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CN-122005811-A - Application of IGF2BP2-m6A-VCAN-TLR2 signal shaft in preparation of medicine for treating intrahepatic bile duct cancer lymphatic metastasis

CN122005811ACN 122005811 ACN122005811 ACN 122005811ACN-122005811-A

Abstract

The invention discloses application of an IGF2BP2-m6A-VCAN-TLR2 signal shaft in preparation of a medicament for treating intrahepatic duct cancer lymphatic metastasis. Aiming at the difficult problems of unknown lymphatic transfer mechanism and lack of effective targets of intrahepatic cholangiocarcinoma, the invention discovers and verifies a brand-new way for regulating and controlling expression of the multipotent proteoglycan from insulin-like growth factor 2 mRNA binding protein 2 through m6A modification, thereby activating Toll-like receptor 2 signals, promoting macrophages to secrete vascular endothelial growth factor C and driving lymphatic transfer. Based on this, the present invention provides inhibitors, particularly small molecule compounds 8010-8498, that target the signaling axis. Experiments show that the compound can obviously inhibit bile duct cancer cell migration, invasion and epithelial interstitial transformation, reduce macrophage secretion of vascular endothelial growth factor C, effectively inhibit tumor growth and lymphatic metastasis in a nude mouse popliteal fossa lymph node metastasis model and a spontaneous bile duct cancer model, and has a synergistic effect when being combined with gemcitabine and cisplatin.

Inventors

  • CHEN GANG
  • CHEN BO
  • WANG BOXIANG
  • Gan Yuqian
  • CHEN QIWEN
  • LI JIAJUN
  • WANG YIXUAN

Assignees

  • 温州医科大学附属第一医院

Dates

Publication Date
20260512
Application Date
20260415

Claims (10)

  1. Use of an IGF2BP2-m6A-VCAN-TLR2 signaling axis for the preparation of a medicament for treating intrahepatic cholangiocarcinoma lymphatic metastasis, wherein the medicament exerts a therapeutic or prophylactic effect by inhibiting the activity or expression of the IGF2BP2-m6A-VCAN-TLR2 signaling axis, wherein the IGF2BP2-m6A-VCAN-TLR2 signaling axis is modified from insulin-like growth factor 2 mRNA binding protein 2 to N6-methyladenosine mediated by the downstream target thereof, thereby inhibiting a biological functional pathway formed by binding of the pluripotent proteoglycan to Toll-like receptor 2, said inhibiting comprising inhibiting at least one of insulin-like growth factor 2 mRNA binding protein 2 function or expression, inhibiting the function or expression of the pluripotent proteoglycan, and inhibiting the function or expression of Toll-like receptor 2.
  2. 2. Use of the IGF2BP2-m6A-VCAN-TLR2 signaling shaft according to claim 1 for the preparation of a medicament for the treatment of lymphatic metastasis in intrahepatic bile duct cancer, wherein the medicament comprises an IGF2BP2 inhibitor, which IGF2BP2 inhibitor is a substance capable of specifically inhibiting the biological function or reducing the expression level of insulin-like growth factor 2 mRNA binding protein 2.
  3. 3. The use of IGF2BP2-m6A-VCAN-TLR2 signaling axis according to claim 2 for the preparation of a medicament for the treatment of intrahepatic cholangiocarcinoma lymphatic metastasis, wherein the IGF2BP2 inhibitor is a small molecule compound selected from the group consisting of compounds 8010-8498, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein the compounds 8010-8498 are obtained by multiple rounds of screening of a total of more than 162 ten thousand compounds comprising Chemdiv and TargetMol libraries of compounds using Schr dinger software, based on binding affinity scoring values, drug-like pentad, polar surface area, lipid-water partition coefficient, cardiotoxicity risk, drug metabolizing enzyme inhibition risk, and mode of interaction with insulin-like growth factor 2 mRNA binding protein 2 activity key amino acid residues Asn215, gly202 and Thr176 as targets.
  4. 4. The use of IGF2BP2-m6A-VCAN-TLR2 signaling shaft according to claim 3 for the preparation of a medicament for the treatment of intrahepatic bile duct cancer lymphatic metastasis, wherein the medicament further comprises at least one additional active ingredient selected from the group consisting of a chemotherapeutic agent comprising one or more of gemcitabine, cisplatin, oxaliplatin, 5-fluorouracil, capecitabine, irinotecan, docetaxel, paclitaxel, albumin-bound paclitaxel, tigogenin, an inhibitor of apoptosis ligand 1, cytotoxic T lymphocyte-associated protein 4, an immunotherapeutic agent comprising an inhibitor of fibroblast growth factor receptor, isocitrate dehydrogenase 1, isocitrate dehydrogenase 2, epidermal growth factor receptor, vascular endothelial growth factor receptor, MET proto-oncogene receptor tyrosine kinase.
  5. 5. The use of IGF2BP2-m6A-VCAN-TLR2 signaling axis according to claim 1 in the manufacture of a medicament for treating lymphatic metastasis in intrahepatic cholangiocarcinoma, wherein the medicament blocks chemotaxis and recruitment of tumor-associated macrophages by the concentration gradient of the pluripotent proteoglycan by inhibiting the binding of the pluripotent proteoglycan to the chemokine CCL 2.
  6. 6. The use of IGF2BP2-m6A-VCAN-TLR2 signaling shaft according to claim 1 in the manufacture of a medicament for the treatment of intrahepatic cholangiocarcinoma lymphatic metastasis, wherein the medicament blocks tumor-associated macrophage-related signalling thereby activated, reduces the secretion levels of tumor necrosis factor- α and interleukin-6, and thereby reduces the expression and secretion of vascular endothelial growth factor C induced by tumor necrosis factor- α and interleukin-6, ultimately inhibiting tumor-associated lymphangiogenesis.
  7. 7. The use of IGF2BP2-m6A-VCAN-TLR2 signaling axis according to claim 1 for the preparation of a medicament for the treatment of intrahepatic cholangiocarcinoma lymphatic metastasis, characterized in that the medicament down regulates the protein expression level of said pluripotent proteoglycans by inhibiting the stability regulation of the mRNA of said pluripotent proteoglycans by the N6-methyladenosine modification mediated by the insulin-like growth factor 2 mRNA binding protein 2, in particular the N6-methyladenosine modification site of said insulin-like growth factor 2 mRNA binding protein 2 recognizing and binding to a specific position of the 3 'untranslated region of said pluripotent proteoglycans mRNA, which binding is verified by MeRIP-seq binding transcriptome sequencing technology, RNA immunoprecipitation experiments and by a dual-luciferase reporter gene experiment employing a plasmid comprising the sequence of the mutation of the N6-methyladenosine modification site of the 3' untranslated region of wild-type or pluripotent proteoglycans mRNA.
  8. 8. The use of IGF2BP2-m6A-VCAN-TLR2 signaling shaft according to claim 1 for the preparation of a medicament for the treatment of intrahepatic bile duct cancer lymphatic metastasis, wherein the medicament further inhibits the epithelial-mesenchymal transition process of intrahepatic bile duct cancer cells, reducing the migration and invasiveness of tumor cells by inhibiting the phosphatidylinositol 3 kinase/protein kinase B signaling pathway activated by up-regulation of the insulin-like growth factor 2 mRNA binding protein 2 expression, the inhibition of the epithelial-mesenchymal transition process being manifested by up-regulation of the expression of the epithelial marker E-cadherin, and down-regulation of the expression of the mesenchymal marker N-cadherin and vimentin.
  9. 9. Use of IGF2BP2-m6A-VCAN-TLR2 signalling shaft according to any one of claims 1 to 8 for the preparation of a medicament for the treatment or prophylaxis of intrahepatic bile duct cancer lymphatic metastasis, wherein the medicament is administered by a method selected from intravenous injection, arterial infusion, oral administration, subcutaneous implantation, intratumoral injection or topical sustained release administration, and wherein the medicament is in a dosage form selected from tablets, capsules, granules, injections, liposomes, nanoparticles, microspheres or implants.
  10. 10. The use of IGF2BP2-m6A-VCAN-TLR2 signaling axis according to claim 9 for the preparation of a medicament for the treatment of intrahepatic cholangiocarcinoma lymphatic metastasis, characterized in that the use further comprises the use in the preparation of a medicament for the treatment or prevention of other malignancies accompanied by abnormal activation of IGF2BP2-m6A-VCAN-TLR2 signaling axis and a propensity to lymphatic metastasis, including but not limited to gastric cancer, colorectal cancer, pancreatic cancer, esophageal cancer, head and neck squamous cell carcinoma, lung cancer, breast cancer, ovarian cancer, prostate cancer, bladder cancer, melanoma, the effectiveness and mechanism of action of which is determined by verifying the inhibitory effect of the medicament on insulin-like growth factor 2 mRNA binding protein 2, multipotent proteoglycans, toll-like receptor 2 expression or activity in a cell line or animal model of such malignancies, and the effect on tumor cell proliferation, migration, invasion, epithelial-mesenchymal transition, macrophage recruitment, vascular growth factor C secretion and tube production.

Description

Application of IGF2BP2-m6A-VCAN-TLR2 signal shaft in preparation of medicine for treating intrahepatic bile duct cancer lymphatic metastasis Technical Field The invention relates to the technical field of tumor treatment, in particular to application of an IGF2BP2-m6A-VCAN-TLR2 signal shaft in preparation of a medicament for treating intrahepatic bile duct cancer lymphatic metastasis. Background Intrahepatic cholangiocarcinoma is a highly fatal primary liver malignancy originating from intrahepatic cholangiocarcinoma epithelial cells, deeply elucidates the molecular regulatory mechanism of intrahepatic cholangiocarcinoma lymph node metastasis, and develops a specific intervention strategy for this metastasis pathway, which has become an urgent clinical need for improving the overall prognosis of intrahepatic cholangiocarcinoma patients. N6-methyladenosine is the most abundant endogenous chemical modification mode of messenger RNA in eukaryotic cells, and the dynamic reversible modification process is synergistically regulated and controlled by a methyltransferase complex, demethylase and methylation recognition reading proteins. Insulin-like growth factor 2 mRNA binding protein 2 is an important N6-methyladenosine reading protein identified in recent years, and can enhance the stability of the target gene messenger RNA and promote the translation efficiency thereof by recognizing and binding to N6-methyladenosine modification sites in specific regions on the messenger RNA molecule. The existing research shows that the insulin-like growth factor 2 mRNA binding protein 2 is abnormally high expressed in various malignant tumors and is closely related to poor prognosis of patients, but the specific biological functions and molecular mechanisms of the insulin-like growth factor 2 mRNA binding protein 2 in the lymphatic metastasis process of intrahepatic cholangiocarcinoma have not been reported systematically so far. Multipotent proteoglycans are large chondroitin sulfate proteoglycans present in the extracellular matrix, as structural components of the extracellular matrix, which can be involved in regulating the recruitment and activation of immune cells in the tumor microenvironment. The document reports that the tumor cell-derived multipotent proteoglycan can bind to Toll-like receptor 2 on the surface of macrophages and activate downstream inflammatory signal paths, but whether the signal axes are involved in the functional remodeling and lymphangiogenesis of macrophages related to intrahepatic cholangiocarcinoma is not clearly studied at present. Tumor-associated lymphangiogenesis is a key initiation step in tumor lymphatic metastasis, vascular endothelial growth factor C is the most important lymphopoietic factor known to date, macrophages in the tumor microenvironment are the major cellular source of vascular endothelial growth factor C, however the upstream molecular events that regulate the phenotypic polarization of tumor-associated macrophages secretion of vascular endothelial growth factor C remain largely unexplained. At present, the research and development of drugs for N6-methyladenosine modified pathways are mainly focused on methyltransferase and demethylase inhibitors, and the research and development of small molecule targeted drugs for methylation reading proteins such as insulin-like growth factor 2 mRNA binding protein 2 are still in a starting stage. Clinical first-line chemotherapy of intrahepatic cholangiocarcinoma still takes gemcitabine combined with platinum drugs as the main treatment, which is difficult to effectively block the tumor lymphatic metastasis process and is often accompanied by acquired drug resistance. Therefore, there is a need to discover novel molecular targets capable of specifically interfering with lymphatic metastasis of intrahepatic cholangiocarcinoma and develop targeted drug candidates and combination therapeutic strategies with independent intellectual property rights. Disclosure of Invention In view of the above, the present invention provides the use of IGF2BP2-m6A-VCAN-TLR2 signaling axis in the preparation of a medicament for treating lymphatic metastasis of intrahepatic bile duct cancer, the medicament exerting a therapeutic or prophylactic effect by inhibiting the activity or expression of IGF2BP2-m6A-VCAN-TLR2 signaling axis, wherein the IGF2BP2-m6A-VCAN-TLR2 signaling axis refers to a biological functional pathway from insulin-like growth factor 2 mRNA binding protein 2-mediated modification of N6-methyladenosine to its downstream target, thereby constituting by binding of pluripotent proteoglycans to Toll-like receptor 2, the inhibition comprising at least one of inhibiting the function or expression of insulin-like growth factor 2 mRNA binding protein 2, inhibiting the function or expression of pluripotent proteoglycans, and inhibiting the function or expression of Toll-like receptor 2. Preferably, the medicament comprises an IGF2BP2 inhibitor, whi