CN117658973A - Infrared fluorescent dye and application thereof - Google Patents

Infrared fluorescent dye and application thereof Download PDF

Info

Publication number
CN117658973A
CN117658973A CN202311639934.5A CN202311639934A CN117658973A CN 117658973 A CN117658973 A CN 117658973A CN 202311639934 A CN202311639934 A CN 202311639934A CN 117658973 A CN117658973 A CN 117658973A
Authority
CN
China
Prior art keywords
compound
alkoxy
alkyl
substituted
group
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
CN202311639934.5A
Other languages
Chinese (zh)
Inventor
杨有军
钱旭红
鲁锡存
庄晓莉
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.)
East China University of Science and Technology
Original Assignee
East China University of Science and 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 East China University of Science and Technology filed Critical East China University of Science and Technology
Priority to CN202311639934.5A priority Critical patent/CN117658973A/en
Publication of CN117658973A publication Critical patent/CN117658973A/en
Pending legal-status Critical Current

Links

Landscapes

  • Plural Heterocyclic Compounds (AREA)

Abstract

The invention provides an infrared fluorescent dye and application thereof. Specifically, the present invention provides a compound represented by the following formula A, wherein R 1 ～R 17 And A ‑ Is defined herein. The dye molecules of the invention have maximum absorption wavelength distribution of 740nm-838nm, have large-range uniform distribution in the near infrared region, and show excellent fluorescence brightness and light stability, which are superior to other types of organic small molecule near infrared fluorescent dyes in the field.

