CN105906691A - Eph kinase polypeptide inhibitor and application thereof - Google Patents
Eph kinase polypeptide inhibitor and application thereof Download PDFInfo
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- CN105906691A CN105906691A CN201610523440.4A CN201610523440A CN105906691A CN 105906691 A CN105906691 A CN 105906691A CN 201610523440 A CN201610523440 A CN 201610523440A CN 105906691 A CN105906691 A CN 105906691A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
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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
Abstract
The invention belongs to the technical field of biology and specifically discloses an Eph kinase polypeptide inhibitor and application thereof. The polypeptide disclosed by the invention is as shown in formula I. The polypeptide disclosed by the invention has a highly efficient inhibiting effect on Eph kinase, has higher stability and can be used for preparing medicine for treating or preventing Eph kinase abnormal expression related diseases. The polypeptide disclosed by the invention inhibits formation of tumor vasculature through inhibiting abnormal expression of Eph kinase, thereby playing the effect of anti-tumor growth, and the polypeptide can be used for preparing medicine for treating cancer.
Description
Technical field
The invention belongs to biological technical field, specifically Eph kinase polypeptide inhibitor and application thereof.
Background technology
Angiogenesis is one and relates to newly-generated angioblast and be gathered into primitive vessel clump, then experiences increasing
Grow, migrate, germination etc. is rebuild and is formed new blood vessel and the process of vasoganglion.Under physiology or pathological conditions,
Angiogenesis and fetal development growth, inflammatory reaction, wound healing, tumor diabetic retinal
The processes such as life are relevant.Angiogenesis is one of essential condition of growth and metastasis of tumours, and the most only tumor is raw
The long oxygen that necessity is provided and nutrient substance, and can be with excretion metabolism product.Therefore antineoplastic vascular is raw
Become the New Policy of oncotherapy.
Receptor tyrosine kinase Eph and part ephrins thereof occurs and neurodevelopment in tissue repair, tumor
Etc. the regulation and control of multiple biological processes are played the part of important role.14 kinds are had at present in mammalian body
The Eph receptor known and 8 kinds of ephrin parts.Similar to other most of tyrosine kinase receptors, Eph
Receptor family belongs to membrane cell surface molecule, is made up of three parts, i.e. born of the same parents' outer ligand binding domain, cross-film
District and intracellular region.Eph receptor family is divided into according to the difference of its part, the similarity film combination of structure
Two big classes: 9 kinds of EphA receptors can be combined with 5 kinds of A class ephrin parts, 5 kinds of EphB receptors can be with
3 kinds of B classes ephrin combine.The ephrin land of Eph receptor, at the aminoterminal of albumen, is connected to half Guang
Propylhomoserin enrichment region and two fibronectin Group III districts, then contain the tyrosine kinase of receptor in cell membrane
District.EphA receptoroid can be connected on cell membrane by the way of glycosyl-phosphatidyl inositol (GPI) grappling,
EphB receptoroid is then fixed on cell membrane by wearing membrane structure.
Recent study finds the receptor tyrosine kinase Eph and part ephrins thereof the life primitive vessel clump
Become and the structure of Subsequent vessel net plays an important role.Research finds that Eph kinases is multiple swollen
Overexpression in oncocyte such as breast carcinoma, colon cancer, bladder cancer etc., the suppression kinase whose activity of Eph is permissible
Suppress tumor vascular generation, thus reach the effect of neoplasm growth.
There is no suppression ephrin part and the combination of Eph receptor at present and play suppression Eph kinase activity
Small molecule, anti-tumor drug.Therefore, exploitation high activity and the kinase whose polypeptide of suppression Eph of high stability
Quasi-molecule becomes the focus of anti-tumor angiogenesis drug exploitation.The invention provides a kind of activity good steady
Qualitative high Eph kinase polypeptide inhibitor, preferably to meet the market demand.
Summary of the invention
It is an object of the present invention to provide the stable kinase whose polypeptide of suppression Eph and application thereof.
