CN104177476B - The polypeptide of a kind of targeted human cancerous cell and application thereof - Google Patents
The polypeptide of a kind of targeted human cancerous cell and application thereof Download PDFInfo
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- CN104177476B CN104177476B CN201410437046.XA CN201410437046A CN104177476B CN 104177476 B CN104177476 B CN 104177476B CN 201410437046 A CN201410437046 A CN 201410437046A CN 104177476 B CN104177476 B CN 104177476B
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Abstract
The present invention relates to polypeptide and the application thereof of a kind of targeted human cancerous cell.Described polypeptide is as shown in below general formula: YX1X2X3X4X5C；The amino acid residue of described polypeptide is L-type and/or D type；The invention still further relates to the conjugate that obtained by described polypeptide and carrier conjugation and this polypeptide, its bivalent formed or multivalent for preparing treatment, prevent or diagnose the application in the medicine of cancer or imaging preparation.Many Toplink of the present invention and cancer cell marker people's Aminopeptidase N (Aminopeptidase N, APN) specific binding, compensate for the biological preparation such as antibody and prepare the shortcomings such as loaded down with trivial details, poor stability, somewhat expensive, penetration power are weak, can be used for preparing diagnostic reagent and the targeted probes of cancer.
Description
Technical field
The present invention relates to medicinal chemistry art, be specifically related to a peptide species and application thereof, particularly relate to a kind of target
To the polypeptide of human cancer cell and derived by this peptide and can product protein bound with people's aminopeptidase and above-mentioned many
Peptide or the application in preparing cancer therapy drug or imaging preparation of its derivative product.
Background technology
Cancer is one major causes of death in the whole world, and data show: the newly-increased about 14,100,000 example cancers in the whole world in 2012
Disease case, number of cancer deaths reaches 8,200,000, and by comparison, the data of 2008 are respectively 12,700,000 Hes
7600000.In world wide, the most common cancer of diagnosis is followed successively by pulmonary carcinoma (1,800,000,13%), breast carcinoma (170
Ten thousand, 11.9%) and colorectal cancer (1,400,000,9.7%), main lethal cancer be pulmonary carcinoma (1,600,000,19.4%),
Hepatocarcinoma (800,000,9.1%) and gastric cancer (700,000,8.8%).
The infiltration of cancerous cell and transfer are one of basic features of malignant tumor, are also clinical treatment failures and lead
Pathogenic people's main causes of death.The prerequisite of cancer metastasis is the formation of new vessels.It is therefore current,
The neoplasm targeted therapy generated for new vessels is the focus of current cancer research field.
The sugared egg that people's Aminopeptidase N (Aminopeptidase N, APN) is made up of 967 amino acid residues
In vain, APN kinds of tumor cells such as melanoma, ovarian cancer, carcinoma of prostate, colon cancer, cancer of pancreas,
The cell surface high level expression such as breast carcinoma, pulmonary carcinoma, invasion and attack, transfer and the tumor fresh blood to tumor cell
The formation of pipe plays an important role, and directly results in and participates in tumor cell invasion and shift with penetrating basement membrane
Process.
At present, the diagnosis of targeting and medicine for APN are mainly antibody drug.Although antibody is with it
Specific target tropism is extensively applied in clinical diagnosis, but also exists that to prepare loaded down with trivial details, less stable, expense high
The shortcomings such as expensive, penetration power is weak.Therefore, in order to improve the specificity to diagnosis of metastasis and treatment with accurate
Property, make up the defect of antibody, in the urgent need to seek for APN design Small-molecule probe, using as detection and
The effective ways for the treatment of cancer.
Polypeptide drug and diagnostic probe with low cost, molecular weight is little, good biocompatibility, penetrance are strong
Feature, shows the strongest superiority at aspects such as cancer target administration, cancer diagnosis, even shows and replace
Trend for antibody class diagnosis and treatment reagent.In cancer research, find the high specific polypeptides medicine for cancerous cell
Polypeptide probe affine with height has become key issue.Polypeptide by aminoacid as elementary cell, amino acid range
The multiformity of combination determines polypeptide in sequence, conformation, functionally vary.Therefore for neoplasm metastasis
Label APN, filters out the polypeptide with specific target tropism from high-throughout polypeptide libraries, then develops
Become diagnostic reagent and the medicine of cancer, be the effective way solving an above-mentioned difficult problem.