Description

Infrared fluorescent dye and application thereof
Technical Field
The invention belongs to the field of dye chemistry, and particularly relates to an infrared fluorescent dye and application thereof.
Background
In the last twenty years, super-resolution fluorescence microscopy imaging technology has improved the resolution of dynamic microscopy imaging of living cells to the nanometer level. Among them, the development of fluorescent dyes for super-resolution microscopy imaging is the leading edge of research in the field of dye chemistry. Visible light super-resolution fluorescence microscopic imaging is interfered by biological background signals, has phototoxicity and is not suitable for long-time imaging of living biological samples. Near infrared dyes can break through the limitations and are of great interest in the field of super-resolution fluorescence microscopy imaging. The cyanine dye has high fluorescence brightness and small cytotoxicity, and can be widely applied to near infrared super-resolution fluorescence microscopic imaging. At the same time, however, cyanine dyes have disadvantages. At present, the cyanine dye applied to near infrared super-resolution fluorescence microscopic imaging has shorter wavelength, is distributed at 650-800nm and has poor light stability. However, the existing dyes with the wavelength of more than 800nm have the defects of light stability and fluorescence brightness. Therefore, development of super-resolution fluorescent microscopic imaging dye with wavelength larger than 800nm can realize long-time and low-toxicity dynamic fluorescent microscopic imaging, and can also develop multi-channel super-resolution imaging with visible light dye. In addition, the near infrared dye with the wavelength of more than 800nm can be used in the fields of in-vivo fluorescence imaging, photodynamic therapy of diseases, photothermal ablation of tumors and the like by utilizing the advantages of strong tissue penetrating capacity and low fluorescence background.
Disclosure of Invention
A first aspect of the present invention provides a compound of formula a:
wherein:
R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 and R is 8 Each independently selected from: hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- ，-SO 2 X，-SO 2 NH 2 And optionally 1 or 2 are selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 An alkyl group; or R is 1 And R is R 2 、R 2 And R is R 3 Or R 3 And R is R 4 And/or R 5 And R is R 6 、R 6 And R is R 7 Or R 7 And R is R 8 Together with the C to which they are each attached, form a group optionally consisting of 1 to 3 groups selected from halogen, C 1-4 Alkyl and C 1-4 Benzene ring substituted by substituent of alkoxy;
R 9 、R 10 、R 11 、R 12 、R 13 each independently selected from: hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- ，-SO 2 X，-SO 2 NH 2 Optionally 1 to 5 are selected from halogen, C 1-4 Alkoxy and azido substituents substituted C 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 Alkyl, optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene substituted aryl C 1-4 An alkoxy group;
R 14 and R is 15 Each independently selected from H, C optionally substituted with halogen 1-6 Alkyl, C 6-14 Aryl and C 2-4 Alkenyl groups; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a group which is optionally substituted with 1 to 5 groups selected from C 1-6 Alkyl, halogen and oxo substituted 3-8 membered heterocyclyl or 4-15 membered heterobridged heterocyclyl; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a 3-8 membered cycloalkyl spiro 4-15 membered heterobridged ring group or a 3-8 membered heterocyclyl spiro 4-15 membered heterobridged ring group; the heterocyclic group and the heterobridged ring group have 1 to 5A heteroatom selected from N, O and S;
R 16 and R is 17 Each independently selected from H, C optionally substituted with halogen 1-4 Alkyl, C 1-4 Alkoxy and C 2-4 Alkenyl groups; or R is 16 C, R to which it is attached 14 、R 14 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 5-7 membered nitrogen-containing heterocycle substituted by substituent of alkyl, and/or R 17 C, R to which it is attached 15 、R 15 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 A 5-7 membered nitrogen-containing heterocycle substituted with a substituent of an alkyl group;
A - acid radical ion for removing proton for acid, wherein the acid is one or more selected from HCl, trifluoroacetic acid, methanesulfonic acid, acetic acid and sulfuric acid;
x is halogen;
y is O or S;
n is an integer selected from 1-500.
The present invention also provides a fluorescent dye comprising a compound as described in any of the embodiments herein and optionally a solvent.
The present invention also provides a dye composition comprising a compound as described in any of the embodiments herein and optionally a solvent.
The invention also provides a kit comprising a compound as described in any of the embodiments herein and optionally a solvent, or a fluorescent dye or dye composition as described herein.
The invention also provides the use of a compound as described in any of the embodiments herein in the manufacture of a reagent for near infrared fluorescence imaging, photothermal therapy and/or photodynamic therapy.
Drawings
Fig. 1 is an absorption emission spectrum of compound 10 in methylene chloride.
Fig. 2 is an absorption emission spectrum of compound 12 in methylene chloride.
Fig. 3 is an absorption emission spectrum of compound 17 in dichloromethane.
Fig. 4 is an absorption emission spectrum of compound 21 in dichloromethane.
Fig. 5 is an absorption emission spectrum of compound 24 in methylene chloride.
Fig. 6 is an absorption emission spectrum of compound 26 in methylene chloride.
Fig. 7 is an absorption emission spectrum of compound 28 in methylene chloride.
Fig. 8 is an absorption emission spectrum of compound 30 in methylene chloride.
Fig. 9 is an absorption emission spectrum of compound 32 in dichloromethane.
Fig. 10 is an absorption emission spectrum of compound 34 in methylene chloride.
Fig. 11 is an absorption emission spectrum of compound 46 in methylene chloride.
Fig. 12 is a graph of super-resolution imaging results of compound 34 for nanospheres, wherein graph a is a contrast between wide-field imaging and structured light significant micro-imaging (SIM) super-resolution imaging of microspheres of different nanoparticle sizes, graph B is an imaging result of 300nm nanospheres, graph C is an SIM super-resolution imaging magnification of a box-selected region in graph B, and graph D is a wide-field fluorescence microscopy imaging magnification of a box-selected region in graph B.
Fig. 13 is a SIM super-resolution microscopic imaging and wide-field imaging of compound 36 for fixed cell (U2 OS cell line) mitochondria and living endocytosis.
FIG. 14 is a fluorescence imaging of mice injected with compounds 28, 24, 34 and DSPE-mPEG2000 nano-micelles after entrapment of compound 34.
FIG. 15 is a photothermal temperature increase curve of Compound 17. FIG. A is an aqueous solution of Compound 17 at 1W/cm at various concentrations 2 Temperature profile under 808nm laser irradiation of (c). Panel B is a plot of temperature over time for an aqueous solution of compound 17 at the same concentration (10. Mu.M) under 808nm laser irradiation at different power densities. FIG. C is a 15. Mu.M aqueous solution of Compound 17 at a power density of 1W/cm 2 A temperature rise curve (0 s-1200 s) and a temperature fall curve (1200 s-2400 s) under 808nm laser irradiation.
FIG. 16 is an ultraviolet absorbance spectrum generated using DPBF to detect singlet oxygen of a compound. Panel A shows a concentration of 20. Mu.MDPBF in acetonitrile 1W/cm 2 The ultraviolet absorption spectrum after the 808nm laser irradiation. Fig. B is an ultraviolet absorption spectrum of DPBF and compound 21 in acetonitrile solution and an ultraviolet absorption spectrum after mixing the two, respectively. FIG. C is a graph of a 1W/cm acetonitrile mixture of DPBF and compound 21 over various lengths of time 2 The ultraviolet absorption spectrum after the 808nm laser irradiation.
Detailed Description
It is understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described below (e.g., in the examples) may be combined with each other to constitute a preferred technical solution.
Terminology
The term "alkyl" as used herein, by itself or as part of another substituent, refers to a straight or branched hydrocarbon radical having the indicated number of carbon atoms (e.g., C 1 -C 20 、C 1 -C 10 Or C 1 -C 6 Wherein C 1 -C 20 Representing 1-20 carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.
As used herein, "alkenyl" refers to a straight or branched chain group containing 2 to 20 carbon atoms (e.g., 2 to 10 carbon atoms, 2 to 6 carbon atoms) unless the carbon chain length is otherwise limited, wherein at least two carbon atoms in the chain contain a double bond between them. Typical alkenyl groups include vinyl, 1-propenyl, 2-methyl-1-propenyl, 1-butenyl and 2-butenyl.
As used herein, "alkynyl" refers to a straight or branched chain group containing 2 to 20 carbon atoms (e.g., 2 to 10 carbon atoms, 2 to 6 carbon atoms) unless the carbon chain length is otherwise limited, wherein at least two carbon atoms in the chain contain a triple bond between them. Typical alkynyl groups include ethynyl, 1-propynyl, 1-methyl-2-propynyl, 1-butynyl and 2-butynyl.
As used herein, "halogen" or "halogen atom" refers to F, cl, br, and I. "halogenated" means selected from F, cl,Atoms of Br, and I. "halogen anion" refers to anions wherein the halogen atom is rendered one electron, including Cl — 、Br — 、I — 、F — 。
As used herein, "alkylene" refers to the same group as alkyl but with divalent groups. C (C) 1-4 Alkylene is a divalent group having 1 to 4 carbon atoms. Examples of alkylene groups include methylene (-CH) 2 (-), ethylene (-CH) 2 CH 2 (-), propylene (-CH) 2 CH 2 CH 2 -) and butylene (-CH) 2 CH 2 CH 2 CH 2 (-), etc.
As used herein, "heterocyclyl" refers to a saturated or partially saturated 3-7 membered monocyclic group or 7-10 membered bicyclic group consisting of carbon atoms and 1-4 heteroatoms selected from O, N and S. Examples of heterocyclyl groups include tetrahydrofuranyl, pyranyl, piperidinyl, piperazinyl, tetrahydro-1, 4-thiazinyl, tetrahydro-1, 3-thiazinyl, tetrahydro-1, 2-thiazinyl, 1, 3-dioxolanyl, 1, 2-oxazinyl, 1, 3-oxazinyl, 1, 4-diazinoheptyl, azetidinyl, oxetanyl, pyrrolidinyl, imidazolidinyl, imidazolinyl, indolinyl, isoindolinyl, morpholinyl, pyrazolidinyl, pyrazolinyl, and tetrahydroisoquinolinyl. "Nitrogen-containing heterocycle" means that the "heterocyclyl" contains at least one nitrogen atom, and also includes carbon atoms and optionally an additional 1 to 4 heteroatoms selected from O, N and S, examples of "nitrogen-containing heterocycles" include, but are not limited to, piperidine, tetrahydrothiazine, oxazinane.
As used herein, "oxo" refers to substitution with a group "=o". "oxo-heterocyclyl" means that a carbon atom in the heterocyclyl or a heteroatom selected from N or S is substituted by one, two or three groups "=o". Examples of "oxo-heterocyclyl" include, but are not limited to, oxo-pyrrolidinyl, 3-oxo-1, 2, 4-triazolidinyl, 5-oxo-1, 2, 4-triazolidinyl, dioxopyrrolidinyl, oxo-morpholinyl, oxo-piperidinyl, oxo-piperazinyl, dioxothiomorpholinyl, such as 1, 1-dioxothiomorpholinyl.
As used herein, "heteroaryl" refers to groups containing 5 to 14, preferably 5 to 10, ring atoms and having 6, 10 or 14 electrons in common on the ring system. The ring atoms contained in the heteroaryl group are carbon atoms and 1-3 heteroatoms selected from O, N and S. In the present invention, preferred heteroaryl groups are N-atom containing heteroaryl groups, more preferably N-atom containing 5-or 6-membered heteroaryl groups. Examples of heteroaryl groups include: triazolyl, thienyl, furyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, indolyl, benzimidazolyl, pyrazolopyrimidinyl and the like.
As used herein, "alkoxy" refers to an R-O-group, wherein R is an alkyl group as described herein. Preferred alkoxy groups are C 1-4 Alkoxy groups such as methoxy, ethoxy, propoxy, and the like.
As used herein, "aryl" refers to a monocyclic, bicyclic or tricyclic aromatic group containing 6 to 14 carbon atoms, i.e., C 6-14 Aryl, preferably C 6-10 Aryl groups. Examples of aryl groups include phenyl, naphthyl, phenanthryl, anthracyl, indenyl, azulenyl, biphenyl, biphenylene and fluorenyl.
Aryl C 1-4 Alkoxy refers to C substituted with aryl groups as described herein 1-4 An alkoxy group. Exemplary aryl C 1-4 Alkoxy includes benzyloxy.
As used herein, "cycloalkyl" or "carbocyclyl" is a saturated cyclic hydrocarbon having 3 to 13 ring carbon atoms, which contains one ring such as cyclohexyl or multiple rings such as adamantyl. Cycloalkyl groups containing more than one ring may be fused, spiro or bridged or a combination thereof. Preferred cycloalkyl groups are saturated cyclic hydrocarbons having 3 to 8 ring carbon atoms ("C 3-8 Cycloalkyl "). In some embodiments, the cycloalkyl group has 4 to 6 ring carbon atoms ("C 4-6 Cycloalkyl "). In some embodiments, the cycloalkyl group has 5 to 7 ring carbon atoms ("C 5-7 Cycloalkyl "). Examples of cycloalkyl groups include adamantyl, decalyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like.
As used herein, a "heterobridged cyclic" isRefers to a structure formed by two or more ring structures sharing two non-adjacent ring atoms with each other and having 1-4 heteroatoms selected from N, S, O on the ring. The heterobridged ring may have 4 to 15 ring atoms, preferably a 6 to 12 membered heterobridged ring, more preferably a 7 to 11 membered heterobridged ring. Examples of hetero-bridged rings include, but are not limited to: a "heterobridged ring radical" is a radical in which the heterobridged ring is attached to another radical to lose one hydrogen atom.
As used herein, "heterocyclyl spiro heterobridged ring radical" refers to a ketal obtained by reacting a bridged heterocycle containing a carbonyl group with ethylene glycol, for example, by spiro attachment of a heterocyclyl group as described herein to a heterobridged ring radical as described herein.
Compounds of the invention
The present invention provides a compound of formula a:
wherein:
R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 and R is 8 Each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 And optionally 1 or 2 are selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 An alkyl group; or R is 1 And R is R 2 、R 2 And R is R 3 Or R is 3 And R is R 4 And/or R 5 And R is R 6 、R 6 And R is R 7 Or R is 7 And R is R 8 Together with the C to which they are each attached, form a group optionally consisting of 1 to 3 groups selected from halogen, C 1-4 Alkyl and C 1-4 Benzene ring substituted by substituent of alkoxy;
R 9 、R 10 、R 11 、R 12 、R 13 each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 Optionally 1 to 5 are selected from halogen, C 1-4 Alkoxy and azido substituents substituted C 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 Alkyl, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy or aryl C 1-4 An alkoxy group;
R 14 and R is 15 Each independently selected from H, C optionally substituted with halogen 1-6 Alkyl, C 6-14 Aryl and C 2-4 Alkenyl groups; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a group which is optionally substituted with 1 to 5 groups selected from C 1-6 Alkyl, halogen and oxo substituted 3-8 membered heterocyclyl or 4-15 membered heterobridged heterocyclyl; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a 3-8 membered cycloalkyl spiro 4-15 membered heterobridged ring group or a 3-8 membered heterocyclyl spiro 4-15 membered heterobridged ring group; the heterocyclyl and heterobridged ring radicals have 1 to 5 heteroatoms selected from N, O and S;
R 16 And R is 17 Each independently selected from H, C optionally substituted with halogen 1-4 Alkyl, C 1-4 Alkoxy and C 2-4 Alkenyl groups; or R is 16 C, R to which it is attached 14 、R 14 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 5-7 membered nitrogen-containing heterocycle substituted by substituent of alkyl, and/or R 17 C, R to which it is attached 15 、R 15 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 A 5-7 membered nitrogen-containing heterocycle substituted with a substituent of an alkyl group;
A - acid radical ion for removing proton for acid, wherein the acid is one or more selected from HCl, trifluoroacetic acid, methanesulfonic acid, acetic acid and sulfuric acid;
x is halogen;
y is O or S;
n is an integer of 1 to 500.
In some embodiments, X is Cl.
In some embodiments, Y is O.
In some embodiments, a - Is Cl - 、F 3 CCOO - Or CH (CH) 3 SO 3 - 。
Preferably, n is an integer selected from 1-400, for example an integer selected from 10-350, an integer selected from 20-300, an integer selected from 50-250, an integer selected from 80-200. Preferably, n is an integer selected from 100-150. In some embodiments, n is an integer selected from 1 to 50. In some embodiments, n is an integer from 1 to 20. In some embodiments, n is an integer from 1 to 10.
