CN116836565A - Water-soluble squaraine dye, and synthetic method and application thereof - Google Patents

Water-soluble squaraine dye, and synthetic method and application thereof Download PDF

Info

Publication number
CN116836565A
CN116836565A CN202310846412.6A CN202310846412A CN116836565A CN 116836565 A CN116836565 A CN 116836565A CN 202310846412 A CN202310846412 A CN 202310846412A CN 116836565 A CN116836565 A CN 116836565A
Authority
CN
China
Prior art keywords
water
compound
soluble
squaraine dye
dye
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202310846412.6A
Other languages
Chinese (zh)
Inventor
杜健军
刘圆
李鑫
潘静巍
樊江莉
彭孝军
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Dalian University of Technology
Ningbo Research Institute of Dalian University of Technology
Original Assignee
Dalian University of Technology
Ningbo Research Institute of Dalian University of Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Dalian University of Technology, Ningbo Research Institute of Dalian University of Technology filed Critical Dalian University of Technology
Priority to CN202310846412.6A priority Critical patent/CN116836565A/en
Publication of CN116836565A publication Critical patent/CN116836565A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B57/00—Other synthetic dyes of known constitution
    • C09B57/007—Squaraine dyes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/10—Non-macromolecular compounds
    • C09K2211/1018—Heterocyclic compounds
    • C09K2211/1025—Heterocyclic compounds characterised by ligands
    • C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Materials Engineering (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Biomedical Technology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The invention discloses a water-soluble squaraine dye, a synthesis method and application thereof. One type of water-soluble squaraine dye has the following structural general formula:the invention provides a water-soluble squaraine dye, a synthetic method and application thereof, wherein an amphiphilic squaraine dye is synthesized through design, and the squaraine dye is dyed in rigid azaindole squaraineHydrophilic diglycolamine is introduced into the material to regulate the hydrophilic and hydrophobic properties, so that the diglycolamine can be accurately positioned on cell membranes. Meanwhile, the change of aggregation form further improves the photo-thermal performance in organisms, and realizes real-time biological imaging and treatment.