For realizing the purpose of the present invention, the present invention adopts the following technical scheme that
Polypeptide shown in a kind of formula Ι,
Or its isomer, diastereomer, enantiomer, pharmaceutically acceptable salt and its prodrug, wherein:
A is independently selected from Tyr, Ser or Thr；
B is independently selected from Asn, Asp, Glu or Gln；
C is independently selected from Ile, Leu, Phe or Val；
D is independently selected from His or Arg；
E is independently selected from Ala, Ile, Leu, Phe or Val；
X1For F-G fragment or amido link, F and G is respectively and independently selected from as Ala, Ile, Leu, Phe
Or Val.
Polypeptide of the present invention refers to combine the compound formed, formula Ι by covalent peptide bonds between aminoacid
From left to right it is expressed as from polypeptide N end to C end.Described aminoacid includes natural and alpha-non-natural amino acid.
Natural amino acid refers to the common main aminoacid for forming peptide and protein.Non-natural amino
Acid refers to that other are obtained by route of synthesis or derive from the aminoacid of natural resources.
Present invention also offers and have by replacing in the aminoacid sequence of polypeptide of the present invention, lacking
Or add the aminoacid sequence obtained by one or more amino acid residue and there is Eph kinase inhibiting activity
Polypeptide.Described aminoacid replacement, lack or add, can enter in any site of aminoacid sequence
OK, as long as the polypeptide with improved aminoacid sequence has Eph kinase inhibiting activity.Similar
, the amino acid whose number being replaced, lack or adding also is arbitrary, as long as having improved ammonia
The polypeptide of base acid sequence has Eph kinase inhibiting activity.
Present invention also offers the end modified gained of aminoacid sequence having by polypeptide of the present invention
To aminoacid sequence and there is the polypeptide of Eph kinase inhibiting activity.Polypeptide does not connect chemical group,
Then N-terminal is amino group, and C-terminal is carboxyl or amide group.Described polypeptide can be at its N-terminal
Or C-terminal connects chemical group and carries out end modified.Described modification can be phosphorylation, amidatioon, carbonyl
Base, alkylation, esterification or ubiquitination etc..
In some embodiments, it is provided that have by the aminoacid sequence N at polypeptide of the present invention
End is phosphorylated, amidatioon, carbonylation, ubiquitination, aminoacid obtained by pegylation
Sequence and there is the polypeptide of Eph kinase inhibiting activity.The most different compounds and the aminoacid sequence of polypeptide
The amino of N-terminal forms covalently bound group, includes formylated, acetylation, benzene as being acylated group
Formylated, trifluoroacetyl amination and succinylation etc., as long as it is many to have improved aminoacid sequence
Peptide has Eph kinase inhibiting activity.
In some embodiments, it is provided that have by the aminoacid sequence C at polypeptide of the present invention
End is partially alkylated or alkylated, be esterified modification obtained by aminoacid sequence and to have Eph kinase inhibiting activity many
Peptide.The most different compounds forms covalently bound base with the carboxyl of the aminoacid sequence C-terminal of polypeptide
Group, the carboxyl of compound Yu C-terminal as having the group such as the tert-butyl group, phenyl forms covalently bound base
Group.
In being embodied as embodiment, the invention provides and have shown in one of SEQ ID NO:1～6
Aminoacid sequence.
Present invention also offers and have by aminoacid multiple in the aminoacid sequence of polypeptide of the present invention
Aminoacid sequence obtained by residue cyclisation and there is the polypeptide of Eph kinase inhibiting activity.Described amino
Acid cyclisation can be carried out in any site of aminoacid sequence, as long as having the aminoacid sequence after cyclisation
Polypeptide has Eph kinase inhibiting activity.
Wherein, it is preferred that described cyclisation is formed by disulfide bond or amido link.
Present invention also offers and have by connecting fluorescence mark in the aminoacid sequence of polypeptide of the present invention
Note or radioactive label and there is the polypeptide of Eph kinase inhibiting activity.
Described fluorescent labeling can be connected to the arbitrary free ammonia in the N-terminal of polypeptide or peptide side chain
On base, as shown in Formula II-V, as long as the polypeptide with the aminoacid sequence after fluorescent labeling has Eph and swashs
Enzyme inhibition activity.
