Cell membrane bionic conjugated electrolyte molecule taking porphyrin unit as core as well as preparation method and application of cell membrane bionic conjugated electrolyte molecule

Abstract

According to the cell membrane bionic conjugated electrolyte molecule taking the porphyrin unit as the core as well as the preparation method and the application of the cell membrane bionic conjugated electrolyte molecule taking the porphyrin unit as the core, the optical performance, the biocompatibility and the stability of a probe are remarkably optimized by introducing the porphyrin unit and metal coordination; and a more accurate and reliable tool is provided for cell membrane related research and disease diagnosis. The innovative design not only improves the spectral characteristics and the imaging effect, but also promotes the application of the fluorescence imaging technology in the medical field, and has wide prospects.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a cell membrane biomimetic conjugated electrolyte molecule with porphyrin unit as the core, its preparation method, and its application. Background Technology

[0002] Disease diagnosis plays a crucial role in the early detection and precision treatment of diseases. With the continuous advancement of medical technology, efficient diagnostic methods can not only significantly improve cure rates but also effectively reduce the medical resources and costs required during treatment. Among numerous diagnostic techniques, fluorescence imaging has become a key tool in medical diagnosis due to its high sensitivity, high resolution, and real-time dynamic monitoring capabilities. By precisely labeling specific biomarkers with fluorescent probes, fluorescence imaging can visualize lesions, intuitively displaying the location, extent, and molecular-level pathological information of the lesion area, greatly enhancing the accuracy of diagnosing complex diseases such as tumors, infections, and neurodegenerative diseases. Furthermore, fluorescence imaging offers the advantages of being non-invasive, rapid, and providing dynamic observation, providing clinicians with timely and accurate diagnostic data, supporting the development of personalized treatment plans, improving treatment outcomes, and optimizing patient management. With continuous technological advancements, fluorescence imaging not only plays a vital role in basic medical research but also demonstrates broad application potential in clinical diagnosis, providing strong technical support for precision medicine and personalized treatment.

[0003] To address these needs, patent CN116445157A reports a novel biomimetic fluorescent probe for cell membranes. This probe is designed to closely mimic the phospholipid bilayer, employing a composite structure consisting of a hydrophobic rigid conjugated backbone and a hydrophilic flexible alkyl side chain. However, while this probe demonstrates reliability in cell membrane labeling studies, its structural design still requires optimization. First, the conjugated backbone of the probe reported in patent CN116445157A is a nonlinear structure. When the conjugated units embedded in the cell membrane absorb light energy, they rotate or vibrate, thereby disrupting the structural stability of the cell membrane. Second, existing biomimetic fluorescent probes exhibit broad absorption and emission spectra with wide full width at half maximum (FWHM), posing challenges for fluorescence imaging applications. Broad spectra easily lead to signal crosstalk, reducing the signal-to-noise ratio and image resolution, particularly affecting multicolor imaging and high-precision experiments. Simultaneously, increased background noise and reduced excitation efficiency make it difficult to meet the requirements for high specificity, high resolution, or multichannel detection. These issues limit the application of current membrane-inspired fluorescent probes in fields such as multicolor cell membrane dynamic imaging, in vivo tumor microenvironment imaging, and exosome detection, and further optimization is urgently needed. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the present invention aims to provide a cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit as its core, along with its preparation method and applications. The cell membrane biomimetic fluorescent probe proposed in this invention significantly optimizes the probe's optical properties, biocompatibility, and stability by introducing a porphyrin unit and metal coordination, providing a more precise and reliable tool for cell membrane-related research and disease diagnosis. The cell membrane biomimetic conjugated electrolyte molecule prepared according to this invention can be used as a fluorescent probe to label specific biological membranes, highlighting the signals of target cells or microorganisms for easier detection. Furthermore, porphyrin, as a narrow bandgap donor unit, significantly reduces the molecular bandgap, thereby improving imaging accuracy. This innovative design not only improves spectral characteristics and imaging effects but also promotes the application of fluorescence imaging technology in the medical field, showing broad prospects.

[0005] The objective of this invention is achieved through the following technical solutions.

