Selenium-containing rhodamine derivative and application thereof

Abstract

The invention relates to the technical field of rhodamine derivatives, and particularly discloses a selenium-containing rhodamine derivative and application thereof. The rhodamine derivative containing selenium heavy atoms is innovatively provided, and the visible light absorption wavelength of the rhodamine derivative is close to a near-infrared region of 700 NM, so that the rhodamine derivative can penetrate through deeper tissue parts for deep treatment; meanwhile, the rhodamine derivative containing the selenium heavy atoms also shows a high triplet state conversion rate, and can efficiently generate active oxygen species under an illumination excitation condition, so that the killing effect on cancer cells and cancer tissues is enhanced. Compared with the prior art, the rhodamine derivative containing the selenium heavy atoms can realize efficient removal of cancer cells and cancer tissues by only needing nanomole-level concentration, so that the treatment efficiency is remarkably improved, and the drug use dosage is reduced; meanwhile, the preparation process of the rhodamine derivative is simple and easy to implement, high synthesis yield can be obtained, and the rhodamine derivative is more suitable for large-scale and low-cost production and application.

Description

Technical Field

[0001] The present application relates to the technical field of rhodamine derivatives, and in particular to a selenium-containing rhodamine derivative and its application. Background Art

[0002] Photodynamic therapy has become one of the important medical technologies for treating malignant tumors due to its minimally invasive nature, high biocompatibility, and high spatiotemporal selectivity.

[0003] Photodynamic therapy refers to the intravenous injection of drugs to selectively accumulate in tumor tissues. After a specific period of time, the tumor tissues are irradiated with light of a specific wavelength. At this time, the photosensitizers enriched in the tumor tissues will trigger a series of photophysical and chemical reactions under the excitation of light, thereby producing cytotoxic reactive oxygen species, thereby killing cancer cells and destroying tumor tissues. Currently, a variety of photosensitizers have been applied to cancer treatment research. For example, rhodamine has been widely used as a new type of photosensitizer for monitoring and diagnosis during photodynamic therapy. In biological imaging, the distribution of photosensitizers in cells or tissues can be clearly observed, helping doctors to accurately determine the location of the lesion.

[0004] However, the absorption and emission wavelengths of rhodamine are in the visible light region. Due to the weak penetration of wavelengths in this region, rhodamine is difficult to fully exert its effect when treating deeper tissue sites. Existing technologies usually expand the conjugated system within the molecule to promote more effective delocalization and transfer of electrons within the molecule, causing the absorption spectrum to red-shift, and enhancing the absorption width of rhodamine in the near-infrared band so that rhodamine can act on deeper tissue sites. However, the increase in the conjugated system may cause intermolecular aggregation, which will reduce the light absorption capacity of the aggregated photosensitizer molecules, thereby reducing the yield of reactive oxygen species such as singlet oxygen, and weakening the ability to kill cancer cells. Summary of the invention

[0005] In view of this, the purpose of the present application is to provide a selenium-containing rhodamine derivative and its application, which is used to solve the problems of narrow absorption width of the existing rhodamine in the near-infrared band and low yield of active oxygen species.

[0006] In order to achieve the above technical purpose, the present application provides a selenium-containing rhodamine derivative, the chemical structure of which is as follows:

[0007] .

[0008] Further, the R1 substituent is one of the following groups,

[0009] .

[0010] Further, the R2 substituent is one of the following groups,

[0011] .

[0012] Further, the R3 substituent is one of the following groups,

[0013] .

[0014] Further, the R4 substituent is one of the following groups,

[0015] .

[0016] Further, the R5 substituent is one of the following groups,

[0017] .

[0018] Further, R1 is selected from 1,2,3,4-tetrahydroquinoline or N,N-diethylaniline, R2 is selected from ethyl, R3 is selected from benzene ring, R4 is selected from ester group, and R5 is selected from hydrogen atom.

[0019] Further, the chemical structural formula is one of the following,

[0020] .

