Application of quetiapine fumarate in preparing medicine for treating retinoblastoma

Abstract

The present invention discloses the use of quetiapine fumarate in the preparation of a drug for treating retinoblastoma. The present invention is the first to discover a new role of quetiapine fumarate in the treatment of retinoblastoma (Rb). Quetiapine fumarate can significantly inhibit the growth of Rb, promote the apoptosis of tumor cells, and effectively treat Rb. The present invention utilizes an Rb organoid model obtained by in vitro induced differentiation of pluripotent stem cells to discover a new role of quetiapine fumarate in the treatment of Rb, providing a new approach to the treatment of Rb. The present invention aims to provide a new drug for the treatment of Rb, improve the effectiveness of tumor treatment, expand the application range of quetiapine fumarate, and provide a reference for clinical medication.

Description

Technical Field

[0001] The present invention relates to the field of medicine, in particular to application of quetiapine fumarate in preparing medicine for treating retinoblastoma. Background Art

[0002] Retinoblastoma (Rb) is the most common intraocular malignancy in children worldwide and the third most common childhood cancer. Rb is caused by pathogenic mutations in both alleles of the RB1 gene during retinal cell development. It is aggressive and poses a serious threat to children's vision and quality of life. Due to medical, financial, and educational limitations, many children seek medical attention in the advanced stages of the disease, resulting in extremely high rates of blindness and disability.

[0003] The most common treatments for Rb include systemic and local chemotherapy, laser photocoagulation, cryotherapy, surgical resection, and brachytherapy. For patients with advanced Rb, most developing countries use chemotherapy-based combination therapies. However, chemotherapy has toxic side effects, and the development of chemotherapy resistance leads to treatment failure in some children. Radiotherapy increases the risk of second primary tumors. Local treatments are prone to tumor recurrence. Consequently, this disease places a significant burden on both families and society. There is an urgent need to identify new treatments to address this issue.

[0004] Recent studies have found that hESCs with RB1 gene mutations or knockouts can be differentiated into Rb organoids in vitro. The construction of this organoid model provides a reliable model for the study of Rb pathogenesis, drug development and treatment.

[0005] Quetiapine fumarate has affinity for multiple neurotransmitter receptors (including serotonin, dopamine, histamine, and adrenergic receptors) and is an atypical antipsychotic used to treat schizophrenia, bipolar I mania, bipolar II depression, and bipolar I depression. Currently, there are no reports of its use in the treatment of Rb. Summary of the Invention

[0006] In view of this, the present invention proposes the use of quetiapine fumarate in the preparation of a medicament for treating retinoblastoma (Rb).

[0007] The structure of quetiapine fumarate is shown in Formula I:

[0008]

[0009] The retinoblastoma is: a cell line with homozygous mutation of RB1 gene (hESCs-RB1 mut / mut) Retinoblastoma organoids formed after 75 to 90 days of differentiation culture.

[0010] The final concentration of quetiapine fumarate in the drug for treating retinoblastoma is greater than or equal to 10 μM.

[0011] The final concentration of quetiapine fumarate in the drug for treating retinoblastoma is 10 μM to 40 μM.

[0012] The drug for treating retinoblastoma is administered for 3 to 7 days.

[0013] The quetiapine fumarate inhibits the proliferation of retinoblastoma cells by reducing the expression of Ki67, a proliferative cell marker in the tumor area; and / or the quetiapine fumarate promotes the apoptosis of retinoblastoma cells by increasing the expression of Cleaved caspase 3, a apoptotic cell marker.

[0014] The present invention also provides the use of quetiapine fumarate in preparing a medicine for inhibiting the growth of tumor tissue in an Rb cell line Y79 xenograft tumor animal model.

[0015] The final concentration of quetiapine fumarate in the drug is 20 mg / kg.

[0016] The drug is administered by intraperitoneal injection.

[0017] Quetiapine fumarate is a non-classical antipsychotic drug, mainly used to treat schizophrenia. The applicant found in the study that quetiapine fumarate has a very good therapeutic effect on Rb: (1) Figure 6 As can be seen in the figure, after the Rb organoids were treated with quetiapine fumarate, the tumor site no longer proliferated and even gradually became smaller as the culture time prolonged. (2) Figures 7-10 It can be seen that with the increase of drug concentration and the extension of time, the proliferating cells in the tumor area gradually decreased and the apoptotic cells gradually increased, proving that quetiapine fumarate can effectively inhibit the proliferation of Rb and promote tumor apoptosis.

