Hemicyanine compound as well as preparation method and application thereof

Abstract

The invention discloses a hemicyanine compound and a preparation method and application thereof.The hemicyanine compound comprises an anion X-and a cation shown in the following structure: # imgabs0. The hemicyanine compound IITC of the scheme structure can start a self-destruction mechanism under the long-time illumination effect, and therefore potential phototoxic reaction is effectively avoided; due to the characteristic, the photosensitizer becomes a potential self-destruction type organic photosensitizer candidate, and a new application way is opened up in a treatment strategy for resisting triple negative breast cancer (TNBC). The hemicyanine compound shows an excellent photocatalytic treatment effect on highly invasive triple-negative breast cancer cell lines (such as 4T1 cells), and the mechanism of the hemicyanine compound is that the hemicyanine compound is activated by illumination with a specific wavelength, so that the growth inhibition effect on target cancer cells is remarkably enhanced.

Description

Technical Field

[0001] The present invention relates to the technical field related to medicinal chemistry, and in particular to a hemicyanine compound and a preparation method and application thereof. Background Art

[0002] Breast cancer is one of the malignant tumors, and the optimization of its treatment plan has always been the focus of medical attention. Among them, triple-negative breast cancer (TNBC) is a highly invasive and poorly prognostic subtype of breast cancer. It is characterized by the lack of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). Due to the lack of specific targets for TNBC, traditional hormone therapy and targeted therapy have limited efficacy and currently rely mainly on chemotherapy. However, chemotherapy is often accompanied by serious toxic side effects and drug resistance problems. Therefore, there is an urgent need to develop new treatment strategies.

[0003] Photodynamic therapy (PDT) is an emerging cancer treatment method that provides a new approach to the treatment of TNBC due to its advantages such as strong selectivity, minimal trauma, and few side effects. PDT relies on photosensitizers to produce reactive oxygen species (such as singlet oxygen) under specific wavelengths of light to induce apoptosis or necrosis of tumor cells. However, at present, the clinical application of photosensitizers is still limited by some factors, such as the slow metabolism of photosensitizers after treatment, which may remain in the body for a long time, thereby inducing phototoxic reactions and affecting the patient's quality of life. Therefore, the development of self-destructive photosensitizers that can automatically degrade or inactivate after the completion of treatment has become a research hotspot in the field of PDT.

[0004] Zwitterionic hemicyanine dyes, with their good water solubility, biocompatibility and cell penetration, show unique advantages in PDT applications. Through precise design, the self-destruction mechanism is integrated into the structure of hemicyanine dyes, which can achieve rapid inactivation or degradation of photosensitizers after light exposure, thereby significantly reducing the risk of phototoxicity and improving the safety of treatment.

[0005] The use of 4T1 cell line as an in vitro research model for TNBC is highly representative and practical. The highly invasive and metastatic properties of this cell line enable it to simulate the complex biological behavior of TNBC in vivo. By testing the photodynamic therapy effect of self-destructive hemicyanine dyes in this cell line, its anti-tumor activity, metastasis inhibition ability and safety can be systematically evaluated. The photodynamic effect of the dye under specific lighting conditions was evaluated by cytotoxicity experiments and cell apoptosis / necrosis detection; and the therapeutic effect and safety of the dye on TNBC in vivo were further verified by using small animal models (such as BALB / c mice). The results of the study will reveal the high efficiency and safety of self-destructive hemicyanine dyes in PDT, providing a new, safer and more effective treatment option for TNBC patients. In addition, the success of this study may also promote the application of PDT technology in the treatment of other types of malignant tumors and promote overall progress in the field of cancer treatment.

[0006] Therefore, it is of great significance to design a hemicyanine compound that has a photodynamic effect on 4T1 cells and can be self-destructive. Summary of the invention

[0007] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention provides a hemicyanine compound, which has a strong photocatalytic therapeutic effect on 4T1 cells and is self-destructive.

[0008] The invention also provides a method for preparing the compound.

[0009] The present invention also provides the application of the above compounds.