[0021] The present application provides a method for preparing a rhodamine derivative A, comprising the following steps:

[0022] Step S1, under an inert atmosphere, reacting 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-carboxylic acid)benzoic acid to obtain 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one;

[0023] Step S2, dissolving 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one in a solvent, adding phosphorus oxychloride, heating under reflux, and obtaining a selenium-containing rhodamine derivative.

[0024] Furthermore, in step S1, the reaction temperature is 50-80°C, the reaction time is 8-14 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-formyl)benzoic acid is 1:(1-3); in step S2, the reaction temperature is 0-50°C, the reaction time is 4-6 hours, and the molar ratio of 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one and phosphorus oxychloride is 1:(2-3).

[0025] The present application provides a method for preparing a rhodamine derivative B, comprising the following steps:

[0026] Step S1, under an inert atmosphere, reacting 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid to obtain 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one;

[0027] Step S2, dissolving 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one in a solvent, adding phosphorus oxychloride, and heating under reflux to obtain a selenium-containing rhodamine derivative B.

[0028] Further, in step S1, the reaction temperature is 60-80°C, the reaction time is 10-12 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenazine to 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid is 1:(1-3);

[0029] In the step S2, the reaction temperature is 0-50° C., the reaction time is 4-10 hours, and the molar ratio of 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one to phosphorus oxychloride is 1:(2-3).

[0030] The present application provides an application of a rhodamine derivative for treating malignant tumors.

[0031] Furthermore, the malignant tumor is one or more of human breast cancer cells, human non-small cell lung cancer cells, human glioma cells, human liver cancer cells, cervical cancer cells, and melanoma cells.

[0032] In summary, this application innovatively proposes a rhodamine derivative containing selenium heavy atoms, whose visible light absorption wavelength extends to more than 600NM, close to the near-infrared region of 700NM. This feature enables it to penetrate deeper tissue sites and is suitable for the treatment of malignant tumors in deep tissue sites; at the same time, the selenium heavy atoms in the rhodamine derivatives can enhance the intra-molecule intersystem crossing rate, so that the rhodamine derivatives show a high triplet conversion rate under the excitation conditions of light, and produce a large number of reactive oxygen species to enhance the killing effect on cancer cells and cancer tissues. Compared with the prior art, the rhodamine derivatives containing selenium heavy atoms in this application have excellent killing effects, and only nanomolar concentrations are required to achieve efficient removal of cancer cells and cancer tissues, significantly improving the treatment efficiency and reducing the dosage of drugs.

[0033] The method for preparing rhodamine derivatives provided in this application presents significant advantages. Its synthetic route is simple, and only two main synthetic steps are required to successfully prepare the target product. Compared with traditional complex synthetic techniques, this method not only has a simple synthetic process and is easy to implement, but also can obtain a higher synthetic yield. This is not only conducive to the rapid and low-cost preparation of the derivatives on a laboratory scale, but also lays a solid foundation for subsequent large-scale industrial production, and has broad application prospects and potential value in the field of drug research and development and production. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0035] Figure 1 A schematic diagram of the chemical synthesis of the selenium-containing rhodamine derivative A provided in Example 1 of the present application;

[0036] Figure 2 The hydrogen nuclear magnetic resonance spectrum of 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one provided in Example 1 of the present application;

[0037] Figure 3 This is the hydrogen nuclear magnetic resonance spectrum of the selenium-containing rhodamine derivative A provided in Example 1 of the present application;

[0038] Figure 4 A schematic diagram of the chemical synthesis of the selenium-containing rhodamine derivative B provided in Example 2 of the present application;

[0039] Figure 5 The hydrogen nuclear magnetic resonance spectrum of 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one provided in Example 2 of the present application;

[0040] Figure 6 This is the hydrogen nuclear magnetic resonance spectrum of the selenium-containing rhodamine derivative B provided in Example 2 of the present application;

