CN109678888A - Oxazine compound and application thereof - Google Patents

Oxazine compound and application thereof Download PDF

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Publication number
CN109678888A
CN109678888A CN201811635409.5A CN201811635409A CN109678888A CN 109678888 A CN109678888 A CN 109678888A CN 201811635409 A CN201811635409 A CN 201811635409A CN 109678888 A CN109678888 A CN 109678888A
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compound
cell
oxazine compound
mitochondria
alkyl
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CN109678888B (en
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樊江莉
姚起超
杜健军
孙文
龙飒然
邵堃
王静云
彭孝军
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DALIAN KERONG BIOLOGICAL TECHNOLOGY Co Ltd
Dalian University of Technology
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DALIAN KERONG BIOLOGICAL TECHNOLOGY Co Ltd
Dalian University of Technology
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Abstract

Oxazine compound and application thereof, the oxazine compound have the structure of general formula F.The oxazine compound with general formula F of aforementioned present invention has specificly-response to mitochondria, can rapidly enter cell, be combined rapidly with mitochondria and issue the fluorescence compared with strong signal.No matter in fixed cell or in living cells experiment, preferable specific recognition is embodied to mitochondria and is marked.Furthermore the compound of the present invention is a kind of light/sound photodynamic activity organic molecule for having and having photosensitive and sound quick characteristic while near infrared absorption transmitting.Such compound absorption maximum and launch wavelength are all larger than 690nm, triplet state high conversion rate, under illumination or ultrasound condition, can greater efficiency generate active oxygen species, there is preferable lethal effect for cancer cell and cancerous tissue, hardly normal tissue increases toxic side effect while reaching light/sound dynamic therapy to tumour.

Description

Oxazine compound and application thereof
Technical field
The present invention relates to a kind of novel oxazines compound, its synthesis and its newly answering in biospecificity recognition detection With.Belong to Anti-Cancer Drug Design, synthesis and application field simultaneously.Synthesis more particularly to novel oxazines class light/sound sensitiser and its Specific recognition to tumour and the new opplication in diagnosis and treatment.
Background technique
Cell (cell) is to be surrounded by film, and can be carried out the primary matter group independently bred, and is that human body composition is most basic Structure and function unit, an adult is probably made of 100,000,000,000,000 cells, the various physiology and pathologic process of human body all with Cell proliferation metabolism is closely related.Since 20th century, with deepening continuously for Celluar and Molecular Biology research, science Worker concentrates on more sight in organelle research level.Organelle be have in cytoplasm independent physiological function and The morphosis unit of certain chemical composition, by taking zooblast as an example, mainly have nucleus, mitochondria, lysosome, endoplasmic reticulum, The organelles such as golgiosome
Mitochondria (Mitochondrion) is that Germany scientist Altmann in 1894 has found for the first time in zooblast, It is the biggish a kind of organelle of volume that can see under light microscopic, has permeability, can expand in hypotonic solution, hypertonic It is shunk in solution.Because its form is in line or graininess, thus it is named as mitochondria.Studies have shown that many serious diseases with Mitochondria abnormal behavior is related.Current most of mitochondria positioning dyestuffs are all based on the characteristic of the negative film potential of mitochondria, and line A lot of other function and mechanism ofs of plastochondria are not known.Therefore the research to mitochondria, such as mitochondrial DNA (mtDNA) imaging, activity Oxygen response, ion (Ca2+, Cu+Deng) measurement, effect of the mitochondria in Apoptosis etc. be still scientists concern hot spot. If form and physiology course of mitochondria etc. are abnormal, great threat, therefore realization pair will be generated to the health of organism The real-time monitoring of mitochondria, prevention and treatment to various major diseases have practical significance.
The great function that fluorescence probe plays in the fields such as biology, medical treatment is increasingly becoming the research hot topic of people.From glimmering At the beginning of photoinitiator dye molecule is found between till now more than 200 years, people have been achieved for numerous research achievements.Development more at Ripe fluorescent dye includes flower cyanine type dye, fluorine boron pyrroles, cumarin, rhodamine, fluorescein, Nile blue etc., people by A series of functional groups are connected on its parent ring to be modified, and have been synthesized a variety of fluorescent probe molecules with detection function, have been used for Target molecule realizes visualizing monitor.Luminescent dye molecule has been applied to luminescent material and sensor, environment pollution detection, life The various fields such as object research, medical diagnosis.It is noted that fluorescence probe plays an important role in terms of medical treatment & health. They can enter cell, be identified positioning to intracellular various subcellular structures, intuitive to distinguish normal cell and lesion Cell, so that it is determined that illness root, is conducive to immunotherapy targeted autoantibody disease.
So far, in this direction, people have been achieved for certain achievement, some mitochondria fluorescence probes and its thin The outstanding advantages being imaged in born of the same parents gradually developed.But in the probe currently reported, generally existing dyeing concentration Greatly, incubation time is long, the deficiency that probe cytotoxicity is big etc. needs to be improved.
