A diethylene glycol-based amphiphilic molecule, and a preparation method and application thereof

Abstract

The application discloses a diethylene glycol-based amphiphilic molecule and a preparation method and application thereof, relates to the field of molecular recognition, and the structure of the amphiphilic molecule contains a diethylene glycol group as a hydrophilic group, contains a pyrene group with hydrophobicity and luminescence, and further contains a terpyridine group as a metal ion recognition point; the molecule is simple to synthesize, has a high yield, and can be obtained through two-step reactions of nucleophilic addition and esterification. Further ultraviolet titration experiments show that when the molecule is embedded on a two-phase vesicle membrane containing a lipid raft microregion, DEG-2 is combined with Cu 2+ The maximum binding constant of the metal ion indicates that the existence of the lipid raft microregion on the vesicle membrane helps the amphiphilic molecule to be combined with Cu 2+ The application provides the preparation of the amphiphilic molecule and self-assembly of the amphiphilic molecule on a heterogeneous vesicle membrane, and high-efficiency recognition of heavy metal ions in an environment or a plant body can be realized.

Description

Technical Field

[0001] This application relates to the field of molecular recognition technology, specifically to an amphiphilic molecule containing a diethylene glycol group, its preparation method, and its application. Background Technology

[0002] Molecular recognition is a prerequisite and foundation for vital functions such as material transport, signal transduction, and enzyme catalysis in biological systems. Extensive research has revealed that lipid raft microdomains on phospholipid bilayer vesicle membranes play a crucial role in molecular recognition. Lipid raft microdomains are microstructural domains on the cell membrane rich in cholesterol and sphingomyelin, existing in a liquid-ordered phase (Lo), and serving as gateways for information exchange on the cell membrane. Simulating related functions of the cell membrane, an increasing number of research groups have reported interactions between host and guest molecules on homogeneous vesicle membranes, indicating that vesicle membranes can promote host-guest recognition. However, how heterogeneous vesicle membranes, i.e., those containing lipid raft microdomains, affect host-guest interactions, and whether they can further enhance host-guest binding capacity, remains unreported.

[0003] Therefore, this application proposes to design and prepare an amphiphilic molecule based on diethylene glycol, whose structure has the following characteristics: ① Diethylene glycol is used as a hydrophilic group because polyethylene glycol has the characteristics of stability, solubility, biocompatibility and safety. Amphiphilic molecules prepared based on polyethylene glycol can serve as ideal monomers for constructing functional vesicle membranes; ② Pyrene is used as a hydrophobic group because it has strong ultraviolet absorption peaks and fluorescence emission peaks in multiple wavelength bands. When the amphiphilic molecule is embedded in the vesicle membrane, the change in the absorption wavelength of this group can reflect the influence of vesicle membrane inhomogeneity on the self-assembly of the amphiphilic molecule; ③ Tripyridine is used as a metal ion recognition group to further study the effect of lipid raft micro-region vesicle membranes on Cu. 2+ Impact of metal ion recognition. The amphiphilic molecules prepared in this application and the functional vesicle membrane system constructed based on them are expected to achieve efficient detection of metal ions in systems such as plants, soil, environment, and water sources. Summary of the Invention

[0004] This invention provides an amphiphilic molecule containing a diethylene glycol group, its preparation method, and its application. The results of proton, carbon, and high-resolution mass spectrometry show that the prepared substance is consistent with the expected substance and has high purity.

[0005] Technical solution

[0006] An amphiphilic molecule containing a diethylene glycol group, wherein the diethylene glycol-containing amphiphilic molecule is DEG-2, has the following structure:

[0007]

[0008] This application also discloses a method for preparing an amphiphilic molecule containing a diethylene glycol group, the specific steps of which are as follows:

[0009] Step 1, synthesis of compound 2-1: Under nitrogen protection, NaH (25 mg, 1.04 mmol) and DMF (3 mL) were added sequentially to the reaction flask;

[0010] Weigh diethylene glycol (380.0 mg, 3.58 mmol) and DMF (2.0 mL), mix them rapidly, and inject them into a reaction flask. Stir at 200 rpm for 30 min in an ice-water bath.