[0041] Figure 7 Absorption spectra of selenium-containing rhodamine derivatives A and B provided in Examples 1 and 2 of the present application;

[0042] Figure 8 This is a graph showing the active oxygen generation capacity of the selenium-containing rhodamine derivatives provided in Examples 1 and 2 of the present application;

[0043] Fig. 9 This is a graph showing the growth inhibition of mouse breast cancer cells, human cervical cancer cells, and mouse melanoma cells by the selenium-containing rhodamine derivatives provided in Examples 1-2 of the present application. DETAILED DESCRIPTION

[0044] The technical solutions of the embodiments of the present application will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the specification of the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection requested by the present application.

[0045] The sources of all raw materials in the present invention are not particularly limited and can be purchased from the market or prepared according to conventional methods known to those skilled in the art.

[0046] The embodiment of the present invention provides a selenium-containing rhodamine derivative, the chemical structure of which is as follows:

[0047] .

[0048] It should be noted that the selenium-containing rhodamine derivatives proposed in this application are carefully designed based on the strategy of selenium heavy atoms to enhance the killing ability of photosensitizers on cancer cells. After the introduction of selenium heavy atoms in its chemical formula, the derivatives are endowed with unique optical and biological properties. From the perspective of optical properties, its visible light absorption wavelength is extended to the near-infrared band of 600NM to nearly 700NM, which gives it a significant advantage in the process of biological tissue penetration and can reach the lesion site more deeply. At the same time, the presence of selenium heavy atoms greatly improves the triplet conversion rate of selenium-containing rhodamine derivatives, so that under light excitation, oxygen species with strong oxidative activity can be produced with high efficiency. From the perspective of biological characteristics, the smaller radius of action and shorter lifespan of the active oxygen species themselves accurately define their scope of action. In the process of photodynamic therapy, only cancer cells that receive specific light irradiation are damaged, effectively avoiding unwarranted damage to surrounding normal cell tissues, thereby minimizing toxic side effects at the root, and significantly improving the accuracy and effectiveness of treatment. In addition, the good water solubility of selenium-containing rhodamine derivatives ensures their good dispersibility and transportability in the body. The cell membrane permeability enables them to smoothly cross the cell membrane and enter the cell to exert their effects, while the excellent biological tissue penetration provides a strong guarantee for their penetration into deep malignant tumor tissues.

[0049] In some embodiments, the R1 substituent is one of the following groups,

[0050] .

[0051] In some embodiments, the R2 substituent is one of the following groups,

[0052] .

[0053] In some embodiments, the R3 substituent is one of the following groups,

[0054] .

[0055] In some embodiments, the R4 substituent is one of the following groups,

[0056] .

[0057] In some embodiments, the R5 substituent is one of the following groups,

[0058] .

[0059] In some embodiments, R1 is selected from 1,2,3,4-tetrahydroquinoline or N,N-diethylaniline, R2 is selected from ethyl, R3 is selected from benzene ring, R4 is selected from ester group, and R5 is selected from hydrogen atom.

[0060] In some embodiments, the chemical formula is one of the following,

[0061] .

[0062] It should be noted that the selenium-containing rhodamine derivative SE-RHOD-A exhibits the strongest visible light absorption wavelength of 610NM. Under light conditions, the selenium-containing rhodamine derivative of formula A can have a significant killing effect on cells, especially cancer cells, and the killing effect is particularly prominent, which can effectively curb the invasion of cancer cells on normal human cells.

[0063] The embodiment of the present invention provides a method for preparing a rhodamine derivative A, comprising the following steps:

[0064] Step S1, under an inert atmosphere, 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-carbonyl)benzoic acid are reacted to obtain 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one.

[0065] Step S2, dissolving 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one in a solvent, adding phosphorus oxychloride, heating under reflux, and obtaining a selenium-containing rhodamine derivative A.