Photodynamic action refers in photosensitizer presence, under the action of light, organic body cell or biomolecule occurs Function or metamorphosis lead to cellular damage and necrosis effect, the participation that early stage this effect must be aerobic, so again when serious Claim photosensitizer-oxidation, this effect be referred to as optical sensibilization in chemistry, in biology and medically referred to as light power Effect, the method cured the disease with photodynamic action, referred to as photodynamic therapy (photodynamic therapy, PDT).Light power is treated Method is the new disease treatment means of one kind based on the interaction of light, photosensitizer and oxygen, photosensitizer (Visudyne Object) research be influence optical dynamic therapy prospect key point.Photosensitizer is some special chemical substances, basic role It is transmitting energy, it can absorb photon and be excited, and the luminous energy of absorption is passed to rapidly to the molecule of another component, make it It is excited and photosensitizer itself returns to ground state.As first photosensitizer Porfimer Sodium is in 1993--1997 Nian Mei State, Canada, European Union, Japan and South Korea go through to list successively, and research, the development and application in the field PDT are active rapidly. With new optical dynamic therapy medicine research and develop successfully and the raising of laser equipment technology, PDT have welcome unprecedented hair Open up peak.In the world, approved lists or just in nearly ten kinds of the new photosensitizer of clinical research.Meanwhile PDT is also used for non-swell Tumor type disease, as condyloma acuminatum, psoriasis, nevus flammeus, rheumatoid arthritis, fundus flavimaculatus lesion, angioplastic are postoperative again The treatment of the diseases such as narrow.Domestic representational photosensitizer mainly has: Shanghai Fudan Zhangjiang biomedical Co., Ltd The Ai La of development & production (5-ALA, external application hydrochloric acid ammonia ketone valeric acid dissipate).
Currently, it is applied to clinical light/sound sensitiser mainly using porphyrin and phthalocyanine-like compound as main representative, although such Compound achieves in terms of oncotherapy compared with ten-strike, but still remains many defects, such as: therapy system composition ratio Unstable, slow in human body intracellular metabolite, maximum excitation wavelength is shorter, and light toxic side effect etc. easily occurs.These deficiencies seriously affect The actual effect and clinical application of photodynamic therapy.Preparation and application study for photosensitizer have had certain in the world Guiding theory can play directive function, but preparation and application study for sound sensitiser, do not have relevant theoretical direction, And it is even more very few to can be applied to clinical sound sensitiser.Therefore suitable light/sound sensitiser is designed, had for tumour diagnosis and treatment Very big progradation.
Summary of the invention
The purpose of the present invention is to provide a kind of with the quick characteristic of light/sound can specific recognition mitochondria fluorescence probe Molecule.For this purpose, present invention firstly provides a kind of oxazine compound, the structure with general formula F:
In general formula F,
The A is selected from oxygen (O), sulphur (S), selenium (Se) and tellurium (Te)；
The R1、R2And R3It is each independently selected from the group of hydrogen, formula i- X, C1-12Alkyl and C1-12Substitution alkane Base；
The substitution alkyl is arbitrarily replaced by following radicals: halogen ,-OH ,-COOH ,-NO2、-SO3H (sulfonic group) ,- OCH3、-OC2H5、-OC3H7、-OC4H9、-COOCH3、-COOC2H5、-COOC3H7、-COOC4H9、-COOC5H11、-COOC6H13、- NR4R5Or-CONR6R7；R therein4、R5、R6、R7It is each independently selected from hydrogen and C1-6Alkyl；
The X is selected from dihydrogen phosphate, bisulfate ion, nitrate anion, chlorine anion, bromine anion, iodine anion or height Chlorate anions.
The oxazine compound with general formula F of aforementioned present invention to mitochondria have specificly-response, can quickly into Enter cell, combined rapidly with mitochondria and issues the fluorescence compared with strong signal.No matter in fixed cell or in living cells experiment, Preferable specific recognition is embodied to mitochondria to mark.Further by a series of performance tests, it is found that the probe molecule exists With the maximum absorption wavelength (about 690nm) and maximum emission wavelength (about 710nm) of near-infrared, the excitation of long wavelength in aqueous systems And launch wavelength light energy is lower, and it is smaller to histiocytic damage, and photopermeability is good, histiocytic autofluorescence pair Its interference is smaller.And its corresponding fluorescence intensity of fluorescence quantum yield in a variety of different organic solvents is corresponding.And And such compound has the water solubility of certain level, while having good permeability of cell membrane, and bio-toxicity, light poison Property, photobleaching are lower.Its spectral region and the spectral region of biological sample have sufficiently large difference.Furthermore such chemical combination Object has preferable photostability, can also be stabilized under physiological pH condition, is conducive to it applied to hair in organism Wave fluorescent probe function.
Therefore, the oxazine compound that is designed to provide of another aspect of the present invention is preparing near-infrared fluorescent spy Application in needle, the near infrared fluorescent probe are Mitochondrially targeted fluorescence probes.Prepare resulting near infrared fluorescent probe It can be used for the fluorescence imaging of cell or tissue Mitochondria, and the form of mitochondria can be monitored in real time, and can be applied to surpass Resolution mode.