Description

Water-soluble squaraine dye, and synthetic method and application thereof
Technical Field
The invention relates to the technical field of organic dyes, in particular to a water-soluble squaraine dye, a synthesis method and application thereof.
Background
Cell membranes are elastic semipermeable membranes consisting of phospholipid bilayers at the periphery of cells, and are particularly important in regulating substance exchange and maintaining the stability of the intracellular environment due to their selectivity and permeability, and abnormalities in cell membranes are also commonly used as a reference indicator for some related diseases. Fluorescence imaging, which is a high-sensitivity and high-resolution technique, has been widely used in cell membrane imaging, and plays an important role in cell membrane real-time imaging and detection.
Photothermal therapy is to convert an external light source into local heat energy by using a photothermal material, so that proteins, nucleic acids or other intracellular biomacromolecules are denatured, the permeability of cell membranes is affected, and finally irreversible damage is caused, and the photothermal therapy can be applied to ablation of microorganisms or cancers. As a minimally invasive local cancer treatment method, there is a great deal of attention in recent years from researchers, with the advantages of non-invasive and remote controllability.
However, currently studied cell membrane fluorescent dyes still have some drawbacks, such as low signal-to-noise ratio, high cytotoxicity, easy photo bleaching, etc., which prevent their application. In addition, there are few reports of dyes capable of targeting cell membranes for photothermal therapy. Therefore, a cell membrane targeting photothermal agent with high signal-to-noise ratio and good biocompatibility is synthesized, and is necessary for real-time imaging and treatment of cancers.
Disclosure of Invention
Aiming at the problems of low signal-to-noise ratio, higher cytotoxicity, easiness in photo-bleaching and the like of the conventional cell membrane fluorescent dye, the invention aims to provide a water-soluble squaraine dye, a synthetic method and application thereof. Meanwhile, the change of aggregation form further improves the photo-thermal performance in organisms, and realizes real-time biological imaging and treatment.
In order to achieve the above object, the technical scheme of the present invention is as follows: a water-soluble squaraine dye, which has a structure shown in a general formula I:
in the general formula I, the components are shown in the specification,
R 1 and R is 4 Each independently selected from at least one of carboxyalkyl, hydroxy, alkyl sulfonate having 1 to 18 carbons;
R 2 and R is 3 Each independently selected from at least one of hydrogen, aryl, alkyl having 1 to 4 carbons.
Further, the R 1 And R is 4 Each independently selected from any one of the following structural groups:
further, the method comprises the steps of,
the R is 2 And R is 3 Each independently selected from any one of the following structural groups:
the synthesis method of the water-soluble squaraine dye comprises the following steps:
(1) Adding 7-azaindole J-1 substituted by carboxyl into methanol solution, slowly dropwise adding SOCl at 0 DEG C 2 After 3-5h of reaction, refluxing at 60-80 ℃ for 3-6h, then adding the compound with R 1 Substituted amine reacts for 3-5 hours at the temperature of 40-70 ℃ to obtain an intermediate J-2;
(2) Dissolving intermediate J-2 obtained in step (1) in a first organic solvent at 50-120deg.C, adding a solvent containing R 2 Reacting the substituted halogenated alkane to obtain an intermediate J-3;
(3) Dissolving the intermediate J-3 prepared in the step (2) in a second organic solvent at 70-120 ℃, adding squaric acid, condensing under the catalysis of a first organic base, and purifying to obtain an intermediate J-4;
(4) Dissolving intermediate J-4 obtained in step (3) in a third organic solvent at 100-120deg.C, adding a solvent containing R 3 、R 4 Substituted 5-amide-2, 3-trimethyl-3H-pyrrolo [2,3-b]And (3) carrying out condensation reaction on the pyridine quaternary ammonium salt S-1 under the catalysis of a second organic base to obtain the fluorescent dye.
