Drug for treating pancreatic cancer and use thereof

Abstract

A drug for treating pancreatic cancer and use thereof. Cell experiments demonstrate that the pharmaceutical composition has a synergistic effect of inhibiting the survival of human pancreatic cancer cells and inducing the periodic inhibition and apoptosis of human pancreatic cancer cells. Moreover, anti-tumor efficacy experiments on a CDX model and clinical trials demonstrate that, compared with the prior art, the pharmaceutical combination achieves an unexpected therapeutic effect and exhibits a synergistic effect of inhibiting tumor growth. The present invention provides a better option for the treatment of pancreatic cancer.

Description

Medicament for treating pancreatic cancer and use thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a medicament for treating pancreatic cancer and use thereof. BACKGROUND

[0002] As one of the most malignant tumors of the digestive system, in recent years, the incidence of pancreatic cancer has shown a significant upward trend both at home and abroad. Surgical resection is the only effective method for pancreatic cancer patients to obtain cure opportunities and long-term survival. However, more than 80% of pancreatic cancer patients lose the opportunity for surgery due to late stage. Drug therapy can be applied to pancreatic cancer patients of all stages, including preoperative neoadjuvant / conversion therapy for resectable or borderline resectable patients, adjuvant therapy for patients after radical surgery, and treatment for locally advanced or metastatic recurrent patients. Drug therapy not only can prolong the survival time of patients, but also can relieve the pain of advanced patients and improve the quality of life.

[0003] According to the Pancreatic Cancer Diagnosis and Treatment Guidelines (2022 Edition), the neoadjuvant / conversion therapy regimen for resectable or borderline resectable pancreatic cancer mainly includes: gemcitabine + albumin-bound paclitaxel, FOLFIRINOX and mFOLFIRINOX regimen, wherein FOLFIRINOX and mFOLFIRINOX regimen use different doses of oxaliplatin, irinotecan, calcium folinate and 5-fluorouracil drug combination; the postoperative adjuvant therapy regimen is recommended to be based on gemcitabine or fluorouracil drugs (5-fluorouracil, capecitabine or tegafur); and for the treatment of unresectable locally advanced or metastatic pancreatic cancer, commonly used drugs include: gemcitabine, albumin-bound paclitaxel, 5-fluorouracil, cisplatin, oxaliplatin, irinotecan, tegafur, capecitabine and erlotinib, etc. New pancreatic cancer drugs still need to be developed to meet the clinical needs of patients.

[0004] 5-fluorouracil inhibits the synthesis of DNA by inhibiting thymidine synthetase, and has good efficacy on digestive tract tumors and other solid tumors, and is the most widely used anti-metabolic antitumor drug.

[0005] Gemcitabine is a small molecule DNA synthesis inhibitor, and its main treatment areas include tumors, digestive system diseases, endocrine and metabolic diseases, and respiratory diseases. Its approved indications include non-small cell lung cancer, peripheral T-cell lymphoma, biliary tract tumors, bladder cancer, and urothelial carcinoma.

[0006] Xiaoloni is a small molecule anti-tumor targeted drug developed and synthesized by Shenzhen Microchip Biotechnology Co., Ltd., which targets multiple protein kinases. Through high selective inhibitory activity on VEGFR / PDGFR / c-Kit, Aurora B and CSF-1R targets, it has three-pathway anti-tumor synergistic mechanism of anti-tumor angiogenesis, inhibition of tumor cell mitosis and regulation of tumor inflammatory microenvironment, and plays a comprehensive anti-tumor role. At the same time, its high target selectivity also reduces the side effect risk caused by off-target effect. The indications of Xiaoloni under research at present include soft tissue sarcoma, liver cancer, ovarian cancer, hepatocellular carcinoma, triple-negative breast cancer, etc.

[0007] Paclitaxel is a natural alkaloid with strong anti-tumor effect. Its main mechanism is to bind and stabilize microtubules, prevent tumor cell mitosis, cause cell cycle arrest in mitosis, and ultimately lead to tumor cell apoptosis or inability to continue proliferation. Paclitaxel is an early discovered microtubule stabilizer and is considered an important progress in the field of chemotherapy. Microtubules are long, filamentous and tubular protein polymers that form the main component of the cytoskeleton. Paclitaxel binds to the N-terminal 31 amino acids of the β-tubulin subunit in microtubules, which stabilizes the microtubules and increases microtubule polymerization, leading to cell death.

[0008] There is no related research and report on Xiaoloni monotherapy or its combination with other drugs for the treatment of pancreatic cancer in the prior art. SUMMARY

[0009] The technical problem solved by the present application

[0010] In view of the great demand for clinical drugs for pancreatic cancer patients with high morbidity and mortality, the purpose of the present application is to provide a new pharmaceutical composition for preventing and / or treating and / or improving pancreatic cancer. The pharmaceutical composition has achieved an unexpected therapeutic effect compared to the prior art, and the active ingredients show an unexpected synergistic effect.

[0011] SUMMARY

[0012] The first aspect of the present application provides a pharmaceutical composition for preventing and / or treating and / or improving pancreatic cancer in the first, second, third and above lines, which comprises a DNA synthesis blocker and an Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor. The DNA synthesis blocker, also known as the DNA synthesis inhibitor, refers to a substance that can participate in cell cycle DNA synthesis activity and inhibit / block DNA synthesis.

[0013] Further, the pharmaceutical composition for the first-line, second-line, third-line and above prevention and / or treatment and / or improvement of pancreatic cancer also comprises a microtubule inhibitor (or called microtubule stabilizer).

[0014] The second aspect of the present application provides the use of an Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor in the preparation of a medicament for the first-line, second-line, third-line and above prevention and / or treatment and / or improvement of pancreatic cancer in combination with a DNA synthesis blocker.

[0015] Further, the present application also provides the use of an Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor in the preparation of a medicament for the first-line, second-line, third-line and above prevention and / or treatment and / or improvement of pancreatic cancer in combination with a microtubule inhibitor and a DNA synthesis blocker.

[0016] The third aspect of the present application provides the use of an Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor in the first-line, second-line, third-line and above prevention and / or treatment and / or improvement of pancreatic cancer in combination with a DNA synthesis blocker.

[0017] Further, the present application also provides the use of an Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor in the first-line, second-line, third-line and above prevention and / or treatment and / or improvement of pancreatic cancer in combination with a microtubule inhibitor and a DNA synthesis blocker.

[0018] In some embodiments, the DNA synthesis blocker described herein is selected from a pyrimidine drug, including a uracil drug, a cytosine drug or a T thymine drug.

[0019] In some embodiments, the DNA synthesis blocker described herein is selected from a uracil drug or a cytosine drug.

[0020] In some embodiments, the DNA synthesis blocker described herein is selected from a uracil drug.

[0021] In some embodiments, the DNA synthesis blocker described herein is selected from a fluorouracil drug.

[0022] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-Fluorouracil, Tegafur, Tegafur, Cytarabine, Capecitabine, Gemcitabine, Maribavir, Fexinidazole, Miriplatin, Miriplatin Hydrate, metronidazole, Benzoylmetronidazole, Losoxantrone hydrochloride, Clofarabine, Oxaliplatin, Levofolinate Calcium, Ciclopirox, Metronidazole Hydrochloride, Secnidazole, Cisplatin, Bleomycin Sulfate, Bleomycin Hydrochloride, Azathioprine, Thiotepa, Glyfosfin, Metronidazole Disodium Phosphate, Nimorazole, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, metabolites of the foregoing.

[0023] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-Fluorouracil, Cytarabine, Gemcitabine, Tegafur, Tegafur, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, metabolites of the foregoing.

[0024] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-Fluorouracil, Cytarabine, Gemcitabine, Tegafur, Tegafur, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, metabolites of the foregoing.

