2 is H, CH3 or OCH3.

3. The photosensitizer based on quinoline-malononitrile nucleus according to claim 1, wherein, The compound represented by the general formula (I) or its pharmaceutically acceptable salt or its stereoisomer or its prodrug is selected from the following compounds: (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-ethylquinolin-4(1H)-ylidene)malononitrile, namely QM01; (E)-2-(2-(4-(Diphenylamino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM02; (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM03; (E)-2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM04; (E)-2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM05; (E)-2-(2-(2-(5-(4-(Diphenylamino)phenyl)thiophen-2-yl)vinyl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM06; (E)-2-(2-(2-(5-(4-(Di-p-toluidino)phenyl)thiophen-2-yl)vinyl)-1-(2-(methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM07; (E)-2-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-methylthio)ethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM08; (E)-2-(2-(4-(Diphenylamino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM09; (E)-2-(2-(4-(Di-p-toluidino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM10; (E)-2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM11; (E)-2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM12; (E)-2-(2-(2-(5-(4-(Diphenylamino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM13; (E)-2-(2-(2-(5-(4-(Di(p-tolyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM14; (E)-2-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM15; (E)-2-(2-(2-(5-(4-(Bis(4-fluorophenyl)amino)phenyl)thiophen-2-yl)vinyl)-1-(2-hydroxyethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM16; (E)-(2-(4-(Diphenylamino)-2-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethyl sulfonate, namely QM17; (E)-(2-(2-(4-(Di(p-tolyl)amino)styryl)4-(dicyanomethylene)quinolin-1(4H)-yl)ethyl)dimethyl sulfate, namely QM18; (E)-(2-(2-(4-(Bis(4-methoxyphenyl)amino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethyl sulfonate, namely QM19; (E)-(2-(2-(4-(Bis(4-fluorophenyl)amino)styryl)-4-(dicyanovinyl)quinolin-1(4H)-yl)ethyl)dimethyl sulfonate, namely QM20; (E)-(2-(4-(Dicyanomethylene)-2-(2-(5-(4-(diphenylamino)phenyl)thiophen-2-yl)vinyl)quinolin-1(4H)-yl)ethyl)dimethyl sulfonate, namely QM21; (E)-(2-(2-(5-(4-(Di(p-tolyl)amino)phenyl)thiophen-2-yl)vinyl)-4-(dicyanomethylene)quinolin-1(4H)-yl)ethyl)dimethyl sulfonate, namely QM22; (E)-(2-(2-(5-(4-(Bis(4-methoxyphenyl)amino)phenyl)thiophen-2-yl)vinyl)-4-(dicyanomethylene)quinolin-1(4H)-yl)ethyl)dimethyl sulfonate, namely QM23; (E)-2-(2-(4-(Di(p-tolyl)amino)styryl)-4-(dicyanomethylene)quinolin-1(4H)-yl)ethyl 4-methylbenzenesulfonate, namely QM24; (E)-2-(2-(4-(Di(p-toluidino)styryl)-1-(2-iodoethyl)quinolin-4(1H)-ylidene)malononitrile, namely QM25; (E)-2-(1-(2-bromoethyl)-2-(4-(Di(p-toluidino)styryl)quinolin-4(1H)-ylidene)malononitrile, namely QM26; (E)-2-(1-(2-azidoethyl)-2-(4-(Di(p-toluidino)styryl)quinolin-4(1H)-ylidene)malononitrile, namely QM27; (E)-2-(1-(2-aminoethyl)-2-(4-(di-p-tolylamino)styryl)quinolin-4(1H)-ylidene)malononitrile, namely QM28; (E)-(4-((2-(4-(di-p-toluidino)styryl)-4-(dicyanomethyl)quinolin-1(4H)-yl)ethyl)amino)-4-oxobutyl)triphenylphosphonium, namely QM29; (E)-1-(4-((2-(4-(di-p-tolylamino)styryl)-4-(dicyanomethyl)quinolin-1(4H)-yl)ethyl)amino)-4-oxobutyl)pyridin-1-ium, namely QM30.

4. The preparation method of the photosensitizer based on quinoline-malononitrile nucleus according to any one of claims 1-3, characterized in that, Comprising: i) Using 2-methylquinoline and bromide as starting materials, with toluene as the solvent, reacting at 110 °C to obtain the corresponding quinolinium salt; ii) Under the condition that the quinolinium salt uses sodium ethoxide as the base and ethanol as the solvent, reacting with malononitrile to obtain the corresponding quinoline-malononitrile nuclei Q1 and Q2; Q2 is obtained by OTs substitution and reaction with sodium methanethiolate to obtain Q3; iii) Q1, Q2, and Q3 undergo Knoevenagel condensation reaction with triphenylamine containing a mono-substituted aldehyde group to obtain photosensitizers based on the quinoline-malononitrile nucleus.

5. The preparation method according to claim 4, wherein: The photosensitizers QM02-QM08 containing a methylthio substitution react with methyl iodide and silver tetrafluoroborate to obtain the corresponding sulfonium-substituted photosensitizers QM17-QM23; The photosensitizer QM10 containing a hydroxyl substitution undergoes p-toluenesulfonylation of the hydroxyl group to obtain QM24, and subsequently further obtains bromine-, iodine-, and azide-substituted photosensitizer molecules QM25, QM26, and QM27 through substitution reactions; The azide-substituted photosensitizer molecule QM27 is reduced to obtain an amino-substituted photosensitizer molecule QM28, and further introduces a pyridinium salt and triphenylphosphonium through a condensation reaction to obtain the corresponding QM29 and QM30.

6. Use of the photosensitizer based on quinoline-malononitrile nucleus according to any one of claims 1-3 in the preparation of a medicament for treating or preventing tumors, characterized in that, The said compound or its pharmaceutically acceptable salt or its stereoisomer or its prodrug kills tumor cells by photodynamic therapy.

7. The application according to claim 6, wherein The said tumor cells are murine triple-negative breast cancer cells 4T1 or human cervical cancer cells Hela.

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