Application of novel SRSF1 splicing factor small molecule inhibitor okanin and pharmaceutical composition thereof

Abstract

The application discloses application of a novel SRSF1 splicing factor small molecule inhibitor Okanin and a medicine composition thereof, wherein the medicine composition comprises an active ingredient, and the active ingredient comprises the SRSF1 splicing factor small molecule inhibitor Okanin and an immune checkpoint inhibitor alpha PD-1; the direct action target of Okanin is an RNA binding core region of SRSF1 protein, the downstream glycolysis function of SRSF1 is prevented from being exerted, and thus tumor cell proliferation is inhibited. Meanwhile, Okanin can act on CD8+ T cells, the effector function of the CD8+ T cells is enhanced, and thus the activity of the CD8+ T cells in killing tumor cells is improved.

Description

Technical Field

[0001] This invention relates to the fields of biomedicine and molecular biology, and in particular to the application of a novel small molecule inhibitor of SRSF1 cleavage factor, Okanin, and its drug combination. Background Technology

[0002] Although immune checkpoint blockade (αPD1) has revolutionized cancer treatment, only a small percentage of cancer patients exhibit durable clinical responses. Therefore, there is significant interest in identifying the mechanisms of pre-existing and acquired immune resistance and developing novel treatments to prevent relapse. Researchers believe that challenges such as low response rates and acquired resistance persist, primarily because activation of tumor-inherent pathways promotes immune escape or the depletion of cytotoxic T cells. While many anticancer drugs that can induce substantial tumor shrinkage have been developed by inhibiting key signaling pathways required for cell proliferation, relapse remains a major challenge due to the growth of drug-resistant tumor cells. The immune system may target residual disease, but many tumor cell-targeting drugs also impair the survival / function of immune cells. Therefore, it is crucial to develop tumor cell-targeting drugs that enhance rather than impair immune function. Two rate-limiting steps in the cancer immune cycle have been reported: T cell transport to the tumor and overcoming T cell exhaustion in the tumor microenvironment (TME). Recent studies have highlighted the role of tumor and T cell metabolism in cancer development and immunotherapy resistance. Metabolic restrictions imposed in the TME, such as glucose deprivation and hypoxia, dramatically alter cellular signaling in tumor-infiltrating lymphocytes, leading to impaired antitumor immune responses. Furthermore, exhausted T cells exhibit suppressed glycolysis and / or mitochondrial respiration, and this poor metabolic adaptation may exacerbate T cell exhaustion. Therefore, metabolic interventions to enhance the effector function of exhausted T cells urgently need to be addressed.

[0003] This study focuses on serine-arginine-rich splicing factor 1 (SRSF1), a serine / arginine-rich (SR) splicing factor (SF) involved in alternative splicing (AS). Some studies have shown that SRSF1-induced aberrant AS events include promoting systemic lupus erythematosus (SLE) tumorigenesis and controlling pathological changes in T cell phenotype. Furthermore, targeting SRSF1 is considered a potential therapeutic target for solid tumors. Recently, the inventors discovered a network regulatory relationship between SFs and metabolic-related pathways and immune responses in HCC through dysregulated SF-induced precursor mRNA transcripts. However, whether SRSF1 expression within tumors and T cells can control the immune environment of the tumor microenvironment (TME), reprogram the T cell metabolic profile, and restore the function of exhausted T cells remains to be explored. Summary of the Invention

[0004] This invention provides the application of Okanin, a novel small molecule inhibitor of SRSF1 cleavage factor, and its drug combination.

[0005] The application of the novel SRSF1 cleavage factor small molecule inhibitor Okanin is characterized by:

[0006] This includes the application of reagents for detecting the expression of the cleavage factor SRSF1 in products for prognostic assessment of liver cancer;

[0007] Products that downregulate tumor cell glycolysis and enhance the killing activity of CD8+ T cells;

[0008] Immune checkpoint inhibitors that enhance immunity or products that prepare immune checkpoint inhibitors;

[0009] Reversing tumor immune escape or preparing products that reverse tumor immune escape;

[0010] Products that inhibit tumor resistance to immune checkpoint inhibitors.

[0011] As a preferred technical solution, the tumor includes hepatocellular carcinoma and melanoma.

[0012] As a preferred technical solution, the product is a drug or experimental reagent.

