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CN-121991175-A - Human mucin 1 specific binding polypeptide and application thereof in preparation of antitumor targeted drugs

CN121991175ACN 121991175 ACN121991175 ACN 121991175ACN-121991175-A

Abstract

The invention provides a human mucin 1 specific binding polypeptide and application thereof in preparing an anti-tumor targeted drug, and belongs to the technical field of biological medicines. The amino acid sequence of the human mucin 1 specific binding polypeptide provided by the invention comprises an amino acid sequence shown as SEQ ID NO. 1, or a polypeptide with the same function, wherein one or more amino acids of the amino acid sequence shown as SEQ ID NO. 1 are substituted, deleted and/or added. The polypeptide has small molecular weight, is easy to modify and couple, is suitable for constructing a targeted drug carrying system, has low immunogenicity, stronger tissue penetrating capacity and more clinical application potential compared with an antibody, and provides a novel molecular tool for developing a human mucin 1 targeted disease (such as tumor) diagnostic reagent or therapeutic drug.

Inventors

  • ZHANG PING
  • QIN XIN
  • LIU QIANRONG
  • LIU MINGMING
  • ZHAO BAOMING
  • GONG WEI

Assignees

  • 西安培华学院
  • 湖北文理学院

Dates

Publication Date
20260508
Application Date
20260210

Claims (10)

  1. 1. A human mucin 1 specific binding polypeptide, wherein the amino acid sequence of the human mucin 1 specific binding polypeptide comprises the amino acid sequence shown in SEQ ID No.1, or an amino acid sequence with the same function obtained by substituting, deleting and/or adding one or more amino acids in the amino acid sequence shown in SEQ ID No. 1.
  2. 2. A method for preparing a human mucin 1 specific binding polypeptide according to claim 1, comprising the steps of biological Tao Xi, amplification of positive phage, DNA sequence determination and verification of the polypeptide encoding phage display.
  3. 3. The derivative of a human mucin 1-specific binding polypeptide according to claim 1, wherein the derivative is obtained by conventional modification of a side chain group, amino terminus or carboxyl terminus of the human mucin 1-specific binding polypeptide or by attachment of a tag to the human mucin 1-specific binding polypeptide.
  4. 4. A derivative of a human mucin 1-specific binding polypeptide according to claim 3, wherein the conventional modification comprises, but is not limited to, alkylation, amination, carboxylation, acylation, phosphorylation, sulfation, carbonylation, ubiquitination, fluorophore modification, biotin modification, polyethylene glycol modification.
  5. 5. A derivative of a human mucin 1-specific binding polypeptide according to claim 3, wherein the tag comprises, but is not limited to, a His6 tag, a FLAG tag, an HA tag, a Myc tag, a fluorescent protein tag.
  6. 6. The use of the human mucin 1-specific binding polypeptide of claim 1, the human mucin 1-specific binding polypeptide prepared by the method of claim 2, or the derivative of the human mucin 1-specific binding polypeptide of any one of claims 3 to 5 in the preparation of a medicament for treating a disease caused by overexpression of human mucin 1.
  7. 7. The use according to claim 6, wherein the diseases caused by overexpression of human mucin 1 include, but are not limited to, cancer, inflammation, fibrotic diseases and autoimmune diseases.
  8. 8. The use according to claim 7, wherein the cancer includes, but is not limited to, pancreatic cancer, breast cancer, lung cancer, colorectal cancer and ovarian cancer.
  9. 9. A medicament for treating diseases caused by overexpression of human mucin 1, which is characterized in that an active ingredient of the medicament comprises the human mucin 1 specific binding polypeptide of claim 1, the human mucin 1 specific binding polypeptide prepared by the preparation method of claim 2 or the derivative of the human mucin 1 specific binding polypeptide of any one of claims 3 to 5.
  10. 10. A detection reagent comprising the human mucin 1-specific binding polypeptide of claim 1, the human mucin 1-specific binding polypeptide prepared by the method of claim 2, or the derivative of the human mucin 1-specific binding polypeptide of any one of claims 3 to 5.