On the other hand, heretofore described and oxazine compound be a kind of tool while there is near infrared absorption transmitting Light/sound photodynamic activity organic molecule of the standby quick characteristic of photosensitive and sound.Such compound absorption maximum and launch wavelength are all larger than 690nm, triplet state high conversion rate, under illumination or ultrasound condition, can greater efficiency generate active oxygen species, for cancer cell and Cancerous tissue has preferable lethal effect, and hardly normal tissue increases while reaching light/sound dynamic therapy to tumour Toxic side effect.Therefore, another aspect of the present invention provides the oxazine compound and is preparing the application in light/sound sensitiser.It should Light/the sound sensitiser is near-infrared long-wavelength fluorescent probe, the label for tumour cell.Oxazines class of the present invention Object is closed in addition to small molecule dosing techniques can be carried out, can also be self-assembled into nanoparticle or is coated with by other materials into nanoparticle It is administered operation, nanoscale effective range is 1-1000nm.
Detailed description of the invention
13 width of attached drawing of the present invention:
Fig. 1 is the general structure F of oxazine compound of the invention.
Fig. 2 is result figure of the probe compound F-1 in breast cancer cell MCF-7 labelling experiment, in which: Fig. 2 (a) is F-1 Channel；Fig. 2 (b) is cell light field figure；Fig. 2 (c) is (a) and the hybrid channel (b).
Fig. 3 is mitochondria common location experimental result picture of the probe compound F-1 in breast cancer cell MCF-7, in which: figure 3 (a) be the commercialization channel dyestuff MitoTracker GreenFM；Fig. 3 (b) is the channel F-1；Fig. 3 (c) is that (a) is mixed with (b) Channel；Fig. 3 (d) is Intracellular signals distribution map and common location coefficient.
Fig. 4 is lysosome common location experimental result picture of the probe compound F-1 in breast cancer cell MCF-7, in which: figure 4 (a) be the commercialization channel dyestuff LysoTracker GreenFM；Fig. 4 (b) is the channel F-1；Fig. 4 (c) is that (a) is mixed with (b) Channel；Fig. 4 (d) is Intracellular signals distribution map and common location coefficient.
Fig. 5 is labelling experiment result figure of the probe in living body Kunming mouse.
Fig. 6 is result figure of the probe compound F-2 in breast cancer cell MCF-7 labelling experiment.Wherein, Fig. 6 (a) F-2 is logical Road；Fig. 6 (b) is cell light field figure；Fig. 6 (c) is (a) and the hybrid channel (b).
Fig. 7 is the mitochondria common location experimental result picture in the breast cancer cell MCF-7 of probe compound F-2.Wherein, scheme 7 (a) be the commercialization channel dyestuff MitoTracker GreenFM；Fig. 7 (b) is the channel F-2；Fig. 7 (c) is that (a) is mixed with (b) Channel；Fig. 7 (d) is Intracellular signals distribution map and common location coefficient.
Fig. 8 is the result figure of lysosome common location experiment of the probe compound F-2 in breast cancer cell MCF-7.Wherein: Fig. 8 (a) is the commercialization channel lysosome dyestuff LysoTracker GreenFM；Fig. 8 (b) is the channel F-2；Fig. 8 (c) be (a) with (b) hybrid channel；Fig. 8 (d) is Intracellular signals distribution map and common location coefficient.
Fig. 9 is that Mitochondrial Shape of the probe compound F-2 in breast cancer cell MCF-7 shows experimental result picture.Wherein, Fig. 9 (a) is the mitochondria of normal filigree shape；Fig. 9 (b) be mitochondria by slight damage when, most of Mitochondrial Shapes remain to Keep filament shape；When Fig. 9 (c) is that damage aggravates that mitochondria is made to start expansion, form gradually becomes round point shape；Fig. 9 (d) be by When damaging very serious, Mitochondrial Shape is empty balloon-shaped；Fig. 9 (A) is the partial enlarged view of (a)；Fig. 9 (B) is the partial enlargement of (b) Figure；Fig. 9 (C) is the partial enlarged view of (c)；Fig. 9 (D) is the partial enlarged view of (d).
Figure 10 is labelling experiment result figure of the probe compound F-2 in living body Kunming mouse.
Figure 11 is that Mitochondrial Shape of the probe compound in breast cancer cell MCF-7 shows experimental result picture.
Figure 12 is singlet oxygen yield measurement experiment result figure under probe compound F-3 illumination condition.
Figure 13 is cell in vitro anticancer test result figure under probe compound F-3 illumination condition.
Specific embodiment
Unless otherwise stated, term used herein has following meanings.
Term " alkyl " used herein includes straight chained alkyl and branched alkyl.In the feelings for being not described as " replacing alkyl " Under condition, term " alkyl " is often referred to unsubstituted alkyl.Such as refer to that single alkyl such as " propyl ", then only refers in particular to straight chained alkyl, such as Refer to that single branched alkyl such as " isopropyl ", then only refers in particular to branched alkyl.For example, " C1-6Alkyl " includes C1-4Alkyl, C1-3Alkane Base, methyl, ethyl, n-propyl, isopropyl and tert-butyl.Similar rule is also applied for other bases used in this specification Group.
Term " halogen " used herein includes fluorine, chlorine, bromine and iodine.
The present invention is to have the structure of general formula F, such as Fig. 1 about a kind of oxazine compound first:
In the general formula F, the A is selected from oxygen (O), sulphur (S), selenium (Se) and tellurium (Te).
