Coupling of thiol-containing peptides with fluorinated porphyrins for use as biological probes

Abstract

This invention provides a method for preparing a class of β-peptide-based octafluoroporphyrin metal complexes and their application as biological probes in near-infrared II fluorescence imaging. After β-coupling, the metal complexes can recognize α-peptides. v β3 integrin overexpression in tumor target cells enables specific near-infrared II fluorescence imaging of tumor cells and tissues, providing deeper tissue penetration and higher spatial resolution, and has great application prospects in the field of tumor fluorescence visualization diagnosis and treatment.

Description

Technical Field

[0001] This invention belongs to the field of organic synthesis technology, specifically relating to an octafluoroporphyrin metal complex containing a β-peptide group, and particularly to the application of the octafluoroporphyrin metal complex containing a β-peptide group as a biological probe for near-infrared fluorescence imaging. Background Technology

[0002] Photodynamic therapy utilizes specific photosensitizers to selectively accumulate in tumor tissue, thereby causing targeted removal of tumor tissue through photodynamic killing without affecting normal tissue. Porphyrins, as an important class of photosensitizers, exhibit a special affinity for certain tissues after intravenous injection, selectively remaining in malignant tissues at concentrations far higher than in normal tissues. Specific fluorescence excited by irradiation with specific wavelengths can be used to define the tumor outline, and then irradiation with therapeutic light waves generates a photodynamic effect, killing the tumor.

[0003] Porphyrins are compounds with a large conjugated ring structure found in living organisms. Their basic skeleton is a cyclic conjugated macromolecule, namely porphyrin, formed by four pyrrole rings linked by methylene groups, and it is aromatic. When some or all of the hydrogen atoms on the large conjugated ring of porphyrin are replaced by other groups, porphyrin derivatives are obtained. The four nitrogen atoms at the center can combine with metal ions to form very stable organic complexes, namely metalloporphyrins.

[0004] Porphyrin compounds exhibit strong fluorescence emission in the 600-700 nm range or longer, with a long fluorescence lifetime. Furthermore, their spectral decay characteristics differ from the autofluorescence of biological tissues. These spectral features enable porphyrin compounds to eliminate fluorescence interference from the background surrounding tumor tissue when used to detect tumors.

[0005] In addition, near-infrared light has a strong penetrating ability to biological tissues and causes minimal thermal damage to living cells. This requires that the fluorescence emission wavelength of the photosensitizer be within the therapeutic window (650-1700nm). Among various optical diagnostic methods, near-infrared II (NIR-II, 1000-1700nm) fluorescence imaging has attracted widespread attention due to its advantages such as deeper tissue penetration and higher spatial resolution compared to traditional near-infrared I (650-900nm) fluorescence imaging.

[0006] In molecular design, the substituents at the β and meso positions of porphyrins are modulated, thereby controlling the electronic properties and isomerism of the substituents to achieve a Q-band redshift, allowing the fluorescence emission wavelength of porphyrin compounds to enter the therapeutic window. On the other hand, to improve tumor specificity, the phenyl substituents in porphyrins are bound to known tumor-targeting molecules such as carbohydrates, cholesterol, drug molecules, peptides, and antibodies, enabling tumor detection and treatment. Summary of the Invention

[0007] To address the aforementioned problems, this invention provides an octafluoroporphyrin metal complex containing a β-peptide group. Through molecular design, this compound yields a class of biological probes capable of detecting tumor cells or tissues at near-infrared II fluorescence emission wavelengths, enabling in vivo fluorescence imaging of tumors. It possesses deeper tissue penetration depth and higher spatial resolution, significantly improving the accuracy of fluorescence imaging, thus completing this invention.

[0008] The first aspect of this invention aims to provide the use of an octafluoroporphyrin metal complex containing a β-peptide group as a biological probe. The biological probe is a probe for detecting enzyme expression, such as for recognizing α-peptides. v Biological probes for β3 integrin overexpression.

[0009] A second aspect of the present invention provides an octafluoroporphyrin metal complex containing a β-peptide group. The compound has the structure shown in formula (1):

[0010]

[0011] in,

[0012] M is a metallic element or a metallic element with ligands.

[0013] R are each independently selected from substituted phenyl groups, such as nitrogen-containing heterocyclic substituted alkylphenyl groups, tertiary amine substituted alkylphenyl groups, tertiary phosphine substituted alkylphenyl groups, thioether substituted alkylphenyl groups, phenyl or ether substituted phenyl groups containing sulfonic acid groups and halogen groups. The alkyl group is a C1-C5 alkyl group, preferably a C1-C3 alkyl group. R are the same as or different from each other, preferably the same.

[0014] G is a peptide group containing a thioether bond.

[0015] In one embodiment of the present invention, the peptide group further comprises a protein marker molecule.

[0016] The third aspect of this invention aims to provide a method for preparing octafluoroporphyrin metal complexes containing a β-peptide group. The method utilizes a meso-substituted phenyl-β-octafluoroporphyrin metal complex and a peptide via a thiol nucleophilic substitution coupling reaction. The method specifically includes the following steps:

[0017] Step 1: Preparation of meso-substituted phenyl-β-octafluoroporphyrin metal complexes;

[0018] Step 2: Add the meso-substituted phenyl-β-octafluoroporphyrin metal complex and peptide to a biological buffer solution to obtain a reaction solution;

[0019] Step 3: Post-process the reaction solution to obtain an octafluoroporphyrin metal complex containing a β-peptide group.

[0020] The octafluoroporphyrin metal complex containing a β-peptide group provided by this invention has the following beneficial effects as a biological probe:

[0021] (1) In this invention, through molecular design, nitrogen-containing heterocyclic substituent alkylphenyl, tertiary amine substituent alkylphenyl, tertiary phosphine substituent alkylphenyl, thioether substituent alkylphenyl, phenyl or ether substituent alkylphenyl containing sulfonic acid group and halo group are inserted at the meso position of the porphyrin ring, thereby realizing bioorthogonal click chemistry. Furthermore, the different charges of different groups will affect the water solubility and coupling activity of the complex to varying degrees, thereby achieving broad biological functionality.

[0022] (2) The octafluoroporphyrin metal complex containing a β-peptide group provided by this invention is obtained by coupling the porphyrin β-carbon-fluorine bond with a peptide chain containing a cysteine ​​residue or other biofunctional molecules containing a thiol group. This coupling can enhance the biotargeting of the octafluoroporphyrin metal complex. In particular, it can enhance the biotargeting of octafluoroytterbium porphyrin that emits light in the near-infrared II region (1000-1700 nm), thereby enhancing the spatiotemporal resolution of in vivo imaging.