In some embodiments, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 And R is 8 Each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 And optionally 1 or 2 selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 Alkyl, X is as described in any embodiment herein. In some embodiments, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 And R is 8 Each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 1-4 Alkyl, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 X is as described in any embodiment herein. Preferably, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 And R is 8 Each independently selected from hydrogen, halogen, C 1-4 Alkyl, -SO 3- and-SO 2 X, X are as described in any embodiment herein.
In some embodiments, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 And R is 8 Each independently is hydrogen, -SO 3- 、C 1-4 An alkyl group. Preferably, R 1 、R 2 、R 4 、R 5 、R 7 And R is 8 Is hydrogen, R 3 Is H or-SO 3- ，R 6 Is H or-SO 3- . More preferably, R 3 And R is 6 The substituents are the same.
In some embodiments, R 9 、R 10 、R 11 、R 12 、R 13 Each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 C optionally substituted with 1-5 substituents selected from halogen and azido 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 Alkoxy, halogen and azido substituted C 1-4 Alkyl, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy, X, Y and n are as defined above. In some embodiments, R 9 、R 10 、R 11 、R 12 、R 13 Each independently selected from hydrogen, -SO 3- 、-SO 2 X、-SO 2 NH 2 C optionally substituted with 1-3 substituents selected from halogen and azido 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 Alkoxy, halogen and azido substituted C 1-4 Alkyl, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy, X,Y and n are as defined above. In some embodiments, R 9 、R 10 、R 11 、R 12 、R 13 Each independently selected from: hydrogen, -SO 3- C optionally substituted with 1-3 substituents selected from halogen and azido 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 Alkoxy and halogen substituents 1-4 Alkyl, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy, X, Y and n are as defined above.
In some embodiments, R 9 、R 10 、R 12 、R 13 Each independently selected from hydrogen, -SO 3- And optionally C 1-4 Alkoxy substituted C 1-4 An alkyl group; r is R 11 Selected from hydrogen, C optionally substituted by 1-3 substituents selected from halogen and azido 1-4 Alkoxy and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 An alkoxy group.
In some embodiments, the oxo-heterocyclic group is a 1, 2, or 3 oxo-heterocyclic group. Preferably, the oxo-heterocyclic group is a dioxo-heterocyclic group. In some embodiments, the oxo-heterocyclyl is 2, 5-dicarbonyl-azaheterocyclyl. In the present invention, the oxo-heterocyclic group is a 5-7 membered heterocyclic group having 1 to 3 hetero atoms selected from O, S and N, such as succinimidylIn some embodiments, the oxo-heterocyclyl is a dioxo 4-6 membered heterocyclyl.
In some embodiments, R 9 、R 12 Each independently is H, -SO 3- Or optionally by C 1-4 Alkoxy substituted C 1-4 An alkyl group. Preferably, R 9 、R 12 Each independently is H, -SO 3- Or optionally by C 1-2 Alkoxy substituted C 1-2 An alkyl group. R is R 10 、R 13 Is hydrogen.R 11 Is hydrogen, C substituted by halogen and azido 1-4 Alkoxy or succinimidyl-OC (=o) -C 1-2 alkylene-O- [ C 1-2 alkylene-O] n -C 1-2 Alkylene-substituted heteroaryl C 1-4 Alkoxy, n is as defined above.
In some embodiments, R 14 And R is 15 Each independently selected from H, C 1-6 Alkyl and C 6-14 An aryl group; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a group which is optionally substituted with 1 to 3 groups selected from C 1-6 Alkyl, halogen and oxo substituted 4-7 membered heterocyclyl or 5-12 membered heterobridged heterocyclyl; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a 4-7 membered heterocyclyl spiro 5-12 membered heterobridged ring group; the heterocyclyl and heterobridged ring radicals have 1 to 3 heteroatoms selected from N, O and S. In some embodiments, R 14 And R is 15 Each independently selected from H, C 1-6 Alkyl and C 6-10 An aryl group; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a group which is optionally substituted with 1 to 3 groups selected from C 1-6 Alkyl, halo and oxo substituted 4-6 membered heterocyclyl or 6-9 membered heterobridged heterocyclyl; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a 4-6 membered heterocyclyl spiro 6-9 membered heterobridged ring group; the heterocyclyl and heterobridged ring radicals have 1-2 heteroatoms selected from N, O and S.
In some embodiments, the heterocyclyl has 1 or 2 groups selected from oxo, fluoro, chloro, C 1-4 Substituents of alkyl groups.
In some embodiments, R 16 And R is 17 Each independently selected from H, C 1-4 Alkyl and C 1-4 An alkoxy group; or R is 16 C, R to which it is attached 14 、R 14 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 5-7 membered nitrogen-containing heterocycle substituted by substituent of alkyl, and/or R 17 C, R to which it is attached 15 、R 15 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 Of alkyl groupsA 5-7 membered nitrogen-containing heterocycle substituted with a substituent.
In some embodiments, R 16 And R is 17 Each independently selected from H and C 1-4 An alkoxy group; or R is 16 C, R to which it is attached 14 、R 14 The N groups bound together form a 5-to 7-membered nitrogen-containing heterocycle, and/or R 17 C, R to which it is attached 15 、R 15 The attached N's together form a 5-7 membered nitrogen containing heterocycle. In some embodiments, R 16 And R is 17 Is C 1-4 Alkoxy, R 16 C, R to which it is attached 14 、R 14 The N groups attached together form a 5-7 membered nitrogen containing heterocycle, and R 17 C, R to which it is attached 15 、R 15 The attached N's together form a 5-7 membered nitrogen containing heterocycle.
Preferably, the compound of formula a is selected from the following compounds:
the near infrared fluorescent dye has novel structure and simple synthesis. The near infrared fluorescent dye has the maximum absorption wavelength of 740nm to 850nm, the maximum fluorescence emission wavelength of 780nm to 875nm, and the fluorescence emission peak has a stronger tail peak in the range of 1000nm to 1200nm, has strong infrared absorption, fluorescence emission capability and photoacoustic effect, and can be used for fluorescence and photoacoustic imaging of cells, tissues or living organisms. These dyes may also be used for imaging-based disease diagnosis, photodynamic therapy of disease or photothermal ablation of tumors.
The compounds of formula a of the present invention may be prepared from the corresponding starting materials with reference to the preparation examples provided in the examples herein.
Near infrared fluorescent dye or dye composition
The invention also provides near infrared fluorescent dyes or dye compositions containing the compounds of formula A of the invention. The dye or dye composition may also contain a solvent. Exemplary solvents include, but are not limited to, one or more of MeOH, etOH, prOH, iPrOH, buOH, water, acetonitrile, acetone, DMF, DMSO, pyridine, DCM, chloroform, dichloroethane, benzene, toluene, para-xylene, chlorobenzene, nitrobenzene, 1, 4-dioxane, THF, ethyl acetate, acOH, and buffers. In some embodiments, the solvent may be selected from water, acetonitrile or a buffer. Preferably, the buffer may be a PBS buffer.
In some embodiments, the dye composition contains a nanoparticle and a compound as described in any of the embodiments herein. Preferably, the nanoparticle is a polystyrene nanoparticle. The dye composition may be prepared by first preparing an aqueous solution of the compound described in any of the embodiments herein, and then mixing and shaking the aqueous solution/suspension containing the nanomicrospheres.
Kit for detecting a substance in a sample
The invention also provides a kit for fluorescence imaging, which contains the compound of the formula A, or a near infrared fluorescent dye or dye composition.
Application of
The present invention provides the use of a compound of formula a of the invention in the preparation of a reagent for near infrared fluorescence imaging, photothermal therapy or photodynamic therapy. Preferably, near infrared fluorescence imaging is nanomaterial imaging or cellular level imaging.
In some embodiments, the nanomaterial is a nanoparticle. Cell level imaging can image living cells and their organelles as well as fixed cells and their organelles. The cells can be selected from one or more of U2OS, hela, hepG2, RAW264.7, 4T1, A549, PC12 and other cell strains. In some embodiments, the compounds of the invention are used to image mitochondria and/or endocytosis. The method of cell fixation is a method commonly used in the art, for example, using paraformaldehyde to fix cells.
The advantages of the invention include:
(1) The invention provides a novel near infrared fluorescent dye mother nucleus structure, which is expected to be widely applied to the field of fluorescent dyes.
(2) The dye molecules of the invention have maximum absorption wavelength distribution of 740nm-838nm, have large-range uniform distribution in the near infrared region, and show excellent fluorescence brightness and light stability, which are superior to other types of organic small molecule near infrared fluorescent dyes in the field.
(3) The dye molecule provided by the invention has good biocompatibility, can realize in-vitro and cell-level super-resolution fluorescence microscopic imaging of material nanoparticles, fixed cell mitochondria, living cell endocytosis and the like, and has important application potential in super-resolution fluorescence microscopic imaging technology.
The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise indicated.
Example 1: preparation of Compound 10
(1) Synthesis of Compound 2: 30.00g of Compound 1 (147.78 mmol,1 equiv.) are weighed into a 500mL round bottom flask, a magnetic stirrer is added, 18.34g of ethylene glycol (295.55 mmol,2 equiv.) and 1g of p-toluene sulfonic acid as catalyst are added, 200mL of toluene are added for dissolution, and water is separated off in time by refluxing for 5 hours using a water separator. After the water in the water separator is not increased any more, stopping heating, cooling, adding 7.38mL of triethyl orthoformate (44.33 mmol,1 equiv.) and 5g of anhydrous magnesium sulfate, heating and refluxing for 2 hours by using a spherical condenser tube, stopping heating after the reaction is detected to be complete by thin layer chromatography, cooling to room temperature, neutralizing p-toluenesulfonic acid by using saturated sodium bicarbonate, adding 100mL of water-soluble salt, separating by a separating funnel, extracting the water phase by using DCM, drying by using a proper amount of anhydrous magnesium sulfate, filtering by suction, spin-drying toluene under reduced pressure, and distilling under reduced pressure to obtain 35.78g of compound 2 as colorless liquid with the yield of 98%.
1 H NMR(400MHz,CDCl 3 )δ7.59(s,1H),7.31(dd,J＝5.2,2.3Hz,1H),7.05(s,1H),6.04(s,1H),4.21–3.99(m,4H).
(2) Synthesis of Compound 3: 10.00g of Compound 2 (40.48 mmol,1 equiv.) are weighed into a 250mL round bottom flask, 100mL of anhydrous tetrahydrofuran is added to dissolve, a magnetic stirrer is added, 19.43mL of n-butyllithium (2.5M, 1.2 equiv.) is added at-78deg.C, stirring is maintained at-78deg.C for 15 minutes, 3.76mL of anhydrous DMF (48.57 mmol,1.2 equiv.) is added, gradually warmed to room temperature, after completion of the reaction by thin layer chromatography detection, quenched with saturated ammonium chloride solution, separated from the solution by a separating funnel, the aqueous phase is extracted with DCM, 3g of anhydrous magnesium sulfate is dried, suction filtered, and the solvent is spun dry under reduced pressure. Column chromatography (PE/EA, 100/5, v/v) gave compound 3 as a viscous pale yellow liquid, 6.35g, 80% yield.
1 H NMR(400MHz,CDCl 3 )δ10.40(d,J＝2.5Hz,1H),7.71(dd,J＝8.6,5.2Hz,1H),7.62(dd,J＝8.8,2.8Hz,1H),7.31–7.25(m,1H),6.30(s,1H),4.18–4.07(m,4H).
(3) Synthesis of Compound 4: 10.00g of Compound 3 (50.97 mmol,1 equiv.) and 3.98g (25.49 mmol,0.5 equiv.) of 1, 4-cyclohexanedione monoethylene glycol ketal were weighed in a 100mL round bottom flask, a magnetic stirrer was added, 10mL of ethanol was measured and dissolved, 30% by mass of sodium hydroxide solution was slowly added dropwise at room temperature until a pale yellow solid precipitated, 50mL of water was added to dilute the reaction solution, stirring was carried out for 30min, after the thin layer chromatography detection found that the reaction of the starting material was complete, suction filtration was carried out, a suitable amount of ice-ethanol washing was carried out, and compound 4 was recrystallized from dichloromethane and petroleum ether to give 12.80g as a pale yellow solid in 98% yield.
1 H NMR(400MHz,CDCl 3 )δ8.05(s,2H),7.60(dd,J＝8.6,5.8Hz,2H),7.06(td,J＝8.4,2.6Hz,2H),6.93(dd,J＝9.4,2.5Hz,2H),5.84(s,2H),4.17–4.09(m,4H),4.06–3.99(m,4H),3.87(s,4H),2.88(d,J＝1.5Hz,4H).
(4) Synthesis of Compound 5: 4.15g of diphenyl ether (24.39 mmol,2.5 equiv.) are weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, the mixture is placed in an ice-water bath for precooling for 10min, 9.76mL of n-butyllithium (2.5M, 2.5 equiv.) are added under the protection of argon gas, and the mixture is reacted for 1 hour at 0 ℃. 5.00g of Compound 4 (9.76 mmol,1 equiv.) are weighed into a 250mL round bottom flask, 120mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, the mixture is placed in an ice-water bath for precooling for 10min, the previously prepared diphenylether lithium reagent is transferred into the reaction solution, the reaction is carried out at 0 ℃ for 2 h, the reaction is detected by thin layer chromatography, the raw materials are completely reacted, the mixture is quenched by saturated ammonium chloride solution, 50mL of dissolved solid salt is added, a separating funnel is used for separating, the aqueous phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is dried, suction filtration is carried out, and the solvent is dried under reduced pressure. Dissolving the product in 50mL of dichloromethane, adding a magnetic stirrer, slowly dropwise adding 1mL of methanesulfonic acid, reacting for 30min at 40 ℃, stopping heating, cooling, adding 10mL of water, continuing to react for 4 hours at 40 ℃, after detecting that the reaction of the raw materials is complete by thin layer chromatography, adding saturated sodium bicarbonate solution to neutralize the methanesulfonic acid, adding 50mL of water, separating by a separating funnel, extracting the water phase with DCM, drying by a proper amount of anhydrous sodium sulfate, filtering, and spin-drying the solvent under reduced pressure. Column chromatography (PE/DCM, 3/1, v/v) gave compound 5 as a pale yellow solid 1.50g in 31% yield.
1 H NMR(400MHz,CDCl 3 )δ9.06(s,2H),8.05(dd,J＝8.8,5.7Hz,2H),7.50(s,2H),7.30(ddd,J＝12.0,8.4,3.5Hz,4H),7.25–7.16(m,4H),6.82–6.75(m,2H),6.61(dd,J＝7.9,1.4Hz,2H).
(5) Synthesis of Compound 6: 1.0g of Compound 5 (2.01 mmol,1 equiv.) is weighed into a 100mL round bottom flask, 30mL of dimethyl sulfoxide is added for dissolution, 1.45g of 30% by mass sodium methoxide methanol solution (8.06 mmol,4 equiv.) is added, a magnetic stirrer is added, the reaction is carried out at 80 ℃ for 5 hours, the reaction of the raw materials is found to be complete by thin layer chromatography detection, methanol is dried under reduced pressure, the reaction solution is poured into ice water, solid precipitation is carried out, suction filtration is carried out to obtain yellow solid, and column chromatography (PE/DCM, 2/1, v/v) is carried out to obtain Compound 6 as yellow solid 0.86g, and the yield is 82.5%.
1 H NMR(400MHz,CDCl 3 )δ8.98(s,2H),7.93(d,J＝9.1Hz,2H),7.41(s,2H),7.30(d,J＝8.3Hz,2H),7.19(t,J＝7.7Hz,2H),7.12(dd,J＝9.0,1.8Hz,2H),6.88(s,2H),6.78(t,J＝7.5Hz,2H),6.65(d,J＝7.9Hz,2H),3.83(s,6H).
(6) Synthesis of Compound 7: 500mg of compound 6 is weighed into a 250mL round bottom flask, 100mL of 1, 2-dichloroethane is added for dissolution, the mixture is placed in an ice-water bath for precooling for 10min, 1mL of boron tribromide is added under the protection of argon gas, after the mixture reacts for 30min under the ice-water bath, a spherical condenser tube is used, heating reflux is carried out for 6 hours at 60 ℃, thin layer chromatography detection shows that the raw materials react completely, cooling is carried out to room temperature, 20mL of water is slowly added dropwise from the upper opening of the condenser tube for quenching reaction, a separating funnel separates liquid, an aqueous phase is extracted by EA, a proper amount of anhydrous sodium sulfate is dried, suction filtration and reduced pressure spin-drying of the solvent are carried out, and column chromatography (PE/EA/DCM, 10/2/1, v/v) is carried out to obtain compound 7 which is light yellow solid 435.2mg, and the yield is 92%.
1 H NMR(400MHz,CDCl 3 )δ8.98(s,2H),7.95(d,J＝8.9Hz,2H),7.34(s,2H),7.28(s,2H),7.17(dd,J＝11.1,4.2Hz,2H),7.13–7.08(m,2H),6.91(s,2H),6.76(t,J＝7.5Hz,2H),6.67–6.60(m,2H),5.50(s,2H).
(7) Synthesis of Compound 8: 500mg of Compound 7 (1.02 mmol,1 equiv.) is weighed into a 250mL round bottom flask, 100mL of anhydrous dichloromethane is added for dissolution, a magnetic stirrer is added, the mixture is placed in an ice-water bath for precooling for 10min, 0.8mL of trifluoromethanesulfonic anhydride and 0.8mL of pyridine are slowly added for reaction for 1 hour at 0 ℃, the ice-water bath is removed, the reaction is continued for 1 hour at room temperature, thin layer chromatography detection shows that the raw materials are completely reacted, 30mL of water is slowly added for quenching reaction, a separating funnel is used for separating liquid, the aqueous phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is added for drying, suction filtration and reduced pressure spin-drying of the solvent are carried out, and column chromatography (PE/DCM, 3/1, v/v) is carried out to obtain Compound 8 which is 683.6mg as white solid with the yield of 89%.