[0010] According to one aspect of the present invention, a hemicyanine compound (IITC) is provided, wherein the compound comprises an anion X - And the cation shown in the following structure:

[0011]

[0012] According to a preferred embodiment of the present invention, at least the following beneficial effects are achieved: the semi-cyanine compounds of the present invention can initiate a self-destruction mechanism under long-term illumination, thereby effectively avoiding potential phototoxic reactions. This property makes them highly potential candidates for self-destructive organic photosensitizers, opening up new application avenues in the treatment strategy against triple-negative breast cancer (TNBC). The semi-cyanine compounds exhibit excellent photocatalytic therapeutic effects against highly invasive triple-negative breast cancer cell lines (such as 4T1 cells), and the mechanism is that they are activated by illumination of a specific wavelength, significantly enhancing the growth inhibition of target cancer cells.

[0013] Specifically, under appropriate lighting conditions, the compound showed extremely high sensitivity to 4T1 cells, with a half inhibitory concentration (IC50) as low as 0.081μM, indicating that it can effectively inhibit the proliferation of cancer cells at extremely low concentrations. This property is of great significance for reducing drug dosage and reducing treatment side effects. Under dark conditions without light, the cytotoxicity of the compound was significantly reduced, and the IC50 value was increased to 18.07μM. This difference not only ensures the safety of treatment, but also highlights the key role of light in activating its therapeutic activity.

[0014] Furthermore, the calculated phototherapy index (PI) was as high as 223, which is an important indicator to measure the balance between the therapeutic efficiency and safety of photosensitizers. A high PI value means that the compound has extremely high therapeutic efficiency under light while maintaining good safety, providing a strong scientific basis for the photodynamic therapy of triple-negative breast cancer.

[0015] Therefore, the semicyanine compounds of the present invention not only show unique advantages as self-destructive photosensitizers, but their experimental results in the field of triple-negative breast cancer treatment also indicate that they have important research value and broad application prospects in the future development of efficient and low-toxic photodynamic therapy strategies.

[0016] In some embodiments of the present invention, the X - Cl - Br - ,I - or PF 6 - , preferably PF 6 - .

[0017] According to another aspect of the present invention, a method for preparing the above-mentioned hemicyanine compound is provided, comprising the following steps:

[0018] S1. react 5-iodo-2,3,3-trimethyl-3H-indole with 6-bromohexyltrimethylammonium bromide, and the product obtained after the reaction is reacted with an anion X - Compounds for ion exchange;

[0019] S2. reacting the compound obtained in step S1 with 3,4-dihydroxy-3-cyclobutene-1,2-dione and quinoline to obtain the hemicyanine compound.

[0020] The preparation method according to a preferred embodiment of the present invention has at least the following beneficial effects: the preparation method of the present invention is easy to operate and has good industrial application prospects.

[0021] In some embodiments of the present invention, the step S1 specifically comprises mixing 5-iodo-2,3,3-trimethyl-3H-indole and 6-bromohexyltrimethylammonium bromide and adding solvent I, and reacting under heating.

[0022] In some embodiments of the present invention, in step S1, the conditions for the reaction under heating include at least one of the following conditions:

[0023] 1) The temperature is 85-100°C;

[0024] 2) Duration is 3-7 days;

[0025] 3) The heating method is heating reflux.

[0026] In some embodiments of the present invention, in step S1, the conditions for the reaction under heating include at least one of the following conditions:

[0027] 1) The temperature is 90°C;

[0028] 2) The duration is 7 days;

[0029] 3) The heating method is heating reflux.

[0030] In some embodiments of the present invention, the molar ratio of 5-iodo-2,3,3-trimethyl-3H-indole to 6-bromohexyltrimethylammonium bromide is 1 to 1.3:1.

[0031] In some embodiments of the present invention, the molar ratio of 5-iodo-2,3,3-trimethyl-3H-indole to 6-bromohexyltrimethylammonium bromide is 1.2:1.

[0032] In some embodiments of the present invention, 5-iodo-2,3,3-trimethyl-3H-indole and 6-bromohexyltrimethylammonium bromide are added with solvent I and PF 6 - NH 4 PF 6 , then reflux at 90°C for 7d, cool the reactant to room temperature, and remove solvent I. The reaction formula of this step is as follows:

[0033]

[0034] In some embodiments of the present invention, the solvent I is acetonitrile.

