CN107129519A - Polypeptide fluorescence cyclization method and cyclized polypeptide based on metal iridium complex - Google Patents
Polypeptide fluorescence cyclization method and cyclized polypeptide based on metal iridium complex Download PDFInfo
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- CN107129519A CN107129519A CN201710308188.XA CN201710308188A CN107129519A CN 107129519 A CN107129519 A CN 107129519A CN 201710308188 A CN201710308188 A CN 201710308188A CN 107129519 A CN107129519 A CN 107129519A
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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
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/55—Fusion polypeptide containing a fusion with a toxin, e.g. diphteria toxin
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/18—Metal complexes
- C09K2211/185—Metal complexes of the platinum group, i.e. Os, Ir, Pt, Ru, Rh or Pd
Abstract
The invention discloses a kind of polypeptide fluorescence cyclization method and cyclized polypeptide based on metal iridium complex.Technical scheme mainly includes：Cyclometalated iridium coordination compound is provided, and two histidine residues at the Cyclometalated iridium coordination compound and the diverse location that is distributed in polypeptide peptide chain is occurred complexation reaction, polypeptide cyclisation is realized, and produce can by ultraviolet excitation green fluorescence；Wherein the chemical molecular formula of Cyclometalated iridium coordination compound is (LC^N)2Ir(Lsolv)2M, M may be selected from OTf-、PF6 -Or BF4 -Deng.Using Cyclometalated iridium coordination compound with 2 histidine residues in polypeptide peptide chain complexation reaction occurs for the present invention, complete polypeptide cyclisation, the fluorophor that can be excited is formed simultaneously, peptide molecule is set to be labeled fluorescence, technique is simply efficient, and reaction condition is gentle, and product property is stable, without obvious bio-toxicity, with wide biochemistry and medical application prospect.
Description
The application is the number of patent application submitted on March 6th, 2014 for 201410079377.0, entitled " be based on metal
The divisional application of the application for a patent for invention of the polypeptide fluorescence cyclization method and cyclized polypeptide of complex of iridium ", and require patent application
Number be 201310077634.2, patent application day be on March 12nd, 2013, it is entitled that " one kind is matched somebody with somebody based on histidine and metal iridium
The domestic priority of the Chinese invention patent application of the polypeptide cyclisation method that compound is combined ".
Technical field
It is particularly a kind of to be based on a class and polypeptide the present invention relates to a kind of novel polypeptide fluorescence cyclization method and cyclized polypeptide
In may be selected site two His-Bind metal complex of iridium, complete two histidines between peptide sequence cyclisation, together
Shi Shengcheng fluorophors, and the polypeptide fluorescence cyclization method combined based on histidine with metal iridium complex realized.
Background technology
In recent years, polypeptide and various small molecular proteins are extensive due to superior biocompatibility and plasticity
Treatment applied to a variety of diseases.As the executor of intracellular most of signal path, these crucial peptide molecules can be with
By regulating and controlling the signal path related to disease, there is provided the action target spot of medicine.In addition, as bioactive molecule, it is many
Peptide wide material sources, it is easy to synthesize, and different polypeptides can provide and wear film, target, a variety of functions such as treatment, be increasingly becoming and work as
For one of medical science and the important tool in life science field.
Cyclized polypeptide is widely distributed in nature, plant, animal, low marine organisms, microorganism, bacterium and germ etc.
All contain micro cyclized polypeptide.The content of these cyclic peptide is although low, but wherein much has obvious physiologically active, also therefore
Paid attention to by domestic and international many chemists, biologist and pharmacy man.The cyclisation of peptide is therefore as a kind of important polypeptide point
Sub- design method, cyclisation can reduce the free degree of ring group point, and the specific secondary structure of stable polypeptide.Research shows, limits many
The conformation of peptide can increase the affinity and selectivity combined with acceptor, and the properties under its cyclisation state are significantly better than that it
Linear condition.For example, RGD peptide (tripeptide sequence containing arginine-glycine-aspartic acid) can be with a variety of integrins specificity
With reference to can be effectively facilitated adhesion of the cell to biomaterial, reach the purpose of the plain high expressing cell of targeted integration.Study table
Bright, after it is cyclized, in stability, targeting and affinity everyway are substantially better than linear RGD.