[0013] The present invention also discloses a pharmaceutical composition for treating liver cancer and sensitizing the immune checkpoint inhibitor αPD-1. The pharmaceutical composition includes active ingredients, namely, the SRSF1 cleavage factor small molecule inhibitor Okanin and the immune checkpoint inhibitor αPD-1.

[0014] As a preferred technical solution, the SRSF1 inhibitor Okanin refers to the compound represented by general structural formula I;

[0015]

[0016] This invention also discloses the application of a pharmaceutical composition for the treatment of liver cancer and for enhancing the immune checkpoint inhibitor αPD-1.

[0017] This includes tumor prognosis or the preparation of tumor prognosis products;

[0018] Products that downregulate tumor glycolysis activity and enhance CD8+ T cell killing activity;

[0019] Immune checkpoint inhibitors that enhance immunity or products that prepare immune checkpoint inhibitors;

[0020] Reversing tumor immune escape or preparing products that reverse tumor immune escape;

[0021] Products that inhibit tumor resistance to immune checkpoint inhibitors.

[0022] The activity of reversing tumor drug resistance or the preparation of products that inhibit or reverse tumor drug resistance; said products are drugs or experimental reagents.

[0023] The present invention also discloses the use of a pharmaceutical composition for the treatment of liver cancer and for enhancing the immune checkpoint inhibitor αPD-1. The pharmaceutical composition treats liver cancer by improving the tumor immune microenvironment and enhancing cytotoxic CD8+ T cells.

[0024] Due to the adoption of the above-mentioned technical solutions, the application of the novel SRSF1 cleavage factor small molecule inhibitor Okanin and its drug combinations include the application of reagents for detecting SRSF1 cleavage factor expression in products for prognostic assessment of liver cancer; products that downregulate tumor cell glycolysis and enhance CD8+ T cell killing activity; products that sensitize immune checkpoint inhibitors or prepare products that sensitize immune checkpoint inhibitors; products that reverse tumor immune escape or prepare products that reverse tumor immune escape; and products that inhibit tumor resistance to immune checkpoint inhibitors or inhibit tumor resistance to immune checkpoint inhibitors.

[0025] The advantages of this invention are as follows:

[0026] This invention provides the application of Okanin targeting SRSF1 in tumor immunotherapy. Okanin directly targets the RNA-binding core region of the SRSF1 protein, preventing SRSF1 from exerting its downstream glycolytic function, thereby inhibiting tumor cell proliferation. Simultaneously, Okanin can act on CD8+ T cells, enhancing their effector function and thus increasing their activity in killing tumor cells. Furthermore, it exhibits a significant synergistic effect when used in combination with immune checkpoint inhibitors (αPD1), significantly improving the therapeutic efficacy for drug-resistant tumors. Therefore, the novel target of tumor therapy SRSF1 and its regulatory role and mechanism on immune checkpoints, as well as the anti-tumor effects of Okanin through targeting SRSF1 and sensitizing immune checkpoint inhibitors, all hold promising research potential.

[0027] In vivo and in vitro studies have demonstrated that Okanin's direct target is serine-arginine-rich splicing factor 1 (SRSF1). High expression of SRSF1 has been established as positively correlated with the prognosis of various malignant tumors, and it also inhibits the glycolytic activity of tumor cells, making it a novel target for cancer therapy. Simultaneously, Okanin enhances the cytotoxic function of CD8+ T cells, exhibiting a significant synergistic effect when used in combination with the immune checkpoint inhibitor αPD-1, significantly improving the treatment efficacy of drug-resistant tumors. Therefore, the novel cancer therapy target SRSF1, its regulatory role and mechanism in tumor glucose metabolism, and the anti-tumor effects of Okanin through targeting SRSF1 and sensitizing immune checkpoint inhibitors all hold promising research potential.

[0028] In vitro and in vivo experiments have confirmed that Okanin significantly downregulates the glycolytic activity of tumor cells while enhancing the killing function of CD8+ T cells. It has become a novel small molecule inhibitor for improving the tumor immune microenvironment and has promising applications in reversing tumor immune escape and sensitizing immune checkpoint inhibitors.

[0029] In vivo antitumor experiments showed that Okanin significantly inhibited tumor proliferation, enhanced the effector function of CD8+ T cells, and increased the number of tumor-infiltrating T lymphocytes. When used in combination with the immune checkpoint inhibitor αPD-1, it significantly inhibited the growth of drug-resistant tumors, exhibiting a good synergistic sensitization effect with low toxicity.

