CN108640973A - The polypeptide and its dimer of a kind of targeting Syntenin albumen and application - Google Patents
The polypeptide and its dimer of a kind of targeting Syntenin albumen and application Download PDFInfo
- Publication number
- CN108640973A CN108640973A CN201810485601.4A CN201810485601A CN108640973A CN 108640973 A CN108640973 A CN 108640973A CN 201810485601 A CN201810485601 A CN 201810485601A CN 108640973 A CN108640973 A CN 108640973A
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- China
- Prior art keywords
- polypeptide
- syntenin
- dimer
- albumen
- affinity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
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- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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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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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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Abstract
The invention discloses a kind of polypeptide of targeting Syntenin albumen and its dimer and applications, belong to biomedicine technical field.The amino acid sequence of the polypeptide is Cys Asp Lys Φ Φ Val, and wherein Φ is 3 (2 naphthalene) L alanine.Polypeptide provided by the invention is to carry out amino acid to the natural polypeptides ligand screened to optimize to obtain, and significantly improves the affinity of its PDZ structural domain of connecting with Syntenin albumen.The dimer polypeptide that aforementioned polypeptides progress dimerization is obtained, it is up to 210nM with the affinity for PDZ structural domains of connecting, migration, proliferation, the invasion of a variety of cancer cells of Syntenin high expression can effectively be inhibited under 10uM concentration, other toxic side effects are not generated simultaneously, and the drug development for the Malignant tumor of bonal metastasis invasion for the treatment of Syntenin high expression provides mentality of designing.
Description
Technical field
The present invention relates to biomedicine technical fields, and in particular to a kind of polypeptide of targeting Syntenin albumen and its
Dimer and application.
Background technology
Syndecan binding protein Syntenin is the one kind being widely present in prokaryotes and eucaryote
Intracellular adaptin (adaptor proteins).Research finds that Syntenin albumen is high in a variety of high aggressive malignant tumours
Degree expression, and its expression is raised with the progress of metastases.
Syntenin is by N-terminal structural domain (N-terminal domain, NTD), two concatenated PDZ structural domains
(postsynaptic density protein, disc large and zonula occludens, PDZ) and C-terminal structural domain
(C-terminal domain, CTD) is formed.The PDZ structural domains of Syntenin albumen can be combined from the PDZ of different membrane receptor C-terminals
Motif (PDZ binding motif, PBM) is specifically bound, and the diversity of PDZ structural domain bind receptors results in syntenin
The diversity of function.
During tumour cell occurrence and development, PDZ structural domains and a series of ligand bindings simultaneously interact, and activate downstream
The phosphorylation of ERK1/2MAPK signal paths, and then a series of signal conduction, final upregulating matrix metalloproteinases MMP-2 occurs
Activity, degradation of cell epimatrix, promote tumor cell migration enter in the interstitial of surrounding.
Application No. is 200580028969.9 patent document disclose one kind can effectively inhibit PDZ structural domains interact
Small molecule, the compound overexpression DvI cancer cell in generate programmed cell death.
It is a potential drug target for inhibiting tumor invasion based on Syntenin albumen, Kegelman etc. is developed
A kind of micromolecular inhibitor class compound with syntenin Non-covalent bindings plays on inhibiting glioblastoma invasion
Good effect (Kegelman, T.P.eta1.Inhibition of radiation-induced glioblastoma
invasion by genetic and pharmacological targeting of MDA-9/
Syntenin.Proceedings of the National Academy of Sciences of the United States
Of America 2017,114, (2), 370-375.).But due to Syntenin albumen series connection PDZ structural domains contain there are one compared with
Big hydrophobic pocket structure, micromolecular compound due to molecular size limitation, it is relatively low with the affinity of albumen, only
20uM, therefore molecular concentration is up to 50uM in experiment, is otherwise just difficult to play inhibition, while small-molecule drug itself exists
The unfavorable factors such as metabolism is fast, toxicity is big.
In its natural state, the hydrophobic pocket of PDZ structural domains can interact therewith the C-terminal polypeptide combination of albumen, in turn
It forms complicated albumen composition and functions.With PDZ structural domains deeply grinding in multiple protein-protein-interacting
Study carefully, the inhibitor that a kind of and PDZ structural domains have high-affinity is provided, and then reaches and inhibit the pernicious swollen of Syntenin high expression
The purpose of tumor metastasis invasion, is those skilled in the art's problem to be solved.