1 H NMR(400MHz,CDCl 3 )δ9.10(s,2H),8.16(d,J＝9.1Hz,2H),7.62(s,2H),7.57(d,J＝2.3Hz,2H),7.38(dd,J＝9.1,2.4Hz,2H),7.34(dd,J＝8.3,1.1Hz,2H),7.26–7.20(m,2H),6.83–6.78(m,2H),6.57(dd,J＝7.9,1.5Hz,2H).
(8) Synthesis of compound 9: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 95.81mg of benzophenone imine (528.65. Mu. Mol,8 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, poured into the anaerobic reaction flask, catalytic amounts of BINAP and palladium acetate are added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is applied at the grinding port, a spherical condenser tube is connected, and the mixture is heated to 115 ℃ under the protection of argon and refluxed for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM/TEA, 1/1/0.01, v/v/v) gave compound 9 as a yellow solid 28mg in 51.7% yield.
1 H NMR(400MHz,CDCl 3 )δ8.87(s,2H),7.77(d,J＝8.8Hz,2H),7.72(d,J＝7.6Hz,4H),7.48(dd,J＝9.7,5.1Hz,2H),7.39(t,J＝7.5Hz,4H),7.29(s,2H),7.24(s,2H),7.22–7.07(m,12H),6.95(s,2H),6.82(dd,J＝8.7,1.7Hz,2H),6.77(t,J＝7.4Hz,2H),6.62(d,J＝7.8Hz,2H).HRMS(ESI)m/z:Calcd for C 60 H 39 N 2 O 2 + [M+H] + ,819.3006,Found,819.3013.
(9) Synthesis of Compound 10: 104.42mg (610.52 mu mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.24mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 100mg (126.43. Mu. Mol,1 equiv.) of Compound 9 is weighed into a 50mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 9 for reaction at zero ℃ for 5min, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating liquid, an aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, the crude product is dissolved in methanol, 1mL of 3M HCl is added for reaction for 1min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating liquid, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, column chromatography is carried out for chromatographic purification (DCM/MeOH=25:1 to 20:1), and 37mg of green solid is obtained, and the yield is 54%.
Compound 10 was dissolved to make a 2.5 μm solution of dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 10 in methylene chloride is shown in fig. 1.
1 H NMR(400MHz,MeOD)δ7.67(dd,J＝7.5,1.2Hz,1H),7.64(s,2H),7.63–7.49(m,5H),7.35(dd,J＝8.3,1.2Hz,2H),7.32–7.25(m,2H),7.02(s,2H),6.99–6.93(m,2H),6.91(dd,J＝9.1,2.0Hz,2H),6.86(dd,J＝7.9,1.4Hz,1H),6.80(dd,J＝7.9,1.4Hz,1H),6.58(d,J＝1.9Hz,2H),2.30(s,3H). 13 C NMR(101MHz,MeOD)δ168.59,158.54,150.25,150.20,150.14,143.34,140.82,137.70,136.84,136.59,131.83,131.43,131.22,131.07,131.03,130.95,130.50,130.36,129.84,127.60,127.22,127.03,125.24,125.15,121.12,118.28,118.16,108.85,48.67,20.04.HRMS(ESI)m/z:Calcd for C 41 H 29 N 2 O + [M] + ,565.2274,Found,565.2278.
Example 2: preparation of Compound 12
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 11: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 43mg of dimethylamine hydrochloride (528.6. Mu. Mol,8 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, the mixture is poured into the anaerobic reaction flask, a catalytic amount of BINAP and palladium acetate is added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is coated at the grinding port, a spherical condenser tube is connected, and the mixture is heated to 115 ℃ under the protection of argon and heated for reflux for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Compound 11 was used directly in the next reaction without isolation and purification due to poor solubility.
(3) Synthesis of Compound 12: 78.22mg (457.31. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.18mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (91.46. Mu. Mol,1 equiv.) of Compound 11 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 11, the reaction is carried out for 5min at zero ℃, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, an aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtered, spin-dried, the crude product is dissolved in dichloromethane, 1mL of trifluoroacetic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtered, spin-dried, and column chromatography is carried out for purification (DCM/MeOH=25:1 to 20:1), thus obtaining 40mg of purple solid with a yield of 70%.
Compound 12 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 12 in methylene chloride is shown in fig. 2.
1 H NMR(600MHz,CDCl 3 )δ7.61(td,J＝7.6,1.0Hz,1H),7.59–7.47(m,6H),7.39(d,J＝7.2Hz,1H),7.31(d,J＝8.2Hz,2H),7.26–7.22(m,2H),7.08(s,2H),7.05(dd,J＝9.4,2.4Hz,2H),6.92–6.84(m,2H),6.77(dd,J＝7.9,1.4Hz,1H),6.74(dd,J＝7.9,1.4Hz,1H),6.52(d,J＝2.3Hz,2H),3.22(s,12H),2.26(s,3H)； 13 C NMR(151MHz,CDCl 3 )δ154.9,149.1,149.0,135.3,131.3,131.1,130.3,130.1,128.9,128.9,126.4,124.4,124.3,117.5,117.5,117.5,107.4,41.3,20.2；HRMS(ESI)m/z:Calcd.for C 45 H 37 N 2 O + [M] + ,621.2900；Found,621.2908.
Example 3: preparation of Compounds 13,14 and 15
(1) The procedure for the synthesis of compounds 2-8 is described in example 1 and the procedure for the synthesis of compound 11 is described in example 2.
(2) Synthesis of Compound 13: 120.53mg (457.31. Mu. Mol,5 equiv.) of 2-bromo-4- (3-chloropropoxy) -toluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.18mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and after 20min of reaction, the reaction is carried out, the temperature is raised to room temperature for 5min; 50mg (91.46. Mu. Mol,1 equiv.) of Compound 11 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared phenyllithium reagent is transferred into a tetrahydrofuran solution of Compound 11, the reaction is carried out for 5min at zero ℃, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, the crude product is dissolved in dichloromethane, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, column chromatography and chromatographic separation and purification (DCM/MeOH=25:1 to 20:1) are carried out, and purple solid 46mg is obtained, and the yield is 70%.
1 H NMR(600MHz,Chloroform-d)δ7.64(s,2H),7.60(d,J＝9.5Hz,2H),7.36–7.32(m,3H),7.31–7.28(m,1H),7.10(d,J＝8.0Hz,2H),7.06(d,J＝6.8Hz,4H),6.96–6.92(m,1H),6.92–6.89(m,1H),6.80(dd,J＝7.9,1.3Hz,1H),6.76(dd,J＝7.9,1.3Hz,1H),6.53(d,J＝2.0Hz,2H),4.35(t,J＝5.7Hz,2H),3.89(t,J＝6.3Hz,2H),3.57(q,J＝7.1Hz,8H),2.39(p,J＝6.0Hz,2H),2.25(s,3H),1.25(t,J＝7.1Hz,12H). 13 C NMR(151MHz,Chloroform-d)δ166.10,159.90,152.92,148.85,148.72,148.54,141.12,139.30,135.39,130.90,130.22,129.97,129.18,128.64,128.61,126.64,125.80,124.23,124.17,117.27,117.17,117.12,116.85,112.18,106.74,64.54,47.36,45.85,41.66,32.28,20.33,13.04.
(3) Synthesis of Compound 14: 100mg of Compound 13 (129.80. Mu. Mol,1.0 equiv.) was weighed into a 25mL round bottom flask, 98% concentrated sulfuric acid (5 mL) was added to the round bottom flask under ice bath, stirred for 8h,TLC monitored the reaction was complete. Neutralizing with saturated sodium bicarbonate solution, spin-drying the solvent under reduced pressure, redissolving with methanol, and filtering to remove insoluble salts. Three drops of 3M hydrochloric acid solution were added and the solution changed from yellow to purplish red. Purifying the compound with reverse silica gel column with eluent ratio of H 2 O mecn=5:1 (v/v) to give 95.0mg of a purple solid in 71% yield.
1 H NMR(400MHz,MeOD)δ7.96(s,0H),7.76(ddd,J＝8.7,4.3,2.1Hz,1H),7.69(s,1H),7.65(d,J＝9.5Hz,1H),7.46(d,J＝8.7Hz,1H),7.43–7.37(m,2H),7.18–7.11(m,2H),6.81(s,1H),4.48(t,J＝5.5Hz,2H),4.03(t,J＝6.3Hz,1H),3.58(q,J＝6.9Hz,4H),2.46–2.41(m,1H),2.39(s,1H),1.19(t,J＝7.0Hz,6H)； 13 C NMR(101MHz,CD 3 OD)δ165.4,153.2,149.5,149.4,147.7,141.2,141.1,138.9,136.8,135.3,134.9,131.1,130.3,129.8,129.5,128.2,128.2,128.0,128.0,126.5,126.4,125.8,125.8,117.1,117.0,106.8,78.1,62.9,45.3,19.0,11.7；HRMS(ES + )calcd for C 49 H 43 N 2 Na 2 O 7 S 2 [M] 2- ,503.1021；Found,503.0990.
(4) Synthesis of Compound 15: 100mg of compound 14 (92.89. Mu. Mol,1.0 equiv.) are weighed into a 50mL round bottom flask, 5mL of N, N-dimethylformamide is added to dissolve, 12.08mg of sodium azide (18.58. Mu. Mol,2.0 equiv.) is added, and catalytic equivalent of potassium iodide (1 mg) is heated to 70℃and stirred for 24h. TLC monitored the reaction was complete. The reaction solution is directly purified by a reverse silica gel column, and the proportion of the eluent is H 2 O mecn=4:1 (v/v) to give 66mg of a purple solid with a yield of 66%.
ESI-HRMS(m/z)[M] 2- :calcd.For C 52 H 47 N 5 O 11 S 3 2- 506.6223,found 506.6227.
Example 4: preparation of Compound 17
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 16: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 28mg of N-methylaniline (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, the mixture is poured into the anaerobic reaction flask, a catalytic amount of BINAP and palladium acetate is added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is applied at the grinding port, a spherical condenser tube is connected, and the mixture is heated to 115 ℃ under the protection of argon and refluxed for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM, 3/1, v/v) gave compound 16 as a yellow solid 31mg in 70% yield.
1 H NMR(600MHz,CDCl 3 )δ8.88(s,2H),7.77(d,J＝9.1Hz,2H),7.36–7.33(m,4H),7.27(d,J＝3.5Hz,4H),7.19–7.16(td,J＝8.4,1.2Hz,2H),7.15(m,4H),7.13(s,2H),7.05(dd,J＝9.1,2.4Hz,2H),6.84(d,J＝2.4Hz,2H),6.77(td,J＝7.2,0.6Hz,2H),6.70(dd,J＝8.4,1.8Hz,2H),3.33(s,6H). 13 C NMR(151MHz,CDCl 3 )δ183.8,149.2,148.7,148.0,148.0,138.1,131.3,130.2,129.7,129.5,128.37,127.7,126.3,126.0,125.0,124.6,123.6,120.4,116.6,108.9,40.6.
(3) Synthesis of Compound 17: 63.74mg (372.68. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.15mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (74.54. Mu. Mol,1 equiv.) of Compound 16 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 16, the reaction is carried out for 5min at zero ℃, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtered, spin-dried, the crude product is dissolved in methylene chloride, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtered, spin-dried, and column chromatography is used for chromatographic separation and purification (DCM/MeOH=25:1 to 20:1), and the purple solid is obtained with 42mg, and the yield is 75%.
Compound 17 was dissolved to make a 2.5 μm solution of dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 17 in methylene chloride is shown in fig. 3.
1 H NMR(400MHz,CDCl 3 )δ7.69–7.62(m,3H),7.60–7.53(m,2H),7.47(m,7H),7.38(d,J＝7.6Hz,1H),7.37(t,J＝7.4Hz,1H),7.33(dd,J＝8.3,1.3Hz,2H),7.31–7.27(m,2H),7.20–7.15(m,4H),7.11(s,2H),6.98–6.88(m,4H),6.76(dd,J＝7.9,1.4Hz,1H),6.72(dd,J＝7.9,1.4Hz,1H),6.63(d,J＝2.2Hz,2H),3.52(s,6H),2.28(s,3H)； 13 C NMR(101MHz,CDCl 3 )δ168.7,154.9,149.2,149.1,149.1,145.3,141.7,140.3,136.5,135.1,135.1,131.8,131.3,130.8,130.6,130.3,130.1,130.1,129.1,129.1,129.0,128.9,128.4,127.2,127.1,126.8,126.5,124.6,124.5,124.5,119.4,117.8,117.7,109.2,47.8,41.9,20.3；HRMS(ESI)m/z:Calcd.for C 55 H 41 N 2 O + [M] + ,745.3213；Found,745.3217.
Example 5: preparation of Compound 19
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 18: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 45mg of diphenylamine (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, the mixture is poured into the anaerobic reaction flask, a catalytic amount of BINAP and palladium acetate are added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is coated at the grinding port, a spherical condenser tube is connected, and the mixture is heated to 115 ℃ under the protection of argon and heated for reflux for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM, 1/1, v/v) gave compound 18 as a yellow solid, 33mg, 64% yield.
1 H NMR(400MHz,CDCl 3 )δ8.91(s,2H),7.84(d,J＝9.0Hz,2H),7.28(s,4H),7.25–7.17(m,8H),7.15–7.04(m,16H),7.00(s,2H),6.75(t,J＝7.4Hz,2H),6.64(d,J＝7.8Hz,2H). 13 C NMR(101MHz,CDCl 3 )δ149.05,148.00,147.86,146.96,137.61,131.06,130.54,129.96,129.51,128.92,128.31,127.77,127.72,126.73,125.47,124.04,123.97,123.54,116.81,116.65,46.03.HRMS(EI)m/z:Calcd.for C 58 H 38 N 2 O 2 [M] + ,794.2933,Found,794.2931.
(3) Synthesis of Compound 19: 53.79mg (314.48. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.13mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (62.90. Mu. Mol,1 equiv.) of Compound 18 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 18, the reaction is carried out for 5min at zero ℃, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, the crude product is dissolved in methylene chloride, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, and column chromatography and chromatographic separation purification (DCM/MeOH=25:1 to 20:1) are carried out, thus obtaining a purple solid with a yield of 41mg and a yield of 75%.
1 H NMR(400MHz,CDCl 3 )δ7.72(s,1H),7.64(s,1H),7.55(d,J＝27.5Hz,1H),7.47–7.43(m,1H),7.41(d,J＝7.6Hz,1H),7.34–7.29(m,1H),7.28(s,5H),7.20(d,J＝3.8Hz,1H),7.15(d,J＝7.6Hz,6H),7.09–6.98(m,7H),6.91(s,5H),6.76(s,1H),6.70(dd,J＝15.3,7.3Hz,4H),2.28(s,3H). 13 C NMR(151MHz,CDCl 3 )δ170.50,154.40,149.08,148.74,148.71,144.19,141.37,140.46,136.17,134.87,134.51,131.83,131.07,130.64,130.56,130.29,130.02,129.80,128.85,128.26,128.12,128.03,127.76,127.54,126.93,126.28,124.30,124.24,122.02,117.59,117.49,113.89,47.51,19.99.HRMS(ESI)m/z:Calcd.for C 65 H 45 N 2 O + [M] + ,869.3526；Found,869.3533.
Example 6: preparation of Compound 21
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 20: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 15mg of cyclobutylamine (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, the mixture is poured into the anaerobic reaction flask, a catalytic amount of BINAP and palladium acetate is added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is coated at the grinding port, a spherical condenser tube is connected, and the mixture is heated to 115 ℃ under the protection of argon and heated for reflux for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM, 1/1, v/v) gave compound 20 as a yellow solid 25mg in 66% yield.
1 H NMR(400MHz,CDCl 3 )δ8.88(s,2H),7.85(d,J＝8.8Hz,2H),7.26-7.24(m,2H),7.22-7.13(m,4H),6.76-6.73(m,4H),6.71-6.66(m,2H),6.32(s,2H),3.94(t,J＝7.2Hz,8H),2.40-2.33(m,4H)； 13 C NMR(101MHz,CDCl 3 )δ157.6,144.5,142.2,131.3,128.8,123.6,103.4,58.5,51.9,18.4,16.8；HRMS(ESI)m/z:Calcd.for C 40 H 30 N 2 O 2 [M+H] + ,571.2380；found,571.2385.
(3) Synthesis of Compound 21: 74.93mg (438.06. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.18mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (87.61. Mu. Mol,1 equiv.) of compound 20 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of compound 20 for reaction for 5min at zero ℃, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating liquid, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, the crude product is dissolved into methylene chloride, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating liquid, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, column chromatography chromatographic separation and purification (DCM/MeOH=25:1 to 20:1) are carried out, and the purple solid 50mg is obtained, and the yield is 88%.
Compound 21 was dissolved to make a 2.5 μm solution of dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 21 in methylene chloride is shown in fig. 4.
1 H NMR(600MHz,CDCl 3 )δ7.62(td,J＝7.2,1.2Hz,1H),7.53(m,4H),7.48(d,J＝9.6Hz,2H),7.40(d,J＝7.2Hz,1H),7.31(d,J＝8.4Hz,2H),7.28–7.26(m,1H),7.26–7.24(m,1H),7.04(s,1H),6.92(td,J＝7.2,1.2Hz,1H),6.90(td,J＝7.2,1.2Hz,1H),6.79(dd,J＝7.8,1.8Hz,1H),6.75(dd,J＝7.8,1.8Hz,1H),6.65(dd,J＝9.0,1.8Hz,2H),6.19(d,J＝1.2Hz,1H),4.22(t,J＝7.2Hz,8H),2.51(m,4H),2.28(s,3H)； 13 C NMR(151MHz,CDCl 3 )δ154.3,149.1,149.1,140.9,139.6,135.4,131.2,130.9,130.4,130.3,130.2,129.0,128.9,126.5,124.5,124.4,117.6,117.5,115.8,105.3,52.2,20.3,16.4；HRMS(ESI)m/z:Calcd.for C 47 H 37 N 2 O + [M] + ,645.2900；Found,645.2907.
Example 7: preparation of Compound 22
(1) The procedure for the synthesis of compounds 2-8 is described in example 1 and the procedure for the synthesis of compound 20 is described in example 6.
(2) Synthesis of Compound 22: 88.08mg (438.06. Mu. Mol,5 equiv.) of 2-methoxymethyl bromobenzene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.18mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and after 20min of reaction, the reaction is carried out for 5min at room temperature; 50mg (87.61. Mu. Mol,1 equiv.) of compound 20 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of compound 20 for reaction for 5min at zero ℃, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating liquid, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, the crude product is dissolved into methylene chloride, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating liquid, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, column chromatography chromatographic separation and purification (DCM/MeOH=25:1 to 20:1) are carried out, and 52mg of purple solid is obtained, and the yield is 88%.
1 H NMR(400MHz,CDCl 3 )δ7.78-7.74(m,2H),7.65(t,J＝7.3Hz,1H),7.54(s,2H),7.47(t,J＝8.0Hz,3H),7.34-7.23(m,4H),7.04(s,2H),6.96-6.88(m,2H),6.85-6.79(m,2H),6.66(dd,J 1 ＝9.0Hz,J 2 ＝1.9Hz,2H),6.20(s,2H),4.40(s,2H),4.23(t,J＝7.5Hz,8H),3.16(s,3H),2.55-2.48(m,4H)； 13 C NMR(101MHz,CDCl 3 )δ149.9,149.0,148.5,143.9,134.6,130.3,130.1,129.8,129.4,128.8,128.7,128.6,126.2,124.2,124.1,117.3,117.2,115.0,103.8,72.8,58.5,52.3,51.9,16.1；HRMS(ESI)m/z:[M+H] + calculated for C 48 H 39 N 2 O 2 + ,675.3006；found,675.3011.
Example 8: preparation of Compound 24