Preferably, described fluorescent labeling is FITC or FAM.
Present invention provides the DNA molecular encoding polypeptide of the present invention.Degeneracy due to codon
Property, a variety of nucleotide sequence that can encode specific polypeptide of the present invention can be there is.For compiling
The DNA molecular of code polypeptide of the present invention, those skilled in the art can utilize existing known easily
Method manufacture synthesis.Such as, by selecting the aminoacid corresponding to the aminoacid sequence designed by composition
The codon of residue, can be readily determined and provide the DNA of the aminoacid sequence corresponding to polypeptide to divide
Son.
Present invention also offers a kind of pharmaceutical composition, containing polypeptide of the present invention.
In some embodiments, described pharmaceutical composition also includes pharmaceutically acceptable adjuvant.As collapsed
Solve agent, lubricant, emulsifying agent, binding agent etc..
Experiment shows, polypeptide of the present invention has efficient inhibitory action to Eph kinases, and stable
Property is higher, can be used for the treatment of Eph kinases unconventionality expression relevant disease.Therefore the invention provides described
Polypeptide application in preparation treatment or prevention Eph kinases unconventionality expression relevant disease medicine.
Wherein, described Eph kinases unconventionality expression relevant disease is the destruction of cell breeding disease, angiogenesis
Caused, associated or adjoint disease, nervous system disease.
Cell breeding disease of the present invention is specially cancer.
Cancer of the present invention can be outside human primary gastrointestinal cancers class such as gastric cancer, esophageal carcinoma, carcinoma of small intestine, hepatocarcinoma, liver
Cancer of biliary duct, gastrointestinal class cancerous tumour, carcinoma of gallbladder.Apparatus urogenitalis cancer class, such as carcinoma of testis, carcinoma of penis,
Prostatitis cancer class；Gynecological cancer class such as ovarian cancer, cervical cancer, cancer of vagina, pudendum cancer, uterus/endometrium
Cancer, gestational trophoblastic tumor, carcinoma of fallopian tube, sarcoma of uterus；Head and tumor colli class include oral cancer,
Lip cancer, glandula cancer, larynx cancer, hypopharyngeal cancer, positive pharyngeal cancer, rhinocarcinoma, nasal sinus cancer, nasopharyngeal carcinoma；Leukemia class
As leukemia of children, acute lymphatic leukemia, acute myelogenous leukemia, chronic lymphatic leukemia,
Chronic lymphocytic leukemia, hair-like cell leukemia, acute promyelocytic leukemia, plasma cell
Property leukemia, bone marrow cancer, the bad syndrome of blood disorder such as bone marrow differentiation, myeloproliferative disease,
Aplastic anemia, Fanconi anemia, idiopathic macroglobulinemia disease；Pulmonary carcinoma class such as minicell lung
Cancer, nonsmall-cell lung cancer；Lymphatic cancer class such as Hodgkin, non Hodgkin lymphom, skin-type T-
Cell lymphoma, Peripheral T-cell lymphoma, AIDS related lymphoma；Cancer eye class such as retina is female
Glucagonoma, uveal；Skin carcinoma class such as melanoma, nonmelanoma skin cancer, prunus mume (sieb.) sieb.et zucc.
Ke Er cell carcinoma, soft tissue sarcoma class include child's soft tissue sarcoma, adult soft tissue sarcoma, Ka Boxi
Sarcoma；Urinary system cancer such as renal carcinoma, wilms' tumor, bladder cancer, carcinoma of urethra and transitivity is thin
Born of the same parents' cancer, osteocarcinoma class such as Ewing sarcoma, osteosarcoma, chondrosarcoma and cohorts, brain and cns tumor
Such as acoustic neuroma, neuroblastoma, neuroglia tumor and other cerebral tumors, tumor of spinal cord, breast carcinoma,
Colorectal carcinoma, progressive stage colorectal adenocarcinoma；Endocrine cancer class such as adrenocortical carcinoma, pancreas cancer,
Hypophysis cerebri cancer, thyroid carcinoma, parathyroid gland cancer, thymic carcinoma, multiple endocrine neoplasm.
