Method for photoinduced preparation of aromatic propiopyrrone and biological applications

Abstract

The application discloses a method for preparing aromatic benzyl propargyl ketone by light induction and biological application, belongs to the technical field of organic synthesis and biological medicine, and converts aromatic benzyl propargyl alcohol into aromatic benzyl propargyl ketone under the condition of light induction; the change of the functional group before and after light induction is that the hydroxyl alkyne without reaction activity is converted into the carbonyl alkyne with click reaction activity, fluorescence enhancement can be realized, and the method can be used for imaging and tracing; and the aromatic benzyl propargyl ketone can react with a compound containing an amino group or a biological macromolecule to realize the killing of tumor cells. The method uses light as a starting switch, the reaction condition is simple, controllability is good, no additional catalyst is needed, and the method is easy to mass-produce, so the method has wide application prospects in polypeptide or protein labeling, anti-tumor and the like.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of organic synthesis and biological medicine, and particularly relates to a method for preparing aromatic benzenopropynone by photoinduction and biological application. BACKGROUND

[0002] Cancer is one of the most common diseases worldwide, and its morbidity and mortality are high, which is a major public health problem worldwide. It not only seriously affects the life quality and life span of patients, but also has a profound impact on medical resources and social economy. Chemotherapy is one of the most widely used cancer treatment methods in clinical practice, and can effectively inhibit tumor proliferation and metastasis in the early stage of treatment. However, with the increase of treatment period, many patients will have drug resistance problems. In order to solve this problem, the current clinical method is to passively increase the dosage. However, increasing the dosage often increases the toxic side effects of chemotherapeutic drugs on normal cells. Therefore, it is crucial to achieve the site-specific activation of chemotherapeutic drugs at the lesion site for the further development of chemotherapy.

[0003] The patent document with the publication number CN101500557A discloses the synthesis and use of nitrofurans (such as nitrofurantoin) for treating cancer and inhibiting angiogenesis, and the structural formula of the nitrofurans is represented as: The compound can be delivered by liposomes, and the liposomes can be targeted to specific tissues by coupling with specific ligands such as sugars, glycolipids or proteins; the patent document with the publication number CN108451939A discloses the use of 2,4-dinitrobenzenesulfonamide compounds for preparing drugs for treating cervical cancer, and the 2,4-dinitrobenzenesulfonamide compounds achieve targeted cancer treatment by inhibiting the activity of thioredoxin reductase. However, the above-mentioned compounds cannot be activated at the lesion site.

[0004] Light, as an external stimulation signal, has the characteristics of simple operation, high resolution, low invasiveness and spatiotemporal controllability, and is widely used in the field of biomedicine. By using light with high spatiotemporal controllability and combining the in-situ nature of chemical reactions, a photochemical reaction system is constructed for site-specific activation of chemotherapeutic drugs at the tumor site, which can provide a new idea for the chemotherapy of tumors. SUMMARY

[0005] In order to overcome the deficiencies in the prior art, the application provides a method for preparing aromatic benzenopropynone by photoinduction and biological application.

[0006] The specific technical solutions adopted are as follows:

[0007] A method for preparing aromatic benzenopropargyl ketone by photoinduction, comprising: converting aromatic benzenopropargyl alcohol shown in formula (I) into aromatic benzenopropargyl ketone shown in formula (II) under photoinduction condition.

[0008]

[0009] In formula (I) and formula (II), R1 is a light absorption adjusting group, and is independently selected from phenyl, naphthyl, substituted phenyl, arylamine group or heteroaromatic ring group containing at least one nitrogen, oxygen or sulfur atom.

[0010] The inventors find that aromatic benzenopropargyl alcohol shown in formula (I) can be converted into aromatic benzenopropargyl ketone shown in formula (II) under photoinduction condition; the change of functional groups (conversion of hydroxyl alkyne without reaction activity into carbonyl alkyne with click reaction activity) before and after photoinduction can realize fluorescence enhancement, and can be used for imaging and tracking; and the aromatic benzenopropargyl ketone can react with compounds containing amino groups or biological macromolecules to realize killing of tumor cells.

[0011] Further, in the substituted phenyl, the substituent group is methoxy, alkylamine group, carbazole group, phenoxazine group, naphthylamine group or triphenyl ethenyl group.

[0012] Further, the arylamine group is diphenylamine group, triphenylamine group or dipyridylamine group.

