A selenium-containing rhodamine derivative and use thereof

Abstract

The application relates to the technical field of rhodamine derivatives, and specifically discloses a selenium-containing rhodamine derivative and application thereof. The application innovatively proposes a selenium-heavy-atom-containing rhodamine derivative, which has a visible light absorption wavelength close to the near-infrared region of 700 NM, which enables it to penetrate deeper tissue sites for deep treatment; meanwhile, the selenium-heavy-atom-containing rhodamine derivative also exhibits a high triplet state conversion rate, and can efficiently generate reactive oxygen species under light excitation conditions, thereby strengthening the killing effect on cancer cells and cancer tissues. Compared with the prior art, the selenium-heavy-atom-containing rhodamine derivative of the application can realize efficient removal of cancer cells and cancer tissues at a concentration of only nanomolar order, thereby significantly improving the treatment efficiency and reducing the drug dosage; meanwhile, the preparation process of the rhodamine derivative provided by the application is simple and easy to implement, and a relatively high synthesis yield can be obtained, and the rhodamine derivative is more suitable for large-scale, low-cost production and application.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of rhodamine derivatives, and particularly relates to a selenium-containing rhodamine derivative and application thereof. BACKGROUND

[0002] Photodynamic therapy has the characteristics of minimally invasive, high biocompatibility and high spatiotemporal selectivity, and has become one of the important medical technical means for treating malignant tumors.

[0003] Photodynamic therapy refers to that a drug is injected intravenously, the drug is selectively accumulated in tumor tissues, and after a specific time, a specific wavelength of light is used to irradiate the tumor tissues. At this time, the photosensitizer enriched in the tumor tissues will trigger a series of photophysical and chemical reactions under the excitation of light, and then produce active oxygen with cytotoxicity, so as to kill cancer cells and destroy tumor tissues. At present, a variety of photosensitizers have been applied to the research of cancer treatment. Rhodamine has been widely used in photodynamic therapy as a new photosensitizer for monitoring and diagnosis. In biological imaging, the distribution of photosensitizers in cells or tissues can be clearly observed, which helps doctors accurately determine the lesion site.

[0004] However, the absorption and emission wavelength of rhodamine is in the visible light region. Since the penetration of the wavelength in this region is weak, rhodamine is difficult to fully play a role in the treatment of deeper tissue sites. The existing technology usually expands the conjugated system in the molecule, promotes the delocalization and transmission of electrons in the molecule more effectively, causes the red shift of the absorption spectrum, enhances the absorption width of rhodamine to the near-infrared waveband, and makes rhodamine act on deeper tissue sites. However, the increase of the conjugated system may cause the aggregation phenomenon between molecules, so that the light absorption capacity of the aggregated photosensitizer molecules will be reduced, and then the yield of active oxygen substances such as singlet oxygen will be reduced, and the killing ability of cancer cells will also be weakened. SUMMARY

[0005] Therefore, the purpose of the present application is to provide a selenium-containing rhodamine derivative and application thereof, which can solve the problems of narrow absorption width of rhodamine to the near-infrared waveband and low yield of active oxygen substances.

[0006] To achieve the above technical purpose, the present application provides a selenium-containing rhodamine derivative, and the chemical structural formula 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-diethyl aniline substitution, 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 structure is one of the following,

[0020] .

[0021] The application provides a preparation method of rhodamine derivative A, comprising the following steps:

[0022] Step S1, under inert atmosphere, 10-ethyl-2-methoxy-10H-phenoselenoxazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-formyl) 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]phenoselenoxazine]-3-ketone;

[0023] Step S2, 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenoxazine]-3-ketone is dissolved in a solvent, phosphorus oxychloride is added, and heated to reflux to obtain a selenium-containing rhodamine derivative.

[0024] Further, in step S1, the reaction temperature is 50-80℃, the reaction time is 8-14 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenoxazine to 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℃, 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]phenoselenoxazine]-3-one to phosphorus oxychloride is 1:(2-3).