[0011] Weigh 400.0 mg, 1.35 mmol of bromopyrene and 3.0 mL of DMF, mix them rapidly, and inject them into a reaction flask. Stir the mixture at 200 rpm for 24 h in an ice-water bath.

[0012] The reaction was monitored by thin-layer chromatography (TLC) until the bromopyrene in the mixture was completely reacted, and the criterion for judgment was that the concentration of bromopyrene monitored by TLC was 0.

[0013] The developing solvent used for TLC was ethyl acetate-n-hexane (V / V = 1:3);

[0014] After the reaction was completed, an aqueous solution (15 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (15 mL).

[0015] The extract was dried over anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain the crude product.

[0016] The crude product was separated and purified by silica gel column chromatography. The mobile phases were ethyl acetate-n-hexane (V / V = 1:3, 100 mL), ethyl acetate-n-hexane (V / V = 1:1, 200 mL), and methanol-ethyl acetate (V / V = 1:1, 200 mL). Finally, a light brown oily compound 2-1 (190.0 mg, yield 95%) was obtained.

[0017] The second step, the synthesis of DEG-2: Compound 2-1 (460.0 mg, 1.43 mmol) was added to the reaction flask, and dichloromethane (15 mL) was injected into the reaction flask under nitrogen protection;

[0018] Weigh 4-carboxyphenyl terpyridine (490 mg, 1.38 mmol) and add it to the reaction flask;

[0019] Weigh 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide EDC (400.0 mg, 2.08 mmol) and 4-dimethylaminopyridine DMAP (50.0 mg, 0.41 mmol) and add them to a reaction flask. Stir the mixture at 200 r / min for 24 h in an ice-water bath.

[0020] The reaction was monitored by thin-layer chromatography (TLC) until the 4-carboxyphenyl terpyridine in the mixture was completely reacted. The criterion for judgment was that the concentration of 4-carboxyphenyl terpyridine was 0 as monitored by TLC.

[0021] The developing solvent used for TLC was ethyl acetate-n-hexane (V / V = 1:1);

[0022] After the reaction was completed, methanol and n-hexane solutions were added to the reaction solution, and the crude product was obtained by filtration and rotary evaporation. The amount of methanol and n-hexane solutions added was 10 mL of methanol and 10 mL of n-hexane.

[0023] The crude product was separated and purified using a silica gel column. The mobile phases were ethyl acetate-n-hexane (V / V = 1:2, 100 mL) and ethyl acetate-n-hexane (V / V = 1:1, 150 mL) in sequence. The remaining material in the column was purified using ethyl acetate (100 mL) as the mobile phase.

[0024] Finally, a colorless oily compound, DEG-2 (290.0 mg, yield 63%), was obtained.

[0025] Furthermore, the bromopyrene is 1-(bromomethyl)pyrene.

[0026] Furthermore, the 4-carboxyphenyl terpyridine is 4'-(4-carboxyphenyl)-2,2':6',2"-terpyridine.

[0027] This application also discloses the application of diethylene glycol-based amphiphilic molecules in the preparation of monolayer vesicle membranes. The steps are as follows: DEG-2 (1.0 mM, 5 μL), phospholipid molecules (1.0 mM, 45 μL), and CHCl3 / MeOH (2:1 v / v, 50 μL) are mixed in a 10 mL test tube, and the solvent is evaporated at 37 °C using a rotary evaporator. Then, the phospholipid membrane in the test tube is vacuum dried at 25 °C for 1.5 h, followed by the addition of HEPES buffer solution (20 mM, 1.0 mL, pH = 7.45), followed by shaking at 37 °C for 1 h, and freeze-thawing three times. After vortexing for 10 s, the membrane is incubated in a 37 °C oven for 12 h. Finally, the solution is squeezed through a 0.25 μm filter membrane 5-6 times to obtain a monolayer vesicle membrane based on phospholipid molecules.