[0035] In some embodiments of the present invention, in step S2, the compound obtained in step S1 is mixed with 3,4-dihydroxy-3-cyclobutene-1,2-dione and quinoline after ion exchange and solvent II is added, and the mixture is reacted under heating to obtain the semi-cyanine self-destructive organic photosensitizer IITC.

[0036] In some embodiments of the present invention, the solvent II is selected from a mixture of n-butanol / toluene or a mixture of n-butanol / benzene (preferably, the volume ratio is 1:1).

[0037] In some embodiments of the present invention, in step S2, the conditions for the reaction under heating include at least one of the following conditions:

[0038] 1) The temperature is 105-115°C;

[0039] 2) The duration is 24 to 48 hours;

[0040] 3) The heating method is heating reflux.

[0041] In some embodiments of the present invention, the conditions for the reaction under heating include at least one of the following conditions:

[0042] 1) Temperature is 110°C;

[0043] 2) The time is 24 hours;

[0044] 3) The heating method is heating reflux.

[0045] In some embodiments of the present invention, in step S2, the molar ratio of the compound prepared in step S1 to 3,4-dihydroxy-3-cyclobutene-1,2-dione and quinoline is 2 to 2.3:1:10.

[0046] In some embodiments of the present invention, in step S2, the molar ratio of the compound prepared in step S1 to 3,4-dihydroxy-3-cyclobutene-1,2-dione and quinoline is 2.2:1:10.

[0047] In some embodiments of the present invention, in step S2, the compound prepared in step S1 is mixed with 3,4-dihydroxy-3-cyclobutene-1,2-dione and a mixture of n-butanol / toluene (1:1) is added, and then refluxed at 110° C. for 24 hours, the reactants are cooled to room temperature, the solvent is dried, acetone and ethyl acetate solution are added to precipitate solids, and after dissolution, methanol and ethyl acetate solution are added to precipitate solids. The reaction formula of this step is as follows:

[0048]

[0049] According to another aspect of the present invention, the use of the above compound in the preparation of anti-tumor drugs is proposed.

[0050] According to the application of a preferred embodiment of the present invention, at least the following beneficial effects are achieved: the compound of the scheme of the present invention has good water solubility, and can generate singlet oxygen ions and reactive oxygen species under light irradiation, and the excessive accumulation of singlet oxygen ions and reactive oxygen species in the body is closely related to oxidative stress, and therefore, it can kill tumor cells or inhibit their growth by inducing oxidative stress. The compound of the scheme of the present invention has a strong ability to generate superoxide anions after light irradiation, and can cause oxidative stress in tumor cells, and therefore, it has good anti-tumor potential, thereby indicating that it has good application prospects in the field of anti-tumor drugs.

[0051] In some embodiments of the present invention, the anti-tumor drug is an anti-triple-negative breast cancer drug. The compound of the present invention has a strong inhibitory effect on 4T1 cell proliferation and has a good application prospect in the anti-triple-negative breast cancer.

[0052] In some embodiments of the present invention, the triple-negative breast cancer is a triple-negative breast cancer cell line (4T1 cells) induced in BALB / c mice. The compounds of the present invention have good application prospects in the preparation of drugs against triple-negative breast cancer.

[0053] In some embodiments of the present invention, the drug further comprises a pharmaceutically acceptable carrier and / or excipient, that is, the drug or photosensitizer as the main active ingredient is mixed with a pharmaceutically acceptable carrier and / or excipient to prepare a composition, and is prepared into a clinically acceptable dosage form.

[0054] In some embodiments of the present invention, the excipient refers to diluents, binders, lubricants, disintegrants, solubilizers, stabilizers and other pharmaceutical matrices that can be used in the pharmaceutical field.

[0055] In some embodiments of the present invention, different pharmaceutical excipients used in pharmaceutical dosage forms may vary depending on the specific medical application. Pharmaceutical excipients can be used to adjust the solubility and bioavailability of the drug, increase its stability, adjust the host's immunogenic response and act as an emulsifier, antioxidant, aerosol propellant, tablet binder, tablet disintegrant. Preferred pharmaceutical excipients include but are not limited to diluents, cosolvents, stabilizers, adhesives, fillers, coating agents, disintegrants, lubricants and sweeteners that are compatible with the use of photosensitizers. Other pharmaceutical bases may also be included.