[0025] The application provides a preparation method of rhodamine derivative B, comprising the following steps:

[0026] In step S1, 10-ethyl-2-methoxy-10H-phenoselenoxazine and 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid are reacted to obtain 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenoxazine-14,1'-isobenzofuran]-3'-one under an inert atmosphere.

[0027] In step S2, 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenoxazine-14,1'-isobenzofuran]-3'-one is dissolved in a solvent, and phosphorus oxychloride is added and heated to reflux to obtain selenium-containing rhodamine derivative B.

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

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

[0030] The application provides an application of rhodamine derivative, which is used for treating malignant tumors.

[0031] Further, 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, the application innovatively proposes a rhodamine derivative containing selenium heavy atoms, which can extend the visible light absorption wavelength to above 600 NM, close to the near-infrared region of 700 NM. This characteristic 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 derivative can enhance the intersystem crossing rate in the molecule, so that the rhodamine derivative exhibits a high triplet state conversion rate under the excitation condition of light, generating a large number of active oxygen species to enhance the killing effect on cancer cells and cancer tissues. Compared with the prior art, the rhodamine derivative containing selenium heavy atoms of the application performs outstandingly in killing effect, and can achieve efficient removal of cancer cells and cancer tissues at a concentration of only nanomolar level, significantly improving the treatment efficiency and reducing the drug dosage.

[0033] The rhodamine derivative preparation method provided by the application has significant advantages. The synthesis route is simple, and only two main synthesis steps are needed to successfully prepare the target product. Compared with traditional complex synthesis techniques, the method not only has a simple synthesis process and is easy to implement, but also can obtain a high synthesis yield. This is not only beneficial to the rapid and low-cost preparation of the derivative at the laboratory scale, but also lays a solid foundation for subsequent large-scale industrial production, and has a wide application prospect and potential value in the field of drug research and production. BRIEF DESCRIPTION OF DRAWINGS

[0034] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the drawings needed to be used in the embodiments or the prior art description will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the application, and other drawings can be obtained by those skilled in the art without creative labor.

[0035] Figure 1 A chemical synthesis schematic diagram of the selenium-containing rhodamine derivative A provided for Embodiment 1 of the application is shown in the following figure:

[0036] Figure 2 A 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-ketone nuclear magnetic resonance hydrogen spectrum provided for Embodiment 1 of the application is shown in the following figure:

[0037] Figure 3 A nuclear magnetic resonance hydrogen spectrum of the selenium-containing rhodamine derivative A provided for Embodiment 1 of the application is shown in the following figure:

[0038] Figure 4 A chemical synthesis schematic diagram of the selenium-containing rhodamine derivative B provided for Embodiment 2 of the application is shown in the following figure:

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

[0040] Figure 6 NMR spectrum of selenium-containing rhodamine derivative B provided for Example 2 of the present application;

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

[0042] Figure 8 Active oxygen generation capacity of selenium-containing rhodamine derivatives provided for Examples 1-2 of the present application;

[0043] Figure 9 Growth inhibition of selenium-containing rhodamine derivatives on mouse breast cancer cells, human cervical cancer cells, and mouse melanoma cells provided for Examples 1-2 of the present application. DETAILED DESCRIPTION

[0044] The technical solutions of the embodiments of the present application will be described below in conjunction with the accompanying drawings, obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of the present application.

[0045] Among them, all raw materials of the present application have no special restrictions on their sources, and can be purchased on the market or prepared according to conventional methods well known to those skilled in the art.

[0046] The embodiments of the present application provide a selenium-containing rhodamine derivative, and the chemical structural formula is as follows:

[0047] .