Traditional linear polypeptide cyclization method has 2 kinds：One class is the condensation by amino and carboxyl, forms peptide bond.It is this
Traditional head and the tail cyclisation method is related to all or part of protection and deprotection strategy and needs condensation reagent progress correlative link anti-
Should, process is complicated, and is difficult to the extension of doing structure after cyclisation, it is impossible to do more complicated design and functionalization.It is another kind of by drawing
Enter disulfide bond (s-s) to be cyclized biologically active peptide.Generally first synthesis carries sulfhydryl protected cysteine (Cys) residue
Line style peptide precursor, then sloughs Cys protection group, is oxidized to cyclization after disulfide bond, or be directly oxidized to the Cys with protection group
For s-s keys, but S -- S is unstable under the reducing environment of natural biosystem, greatly limit this kind of bioactivity ring
Change further applying for polypeptide.
The content of the invention
It is a primary object of the present invention to provide a kind of cyclized polypeptide and based on metal iridium complex polypeptide fluorescence cyclisation
Method, its technique is simple, quick, easy to implement, and obtained product is the features such as have hypotoxicity, high stability, so as to overcome
Deficiency of the prior art.
For achieving the above object, present invention employs following technical scheme：
A kind of cyclized polypeptide, comprising mainly by Cyclometalated iridium coordination compound and the diverse location being distributed in polypeptide peptide chain
Two histidine residues at place pass through cyclized structure formed by complexation reaction.
A kind of polypeptide cyclisation method combined based on histidine with metal iridium complex, including：
Cyclometalated iridium coordination compound is provided, and makes the Cyclometalated iridium coordination compound and is distributed in polypeptide peptide chain
Complexation reaction occurs for two histidine residues at diverse location, realizes polypeptide cyclisation, and generation can be by the green of ultraviolet excitation
Color fluorescence, wherein, the N-terminal of the polypeptide peptide chain is free of histidine residues or contains protected histidine residues.
Specifically, the chemical molecular formula of the Cyclometalated iridium coordination compound is (LC^N)2Ir(Lsolv)2M, its structural formula
It is as follows：
Wherein, LC^NFor C^N bidentate ligands, LsolvFor solvent molecule.
Further, the C^N bidentate ligands can select but be not limited to 2- phenylpyridines, 1- phenyl isoquinolin quinolines, 2- phenyl
Benzothiophene, benzoquinoline, 4- methylphenylpyridiniums, 4,6- difluorophenyl pyridinatos, 2- (benzothiophene) pyridine, 2- phenylchinolines
With 2,5- diphenyloxazoles.
Further, the solvent molecule may be selected from but be not limited to water, acetonitrile, DMF or DMSO etc..
Further, M-It may be selected from but be not limited to trifluoromethane sulfonic acid root (OTf-), hexafluoro-phosphate radical (PF6 -), tetrafluoro boron
Acid group (BF4 -)。
Further, the complexation reaction is carried out in liquid phase environment, and the liquid phase environment is main by buffer solution system
Composition, the buffer solution system can select but be not limited to phosphate buffer (PBS), Tris buffer solutions, Bis-Tris buffer solutions,
MES buffer solutions or HEPES buffer solution.
Further, in the polypeptide peptide chain, two of complexation reaction occur with the Cyclometalated iridium coordination compound
0-5 any amino acid in addition to histidine is spaced between histidine, particularly preferably 2-3, interval is in addition to histidine
Any amino acid.
Further, the polypeptide can select but be not limited to RGD peptide etc..
Compared with prior art, advantages of the present invention at least that：
(1) the invention provides a kind of novel, easy, stable polypeptide cyclisation mode, it is by by Cyclometalated iridium coordination
Compound is combined with 2 histidine residues (Histidine) in polypeptide peptide chain, is cyclized the peptide sequence between histidine,
Realize fluorescence labeling simultaneously, even if also, polypeptide sequence it is different, can be with complete as long as being containing 2 histidine residues in peptide chain
Into the cyclisation of peptide sequence therebetween, course of reaction is quick, easy, and reaction condition is gentle, and cyclisation product is stable, irreversible,
It is non-toxic in biosystem, it is easy to proceed modification and the assembling process of complexity, analysis detection, fluorescence can be widely used in
The technical fields such as imaging.