[0018] This invention, for the first time, discovers a novel role for quetiapine fumarate in the treatment of Rb. Quetiapine fumarate can significantly inhibit Rb growth and promote tumor cell apoptosis, effectively treating Rb. This invention utilizes an Rb organoid model derived from in vitro induced differentiation of pluripotent stem cells to discover a novel role for quetiapine fumarate in the treatment of Rb, providing new insights into the treatment of Rb. This invention aims to provide new drugs for the treatment of Rb, improve the effectiveness of tumor treatment, expand the application range of quetiapine fumarate, and provide a reference for clinical medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] For purposes of illustration and not limitation, the present invention will now be described with reference to preferred embodiments thereof, particularly with reference to the accompanying drawings, in which:

[0020] Figure 1 It is a flow chart for testing the effectiveness and safety verification of tumor drugs.

[0021] Figure 2 This is the result of validating the effectiveness of the drug quetiapine fumarate using the Rb cell line Y-79; Figure 2 Middle A is the CCK8 standard curve of the Rb cell line (Y79); Figure 2 Middle B is the CCK8 cell viability test after quetiapine fumarate treatment for 24 hours; the statistical results were analyzed by one-way ANOVA, Mean±SD, ***P<0.001, n=3.

[0022] Figure 3 The results of the safety validation of the drug quetiapine fumarate were conducted using the retinal pigment epithelial cell line (ARPE19); Figure 3 Middle A is the CCK8 standard curve of ARPE19 cell line;

[0023] Figure 3 Middle B is the CCK8 cell viability test after quetiapine fumarate treatment for 24 hours; the statistical results were analyzed by one-way ANOVA, Mean±SD, ns p>0.05, n=3.

[0024] Figure 4 The safety test results of the drug quetiapine fumarate were conducted using retinal progenitor cells (RPCs) differentiated from human embryonic stem cells. Figure 4 Middle A is the CCK8 standard curve of RPCs; Figure 4 Middle B is the CCK8 cell viability test after quetiapine fumarate treatment for 24 hours; the statistical results were analyzed by one-way ANOVA, Mean±SD, ns p>0.05, n=3.

[0025] Figure 5 The safety test results of the drug quetiapine fumarate were conducted using primary human neonatal fibroblasts (hPFs); Figure 5 Middle A is the CCK8 standard curve of hPFs; Figure 5 Middle B is the CCK8 cell viability test after quetiapine fumarate treatment for 24 hours; the statistical results were analyzed by one-way ANOVA, Mean±SD, ns p>0.05, n=3.

[0026] Figure 6 Quetiapine fumarate is used to treat Rb organoids. Rb organoids containing tumors on the 75th day of differentiation were treated with different drug concentrations, and the morphological changes of Rb organoids were observed under a microscope after 3 or 7 days of drug treatment.

[0027] Figure 7 This is the immunofluorescence staining result of proliferation / apoptosis-related markers of Rb organoids after quetiapine fumarate treatment for 3 days; Ki67, proliferation marker; Cleaved Caspase 3, apoptosis marker; Merge, multi-channel overlay; Scale bar, 50 μm.

[0028] Figure 8 Figure 3 shows immunofluorescence staining of proliferation and apoptosis markers in Rb organoids treated with quetiapine fumarate for 3 days; Ki67, a proliferation marker; and Cleaved Caspase 3, an apoptosis marker. Statistical results were analyzed using one-way ANOVA (mean ± SD). #p>0.05, *P<0.05, **P<0.01, ***P<0.001; n = 3.

[0029] Figure 9 This is the immunofluorescence staining result of proliferation / apoptosis-related markers of Rb organoids after 7 days of quetiapine fumarate treatment; Ki67, proliferation marker; Cleaved Caspase 3, apoptosis marker; Merge, multi-channel overlay; Scale bar, 50 μm.

[0030] Figure 10 Figure 2 shows immunofluorescence staining of proliferation and apoptosis markers in Rb organoids treated with quetiapine fumarate for 7 days. Ki67 is a proliferation marker; Cleaved Caspase 3 is an apoptosis marker. Statistical results were analyzed using one-way ANOVA (mean ± SD). #p>0.05, *P<0.05, **P<0.01, ***P<0.001; n = 3).