The R1、R2And R3Selectable group range is extensive, and the selection in of the present invention and range is to molecule Charge characteristics and hydrophilic and oleophilic coefficient do not generate substantive influence.The R1、R2And R3Respectively can independently selected from hydrogen, The group of formula i-x, the alkyl or substitution alkyl of carbon atom number 1-12；It is wherein described and alkyl or replace the carbon atom number of alkyl It is preferred that 1-8, more preferable 1-6.
The upper substitution alkyl is arbitrarily replaced by following radicals: halogen ,-OH ,-COOH ,-NO2、-SO3H (sulfonic acid Base) ,-OCH3、-OC2H5、-OC3H7、-OC4H9、-COOCH3、-COOC2H5、-COOC3H7、-COOC4H9、-COOC5H11、- COOC6H13、-NR4R5Or-CONR6R7；R therein4、R5、R6、R7It is each independently selected from hydrogen and C1-6Alkyl.Preferably, described R4、R5、R6、R7It is hydrogen.
In specific embodiment, R1And R2Middle one is H；Further preferably, the R3It also is H.
In some specific embodiments, R1And R2Middle one is H, another, is the group selected from formula i-x.This kind ofization Close the function that object has tumour identification due to the group of formula i-x.Therefore it can be used for the positioning of tumour cell and quick as light/sound Agent application.
In the general formula F, X therein be selected from dihydrogen phosphate, bisulfate ion, nitrate anion, chlorine anion, bromine bear from Son, iodine anion or perchlorate.
On the basis of the mitochondria specific recognition capability of each compound is screened and is compared, it is provided by the invention most In preferred embodiment, oxazine compound of the present invention be selected from F-1, F-2, F-3, F-4, F-5, F-6, F-7, F-8, F-9, F-10, F-11 or F-12:
Oxazine compound of the invention has specificly-response to mitochondria, cell can be rapidly entered, with nucleus Interior mitochondria combines rapidly and issues the fluorescence compared with strong signal.No matter in fixed cell or living cells experiment, to line grain Body embodies preferable specific recognition label.Further by a series of performance tests, find the probe molecule in aqueous systems Maximum absorption wavelength (about 691nm) and maximum emission wavelength (about 711nm), the excitation of long wavelength and transmitted wave with near-infrared Long light energy is lower, smaller to histiocytic damage, and photopermeability is good, histiocytic autofluorescence it is interfered compared with It is small.And its corresponding fluorescence intensity of fluorescence quantum yield in a variety of different organic solvents is corresponding.And such is changed The water solubility that object has certain level is closed, while there is good permeability of cell membrane, and bio-toxicity, phototoxicity, light float Bai Xingjun is lower.Its spectral region and the spectral region of biological sample have sufficiently large difference.Furthermore such compound have compared with Photostability well can be also stabilized under physiological pH condition, be conducive to it and visited applied to performance fluorescence in organism Needle function.Excellent photostability makes such compound have preferable super-resolution microscopy work ability.
Based on this, the present invention is by oxazines class of the present invention to the specific technical solution of the application of the compound Compound is used to prepare the near infrared fluorescent probe product of Mitochondrially targeted fluorescence probe class, is used for cell or tissue Mitochondria Fluorescence imaging.Heretofore described near infrared fluorescent probe is used in aqueous solution, buffer solution carry out mitochondria special Opposite sex label and detection.
Compound with general formula F described above, preparation method is first public, and those skilled in the art should Compound of the present invention can be completed in conjunction with the technical information of related fields and the basic theories of organic synthesis and technology It obtains.The preparation method of following oxazine compounds described in this specification provides a kind of specific side of such compound synthesis Case, but it is not construed as the restriction to it.
It is heretofore described and oxo oxazine compound pass through following methods and synthesize: formed using anthracene or derivatives thereof Azo-compound be condensed in the DMF containing acid with 8- hydroxyl julolidine, target oxazine dye is prepared.The synthetic method work Skill is succinct, high conversion rate.More specifically, general formula compound F synthetic route of the present invention can indicate are as follows:
The preparation method of the compound of general formula F represented by above-mentioned route includes the following steps:
(1) in hydrochloric acid acid system, chlorination is to the compound of nitro diazobenzene and Formulas I according to molar ratio 1:1 25~35 It is reacted 0.5~2 hour under the conditions of DEG C, preparation formula II compound；
(2) Formula II compound and the long lourie pyridine of 8- hydroxyl according to molar ratio 1:1 in acid DMF in 135~145 DEG C of conditions It is lower to react the compound for preparing general formula F for 2~4 hours.
It is heretofore described and sulfur selenium tellurium generation oxazine compound pass through following methods synthesize: using arylamine or its spread out The azo-compound that biology is formed is condensed in the DMF containing acid with 8- hydroxyl julolidine, and target oxazine dye is prepared.The conjunction It is succinct at method and process, high conversion rate.More specifically, general formula compound F synthetic route of the present invention are as follows:
The preparation method of the compound of general formula F represented by above-mentioned route includes the following steps:
(1) in hydrochloric acid acid system, chlorination is to the compound of nitro diazobenzene and Formulas I according to molar ratio 1:1 25~35 It is reacted 0.5~2 hour under the conditions of DEG C, preparation formula II compound；
(2) Formula II compound and the long lourie pyridine of 8- hydroxyl according to molar ratio 1:1 in acid DMF in 135~145 DEG C of conditions It is lower to react the compound for preparing general formula F for 2~4 hours.