Invention content
The purpose of the present invention is to provide a kind of inhibitor with high-affinity of targeting Syntenin albumen, lead to
It crosses and achievees the purpose that inhibit cancer cell migration, proliferation, invasion in conjunction with PDZ structural domains are connected in Syntenin albumen, to solve
Micromolecular inhibitor affinity is not high, the highly concentrated disadvantage of useful effect.
To achieve the above object, the present invention adopts the following technical scheme that：
Crystal structure (PDB ID of the present invention from Syntenin albumen PDZ structural domains and natural polypeptides ligand binding：
It 1w9o) sets out, filters out the natural polypeptides monomer of serial high-affinity.Research finds the second from the bottom of natural polypeptides ligand, three
A amino acid and the histidine 208 in PDZ hydrophobic pockets, valine Val 222 and phenylalanine Phe 213 have stronger
Interaction, therefore be the non-natural amino with strong hydrophobic residue by the second from the bottom of polypeptide ligand, three amino acid substitutions
Sour Φ：3- (2- naphthalenes)-l-Alanine (English is 3- (2-Naphthyl)-L-alanine), the polypeptide monomer after modification and string
The affinity of connection PDZ structural domains further enhances.
Therefore, the present invention provides a kind of polypeptide of targeting Syntenin albumen, the amino acid sequences of the polypeptide
It is 3- (2- naphthalenes)-l-Alanine for Cys-Asp-Lys- Φ-Φ-Val, wherein Φ.
The present invention also provides the polypeptides of the targeting Syntenin albumen to prepare Syntenin protein actives
Application in inhibitor.
To improve ability to function of the polypeptide as medicine, permeability enhanced portion is modified on the polypeptide
Point, such as cell-penetrating peptide R9, TAT, MAP, MPG, so that it is had transmembranal penetration, preferably, the polypeptide N-terminal is modified with rich essence
Propylhomoserin polypeptide R9.
The present invention using crosslinking agent by aforementioned polypeptides dimerization, the study found that compared to polypeptide monomer, dimer polypeptide with
The affinity of series connection PDZ structural domains greatly enhances.
Therefore, the present invention provides a kind of polypeptides of targeting Syntenin albumen to carry out dimerization formation
Dimer polypeptide.
The crosslinking agent is 1,11- dimaleoyl iminos -3,6,9- trioxy-s hendecane (1,11-bis
(maleimido) triethylene glycol) or 1,8- bis- (dimaleoyl imino) -3,6- dioxaoctanes (1,8-bis
(maleimido) diethylene glycol), difference lies in length differences for the two.
Preferably, according to Syntenin albumin crystal space structures, 1,11- dimaleoyl iminos -3,6,9- are selected
Trioxy- hendecane is as crosslinking agent.
Shown in the structural formula such as formula (I) of dimer polypeptide obtained,
Wherein Φ is 3- (2- naphthalenes)-l-Alanine.
Preferably, being modified with cell-penetrating peptide on the peptide chain of the dimer polypeptide.By being modified on dimer polypeptide
Permeability strengthening part, makes it have transmembranal penetration, improves its ability to function as medicine.Preferably, described
It is modified with rich arginine polypeptide on one of dimer polypeptide cysteine residues.The richness arginine polypeptide and described two
The amino terminal of homodimeric polypeptide cysteine residue is attached directly or by spacer.The rich arginine polypeptide by
9 L-type arginine residues compositions.
Shown in the structural formula such as formula (II) of the dimer polypeptide,
Wherein Φ is 3- (2- naphthalenes)-l-Alanine.
It is a further object to provide the applications of the dimer polypeptide.Dimer polypeptide after above-mentioned modification
The affinity of PDZ structural domains of connecting with Syntenin albumen is up to 210nM, and Syntenin albumen can be used as to connect PDZ structures
The inhibitor in domain and other protein-interactings, therefore, the present invention provides the dimer polypeptides to prepare Syntenin
Application in protein active inhibitor.
The present invention after PDZ structural domains, effectively presses down studies have shown that the dimer polypeptide combination Syntenin albumen is connected
The phosphorylation of the downstreams syntenin ERK1/2MAPK signal paths processed, thereby reduces the activity of matrix metalloproteinase MMP-2,
Therefore, the present invention provides the dimer polypeptides to prepare inhibition tumour cell matrix metalloproteinase MMP-2 activity
Application in the drug of up-regulation.
