US-20260125425-A1 - Carrier Peptide Fragment and Use Thereof

US20260125425A1US 20260125425 A1US20260125425 A1US 20260125425A1US-20260125425-A1

Abstract

The present disclosure provides a novel carrier peptide fragment with cell membrane permeability. The carrier peptide fragment disclosed herein includes an amino acid sequence: (SEQ ID No.: 1) RKLAVYWSTYKRSR.

Inventors

Nahoko BAILEYKOBAYASHI
Tetsuhiko Yoshida
Masahiro Iijima

Assignees

TOAGOSEI CO., LTD.

Dates

Publication Date: 20260507
Application Date: 20230905
Priority Date: 20220907

Claims (6)

1 . A carrier peptide fragment comprising the following amino acid sequence: (SEQ ID No.: 1) RKLAVYWSTYKRSR.
2 . A construct comprising: the carrier peptide fragment according to claim 1 ; and a foreign substance that is bonded to an N-terminus and/or a C-terminus of the carrier peptide fragment.
3 . The construct according to claim 2 , wherein the foreign substance is at least one kind of organic compound selected from the group consisting of a polypeptide, a nucleic acid, a dye, and a drug.
4 . The construct according to claim 2 , wherein the foreign substance is disposed on the C-terminus of the carrier peptide fragment.
5 . A method for transferring a target foreign substance from outside a eukaryotic cell into at least a cytoplasm of the cell, the method comprising the steps of: (1) preparing the construct according to claim 2 ; and (2) supplying the construct into a sample including a target eukaryotic cell.
6 . The method according to claim 5 , wherein the eukaryotic cell to which the construct is to be transferred is a human or nonhuman mammalian cell.

Description

CROSS-REFERENCE TO RELATED APPLICATION(S) This application is a national stage 371 application of PCT/JP2023/032320, filed on Sep. 5, 2023, which claims priority to and the benefit of Japanese Application Patent Serial No. 2022-141808, filed Sep. 7, 2022, the entire disclosures of which are hereby incorporated by reference. REFERENCE TO SEQUENCE LISTING SUBMITTED VIA EFS-WEB This application includes an electronically submitted sequence listing in XML format. The XML file contains a sequence listing entitled “TG23-015PCT” which was created on Jun. 13, 2023, and is 32.2 kilobytes in size. The sequence listing contained in this XML file is part of the specification and is hereby incorporated by reference herein in its entirety. TECHNICAL FIELD The present invention relates to a method for transferring (carrying) a foreign substance from outside a eukaryotic cell into the cell, and a carrier peptide fragment used in the method. BACKGROUND Conventionally, polypeptides and other foreign substances, particularly biologically active substances, have been transferred into the cells of humans and other mammals, etc., (eukaryotic cells) to change the characteristics of the cells (as well as the tissues and organs including the cells) or to improve and enhance the function of the cells. For example, WO2011/013699 discloses some peptides that can function as cell penetrating peptides (also referred to as “CPPs”) that can transfer a foreign substance into a cytoplasm through a cell membrane from outside a cell, which is a nucleolar localization signal (also referred to as “NoLS”) that has been known as a signal sequence that localizes protein into an intranuclear nucleolus. SUMMARY Incidentally, a construct including a CPP and a foreign substance may vary in cell penetrating efficiency depending on the combination between an amino acid sequence of the CPP and the kind of foreign substance. In recent years, techniques of drug delivery by CPPs have attracted attention and developments of various CPPs to enable selection of CPPs in accordance with foreign substances to be used (for example, oligonucleotide therapeutics and the like) have been desired. The art disclosed herein has been made in view of the above circumstances, and a main object of the present disclosure is to provide a novel carrier peptide fragment with cell membrane permeability. It is another object of the present disclosure to provide a construct for transferring a foreign substance including such a carrier peptide fragment. It is still another object of the present disclosure to provide a method for transferring a target foreign substance from outside a eukaryotic cell into at least a cytoplasm of the cell. A carrier peptide fragment disclosed herein is a carrier peptide fragment that is transferred from outside a eukaryotic cell into at least a cytoplasm of the cell. This carrier peptide fragment includes an amino acid sequence: RKLAVYWSTYKRSR (SEQ ID No.: 1). The carrier peptide fragment with such a structure is a synthetic peptide manufactured artificially that has been found out according to the present inventor's earnest examination. This carrier peptide fragment has the cell membrane permeability, and can transfer a foreign substance from outside a cell into the cell. In addition, in order to achieve the aforementioned object, the present disclosure provides a construct for transferring a foreign substance (hereinafter also referred to as “construct” simply) that is manufactured in order to transfer a target foreign substance from outside the eukaryotic cell into at least the cytoplasm of the cell. The construct disclosed herein includes the carrier peptide fragment disclosed herein and a foreign substance that is bonded to an N-terminus and/or a C-terminus of the carrier peptide fragment. With this construct, the target foreign substance can be transferred from outside the cell into the cell. In one aspect of the construct disclosed herein, the foreign substance can be at least one kind of organic compound selected from the group consisting of a polypeptide, a nucleic acid, a dye, and a drug. Here, the term “polypeptide” refers to a polymer with a structure in which a plurality of amino acids are linked by peptide bonds. The polypeptide is not limited by the number of peptide bonds (that is, the number of amino acid residues). In other words, the term “polypeptide” encompasses those generally called peptides with about 10 or more and less than 300 amino acid residues and those generally called proteins (typically, high molecular compounds with 300 or more amino acid residues). In this field, polypeptides and proteins are not distinguished strictly. In this specification, polymers formed of a plurality of amino acid residues (including oligomers) are collectively referred to as polypeptides. Moreover, the term “nucleic acid” refers to a nucleotide polymer and includes DNA and RNA. The term “nucleic acid” is not limited by the number of bases. In o