Description

Human mucin 1 specific binding polypeptide and application thereof in preparation of antitumor targeted drugs Technical Field The invention relates to the technical field of biological medicines, in particular to a human mucin 1 specific binding polypeptide and application thereof in preparing an anti-tumor targeted drug. Background The tumor targeting drug can enhance the enrichment of the drug at the focus part by specifically recognizing the tumor high-expression marker, thereby enhancing the curative effect and reducing the side effect. Mucin 1 (muc 1) is a transmembrane protein which is highly expressed in various epithelial tumors and has abnormal glycosylation, and has become an important molecular target for tumor targeted therapy. Currently, targeting molecules for MUC1 mainly include monoclonal antibodies and derivatives thereof. However, antibodies have large molecular weight, limited tissue penetration, high cost of preparation, and may cause immunogenic reactions, limiting clinical use. In contrast, the polypeptide molecules have the advantages of small molecular weight, easy synthesis and modification, low immunogenicity, good tissue penetrability and the like, are ideal targeting vectors, and the phage display peptide library technology can be used for screening small molecule polypeptides combined with specific targets, thereby providing an effective means for developing polypeptides targeting MUC 1. Disclosure of Invention Aiming at the defects of the prior art, the invention provides a specific binding polypeptide of human mucin 1 and application thereof in preparing antitumor targeted drugs. The invention takes MUC1 as a classical tumor marker as a target molecule, 1 polypeptide sequence is obtained by biopanning a phage display random 12 peptide library, and experiments such as phage ELISA and the like show that the polypeptide can specifically identify MUC1 and has good application prospect in the research and development of a disease treatment drug taking MUC1 as a target spot. In order to achieve the above purpose, the specific technical scheme of the invention is as follows: In a first aspect of the present invention, there is provided a human mucin 1 specific binding polypeptide, the amino acid sequence of which comprises the amino acid sequence shown in SEQ ID NO. 1, or an amino acid sequence having the same function obtained by substituting, deleting and/or adding one or more amino acids to the amino acid sequence shown in SEQ ID NO. 1. In a second aspect of the invention, there is provided a method for preparing said human mucin 1 specific binding polypeptide comprising the steps of biological Tao Xi, amplification of positive phage, determination of DNA sequence encoding phage-displayed polypeptide and verification. Further, the biopanning operation comprises the steps of adding a human mucin 1 solution into a 96-well plate, incubating at 4 ℃ overnight, adding a TBS solution containing BSA for the next day, sealing, washing for several times, adding a phage solution for incubation, washing and eluting, adding the eluted phage solution into an escherichia coli culture for amplification, purifying and determining the titer of amplified phage, and repeatedly performing a second round of screening and a third round of screening to obtain screened positive plaques. Further, the amplification of the positive phage is carried out by adding an overnight culture of Escherichia coli into a culture medium for dilution, dipping a plurality of well-separated blue plaques for inoculation culture, amplifying, and centrifuging to obtain the amplified positive phage. Further, the DNA sequence determination of the polypeptide presented by the encoding phage is carried out by extracting phage single-stranded DNA, sequencing by a sequencer by taking phage single-stranded DNA as a template to obtain phage single-stranded DNA sequence, and deducing the amino acid sequence of the polypeptide presented by the phage according to the phage single-stranded DNA sequence. Further, the verification is performed by ELISA. In a third aspect of the present invention, there is provided a derivative of the human mucin 1-specific binding polypeptide obtained by conventional modification of a side chain group, amino terminus or carboxyl terminus of the human mucin 1-specific binding polypeptide or by attachment of a tag to the human mucin 1-specific binding polypeptide. Further, the conventional modifications include, but are not limited to, alkylation, amination, carboxylation, acylation, phosphorylation, sulfation, carbonylation, ubiquitination, fluorophore modification, biotin modification, polyethylene glycol modification. Further, the tag includes, but is not limited to, a His6 tag, a FLAG tag, an HA tag, a Myc tag, a fluorescent protein tag. In a fourth aspect, the present invention provides the use of the human mucin 1 specific binding polypeptide or derivative thereof in the manufacture of a medicament for