[0013] Further, the heteroaromatic ring group containing at least one nitrogen, oxygen or sulfur atom is thienyl group, pyridyl group, furanyl group, carbazole group, phenothiazine group, phenoxazine group, phenothiazine phenyl group, bithiophenyl group, thienocyclopentadienyl group, 9,10-dihydro-9,9-dimethyl acridyl group, 9,10-dihydro-9,9-diphenyl acridyl group, 10-H-spiro[acridine-9,9'-fluorene] group, diphenylamine thienyl group, bithiophenyl group, fused thiophene group or thienocyclopentadienyl group.

[0014] It should be noted that R1 is a light absorption adjusting group, and its main function is to adjust the ultraviolet-visible-near infrared absorption of the molecule. By replacing different R1, the molecule can have different absorption, so that different light can be used to excite aromatic benzenopropargyl alcohol shown in formula (I) to convert into aromatic benzenopropargyl ketone shown in formula (II); but the real protein / polypeptide labeling effect and tumor cell killing effect is realized by the conversion of hydroxyl alkyne without reaction activity into carbonyl alkyne with click reaction activity after photoinduction.

[0015] Preferably, the photoinduction condition is irradiation under ultraviolet light or white light for ≥2s, further irradiation for ≥1min, and still further irradiation for ≥10min; the wavelength range of the ultraviolet light is 365nm-460nm; and the wavelength range of the white light is 400nm-1000nm.

[0016] More preferably, the power density of the ultraviolet light or white light irradiation is 5-200 mW cm -2 .

[0017] Optionally, the aromatic phenylpropynol shown in formula (I) is prepared by the following method:

[0018] (1) the compound and 5-bromo-2-nitrobenzaldehyde are heated to 60-80℃ under inert gas atmosphere to perform Suzuki reaction for 6-24 hours, and after the reaction is completed, the nitrobenzaldehyde derivative is obtained through post-treatment; the definition of R1 is the same as above;

[0019] (2) the organic solution of the nitrobenzaldehyde derivative is added with Grignard reagent under ice bath condition, and reacted for 2-6 hours under inert gas atmosphere, and after the reaction is completed, the aromatic phenylpropynol shown in formula (I) is obtained through post-treatment.

[0020] Specifically, in step (1), the molar ratio of the compound and 5-bromo-2-nitrobenzaldehyde is 1:1.5-2; the inert gas atmosphere is nitrogen atmosphere, and a catalyst and an auxiliary agent are further added during the Suzuki reaction;

[0021] Further, the catalyst is preferably Pd(PPh3)4, and the auxiliary agent is preferably CsCO3.

[0022] Specifically, in step (2), the Grignard reagent is ethynyl magnesium bromide, and the molar ratio of the nitrobenzaldehyde derivative and the ethynyl magnesium bromide is 1:1.2-2; the inert gas atmosphere is nitrogen atmosphere.

[0023] Preferably, the organic solvent for dissolving the nitrobenzaldehyde derivative is selected from tetrahydrofuran.

[0024] The present application also provides a compound, which is aromatic phenylpropynol or aromatic phenylpropynone, the structural formula of the aromatic phenylpropynol is shown in formula (I), and the structural formula of the aromatic phenylpropynone is shown in formula (II), and the definition of R1 is the same as above.

[0025] The present application also provides the use of the method for preparing the aromatic phenylpropynone under light induction or the compound in the preparation of a tumor treatment drug.

[0026] Preferably, the tumor treatment drug contains nanoparticles, the nanoparticle component contains the aromatic phenylpropynol shown in formula (I), and the nanoparticle is used to convert the aromatic phenylpropynol shown in formula (I) into the aromatic phenylpropynone shown in formula (II) under the condition of light induction to play a tumor treatment role.

[0027] Further preferably, the aromatic propargyl alcohol of structural formula (I) is mixed with a surfactant to prepare the nanoparticles by co-precipitation.

[0028] The application also provides the use of the method for preparing the aromatic propargyl ketone under light induction or the compound in the preparation of a polypeptide or protein fluorescent labeling reagent.

[0029] Preferably, the aromatic propargyl alcohol of structural formula (I) is contacted with a polypeptide or protein, and under light induction, the aromatic propargyl alcohol of structural formula (I) is converted into the aromatic propargyl ketone of formula (II) to play a fluorescent labeling role.