[0066] In some embodiments, in step S1, the reaction temperature is 50-80°C, the reaction time is 8-14 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-carbonyl)benzoic acid is 1:(1-3); in step S2, the reaction temperature is 0-50°C, the reaction time is 4-6 hours, and the molar ratio of 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one and phosphorus oxychloride is 1:(2-3).

[0067] The present invention provides a method for preparing a rhodamine derivative B, comprising the following steps:

[0068] Step S1, under an inert atmosphere, 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid are reacted to obtain 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one.

[0069] Step S2, dissolving 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one in a solvent, adding phosphorus oxychloride, and heating under reflux to obtain a selenium-containing rhodamine derivative B.

[0070] In some embodiments, in step S1, the reaction temperature is 60-80°C, the reaction time is 10-12 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenazine to 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid is 1:(1-3); in step S2, the reaction temperature is 0-50°C, the reaction time is 4-10 hours, and the molar ratio of 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one to phosphorus oxychloride is 1:(2-3).

[0071] The present application provides an application of a rhodamine derivative for treating malignant tumors.

[0072] In some embodiments, the malignant tumor is a human breast cancer cell, a human non-small cell lung cancer cell, a human glioma cell, a human liver cancer cell, a cervical cancer cell, or a melanoma cell.

[0073] The applicant further provides the following reference specific embodiments to describe the present invention. It should be noted that these embodiments are merely illustrative and do not limit the present invention in any way. Example 1

[0074] See also Figures 1 to 3 This embodiment provides a method for preparing the selenium-containing rhodamine derivative of formula A, comprising the following steps:

[0075] Step S1, preparation of 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one:

[0076] Argon was introduced into the chemical synthesis device for protection, and 10-ethyl-2-methoxy-10H-phenoselenazine (2g, 6.5mmOl), 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-formyl)benzoic acid (3g, 10mmOl) and 5ml methanesulfonic acid were added into the synthesis device, and then heated to 70°C and condensed and refluxed for 12 hours; after the reflux was completed, the heating was stopped and cooled to room temperature, and the pressure was -0.1MPA and the temperature was 4 The solvent was removed by reduced pressure distillation at 5°C to obtain an intermediate, which was purified by silica gel column chromatography (eluent: PE:EA=5:1) to obtain 1.5G of a blue solid with a yield of 48%. The blue solid was 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one, and its H NMR spectrum was as follows Figure 2 As shown;

[0077] Step S2, introduce argon into the chemical synthesis device for protection, take the 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one (1G, 1.8MMOL) prepared in step 1 and 10ML dichloromethane and place them in the chemical synthesis device, then add phosphorus oxychloride (600ΜL, 6.4MMOL) to the chemical synthesis device, heat to 50°C and then condense and reflux for 4H, stop heating and cool to room temperature after reflux, and press The solvent was removed by reduced pressure distillation at a pressure of -0.1 MPA and a temperature of 45°C to obtain an intermediate; 10 mL of dichloromethane, triethylamine (890 μL, 6.4 MM01) and methanol (260 μL, 6.4 MM01) were added to the intermediate, and stirring was continued. At the same time, TLC was used to monitor the reaction progress in real time. After the reaction, a crude product was obtained. The crude product was taken and the solvent was removed by reduced pressure distillation at a pressure of -0.1 MPA and a temperature of 45°C. Then, it was purified by recrystallization using methanol as a solvent to obtain 390 MG of a blue solid product A with a yield of 45%. The hydrogen nuclear magnetic resonance spectrum is shown as follows Figure 3 shown. Example 2

[0078] See also Figures 1 to 3 This embodiment provides a method for preparing the selenium-containing rhodamine derivative of formula B, comprising the following steps:

[0079] Step S1, preparation of 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one:

[0080] Argon gas was introduced into the chemical synthesis device for protection, and 10-ethyl-2-methoxy-10H-phenoselenazine (2 g, 6.5 mmol), 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid (3.1 g, 10 mmol), and 5 ml of methanesulfonic acid were added into the synthesis device, and then heated to 75°C and condensed and refluxed for 10 hours; after the reflux was completed, the heating was stopped and cooled to room temperature, and the solvent was removed by reduced pressure distillation under the conditions of -0.1 MPA and 45°C to obtain an intermediate, which was purified by silica gel column chromatography (eluent PE: EA = 3:1) to obtain 1.6 g of a blue solid with a yield of 43%; the blue solid was 3-(diethylamino)-7-ethyl-3'H, 7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one, and its hydrogen nuclear magnetic resonance spectrum was as shown in Figure 5 As shown;

[0081] Step S2, introduce argon gas into the chemical synthesis device for protection, take 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one (1G, 1.8MMOL) prepared in step 1 and 10ML dichloromethane and place them in the chemical synthesis device, then add phosphorus oxychloride (600ΜL, 6.4MMOL) into the chemical synthesis device, heat to 50°C and then condense and reflux for 4H, stop heating and cool to room temperature when reflux is completed, and then place the mixture under a pressure of -0.1MPA and a temperature of The solvent was removed by reduced pressure distillation at 45°C to obtain an intermediate; 10ML of dichloromethane, triethylamine (890μL, 6.4MMO1), and methanol (260μL, 6.4MMO1) were added to the intermediate, and stirring was continued. At the same time, TLC was used to monitor the reaction process in real time. After the reaction, a crude product was obtained. The crude product was taken and the solvent was removed by reduced pressure distillation at a pressure of -0.1MPA and a temperature of 45°C. Then, it was purified by recrystallization using methanol as a solvent to obtain 430MG of a blue solid product B with a yield of 45.5%. Its nuclear magnetic resonance hydrogen spectrum is shown as follows Figure 6 shown.

[0082] The selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B prepared in Examples 1 and 2 of the present application were subjected to performance tests to determine their effects on cell photodynamic therapy. The performance tests included spectral testing, reactive oxygen testing, and cell photodynamic therapy testing.

[0083] Detection 1: Spectrum test:

[0084] Selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B were dissolved in different solvents and tested by UV-visible spectrophotometer. The results are as follows: Figure 7As shown in the figure, it can be seen that the visible light absorption wavelength of the selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B involved in the present application is close to 700NM, and the highest absorption intensity is shown at 590NM and 610NM respectively. This shows that the absorption wavelength of the selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B has been red-shifted to the near-infrared region and can be applied to photodynamic therapy of deeper tissues.

[0085] Test 2: Active oxygen test:

[0086] The selenium-containing rhodamine derivative SE-RHOD-A or SE-RHOD-B is dissolved in DMSO solvent, and then 1,3-diphenylisobenzofuran (DPBF) is added to the solution to obtain a mixed solution. Subsequently, the mixed solution is irradiated with red light with a wavelength range of 620~650NM and a light intensity of 30MW / CM² for a certain period of time, and the mixed solution is tested using a UV-visible spectrophotometer, focusing on recording the absorbance values ​​of the mixed solution at a wavelength of 417NM under different illumination times, and drawing a curve of the absorbance value of 1,3-diphenylisobenzofuran changing with time, as shown in FIG. Figure 8 shown.

[0087] When active oxygen is generated, 1,3-diphenylisobenzofuran undergoes an irreversible oxidation reaction, and its absorption intensity at 417NM decreases rapidly. It can be clearly observed from the figure that the selenium-containing rhodamine derivatives SE-RHOD-A or SE-RHOD-B involved in the present application can quickly generate active oxygen under light conditions, indicating that the selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B have excellent photosensitization effects. When they are applied to fields such as photodynamic therapy, the rapidly generated active oxygen can effectively participate in the corresponding biochemical reactions and exert their therapeutic effects.

[0088] Test 3: Cell photodynamic therapy test:

[0089] Different cell lines were inoculated into 96-well plates at a cell density of 5000 cells / well. The 96-well plates were placed in a 37°C, CO 2 The cells were incubated in an incubator with a concentration of 5% CO for 24 hours. The cell lines included human breast cancer cells (4T1CELL), cervical cancer cells (HELA CELL), and melanoma cells (B16F10 CELL).

