WO2022151532A1

WO2022151532A1 - Application of tegaserod maleate in treatment of acute myelocytic leukemia and colorectal cancer

Info

Publication number: WO2022151532A1
Application number: PCT/CN2021/074340
Authority: WO
Inventors: 陈璋辉; 高向伟; 洪运广
Original assignee: 陈璋辉; 高向伟; 洪运广
Priority date: 2021-01-12
Filing date: 2021-01-29
Publication date: 2022-07-21
Also published as: CN114762685A; ZA202304761B

Abstract

The present invention relates to the field of molecular biology, and in particular to tumor treatment. Specifically, tegaserod maleate obtained by screening from drugs disclosed in the prior art has an effect of treating acute myelocytic leukemia and colorectal cancer; cytological experiments and animal experiments prove that tegaserod maleate can inhibit the proliferation of acute myelocytic leukemia cell lines and human colorectal cancer cell lines, induce apoptosis and block a cell cycle in a G1 phase; in a mouse model, the survival time of a mouse can be significantly prolonged and the progression of leukemia can be blocked; moreover, it is also found for the first time that by inhibiting the function of m6A recognition protein YTHDF1 in a tumor cell, i.e., blocking the binding of YTHDF1 to an RNA containing an m6A modification, tegaserod maleate can inhibit a translation capability of a downstream key target gene to achieve the described effect.

Description

The application of tegaserod maleate in the treatment of acute myeloid leukemia and colorectal cancer

technical field

The invention belongs to the field of molecular biology, in particular to the field of tumor treatment, and relates to a therapeutic drug obtained by screening acute myeloid leukemia and colorectal cancer specific and highly expressed markers. Specifically, the present invention uses the N6-methyladenosine (m6A) recognition protein YTHDF1 for the first time to screen drugs for treating acute myeloid leukemia and colorectal cancer. Specifically, the drug is Tegaserod maleate ).

Background technique

Leukemia is a malignant clonal disease of the hematopoietic system. According to the degree of differentiation and maturation of leukemia cells and the natural process, leukemia is divided into two categories: acute and chronic. Secondly, according to the main involved cell series, it can be divided into acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML) and chronic lymphocytic leukemia (CLL) and rare types of leukemia, such as : Hairy cell leukemia, prolymphocytic leukemia, etc. The incidence rate in my country is (3-4)/100,000. In malignant tumor mortality, leukemia ranks 6th (male) and 7th (female), and in children and adults under 35 years old, it ranks 1st. The incidence of leukemia in my country is similar to that of Asian countries, but lower than that of European and American countries. The incidence rate in urban is higher than that in rural areas, AML is more common in adults, the incidence in men is higher than that in women (1.81:1), and ALL is more common in children. Currently, AML is cured in 35 to 40 percent of adult patients under the age of 60 and in 5 to 15 percent of patients over the age of 60. Most patients with AML will experience disease relapse within 3 years of diagnosis.

Colorectal cancer (CRC) is a gastrointestinal malignancy that seriously threatens human health, and its morbidity and mortality are in the forefront. Among them, the morbidity and mortality of CRC in the United States are ranked third in malignant tumors, while the incidence and mortality of CRC in my country are the fifth. Early-stage CRC can be surgically removed, and the treatment effect is good. The 5-year survival rate of patients can reach more than 90%. However, there is currently no effective treatment for metastatic CRC, and the 5-year survival rate of patients is only about 10%. The occurrence and development process of colorectal cancer in the body is very complicated, and comprehensive treatment is mostly used in clinical practice. The effect of chemotherapy for colorectal cancer is greatly affected by individual differences of patients, but postoperative chemotherapy still has the significance to improve the prognosis, and may be the only treatment that many patients can get. Drug sensitivity and reducing ineffective treatment are of great significance to improving the overall survival rate of patients.