[0048] It is particularly pointed out that the selenium-containing rhodamine derivative proposed in the present application is carefully designed according to the strategy of enhancing the killing ability of photosensitizers on cancer cells by selenium heavy atoms. After introducing selenium heavy atoms into its chemical formula, the derivative is endowed with unique optical and biological properties. From the optical properties, its visible light absorption wavelength is extended to the near-infrared band of 600-700 nm, which has a significant advantage in 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 state conversion rate of selenium-containing rhodamine derivatives, so that under light excitation, it can produce oxygen species with strong oxidative activity with high efficiency. From the biological properties, the small action radius and short lifetime of reactive oxygen species precisely limit their range of action. In the process of photodynamic therapy, only cancer cells that receive specific light irradiation are damaged, effectively avoiding unnecessary damage to surrounding normal cells, thereby minimizing the toxic side effects 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 allows them to easily cross the cell membrane and enter the cell interior to play a role, and the excellent biological tissue penetration provides a strong guarantee for their deep 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-diethyl aniline substitution, R2 is selected from ethyl, R3 is selected from a benzene ring, R4 is selected from an ester group, and R5 is selected from a hydrogen atom.

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

[0061] .

[0062] It should be noted that the SE-RHOD-A containing selenium rhodamine derivative exhibits the strongest visible light absorption wavelength of 610NM. Under light conditions, the selenium-containing rhodamine derivative of formula A can produce a significant killing effect on cells, especially for 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 application provides a preparation method of rhodamine derivative A, comprising the following steps:

[0064] Step S1, under an inert atmosphere, 10-ethyl-2-methoxy-10H-phenoselenoxazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-formyl) 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]phenoselenoxazine]-3-ketone.

[0065] Step S2, 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenoxazine]-3-ketone is dissolved in a solvent, phosphorus oxychloride is added, and heated to reflux to obtain a selenium-containing rhodamine derivative A.

[0066] In some embodiments, in step S1, the reaction temperature is 50-80℃, the reaction time is 8-14 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenoxazine to 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℃, 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]phenoselenoxazine]-3-ketone to phosphorus oxychloride is 1:(2-3).

[0067] The embodiment of the present application provides a preparation method of rhodamine derivative B, comprising the following steps:

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

[0069] Step S2, 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenoxazine-14,1'-isobenzofuran]-3'-one is dissolved in a solvent, phosphorus oxychloride is added, and heated to reflux to obtain selenium-containing rhodamine derivative B.

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

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

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

[0073] The applicant further provides the following reference specific embodiments to describe the application, and it should be noted that these embodiments are merely descriptive and do not limit the application in any way. Embodiment 1

[0074] Reference Figures 1-3 The embodiment provides a preparation method of a selenium-containing rhodamine derivative described in formula A, which comprises the following steps.

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

[0076] Argon gas was introduced into the chemical synthesis apparatus for protection. 10-Ethyl-2-methoxy-10H-phenselenazine (2g, 6.5 mmol), 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-formyl)benzoic acid (3g, 10 mmol), and 5 mL of methanesulfonic acid were added to the apparatus. The mixture was then heated to 70°C and refluxed for 12 hours. After reflux, heating was stopped and the mixture was cooled to room temperature. The final temperature was 40°C and maintained at -0.1 MPa. The solvent was removed by vacuum distillation at 5°C to obtain the intermediate. The intermediate was purified by silica gel column chromatography (eluting with PE:EA = 5:1) to give 1.5 G of blue solid, with a yield of 48%. The blue solid is 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromenone[2,3-B]phenoselenazine]-3-one, and its 1H NMR spectrum is shown below. Figure 2 As shown;