Further, in the step (2), the first organic solvent is at least one selected from benzene, toluene, o-dichlorobenzene, acetonitrile and acetone.
Further, in the step (3), the second organic solvent is at least one selected from ethanol, acetic acid, acetic anhydride, DMF, n-butanol, trimethyl orthoformate, triethyl orthoformate;
the first organic base is selected from at least one of triethylamine, pyridine and DIPEA.
Further, in the step (4), the third organic solvent is at least one selected from ethanol, acetic acid, acetic anhydride, DMF, trimethyl orthoformate, triethyl orthoformate;
the second organic base is selected from at least one of triethylamine, pyridine and DIPEA.
Further, in step (2), a catalyst having R is added 1 The substituted amine reacts for 3-5 hours at the temperature of 40-70 ℃ and then is subjected to recrystallization and purification to obtain an intermediate J-3, wherein the solvent used for recrystallization is at least one selected from methanol, ethanol, ethyl acetate and diethyl ether.
The application of a water-soluble squaraine dye in the biological and medical fields.
Further, the method is applied to cell imaging and tumor photothermal treatment.
Further, the excitation wavelength is 600-950nm and the fluorescence detection wavelength is 650-1000nm when the fluorescent dye is applied.
In summary, the invention has the following beneficial effects:
1. according to the dye, due to the introduction of hydrophilic groups, the hydrophilic and hydrophobic properties of molecules are changed, the lipophilic structure in the dye matrix can enter cells, and hydrophilic flexible chains connected with two ends repel lipid layers, so that the dye is accurately positioned on cell membranes, and real-time cell imaging is realized.
2. The invention obviously enhances the water solubility of the dye by modifying the azaindole squaraine dye, solves the problem that the azaindole squaraine dye is easy to aggregate in aqueous solution, simultaneously leads dye molecules to generate molecular vibration more easily, improves the photo-thermal performance in organisms, shows good photo-thermal effects in vitro and cell experiments, has good biocompatibility, and can be applied to the aspects of tumor photo-thermal treatment and the like.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments or the description of the prior art will be briefly described below, it will be obvious that the drawings in the following description are some embodiments of the present invention, and that other drawings can be obtained according to these drawings without inventive effort to a person skilled in the art.
FIG. 1 is a high resolution mass spectrum of compound 1 prepared in example 1 of the present invention;
FIG. 2 is a graph showing normalized emission spectra of Compound 1, compound 2, compound 3 prepared in example 1 of the present invention in dichloromethane;
FIG. 3 is a graph showing MTT assay of Compound 1 prepared in example 1 of the present invention;
FIG. 4 is a graph showing MTT assay of Compound 2 prepared in example 2 of the present invention;
FIG. 5 is a graph showing MTT assay of Compound 3 prepared in example 3 of the present invention;
FIG. 6 is a confocal image of compound 1, compound 2, compound 3 and comparative examples in cells, prepared in examples of the present invention.
Detailed Description
The present invention will be described in further detail below.
Unless otherwise indicated, the terms used herein have the following meanings.
The term "MTT" as used herein refers to a method of detecting cell survival and growth.
Instruments and devices employed in the examples:
in the column chromatography process, 200-300 mesh column chromatography silica gel purchased from Qingdao Megao group Co., ltd., 100-200 mesh column chromatography silica gel and 20-40 mesh analytically pure quartz sand purchased from Tianda chemical reagent factory are adopted.