Recent studies have pointed to the role of mRNA methylation in the regulation of gene expression. In eukaryotic cells, an abundant and conserved mRNA modification is methylation of adenosine at the N-6 position, N6-methyladenosine (m6A). m6A is the most abundant methylation modification of mRNAs in eukaryotic cells. It affects every process of the RNA life cycle, such as mRNA stability, splicing, and precursor miRNA processing. It is also involved in tissue development, stem cell repair and differentiation. , DNA damage response, biological rhythm regulation and innate immune regulation and other biological processes. The homeostasis of m6A in cells is maintained by methyltransferase complexes (METTL3, METTL14, and WTAP, etc.) and demethylases (FTO and ALKBH5), and m6A recognition proteins (YTHDF1/2 and eIF3, etc.) are specific Identify m6A sites and transmit information, thus constructing an efficient and orderly m6A regulatory network. Studies have determined that FTO and ALKBH5 mediate this reversible removal of methylation. In addition, YTH domain family proteins are major mA-binding proteins that regulate various RNA metabolisms, including mRNA splicing, degradation, and translation. YTHDF1, also known as YTH domain-containing family protein 1 or C20orf21, is a 559 amino acid protein that is localized in the cytoplasm. There is evidence that m6A-dependent mRNA regulation is essential in mammals and that defects in m6A methylation affect a variety of biological processes. M6A mRNA methylation is emerging as a pathway that affects cancer initiation and progression in a variety of tumors. So far, many studies have confirmed the functional importance of m6A modifications in leukemia, such as METTL3, METTL14, FTO and YTHDF2. However, the role of YTHDF1 protein in the development of AML and CRC remains to be explored. Screening drugs targeting YTHDF1 protein will provide a new entry point for the treatment of AML and CRC.

Tegaserod maleate, trade name Zemaco, Changronin, is a chemical. Chemical name (R)-3-(5-methoxy-1H-indole-3-methylene)-N-pentyl-iminoguanidine maleate, molecular formula is C ₂₀ H ₂₇ N ₅ O ₅ , the molecular weight is 417.46. Tegaserod maleate is a selective, partial agonist of 5-HT ₄ receptors, which is mainly used for short-term treatment of symptom relief in female patients with constipation-predominant irritable bowel syndrome.

At present, the mechanism of action of tegaserod maleate in the gastrointestinal tract is promoted, that is, tegaserod maleate is a partial agonist of indole-type selective 5-HT ₄ receptors, which stimulates the gastrointestinal tract by stimulating ₅ -HT4 receptors stimulate gastrointestinal peristaltic reflex and intestinal secretion, and inhibit visceral sensitivity. However, whether tegaserod maleate can target m6A recognition protein YTHDF1 to treat acute myeloid leukemia and colorectal cancer, and achieve the effect of delaying the progression of acute myeloid leukemia and colorectal cancer and improving the survival of patients in the prior art. Not mentioned, and there is no animal experiment to prove its therapeutic effect.

SUMMARY OF THE INVENTION

One aspect of the present invention is to provide the application of tegaserod maleate in preparing a preparation that blocks the binding of YTHDF1 to RNA containing m6A modification, thereby regulating the expression of downstream key target genes; in a specific embodiment, the wherein the key gene is CCNE2; in another specific embodiment, the regulation is inhibiting expression.

Another aspect of the present invention is to provide the use of tegaserod maleate or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating tumors, characterized in that the tegaserod maleate blocks YTHDF1 by blocking the Binding with m6A-modified RNA to regulate the translation of downstream key target genes to achieve the effect of treating tumors.

In a specific embodiment, the tumor includes but is not limited to: the tumor in this application is selected from adenocortical carcinoma, anal carcinoma, anorectal carcinoma, anal canal carcinoma, appendix carcinoma, cerebellar astrocytoma , brain astrocytoma, basal cell carcinoma, skin cancer (non-melanoma), biliary tract cancer, extrahepatic cholangiocarcinoma, intrahepatic cholangiocarcinoma, bladder cancer, bone and joint cancer, osteosarcoma, malignant fibrous histiocytoma, brain cancer , brain tumor, brain stem glioma, ependymoma, medulloblastoma, visual pathway and hypothalamic glioma, breast cancer, bronchial adenoma, nervous system cancer, nervous system lymphoma, central nervous system Cancer, Central Nervous System Lymphoma, Cervical Cancer, Chronic Lymphocytic Leukemia, Chronic Myeloid Leukemia, Chronic Myeloproliferative Disorders, Colon Cancer, Colorectal Cancer, Cutaneous T-cell Lymphoma, Lymphoma, Mycosis Fungoides, Sezary Syndrome symptoms, endometrial cancer, esophageal cancer, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, intraocular melanoma, retinoblastoma, gallbladder cancer, gastric cancer, gastrointestinal carcinoid, intergastrointestinal GIST Lineage leukemia, hairy cell leukemia, lip and oral cavity cancer, liver cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, non-Hodgkin lymphoma, primary central nervous system lymphoma, Waldenstroem macroglobulinemia, melanin tumor, mesothelioma, metastatic squamous cell carcinoma, tongue cancer, multiple endocrine neoplasia syndrome, myelodysplastic syndrome, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, oropharyngeal carcinoma, ovarian cancer, ovarian epithelial Cancer, ovarian tumors of low malignant potential, pancreatic cancer, islet cell pancreatic cancer, sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal tumor, pituitary tumor, plasma cell tumor, pleura Pulmonary blastoma, prostate cancer, rectal cancer, renal pelvis and ureteral transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, Ewing family sarcoma, Kaposi's sarcoma, synovial sarcoma, uterine cancer, uterine sarcoma, small bowel cancer , soft tissue sarcoma, squamous cell carcinoma, supratentorial primitive neuroectodermal tumor, testicular cancer, throat cancer, thymoma, urethral cancer, endometriosis, vaginal cancer, vulvar cancer, or Wilm's tumor.

