CN104177476A - Polypeptide of target human cancer cells and application thereof - Google Patents
Polypeptide of target human cancer cells and application thereof Download PDFInfo
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- CN104177476A CN104177476A CN201410437046.XA CN201410437046A CN104177476A CN 104177476 A CN104177476 A CN 104177476A CN 201410437046 A CN201410437046 A CN 201410437046A CN 104177476 A CN104177476 A CN 104177476A
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Abstract
The invention relates to a polypeptide of target human cancer cells and an application thereof. The polypeptide is shown as the following general formula: YX1X2X3X4X5; and an amino acid residue of the polypeptide is L-shaped and/or D-shaped. The invention also relates to a coupling substance obtained by coupling the polypeptide with a carrier and an application of the polypeptide and a bivalent body and/or a multi-valance body formed by the polypeptide to preparation of drugs or imaging preparations for treating, preventing or diagnosing the cancer. The polypeptide can be specifically combined with a cancer cell marker human aminopeptidase N (Aminopeptidase N, APN), so that the defects of complexity in preparation, poor stability, high cost and poor penetrating power of biological agents such as antibodies are solved; and the polypeptide can used for preparing a diagnostic reagent and a target probe of the cancer.
Description
Technical field
The present invention relates to pharmaceutical chemistry field, be specifically related to a peptide species and application thereof, relate in particular to a kind of polypeptide of targeted human cancer cells and by this peptide derived and can be with the protein bound product of people's aminopeptidase and aforementioned polypeptides or its derivative product in the application of preparing in cancer therapy drug or video picture preparation.
Background technology
Cancer is the major causes of death in the whole world, data presentation: the newly-increased approximately 1,410 ten thousand routine cases of cancers in the whole world in 2012, and cancer mortality number reaches 8,200,000, and by comparison, the data of 2008 are respectively 1,270 ten thousand and 7,600,000.The common cancer of diagnosis is followed successively by lung cancer (1,800,000 in world wide, 13%), mammary cancer (1,700,000,11.9%) and colorectal cancer (1,400,000,9.7%), main lethal cancer is lung cancer (1,600,000,19.4%), liver cancer (800,000,9.1%) and cancer of the stomach (700,000,8.8%).
The infiltration of cancer cells and transfer are one of essential characteristics of malignant tumour, are also clinical treatment failure the major cause that causes patient death.The prerequisite of cancer metastasis is the formation of new vessel.Therefore current, the neoplasm targeted therapy generating for new vessel is the focus in current cancer research field.
People's Aminopeptidase N (Aminopeptidase N, APN) glycoprotein being formed by 967 amino-acid residues, APN in kinds of tumor cells as cell surface high level expressions such as melanoma, ovarian cancer, prostate cancer, colorectal carcinoma, carcinoma of the pancreas, mammary cancer, lung cancer, the formation of invasion and attack, transfer and tumour neovascularity to tumour cell plays an important role, and directly causes and participates in tumor cell invasion and penetrate basilar membrane the process shifting occurs.
At present, be mainly antibody drug for diagnosis and the medicine of the target of APN.Although antibody with its specific target tropism widespread use in clinical diagnosis, exists the shortcomings such as loaded down with trivial details, the less stable of preparation, somewhat expensive, penetration power be weak.Therefore, in order to improve specificity and the accuracy to diagnosis of metastasis and treatment, make up the defect of antibody, in the urgent need to seeking for APN design Small-molecule probe, using the effective ways as test-and-treat cancer.
The feature such as polypeptide drug and diagnostic probe are low with cost, molecular weight is little, good biocompatibility, penetrance are strong, shows very strong superiority at aspects such as cancer target administration, cancer diagnosis, has even shown the trend of alternative antibody class diagnosis and treatment reagent.In cancer research, find and become key issue for high specific polypeptides medicine and the high affine polypeptide probe of cancer cells.Polypeptide is by amino acid as elementary cell, and the diversity of amino acid permutation and combination has determined that polypeptide varies in sequence, conformation, function.Therefore for metastases marker APN, filtering out the polypeptide with specific target tropism from high-throughout polypeptide libraries, then develop into diagnostic reagent and the medicine of cancer, is the effective way that solves an above-mentioned difficult problem.