[0025] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-Fluorouracil, Cytarabine, Gemcitabine, Tegafur, Tegafur, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, metabolites of the foregoing.

[0026] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-fluorouracil, tegafur, and tegafur.

[0027] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-fluorouracil, tegafur, and tegafur.

[0028] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-fluorouracil, tegafur, and tegafur.

[0029] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-fluorouracil, tegafur, and tegafur.

[0030] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-fluorouracil, tegafur, and tegafur.

[0031] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-fluorouracil, tegafur, and tegafur.

[0032] In some embodiments, the DNA synthesis blocker described herein is selected from the group consisting of 5-fluorouracil, tegafur, and tegafur.

[0033] In some embodiments, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor described herein is selected from the group consisting of: cipatinib, apatinib, bevacizumab, Barasertib (AZD1152), ZM-447439, GSK-1070916, AT9283, PHA-739358, MK0457 (VX-680), MK0475, ENMD-2076, CY-116, Hesperadin, SN-314, R763 / AS703569, BI811283, AMG 900, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, or metabolites thereof.

[0034] In some embodiments, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor described herein is selected from the group consisting of: cipatinib, apatinib, bevacizumab, Barasertib (AZD1152), ZM-447439, GSK-1070916, AT9283, PHA-739358, MK0457 (VX-680), MK0475, ENMD-2076, CY-116, Hesperadin, SN-314, R763 / AS703569, BI811283, AMG 900, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, or metabolites thereof.

[0035] In some embodiments, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor described herein is selected from the group consisting of: cipatinib, apatinib, bevacizumab, Barasertib (AZD1152), ZM-447439, GSK-1070916, AT9283, PHA-739358, MK0457 (VX-680), MK0475, ENMD-2076, CY-116, Hesperadin, SN-314, R763 / AS703569, BI811283, AMG 900, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, or metabolites thereof.

[0036] In some embodiments, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor described herein is selected from the group consisting of Sitravatinib.

[0037] In some embodiments, the microtubule inhibitor described herein is selected from the group consisting of a taxol compound or a vinblastine compound.

[0038] In some embodiments, the microtubule inhibitor described herein is selected from the group consisting of Utidelone, Docetaxel, Eribulin, Cabazitaxel, Vinflunine, Ixabepilone, paclitaxel protein-bound, BNC-105P, OXI-4503, CPC634, Ortataxel, SCB-01A, Verubulin, NSC-631570, Colchicine, Paclitaxel, Podofilox, Vindesine Sulfate, Griseofulvin, Vinblastine, Sabizabulin, Albumin-bound docetaxel, Paclitaxel poliglumex, Paclitaxel-hyaluronic acid, Paclitide, ARC-100, Crolibulin, Comprostil, Isofludelone, Fluorapacin, MAP-4343, JJH-201601, Larotaxel, ModraPac005 / r, Patupilone, Salutaxel, Aqitasai, ARC-01, BT-1769, CAP-6, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, metabolites of the foregoing.

[0039] In some embodiments, the tubulin inhibitor described herein is selected from the group consisting of Docetaxel, paclitaxel protein-bound, Colchicine, Paclitaxel, Vinblastine, Albumin-bound docetaxel, Paclitaxel poliglumex, Paclitaxel-hyaluronic acid, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, metabolites of the foregoing.

[0040] In some embodiments, the tubulin inhibitor described herein is selected from the group consisting of paclitaxel protein-bound, Paclitaxel, Paclitaxel poliglumex, Paclitaxel-hyaluronic acid.

[0041] In some embodiments, the tubulin inhibitor described herein is paclitaxel protein-bound.

[0042] In some embodiments, the pancreatic cancer described herein is locally advanced or metastatic pancreatic cancer.

[0043] In some embodiments, the pancreatic cancer described herein is pancreatic ductal adenocarcinoma.

[0044] In some embodiments, the pancreatic cancer described herein is locally advanced or metastatic pancreatic ductal adenocarcinoma.

[0045] In some embodiments, the unit dose of Xilonix described herein is about 1-100 mg, preferably about 5-80 mg, more preferably about 10-50 mg, most preferably about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg.

[0046] In some embodiments, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor described herein is selected from the group consisting of sunitinib, or a crystalline form, pharmaceutically acceptable salt thereof, a unit dose of about 1-100 mg, preferably about 5-80 mg, more preferably about 10-50 mg, most preferably about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg; in terms of sunitinib.

[0047] In some embodiments, the sunitinib is an oral formulation, preferably a tablet or capsule, for oral administration.

[0048] In some embodiments, the DNA blocking agent described herein is selected from the group consisting of 5-fluorouracil, or a crystalline form, pharmaceutically acceptable salt thereof; a unit dose of 5-fluorouracil of 10-500 mg; preferably 25-300 mg, most preferably 50 mg, 125 mg or 250 mg; in terms of 5-fluorouracil.

[0049] In some embodiments, the 5-fluorouracil is an injection formulation, for administration by injection.

[0050] In some embodiments, the 5-fluorouracil is an oral formulation, preferably a tablet, for oral administration.

[0051] In some embodiments, the DNA blocking agent described herein is selected from the group consisting of gemcitabine, or a crystalline form, pharmaceutically acceptable salt thereof; a unit dose of gemcitabine of 100-2000 mg; preferably 200-1000 mg, most preferably 200 mg or 1000 mg; in terms of gemcitabine.

[0052] In some embodiments, the gemcitabine is an injection formulation, for administration by injection.

[0053] In some embodiments, the microtubule inhibitor described herein is selected from the group consisting of paclitaxel, or a crystalline form, pharmaceutically acceptable salt thereof; a unit dose of paclitaxel of 20 mg, 50 mg, 100 mg, 500 mg, 1 g, 2 g; in terms of paclitaxel unit.

[0054] In some embodiments, the paclitaxel is an injection formulation, for administration by injection.

[0055] In some embodiments, the microtubule inhibitor is paclitaxel albumin, for administration by injection.

[0056] In some embodiments, the pharmaceutical composition as described above is used for preventing and / or treating and / or ameliorating pancreatic cancer.

[0057] The fourth aspect of the present application provides a kit comprising the pharmaceutical composition as described above.

[0058] In some embodiments, in the kit, the DNA synthesis blocker, the tubulin inhibitor and the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor are unitary formulations with the same or different specifications, and are placed in the same container or in different containers respectively.

[0059] In some embodiments, in the kit, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor is a gastrointestinal administration dosage form, preferably an oral formulation; the DNA synthesis blocker is a gastrointestinal administration dosage form or parenteral administration, preferably an oral formulation or an injection; the tubulin inhibitor is a gastrointestinal administration dosage form or parenteral administration, preferably an oral formulation or an injection; more preferably an injection.

[0060] In some embodiments, the kit further comprises instructions for use or other information.

[0061] The fifth aspect of the present application provides a method for preventing and / or treating and / or ameliorating pancreatic cancer, comprising administering to a patient in need thereof a prophylactically and / or therapeutically and / or amelioratively effective amount of the pharmaceutical composition as described above.

[0062] In some embodiments, in the method for preventing and / or treating and / or ameliorating pancreatic cancer, the DNA synthesis blocker, the tubulin inhibitor and the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-targeted kinase inhibitor are administered simultaneously, separately or sequentially.

[0063] In some embodiments, in the method for preventing and / or treating and / or ameliorating pancreatic cancer, the Xilonix is an oral formulation, preferably a capsule or a tablet, which is administered orally.

[0064] In some embodiments, in the method for preventing and / or treating and / or ameliorating pancreatic cancer, the gemcitabine is an injection, which is administered by injection.

[0065] In some embodiments, the paclitaxel is in an injectable form for administration by injection.