The present invention is studies have shown that the dimer polypeptide has high compatibility with Syntenin, under 10uM concentration
Migration, proliferation, the invasion of a variety of cancer cells of Syntenin high expression can effectively be inhibited, while not generating the secondary work of other poison
With.Therefore, the present invention provides the dimer polypeptides moves in the tumour cell for preparing inhibition Syntenin albumen height expression
It moves, be proliferated, the application in the drug of invasion.
Preferably, the tumour cell is Human cervical cancer cell lines, breast duct cancerous cell line.
The present invention also provides a kind of pharmaceutical compositions of Malignant tumor of bonal metastasis invasion that treating the expression of Syntenin albumen height
Object includes the dimer polypeptide (structural formula is as shown in Formula II) and pharmaceutically acceptable carrier of effective dose.
The effective quantity is the full dose for instigating treated object symptom to improve, and the improvement refers to dropping in the treatment
Negative effect caused by low or mitigation disease state.Active therapeutic ingredient is usually used in conjunction with pharmaceutical carrier, pharmaceutical carrier
It can be reasonably selected according to the common dosage form of existing polypeptide drug.
The advantageous effect that the present invention has：
The polypeptide of targeting Syntenin albumen provided by the invention is the natural polypeptides ligand by being obtained to screening
It carries out amino acid to optimize to obtain, significantly improves the affinity of its PDZ structural domain of connecting with Syntenin albumen.By aforementioned polypeptides
The dimer polypeptide that dimerization obtains is carried out, 210nM is up to the affinity for PDZ structural domains of connecting, is the first of hitherto reported
For the peptide inhibitor of Syntenin albumen.
Dimer polypeptide provided by the invention has high compatibility with Syntenin, can effectively press down under 10uM concentration
Migration, proliferation, the invasion of a variety of cancer cells of Syntenin high expression processed, while other toxic side effects are not generated, for treatment
The drug development of the Malignant tumor of bonal metastasis invasion of Syntenin high expression provides mentality of designing.
Description of the drawings
Fig. 1 is that the binding force of monomer polypeptide and syntenin PDZ1 characterizes, wherein (A) is p1, (B) is p2, and (C) is p3,
(D) it is p4.
Fig. 2 is that the binding force of monomer polypeptide and syntenin PDZ2 characterizes, wherein (A) is p1, (B) is p2, and (C) is p3,
(D) it is p4.
Fig. 3 is dimer polypeptide structure formula schematic diagram.
Fig. 4 be dimer polypeptide connect with syntenin PDZ structural domains affinity characterize, wherein (A) be p2-2, (B)
It is p2-4 for p2-3, (C), (D) is p3-3, and (E) is p3-4, and (F) is p4-4.
Fig. 5 be monomer polypeptide connect with syntenin PDZ structural domains affinity characterization, wherein (A) be p2, (B) be p3,
(C) it is p4.
Fig. 6 is that the monomer polypeptide of optimization is characterized with the affinity for PDZ structural domains of connecting, wherein (A) is p5, (B) is p6,
(C) it is p7, (D) is p8, and (E) is p9, and (F) is p10, and (G) is p11, and (H) is p12, and (I) is p13.
Fig. 7 be dimer polypeptide structure (A) after optimization and its with the affinity of PDZ structural domains of connect characterization (B)-(D)
(E)-(F) is characterized with the affinity of p13-13 and monomer PDZ1, PDZ2.
Fig. 8 is the syntenin expressions of different cancer cells, wherein (A) is mRNA level in-site testing result, (B) is
Western blot testing results.
Fig. 9 is the scar Healing Experiments of Hela cell lines, wherein (A) is pictorial diagram, (B) is data analysis figure.
Figure 10 is the phosphorylation testing result that dimer polypeptide inhibits ERK1/2MAPK signal paths, wherein (A) is
Weatem blot detect ERK expressions, and (B) is data analysis figure.
Figure 11, which is dimer polypeptide, reduces the Activity determination of matrix metalloproteinase MMP-2 as a result, wherein (A) is people's uterine neck
Cancerous cell line Hela cell lines, (B) are breast duct cancerous cell line MDA-MB-435, and (C) is breast adenocarcinoma cell system MDA-MB-
231。
Figure 12 is the proliferation testing result that dimer polypeptide inhibits syntenin high expression cancer cells, wherein (A) is people palace
Neck cancer Cell line Hela cell line, (B) are breast duct cancerous cell line MDA-MB-435, and (C) is breast adenocarcinoma cell system MDA-
MB-231。
Specific implementation mode
The present invention is further explained in the light of specific embodiments.