[0056] In some embodiments of the present invention, the carrier is an acceptable functional pharmaceutical excipient in the pharmaceutical field, including surfactants, suspending agents, emulsifiers and some new pharmaceutical polymer materials, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid, etc.

[0057] In some embodiments of the present invention, the present invention has no particular restrictions on the dosage form of the above-mentioned drugs, and the active substance can be administered together with an assimilable edible carrier, an inert diluent, or directly combined with food. Drug dosage forms include, but are not limited to, hard-shell or soft-shell gelatin capsules, tablets, pills, powder injections, solutions, suspensions, elixirs, syrups, wafers, gels, buccal or sublingual tablets, films, suppositories, and enemas.

[0058] In other embodiments of the present invention, the drug of the present invention can be prepared without any formulation adjuvant or using other drug delivery systems known in the prior art, such as liposome-forming components, vectored and non-vectored proteins, organic and inorganic nanoparticles, nanoemulsions and microemulsions, nanocrystals, a single solvent or a suitable solvent mixture, components such as lactose, polyvinyl pyrrolidone (PVP), etc.

[0059] In some embodiments of the present invention, the prepared drug can be administered orally, involving any part of the gastrointestinal tract (such as mouth, pharynx, esophagus, stomach, small intestine (duodenum, jejunum, ileum), large intestine (cecum, colon, rectum) and anus) or parenterally (such as intravenous, subcutaneous, intraperitoneal or local). If certain drugs are unstable under gastric conditions, they can be prepared into enteric-coated tablets.

[0060] According to another aspect of the present invention, an anti-tumor organic photosensitizer is provided, wherein the active ingredient of the anti-tumor metal photosensitizer includes the above-mentioned hemicyanine compound.

[0061] The photosensitizer is the main active ingredient, and other active ingredients may also be added.

[0062] According to another aspect of the present invention, a self-destruction method for the above-mentioned hemicyanine compounds is provided, comprising the following steps: irradiating with light having a wavelength of 630 to 640 nm (preferably 635 nm) for more than 2 hours.

[0063] Self-destruction can be achieved through a certain period of light exposure, which significantly reduces the risk of phototoxicity and improves the safety of treatment.

[0064] Other features and advantages of the present invention will be set forth in the description which follows, and in part will be apparent from the description, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0066] Figure 1is the H NMR image of the hemicyanine compound prepared in the embodiment of the present invention;

[0067] Figure 2 The ultraviolet absorption spectrum of the hemicyanine compound prepared in the embodiment of the present invention;

[0068] Figure 3 This is a graph showing the change in the ultraviolet absorption spectrum of the hemicyanine compound prepared in an embodiment of the present invention under light in water;

[0069] Figure 4 This is a graph showing the change in the ultraviolet absorption spectrum of the hemicyanine compound obtained in an embodiment of the present invention under illumination in a cell culture medium;

[0070] Figure 5 This is a NAPH test result diagram of the photocatalytic oxidation of hemicyanine compounds prepared in an embodiment of the present invention;

[0071] Figure 6 This is a NAPDH test result diagram of the photocatalytic oxidation of hemicyanine compounds obtained in an embodiment of the present invention;

[0072] Figure 7 This is a graph showing the test results of the ability of the hemicyanine compounds prepared in the examples of the present invention to generate superoxide anions;

[0073] Figure 8 This is a graph showing the test results of the ability of the hemicyanine compounds prepared in the examples of the present invention to generate singlet oxygen;

[0074] Fig. 9 This is a graph showing the toxicity test results of the hemicyanine compounds prepared in an example of the present invention against triple-negative breast cancer (4T1 cells).

[0075] Fig.10 This is a graph showing the phototoxicity test results of the illumination product of the hemicyanine compound prepared in an embodiment of the present invention against triple-negative breast cancer (4T1 cells).