[0023] (3) The octafluoroporphyrin metal complex containing a β-peptide group provided by this invention has biologically specific recognition capabilities. Through molecular design, an octafluoroporphyrin metal complex containing a target peptide chain at the β-position can be constructed, which can be used as a biological probe to recognize and detect intracellular biological processes, and as a fluorescent probe to achieve targeted fluorescence imaging of cells and tumors. Attached Figure Description

[0024] Figure 1 The normalized fluorescence emission spectra of 7a to 7f obtained in Example 4 of the present invention are shown.

[0025] Figure 2 The image shows a fluorescence imaging diagram of HepG2 liver cancer cells or LO2 sections of hepatocytes after treatment for 6a or 7d in Example 5 of the present invention.

[0026] Figure 3 The image shows HepG2 liver cancer tissue cells and healthy liver cells LO2 after 6a or 7d treatment in Example 5 of the present invention, showing Yb. 3+ content;

[0027] Figure 4 This illustrates the Yb content in HepG2 liver cancer cells from the azide group, iRGD group, and control group in Example 6 of the present invention. 3+ content;

[0028] Figure 5The image shows fluorescence imaging of HeLa cells and HepG2 liver cancer cells after 7 days of treatment in Example 7 of the present invention;

[0029] Figure 6 The image shows fluorescence imaging of mice injected with an aqueous solution for 7 days in Example 8 of the present invention, as well as in vitro fluorescence imaging of their heart, liver, spleen, lung, kidney and tumor. Detailed Implementation

[0030] The present invention will now be described in detail through specific embodiments, and the features and advantages of the present invention will become clearer and more explicit with these descriptions.

[0031] The octafluoroporphyrin metal complex containing a β-peptide group in this invention utilizes the thiol group to bind with various amino acid residues and peptides, enabling porphyrin to specifically select and recognize tumors or biological tissues at different locations, accurately perform fluorescence imaging of tumors or biological tissues, and its fluorescence emission wavelength can reach the near-infrared II region (NIR-II, 1000-1700nm), giving its fluorescence imaging a deeper tissue penetration depth and higher spatial resolution, significantly improving the accuracy of photosensitizer imaging.

[0032] The first aspect of this invention provides the use of an octafluoroporphyrin metal complex containing a β-peptide group as a biological probe.

[0033] The peptide group is coupled to a β-octafluoroporphyrin metal complex via a thiol group. The peptide chain (peptide group) used for coupling contains a thiol group, for example, the peptide chain (peptide group) contains a cysteine ​​residue or is modified with other groups containing a thiol group. The other groups containing a thiol group are preferably selected from mercaptoalkylamine or mercaptoalkylcarbonyl groups. Preferably, the peptide group is coupled to the β-octafluoroporphyrin metal complex at the β-position.

[0034] In one embodiment of the present invention, the peptide group further comprises a protein marker molecule selected from CY series cyanine dyes, such as CY3, CY5, CY5.5, CY7, CY7.5, fluorescein isothiocyanate (FITC), boron dipyrrole (Bodipy) series dyes, such as rhodamine B and its derivatives (such as tetramethylrhodamine isothiocyanate TRITC), coumarin and two-photon coumarin series dyes, indocyanine green (ICG) dyes, such as indocyanine green and indocyanine green carboxylic acid, preferably fluorescein isothiocyanate (FITC), rhodamine B, or indocyanine green dyes, more preferably fluorescein isothiocyanate (FITC).

[0035] In one embodiment of the present invention, the biological probe is a biological probe for detecting enzyme expression, such as one used to identify α-protein. v Biological probes for β3 integrin.

[0036] In another embodiment of the present invention, the octafluoroporphyrin metal complex bioprobe containing a β-peptide group is used to prepare biological agents for the diagnosis and treatment of tumor tissues, such as photosensitizers for photodynamic therapy or imaging agents for near-infrared cells or tissues of tumors.

[0037] A second aspect of the present invention provides an octafluoroporphyrin metal complex containing a β-peptide group. The compound has the structure shown in formula (1):

[0038]

[0039] in,

[0040] M is a metallic element or a metallic element with a ligand, wherein the metallic element is a transition metal element, preferably one of zinc, manganese, iron, palladium, platinum, ytterbium, gadolinium, lutetium, erbium, and neodymium, more preferably one of manganese, ytterbium, gadolinium, lutetium, erbium, and neodymium, such as ytterbium. The ligand is... The ligand, acetylacetone ligand, pyrazolborane ligand, or phosphooxy ligand is preferred. Ligands.

[0041] R are each independently selected from substituted phenyl groups, such as nitrogen-containing heterocyclic substituted alkylphenyl groups, tertiary amine substituted alkylphenyl groups, tertiary phosphine substituted alkylphenyl groups, thioether substituted alkylphenyl groups, phenyl or ether substituted phenyl groups containing sulfonic acid groups and halogen groups. The alkyl group is a C1-C5 alkyl group, preferably a C1-C3 alkyl group. R are the same as or different from each other, preferably the same.

[0042] Preferably,

[0043] The nitrogen-containing heterocyclic substituent is selected from pyrrole substituents, pyridine substituents, pyrimidine substituents, quinoline substituents, imidazole substituents, or pyrazole substituents, preferably pyrrole, pyridinyl, pyrimidinyl, quinolineyl, imidazoleyl, or pyrazolyl, and more preferably pyridinyl.

[0044] The tertiary amine substituents are selected from alkyl tertiary amine, alcohol tertiary amine, alkoxy tertiary amine, alkynyl tertiary amine, azidoalkyl tertiary amine, and preferably trimethylamine, hydroxyethyl-dimethylamine, 2-[2-(dimethylamino)ethoxy]ethanol, dimethylpropynylamine, or dimethylazidoethylamine.

[0045] The tertiary phosphine substituent is selected from alkylphosphinyl or phenylphosphinyl, preferably trimethylphosphinyl or triphenylphosphinyl.

[0046] The thioether substituents are selected from alkyl thioethers, preferably dimethyl thio.

[0047] More preferably, R is independently selected from group 1 to group 11, and may be the same as or different from each other, preferably the same:

[0048]

[0049] (where Ph is phenyl). At this point, the octafluoroporphyrin metal complexes containing the β-peptide group are respectively β-peptide-S-heptafluoroporphyrin metal complex I to β-peptide-S-heptafluoroporphyrin metal complex XI.

[0050] In this invention, groups 1 to 11 are preferably used to substitute the meso position of the porphyrin ring. The alkynyl group in group 5 and the azide group in group 6 can achieve bioorthogonal click chemistry. Groups 1-9 are positively charged, group 10 is electroneutrally neutral, and group 11 is electronegative. Different charges will affect the water solubility, coupling activity, and biofunctionality of the complex to varying degrees.

[0051] G is a peptide group containing a thioether bond. This includes peptide groups containing cysteine ​​residues, thioether-alkylamine groups, or thioether-alkylcarbonyl groups, preferably polypeptide groups containing mercaptopropionic acid groups, cysteine ​​residues, or mercaptoacetic acid groups. G is coupled to a β-octafluoroporphyrin-type metal complex at the β-position via a thioether bond. The peptide group is a polypeptide group that acts as a substituent.