(2) cyclization of the invention can be carried out in cushioning liquid at normal temperatures, conventional, it is not necessary to extra crosslinking examination
Agent, reaction condition is gentle, and operating procedure is simple, and course of reaction is quick, and reaction efficiency is high, with low cost；
(3) Cyclometalated iridium coordination compound used in the present invention does not possess before the histidine residues with polypeptide are combined
The compound formed in fluorescence, complexation process just has obvious Fluorescence Increasing, thus a step complete structure optimization (cyclisation) with
And the mark of fluorescence, while the fluorescence combination chemical characterization means (mass spectrum, UPLC) produced can accurately prove polypeptide cyclisation
Success or not, also, the present invention Cyclometalated iridium coordination compound also have preferable photostability, longer fluorescence lifetime
Etc. photophysics characteristic and chemical stability, it can be preserved for a long time with solid or solution form under normal conditions.
(4) present invention is in actual application process, assign polypeptide (e.g., RGD) cyclisation structure (Ir-HRGDH, this
The Ir systems Cyclometalated iridium coordination compound at place is write a Chinese character in simplified form) after, also reflect and linear condition (RGDHH-Ir, Ir herein
Implication is ibid) more preferable biological selectivity is compared, show that the inventive method can reach the optimization that traditional cyclisation means are completed
The effect of polypeptide structure and function.
Brief description of the drawings
Fig. 1 is a kind of structure of the cyclized polypeptide based on metal iridium complex in the more preferred embodiment of the present invention one
Schematic diagram, wherein, H is histidine, and X is amino acid, and n is the integer more than or equal to zero；
Fig. 2A-Fig. 2 D respectively show Land use models polypeptide A c-HA in embodiment 1nH-NH2(it is abbreviated as " HAnH ") (n=
0,1,2,3,4,5) product structure, the reaction of complexation reaction occur with Cyclometalated iridium coordination compound (being abbreviated as " Ir ", similarly hereinafter)
The stability collection of illustrative plates of (GSH=5mM) under speed, the fluorescence intensity and reducing environment of generation；
Fig. 3 is polypeptide A c-HRGDH-NH in 2-3 of the embodiment of the present invention2(being abbreviated as " HRGDH ") or Ac-HRGDH-
(KLAKLAK)2(it is abbreviated as " HRGDH-KLA " or " HRGDH- (KLAKLAK)2") complete to be cyclized with Cyclometalated iridium coordination compound
Afterwards, dotted line in the structural representation of part, figure is cyclized to represent terminal carboxyl amidation or be connected with subsequent polypeptides sequence；
Fig. 4 is Cyclometalated iridium coordination compound, cyclized polypeptide Ir-HRGDH, Ir-HRGDH- in the embodiment of the present invention 1,2
(KLAKLAK)2Fluorescence emission spectrogram of compound (ex before and after coordination cyclisation：328nm)；
Fig. 5 is Cyclometalated iridium coordination compound and polypeptide A c-HRGDH-NH in the embodiment of the present invention 22Complete complexation reaction
The mass spectrum of product (Ir-HRGDH) afterwards；
Fig. 6 is Cyclometalated iridium coordination compound in the embodiment of the present invention 2, polypeptide A c-HRGDH-NH2And the two reaction production
The UPLC of thing (Ir-HRGDH)；
Fig. 7 A- Fig. 7 C be in the embodiment of the present invention 2 polypeptide RGD in cyclisation after in different cell lines (A549, MCF-7)
Selectivity enters the laser confocal imaging figure of born of the same parents, wherein, it can see by Fig. 7 A and Fig. 7 B, the RGD (Ir-HRGDH) after cyclisation
With notable differences of the linear RGD (RGDHH-Ir) on born of the same parents' ability is entered；
Fig. 8 is Ac-HRGDH- (KLAKLAK) in the embodiment of the present invention 32It is anti-that coordination is completed with Cyclometalated iridium coordination compound
Cytotoxicity before and after answering (before and after being cyclized) is compared with hemolytic change；
Fig. 9 is to obtain cyclized polypeptide Ir-HRGDH- (KLAKLAK) in the embodiment of the present invention 32(it may be also referred to simply as Ir-
HRGDH-KLA) the quick co-focusing imaging collection of illustrative plates for inducing Apoptosis overall process.
Embodiment
As it was previously stated, in view of deficiency of the prior art, idea of the invention is that providing a kind of new based on metal
The polypeptide fluorescence cyclization method of complex of iridium, it is mainly based upon the complex of iridium that a class is combined generation fluorophor with histidine
And realize.
