CN106831953A - A kind of polypeptide and its application - Google Patents
A kind of polypeptide and its application Download PDFInfo
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- CN106831953A CN106831953A CN201710044577.6A CN201710044577A CN106831953A CN 106831953 A CN106831953 A CN 106831953A CN 201710044577 A CN201710044577 A CN 201710044577A CN 106831953 A CN106831953 A CN 106831953A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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Abstract
The present invention relates to a kind of polypeptide, its preparation method and their application that can be used to treat prostatic disorders.
Description
It is related to sequence table
The application contains the sequence table of computer-reader form, and it passes through to carry to state and is incorporated herein.
Technical field
The present invention relates to it is a kind of can be used for treat prostatic disorders polypeptide, encode its polynucleotides and they
Using.
Technical background
Prostate cancer is one of the most common cancer of western countries male (Gronberg, Lancet 361:859-64
(2003)).Treatment on prostate cancer, can select the distinct methods such as operation, radiotherapy, endocrine therapy and chemotherapy.Above-mentioned treatment
Method improves survival rate unsatisfactory for improving patient's long term prognosis.Be transferred to for notice active high, no by many researchers
The research of good reaction antineoplastic polypeptide that is small, being not likely to produce drug resistance.But the stability of antineoplastic polypeptide, selectivity are limitations
Two principal elements of its application.Filtration, degradation of protease due to kidney etc. are acted on, and cause antineoplastic polypeptide
Stability is poor, half-life short, blood concentration are low, bioavilability is low.Simultaneously because the poor selectivity of antineoplastic polypeptide, can cause
The cell of human normal and tissue damage, there is certain toxic and side effect to human body.Therefore, work out effective method and improve polypeptide
The selectivity and stability of medicine are urgent problems in antineoplastic polypeptide medicament research and development, are had to its clinical practice important
Meaning.
Scientist is devoted to the research of target tumor peptide.Have now been found that various protease related to tumour in tumour
There is overexpression in cell or extracellular matrix, it has been found that organized enzyme have matrix metalloproteinase (MMP) and fibroblast
Activator protein (FAPot), also to tumor of prostate specificity and the antigen (PSA) or hK2 (human of expression high
Kallikrein 2), these tumor cell surface it is specific expressed or by tumor cell specific secrete small molecule antigens
The common feature of material is to be provided simultaneously with two kinds of characteristics of tumor-localizing and proteolytic enzyme.Generally, will have cytotoxic mother
Body medicine is condensed into the hydrolysis substrate of the fermentoid with polypeptide, medicine is preferably targeted to tumor tissues.
The content of the invention
The present invention relates to a kind of polypeptide, it is selected from the group：
A () includes SEQ ID NO:Amino acid sequence shown in 1 or by SEQ ID NO:Amino acid sequence composition shown in 1
Polypeptide；
B the amino acid sequence of () in (a) is derivative by (a) by replacing, lacking or add one or several amino acid
Polypeptide.
Polynucleotides the invention further relates to encoding such polypeptides, the carrier comprising the polynucleotides, host cell.
Treatment prostatic disorders spy is being prepared the invention further relates to pharmaceutical composition and aforementioned polypeptides comprising aforementioned polypeptides
It is not the application in the medicine for treat prostate cancer.
Brief description of the drawings
Fig. 1 is the ESI mass spectrograms of the LASAP-1 polypeptides prepared by synthesis in solid state of the invention.
Fig. 2 is the TEM figures of polypeptide LASAP-1 self assembly forms of the present invention.
The present invention relates to a kind of polypeptide, it is selected from the group：
A () includes SEQ ID NO:Amino acid sequence shown in 1 or by SEQ ID NO:Amino acid sequence composition shown in 1
Polypeptide；
B the amino acid sequence of () in (a) is derivative by (a) by replacing, lacking or add one or several amino acid
Polypeptide.
In other embodiment of the invention, the polypeptide that the present invention is provided is comprising the amino shown in SEQ ID NO.1
The polypeptide of acid sequence, and its modified polypeptide or its homeopeptide.