[0052] The peptide contains 2-100 amino acid residues, preferably 3-50, more preferably 3-10, such as glutathione, FCPF-, c(RGDFC)-, c(RGDFK(MPa))- (MPa is mercaptopropionic acid), DEVDGC- (where the letters are general abbreviations for amino acids).

[0053] Preferably, the octafluoroporphyrin metal complex containing a β-peptide group is selected from:

[0054] β-GSH-S-heptafluoroporphyrin metal complexes I to β-GSH-S-heptafluoroporphyrin metal complexes XI, where GSH is a glutathione group.

[0055] β-FCPF-S-heptafluoroporphyrin metal complex I to β-FCPF-S-heptafluoroporphyrin metal complex XI,

[0056] β-c(RGDFC)-S-heptafluoroporphyrin metal complex I to β-c(RGDFC)-S-heptafluoroporphyrin metal complex XI,

[0057] β-c(RGDFK(MPa))-OOC-CH2-CH2-S-heptafluoroporphyrin metal complex I ~ β-c(RGDFK(MPa))-OOC-CH2-CH2-S-heptafluoroporphyrin metal complex XI,

[0058] β-DEVDGC-S-heptafluoroporphyrin metal complex I to β-DEVDGC-S-heptafluoroporphyrin metal complex XI.

[0059] In one embodiment of the present invention, the peptide group further comprises a protein labeling molecule. The protein labeling molecule is selected from CY series cyanine dyes, such as CY3, CY5, CY5.5, CY7, CY7.5, fluorescein isothiocyanate (FITC), boron dipyrrole (Bodipy) series dyes, such as rhodamine B and its derivatives (e.g., TRITC), coumarin and two-photon coumarin series dyes, indocyanine green (ICG) dyes, such as indocyanine green and indocyanine green carboxylic acid, biotin (vitamin H), preferably fluorescein isothiocyanate (FITC), rhodamine B, indocyanine green dyes, or biotin (vitamin H), more preferably fluorescein isothiocyanate (FITC) or biotin (vitamin H). Examples include β-(Biotin-GSH)-S-heptafluoroporphyrin metal complex I to β-(Biotin-GSH)-S-heptafluoroporphyrin metal complex XI, and β-(DEVDGC-FITC)-S-heptafluoroporphyrin metal complex I to β-(DEVDGC-FITC)-S-heptafluoroporphyrin metal complex XI.

[0060] The octafluoroporphyrin metal complex containing a β-peptide group is first obtained by coupling a meso-substituted β-octafluoroporphyrin metal complex with a thiol biomolecule via thiol nucleophilic substitution.

[0061] Preferably, the octafluoroporphyrin metal complex containing the β-peptide group is selected from 7a to 7g:

[0062]

[0063]

[0064]

[0065] Among them, Yb coordinates with the oxygen in the three "P=O" bonds in L.

[0066] The octafluoroporphyrin metal complex containing a β-peptide group provided by this invention couples a peptide chain containing a cysteine ​​residue or other molecules containing a thiol group to an octafluoroporphyrin, achieving biologically transparent near-infrared II (1000–1700 nm) fluorescence imaging, enhancing detection depth, and more importantly, endowing the metalloporphyrin with biologically specific recognition capabilities. Through molecular design, an octafluoroporphyrin metal complex containing a β-peptide group and a target peptide chain is constructed, which can be used as a biological probe to recognize and detect intracellular biological processes. It can also be used as an anticancer photosensitizer to detect tumor cells.

[0067] A third aspect of this invention provides a method for preparing octafluoroporphyrin metal complexes containing a β-peptide group. The method utilizes a meso-substituted phenyl-β-octafluoroporphyrin metal complex and a polypeptide via a thiol nucleophilic substitution reaction.

[0068] The method specifically includes the following steps:

[0069] Step 1: Prepare meso-substituted phenyl-β-octafluoroporphyrin metal complex.

[0070] Step 1 specifically includes the following sub-steps:

[0071] Sub-step a, meso-substituted phenyl-β-octafluoroporphyrin.

[0072] In the meso-substituted phenyl-β-octafluoroporphyrin, the substituted phenyl group is selected from halogenated alkylphenyl groups, phenyl groups containing sulfonic acid groups and halogenated groups, or ether-substituted phenyl groups, preferably chloroalkylphenyl, bromoalkylphenyl, iodoalkylphenyl, group 10 or group 11, more preferably bromoalkylphenyl, group 10 or group 11. The alkyl group is a C1-C5 alkyl group, preferably a C1-C3 alkyl group.

[0073] Preferably, the meso-substituted phenyl-β-octafluoroporphyrin is meso-haloalkylphenyl-β-octafluoroporphyrin, meso-(sulfonyl halophenyl)-β-octafluoroporphyrin, or meso-(ether substituted phenyl)-β-octafluoroporphyrin.

[0074] In one embodiment of the present invention, meso-haloalkylphenyl-β-octafluoroporphyrin is obtained by condensation of 3,4-difluoropyrrole and p-haloalkylbenzaldehyde (or m-haloalkylbenzaldehyde, or or o-haloalkylbenzaldehyde).

[0075] The p-haloalkylbenzaldehyde (or m-haloalkylbenzaldehyde, or o-haloalkylbenzaldehyde) is selected from p-halomethylbenzaldehyde (or m-haloalkylbenzaldehyde, or o-haloalkylbenzaldehyde), preferably p-chloromethylbenzaldehyde (or m-haloalkylbenzaldehyde, or o-haloalkylbenzaldehyde), p-bromomethylbenzaldehyde (or m-haloalkylbenzaldehyde, or o-haloalkylbenzaldehyde), or p-iodomethylbenzaldehyde (or m-haloalkylbenzaldehyde, or o-haloalkylbenzaldehyde), preferably 4-bromomethylbenzaldehyde.

[0076] In another embodiment of the present invention, meso-(sulfonyl halophenyl)-β-octafluoroporphyrin and meso-(ether substituent phenyl)-β-octafluoroporphyrin are obtained by condensation of 3,4-difluoropyrrole with o-dihalobenzaldehyde and m-ether-type benzaldehyde, respectively.

[0077] The molar ratio of the 3,4-difluoropyrrole to the benzaldehyde compound is 1:(0.6-1.6), preferably 1:(0.8-1.4), and more preferably 1:(1-1.2). The benzaldehyde compound is p-haloalkylbenzaldehyde, o-dihalobenzaldehyde, or m-ether benzaldehyde.

[0078] The reaction is carried out in the presence of a catalyst selected from boron trifluoride ethers, preferably boron trifluoride diethyl ether. The volume molar ratio of the catalyst to 3,4-difluoropyrrole is (5-45) μL:2.0 mmol, preferably (10-35) μL:2.0 mmol, and more preferably (15-25) μL:2.0 mmol.