One, the synthesis of polypeptide and affinity characterize
Following four polypeptide sequences are chosen from document：RVAFFEEL (polypeptide epitope of Merlin albumen), TNEFYA
(polypeptide epitope of syndecan albumen), DKEYYV (polypeptide epitope of neurexin albumen), LEDSVF (interleukin-5
The polypeptide epitope of receptor α albumen).
For the ease of subsequent characterizations, FAM fluorophors are added in N-terminal, and using GS amino acid as fluorophor and function
The interval of amino acid region.
1, the synthesis of monomer polypeptide
1) Fmoc chemical solid phase Peptide systhesis is used, using Fmoc-wang resins, with 5 times of excessive fmoc-protected ammonia
Base acid and condensation reagent (HBTU/HOBt) synthesis polypeptide.
2) in the place addition cysteine C close to polypeptide N-terminal, sulfydryl (HS-) reactive group is provided.
3) after polypeptide N-terminal Fmoc deprotections, FAM fluorophors are connected with EDC/HOBt condensing agents.
4) polypeptide is cut off with 95%TFA from resin, is precipitated with ice ether, obtained preliminary polypeptide HPLC separation is pure
Change.
2, the synthesis of dimer polypeptide
The characteristics of being reacted with maleimide high specific using sulfydryl (HS-), by two polypeptide monomers respectively with horse
Carry out acid imide-PEG3 link object reactions, obtains polypeptide dimer.
3, polypeptide characterizes
The polypeptide isolated and purified through HPLC detects confirmation molecular weight with MALDI-TOF MS, and purity is verified with HPLC.
4, the external affinity of polypeptide and syntenin albumen PDZ tandem domains characterizes
(1) structure pGEX-Syntenin tandem PDZ, pGEX-Syntenin PDZ1, pGEX-Syntenin PDZ2
Prokaryotic expression plasmid
Syntenin tandem PDZ prokaryotic expression gene sequences are designed according to Syntenin tandem PDZ cDNA sequences
Row, and synthesized by related gene Synesis Company.
Obtain Syntenin tandem PDZ segments with digestion method, with PCR method obtain Syntenin PDZ1 and
Syntenin PDZ2 segments are used in combination in T4 ligases (Takara) connection prokaryotic expression plasmid pGEX-6P-1, and sequence verification.
(2) E.Coli prokaryotic expressions and purifying protein Syntenin tandem PDZ, Syntenin PDZ1, pGEX
PDZ2
Verified correct plasmid conversion after choosing clone's verification correctly, is added in sterilizing in BL-21 competent cells
It is shaken overnight for 37 DEG C in 5mL LB culture mediums.It takes 4mL to be added in 400mL LB later and shakes OD for 37 DEG C600About 04-0.6 adds IPTG to lure
It leads, then shakes 18 hours for 16 DEG C.Centrifuge removes supernatant of bacteria solution, thalline ultrasonication later.
Using GST label purifying proteins, Protein S yntenin tandem PDZ, Syntenin PDZ1, pGEX are obtained
PDZ2 is simultaneously verified with SDS-PAGE.
(3) the external affinity of polypeptide and syntenin albumen PDZ structural domains characterizes
Freeze-drying polypeptide is dissolved in protein storage buffer solution, then by polypeptide, protein solution 0.1um PVDM membrane filtrations,
Remove molecule that may be present.Then the fluorescence polarization value of polypeptide solution is detected with sepectrophotofluorometer, later every time
A certain amount of albumen is added, is incubated after a certain period of time, detects the fluorescence polarization value of polypeptide-mixed liquid of protein, a series of differences are dense
The corresponding fluorescence polarization value of degree albumen does figure, can fit the affinity of polypeptide and albumen.
A. the external affinity characterization result institute as shown in Figure 1, Figure 2 of monomer polypeptide (table 1) and syntenin albumen PDZ structural domains
Show, and is summarised in table 2.