[0090] Then, the selenium rhodamine derivatives SE-RHOD-A and SE-RHOD-B were added to DMEM containing 10% fetal bovine serum to prepare mixed solutions of different concentrations. The mixed solutions of different concentrations were added to each well and placed in a cell culture incubator for incubation for 2 hours. The mixed solution in the well plate was removed with a pipette gun, replaced with a fresh mixed solution, and the well plate was exposed to red light with a wavelength range of 620~650NM and a light intensity of 30MW / CM² for 10 minutes. After the illumination, the well plate was placed in a cell culture incubator for further incubation for 22 hours, and then the mixed solution in the well plate was removed with a pipette gun, and 150UL of MTT culture medium with a concentration of 5MG / ML was added to the well plate. The well plate was moved to a cell culture incubator for incubation for 4 hours, the MTT culture medium in the plate wells was removed, and then 100UL of DMSO was added to fully dissolve the oxidation products of MTT. The absorbance of each well solution was accurately measured at a wavelength of 490 nm using an enzyme reader. The cell viability was calculated based on the linear relationship between the absorbance value and the number of cells, as shown in Tables 1 to 3. The cell viability data of SE-RHOD-A and SE-RHOD-B in different cell lines were fitted to calculate the IC of SE-RHOD-A or SE-RHOD-B. 50 The fitting results are as follows Fig. 9 shown.

[0091] Table 1

[0092]

[0093] According to Table 1 and Fig. 9 It can be seen from the data that when the addition concentration of SE-RNOD-A is 1.1 ΜM, the survival rate of 4T1 CELL is 9.65%; when the addition concentration of SE-RNOD-A is 10.0 ΜM, the survival rate of 4T1 CELL is 8.43%. When the addition concentration of SE-RNOD-B is 0.12 ΜM, the survival rate of 4T1 CELL is 13.69%; when the addition concentration of SE-RNOD-B is 10.0 ΜM, the survival rate of 4T1 CELL is 13.92%. When the addition amounts of SE-RNOD-A and SE-RNOD-B are 320 NM and 21 NM respectively, 50% of 4T1 CELL apoptosis can be achieved.

[0094] Table 2

[0095]

[0096] According to Table 2 and Fig. 9It can be seen from the data that when the addition concentration of SE-RNOD-A is 0.12 ΜM, the survival rate of HELA CELL is 19.69%; when the addition concentration of SE-RNOD-A is 1.1 ΜM, the survival rate of HELA CELL can reach a minimum of 12.67%. When the addition concentration of SE-RNOD-B is 0.12 ΜM, the survival rate of HELA CELL is 11.27%; when the addition concentration of SE-RNOD-B is 3.3 ΜM, the survival rate of HELA CELL is as low as 6.36%. When the addition amounts of SE-RNOD-A and SE-RNOD-B are 31 NM and 13 NM respectively, 50% of HELA CELL apoptosis can be achieved.

[0097] Table 3

[0098]

[0099] According to Table 3 and Fig. 9 It can be seen from the data that when the addition concentration of SE-RNOD-A is 0.22 ΜM, the survival rate of B16F10 CELL is 18.68%; when the addition concentration of SE-RNOD-A is 2.0 ΜM, the survival rate of B16F10 CELL is 13.75%. When the addition concentration of SE-RNOD-B is 0.22 ΜM, the survival rate of B16F10 CELL is 14.69%; when the addition concentration of SE-RNOD-B is 2.0 ΜM, the survival rate of B16F10 CELL is 11.59%. When the addition amounts of SE-RNOD-A and SE-RNOD-B are 55 NM and 52 NM respectively, 50% apoptosis of B16F10 CELL can be achieved.