[0079] The reaction is carried out in the presence of a solvent selected from one or more of halogenated hydrocarbon solvents, ether solvents, and ketone solvents, preferably selected from one or more of dichloromethane, trichloromethane, tetrahydrofuran, propylene oxide, and acetone, and more preferably anhydrous dichloromethane. The volume molar ratio of the solvent to 3,4-difluoropyrrole is (300-520) mL:2.0 mmol, preferably (350-470) mL:2.0 mmol, and more preferably (380-420) mL:2.0 mmol.

[0080] After adding the catalyst to the reaction solution containing 3,4-difluoropyrrole, benzaldehyde compounds and solvent, the mixture is stirred for 1-15 hours, preferably 11-13 hours, at a temperature of 15-30°C, preferably 20-25°C.

[0081] Then an oxidant is added, and the reaction is carried out for 1.0-5.5 h, preferably 3.5-4.5 h. The oxidant is selected from benzoquinone compounds, preferably 2,3-dichloro-5,6-dicyano-1,4-benzoquinone. The molar ratio of the oxidant to 3,4-difluoropyrrole is (1.6-2.6):2.0, preferably (1.8-2.4):2.0, and more preferably (2.0-2.2):2.0.

[0082] After the reaction was complete, the reaction solution was transferred to a silica gel or neutral alumina fast column and eluted with anhydrous dichloromethane. The solvent was removed by vacuum distillation, and the residual solids were separated on a silica gel column using a mixed solvent of petroleum ether and dichloromethane (volume ratio 3:1) as the eluent to obtain meso-substituted phenyl-β-octafluoroporphyrin.

[0083] Sub-step b: Prepare meso-substituted phenyl-β-octafluoroporphyrin metal complex.

[0084] The meso-substituted phenyl-β-octafluoroporphyrin metal complex is obtained by complexing meso-substituted phenyl-β-octafluoroporphyrin with a metal compound.

[0085] In one embodiment of the present invention, when the benzaldehyde compound is p-haloalkylbenzaldehyde (or m-haloalkylbenzaldehyde, or o-haloalkylbenzaldehyde), 3,4-difluoropyrrole and p-haloalkylbenzaldehyde (or m-haloalkylbenzaldehyde, or o-haloalkylbenzaldehyde) are condensed to obtain meso-haloalkylphenyl-β-octafluoroporphyrin. After being coordinated with a metal compound, it is then reacted with one of the following compounds: nitrogen-containing heterocyclic compound, tertiary amine compound, tertiary phosphine compound, or thioether compound to prepare a meso-substituted phenyl-β-octafluoroporphyrin metal complex.

[0086] In another embodiment of the present invention, when the benzaldehyde compound is selected from ortho-dihalobenzaldehyde and meta-ether benzaldehyde, the meso-substituted phenyl-β-octafluoroporphyrin metal complex is obtained by complexing meso-(sulfonyl halophenyl)-β-octafluoroporphyrin or meso-(ether substituted phenyl)-β-octafluoroporphyrin with a metal compound, respectively.

[0087] The metal compound is selected from zinc compounds, manganese compounds, iron compounds, palladium compounds, platinum compounds or lanthanide metal compounds, preferably lanthanide metal compounds, more preferably ytterbium compounds, gadolinium compounds, lutetium compounds, erbium compounds or neodymium compounds, such as ytterbium acetylacetone hydrate (Yb(acac)3·3H2O).

[0088] The molar ratio of the meso-substituted phenyl-β-octafluoroporphyrin to the metal compound is 1:(0.3-0.9), preferably 1:(0.4-0.8), and more preferably 1:(0.5-0.7).

[0089] The reaction is carried out in the presence of a solvent selected from one or more of sulfone solvents, aromatic hydrocarbon solvents, and amide solvents, preferably one or more of sulfoxide solvents, halobenzene solvents, N,N-dimethylacetamide, and N,N-dimethylformamide, more preferably one or more of 1,2,4-trichlorobenzene (TCB), dimethyl sulfoxide, or N,N-dimethylformamide (DMF).

[0090] The reaction temperature is 100-270℃, preferably 150-250℃; the reaction time is 15-60 min, preferably 20-40 min.

[0091] After the reaction was completed, the mixture was cooled to room temperature and separated by passing the reaction mixture through a silica gel column. The intermediate product was obtained by using a mixed solvent of DCM (dichloromethane) and methanol (DCM to methanol volume ratio of 5:1) as the eluent.

[0092] Preferably, the intermediate product is combined with The ligand compound was added to the solvent and reacted with stirring at room temperature to obtain meso-haloalkylphenyl-β-octafluoroporphyrin metal complexes, meso-(sulfonylhalophenyl)-β-octafluoroporphyrin metal complexes, or meso-(ether-substituted phenyl)-β-octafluoroporphyrin metal complexes.

[0093] If the ligand is NaL OCD3 (sodium[(cyclopentadienyl)tris(-di(methyl-d3)phosphito)cobaltate](D(deuterium)atom>99%), which was synthesized according to the method described in the literature "Inorg. Chem., 2012, 51, 12436–12443.")

[0094] In one embodiment of the present invention, the meso-substituted phenyl-β-octafluoroporphyrin metal complex is prepared by reacting the meso-haloalkylphenyl-β-octafluoroporphyrin metal complex with one of the following compounds: nitrogen-containing heterocyclic compounds, tertiary amine compounds, tertiary phosphine compounds, and thioether compounds.

[0095] The molar ratio of the nitrogen-containing heterocyclic compound, tertiary amine compound, tertiary phosphine compound, thioether compound, and sulfuric acid to meso-halosubstituted phenyl-β-octafluoroporphyrin is (4-950):1, preferably (4-900):1, and more preferably (4-850):1.

[0096] The reaction temperature and reaction time are determined based on the reaction of the nitrogen-containing heterocyclic compounds, tertiary amine compounds, tertiary phosphine compounds, and thioether compounds with meso-halosubstituted phenyl-β-octafluoroporphyrin. The reaction is preferably carried out at reflux temperature, and after crystallization and precipitation, the product is purified by reversed-phase high-performance liquid chromatography.

[0097] The nitrogen-containing heterocyclic compound is selected from pyrrole compounds, pyridine compounds, pyrimidine compounds, quinoline compounds, imidazole compounds or pyrazole compounds, preferably selected from pyrrole, pyridine, pyrimidinequinoline, imidazole or pyrazole, and more preferably pyrrole.

[0098] The tertiary amine compounds are selected from alkyl tertiary amines, alcohol tertiary amines, alkoxy tertiary amines, alkynyl tertiary amines, and azidoalkyl tertiary amines, preferably trimethylamine, 2-(N,N-dimethylamino)ethanol, 2-[2-(dimethylamino)ethoxy]ethanol, N,N-dimethylpropynylamine, or N,N-dimethylazidoethylamine (DMAZ).