[0006] This invention discloses a cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit as its core, wherein the cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit as its core has the following structure:

[0007]

[0008] in,

[0009] The Ar is selected from electron-donating groups having an aromatic ring structure;

[0010] The —R1 + Selected from one or more alkyl derivatives or aryl derivatives containing quaternary ammonium salts or sulfonates as terminal groups;

[0011] The M is absent, or is selected from metal atoms;

[0012] X - Selected from —R1 + The oppositely charged ion of the group can react with -R1 + Groups form ion pairs;

[0013] All carbon atoms on the alkyl derivative are not substituted, or one or more carbon atoms are substituted by one or more oxygen atoms, amino, sulfone, carbonyl, aryl, alkenyl, alkynyl, ester, cyano, or nitro groups.

[0014] or

[0015] All hydrogen atoms on the alkyl derivative are not substituted, or one or more hydrogen atoms are substituted by one or more of halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, or alkyne groups.

[0016] The aryl derivative is selected from groups containing one or more benzene ring structures, wherein all hydrogen atoms on the benzene ring are not substituted;

[0017] or

[0018] One or more hydrogen atoms at any position on one or more benzene rings are substituted by one or more of a halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, alkyneic, carboxyl, ester, cyano, or nitro group.

[0019] Furthermore, the aforementioned —R1 + Selected from one or more alkyl groups containing quaternary ammonium salts or sulfonates as terminal groups;

[0020] The number of repeating units of the alkyl group is 1-10.

[0021] Furthermore, the Ar is selected from one or more aromatic monocyclic rings, or aromatic fused rings, and combinations of the aforementioned alternatives.

[0022] Furthermore, Ar is selected from one or more structures connected by the following structures:

[0023]

[0024] in,

[0025] X1 is selected independently from S, O, and Se;

[0026] A is selected from carbon or silicon atoms;

[0027] R2 is selected from aryl derivatives, alkyl derivatives, or hydrogen atoms;

[0028] All carbon atoms on the alkyl derivative are not substituted, or one or more carbon atoms are substituted by one or more oxygen atoms, nitrogen atoms, amino groups, sulfone groups, carbonyl groups, aryl groups, alkenyl groups, alkynyl groups, ester groups, cyano groups, and nitro groups.

[0029] or

[0030] All hydrogen atoms on the alkyl derivative are not substituted, or one or more hydrogen atoms are substituted by one or more of halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, or alkyne groups.

[0031] The aryl derivative is selected from groups containing one or more benzene ring structures, wherein all hydrogen atoms on the benzene ring are not substituted;

[0032] or

[0033] One or more hydrogen atoms at any position on one or more benzene rings are substituted by one or more of a halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, alkyneic, carboxyl, ester, cyano, or nitro group.

[0034] Furthermore, the X - It is selected from one or more of the following: boron-containing organic anions, halogen anions, sulfur-containing organic anions, and acid radical anions.

[0035] Furthermore, M is absent or selected from transition metal atoms.

[0036] Furthermore, M is selected from one or more of zinc, cobalt, nickel, and copper.

[0037] More preferably, the cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core is selected from one of the following structures:

[0038]

[0039] The present invention also provides a method for preparing the cell membrane biomimetic conjugated electrolyte molecule with porphyrin unit as core, comprising the following steps:

[0040] S1. Under the protection of an inert gas, the metal porphyrin containing Ar units and compound 5 are coupled together in the presence of a catalyst to obtain compound 6.

[0041] S2. Compound 6 is treated to introduce ionic groups to obtain either cell membrane biomimetic conjugated electrolyte molecule 1 with porphyrin unit as the core or cell membrane biomimetic conjugated electrolyte molecule 2 with porphyrin unit as the core.

[0042] Compound 5 is selected from the following structures:

[0043]

[0044] The -R3 is selected from one or more alkyl derivatives and aryl derivatives;

[0045] X2 is selected from halogens;

[0046] All carbon atoms on the alkyl derivative are not substituted, or one or more carbon atoms are substituted by one or more oxygen atoms, amino, sulfone, carbonyl, aryl, alkenyl, alkynyl, ester, cyano, or nitro groups.