[0100] In summary, the selenium-containing rhodamine derivatives provided in this application have significant killing effects on various cancer cells under light conditions. In the field of photodynamic therapy of malignant tumors, they demonstrate extremely considerable potential and application value, and open up a promising new photosensitizer path for tumor treatment.

[0101] The above are only preferred embodiments of the present application and are not intended to limit the present invention. Although the present application has been described in detail with reference to the examples, those skilled in the art can still modify the technical solutions recorded in the aforementioned examples or make equivalent substitutions for some of the technical features therein. However, any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A selenium-containing rhodamine derivative, characterized in that: The chemical structure is as follows: 。 2. The selenium-containing rhodamine derivative according to claim 1, characterized in that The R1 substituent is one of the following groups, 。 3. The selenium-containing rhodamine derivative according to claim 1, characterized in that The R2 substituent is one of the following groups, 。 4. The selenium-containing rhodamine derivative according to claim 1, characterized in that The R3 substituent is one of the following groups, 。 5. The selenium-containing rhodamine derivative according to claim 1, characterized in that: The R4 substituent is one of the following groups, 。 6. The selenium-containing rhodamine derivative according to claim 1, characterized in that The R5 substituent is one of the following groups, 。 7. The selenium-containing rhodamine derivative according to claim 1, characterized in that: R1 is selected from 1,2,3,4-tetrahydroquinoline or N,N-diethylaniline, R2 is selected from ethyl, R3 is selected from benzene ring, R4 is selected from ester group, and R5 is selected from hydrogen atom.

8. The selenium-containing rhodamine derivative according to claim 7, characterized in that: The chemical structural formula is one of the following, 。 9. A method for preparing the rhodamine derivative A as claimed in claim 8, characterized in that: The following steps are involved: Step S1, under an inert atmosphere, reacting 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-carboxylic acid)benzoic acid to obtain 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one; Step S2, dissolving 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenazine]-3-one in a solvent, adding phosphorus oxychloride, heating under reflux, and obtaining a selenium-containing rhodamine derivative A.

10. The method for preparing the rhodamine derivative A according to claim 9, characterized in that: In the step S1, the reaction temperature is 50-80° C., the reaction time is 8-14 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenazine to 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-formyl)benzoic acid is 1:(1-3); In the step S2, the reaction temperature is 0-50° C., the reaction time is 4-6 hours, and the molar ratio of 13'-ethyl-1', 2', 3', 4'-tetrahydro-3H, 13'H-spiro[isobenzofuran-1, 6'-pyrido[3', 2': 6, 7]chromeno[2, 3-B]phenoselenazine]-3-one to phosphorus oxychloride is 1:(2-3).

11. A method for preparing the rhodamine derivative B as claimed in claim 8, characterized in that: The following steps are involved: Step S1, under an inert atmosphere, reacting 10-ethyl-2-methoxy-10H-phenoselenazine and 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid to obtain 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one; Step S2, dissolving 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one in a solvent, adding phosphorus oxychloride, and heating under reflux to obtain a selenium-containing rhodamine derivative B.

12. The method for preparing the rhodamine derivative B according to claim 11, characterized in that: In the step S1, the reaction temperature is 60-80° C., the reaction time is 10-12 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenazine to 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid is 1:(1-3); In the step S2, the reaction temperature is 0-50° C., the reaction time is 4-10 hours, and the molar ratio of 13'-ethyl-1', 2', 3', 4'-tetrahydro-3H, 13'H-spiro[isobenzofuran-1, 6'-pyrido[3', 2': 6, 7]chromeno[2, 3-B]phenoselenazine]-3-one to phosphorus oxychloride is 1:(2-3).

13. A use of the rhodamine derivative according to any one of claims 1 to 8, characterized in that: Used to treat malignant tumors.

14. The use of the rhodamine derivative according to claim 13, characterized in that: The malignant tumor is one or more of human breast cancer cells, human non-small cell lung cancer cells, human glioma cells, human liver cancer cells, cervical cancer cells, and melanoma cells.

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