Preferably, described cancer be breast carcinoma, colon cancerous protuberance, bladder cancer, ovarian cancer, carcinoma of prostate,
Melanoma, mesothelioma etc..
Polypeptide of the present invention can be natural polypeptides, recombinant polypeptide or synthesis polypeptide.The system of described polypeptide
Preparation Method is the present invention do not limit, as long as polypeptide of the present invention can be obtained.Some embodiment party
In case, can be by being chemically synthesized, as by solid-phase synthesis, liquid phase synthesizing method or solid phase liquid phase
The method synthesis combined.
As shown from the above technical solution, the invention provides the suppression kinase whose polypeptide of Eph.Of the present invention
Polypeptide has efficient inhibitory action to Eph kinases, and stability is higher, can be used for preparation treatment or
Prevention Eph kinases unconventionality expression relevant disease medicine.Polypeptide of the present invention is kinase whose by suppression Eph
Unconventionality expression, suppresses tumor vascular generation, thus reaches the effect of neoplasm growth, can be used for preparing
The medicine for the treatment of cancer.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to reality
Execute the required accompanying drawing used in example or description of the prior art to be briefly described.
Fig. 1 the invention provides polypeptide compound and ephrin-B1AP be incorporated into the Fc section albumen of EphB4
Activity suppression curve and calculated IC50Value；Wherein figure a is compound N I-E-00009 pair
Ephrin-B1AP is incorporated into the activity suppression curve of the Fc section albumen of EphB4 and calculated IC50
Value (2.94nM), figure b is the Fc that compound N I-E-00005 is incorporated into EphB4 to ephrin-B1AP
The activity suppression curve of section albumen and calculated IC50Value (1.4nM), figure c is compound
NI-E-00011 is incorporated into activity suppression curve and the meter of the Fc section albumen of EphB4 to ephrin-B1AP
The IC obtained50Value (2.85nM)；
Fig. 2 the invention provides polypeptide compound and ephrin-B2AP be incorporated into the Fc section albumen of EphB4
Activity suppression curve and calculated IC50Value；Wherein figure a is compound N I-E-00009 pair
Ephrin-B2AP is incorporated into the activity suppression curve of the Fc section albumen of EphB4 and calculated IC50Value
(9.4nM), figure b is the Fc section egg that compound N I-E-00010 is incorporated into EphB4 to ephrin-B2AP
White activity suppression curve and calculated IC50Value (40 μMs), figure c is compound N I-E-00005
Ephrin-B2AP is incorporated into the activity suppression curve and calculated of the Fc section albumen of EphB4
IC50Value (30nM)；
Fig. 3 the invention provides polypeptide compound in human lung adenocarcinoma cell (LU1250) conditioned medium not
Content with time period undegradable polypeptide；Wherein figure a compound N I-E-00005 and compound
NI-E-00009 the containing of different time sections in 4 hours in human lung adenocarcinoma cell (LU1250) conditioned medium
Amount, b is thin at human lung adenocarcinoma with matched group (NI-E-00013+ aprotinin) for compound N I-E-00013 for figure
The content of different time sections in 6 hours in born of the same parents (LU1250) conditioned medium, figure c is compound
NI-E-00009 and matched group (NI-E-00009+ aprotinin) and compound N I-E-00012 and matched group
(NI-E-00012+ aprotinin) in human lung adenocarcinoma cell (LU1250) conditioned medium in 210min not
Content with the time period.
Detailed description of the invention
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Describe, it is clear that described embodiment is only a part of embodiment of the present invention rather than all wholely
Embodiment.Based on the embodiment in the present invention, those of ordinary skill in the art are not making creativeness
The every other embodiment obtained under work premise, broadly falls into the scope of protection of the invention.In order to enter one
Step understands the present invention, and below in conjunction with specific embodiment, the present invention will be described in detail.As without special theory
Bright, the experiment reagent involved by the embodiment of the present invention is commercially available prod.Wherein portion of reagent title and contracting
That writes is corresponding as follows: DMF:N, dinethylformamide；DCM: dichloromethane；DIC:N, N-
DIC；HOBt:1-hydroxy benzo triazole；PIP: piperidines；Boc: tertbutyloxycarbonyl；
Trt: trityl；TBu: the tert-butyl group.