It is of the present invention to have following advantages by the near infrared fluorescent probe of parent of oxazines:
The compound has the water solubility of certain level, while having good permeability of cell membrane.
The compound has specificity, specific recognition for mitochondria；
The compound have excellent fluorescence property, applied to biological sample imaging when with low biological photobleaching, Light injury and bio-toxicity, and the fluorescence signal generated can penetrate deeper biological tissue；
The fluorescence emission wavelengths of the compound moieties are greater than 700nm, can be used for living animal imaging；
The compound is used for the label of tumour and tumour cell and tissue, and good mitochondrial markers may be implemented, and It is avoided that interference of the external environmental factor to fluorescence intensity；
The compound can be applied to super-resolution microscope imaging；
The compound side effect is small, and raw material is easy to get, and structure is simple, easily prepared, easy industrialization；
In consideration of it, near infrared fluorescent probe compound of the present invention can be used for tumour and non-tumor cell and tissue is marked Note.Other than being directly used in the dyeing of tumour and non-tumor cell and tissue in form described herein, containing of the invention The composition of near infrared fluorescent probe compound can be used for the dyeing of tumour cell and tissue.It should be wrapped in the composition Containing one of a effective amount of two-photon fluorescence probe compound provided by the present invention.Furthermore it is also possible to include biological sample dyeing Required other components, such as solvent, pH adjusting agent etc..These components are all that current row is known in the art.Above-mentioned composition can To exist as an aqueous solution, or can be to exist before use with other suitable forms that water is formulated as solution.
The present invention also provides the near infrared fluorescent probe compound label tumour cells for using aforementioned present invention and tissue life The method of object sample, this method include the steps that contacting the compound with biological sample.Term used herein " connects Touching " may include contacting in solution or solid phase.
Following non-limiting embodiments can with a person of ordinary skill in the art will more fully understand the present invention, but not with Any mode limits the present invention.
Embodiment 1: probe compound F-1 is prepared
(1) synthesis of intermediate 1-II
In hydrochloric acid acid system, chlorination is to the compound of nitro diazobenzene and 1-I according to molar ratio 1:1 in 25~35 DEG C of items It is reacted 0.5~2 hour under part, end of reaction, brick-red solid powder crude product is obtained after filtering and washing operates and obtains formula 1- The compound of II, yield 95%.
(2) synthesis of compound F-1
The intermediate 1-II and Se that above-mentioned reaction (1) is prepared is added to the round bottom containing DMF for julolidine to burn In bottle, 1mL perchloric acid solution is instilled.It is added dropwise, system stops reaction after stirring 2.5h, separates (dichloro through silica gel column chromatography Methane: methanol=8:1) purify tool metallic luster blue-green acicular crystal target-probe compound F-1, yield 78.2%.
1H NMR(400MHz,DMSO-d6) δ 9.69 (s, 1H), 9.18 (s, 1H), 9.09 (s, 1H), 8.29 (d, J= 7.9Hz, 1H), 8.11 (d, J=8.1Hz, 1H), 7.84-7.71 (m, 2H), 7.39 (s, 1H), 6.73 (s, 1H), 3.57-3.44 (m,3H),2.85(s,2H),2.80–2.72(m,2H),1.95(s,3H).HRMS(ESI,HRDFMagSec)m/z 456.0980 (caled for C26H22N3Se+:456.0978)
Embodiment 2: labelling experiment of the probe compound F-1 in breast cancer cell (MCF-7)
Using the compound F-1 synthesized in embodiment 1,
F-1-DMSO solution is added in the MCF-7 cell containing 2mL culture medium and is shaken, laser confocal microscope is used Imaging.Representative area is chosen, is observed with oil mirror (60 ×), in triplicate.As a result as shown in Figure 2, in which: Fig. 2 (a) F-1 is logical Road；Fig. 2 (b) is cell light field figure；Fig. 2 (c) is (a) and the hybrid channel (b).It can be seen that F-1 molecule can be to cell color.
Embodiment 3: mitochondria common location experiment of the probe compound F-1 in breast cancer cell (MCF-7)
The compound F-1 synthesized using embodiment 1,
F-1-DMSO solution is added in the MCF-7 cell containing culture medium (cell in advance at 37 DEG C, 5%CO2It is lower to incite somebody to action Commercialization dyestuff mitochondrial dye MitoTracker Green is addedFMIt is incubated for 20 minutes in culture medium.Then, PBS concussion drift 5min × 3 are washed, cell culture medium is added) concussion, it is imaged with laser confocal microscope.Representative area is chosen, oil mirror is used (60 ×) it observes, in triplicate.As a result as shown in figure 3, Fig. 3 (a) is commercialization dyestuff MitoTracker GreenFMChannel；Figure 3 (b) be the channel F-1；Fig. 3 (c) is (a) and the hybrid channel (b)；Fig. 3 (d) is Intracellular signals distribution map and common location coefficient. It can be seen that F-1 molecule is distributed mainly in cell mitochondrial.