In the process of detecting the compound, a mass spectrometer adopts a Synta G2-Si HDMS high-resolution mass spectrometer of Waters company in the United states, and adopts a double-needle electrospray ion source to detect the positive and negative modes of the compound.
Dye absorption and emission spectra were measured using a Cary 60 UV visible spectrophotometer and a Cary Eclipse fluorescence spectrophotometer from Agilent corporation.
Cytotoxicity assays were measured using a Varioskan LUX Multimode Microplate Reader instrument from thermofsher, usa.
Cell uptake experiments were measured using a single photon confocal microscopy instrument from Olympus corporation, japan.
Cell membrane-targeted squaraine dyes of formula I are described in detail below in connection with the examples.
In the general formula I, the components are shown in the specification,
R 1 and R is 4 Each independently selected from any one of the following structural groups:
R 2 and R is 3 Each independently selected from any one of the following structural groups:
specific examples of the compounds represented by the general formula I are given below, but the present invention is not limited to these specific examples.
The synthesis mechanism of the compound shown in the general formula I is as follows:
the compounds of the present invention represented by the general formula I can be synthesized by the methods described below.
Examples
Example 1
Manufacture of R 1 And R is 4 Are all hydroxy, R 2 And R is 3 Compound 1 which is the same as methyl group
(1) Production of intermediate 1.1
Carboxyl-substituted 7-azaindole (1 g,4.90 mmol) was added to a solution of 20mL of methanol and 4mL of LSOCl was slowly added dropwise at 0deg.C 2 After 3h reaction, transfer to oil bath, raise the temperature to 60 ℃, reflux for 6h, then add 2.6mL diglycolamine, react for 3-5h at 50 ℃, concentrate to give intermediate 1.1 (0.42 g,1.44mmol, 29% yield).
HRMS-ESI:m/z calcd.M + for C 15 H 22 N 3 O 3 + ,292.1661；found,292.1658.
1 HNMR(400MHz,Chloroform-d)δ8.79(d,J＝1.6Hz,1H),8.53(t,J＝6.8Hz,1H),8.30(d,J＝1.6Hz,1H),4.24(t,J＝7.4Hz,1H),3.70–3.51(m,8H),2.66(s,3H),1.36(s,6H).
(2) Production of intermediate 1.2
Intermediate 1.1 obtained in step (1) was dissolved in 20mL of acetonitrile at 65℃and reacted with methyl iodide (0.82 g,5.77 mmol) to give intermediate 1.2 (0.34 g,1.11mmol, yield 77%) by recrystallization purification.
HRMS-ESI:m/z calcd.M + for C 16 H 24 N 3 O 3 + ,306.1812；found,306.1813.
(3) Production of Compound 1
The intermediate 1.2 obtained in the step (2) was dissolved in 10mL of n-butanol at 115℃and added with squaric acid (0.07 g,0.57 mmol) for condensation reaction, and the resultant was purified by column chromatography to obtain compound 1 (0.025 g,0.04mmol, yield 6%).
HRMS-ESI:m/z calcd.M + for C 36 H 45 N 6 O 8 + ,689.3283；found,689.3299.
1 HNMR (400 mhz, chloro-d) delta 8.86 (d, j=1.4 hz, 1H), 8.74 (d, j=1.4 hz, 1H), 8.37-8.26 (m, 2H), 8.12-8.02 (m, 2H), 7.98 (d, j=1.4 hz, 1H), 7.24 (s, 1H), 4.08-3.98 (m, 5H), 3.68-3.50 (m, 16H), 3.39 (s, 3H), 1.52 (s, 6H), 1.41 (s, 6H) (see fig. 1).
Example 2
Manufacture of R 1 And R is 4 The same groups are carboxyl groups, R 2 And R is 3 Compound 2 which is the same as methyl group
(1) Production of intermediate 2.1
Carboxyl-substituted 7-azaindole (1 g,4.90 mmol) was added to a solution of 20mL of methanol and slowly dropped at 0deg.C with 4mL of SOCl 2 Reverse, oppositeAfter 5h, the reaction mixture was transferred to an oil bath, heated to 60℃and refluxed for 6h, followed by the addition of 2.8mL of amino-monoethylene glycol-carboxylic acid, reacted at 40℃for 5h, and concentrated to give intermediate 2.1 (0.51 g,1.60mmol, 33% yield).
HRMS-ESI:m/z calcd.M + for C 16 H 22 N 3 O 4 + ,320.1610；found,320.1601.
1 HNMR(400MHz,Chloroform-d)δ8.79(d,J＝1.6Hz,1H),8.64(td,J＝6.6,0.9Hz,1H),8.30(d,J＝1.4Hz,1H),3.76(t,J＝7.1Hz,2H),3.66–3.50(m,4H),2.63(s,3H),2.49(t,J＝7.0Hz,2H),1.36(s,6H).
(2) Production of intermediate 2.2
Intermediate 2.1 was dissolved in 20mL of acetonitrile at 65℃and reacted with methyl iodide (0.85 g,6.00 mmol) to give intermediate 2.2 (0.36 g,1.08mmol, 67% yield) by recrystallization purification.
HRMS-ESI:m/z calcd.M + for C 17 H 24 N 3 O 4 + ,334.1761；found,334.1750.