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 23: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 34mg of 3, 3-difluoroazetidine hydrochloride (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, poured into the anaerobic reaction flask, catalytic amounts of BINAP and palladium acetate are added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is applied at the grinding port, a bulb condenser is connected, and the mixture is heated to 115 ℃ under the protection of argon and heated for reflux for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM, 3/1, v/v) gave compound 24 as a yellow solid 30mg in 71% yield.
1 H NMR(400MHz,CDCl 3 )δ8.93(s,2H),7.92(d,J＝8.9Hz,2H),7.29(s,3H),7.26(d,J＝4.3Hz,2H),7.16(t,J＝7.6Hz,2H),6.76(t,J＝9.1Hz,4H),6.65(d,J＝7.8Hz,2H),6.45(s,2H),4.26(t,J＝11.7Hz,8H)； 13 C NMR(101MHz,CDCl 3 )δ184.0,149.5,149.2,148.6,137.8,131.7,131.4,129.6,129.5,129.2,128.2,126.4,126.3,124.0,117.0,116.1,105.9,63.7,63.5,63.2,46.4；HRMS(EI)m/z:Calcd.for C 40 H 26 F 4 N 2 O 2 [M] + ,642.1930；Found,642.1934.
(3) Synthesis of Compound 24: 66.54mg (389.01. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.16mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (77.80. Mu. Mol,1 equiv.) of Compound 23 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared lithium methylbenzene is transferred into a tetrahydrofuran solution of Compound 23, reaction is carried out for 5min at zero ℃, the reaction is carried out for 2min at room temperature, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtered, spin-dried, the crude product is dissolved in methylene chloride, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtered, spin-dried, and column chromatography is used for chromatographic purification (DCM/MeOH=25:1 to 20:1), brown solid 46mg is obtained, and the yield is 83%.
Compound 24 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 24 in methylene chloride is shown in fig. 5.
1 H NMR(600MHz,CDCl 3 )δ7.78(s,2H),7.67(t,J＝7.4Hz,3H),7.57(dd,J＝16.1,7.7Hz,2H),7.44(d,J＝7.2Hz,1H),7.34(d,J＝8.3Hz,2H),7.29(dd,J＝6.8,5.2Hz,2H),7.19(s,2H),6.91(dt,J＝12.0,7.2Hz,2H),6.77(d,J＝7.0Hz,2H),6.74–6.67(m,2H),6.37(s,2H),4.53(t,J＝10.8Hz,8H),2.28(s,3H)； 13 CNMR(151MHz,CDCl 3 )δ170.6,153.1,149.3,149.0,148.9,141.5,141.3,135.9,131.1,131.0,130.5,129.9,129.7,129.6,129.0,128.9,128.4,128.3,127.0,126.9,126.2,124.3,124.3,117.5,117.4,116.3,114.5,106.6,63.2(t,J＝28.69Hz),47.6,20.0；HRMS(ESI)m/z:Calcd.for C 47 H 33 F 4 N 2 O + [M] + ,717.2524；Found,717.2527.
Example 9: preparation of Compound 26
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 25: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 19mg of cyclopentylamine (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, the mixture is poured into the anaerobic reaction flask, a catalytic amount of BINAP and palladium acetate is added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is coated at the grinding port, a spherical condenser tube is connected, and the mixture is heated to 115 ℃ under the protection of argon and heated for reflux for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM, 1/1, v/v) gave compound 25 as an orange solid 28mg in 70% yield.
1 H NMR(400MHz,CDCl 3 )δ8.87(s,2H),7.85(d,J＝9.1Hz,2H),7.24(s,2H),7.18(s,2H),7.14(dd,J＝14.8,1.6Hz,2H),6.95(dd,J＝9.0,1.8Hz,2H),6.76(d,J＝7.8Hz,2H),6.73–6.70(m,2H),6.42(s,2H),3.33(t,J＝6.2Hz,8H),1.99(t,J＝6.2Hz,8H). 13 C NMR(151MHz,CDCl 3 )δ183.6,149.1,147.9,147.5,138.3,131.3,131.0,129.9,128.6,128.4,127.4,124.8,124.6,123.5,116.7,116.3,103.7,47.6,46.1,25.4.HRMS(ESI)m/z:Calcd.for C 42 H 34 N 2 O 2 [M+H] + ,599.2693；found,599.2698.
(3) Synthesis of Compound 26: 71.41mg (417.54. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.17mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (77.80. Mu. Mol,1 equiv.) of compound 25 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared lithium methylbenzene is transferred into tetrahydrofuran solution of compound 25, reaction is carried out for 5min at zero ℃, the reaction is carried out for 2min at room temperature, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, the crude product is dissolved in methylene chloride, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, column chromatography chromatographic separation and purification (DCM/MeOH=25:1 to 20:1) are carried out, and the purple solid is obtained with 41mg, and the yield is 72%.
Compound 26 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 26 in methylene chloride is shown in fig. 6.
1 H NMR(400MHz,CDCl 3 )δ7.64(t,J＝7.5Hz,1H),7.61–7.49(m,6H),7.43(d,J＝7.4Hz,1H),7.34(d,J＝8.2Hz,2H),7.31–7.25(m,3H),7.09(s,2H),6.96(s,2H),6.93–6.88(m,2H),6.82(d,J＝7.9Hz,1H),6.79(d,J＝7.9Hz,1H),6.45(s,2H),3.54(s,8H),2.30(s,3H),2.08(s,8H). 13 C NMR(151MHz,CDCl 3 )δ152.2,148.9,148.8,148.4,140.9,139.3,135.1,130.9,130.8,130.1,130.0,129.9,128.6,126.1,124.2,124.1,118.1,117.3,117.2,107.6,48.9,25.2,20.0.HRMS(ESI)m/z:Calcd.for C 49 H 41 N 2 O + [M] + ,673.3213；Found,673.3218.
Example 10: preparation of Compound 28
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 27: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 35mg of 7-azabicyclo [2, 1] heptane hydrochloride (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, the mixture is poured into the anaerobic reaction flask, a catalytic amount of BINAP and palladium acetate are added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is applied at the grinding port, a spherical condenser tube is connected, and the mixture is heated to 115 ℃ under the protection of argon and heated for reflux for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM, 1/1, v/v) gave compound 27 as an orange solid 31mg in 72% yield.
1 H NMR(400MHz,CDCl 3 )δ8.89(s,2H),7.85(d,J＝9.0Hz,2H),7.28(d,J＝8.2Hz,2H),7.24(s,2H),7.16(dd,J＝13.6,4.8Hz,4H),6.79(s,2H),6.76(d,J＝7.6Hz,2H),6.69(d,J＝7.8Hz,2H),4.23(s,4H),1.75(d,J＝7.3Hz,9H),1.42(t,J＝6.4Hz,8H)； 13 C NMR(101MHz,CDCl 3 )δ184.2,149.5,148.5,148.1,138.3,131.6,131.2,129.8,129.6,128.7,128.0,126.7,126.2,123.9,120.3,116.8,109.6,58.1,46.4,29.1.HRMS(ESI)m/z:Calcd.for C 46 H 38 N 2 O 2 [M+H] + ,651.3006；found,651.3018.
(3) Synthesis of Compound 28: 65.70mg (384.13 mu mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.15mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (76.83. Mu. Mol,1 equiv.) of Compound 27 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 27, reaction is carried out for 5min at zero ℃, the reaction is carried out for 2min at room temperature, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtered, spin-dried, the crude product is dissolved in methylene chloride, 1mL of methanesulfonic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by methylene chloride, anhydrous sodium sulfate is dried, suction filtered, spin-dried, and column chromatography is used for chromatographic purification (DCM/MeOH=25:1 to 20:1), thus obtaining purple solid 42mg with 75% yield.
Compound 28 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 28 in methylene chloride is shown in fig. 7.
1 H NMR(600MHz,CDCl 3 )δ7.64(ddd,J＝7.5,7.5,0.8Hz,1H),7.58–7.50(m,6H),7.40(d,J＝7.2Hz,1H),7.34(d,J＝8.3Hz,2H),7.31–7.26(m,2H),7.09(s,2H),7.02(dd,J＝9.2,1.7Hz,2H),6.95–6.91(m,1H),6.90(m,1H),6.79(dd,J＝7.9,1.4Hz,1H),6.75(dd,J＝7.9,1.4Hz,1H),6.67(d,J＝1.4Hz,2H),4.48(s,4H),2.28(s,3H),1.80(d,J＝7.6Hz,8H),1.60(d,J＝7.3Hz,8H)； 13 C NMR(151MHz,CDCl 3 )δ165.8,151.8,149.2,149.1,148.9,141.5,139.2,136.5,135.9,135.2,131.3,131.2,130.4,130.3,130.2,130.0,129.2,129.1,129.0,129.0,127.4,127.2,126.5,124.5,124.5,119.4,117.7,117.6,109.9,77.6,77.4,77.1,58.0,29.1,29.1,20.2；HRMS(ESI)m/z:Calcd.for C 53 H 45 N 2 O + [M] + ,725.3526；Found,725.3530.
Example 11: preparation of Compound 30
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 29: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 26mg of N-methylpiperazine (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, poured into the anaerobic reaction flask, catalytic amounts of BINAP and palladium acetate are added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is applied at the mill, a bulb condenser is connected, and the mixture is heated to 115℃under the protection of argon and refluxed for 12 hours. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent.
1 H NMR(400MHz,CDCl 3 )δ8.91(s,2H),7.88(d,J＝9.2Hz,2H),7.30(s,2H),7.28(d,J＝8.3Hz,2H),7.21(dd,J＝9.2,1.9Hz,2H),7.15(t,J＝7.6Hz,2H),6.79(s,2H),6.75(t,J＝7.5Hz,2H),6.66(d,J＝7.8Hz,2H),3.34–3.22(m,8H),2.57–2.49(m,8H),2.32(s,6H). 13 C NMR(151MHz,CDCl 3 )δ183.89,150.74,149.19,147.94,137.90,131.21,130.66,129.79,129.40,128.31,127.78,126.41,126.20,123.66,119.58,116.61,108.57,54.80,48.31,46.05.HRMS(ESI)m/z:Calcd.for C 44 H 42 N 4 O 2 2+ [M+2H] 2+ ,329.1648；Found,329.1667.
(3) Synthesis of Compound 30: 65.10mg (380.62. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.15mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (76.12. Mu. Mol,1 equiv.) of Compound 29 is weighed in a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 29, reaction is carried out for 5min at zero ℃, the reaction is carried out for 2min at room temperature, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtered, spin-dried, the crude product is dissolved in dichloromethane, 1mL of trifluoroacetic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtered, spin-dried, column chromatography is carried out for chromatographic separation and purification (DCM/MeOH=10:1 to 1), and the purple solid is obtained with 29mg and the yield of 52%.
Compound 30 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 30 in methylene chloride is shown in fig. 8.
1 H NMR(600MHz,CD 2 Cl 2 )δ7.86(s,2H),7.68(s,3H),7.63–7.54(m,2H),7.48(d,J＝6.5Hz,1H),7.36(d,J＝7.4Hz,2H),7.30(d,J＝10.4Hz,4H),7.19(s,2H),6.91(d,J＝7.7Hz,2H),6.84(s,2H),6.75(dd,J＝24.0,7.3Hz,2H),3.91(s,8H),3.34(s,8H),2.84(s,6H),2.29(s,3H). 13 C NMR(151MHz,CD 2 Cl 2 )δ172.3,161.3,154.1,149.4,149.1,149.0,141.8,141.5,136.4,135.6,134.5,131.9,130.9,130.4,129.9,129.9,129.7,128.9,128.9,128.2,128.1,127.7,127.1,126.0,124.1,118.2,117.6,117.5,109.1,52.6,47.7,44.4,43.2,19.9.HRMS(ESI)m/z:Calcd.for C 51 H 47 N 4 O + [M] + ,731.3744；Found,731.3749.
Example 12: preparation of Compound 32
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 31: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 36mg of thiomorpholine dioxide (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, poured into the anaerobic reaction flask, catalytic amounts of BINAP and palladium acetate are added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is applied at the mill, a bulb condenser is connected, and the mixture is heated to 115℃under argon protection and refluxed for 12 hours. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent.
1 H NMR(400MHz,CDCl 3 )δ8.94(s,2H),7.97(d,J＝9.1Hz,2H),7.33(s,2H),7.29(d,J＝8.1Hz,2H),7.22–7.16(m,4H),6.85(d,J＝2.0Hz,2H),6.78(dd,J＝11.0,3.9Hz,2H),6.64(dd,J＝7.9,1.3Hz,2H),4.03–3.85(m,8H),3.15–3.01(m,8H). 13 C NMR(151MHz,CDCl 3 )δ183.67,149.13,148.31,137.63,131.74,131.09,129.94,129.04,128.46,128.04,126.68,126.65,123.82,118.65,116.73,109.59,50.50,46.94,46.14.HRMS(ESI)m/z:Calcd.for C 42 H 34 N 2 NaO 6 S 2 + [M+Na] + ,749.1750；Found,749.1755.
(3) Synthesis of Compound 32: 58.83mg (343.94. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.14mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (68.79. Mu. Mol,1 equiv.) of Compound 31 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 31, the reaction is carried out for 5min at zero ℃, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, an aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtered, spin-dried, the crude product is dissolved in dichloromethane, 1mL of trifluoroacetic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtered, spin-dried, and column chromatography is used for chromatographic separation and purification (DCM/MeOH=20:1 to 10:1), and 24mg of purple solid is obtained, and the yield is 44%.
Compound 32 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 32 in methylene chloride is shown in fig. 9.
1 H NMR(600MHz,CDCl 3 )δ7.84(s,2H),7.74(s,2H),7.68(t,J＝7.1Hz,1H),7.61–7.56(m,2H),7.45(d,J＝6.9Hz,1H),7.37–7.26(m,8H),6.95–6.87(m,2H),6.85(s,2H),6.69(dd,J＝21.6,7.7Hz,2H),4.15(s,8H),3.11(s,8H),2.28(s,3H). 13 C NMR(151MHz,CDCl 3 )δ172.6,160.1,159.8,153.0,149.5,149.0,148.9,142.0,141.7,136.6,136.1,134.2,132.1,131.2,130.7,130.0,129.9,129.7,129.1,128.1,128.0,127.7,127.1,126.3,124.4,118.4,117.7,117.6,116.5,114.6,109.4,51.9,47.7,46.5,20.3.HRMS(ESI)m/z:Calcd.for C 49 H 41 N 2 O 5 S 2 + [M] + ,801.2451；Found,801.2458.
Example 13: preparation of Compound 34
(1) The synthetic procedure for compounds 2-8 is as described in example 1.
(2) Synthesis of Compound 33: 50mg of Compound 8 (66.08. Mu. Mol,1 equiv.) are weighed into a 50mL anaerobic reaction flask, a magnetic stirrer is added, argon is introduced for 5min, 45mg of nortropinone ethylene glycol ketal (264.3. Mu. Mol,4 equiv.) and 215.31mg of cesium carbonate (660.81. Mu. Mol,10 equiv.) are added, poured into the anaerobic reaction flask, catalytic amounts of BINAP and palladium acetate are added, 35mL of dry toluene is added for dissolution, argon is introduced for 10min, vacuum silicone grease is coated at the grinding port, a spherical condenser is connected, and the temperature is raised to 115 ℃ under the protection of argon and heated for reflux for 12 h. The thin layer chromatography detection shows that the raw materials are completely reacted, the temperature is reduced to room temperature, 20mL of water is added for stirring, a separating funnel is used for separating liquid, the water phase is extracted by DCM, a proper amount of anhydrous sodium sulfate is used for drying, suction filtration and decompression spin-drying are carried out on the solvent. Column chromatography (PE/DCM, 3/1, v/v) gave compound 33 as an orange solid 45mg in 85% yield.
1 H NMR(400MHz,CDCl 3 )δ8.87(s,2H),7.87(d,J＝9.1Hz,2H),7.25(d,J＝6.6Hz,3H),7.19(s,2H),7.15(t,J＝7.7Hz,2H),7.10(dd,J＝9.1,1.8Hz,2H),6.78(t,J＝7.5Hz,2H),6.71(d,J＝6.9Hz,2H),6.66(s,2H),4.31(s,4H),3.95(t,J＝6.3Hz,4H),3.72(t,J＝6.3Hz,4H),2.16(t,J＝6.4Hz,4H),2.02(dd,J＝13.9,2.6Hz,4H),1.98–1.90(m,4H),1.67(d,J＝13.7Hz,4H)； 13 C NMR(101MHz,CDCl 3 )δ184.1,149.4,148.3,146.2,138.8,131.7,130.0,129.2,128.7,127.9,125.8,125.7,124.1,118.7,116.7,108.0,107.9,64.9,63.3,54.0,46.4,37.9,27.7；HRMS(ESI)m/z:Calcd.for C 52 H 46 N 2 O 6 [M+H] + ,795.3429；Found,795.3438.
(3) Synthesis of Compound 34: 53.79mg (314.49. Mu. Mol,5 equiv.) of o-bromotoluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.14mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and the reaction is carried out for 20min and then the reaction is carried out for 5min at room temperature; 50mg (62.90. Mu. Mol,1 equiv.) of Compound 33 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added for dissolution, the prepared methylbenzyllithium is transferred into a tetrahydrofuran solution of Compound 33 for reaction at zero ℃ for 5min, the reaction is carried out at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating liquid, an aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, the crude product is dissolved in dichloromethane, 1mL of trifluoroacetic acid is added for reaction for 10min, saturated sodium bicarbonate is added for neutralization, the separating funnel is used for separating liquid, the aqueous phase is extracted three times by dichloromethane, anhydrous sodium sulfate is dried, suction filtration and spin drying are carried out, column chromatography chromatographic separation and purification (DCM/MeOH=20:1 to 10:1) are carried out, and 47mg of purple solid is obtained, and the yield is 85%.
Compound 34 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 34 in methylene chloride is shown in fig. 10.
1 H NMR(400MHz,CDCl 3 )δ7.67–7.62(m,1H),7.61(s,2H),7.58(s,2H),7.54(d,J＝7.3Hz,1H),7.42(d,J＝7.1Hz,1H),7.36–7.27(m,4H),7.26–7.23(m,1H),7.16(d,J＝8.7Hz,2H),7.07(s,2H),6.92(ddd,J＝16.4,8.2,1.0Hz,2H),6.80(dd,J＝7.9,1.0Hz,1H),6.76(dd,J＝7.8,1.1Hz,1H),6.60(s,2H),4.55(s,4H),3.97(t,J＝6.3Hz,4H),3.75(t,J＝6.3Hz,4H),2.28(d,J＝5.0Hz,7H),2.03–1.84(m,12H)； 13 C NMR(151MHz,CDCl 3 )δ150.7,148.8,148.7,148.6,141.4,139.1,135.8,131.0,130.1,130.1,129.9,128.7,128.7,126.4,126.2,124.4,124.3,124.3,123.5,118.8,117.3,117.2,108.8,106.7,64.8,63.3,55.1,31.5,30.1,26.8,20.0；HRMS(ESI)m/z:Calcd.for C 59 H 53 N 2 O 5 + [M] + ,869.3949；Found,869.3953.
Example 14: preparation of Compounds 35 and 36
(1) The procedure for the synthesis of compounds 2-8 is described in example 1 and the procedure for the synthesis of compound 33 is described in example 13.
(2) Synthesis of Compound 35: 82.89mg (314.49. Mu. Mol,5 equiv.) of 2-bromo-4- (3-chloropropoxy) -toluene is weighed into a 100mL round bottom flask, 40mL of anhydrous tetrahydrofuran is added for dissolution, a magnetic stirrer is added, 0.14mL (2.5M, 5 equiv.) of n-butyllithium is added at-78 ℃, and after 20min of reaction, the reaction is carried out, the temperature is raised to room temperature for 5min; 50mg (62.90. Mu. Mol,1 equiv.) of compound 33 is weighed into a 100mL round bottom flask, 20mL of anhydrous tetrahydrofuran is added to dissolve, the prepared lithium methylbenzene is transferred into the tetrahydrofuran solution of compound 33, the reaction is carried out for 5min at zero ℃, the solution is placed at room temperature for 2min, TLC detects that the raw materials are completely reacted, saturated ammonium chloride solution is used for quenching the reaction, water is added, a separating funnel is used for separating, the aqueous phase is extracted three times by dichloromethane, and the solution is dried by anhydrous sodium sulfate, filtered by suction and dried by spin. The crude product was dissolved in 10ml of n, n-dimethylformamide, 8.18mg of sodium azide (125.79 μmol,2 equiv.) was added, catalytic equivalent of potassium iodide (1 mg) was added, and the mixture was heated to 70 ℃ for 2 hours. 50mL of water and 50mL of methylene chloride were added, and the organic phase was dried over anhydrous sodium sulfate, filtered off with suction, and dried with spin. The crude product was dissolved in 10mL of dichloromethane, 1mL of trifluoroacetic acid was added, reacted for 10min, neutralized with saturated sodium bicarbonate, separated by a separating funnel, the aqueous phase was extracted three times with dichloromethane, dried over anhydrous sodium sulfate, suction filtered, dried by spin-on, and purified by column chromatography (DCM/meoh=20:1 to 10:1) to give 42mg of purple solid with a yield of 69%.