[0030] Compared with the prior art, the application has the following beneficial effects:

[0031] (1) The application converts the aromatic propargyl alcohol into the aromatic propargyl ketone by using the method of light induction, uses light as a starting switch, has good spatiotemporal response properties, and compared with the current synthesis method of the propargyl ketone derivative, the reaction condition of the application is simple, no additional catalyst is needed, and the application is easy to mass-produce.

[0032] (2) The reaction of converting the aromatic propargyl alcohol into the aromatic propargyl ketone under light induction in the application can be used for in-situ treatment of tumors, delivers the inactivated aromatic propargyl alcohol to a tumor lesion site, and makes it activated into the active aromatic propargyl ketone under light irradiation for the treatment of tumors. Compared with the characteristics that the photodynamic therapy needs oxygen, the application adopts the strategy of generating active substances in-situ, and exhibits a better treatment effect under anoxic conditions, and has a broad application prospect in cancer treatment.

[0033] (3) The aromatic propargyl alcohol / aromatic propargyl ketone in the application can be used for fluorescent labeling of proteins or polypeptides, overcomes the disadvantage that biological macromolecules have no fluorescence, and is used for cell imaging and tracking. In addition, the change of the functional group before and after light irradiation can also realize fluorescence enhancement of the compound. BRIEF DESCRIPTION OF DRAWINGS

[0034] Figure 1 It is an ultraviolet-visible absorption spectrum of the NTYOL molecule under different light irradiation times in dichloromethane solution.

[0035] Figure 2 It is a fluorescence emission spectrum of the NTYOL molecule under different light irradiation times in dichloromethane solution.

[0036] Figure 3 It is an SDS-PAGE result diagram of BSA-NSTYONE obtained after labeling BSA with NTYOL.

[0037] Figure 4Figure of SDS-PAGE results of NTYOL after intracellular protein labeling light irradiation.

[0038] Figure 5 Figure of 4T1 cell dark toxicity and phototoxicity evaluation results of NTYOL and crystal violet (CV) under normoxic conditions.

[0039] Figure 6 Figure of 4T1 cell dark toxicity and phototoxicity evaluation results of NTYOL and crystal violet (CV) under hypoxic conditions.

[0040] Figure 7 Figure of A, killing evaluation results of NTYOL NPs on 4T1 solid tumors in mice, and B, biological safety evaluation results of NTYOL NPs on model mice. DETAILED DESCRIPTION

[0041] The present application will be further clarified by the following examples and figures. It should be understood that these examples are intended to illustrate the present application and are not intended to limit the scope of the present application. The methods of operation used in the following examples, if not specified otherwise, are usually carried out in accordance with conventional methods or as recommended by the manufacturer. The contents not described in detail in the present specification belong to the prior art known to those skilled in the art. The experimental materials used in the following examples, if not specifically stated, can be purchased from conventional biochemical reagent companies.

[0042] Example 1

[0043]

[0044] Synthesis of 4-nitro-4'-(1,2,2-triphenylvinyl)-[1,1'-biphenyl]-3-carbaldehyde (NTC): 4,4,5,5-tetramethyl-2-(4-(1,2,2-triphenylvinyl)phenyl)-1,3,2-dioxaborinane (916 mg, 2 mmol), 5-bromo-2-nitrobenzaldehyde (690 mg, 3 mmol), Pd(PPh3)4 (60 mg, 0.1 mmol) and CsCO3 (489 mg, 3 mmol) were sequentially added into a 100 ml two-necked flask, nitrogen was introduced, pumped out, and nitrogen was introduced for 3 times (10 minutes each time), and then a pre-deaerated mixture of tetrahydrofuran / water was injected into the reaction flask. Then heated to 80°C, and reacted for 12 hours. After the reaction was completed, extracted with dichloromethane / water (the volume ratio of dichloromethane to water was 1 / 1), dried over anhydrous magnesium sulfate for 2 hours, and purified by column chromatography to obtain yellow solid NTC (722 mg) with a yield of 75%.

[0045] The characterization results of compound NTC are as follows:

[0046] 1 H NMR (500 MHz, Chloroform-d) δ 10.50 (s, 1H), 8.18 (d, J = 8.5 Hz, 1H), 8.08 (d, J = 2.1 Hz, 1H), 7.87 (dd, J = 8.5, 2.1 Hz, 1H), 7.41 (d, J = 8.5 Hz, 2H), 7.18 - 7.02 (m, 17H).