(3) Production of Compound 2
Intermediate 2 obtained in step (2) was dissolved in 10mL of n-butanol at 115℃and added with squaric acid (0.07 g,0.57 mmol) for condensation reaction, followed by purification by column chromatography to give compound 2 (0.028 g,0.04mmol, yield 7%).
1 HNMR(400MHz,Chloroform-d)δ8.86(d,J＝1.4Hz,1H),8.40(d,J＝1.6Hz,1H),8.37–8.30(m,1H),8.27(s,1H),8.10–8.02(m,2H),7.98(d,J＝1.4Hz,1H),7.24(s,1H),4.06(s,3H),3.76(t,J＝7.1Hz,4H),3.66–3.51(m,8H),3.39(s,3H),2.50(t,J＝7.1Hz,4H),1.60(s,6H),1.42(s,6H).
Example 3
Manufacture of R 1 And R is 4 Are all the same as alkyl sulfonate, R 2 And R is 3 Compound 1 which is the same as methyl group
(1) Production of intermediate 3.1
Carboxyl-substituted 7-azaindole (1 g,4.90 mmol) was added to a solution of 20mL of methanol and slowly dropped at 0deg.C with 4mL of SOCl 2 After 5h reaction, transfer to oil bath, raise the temperature to 60 ℃, reflux for 6h, then add 2.4ml of 3-aminopropanesulfonic acid, react for 3-5h at 40 ℃, concentrate to give intermediate 3.1 (0.58 g,1.64mmol, 33% yield).
HRMS-ESI:m/z calcd.M - for C 15 H 20 N 3 O 5 S - ,354.1129；found,354.1140.
1 HNMR(400MHz,Chloroform-d)δ8.79(d,J＝1.4Hz,1H),8.62(t,J＝6.8Hz,1H),8.30(d,J＝1.4Hz,1H),3.69(t,J＝7.1Hz,2H),3.65–3.59(m,2H),3.59–3.50(m,2H),3.35(t,J＝7.1Hz,2H),2.52(s,3H),1.37(s,6H) .
(2) Production of intermediate 3.2
Intermediate 3.1 obtained in step (1) was dissolved in 20mL of acetonitrile at 65℃and methyl iodide (0.92 g,6.56 mmol) was added for reaction, followed by purification by recrystallization to give intermediate 3.2 (0.43 g,1.16mmol, yield 71%).
HRMS-ESI:m/z calcd.M + for C 16 H 24 N 3 O 5 S + ,370.1437；found,370.1402.
(3) Production of Compound 3
The intermediate 3.2 obtained in the step (2) was dissolved in 10mL of n-butanol at 115℃and added with squaric acid (0.07 g,0.57 mmol) for condensation reaction, followed by purification by column chromatography to give compound 3 (0.027 g,0.03mmol, yield 6%).
1 HNMR(400MHz,Chloroform-d)δ8.86(d,J＝1.4Hz,1H),8.73(d,J＝1.6Hz,1H),8.34–8.26(m,1H),8.23(s,1H),8.10–8.02(m,2H),7.98(d,J＝1.4Hz,1H),7.24(s,1H),4.06(s,3H),3.70(t,J＝7.1Hz,4H),3.65–3.51(m,8H),3.41–3.31(m,7H),1.60(s,6H),1.42(s,6H).
Comparative example 1
Production of Compound 4
(1) Production of intermediate 4.1
7-azaindole (1.00 g,4 mmol) and iodomethane (1.77 g,8 mmol) were added to a 100mL two-necked round bottom flask containing 20mL of acetone and placed under nitrogen. The mixture was then heated to reflux overnight, the reaction was terminated, after cooling to room temperature, 50mL of diethyl ether was added to precipitate, and the resulting solid precipitate was filtered, washed with diethyl ether and dried to give intermediate 4.1 (1.64 g,5.4mmol, 87% yield) as a brown solid.
(2) Production of Compound 4
Under the protection of nitrogen, squaric acid (300 mg,2.6 mmol) and a compound 1.2 (1.59 g,5.3 mmol) are reacted in a mixed solvent of triethyl orthoformate and n-butanol under heating at 120 ℃, the reaction is stopped after stirring for 2 hours, the reaction liquid is dropped into 150mL of diethyl ether dropwise after cooling to room temperature, and the obtained crude product is recrystallized by a silica gel column, and then the crude product is purified by silica gel chromatography with 80:1 methylene chloride/methanol (v/v) as an eluting solvent to obtain a blue solid compound 4 (0.067 g,0.16mmol with the yield of 6%)
HRMS-ESI:m/z calcd.M + for C 26 H 26 N 4 O 2 + ,427.2129；found,427.2130.
1 HNMR(400MHz,Chloroform-d)δ8.25(d,J＝4.7Hz,2H),7.60(d,J＝7.2Hz,2H),7.05(dd,J＝7.3,5.1Hz,2H),6.07(s,2H),3.68(s,6H),1.80(d,J＝6.1Hz,12H).
Performance detection
The compounds prepared in examples 1-3 and comparative example 1 above were subjected to the following performance tests as follows:
test example 1
Fluorescence spectrometry of Compound 1, compound 2 and Compound 3 obtained in the above examples
Precisely weighing the dye subjected to vacuum drying by using a ten-thousandth balance, preparing 2mmol/L DMSO dye mother solution into a brown sample bottle, and storing in a refrigerator at 4 ℃ for later use.
