Method for directed photocatalytic synthesis and application of conjugated polymers

Abstract

The present invention relates to the field of organic synthesis and biomedicine, and discloses a method for directional photocatalytic synthesis and the use of conjugated polymers in the preparation of drugs for treating solid tumors. + Specifically converts it into 1,6-NADH, and uses the properties of the generated 1,6-NADH to regulate the original NAD in hypoxic tumor cells + / NADH balance, disrupting the mitochondrial electron respiratory chain and inhibiting the growth of solid tumors under hypoxic conditions.

Description

Technical Field

[0001] The present invention relates to the technical field of organic synthesis and biomedicine, and in particular to a method for directional photocatalytic synthesis and the application of conjugated polymers. Background Art

[0002] Nicotinamide adenine dinucleotide redox couple (NAD + / NADH) plays a very important coenzyme role in cell life activities, especially in the mitochondrial electron transport chain and glycolysis. Under natural physiological conditions, NAD + It will be oxidized into 1,4-NADH with coenzyme activity, and then further reduced to NAD during glycolysis and other processes. + In this process, ATP required for cell life activities is produced. Under external conditions, NAD + The oxidation of NAD often produces two products: 1,4-NADH and 1,6-NADH, which are difficult to separate due to their similar physical properties. + It is difficult to achieve single product oxidation of NAD, specifically producing 1,6-NADH to study + / NADH redox plays a very important role in the circulation of cells.

[0003] Conjugated polymers have attracted considerable research interest in chemistry, biology, and medicine due to their unique photophysical and photochemical properties. Their rigid, delocalized π-electron backbones impart strong light absorption / emission and signal transmission capabilities. Their highly reactive side chain groups allow conjugated polymers to possess a wide range of biological functions, enabling their widespread application in biomedicine.

[0004] Hypoxia in tumors is primarily due to the rapid proliferation of tumor cells. During this process, oxygen and energy are rapidly consumed, increasing internal tumor pressure and preventing oxygen from diffusing into the tumor from the outside, leading to a decrease in local oxygen levels. Compared to cells in normal normoxia, the hypoxic tumor microenvironment often exhibits elevated reactive oxygen species (ROS) levels, a low pH, and abnormal metabolic activity. Hypoxia can lead to tumor cell heterogeneity and suppress innate adaptive immune responses, thereby increasing the likelihood of tumor metastasis. Furthermore, some tumor cells can maintain a chronic hypoxic state, and some hypoxic tumor cells can even tolerate radiotherapy and chemotherapy. Photodynamic therapy (PDT) is used in cancer treatment due to its high efficacy and rapid response. However, due to a lack of oxygen, PDT cannot consistently generate sufficient ROS levels to kill tumors, limiting the development of PDT strategies. Therefore, a strategy that can sustain tumor cell killing under low or even no oxygen conditions is crucial. Summary of the Invention

[0005] The main purpose of the present invention is to provide a new method for achieving directed synthesis of compounds using water-soluble conjugated polymer materials and the application of the conjugated polymer in the preparation of drugs for treating solid tumors, by means of photocatalysis to convert NAD into + Specifically converts it into 1,6-NADH, and uses the characteristics of the generated 1,6-NADH to regulate the original NAD in hypoxic tumor cells + / NADH balance, disrupting the mitochondrial electron respiratory chain and inhibiting the growth of solid tumors under hypoxic conditions.

[0006] In order to achieve the above-mentioned object, the present invention provides, on one hand, the use of a conjugated polymer having a structure represented by formula (I) in the preparation of a drug for treating solid tumors.

[0007]

[0008] In formula (I), n is an integer from 10 to 20; Wherein, X is a halogen.

[0009] Preferably, the solid tumor is a breast tumor.

[0010] According to a second aspect of the present invention, a method for directional photocatalytic synthesis is provided, wherein the method comprises: + The step of irradiating the reaction mixture with visible light, wherein the structure of the conjugated polymer is as shown in formula (I),

[0011]

[0012] In formula (I), n is an integer from 10 to 20; Wherein, X is a halogen.

[0013] Preferably, the reaction mixture is prepared by the following steps:

[0014] 1) mixing the conjugated polymer with water to prepare a water-soluble conjugated polymer dispersion;

[0015] 2) reacting the water-soluble conjugated polymer dispersion with the electron sacrificial agent and the reaction raw material NAD + Contact is performed to obtain the reaction mixture.