Embodiment 1 polypeptide compound of the present invention and synthesis thereof
The aminoacid sequence of Eph kinase polypeptide inhibitor NI-E-00005 is:
Tyr-Asn-Tyr-Leu-Phe-Ser-Pro-Asn-Gly-Pro-Ile-Ala-His-Ala-Trp-NH2(SEQ ID
NO:5), synthesize with following route.
1, the synthesis of Fmoc solid-phase synthesis is used.
Weigh Rink Amide-MBHA resin that 714mg substitution degree is 0.28mmol/g in solid state reaction
In device, add the swelling 30min of DCM, be subsequently added 20%PIP/DMF solution deprotection 2 times, the
5min, for the second time a 20min.Wash repeatedly with DCM Yu DMF respectively, drain.Anti-to solid phase
Answer and device add Fmoc-Trp (Boc)-OH (426mg, 1mmol), HOBt (135mg, 1mmol),
DIC (126mg, 1mmol), DMF (6mL), room temperature reaction 2h.Resin washing is drained, obtains Fmoc-
Trp (Boc)-Rink Amide-MBHA resin.Add 20%PIP/DMF solution deprotection 2 times, the
5min, for the second time a 20min, remove Fmoc protection group.3 are washed respectively with DCM Yu DMF
Secondary, obtain NH2-Trp (Boc)-Rink Amide-MBHA resin.
Addition Fmoc-Ala-OH (311mg, 1mmol) in solid phase reactor, HOBt (135mg,
1mmol), DIC (126mg, 1mmol), DMF (6mL), room temperature reaction 2h.Coupling uses after completing
Kaiser tests detection, detect by after carry out next step reaction.Resin washing is drained, to obtain final product
Fmoc-Ala-Trp (Boc)-Rink Amide-MBHA resin.The remaining amino of coupling the most successively
Acid, obtains side chain full guard polypeptide resin.
Add in solid phase reactor freezing 8mL lysate (volume ratio is trifluoroacetic acid: diphenyl sulfide:
Water=95:2.5:2.5), room temperature reaction 2h.Cracking reaction terminates, filtration resin, dichloromethane washing resin,
Merging filtrate and washing liquid, concentrated by rotary evaporation.Add the freezing diethyl ether solution of 10mL, separate out white precipitate, from
Collect white solid after the heart, be vacuum dried to obtain thick peptide 95mg.Thick peptide uses preparation HPLC purification, will
The liquid freezing prepared is dried to obtain target compound MS (m/z): 1749.79 [M+H]+。
Embodiment 2 polypeptide compound of the present invention and synthesis
The aminoacid sequence of Eph kinase polypeptide inhibitor NI-E-00013
Tyr-Asn-Tyr-Leu-Phe-Ser-Pro-Asn-Gly-Pro-Ile-Ala-His-Ala-Trp-Ahx-Lys(Biotin)
-NH2(SEQ ID NO:6), synthesizes with following route.
1, the synthesis of Fmoc solid-phase synthesis is used.
Weigh Rink Amide-MBHA resin that 357mg substitution degree is 0.28mmol/g in solid state reaction
In device, add the swelling 30min of DCM, be subsequently added 20%PIP/DMF solution deprotection 2 times, point
Fan Ying 5min and 20min.Wash repeatedly with DCM Yu DMF respectively, drain.To solid phase reactor
Middle addition Fmoc-Lys (Biotin)-OH (297.4mg, 0.5mmol), HOBt (67.5mg, 0.5mmol),
DIC (63mg, 0.5mmol), DMF (4mL), room temperature lucifuge reaction 2h.Resin washing is drained, i.e.