The sequence of 1 monomer polypeptide of table
| Monomer polypeptide | Sequence |
| p1 | FAMCGSYGSRVAFFEEL |
| p2 | FAMGCGSTNEFYA |
| p3 | FAMGCGSDKEYYV |
| p4 | FAMGCYGSLEDSVF |
The binding force of 2 monomer polypeptide of table and syntenin PDZ1 and PDZ2 structural domains
| Affinity Kd(μM) | p1 | p2 | p3 | p4 |
| Syntenin PDZ1 | 282±29 | 94±6 | 81±7 | 58±4 |
| Syntenin PDZ2 | 531±114 | 68±4 | 33±2 | 60±5 |
Experiment finds that natural monomers polypeptide ligand and the affinity of syntenin PDZ structural domains are medium on the weak side, with document report
Road is consistent.Due to weak an order of magnitude of binding force ratio p2, p3, p4 of p1 and syntenin PDZ, subsequent design synthesis with
With carrying out based on the functional sequence of p2, p3, p4.
B. maleimide-PEG3 is utilized to link object by monomer polypeptide p2, p3, p4 dimerization, dimer polypeptide structure is shown in
Fig. 3.Fluorescence polarization Experimental Characterization dimer polypeptide is connected the affinity of PDZ structural domains with syntenin, sees Fig. 4, and summarize
In table 3.
3 dimer polypeptide of table is connected the binding forces of PDZ structural domains with syntenin
With single phase ratio (Fig. 5), connect with the syntenin affinity of PDZ structural domains of dimer polypeptide substantially increases.
C. the optimization of polypeptide monomer sequence
By crystal structure analysis, monomer polypeptide ligand C-terminal is second from the bottom, three amino acid and the group in PDZ hydrophobic pockets
Propylhomoserin His 208, valine Val 222 and phenylalanine Phe 213 have stronger interaction.In preceding step experiment, p3 is more
Peptide monomer and the affinity for PDZ structural domains of connecting are most strong.Therefore, we based on p3 polypeptide sequences, C-terminal is second from the bottom,
Three amino acid (tyrosine Y) replace with the amino acid with different hydrophobic residues：Tryptophan W, phenylalanine F and non-day
Right amino acid Φ (3- (2- naphthalenes)-l-Alanine (English is 3- (2-Naphthyl)-L-alanine) naphthyl-
Alanine), synthesis obtains p5-p13, and sequence is shown in Table 4.Affinity characterizes as shown in fig. 6, and summarizing in table 4.
The monomer polypeptide sequence and its summarized with the affinity for PDZ structural domains of connecting that table 4 optimizes
After replacing with phenylalanine F, monomer polypeptide (p8-p10) affinity is not much different with original polypeptide p3, thus it is speculated that be due to
F hydrophobic residues phenyl is similar to the hydroxyphenyl size of tyrosine Y；After replacing with tryptophan W, monomer polypeptide (p5-p7) affinity
It is more slightly higher than former polypeptide p3, thus it is speculated that be since the hydroxyphenyl of W hydrophobic residues indyl ratio Y is big；After replacing with non-natural amino acid Φ,
Monomer polypeptide (p11-p13) affinity is significantly improved than former polypeptide p3, thus it is speculated that is since Φ hydrophobic residue naphthalenes are with strongest
Hydrophobicity, therefore have very strong interaction with the hydrophobic pocket of PDZ structural domains.
D. the polypeptide dimerization body after optimizing connects PDZ structural domains with syntenin with high affinity
Monomer polypeptide dimerization can be greatly improved to the affinity of polypeptide and PDZ tandem domains, therefore I known to aforementioned
By monomer polypeptide p11, p12, p13 dimerization with higher affinity after optimization, sequential structure is shown in Fig. 7 (A).By Fig. 7
(B) dimerization known to-(F) significantly improves the affinity between polypeptide and PDZ structural domains of connecting, and is summarised in table 5, wherein
P13-13 affinity is up to 210nM, and 160 times are improved than former polypeptide p3 affinity.
Table 5p13-13 affinity characterizes
As a result, by a series of designs and synthesis, we have successfully obtained the dimer polypeptide p13-13 of high-affinity.
Two, polypeptide antitumous effect is detected in the different cancerous cell lines of syntenin high expression
1, the case where different cell line expression syntenin
TRIzol extracts total serum IgE in cell, is used in combination reverse transcription method to carry out specific amplification syntenin albumen, qPCR is used in combination
Quantification of mrna expression quantity.