[0076] Fig.11 This is a graph showing the phototoxicity test results of the illumination product of the hemicyanine compound prepared in an embodiment of the present invention against triple-negative breast cancer (4T1 cells). DETAILED DESCRIPTION

[0077] The following will be combined with the embodiments to clearly and completely describe the concept of the present invention and the technical effects produced, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of them. Based on the embodiments of the present invention, other embodiments obtained by technicians in this field without creative work are all within the scope of protection of the present invention. The test methods used in the embodiments are conventional methods unless otherwise specified; the materials, reagents, etc. used, unless otherwise specified, can be reagents and materials obtained from commercial channels. Unless otherwise specified, the same parameter in each embodiment has the same value. The embodiments described below are exemplary and are only used to explain the present invention, and cannot be understood as limitations on the present invention.

[0078] In the description of the present invention, the description with reference to the terms "one embodiment", "some embodiments", "illustrative embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0079] The "room temperature" referred to in the present invention refers to 25±5°C, and is specifically 25°C in the embodiments.

[0080] Example

[0081] In this example, a hemicyanine compound was prepared, and its structure is shown in the formula:

[0082]

[0083] The specific preparation process is as follows:

[0084] S1. 5-iodo-2,3,3-trimethyl-3H-indole (0.342 g, 1.2 mmol) and 6-bromohexyltrimethylammonium bromide (0.303 g, 1 mmol) were mixed, 10 ml of acetonitrile was added, and the mixture was stirred at 90°C for 7 days. After the reaction was completed, the mixture was cooled to room temperature, the acetonitrile was dried by spin drying, methanol and ethyl acetate were added to precipitate solids, and methanol and saturated hexafluorophosphate aqueous solution were added to precipitate solids after filtration, and the solids were purified by aluminum peroxide column to obtain a purple solid powder (yield 22.18%).

[0085] The chemical reaction equation above is as follows:

[0086]

[0087] S2. The compound obtained in step S1 (57.47 mg, 80 μmol) was mixed with 3,4-dihydroxy-3-cyclobutene-1,2-dione (4.34 mg, 38.1 μmol) and quinoline (49.2 mg, 380.95 μmol) and a mixture of n-butanol / toluene (18 ml, v:v=1:1) was added, and then refluxed at 110°C for 24 h. The reactants were cooled to room temperature, the solvent was dried, acetone and ethyl acetate solution were added to precipitate solids, and after dissolution, methanol and ethyl acetate solution were added to precipitate blue-black solids. The yield was 42.3%.

[0088] The chemical reaction equation above is as follows:

[0089]

[0090] The solid powder obtained by the above operation was tested by nuclear magnetic resonance hydrogen spectrum. The test results are as follows: Figure 1 The high-resolution mass spectrum information of the compound is as follows.

[0091] 1HNMR (500 MHz, DMSO-d 6 )δ7.90 (s, 2H), 7.68 (d, J = 8.3Hz, 2H), 7.19 (d, J = 8.3Hz, 2H), 5.78 (s, 2H), 4.03 (d, J = 11.3Hz, 4H), 3.29 (s, 4H), 3.03 (s, 18H), 1.66 (s, 18H), 1.38 (dq, J=39.6, 7.6Hz, 10H).

[0092] In order to verify the application effect of the above photosensitizer, its performance was tested, as follows:

[0093] 1. Absorption spectrum measurement

[0094] The photosensitizer prepared by the above operation was used as a sample to prepare a 10 mM mother solution, which was then placed in solvents of different polarities (methanol (EtOH), N,N-dimethylformamide (DMF), water, acetonitrile (MeCN), dichloromethane (DCM) and dimethyl sulfoxide (DMSO)) to a final concentration of 10 μM. The absorbance of the solution was detected using an ultraviolet spectrophotometer. The results of the ultraviolet-visible absorption spectrum are shown in FIG. Figure 2 As shown in the figure, it can be seen that it has good light absorption ability in organic solvents. The maximum absorption wavelength in aqueous solution is 640nm), which may be because the strong hydrogen bonds and polar interactions of water molecules make the ground state and excited state energy difference of photosensitizer molecules larger. In N, N-dimethylformamide and dimethyl sulfoxide solutions, the two solutions have high polarity and strong polar interactions, making the excited state energy level of the photosensitizer most stable, resulting in a red shift in the absorption wavelength.