Obtain Fmoc-Lys (Biotin)-Rink Amide-MBHA resin.It is subsequently added 20%PIP/DMF solution to take off
Protect 2 times, reaction 5min and 20min respectively.Wash 3 times with DCM Yu DMF respectively,
NH2-Lys (Biotin)-Rink Amide-MBHA resin.
Addition Fmoc-6-Ahx-OH (176.7mg, 0.5mmol) in solid phase reactor, HOBt (67.5mg,
0.5mmol), DIC (63mg, 0.5mmol), DMF (4mL), room temperature lucifuge reaction 2h.Coupling completes
Rear use Kaiser reagent detect, detect by after carry out next step reaction.Resin washing is drained, i.e.
Obtain Fmoc-Ahx-Lys (Biotin)-Rink Amide-MBHA resin.Coupling is remaining the most successively
Aminoacid, obtains side chain full guard polypeptide resin.
Add in solid phase reactor freezing 6mL lysate (volume ratio is trifluoroacetic acid: diphenyl sulfide:
Water=95:2.5:2.5), room temperature lucifuge reaction 2h.Cracking reaction terminates, and filters resin, and dichloromethane washs
Resin, merging filtrate and washing liquid, concentrated by rotary evaporation.Add the freezing diethyl ether solution of 10mL, separate out white heavy
Forming sediment, collected after centrifugation white solid, lyophilization obtains thick peptide 101mg.Thick peptide uses preparation HPLC
Purification, is dried to obtain target compound MS (m/z): 2217.41 [M+H] by the liquid freezing prepared+。
Embodiment 3 polypeptide compound of the present invention
The aminoacid sequence of Eph kinase polypeptide inhibitor is as shown in table 1, according to embodiment 1 or embodiment 2
Described method synthesizes.
Table 1Eph kinase polypeptide inhibitor
Embodiment 4 biologic activity
1, the mensuration to EphB4 enzymatic activity
The Fc section of the 1mg/mL EphB4 being diluted in TBST is added in ELISA 96 orifice plate reacting hole
Albumen, hatches 1 hour under the conditions of room temperature lucifuge.Coated with TBST wash buffer EphB4Fc
After 96 orifice plates, add the polypeptide of variable concentrations and different volumes in each reacting hole includes ephrin-B2
The cell culture medium of AP, making cumulative volume is 50uL, hatches 1 hour under the conditions of room temperature lucifuge.
With TBST wash buffer reacting hole, remove unconjugated polypeptide and ephrin, with 1mM
PNPP (4-nitrophenyl sodium phosphate salts) is substrate, and excitation wavelength is 485nm, a length of 520nm of transmitted wave
Detecting its fluorescent value in microplate reader, GraphPad Prism4 mapping software calculates each polypeptide pair
EphB4 enzymatic activity i.e. IC50。IC50It is defined as suppressing the chemical combination required for the 50% of Eph enzymatic activity
Substrate concentration.Result is shown in Fig. 1, Fig. 2 and table 2.
2, vitro stability test
Take human lung adenocarcinoma cell (LU1250) to use containing 10% hyclone, penicillin and streptomycin
RPMI 1640 culture medium carries out cellar culture.The test-compound that concentration is 25nM is joined people's lung
In the conditioned medium of adenocarcinoma cell, cultivate 1-4 days for 37 DEG C.Collect the cell culture medium of different time sections also
With the dilution proportion of 1:10 in ELISA hole.ELISA is used to measure undegradable many
The content of peptide.Result such as Fig. 3 and Biao 2.
Vitro enzyme activity IC of table 2 the compounds of this invention50And stability test result
Note: n/d is for being not detected by；N/a is inapplicable
Fig. 1 result display SEQ ID NO:1,3,5 couple ephrin-B1AP is incorporated into the Fc of EphB4
The activity suppression curve of section albumen and calculated IC50Value.Fig. 2 result display SEQ ID NO 1,
2, the 5 couples of ephrin-B2AP are incorporated into the activity suppression curve of Fc section albumen of EphB4 and calculate
The IC arrived50Value.Fig. 3 result display SEQ ID NO 1,5,6 is at human lung adenocarcinoma cell (LU1250)
In conditioned medium the content of the undegradable polypeptide of different time sections and add after aprotinin 1,5,6
Content (the sun of the undegradable polypeptide of different time sections in human lung adenocarcinoma cell (LU1250) conditioned medium
Property comparison).