[0016] Preferably, in step 1), the content of the conjugated polymer in the water-soluble conjugated polymer dispersion is 1-10 mmol / L.

[0017] Preferably, in step 2), the content of the conjugated polymer in the reaction mixture is 1-20 μmol / L.

[0018] Preferably, in step 2), the content of the electron sacrificial agent in the reaction mixture is 4-100 mmol / L.

[0019] Preferably, in step 2), the electron sacrificial agent is triethanolamine and / or ascorbic acid.

[0020] Preferably, in step 2), the reaction raw material NAD in the reaction mixture + The content is 0.01-5mmol / L.

[0021] Preferably, the illumination conditions include: illumination time of 20-180 minutes, illumination wavelength of 400-700 nm, illumination power density of 30-100 mW / cm 2 .

[0022] Through the above technical solution, the present invention has the following beneficial effects:

[0023] (1) The water-soluble conjugated polymer dispersion of the present invention can be photoregulated at room temperature and pressure to specifically catalyze the oxidation of NAD + Converted to 1,6-NADH, which specifically oxidizes NAD under physiological conditions + It plays an important role in studying the cellular oxidative respiratory chain and the cycle of coenzyme redox pairs during glycolysis.

[0024] (2) The present invention uses a simple photocatalytic method to prepare 1,6-NADH. Compared with photodynamic therapy, due to the lower oxygen level in the hypoxic state, it is impossible to continuously provide sufficient oxygen for photodynamic therapy to generate reactive oxygen species, thereby limiting the development of photodynamic strategies for treating cancer. The method of using 1,6-NADH to produce 1,6-NADH destroys the original NAD in the cell. + The balance of 1,6-NADH can effectively and continuously kill hypoxic tumor cells. The catalyst used in the present invention is independent of oxygen and can continuously produce 1,6-NADH in situ through light irradiation to kill tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 These are the absorption spectrum and fluorescence spectrum of the conjugated polymer in Example 1.

[0026] Figure 2 This is the photocurrent curve of the photoelectric response signal of the conjugated polymer in Example 1.

[0027] Figure 3 This is the ultraviolet photoelectron spectrum of the conjugated polymer in Example 1.

[0028] Figure 4 The calculated HOMO / LUMO orbitals and NAD of the conjugated polymers of the present invention + / NADH redox couple potential.

[0029] Figure 5 Absorption spectrum before and after illumination in Example 2

[0030] Figure 6 (A) represents the intracellular NAD before and after illumination in Example 2 + / NADH ratio changes.

[0031] Figure 6 (B) is a graph showing the survival rate of cells after adding the conjugated polymer and irradiating the cells in Example 2.

[0032] Figure 6 (C) is a graph showing changes in intracellular ATP levels before and after illumination in Example 2. DETAILED DESCRIPTION

[0033] The endpoints of the ranges and any values disclosed herein are not limited to the precise ranges or values, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0034] According to a first aspect of the present invention, there is provided use of a conjugated polymer having a structure represented by formula (I) in the preparation of a drug for treating solid tumors.

[0035]

[0036] In formula (I), n is an integer from 10 to 20; Wherein, X is a halogen.

[0037] According to the present invention, n may be, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or the like.

[0038] Preferably, X is fluorine, chlorine, bromine or iodine, more preferably fluorine, chlorine or bromine, particularly preferably bromine.

[0039] In the present invention, the conjugated polymer can be prepared according to the method disclosed in Chong H, Nie CY, Zhu CL, Yang Q, Liu LB, LvFT, Wang S, Langmuir, 2012, 28: 2091-2098.

[0040] According to the present invention, preferably, the solid tumor is a breast tumor.

[0041] The water-soluble conjugated polymer of the present invention can convert NAD into ATP in solution and in biological system under the irradiation of visible light xenon lamp. + It is specifically converted into 1,6-NADH, and the properties of the generated 1,6-NADH are used to regulate the original NAD in hypoxic tumor cells. + / NADH balance, thereby destroying the mitochondrial electron respiratory chain, reducing the mitochondrial membrane potential, disrupting the intracellular ATP balance, inducing cancer cell apoptosis, and achieving the purpose of treating solid tumors under hypoxic conditions.