[0077] Step S2: Argon gas is introduced into the chemical synthesis apparatus for protection. 1g (1.8 mmol) of 13'-ethyl-1',2',3',4'-tetrahydro-3H,13'H-spiro[isobenzofuran-1,6'-pyrido[3',2':6,7]chromeno[2,3-B]phenoselenone]-3-one prepared in Step 1 and 10 mL of dichloromethane are placed in the chemical synthesis apparatus. Phosphorus oxychloride (600 mL, 6.4 mmol) is then added to the apparatus. The mixture is heated to 50°C and refluxed for 4 hours. After reflux is complete, heating is stopped and the mixture is cooled to room temperature. The solvent was removed by vacuum distillation at -0.1 MPa and 45°C to obtain an intermediate. 10 mL of dichloromethane, 890 mL of triethylamine (6.4 mmol), and 260 mL of methanol (6.4 mmol) were added to the intermediate, and the mixture was continuously stirred while the reaction was monitored in real-time by TLC. After the reaction was complete, a crude product was obtained. The crude product was then subjected to vacuum distillation at -0.1 MPa and 45°C to remove the solvent, followed by recrystallization with methanol as the solvent for purification, yielding 390 mg of a blue solid product A, with a yield of 45%. The 1H NMR spectrum is shown below. Figure 3 As shown. Example 2

[0078] See Figures 1-3 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[chromene[2,3-B]phenazine-14,1'-isobenzofuran]-3'-one:

[0080] The chemical synthesis device is protected by argon, 10-ethyl-2-methoxy-10H-phenoselenoxazine (2G, 6.5 mmol) and 2-(4-(diethylamino)-2-hydroxybenzoyl) benzoic acid (3.1G, 10 mmol), 5 ml of methanesulfonic acid are put into the synthesis device, then heated to 75 DEG C, and condensed to reflux for 10 hours. After the reflux is completed, the heating is stopped, and the temperature is cooled to room temperature. The solvent is removed by distillation under reduced pressure at a pressure of-0.1 MPa and a temperature of 45 DEG C to obtain an intermediate. The intermediate is purified by silica gel column chromatography (eluent: PE:EA = 3:1) to obtain 1.6G of blue solid with a yield of 43%. The blue solid is 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenoxazine-14,1'-isobenzofuran]-3'-one. The nuclear magnetic resonance hydrogen spectrum is shown in Figure 5

[0081] Step S2, the chemical synthesis device is protected by argon, 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-B]phenoxazine-14,1'-isobenzofuran]-3'-one (1G, 1.8 mmol) prepared in step 1 and 10 ml of dichloromethane are put into the synthesis device. Then, phosphorus oxychloride (600 ml, 6.4 mmol) is added to the synthesis device. After being heated to 50 DEG C, the mixture is condensed to reflux for 4 hours. After the reflux is completed, the heating is stopped, and the temperature is cooled to room temperature. The solvent is removed by distillation under reduced pressure at a pressure of-0.1 MPa and a temperature of 45 DEG C to obtain an intermediate. 10 ml of dichloromethane, triethylamine (890 ml, 6.4 mmol) and methanol (260 ml, 6.4 mmol) are added to the intermediate. The reaction process is monitored in real time by TLC. After the reaction is completed, the crude product is obtained. The solvent is removed by distillation under reduced pressure at a pressure of-0.1 MPa and a temperature of 45 DEG C. Then, the crude product is recrystallized in methanol to obtain 430 mg of blue solid product B with a yield of 45.5%. The nuclear magnetic resonance hydrogen spectrum is shown in Figure 6

[0082] The selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B prepared in examples 1 and 2 are subjected to performance tests to determine the effect of cell photodynamic therapy. The performance tests include spectral tests, active oxygen tests and cell photodynamic therapy tests.

[0083] Test 1: spectral test

[0084] The selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B are dissolved in different solvents, respectively, and subjected to ultraviolet-visible spectrophotometer tests. The results are shown in Figure 7 ​​The visible light absorption wavelength of the selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B involved in the present application approaches 700 NM, and the highest absorption intensity is shown at 590 NM and 610 NM, respectively. This indicates 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 the photodynamic therapy of deeper tissues.