Embodiment 4: lysosome common location experiment of the probe compound F-1 in breast cancer cell (MCF-7)
F-1-DMSO solution is added to the MCF-7 cell containing culture medium by the compound F-1 synthesized using embodiment 1 In (cell in advance at 37 DEG C, 5%CO2Lower will be added is commercialized dyestuff lysosome dyestuff LysoTracker GreenFMIn culture It is incubated for 20 minutes in base.Then, PBS concussion rinsing 5min × 3, add cell culture medium) concussion, it is micro- with laser co-focusing Mirror imaging.Representative area is chosen, is observed with oil mirror (60 ×), in triplicate.As a result as shown in figure 4, Fig. 4 (a) is commercialization Dyestuff LysoTracker GreenFMChannel；Fig. 4 (b) is the channel F-1；Fig. 4 (c) is (a) and the hybrid channel (b)；Fig. 4 (d) is Intracellular signals distribution map and common location coefficient.It can be seen that F-1 molecule is not distributed in cytase body.
Embodiment 5: labelling experiment of the probe compound F-1 in living body Kunming mouse
Living body Kunming mouse injects 10% chloraldurate (10mg/Kg) anesthesia, then sucks appropriate Isoflurane and deepen anesthesia and light Degree inhibits breathing (being reduced to movement and respiration artefacts minimum), and F-1-DMSO solution is injected into after diluting 1000 times with pure PBS Living body Kunming mouse abdomen.Living body Kunming mouse is placed in small animal imaging instrument, takes supine position in being imaged in fixed plate. As a result as shown in Figure 5, it is seen that living body biological sample painted can be imaged in F-1 molecule.
Embodiment 6: probe compound F-2 is prepared
(1) synthesis of intermediate 2-II
In hydrochloric acid acid system, chlorination is to the compound of nitro diazobenzene and 2-I according to molar ratio 1:1 in 25~35 DEG C of items It is reacted 0.5~2 hour under part, end of reaction, aubergine solid powder crude product is obtained after filtering and washing operates and obtains formula 2- The compound of II, yield 95%.
(2) synthesis of compound F-2
Intermediate 2-II and 8- hydroxyl julolidine that above-mentioned reaction (1) is prepared are added to the round bottom containing DMF In flask, 1mL perchloric acid solution is instilled.It is added dropwise, system stops reaction after stirring 2.5h, through silica gel column chromatography separation (two Chloromethanes: methanol=8:1) purify tool metallic luster green acicular crystal target-probe compound F-2, yield 65.4%.
1H NMR(400MHz,DMSO-d6) δ 9.81 (s, 1H), 9.14 (s, 1H), 9.07 (s, 1H), 8.25 (d, J= 7.8Hz, 1H), 8.10 (d, J=7.5Hz, 1H), 7.80-7.72 (m, 2H), 7.39 (s, 1H), 6.88 (s, 1H), 3.74 (s, 1H), 3.55-3.46 (m, 3H), 2.87 (s, 2H), 2.77 (s, 2H), 1.98 (m, 5H), 1.43 (t, J=7.1Hz, 3H) .HRMS (ESI,HRDFMagSec)m/z 420.2063(caled for C26H22N3O+:420.2070)
Embodiment 7: labelling experiment of the probe compound F-2 in breast cancer cell (MCF-7)
Using the compound F-2 synthesized in embodiment 6, F-2-DMSO solution is added to the MCF-7 containing 2mL culture medium It shakes in cell, is imaged with laser confocal microscope.Representative area is chosen, is observed with oil mirror (60 ×), in triplicate.Knot Fruit is as shown in Figure 6, in which: the channel Fig. 6 (a) F-2；Fig. 6 (b) is cell light field figure；Fig. 6 (c) is (a) and the hybrid channel (b).It can See that F-2 molecule can be to cell color.
Embodiment 8: mitochondria common location experiment of the probe compound F-2 in breast cancer cell (MCF-7)
F-2-DMSO solution is added to the MCF-7 cell containing culture medium by the compound F-2 synthesized using embodiment 6 In (cell in advance at 37 DEG C, 5%CO2Lower will be added is commercialized dyestuff mitochondrial dye MitoTracker GreenFMIn culture It is incubated for 20 minutes in base.Then, PBS concussion rinsing 5min × 3, add cell culture medium) concussion, it is micro- with laser co-focusing Mirror imaging.Representative area is chosen, is observed with oil mirror (60 ×), in triplicate.As a result as shown in fig. 7, Fig. 7 (a) is commercialization Dyestuff MitoTracker GreenFMChannel；Fig. 7 (b) is the channel F-2；Fig. 7 (c) is (a) and the hybrid channel (b)；Fig. 7 (d) is Intracellular signals distribution map and common location coefficient.It can be seen that F-2 molecule is distributed mainly in cell mitochondrial.