[0066] Definitions of terms:

[0067] In the present application, the crystal forms include non-solvated crystal forms, solvated crystal forms and amorphous structures. Among them, the crystal forms of sirolimus include non-solvated crystals A, B and C thereof. The non-solvated crystals A, B and C of sirolimus and the preparation method thereof are disclosed in CN201610856945.2, the contents of which are incorporated herein in their entirety.

[0068] In the present application, the isomers include conformational isomers, enantiomers and diastereomers.

[0069] The "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" in the present application refers to an acid addition salt or a base addition salt obtained by reacting sirolimus, paclitaxel, gemcitabine or 5-fluorouracil and a pharmaceutically acceptable acid or base. Among them, the pharmaceutically acceptable acid is preferably selected from inorganic acids (such as hydrochloric acid, sulfuric acid, phosphoric acid or hydrobromic acid, etc.), and organic acids (such as oxalic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid or benzoic acid, etc.); the pharmaceutically acceptable base is preferably selected from sodium hydroxide, potassium hydroxide, calcium hydroxide, sodium carbonate, potassium bicarbonate, ammonia or ammonium bicarbonate, etc.

[0070] The prodrug in the present application is also called precursor drug, drug precursor, precursor drug, etc., which refers to a compound obtained by chemical structural modification of a drug, which has little or no activity in vitro, and releases active drugs in vivo through enzymatic or non-enzymatic conversion to exert drug efficacy.

[0071] The metabolite in the present application refers to the intermediate metabolite and the final metabolite obtained after metabolism of the compound in the animal body.

[0072] In the present application, the "effective amount" in the prophylactic / treatment / improvement effective amount refers to a dose between the minimum limited amount and the maximum amount, which can produce obvious effects on the body without causing toxic reactions.

[0073] In the present application, the "individual" in the individual in need refers to a vertebrate. In some embodiments, the vertebrate refers to a mammal. Mammals include, but are not limited to, livestock (such as cows), pets (such as cats, dogs, and horses), primates, mice and rats. In some embodiments, the mammal refers to a human.

[0074] In the present application, the term "combination", "combined" refers to administration of two or more pharmaceutical components, which can be prepared as a combined preparation or as a combination therapy in which two or more single active ingredient preparations are administered separately. The timing of the combination depends on the combination administered. The administration of the combination is intended to include simultaneous or sequential administration, either separately or in combination. The components of the present application can be administered separately or in combination to a patient. The components described in the present application can be combined with each other, and can also be used in combination with other active agents known to be useful in the treatment of cancer.

[0075] In the present application, the term "pharmaceutical composition", "pharmaceutical combination" can be used interchangeably, which refers to two or more pharmaceutical components mixed together to prepare a combined preparation, or as a single active ingredient preparation and used in combination. When used in combination as a single active ingredient preparation, it can be administered separately or in combination to a patient.

[0076] Compared with the prior art, the present application has the following beneficial effects:

[0077] The present application proves by cell survival rate test, flow cytometry cycle detection, flow cytometry apoptosis detection, cell viability test, human pancreatic cancer CDX model anti-tumor efficacy test, etc. that the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor represented by Xioroni combined with the DNA synthesis blocker represented by 5-fluorouracil and gemcitabine can synergistically inhibit the cell survival of human pancreatic cancer cell PANC1, synergistically induce the cycle inhibition and apoptosis of human pancreatic cancer cell PANC1, inhibit the cell viability of pancreatic cancer cells, and inhibit the growth of tumors in animals. The experimental results show that the pharmaceutical composition of the present application has a synergistic effect and can well treat pancreatic cancer, providing a better choice for the treatment of pancreatic cancer.

[0078] Further, the present application also provides clinical trial data of Xioroni combined with paclitaxel and gemcitabine for treating patients with locally advanced or metastatic pancreatic ductal adenocarcinoma. The clinical trial results show that Xioroni combined with paclitaxel and gemcitabine shows good therapeutic effect in the first-line treatment of locally advanced or metastatic pancreatic ductal adenocarcinoma, with an overall response rate (ORR) of 57% (4 / 7) and a disease control rate (DCR) of 100% (7 / 7). While the existing treatment regimen of paclitaxel combined with gemcitabine has an overall response rate (ORR) of 11.11% and a DCR of 46.67% 【1】 The technical solution provided by the present application significantly improves the benefits of patients and achieves unexpected technical effects compared with the prior art, showing a synergistic effect between drugs. BRIEF DESCRIPTION OF DRAWINGS

[0079] Figure 1. The results of the cell viability inhibition test of PANC1 by Xioroni alone.

[0080] Figure 2. Results of cell viability inhibition assay of PANC1 by 5-Fluorouracil alone.

[0081] Figure 3. Results of cell viability inhibition assay of PANC1 by Xiaronil in combination with 5-Fluorouracil.

[0082] Figure 4. Results of cell cycle analysis of PANC1 by Xiaronil and / or 5-Fluorouracil.

[0083] Figure 5. Results of apoptosis induction of PANC1 by Xiaronil and / or 5-Fluorouracil.

[0084] Figure 6. Results of tumor volume change of the anti-tumor efficacy assay of SW1990 human pancreatic cancer CDX model by Xiaronil and / or Gemcitabine.

[0085] Figure 7. Results of tumor weight measurement of the anti-tumor efficacy assay of SW1990 human pancreatic cancer CDX model by Xiaronil and / or Gemcitabine.

[0086] Figure 8. Results of body weight change of the anti-tumor efficacy assay of SW1990 human pancreatic cancer CDX model by Xiaronil and / or Gemcitabine.

[0087] In the figures, Chiau. refers to Xiaronil; 5Fu refers to 5-Fluorouracil. DETAILED DESCRIPTION

[0088] The present application discloses a medicament for treating pancreatic cancer and its use, and those skilled in the art can refer to the content herein to appropriately improve the process parameters for implementation. It is particularly pointed out that all similar substitutions and changes are obvious to those skilled in the art, and they are considered to be included in the present application. The medicament for treating pancreatic cancer and its use described in the present application have been described by preferred embodiments, and relevant personnel can obviously make changes or appropriate changes and combinations to the application and pharmaceutical compositions described herein without departing from the content, spirit and scope of the present application, to realize and apply the present application technology.

[0089] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0090] Test materials

[0091] The human pancreatic cancer cell line PANC1 was purchased from the American Type Culture Collection (ATCC; CRL-1469 TM) and cultured in DMEM (Hyclone) containing 1% double antibody (penicillin-streptomycin, Hyclone) and 10% fetal bovine serum (Gibco).

[0092] Chiauranib (Chiauranib) was obtained from Shenzhen Chipscreen Biosciences Co., Ltd., and 5-fluorouracil (5FU) was purchased from MCE (Cat. No. HY-90006). Both were dissolved in dimethyl sulfoxide (DMSO; purchased from Shanghai Sangon Biotech Co., Ltd. (hereinafter referred to as "Shanghai Sangon"), Cat. No. DN3039A) to prepare a 30 mM stock solution. The solution was then stored in a -20°C refrigerator until ready for use. Working solutions of the appropriate final concentration were prepared according to the needs of each experiment.

[0093] Main experimental reagents and instruments

[0094] alamarBlue TM Cell Viability Reagent(Invitrogen,DAL1100)

[0095] Propidium iodide (PI; Shanghai Bioengineering, A601112)

[0096] Ribonuclease A (RNase A; Shanghai Bioengineering, B500474)

[0097] TritonX-100 (Shanghai Biotechnology Co., Ltd., A110694)

[0098] Annexin V-FITC / PI cell apoptosis detection kit (Yisun Biotechnology (Shanghai) Co., Ltd., 40302ES20)

[0099] Clean bench (Sujing Antai)

[0100] CO2 cell culture incubator (RS Biotech, Galaxy S)

[0101] Inverted microscope (Guangzhou Mingmei Technology Co., Ltd.)