[0042] According to a second aspect of the present invention, a method for directional photocatalytic synthesis is provided, wherein the method comprises: + The step of irradiating the reaction mixture with visible light, wherein the structure of the conjugated polymer is as shown in formula (I),

[0043]

[0044] In formula (I), n is an integer from 10 to 20; Wherein, X is a halogen.

[0045] According to the present invention, n may be, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or the like.

[0046] Preferably, X is fluorine, chlorine, bromine or iodine, more preferably fluorine, chlorine or bromine, particularly preferably bromine.

[0047] According to the present invention, preferably, the reaction mixture is prepared by the following steps:

[0048] 1) mixing the conjugated polymer with water to prepare a water-soluble conjugated polymer dispersion;

[0049] 2) reacting the water-soluble conjugated polymer dispersion with the electron sacrificial agent and the reaction raw material NAD + Contact is performed to obtain the reaction mixture.

[0050] The present invention effectively disperses the conjugated polymer in water, and the energy level of the conjugated polymer is closely related to NAD + The energy level of the conjugated polymer matches that of NADH, and it has strong light capture and photoelectric conversion capabilities. Under white light irradiation, the conjugated polymer can catalyze the conversion of NAD + Converted into 1,6-NADH.

[0051] According to the present invention, preferably, in step 1), the content of the conjugated polymer in the water-soluble conjugated polymer dispersion is 1-10 mmol / L, for example, it can be 1 mmol / L, 2 mmol / L, 3 mmol / L, 4 mmol / L, 5 mmol / L, 6 mmol / L, 7 mmol / L, 8 mmol / L, 9 mmol / L, 10 mmol / L, etc.

[0052] According to the present invention, preferably, in step 1), the mixing can be carried out at room temperature, for example, at 5-40° C., preferably at 10-30° C. The mixing time is not limited as long as the mixture is fully mixed, for example, it can be 20-60 minutes.

[0053] According to the present invention, preferably, in step 2), the content of the conjugated polymer in the reaction mixture is 1-20 μmol / L, for example, it can be 1 μmol / L, 2 μmol / L, 3 μmol / L, 4 μmol / L, 5 μmol / L, 6 μmol / L, 7 μmol / L, 8 μmol / L, 9 μmol / L, 10 μmol / L, 11 μmol / L, 12 μmol / L, 13 μmol / L, 14 μmol / L, 15 μmol / L, 16 μmol / L, 17 μmol / L, 18 μmol / L, 19 μmol / L, or 20 μmol / L.

[0054] According to the present invention, preferably, in step 2), the content of the electron sacrificial agent in the reaction mixture is 4-100 mmol / L, for example, it can be 4 mmol / L, 5 mmol / L, 6 mmol / L, 7 mmol / L, 8 mmol / L, 9 mmol / L, 10 mmol / L, 11 mmol / L, 12 mmol / L, 13 mmol / L, 14 mmol / L, 15 mmol / L, 16 mmol / L, 20 mmol / L, 30 mmol / L, 35 mmol / L, 40 mmol / L, 45 mmol / L, 50 mmol / L, 55 mmol / L, 60 mmol / L, 70 mmol / L, 80 mmol / L, 90 mmol / L, 100 mmol / L, etc.

[0055] In the present invention, preferably, in step 2), the electron sacrificial agent is triethanolamine and / or ascorbic acid, more preferably triethanolamine.

[0056] According to the present invention, preferably, in step 2), the reaction raw material NAD in the reaction mixture +The content is 0.01-5mmol / L, for example, it can be 0.01mmol / L, 0.02mmol / L, 0.03mmol / L, 0.04mmol / L, 0.05mmol / L, 0.06mmol / L, 0.07mmol / L, 0.08mmol / L, 0.09mmol / L, 0.1mmol / L, 0.2mmol / L, 0.3mmol / L, 0.4mmol / L, 0.5mmol / L, 0.6mmol / L, 0.7mmol / L, 0.8mmol / L, 0.9mmol / L, 1mmol / L, 1.5mmol / L, 2mmol / L, 2.5mmol / L, 3mmol / L, 3.5mmol / L, 4mmol / L, 45mmol / L, 5mmol / L, etc.

[0057] According to the present invention, preferably, the conditions for illumination include: illumination time of 20-180 minutes, illumination wavelength of 400-700 nm, illumination power density of 30-100 mW / cm 2 .

[0058] In a particularly preferred embodiment of the present invention, the illumination conditions include: illumination time of 20 minutes, illumination wavelength of 400-700 nm, illumination power density of 50 mW / cm 2 .