[0085] Test 2: Active oxygen test

[0086] The selenium-containing rhodamine derivatives SE-RHOD-A or SE-RHOD-B are dissolved in a DMSO solvent, and then 1,3-diphenyl isobenzofuran (DPBF) is added to the solution to obtain a mixture. Then, the mixture is irradiated for a certain time length using red light with a wavelength range of 620-650 NM and an illumination intensity of 30 MW / CM², and the mixture is tested using a UV-visible spectrophotometer, focusing on recording the absorbance value of the mixture at a wavelength of 417 NM under different illumination times, and drawing a curve of the absorbance value of 1,3-diphenyl isobenzofuran versus time, as shown in Figure 8 .

[0087] When active oxygen is generated, 1,3-diphenyl isobenzofuran will undergo an irreversible oxidation reaction, at which time its absorption intensity at 417 NM will rapidly decrease. As can be clearly observed from the figure, the selenium-containing rhodamine derivatives SE-RHOD-A or SE-RHOD-B involved in the present application can rapidly generate active oxygen under light conditions, indicating that the selenium-containing rhodamine derivatives SE-RHOD-A and SE-RHOD-B have excellent photosensitivity. When they are applied in the field of photodynamic therapy, the rapidly generated active oxygen can effectively participate in the corresponding biochemical reactions, and play its therapeutic role.

[0088] Test 3: Cell photodynamic therapy test

[0089] Different cell lines are taken, and each is inoculated in a 96-well plate at a cell density of 5000 cells per well. The 96-well plate is placed in an incubator with a temperature of 37°C and a CO2 content of 5%, and incubated for 24 hours. The cell lines include human breast cancer cells (4T1 CELL), cervical cancer cells (HELA CELL), and melanoma cells (B16F10 CELL).

[0090] The selenium-containing 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, and different concentrations of the mixed solutions were added to each well and incubated in a cell culture incubator for 2 hours. The mixed solution in the well plate was removed with a pipette displacement gun, and fresh mixed solution was added, and the well plate was placed in red light with a wavelength range of 620-650 NM and an illumination intensity of 30 MW / CM² for 10 minutes. After irradiation, the well plate was placed in the cell culture incubator for 22 hours, then the mixed solution in the well plate was removed with a pipette displacement gun, and 150 UL of MTT medium with a concentration of 5 MG / ML was added to the well plate, and the well plate was moved to the cell culture incubator for incubation for 4 hours. The MTT medium in the plate well was removed, and then 100 UL of DMSO was added to dissolve the oxidation product of MTT. The absorbance of each well solution was measured at 490 NM wavelength using an enzyme marker, and the cell survival rate was calculated based on the linear relationship between absorbance and cell number, as shown in Tables 1-3. According to the cell survival rate data of SE-RHOD-A and SE-RHOD-B in different cell lines, the IC 50 values of SE-RHOD-A or SE-RHOD-B were calculated by fitting, and the fitting results are shown in Figure 9 .

[0091] Table 1

[0092]

[0093] According to the data in Table 1 and Figure 9 , 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 can be apoptotic.

[0094] Table 2

[0095]

[0096] According to the data in Table 2 and Figure 9The data show that when the SE-RNOD-A concentration was 0.12 μM, the survival rate of HELA cells was 19.69%; when the SE-RNOD-A concentration was 1.1 μM, the survival rate was as low as 12.67%. When the SE-RNOD-B concentration was 0.12 μM, the survival rate of HELA cells was 11.27%; when the SE-RNOD-B concentration was 3.3 μM, the survival rate was as low as 6.36%. When the SE-RNOD-A and SE-RNOD-B dosages were 31 nmM and 13 nmM, respectively, apoptosis of 50% of HELA cells could be achieved.

[0097] Table 3

[0098]

[0099] According to Table 3 and Figure 9 The data show that when the SE-RNOD-A concentration was 0.22 μM, the survival rate of B16F10 cells was 18.68%; when the SE-RNOD-A concentration was 2.0 μM, the survival rate was 13.75%. When the SE-RNOD-B concentration was 0.22 μM, the survival rate of B16F10 cells was 14.69%; when the SE-RNOD-B concentration was 2.0 μM, the survival rate was 11.59%. When the SE-RNOD-A and SE-RNOD-B dosages were 55 nmM and 52 nmM, respectively, apoptosis of 50% of B16F10 cells could be achieved.