[0047] or

[0048] All hydrogen atoms on the alkyl derivative are not substituted, or one or more hydrogen atoms are substituted by one or more of halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, or alkyne groups.

[0049] The aryl derivative is selected from groups containing one or more benzene ring structures, wherein all hydrogen atoms on the benzene ring are not substituted;

[0050] or

[0051] One or more hydrogen atoms at any position on one or more benzene rings are substituted by one or more of a halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, alkyneic, carboxyl, ester, cyano, or nitro group.

[0052] Furthermore, in step S1, the temperature of the coupling reaction is 70-80°C.

[0053] Further, in step S2, the ionic group includes one or more of the following: quaternary ammonium salt group, phosphate group, sulfonate group, acetate group, quaternary phosphate group, phosphate ester group, and amino group.

[0054] Furthermore, in step S2, the synthetic route for the cell membrane biomimetic conjugated electrolyte molecule 1 with a porphyrin unit as its core is as follows:

[0055]

[0056] Synthesis of S1-1 and Compound 3:

[0057] Under nitrogen protection, compounds 1 (10-15 mmol) and 2 (10-15 mmol) were dissolved in anhydrous dichloromethane (1400-1800 mL), and trifluoroacetic acid (TFA, 0.5-1.0 mol%) was added as a catalyst. The mixture was stirred at room temperature (20-25 °C) for 4-8 h. Maintaining an inert atmosphere, DDQ (1.2-1.5 equivalents) was added, and stirring continued for 30-60 min. The mixture was concentrated under reduced pressure to 25-40% of its original volume (400-640 mL), and a methanol solution of zinc acetate (Zn(OAc)₂, 5.0-6.0 equivalents) (40-60 mL) was added. The mixture was stirred at room temperature for 10-14 h. The reaction solution was washed with water, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by petroleum ether / dichloromethane (v / v ratio 1:3 to 1:5) column chromatography to obtain compound 3.

[0058] Synthesis of S1-2 and Compound 5:

[0059] Under an inert atmosphere, compound 4 (7-9 mmol), p-bromobenzene (1.4-1.6 equivalents), PdCl2(PPh3)2 (8-12 mol%), and potassium carbonate (K2CO3, 1.8-2.2 equivalents) were added to a tetrahydrofuran / water mixed solvent (volume ratio 1:2 to 1:4, total volume 100-150 mL) and reacted at 70-80 °C for 12-16 h. The reaction solution was purified by column chromatography using petroleum ether / ethyl acetate (volume ratio 1:30 to 1:60) as eluent to give compound 5.

[0060] Synthesis of S1-3 and Compound 6:

[0061] Under a nitrogen atmosphere, compound 3 (350-420 μmol), compound 5 (3.5-4.5 equivalents), tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4, 8-12 mol%), and cesium carbonate (Cs2CO3, 2.8-3.2 equivalents) were added to a tetrahydrofuran / water mixed solvent (volume ratio 1:3 to 1:6, total volume 0.8-1.2 mL) and reacted at 70-80 °C for 40-56 h. The reaction solution was purified by column chromatography using petroleum ether / dichloromethane (volume ratio 4:1 to 6:1) as eluent to give compound 6.

[0062] S1-4. Synthesis of cell membrane biomimetic conjugated electrolyte molecule 1 with porphyrin unit as core:

[0063] Compound 6 (0.10-0.12 mmol) was dissolved in chloroform (25-35 mL), and trimethylamine tetrahydrofuran solution (1.8-2.6 mL, 2.0 M) was added. The mixture was stirred at 55 ± 5 °C for 14-18 h. After the reaction, the solid precipitate at the bottom of the flask was collected, the supernatant was discarded, and the precipitate was washed with chloroform 3-5 times. The solid was dissolved in methanol (12-18 mL), and trimethylamine methanol solution (0.9-1.3 mL, 4.0-4.5 M) was added. The mixture was stirred at 55 ± 5 °C for 14-18 h. The reaction solution was concentrated and dried under vacuum to give compound 7, with a yield of 95-99%.