[0059] According to the present invention, the light source of visible light is a light source that can provide a wavelength of 400 nm to 700 nm, and can be, for example, a xenon lamp.

[0060] The present invention will be described in detail below through examples, but the present invention is not limited to the following examples.

[0061] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0062] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0063] The conjugated polymers used in the following examples were synthesized according to the method disclosed in Chong H, Nie CY, Zhu CL, Yang Q, Liu LB, Lv FT, Wang S, Langmuir, 2012, 28: 2091-2098. The specific structure of the conjugated polymer (hereinafter referred to as PFP) is as follows:

[0064]

[0065] Where n is 12; Wherein, X is Br.

[0066] Example 1

[0067] 1. Preparation of water-soluble conjugated polymer dispersion

[0068] (1) Disperse 7 mg of conjugated polymer powder in 1 mL of DMSO and ultrasonicate for 1 h to obtain a 10 mM polymer stock solution.

[0069] (2) Add 9 mL of deionized water and continue ultrasonication for 1 h.

[0070] 2. Characterization

[0071] (1) Absorption and fluorescence spectra of conjugated polymers

[0072] 1 mM conjugated polymer was diluted in aqueous solution to prepare a 10 μM solution. The absorption and fluorescence spectra of the conjugated polymer were measured at a constant temperature of 25°C (absorption spectra were measured using a Thermo Scientific Evolution 201; fluorescence spectra were measured using a Hitachi F-4500 fluorometer). Figure 1 .

[0073] pass Figure 1 It can be seen that the conjugated polymer has obvious absorption in the visible light range. The band gap E can be calculated according to the following formula: g It is 2.46eV.

[0074]

[0075] λ g : Absorption wavelength threshold

[0076] (2) Testing the photocurrent response of conjugated polymers

[0077] Solution preparation: 10 μM PFP aqueous solution, 100 mM TEOA aqueous solution (triethanolamine aqueous solution), and 0.5% by volume Nafion aqueous solution were prepared respectively.

[0078] 30 μL of 1mM PFP aqueous solution was added dropwise onto 1.2cm×1.2cm carbon paper and placed in a fume hood to allow the solvent to evaporate naturally. 20 μL of Nafion aqueous solution was added dropwise to fix the material on the carbon paper electrode. The carbon paper treated as above was used as the working electrode, Ag / AgCl as the reference electrode, and the Pt electrode as the counter electrode. They were placed in an electrolytic cell and the photoelectric response was tested using an electrochemical workstation (Metrohm, Autolab PGATAT 302N, Switzerland) (the electrolyte was an aqueous solution containing 1mM triethanolamine, and the light source was white light). The applied voltage was 0.2V, the scanning speed was 0.05mV / s, and the graph was obtained. Figure 2 .

[0079] pass Figure 2 It can be seen that the conjugated polymer has an obvious photocurrent response.

[0080] (3) Ultraviolet photoelectron spectroscopy analysis to test the highest occupied molecular orbital of conjugated polymers

[0081] A 1 mM aqueous solution of the conjugated polymer was dropped onto a 1 cm × 1 cm ITO film and a film was prepared using a spin coater at a speed of 2000 rpm for 2 min. The highest occupied molecular orbital was calculated using a photoelectron spectrometer (Kratos, model AXIS ULTRA DLD) and plotted. Figure 3 .

[0082] pass Figure 3 The test results show that the HOMO of the conjugated polymer is -5.82 eV and the LUMO is -3.36 eV using the following formula.

[0083] E HOMO =-[21.22-(E cutoff -E F )]

[0084] E HOMO =E g +E LUMO

[0085] E Cutoff : Low kinetic energy cut-off edge

[0086] E F : High kinetic energy starting edge

[0087] Based on the calculated band gap and LUMO energy, it is speculated that when the PFP electrons are excited by light, electron / hole separation will occur, the electrons will jump from the HOMO orbital to the LUMO orbital, and then further transfer to NAD+ with a potential of -0.54V, and the holes will be captured by triethanolamine. Based on this process and the energy calculated above, the graph is obtained Figure 4 .

[0088] Example 2 (Application Example)

[0089] 4T1 cells were purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, with the catalog number 3131C0001000800032.

[0090] RPMI 1640 culture medium was purchased from Thermo Fisher, product number 22400105.