[0047] 13 C NMR (101 MHz, CDC13) δ 188.5, 147.8, 146.9, 145.4, 143.4, 143.3, 139.8, 135.2, 132.3, 131.3, 131.3, 131.3, 131.2, 127.9, 127.9, 127.7, 127.6, 126.8, 126.7, 126.7, 126.6, 125.3.

[0048] Example 2

[0049]

[0050] Synthesis of compound 1-(4-nitro-4'-(1,2,2-triphenylvinyl)-[1,1'-biphenyl]-3-yl)prop-2-yn-1-ol (NTYOL): In a 50 mL dry round bottom flask, 4-nitro-4'-(1,2,2-triphenylvinyl)-[1,1'-biphenyl]-3- carboxaldehyde NTQ (960 mg, 2 mmol) prepared in Example 1 was added, nitrogen was purged- aspirated- nitrogen was purged for 3 times (10 minutes each), then tetrahydrofuran (10 mL) previously degassed was injected into the reaction flask. Then, under ice bath conditions, a solution of ethynyl magnesium bromide in tetrahydrofuran (6 mL, molar ratio of ethynyl magnesium bromide to NTYOL about 2:1) was slowly injected into the reaction flask with a syringe, the reaction was left for 3 h. After stopping the reaction, saturated ammonium chloride solution was added to quench the reaction, extraction was carried out with dichloromethane / water (volume ratio of dichloromethane to water 1 / 1), dried over anhydrous magnesium sulfate for 2 hours, purified by column chromatography, obtaining NTYOL 0.91 g as a golden yellow solid, with a yield of 90%.

[0051] The characterization results of compound NTYOL are as follows:

[0052] 1H NMR (400 MHz, Chloroform-d) δ 8.12 (d, J = 1.7 Hz, 1H), 8.07 (d, J = 8.5 Hz, 1H), 7.65 (dd, J = 8.5, 1.9 Hz, 1H), 7.40 (d, J = 8.3 Hz, 2H), 7.19 - 7.01 (m, 17H), 6.08 (s, 1H), 2.64 (s, 1H).

[0053] 13 C NMR (101 MHz, CDC13) δ 146.7, 146.4, 145.0, 143.6, 143.5, 142.1, 140.1, 136.1, 135.7, 132.3, 131.5, 131.5, 131.4, 128.01, 127.96, 127.8, 127.8, 127.5, 126.9, 126.8, 126.8, 126.7, 81.6, 75.3, 61.6.

[0054] Example 3

[0055]

[0056] Synthesis of compound 1-(4-nitro-4'-(1,2,2-triphenylvinyl)-[1,1'-biphenyl]-3-yl)prop-2-yn-1-one (NSTYONE): In a 100 mL dry round bottom flask was added 1-(4-nitro-4'-(1,2,2-triphenylvinyl)-[1,1'-biphenyl]-3-yl)propan-2-yn-1-ol NTYOL (507 mg, 1 mmol) prepared in example 2, 30 mL of dichloromethane and irradiated with a 410 nm UV lamp (10 mW cm -2 ) for 1 h. After stopping the reaction the solvent was removed with a rotary evaporator and purified by column chromatography to obtain NSTYONE as a yellow solid 200 mg with a yield of 40%.

[0057] The results of the characterization of compound NSTYONE are as follows:

[0058] 1 H NMR (400 MHz, Chloroform-d) δ 8.12 (d, J = 1.7 Hz, 1H), 8.07 (d, J = 8.5 Hz, 1H), 7.65 (dd, J = 8.5, 1.9 Hz, 1H), 7.40 (d, J = 8.3 Hz, 2H), 7.19 - 7.01 (m, 17H), 6.08 (s, 1H), 2.64 (s, 1H).

[0059] 13C NMR (101 MHz, CDC13) δ 175.9, 146.4, 145.9, 145.3, 143.3, 143.2, 142.1, 139.8, 135.1, 134.8, 132.3, 131.2, 130.1, 127.9, 127.7, 126.8, 126.7, 126.6, 125.0, 82.1, 80.1.