Further say, should be based on many of metal iridium complex as the more preferred embodiment of the present invention
Peptide fluorescence cyclization method system passes through 2 histidines on the non-blooming Cyclometalated iridium coordination compound of a class itself and polypeptide peptide chain
Residue is combined, and is completed polypeptide cyclisation, is formed the fluorophor that can be excited while optimization polypeptide performance, be able to labeling polypeptide point
Son, and for analyzing detection and fluorescence imaging.
The process of polypeptide cyclisation is simple, quick, efficient in the present invention, and reaction condition is gentle, and the polypeptide nature after cyclisation is steady
It is fixed, without obvious bio-toxicity, further complicated modification and assembling process are can be applied to, and be easy to show with green fluorescence
Track, has broad application prospects in fields such as life sciences, for example, its application may include but be not limited to biomolecule
Detection, cell dyeing, the assembling of animal tissue's mark and polypeptide, mark and the experiment such as spike.
Wherein, Cyclometalated iridium coordination compound is preferred to use by two identical C^N bidentate ligands and two solvents point
The Cyclometalated iridium coordination compound of sub- ligand complex, its chemical molecular formula can be (LC^N)2Ir(Lsolv)2M-, its structural formula can
Refering to foregoing description.
Further, foregoing C^N bidentate ligands, the type of solvent molecule also see above, and here is omitted.
In an exemplary embodiments, the structural formula of foregoing Cyclometalated iridium coordination compound can be：
Wherein ppy is 2-pyridine (2- phenylpyridines), and OTf- is Trifluoromethanesulfonate (trifluoros
Pyrovinic acid root), LsolvTo be easy to the solvent molecule departed from from central metal, such as water or acetonitrile.
Foregoing OTf-Can also hexafluoro-phosphate radical (PF6 -), tetrafluoroborate (BF4 -) etc. replace.
Obvious, to realize that aforementioned coordinative reacts, then it must at least contain two histidines in the polypeptide peptide chain, wherein
Two histidines to carry out complexation reaction with Cyclometalated iridium coordination compound should be distributed in two differences on polypeptide peptide chain
Select location.For linear peptide, two histidines should be distributed in the difference of polypeptide peptide chain along its length
At position.
In another example, the process of foregoing Cyclometalated iridium coordination compound and polypeptides reactive refers to Fig. 1, and wherein H is histidine
Residue, X are to be interval in arbitrary amino acid between two histidine residues, in addition to histidine, and its quantity may be greater than
Or the positive integer equal to 0, preferably 0-5, and be distributed in the amino acid classes of spacer segment and do not limit that (certainly, histidine is removed
Outside).
Further, inventor has found, by steric interference, in polypeptide peptide chain two selected histidine residues it
Between the amino acid quantity that is spaced when being 2-3, cyclisation effect (including reaction rate, generation fluorescence intensity, product stability etc.)
Most preferably.
Postscript, the N-terminal of foregoing polypeptides peptide chain should be free of histidine residues or contain protected histidine residues.More specifically
Say, in the method, Cyclometalated iridium coordination compound carry out complexation reaction when, 2 coordinating groups need to be provided simultaneously, for example
Imidazoles, amino etc..Therefore a free imidazole group, or polypeptide must be provided respectively by two in polypeptide peptide chain histidine
The single histidine of N-terminal provides imidazole group and terminal amino group simultaneously, can complete the complexation reaction.Therefore, to avoid depositing
Be polypeptide peptide chain N-terminal and retain the histidine residues of amino while providing imidazole group and terminal amino group is participated in and ring gold
Belong to the ligand complex reaction of iridium complex compound, the histidine amino of the N-terminal should be protected (such as acetylation).
And if a histidine is located at peptide C end or inside, it can not individually complete aforementioned coordinative reaction, also can not
Obvious green fluorescence is quickly produced, polypeptide cyclisation can not be more realized.
Technical scheme is further described below in conjunction with some preferred embodiments.