[0031] Figure 11 Intraperitoneal injection of quetiapine fumarate was used to treat subcutaneous transplanted tumors in immunodeficient mice. The tumors in the drug treatment group were smaller than those in the control group. DETAILED DESCRIPTION

[0032] The embodiments of the present invention will be described in detail below with reference to the examples. However, it should be understood by those skilled in the art that the following examples are merely illustrative of the present invention and should not be construed as limiting the scope of the present invention. The reagents or instruments used in the examples, unless their sources are indicated, are all commercially available conventional products.

[0033] Example 1: Use of Quetiapine Fumarate in the Preparation of a Drug for Treating Retinoblastoma Figure 1 This is a flow chart for validation of the effectiveness and safety of tumor drugs.

[0034] 1. Construction of a retinoblastoma (Rb) disease model

[0035] A human embryonic stem cell line harboring an RB1 gene mutation was constructed, and Rb organoids were obtained using in vitro retinal organoid differentiation technology. The construction method is described in the following literature:

[0036] 1.Li YP, Wang YT, Wang W, et al. Second hit impels oncogenesis ofretinoblastoma in patient-induced pluripotent stem cell-derived retinalorganoids: direct evidence for Knudson's theory.Furmanski P,ed.PNASNexus.2022;1(4):pgac162.doi:10.1093 / pnasnexus / pgac162.

[0037] 2. Liu H, Zhang Y, Zhang YY, et al. Human embryonic stem cell-derivedorganoid retinoblastoma reveals a cancerous origin. Proc Natl Acad SciUSA.2020;117(52):33628-33638.doi:10.1073 / pnas.2011780117.

[0038] 3.Pan D,

[0039] (1) Establishment of RB1 gene mutation stem cell line. The construction method is described in the following literature:

[0040] Liu H, Zhang Y, Zhang YY, et al. Human embryonic stem cell-derivedorganoid retinoblastoma reveals a cancerous origin. Proc Natl Acad SciUSA.2020;117(52):33628-33638.doi:10.1073 / pnas.2011780117.

[0041] Construction of RB1 targeting vector: Using CRISPR / Cas9 gene editing technology, c.958C>T; p.R320X point mutations were made in the RB1 gene of human embryonic stem cell line (hESCs) (WiCell) to construct a cell line with homozygous RB1 mutation (hESCs-RB1 mut / mut sgRNA was first designed using the CRISPR design tool http: / / crispr.mit.edu / edu / . Subsequently, the activity of sgRNAs was evaluated by the universal CRISPR RNA activity assay (Biocygen). Active sgRNAs were cloned and used to construct CRISPR / Cas9 plasmids (CN110241138 A).

[0042] Establishment of RB1 gene mutation cell line: hESCs were dissociated into single cells using TrypLE Select digestion solution (Life Technology). 1×10 6 The cells were suspended in electroporation solution (LONZA) containing plasmids and subjected to nuclear transfection using an electroporator. After nuclear transfection, the cells were cultured in an incubator for 2 to 3 days. They were treated with 2 μg / mL puromycin (Shanghai Qianchen, ISY1130), and clones resistant to puromycin were picked and amplified (CN110241138 A) to obtain a cell line with homozygous RB1 mutation (hESCs-RB1 mut / mut ).

[0043] (2) Construction of Rb organoids

[0044] The RB1 homozygous mutant cell line obtained above (hESCs-RB1 mut / mut) were digested into single cells using TrypLE Select containing 0.05 mg / mL DNaseI (Roche, 11284932001) and 20 μM Y-27632 (Selleck, S1049); the cells were cultured using type I differentiation medium (containing 80% GMEM (Gibco, 11710), 20% KSR (ThermoFisherScientific, 10828028), 1% PS (gibco, 15140-122), 1 mM NEAA (sigma, M7145), 1 mM pyruvate (sigma, S8636), 100 μM β-mercaptoethanol (sigma, M3148), 3 μM Wnt Antagonist I (Merck Millipore, 681669)) was used to adjust the cell concentration to 120,000 cells / mL and seeded into a low-adhesion 96-well V-plate at 100 μL / well; on the second day, Matrigel (BD.corning, 356231) was added to the 96-well V-plate to a volume ratio of 1% (v / v); on the sixth day, half of the type I differentiation medium was replaced with fresh one; on the 12th day, the cell clusters were picked out and placed in a 10 cm non-adhesive culture dish, and replaced with type II differentiation medium (containing 90% GMEM, 10% FBS (BI, 04-001-1A), 1% PS, 1 mM NEAA, 1 mM pyruvate, 100 μM β-mercaptoethanol, 100 nM SAG (Enzo Life Sciences, ALX-270-426-M001), and 1% Matrigel). On day 18, the organoids were divided into four sections using an ophthalmic scalpel under a microscope, and the culture medium was replaced with NR medium (80% DMEM / F12 (Gibco, 10565-018), 10% FBS, 1% N-2, 0.5 mM retinoic acid (Sigma, R7882), 0.1 mM taurine (Sigma, T8691), 1% PS). Tumor primaries began to appear in the organoids on day 45 of differentiation culture, and the tumors gradually increased in size with increasing differentiation time. By day 90, Rb organoids had formed. Organoid cultures were maintained in a 37°C, 5% CO2 cell culture incubator.