1 H NMR(400MHz,CDCl 3 )δ7.65(s,2H),7.63(s,2H),7.32(dd,J＝9.2,4.8Hz,3H),7.29–7.26(m,1H),7.26–7.22(m,1H),7.16(d,J＝8.6Hz,2H),7.12–7.07(m,2H),7.06(s,2H),6.95–6.91(m,1H),6.89(dd,J＝9.6,2.7Hz,1H),6.78(dd,J＝7.9,1.1Hz,1H),6.73(dd,J＝7.9,1.2Hz,1H),6.59(s,2H),4.54(s,4H),4.29(t,J＝5.8Hz,2H),3.97(t,J＝6.3Hz,4H),3.74(t,J＝6.3Hz,4H),3.65(t,J＝6.5Hz,2H),2.28(d,J＝7.5Hz,4H),2.25(s,3H),2.23–2.18(m,2H),2.02–1.83(m,13H)； 13 C NMR(151MHz,CDCl 3 )δ166.8,160.2,150.9,149.2,149.0,148.9,141.7,139.7,138.4,136.1,131.5,131.2,130.5,130.2,129.4,129.3,129.0,128.9,127.6,127.3,126.7,124.6,124.5,119.0,117.5,117.4,117.2,112.5,109.0,107.1,65.2,65.1,63.6,55.3,48.6,47.7,41.0,31.9,29.2,27.1,23.0,20.7,14.4；HRMS(ESI)m/z:Calcd.for C 62 H 58 N 5 O 6 + [M] + ,968.4382；Found,968.4388.
(3) Synthesis of Compound 36: 20mg (20.64. Mu. Mol,1 equiv.) of compound 35 and 41.3mg Alkyne-PEG-NHS (20.64. Mu. Mol,1 equiv.) are weighed out and dissolved in 2mL anhydrous N, N-dimethylformamide in a 20mL round bottom flask. Argon is replaced for three times, and the bottle mouth is sealed by the rubber plug. 7.69mg of copper tetraacetonitrile hexafluorophosphate (20.64. Mu. Mol,1 equiv.) and 2.67mg of DIPEA (20.64. Mu. Mol,1 equiv.) were dissolved in 2mL of anhydrous N, N-dimethylformamide and added to the round bottom flask with a syringe. Stirred at room temperature for 2 hours. TLC showed complete reaction of starting materials, addition of water, separation of liquid by separating funnel, extraction of aqueous phase with dichloromethane three times, drying over anhydrous sodium sulfate, suction filtration, spin-drying, column chromatography purification (DCM/MeOH=20:1 to 10:1) gave 19mg of purple solid in 32% yield.
MALDI-TOF m/z Expected M.W.:～2968,Measured M.W.:～2988.
Example 15: preparation of Compound 46
(1) Synthesis of Compound 38: 20g of 2-nitrobenzene-1, 3-diol (129 mmol,1 equiv.) 54mL of 1-bromo-2-chloroethane (640 mmol,5 equiv.) and 10g of potassium carbonate are dissolved in 200mL of acetonitrile and heated to 90℃for 48 hours. Cooling to room temperature, and suction filtering. The filtrate was poured into 200mL petroleum ether and the solid precipitated. Suction filtration and drying gave 35g of product 38 as a white solid in 97% yield.
1 H NMR(400MHz,CDCl 3 )δ7.34(t,J＝8.0Hz,1H),6.67(d,J＝8.0Hz,2H),4.30(t,J＝4.0Hz,4H),3.78(t,J＝4.0Hz,4H)； 13 C NMR(101MHz,CDCl 3 )δ150.5,132.8,131.3,106.5,69.5,41.0；HRMS(ES + )calcd for C 10 H 11 Cl 2 NO 4 [M+Na] + ,301.9957；Found,301.9961.
(2) Synthesis of Compound 39: 10g of compound 38 (35.7 mmol,1 equiv.) are dissolved in 200mL ethyl acetate and stirred at room temperature for 24 hours. The reaction was quenched by addition of sodium hydroxide solution and extracted 3 times with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered off with suction and the solvent was dried under reduced pressure to give 11.7g of product 39 as a yellow oil in 91% yield.
1 H NMR(400MHz,CDCl 3 )δ7.60(d,J＝12.0Hz,1H),6.76(d,J＝8.0Hz,1H),4.36(t,J＝8.0Hz,2H),4.30(t,J＝8.0Hz,2H),3.79(t,J＝8.0Hz,2H),3.78(t,J＝8.0Hz,2H)； 13 C NMR(101MHz,CDCl 3 )δ149.7,148.1,137.9,134.8,111.0,108.8,74.4,69.9,41.6,40.9；HRMS(ES + )calcd for C 10 H 10 BrCl 2 NO 4 [M+Na] + ,379.9062；Found,379.9070.
(3) Synthesis of Compound 40: 20g of Compound 39 (55.76 mmol,1 equiv.) are dissolved in 100mL of ethanol, 2.2mL of acetic acid (39 mmol,0.7 equiv.) and 12.45g of iron powder (223 mmol,4 equiv.) are added, followed by 1mL of water. Heating to 90 ℃ under the protection of argon gas, and reacting for 12 hours. The reaction solution was suction filtered and the filtrate was adjusted to ph=12. The organic phase was dried over anhydrous sodium sulfate, filtered off with suction and dried. Column chromatography (PE/dcm=1/1, v/v) gave 12.4g of product 40 as a blue-violet solid in 87% yield.
1 H NMR(400MHz,CDCl 3 )δ6.80(d,J＝8.0Hz,1H),6.31(d,J＝8.0Hz,1H),4.47(t,J＝4.0Hz,2H),4.36(t,J＝4.0Hz,2H),3.18(t,J＝4.0Hz,2H),3.17(m,2H)； 13 C NMR(101MHz,CDCl 3 )δ143.6,140.8,122.9,121.8,109.2,100.6,65.7,65.0,46.5,46.4；HRMS(ES + )calcd for C 10 H 10 BrNO 2 [M+H] + ,255.9968；Found,255.9970.
(4) Synthesis of Compound 41: 21.8mL of phosphorus oxychloride (234.3 mmol,2 equiv.) are added to 150mL of N, N-dimethylformamide and reacted for 40 minutes on ice before transferring to a solution of 30g of compound 40 (117.1 mmol,1 equiv.) in 1, 2-dichloroethane. The reaction solution was heated to 80℃and reacted for 6 hours. Cool to room temperature and pour into ice water. Suction filtration gave 28.3g of yellow solid product 41 in 85% yield.
1 H NMR(400MHz,CDCl 3 )δ10.16(s,1H),7.10(s,1H),4.48–4.42(t,J＝4.0Hz,2H),4.36–4.30(t,J＝4.0Hz,2H),3.34(m,4H)； 13 C NMR(151MHz,CDCl 3 )δ196.0,142.4,139.4,128.8,123.1,109.8,107.9,64.8,64.1,46.1,46.0；HRMS(ES + )calcd for C 11 H 10 BrNO[M+H] + ,283.9917；Found,283.9925.
(5) Synthesis of Compound 42: 10g of compound 41 (35.2 mmol,1 equiv.), 3.28g of ethylene glycol (52.8 mmol,1.5 equiv.), 5.22g of triethyl orthoformate (35.2 mmol,1 equiv.) and 0.6g of p-toluene sulfonic acid (3.52 mmol,0.1 equiv.) are dissolved in 200mL of toluene. Heating to 100 ℃ for reaction for 12 hours. Cooling to room temperature, neutralizing with saturated sodium bicarbonate, extracting with dichloromethane, drying with anhydrous sodium sulfate, suction filtering, and spin drying. Column chromatography (PE/EA/tea=100/10/1, v/v/v) gave 8.4g of compound 42 as a yellow solid in 73% yield.
1 H NMR(400MHz,CDCl 3 )δ6.68(s,1H),6.03(s,1H),4.48–4.40(m,2H),4.34–4.28(m,2H),4.12–4.04(m,2H),4.03–3.96(m,2H),3.20–3.12(m,4H).
(6) Synthesis of Compound 43: 10g of compound 42 (30.5 mmol,1 equiv.) are dissolved in 200mL dry tetrahydrofuran and 14.6mL n-butyllithium (45.7 mmol,2.5M,1.2 equiv.) are added by syringe at-78deg.C for 20 min. 3.54mL of DMF (45.7 mmol,1.5 equiv.) are added and the reaction is allowed to react for 2 hours at room temperature. The saturated ammonium chloride solution is quenched, extracted by methylene dichloride, dried by anhydrous sodium sulfate, filtered by suction and spun-dried. Column chromatography (PE/EA/tea=100/25/1, v/v/v) gave 7.3g of compound 43 as a yellow solid in 89% yield.
1 H NMR(400MHz,CDCl 3 )δ10.38(s,1H),6.82(s,1H),6.53(s,1H),4.53–4.45(m,2H),4.44–4.39(m,2H),4.14–3.96(m,4H),3.29–3.13(m,4H). 13 CNMR(151MHz,CDCl 3 )δ189.7,149.1,148.4,131.0,115.8,107.2,99.9,65.6,65.2,46.4；46.0；HRMS(ES + )calcd for C 14 H 15 NO 5 [M+Na] + ,300.0842；Found,300.0853.
(7) Synthesis of Compound 44: 10g of compound 43 (36 mmol,2 equiv.) and 2.82g of 1, 4-cyclohexanedione monoethylene glycol ketal (18 mmol,1 equiv.) are dissolved in 20mL of ethanol. 0.5mL of 40% aqueous sodium hydroxide solution was slowly added dropwise to the reaction mixture, and the reaction was stirred for 12 hours. Suction filtration and drying of the filter cake gave 9.9g of compound 44 in 81% yield.
1 H NMR(600MHz,CDCl 3 )δ7.81(s,2H),6.66(s,2H),5.70(s,2H),4.41–4.31(m,8H),4.13–4.04(m,4H),3.97–3.90(m,4H),3.83(s,2H),3.18–3.15(m,4H),3.15–3.12(m,4H),2.72(s,4H). 13 C NMR(151MHz,CDCl 3 )δ186.5,144.1,135.8,134.3,126.6,122.5,115.9,107.3,106.7,65.3,65.2,64.4,46.7；46.4,37.4；HRMS(ES + )calcd for C 36 H 38 N 2 O 11 [M+H] + ,675.2548；Found,675.2553.
(8) Synthesis of Compound 45: 2g of diphenyl ether (11.9 mmol,4 equiv.) are dissolved in 30mL of anhydrous tetrahydrofuran. 4.7mL of n-butyllithium (11.9 mmol,4 equiv.) was slowly added to the syringe under an ice-water bath. After 40 minutes of reaction at 0 ℃, transfer to 100mL of tetrahydrofuran solution in which 2g of compound 44 (2.9 mmol,1 equiv.) was dissolved. The reaction was carried out at 0℃for 2 hours. The saturated ammonium chloride solution is quenched, extracted by methylene dichloride, dried by anhydrous sodium sulfate, filtered by suction and spun-dried. The crude product was dissolved in 50mL of methylene chloride and 2mL of methanesulfonic acid (excess) was added to react for 4 hours. The saturated sodium bicarbonate solution is neutralized, dichloromethane extraction, anhydrous sodium sulfate drying, suction filtration and recrystallization are carried out to obtain 0.88g of compound 45, and the yield is 45%. The mixture was used in the next step without isolation and purification.
(9) Synthesis of Compound 46: 2.6g of 2-bromotoluene (15.2 mmol,10 equiv.) are dissolved in 30mL of anhydrous tetrahydrofuran. 6.1mL of n-butyllithium (15.2 mmol,2.5M,10 equiv.) are added via syringe at-78deg.C and the reaction is stirred for 20 minutes before transferring to a solution of 1g of compound 45 (1.52 mmol,1 equiv.) in 150mL of tetrahydrofuran. The reaction was carried out at 0℃for 2 hours. Adding water to quench the reaction, separating liquid, extracting aqueous phase with dichloromethane, collecting organic phase, drying with anhydrous sodium sulfate, suction filtering, and spin drying. The crude product was dissolved in 50mL of methylene chloride, 2mL of methanesulfonic acid (excess) was added, and the reaction was stirred for 8 hours. Neutralizing with saturated sodium bicarbonate solution, extracting with dichloromethane, drying with anhydrous sodium sulfate, vacuum filtering, and spin drying. Column chromatography (DCM/meoh=20/1, v/v) gave 700mg of compound 46 in 63% yield.
Compound 46 was dissolved to make a 2.5 μm solution in dichloromethane in a 4mL quartz cuvette, the UV2600i was used to collect the UV-vis absorption spectrum of the dye, the Dutta was used to collect the fluorescence emission spectrum of the dye, and the detectors were R928 PMT and INGaAS. The absorption emission spectrum of compound 46 in methylene chloride is shown in fig. 11.
1 H NMR(600MHz,CDCl 3 )δ7.60(t,J＝7.4Hz,1H),7.56–7.47(m,2H),7.45–7.34(m,5H),7.30(d,J＝8.2Hz,2H),7.24(s,2H),6.87(dt,J＝13.9,7.4Hz,2H),6.78–6.63(m,4H),4.33(m,8H),3.73(m,8H),2.24(s,2H)； 13 C NMR(151MHz,CDCl 3 )δ148.9,130.7,129.9,129.8,129.7,128.3,126.0,125.0,123.8,123.8,117.3,117.2,109.9,64.1,63.8,47.3,47.2,31.5,19.9；HRMS(ES + )m/z calcd for C 49 H 37 N 2 O 5 [M] + ,733.2697；Found,733.2701.
Example 16: super-resolution fluorescence microscopic imaging of nano-microspheres
Compound 34 was prepared as a 40 μm, 0.5mL aqueous solution, added to a 30 μl suspension of polystyrene nanobeads at a concentration of 2.5wt% (polystyrene nanobeads suspension purchased from tin-free regex biotechnology ltd.) and shaken for 5 minutes, the solution was diluted in a volume ratio of 1/100, the diluted solution was sucked by a pipette, dropped onto a cover glass, evaporated to dryness, and then glycerin was dropped onto a glass slide. By lambda ex ＝808nm、λ em Structured light evident micro imaging (SIM) and wide field imaging were performed =830 nm.
The result of super-resolution imaging of compound 34 for nanospheres is shown in fig. 12. A represents the contrast of wide-field imaging and structured light obvious micro-imaging (SIM) super-resolution imaging of microspheres with different nano-particle diameters. B represents the imaging result of the denser 300-nanometer polystyrene nanometer microsphere. C and D are enlarged views of the boxed area in B, where C is the effect of SIM super-resolution imaging and D is the effect of conventional wide-field fluorescence microscopy imaging. Experiments show that the compound has the potential of near infrared (> 800 nm) super-resolution fluorescence microscopic imaging.
Example 17: cell level super-resolution fluorescence microscopy imaging
Compound 36 was formulated as 1. Mu.M PBS buffer withPolyoxymethylene-immobilized U2OS cells and non-immobilized living cells were incubated for about 30 min, and after incubation, U2OS cells were subjected to structured light imaging (SIM) and wide field imaging using lambda ex ＝808nm、λ em =830 nm shooting.
Compound 36 was used for SIM super-resolution microscopy imaging and wide field imaging of fixed cell (U2 OS cell line) mitochondria and living endocytosis as shown in fig. 13. The upper right hand corner inset of the right hand figure is a bright field image of living cells. Experiments show that the compound has the potential of near infrared (> 800 nm) cell level super-resolution fluorescence microscopic imaging.
Example 18: in vivo fluorescence imaging of mice
The nanomicelles of compounds 28, 24, 34 and DSPE-mPEG2000 entrapped compound 34 were dissolved in PBS buffer to prepare an injection at a concentration of 50 mM. Injecting the injection into body via tail vein, anesthetizing the mouse with isoflurane gas, placing on near infrared living body fluorescence imaging platform, and administering lambda ex Excitation light of 808nm and long-pass filter photographing of different wavelengths.
In vivo fluorescence imaging of mice injected with compounds 28, 24, 34 and DSPE-mPEG2000 nanomicelle after entrapment of compound 34 is shown in fig. 14. Different long pass filters (LP) and Exposure Times (ET) are used, respectively, and imaging is performed after different injection times (PI). Near infrared two-region in vivo fluorescence imaging is performed on the Liver (lever), spleen (Spleen), duodenum (duodenonum), small intestine (Small intestine), saphenous vein system (Saphenous vasculature), abdominal vascular system (Abdominal vasculature), sternum (Sternum) and Tibia (Tibia) of mice, and the experiment shows that the compound has application potential of near infrared in vivo fluorescence imaging.
Example 19: photo-thermal conversion performance
Compound 17 was prepared as a 5mM acetonitrile mother liquor, and the mother liquor was diluted with water to give aqueous solutions having concentrations of 5. Mu.M, 10. Mu.M, 15. Mu.M, 20. Mu.M, 40. Mu.M, 80. Mu.M. In this system, a power density of 0.2W/cm was used 2 ，0.4W/cm 2 ，0.6W/cm 2 ，0.8W/cm 2 ，1W/cm 2 The 808nm laser irradiates, the distance of the light source is about 10cm, the high-sensitivity temperature probe stretches into the center of the solution, the temperature rising process of the laser irradiates for 10min under different dye concentrations is recorded in real time by using a camera, the temperatures of the solutions in different time points are taken, and the temperature change curve of the solutions along with time is drawn.
The photothermal temperature rise curve of the compound 17 is shown in fig. 15, which shows that the compound of the invention has application potential in near infrared photothermal ablation tumor treatment.
Example 20: the compound of the application is excited to generate singlet oxygen
Singlet oxygen detector DPBF was prepared as a 20. Mu.M acetonitrile solution, and its ultraviolet-visible absorption spectrum was measured. Compound 21 was prepared as a 2 μm acetonitrile solution and its absorption spectrum was tested. A mixed solution of 20. Mu.M of singlet oxygen detector DPBF and 2. Mu.M of compound 21 in acetonitrile was prepared and tested for its ultraviolet-visible absorption spectrum. The mixed solution was subjected to a power density of 1W/cm 2 The ultraviolet-visible absorption spectrum of the mixed solution was tested after irradiation with a laser of 808nm for various durations (0 s,5s,10s,20s,30 s). Passing a 20. Mu.M solution of DPBF in acetonitrile through a 1W/cm reactor 2 After irradiation with a 808nm laser, the ultraviolet absorption spectrum was measured.
The generation of singlet oxygen after irradiation of compound 21 with 808nm laser was detected using singlet oxygen detector DPBF, and the experimental results are shown in fig. 16. In A, the 0s irradiation curve and the 30s irradiation curve overlap, and the acetonitrile solution of DPBF with the concentration of 20 μm passes through 1W/cm 2 After irradiation with the 808nm laser, the absorbance of DPBF did not change, indicating that the absorbance spectrum of the DPBF detector did not change when the irradiation was performed in the presence of only DPBF. B represents the absorption spectrum of 20. Mu.M singlet oxygen detector DPBF and 2. Mu.M compound 21 in acetonitrile solution and the absorption spectrum after mixing them, respectively, indicating that the absorption peak at 400nm wavelength is almost the absorption contribution of DPBF. C represents that a mixed solution of 20 mu M of singlet oxygen detector DPBF and 2 mu M of acetonitrile of compound 21 is subjected to 1W/cm for different durations 2 The absorption spectrum after irradiation of the 808nm laser of (2), wherein the 20s irradiation curve and the 30s irradiation curve overlap. After 0s, 5s, 10s, 20s irradiation, the absorbance at the absorption wavelength of DPBF was sequentially measuredDecreasing. C shows that the absorbance of the singlet oxygen detector DPBF at 400nm is obviously reduced, which indicates that the compound 21 generates singlet oxygen after illumination. The test results show that the compound 21 has application potential in photodynamic therapy of diseases.