[0064] Furthermore, in step S2, the synthetic route for the cell membrane biomimetic conjugated electrolyte molecule 2 with a porphyrin unit as its core is as follows:

[0065]

[0066] S2-1. Compound 6 is mixed with concentrated hydrochloric acid in a solvent, and the reaction yields compound 8. The mass ratio of reactants is compound 6: concentrated hydrochloric acid: solvent = 1:(1-20):(50-500). The solvent is selected from dichloromethane, chloroform, tetrahydrofuran, or 1,4-dioxane. The reaction is carried out at 0-40℃ for 10-600s. After the reaction is terminated, 1-5% by mass of compound 6 is added as an organic base quencher (preferably pyridine or triethylamine). The reaction solution is filtered through a silica gel short column, and the concentrated filtrate is purified by recrystallization with methanol or ethanol to obtain compound 8.

[0067] S2-2. Ionize compound 8 to obtain the cell membrane biomimetic conjugated electrolyte molecule 2 with porphyrin unit as the core. Dissolve compound 8 in chloroform or dichloromethane (5-20 mL / g of compound 8), add 1.0-2.5 equivalents of trimethylamine solution (tetrahydrofuran or methanol, 1-4 M concentration), and react at 40-70℃ for 8-24 h. After the reaction is complete, collect the precipitate, wash it with chloroform 3-5 times, dissolve the precipitate in methanol, add 1.0-1.5 equivalents of trimethylamine (2-5 M methanol concentration), and react a second time at 40-60℃ for 8-16 h. Finally, concentrate and dry to obtain compound 9, with a yield of 95-99%.

[0068] The present invention also provides the application of the cell membrane biomimetic conjugated electrolyte molecule with porphyrin unit as core in fluorescent probes, biosynthesis, and catalysis.

[0069] The fluorescent probe comprises liposomes and a cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core;

[0070] The molar ratio of the liposomes to the cell membrane biomimetic conjugated electrolyte molecules with porphyrin units as the core is 1000:5-20.

[0071] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0072] This invention significantly optimizes the molecular design of fluorescent probes by introducing porphyrin units with macrocyclic conjugated structures, overcoming challenges related to phototoxicity and spectral performance. The introduction of porphyrin units ensures linearity while the macrocyclic structure enhances probe rigidity and significantly suppresses free rotation under illumination through steric hindrance. This effect maintains high spatial stability after the probe embeds into the cell membrane, reducing photoperiod disturbances. Simultaneously, as a narrow bandgap donor, porphyrin narrows the full width at half maximum (FWHM) of the probe's absorption spectrum, resulting in more precise absorption and avoiding spectral overlap and crosstalk. More importantly, this invention innovatively utilizes the coordination of the nitrogen atom in the porphyrin ring with metal ions to establish a novel spectral modulation method. This method simplifies the synthesis process, effectively eliminates crosstalk between different target fluorescence signals, and significantly improves the selectivity and accuracy of multi-target detection, thereby significantly expanding the application potential of probes in multiplex fluorescence imaging and multichannel detection.

[0073] Specifically:

[0074] First, the macrocyclic structure of the porphyrin unit enhances the rigidity of the probe and effectively increases the steric hindrance between it and phospholipid molecules, significantly restricting the free rotation of molecules under illumination. This steric hindrance effect enables the probe to maintain high spatial stability after embedding in the cell membrane, thereby reducing the disturbance of the cell membrane caused by molecular motion during photoexcitation. Second, the introduction of the porphyrin unit reduces photoinduced cell membrane damage and improves the safety of the probe in in vivo imaging and long-term dynamic monitoring. Furthermore, the unique macrocyclic structure of porphyrin endows the probe with excellent photophysical properties, exhibiting high fluorescence quantum yield and good photostability, further enhancing imaging performance and resulting in a higher signal-to-noise ratio and better resolution during imaging. Moreover, as a narrow bandgap donor unit, porphyrin effectively reduces the full width at half maximum (FWHM) of the absorption spectrum, making the probe's light absorption behavior more precise and avoiding spectral overlap and signal crosstalk. On the other hand, this invention also pioneers a new method of spectral modulation by coordinating the nitrogen atom in the porphyrin ring with metal ions. This method not only simplifies the probe synthesis process but also effectively avoids crosstalk between fluorescence signals from different targets, improving the selectivity and accuracy of multi-target detection. Through this strategy, the optical performance of the probe is significantly enhanced, expanding its application potential in multiplex fluorescence imaging and multichannel detection. Attached Figure Description

[0075] Figure 1 The photon nuclear magnetic resonance (NMR) spectrum of the product of Example 1 (a cell membrane biomimetic conjugated electrolyte molecule 1 with a porphyrin unit as its core) is shown below.