[0045] 2. Verification of drug effectiveness using Rb cell lines

[0046] The efficacy of quetiapine fumarate was tested on the Rb cell line (Y79) (ATCC, Cat#HTB-18), and cell viability and cytotoxicity were determined using CCK8.

[0047] Effectiveness drug determination: The CCK8 method was used to detect the cytotoxicity of quetiapine fumarate on Y79 cells.

[0048] The specific steps are as follows:

[0049] 1. Y79 cell suspension was seeded into 96-well plates at a cell count of 7500 cells / 100 μL per well. A blank control group was added with only Y79 medium (containing 79% RPMI-1640 (Sigma, R0883-500ML), 20% FBS, and 1% PS) and cultured in a 37°C, 5% CO2 cell culture incubator for 24 hours.

[0050] 2. Add quetiapine fumarate to the Y79 culture medium to a final concentration of 20 μM. For the negative control, add dimethyl sulfoxide (DMSO) and culture in a 37°C, 5% CO2 cell culture incubator for 24 hours.

[0051] 3. Add 10 μL of CCK8 to each well of a 96-well plate, incubate at 37°C, 5% CO2 for 4 hours, and measure the absorbance at 450 nm.

[0052] Cytotoxicity was defined as relative survival, with the survival rate of cells in the culture medium without quetiapine fumarate being 100% ( Figure 2 ). Figure 2 Middle A is the CCK8 standard curve of the Rb cell line (Y79);

[0053] Figure 2 Figure B shows the CCK8 cell viability assay results after 24 hours of quetiapine fumarate treatment; statistical results were analyzed using one-way ANOVA, Mean ± SD, ***P < 0.001, n = 3. The experimental results show that after 24 hours of quetiapine fumarate treatment, the CCK8 assay for Y79 cell viability was much lower than that of the dimethyl sulfoxide group, suggesting that quetiapine fumarate can inhibit the growth of Rb cells ( Figure 2 ).

[0054] 3. Verifying Drug Safety

[0055] The safety of quetiapine fumarate was verified for cells of different sources (commercial retinal pigment epithelial cell line (ARPE19) (ATCC, Cat#CRL-2302), primary human neonatal fibroblasts (hPFs) (ATCC, PCS-201-010), and retinal progenitor cells (RPCs) derived from hESCs). The construction method of retinal progenitor cells (RPCs) derived from hESCs was described in the following literature: Lamba DA, Karl MO, Ware CB, et al. Efficient generation of retinal progenitor cells from human embryonic stem cells [J]. Proceedings of the National Academy of Sciences of the United States of America, 2006, 103(34): 12769-12774., and cell activity and cytotoxicity were determined using CCK8. ARPE19 and hPFs were cultured in a standard cell culture medium (89% DMEM basic (Gibco, C11995500bt) + 10% FBS + 1% PS). RPCs were cultured in a medium containing DMEM / F12, B-27 (Gibco, 0080085-SA), N-2, 10 ng / ml mouse noggin (R&D systems), 10 ng / ml human recombinant Dkk-1 (R&D systems), 10 ng / ml human recombinant IGF-1 (R&D systems), and 5 ng / ml human recombinant bFGF (R&D Systems).

[0056] Safety drug selection: CCK8 method was used to detect the cytotoxicity of quetiapine fumarate to various cells.

[0057] The specific steps are as follows:

[0058] 1. Inoculate the cell suspension into a 96-well plate at a cell count of 4500 cells / 100 μL per well. Add only culture medium as the blank group and incubate at 37°C, 5% CO2 in a cell culture incubator for 24 hours.

[0059] 2. Add quetiapine fumarate to the culture medium. Add dimethyl sulfoxide to the negative control. Incubate at 37°C, 5% CO2 in a cell culture incubator for 24 hours.