Embodiment 1：Pattern polypeptide A c-HAnH-NH2(n=0,1,2,3,4,5) cyclisation effect compares
By [(ppy)2Ir(H2O)2] (OTf) and each one histidine of band of head and the tail pattern polypeptide A c-HAnH (n=0,1,2,
3,4,5) (foregoing A represents alanine) is reacted, [(ppy)2Ir(H2O)2] mol ratio of reaction density of (OTf) and polypeptide is 1：
1, reaction is carried out in PBS (pH7.4), and 37 DEG C of concussions are stayed overnight for 2 hours or 4 DEG C, so that the polypeptide after being cyclized.
Under ultraviolet light (365nm) irradiation, the polypeptide after cyclisation can produce green fluorescence.Head and the tail histidine midfeather
Amino acid quantity is different, the speed of complexation reaction, generates the intensity and the reducing condition (GSH=in analog cell slurry of fluorescence
Stability under 5mM) is slightly different.Spacer amino acids quantity becomes between (referring to Fig. 2A-Fig. 2 D) two histidines of com-parison and analysis
Change and find, spacer amino acids quantity is spaced the cyclisation peptide of 2 or 3 amino acid in reaction rate, produced glimmering when between 0-5
The cyclized polypeptide that 0,1,4,5 amino acid is all relatively spaced in terms of luminous intensity and reducing condition stability inferior has a clear superiority.Wherein,
Work as n=0, when 1, spacer amino acids are very few, cause two histidine intervals short, flexible not enough, reaction is influenceed by steric hindrance；
Work as n=4,5, then spacer amino acids excessive, two histidine hypertelorisms, the progress of the reaction equally limited again.When two groups
It is optimal selection when being spaced 2-3 amino acid between propylhomoserin.
Embodiment 2：Polypeptide A c-HRGDH-NH2Cyclization
By [(ppy)2Ir(H2O)2] (OTf) and each one histidine of band of head and the tail polypeptide A c-HRGDH-NH2Reaction (is referred to
Fig. 3), polypeptide cyclisation process and reaction condition are substantially the same manner as Example 1, and difference is that the polypeptide in the present embodiment is
Ac-HRGDH-NH2.Fig. 4 shows iridium complex compound (Ir is abbreviated as in figure) and polypeptide A c-HRGDH-NH2(it is abbreviated as in figure
HRGDH) fluorescence emission spectrum (the ex before and after coordination cyclisation：328nm).Also, referring to Fig. 5, reaction product is entered with mass spectrum
Row analysis, gained test number (1162.4) is consistent with estimating reaction product (Ir-HRGDH) molecular weight (1163).
Shown again referring to Fig. 6, being tested through UPLC (ultra performance liquid chromatography), after reaction, originally belong to ring metal
The characteristic peak of iridium complex compound (Ir is abbreviated as in figure) and polypeptide (Ac-HRGDH is abbreviated as in figure) is wholly absent, and generates one
Individual new peak (Ir-HRGDH), complete degree of reaction is more than 95%.
Fig. 7 A-7C are referred to again, it can be seen that complete the cyclisation of foregoing rgd peptide sequence using the method for the present embodiment
Afterwards, embodied and entered born of the same parents' ability and thin in RGD identifications positive (A549)/negative (MCF-7) compared with what its linear condition substantially optimized
The selectivity of intercellular.This is consistent with the optimization function that other traditional cyclisation modes of document report are produced to RGD.Also, this reality
The MTT testing results for applying polypeptide after being cyclized in example show to complete the polypeptide cells toxicity that cyclisation is caused by such cyclization method
It is low, LC50More than 200 μM, its result is referred to shown in table 1.
Embodiment 3：Polypeptide A c-HRGDH- (KLAKLAK)2Cyclisation and cytotoxicity optimize
In the present embodiment, polypeptide cyclisation part-structure (also referring to Fig. 3) and cyclization process and reaction condition and embodiment 2
Essentially identical, difference is, the polypeptide in the present embodiment is connected to cytotoxic polypeptide outside cyclisation part
(KLAKLAK)2。(KLAKLAK)2Itself is difficult because entering born of the same parents in the case of monomer, it is impossible to enters cell and induces Apoptosis.And lead to
Cross and be connected with Ir-HRGDH, promote its adhesion that the cell surface being positive is recognized with RGD, effectively help it to enter born of the same parents simultaneously
Play toxicity.
Ac-HRGDH-(KLAKLAK)2The cytotoxicity before and after coordination cyclization is completed with Cyclometalated iridium coordination compound
Compared as shown in Figure 8 with hemolytic change.Completed with Cyclometalated iridium coordination compound after coordination cyclization, polypeptide is embodied
The raising that 40 times of toxicity, and haemolysis property stills remain in relatively low level, surface its there is splendid vivo applications prospect.
Related fluorescence emission spectrum result in the present embodiment is as shown in Fig. 2 cytotoxicity MTT testing results such as table 1 below
It is shown.
The cytotoxicity LC of involved polypeptide and cyclized polypeptide in the embodiment 1-3 of table 150Data.
Simultaneously as the present invention is also green fluorescence on cyclized polypeptide mark while polypeptide cyclisation is completed, make this reality
Apply the polypeptide Ir-HRGDH- (KLAKLAK) after being cyclized in example2The process of Apoptosis is triggered to be able to characterize and (refer to Fig. 9).
The present invention is sent out using 2 histidines of itself non-blooming Cyclometalated iridium coordination compound and select location in peptide chain
Raw complexation reaction, completes polypeptide cyclisation, while forming the fluorophor that can be excited, peptide molecule is labeled fluorescence, can use
In analysis detection and fluorescence imaging etc..
In a word, present invention process is simple and quick, and reaction condition is gentle, and reaction efficiency is high, and product property is stable, without substantially raw
Thing toxicity, modifies or assembles beneficial to further complicated design, with wide biochemistry and medical application prospect.
Be the one or more of preferred embodiment it is pointed out that disclosed, every local change or
Modify and come from the technological thought of the present invention and be that the people for being familiar with this technology is easy to what is deduced, the special of the present invention is not departed from all
Economic rights scope.
Claims (10)
1. a kind of polypeptide fluorescence cyclization method based on metal iridium complex, it is characterised in that including：
Cyclometalated iridium coordination compound is provided, and makes the Cyclometalated iridium coordination compound and the difference being distributed in polypeptide peptide chain
Complexation reaction occurs for two histidine residues at position, realizes polypeptide cyclisation and generation can be by the green glimmering of ultraviolet excitation
Light；
The chemical molecular formula of the Cyclometalated iridium coordination compound is (LC^N)2Ir(Lsolv)2M, its structural formula is as follows：
Wherein, LC^NFor C^N bidentate ligands, LsolvFor solvent molecule, M-Including trifluoromethane sulfonic acid root, hexafluoro-phosphate radical or tetrafluoro
Borate.
2. the polypeptide fluorescence cyclization method based on metal iridium complex according to claim 1, it is characterised in that：The C^N
Bidentate ligand include 2- phenylpyridines, 1- phenyl isoquinolin quinolines, 2- phenyl benzothiophenes, benzoquinoline, 4- methylphenylpyridiniums, 4,
6- difluorophenyl pyridinatos, 2- (benzothiophene) pyridine, 2- phenylchinolines or 2,5- diphenyloxazole.
3. the polypeptide fluorescence cyclization method based on metal iridium complex according to claim 1, it is characterised in that：The solvent
Molecule includes water, acetonitrile, DMF or DMSO.
4. the polypeptide fluorescence cyclization method based on metal iridium complex according to claim 1, it is characterised in that：The coordination
Reaction is carried out in liquid phase environment, and the liquid phase environment is mainly made up of buffer solution system, and the buffer solution system includes phosphorus
Acid buffer, Tris buffer solutions, Bis-Tris buffer solutions, MES buffer solutions or HEPES buffer solution.
5. the polypeptide fluorescence cyclization method based on metal iridium complex according to claim 1, it is characterised in that：Described many
In peptide peptide chain, interval is more than 0 but is less than between two histidines of complexation reaction occur with the Cyclometalated iridium coordination compound
Or any amino acid equal to 5 in addition to histidine.
6. the polypeptide fluorescence cyclization method based on metal iridium complex according to claim 1, it is characterised in that：The polypeptide
Including HRGDH peptides or HRGDH- (KLAKLAK)2Peptide.
7. a kind of cyclized polypeptide, it is characterised in that comprising mainly by Cyclometalated iridium coordination compound with being distributed in polypeptide peptide chain
Two histidine residues at diverse location pass through cyclized structure formed by complexation reaction；
Wherein, the chemical molecular formula of the Cyclometalated iridium coordination compound is (LC^N)2Ir(Lsolv)2M, structural formula is as follows：
Wherein, LC^NFor C^N bidentate ligands, LsolvFor solvent molecule, M-Including trifluoromethane sulfonic acid root, hexafluoro-phosphate radical or tetrafluoro
Borate.