[0060] During the above reaction, the reaction was dynamically monitored by using ultraviolet absorption spectrum and fluorescence emission spectrum. From the ultraviolet absorption spectrum of NTYOL, it can be seen that the absorption peak of the raw material NTYOL at 350 nm is obviously weakened with the increase of illumination time; when the illumination time is 1 min, the absorption peaks at 310 nm and 390 nm are obviously higher, and with the extension of illumination time, the absorption gradually red shifts, which is consistent with the ultraviolet absorption position of the product NSTYONE, so the change of ultraviolet absorption spectrum can be used to dynamically monitor the occurrence of the reaction. Figure 1

[0061] At the same time, the fluorescence emission spectrum of NTYOL at different illumination times was collected, and the fluorescence intensity (I) was tested and compared with the fluorescence intensity (I0) of NTYOL before illumination. As shown in Table 1, with the increase of illumination time, the fluorescence of the reaction system also showed obvious enhancement. Figure 2

[0062] Example 4

[0063] Using the property that aromatic phenylpropynol can be converted into aromatic phenylpropynone which can react with amino under light conditions, the bovine serum albumin was modified, and the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiment was used to verify that the bovine serum protein was modified and endowed with the property of natural protein fluorescence.

[0064] Modification of BSA (BSA-NSTYONE): In a 50 mL single-neck flask, 30 mg of bovine serum albumin BSA was added, then 2 mL of water was used to completely dissolve it, then 1 mL of 1-(4-nitro-4'-(1,2,2-triphenylvinyl)-[1,1'-biphenyl]-3-yl)prop-2-yn-1-ol NTYOL (5 mg) dissolved in 1 mL of tetrahydrofuran was added, and the reaction was carried out under the illumination of white light (400 nm-1000 nm, 40 mW cm -2 ) at room temperature for 30 min, then the cap was opened and stirred overnight, and the tetrahydrofuran was completely volatilized. The solvent water was removed by using a freeze dryer, and 20 mg of light yellow solid was obtained. As a control experiment, the same operation was carried out without illumination, and BSA+NTYOL was obtained.

[0065] ​​0.2 mg of standard bovine serum albumin BSA, 0.5 mg of BSA+NTYOL (NTYOL group) and 0.5 mg of BSA-NSTYONE (NTYOL+L group) were respectively weighed and dissolved in 1 mL (0.5 mol L -1 , pH 6.8) Tris-hydrochloric acid buffer or distilled water, and then subjected to SDS-PAGE experiment and fluorescence gel imaging characterization to prepare the sample to be tested. The sample to be tested was placed on the bottom plate of a fluorescence gel imager, and an excitation wavelength of 400 nm and an emission wavelength receiving filter of 530 nm were selected to form an imaging channel. After adjusting the focal length, a photograph was taken. The film after fluorescence gel imaging was placed in a large culture dish for staining, and 0.25% coomassie brilliant blue staining solution was used for staining for 1 h. The staining solution was discarded, and the film was rinsed with distilled water several times, and then a decolorizing solution was added for diffusion decolorization. The decolorizing solution was frequently replaced until the protein band was clear.

[0066] Subsequently, the protein band was placed on the bottom plate of a fluorescence gel imager, and a white light field was selected. After adjusting the focal length, a photograph was taken. As shown in Figure 3 the SDS-PAGE results, under ultraviolet light, BSA-NSTYONE (NTYOL+L group) can see a clear fluorescent band at the position of a molecular weight slightly lower than 70 KDa, while BSA+NTYOL (NTYOL group) has no fluorescence, proving that NTYOL and bovine serum protein can be successfully coupled under light conditions, and the natural protein can be endowed with the property of fluorescence by this coupling method.

[0067] Example 5

[0068] By utilizing the property that aromatic phenylpropynol can be converted into aromatic phenylpropynone which can react with amino under light conditions, the intracellular protein can be labeled, and the feasibility of this method can be verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) experiment.

[0069] Intracellular protein labeling: mouse breast cancer 4T1 cells were cultured in a cell culture dish. After the cells were completely adherent, the original complete culture medium was aspirated, and complete culture medium containing NTYOL (4 μM) was added, and the cells were cultured for another 2 h. Subsequently, the culture dish was placed under white light for 20 min (light power density was 40 mW cm -2 ). Subsequently, the culture was continued for 2 h. The cells in the culture dish were collected, and cell lysis solution was added for lysis treatment to extract the protein in the cells for treatment (NTYOL+L group). As a control experiment, the same operation was performed without light (NTYOL group).