When the ultraviolet-visible absorption spectrum and the fluorescence spectrum are tested, 3 mu L of dye mother solution is measured by a micropipette and is dissolved in a quartz cuvette containing 3mL of solvent to be tested, and the dye is uniformly mixed to obtain the dye with the concentration of 2.0 mu mol/L for testing the absorption spectrum and the fluorescence emission spectrum. All tests were completed at 25 ℃.
FIG. 2 is a graph of normalized emission spectra of Compound 1, compound 2, compound 3 in dichloromethane;
as shown in fig. 2, in the dichloromethane solution, the emission of the compound 1, the compound 2 and the compound 3 is between 640nm and 750nm, and in the near infrared region, the emission of the near infrared region is favorable for avoiding the influence of biological autofluorescence, and better resolution and presentation effect can be achieved, so that the dye is more favorable for the application of the dye in biological imaging and treatment.
Test example 2
Cytotoxicity test on Compound 1, compound 2 and Compound 3 prepared in examples
Toxicity of dye molecules to cells was assessed by MTT assay. The principle is as follows: succinate dehydrogenase in the mitochondria of living cells reduces exogenous MTT to water insoluble blue-violet crystalline Formazan (Formazan) and deposits in cells, whereas dead cells do not. Dimethyl sulfoxide (DMSO) can dissolve formazan in cells, and the light absorption value can be measured at 490nm and 570nm by using an enzyme-labeled instrument, so that the number of living cells can be indirectly reflected.
MCF-7 cells were seeded in 96-well plates and incubated for 24h (number of cells per well 1X 10 4 And (c) a). DMEM medium containing different concentrations of compound 1, compound 2, compound 3 was added to the wells, respectively, light/dark treatment was given, and cell activity was detected by MTT assay after 24h incubation. The experimental data are shown in fig. 3, 4 and 5.
As can be seen from fig. 3, 4, 5, compound 1, compound 2, compound 3 showed good biocompatibility in MCF-7 cells, which is advantageous for specific imaging of the dye in the cells. Meanwhile, compound 1, compound 2 and compound 3 with different concentrations are selected for phototoxicity experiments, and the results show that the dye has good phototoxicity effect, can effectively kill cells at 300mW, and prove that the dye can efficiently generate heat under illumination, so that the dye has excellent photo-thermal treatment effect, and can be applied to the fields of biological imaging and biological imaging.
Test example 3
Cell uptake experiments were performed on the compound 1, compound 2, compound 3 and compound 4 prepared in comparative example 1
MCF-7 cells were inoculated into confocal dishes one day in advance, after the cells were attached, washed three times with PBS solution, then 2mL of medium and 1. Mu.M of Compound 1, compound 2, compound 3, comparative example were added, and fluorescence images of the cells were taken with a Confocal Laser Scanning Microscope (CLSM) to observe the uptake of dye and cells. The excitation wavelength is 640nm, the transmitting and receiving wave bands are 650-700nm, and experimental data are shown in figure 6.
As can be seen from fig. 6, compound 1, compound 2, and compound 3 can specifically localize to the cell membrane within 2 hours, whereas comparative example compound 4 entered the cell and had strong fluorescence in the cell, but not localized to the cell membrane. Therefore, it was found that the water solubility of the compound 1, the compound 2 and the compound 3 was enhanced after the introduction of the water-soluble group, and the compound had an amphiphilic structure, so that the compound was more easily localized to the cell membrane. The lipophilic structure in the dye matrix enables the dye matrix to enter cells, and hydrophilic flexible chains connected at two ends are repelled from a lipid layer, so that the compound 1, the compound 2 and the compound 3 can be specifically positioned on cell membranes, and have the potential of ideal fluorescent biological probes.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.