[0102] Denovix CellDrop TM FL Cell Counter

[0103] Tecan Spark TM Multifunctional microplate reader

[0104] Refrigerated high-speed centrifuge (Eppendorf)

[0105] BD FACSCelesta TM flow cytometer

[0106] Example 1: Test the effect of drug or drug combination on cell viability

[0107] Experimental method: Collect PANC1 cells in logarithmic growth phase, inoculate 4000 per hole in 96-well cell culture plates (when testing single drug, inoculate 190 μL; when testing drug combination, inoculate 180 μL), and leave blank holes without inoculating cells and adding the same volume of culture solution; 5% CO2, 37°C culture. Incubate overnight, and then administer drugs after the cells adhere.

[0108] Before administration, first dilute sirolimus or 5FU with DMSO to 1000 times the final concentration per hole, and then dilute the DMSO dilution of sirolimus or 5FU to the culture solution at 1:50.

[0109] When testing the effect of sirolimus single drug on cell viability, add 10 μL of the corresponding DMSO dilution of sirolimus at a final concentration of 0, 1, 2, 4, 8, 16 μM per hole; when testing 5FU single drug, add 10 μL of the corresponding DMSO dilution of 5FU at a final concentration of 0, 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30 μM per hole. When testing drug combination, according to the single drug test results, add 10 μL of the corresponding DMSO dilution at a final concentration of 0, 4, 8 μM of sirolimus per hole and a final concentration of 0, 1.25, 2.5, 5 μM of 5FU per hole, so that the two drugs form different dose combinations in the cell culture plate.

[0110] After 120 hours of drug action, detect the culture solution in the 96-well culture plate, add 100 μL of detection solution (containing 90 μL of PBS and 10 μL of alamarBlue Cell Viability Reagent) per hole. Incubate at 37°C for 1-2 hours, and then read the absorbance value (Ex544 / Em590) per hole by Tecan Spark Multifunctional Enzyme Labeling Instrument. TM Cell Viability Reagent). Incubate at 37°C for 1-2 hours, and then read the absorbance value (Ex544 / Em590) per hole by Tecan Spark Multifunctional Enzyme Labeling Instrument. TM

[0111] Data processing and analysis: Each dose of drug or drug combination is repeated 3 times, the average OD BLK (background value) of the blank hole is calculated, and the OD BLK of each hole is obtained by subtracting the OD T of the blank hole from the reading of each hole. The OD T0 of the negative control hole (cells and nutrients are added to the hole without administration of drugs) is obtained. T0 The average OD T0-A of the negative control hole is calculated.The relative cell survival rate of each dosing hole was calculated according to the formula: Relative cell survival rate (%) = (OD T ÷ OD T0-A ) x 100%.

[0112] Experimental results and analysis:

[0113] The experimental results of Example 1 are shown in Figures 1-3: As shown in Figure 1, the single use of sirolimus has a certain inhibitory effect on the cell viability of human pancreatic cancer cells PANC1, and the inhibition rates of 4 μM and 8 μM doses of sirolimus on the cell viability of PANC1 are close to 20% and 50%, respectively. As shown in Figure 2, the single use of 5FU has a certain inhibitory effect on the cell viability of human pancreatic cancer cells PANC1, and a dose of 5FU within 1 μM can affect the cell viability of PANC1, and 3 μM of 5FU can produce an inhibition of close to 50% on the cell viability of PANC1. As shown in Figure 3, 4 μM and 8 μM doses of sirolimus combined with 1.25 μM, 2.5 μM and 5 μM doses of 5FU act on PANC1, and the measured cell relative viability data show that the inhibition rates of each dose of the two drug combinations on the cell viability of PANC1 are higher than those of the corresponding dose of single drug. In particular, when 4 μM dose of sirolimus is combined with 1.25 μM dose of 5FU to act on PANC1, the average inhibition rate is about 43.9%, while the inhibition rate of 4 μM dose of sirolimus single drug in this experiment is about 17.7%, and the inhibition rate of 1.25 μM dose of 5FU single drug is about 9.3%, and the inhibition effect of the combined use on PANC1 is obviously stronger than the sum of the single use of the two drugs.

[0114] In summary, the experimental results of Example 1 show that sirolimus and 5FU have a synergistic inhibitory effect on human pancreatic cancer cells PANC1.

[0115] Example 2 Flow cytometry detection of the effect of drugs or drug combinations on cell cycle

[0116] Experimental method: The PANC1 cells in logarithmic growth phase were collected by digestion, inoculated into six-well plates at 15% area per 1.8 mL per hole, and dosed after normal culture overnight.

[0117] Sirolimus or 5FU solution was prepared according to Reference Example 1 and dosed to make the final concentration of sirolimus in each hole reach 0, 4, 8 μM, respectively, and the final concentration of 5FU in each hole reach 0, 0.25, 0.5 μM, respectively, and the two drugs form different dose combinations.

[0118] After 48 hours of compound treatment, the culture medium was collected into labeled 15 mL centrifuge tubes, and the adherent cells in the wells were gently washed once with PBS, and the wash was collected into the corresponding 15 mL centrifuge tubes. 500 μL of trypsin digestion solution was added to the remaining adherent cells in the culture plate, and the plate was incubated at 37°C for 5 minutes. After the cells were fully dispersed, the digestion reaction was terminated by adding culture medium containing 10% FBS, and the resuspended cells were combined into the corresponding 15 mL centrifuge tubes. The tubes were centrifuged at 1000 rpm for 5 minutes at room temperature, the supernatant was removed, and the cells were washed with 1 mL of PBS, centrifuged at 1000 rpm for 5 minutes at 4°C, the supernatant was removed, and the cells were resuspended in 300 μL of PBS and added dropwise to 1.5 mL centrifuge tubes containing 700 μL of pre-cooled anhydrous ethanol. The tubes were gently inverted several times to mix the contents, and the cells were collected and fixed. The samples were allowed to stand at 4°C for at least 12 hours.

[0119] The PBS was mixed with a 20 mg / mL PI stock solution, a 10 mg / mL RNase A solution, and 10% Triton X-100 at a ratio of 1000:2.5:2:10 to form a DNA staining solution. The fixed cell samples were centrifuged at 1300 rpm for 10 minutes at 4°C, the supernatant was removed, and the cells were washed twice with PBS, each time by centrifuging at 1300 rpm for 10 minutes at 4°C and removing the supernatant. Each tube was resuspended in 500 μL of the above-mentioned staining solution, and the tubes were incubated at 37°C in the dark for 30 minutes. The tubes were then centrifuged at 1300 rpm for 5 minutes, the supernatant was removed, and the cells were resuspended in 200 μL of PBS and filtered through a 200-mesh stainless steel screen. The filtrate was analyzed by flow cytometry for cell cycle distribution (red fluorescence was detected at an excitation wavelength of 488 nm, and light scattering was also detected; 10,000 cells were counted for each sample).

[0120] Data processing and statistical analysis: The above-mentioned doses of drugs or drug combinations were each repeated twice, and the cell DNA content and light scattering were analyzed using FlowJo software to obtain the cycle distribution data for each sample.

[0121] Experimental results and analysis:

[0122] The experimental results of Example 2 are shown in Figure 4. Relative to the blank control, the proportion of PANC1 cells in the G2 / M phase increased slightly under the action of 4 μM of Xilonix alone; the proportion of PANC1 cells in the polyploid phase increased significantly under the action of 8 μM of Xilonix alone. Relative to the blank control, the proportion of PANC1 cells in the S phase increased slightly under the action of 0.25 and 0.5 μM of 5FU alone, which is related to the mechanism of 5FU blocking DNA synthesis (in the S phase, 5FU is incorporated into newly synthesized DNA during DNA synthesis, which blocks DNA synthesis, causing the cell cycle to stop in the S phase). When Xilonix and 5FU were combined at the above doses, the proportion of cells in the S phase increased significantly relative to 5FU alone and was significantly dose-dependent.

[0123] In summary, the experimental results of Example 2 show that Xilonix has a synergistic sensitization effect on the mechanism of action of 5FU.

[0124] Example 3: Flow cytometry for detecting the induction of apoptosis by drugs or drug combinations

[0125] Experimental method: Refer to Example 2 for cell inoculation and drug treatment, so that the final concentration of Xilonix in each well is 0 or 8 μM, the final concentration of 5FU in each well is 0 or 0.5 μM, and the two drugs form different dose combinations.

[0126] According to the Annexin V-FITC / PI Apoptosis Detection Kit Instructions, collect the cells by trypsin digestion without EDTA, centrifuge at 4°C and 300 g for 5 minutes, wash the cells twice with pre-cooled PBS, each time centrifuge at 4°C and 300 g for 5 minutes, and then remove the supernatant. Add 100 μL of 1x Binding Buffer to resuspend the cells, then add 5 μL of Annexin V-FITC and 10 μL of PI Staining Solution to each tube, and mix gently. React for 10-15 minutes at room temperature in the dark. Add 400 μL of 1x Binding Buffer, mix well, and place on ice. Then perform flow cytometry through the FL1 and FL2 channels, and analyze the apoptosis using FlowJo software.

[0127] Experimental results and analysis:

[0128] The test results of Example 3 are shown in Figure 5. The effect of 5FU monotherapy at a dose of 0.5 μM did not cause more apoptosis of human pancreatic cancer cells PANC1 relative to the blank control. The proportion of apoptotic cells in PANC1 increased from 8.18% of the background to 18.8% when XioRoNi was used alone at a dose of 8 μM. When the two drugs were used in combination, the proportion of apoptotic cells increased to 22.3%.

[0129] The experimental results of Example 3 show that although 5FU alone does not induce apoptosis of PANC1 cells, it can synergistically enhance the induction of apoptosis by XioRoNi.

[0130] Example 4 tests the effect of XioRoNi combined with gemcitabine on the viability of human pancreatic cancer cells

[0131] Test materials:

[0132] The human pancreatic cancer cell line PANC-1 was purchased from Nanjing Kebai Biotechnology Co., Ltd. (CBP60545) and cultured in DMEM medium (Gibco) containing 10% fetal bovine serum (Gibco). The human pancreatic cancer cell line MIA PaCa-2 was purchased from Wuhan Punuo Life Science and Technology Co., Ltd. (CL-0627) and cultured in MIA PaCa-2 cell-specific medium (Punuo, CM-0627, DMEM + 10% FBS + 2.5% HS + 1% P / S).

[0133] XioRoNi (Chiauranib, Chiau.) was sourced from Shenzhen Weicai Biotechnology Co., Ltd., and gemcitabine (Gemcitabine, GEM) was purchased from MCE Company (HY-17026). Both were dissolved in dimethyl sulfoxide (Dimethyl sulfoxide, DMSO; purchased from Sigma, D2650-100ML) to prepare 10 mM stock solutions, which were stored in a refrigerator at -20°C for use. The working solutions were prepared according to the requirements of each experiment.

[0134] Main experimental reagents and instruments and equipment

[0135] Luminescent Cell Viability Assay (Promega, G7572)

[0136] Tecan Spark TM Multifunctional enzyme marker

[0137] Experimental method: logarithmic growth phase PANC-1, MIA PaCa-2 cells were collected by digestion, 900 and 400 cells were inoculated in 96-well cell culture plates per well, 5% CO2, 37℃ incubated overnight, after cell adhesion, drug administration. Xioroni effect 120 hours, the final concentration is 0, 2.1, 3.2, 4.7, 7.1, 10.7, 16 μM. Seal the 96-well plate with sealing film, centrifuge at 1000 rpm for 5 minutes. Slowly aspirate the culture solution in the 96-well plate, add 90 μL complete medium per well, then add 10 μL gemcitabine for 120 hours, the final concentration is 0, 0.04, 0.08, 0.16, 0.32 μM. Each concentration is set with double duplicate wells. At the same time, set the DMSO control well (0.5%, 6 duplicate wells) with the same concentration.

[0138] The cell proliferation inhibition rate of each drug administration hole was calculated according to the formula: inhibition rate (%) = (1-(RLU T -RLU BG ) / (RLU NC -RLU BG ))×100,

[0139] Wherein, RLU T represents the luminescence value of the drug administration hole, RLU BG represents the luminescence value of the culture medium background, and RLU NC represents the luminescence value of the DMSO control well.

[0140] Experimental results and analysis:

[0141] The test results of Example 4 are shown in Tables 1-2.

[0142] Table 1 shows that when xioroni and gemcitabine are used alone to act on human pancreatic cancer cells PANC-1, they have a certain inhibitory effect on the activity of PANC-1 cells. When the two are used together, they show unexpected synergistic inhibition, and the inhibition of the activity of human pancreatic cancer cells PANC-1 by the combined regimen is significantly improved compared with single drug.

[0143] Table 1 shows the results of xioroni and gemcitabine combined with PANC-1 cell proliferation inhibition rate (%)

[0144] Table 2 shows that when xioroni and gemcitabine are used alone to act on human pancreatic cancer cells MIA PaCa-2, they have a certain inhibitory effect on the activity of MIA PaCa-2 cells. When the two are used together, they show unexpected synergistic inhibition, and the inhibition of the activity of human pancreatic cancer cells MIA PaCa-2 by the combined regimen is significantly improved compared with single drug.

[0145] Table 2 Inhibition rate of combination of Chiauranib and Gemcitabine on MIA PaCa-2 cell proliferation

[0146] In summary, the experimental results of Example 4 show that the combination of Chiauranib and Gemcitabine has an unexpected synergistic inhibitory effect on human pancreatic cancer cells PANC-1 and MIA PaCa-2, and can be used for the preparation of drugs related to pancreatic cancer.

[0147] Example 5 Test of the anti-tumor efficacy of Chiauranib combined with Gemcitabine on SW1990 human pancreatic cancer CDX model

[0148] Test materials:

[0149] The human pancreatic cancer cell line SW1990 was purchased from Nanjing CBP Biotech Co., Ltd. (CBP60691) and cultured in MEM medium (Gibco) containing 10% fetal bovine serum (Gibco), 1% NEAA (Gibco), and 1 mM sodium pyruvate (Gibco).

[0150] Chiauranib (Chiau.) was sourced from Shenzhen Wechip Biotech Co., Ltd. and administered as a suspension at a concentration of 0.5 mg / mL after being prepared in 0.2% CMC-Na aqueous solution containing 0.1% Tween 80. Gemcitabine (GEM) was purchased from MCE (HY-B0003) and administered as a solution at a concentration of 2.5 mg / mL after being prepared in PBS (Bi Yun Tian, C0221A).

[0151] Experimental method: logarithmically growing SW1990 cells were collected by digestion, counted, and then adjusted to a cell density of 4×10 7 6 / mL using 1:1 serum-free medium and Matrigel. 18-20g NOD / SCID mice (Beijing Huafukang Biotechnology Co., Ltd.) were each subcutaneously inoculated with 100μL of the cell suspension. On the 10th day after inoculation, animals with tumor volumes of 100-200mm 3 were randomly divided into 4 groups (N=7), namely the control group, the Gemcitabine monotherapy group, the Chiauranib monotherapy group, and the combination group. Gemcitabine was administered at 25mg / kg by intraperitoneal injection, once every three days; Chiauranib was administered at 5mg / kg by oral administration, once a day; the combination group was given both Gemcitabine and Chiauranib, and the dosing regimen was consistent with the monotherapy groups.

[0152] After the animals were inoculated with SW1990 cells, body weight and tumor volume were measured every Monday, Wednesday, and Friday, and tumor volume was calculated as length x width 2 x 0.5. One day after the last administration, the mice were euthanized and the tumor mass was weighed.

[0153] The tumor inhibition rate TGI of each administration group was calculated according to the formula: TGI (tumor volume) was calculated according to the following formula: TGI TV = 1 - ((V Tt -V T0 ) / (V Ct -V C0 )) x 100%.

[0154] Wherein V Tt is the tumor volume of each measurement of the treatment group, V T0 is the tumor volume measured when the treatment group is grouped; V Ct group is the tumor volume of each measurement of the control group, V C0 is the tumor volume measured when the control group is grouped;

[0155] TGI (tumor weight) was calculated according to the following formula: TGI TW = 1 - (W T / W C ) x 100%. Wherein W T is the tumor weight at the end of administration of the treatment group; W C is the tumor weight at the end of administration of the control group.

[0156] The data were statistically analyzed using GraphPad Prism 9.0 software, and the data were expressed as Mean ± SEM (standard error). Two-way comparison between each administration group and the control group was analyzed using one-way analysis of variance (ANOVA), and Brown-Forsythe method was used for variance homogeneity, and Dunnett's T3 method was used for variance heterogeneity.

[0157] Experimental results and analysis:

[0158] The test results of Example 5 are shown in Figures 6 and 7. 5 mg / kg of Xioroni was administered for 25 days, which significantly reduced the tumor volume (P < 0.01) and tumor weight (P < 0.01), and the tumor inhibition rates calculated according to the tumor volume and tumor weight were about 69% and 65%, respectively. The tumor volume and tumor weight inhibition rates of 25 mg / kg of gemcitabine were 51% and 48%, respectively. The anti-tumor effect of Xioroni combined with gemcitabine was significantly better than that of the single-drug group, and the tumor volume (P < 0.001) and tumor weight (P < 0.001) were significantly reduced compared with the single-drug group, and the tumor inhibition rates reached 90% and 82%, respectively. Xioroni combined with gemcitabine had a synergistic effect on the anti-tumor efficacy.

[0159] As shown in Figure 8, during the test period, the body weight of the mice showed an overall upward trend, and there was no significant difference in body weight between the administration group and the control group. The mice tolerated the test drug dose well.

[0160] In summary, the experimental results of Example 5 show that in the human pancreatic cancer SW1990 CDX model, Xioroni has good antitumor effect, combination with gemcitabine is effective and safe in resisting tumors, and has synergistic effect in drug efficacy.

[0161] Example 6 Single-arm, open, multi-center phase II clinical study of Xioroni combined with nab-paclitaxel and gemcitabine for first-line treatment of patients with locally advanced or metastatic pancreatic ductal adenocarcinoma

[0162] 1. Inclusion criteria: (1) Age: 18-75 years old; (2) Histologically or cytologically confirmed unresectable locally advanced or metastatic pancreatic ductal adenocarcinoma (PDAC); (3) No first-line systemic treatment; (4) ECOG PS 0 or 1.

[0163] 2. Drugs and dosing regimen: The trial uses a combination dosing regimen, and the dosing regimen of each group is as follows:

[0164] Xioroni (from Shenzhen Microchip Biotechnology Co., Ltd.): 35 mg or 50 mg, oral, qd.

[0165] Nab-paclitaxel (Ou Yi Pharmaceutical Co., Ltd.): 125 mg / m 2 , IV, D1, 8, 15 q4w.

[0166] Gemcitabine (Nanjing Zhongda Sunshine Pharmaceutical Co., Ltd.): 1000 mg / m 2 , IV, D1, 8, 15 q4w.

[0167] 3. Trial process and efficacy evaluation:

[0168] The trial is divided into two stages of dose escalation and dose expansion, of which 4 cases are enrolled in the Xioroni 35 mg group and 3 cases are enrolled in the 50 mg group in the dose escalation stage; in the dose expansion stage, a total of 28 cases are enrolled in the trial, all of which are administered according to the Xioroni 50 mg group. The main endpoint of the trial: investigator-assessed PFS (RECIST v1.1); secondary endpoints: ORR, DOR, DCR, TTR, OS, safety, PK characteristics.

[0169] Efficacy evaluation:

[0170] According to RECIST v1.1, the efficacy of tumor lesions was evaluated. As of March 25, 2025, there were 7 subjects who had at least one efficacy evaluation.

[0171] 4. Trial results

[0172] (1) DLT evaluation: During the dose escalation phase, except for one case in the 35 mg sioroni group whose chemotherapy dosage was less than 75% during the DLT observation period and was considered an unevaluable case (one case was replaced), the other six cases (three cases each in the 35 mg group and the 50 mg group, including one replaced case) completed the DLT observation and no DLT events occurred.

[0173] (2) Efficacy evaluation: As of March 25, 2025, 7 subjects had at least one efficacy evaluation, including 4 PRs (2 of which were confirmed) and 3 SDs.

[0174] Conclusion: Clinical trial results showed that sioroni combined with albumin-paclitaxel and gemcitabine showed good therapeutic effect in the first-line treatment of locally advanced or metastatic pancreatic ductal adenocarcinoma, with an overall response rate (ORR) of 57% (4 / 7) and a disease control rate (DCR) of 100% (7 / 7). The existing treatment regimen of albumin-paclitaxel combined with gemcitabine has an overall response rate (ORR) of 11.11% and a DCR of 46.67%. 【1】 The technical solution provided by the present invention significantly improves the benefits to patients and achieves unexpected technical effects compared to the existing technology, with the drugs showing a synergistic effect.

[0175] 【1】Analysis of the efficacy of gemcitabine plus albumin-bound paclitaxel combined with PD-1 monoclonal antibody in the treatment of unresectable pancreatic cancer. Yongjun Li, Guizhou Medical University, DOI: 10.27045 / d.cnki.ggyyc.2022.000411.

[0176] The present invention has been introduced in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core ideas, including the best mode, and also to enable any technician in the field to practice the present invention. It should be pointed out that for ordinary technicians in this technical field, without departing from the principles of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications also fall within the scope of protection of the claims of the present invention. The scope of patent protection of the present invention is defined by the claims and may include other embodiments that can be thought of by those skilled in the art. If these other embodiments have structural elements that are not different from the literal expression of the claims, or if they include equivalent structural elements that are not substantially different from the literal expression of the claims, then these other embodiments should also be included in the scope of the claims.

Claims

1. A pharmaceutical composition for the prevention and / or treatment and / or improvement of pancreatic cancer in first-line, second-line, third-line or higher-level settings, characterized by: Including DNA synthesis blockers and Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitors; Preferably, the DNA synthesis inhibitor is selected from pyrimidine drugs, including uracil drugs, cytosine drugs or T thymine drugs; Preferably, the DNA synthesis inhibitor is selected from uracil drugs or cytosine drugs; Preferably, the DNA synthesis inhibitor is selected from uracil drugs; Preferably, the DNA synthesis inhibitor is selected from fluorouracil drugs; Preferably, the DNA synthesis inhibitor is selected from 5-fluorouracil, tegafur, S-1, cytarabine, capecitabine, gemcitabine, maribavir, fexinidazole, miplatin, miplatin hydrate, metronidazole, metronidazole benzoate, losoxantrone, clofarabine, oxaliplatin, leucovorin calcium, ciclopirox, metronidazole hydrochloride, secnidazole, cisplatin, bleomycin sulfate, bleomycin hydrochloride, azathioprine, thiotepa, glyphosate mustard, metronidazole disodium phosphate, nimorazole, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the aforementioned drugs; Preferably, the DNA synthesis inhibitor is selected from 5-fluorouracil, cytarabine, gemcitabine, tegafur, S-1, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the aforementioned drugs; Preferably, the DNA blocking agent is selected from 5-fluorouracil, gemcitabine, or a crystal form, a pharmaceutically acceptable salt, a stereoisomer, a tautomer, a prodrug or a metabolite thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil, its crystalline form, pharmaceutically acceptable salt, stereoisomer, tautomer, prodrug or metabolite thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil, a crystalline form thereof or a pharmaceutically acceptable salt thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil; Or preferably, the DNA blocking agent is selected from gemcitabine, its crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, prodrug or metabolite thereof; Preferably, the DNA blocking agent is selected from gemcitabine, its crystalline form or its pharmaceutically acceptable salt; Preferably, the DNA synthesis inhibitor is selected from gemcitabine; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from: cioroni, apatinib, bevacizumab, barasetib (AZD1152), ZM-447439, GSK-1070916, AT9283, PHA-739358, MK0457 (VX-680), MK0475, ENMD-2076, CY-116, Hesperadin, SN-314, R763 / AS703569, BI811283, AMG 900, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs or metabolites thereof of the foregoing drugs; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, its crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs or metabolites thereof; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, a crystalline form thereof or a pharmaceutically acceptable salt thereof; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib; Preferably, the pancreatic cancer is locally advanced or metastatic pancreatic cancer; Preferably, the pancreatic cancer is pancreatic ductal adenocarcinoma; More preferably, The pharmaceutical composition further comprises a tubulin inhibitor; Preferably, the microtubule inhibitor is selected from paclitaxel compounds or vincristine compounds; Preferably, the tubulin inhibitor is selected from the group consisting of Eudetron, Docetaxel, Eribulin, Cabazitaxel, Vinflunine Tartrate, Ixabepilone, Albumin-bound Paclitaxel, BNC-105P, OXI-4503, CPC634, Ortataxel, SCB-01A, Verubulin, NSC-631570, Colchicine, Paclitaxel, Podophyllotoxin, Vindesine Sulfate, Griseofulvin, Vinblastine, Sabizabulin, Docetaxel Albumin-bound, Paclitaxel poliglumex, Paclitaxel-hyaluronic acid acid, taxane, ARC-100, Crolibulin, Comparidin disodium phosphate, Isofludelone, Flurapisin, MAP-4343, JJH-201601, larotaxel, ModraPac005 / r, Patupilone, xinalitaxel, azitaxel, ARC-01, BT-1769, CAP-6, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the foregoing drugs; Preferably, the tubulin inhibitor is selected from docetaxel, albumin-bound paclitaxel, colchicine, paclitaxel, vinblastine, albumin-bound docetaxel, paclitaxel poliglumex, paclitaxel-hyaluronic acid, and their crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites; Preferably, the tubulin inhibitor is selected from albumin-bound paclitaxel, paclitaxel, Paclitaxel poliglumex, Paclitaxel-hyaluronic acid; Preferably, the tubulin inhibitor is selected from albumin-bound paclitaxel.

2. The pharmaceutical composition according to claim 1, wherein: The Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, or a crystal form or a pharmaceutically acceptable salt thereof. The unit dose of sioronib is about 1-100 mg, preferably about 5-80 mg, more preferably about 10-50 mg, and most preferably about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg; calculated as sioronib.

3. The pharmaceutical composition according to any one of claims 1 to 2, wherein: The DNA blocking agent is selected from 5-fluorouracil, or a crystal form or a pharmaceutically acceptable salt thereof; the unit dose of the 5-fluorouracil is 10-500 mg; preferably 25-300 mg, and most preferably 50 mg, 125 mg or 250 mg, calculated as 5-fluorouracil.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein: The DNA blocking agent is selected from gemcitabine, or a crystal form or a pharmaceutically acceptable salt thereof; the unit dose of the gemcitabine is 100-2000 mg, preferably 200-1000 mg, and most preferably 200 mg or 1000 mg, calculated as gemcitabine.

5. The pharmaceutical composition according to any one of claims 1 to 4, characterized in that: The microtubule protein inhibitor is selected from paclitaxel or a crystal form or a pharmaceutically acceptable salt thereof; the unit dose of paclitaxel is 20 mg, 50 mg, 100 mg, 500 mg, 1 g, or 2 g, calculated in paclitaxel units.

6. The pharmaceutical composition according to any one of claims 1 to 5, which is used for the prevention and / or treatment and / or improvement of pancreatic cancer in the first-line, second-line, third-line or higher line settings; Preferably, the pancreatic cancer is locally advanced or metastatic pancreatic cancer; Preferably, the pancreatic cancer is pancreatic ductal adenocarcinoma.

7. A medicine box, characterized in that: A pharmaceutical composition comprising the pharmaceutical composition of any one of claims 1 to 5, wherein the DNA synthesis inhibitor, tubulin inhibitor, and Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor are unit preparations having the same or different strengths, and the DNA synthesis inhibitor, tubulin inhibitor, and Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor are placed in the same container or in different containers; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is a gastrointestinal dosage form, preferably an oral preparation; the DNA synthesis inhibitor is a gastrointestinal dosage form or parenteral administration, preferably an oral preparation or an injection; the tubulin inhibitor is a gastrointestinal dosage form or parenteral administration, preferably an oral preparation or an injection; more preferably an injection.

8. A method for preventing and / or treating and / or ameliorating pancreatic cancer in the first, second, third or higher line, comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition according to any one of claims 1 to 5; Preferably, the pancreatic cancer is locally advanced or metastatic pancreatic cancer; Preferably, the pancreatic cancer is pancreatic ductal adenocarcinoma; Preferably, the DNA synthesis inhibitor, the tubulin inhibitor and the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor are administered simultaneously, separately or sequentially.