[0030] Computer-aided virtual screening of SRSF1 protein and in vitro molecular docking experiments revealed that Okanin mainly acts on the RRM1 region of SRSF1. Furthermore, metabolic experiments, flow cytometry, and immunohistochemistry showed that Okanin can inhibit tumor cell glucose metabolism and enhance the killing activity of CD8+ T cells. Attached Figure Description

[0031] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0032] The accompanying drawings, which form part of this specification, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. Hereinafter, embodiments of the invention will be described in detail with reference to the accompanying drawings, wherein:

[0033] Figure 1 This is a schematic diagram illustrating the expression and prognosis of SRSF1 in hepatocellular carcinoma in Example 1 of the present invention, wherein... Figure 1 A and Figure 1 B represents the expression of SRSF1 in human liver cancer and adjacent tissues. Figure 1 C represents the prognostic status of SRSF1 expression;

[0034] Figure 2 This is a schematic diagram of the virtual screening of small molecule inhibitors using SRSF1 in Example 2 of the present invention, wherein... Figure 2 A is the general formula for Okanin; Figure 2 B is a schematic diagram of the docking mode between Okanin and SRSF1 molecules; Figure 2 C represents the in vitro experimental verification of the docking between Okanin and SRSF1 molecules;

[0035] Figure 3 This is a diagram illustrating the effects of Okanin on glucose metabolism and proliferation in liver cancer cells in Example 3 of the present invention. Figure 3 A represents the effect of Okanin on the inhibition of liver cancer cell proliferation in vitro; Figure 3 B is an example of Okanin inhibiting glycolysis in tumor cells; Figure 3 In vivo experiments have shown that Okanin inhibits the proliferation of tumor cells;

[0036] Figure 4 This is a diagram illustrating the effect of Okanin in vivo on inhibiting the proliferation of liver cancer and melanoma and exhibiting a synergistic therapeutic effect with αPD-1, as shown in Example 4 of the present invention. Figure 4 A indicates that Okanin inhibits liver cancer proliferation in vivo and has a synergistic therapeutic effect with αPD-1. Figure 4 B indicates that Okanin inhibits melanoma proliferation in vivo and has a synergistic therapeutic effect with αPD-1.

[0037] Figure 5 This is a diagram illustrating the enhancement of CD8+ T cell effector function by Okanin in Example 5 of the present invention, wherein... Figure 5 A is that Okanin enhances the secretion of IFN-γ by CD8+ T cells; Figure 5 B is an Okanin-enhanced GZMB secretion by CD8+ T cells. Detailed Implementation

[0038] To overcome the above deficiencies, this invention provides the application of Okanin, a novel small molecule inhibitor of SRSF1 cleavage factor, and its drug combinations to solve the problems in the background art.

[0039] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer.

[0040] Application of Okanin, a novel small molecule inhibitor of SRSF1 cleavage factor:

[0041] a1) Tumor prognosis or preparation of tumor prognosis products;

[0042] a2) Products that downregulate tumor cell glycolysis activity or enhance CD8+ T cell killing activity;

[0043] a3) Immune checkpoint inhibitors that enhance immune sensitivity or products that prepare immune checkpoint inhibitors.

[0044] a4) Reversing tumor immune escape or preparing products that reverse tumor immune escape;

[0045] a5) Products that inhibit tumor resistance to immune checkpoint inhibitors or products that inhibit tumor resistance to immune checkpoint inhibitors.

[0046] a6) Reversing tumor drug resistance activity or preparing products that inhibit or reverse tumor drug resistance activity.

[0047] Okanin, also known as oxalis, is the active ingredient in Coreopsis grandiflora and possesses various biological activities, including antitumor and anti-inflammatory effects. This invention has revealed that Okanin can reverse resistance to immune checkpoint inhibitors with low toxicity. Furthermore, in vitro and in vivo experiments have confirmed that Okanin is a novel small-molecule inhibitor that downregulates glycolytic activity in tumor cells or enhances the killing activity of CD8+ T cells, showing promise for reversing tumor immune escape and sensitizing immune checkpoint inhibitors.

[0048] The product is a drug or experimental reagent, which can be used for basic research.