[0028] Furthermore, the phospholipid molecules are one or more of the following: DOPC phospholipid molecules in a liquid disordered phase (Ld), DMPC / chol phospholipid molecules in a liquid ordered phase (Lo), and DOPC / Chol / sm phospholipid molecules that have both liquid ordered and disordered phases.

[0029] This application also discloses a method for using diethylene glycol-containing amphiphilic molecules in Cu. 2+ For applications in metal ion recognition, the following steps are first prepared: a DMSO solution of DEG-2 (5 μM, 1 mL), an aqueous solution of a vesicle membrane containing DEG-2 (10 μM, 1 mL), and CuCl2 (5 × 10⁻⁶). -4 An aqueous solution of M) was used as the stock solution for ultraviolet titration.

[0030] A diethylene glycol-containing amphiphilic molecule in Cu 2+ In the application of metal ion recognition, the amphiphilic molecule containing diethylene glycol groups is DEG-2 on a single-layer vesicle membrane.

[0031] Furthermore, the monolayer vesicle membrane is one or more of the following: a DOPC-based monolayer vesicle membrane, a DMPC / chol phospholipid molecule-based monolayer vesicle membrane, and a DOPC / Chol / sm-based monolayer vesicle membrane.

[0032] Beneficial effects:

[0033] 1. Huang Qiang et al. designed and synthesized a novel antipyrine-Schiff base derivative, which can react with Cu in aqueous solution. 2+ The metal ions form a 1:1 complex with a binding constant of 6.5 × 10⁻⁶. 4 M -1 However, the binding constant of the amphiphilic molecule designed in this application on heterogeneous vesicle membranes is 2.83 × 10⁻⁶. 5 mM -1 It is more than four thousand times that of the former. It can be seen that when amphiphilic molecules are embedded into the vesicle membrane, their binding ability with metal ions can be greatly increased (Journal of Luminescence, 2013, 34(9): 1144-1148).

[0034] 2. Pan's research group studied the interaction between sulfocalixarene derivatives (main molecules) and NH4. +The binding behavior of the guest molecule in solution and on homogeneous vesicle membranes was studied. The results showed that when the host molecule was embedded in the vesicle membrane, its binding ability with the guest molecule increased by 5.2 times (Chinese Chem. Lett. 2017, 28(4), 787-792). In contrast, in the application of the diethylene glycol-containing amphiphilic molecule and its preparation method, when the amphiphilic molecule was embedded in a heterogeneous vesicle membrane, its binding constant with metal ions was 16.8 times that in solution. This indicates that the presence of lipid raft microregions on the vesicle membrane helps the amphiphilic molecule to bind with Cu in the best configuration. 2+ Metal ion binding;

[0035] 3. The amphiphilic molecule designed in this application is simple to synthesize and has a high yield. Previous studies have reported more on the recognition effects of different host molecules and ions in solution, such as fluorescent probes containing polypeptide recognition groups selectively recognizing copper ions and cyanide ions (CN 105542755 B), sulfonic acid indole indigo derivatives selectively recognizing copper ions (CN 108912037 B), and fluorescent probes containing p-toluenesulfonyl hydrazine groups recognizing copper ions and mercury ions (CN 109722241 B). Less research has been conducted on the host-guest recognition properties on vesicle membranes. This application fills this gap, and experimental results show that heterogeneous vesicle membranes help improve the binding ability of host and guest molecules. This invention is expected to be applied to the efficient recognition of heavy metals in the environment and plants. Attached Figure Description

[0036] Figure 1 This is a schematic diagram of the preparation route of the amphiphilic molecule DEG-2 in this application;

[0037] Figure 2 To adopt 1 H NMR, 13 The structure of DEG-2 was characterized by C NMR and HRMS, where (a) is the proton spectrum of DEG-2; (b) is the carbon spectrum of DEG-2; and (c) is the high-resolution mass spectrum of DEG-2.

[0038] Figure 3 To determine the concentrations of Cu in DMSO solution 2+ UV absorption spectrum of DEG-2 at different concentrations;

[0039] Figure 4 Different concentrations of Cu on DOPC vesicle membranes 2+ UV absorption spectrum of DEG-2 at different concentrations;

[0040] Figure 5 Different concentrations of Cu in DMPC / Chol vesicle membranes 2+ UV absorption spectrum of DEG-2 at different concentrations;

[0041] Figure 6 Different concentrations of Cu in DOPC / Chol / SM vesicle membranes 2+ UV absorption spectrum of DEG-2 at different concentrations. Detailed Implementation

[0042] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0044] Example 1

[0045] See appendix Figure 1 The preparation method of amphiphilic molecules containing diethylene glycol groups, the specific steps are as follows:

[0046] Step 1, Synthesis of Compound 2-1: Under nitrogen protection, 5 mg of NaH2 and 3 mL of DMF were added sequentially to the reaction flask;

[0047] Weigh 380.0 mg of diethylene glycol and 2.0 mL of DMF, mix them rapidly, and inject them into a reaction flask. Stir the mixture at 200 rpm for 30 min in an ice-water bath.

[0048] Weigh 400.0 mg of bromopyrene and 3.0 mL of DMF, mix them rapidly, and inject them into a reaction flask. Stir the mixture at 200 r / min for 24 h in an ice-water bath.

[0049] The reaction was monitored by thin-layer chromatography (TLC) until the bromopyrene in the mixture was completely reacted. The criterion for judgment was that the concentration of bromopyrene monitored by TLC was 0.

[0050] The developing solvent used for TLC was ethyl acetate-n-hexane (V / V = 1:3);

[0051] After the reaction was completed, 15 mL of aqueous solution was added to the reaction solution, and the mixture was extracted three times with 15 mL of ethyl acetate.

[0052] The extract was dried over anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain the crude product.

[0053] The crude product was separated and purified by silica gel column chromatography, with the mobile phases being ethyl acetate-n-hexane (V / V = 1:3, 100 mL), ethyl acetate-n-hexane (V / V = 1:1, 200 mL), and methanol-ethyl acetate (V / V = 1:1, 200 mL) in sequence, finally yielding a light brown oily compound 2-1 (190.0 mg, yield 95%); the bromopyrene was 1-(bromomethyl)pyrene.

[0054] The second step, the synthesis of DEG-2: Compound 2-1460.0 mg was added to the reaction flask, and dichloromethane (15 mL) was injected into the reaction flask under nitrogen protection;

[0055] Weigh 490 mg of 4-carboxyphenyl terpyridine and add it to the reaction flask; 4-carboxyphenyl terpyridine is 4'-(4-carboxyphenyl)-2,2':6',2"-terpyridine;

[0056] Weigh 400.0 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and 50.0 mg of 4-dimethylaminopyridine (DMAP) and add them to a reaction flask. Stir the mixture at 200 r / min for 24 h in an ice-water bath.

[0057] The reaction was monitored by thin-layer chromatography (TLC) until the 4-carboxyphenyl terpyridine in the mixture was completely reacted. The criterion for judgment was that the concentration of 4-carboxyphenyl terpyridine monitored by TLC was 0.

[0058] The developing solvent used for TLC was ethyl acetate-n-hexane (V / V = 1:1);

[0059] After the reaction was completed, methanol and n-hexane solutions were added to the reaction solution, and the crude product was obtained by filtration and rotary evaporation. The amount of methanol and n-hexane solutions added was 10 mL of methanol and 10 mL of n-hexane.

[0060] The crude product was separated and purified by silica gel column chromatography. The mobile phases were ethyl acetate-n-hexane (V / V = 1:2, 100 mL) and ethyl acetate-n-hexane (V / V = 1:1, 150 mL) in sequence. The remaining material in the column was purified by ethyl acetate (100 mL) as the mobile phase.

[0061] Finally, a colorless oily compound, DEG-2 (290.0 mg, yield 63%), was obtained. The experiment used... 1 HNMR, 13 The structure of DEG-2 was characterized by CNMR and HRMS, and the results are as follows: Figure 2 As shown in the figure. The results indicate that the prepared substance conforms to the expected chemical structure and has high purity.

[0062] Example 2

[0063] The application of diethylene glycol-containing amphiphilic molecules in the preparation of monolayer vesicle membranes involves the following steps: DEG-2 (1.0 mM, 5 μL), phospholipid molecules (1.0 mM, 45 μL), and CHCl3 / MeOH (2:1 v / v, 50 μL) are mixed in a 10 mL test tube, and the solvent is evaporated by rotary evaporation at 37 °C. The phospholipid membrane in the test tube is then vacuum dried at 25 °C for 1.5 h, followed by the addition of HEPES buffer solution (20 mM, 1.0 mL, pH = 7.45). The mixture is then shaken at 37 °C for 1 h, and subjected to three freeze-thaw cycles. After vortexing for 10 s, the mixture is incubated in a 37 °C oven for 12 h. Finally, the solution is squeezed through a 0.25 μm filter membrane 5-6 times to obtain a phospholipid-based monolayer vesicle membrane.

[0064] The phospholipid molecules are one or more of the following: DOPC phospholipid molecules in a liquid disordered phase (Ld), DMPC / chol phospholipid molecules in a liquid ordered phase (Lo), and DOPC / Chol / sm phospholipid molecules that have both liquid ordered and disordered phases.

[0065] DOPC phospholipid molecules are unsaturated phosphate esters with a phase transition temperature of -17℃. Therefore, under normal temperature conditions, DOPC-based monolayer vesicle membranes are in a liquid disordered phase (Ld).

[0066] When DOPC phospholipid molecules are replaced with DMPC\chol phospholipid molecules, a monolayer vesicle membrane based on DMPC\chol phospholipid molecules is obtained. Since DMPC is a saturated phosphate ester, its phase transition temperature is 24℃. When it is mixed with cholesterol (chol), the monolayer vesicle membrane based on DMPC\chol phospholipid molecules is in a liquid ordered phase (Lo).

[0067] DOPC phospholipid molecules are replaced by DOPC\Chol\sm phospholipid molecules. Since the phase transition temperature of DOPC phospholipid molecules is -17℃ and the phase transition temperature of sphingomyelin (sm) is 45℃, when they are mixed with cholesterol, the monolayer vesicle membrane based on DOPC\Chol\sm is a heterogeneous membrane (Ld / Lo) in which liquid disordered phase and liquid ordered phase coexist.

[0068] Example 3

[0069] Amphiphilic molecules containing diethylene glycol groups in Cu 2+ For applications in metal ion recognition, the following steps are first prepared: a DMSO solution of DEG-2 (5 μM, 1 mL), an aqueous solution of a vesicle membrane containing DEG-2 (10 μM, 1 mL), and CuCl2 (5 × 10⁻⁶). -4 An aqueous solution of M) was used as the stock solution for UV titration; 2-10 μL of the stock solution was added dropwise to the DEG-2 solution each time, and then the spectrum was scanned; DEG-2 and Cu 2+The binding constants of metal ions in DMSO solution or DEG-2-containing vesicle membrane solution were calculated using UV titration fitting curve software.

[0070] Example 4

[0071] DEG-2 and Cu in DMSO solution 2+ Studies on metal-ion interactions Figure 3 For different concentrations of Cu 2+ The changes in the UV absorption spectrum of DEG-2 under the present conditions (in DMSO solution, concentration 5 μM). As shown in the figure, DEG-2 has three absorption peaks at wavelengths of 277 nm, 327 nm, and 349 nm. The first absorption peak is attributed to the pyrene group, while the latter two peaks may be attributed to bipyridine. With the presence of Cu... 2+ With increasing concentration, the absorbance at 277 nm gradually decreases, and a slight red shift is observed at the absorption peak position at 277 nm; the absorbance at 327 nm and 349 nm gradually increases, with a slight blue shift; an isoabsorption point is observed at 287 nm; the change in absorbance at 315 nm is correlated with Cu 2+ A graph of ion concentrations was plotted to show the relationship between absorbance and Cu. 2+ The concentration exhibits a first-order functional relationship, with the regression equation y = -0.11*exp(-x / -14.93) + 0.10; combined with the constant Ks = 1.68 × 10⁻⁶. 4 mM -1 R = 0.986 (Table 1).

[0072] Example 5

[0073] DEG-2 and Cu on DOPC vesicle membrane 2+ Studies on metal-ion interactions Figure 4 For different concentrations of Cu 2+ The changes in the UV absorption spectrum of DEG-2 under the presence of Cu (on a DOPC vesicle membrane, concentration 10 μM). As shown in the figure, DEG-2 has an absorption peak at 220 nm; with the presence of Cu... 2+ With increasing concentration, the absorbance at 220 nm gradually increases, and a slight blue shift is observed at the absorption peak position at 220 nm; the absorbance at 277 nm gradually decreases, and a slight red shift is observed at the absorption peak position at 277 nm; the change in absorbance at 215 nm is compared with that at Cu. 2+ A graph of ion concentrations was plotted to show the relationship between absorbance and Cu. 2+ The concentration exhibits a first-order functional relationship, with the regression equation y = -0.01*exp(-x / -3.18) - 0.01; combined with the constant Ks = 9.9 × 10 2 mM -1 R = 0.953 (Table 1).

[0074] Example 6

[0075] DEG-2 and Cu in DMPC / Chol vesicle membranes 2+ Studies on metal-ion interactions Figure 5 For different concentrations of Cu 2+ The changes in the UV absorption spectrum of DEG-2 under the presence of Cu (in DMPC / Chol vesicle membrane, concentration 10 μM). As shown in the figure, DEG-2 has an absorption peak at 216 nm; with the presence of Cu... 2+ With increasing concentration, the absorbance at 216 nm gradually increases; the change in absorbance at 222 nm is compared with that at Cu. 2+ A graph of ion concentrations was plotted to show the relationship between absorbance and Cu. 2+ The concentration exhibits a first-order functional relationship, with the regression equation y = 0.01 * exp(-x / -1.02) - 0.01, combined with the constant Ks = 4.3 × 10⁻⁶. 3 mM -1 R = 0.987 (Table 1).

[0076] Example 7

[0077] DEG-2 and Cu in DOPC / Chol / SM vesicle membranes 2+ Studies on metal-ion interactions Figure 6 For different concentrations of Cu 2+ The changes in the UV absorption spectrum of DEG-2 under the presence of Cu (in DOPC / Chol / SM vesicle membranes, concentration 10 μM). As shown in the figure, DEG-2 exhibits two absorption peaks at wavelengths of 222 nm and 277 nm; with the presence of Cu... 2+ With increasing concentration, the absorbance at 222 nm gradually increases, and a slight blue shift is observed at the absorption peak position at 222 nm; the absorbance at 277 nm gradually decreases, and a slight red shift is observed at the absorption peak position at 277 nm; an isoabsorption point appears at 246 nm; the change in absorbance at 216 nm is compared with that at Cu. 2+ A graph of ion concentrations was plotted to show the relationship between absorbance and Cu. 2+ The concentration exhibits a first-order functional relationship, with the regression equation y = -0.03*exp(-x / 2.51) + 0.03, combined with the constant Ks = 2.83 × 10⁻⁶. 5 mM -1 R = 0.937 (Table 1).

[0078] Table 1 DEG-2 and Cu 2+ Binding constants of metal ions interacting in different media (Ks / mM) -1 )

[0079]

[0080] Example 8

[0081] A two-step synthesis involving substitution and esterification was successfully performed to prepare an amphiphilic molecule (DEG-2) containing a diethylene glycol group. The results of proton NMR, carbon NMR, and high-resolution mass spectrometry showed that the prepared substance was consistent with the expected substance and had high purity. This type of amphiphilic molecule has a bipyridine or terpyridine metal ion recognition group, a hydrophilic diethylene glycol group in the middle, and an alkyl chain or methylpyrene hydrophobic group. This molecule can spontaneously assemble on a phospholipid vesicle membrane.

[0082] By preparing large vesicle membranes containing amphiphilic molecules, the experiment further explored the interaction between the amphiphilic molecules (DEG-2) and metal ions (Cu) using ultraviolet titration. 2+ The interactions between DEG-2 and metal ions in DMSO solution, DOPC vesicle membranes, DMPC / Chol vesicle membranes, and DOPC / Chol / SM vesicle membranes were investigated, and the binding affinity between the amphiphilic molecule and metal ions in different media was calculated using spectral titration fitting software. UV spectroscopy results showed that DEG-2 exhibited three absorption peaks in DMSO solution: 277 nm, 327 nm, and 349 nm. The first peak was attributed to the pyrene group, while the latter two peaks likely belonged to bipyridine. With the addition of metal ions, the absorption peaks at 327 nm and 349 nm increased, and a clear isoabsorption point appeared near 280 nm, indicating the formation of a relatively stable complex between the host and guest molecules. When amphiphilic molecules are embedded in the vesicle membrane, the absorption peak of the amphiphilic molecules becomes a single point, mainly at 216 nm. With the addition of metal ions, the absorption peak at this point shows an increasing trend, and an isoabsorption point also appears at 229 nm. The binding constants of amphiphilic molecules and metal ions in vesicles of different states are calculated and shown in Table 1. The table shows that when amphiphilic molecules are embedded in heterogeneous vesicles, i.e., in the vesicle membrane of lipid raft microregions, their binding with Cu... 2+ The highest binding capacity indicates that the presence of lipid raft microregions on the vesicle membrane helps to change the configuration and distribution of amphiphilic molecules with the drop-in of metal ions, thereby achieving the optimal binding mode.

[0083] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. An amphiphilic molecule containing a diethylene glycol group, characterized in that, The amphiphilic molecule containing diethylene glycol groups is DEG-2, which has the following structure: .

2. A method for preparing the diethylene glycol-containing amphiphilic molecule as described in claim 1, characterized in that, The specific steps are as follows: Step 1, Synthesis of Compound 2-1: Under nitrogen protection, 1.04 mmol of NaH 25 mg and 3 mL of DMF were added sequentially to the reaction flask; Weigh 380.0 mg of diethylene glycol (3.58 mmol) and 2 mL of DMF, mix them rapidly, and inject them into a reaction flask. Stir at 200 rpm for 30 min in an ice-water bath. Weigh 1.35 mmol of 400.0 mg of 1-(bromomethyl)pyrene and 3.0 mL of DMF, mix them rapidly, and inject them into a reaction flask. Stir the mixture at 200 r / min for 24 h in an ice-water bath. The reaction was monitored by thin-layer chromatography (TLC) until the 1-(bromomethyl)pyrene in the mixture was completely reacted, and the criterion for judgment was that the concentration of 1-(bromomethyl)pyrene monitored by TLC was 0. The developing solvent used in TLC was ethyl acetate-n-hexane, wherein the volume ratio of ethyl acetate to n-hexane was 1:

3. After the reaction was completed, 15 mL of aqueous solution was added to the reaction solution, and the mixture was extracted three times with 15 mL of ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain the crude product. The crude product was separated and purified by silica gel column chromatography. The mobile phases were 100 mL of ethyl acetate-n-hexane (1:3 v / v), 200 mL of ethyl acetate-n-hexane (1:1 v / v), and 200 mL of methanol-ethyl acetate (1:3 v / v). Finally, 190.0 mg of light brown oily compound 2-1 was obtained, with a yield of 95%. The second step, the synthesis of DEG-2: 1.43 mmol of compound 2-1 460.0 mg was added to the reaction flask, and 15 mL of dichloromethane was injected into the reaction flask under nitrogen protection; Weigh 1.38 mmol of 490 mg of 4'-(4-carboxyphenyl)-2,2':6',2"-terpyridine and add it to the reaction flask; Weigh 2.08 mmol of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide EDC 400.0 mg and 0.41 mmol of 4-dimethylaminopyridine DMAP 50.0 mg and add them to a reaction flask. Stir the mixture at 200 r / min for 24 h in an ice-water bath. The reaction was monitored by thin-layer chromatography (TLC) until the 4'-(4-carboxyphenyl)-2,2':6',2"-terpyridine in the mixture was completely reacted. The criterion for judgment was that the concentration of 4'-(4-carboxyphenyl)-2,2':6',2"-terpyridine was 0 as monitored by TLC. The developing solvent used in TLC was ethyl acetate-n-hexane, with a volume ratio of ethyl acetate to n-hexane of 1:

1. After the reaction was completed, methanol and n-hexane solutions were added to the reaction solution, and the crude product was obtained by filtration and rotary evaporation. The amount of methanol and n-hexane solutions added was 10 mL of methanol and 10 mL of n-hexane. The crude product was separated and purified using a silica gel column. The mobile phase consisted of 100 mL of ethyl acetate-n-hexane with a volume ratio of 1:1 and 150 mL of ethyl acetate-n-hexane with a volume ratio of 1:

1. The remaining material in the column was purified using 100 mL of ethyl acetate as the mobile phase. Finally, a colorless oily compound, namely 290.0 mg of DEG-2, was obtained, with a yield of 63%.

3. The application of the diethylene glycol-containing amphiphilic molecule as described in claim 1 in the preparation of a monolayer vesicle membrane, characterized in that, The steps are as follows: 5 μL of 1.0 mM DEG-2, 45 μL of 1.0 mM phospholipid molecules, and 50 μL of CHCl3 / MeOH are mixed in a 10 mL test tube, with a CHCl3:MeOH volume ratio of 2:

1. The solvent is evaporated by rotary evaporation at 37 °C. The phospholipid membrane in the test tube is then vacuum dried at 25 °C for 1.5 h. 1.0 mL of 20 mM HEPES buffer solution (pH 7.45) is then added. The mixture is shaken at 37 °C for 1 h, and subjected to three freeze-thaw cycles. After vortexing for 10 s, the mixture is incubated in a 37 °C oven for 12 h. Finally, the solution is squeezed through a 0.25 μm filter membrane 5-6 times to obtain a monolayer vesicle membrane based on phospholipid molecules.

4. The application of the diethylene glycol-containing amphiphilic molecule according to claim 3 in the preparation of monolayer vesicle membranes, characterized in that: The phospholipid molecules are one or more of the following: DOPC phospholipid molecules in the liquid disordered phase Ld, DMPC / chol phospholipid molecules in the liquid ordered phase Lo, and DOPC / Chol / sm phospholipid molecules that have both liquid ordered and disordered phases.

5. A Cu containing a diethylene glycol-based amphiphilic molecule as described in claim 1 for non-disease diagnostic and therapeutic purposes. 2+ Its application in metal ion recognition is characterized by: First, prepare 1 mL of 5 μM DEG-2 DMSO solution, 1 mL of 10 μM DEG-2-containing vesicle membrane aqueous solution, and 5 × 10⁻⁶ solutions. −4 An aqueous solution of CuCl2 in M ​​was used as the stock solution for ultraviolet titration.

6. A Cu containing a diethylene glycol-based amphiphilic molecule as described in claim 1 for non-disease diagnostic and therapeutic purposes. 2+ Its application in metal ion recognition is characterized by: The diethylene glycol-containing amphiphilic molecule is DEG-2 on a single-layer vesicle membrane.

7. Cu containing a diethylene glycol-based amphiphilic molecule as described in claim 6 for non-disease diagnostic and therapeutic purposes. 2+ Its application in metal ion recognition is characterized by: The monolayer vesicle membrane is one or more of the following: DOPC-based monolayer vesicle membrane, DMPC / chol phospholipid molecule-based monolayer vesicle membrane, and DOPC / Chol / sm-based monolayer vesicle membrane.

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