Summary of the invention
It is an object of the invention to provide a peptide species and application thereof, a kind of targeted human cancerous cell is many
Peptide and derived by this peptide and can product protein bound with people's aminopeptidase and aforementioned polypeptides or its derivative product
Product application in preparing cancer therapy drug or imaging preparation.
For reaching this goal of the invention, the present invention by the following technical solutions:
First aspect, the invention provides the polypeptide of a kind of targeted human cancerous cell, the aminoacid sequence of described polypeptide
Row formula is:
YX1X2X3X4X5C
Wherein, Y is tyrosine；X1It is hydrophilic amino acid, preferably valine or glutamic acid；X2It is parent
Aqueous aminoacid, preferably valine or glutamic acid；X3It is aromatic amino acid, preferably tyrosine；X4
It is basic amino acid, preferably histidine；X5It is neutral or hydrophilic amino acid, preferably leucine or silk
Propylhomoserin；C is cysteine.
Polypeptide of the present invention is from least 7 aminoacid of amino terminal by one of sequence 1-4 in sequence table
Residue forms；Preferably, described polypeptide is made up of the amino acid residue one of sequence 1-4 in sequence table Suo Shi.
The aminoacid sequence of four polypeptide of the present invention is respectively as follows: AP-1:YVEYHLC；AP-2:
YEKYHSC；AP-3:YVENGYC；AP-4:YEVGHRC.
The polypeptide of the present invention is possibly together with substituent group, and described substituent group is alkoxyl, alkanoyl or amide groups.
Preferably, in the present invention, the amino acid residue forming described polypeptide is L-type and/or D type.
Second aspect, present invention also offers a kind of polypeptide-conjugate, by many described in first aspect present invention
Peptide obtains with carrier conjugation；Described carrier is medicine, toxin, cytokine, radioelement, carrier
Albumen, enzyme, agglutinin, fluorophor, quantum dot or high specific absorbance chromophore.
Described carrier can select according to concrete purpose, and such as carrier can be medicine or cytokine
Deng, can be by medicine or cytokine target cancer cell, it is also possible to radioelement, carrier protein, enzyme, solidifying
Collection element, fluorophor, quantum dot live high specific absorbance chromophore as carrier.
Preferably, described carrier is enzyme or fluorophor；Described enzyme is preferably horseradish peroxidase；Described
Fluorophor is preferably Fluorescein isothiocyanate.
The polypeptide of the present invention is strong with cancer cell marker APN adhesion, and has good specificity.
The third aspect, present invention also offers a kind of test kit, and it comprises as described in the first aspect of the invention
Polypeptide-conjugate described in polypeptide or second aspect.
Test kit of the present invention can be used for detecting human cancer cell or people's Aminopeptidase N albumen.
Fourth aspect, present invention also offers bivalent that polypeptide as described in the first aspect of the invention formed or
Multivalent, it has the characteristic of targeted human Aminopeptidase N albumen.
Preferably, described bivalent or multivalent by being covalently or non-covalently connected formation with polymer.
Described polymer can select according to specific purposes, such as, can be Polyethylene Glycol (PEG) or ring
Dextrin etc..
5th aspect, present invention also offers described in polypeptide as described in the first aspect of the invention or fourth aspect
Bivalent or multivalent preparation for treat, prevent or diagnose cancer medicine or imaging preparation in answering
With.
Described cancer includes but not limited to: breast carcinoma, pulmonary carcinoma, gastric cancer, hepatocarcinoma or cervical cancer etc..
The polypeptide of the present invention has the effect of targeting APN albumen, can increase medicine as target head or be loaded with medicine
Content in APN positive cell such as the carrier of thing such as nano material, liposome etc., then add and pharmaceutically can connect
Novel more effective targeted anticancer medicine made by the adjuvant being subject to or adjuvant.
Compared with prior art, the present invention at least has the advantages that
APN positive cell is played targeting by many Toplink of the present invention, and selectivity is strong, and the present invention relates to
Polypeptide can use the method for chemosynthesis to prepare, and purity is high, and molecular weight is little, high specificity, without exempting from
Epidemic focus, safe and reliable.
Accompanying drawing explanation
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, AP-3 and AP-4
Combination to APN albumen respectively.
Fig. 3 be AP-1, AP-2, AP-3 and AP-4 respectively with APN positive cell SKOV-3 and APN
Negative cells 293A's is specific binding；
Wherein, Fig. 3-(a)～(d) be AP-1, AP-2, AP-3 and AP-4 respectively with APN positive cell SKOV-3
Specific binding, Fig. 3-(e) is that comparison polypeptide SP and APN positive cell SKOV-3 are without combining；
Fig. 3-(f)～(i) be AP-1, AP-2, AP-3 and AP-4 respectively with the spy of APN negative cells 293A
Anisogamy, Fig. 3-(j) is that comparison polypeptide SP and APN negative cells 293A is without combining.
Fig. 4 is the APN protein loci of AP-1, AP-2 specific recognition SKOV-3 cell surface.
Fig. 5 is that AP-1, AP-2, AP-3 and the AP-4 specificity respectively with human hepatoma cell line HepG2 is tied
Close.
Wherein: sequence 1-4 in AP-1, AP-2, AP-3 and AP-4 corresponding sequence table respectively.
Detailed description of the invention
Technical scheme is further illustrated below by detailed description of the invention.Those skilled in the art
It will be clearly understood that the only help of described embodiment understands the present invention, it is not construed as the concrete restriction to the present invention.
Experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
Building and the screening of APN target polypeptide of embodiment 1 polypeptide libraries
1. experimental apparatus and material
N-methylmorpholine (NMM), piperidines, trifluoroacetic acid (TFA), dichloromethane (DCM), 1,2,3-indantrione monohydrate,
Vitamin C, phenol, tetramethylurea hexafluorophosphate (HBTU), hexahydropyridine, tri isopropyl silane (TIS),
Dithioglycol (EDT), DMF (DMF), absolute ether, resin, methanol, various
Fmoc protected amino acid, Peptide systhesis pipe, shaking table, vacuum pump, Rotary Evaporators, mentioned reagent and material
Material obtains the most from commercial channels.
2. the synthesis of " pearl one thing " polypeptide libraries
Use Fmoc Solid-phase peptide synthesis synthesis polypeptide libraries, the peptide storehouse institute structure of screening APN target polypeptide
The chemical formula built is as shown in Figure 1.Concrete grammar be by the way of mixing is split point by aminoacid the most one by one
It is coupled on solid-phase resin, then under strong acid, side chain protecting group is removed, and then screen.
(1) Tentagel-S-NH of 200mg is weighed2Resin, circulates according to Solid phase peptide synthssis program, adds
The HBTU of HBTU, the Cys equivalent entering the Met equivalent of 200mg carries out reacting two circulations successively.Treat
After having reacted, resin is divided into 8 parts, to often pipe be separately added into the Asn of 40mg, Arg, Leu,
Asp, Gly, Ser, His, Tyr carry out coupling, after treating coupling, 8 pipe trees with the HBTU of equivalent
Fat mixes, deprotection, cleans.Again resin is divided into 8 parts, to often pipe be separately added into 40mg Asn,
Arg, Leu, Asp, Gly, Ser, His, Tyr carry out coupling with the HBTU of equivalent, after treating coupling,
8 pipe resin mixing, deprotection, clean.Again resin is divided into 8 parts, is separately added into 40mg to often pipe
The HBTU of Asn, Arg, Leu, Asp, Gly, Ser, His, Tyr and equivalent carry out coupling, treat idol
After connection, 8 pipe resin mixing, deprotection, clean.Again resin is divided into 4 parts, to often pipe point
Not adding the Val of 50mg, Glu, Ile, Lys carry out coupling with the HBTU of equivalent, after treating coupling,
4 pipe resin mixing, deprotection, clean.Again resin is divided into 4 parts, is separately added into 50mg to often pipe
Val, the HBTU of Glu, Ile, Lys and equivalent carry out coupling, after treating coupling, 4 pipe resins
Mixing, deprotection, cleans.Again resin is divided into 4 parts, to often pipe be separately added into the Phe of 50mg, Tyr,
Ala, Leu carry out coupling with the HBTU of equivalent, after treating coupling, 4 pipe resin mixing, deprotection.
In above-mentioned resin, it is sequentially added into dichloromethane, trifluoroacetic acid, deviates from side chain protecting group, added
The dry resin being loaded with peptide storehouse is standby.
(2) screening of the polypeptide specific binding with APN
With PBS, the peptide pearl in polypeptide libraries is cleaned 3 times；Skim milk with 5% on DL instrument to peptide pearl
(37 DEG C, 2h) are closed in mixing；With PBS peptide pearl 3 times；With being dissolved in 5% skim milk of PBS by 1:
The APND albumen of 1000 dilution Biotin labellings, then mix on DL instrument with peptide pearl and hatch (37 DEG C,
2h)；With PBS peptide pearl；Magnetic bead and positive peptide pearl with the streptavidin labelling of excess are on DL instrument
(37 DEG C, 1h) are hatched in mixing.With liquid-transfering gun, the peptide pearl bottom pipe is transferred in another pipe.After hatching
Peptide library is placed in 1.5mL centrifuge tube, and centrifuge tube is placed on magnetic frame.Positive peptide pearl suction affected by magnetic forces
Invest centrifuge tube sidewall, and negative polypeptide is owing to gravitational settling is at the bottom of EP pipe.
With liquid-transfering gun, the peptide pearl bottom EP pipe is transferred in another EP pipe.The peptide that will sub-elect through magnetic field
Pearl is chosen one by one, and single peptide pearl uses hydrogen bromide cracking, through MALDI-TOF-MS (ground substance assistant laser solution
Inhale ionization time of flight mass spectrometry) identify acquisition corresponding sequence information, it is thus achieved that and sequence includes AP-1:YVEYHLC；
AP-2:YEKYHSC；AP-3:YVENGYC；AP-4:YEVGHRC.Again close by above-mentioned sequence
Becoming, MALDI-TOF identifies and HPLC purification is for subsequent experimental.
Experimental example 1 is by surface plasma resonance imaging (SPRi) method detection polypeptide and people's Aminopeptidase N
(APN) affinity interaction of albumen
By the AP-1 of 1mg/mL, AP-2, AP-3, AP-4 and 1 × PBS (phosphate buffer) point to core
On sheet, overnight incubation under 4 DEG C of wet condition, then with 10 × PBS 10min, then with 1 × PBS
Clean 10min, finally clean 2 times with deionized water, each 10min, immerse 1 × PBS containing 5% milk
In, overnight incubation under the conditions of 4 DEG C, then with 10 × PBS 10min, 1 × PBS 10min,
Finally clean 2 times with deionized water, each 10min, dry up with nitrogen, machine (Plexera on cartridge chipHT surface plasma resonance imaging system).
Flowing passes sequentially through 1 × PBS, 2 × PBS, 1.25 μ g/mL, 2.5 μ g/mL, 5 μ g/mL, 10 μ g/mL mutually
With the APN purifying protein of 20 μ g/mL, record analysis SPRi signal.
Combination is as in figure 2 it is shown, AP-1, and AP-2, AP-3, AP-4 are along with the increasing of APN protein concentration
Adding, it gradually strengthens with the adhesion of APN albumen, and 1 × PBS increases the most therewith.This enforcement is described
Four polypeptide described in example have strong combination to APN, can be used for being correlated with as the polypeptide of targeting APN
Research application.
Experimental example 2AP-1, AP-2, AP-3, AP-4 and APN positive cell SKOV-3, APN are cloudy
The interaction of sexual cell 293A
Fluorescein isothiocyanate (FITC) conjugate of AP-1, AP-2, AP-3, AP-4 uses solid phase synthesis
Method obtains, the AP-1 obtained in embodiment 1, the many blaa polypeptides of AP-2, AP-3, AP-4 continues even
Connection episilon amino caproic acid.In the solution that pyridine/DMF/dichloromethane ratio is 1:5:7, will
FITC is with peptide pearl hybrid reaction overnight.
AP-1 is obtained, the FITC conjugate of AP-2, AP-3, AP-4 after trifluoroacetic acid cracks.Polypeptide
SP according to the method described above with FITC coupling, it is thus achieved that SP-FITC conjugate is as comparison.Use MALDI-TOF
Identify and HPLC purification is for subsequent experimental.
Abortion syndrome SKOV-3 is incubated at McCoy ' the s 5A training containing 100mL/L hyclone
Support in base, with 1 × 105Culture dish (Φ=35mm) at the bottom of the circular glass of cell concentration implantation of/mL, 37 DEG C,
5%CO2After cultivating 24h in cell culture incubator, discarding culture fluid, addition McCoy ' s 5A culture medium is molten
The FITC-AP-1 solved, FITC-AP-2, FITC-AP-3, FITC-AP-4 (1mg/mL, ice bath pre-cooling)；
Matched group adds the SP-FITC (ice bath pre-cooling) dissolved by McCoy ' s 5A culture medium of same molar ratio；
After ice bath lucifuge hatches 40min, discard polypeptide solution respectively, and wash 3 times with pre-cooling PBS.
Human embryonic kidney cell line's 293A cell is incubated in the DMEM culture medium containing 100mL/L hyclone,
With 1 × 105Culture dish (Φ=35mm) at the bottom of the circular glass of cell concentration implantation of/mL, 37 DEG C, 5%CO2Carefully
After born of the same parents' incubator cultivates 24h, discard culture fluid, add the FITC-AP-1 dissolved by DMEM culture medium,
FITC-AP-2, FITC-AP-3, FITC-AP-4 (1mg/mL, ice bath pre-cooling)；Matched group adds identical rubbing
The SP-FITC (ice bath pre-cooling) dissolved by DMEM culture medium of your concentration；Ice bath lucifuge hatches 40min
After, discard polypeptide solution respectively, and wash 3 times with pre-cooling PBS.
Glimmering with in laser scanning co-focusing microscope (Olympus FV1000-IX81, Japan) detection cell
Light is distributed.Result is as it is shown on figure 3, add AP-1, the SKOV-3 cell observation of AP-2, AP-3, AP-4
To green fluorescence, and compare the SKOV-3 cell of polypeptide SP and 263 cells and AP-1, AP-2, AP-3,
263 cells of AP-4 the most do not observe green florescent signal.
The above results illustrates, the FITC conjugate of AP-1, AP-2, AP-3, AP-4 and the knowledge of SKOV-3
Not being that sequence is narrow spectrum, the FITC conjugate of AP-1, AP-2, AP-3, AP-4 is with high expressed APN's
SKOV-3 cell is specific binding.Above four peptide species then do not have with the 293A cell of APN feminine gender
Specific binding.
The cell location of 2.AP-1, AP-2, AP-3, AP-4
In order to observe AP-1, AP-2, AP-3, AP-4 are in the location of SKOV-3 cell, with FITC-AP-1,
SKOV-3 cell has been carried out double by FITC-AP-2, FITC-AP-3, FITC-AP-4 and Hoechst33342
Dye experiment.Hoechst 33342 is the nuclear blue fluorescent dyes of a kind of display.
Result is as it is shown on figure 3, show AP-1, and AP-2, AP-3, AP-4 are combined in the thin of SKOV-3 cell
On after birth, this is identical with APN expressive site.
The specificity of experimental example 3AP-1, AP-2, AP-3, AP-4 and APN interacts
1. abortion syndrome SKOV-3 is incubated at McCoy ' the s 5A training containing 100mL/L hyclone
Support in base, with 1 × 105Culture dish (Φ=35mm) at the bottom of the circular glass of cell concentration implantation of/mL, 37 DEG C,
5%CO2Cell culture incubator is cultivated 24h.
2. 2h before transfection, changes McCoy ' the s 5A culture medium of serum-free into by cell culture medium.
3. 0.5 μ L lipofectamine 2000 is joined McCoy ' the s 5A culture medium of 100 μ L serum-frees
In, then the APNsiRNA of 1 μ g is joined in McCoy ' the s 5A culture medium of serum-free.Stand 5min,
Being slowly added into by siRNA solution in lipofectamine 2000 solution, mix gently, room temperature stands 15min.
4. mixed Transfection solution is added dropwise over culture dish at the bottom of glass (Φ=35mm), changes into after 4h containing 100
McCoy ' the s 5A culture medium of mL/L hyclone.
5., after cultivating 24h, discard culture fluid, the FITC-AP-1 that addition McCoy ' s 5A culture medium is dissolved,
FITC-AP-2 (1mg/mL), after ice bath lucifuge hatches 40min, discards polypeptide solution respectively, and uses pre-cooling
PBS washs 3 times.
Glimmering with in laser scanning co-focusing microscope (Olympus FV1000-IX81, Japan) detection cell
Light is distributed, and result is as shown in Figure 4.SKOV-3 cell positive with APN respectively for AP-1 and AP-2 has
Specific combination.Use RNA interference (RNAi) process after cell RNA i SKOV-3 the most not with
AP-1 or AP-2 is specific binding.Empirical tests, the SKOV-3 that AP-3 and AP-4 is the most positive with APN
Cell has specific binding.
The interaction of experimental example 4AP-1, AP-2, AP-3, AP-4 and APN positive cell HepG2
Human hepatoma cell line HepG2's cell is incubated in the DMEM culture medium containing 100mL/L hyclone,
With 1 × 105Culture dish (Φ=35mm) at the bottom of the circular glass of cell concentration implantation of/mL, 37 DEG C, 5%CO2
After cell culture incubator cultivates 24h, discard culture fluid, add the FITC-AP-1 dissolved by DMEM culture medium,
FITC-AP-2, FITC-AP-3, FITC-AP-4 (1mg/mL, ice bath pre-cooling)；Matched group adds identical rubbing
The SP-FITC (ice bath pre-cooling) dissolved by DMEM culture medium of your concentration；Ice bath lucifuge hatches 40min
After, discard polypeptide solution respectively, and wash 3 times with pre-cooling PBS.
Glimmering with in laser scanning co-focusing microscope (Olympus FV1000-IX81, Japan) detection cell
Light is distributed.
Result is as it is shown in figure 5, add AP-1, and the HepG2 cell observation of AP-2, AP-3, AP-4 is to green
Color fluorescence, the HepG2 cell of matched group does not the most observe green florescent signal, AP-1, AP-2 is described,
The FITC conjugate of AP-3, AP-4 and the identification of HepG2 are that sequence is narrow spectrum.Add AP-1, AP-2,
The 293A cell of AP-3, AP-4 does not observe green fluorescence, and AP-1, AP-2, AP-3, AP-4 are described
The HepG2 cell of FITC conjugate and high expressed APN be specific binding.
Can draw from experimental example 1-4, the polypeptide of the present invention has targeted human Aminopeptidase N protein positive tumor
The characteristic of cell, thus in actual applications, and can be able to kill using the polypeptide of the present invention as target polypeptide
The preparation hindering cancerous cell is puted together mutually or mixes, for the targeted therapy of tumor.
Applicant states, the present invention illustrates the process of the present invention by above-described embodiment, but the present invention
It is not limited to above-mentioned processing step, does not i.e. mean that the present invention has to rely on above-mentioned processing step and could implement.
Person of ordinary skill in the field is it will be clearly understood that any improvement in the present invention, to former selected by the present invention
The equivalence of material is replaced and the interpolation of auxiliary element, concrete way choice etc., all falls within the protection model of the present invention
Within the scope of enclosing and disclosing.
Claims (12)
1. the polypeptide of a targeted human cancerous cell, it is characterised in that described polypeptide is by the sequence in sequence table
Amino acid residue composition shown in one of 1-4.
Polypeptide the most according to claim 1, it is characterised in that form the amino acid residue of described polypeptide
For L-type and/or D type.
3. a polypeptide-conjugate, it is characterised in that described polypeptide-conjugate is by claim 1 or 2 institute
The conjugate that the polypeptide stated obtains with carrier conjugation；Described carrier is medicine, toxin, cytokine, radiation
Property element, carrier protein, enzyme, agglutinin, fluorophor, quantum dot or high specific absorbance chromophore.
Polypeptide-conjugate the most according to claim 3, it is characterised in that described carrier is enzyme or fluorescence
Group.
Polypeptide-conjugate the most according to claim 4, it is characterised in that described enzyme is Radix Cochleariae officinalis peroxidating
Thing enzyme；Described fluorophor is Fluorescein isothiocyanate.
6. a test kit, it is characterised in that described test kit comprises the polypeptide described in claim 1 or 2
Or the polypeptide-conjugate that one of claim 3-5 is described.
Test kit the most according to claim 6, it is characterised in that: described test kit is used for detecting people's cancer
Cell or people's Aminopeptidase N albumen.
The bivalent of polypeptide the most according to claim 1 and 2 formation or multivalent, it is characterised in that
Described bivalent or multivalent have the characteristic of targeted human Aminopeptidase N albumen.
Bivalent the most according to claim 8 or multivalent, it is characterised in that described bivalent or
Multivalent by being covalently or non-covalently connected formation with polymer.
Bivalent the most according to claim 9 or multivalent, it is characterised in that described polymer is
Polyethylene Glycol or cyclodextrin.
The bivalent that one of 11. polypeptide according to claim 1 and 2 or claim 8-10 are described
Or multivalent is used for diagnosing the application in the medicine of cancer or imaging preparation in preparation.
12. application according to claim 11, it is characterised in that described cancer is breast carcinoma, lung
Any one in cancer, gastric cancer, hepatocarcinoma or cervical cancer.
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| CN105198964B (en) * | 2015-10-12 | 2019-01-25 | 国家纳米科学中心 | A kind of cancer target polypeptide, preparation method and application |
| CN105330725B (en) * | 2015-11-06 | 2019-01-04 | 国家纳米科学中心 | A kind of pH response and polypeptide and its application for targeting human tumour vascular markers VEGFR2 |
| CN113512538B (en) * | 2021-04-21 | 2023-01-31 | 扬州大学 | Monoclonal antibody for resisting swine aminopeptidase N protein and application thereof |
Citations (2)
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| CN101781357A (en) * | 2009-01-21 | 2010-07-21 | 中国科学院化学研究所 | Polypeptide for detecting human cancer cells, and application thereof |
| CN101921313A (en) * | 2010-04-07 | 2010-12-22 | 武汉凯泰新生物技术有限公司 | Polypeptide for curing or preventing cancer or derivative product and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101781357A (en) * | 2009-01-21 | 2010-07-21 | 中国科学院化学研究所 | Polypeptide for detecting human cancer cells, and application thereof |
| CN101921313A (en) * | 2010-04-07 | 2010-12-22 | 武汉凯泰新生物技术有限公司 | Polypeptide for curing or preventing cancer or derivative product and application thereof |
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| Rapid Screening of Peptide Probes through In Situ Single-Bead Sequencing Microarray;Weizhi Wang et al;《Anal. Chem.》;20141105;第86卷;11854-11859 * |
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