Claims (10)

1. A compound of formula a:
wherein:
R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 and R is 8 Each independently selected from: hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- ，-SO 2 X，-SO 2 NH 2 And optionally 1 or 2 are selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 An alkyl group; or R is 1 And R is R 2 、R 2 And R is R 3 Or R 3 And R is R 4 And/or R 5 And R is R 6 、R 6 And R is R 7 Or R 7 And R is R 8 Together with the C to which they are each attached, form a group optionally consisting of 1 to 3 groups selected from halogen, C 1-4 Alkyl and C 1-4 Benzene ring substituted by substituent of alkoxy;
R 9 、R 10 、R 11 、R 12 、R 13 each independently selected from: hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- ，-SO 2 X，-SO 2 NH 2 Optionally 1 to 5 are selected from halogen, C 1-4 Alkoxy and azido substituents substituted C 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 Alkyl, optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene substituted aryl C 1-4 An alkoxy group;
R 14 and R is 15 Each independently selected from H, C optionally substituted with halogen 1-6 Alkyl, C 6-14 Aryl and C 2-4 Alkenyl groups; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a group which is optionally substituted with 1 to 5 groups selected from C 1-6 Alkyl, halogen and oxo substituted 3-8 membered heterocyclyl or 4-15 membered heterobridged heterocyclyl; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a 3-8 membered cycloalkyl spiro 4-15 membered heterobridged ring group or a 3-8 membered heterocyclyl spiro 4-15 membered heterobridged ring group; the heterocyclyl and heterobridged ring radicals have 1 to 5 heteroatoms selected from N, O and S;
R 16 and R is 17 Each independently selected from H, C optionally substituted with halogen 1-4 Alkyl, C 1-4 Alkoxy and C 2-4 Alkenyl groups; or R is 16 C, R to which it is attached 14 、R 14 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 5-7 membered nitrogen-containing heterocycle substituted by substituent of alkyl, and/or R 17 C, R to which it is attached 15 、R 15 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 A 5-7 membered nitrogen-containing heterocycle substituted with a substituent of an alkyl group;
a-is acid ion for removing proton, and the acid is one or more selected from HCl, trifluoroacetic acid, methanesulfonic acid, acetic acid and sulfuric acid;
x is halogen;
y is O or S;
n is an integer selected from 1-500.
2. The compound of claim 1, wherein R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 And R is 8 Each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 And optionally 1 or 2 selected from C 1-4 C substituted with substituents of alkoxy, halogen, azido, amino and mercapto groups 1-4 An alkyl group; preferably, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 And R is 8 Each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 1-4 Alkyl, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 The method comprises the steps of carrying out a first treatment on the surface of the More preferably, R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 And R is 8 Each independently selected from hydrogen, halogen, C 1-4 Alkyl, -SO 3- and-SO 2 X, X is as defined in claim 1.
3. The compound of claim 1, wherein R 9 、R 10 、R 11 、R 12 、R 13 Each independently selected from hydrogen, amino, hydroxy, nitro, cyano, carboxyl, halogen, aryl C 1-4 Alkoxy, C 2-4 Alkenyl, -SO 3- 、-SO 2 X、-SO 2 NH 2 C optionally substituted with 1-5 substituents selected from halogen and azido 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 Alkoxy, halogen and azido substituted C 1-4 Alkyl, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 An alkoxy group; preferably, R 9 、R 10 、R 11 、R 12 、R 13 Each independently selected from hydrogen, -SO 3- 、-SO 2 X、-SO 2 NH 2 Optionally from 1 to 3Halogen and azido substituted C 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 Alkoxy, halogen and azido substituted C 1-4 Alkyl, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 An alkoxy group; more preferably, R 9 、R 10 、R 11 、R 12 、R 13 Each independently selected from: hydrogen, -SO 3- C optionally substituted with 1-3 substituents selected from halogen and azido 1-4 Alkoxy, optionally substituted with 1 or 2 groups selected from C 1-4 Alkoxy and halogen substituents 1-4 Alkyl, and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy, X, Y and n are as defined in claim 1.
4. The compound of claim 1, wherein the oxo-heterocyclic group is 1, 2, or 3 oxo-heterocyclic group; the oxo-heterocyclyl is a 5-7 membered heterocyclyl having 1-3 heteroatoms selected from O, S and N; n is an integer selected from 1-300; preferably, n is an integer selected from 1-200.
5. The compound of claim 1, wherein R 14 And R is 15 Each independently selected from H, C 1-6 Alkyl and C 6-14 An aryl group; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a group which is optionally substituted with 1 to 3 groups selected from C 1-6 Alkyl, halogen and oxo substituted 4-7 membered heterocyclyl or 5-12 membered heterobridged heterocyclyl; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a 4-7 membered heterocyclyl spiro 5-12 membered heterobridged ring group; the heterocyclyl and heterobridged ring radicals have 1 to 3 heteroatoms selected from N, O and S.
6. The compound of claim 1, wherein R 16 And R is 17 Each independently selected from H, C 1-4 Alkyl and C 1-4 An alkoxy group; or R is 16 C, R to which it is attached 14 、R 14 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 5-7 membered nitrogen-containing heterocycle substituted by substituent of alkyl, and/or R 17 C, R to which it is attached 15 、R 15 The N groups linked together form a group which is optionally selected from C by 1 to 4 1-4 Alkyl and halogenated C 1-4 A 5-7 membered nitrogen-containing heterocycle substituted with a substituent of an alkyl group.
7. A compound according to claim 1 wherein,
R 1 、R 2 、R 3 、R 4 、R 5 、R 6 、R 7 and R is 8 Each independently is hydrogen, -SO 3- 、C 1-4 An alkyl group;
R 9 、R 10 、R 12 、R 13 each independently selected from hydrogen, -SO 3- And optionally C 1-4 Alkoxy substituted C 1-4 An alkyl group; r is R 11 Selected from hydrogen, C optionally substituted by 1-3 substituents selected from halogen and azido 1-4 Alkoxy and optionally oxo-heterocyclyl-OC (=o) -C 1-4 alkylene-Y- [ C 1-4 alkylene-O] n -C 1-4 Alkylene-substituted heteroaryl C 1-4 Alkoxy, wherein the oxo-heterocyclic group is dioxo 4-6 membered heterocyclic group, Y is O, and n is an integer selected from 1-100;
R 14 and R is 15 Each independently selected from H, C 1-6 Alkyl and C 6-10 An aryl group; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a group which is optionally substituted with 1 to 3 groups selected from C 1-6 Alkyl, halo and oxo substituted 4-6 membered heterocyclyl or 6-9 membered heterobridged heterocyclyl; or R is 14 And R is 15 Together with the nitrogen atom to which they are attached, form a 4-6 membered heterocyclyl spiro 6-9 membered heterobridged ring group; the heterocyclyl and heterobridged ring radicals have 1-2 heteroatoms selected from N, O and S;
R 16 and R is 17 Each independently of the otherSelected from H and C 1-4 An alkoxy group; or R is 16 C, R to which it is attached 14 、R 14 The N groups bound together form a 5-to 7-membered nitrogen-containing heterocycle, and/or R 17 C, R to which it is attached 15 、R 15 The attached N together form a 5-7 membered nitrogen containing heterocycle; preferably, R 16 And R is 17 Is C 1-4 Alkoxy, R 16 C, R to which it is attached 14 、R 14 The N groups attached together form a 5-7 membered nitrogen containing heterocycle, and R 17 C, R to which it is attached 15 、R 15 The attached N's together form a 5-7 membered nitrogen containing heterocycle.
8. The compound of claim 1, wherein the compound is selected from the group consisting of:
9. a fluorescent dye, dye composition or kit comprising a compound of any one of claims 1-8 and optionally a solvent; preferably, the solvent is selected from one or more of MeOH, etOH, prOH, iPrOH, buOH, water, acetonitrile, acetone, DMF, DMSO, pyridine, dichloromethane, chloroform, dichloroethane, benzene, toluene, para-xylene, chlorobenzene, nitrobenzene, 1, 4-dioxane, THF, ethyl acetate, acOH and buffers; preferably, the buffer is a PBS buffer.
10. Use of a compound according to any one of claims 1-8 for the preparation of an agent for near infrared fluorescence imaging, photothermal therapy and/or photodynamic therapy.
CN202311639934.5A 2023-12-01 2023-12-01 Infrared fluorescent dye and application thereof Pending CN117658973A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202311639934.5A CN117658973A (en) 2023-12-01 2023-12-01 Infrared fluorescent dye and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202311639934.5A CN117658973A (en) 2023-12-01 2023-12-01 Infrared fluorescent dye and application thereof