[0070] SDS-PAGE experiments were performed following the steps in Example 4. The protein bands corresponding to the cellular proteins extracted in the above steps were placed on the substrate of a fluorescence gel imaging system, using a white light field. The focus was adjusted before taking the image. Figure 4 The SDS-PAGE results showed that there was no significant difference under white light. Under fluorescence, only the protein with added NTYOL (NTYOL group) did not show obvious fluorescence emission. However, after the protein with added NTYOL was irradiated with white light for 20 min (NTYOL+L group), it showed obvious fluorescence emission under fluorescence, indicating that this strategy can successfully label intracellular proteins.

[0071] Example 6

[0072] The photoinducible bioactivity of aromatic phenylpropynyl alcohol was utilized for hypoxic tumor killing. Using mouse breast cancer 4T1 cells as a model cell, its photo / dark toxicity was studied.

[0073] 4T1 cells were fed at a rate of 1×10 4 After seeding cells into 96-well plates and culturing for 24 hours, DMEM complete medium containing NTYOL at concentrations of 0 μM, 0.5 μM, 1 μM, 2 μM, 4 μM, and 8 μM was added for further culturing. The cells were then placed in a hypoxic environment (oxygen content below 0.1 g, carbon dioxide content 5%) or under normal culture conditions (oxygen content 21%, carbon dioxide concentration 5%) and incubated in a cell culture incubator for 12 hours. Finally, the 96-well plates were placed under a white light lamp (light power density 40 mW / cm²). -2 After 20 minutes of illumination, the cells were incubated in the original culture environment for another 12 hours. The supernatant was removed, and 100 mL of MTT solution was added for 2 hours of incubation. The culture was then terminated, the culture medium was aspirated from the wells, and 100 μL of dimethyl sulfoxide was added to each well. The wells were then shaken at low speed for 10 minutes to fully dissolve the crystals, and the OD value was measured using a microplate reader. Following the same procedure, the same phototoxicity and dark toxicity tests were performed on crystal violet (CV), a currently recognized oxygen-independent photodynamic type I photosensitizer.

[0074] Figure 5 (under normal oxygen conditions) and Figure 6 The graph (under hypoxic conditions) shows the toxicity evaluation results of NTYOL and CV on 4T1 cells. Under the same normal conditions, NTYOL exhibited lower dark toxicity and higher phototoxicity compared to CV. Furthermore, under the same hypoxic conditions, the photosensitizer CV almost lost its photodynamic effect due to the extreme lack of oxygen. In contrast, NTYOL still exhibited low dark toxicity under hypoxic conditions. Notably, after white light irradiation, the molecular biological activity was significantly enhanced, exhibiting high cytotoxicity, thus maintaining high phototoxicity under hypoxic conditions.

[0075] Example 7

[0076] Preparation of NTYOL nanoparticles (NTYOL NPs): Compound NTYOL (1 mg) and DSPE-PEG (1 mg) were dissolved in 1 mL of tetrahydrofuran at a mass ratio of 1:3 under ultrasonic dispersion, and then the uniformly mixed solution was quickly added to ultrapure water (10 mL). After further ultrasonic treatment for 3 min at 30% output (rated power 125 W) using a probe sonicator, the solution was purified by dialysis (molecular weight cut-off 3.5 kDa) for 2 days to remove the organic solvent and free small molecules, and then the nanoparticles were collected and concentrated by a centrifugal filter for further use. 2000 Preparation of NTYOL nanoparticles (NTYOL NPs): Compound NTYOL (1 mg) and DSPE-PEG (1 mg) were dissolved in 1 mL of tetrahydrofuran at a mass ratio of 1:3 under ultrasonic dispersion, and then the uniformly mixed solution was quickly added to ultrapure water (10 mL). After further ultrasonic treatment for 3 min at 30% output (rated power 125 W) using a probe sonicator, the solution was purified by dialysis (molecular weight cut-off 3.5 kDa) for 2 days to remove the organic solvent and free small molecules, and then the nanoparticles were collected and concentrated by a centrifugal filter for further use.

[0077] Example 8

[0078] Utilization of the light-induced biological activity of aromatic propargyl alcohol for in vivo tumor killing.