[0049] The inactive components of the drug can be pharmaceutically commonly used carriers, excipients, and diluents. Furthermore, according to conventional methods, it can be formulated into oral, topical, suppository, and sterile injectable solutions such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and sprays.

[0050] The non-pharmaceutical active ingredients that may be included, such as carriers, excipients, and diluents, are well known in the art, and those skilled in the art can determine that they meet clinical standards.

[0051] The medicament of the present invention can be administered into the body by known means. For example, it can be delivered to the tissue of interest via intravenous systemic delivery or local injection. Alternatively, it can be administered via intravenous, percutaneous, intranasal, mucosal, or other delivery methods. Such administration can be performed via single or multiple doses. It will be understood by those skilled in the art that the actual dose to be administered in the present invention can vary considerably depending on a variety of factors, such as the target cells, biological type or tissue thereof, the general condition of the subject to be treated, the route of administration, the manner of administration, etc.

[0052] The drug can be administered to humans and non-human mammals, such as mice, rats, guinea pigs, rabbits, dogs, monkeys, and chimpanzees.

[0053] The Okanin refers to the compound represented by the general structural formula (Ⅰ);

[0054]

[0055] The immune checkpoint inhibitors include PD-1 / PD-L1 inhibitors, preferably anti-PD-1 / PD-L1 antibodies.

[0056] Okanin directly targets SRSF1, binding to the RRM1 terminus of SRSF1, which contains an RNA-binding domain. It inhibits the assembly of the SRSF1 splice in tumor cells and the expression of downstream LDHA mRNA, while also increasing cytotoxic activity in CD8+ T cells.

[0057] The present invention also discloses a small molecule inhibitor that reduces the glycolytic activity of tumor cells and enhances the killing activity of CD8+ T cells, namely Okanin represented by general formula (Ⅰ).

[0058] The application of the small molecule inhibitors that reduce tumor cell glycolysis activity and enhance CD8+ T cell killing activity in reversing tumor immune escape or in the preparation of products that reverse tumor immune escape and products that sensitize anti-tumor immune checkpoint inhibitors.

[0059] The present invention also discloses a pharmaceutical composition comprising at least a target SRSF1 and an immune checkpoint inhibitor.

[0060] The substance that targets SRSF1 is Okanin.

[0061] The present invention also discloses the application of the pharmaceutical composition:

[0062] a1) Tumor prognosis or preparation of tumor prognosis products;

[0063] a2) Products that downregulate tumor cell glycolysis activity or enhance CD8+ T cell killing activity;

[0064] a3) Immune checkpoint inhibitors that enhance immune sensitivity or products that prepare immune checkpoint inhibitors.

[0065] a4) Reversing tumor immune escape or preparing products that reverse tumor immune escape;

[0066] a5) Products that inhibit tumor resistance to immune checkpoint inhibitors or products that inhibit tumor resistance to immune checkpoint inhibitors.

[0067] a6) Reversing tumor drug resistance activity or preparing products that inhibit or reverse tumor drug resistance activity.

[0068] The product is a drug or laboratory reagent.

[0069] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below with reference to specific embodiments.

[0070] Experiment 1: Expression of SRSF1 in hepatocellular carcinoma and adjacent normal tissues and its prognostic value for hepatocellular carcinoma patients

[0071] Methods: Approximately 240 clinical specimens were collected, with liver cancer tissue samples obtained from Zhongshan Hospital affiliated with Fudan University. SRSF1 expression was detected using immunohistochemistry. The tissues were fixed overnight in 10% neutral formalin, dehydrated, and then embedded in low-melting-point paraffin.

[0072] Serial 4 μm thick tissue sections were excised and fixed onto silicated slides. Immunohistochemical staining was performed using an antibiotic-streptoxin-peroxidase conjugation method. Each tissue section was deparaffinized, rehydrated, immersed in antigen retrieval solution, boiled at 100°C for 10 min, and then incubated with a peroxidase inhibitor. After blocking nonspecific binding with normal goat serum, the tissue sections were incubated overnight at 4°C with the following primary antibody: anti-SRSF1 (No. sc-33652, Santa Cruz); all antibodies were used at a 1:50 dilution. Finally, based on the corresponding clinical data and prognostic outcomes of the specimens, the relationship between SRSF1 expression and clinicopathological features and the prognosis of hepatocellular carcinoma patients was analyzed.

[0073] Statistical methods: Results from at least three independent experiments are presented as mean ± standard deviation. Statistical analysis was performed using GraphPadPrism software (version 8.0). When analyzing only two groups, two Student t-tests were used to assess differences between groups; when comparing more than two groups, one-way ANOVA was used. The Gehan-Breslow-Wilcoxon test was performed to analyze the correlation between HNRNPM protein expression and overall survival.

[0074] Result: As Figure 1As shown in Figure A, SRSF1 expression in human liver cancer is significantly higher than that in adjacent normal cells; Figure 1 High expression of SRSF1 in C indicates a poor prognosis for liver cancer.

[0075] Experiment 2: Development of small molecule inhibitors of SRSF1

[0076] Methods: First, a homology model of the SRSF1 protein was established to predict its three-dimensional structure. The first inhibitor targeting SRSF1 was searched from 20,997 small molecule compounds in the Drugbank database. The predicted structure of SRSF1 was constructed, and virtual high-throughput screening was performed to screen for potential candidate pharmacological chemicals.

[0077] result: Figure 2 A is the general formula for Okanin; Figure 2 B shows a schematic diagram of the docking mode between Okanin and SRSF1 molecules; Figure 2 C is an in vitro experimental verification that Okanin has a high binding affinity (KD) for recombinant SRSF1, with a dissociation constant of 1.37e-5M; Figure 2 In vitro experiments showed that Okanin has good inhibitory activity against SRSF1 activity, with an IC50 of approximately 144 μm.

[0078] Experiment 3: Effects of Okanin on the proliferation and metabolism of liver cancer cells in vivo and in vitro

[0079] Methods: First, cells were infected with the lentiviral vector (U6-sh-SRSF1-EGFP-IRES-puromycin) purchased from Shanghai Jikai Gene Technology Co., Ltd. to establish an SRSF1 knockdown cell line. HCC cells were infected with a recombinant lentiviral transduction unit using 0.8 μg / mL polyethylene glycol. After two weeks, stable cells were selected using 2 μg / mL puromycin. The efficiency of RNA interference was then assessed by qRT-PCR and Western blotting.

[0080] Cell viability and colony formation assays: Cell viability was determined using a Cell Counting Kit-8 (CCK-8; KeyGEN BioTECH, Jiangsu, China). HCC cells were seeded at a density of 3 × 10³ cells per well in 96-well plates, using 100 μL of cell suspension per well. After 72 h of culture, absorbance was measured at 490 nm using a microplate reader. Each experiment was performed six times, for a total of three independent experiments. The mean percentage of cell viability was then calculated. For colony formation assays, HCC cells were seeded at a density of 500 cells per well in 6-well plates and cultured in complete growth medium until visible colonies formed (approximately two weeks). Cell colonies were fixed using tissue fixative, stained with 1% crystal violet, and counted. All experiments were performed three times independently.

[0081] In vivo animal experiments: All animal experiments were conducted in accordance with the "Guideline for the Care and Use of Laboratory Animals" (NIH Publication 80-23, revised in 1996) and the "Ethical Guidelines for Animal Laboratory Facilities" formulated by Fudan University. Animals were housed in the laboratory animal facility of Zhongshan Hospital affiliated with Fudan University under pathogen-free conditions. Four-week-old female C57BL / 6 mice were subcutaneously injected with Hep1-6 liver cancer cells. 1*10^5 cells per mouse were injected into the left axilla. When the tumor reached 50 mm3, the mice were divided into two groups of six. Six days later, the mice were administered 10 mg / kg / day intraperitoneally for one week. The size of the subcutaneous tumors was observed and measured two weeks after the subcutaneous tumor cell injection to evaluate the effect of the SRSF1 inhibitor Okanin on HCC growth in mice.

[0082] Glycolysis experiment: 2 x 10 4 Hepatocellular carcinoma control and Okanin-pretreated hepatocellular carcinoma cells were seeded in XF96 culture plates and stabilized overnight. Extracellular acidification rate (ECAR) was measured using an XF96 extracellular flow analyzer and a glucose stress fuel elasticity assay kit (Agilent). ECAR measurements were performed according to the manufacturer's instructions. Results were analyzed using Wave software (Seahorse / Agilent). For Seahorse analysis using Okanin treatment, hepatocellular carcinoma cells were pretreated with 500 μM Okanin for 48 hours before the above ECAR assay was performed.

[0083] result: Figure 3 A is a CCK8 experiment demonstrating that Okanin inhibits the proliferation of liver cancer cells in vitro; Figure 3 B is a clonogenic assay demonstrating that Okanin inhibits the proliferation of tumor cells; Figure 3 C represents Okanin's inhibition of glycolysis in tumor cells; Figure 3 In vivo experiments have shown that Okanin inhibits the proliferation of tumor cells.

[0084] Experiment 4: Exploring the effects of Okanin in combination with immune checkpoint inhibitors in mice

[0085] Methods: Mouse-derived hepatocellular carcinoma and melanoma cells were inoculated into the left axilla of mice at a rate of 1*10^5 per mouse. When the tumor reached 50 mm3, the mice were randomly divided into 8 groups (4 groups of hepatocellular carcinoma cells and 4 groups of melanoma cells), with 6 mice in each group (grouping as follows: Ctrl+IgG, Ctrl+αPD-1, Okanin+IgG, Okanin+αPD-1). The dosage was Okanin (5 mg / kg) and αPD-1 (5 mg / kg). The administration schedule was: Okanin once daily for one week, and αPD-1 once every 3 days, for a total of 13 days. After this period, the mice were treated, and the tumors were harvested for subsequent analysis.

[0086] result: Figure 4 A indicates that Okanin inhibits liver cancer proliferation in vivo and has a synergistic therapeutic effect with αPD-1. Figure 4 B indicates that Okanin inhibits melanoma proliferation in vivo and has a synergistic therapeutic effect with αPD-1.

[0087] Experiment 5: Isolation and detection of tumor-infiltrating T lymphocytes

[0088] Methods: First, CD8+ T cells were positively sorted from mouse spleens using CD8 magnetic beads and randomly divided into two groups: a control group and a group pretreated with Okanin (5 μM) for 16 hours. The CD8+ T cells from both groups were then analyzed by flow cytometry, using flow cytometry antibodies to label CD8, IFN-γ, and GZMB. In the in vivo experiment, mice were euthanized by dislocation, and tumor tissue was dissected, photographed, and weighed. The tumor tissue was then cut into 2-4 mm fragments and dissected into single-cell suspensions using Miltenyi tumor tissue dissociation fluid and an eight-channel tissue processor for subsequent analysis. The dissected tumor single-cell suspensions were then analyzed by flow cytometry using antibodies labeled with CD45, CD8, IFN-γ, and GZMB.

[0089] result: Figure 5 A is that Okanin enhances the secretion of IFN-γ by CD8+ T cells; Figure 5 B enhances the secretion of GZMB by CD8+ T cells using Okanin; in different mouse liver cancers ( Figure 5 C) and melanoma ( Figure 5 D) In ​​the subcutaneous tumor group, Okanin group and Okanin+αPD-1 significantly increased IFN-γ+ and GZMB+CD8+ T cells.

[0090] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. The use of a novel SRSF1 splicing factor small molecule inhibitor Okanin in the preparation of a medicament, characterized in that, The drug is used for: a) treating hepatocellular carcinoma and / or melanoma; b) sensitizing immune checkpoint inhibitor aPD-1 for treating hepatocellular carcinoma and / or melanoma; c) reversing the drug resistance of hepatocellular carcinoma and / or melanoma to immune checkpoint inhibitor aPD-1.

2. The use of the novel SRSF1 splicing factor small molecule inhibitor Okanin according to claim 1 for the preparation of a medicament, characterized in that: The mechanism of action of the drug includes down-regulating tumor cell glycolysis and / or enhancing CD8+ T cell killing activity.

3. A pharmaceutical composition for treating liver cancer and sensitizing immune checkpoint inhibitor aPD-1, characterized in that: The pharmaceutical composition comprises active ingredients, which include SRSF1 splicing factor small molecule inhibitor Okanin and immune checkpoint inhibitor aPD-1. 4.The pharmaceutical composition for treating liver cancer and sensitizing immune checkpoint inhibitor αPD-1 of claim 3, wherein the compound is: (I). The SRSF1 inhibitor Okanin refers to a compound represented by structural formula General Formula I; General Formula I.

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