[0099] The tertiary phosphine compounds are selected from alkylphosphine or phenylphosphine, preferably trimethylphosphine or triphenylphosphine.

[0100] The thioether compounds are selected from alkyl thioethers, preferably dimethyl thioethers.

[0101] Step 2: Add the meso-substituted phenyl-β-octafluoroporphyrin metal complex and peptide to a biological buffer solution to obtain a reaction solution.

[0102] The biological buffer is selected from borate buffer, citrate buffer, PB buffer, PBS buffer, Tris-HCl buffer, NH3-NH4Cl buffer, 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES) buffer, preferably PBS buffer, Tris-HCl buffer, Na2CO3-NaHCO3 buffer, and more preferably tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl) solution or Na2CO3-NaHCO3 buffer.

[0103] The molar ratio of the meso-substituted phenyl-β-octafluoroporphyrin metal complex to the peptide is 1:(1-3.2), preferably 1:(1.3-2.8), and more preferably 1:(1.5-2.5).

[0104] The molar volume ratio of the meso-substituted phenyl-β-octafluoroporphyrin metal complex to the biological buffer is 1 mmol:(0.4-1.8) mL, preferably 1 mmol:(0.6-1.5) mL, and more preferably 1 mmol:(0.8-1.2) mL.

[0105] The reaction can be carried out with or without a reducing agent, which is selected from dithiothreitol (DTT), tertiary phosphine such as tris(2-carboxyethyl)phosphine (TCEP), trimethylphosphine, and preferably tris(2-carboxyethyl)phosphine (TCEP). The molar ratio of the meso-substituted phenyl-β-octafluoroporphyrin metal complex to the reducing agent is 1:(2-8), preferably 1:(3-7), and more preferably 1:(4-6).

[0106] The reaction pH is 5.5-12, preferably 6.5-11, and more preferably 7.5-10.0.

[0107] The reaction temperature is 15-37℃, preferably 20-32℃, and more preferably 25-30℃.

[0108] Step 3: Post-process the reaction solution to obtain an octafluoroporphyrin metal complex containing a β-peptide group.

[0109] Mono- and disubstituted products were separated by reversed-phase RP-HPLC using a ReproSil-Pur Basic-C18 column and a mobile phase solvent of 0.01%-0.06% (v / v) trifluoroacetic acid aqueous solution and methanol to obtain octafluoroporphyrin metal complexes containing β-peptide groups.

[0110] The octafluoroporphyrin metal complex containing a β-peptide group provided by this invention can serve as a biological probe to effectively identify overexpressed α-porphyrin. v β3 integrins target cancer cells, thereby enabling specific recognition of tumor tissue. Furthermore, their fluorescence emission wavelength is 950-1100 nm, allowing for fluorescence imaging in the near-infrared II region, providing deeper tissue penetration and higher spatial resolution.

[0111] Example

[0112] Example 1

[0113] 3,4-Difluoropyrrole (206 mg, approx. 2.0 mmol) and 4-bromomethylbenzaldehyde (400 mg, approx. 2.0 mmol) were dissolved in 400 mL of anhydrous dichloromethane (DCM), and the mixture was stirred for 5 minutes. 20 μL (approx. 0.16 mmol) of boron trifluoride diethyl ether (BF3·Et2O) was then added. The mixture was stirred at room temperature for 12 hours. Then, 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) (476 mg, approx. 2.1 mmol) was added, and the reaction mixture was stirred for 4 hours. The reaction mixture was then transferred to a silica gel rapid column and eluted with DCM. The solution obtained by vacuum distillation was separated on a silica gel column using petroleum ether and dichloromethane (volume ratio 3:1) as eluent to give a dark brown 2,3,7,8,12,13,17,18-octafluoro-5,10,15,20-tetra(4-bromomethylphenyl)porphyrin (code name: compound 2, 150 mg, molar yield 66%).

[0114]

[0115] The nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS) data of compound 2 were obtained as follows:

[0116] 1 ¹H NMR (400MHz, deuterated chloroform) δ 8.01 (d, J = 8.0Hz, 8H), 7.75 (d, J = 8.0Hz, 8H), 4.80 (s, 8H), -4.18 (s, 2H).

[0117] 19 F NMR (471 MHz, deuterated chloroform) δ -140.03 (s, 4F), -145.14 (s, 4F).

[0118] HRMS (MALDI-FTICR), m / z [M+H] + [C48H27Br4F8N4] + Calculated value: 1126.8836; Measured value: 1126.8805.

[0119] Example 2

[0120] (1) Compound 2 (115 mg, about 0.1 mmol) obtained in Example 1 and 300 mg of Yb(acac)3·3H2O (ytterbium acetylacetonate hydrate, about 0.64 mmol) were added to a Schlenk tube containing 5 mL of anhydrous 1,2,4-trichlorobenzene (TCB). The mixture was refluxed under nitrogen or argon atmosphere for 30 minutes at 170 °C. After the reaction was completed, the mixture was cooled to room temperature, and the reaction mixture was separated by silica gel column chromatography using a mixed solvent of DCM and methanol (DCM to methanol volume ratio of 5:1) as the eluent to obtain an intermediate product. The obtained intermediate product (which is the Yb complex of compound 2, with acac as an auxiliary ligand) and 50 mg of NaL were added to the solution. OCD3 (Approximately 0.1 mmol) was dissolved in 6 mL of a mixed solvent of chloroform and methanol (volume ratio 5:1). The mixture was stirred at room temperature for 30 minutes, and then the reaction mixture was separated by silica gel column chromatography using a mixed solvent of petroleum ether and DCM (volume ratio 2:1) as eluent to give 6a' as a red solid (108 mg; molar yield 61% based on the addition of compound 2). (Where NaL...) OCD3 The sample was sodium [(cyclopentadienyl)tris(-di(methyl-d3)phosphito)cobaltate] (D(deuterium)atom>99%), synthesized according to the method described in the literature "Inorg. Chem., 2012, 51, 12436–12443."). 6α' nuclear magnetic resonance (NMP) and high-resolution mass spectrometry (HRMS (MALDI-FTICR)) data were obtained.

[0121] 1 ¹H NMR (400 MHz, deuterated chloroform) δ 16.92 (s, 4H), 10.71 (s, 4H), 8.99 (s, 8H), 6.17 (s, 8H), -5.21 (s, 5H).

[0122] 19 F NMR (471 MHz, deuterated chloroform) δ-132.09 (s, 8F).

[0123] HRMS (MALDI-FTICR) [M+H + The m / z of C59H30Br4CoD18F8N4O9P3Yb is calculated to be 1767.9085; the measured value is 1767.9065.