[0079] (1) Tumor model construction: Female BABL / c mice (5-6 weeks) were raised in a standard SPF laboratory animal room. 4T1 cells (1 x 10 6 cfu / mL) suspended in 100 μL of PBS were injected subcutaneously into the right dorsal side of each mouse to establish a mouse original tumor subcutaneous model. About 7 days later, when the tumor volume grew to about 100 mm 3 , the mice could be used for experiments.

[0080] (2) The model mice were randomly divided into four groups (n = 5), including: PBS group, PBS + L group, NTYOL NPs group and NTYOL NPs + L group, and different treatments were performed by tail vein administration (150 μL, the concentration of NTYOL NPs was 5 mg mL -1 ). After 36 h of administration, for the light irradiation group of mice, the tumor site of the mice was placed under white light for 30 min (power density 150 mW cm -2 ). The time of tail vein administration was recorded as day -1, and the time of light irradiation treatment was recorded as day 0. The body weight and tumor volume of the mice on the day of light irradiation treatment were recorded, and the body weight and tumor size of the mice were recorded and counted every 2 days thereafter. The results are shown in A and B of Figure 7 , the average tumor volume of the NTYOL NPs + L group was significantly lower than that of the other treatment groups, and no differences in body weight were observed in each group in the detection of body weight growth curves, indicating that no obvious physiological toxicity was observed in the mice after injection of the nanoparticles. This indicates that the nanoparticles can achieve tumor cell killing under light irradiation, thereby significantly inhibiting the growth of tumor cells, while also having good biological safety for normal cells.

[0081] The above embodiments of the present application are described in detail, it should be understood that the above described are only specific embodiments of the present application, and are not intended to limit the present application, any modification, supplement or similar way of substitution made within the principle range of the present application, should be included in the protection scope of the present application.

Claims

1. A method for photoinduced preparation of aromatic phenylpropynone, characterized in that, Under photoinduced conditions, the aromatic phenylpropynyl alcohol represented by formula (I) is converted into the aromatic phenylpropynone represented by formula (II). ； Formula (I); ； Formula (II); In formulas (I)-(II), R1 is a substituted phenyl group, and the substituent in the substituted phenyl group is triphenylvinyl.

2. The method according to claim 1, characterized in that, The conditions for photo-induced light are irradiation under ultraviolet or white light for ≥2s.

3. The method according to claim 1, characterized in that, The aromatic phenylpropynyl alcohol represented by formula (I) is prepared by the following method: (1) Compound 5-Bromo-2-nitrobenzaldehyde was heated to 60-80°C under an inert atmosphere and subjected to the Suzuki reaction for 6-24 hours. After the reaction was completed, the nitrobenzaldehyde derivative was obtained by post-treatment. R1 is a substituted phenyl group, and the substituent in the substituted phenyl group is triphenylvinyl. (2) Prepare an organic solution of nitrobenzaldehyde derivative, add Grignard reagent under ice bath conditions, react for 2-6 hours under an inert gas atmosphere, and after the reaction is completed, obtain aromatic phenylpropynyl alcohol as shown in formula (I) through post-treatment.

4. A compound, characterized in that, The compound is an aromatic phenylpropynol or an aromatic phenylpropynone. The structural formula of the aromatic phenylpropynol is shown in formula (I), and the structural formula of the aromatic phenylpropynone is shown in formula (II). R1 ​​is a substituted phenyl group, and the substituent in the substituted phenyl group is a triphenylvinyl group. ； Formula (I); ； Formula (II).

5. The use of a compound in the preparation of a tumor therapeutic drug, characterized in that, The tumor is breast cancer; the structure of the compound is shown in formula (I), where R1 is a substituted phenyl group and the substituent in the substituted phenyl group is triphenylvinyl. ； Formula (I).

6. The application according to claim 5, characterized in that, The tumor treatment drug comprises nanoparticles, and the nanoparticles comprise aromatic phenylpropynyl alcohol as shown in formula (I).

7. The application according to claim 6, characterized in that, The nanoparticles were prepared by mixing aromatic phenylpropynyl alcohol as shown in formula (I) with a surfactant and co-precipitating.

8. The use of a compound in the preparation of fluorescent labeling reagents for peptides or proteins, characterized in that, The structure of the compound is shown in formula (I), where R1 is a substituted phenyl group and the substituent in the substituted phenyl group is triphenylvinyl. ； Formula (I).

9. The application according to claim 8, characterized in that, The aromatic phenylpropynyl alcohol shown in formula (I) is brought into contact with polypeptides or proteins.

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