By Human cervical cancer cell lines HeLa, breast duct cancerous cell line MDA-MB-435, breast adenocarcinoma cell system MDA-MB-
231 are laid on 10mm culture dishes.It covers with rear PBS to wash and cracked on ice with lysate, takes cracking supernatant, use Western
Blot experiment detection syntenin protein expression situations.If expression, albumen can be identified by specific syntenin antibody, and most
Band is showed eventually.
By mRNA expression and protein expression (see Fig. 8) it is found that HeLa and MDA-MB-435 high express syntenin, and mammary gland
Gland cell system MDA-MB-231 hardly expresses syntenin.
2, influence detection of the dimer polypeptide to cancer cell migration
Scar Healing Experiments Scratch wound migration assays：Hela cells are laid on 24 orifice plates, per hole 1
×105Cell is more after adding dimer polypeptide p13-13 (R9-dimer), cell-penetrating peptide R9 after cell-penetrating peptide R9 modifications to modify respectively
Peptide p13 (R9-mono) and control monomer polypeptide R9, acts on 20min by a concentration of 10 μM.With 10ul pipette tips in hole middle position
Standardized scar calculates cell migration length, the results are shown in Figure 9 respectively in 0h and for 24 hours in micro- lower observation scar width.
The above results show that the 13-13 dimers polypeptide of 10uM can effectively inhibit syntenin high to express cancer cell
Migration.
3, influence of the dimer polypeptide for syntenin downstream signaling pathways.
The phosphorylation of ERK1/2MAPK signal paths：HeLa is laid on 10mm culture dishes, respectively plus after cell-penetrating peptide R9 modifications
Dimer polypeptide p13-13 (R9-dimer), cell-penetrating peptide R9 modification after polypeptide p13 (R9-mono) and compare monomer polypeptide
R9, acts on 20min by a concentration of 10 μM.It is cracked after cell pellet overnight culture, with the ERK of Western blot experimental verification phosphorylations
Expression, the results are shown in Figure 10.
The activity of matrix metalloproteinase MMP-2：HeLa is laid on 12 orifice plates, adds the dimerization after cell-penetrating peptide R9 modifications respectively
Polypeptide p13 (R9-mono) after body polypeptide p13-13 (R9-dimer), cell-penetrating peptide R9 modifications and control monomer polypeptide R9, concentration
It is 10 μM, acts on 20min.Supernatant is taken after cell incubation 48h, with matrix metalloproteinase MMP-2 enzyme linked immunosorbent assay reagents
Box detects the activity of MMP-2, as a result as shown in figure 11.
The above results show that dimer polypeptide effectively inhibits the phosphoric acid of the downstreams syntenin ERK1/2MAPK signal paths
Change, thereby reduces the activity of matrix metalloproteinase MMP-2.
4, influence of the dimer polypeptide for cancer cell multiplication
As shown in figure 12, dimer polypeptide effectively inhibits the proliferation of syntenin high expression cancer cells.
Sequence table
<110>The first maternity and infant health institute, Shanghai City
<120>The polypeptide and its dimer of a kind of targeting Syntenin albumen and application
<160> 13
<170> SIPOSequenceListing 1.0
<210> 1
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> UNSURE
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for 3-(2-Naphthyl)-L-alanine
<220>
<221> UNSURE
<222> (6)..(6)
<223> The 'Xaa' at location 5 stands for 3-(2-Naphthyl)-L-alanine
<220>
<221> UNSURE
<222> (5)..(5)
<223> The 'Xaa' at location 5 stands for Gln, Arg, Pro, or Leu.
<220>
<221> UNSURE
<222> (6)..(6)
<223> The 'Xaa' at location 6 stands for Gln, Arg, Pro, or Leu.