Embodiment 9: lysosome common location experiment of the probe compound F-2 in breast cancer cell (MCF-7)
F-2-DMSO solution is added to the MCF-7 cell containing culture medium by the compound F-2 synthesized using embodiment 6 In (cell in advance at 37 DEG C, 5%CO2Lower will be added is commercialized dyestuff mitochondrial dye LysoTracker GreenFMIn culture It is incubated for 20 minutes in base.Then, PBS concussion rinsing 5min × 3, add cell culture medium) concussion, it is micro- with laser co-focusing Mirror imaging.Representative area is chosen, is observed with oil mirror (60 ×), in triplicate.As a result as shown in figure 8, Fig. 8 (a) is commercialization Lysosome dyestuff LysoTracker GreenFMChannel；Fig. 8 (b) is the channel F-2；Fig. 8 (c) is (a) and the hybrid channel (b)；Fig. 8 It (d) is Intracellular signals distribution map and common location coefficient.It can be seen that F-2 molecule is not distributed in cytase body.
Embodiment 10: probe compound F-2 shows in breast cancer cell (MCF-7) Mitochondria form and tests
F-2-DMSO solution is added to the MCF-7 cell containing culture medium by the compound F-2 synthesized using embodiment 6 It is middle to use the laser irradiation different length time, damage intracellular mitochondrial in various degree.Representative area is chosen, oil mirror is used (60 ×) it observes, in triplicate.As a result as shown in figure 9, Fig. 9 (a) is the mitochondria of normal filigree shape；Fig. 9 (b) be mitochondria by When to slight damage, most of Mitochondrial Shapes are still able to maintain filament shape；Fig. 9 (c) is that damage aggravates that mitochondria is made to start to expand When, form gradually becomes round point shape；When Fig. 9 (d) is impaired very serious, Mitochondrial Shape is empty balloon-shaped；Fig. 9 (A) is (a) Partial enlarged view；Fig. 9 (B) is the partial enlarged view of (b)；Fig. 9 (C) is the partial enlarged view of (c)；Fig. 9 (D) is the office of (d) Portion's enlarged drawing.It can be seen that intracellular mitochondrial form can be monitored in real time in F-2 molecule.
Embodiment 11: labelling experiment of the probe compound F-2 in living body Kunming mouse
Living body Kunming mouse injects 10% chloraldurate (10mg/Kg) anesthesia, then sucks appropriate Isoflurane and deepen anesthesia and light Degree inhibits breathing (being reduced to movement and respiration artefacts minimum), and F-2-DMSO solution is injected into after diluting 1000 times with pure PBS Living body Kunming mouse abdomen.Living body Kunming mouse is placed in small animal imaging instrument, takes supine position in being imaged in fixed plate. The results are shown in Figure 10, it is seen that living body biological sample painted can be imaged in F-2 molecule.
Embodiment 12: probe compound F-2 super-resolution observes intracellular mitochondrial experiment
F-2-DMSO solution is added to the MCF-7 cell containing culture medium by the compound F-2 synthesized using embodiment 6 It is middle to use the micro- sem observation of super-resolution, it is clear that elongated Mitochondrial Shape.As a result as indicated at 11, it is seen that F-2 molecule tool There is the function of super-resolution intracellular mitochondrial form.It can be seen that intracellular mitochondrial form can be monitored in real time in F-2 molecule.
Embodiment 13: probe compound F-3 is prepared
(1) synthesis of intermediate 3-II
In hydrochloric acid acid system, chlorination is to the compound of nitro diazobenzene and 2-I according to molar ratio 1:1 in 25~35 DEG C of items It is reacted 0.5~2 hour under part, end of reaction, aubergine solid powder crude product is obtained after filtering and washing operates and obtains formula 3- The compound of II, yield 94%.
(2) synthesis of compound F-3
The intermediate 3-II that above-mentioned reaction (1) is prepared is added to the round bottom containing DMF with thio julolidine to burn In bottle, 1mL perchloric acid solution is instilled.It is added dropwise, system stops reaction after stirring 2.5h, separates (dichloro through silica gel column chromatography Methane: methanol=8:1) purify tool metallic luster green sheet crystal target probe compound F-3, yield 55.4%.
1H NMR(400MHz,DMSO-d6)δ8.83(s,1H),8.05–7.98(m,3H),7.62-7.58(m,2H),6.75 (s,1H),6.22(s,1H),3.64(s,1H),3.55–3.46(m,8H),2.87(s,2H),2.77(s,2H),1.96(m, 5H),.HRMS(ESI,HRDFMagSec)m/z 436.1841(caled for C28H26N3S+:436.1842)
Embodiment 14: singlet oxygen yield measurement experiment under compound F-3 illumination condition
Using the compound F-3 synthesized in embodiment 13, F-3-DMSO solution is added in methanol and is added after mixing Enter 1,3- diphenyl isobenzofuran (DPBF), adjusting DPBF concentration uses 700nm wavelength after making its absorbance value be about 1.0 Xenon source (filtered and adjusted by grating) irradiates, and constant duration measures system ultravioletvisible absorption curve.Existed according to DPBF Absorbance change at 411nm wavelength draws the correlation curve of absorbance and time, using methylene blue as reference, calculates The singlet oxygen quantum yield of compound F-3, as a result such as Figure 12.The attached drawing is shown to be inhaled with light application time mixed system UV, visible light Light spectrum change situation, can obtain according to correlation formula, and the singlet oxygen quantum yield of compound F-3 is about 0.03.
