阅读说明：本技术 一类具有抗肿瘤活性多肽及其制备方法与应用 (Polypeptide with anti-tumor activity, preparation method and application thereof ) 是由 胡宏岗 丛薇 汪楠 刘超 何世鹏 高飞 汤华 李英华 于 2020-06-23 设计创作，主要内容包括：本发明涉及医药技术领域,且公开了一类具有抗肿瘤活性多肽,具体指具有式(I)结构的多肽,具有式(I)结构的多肽及其药学上可接受的盐或酯：FLPILAX1LAAKFGPKLFX2LVX3KKX4 (I),其中,X1表示Ser或Ser(β-D-GlcNAc)；X2表示Cys或Pra或L-Aza或Asp或Dap或乙烯基甘氨酸；X3表示Thr或Thr(β-D-GlcNAc)；X4表示Cys或Pra或L-Aza或Asp或Dap或乙烯基甘氨酸；片段中Cys与Cys可形成或不形成二硫键环合；片段中Cys与乙烯基甘氨酸可形成或不形成硫醚键环合；片段中Pra与Aza可形成或不形成三氮唑环合；片段中Asp与Dap可形成或不形成酰胺键环合。该一类具有抗肿瘤活性多肽及其制备方法与应用,设计并合成了一系列新型Brevinin-1Bya环肽和糖肽类活性分子,旨在从多方面提高其理化性质以及抗肿瘤活性,提高肿瘤的治疗效果。(The invention relates to the technical field of medicines, and discloses a polypeptide with anti-tumor activity, in particular to a polypeptide with a structure shown in a formula (I), a polypeptide with a structure shown in a formula (I) and pharmaceutically acceptable salts or esters thereof: FLPILAX1LAAKFGPKLFX2LVX3KKX4 (I), wherein X1 represents Ser or Ser (. beta. -D-GlcNAc); x2 represents Cys or Pra or L-Aza or Asp or Dap or vinylglycine; x3 represents Thr or Thr (. beta. -D-GlcNAc); x4 represents Cys or Pra or L-Aza or Asp or Dap or vinylglycine; cys and Cys in the fragment may or may not form a disulfide bond ring; cys and vinyl glycine in the fragment may or may not form thioether bond cyclization; pra and Aza in the fragment may or may not form triazole cyclization; asp and Dap in the fragment may or may not form an amide bond ring. The polypeptide with anti-tumor activity, the preparation method and the application thereof design and synthesize a series of novel Brevinin-1Bya cyclopeptide and glycopeptide active molecules, and aim to improve the physicochemical property and the anti-tumor activity of the polypeptide and improve the treatment effect of tumors from multiple aspects.)

1. A polypeptide with anti-tumor activity is characterized in that the polypeptide has a structure shown in formula (I).

A polypeptide having the structure of formula (I) and pharmaceutically acceptable salts or esters thereof:

FLPILAX1LAAKFGPKLFX2LVX3KKX4 (I)

wherein X1 represents Ser or Ser (. beta. -D-GlcNAc); x2 represents Cys or Pra or L-Aza or Asp or Dap or vinylglycine; x3 represents Thr or Thr (. beta. -D-GlcNAc); x4 represents Cys or Pra or L-Aza or Asp or Dap or vinylglycine; cys and Cys in the fragment may or may not form a disulfide bond ring; cys and vinyl glycine in the fragment may or may not form thioether bond cyclization; pra and Aza in the fragment may or may not form triazole cyclization; asp and Dap in the fragment may or may not form an amide bond ring.

2. A pharmaceutical composition comprising a polypeptide having the structure of formula (I) as described above, wherein the pharmaceutical composition comprises a polypeptide according to claim 1.

3. The pharmaceutical composition containing the polypeptide having the structure of formula (I) as claimed in claim 2, further comprising a pharmaceutically acceptable diluent, excipient or carrier.

4. The pharmaceutical composition of claim 3, wherein the carrier is one or more of ethanol, glycerol or water.

5. The application of the polypeptide with anti-tumor activity utilizes the pharmaceutical composition of the polypeptide with the structure of formula (I) to prepare anti-tumor drugs.