8. cyclized polypeptide according to claim 7, it is characterised in that：LC^NIncluding 2- phenylpyridines, 1- phenyl isoquinolin quinolines,
2- phenyl benzothiophenes, benzoquinoline, 4- methylphenylpyridiniums, 4,6- difluorophenyl pyridinatos, 2- (benzothiophene) pyridine, 2- benzene
Base quinoline or 2,5- diphenyloxazoles.
9. cyclized polypeptide according to claim 7, it is characterised in that：LsolvFor solvent molecule, including water, acetonitrile, DMF or
DMSO。
10. cyclized polypeptide according to claim 7, it is characterised in that in the polypeptide peptide chain, with the Cyclometalated iridium
Complex occurs to be spaced more than 0 less than or equal to 5 in addition to histidine between two histidines of complexation reaction
Any amino acid.
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| CN2013100776342 | 2013-03-12 | ||
| CN2013100776342A CN103204899A (en) | 2013-03-12 | 2013-03-12 | Polypeptide cyclizing method based on combination of histidine and metal iridium complex |
| CN201410079377.0A CN104045683A (en) | 2013-03-12 | 2014-03-06 | Iridium complex-based polypeptide fluorescent cyclisation method and cyclized polypeptide |
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| CN201410079377.0A Pending CN104045683A (en) | 2013-03-12 | 2014-03-06 | Iridium complex-based polypeptide fluorescent cyclisation method and cyclized polypeptide |
| CN201710308188.XA Active CN107129519B (en) | 2013-03-12 | 2014-03-06 | Polypeptide fluorescence cyclization method based on metal iridium complex and cyclized polypeptide |
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| CN108395474A (en) * | 2018-01-30 | 2018-08-14 | 武汉大学 | A kind of method of visible light-inducing pyrazoles coupling phenylalanine class compound |
| CN114617974A (en) * | 2020-12-10 | 2022-06-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Polypeptide albumin nanoparticle and preparation method and application thereof |
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| CN103204899A (en) * | 2013-03-12 | 2013-07-17 | 中国科学院苏州纳米技术与纳米仿生研究所 | Polypeptide cyclizing method based on combination of histidine and metal iridium complex |
| CN103665096B (en) * | 2013-04-19 | 2016-09-28 | 中国科学院苏州纳米技术与纳米仿生研究所 | A kind of method of formation stable nail peptide based on complex of iridium |
| CN104974227B (en) * | 2014-04-04 | 2019-04-23 | 中国科学院苏州纳米技术与纳米仿生研究所 | The amphiphilic film targeting alpha-helix polypeptide of cation and its application |
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| CN101750486A (en) * | 2008-12-19 | 2010-06-23 | 苏州纳凯科技有限公司 | Method for labeling antibody by fluorophore generated by combination of iridium coordination compound and histidine |
| CN103204899A (en) * | 2013-03-12 | 2013-07-17 | 中国科学院苏州纳米技术与纳米仿生研究所 | Polypeptide cyclizing method based on combination of histidine and metal iridium complex |
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| CN103204899A (en) * | 2013-03-12 | 2013-07-17 | 中国科学院苏州纳米技术与纳米仿生研究所 | Polypeptide cyclizing method based on combination of histidine and metal iridium complex |
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| CN108395474A (en) * | 2018-01-30 | 2018-08-14 | 武汉大学 | A kind of method of visible light-inducing pyrazoles coupling phenylalanine class compound |
| CN108395474B (en) * | 2018-01-30 | 2021-02-19 | 武汉大学 | Method for coupling pyrazole with phenylalanine compound through visible light induction |
| CN114617974A (en) * | 2020-12-10 | 2022-06-14 | 中国科学院苏州纳米技术与纳米仿生研究所 | Polypeptide albumin nanoparticle and preparation method and application thereof |
| CN114617974B (en) * | 2020-12-10 | 2023-10-03 | 中国科学院苏州纳米技术与纳米仿生研究所 | Polypeptide albumin nanoparticle and preparation method and application thereof |
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| CN103204899A (en) | 2013-07-17 |
| CN107129519B (en) | 2020-12-22 |
| CN104045683A (en) | 2014-09-17 |
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