In a specific embodiment, the tumor is a hematological tumor, preferably leukemia, more preferably acute myeloid leukemia.

In another specific embodiment, the tumor is a solid tumor, preferably colorectal cancer.

In another specific embodiment, the pharmaceutically acceptable salt is selected from mesylate, maleate, tartrate, succinate, acetate, difluoroacetate, fumaric acid Salts, citrates, citrates, benzenesulfonates, benzoates, naphthalenesulfonates, lactates, malates, hydrochlorides, hydrobromides, sulfates, and phosphates.

In another specific embodiment, the medicament further includes a pharmaceutically acceptable carrier, specifically, the "pharmaceutically acceptable carrier" includes any and all solvents that are physiologically compatible, Dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. In one embodiment, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, intraperitoneal, spinal or epidermal administration (eg, by injection or infusion), and the pharmaceutical compositions of the present application may include one or more Pharmaceutically acceptable salts, antioxidants, aqueous and non-aqueous carriers, and/or adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents.

In another specific embodiment, the mode of administration of the drug includes, but is not limited to, intravenous, intramuscular, subcutaneous, parenteral, intraperitoneal, spinal or epidermal administration of the drug.

In another embodiment, the medicament further comprises a buffer, a stabilizer, and optionally a surfactant. The buffer can be selected from one or more of acetate, citrate, succinate, and phosphate. Stabilizers may be selected from sugars or amino acids, preferably disaccharides such as sucrose, lactose, trehalose, maltose. Surfactant is selected from polyoxyethylene hydrogenated castor oil, glycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, preferably described polyoxyethylene sorbitan fatty acid ester is polysorbate 20,

In another specific embodiment, the dosage of the drug is 1-20 mg/kg, preferably 2-15 mg/kg, more preferably 3-10 mg/kg, most preferably 5 mg/kg.

Another aspect of the present invention is to provide the use of tegaserod maleate in the preparation of a preparation for blocking the binding reaction of the YTH domain with m6A-modified RNA, wherein the use is for non-therapeutic purposes.

Another aspect of the present invention is the use of YTHDF1 as a target for screening tumor drugs.

In another specific embodiment, the tumor is a solid tumor, preferably a gastrointestinal tumor, more preferably a colorectal cancer.

The beneficial effects of the present invention include 1) using a marker protein of acute myeloid leukemia and colorectal cancer (YTHDF1) as a target for screening drugs for treating acute myeloid leukemia and colorectal cancer, specifically, using biological information Bioinformatics technology, which simulates the high-level structure of the marker protein, and screens the drugs that can specifically bind to it from the marketed therapeutic drugs; further, the bioinformatics technology refers to virtual screening technology; further, the said bioinformatics technology refers to the virtual screening technology; Drugs are chemical drugs.

2) The present invention finds for the first time that the drug tegaserod maleate for female constipation-type irritable bowel syndrome patients can be applied to the treatment of tumors, especially acute myeloid leukemia;

3) The present invention found that tegaserod maleate is achieved by inhibiting the function of m6A recognition protein YTHDF1 in cells, thereby inhibiting cell proliferation, arresting cell cycle in G1 phase, and promoting cell apoptosis. tumor inhibition.

Description of drawings

Figure 1 Tegaserod maleate is cytotoxic to acute myeloid leukemia THP-1 cells in a dose-dependent manner, with an IC50 value of 3.58 μM (Figure 1a), inhibiting cell proliferation (Figure 1b) and promoting cell apoptosis apoptosis (Fig. 1e), arresting the cell cycle in G1 phase (Fig. 1f). Similarly, tegaserod maleate was also cytotoxic to human colorectal cancer cells HCT116 in a dose-dependent manner, with an IC50 value of 2.349 μM (Fig. 1c), and inhibited cell proliferation (Fig. 1d).

Fig. 2 Tegaserod maleate blocked the binding of m6A recognition protein YTHDF1 to m6A-modified RNAs with a K _D value of 0.7993 μM (Fig. 2a). Tegaserod maleate was administered to THP-1 cells, and the expressions of YTHDF1 and CCNE2 were detected respectively. Taking DMSO as a control, administration of tegaserod maleate did not change the protein level of YTHDF1, while the protein expression level of CCNE2 was down-regulated ( Figure 2b).

Fig. 3 In a mouse model of AML established by xenografting human THP-1 cells using severely immunodeficient NCG mice, administration of tegaserod maleate can prolong the survival time of mice and hinder the progression of leukemia.

Detailed ways

experimental method

1. Materials

Acute myeloid leukemia cell line THP-1 and human colorectal cancer cell line HCT116 were provided by Stem Cell Bank of Chinese Academy of Sciences; RPMI1640 and DMEM were purchased from Gibco Company; fetal bovine serum was purchased from Hyclone Company. Cell CountingKit-8 reagent was purchased from DOJINDO Company, hCD45-FITC antibody, hCD117-PE-Cy7 antibody, apoptosis kit and cell cycle reagent were purchased from eBioscience Company. YTHDF1 and CCNE2 antibodies were purchased from Proteintech and CST, respectively. The primers were synthesized by Hangzhou Youkang Biological Company. Tegaserod maleate was purchased from MCE Company (HY-14153A); other drugs were of domestic analytical grade. Wright-Giemsa staining solution was purchased from BASO Company; Severe immunodeficiency NCG mice were purchased from Jiangsu JiCui Yaokang Biological Company.

2. Cell culture

Acute myeloid leukemia cell lines were grown in suspension in RPMI 1640 medium containing 10% fetal bovine serum, and human colorectal cancer cell lines were adherently grown in DMEM medium containing 10% fetal bovine serum at 37°C, 5% Cultured in a CO2 humidified incubator and passaged every other day.

3. CCK-8 cell proliferation experiment:

Cell proliferation experiments were performed using DOJINDO's Cell Proliferation and Cytotoxicity Detection Kit (CCK-8):

(1) Take 10,000 THP-1 cells/well and 20,000 HCT116 cells/well in logarithmic growth phase and inoculate them in a 96-well plate, add culture medium containing different drug concentrations to each well to a final volume of 100 μl; set at 37°C, 5% CO2 culture; 3 duplicate wells and 3 control wells (blank control without cells and only culture medium).

(2) After the cells were inoculated, the time was 0 h. After 0 h, 12 h, 24 h and 72 h, 10 μl of CCK-8 solution was added to the experimental wells and incubated at 37°C for 4 h.

(3) The absorbance at the wavelength of 450 nm was measured by an enzyme-linked immunosorbent monitor, and the cell growth curve was drawn with the time as the abscissa and the absorbance value as the ordinate.

4. Apoptosis assay

(1) Take the cells in the logarithmic growth phase and inoculate them in a 6-well plate, add culture medium containing different drug concentrations to each well to a final volume of 3ml; set it at 37°C, 5% CO2 for culture; and set up 3 duplicate wells, 3 Control wells (DMSO).

(2) Collect cells, resuspend 100 μl of cell suspension in a flow tube, add Annexin V-APC and Propidium iodide flow antibodies respectively, and incubate at room temperature for 15 minutes in the dark.

(3) Cell apoptosis was detected by flow cytometry, and the ratio of cell apoptosis was compared with DMSO group as control.

5. Cell cycle experiments

(2) Collect cells, take 1×10 ⁶ cells/tube, wash twice with PBS, and centrifuge for 5 minutes at 1000 rpm each time.

(3) Remove the supernatant, add 1 ml of 70% ethanol pre-cooled at -20°C, shake and mix well, and store the specimen at -20°C overnight.

(4) Take out the sample, centrifuge at 1000 rpm for 5 minutes, remove the supernatant, wash once with PBS, and centrifuge at 1000 rpm for 5 minutes.

(5) Remove the supernatant, add the staining solution, which is composed of PI (40μg/ml) diluted with PBS, RNase A, DNase and protease-free (0.2mg/ml), mix well, and incubate in the dark for 15min.

(6) The cell cycle was detected by flow cytometry.