Technical Field

The invention relates to the technical field of medicines, in particular to a polypeptide with anti-tumor activity, a preparation method and application thereof.

Background

Research reports that the skin of the amphibian contains a large number of polypeptide molecules with good biological activity and shows pharmacological activities such as antibacterial property, antivirus, antifungal property, antitumor property and the like, and the polypeptide compounds become research hotspots.

The antibacterial peptide Brevinin-1BYa is derived from skin secretion of Rana amurensis (Rana boylii), and researches report that Brevinin-1BYa has activity of resisting gram-negative and gram-positive bacteria and is considered as an antibacterial drug candidate with better prospect, the family of antibacterial peptides also show good anti-tumor cell growth activity, and are very suitable for further development and utilization as anti-tumor drugs, and documents report that a disulfide bond structure in Brevinin-1Bya plays an important role in the conformational stability of the polypeptide and enables the polypeptide to play the basis of pharmacological activity, however, the disulfide bond is easily influenced by the reducing environment (such as reducing glutathione and the like) in vivo, so that the conformational change of the polypeptide causes activity loss and the tumor treatment effect is influenced, and therefore, the polypeptide with anti-tumor activity, the preparation method and the application thereof are provided to solve the problems.

Disclosure of Invention

Technical problem to be solved

Aiming at the defects of the prior art, the invention provides a polypeptide with anti-tumor activity, a preparation method and application thereof, has the advantage of high anti-tumor activity, and solves the problems that disulfide bonds in the existing anti-tumor drugs are easily influenced by the reducing environment (such as reducing glutathione and the like) in vivo, so that the activity is lost due to the change of the conformation of the disulfide bonds, and the tumor treatment effect is influenced.

(II) technical scheme

In order to realize the aim of high antitumor activity, the invention provides the following technical scheme: a polypeptide with antineoplastic activity, in particular to a polypeptide with a structure shown in a formula (I).

A polypeptide having the structure of formula (I) and pharmaceutically acceptable salts or esters thereof:

FLPILAX1LAAKFGPKLFX2LVX3KKX4 (I)

wherein X1 represents Ser or Ser (. beta. -D-GlcNAc); x2 represents Cys or Pra or L-Aza or Asp or Dap or vinylglycine; x3 represents Thr or Thr (. beta. -D-GlcNAc); x4 represents Cys or Pra or L-Aza or Asp or Dap or vinylglycine; cys and Cys in the fragment may or may not form a disulfide bond ring; cys and vinyl glycine in the fragment may or may not form thioether bond cyclization; pra and Aza in the fragment may or may not form triazole cyclization; asp and Dap in the fragment may or may not form an amide bond ring.

The term "cyclic peptide and glycopeptide active molecule of the present invention" refers to a polypeptide of the present invention having the structure of formula (I), and such a polypeptide may be referred to as "cyclic peptide and glycopeptide active molecule", "polypeptide fragment" or "polypeptide of the present invention" herein.

The amino group at the N-terminus and the carboxyl group at the C-terminus and the amino acid side chain group of the polypeptide of formula (I) may be unmodified or modified without substantially affecting the activity of the polypeptide of the invention, such as forming a "pharmaceutically acceptable ester", the modification of the N-terminal amino group including, but not limited to, de-amino, N-lower alkyl, N-di-lower alkyl and N-acyl modifications, the modification of the C-terminal carboxyl group including, but not limited to, amide, lower alkyl amide, dialkyl amide and lower alkyl ester modifications, the amino group at the N-terminus of the polypeptide of the invention being acetylated, i.e., -Ac, and the carboxyl group at the C-terminus being amidated, i.e., -NH 2.

As used herein, the polypeptide and the amino acid and chemical group are commonly known in the art, wherein the abbreviations of the amino acids are as defined in Table 1, the structures of the specific amino acids are as defined in Table 2, and the amino acids are generally referred to as L-form amino acids unless otherwise specified.

TABLE 1 amino acid abbreviation table

TABLE 2 Special amino acid abbreviations Table

Amino acids	Abbreviations
		O- (2-acetylamino-2-deoxy- β -D-glucopyranose) -L-serine	Ser(β-D-GlcNAc)
O- (2-acetamido-2-deoxy- β -D-glucopyranose) -L-threonine	Thr(β-D-GlcNAc)
		L-propargylglycine	Pra
L-3-azido-alanine	Aza
		L-2, 3-diaminopropionic acid	Dap

"pharmaceutically acceptable salts" refers to salts of certain small acidic or basic compounds with polypeptides, which generally increase the solubility of the polypeptide, and which do not substantially alter the activity of the polypeptide, e.g., acids which typically form salts with the polypeptides of the invention are hydrochloric, phosphoric, sulfuric, acetic, succinic, maleic, and citric acids, and the like; bases capable of forming salts with the polypeptide of the present invention include hydroxides, ammonium and carbonates of alkali metals or alkaline earth metals, and the like.

The antitumor effect of the polypeptide of the present invention can be verified by conventional experimental methods in the field, such as cytological experiments, etc., in the specific embodiment of the present invention, preferably by cytological experiments such as CCK-8 method, through which the cyclic peptide active molecules of formula (I) related to the present invention are all found to have in vitro antitumor effect.

In addition, the invention also provides a pharmaceutical composition containing the polypeptide fragment with the structure of the formula (I), which can be used for antitumor therapy, may contain one or more, preferably only one, the composition may contain one or more pharmaceutically acceptable diluents, excipients or carriers, preferably the composition is in unit dosage form, such as tablets, films, pills, capsules (including sustained release or delayed release forms), powders, granules, syrups or emulsions, sterile injectable solutions, suspensions or lyophilized powders, aerosols or liquid sprays, drop delivery devices or suppositories, the active pharmaceutical ingredient may be combined with a non-toxic pharmaceutically acceptable inert carrier, such as ethanol, glycerol, water or a combination thereof, the cyclopeptide active molecule of formula (I) of the present invention is preferably used as a sterile aqueous solution for injection.

The pharmaceutical compositions of the present invention may be administered by any means known to those skilled in the art, such as oral, rectal, sublingual, pulmonary, transdermal, iontophoretic, vaginal and intranasal administration, and preferably the pharmaceutical compositions of the present invention are administered parenterally, such as subcutaneous, intramuscular or intravenous injection.

The names, structural formulae and mass spectral data of some of the preferred compounds synthesized in accordance with the present invention are shown in Table 3.

TABLE 3 name, structural formula and Mass Spectrometry data for Cyclic peptides and glycopeptide active molecules

For the purpose of facilitating understanding, the present invention will be described with reference to specific embodiments and drawings, it being expressly understood that the description is illustrative only and is not intended to limit the scope of the invention.

(III) advantageous effects

Compared with the prior art, the invention provides a polypeptide with anti-tumor activity, a preparation method and application thereof, and the polypeptide has the following beneficial effects:

the research reports that the glycosylation modification of the polypeptide can improve the hydrophilicity, the oral bioavailability, the enzyme tolerance and the conformation stability of the polypeptide, therefore, the glycosylation modification of the serine and the threonine sites in Brevinin-1Bya is carried out, and in conclusion, a series of novel Brevinin-1Bya cyclic peptides and glycopeptide active molecules are designed and synthesized, and the research aims to improve the physicochemical property and the antitumor activity of the polypeptide from multiple aspects and improve the treatment effect of tumors.

Drawings

FIG. 1 is a schematic diagram of the chemical structure of Brevinin-1BYa-1 in accordance with the present invention;

FIG. 2 is a schematic diagram of the chemical structure of Brevinin-1BYa-2 in accordance with the present invention;

FIG. 3 is a schematic diagram of the chemical structure of Brevinin-1BYa-3 in accordance with the present invention;

FIG. 4 is a schematic diagram of the chemical structure of Brevinin-1BYa-4 in accordance with the present invention;

FIG. 5 is a schematic diagram of the chemical structure of Brevinin-1BYa-5 in accordance with the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.