Claims (10)

1. A water-soluble squaraine dye is characterized by having a structure of a general formula I:
in the general formula I, the components are shown in the specification,
R 1 and R is 4 May be the same or different, R 1 And R is 4 Each independently selected from at least one of carboxyalkyl, hydroxy, alkyl sulfonate having 1 to 18 carbons;
R 2 and R is 3 May be the same or different, R 2 And R is 3 Each independently selected from at least one of hydrogen, aryl, alkyl having 1 to 4 carbons.
2. A class of water-soluble squaraine dyes according to claim 1, wherein R 1 And R is 4 Identical, R 1 And R is 4 Each independently selected from any one of the following structural groups:
-COOH -OH
3. a water-soluble squaraine dye according to claim 1, characterized in that,
the R is 2 And R is 3 Identical, R 2 And R is 3 Each independently selected from any one of the following structural groups:
4. a method for synthesizing a water-soluble squaraine dye according to any one of claims 1 to 3, comprising the steps of:
(1) Adding 7-azaindole J-1 substituted by carboxyl into methanol solution, slowly dropwise adding SOCl at 0 DEG C 2 After 3-5h of reaction, refluxing at 60-80 ℃ for 3-6h, then adding the compound with R 1 Substituted amine reacts for 3-5 hours at the temperature of 40-70 ℃ to obtain an intermediate J-2;
(2) Dissolving intermediate J-2 obtained in step (1) in a first organic solvent at 50-120deg.C, adding a solvent containing R 2 Reacting the substituted halogenated alkane to obtain an intermediate J-3;
(3) Dissolving the intermediate J-3 prepared in the step (2) in a second organic solvent at 70-120 ℃, adding squaric acid, condensing under the catalysis of a first organic base, and purifying to obtain an intermediate J-4;
dissolving the intermediate J-4 in a third organic solvent at 100-120deg.C, adding a solvent containing R 3 、R 4 Substituted 5-amide-2, 3-trimethyl-3H-pyrrolo [2,3-b]And (3) carrying out condensation reaction on the pyridine quaternary ammonium salt S-1 under the catalysis of a second organic base to obtain the fluorescent dye.
5. The method for synthesizing a water-soluble squaraine dye according to claim 4, wherein in step (2), the first organic solvent is at least one selected from benzene, toluene, o-dichlorobenzene, acetonitrile and acetone.
6. The method for synthesizing a water-soluble squaraine dye according to claim 4, wherein in step (3), the second organic solvent is at least one selected from ethanol, acetic acid, acetic anhydride, DMF, n-butanol, trimethyl orthoformate, triethyl orthoformate;
the first organic base is selected from at least one of triethylamine, pyridine and DIPEA.
7. The method for synthesizing a water-soluble squaraine dye according to claim 4, wherein in step (3), the third organic solvent is at least one selected from ethanol, acetic acid, acetic anhydride, DMF, trimethyl orthoformate, and triethyl orthoformate;
the second organic base is selected from at least one of triethylamine, pyridine and DIPEA.
8. The method of claim 4, wherein R is added in the step (2) 1 The substituted amine reacts for 3-5 hours at the temperature of 40-70 ℃ and then is subjected to recrystallization and purification to obtain an intermediate J-3, wherein the solvent used for recrystallization is at least one selected from methanol, ethanol, ethyl acetate and diethyl ether.
9. Use of a class of water-soluble squaraine dyes according to any one of claims 1 to 3 in the biological and pharmaceutical fields.
10. The use of a class of water-soluble squaraine dyes according to claim 9, characterized in that the excitation wavelength is 600-950nm and the fluorescence detection wavelength is 650-1000nm.
CN202310846412.6A 2023-07-11 2023-07-11 Water-soluble squaraine dye, and synthetic method and application thereof Pending CN116836565A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202310846412.6A CN116836565A (en) 2023-07-11 2023-07-11 Water-soluble squaraine dye, and synthetic method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202310846412.6A CN116836565A (en) 2023-07-11 2023-07-11 Water-soluble squaraine dye, and synthetic method and application thereof