6. Immunoblotting

Proteins were separated by SDS-PAGE, transferred to membrane, blocked, incubated with primary antibody overnight, and incubated with horseradish peroxidase (HRP)-labeled secondary antibody at room temperature for 1 hour. Substrate ECL was added to develop color, and the results were obtained by scanning with a chemiluminescence imager.

7. Wright-Giemsa staining of bone marrow (BM) cells

Bone marrow cells were stained with Wright-Giemsa staining kit from BASO.

(1) Separate the femur of the mouse, remove the metaphysis at both ends, rinse the bone marrow cavity with 400 μl PBS repeatedly, collect the cells in a 1.5 ml Ep tube, pipette 10 μl bone marrow dropwise onto a glass slide, and smear with another glass slide ;

(2) Add 0.5ml to 1ml of A solution dropwise on the dry blood smear, and dye for 1 minute (Note: A solution is wet and covered throughout, do not let A solution dye dry);

(3) Add drop B on solution A (the drop amount is 2 to 3 times that of solution A), blow air with ear-washing balls or shake the glass slide gently, mix thoroughly for two nights, and stain for 3-10 minutes;

(4) Wash with water (the dye should not be poured out first when rinsing, it should be washed with running water to prevent the dye residue from re-applying on the specimen), let it dry, and then perform microscopic examination.

8. AML tumor burden detection experiment

The femurs of mice were isolated, bone marrow cells were collected, anti-human CD45 and CD177 antibodies were added, and the cells were incubated for 30 minutes. The red blood cells were lysed, washed twice with PBS, and the tumor load of the mice was detected by flow cytometry.

9. Xenotransplantation of human AML cells to construct an AML mouse model

1×10 ⁶ cells were collected and resuspended in 200 μl of PBS, and injected into 8-10-week-old male severely immunodeficient NCG mice through the tail vein aseptically. After the successful engraftment of leukemia cells was confirmed by flow cytometry, the drug was administered every other day. And weigh the weight of the mice to observe the life status of the mice.

10. SPR detection-SA chip

The binding affinity of m6A- and non-m6A-containing RNAs to YTHDF1 protein was assessed by SPR experiments using a Biacore T200 instrument (GE Healthcare). Serial concentrations of YTHDF1 protein were injected into the flow system and analyzed separately. Next, a series of concentrations of compounds were injected into the flow system and analyzed for 90 s, with dissociation for 120 s. The association time was set to 120s, while the dissociation time was set to 360s. After dissociation, the chip surface was regenerated by 50 mM NaOH and 1 M NaCl. Before analysis, two reference subtractions were performed to remove changes in bulk refractive index, injected noise and data drift. Binding affinity was determined by ensemble fitting to the Langmuir 1:1 binding model in Biacore evaluation software (GE Healthcare).

Example 1 Drug Screening

First, the crystal structure PDB ID: 4RCJ of hYTHDF1 was obtained from the PDB database (http://www.rcsb.org). The m6A binding region of hYTHDF1 protein was determined as the docking pocket for virtual screening, and the software used for virtual screening was

Maestro 11.4. Use the Protein Preparation Wizard Panel module to process the protein: dewatering, hydrogenation, structure optimization, deletion of redundant chains, energy minimization, etc., using the m6A binding region of hYTHDF1 protein as a docking pocket, and using the Receptor Grid Generation module to make grid files, boxes size is

Then, prepare the compounds: The 2D format of the FDA-Approved Drug Library (containing 1805 compounds) is processed through the LigPrep Module module to output the 3D structure. Finally, virtual screening: Virtual Screening Workflow module is used for virtual screening, the prepared compounds are imported, and the Glide module is used for molecular docking, that is, the receptor and ligand molecules are docked with each other through geometric matching and energy matching. Screening was performed using the high precision (XP) mode in the Glide module to obtain the ranking of small molecule compounds. Finally, manual selection is carried out, that is, the binding force between the target and the compound, the structure of the compound, etc. are manually checked, and the top 21 compounds in the FDA-Approved Drug Library are selected for output, and the inhibitory effect of the selected compounds is confirmed by cell biology experiments.

Finally, the marketed drugs were screened against the marker protein of acute myeloid leukemia (YTHDF1), and it was found that Tegaserod maleate could inhibit the function of YTHDF1 protein, thereby inhibiting its function in acute myeloid leukemia and colorectal cancer. role in cancer.