In other embodiments of the present invention, " SEQ ID NO are included：The polypeptide of the amino acid sequence shown in 1 " includes,
For example, by SEQ ID NO：The polypeptide of the amino acid sequence composition shown in 1, and by SEQ ID NO：Amino acid sequence shown in 1
The polypeptide that signal peptide sequence is constituted is added in row, and by SEQ IDNO：The N-terminal and/or C of the amino acid sequence shown in 1
Add the polypeptide that amino acid sequence obtained by appropriate flag sequence is constituted in end.
" modified polypeptide " in the present invention is referred to and is included in SEQ ID NO：Lacked in amino acid sequence shown in 1, taken
Generation, insert or add amino acid sequence obtained from one or several amino acid, and with treatment prostatic disorders activity egg
White matter.
In the preferred embodiment of the present invention, the modification to amino acid in described modified polypeptide or its homeopeptide
It is " conservative sex modification ".For example " conservative replaces " are referred under conditions of not material alterations protein active, by 1
Or other 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factors of more amino acid to be chemically similar.Can enumerate, certain hydrophobic residue is hydrophobic with other
Property residue substitution situation, by certain polar residues with identical charges other polar residues replace situation.
The functionally similar amino acid of this conservative replaces can be carried out, is all public in the corresponding technical field of each amino acid
Know.Specifically, as nonpolar (hydrophobicity) amino acid, can enumerate, alanine, valine, isoleucine, bright ammonia
Acid, proline, tryptophan, phenylalanine, methionine etc..As polarity (neutrality) amino acid, can enumerate, glycine, silk ammonia
Acid, threonine, tyrosine, glutamic acid, asparagine, cysteine etc..As the amino acid with positive charge (alkalescence), can be with
Enumerate, arginine, histidine, lysine etc..Additionally, as negative electrical charge (acidity) amino acid, can enumerate, aspartic acid, paddy
Propylhomoserin etc..
" homeopeptide " in the present invention is referred to and included and SEQ ID NO：1 amino acid sequence for representing has at least
85%, at least preferably 90%, at least preferably 92%, at least preferably 93%, at least preferably 94%, at least preferably 95%, it is at least excellent
96% is selected, at least preferably 97%, at least preferably 98%, at least preferably 99%, more preferably 100% homology (sequence identity)
Amino acid sequence.
For the present invention, the degree of sequence identity between two amino acid sequences is used such as EMBOSS software kits
(EMBOSS:The European Molecular Biology Open Software Suite, Rice etc., 2000, Trends
Genet.16:276-277), Needleman-Wunsch performed in the Needle programs of preferably 3.0.0 editions or more highest version
Algorithm (Needleman and Wunsch, 1970, J.Mol.Biol.48:443-453) determine.The optional parameters for using is breach
Open point penalty (gap open penalty) 10, gap extension penalty (gap extension penalty) 0.5 He
EBLOSUM62 (EMBOSS editions of BLOSUM62) substitution matrix." highest identity is labeled as using Needle
(longestidentity) output result (using-nobrief options to obtain) " is calculated such as homogeneity percentage
Under：
(same residue × 100)/(comparing the sum of breach in length-comparison)
Polypeptide of the present invention can be that natural, synthesis, semi-synthetic or restructuring is produced.Polypeptide of the invention can lead to
Cross genetic engineering, generated by known peptide symthesis or by the polypeptide with the appropriate peptidase digestion present invention.Preferably,
Polypeptide of the present invention can be routinely biological engineering method by host cell recombinant DNA sequence coding produce polypeptide product,
Can according to synthesis in solid state or liquid phase synthesis, for example can according to Steward and Young (Steward, J.M. and Young, J.D.,
Solid Phase Peptide Synthesis, 2nd Ed., Pierce Chemical Company, Rockford, I11.,
(1984)) method of description is closed with Applied Biosystem synthesizers or PioneerTM peptide synthesizers by Solid phase chemistry technology
Into.