[0091] 1. Conjugated polymers convert NAD + Catalytic oxidation to 1,6-NADH

[0092] Add PFP aqueous solution, TEOA aqueous solution and NAD into the quartz cell + (β-nicotinamide adenine dinucleotide, purchased from Sigma, catalog number 10127973001), and supplemented to 2 mL with deionized water so that PFP was 10 μM, TEOA was 100 mM, NAD + For 2mM, use 50mW / cm 2 The quartz cell was irradiated with white light for 30 minutes, and its absorption spectrum was measured after the reaction was completed. Figure 5 .

[0093] Figure 5 The absorption spectrum before and after illumination in Example 2 is shown in FIG. Figure 5 It can be seen that after illumination, the absorption of the reaction system at 345 nm is significantly enhanced, and the maximum absorption position is at 345 nm, which matches the characteristic absorption of 1,6-NADH, and the product is 1,6-NADH.

[0094] 2. Intracellular NAD + Content level test

[0095] Take 5×10 5 / mL 4T1 cells were seeded into six-well plates and cultured for 24 hours to allow the cells to adhere (4T1 cells were cultured in RPMI 1640 culture medium containing 10% FBS by volume, placed in a cell culture incubator at 5% CO2 and 37°C. The cells were digested with 0.25% trypsin for 2 minutes and subcultured at a ratio of 1:4.) The supernatant culture medium was discarded and replaced with a cell culture medium containing 8μM PFP and 1mM TEOA and incubated overnight. The supernatant was discarded, the cells were washed three times with PBS, replaced with a cell culture medium containing 1mM TEOA, and the cells were placed in a cell culture incubator at 5% CO2, 1% O2, and 37°C and incubated for another 6 hours. At 50mW / cm 2The supernatant was discarded and 200 μL of cell lysis buffer (purchased from Beyotime Biotechnology, Cat. No. P0013) was added to each well. The cells were collected and centrifuged at 4°C, 1200 rpm for 5 minutes. The supernatant was collected for detection.

[0096] 25 μL of sample solution was added to a 96-well plate and the NAD + / NADH ratio kit (purchased from Solebol, Cat. No. 15263) to test its NAD + The ratio is obtained Figure 6 (A) in the.

[0097] Figure 6 (A) represents the intracellular NAD before and after illumination in Example 2 + / NADH ratio changes. Figure 6 As shown in (A), under light conditions, the NAD + The proportion will decrease, indicating that the NAD in the cell + Photocatalyzed by PFP to 1,6-NADH.

[0098] 3. Analysis of phototoxicity experiments after conjugated polymer PFP acts on cells

[0099] 4T1 cells were cultured in RPMI 1640 medium containing 10% FBS in a cell culture incubator at 5% CO2 and 37°C. The cells were digested with 0.25% trypsin for 2 minutes and subcultured at a 1:4 ratio.

[0100] The cell viability was analyzed using the MTT assay. 100 μL of 8 × 10 4 Cells were seeded into 96-well flat-bottom cell culture plates and cultured overnight. The supernatant was discarded and replaced with RPMI 1640 medium containing 8 μM PFP and 1 mM TEOA. After culturing for 24 hours, the cells were placed in a cell culture incubator at 1% O2, 5% CO2, and a temperature of 37°C for another 6 hours. 2 Irradiate under white light for 20 minutes. Subsequently, discard the supernatant solution and replace it with RPMI 1640 culture medium containing 10% FBS by volume, and culture it in a cell culture incubator with 5% CO2 and a temperature of 37°C for 40 hours. Replace the original culture medium with the corresponding cell culture medium containing 0.mg / mL MTT, and continue to culture for 4 hours at 5% CO2 and 37°C. Discard the upper liquid, add 100μL DMSO to each well to dissolve the blue-purple reduction product formazan, and measure the absorption at 570nm with an enzyme marker. The cell survival rate is as follows: Figure 6 As shown in (B) in the figure. Figure 6As shown in (B), the conjugated polymer has no dark toxicity to cells when the concentration is not higher than 8 μM.

[0101] The cell viability was calculated according to the following formula:

[0102]

[0103] In formula (1),

[0104] A: absorbance value of the experimental group, A0: absorbance value without any treatment,

[0105] A b : Absorbance value of blank well.