Embodiment 2 Cytological experiment of tegaserod maleate

Take well-cultured acute myeloid leukemia THP-1 cells and human colorectal cancer HCT116 cells, and administer tegaserod maleate with different concentrations to them to observe the toxicity of tegaseroate maleate to cells, and the cellular proliferation, cell cycle, and apoptosis.

First, we found that tegaserod maleate was cytotoxic through apoptosis experiments, and its IC50 for THP-1 cells and HCT116 cells were 3.58 μM and 2.349 μM, respectively (see Figure 1a, 1c); It has a concentration-dependent inhibitory effect on the proliferation of THP-1 cells and HCT116 cells (see Figure 1b, 1d);

At the same time, it can induce apoptosis of THP-1 cells and arrest the THP-1 cell cycle in G1 phase (see Figure 1c and 1d).

Example 3 The ability of tegaserod maleate to block the binding of YTHDF1 protein to RNA containing m6A modification

Based on the results of virtual screening in Example 1 and the relationship between m6A recognition protein and acute myeloid leukemia and colorectal cancer known to those skilled in the art, we predicted that tegaserod maleate would be associated with acute myeloid leukemia and colorectal cancer. The m6A recognition protein of YTHDF1 is related to YTHDF1, so we speculate that when tegaserod maleate is administered to cells, the function and key target genes of YTHDF1 can be inhibited. It can block the binding of m6A recognition protein YTHDF1 to m6A-modified RNA, and the results are shown in Figure 2a.

At the same time, we used western blotting to find that after THP-1 cells were treated with tegaserod maleate, the protein level of cyclin E2 (CCNE2) was down-regulated, while the protein level of YTHDF1 did not change, suggesting that maleic acid Tegaserod inhibited the function of YTHDF1 protein, resulting in a decrease in the translation ability of the downstream key target gene CCNE2, and achieved the purpose of hindering the progression of leukemia cells (Figure 2b).

Example 4 Xenotransplantation of human AML cells to construct an AML mouse model experiment

In order to further verify the effect of tegaserod maleate in the treatment of acute myeloid leukemia, we conducted a mouse experiment, that is, in the xenotransplantation of human AML cells to construct an AML mouse model, 5mg/kg of tegaseroate maleate significantly prolong the survival time of mice.

AML tumor burden results and bone marrow smears showed that tegaserod maleate can block the progression of acute myeloid leukemia (see Figure 3).

It is well known to those skilled in the art that the above-mentioned specific embodiments are the best implementations known in the art, but they do not limit the scope of protection of the present application. Common techniques can be included in the scope of protection claimed in this application.

Claims

The application of tegaserod maleate in the preparation of a preparation for blocking the binding of YTHDF1 to RNA containing m6A modification, thereby regulating the expression of downstream key target genes, the application is for non-therapeutic purposes.
The application according to claim 1, wherein the key gene is CCNE2, and the regulation is inhibiting expression.
The application of tegaserod maleate or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating tumors, characterized in that the tegaserod maleate blocks the binding of YTHDF1 to RNA containing m6A modification , so as to regulate the translation of downstream key target genes and achieve the effect of tumor treatment.
According to the application of claim 3, the tumor is a hematological tumor, preferably leukemia, more preferably acute myeloid leukemia; or the tumor is a solid tumor, preferably colorectal cancer.
The application described in any one of claims 3-4, the pharmaceutically acceptable salt is selected from mesylate, maleate, tartrate, succinate, acetate, difluoroacetic acid Salt, Fumarate, Citrate, Citrate, Besylate, Benzoate, Naphthalene Sulfonate, Lactate, Malate, Hydrochloride, Hydrobromide, Sulfate , and phosphates.
The use according to any one of claims 3-5, wherein the medicine further comprises a pharmaceutically acceptable carrier, preferably, the pharmaceutically acceptable carrier comprises any physiologically compatible solvent, dispersion Vehicles, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents.
The use of any one of claims 3-6, wherein the key target gene whose translation is regulated is CCNE2.
Use according to claims 3-10, wherein the administered dose is 1-20 mg/kg, preferably 2-15 mg/kg, more preferably 3-10 mg/kg, most preferably 5 mg/kg.
The use of tegaserod maleate in the preparation of a preparation for blocking the binding reaction of a YTH domain with an RNA containing m6A modification, wherein the use is for non-therapeutic purposes.
The application of YTHDF1 as a target for screening tumor drugs; preferably, the tumor is a hematological tumor or a solid tumor, preferably leukemia, more preferably acute myeloid leukemia or colorectal cancer.