[0100] In summary, the selenium-containing rhodamine derivative provided in this application exhibits significant killing efficacy against various cancer cells under light irradiation. Within the scope of photodynamic therapy for malignant tumors, it demonstrates considerable potential and application value, opening up a promising new photosensitizer pathway for tumor treatment.

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

Claims

1. A selenium-containing rhodamine derivative, characterized by, The chemical structural formula is as follows: ； The R1 substituent is one of the following groups: ； The R2 substituent is one of the following groups, ； The R3 substituent is one of the following groups, ； The R4 substituent is one of the following groups, ； The R5 substituent is one of the following groups, 。 2. The selenium-containing rhodamine derivative according to claim 1, characterized in that, The R1 substituent is selected from 1,2,3,4-tetrahydroquinoline or N,N-diethyl aniline, the R2 substituent is selected from ethyl, the R3 substituent is selected from a benzene ring, the R4 substituent is selected from a methyl ester group, and the R5 substituent is selected from a hydrogen atom.

3. The selenium-containing rhodamine derivative according to claim 2, characterized in that, The chemical structural formula is one of the following, 。 4. A method for preparing rhodamine derivative Se-Rhod-A according to claim 3, characterized by, The method comprises the following steps: Step S1: reacting 10-ethyl-2-methoxy-10H-phenoselenoxazine and 2-(7-hydroxy-1,2,3,4-tetrahydroquinoline-6-formyl)benzoic acid under an inert atmosphere 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]phenoselenoxazine]-3-ketone; 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]phenoselenoxazine]-3-ketone in methanol, adding phosphorus oxychloride, and heating to reflux to obtain the selenium-containing rhodamine derivative Se-Rhod-A; In the preparation method of the rhodamine derivative Se-Rhod-A, the conversion process of the chemical structural formula is as follows: 。 5. The preparation method of the rhodamine derivative Se-Rhod-A according to claim 4, characterized in that, In the step S1, the reaction temperature is 50-80 DEG C, the reaction time is 8-14 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenoxazine 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 DEG 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]phenoselenoxazine]-3-ketone to phosphorus oxychloride is 1:(2-3).

6. A method for preparing rhodamine derivative Se-Rhod-B according to claim 3, characterized by, The method comprises the following steps: Step S1: reacting 10-ethyl-2-methoxy-10H-phenoselenoxazine and 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid under an inert atmosphere to obtain 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-b]phenoxazine-14,1'-isobenzofuran]-3'-ketone; Step S2: dissolving 3-(diethylamino)-7-ethyl-3'H,7H-spiro[chromeno[2,3-b]phenoxazine-14,1'-isobenzofuran]-3'-ketone in methanol, adding phosphorus oxychloride, and heating to reflux to obtain the selenium-containing rhodamine derivative Se-Rhod-B; The conversion process of the chemical structural formula in the preparation method of the rhodamine derivative Se-Rhod-B is as follows: 。 7. The preparation method of the rhodamine derivative Se-Rhod-B according to claim 6, characterized in that, In the step S1, the reaction temperature is 60-80 DEG C, the reaction time is 10-12 hours, and the molar ratio of 10-ethyl-2-methoxy-10H-phenoselenoxazine and 2-(4-(diethylamino)-2-hydroxybenzoyl)benzoic acid is 1:(1-3); In the step S2, the reaction temperature is 0-50 DEG 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]phenoselenoxazine]-3-one and phosphorus oxychloride is 1:(2-3).

8. Use of a rhodamine derivative according to any one of claims 1 to 3, characterized in that, A drug for treating malignant tumors.

9. The use of a rhodamine derivative according to claim 8, characterized in that, The malignant tumors are 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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