Summary of the invention
The object of the present invention is to provide a peptide species and application thereof, particularly a kind of polypeptide of targeted human cancer cells and by this peptide derived and can be with the protein bound product of people's aminopeptidase and aforementioned polypeptides or its derivative product in the application of preparing in cancer therapy drug or video picture preparation.
For reaching this goal of the invention, the present invention by the following technical solutions:
First aspect, the invention provides a kind of polypeptide of targeted human cancer cells, and the aminoacid sequence general formula of described polypeptide is:
YX
1X
2X
3X
4X
5C
Wherein, Y is tyrosine; X
1be hydrophilic amino acid, be preferably α-amino-isovaleric acid or L-glutamic acid; X
2be hydrophilic amino acid, be preferably α-amino-isovaleric acid or L-glutamic acid; X
3be die aromatischen Aminosaeuren, be preferably tyrosine; X
4be basic aminoacids, be preferably Histidine; X
5be neutrality or hydrophilic amino acid, be preferably leucine or Serine; C is halfcystine.
Polypeptide of the present invention is made up of from least 7 amino-acid residues of N-terminal one of sequence 1-4 in sequence table; Preferably, described polypeptide is made up of the amino-acid residue shown in one of sequence 1-4 in sequence table.
The aminoacid sequence of four polypeptide of the present invention is respectively: AP-1:YVEYHLC; AP-2:YEKYHSC; AP-3:YVENGYC; AP-4:YEVGHRC.
Polypeptide of the present invention also contains substituting group, and described substituting group is alkoxyl group, alkyloyl or amide group.
Preferably, in the present invention the amino-acid residue that, forms described polypeptide is L-type and/or D type.
Second aspect, the present invention also provides a peptide species conjugate, by the polypeptide described in first aspect present invention and carrier coupling and obtain; Described carrier is medicine, toxin, cytokine, radioelement, carrier proteins, enzyme, lectin, fluorophor, quantum dot or high specific absorbance chromophoric group.
Described carrier can be selected according to concrete object, such as carrier can be medicine or cytokine etc., can be by medicine or cytokine target cancer cell, also can use radioelement, carrier proteins, enzyme, lectin, fluorophor, quantum dot to live high specific absorbance chromophoric group as carrier.
Preferably, described carrier is enzyme or fluorophor; Described enzyme is preferably horseradish peroxidase; Described fluorophor is preferably fluorescein isothiocyanate.
Polypeptide of the present invention and cancer cells marker APN bonding force are strong, and have good specificity.
The third aspect, the present invention also provides a kind of test kit, and it comprises the polypeptide conjugate as described in polypeptide or the second aspect as described in first aspect present invention.
Test kit of the present invention can be used for detecting human cancer cell or people's Aminopeptidase N albumen.
Fourth aspect, bivalent or multivalent that the present invention also provides the polypeptide as described in first aspect present invention to form, it has the characteristic of targeted human Aminopeptidase N albumen.
Preferably, described bivalent or multivalent are by being covalently or non-covalently connected to form with polymer.
Described polymer can be selected according to specific purposes, for example, can be polyoxyethylene glycol (PEG) or cyclodextrin etc.
The 5th aspect, the present invention also provides bivalent or the application of multivalent in the medicine for the preparation for the treatment of, prevention or diagnosing cancer or video picture preparation as described in polypeptide or the fourth aspect as described in first aspect present invention.
Described cancer includes but not limited to: mammary cancer, lung cancer, cancer of the stomach, liver cancer or cervical cancer etc.
Polypeptide of the present invention has the effect of target APN albumen, can be used as target head increases medicine or the carrier that is loaded with medicine as the content in APN positive cell such as nano material, liposome, then adds pharmaceutically acceptable auxiliary material or adjuvant is made novel more effective targeted anticancer medicine.
Compared with prior art, the present invention at least has following beneficial effect:
Polypeptide of the present invention can play targeting to APN positive cell, and selectivity is strong, and the polypeptide the present invention relates to can adopt the method for chemosynthesis to prepare, and purity is high, and molecular weight is little, high specificity, and non-immunogenicity, safe and reliable.
Brief description of the drawings
Fig. 1 is that the peptide storehouse of screening APN target polypeptide builds chemical formula schematic diagram.
Fig. 2 is that surface plasma resonance imaging (SPRi) method detects AP-1, AP-2, and AP-3 and AP-4 be the keying action to APN albumen respectively.
Fig. 3 is AP-1, AP-2, AP-3 and AP-4 respectively with the specific binding of APN positive cell SKOV-3 and APN negative cells 293A;
Wherein, Fig. 3-(a)～(d) is AP-1, AP-2, AP-3 and AP-4 respectively with the specific binding of APN positive cell SKOV-3, Fig. 3-(e) is that contrast polypeptide SP and APN positive cell SKOV-3 are without combination;
Fig. 3-(f)～(i) is AP-1, AP-2, AP-3 and AP-4 respectively with the specific binding of APN negative cells 293A, Fig. 3-(j) is that contrast polypeptide SP and APN negative cells 293A are without combination.
Fig. 4 is AP-1, the APN protein loci of AP-2 specific recognition SKOV-3 cell surface.
Fig. 5 is AP-1, AP-2, AP-3 and AP-4 respectively with human hepatoma cell line HepG2's specific binding.
Wherein: AP-1, AP-2, the sequence 1-4 in AP-3 and the corresponding sequence table of AP-4 difference.
Embodiment
Further illustrate technical scheme of the present invention below by embodiment.Those skilled in the art should understand, described embodiment helps to understand the present invention, should not be considered as concrete restriction of the present invention.
The experimental technique using in following embodiment if no special instructions, is ordinary method.
Material, reagent etc. used in following embodiment, if no special instructions, all can obtain from commercial channels.
The structure of embodiment 1 polypeptide libraries and the screening of APN target polypeptide
1. laboratory apparatus and material
N-methylmorpholine (NMM), piperidines, trifluoroacetic acid (TFA); methylene dichloride (DCM), triketohydrindene hydrate, vitamins C; phenol; tetramethyl-urea hexafluorophosphate (HBTU), hexahydropyridine, tri isopropyl silane (TIS); dithioglycol (EDT); DMF (DMF), anhydrous diethyl ether; resin; methyl alcohol, various Fmoc protected amino acids, polypeptide composite tube; shaking table; vacuum pump, Rotary Evaporators, mentioned reagent and material all obtain from commercial channels.
2. " pearl one thing " polypeptide libraries is synthetic
Adopt the synthetic polypeptide libraries of Fmoc solid-phase peptide synthetic method, the constructed chemical formula in the peptide storehouse of screening APN target polypeptide as shown in Figure 1.Concrete grammar is that the mode of splitting point by mixing is coupled to amino acid on solid-phase resin randomly one by one, then under strong acid, side chain protected group is removed, and then is screened.
(1) take the Tentagel-S-NH of 200mg
2resin, according to the circulation of solid-phase polypeptide synthesis program, adds the HBTU of the Met equivalent of 200mg, and the HBTU of Cys equivalent reacts two circulations successively.After question response completes, resin is divided into 8 parts, adds respectively the HBTU of Asn, Arg, Leu, Asp, Gly, Ser, His, Tyr and the equivalent of 40mg to carry out coupling to every pipe, after treating coupling, 8 pipe resins are mixed, deprotection, cleans.Again resin is divided into 8 parts, adds respectively the HBTU of Asn, Arg, Leu, Asp, Gly, Ser, His, Tyr and the equivalent of 40mg to carry out coupling to every pipe, after treating coupling, 8 pipe resins are mixed, deprotection, cleans.Again resin is divided into 8 parts, adds respectively the HBTU of Asn, Arg, Leu, Asp, Gly, Ser, His, Tyr and the equivalent of 40mg to carry out coupling to every pipe, after treating coupling, 8 pipe resins are mixed, deprotection, cleans.Again resin is divided into 4 parts, adds respectively the Val of 50mg to every pipe, Glu, Ile, the HBTU of Lys and equivalent carries out coupling, after treating coupling, 4 pipe resins is mixed, and deprotection, cleans.Again resin is divided into 4 parts, adds respectively the Val of 50mg to every pipe, Glu, Ile, the HBTU of Lys and equivalent carries out coupling, after treating coupling, 4 pipe resins is mixed, and deprotection, cleans.Again resin is divided into 4 parts, adds respectively the HBTU of Phe, Tyr, Ala, Leu and the equivalent of 50mg to carry out coupling to every pipe, after treating coupling, 4 pipe resins are mixed to deprotection.
In above-mentioned resin, add successively methylene dichloride, trifluoroacetic acid, deviates from side chain protected group, and the dry resin that obtains being loaded with peptide storehouse is for subsequent use.
(2) with the screening of the polypeptide of APN specific binding
Peptide pearl in polypeptide libraries is cleaned 3 times with PBS; Skimmed milk with 5% mixes sealing (37 DEG C, 2h) to peptide pearl on DL instrument; Clean peptide pearl 3 times with PBS; , then mix on DL instrument with peptide pearl and hatch (37 DEG C, 2h) by the APND albumen of 1:1000 dilution Biotin mark with 5% skimmed milk that is dissolved in PBS; With PBS cleaning peptide pearl; On DL instrument, mix and hatch (37 DEG C, 1h) with positive peptide pearl with the magnetic bead of excessive streptavidin mark.Manage the peptide pearl of bottom transfers in another pipe with liquid-transfering gun handle.Peptide library after hatching is placed in 1.5mL centrifuge tube, and centrifuge tube is placed on magnetic frame.Positive peptide pearl is affected by magnetic force and is adsorbed in centrifuge tube sidewall, and negative polypeptide is because gravity settling is managed the end at EP.
Peptide pearl bottom EP being managed with liquid-transfering gun is transferred in another EP pipe.The peptide pearl sub-electing through magnetic field is chosen one by one, and single peptide pearl adopts hydrogen bromide cracking, obtains corresponding sequence information through MALDI-TOF-MS (substance assistant laser desorpted ionized flight time mass spectrum) qualification, obtains sequence and comprises AP-1:YVEYHLC; AP-2:YEKYHSC; AP-3:YVENGYC; AP-4:YEVGHRC.Again synthetic by above-mentioned sequence, MALDI-TOF qualification and HPLC purifying are for subsequent experimental.
Experimental example 1 detects the affinity interaction of polypeptide and people's Aminopeptidase N (APN) albumen by surface plasma resonance imaging (SPRi) method
By the AP-1 of 1mg/mL, AP-2, AP-3, AP-4 and 1 × PBS (phosphoric acid buffer) point is to chip, overnight incubation under 4 DEG C of wet condition, then uses 10 × PBS to clean 10min, then uses 1 × PBS to clean 10min, finally use washed with de-ionized water 2 times, each 10min, immerses containing in 1 × PBS of 5% milk overnight incubation under 4 DEG C of conditions, then use 10 × PBS to clean 10min, 1 × PBS cleans 10min, finally uses washed with de-ionized water 2 times, each 10min, dry up machine (Plexera on cartridge chip with nitrogen
hT surface plasma resonance imaging system).
Moving phase is successively by the APN purifying protein of 1 × PBS, 2 × PBS, 1.25 μ g/mL, 2.5 μ g/mL, 5 μ g/mL, 10 μ g/mL and 20 μ g/mL, record analysis SPRi signal.
Keying action as shown in Figure 2, AP-1, AP-2, AP-3, AP-4 is along with the increase of APN protein concentration, the bonding force of itself and APN albumen strengthens gradually, 1 × PBS does not increase thereupon.Illustrate that four polypeptide described in the present embodiment have strong combination to APN, can be used as the polypeptide of target APN for relevant research application.
Experimental example 2AP-1, AP-2, AP-3, the interaction of AP-4 and APN positive cell SKOV-3, APN negative cells 293A
AP-1, AP-2, AP-3, fluorescein isothiocyanate (FITC) conjugate of AP-4 adopts solid phase synthesis process to obtain, the AP-1 obtaining in embodiment 1, AP-2, AP-3, continues coupling epsilon-amino caproic acid on AP-4 polypeptide microballon.In the solution that is 1:5:7 in pyridine/DMF/methylene dichloride ratio, FITC and peptide pearl hybrid reaction are spent the night.
After trifluoroacetic acid cracking, obtain AP-1, AP-2, AP-3, the FITC conjugate of AP-4.Polypeptide SP according to the method described above with FITC coupling, obtain SP-FITC conjugate in contrast.Adopt MALDI-TOF qualification and HPLC purifying for subsequent experimental.
Be that SKOV-3 is incubated in McCoy ' the s 5A substratum containing 100mL/L foetal calf serum, with 1 × 10 by Proliferation of Human Ovarian Cell
5the cell concn of/mL is implanted culture dish (Φ=35mm) at the bottom of circular glass, 37 DEG C, 5%CO
2in cell culture incubator, cultivate after 24h, discard nutrient solution, add the FITC-AP-1 dissolving with McCoy ' s 5A substratum, FITC-AP-2, FITC-AP-3, FITC-AP-4 (1mg/mL, ice bath precooling); Control group adds the SP-FITC (ice bath precooling) dissolving with McCoy ' s 5A substratum of same molar ratio; Ice bath lucifuge is hatched after 40min, discards respectively polypeptide solution, and with precooling PBS washing 3 times.
By human embryonic kidney cell line 293A cell cultures in containing in the DMEM substratum of 100mL/L foetal calf serum, with 1 × 10
5the cell concn of/mL is implanted culture dish (Φ=35mm) at the bottom of circular glass, 37 DEG C, 5%CO
2in cell culture incubator, cultivate after 24h, discard nutrient solution, add the FITC-AP-1 dissolving with DMEM substratum, FITC-AP-2, FITC-AP-3, FITC-AP-4 (1mg/mL, ice bath precooling); Control group adds the SP-FITC (ice bath precooling) dissolving with DMEM substratum of same molar ratio; Ice bath lucifuge is hatched after 40min, discards respectively polypeptide solution, and with precooling PBS washing 3 times.
By the fluorescence distribution in laser scanning co-focusing microscope (Olympus FV1000-IX81, Japan) detection cell.Result as shown in Figure 3, adds AP-1, AP-2, and AP-3, the SKOV-3 cell observation of AP-4 is to green fluorescence, and SKOV-3 cell and 263 cells and the AP-1 of contrast polypeptide SP, AP-2, AP-3,263 cells of AP-4 all do not observe green fluorescence signal.
The above results explanation, AP-1, AP-2, AP-3, the FITC conjugate of AP-4 and the identification of SKOV-3 are that sequence is narrow spectrum, AP-1, AP-2, AP-3, the FITC conjugate of AP-4 and the SKOV-3 cell of high expression level APN are specific bindings.The 293A cell of above four peptide species and APN feminine gender does not have specific binding.
2.AP-1, AP-2, AP-3, the cellular localization of AP-4
In order to observe AP-1, AP-2, AP-3, AP-4 is in the location of SKOV-3 cell, and with FITC-AP-1, FITC-AP-2, FITC-AP-3, FITC-AP-4 and Hoechst33342 have carried out two experiments of dying to SKOV-3 cell.Hoechst 33342 is blue fluorescent dyes of a kind of showed cell core.
Result as shown in Figure 3, shows AP-1, AP-2, and AP-3, AP-4 is combined on the cytolemma of SKOV-3 cell, and this is identical with APN expressive site.
Experimental example 3AP-1, AP-2, AP-3, the specificity of AP-4 and APN interacts
1. Proliferation of Human Ovarian Cell is that SKOV-3 is incubated in McCoy ' the s 5A substratum containing 100mL/L foetal calf serum, with 1 × 10
5the cell concn of/mL is implanted culture dish (Φ=35mm) at the bottom of circular glass, 37 DEG C, 5%CO
2in cell culture incubator, cultivate 24h.
2. 2h before transfection, changes cell culture medium into McCoy ' the s 5A substratum of serum-free.
3. 0.5 μ L lipofectamine 2000 is joined in McCoy ' the s 5A substratum of 100 μ L serum-frees, then the APNsiRNA of 1 μ g is joined in McCoy ' the s 5A substratum of serum-free.Leave standstill 5min, siRNA solution is slowly joined in lipofectamine 2000 solution, mix gently, room temperature leaves standstill 15min.
4. mixed transfection solution is dropwise added to culture dish (Φ=35mm) at the bottom of glass, after 4h, change McCoy ' the s 5A substratum containing 100mL/L foetal calf serum into.
5. cultivate after 24h, discard nutrient solution, add the FITC-AP-1 dissolving with McCoy ' s 5A substratum, FITC-AP-2 (1mg/mL), ice bath lucifuge is hatched after 40min, discards respectively polypeptide solution, and washs 3 times with precooling PBS.
By the fluorescence distribution in laser scanning co-focusing microscope (Olympus FV1000-IX81, Japan) detection cell, result as shown in Figure 4.AP-1 and AP-2 have specific combination with the SKOV-3 cell of the APN positive respectively.Adopt RNA disturb (RNAi) cell RNA i after treatment SKOV-3 not with AP-1 or AP-2 specific binding.Empirical tests, AP-3 and AP-4 also have specific binding with the SKOV-3 cell of the APN positive.
Experimental example 4AP-1, AP-2, AP-3, the interaction of AP-4 and APN positive cell HepG2
Human hepatoma cell line HepG2's cell cultures is in containing the DMEM substratum of 100mL/L foetal calf serum, with 1 × 10
5the cell concn of/mL is implanted culture dish (Φ=35mm) at the bottom of circular glass, 37 DEG C, 5%CO
2in cell culture incubator, cultivate after 24h, discard nutrient solution, add the FITC-AP-1 dissolving with DMEM substratum, FITC-AP-2, FITC-AP-3, FITC-AP-4 (1mg/mL, ice bath precooling); Control group adds the SP-FITC (ice bath precooling) dissolving with DMEM substratum of same molar ratio; Ice bath lucifuge is hatched after 40min, discards respectively polypeptide solution, and with precooling PBS washing 3 times.
By the fluorescence distribution in laser scanning co-focusing microscope (Olympus FV1000-IX81, Japan) detection cell.
Result as shown in Figure 5, adds AP-1, AP-2, AP-3, the HepG2 cell observation of AP-4 is to green fluorescence, and the HepG2 cell of control group does not observe green fluorescence signal, and AP-1 is described, AP-2, AP-3, the FITC conjugate of AP-4 and the identification of HepG2 are that sequence is narrow spectrum.Add AP-1, AP-2, AP-3, the 293A cell of AP-4 does not observe green fluorescence, and AP-1 is described, AP-2, AP-3, the FITC conjugate of AP-4 and the HepG2 cell of high expression level APN are specific bindings.
Can draw from experimental example 1-4, polypeptide of the present invention has the characteristic of targeted human Aminopeptidase N protein positive tumour cell, thereby in actual applications, can be using polypeptide of the present invention as target polypeptide, put together mutually or mix with the preparation that can kill and wound cancer cells, for the targeted therapy of tumour.
Applicant's statement, the present invention illustrates processing method of the present invention by above-described embodiment, but the present invention is not limited to above-mentioned processing step, does not mean that the present invention must rely on above-mentioned processing step and could implement.Person of ordinary skill in the field should understand, any improvement in the present invention, and the selections of the equivalence replacement to the selected raw material of the present invention and the interpolation of ancillary component, concrete mode etc., within all dropping on protection scope of the present invention and open scope.
Claims (10)
1. a polypeptide for targeted human cancer cells, is characterized in that, the aminoacid sequence general formula of described polypeptide is:
YX
1X
2X
3X
4X
5C
Wherein, Y is tyrosine; X
1be hydrophilic amino acid, be preferably α-amino-isovaleric acid or L-glutamic acid; X
2be hydrophilic amino acid, be preferably α-amino-isovaleric acid or L-glutamic acid; X
3be die aromatischen Aminosaeuren, be preferably tyrosine; X
4be basic aminoacids, be preferably Histidine; X
5be neutrality or hydrophilic amino acid, be preferably leucine or Serine; C is halfcystine.
2. polypeptide according to claim 1, is characterized in that, described polypeptide is made up of the amino-acid residue shown in one of sequence 1-4 in sequence table.
3. polypeptide according to claim 1 and 2, is characterized in that, the amino-acid residue that forms described polypeptide is L-type and/or D type.
4. a peptide species conjugate, is characterized in that, described polypeptide conjugate is the conjugate being obtained by the polypeptide described in claim 1-3 any one and carrier coupling; Described carrier is medicine, toxin, cytokine, radioelement, carrier proteins, enzyme, lectin, fluorophor, quantum dot or high specific absorbance chromophoric group.
5. polypeptide conjugate according to claim 4, is characterized in that, described carrier is enzyme or fluorophor; Preferably, described enzyme is horseradish peroxidase; Described fluorophor is fluorescein isothiocyanate.
6. a test kit, is characterized in that, described test kit comprises polypeptide or the polypeptide conjugate described in claim 1-4 any one.
7. test kit according to claim 6, is characterized in that: described test kit is for detection of human cancer cell or people's Aminopeptidase N albumen.
8. the bivalent or the multivalent that form according to the polypeptide described in claim 1-3 any one, is characterized in that, described bivalent or multivalent have the characteristic of targeted human Aminopeptidase N albumen;
Preferably, described bivalent or multivalent are by being covalently or non-covalently connected to form with polymer;
Preferably, described polymer is polyoxyethylene glycol (PEG) or cyclodextrin.
9. the application in the medicine for the preparation for the treatment of, prevention or diagnosing cancer or video picture preparation according to polypeptide, bivalent or multivalent described in claim 1-3 or 8 any one.
10. application according to claim 9, is characterized in that, described cancer is any one in mammary cancer, lung cancer, cancer of the stomach, liver cancer or cervical cancer.
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| CN113512538A (en) * | 2021-04-21 | 2021-10-19 | 扬州大学 | Monoclonal antibody for resisting swine aminopeptidase N protein and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105198964A (en) * | 2015-10-12 | 2015-12-30 | 国家纳米科学中心 | Tumor targeted polypeptide, and preparation method and application thereof |
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| CN105330725A (en) * | 2015-11-06 | 2016-02-17 | 国家纳米科学中心 | Polypeptide with pH response and human tumor vessel marker VEGFR2 targeting functions and application of polypeptide |
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| CN113512538B (en) * | 2021-04-21 | 2023-01-31 | 扬州大学 | Monoclonal antibody for resisting swine aminopeptidase N protein and application thereof |
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| CN104177476B (en) | 2016-10-05 |
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