[0095] 2. Changes in the UV absorption spectrum of self-destructive photosensitizers in water and culture medium under illumination

[0096] The photosensitizer was dissolved to prepare a 10mM stock solution, which was then placed in pure water and cell culture medium (DMEM) to a final concentration of 10μM. The absorbance of the solution was detected using a UV spectrophotometer. The quartz cuvette was placed under 635nm for irradiation, and the cuvette was taken out at regular intervals to quickly measure the UV absorption spectrum and record the data. The results were as follows: Figure 3 (in aqueous solution) and Figure 4 (in DMEM cell culture medium).

[0097] from Figure 3 and Figure 4 It can be seen that under illumination with a 635nm light source, the ultraviolet peak absorption of the photosensitizer gradually decreases with time.

[0098] 3. Determination of the ability of photocatalytic oxidation of NADH

[0099] In the aqueous solution, the absorbance of the NADH absorption peak at 339nm was controlled to be 1, and this concentration was used for subsequent determination. The above compound was added to the above solution to make the concentration 10μM as the experimental group, and the reference was the same concentration compound solution without NADH. The light group was placed under 635nm irradiation, and the dark group was not irradiated. The UV-visible light spectrum of the solution was detected at regular intervals to observe the changes in UV absorption at 339nm. The results are as follows Figure 5 shown.

[0100] from Figure 5 It can be seen that under light, the absorption peak of the photosensitizer at 339nm decreases, showing a strong photocatalytic oxidation effect and good light selectivity.

[0101] 4. Determination of the ability of photocatalytic oxidation of NADPH

[0102] In the aqueous solution, the absorbance of the NADH absorption peak at 339nm was controlled to be 1, and this concentration was used for subsequent determination. The compound was added to the above solution to make the concentration of 10μM as the experimental group, and the reference was the same concentration of compound solution without NADPH. The light group was placed under 635nm irradiation, and the dark group was not irradiated. The UV-visible light spectrum of the solution was detected at regular intervals to observe the changes in UV absorption at 339nm. The results are as follows Figure 6 shown.

[0103] from Figure 6 It can be seen that under light, the absorption peak of the photosensitizer at 339nm decreases, showing a strong photocatalytic oxidation effect and good light selectivity.

[0104] 5. Determination of the ability to generate superoxide anions in solution

[0105] The superoxide anion probe dihydrorhodamine 123 (DHR123 for short) was used to measure the ability of the photosensitizer to generate superoxide anions. The change in fluorescence intensity of the sample to be tested and the mixed solution of DHR123 under different illumination times was monitored by a fluorescence spectrophotometer to reflect the ability to generate superoxide anions. At λex = 465nm, a mixed solution of photosensitizer (10μM) and DHR123 (10μM) was excited in a 1cm quartz test tube, and the blank control was a probe solution of the same concentration without adding compounds. The incident slit and the exit slit were set to 2.5nm. The superoxide anion generation ability was measured under 635nm illumination, and the test results are as follows: Figure 7 As shown in the figure, after illumination, the fluorescence of DHR123 was significantly enhanced, while the fluorescence enhancement of DHR123 in the blank control group and the dark group was not obvious, indicating that the photosensitizer has a strong ability to produce superoxide anions and has a certain light selectivity.

[0106] 6. Determination of the ability to generate singlet oxygen in solution

[0107] In order to detect the ability of the photosensitizer synthesized in the embodiment to generate singlet oxygen under illumination, the singlet oxygen probe 9,10-anthryl-bis(methylene)dimalonic acid (abbreviated as ABDA) was used to measure the ability of the photosensitizer to generate singlet oxygen. When singlet oxygen is generated in the solution, ABDA will immediately capture the singlet oxygen in the solution, react to generate an endogenous oxidation product, so that the characteristic absorption peak of ABDA decreases, wherein the rate of decrease of the ABDA absorption peak is the rate of generation of singlet oxygen. Thus, the change of the ultraviolet-visible absorption spectrum of the sample to be tested and the ABDA mixed solution under different illumination times can be monitored by an ultraviolet-visible spectrophotometer to reflect the ability to generate singlet oxygen.