Publications (1)

Publication Number Publication Date
CN116836565A true CN116836565A (en) 2023-10-03

Family

ID=88172357

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202310846412.6A Pending CN116836565A (en) 2023-07-11 2023-07-11 Water-soluble squaraine dye, and synthetic method and application thereof

Country Status (1)

Country Link
CN (1) CN116836565A (en)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101544844A (en) * 2009-04-27 2009-09-30 东华大学 Water-soluble near infrared luminescent quinoline squaraine dye and preparation and application thereof
CN103030989A (en) * 2012-12-10 2013-04-10 北京化工大学 Synthesis method of water-soluble squarylium indocyanine multifunctional cell fluorescent dye
CN105238093A (en) * 2015-09-02 2016-01-13 北京化工大学 Amphiphilic indole squarylium cyanine dye and application thereof in long-acting marking of lysosome
CN113214672A (en) * 2021-04-13 2021-08-06 大连理工大学 Amide substituted azaindole-pentamethine cyanine dye, its synthesis method and application
CN114773875A (en) * 2022-03-15 2022-07-22 大连理工大学 Azaindole-squarylium cyanine dye, and synthesis method and application thereof

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101544844A (en) * 2009-04-27 2009-09-30 东华大学 Water-soluble near infrared luminescent quinoline squaraine dye and preparation and application thereof
CN103030989A (en) * 2012-12-10 2013-04-10 北京化工大学 Synthesis method of water-soluble squarylium indocyanine multifunctional cell fluorescent dye
CN105238093A (en) * 2015-09-02 2016-01-13 北京化工大学 Amphiphilic indole squarylium cyanine dye and application thereof in long-acting marking of lysosome
CN113214672A (en) * 2021-04-13 2021-08-06 大连理工大学 Amide substituted azaindole-pentamethine cyanine dye, its synthesis method and application
CN114773875A (en) * 2022-03-15 2022-07-22 大连理工大学 Azaindole-squarylium cyanine dye, and synthesis method and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PIECHOWSKI A. P.: "Desirable properties of photovoltaic dyes", JOURNAL OF PHYSICAL CHEMISTRY, vol. 88, no. 5, 31 December 1984 (1984-12-31), pages 934 - 950, XP002464705, DOI: 10.1021/j150649a020 *

Similar Documents

Publication Publication Date Title
CN105924394B (en) A kind of two-photon formaldehyde fluorescence probe and its preparation and application
Zhao et al. Maximizing the thiol-activated photodynamic and fluorescence imaging functionalities of theranostic reagents by modularization of BODIPY-based dyad triplet photosensitizers
Wang et al. Rational design of novel near-infrared fluorescent DCM derivatives and their application in bioimaging
CN110498799B (en) Fluorescent probe and preparation method and application thereof
CN114591632B (en) Azaindole-hemicyanine dye, and synthetic method and application thereof
WO2020221217A1 (en) Fluorescent dye, preparation method therefor, and use thereof
Wan et al. A ratiometric near-infrared fluorescent probe based on a novel reactive cyanine platform for mitochondrial pH detection
WO2013131235A1 (en) Two-photon fluorescent probe using naphthalene as matrix and preparation method and use thereof
WO2023092814A1 (en) Amino-substituted chromenoquinoline type fluorescent marker, and preparation and use thereof
CN108640902A (en) The fluorescence probe of sulfur dioxide and its application in a kind of identification pure aquatic system
Han et al. A diphenylacrylonitrile conjugated porphyrin with near-infrared emission by AIE–FRET
CN114773875B (en) Azaindole-squaraine dye, and synthetic method and application thereof
CN111334070B (en) 532nm excited rhodamine fluorescent dye and preparation method thereof
CN116836565A (en) Water-soluble squaraine dye, and synthetic method and application thereof
CN112010807B (en) Photosensitizer and application and preparation method thereof
CN115160345A (en) Azaindole-heptamethine cyanine dye, and synthesis method and application thereof
CN112939936B (en) Carbonyl azetidine substituted naphthalimide fluorescent dye and synthetic method and application thereof
CN114874639B (en) Novel hemicyanine near infrared fluorescent dye, and synthesis method and application thereof
CN116354883B (en) Compound for polarity detection and application thereof
CN115353460B (en) Application of phenol-containing keto-salicylaldehyde hydrazine compound in endoplasmic reticulum imaging
CN116120354A (en) Fluorescent start-type probe and preparation method and application thereof
CN115124555A (en) Perylene bisimide derivative-based dioxygenase fluorescent material and preparation method thereof
CN116854621A (en) Photosensitizer taking pentamethine cyanine dye as matrix, and synthesis method and application thereof
CN115124557A (en) FRET (fluorescence resonance energy transfer) hypoxic enzyme fluorescent probe based on perylene bisimide as well as preparation method and application thereof
CN117088881A (en) Azabicyclo-naphthalene fluorescent molecular probe and preparation and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination