Search

KR-102961363-B1 - Mesothelin-binding peptide and use thereof

KR102961363B1KR 102961363 B1KR102961363 B1KR 102961363B1KR-102961363-B1

Abstract

The present invention relates to a peptide that binds to mesocerin and its uses. The peptide of the present invention specifically binds to mesocerin and inhibits it, thereby exhibiting the effect of inhibiting the migration and metastasis of cancer cells. The peptide of the present invention was selected by using phage peptide display technology to identify a specific peptide that binds well to cells with high expression of human mesocerin, and it was confirmed that it inhibits the migration of cancer cells by binding to mesocerin. The peptide of the present invention is relatively stable in serum and shows high potential as an anticancer therapeutic agent, anticancer drug delivery probe, and cancer diagnostic probe that inhibits metastasis caused by the progression and migration of cancer in the future.

Inventors

  • 이병헌
  • 박민성

Assignees

  • 경북대학교 산학협력단

Dates

Publication Date
20260511
Application Date
20230201
Priority Date
20220210

Claims (20)

  1. A peptide that specifically binds to mesothelin, consisting of the amino acid sequence indicated by SEQ ID NO. 1.
  2. A polynucleotide encoding the peptide of claim 1.
  3. A recombinant vector comprising the polynucleotide of paragraph 2.
  4. Transformed organisms transformed with the recombinant vector of paragraph 3 (except humans).
  5. A composition for diagnosing pancreatic cancer comprising the peptide of claim 1 as an active ingredient.
  6. delete
  7. delete
  8. A pharmaceutical composition for inhibiting pancreatic cancer metastasis comprising the peptide of claim 1 as an active ingredient.
  9. A pharmaceutical composition according to claim 8, characterized in that the peptide specifically binds to mesocerin and is internalized into pancreatic cancer cells, thereby inhibiting the invasion and migration of pancreatic cancer cells.
  10. A drug delivery composition comprising the peptide of claim 1 as an active ingredient.
  11. A drug delivery composition according to claim 10, characterized in that the drug is a peptide drug or an anticancer agent.
  12. A drug delivery composition according to claim 11, characterized in that the peptide drug is a cytotoxic peptide having apoptosis or necrosis-inducing activity.
  13. delete
  14. A fusion peptide formed by combining the peptide of claim 1 and an apoptosis peptide consisting of the amino acid sequence indicated by SEQ ID NO. 2.
  15. A pharmaceutical composition for the prevention or treatment of pancreatic cancer comprising the fusion peptide of claim 14 as an active ingredient.
  16. A pharmaceutical composition for the prevention or treatment of pancreatic cancer comprising the fusion peptide of claim 14 and gemcitabine as active ingredients.
  17. delete
  18. delete
  19. delete
  20. A composition for imaging pancreatic cancer cells comprising the peptide of claim 1 as an active ingredient.

Description

Mesothelin-binding peptide and use thereof The present invention relates to a peptide that binds to mesocerin and uses thereof for the diagnosis and treatment of cancer and the inhibition of cancer metastasis. Pancreatic cancer is a leading cause of cancer-related deaths worldwide, and it is difficult to diagnose before extensive local invasion or metastasis to distant organs occurs. Furthermore, many patients diagnosed with pancreatic cancer present with symptoms not directly related to the cancer, such as loss of appetite, malabsorption, and vomiting, which complicates diagnosis and can lead to a worse prognosis. There is currently no precise tool to effectively diagnose pancreatic cancer, and due to its strong resistance to chemotherapy, the mortality rate is significantly higher than that of other cancers. Surgical resection is the primary treatment for pancreatic cancer, but the prognosis for patients after surgery is poor. Mesothelin is a protein with a molecular weight of approximately 40 kDa that is overexpressed in various solid tumors, particularly mesotheliomas, pancreatic cancer, ovarian cancer, lung cancer, and biliary tract cancer. In the pancreas, mesothelin was found primarily in pancreatic cancers, exhibiting tumor specificity by not being expressed in normal or chronic pancreatitis. Pancreatic cancer develops through various stages due to mutations or dysfunctions in numerous genes and proteins; mesothelin levels are primarily increased in advanced, or invasive, stages of pancreatic cancer. While mesothelin is thought to play a role in cell adhesion, its specific function remains largely unknown. Current knowledge suggests that mesothelin overexpression increases the invasion and migration of pancreatic cancer. The binding of MUC16 to mesothelin on the pancreatic cancer cell membrane activates MAPK p38 and increases MMP7 synthesis, thereby enhancing invasiveness and motility. In the absence of MUC16 expression in pancreatic cancer cell membranes, mesoserine induced the activation of the ERK pathway, thereby increasing MMP7 expression. Overexpression of mesoserine affects proliferation by increasing STAT3 activation and cyclin E expression. Furthermore, activation of AKT leads to NF-κB activation, which in turn increases IL-6 expression, affecting proliferation and survival; it also increases the levels of anti-apoptotic proteins Bcl-XL and Mcl-1 while decreasing the levels of apoptotic proteins BAX and BAD. Based on these findings, the functions of mesoserine appear to increase the proliferation, survival, and invasion of pancreatic cancer. Additionally, there is a significant association between the short survival rates of pancreatic cancer patients and the overexpression of mesoserine. Therefore, the detection and regulation of mesoserine would be useful for the diagnosis and treatment of pancreatic cancer, and the development of peptides that specifically bind to and regulate mesoserine would serve as a valuable tool for the diagnosis and treatment of pancreatic cancer. Figure 1 shows the expression of mesoserin in pancreatic cancer cell lines using a mesoserin antibody. (A) Results confirming mesoserin expression in pancreatic cancer cell lines AsPC-1, PANC-1, BxPC-3, and MiaPaCa-2 via immunofluorescence analysis. (B) Results confirming mesoserin expression in pancreatic cancer cell lines AsPC-1, PANC-1, BxPC-3, and MiaPaCa-2 via Western blot analysis. (C) Results confirming mesoserin expression in breast cancer cell lines MDA-MB231 and MCF7, cholangiocarcinoma cell line KKU-213, normal cell line HEK293T, and normal breast cell line MCF10A via immunofluorescence analysis. Figure 2 shows the results of peptide selection for mesoserin binding using phage display. (A) Results of confirming whether HEK293T cell lines transfected with DNA expressing mesoserin and untransfected cell lines express mesoserin through immunofluorescence analysis. (B) Results of performing phage display 5 times repeatedly using HEK293T cell lines transfected with DNA expressing mesoserin and untransfected cell lines. Figure 3 shows the results of phage ELISA analysis of 11 sequences extracted by phage display. (A) The results of organizing the 11 sequences extracted after repeating phage display 5 times into a table. (B) The results of measuring the absorbance of the 11 extracted sequences through phage ELISA analysis. Figure 4 shows the CTILWSLTC sequence, which specifically binds to mesoserin, synthesized into a peptide and named MSLN-pep. (A) The results of confirming TAMRA-labeled MSLN-pep in pancreatic cancer cell lines expressing mesoserin, such as AsPC-1, PANC-1, BxPC-3, and MiaPaCa-2, via immunofluorescence analysis. (B) The results of confirming MSLN-pep in HEK293T cell lines infected with DNA expressing mesoserin via immunofluorescence analysis. Figure 5 shows the results of mesocerin antibody and MSLN-pep being expressed in the same region. (A) The results of co-localization analysis showing whether me