Invention further provides the polynucleotides of encoding such polypeptides.The polynucleotides of the present invention can be used in vivo
Or the polypeptide of the external generation present invention as described above, or can be used to be attributable in the gene of the protein of the coding present invention
The gene therapy of the disease of genetic abnormality.Any form of the polynucleotides of the present invention all can be used, as long as it encodes the present invention
Polypeptide, including mRNA, RNA, cDNA, genomic DNA and chemical synthesis polynucleotides.The polynucleotides of the present invention include
DNA comprising designated nucleotide sequence and its cylinder and sequence, as long as the polypeptide of the gained DNA encoding present invention.
The polynucleotides of the present invention can by those skilled in the art will know that method prepare.For example, as many as present invention
Nucleotides can be prepared as follows：CDNA library is prepared from the cell of the polypeptide of the expression present invention, and with DNA (such as SEQ of the present invention
ID NO:1 or partial sequence 3) hybridized as probe.The preparation of cDNA library can be used, for example, with reference to Sambrook
Etc. what is write《Molecular cloning》, referring to the method described in CSH Press (1989)；Or, can use commercially available
CDNA library.CDNA library also can be prepared as follows：From the cell extraction RNA of the polypeptide of the expression present invention, according to the DNA of the present invention
Sequence (such as SEQ ID NO:3) 1 synthesize widow DNA, and performing PCR is entered by primer of the few DNA, and amplification coding is of the invention
Protein cDNA.
In addition, as the nucleotide sequencing of the cDNA to obtained by, can routinely determine the translated region encoded by the cDNA, and energy
It is readily derived the amino acid sequence of the polypeptide of the present invention.Further, using cDNA for obtaining or part thereof as probe, screening-gene
Group DNA library, just can isolated genomic DNA.
The present invention also provides a kind of engineering carrier containing the polynucleotides for encoding polypeptide of the present invention.The gene work
Cheng Zaiti can be general carrier or expression vector.Specifically, it is adaptable to the commercially available expression vector generally band of prokaryotic
Have and mark and cellular replication origin may be selected, with the promoters such as lacI, T7, λ PL and trp, and known cloning vector
Other genetic elements of pBR322 (ATCC37017).Such commercial vector includes pGEM (Promega) and pKK223-3
(Pharmacia).Can select to be derived from pBR322 according to selected appropriate promoter and structural gene sequence to be expressed
Suitable carrier.GST prokaryotic expression systems can also be used for the present invention.Carrier suitable for eukaryotic starts with eukaryotic
Son such as CMV, SV40, such carrier include that (horse big dragon, Di Chunhui, Pang Jian etc. (1991) high-tech is communicated pMT-hIL-3
11：26-29)、pQE-9(Qiagen)、pD10、pNH18A(Stratagene)、pKK233-3、pDR540、pRIT5
, and pcDNA3, pCI, pWLNEO, pSG (Stratagene), pSVL (Pharmacia) (Pharmacia).
The present invention also provides a kind of host cell containing carrier of the invention, and the host cell can be used for expressing this hair
Bright polypeptide, the host includes but is not limited to：Prokaryotic hosts, Escherichia coli, bacillus, streptomyces etc.；Eucaryon
Host, such as：Saccharomyces, aspergillus, insect cell such as fruit bat S2 and fall army worm Sf9, zooblast such as CHO, COS (monkey kidney
Fibroblast, Gluzman (Cell 23：175,1981) Human cell line such as PC-3 cell lines, DU145 cell lines and its
It can express the cell line of compatible vehicle.
The present invention also provides a kind of pharmaceutical composition, polypeptide, polynucleotides, load that it contains therapeutically effective amount of the present invention
Body, host cell；Pharmaceutically acceptable carrier or excipient if necessary.Pharmaceutically acceptable carrier or excipient refer to nothing
Malicious solid-state, semisolid or liquid filler, diluent, coating material or other pharmaceutical adjuncts.
In the preferred embodiment of the invention, for example, as needed, medicine can be oral, sugar coated tablet, glue Nang, the wine made of broomcorn millet are made
Agent and micro- glue Nang；Or parenteral, the form of the sterile vehicle or suspension of injection water or any other pharmaceutics acceptable liquid.
For example, compound can be mixed into the executory unit dosage form of acceptable medicine with the acceptable carrying agent of pharmaceutics or medium,
Specifically there are aqua sterilisa, physiological saline, vegetable oil, emulsifying agent, suspending agent, surfactant, stabilizer, aromatic, excipient, matchmaker
Jie's agent, preservative, binding agent etc..The quantity of active component is in the suitable dose in shown required scope in these preparations.
Can be made into the example of tablet or glue Nang additive therefors is, binding agent such as gelatin, cornstarch, bassora gum and Ah
Draw primary glue, excipient such as avicel cellulose, swelling agent such as cornstarch, gelatin and alginic acid；Lubricant such as stearic acid
Magnesium；Sweetener such as sucrose, lactose or saccharin；Aromatic such as peppermint.It is above-mentioned when unit dosage form is micro- glue Nang
Liquid phase carrying agent is may also include in composition, such as oil.It is all that Injectable sterile composition can follow conventional medicine execution catalytic agent
As distilled water for injection is prepared.Physiological saline, glucose and other isotonic liquids include adjuvant, and such as D- D-sorbites, D- are sweet
Dew sugar, PEARLITOL 25C and sodium chloride, can be used as aqueous solution for injection.These can be used in combination with appropriate solubilizer, such as
Alcohols, it is specific such as ethanol, polyalcohols such as propane diols and polyethylene glycol, nonionic surfactant such as polysorbate80
And HCO-50.Sesame oil or soybean oil can be used as oil-based liquid, and can be combined with the methyl benzoate as solubilizer, phenmethylol
Use, and can be with buffer solution such as phosphate buffer, sodium acetate buffer；Anodyne, such as procaine hydrochloride；Stabilization
Agent, such as phenmethylol；Prepared together with antioxidant.The parenteral solution of preparation can be fitted into suitable ampoule.
Can be used method well known to those skilled in the art that medicinal compound of the invention is applied into patient, such as artery
It is interior, intravenous, percutaneous injection, and intranasal, through bronchus, intramuscular or Orally administered.Dosage and application process are according to patient
Body weight and age and application process and change；And those skilled in the art can routinely be selected.If described chemical combination
The DNA can be inserted carrier for gene therapy and using the carrier to be treated by thing by DNA encoding.Dosage
It is different because of the body weight of patient, age and symptom with application process, but appropriate selection can be carried out by those skilled in the art.
Described herein and claimed invention is not limited to the scope of specific embodiment disclosed herein, these realities
The scheme of applying is intended only to illustrate several aspects of the invention.
Embodiment 1：The preparation of LASAP-1 polypeptides
The synthesis of LASAP-1 polypeptides of the present invention uses solid phase synthesis process, is 0.25 mM of polypeptide of synthesis, and its step is such as
Under：
(1) activated resin：The Wang resins of the amino acid of Fmoc protections will be connected (purchased from the limited public affairs in gill biochemistry Shanghai
Department) weigh up, pour into clean anhydrous solid phase reactor, 5ml DCM (dichloromethane) dissolving activation is added, overnight；
(2) resin is cleaned：The liquid in reactor is drained, 4ml DMF (DMF) fully shaking is added
1min, drains, and so operates 8 times repeatedly；The resin after a small amount of activation is collected to remain to do Kaiser detections；
(3) Fmoc protections are taken off：After draining solvent, the DMF solution for adding 4ml to contain 20% piperidines is placed in shaking table, shakes
Solvent is drained after 5min；DMF solutions of the 4ml containing 20% piperidines is added, shaking table is placed in, 20min is shaken；
(4) cleaning resin, removing piperidines：After draining solvent, 4ml DMF solutions are added, be placed in shaking table, shake 1min, after
Drain；So repeatedly, it is repeated 8 times until piperidines is completely removed；
(5) electronic balance weighs amino acid (amino acid of Fmoc protections) and coupling reagent to be accessed：By 4 times of ammonia of amount
Base acid, 3.9 times of HBTU (O- BTAs-tetramethylurea hexafluorophosphate), 4 times of HOBT (I-hydroxybenzotriazole) for measuring
It is dissolved in 4ml DMF, mixes to after being completely dissolved, be added in solid phase reactor and be sufficiently mixed with resin, shaking table vibrates five points
Clock；
(6) after then, it is 8 times of resin of DIEA (DIPEA) to add mole, after being sufficiently mixed, is put
In shaking table, clock reaction 2h, often connecing an amino acid need to repeat step 2-6；
(7) Kaiser detections, ninhydrin produces aubergine complex with ammonia or one-level amine, and Kaiser reagents include：6%
Ethanol solution of ninhydrin；80% phenol ethanol solution；The KCN pyridine solutions of 2%0.001M, reaction is completed in taking a small amount of (6)
When resin, and each 2-3 drops of three kinds of compositions in the resin in (2), plus Kaiser reagents, 100 DEG C of heating 1-2min, if presenting
Blue or bronzing show the amino for also dissociating, and conversely then represent that connection is complete；
(8) when peptide chain engagement is finished, after cleaning resin, with piperidines deprotection twice；
(9) resin is cleaned with DMF 10 times, each 4ml；Resin is cleaned with DCM again 10 times, each 4ml；
(10) sample after cleaning is dried up with nitrogen, is subsequently adding the 20%DMSO aqueous solution, is aoxidized 4 hours at room temperature；
(11) after the completion of question response, vacuum dried sample；
(12) after treating that sample drying is finished, transfer a resin into heart bottle, install magnetic stirring apparatus, heart bottle is solid
Set, be slowly added to the cutting reagent (trifluoroacetic acid for mixing：Ultra-pure water：Thioanisole：Phenol：Dithioglycol=82.5:5:
5:5:2.5), add magneton to be sufficiently stirred for, 12h is reacted at room temperature；
(13) question response is completed, and reactant is transferred in solid phase reactor, to react what is do not shifted in solid phase reactor
Resin, so stands 10min, and heart bottle is rinsed with TFA (trifluoroacetic acid), and all resins and solution are poured into solid phase reactor
In, after mixture is filtered under nitrogen flowing, filtrate is placed in round-bottomed flask, is dried up under nitrogen flowing；
(14) treat that the sample in round-bottomed flask is blown to sticky, remove nitrogen, about 20ml ice ether is poured into round-bottomed flask
Precipitated polypeptide, insoluble matter is fully broken up, then trim, is placed in refrigerated centrifuge, 8000rpm/min centrifugations at 4 DEG C
15min, abandons supernatant, is re-dissolved in being broken up in 20ml ice ether, centrifugation；Centrifugation 3 times is repeated operation, precipitation is vacuum dried,
Obtain polypeptide crude product；
(15) polypeptide crude product carries out purity analysis using analytic type HPLC, after purified with preparation HPLC；
(16) target polypeptides after purification are identified using ESI high resolution mass spectrums, its mass spectrogram as shown in figure 1, by
The molecular weight that figure measures synthesized LASAP-1 polypeptides is 3074.75, illustrates its amino acid sequence such as SEQ ID NO of the present invention:
Shown in 1.
Embodiment 2：The self assembly form of polypeptide LASAP-1
Form after transmission electron microscope observing polypeptide LASAP-1 self assemblies, its operating procedure is as follows：(1) compound concentration
It is 40 μM of LASAP-1 solution, stand overnight makes its self assembly at room temperature, prepares the phosphotungstic acid (PTA) of 20mg/mL, phosphorus tungsten
It is 6.0-7.0 that sour dye liquor need to adjust pH with 0.1M NaOH；
(2) the special copper mesh of TEM is taken out, polypeptide sample is dripped on copper mesh, water droplet is formed, after standing 1min, filter paper is used
Draw a small amount of liquid at copper mesh edge, when copper mesh into semi-moist state, a drop phosphotungstic acid dye liquor is then added dropwise again, dye 1.5min
Left and right, is blotted with filter paper, then copper mesh is positioned in culture dish and is dried；
(3) using the form after transmission electron microscope observing LASAP-1 self-assembling polypeptides, as shown in Fig. 2 containing the present invention
The solution of LASAP-1 polypeptides can be self-assembly of nanosized micelles in left at room temperature over night.
Referring to the amino acid sequence of LASAP-1 polypeptides of the present invention, can with the cysteine residues at position 9 at position 1
Disulfide bond is formed, and this forms self assembly polypeptide beneficial to polypeptide of the present invention, the polypeptide LASAP-1 with self assembly ability is in human blood
There is stability higher in clear.
Embodiment 3：The activity experiment of polypeptide LASAP-1
The cytoactive experiment of polypeptide is using mtt assay detection, and its experimental procedure is as follows：
(1) solvent is prepared：The preparation of MTT solution：5mgmL is prepared with PBS (hyclone, purchased from Gibco companies)-1's
MTT (tetrazolium bromide, purchased from Sigma companies), is wrapped with masking foil, and 30 minutes hydrotropies are stirred at room temperature, and is used after being completely dissolved
0.22 μM of membrane filtration, finally keeps in dark place at 4 DEG C；The preparation of three lysates：SDS (lauryl sodium sulfate) is
10g, the consumption of isobutanol 5mL, 10M hydrochloric acid are 0.12mL, are dissolved with distilled water and are made into 100mL solution；The preparation of culture medium：
90% DMEM culture mediums (being purchased from Gibco companies) and 10% FBS；
DMEM culture mediums be used for cultivate Hela cells (Human cervical cancer cell lines, purchased from Chinese Academy of Sciences's Shanghai cell bank) and
WPMY-1 cells (people's normal prostatic matrix immortalized cells, purchased from Chinese Academy of Sciences's Shanghai cell bank)；RPMI-1640 culture mediums
(being purchased from Gibco companies) is for cultivating LnCap cells (Human Prostate Cancer Cells, purchased from Chinese Academy of Sciences's Shanghai cell bank)；
(2) Hela cells in blake bottle, WPMY-1 cells and LnCap cells grow to more than 80%, add 1mL pancreatin
(being purchased from Sigma companies) digestion, is placed on 37 DEG C, and 5% CO2gas incubator culture, Microscopic observation cell dissociation is completed, plus
Enter after 4mL culture mediums terminate and be centrifuged, centrifugal rotational speed, time are respectively 900rpm/min, 4min；Supernatant is abandoned, culture medium is added
Gently blow and beat, cell is evenly dispersed in culture medium；
(3) number of cells is calculated with cell counter, and adds culture medium and adjust cell concentration to 5 × 104mL-1；
(4) obtained cell suspension bed board, it is 100 μ L that volume is added in each hole, per hole cell number 5000, is then placed
5%CO2, it is incubated 16-18 hours in 37 DEG C of incubators；
(5) it is incubated after terminating, the polypeptide drugs of various concentrations is prepared with serum free medium, 96 orifice plates is added, with cell
It is incubated 24 hours.Blank group only adds culture medium (acellular), and cellular control unit only adds culture medium, and experimental group adds different dense
The polypeptide drugs of degree；
(6) after reaction terminates, culture medium is suctioned out.The MTT that will be prepared before dilutes ten times with without phenol red full culture medium, so
100 μ L are added per hole afterwards, wherein blank group need not addition MTT.96 orifice plates are put into cell culture incubator；
(7) after reacting 4 hours, the three joint-trial agent (10%SDS+5% isobutanols+0.012mol/L of 100 μ L is added per hole
HCl), at room temperature, it is placed on 12-24 hours on shaking table and mixes, the OD values in 570nm is detected with ELIASA, with SPSS software meters
Calculate the IC50 values of various cells.
Toxicity of the LASAP-1 polypeptides of table 1 to each cell
| Cell | IC50(μM) |
| Hela (human cervical carcinoma cell) | 239.83±12.34 |
| LnCap (Human Prostate Cancer Cells) | 44.61±9.28 |
| WPMY-1 (normal human prostate cell) | 156.51±5.41 |
In above-mentioned three kinds of cells, human prostate cell WPMY-1 is normal cell, and hK2 (human are not secreted
Kallikrein 2) enzyme；LnCap is Human Prostate Cancer Cells, secretes hK2 enzymes；Hela cells are human cervical carcinoma cell, are not secreted
HK2 enzymes.
As shown in table 1, LASAP-1 polypeptides of the present invention are minimum to secreting the IC50 values of the LnCap cells of hK-2 enzymes.
SEQUENCE LISTING
<110>Beijing University of Chemical Technology
<120>A kind of polypeptide and its application
<130>
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 28
<212> PRT
<213>Artificial sequence
<400> 1
Cys Arg Gly Asp Lys Gly Pro Asp Cys Gly Lys Ala Phe Arg Arg Phe
1 5 10 15
Leu Gly Ala Leu Phe Lys Ala Leu Ser His Leu Leu
20 25
Claims (10)
1. the polypeptide being selected from the group：
A () includes SEQ ID NO:Amino acid sequence shown in 1 or by SEQ ID NO:It is many that amino acid sequence shown in 1 is constituted
Peptide；
B the amino acid sequence of () in (a) is by replacing, lacking or add one or several amino acid as polypeptide derived from (a).
2. polypeptide described in claim 1, it is by SEQ ID NO:Amino acid sequence composition shown in 1.
3. the polynucleotides of polypeptide described in claim 1 or 2 are encoded.
4. the carrying agent of polynucleotides described in claim 2 is included.
5. comprising the host cell of carrier described in polynucleotides described in claim 2 or claim 3.
6. the preparation method of polypeptide described in claim 1 or 2, its with synthesis in solid state resin as initiation material, by Fmocization
Learn synthetically prepared.
7. pharmaceutical composition, polypeptide is used as active component and pharmaceutics described in its claim 1 or 2 for including pharmaceutical effective amount
Acceptable carriers.
8. application of the polypeptide described in claim 1 or 2 in the medicine for preparing treatment prostatic disorders.
9. the application described in claim 8, wherein the prostatic disorders are related to secretion human 2 enzymes of kallikrein
Disease.
10. the application described in claim 8, wherein the prostatic disorders are prostate cancers.
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| CN201710044577.6A CN106831953A (en) | 2017-01-19 | 2017-01-19 | A kind of polypeptide and its application |
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| CN201710044577.6A CN106831953A (en) | 2017-01-19 | 2017-01-19 | A kind of polypeptide and its application |
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Family
ID=59120963
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108484734A (en) * | 2018-02-28 | 2018-09-04 | 北京化工大学 | A kind of polypeptide with anti-tumor activity and its application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085635A (en) * | 2015-09-07 | 2015-11-25 | 天津药物研究院有限公司 | Preparation method and application of breast cancer targeted peptide conjugated doxorubicin |
| CN106220735A (en) * | 2015-09-11 | 2016-12-14 | 中山大学 | A kind of preparation and application of cathepsin B activation type targeting anti-tumor polypeptide |
-
2017
- 2017-01-19 CN CN201710044577.6A patent/CN106831953A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085635A (en) * | 2015-09-07 | 2015-11-25 | 天津药物研究院有限公司 | Preparation method and application of breast cancer targeted peptide conjugated doxorubicin |
| CN106220735A (en) * | 2015-09-11 | 2016-12-14 | 中山大学 | A kind of preparation and application of cathepsin B activation type targeting anti-tumor polypeptide |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108484734A (en) * | 2018-02-28 | 2018-09-04 | 北京化工大学 | A kind of polypeptide with anti-tumor activity and its application |
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Application publication date: 20170613 |