<400> 1
Cys Asp Lys Glu Xaa Xaa Val
1 5
<210> 2
<211> 14
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 2
Cys Gly Ser Tyr Gly Ser Arg Val Ala Phe Phe Glu Glu Leu
1 5 10
<210> 3
<211> 10
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 3
Gly Cys Gly Ser Thr Asn Glu Phe Tyr Ala
1 5 10
<210> 4
<211> 10
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 4
Gly Cys Gly Ser Asp Lys Glu Tyr Tyr Val
1 5 10
<210> 5
<211> 11
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 5
Gly Cys Tyr Gly Ser Leu Glu Asp Ser Val Phe
1 5 10
<210> 6
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 6
Cys Asp Lys Glu Tyr Trp Val
1 5
<210> 7
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 7
Cys Asp Lys Glu Trp Trp Val
1 5
<210> 8
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 8
Cys Asp Lys Glu Trp Tyr Val
1 5
<210> 9
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 9
Cys Asp Lys Glu Phe Phe Val
1 5
<210> 10
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 10
Cys Asp Lys Glu Phe Trp Val
1 5
<210> 11
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<400> 11
Cys Asp Lys Glu Tyr Phe Val
1 5
<210> 12
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> UNSURE
<222> (6)..(6)
<223> The 'Xaa' at location 5 stands for 3-(2-Naphthyl)-L-alanine
<220>
<221> UNSURE
<222> (6)..(6)
<223> The 'Xaa' at location 6 stands for Gln, Arg, Pro, or Leu.
<400> 12
Cys Asp Lys Glu Tyr Xaa Val
1 5
<210> 13
<211> 7
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
Claims (10)
1. a kind of polypeptide of targeting Syntenin albumen, which is characterized in that the amino acid sequence of the polypeptide is Cys-
Asp-Lys- Φ-Φ-Val, wherein Φ are 3- (2- naphthalenes)-l-Alanine.
2. the polypeptide of targeting Syntenin albumen as described in claim 1 is preparing Syntenin protein active inhibitor
In application.
3. the dimer that a kind of polypeptide by targeting Syntenin albumen described in claim 1 carries out dimerization formation is more
Peptide.
4. dimer polypeptide as claimed in claim 3, which is characterized in that the structural formula such as formula (I) of the dimer polypeptide
It is shown,
Wherein Φ is 3- (2- naphthalenes)-l-Alanine.
5. dimer polypeptide as described in claim 3 or 4, which is characterized in that be modified on the peptide chain of the dimer polypeptide
Cell-penetrating peptide.
6. dimer polypeptide as claimed in claim 5, which is characterized in that structural formula such as formula (II) institute of the dimer polypeptide
Show,
Wherein Φ is 3- (2- naphthalenes)-l-Alanine.
7. such as claim 3-6 any one of them dimer polypeptides answering in preparing Syntenin protein active inhibitor
With.
8. as claim 3-6 any one of them dimer polypeptides are preparing inhibition tumour cell matrix metalloproteinase
Application in the drug of MMP-2 activity up-regulation.
9. as claim 3-6 any one of them dimer polypeptides are thin in the tumour for preparing inhibition Syntenin albumen height expression
Born of the same parents' migration is proliferated, the application in the drug of invasion.
10. application as claimed in claim 9, which is characterized in that the tumour cell is Human cervical cancer cell lines, mammary gland is led
Pipe cancerous cell line.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110330548A (en) * | 2019-06-28 | 2019-10-15 | 哈尔滨医科大学 | A kind of antineoplastic polypeptide and its application in preparation of anti-tumor drugs |
| WO2024000262A1 (en) * | 2022-06-29 | 2024-01-04 | 京东方科技集团股份有限公司 | Probe and kit for early diagnosis of diffuse large b-cell lymphoma |
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2018
- 2018-05-19 CN CN201810485601.4A patent/CN108640973B/en active Active
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| ANDERS BACH等: "A high-affinity, dimeric inhibitor of PSD-95 bivalently interacts with PDZ1-2 and protects against ischemic brain damage", 《PNAS》 * |
| GREMBECKA，J.等: "The Binding of the PDZ Tandem of Syntenin to Target Proteins", 《BIOCHEMISTRY》 * |
| TIMOTHY P. KEGELMAN等: "Inhibition of radiation-induced glioblastoma invasion by genetic and pharmacological targeting of MDA-9/Syntenin", 《PNAS》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110330548A (en) * | 2019-06-28 | 2019-10-15 | 哈尔滨医科大学 | A kind of antineoplastic polypeptide and its application in preparation of anti-tumor drugs |
| CN110330548B (en) * | 2019-06-28 | 2021-03-09 | 哈尔滨医科大学 | Anti-tumor polypeptide and application thereof in preparation of anti-tumor medicine |
| WO2024000262A1 (en) * | 2022-06-29 | 2024-01-04 | 京东方科技集团股份有限公司 | Probe and kit for early diagnosis of diffuse large b-cell lymphoma |
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| Publication number | Publication date |
|---|---|
| CN108640973B (en) | 2021-06-11 |
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