Publications (1)

Publication Number Publication Date
CN117658973A true CN117658973A (en) 2024-03-08

Family

ID=90065503

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202311639934.5A Pending CN117658973A (en) 2023-12-01 2023-12-01 Infrared fluorescent dye and application thereof

Country Status (1)

Country Link
CN (1) CN117658973A (en)

Similar Documents

Publication Publication Date Title
CN110461327B (en) Diagnostic and therapeutic agents
CN108714222B (en) Preparation and application of novel multifunctional supermolecule vesicle
CN111196819B (en) D-A-D type benzopyrazines compound, preparation method and application thereof
CN109796483B (en) Water-soluble cationic photosensitizer and preparation and application thereof
CN112194787B (en) Preparation method and application of degradable copolymer with near-infrared two-region photoresponse
CN108727256A (en) A kind of photosensitizer and the preparation method and application thereof based on the more pyridiniujms of triphenylamine
JP2023539353A (en) Tumor contrast compound, its preparation method and application to tumor diagnostic imaging
WO2013131235A1 (en) Two-photon fluorescent probe using naphthalene as matrix and preparation method and use thereof
CN111592482B (en) PH reversible activation type photo-thermal/photodynamic/fluorescent integrated probe molecule
CN113717089B (en) Fluorinated Cy7 compound and synthetic method and application thereof
CN113956265A (en) Near-infrared molecular probe based on malondialdehyde response, preparation method and application thereof
CN114105982B (en) Near infrared dye based on naphthalimide, preparation and application thereof
CN108558788A (en) A kind of novel fluorescent dye synthetic method with aggregation-induced emission phenomenon
CN112442019B (en) Seven-methine functional cyanine dye based on click activation and large steric hindrance aggregation quenching resistance, and preparation method and application thereof
CN111303111B (en) Huang Jing near-infrared two-region dye, preparation method and fluorescence imaging application
CN111793371B (en) 3, 5-asymmetrically modified BODIPY near-infrared fluorescent dye and preparation method thereof
CN117658973A (en) Infrared fluorescent dye and application thereof
CN111196821B (en) Compounds, preparation method thereof and application thereof as near-infrared two-region fluorescent probe for detecting methylglyoxal
CN110615755A (en) Near-infrared fluorescent molecule for controlled release of singlet oxygen and preparation method thereof
CN113278036A (en) Phenothiazine-containing iridium complex and preparation method and application thereof
CN111849196B (en) Near-infrared two-region dye and synthetic method thereof
CN114380856B (en) Silorhodamine derivative for detecting brain hydrogen sulfide and preparation method and application thereof
Lioret et al. Valkyrie Probes: A Novel Class of Enzyme‐Activatable Photosensitizers based on Sulfur‐and Seleno‐Rosamines with Pyridinium Unit
RU2725876C1 (en) Derivatives of fluorine-containing chlorins exhibiting anti-tumour activity
CN113004264B (en) Aggregation-induced emission photosensitizer, preparation method and application

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