[0124]

[0125] (Oxygen coordination between Yb and the three P=O bonds in L)

[0126] 6a′ (53 mg, approximately 0.03 mmol) was dissolved in 1–2 mL of pyridine (as both solvent and reactant) and refluxed at 80 °C for 2 h. The mixture was cooled to room temperature, and 5 mL of hexane was added to precipitate the crude product. Furthermore, it was purified by RP-HPLC (reversed-phase high-performance liquid chromatography) to obtain the desired product complex 6a as a red solid (39 mg, 73%). NMR and high-resolution mass spectrometry data for 6a were obtained.

[0127] 1 ¹H NMR (400 MHz, deuterated methanol-d⁴) δ 17.40 (s, 4H), 10.99 (s, 4H), 10.59 (d, J = 5.7 Hz, 8H), 9.38 (t, J = 7.9 Hz, 4H), 9.24–9.14 (m, 8H), 9.09 (dd, J = 7.3, 6.7 Hz, 8H), 7.63 (s, 8H), -5.40 (s, 5H).

[0128] 19 F NMR (471 MHz, deuterated methanol-d4) δ -134.11 (s, 8F).

[0129] HRMS(ESI+-FTICR)[M 4+ Calculate C79H49CoD18F8N8O9P3Yb 4+ The calculated m / z value is 441.8486; the measured value is 441.8478.

[0130] (2) 6a′ (53 mg, approximately 0.03 mmol) was dissolved in 1–2 mL of trimethylamine ethanol solution (trimethylamine content 30%) and refluxed at 80 °C for 2 h. The mixture was cooled to room temperature, and 5 mL of hexane was added to precipitate the crude product. Furthermore, it was purified by RP-HPLC (reversed-phase high-performance liquid chromatography) to obtain the desired product complex 6b as a red solid (39 mg, 73%). The NMR and high-resolution mass spectrometry data of 6a were obtained:

[0131]

[0132] (Oxygen coordination between Yb and the three P=O bonds in L)

[0133] 1 ¹H NMR (400MHz, deuterated methanol-d4) 1H NMR (400MHz, Methanol-d4) δ17.66(s,4H),11.12(s,4H),9.19(s,8H),6.32(s,8H),4.52(s,36H),-5.38(s,5H).

[0134] 19 F NMR (471 MHz, deuterated methanol-d4) δ -134.11 (s, 8F).

[0135] HRMS(ESI+-FTICR)[M 4+ Calculate C71H65CoD18F8N8O9P3Yb 4+ The obtained m / z value is 421.8799; the measured value is 421.8790.

[0136] Example 3

[0137] In a light-protected environment at room temperature, the complex 6a prepared in Example 2 and glutathione (GSH) were added to 1 mL of Na₂CO₃-NaHCO₃ buffer, followed by TCEP (tris(2-carboxyethyl)phosphine). The mixture was stirred until the final concentrations of the three compounds were 1 mmol / L, 2 mmol / L, and 5 mmol / L, respectively. The pH was 10.0, and the reaction was carried out for 12 h. After the reaction, the crude mixture was analyzed by HPLC (ReproSil-Pur Basic-C18 column, 5 μm silica gel particle size, 250 × 4.6 mm column length and diameter). The main peaks were then identified by HRMS. The mobile phase solvent was a methanol solution of 0.05% trifluoroacetic acid (v / v). The mono / disubstituted products were separated by reversed-phase RP-HPLC using a ReproSil-Pur Basic-C18 column (5 μm silica gel particle size, 250 × 10 mm column length and diameter). The mobile phase solvent was a methanol solution of 0.05% trifluoroacetic acid by volume, yielding monocoupled product 7a.

[0138] Analysis of the reaction solution after the reaction was completed using high performance liquid chromatography (HPLC) (detection wavelength 395 nm) showed that the molar conversion rate of the reactants was 25% and the selectivity of 7a was 24%.

[0139]

[0140] (Oxygen coordination between Yb and the three P=O bonds in L)

[0141] 7a: HRMS (ESI+-FTICR) [M 4+ Calculate C89H65CoD18F7N11O15P3SYb 4+m / z 513.6180; measured value: 513.6189. [M 4+ Calculate C89H64CoD18F7N11O15P3SYb using -H+] 3+ The measured value is 684.4883: 684.4888. [M] 4+ -H + Calculate C91H65CoD18F10N11O17P3SYb using the TFA calculation. 3+ The measured value was 722.4859; the actual value was 722.4868, and TFA is trifluoroacetic acid.

[0142] Example 4

[0143] Compounds 7b, 7c, 7d, 7e, 7f and 7g were synthesized sequentially according to the method in Example 3. The specific reaction conditions are shown in Table 1. The pH value of the reaction solution was 10.0 and the reaction time was 12 h.

[0144] Table 1:

[0145]

[0146] The peptides in Table 1, including the cyclic peptide RGDFK with a mercaptopropionic acid group (MPa), the FITC-labeled peptide DEVDGC, and the Biotin (vitamin H)-labeled GSH, were customized from Qiangyao Biotechnology.

[0147] The specific results of the 7b-7g structures and HRMS tests are as follows:

[0148]

[0149] (Oxygen coordination between Yb and the three P=O bonds in L)

[0150] 7b: HRMS (ESI+-FTICR) [M4+] Calculation of C105H80CoD18F7N12O14P3SYb 4+ m / z 564.8994; Measured value: 564.8996; [M 4+ -H + Calculate C105H79CoD18F7N12O14P3SYb 3+ The actual value was 752.8634; the measured value was 752.8637.

[0151]

[0152] (Oxygen coordination between Yb and the three P=O bonds in L)

[0153] 7c: HRMS (ESI+-FTICR) [M 4+Calculate C103H82CoD18F7N16O16P3SYb 4+ m / z 581.4038; Measured value: 581.40438. [M 4+ +H + Calculate C103H83CoD18F7N16O16P3SYb 5+ The measured value is 465.3245; the actual value is 465.3255. [M4+-H++TFA] Calculate C105H82CoD18F10N16O18P3SYb 3+ The measured value is 812.8670; the actual value is 812.8666.

[0154]

[0155] (Oxygen coordination between Yb and the three P=O bonds in L)

[0156] 7d: HRMS(ESI+-FTICR)[M 4+ +H + Calculate C109H94CoD18F7N17O17P3SYb 5+ m / z: 487.9413; Measured value: 487.942. [M] 4+ Calculate C109H93CoD18F7N17O17P3SYb 4+ The measured value was 609.6749; the actual value was 609.6762.

[0157]

[0158] (Oxygen coordination between Yb and the three P=O bonds in L)

[0159] 7e: HRMS(ESI+-FTICR)[M4+] Calculation of C104H86CoD18F7N14O23P3SYb 4+ m / z 606.4012; Measured value: 606.4006. [M 4+ -H + Calculate C104H85CoD18F7N14O23P3SYb 3+ 808.1992; Measured value: 808.2017.