[0060] 3. Add 10 μL of CCK8 to each well of a 96-well plate, incubate at 37°C, 5% CO2 for 4 hours, and measure the absorbance at 450 nm.

[0061] Cytotoxicity was defined as relative survival, with the survival rate of cells in the culture medium without quetiapine fumarate being 100% ( Figure 3-5 ). Figure 3 The results of the safety validation of the drug quetiapine fumarate were conducted using the retinal pigment epithelial cell line (ARPE19); Figure 3 Middle A is the CCK8 standard curve of ARPE19 cell line; Figure 3 Middle B is the result of CCK8 cell viability detection after quetiapine fumarate treatment for 24 hours; the statistical results were analyzed by one-way ANOVA, Mean±SD, ns p>0.05, n=3. Figure 4 The safety test results of the drug quetiapine fumarate were conducted using retinal progenitor cells (RPCs) differentiated from human embryonic stem cells. Figure 4 Middle A is the CCK8 standard curve of RPCs; Figure 4 Middle B is the result of CCK8 cell viability detection after quetiapine fumarate treatment for 24 hours; the statistical results were analyzed by one-way ANOVA, Mean±SD, ns p>0.05, n=3. Figure 5 The safety test results of the drug quetiapine fumarate were conducted using primary human neonatal fibroblasts (hPFs); Figure 5 Middle A is the CCK8 standard curve of hPFs; Figure 5 Middle B is the CCK8 cell viability test after quetiapine fumarate treatment for 24 hours; the statistical results were analyzed by one-way ANOVA, Mean±SD, ns p>0.05, n=3.

[0062] The experimental results show that after 24 hours of treatment with quetiapine fumarate, the cell viability was detected by CCK8. There was no statistical difference in cell viability between the group treated with dimethyl sulfoxide, indicating that the addition of quetiapine fumarate did not affect the retinal pigment epithelial cell line (ARPE19) ( Figure 3 ), hESCs-derived retinal progenitor cells (RPCs) ( Figure 4 ) and the growth of primary human neonatal fibroblasts (hPFs) ( Figure 5 ).

[0063] IV. Drug Intervention Therapy in Rb Organoid Models

[0064] The therapeutic study of quetiapine fumarate was conducted on Rb organoids derived from human pluripotent stem cells in vitro. The specific method is as follows:

[0065] ① Take Rb organoids with a differentiation time of 75 to 90 days and divide them into ten groups;

[0066] ② Five groups received short-term medication for 3 days. Drugs were added to NR culture medium and diluted to specific concentrations, including 0 μM (dimethyl sulfoxide group (Sigma, D2650)), 10 μM, 20 μM, 30 μM, and 40 μM.

[0067] ③ The other five groups received long-term medication for 7 days. Drugs were added to NR culture medium and diluted to specific concentrations. The drug concentrations were 0 μM (DMSO group), 10 μM, 20 μM, 30 μM, and 40 μM, respectively.

[0068] 5. Evaluation of tumor suppression effect

[0069] ① Record tumor growth every day using bright field microscopy. The experimental results are as follows: Figure 6 As shown, after the Rb organoids were treated with quetiapine fumarate, the Rb part no longer grew, whether it was 3 days or 7 days ( Figure 6 ).

[0070] ② Collect Rb organoids, fix them, embed them for frozen sections, and use immunofluorescence staining to detect the expression of proliferation / apoptosis markers in the tumor site and normal site in the tumor organoids.

[0071] The specific steps of immunofluorescence staining are as follows:

[0072] 1. Take out the frozen sections from -80℃ and let them stand at room temperature for 30 minutes. During this time, prepare the blocking solution (0.5%

[0073] Triton X-100 and 4% BSA 1:1) and incubation solution (0.5% Triton X-100 and 1% BSA 1:1) were prepared.

[0074] 2. After rewarming, wash the frozen sections with PBS buffer three times, 10 minutes each time. After washing, use an immunohistochemistry pen to draw a circle around the sample, add 50 μL of blocking solution into the circle, and let it stand at room temperature for 1 hour.

[0075] 3. After blocking, wash with PBS three times, 10 minutes each time.

[0076] 4. Prepare the primary antibody using the incubation solution according to the dilution ratio. Add 50 μL of primary antibody to each circle and incubate in a 4°C refrigerator overnight (8-12 hours).

[0077] 5. The next day, place the sections at room temperature for 30 minutes, recover the primary antibody, and then wash with PBS three times for 10 minutes each time.

[0078] 6. During this period, select the secondary antibody according to the species properties of the primary antibody used in the experiment, and prepare the secondary antibody in incubation solution at a dilution ratio of 1:400.

[0079] 7. Add 50 μL of secondary antibody to each circle, protect from light, and incubate at room temperature for 1 hour.

[0080] 8. Recover the secondary antibody, store in the dark, and wash with PBS three times, 10 minutes each time.

[0081] 9. During this period, prepare 10X DAPI with DPBS buffer, add 50 μL to each circle, protect from light, incubate at room temperature for 10 minutes, and wash with PBS three times, each time for 10 minutes.

[0082] 10. After the slices are dried, add an appropriate amount of anti-fluorescence quenching agent, cover with a coverslip, and store the slices in the dark at 4°C or observe them under a confocal microscope as soon as possible.

[0083] The experimental results are as follows: after the Rb organoids were treated with quetiapine fumarate, the number of Ki67-positive cells, a proliferative cell marker, in the tumor area gradually decreased with the increase of drug concentration, while the number of Cleaved caspase 3-positive cells, apoptotic cell marker, gradually increased with the increase of drug concentration ( Figure 7-10 ), especially long-term treatment (7 days) ( Figure 9-10 After 3 days of treatment, there was no significant difference between the drug concentrations of 30 μM and 40 μM in the bar graph ( Figure 8 ).

[0084] 6. Animal Experimentation

[0085] ① Rb cell line Y79 (1*10 5 cells / μL) were injected subcutaneously into immunodeficient mice (Shanghai Model Organisms, NM-KO-00158) to establish a xenograft tumor animal model;

[0086] ② One week later, the experimental group (4 mice) were intraperitoneally injected with quetiapine fumarate (20 mg / kg, prepared in corn oil (Selleck, S6701)); the control group (4 mice) were intraperitoneally injected with an equal volume of corn oil, and then subcutaneously injected every other day. The preparation method is as follows:

[0087] (1) Prepare the stock solution: dissolve 0.5 mg of quetiapine fumarate in 10 μL of dimethyl sulfoxide (DMSO) to a stock solution concentration of 50 mg / mL;

[0088] (2) Prepare the working solution: Dissolve 10 μL of the stock solution in 190 μL of corn oil and mix thoroughly. The working solution concentration is 2.5 mg / mL.

[0089] ③Record the weight of mice and measure the size of tumors every day;

[0090] ④ Four weeks later, the mice were killed, the tumor tissues were taken, and the tumor volume was weighed.

[0091] The experimental results are as follows Figure 11 As shown, the tumor tissue of the intraperitoneal injection of quetiapine fumarate was significantly smaller than that of the control group (intraperitoneal injection of corn oil).

[0092] The above specific embodiments do not limit the scope of protection of the present invention. Those skilled in the art will appreciate that various modifications, combinations, sub-combinations, and substitutions may occur depending on design requirements and other factors. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention are intended to be included within the scope of protection of the present invention.

Claims

1. Use of quetiapine fumarate in the preparation of a medicament for treating retinoblastoma.

2. The use according to claim 1, characterized in that: The structure of quetiapine fumarate is shown in Formula I:

3. The use according to claim 1, characterized in that: The retinoblastoma is a retinoblastoma organoid formed by differentiating and culturing a cell line with homozygous mutation of the RB1 gene for 75 to 90 days.

4. The use according to any one of claims 1 to 3, characterized in that: The final concentration of quetiapine fumarate in the drug for treating retinoblastoma is greater than or equal to 10 μM.

5. The use according to claim 4, characterized in that: The final concentration of quetiapine fumarate in the drug for treating retinoblastoma is 10 μM to 40 μM.

6. The use according to claim 5, characterized in that: The drug for treating retinoblastoma is administered for 3 to 7 days.

7. The use according to any one of claims 1 to 3, characterized in that: The quetiapine fumarate inhibits the proliferation of retinoblastoma cells by reducing the expression of Ki67, a proliferative cell marker in the tumor area; and / or The quetiapine fumarate promotes the apoptosis of retinoblastoma cells by increasing the expression of the apoptotic cell marker Cleaved caspase 3.

8. Use of quetiapine fumarate in the preparation of a drug for inhibiting tumor tissue growth in an Rb cell line Y79 xenograft tumor animal model.

9. The use according to claim 8, characterized in that: The final concentration of quetiapine fumarate in the drug is 20 mg / kg.

10. The use according to claim 8 or 9, characterized in that: The drug is administered by intraperitoneal injection.

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