CN115677835A - Polypeptide with cancer growth inhibition activity, biological material and application thereof - Google Patents
Polypeptide with cancer growth inhibition activity, biological material and application thereof Download PDFInfo
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Abstract
The invention discloses a polypeptide capable of inhibiting cancer growth, a biological material and application thereof, and belongs to the technical field of biological medicines. The invention aims to solve the problem of regulating and inhibiting the growth of breast cancer cells, in particular to MDA-MB-231 cells of triple negative breast cancer. Specifically disclosed is a polypeptide, a pharmaceutically acceptable salt thereof, or a derivative thereof, wherein the peptide is any one of the following: a1 And the amino acid sequence is the polypeptide shown in the sequence 1; a2 A polypeptide having 80% or more identity to the polypeptide represented by A1) and having the ability to inhibit the growth of cancer cells, which is obtained by substituting and/or deleting and/or adding an amino acid residue in the peptide represented by A1); a3 A fusion polypeptide obtained by linking the N-terminus or/and the C-terminus of A1) or A2) to a protein tag. The breast cancer cells are cultured in the environment with the polypeptide, and the polypeptide can obviously inhibit the growth of the breast cancer cells.
Description
Technical Field
The application belongs to the technical field of biological medicines, and particularly relates to a polypeptide with cancer growth inhibition activity, a biological material and an application thereof.
Background
The biological peptide is a polypeptide which has small molecular weight, simple structure, is composed of a plurality of amino acids or dozens of amino acids and has certain biological activity. In the clinical tumor treatment process, biological polypeptide has the tumor inhibiting effect and is also called tumor inhibiting peptide. Tumor suppressor peptides are important nutrients in antitumor therapy at present and may also become a main research direction of future biological pharmacy. The source of biological drugs mainly comprises inhibitory peptide sources, and various inhibitory biological peptides are synthesized by plants, animals and biological engineering techniques depending on organisms. In the clinical application of the biological peptide, the biological peptide has better regulation and control effects on tumors, has specificity and broad antitumor effect, and can inhibit the growth and development of the tumors, specifically kill and inhibit tumor cells. The biological peptide has various effects and can be subjected to related artificial modification under the condition of special needs of organisms, so that the biological peptide has a better clinical application prospect.
Disclosure of Invention
The technical problem to be solved by the present application is how to inhibit the proliferation of cancer cells (e.g. triple negative breast cancer cells).
In order to solve the above problems, the present invention provides the following technical solutions:
the present application provides a polypeptide, a pharmaceutically acceptable salt thereof, or a derivative thereof, the polypeptide being any one of:
a1 Polypeptide with the amino acid sequence of sequence 1;
a2 A polypeptide having 80% or more identity to the polypeptide represented by A1) and having cancer cell growth inhibitory activity, which is obtained by substituting and/or deleting and/or adding an amino acid residue in the peptide of A1);
a3 A fusion polypeptide obtained by linking the N-terminus or/and the C-terminus of A1) or A2) to a protein tag.
In the above polypeptides, the protein-tag refers to a polypeptide or protein that is expressed by fusion with a target protein using in vitro recombinant DNA technology, so as to facilitate expression, detection, tracking and/or purification of the target protein. The protein tag may be a Flag tag, a His tag, an MBP tag, an HA tag, a myc tag, a GST tag, and/or a SUMO tag, among others.
In the above proteins, identity refers to the identity of amino acid sequences. The identity of the amino acid sequences can be determined using homology search sites on the Internet, such as the BLAST web pages of the NCBI home website. For example, in the advanced BLAST2.1, by using blastp as a program, setting the value of Expect to 10, setting all filters to OFF, using BLOSUM62 as a Matrix, setting Gap existence cost, per residual Gap cost, and Lambda ratio to 11,1 and 0.85 (default values), respectively, and performing a calculation by searching for the identity of a pair of amino acid sequences, a value (%) of the identity can be obtained.
In the above protein, the 80% or greater identity may be at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.
In the above protein, the sequence 2 (SEQ ID No. 2) is composed of 273 amino acid residues.
One or more amino acids of the polypeptide of the present invention may be substituted with amino acids having a D-form conformation, artificially modified amino acids, naturally occurring rare amino acids, etc., to improve the bioavailability, stability and/or cancer suppressive activity of the polypeptide. Wherein the D-form amino acid means an amino acid corresponding to the L-form amino acid constituting the protein; the artificially modified amino acid refers to common L-type amino acid which is modified by methylation, phosphorylation and the like and forms protein; the rare amino acids existing in nature include unusual amino acids constituting proteins and amino acids not constituting proteins, such as 5-hydroxylysine, methylhistidine, gamma-aminobutyric acid, homoserine and the like.
In the above polypeptide, a pharmaceutically acceptable salt or a derivative thereof, the polypeptide derivative is any one of:
c1 A linker obtained by connecting an amino terminal protecting group to the amino terminal of the polypeptide and/or connecting a carboxyl terminal protecting group to the carboxyl terminal of the polypeptide;
c2 A linker obtained by linking oligopeptide or lipophilic group or cholesterol to the carboxyl terminal of the polypeptide;
c3 A linker obtained by linking an oligopeptide or a lipophilic group or cholesterol to the amino terminus of the above-mentioned polypeptide;
c4 A linker obtained by linking an oligopeptide or a lipophilic group or cholesterol to both the amino terminus and the carboxyl terminus of the above-mentioned polypeptide.
In the above polypeptide, a pharmaceutically acceptable salt or a derivative thereof, the cancer cell is any one of:
h1 ), breast cancer cells;
h2 Triple negative breast cancer cells);
h3 MDA-MB-231 cells of triple negative breast cancer.
In order to solve the above problems, the present application also provides a biomaterial.
The biomaterial provided by the application is any one of the following:
b1 Nucleic acid molecules encoding the peptides;
b2 An expression cassette containing the nucleic acid molecule of B1);
b3 A recombinant vector containing the nucleic acid molecule of B1), or a recombinant vector containing the expression cassette of B2);
b4 A recombinant cell containing the nucleic acid molecule of B1), or a recombinant cell containing the expression cassette of B2), or a recombinant cell containing the recombinant vector of B3);
b5 A recombinant tissue containing the nucleic acid molecule of B1), or a recombinant tissue containing the expression cassette of B2), or a recombinant tissue containing the recombinant vector of B3);
b6 A recombinant organ containing the nucleic acid molecule of B1), or a recombinant organ containing the expression cassette of B2), or a recombinant organ containing the recombinant vector of B3);
b7 B1), or a recombinant individual containing the nucleic acid molecule according to B2), or a recombinant individual containing the expression cassette according to B2), or a recombinant individual containing the recombinant vector according to B3).
B8 A recombinant microorganism containing the nucleic acid molecule of B1), or a recombinant microorganism containing the expression cassette of B2), or a recombinant microorganism containing the recombinant vector of B3).
B1 In the nucleic acid molecule, the nucleotide sequence of the present invention encoding the above-described polypeptide can be easily mutated by a person of ordinary skill in the art using a known method, for example, directed evolution or point mutation. Those nucleotides which are artificially modified to have 80% or more than 80% identity to the nucleotide sequence of the polypeptide isolated according to the present invention are derived from the nucleotide sequence of the present invention and are equivalent to the sequence of the present invention as long as they encode the polypeptide and have the ability to inhibit the growth of cancer cells.
The 80% or more identity may be 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity.
Herein, identity refers to the identity of amino acid sequences or nucleotide sequences. The identity of the amino acid sequences can be determined using homology search sites on the Internet, such as the BLAST web pages of the NCBI home website. For example, in the advanced BLAST2.1, by using blastp as a program, the Expect value is set to 10, all filters are set to OFF, BLOSUM62 is used as a Matrix, the Gap existence cost, the Per residual Gap cost, and the Lambda ratio are set to 11,1 and 0.85 (default values), respectively, and a search is performed to calculate the identity (%) of amino acid sequences, and then the value (%) of identity can be obtained.
In the above biological material, the nucleic acid molecule of B1) may be a gene encoding the protein. B1 The coding sequence of the nucleic acid molecule, in particular the coding strand, is the DNA molecule shown in SEQ ID No. 1.
Herein, the vectors are well known to those skilled in the art, including but not limited to: plasmids, phages (e.g., lambda phage or M13 filamentous phage, etc.), cosmids (i.e., cosmids), ti plasmids, or viral vectors.
In the above-mentioned biological materials, the expression cassette described in B2) means DNA capable of expressing the gene in a host cell, and the DNA may include not only a promoter for initiating gene transcription but also a terminator for terminating gene transcription. Further, the expression cassette may also include an enhancer sequence.
In the above biological material, the recombinant cell of B4) may be a recombinant microorganism, a recombinant insect cell and/or a recombinant insect tissue and/or a recombinant insect organ and/or a recombinant insect individual.
B5 The recombinant tissue may be a recombinant insect tissue and/or a recombinant plant tissue and/or a recombinant animal tissue.
B6 The recombinant organ may be a recombinant insect organ and/or a recombinant plant organ and/or a recombinant animal organ.
B6 The recombinant individual can be a recombinant insect individual and/or a recombinant plant individual and/or a recombinant animal individual.
In order to solve the above problems, the present application also provides a composition.
The compositions provided herein comprise a polypeptide, derivative thereof, or pharmaceutically acceptable salt thereof, as described above, and/or a biomaterial as described above;
the composition has at least one of the following functions:
e1 Inhibit cancer cell proliferation;
e2 Preparing a product for inhibiting the proliferation of cancer cells;
e3 The medicine is used for preparing medicines for treating and/or preventing and/or assisting in treating cancers.
The composition of above, wherein the cancer cell or cancer is any one of:
h1 Breast cancer cells or breast cancer;
h2 Triple negative breast cancer cells or triple negative breast cancer cells;
h3 MDA-MB-231 cells of triple negative breast cancer.
As above, the composition further comprises a pharmaceutically acceptable carrier or adjuvant.
In practical applications, the polypeptide, its pharmaceutically acceptable salt, its derivative, polymer and composition of the present invention can be administered directly to patients as a medicine or mixed with a suitable carrier or excipient and then administered to patients for the purpose of inhibiting the growth of cancer cells. The carrier material herein includes, but is not limited to, water-soluble carrier materials (e.g., polyethylene glycol, polyvinylpyrrolidone, organic acids, etc.), poorly soluble carrier materials (e.g., ethyl cellulose, cholesterol stearate, etc.), enteric carrier materials (e.g., cellulose acetate phthalate, carboxymethyl cellulose, etc.). Among these, water-soluble carrier materials are preferred. The materials can be prepared into various dosage forms, including but not limited to tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, liposomes, transdermal agents, buccal tablets, suppositories, freeze-dried powder injections and the like. Can be common preparation, sustained release preparation, controlled release preparation and various microparticle drug delivery systems. In order to prepare the unit dosage form into tablets, various carriers well known in the art can be widely used. Examples of the carrier are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, kaolin, microcrystalline cellulose, aluminum silicate and the like; wetting agents and binders such as water, glycerin, polyethylene glycol, ethanol, propanol, starch slurry, dextrin, syrup, honey, glucose solution, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone and the like; disintegrating agents such as dried starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene, sorbitol fatty acid ester, sodium dodecylsulfate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibitors such as sucrose, glyceryl tristearate, cacao butter, hydrogenated oil and the like; absorption accelerators such as quaternary ammonium salts, sodium lauryl sulfate and the like; lubricants, for example, talc, silica, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, and the like. The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets. In order to prepare the dosage form for unit administration into a pill, various carriers well known in the art can be widely used. Examples of the carrier are, for example, diluents and absorbents such as glucose, lactose, starch, cacao butter, hydrogenated vegetable oil, polyvinylpyrrolidone, gelucire, kaolin, talc and the like; binders such as acacia, tragacanth, gelatin, ethanol, honey, liquid sugar, rice paste or batter, etc.; disintegrating agents, such as agar powder, dried starch, alginate, sodium dodecylsulfate, methyl cellulose, ethyl cellulose, etc. In order to prepare the unit dosage form into suppositories, various carriers known in the art can be widely used. As examples of the carrier, there may be mentioned, for example, polyethylene glycol, lecithin, cacao butter, higher alcohols, esters of higher alcohols, gelatin, semisynthetic glycerides and the like. For preparing the unit dosage form into preparations for injection such as solution, emulsion, lyophilized powder and suspension, all diluents commonly used in the art, for example, water, ethanol, polyethylene glycol, 1,3-propanediol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitol fatty acid ester, etc., can be used. In addition, for the preparation of isotonic injection, sodium chloride, glucose or glycerol may be added in an appropriate amount to the preparation for injection, and conventional cosolvents, buffers, pH adjusters and the like may also be added. In addition, colorants, preservatives, flavors, flavorings, sweeteners or other materials may also be added to the pharmaceutical preparation, if desired. The preparation can be used for injection administration, including subcutaneous injection, intravenous injection, intramuscular injection, intracavity injection and the like; for luminal administration, such as rectally and vaginally; administration to the respiratory tract, e.g., intranasally; administration to the mucosa. The above route of administration is preferably by injection.
The application of the polypeptide, the medicinal salt or the derivative thereof in preparing the medicine for inhibiting the proliferation of cancer cells.
The use as described above, wherein the cancer cell or cancer is any one of:
h1 Breast cancer cells or breast cancer;
h2 Triple negative breast cancer cells or triple negative breast cancer cells;
h3 MDA-MB-231 cells of triple negative breast cancer.
The application of the biological material in preparing the medicine for inhibiting the proliferation of cancer cells.
1 the use as described above, wherein the cancer cell or cancer is any one of:
h1 Breast cancer cells or breast cancer;
h2 Triple negative breast cancer cells or triple negative breast cancer cells;
h3 Triple negative breast cancer MDA-MB-231 cells.
As above, the inhibiting cancer cell growth may be logarithmic phase cancer cell growth. The environment for inhibiting the growth of cancer cells can be high glucose DMEM medium containing 10% fetal bovine serum at 37 deg.C with 5% CO 2 . The log phase cancer cells can be cultured in high glucose DMEM medium containing 10% fetal calf serum at 37 deg.C and 5% CO 2 And obtaining the product.
Advantageous effects
The research related to the antibacterial peptide is carried out in laboratories of the applicant for nearly ten years, the main focus of attention is the research on the inhibition effect and mechanism of LL-37 antibacterial peptide on clinical common tumors, the research on the inhibition effect and mechanism of LL-37 antibacterial peptide and cec-mag antibacterial peptide on staphylococcus aureus and pseudomonas aeruginosa planktonic bacteria and biofilm, and meanwhile, the subject group carries out amino acid optimization on the antibacterial peptide reported in some literatures on the basis of the research so as to find an antibacterial peptide sequence with better tumor inhibition activity. Artificially synthesizing a polypeptide sequence which is WKRWVRRWKRWLR-NH2 and is abbreviated as WR-13-1; it was found to be able to inhibit the growth of triple negative breast cancer MDA-MB-231 cells.
The cell culture with the triple negative breast cancer MDA-MB-231 added is carried out by taking a blank group without the triple negative breast cancer MDA-MB-231 added as a blank group, taking a group without the antibacterial peptide WR-13-1 added as a control group, taking a group with the antibacterial peptide WR-13-1 added in different concentrations as an experimental group and taking a group with the polypeptide GR-14 added in different concentrations as a control group. WR-13-1 is found to be capable of inhibiting the growth of MDA-MB-231 cells of triple negative breast cancer, and the inhibitory activity of the MDA-MB-231 cells is obviously weakened due to the change of the single amino acid.
Drawings
FIG. 1 shows the inhibitory effect of WR-13-1 (A) and GR-14 (B) on the proliferation of triple negative breast cancer cells MDA-MB 231; the concentration gradient of the bacterial peptide cell inhibition rate is an abscissa, and the cell inhibition rate is an ordinate.
Detailed Description
Related biological materials:
the WR-13-1 antibacterial peptide is a polypeptide with an amino acid sequence of WKRWVRRWKRWLR (sequence 1), is synthesized by Gill Biochemical (Shanghai) Limited according to the amino acid sequence, and the synthetic concentration is more than or equal to 95 percent.
The GR-14 antibacterial peptide has an amino acid sequence of GXKRIVQRIKIDXIR (X = biphenylalanine) (the polypeptide described in the sequence 2), is synthesized by Jier Biochemical (Shanghai) Limited company according to the amino acid sequence, and has the synthetic concentration of more than or equal to 95 percent.
The culture medium used in the DMEM experiments was high-glucose DMEM, the specification of which is given below.
High-glucose DMEM: purchase item number: SH30022.01B.
10% fetal bovine serum: purchase item number: ST30-3302p.
CCK-8 reagent: water-soluble tetrazolium salt, WST-8 (2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2,4-disulfophenyl) -2H-tetrazolium monosodium salt, purchased under trade designation TS545.
Triple negative breast cancer MDA-MB-231: purchased from the chinese academy of sciences cell bank, catalog No.: TCTU 227, deposited in the laboratory of clinical medicine institute, university of Ningxia medicine.
The related method comprises the following steps:
weighing and dissolving antibacterial peptide WR-13-1:
the molecular weight M of the antibacterial peptide WR-13-1 is 2011.43g/mol (the mass concentration of substances is C = n/V, and C = (M/M)/V) and the sequence is shown as the sequence 1.
Taking a high-pressure EP tube, weighing 1mg WR-13-1, and dissolving in 100 μ l ddH 2 In O water, (weighed according to the volume of WR-13-1 needed finally) WR-13-1 in the tube is the stock solution. Shaking for 30 seconds, centrifuging at low speed for 30 seconds to dissolve completely, and C 1 = (M/M)/V = (1 mg/2011.43 g/mol)/100ul =4972 μmol/l, after which it is diluted to the desired concentration. By C 1 V 1 ＝C 2 V 2 The dilution concentration is the working concentration required for the cells.
If 10ml of WR-13-1 with the final working concentration of 10umol/l is required to be prepared, 23 μ l of WR-13-1 stock solution is added into culture solution without serum or with low serum in a centrifuge tube to ensure that the final volume is 10ml. The tube was shaken for 30s,3000rpm, and centrifuged for 30s to dissolve it sufficiently.
The following examples were processed using SPSS11.5 statistical software and the results were expressed as mean ± standard deviation, with One-wayaanova test, with P < 0.05 (. Star) indicating a significant difference, P < 0.01 (. Star) indicating a very significant difference, and P <0.001 (. Star) indicating a very significant difference.
Example 1
Culturing of cancer cells
1. Culture of human breast cancer cell MDA-MB-231 cell
Inoculating the human breast cancer cell MDA-MB-231 cell into high-glucose DMEM medium containing 10% fetal calf serum, and making the cell have a content of 5% CO at 37 deg.C 2 To the logarithmic growth phase of the cells.
Detection of polypeptide inhibition of cancer cell growth
1. Cultivation of cells to be tested
Collecting the MDA-MB-231 cells in the logarithmic growth phase 24h in advance, re-suspending in high-glucose DMEM medium containing 10% fetal calf serum to prepare cell suspension, inoculating the cell suspension into a 96-well cell culture plate, and adding the cell suspension into each well by 100 mu l to make the cell number be 1 × 10 4 Per well. 37 ℃ C., 5% CO 2 And (3) continuously culturing the cells for 24 hours under the environment, observing the cells to uniformly grow in the holes under a microscope, observing about 80 percent of the cells in the holes to be uniformly distributed in the holes under the microscope, and carefully sucking excess supernatant by using a pipette to obtain the cell culture plate to be tested.
2. Detection of cancer cell growth
Taking a blank group without adding cells to be tested and antibacterial peptide WR-13-1, a control group without adding antibacterial peptide WR-13-1 and an experimental group with adding antibacterial peptide WR-13-1 with different concentrations, wherein each group comprises four duplicate wells. The above operations are repeated to perform cancer inhibition detection on the antibacterial peptide WR-13-1. The specific operation is as follows:
2.1 detection of antimicrobial peptide WR-13-1
Blank group: the blank plate was added to 200. Mu.l of high-glucose DMEM medium without WR-13-1 antimicrobial peptide and serum.
Experimental groups: the cell culture plate to be tested is taken, and 200 mul of high-glucose DMEM culture medium (solution prepared by taking the high-glucose DMEM culture medium without serum as a solvent and taking WR-13-1 antimicrobial peptide as a solute) containing no serum and containing the LWR-13-1 antimicrobial peptide at the concentrations of 5, 10, 15, 20, 25, 30, 35 and 40 mul mol/LWR-13-1 antimicrobial peptide is sequentially added into the experimental group.
Control group (also called control group): the cell culture plate to be tested is taken, and 200 mu l of serum-free high-glucose DMEM medium is added into a control group.
Each group of four complex holes
2.2 detection of antimicrobial peptide GR-14
Blank group: the blank plate was added to 200. Mu.l of high-glucose DMEM medium without GR-14 antimicrobial peptide and serum.
Experimental groups: the cell culture plate to be tested is taken, and 200 mu L of high-glucose DMEM culture medium (solution prepared by taking the high-glucose DMEM culture medium without serum as a solvent and taking GR-14 antibacterial peptide as a solute) containing no serum and containing the GR-14 antibacterial peptide at the concentrations of 5, 10, 15, 20, 25, 30, 35 and 40 mu mol/L is sequentially added into the experimental group.
Control group (also called control group): the cell culture plate to be tested is taken, and 200 mu l of serum-free high-glucose DMEM medium is added into a control group.
Each group of four multiple holes.
The plates obtained in 2.1 and 2.2 above were placed at 37 ℃ and 5% CO 2 The culture is continued for 24h under the environment.
And after 24h of culture, taking out the culture plate (96-well cell culture plate), sucking the redundant cell supernatant by a pipette, adding 100 mu l of serum-free high-glucose DMEM culture medium and 10 mu l of CCK-8 reagent into each well, incubating for 1h at 37 ℃, measuring the absorbance value at 450nm by using a microplate reader, and calculating the cell inhibition rate of the antibacterial peptide and the corresponding IC50 of the antibacterial peptide after the specific experimental data are attached.
Cell killing rate) = (control well absorbance value-experimental well absorbance value)/(control well absorbance value-blank well absorbance value) × 100%, the abscissa is the concentration gradient of the cell killing rate of the antibacterial peptide, the ordinate is the cell killing rate, and a killing rate curve is drawn. The results are shown in Table 1.
The results show that the antibacterial peptide WR-13-1 inhibits the proliferation of MDA-MB-231 cells of triple negative breast cancer (P < 0.001) in a concentration-dependent manner, and the IC50 is 8.886 mu M. Meanwhile, the other peptide GR-14 is another peptide in FK13 series peptides, and has no concentration-dependent inhibition effect on MDA-MB-231 cells with triple negative breast cancer (P > 0.05).
The following examples were processed using SPSS11.5 statistical software and the results were expressed as mean ± standard deviation, with One-wayaanova test, P < 0.05 (x) indicating a significant difference, P < 0.01 (x) indicating a very significant difference, and P <0.001 (x) indicating a very significant difference.
TABLE 1 inhibitory Effect of different concentrations of antibacterial peptides WR-13-1 and GR-14 on MDA-MB-231 cells (CCK-8 experiment)
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.
Claims (10)
1. A polypeptide, a pharmaceutically acceptable salt thereof, or a derivative thereof, wherein the polypeptide is any one of:
a1 Polypeptide of which the amino acid sequence is sequence 1;
a2 A polypeptide having 80% or more identity to the polypeptide represented by A1) and having an activity of inhibiting cancer cell proliferation, which is obtained by substituting and/or deleting and/or adding an amino acid residue in the peptide represented by A1);
a3 A fusion polypeptide obtained by linking the N-terminus or/and the C-terminus of A1) or A2) to a protein tag.
2. The polypeptide, pharmaceutically acceptable salt or derivative thereof according to claim 1, wherein: the polypeptide derivative of claim 1 which is any one of:
c1 A linker obtained by linking an amino-terminal protecting group to the amino terminus of the polypeptide of claim 1 and/or linking a carboxyl-terminal protecting group to the carboxyl terminus of the polypeptide of claim 1 or 2;
c2 A linker obtained by linking oligopeptide or lipophilic group or cholesterol to the carboxyl terminal of the polypeptide of claim 1;
c3 A linker obtained by linking oligopeptide or lipophilic group or cholesterol to the amino terminus of the polypeptide of claim 1;
c4 A linker obtained by linking an oligopeptide or a lipophilic group or cholesterol to both the amino terminus and the carboxy terminus of the polypeptide of claim 1.
3. The polypeptide, pharmaceutically acceptable salt or derivative thereof according to claim 1 or 2, wherein: the cancer cell is any one of the following:
h1 ), breast cancer cells;
h2 Triple negative breast cancer cells);
h3 MDA-MB-231 cells of triple negative breast cancer.
4. A biomaterial, characterized in that the biomaterial is any one of the following:
b1 Nucleic acid molecules encoding the peptide of claim 1;
b2 An expression cassette containing the nucleic acid molecule of B1);
b3 A recombinant vector containing the nucleic acid molecule of B1), or a recombinant vector containing the expression cassette of B2);
b4 A recombinant cell containing the nucleic acid molecule of B1), or a recombinant cell containing the expression cassette of B2), or a recombinant cell containing the recombinant vector of B3);
b5 A recombinant tissue containing the nucleic acid molecule of B1), or a recombinant tissue containing the expression cassette of B2), or a recombinant tissue containing the recombinant vector of B3);
b6 A recombinant organ containing the nucleic acid molecule of B1), or a recombinant organ containing the expression cassette of B2), or a recombinant organ containing the recombinant vector of B3);
b7 A recombinant individual containing the nucleic acid molecule according to B1), or a recombinant individual containing the expression cassette according to B2), or a recombinant individual containing the recombinant vector according to B3).
B8 A recombinant microorganism containing the nucleic acid molecule of B1), or a recombinant microorganism containing the expression cassette of B2), or a recombinant microorganism containing the recombinant vector of B3).
5. A composition comprising the polypeptide, derivative thereof, or pharmaceutically acceptable salt thereof of claim 1, and/or the biomaterial of claims 2-3;
the composition has at least one of the following functions:
e1 Inhibit cancer cell proliferation;
e2 Preparing a product for inhibiting the proliferation of cancer cells;
e3 The medicine is used for preparing medicines for treating and/or preventing and/or assisting in treating cancers.
6. The composition of claim 5, wherein the cancer cell or cancer is any one of:
h1 Breast cancer cells or breast cancer;
h2 Triple negative breast cancer cells or triple negative breast cancer cells;
h3 MDA-MB-231 cells of triple negative breast cancer.
7. Use of a polypeptide according to claim 1 or 2, a pharmaceutically acceptable salt thereof or a derivative thereof in the manufacture of a medicament for inhibiting the proliferation of cancer cells.
8. The use according to claim 7, wherein the cancer cell or cancer is any one of:
h1 Breast cancer cells or breast cancer;
h2 Triple negative breast cancer cells or triple negative breast cancer cells;
h3 MDA-MB-231 cells of triple negative breast cancer.
9. Use of the biomaterial of claim 4 in the manufacture of a medicament for inhibiting the proliferation of cancer cells.
10. The use according to claim 9, wherein the cancer cell or cancer is any one of:
h1 Breast cancer cells or breast cancer;
h2 Triple negative breast cancer cells or triple negative breast cancer cells;
h3 MDA-MB-231 cells of triple negative breast cancer.
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| CN117285604A (en) * | 2023-07-22 | 2023-12-26 | 湖南中晟全肽生化有限公司 | Polypeptide with inhibitory activity on triple negative breast cancer cells and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117285604A (en) * | 2023-07-22 | 2023-12-26 | 湖南中晟全肽生化有限公司 | Polypeptide with inhibitory activity on triple negative breast cancer cells and application thereof |
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