[0160]

[0161] (Oxygen coordination between Yb and the three P=O bonds in L)

[0162] 7f: HRMS(ESI+-FTICR)[M 4+Calculate C99H79CoD18F7N13O17P3S2Yb 4+ m / z 570.1374; Measured value: 570.1373. Calculate C99H78CoD18F7N13O17P3S2Yb using [M4+-H+]. 4+ 759.8475; Measured value: 759.8471.

[0163]

[0164] (Oxygen coordination between Yb and the three P=O bonds in L)

[0165] 7g: HRMS(ESI+-FTICR)[M 4+ Calculate C129H106CoD18F7N16O28P3S2Yb 4+ m / z 721.4285; measured value: 721.4292.

[0166] In Example 3, a complex 7a conjugated to glutathione (GSH) was obtained with a conversion rate of 25% and a selectivity of 24%. In this example, peptides of FCPF, RGDFC, cyclic peptide RGDFK (MPa), DEVDGC, biotin-labeled Biotin-GSH, and protein-labeled DEVDGC-FITC were all able to conjugate with 6a, yielding the corresponding β-peptidyl porphyrin metal complexes.

[0167] Among them, FCPF with a π-clamp structure undergoes a coupling reaction with 6a to obtain a single coupling product 7b, which has a high conversion rate (58%) and selectivity (58%).

[0168] Compared to 7b, 7c, which contains cyclic peptides, has a higher conversion rate of 64%, but its selectivity is reduced to 44%.

[0169] Furthermore, the conversion rates and selectivity of peptides containing C-terminal cysteine ​​in 7e and 7g are similar to those in 7a. However, the peptide chain in 7f is a GSH modified with biotin, which has a larger spatial volume, and its conversion rate of 42% and selectivity of 37% are significantly improved compared to 7a.

[0170] These results indicate that by modulating the peptide sequence and steric hindrance interactions, the conversion and selectivity of meso-substituted phenyl-β-octafluoroporphyrin metal complex (6a) can be improved, that is, the coupling efficiency of meso-substituted phenyl-β-octafluoroporphyrin metal complex (6a) with thiol biomolecules (such as peptides) can be improved.

[0171] Solutions of 7a–7f dimethyl sulfoxide with a concentration of 10 μmol / L were prepared, and their normalized fluorescence emission spectra were obtained. The fluorescence emission spectra are shown below. Figure 1 As shown. (Excitation wavelength 405nm, testing instrument: Edinburgh FLS1000 steady-state transient fluorescence-electroluminescence module)

[0172] Example 5

[0173] Stock solutions of 6a and 7d with a concentration of 1 mmol / L were prepared using dimethyl sulfoxide (DMSO) as the solvent. The concentrations of the stock solutions of 6a and 7d were determined by the concentration of Yb. 3+ Concentrations were measured. The stock solutions were diluted to a final concentration of 20 μmol / L using complete growth medium (DMEM, Invitrogen, catalog number C11995500BT). HepG2 liver cancer cells were added to the above-mentioned complete medium containing 7d and 6a cells, respectively, and incubated for 0.5 hours. The HepG2 liver cancer cells were then washed twice with PBS buffer for near-infrared fluorescence imaging.

[0174] Hepatocyte LO2 cells were added to complete culture medium containing 6a or 7d cells (6a or 7d concentration of 20 μmol / L) and incubated for 0.5 hours, followed by washing twice with PBS buffer. Fluorescence imaging was performed on several cultures of hepatocyte LO2 cells in complete culture medium containing 7d cells.

[0175] Near-infrared (NIR) confocal images were acquired using the ISS Alba5 FLIM / FFS confocal system (ISS Inc.) and ISS Vista5 software. This system was connected to a Nikon TE2000 inverted microscope equipped with a Nikon 60X / 1.2NA water immersion objective.

[0176] HepG2 liver cancer cells or LO2 hepatocytes treated for 6 years or 7 days were excited using a 405nm laser. Fluorescence imaging was performed using both a Semrock 776nm and a 980nm long-pass filter. The specific fluorescence imaging details are as follows: Figure 2 (The scale bar in the figure is 10 μm) As shown, Group A consists of HepG2 liver cancer cells after 6 days of treatment; Group B consists of HepG2 liver cancer cells after 7 days of treatment; and Group C consists of LO2 hepatocytes after 7 days of treatment.

[0177] from Figure 2 It can be seen that 7d is mainly distributed on the cell membrane of HepG2. Figure 2 Group B), 6a is mainly distributed inside HepG2 cells ( Figure 2Group A). ​​Fluorescence assays of hepatocytes containing 7d showed that the 7d content on the cell membrane of hepatocyte LO2 cells was very low, and its fluorescence intensity was much lower than that of HepG2 cells.

[0178] ICP-MS (Inductively Coupled Plasma Mass Spectrometry) was used to detect Yb in LO2 of HepG2 hepatocellular carcinoma cells and healthy hepatocytes, respectively. 3+ The content was used to detect the cellular uptake at 6a and 7d, and the measurement results were as follows: Figure 3 As shown.

[0179] from Figure 3 It can be seen that Yb in HepG2 liver cancer cells treated for 7 days... 3+ The content was much higher than LO2, which is consistent with the above fluorescence imaging results. Meanwhile, Yb in 6a-treated HepG2 liver cancer cells... 3+ Cellular uptake is much less than that in LO2.

[0180] In summary, compared to 6a, 7d is more selective in targeting HepG2 liver cancer cells. It is well known that HepG2 liver cancer cells possess α... v The characteristic of β3 integrin overexpression is that the cyclic peptide RGDFK(MPa) in 7 days has the ability to target α v The properties of β3 integrin enable it to target and recognize α v HepG2 liver cancer cells overexpressing β3 integrin were targeted to achieve 7 days of targeted therapy.

[0181] Example 6

[0182] ICP-MS was used to detect Yb in HepG2 liver cancer cells. 3+ The content was used to detect the amount of cellular uptake at 6a and 7d, and the measurement results are as follows: Figure 4 As shown.

[0183] from Figure 4 It is known that using 100 μmol / L sodium azide (NaN3) as an inhibitor of respiration in HepG2 liver cancer cells will simultaneously reduce their uptake by cells at 7d and 6a.

[0184] Notably, under the same conditions, after incubation with 20 μmol / L iRGD peptide for half an hour, the uptake of 7d by HePG2 hepatocellular carcinoma cells highly expressing αvβ3 integrin was inhibited by iRGD peptide, while the uptake of 6a by HePG2 cells under the same conditions was not inhibited by iRGD peptide. This is because iRGD peptide binds to αvβ3 integrin more readily than the peptide RGDFK (MPa). Therefore, it can be concluded that the peptide RGDFK (MPa) enhances the uptake of 7d by cells highly expressing αvβ3 integrin. v Targeting of β3-mediated HePG2 hepatocellular carcinoma cells.