[0108] Two aqueous solutions containing the same photosensitizer (10 μM) and ABDA reagent (100 μM) were placed in a cuvette and their singlet oxygen generation capacity was measured under 635 nm illumination. The test results are shown in Figure 8 As shown in the figure, after illumination, the characteristic absorption peak of ABDA at 378nm is significantly reduced, while the characteristic absorption peak of the dark group remains almost unchanged, indicating that the compound has a strong ability to produce singlet oxygen and is photoselective.

[0109] 7. Dark toxicity and phototoxicity test results

[0110] Resazurin solution is blue and is usually used as an acid-base indicator (orange at pH 3.8 to dark purple at pH 6.5) and a redox indicator. When performing cell activity tests, resazurin can penetrate into cells and be irreversibly reduced to pink by living cells, while red fluorescent resorufin appears. The absorbance or fluorescence intensity of resorufin is positively correlated with the number of cells and the reducing ability, so cell proliferation can be analyzed using an enzyme-linked immunofluorescence detector.

[0111] The steps of the Resazurin experiment are as follows:

[0112] (1) First, revive one tube of 4T1 tumor cells and culture them with fresh complete culture medium (DMEM medium + 10 vol% fetal bovine serum + 1 vol% penicillin-streptomycin mixture). After two passages, start the experiment.

[0113] (2) When the cells reached the logarithmic growth phase, they were seeded into two 96-well plates at a density of 6,000 cells / well (each well was cultured with 90 μL of culture medium, one plate was the light control group, and the other plate was the dark control group) and placed in a 37°C, 5% CO 2 Culture in an incubator.

[0114] (3) After the cells adhered to the wall, 10 μL of photosensitizer (100, 50, 10, 1, 0.1, 0.01 μM) of the corresponding concentration was added to each well, and the cells were gently shaken and placed in a carbon dioxide incubator (37°C, 5% CO 2 ) and incubate in the dark.

[0115] (4) After incubation for 6 h, the cell culture plates in the illumination group were placed under a 635 nm light source for 30 min (light dose of 44 J / cm 2 ), and then returned to the incubator for continued incubation in the dark for 24 h (the cells in the dark control group were kept in the incubator in the dark for incubation).

[0116] (5) After incubation for 24 h, the culture medium was discarded from each well, and 80 μL of resazurin (100 mg / mL) was added to each well. After further incubation in a 37°C incubator for 2.5 to 4 h, EX540 / EM590 was detected using the fluorescence plate of an enzyme-linked immunosorbent assay (ELISA) to calculate the cell proliferation inhibition rate and obtain the IC 50 value (the concentration of compound at which the inhibition rate is equal to 50%).

[0117] Test results such as Fig. 9 As shown in the figure, it can be seen that the resazurin method detects the killing effect of different concentrations of photosensitizer on triple-negative breast cancer cell line (4T1 cells) under dark and light treatment conditions. In the absence of light, the IC 50The IC value of the triple-negative breast cancer cell line (4T1 cells) under light conditions was 18.07 μM. 50 The photocatalytic activity of the photosensitizer is 0.081 μM, and the phototherapy index PI is as high as 223, indicating that the photosensitizer of the present invention has a strong photocatalytic therapeutic effect.

[0118] 8. Toxicity test results of light-induced products against triple-negative breast cancer (4T1 cells)

[0119] (1) When 4T1 cells reached the logarithmic growth phase, they were seeded into two 96-well plates at a density of 6,000 cells / well (each well was cultured with 90 μL of culture medium, one plate was the light control group, and the other plate was the dark control group). 2 Culture in an incubator until the cells adhere.

[0120] (2) Photosensitizers of corresponding concentrations and commercially available photosensitizers Ce6 and verteporfin (100, 50, 10, 1, 0.1, and 0.01 μM) were placed under a 635 nm light source for 2 h to generate corresponding illumination products.

[0121] (3) After the cells adhered to the wall, 10 μL of the corresponding concentration of illumination product was added to each well, and the mixture was gently shaken and placed in a carbon dioxide incubator (37°C, 5% CO). 2 ) and incubate in the dark.

[0122] (4) After incubation for 6 h, the cell culture plates in the illumination group were placed under a 635 nm light source for 30 min (light dose of 44 J / cm 2 ), and then returned to the incubator for continued incubation in the dark for 24 h (the cells in the dark control group were kept in the incubator in the dark for incubation).