[0106] 4. Intracellular ATP content level test

[0107] Take 8×10 4 / mL 4T1 cells were seeded into six-well plates and cultured for 24 hours to allow the cells to adhere (4T1 cells were cultured in RPMI 1640 culture medium containing 10% FBS by volume, placed in a cell culture incubator at 5% CO2 and 37°C. The cells were digested with 0.25% trypsin for 2 minutes and subcultured at a ratio of 1:4.) The supernatant culture medium was discarded and replaced with a cell culture medium containing 8μM PFP and 1mM TEOA and incubated overnight. The supernatant was discarded, the cells were washed three times with PBS, replaced with a cell culture medium containing 1mM TEOA, and the cells were placed in a cell culture incubator at 5% CO2, 1% O2, and 37°C and incubated for another 6 hours. At 50mW / cm 2 The supernatant was discarded and 200 μL of cell lysis buffer (purchased from Beyotime Biotechnology, Cat. No. P0013) was added to each well. The cells were collected and centrifuged at 4°C, 1200 rpm for 5 minutes. The supernatant was collected for detection.

[0108] Preparation of working solution: Mix ATP detection reagent and ATP detection reagent diluent (ATP kit purchased from Beyotime Biotechnology, product number S0027) in a ratio of 1:4 and let it stand at room temperature for 3-5 minutes.

[0109] Add 20 μL of sample and 100 μL of working solution to a 96-well plate, and measure the chemiluminescence at 540 nm with an enzyme reader every two minutes to obtain Figure 6 (C) in.

[0110] Figure 6 (C) is a graph showing the changes in intracellular ATP levels before and after illumination in Example 2. Figure 6 As shown in (C), after illumination, the ATP level of cells with conjugated polymers decreased significantly, indicating that the NAD +The balance of NADH / NADH is broken, which leads to damage of mitochondrial respiratory chain and further decrease of ATP level.

[0111] It can be seen from the above examples 1-2 that the conjugated polymer of the present invention has a strong light-harvesting ability, and its energy level is similar to that of NAD. + The energy level of the catalytic oxidation reaction to 1,6-NADH matches. After the conjugated polymer is excited by light, it transfers electrons to NAD + , synthesize 1,6-NADH molecules, and using the characteristics of 1,6-NADH, the catalyst can photocatalytically oxidize NAD in an oxygen-deficient environment + After being converted to 1,6-NADH, it destroys the original NAD in the cell + / NADH balance, destroying the mitochondrial electron transport chain, affecting the normal metabolic function of cells, and inducing apoptosis of hypoxic cancer cells.

[0112] The present invention plays an important role in developing the specific photocatalytic synthesis of the catalytic system of conjugated polymers and provides a new method for treating hypoxic tumors.

[0113] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as disclosed in the present invention and fall within the scope of protection of the present invention.

Claims

1. A method for directional photocatalytic synthesis, characterized in that: The method comprises: making a conjugated polymer, an electron sacrificial agent and a reaction raw material NAD + The reaction mixture is irradiated with visible light, wherein the structure of the conjugated polymer is as shown in formula (I), Formula (I) In formula (I), n is an integer of 10 to 20; R= , wherein X is a halogen.

2. The method according to claim 1, wherein The reaction mixture is prepared by the following steps: 1) mixing the conjugated polymer with water to prepare a water-soluble conjugated polymer dispersion; 2) reacting the water-soluble conjugated polymer dispersion with the electron sacrificial agent and the reaction raw material NAD + Contact is performed to obtain the reaction mixture.

3. The method according to claim 2, wherein: In step 1), the content of the conjugated polymer in the water-soluble conjugated polymer dispersion is 1-10 mmol / L.

4. The method according to claim 2, wherein: In step 2), the content of the conjugated polymer in the reaction mixture is 1-20 μmol / L.

5. The method according to any one of claims 1 to 4, wherein: In step 2), the content of the electron sacrificial agent in the reaction mixture is 4-100 mmol / L.

6. The method according to claim 5, wherein: In step 2), the electron sacrificial agent is triethanolamine and / or ascorbic acid.

7. The method according to any one of claims 1 to 4, wherein: In step 2), the reaction raw material NAD in the reaction mixture + The content is 0.01-5mmol / L.

8. The method according to any one of claims 1 to 4, wherein: The illumination conditions include: illumination time of 20-180 minutes, illumination wavelength of 400-700nm, illumination power density of 30-100 mW / cm 2 .

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