[0185] Example 7

[0186] 7-day stock solutions with a concentration of 1 mmol / L were prepared using dimethyl sulfoxide (DMSO) as the solvent. The concentration of the 7-day stock solutions was determined by the concentration of Yb. 3+ Concentrations were measured. The stock solutions were diluted to a final concentration of 20 μmol / L with complete growth medium. HeLa cells and HepG2 hepatocellular carcinoma cells were added to the aforementioned complete medium containing 7 days of culture and incubated for 0.5 hours. Then, the HeLa cells and HepG2 hepatocellular carcinoma cells were washed twice with PBS buffer for fluorescence imaging, using the same fluorescence imaging method as in Example 5. The test results are as follows: Figure 5 As shown (the scale bar in the figure is 125μm).

[0187] exist Figure 5 As can be seen, compared with the normal expression of α v Compared to β3 integrin-containing HeLa cell spheroids, in α v High-intensity near-infrared fluorescence of 7d was observed in HepG2 liver cancer cells overexpressing β3 integrin, indicating that 7d can target α v Cells overexpressing β3 integrin.

[0188] Example 8

[0189] Mice were intravenously injected with 100 μL of a 3.3 mg / mL 7-day aqueous solution. Eight hours later, fluorescence imaging was performed on the mice, and the results are as follows: Figure 6 As shown. Both 900nm and 1000nm long-pass filters were used. The excitation wavelength was 532nm. The exposure time was 50ms.

[0190] from Figure 6 As can be seen, 8 hours after mice were injected with the aqueous solution for 7 days, the tumor boundary could be seen through Yb 3+ The unique NIR II fluorescence distinguishes it, indicating that it has good tumor-targeting fluorescence imaging capability in vivo for 7 days, and its fluorescence emission wavelength is 950-1100nm (e.g., Figure 1 As shown, it can perform imaging in the NIR II region (1000-1700nm), and its fluorescence imaging has a deeper tissue penetration depth and higher spatial resolution, which greatly improves its accuracy as a photosensitizer for imaging.

[0191] Subsequently, the mice were euthanized, and in vitro NIR II fluorescence imaging was performed on the heart, liver, spleen, lungs, kidneys, and tumors. The tumors showed very strong fluorescence, second only to the liver. The high fluorescence intensity in the liver is because it is the primary metabolic organ at day 7. Specific fluorescence imaging details are as follows: Figure 6 As shown.

[0192] It is worth noting that the 7d provided by this invention is the first Yb biological probe for tumor imaging, which has high luminescence activity for fluorescence bioimaging of the NIR II region in vivo and in vitro tissues.

[0193] The present invention has been described in detail above with reference to specific embodiments and / or exemplary examples, as well as the accompanying drawings. However, these descriptions should not be construed as limiting the present invention. Those skilled in the art will understand that various equivalent substitutions, modifications, or improvements can be made to the technical solutions and embodiments of the present invention without departing from the spirit and scope of the invention, and all such modifications and improvements fall within the scope of the present invention. The scope of protection of the present invention is defined by the appended claims.

Claims

1. A method for preparing a product containing... β Methods for developing octafluoroporphyrin metal complexes with peptide groups. The containing β In octafluoroporphyrin metal complexes with a peptide group, the peptide group interacts with the thiol group. β - Coupling with octafluoroporphyrin-based metal complexes; The polypeptide used for coupling contains thiol groups, and the peptide group is in... β Position and β - Coupling with octafluoroporphyrin-based metal complexes; The peptide group is a polypeptide group that serves as a substituent. The containing β The octafluoroporphyrin metal complex with a peptide group has the structure shown in formula (1): in, M is a metallic element or a metallic element with ligands, wherein the metallic element is one of zinc, ytterbium, gadolinium, lutetium, erbium, and neodymium; The ligand is an acetylacetone ligand, a pyrazolborane ligand, or a phosphonooxy ligand; R are each independently selected from group 1 to group 11, and are identical to each other: , , , , , , , , , or , where Ph is phenyl; G represents a polypeptide group containing a mercaptopropionic acid group, a polypeptide group containing a cysteine ​​residue, or a polypeptide group containing a mercaptoacetic acid group. The method utilizes meso -Substituted phenyl- β -Octafluoroporphyrin metal complexes and peptides were prepared via a thiol nucleophilic substitution coupling reaction. The method specifically includes the following steps: Step 1, Preparation meso -Substituted phenyl- β -Octafluoroporphyrin metal complex; Step 2, meso -Substituted phenyl- β - Octafluoroporphyrin metal complexes and peptides are added to a biological buffer solution to react and obtain a reaction solution; Step 3: Post-process the reaction solution to obtain a solution containing... β Octafluoroporphyrin metal complexes with a peptide group. Step 1 specifically includes the following sub-steps: Sub-step a, meso -Substituted phenyl- β -Octafluoroporphyrin; The meso -Substituted phenyl- β In -octafluoroporphyrin, the substituted phenyl group is selected from haloalkylphenyl, phenyl or ether substituted phenyl containing sulfonic acid group and halo group; The meso -Substituted phenyl- β -Octafluoroporphyrin is meso -haloalkylphenyl- β - Octafluoroporphyrin, meso -(sulfonic acid halophenyl)- β -octafluoroporphyrin or meso -(ether-substituted phenyl)- β -Octafluoroporphyrin; Sub-step b, preparation meso -Substituted phenyl- β -Octafluoroporphyrin metal complex; The meso -Substituted phenyl- β - Octafluoroporphyrin metal complexes are composed of meso -Substituted phenyl- β -Octafloporphyrin is obtained by combining it with a metal compound.

2. The method according to claim 1, characterized in that, The ligand is the Kläui ligand.

3. The method according to claim 1, characterized in that, Includes β Octafluoroporphyrin metal complexes with peptide groups are selected from 7a to 7g: 、 、 、 、 、 、 ， Among them, Yb coordinates with the oxygen in the three "P=O" bonds in L.

4. The method according to claim 1, characterized in that, The substituted phenyl group is a chloroalkylphenyl, a bromoalkylphenyl, an iodoalkylphenyl, a group 10, or a group 11.

5. The method according to claim 1 or 4, characterized in that, The condensation of 3,4-difluoropyrrole with p-haloalkylbenzaldehyde, m-haloalkylbenzaldehyde, or oro-haloalkylbenzaldehyde yields... meso -haloalkylphenyl- β -Octafluoroporphyrin is prepared by first complexing it with a metal compound, and then reacting it with one of the following compounds: a nitrogen-containing heterocyclic compound, a tertiary amine compound, a tertiary phosphine compound, a thioether compound, and sulfuric acid. meso -Substituted phenyl- β -Octafluoroporphyrin metal complex; 3,4-Difluoropyrrole is condensed with o-dihalobenzaldehyde or m-ether benzaldehyde to obtain meso -(sulfonic acid halophenyl)- β -octafluoroporphyrin or meso -(ether-substituted phenyl)- β -Octafluoroporphyrin, which is then complexed with a metal compound to obtain meso -Substituted phenyl- β -Octafloporphyrin metal complex.

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