[0123] (5) After incubation for 24 h, the culture medium was discarded from each well, and 80 μL of resazurin (100 mg / mL) was added to each well. After further incubation in a 37°C incubator for 2.5 to 4 h, EX540 / EM590 was detected using the fluorescence plate of an enzyme-linked immunosorbent assay (ELISA) to calculate the cell proliferation inhibition rate and obtain the IC 50 value (the concentration of compound at which the inhibition rate is equal to 50%).

[0124] Test results such as Fig.10 (Light) and Fig.11(dark). As can be seen from the figure, the photodegradation products of the three photosensitizers after being exposed to light for the same time are not toxic when placed in the dark, and the products of the semi-cyanine photosensitizer after being exposed to light are almost non-cytotoxic, which indicates that the compound can be automatically degraded or lose activity after being exposed to light, and compared with Ce6 and verteporfin, the semi-cyanine photosensitizer has a faster metabolic rate, which helps to reduce the residence time of the photosensitizer in the body, thereby reducing the accumulation of toxic effects and reducing damage to normal cells or tissues.

[0125] In summary, the structural compounds of the scheme of the present invention have a good inhibitory effect against triple-negative breast cancer cells under light, and can self-destruct after prolonging the continuous light exposure time, thereby reducing phototoxicity and improving the safety of medication. They can be prepared into drugs such as anti-tumor photosensitizers, and have good application prospects in the field of photodynamic therapy for tumors.

[0126] The embodiments of the present invention are described in detail above, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge scope of ordinary technicians in the relevant technical field without departing from the purpose of the present invention.

Claims

1. A hemicyanine compound, characterized in that: The compound includes an anion X - And the cation shown in the following structure:

2. The hemicyanine compound according to claim 1, characterized in that: The X - Cl - Br - ,I - or PF6 - , preferably PF6 - .

3. The method for preparing a hemicyanine compound according to claim 1 or 2, characterized in that: The steps include: S1. react 5-iodo-2,3,3-trimethyl-3H-indole with 6-bromohexyltrimethylammonium bromide, and the product obtained after the reaction is reacted with an anion X - Compounds for ion exchange; S2. reacting the compound obtained in step S1 with 3,4-dihydroxy-3-cyclobutene-1,2-dione and quinoline to obtain the hemicyanine compound.

4. The preparation method according to claim 3, characterized in that: The step S1 specifically comprises mixing 5-iodo-2,3,3-trimethyl-3H-indole with 6-bromohexyltrimethylammonium bromide and adding solvent I, and the reaction conditions under heating include at least one of the following conditions: 1) The temperature is 85-100°C; 2) Duration is 3-7 days; 3) The heating method is heating reflux; 4) The solvent I is acetonitrile.

5. The preparation method according to claim 3, characterized in that: In the step S2, the compound obtained in step S1 is mixed with 3,4-dihydroxy-3-cyclobutene-1,2-dione and quinoline after ion exchange, and solvent II is added, and the mixture is reacted under heating to obtain the semi-cyanine self-destructive organic photosensitizer. The conditions for the reaction under heating include at least one of the following conditions: 1) The temperature is 105-115°C; 2) The duration is 24 to 48 hours; 3) The heating method is heating reflux; 4) The solvent II is selected from a mixture of n-butanol / toluene or a mixture of n-butanol / benzene.

6. Use of the hemicyanine compound according to claim 1 or 2 in the preparation of anti-tumor drugs.

7. The use according to claim 6, characterized in that: The anti-tumor drug is an anti-triple-negative breast cancer drug; preferably, the triple-negative breast cancer is a triple-negative breast cancer cell line induced in BALB / c mice.

8. The use according to claim 6, characterized in that: The medicament further comprises a pharmaceutically acceptable carrier and / or excipient.

9. An anti-tumor organic photosensitizer, characterized in that: The active ingredient of the anti-tumor metal photosensitizer includes the hemicyanine compound as claimed in claim 1 or 2.

10. The self-destruction method of hemicyanine compounds according to claim 1 or 2, characterized in that: The method comprises the following steps: irradiating with light having a wavelength of 630 to 640 nm for more than 2 hours.

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