Embodiment 15: cell in vitro anticancer is tested under compound F-3 illumination condition
MCF-7 (human breast cancer cell) is planted in 96 well culture plates with 5000, every hole cell density in cell culture Cultivated in case 24 hours (37 DEG C, 5%CO2), using the compound F-3 synthesized in embodiment 13, F-3-DMSO solution is added To containing various concentration solution is configured in 10% fetal calf serum DMEM, 96 orifice plates are added in the solution prepared and are placed in cell incubator After middle incubation 30 minutes, with the red light irradiation certain time of 700nm wavelength.Irradiation finishes, and 96 hollow plates are placed in cell incubator In continue be incubated for 12 hours.Then every hole is added the 100 μ l of culture medium of the MTT containing 5mg/ml, and as being incubated in cell incubator 4 hours.The culture solution in plate hole is removed, every hole is added after 100 μ l DMSO sufficiently dissolve MTT oxidation product and is measured with microplate reader Every hole absorbance at 570nm and 630nm calculates cell survival rate, as shown in figure 13.
As can be seen from Figure 13, compound F-3 is very small to the lethal effect of cell under conditions of no illumination, almost There is no toxicity；Under conditions of having illumination, compound F-3 can generate obvious fragmentation effect with cell, and close with light energy Degree enhancing, compound F-3 phototoxicity are presented conspicuousness and improve.

Claims (9)

1. oxazine compound, the structure with general formula F:
In general formula F,
The A is selected from oxygen, sulphur, selenium and tellurium；
The R1、R2And R3It is each independently selected from the group of hydrogen, formula i- X, C1-12Alkyl and C1-12Substitution alkyl；
The substitution alkyl is arbitrarily replaced by following radicals: halogen ,-OH ,-COOH ,-NO2、-SO3H、-OCH3、-OC2H5、- OC3H7、-OC4H9、-COOCH3、-COOC2H5、-COOC3H7、-COOC4H9、-COOC5H11、-COOC6H13、-NR4R5Or- CONR6R7；R therein4、R5、R6、R7It is each independently selected from hydrogen and C1-6Alkyl；
The X is selected from dihydrogen phosphate, bisulfate ion, nitrate anion, chlorine anion, bromine anion, iodine anion or perchloric acid Root.
2. oxazine compound according to claim 1, which is characterized in that the R1And R2Wherein at least one is hydrogen.
3. oxazine compound according to claim 1, which is characterized in that the R3It is hydrogen.
4. oxazine compound according to claim 1, which is characterized in that the R1、R2、R3It is each independently selected from Hydrogen, the group of formula i, formula ii group, C1-8Alkyl and C1-8Substitution alkyl.
5. oxazine compound according to claim 4, which is characterized in that the R1、R2、R3It is each independently selected from Hydrogen, the group of formula i, formula ii group, C1-6Alkyl and C1-6Substitution alkyl.
6. oxazine compound according to claim 1, which is characterized in that the R4、R5、R6、R7It is hydrogen.
7. oxazine compound according to claim 1, which is characterized in that it is selected from F-1, F-2, F-3, F-4, F-5, F- 6, F-7, F-8, F-9, F-10, F-11 or F-12:
8. oxazine compound described in claim 1 is preparing the application near infrared fluorescent probe, the near-infrared is glimmering Light probe is Mitochondrially targeted fluorescence probe.
9. oxazine compound described in claim 1 is preparing the application in light/sound sensitiser.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2021528482A (en) * 2018-05-04 2021-10-21 大連理工大学 Oxazine compounds and their use

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015066296A1 (en) * 2013-10-31 2015-05-07 Beth Israel Deaconess Medical Center Near-infrared fluorescent nerve contrast agents and methods of use thereof
CN106634964A (en) * 2016-11-09 2017-05-10 大连理工大学 Application of oxazine compound in preparation of near infrared fluorescence probe
CN107375929A (en) * 2017-08-04 2017-11-24 大连理工大学 A kind of sensitising agent and its derivative and application
CN108774249A (en) * 2018-05-04 2018-11-09 大连理工大学 Oxazine compounds and its application

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015066296A1 (en) * 2013-10-31 2015-05-07 Beth Israel Deaconess Medical Center Near-infrared fluorescent nerve contrast agents and methods of use thereof
CN106634964A (en) * 2016-11-09 2017-05-10 大连理工大学 Application of oxazine compound in preparation of near infrared fluorescence probe
CN107375929A (en) * 2017-08-04 2017-11-24 大连理工大学 A kind of sensitising agent and its derivative and application
CN108774249A (en) * 2018-05-04 2018-11-09 大连理工大学 Oxazine compounds and its application

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
B. RAMA RAJU, ET AL.: "Synthesis and Photophysics of New Benzophenoxazine Derivatives Fused with Julolidine", 《THE 18TH INTERNATIONAL ELECTRONIC CONFERENCE ON SYNTHETIC ORGANIC CHEMISTRY》 *
姜娜: "线粒体荧光探针最新研究进展", 《化工学报》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2021528482A (en) * 2018-05-04 2021-10-21 大連理工大学 Oxazine compounds and their use
JP7117453B2 (en) 2018-05-04 2022-08-12 大連理工大学 Oxazine-based compound and use thereof

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