The above results shows that the compounds of this invention has certain inhibitory action to Eph kinases, particularly
Polypeptide shown in SEQ ID NO 4,5,6 is possible not only at low concentration suppression Eph receptor and its part ephrins
Combination, and there is higher stability, thus the most preferably play biological effect.
Claims (11)
1. a polypeptide shown in formula Ι,
Or its isomer, diastereomer, enantiomer, pharmaceutically acceptable salt and its prodrug, wherein:
A is independently selected from Tyr, Ser or Thr；
B is independently selected from Asn, Asp, Glu or Gln；
C is independently selected from Ile, Leu, Phe or Val；
D is independently selected from His or Arg；
E is independently selected from Ala, Ile, Leu, Phe or Val；
X1For F-G fragment or amido link, F and G is respectively and independently selected from as Ala, Ile, Leu, Phe
Or Val.
2. have by replacing in the aminoacid sequence of polypeptide described in claim 1, lack or adding one
Aminoacid sequence obtained by individual or multiple amino acid residue and there is the polypeptide of Eph kinase inhibiting activity.
3. have be phosphorylated by the aminoacid sequence N-terminal at polypeptide described in claim 1, amide
Aminoacid sequence obtained by change, carbonylation, ubiquitination, pegylation, or C-terminal is by alkane
Aminoacid sequence obtained by base, esterification, amidatioon modification, or it is former to connect N in amino acid whose peptide bond
Aminoacid sequence obtained by the methylated modification of son, and there is the polypeptide of Eph kinase inhibiting activity.
4. according to polypeptide described in claim 1-4, it is characterised in that have one of SEQ ID NO:1～6
Shown aminoacid sequence.
5. have and be cyclized by multiple amino acid residues in the aminoacid sequence of polypeptide described in claim 1
Obtained by aminoacid sequence and there is the polypeptide of Eph kinase inhibiting activity, the most described cyclisation is by two
Sulfide linkage or amido link are formed.
6. have by connecting fluorescent labeling or radiation in the aminoacid sequence of polypeptide described in claim 1
Property labelling and have the polypeptide of Eph kinase inhibiting activity, the most described fluorescent labeling is FITC or FAM.
7. a pharmaceutical composition, containing the polypeptide described in claim 1-6 any one.
Pharmaceutical composition the most according to claim 7, it is characterised in that also include pharmaceutically acceptable
Adjuvant.
9. polypeptide described in claim 1-6 any one or the medicine described in claim 7-8 any one
Compositions application in preparation treatment or prevention Eph kinases unconventionality expression relevant disease medicine.
Purposes the most according to claim 9, it is characterised in that described Eph kinases unconventionality expression
Relevant disease is cell breeding disease, angiogenesis is caused, associate or adjoint disease, nervous system
Disease.
11. purposes according to claim 10, it is characterised in that described cell breeding disease is cancer
Disease, the most described cancer be human primary gastrointestinal cancers, apparatus urogenitalis cancer, gynecological cancer, head and tumor colli, leukemia,
Blood disorder, pulmonary carcinoma, lymphatic cancer, cancer eye, skin carcinoma, urinary system cancer, osteocarcinoma, brain and CNS are swollen
Tumor, tumor of spinal cord, breast carcinoma, colorectal carcinoma, progressive stage colorectal adenocarcinoma, endocrine cancer class.
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| CN113945670A (en) * | 2021-09-30 | 2022-01-18 | 上海中科新生命生物科技有限公司 | Kinase enrichment detection method based on chemical proteomics |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113945670A (en) * | 2021-09-30 | 2022-01-18 | 上海中科新生命生物科技有限公司 | Kinase enrichment detection method based on chemical proteomics |
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