9. Use of Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitors in combination with DNA synthesis blockers for the prevention and / or treatment and / or amelioration of pancreatic cancer as a first-line, second-line, third-line, or higher-line treatment, and / or in the preparation of drugs for the prevention and / or treatment and / or amelioration of pancreatic cancer as a first-line, second-line, third-line, or higher-line treatment, in combination with DNA synthesis blockers; Preferably, the DNA synthesis inhibitor is selected from pyrimidine drugs, including uracil drugs, cytosine drugs or T thymine drugs; Preferably, the DNA synthesis inhibitor is selected from uracil drugs or cytosine drugs; Preferably, the DNA synthesis inhibitor is selected from uracil drugs; Preferably, the DNA synthesis inhibitor is selected from fluorouracil drugs; Preferably, the DNA synthesis inhibitor is selected from 5-fluorouracil, tegafur, S-1, cytarabine, capecitabine, gemcitabine, maribavir, fexinidazole, miplatin, miplatin hydrate, metronidazole, metronidazole benzoate, losoxantrone, clofarabine, oxaliplatin, leucovorin calcium, ciclopirox, metronidazole hydrochloride, secnidazole, cisplatin, bleomycin sulfate, bleomycin hydrochloride, azathioprine, thiotepa, glyphosate mustard, metronidazole disodium phosphate, nimorazole, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the aforementioned drugs; Preferably, the DNA synthesis inhibitor is selected from 5-fluorouracil, cytarabine, gemcitabine, tegafur, S-1, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the aforementioned drugs; Preferably, the DNA blocking agent is selected from 5-fluorouracil, gemcitabine, or a crystal form, a pharmaceutically acceptable salt, a stereoisomer, a tautomer, a prodrug or a metabolite thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil, its crystalline form, pharmaceutically acceptable salt, stereoisomer, tautomer, prodrug or metabolite thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil, a crystalline form thereof or a pharmaceutically acceptable salt thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil; Or preferably, the DNA blocking agent is selected from gemcitabine, its crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, prodrug or metabolite thereof; Preferably, the DNA blocking agent is selected from gemcitabine, its crystalline form or its pharmaceutically acceptable salt; Preferably, the DNA synthesis inhibitor is selected from gemcitabine; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from: cioroni, apatinib, bevacizumab, barasetib (Barasertib, AZD1152), ZM-447439, GSK-1070916, AT9283, PHA-739358, MK0457 (VX-680), MK0475, ENMD-2076, CY-116, Hesperadin, SN-314, R763 / AS703569, BI811283, AMG 900, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs or metabolites of the aforementioned drugs. Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, its crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs or metabolites thereof; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, a crystalline form thereof or a pharmaceutically acceptable salt thereof; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, or a crystalline form or a pharmaceutically acceptable salt thereof, and the unit dose of sioronib is about 1-100 mg, preferably about 5-80 mg, more preferably about 10-50 mg, and most preferably about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg; calculated as sioronib; Preferably, the DNA blocking agent is selected from 5-fluorouracil, or a crystal form or a pharmaceutically acceptable salt thereof; the unit dose of the 5-fluorouracil is 10-500 mg; preferably 25-300 mg, and most preferably 50 mg, 125 mg or 250 mg, calculated as 5-fluorouracil; Preferably, the DNA blocking agent is selected from gemcitabine, or a crystal form or pharmaceutically acceptable salt thereof; the unit dose of gemcitabine is 100-2000 mg; preferably 200-1000 mg, and most preferably 200 mg or 1000 mg, calculated as gemcitabine; Preferably, the pancreatic cancer is locally advanced or metastatic pancreatic cancer; Preferably, the pancreatic cancer is pancreatic ductal adenocarcinoma.

10. Use of Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitors in combination with tubulin inhibitors and DNA synthesis blockers for the prevention and / or treatment and / or amelioration of pancreatic cancer as a first-line, second-line, third-line, or higher-level treatment, and / or in the preparation of medicaments for the prevention and / or treatment and / or amelioration of pancreatic cancer as a first-line, second-line, third-line, or higher-level treatment, in combination with tubulin inhibitors and DNA synthesis blockers; Preferably, the DNA synthesis inhibitor is selected from pyrimidine drugs, including uracil drugs, cytosine drugs or T thymine drugs; Preferably, the DNA synthesis inhibitor is selected from uracil drugs or cytosine drugs; Preferably, the DNA synthesis inhibitor is selected from uracil drugs; Preferably, the DNA synthesis inhibitor is selected from fluorouracil drugs; Preferably, the DNA synthesis inhibitor is selected from 5-fluorouracil, tegafur, S-1, cytarabine, capecitabine, gemcitabine, maribavir, fexinidazole, miplatin, miplatin hydrate, metronidazole, metronidazole benzoate, losoxantrone, clofarabine, oxaliplatin, leucovorin calcium, ciclopirox, metronidazole hydrochloride, secnidazole, cisplatin, bleomycin sulfate, bleomycin hydrochloride, azathioprine, thiotepa, glyphosate mustard, metronidazole disodium phosphate, nimorazole, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the aforementioned drugs; Preferably, the DNA synthesis inhibitor is selected from 5-fluorouracil, cytarabine, gemcitabine, tegafur, S-1, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the aforementioned drugs; Preferably, the DNA blocking agent is selected from 5-fluorouracil, gemcitabine, or a crystal form, a pharmaceutically acceptable salt, a stereoisomer, a tautomer, a prodrug or a metabolite thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil, its crystalline form, pharmaceutically acceptable salt, stereoisomer, tautomer, prodrug or metabolite thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil, a crystalline form thereof or a pharmaceutically acceptable salt thereof; Preferably, the DNA blocking agent is selected from 5-fluorouracil; Or preferably, the DNA blocking agent is selected from gemcitabine, its crystal form, pharmaceutically acceptable salt, stereoisomer, tautomer, prodrug or metabolite thereof; Preferably, the DNA blocking agent is selected from gemcitabine, its crystalline form or its pharmaceutically acceptable salt; Preferably, the DNA synthesis inhibitor is selected from gemcitabine; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from: cioroni, apatinib, bevacizumab, barasetib (Barasertib, AZD1152), ZM-447439, GSK-1070916, AT9283, PHA-739358, MK0457 (VX-680), MK0475, ENMD-2076, CY-116, Hesperadin, SN-314, R763 / AS703569, BI811283, AMG 900, and the crystal forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs or metabolites of the aforementioned drugs. Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, its crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs or metabolites thereof; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, a crystalline form thereof or a pharmaceutically acceptable salt thereof; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib; Preferably, the microtubule inhibitor is selected from paclitaxel compounds or vincristine compounds Preferably, the tubulin inhibitor is selected from the group consisting of Utidelon, Docetaxel, Eribulin, Cabazitaxel, Vinflunine Tartrate, Ixabepilone, Albumin-bound Paclitaxel, BNC-105P, OXI-4503, CPC634, Ortacil, SCB-01A, Verubulin, NSC-631570, Colchicine, Paclitaxel, Podophyllotoxin, Vindesine Sulfate, Griseofulvin, Vinblastine, Sabizabulin, Docetaxel Albumin-bound, Paclitaxel-hyaluronic Acid ... acid, taxane, ARC-100, Crolibulin, Comparidin disodium phosphate, Isofludelone, Flurapisin, MAP-4343, JJH-201601, larotaxel, ModraPac005 / r, Patupilone, xinalitaxel, azitaxel, ARC-01, BT-1769, CAP-6, and crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites of the foregoing drugs; Preferably, the tubulin inhibitor is selected from docetaxel, albumin-bound paclitaxel, colchicine, paclitaxel, vinblastine, albumin-bound docetaxel, paclitaxel poliglumex, paclitaxel-hyaluronic acid, and their crystalline forms, pharmaceutically acceptable salts, stereoisomers, tautomers, prodrugs, and metabolites; Preferably, the tubulin inhibitor is selected from albumin-bound paclitaxel, paclitaxel, Paclitaxel poliglumex, Paclitaxel-hyaluronic acid; Preferably, the tubulin inhibitor is selected from albumin-bound paclitaxel; Preferably, the Aurora B / VEGFR / PDGFR / c-Kit / CSF1R multi-target kinase inhibitor is selected from sioronib, or a crystalline form or a pharmaceutically acceptable salt thereof, and the unit dose of sioronib is about 1-100 mg, preferably about 5-80 mg, more preferably about 10-50 mg, and most preferably about 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg; calculated as sioronib; Preferably, the DNA blocking agent is selected from 5-fluorouracil, or a crystal form or a pharmaceutically acceptable salt thereof; the unit dose of the 5-fluorouracil is 10-500 mg; preferably 25-300 mg, and most preferably 50 mg, 125 mg or 250 mg, calculated as 5-fluorouracil; Preferably, the DNA blocking agent is selected from gemcitabine, or a crystal form or pharmaceutically acceptable salt thereof; the unit dose of gemcitabine is 100-2000 mg; preferably 200-1000 mg, and most preferably 200 mg or 1000 mg, calculated as gemcitabine; Preferably, the microtubule inhibitor is selected from paclitaxel or a crystal form or a pharmaceutically acceptable salt thereof; the unit dose of paclitaxel is 20 mg, 50 mg, 100 mg, 500 mg, 1 g, or 2 g, calculated in paclitaxel units; Preferably, the pancreatic cancer is locally advanced or metastatic pancreatic cancer; Preferably, the pancreatic cancer is pancreatic ductal adenocarcinoma.

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