[0076] Figure 2 The photon nuclear magnetic resonance (NMR) spectrum of the product of Example 2 (a cell membrane biomimetic conjugated electrolyte molecule 2 with a porphyrin unit as its core) is shown below.

[0077] Figure 3 The UV-Vis absorption spectra of the products of Examples 1, 2, 3, and 4 in dimethyl sulfoxide solution;

[0078] Figure 4 The fluorescence spectra of the products of Examples 1, 2, 3, and 4 in dimethyl sulfoxide solution are shown.

[0079] Figure 5 The UV-Vis absorption spectrum of Comparative Example 1 in dimethyl sulfoxide solution;

[0080] Figure 6 The fluorescence spectrum of Comparative Example 1 in dimethyl sulfoxide solution;

[0081] Figure 7 These are confocal micrographs of cells in microorganisms from Examples 1 and 2; where a represents dark field, b represents bright field, and c represents superimposed field.

[0082] Figure 8 Calcein leakage experiment using a liposome model under white light irradiation;

[0083] Figure 9 This study investigates the phototoxicity of microorganisms by incorporating biomimetic conjugated electrolyte molecules under white light irradiation. Detailed Implementation

[0084] To more clearly illustrate the technical solution of the present invention, the following embodiments are provided. Unless otherwise stated, the raw materials, reactions, and post-processing methods appearing in the embodiments are all commercially available raw materials and technical methods well known to those skilled in the art.

[0085] PE, petroleum ether.

[0086] DCM, dichloromethane.

[0087] EA, ethyl acetate.

[0088] Example 1

[0089] The synthetic route for the cell membrane biomimetic conjugated electrolyte molecule 1 with a porphyrin unit core is as follows:

[0090]

[0091] Synthesis of S1-1 and Compound 3:

[0092] CH2Cl2 (1600 mL) was added to a 2000 mL double-necked round-bottom flask. After bubbling under nitrogen for 30 min, compound 1 (2.04 g, 14 mmol) and compound 2 (3.25 g, 14 mmol) were dissolved. TFA (trifluoroacetic acid) (705 μL) was added to the reaction mixture as a catalyst, and the mixture was stirred at room temperature for 6 h. DDQ (2,3-dichloro-5,6-dicyano-1,4-benzoquinone) (4.25 g, 18.75 mmol) was added under nitrogen, and the reaction was continued at room temperature for 30 min. The reaction mixture was concentrated to 500 mL by vacuum distillation. Zn(OAc)2 (5.6 g, 70 mmol) was dissolved in 50 mL of methanol solution and added to the reaction system. The mixture was stirred at room temperature for 12 h. The reaction mixture was washed with deionized water, the organic layer was separated and dried with anhydrous Na2SO4, the solvent was removed under reduced pressure, and the mixture was subjected to column chromatography with PE:DCM = 1:4 (v / v) as the eluent to give a purple solid (1.99 g, 41%).

[0093] 1 H NMR (400MHz, CDCl3-pyridine-d5, rt): δH=10.22(s,2H),9.37(d,J=4.4Hz,4H),9 .06(d,J=4.4Hz,4H),8.28(d,J=7.8Hz,4H),8.21(d,J=7.8Hz,4H),1.53(s,24H).

[0094] Synthesis of S1-2 and Compound 5:

[0095] Compound 4 (6 g, 8.09 mmol), p-bromobenzene (2.86 g, 12.14 mmol), PdCl2(PPh3)2 (0.57 g, 0.81 mmol), K2CO3 (2.23 g, 16.19 mmol), and THF:H2O = 1:3 (v / v) (120 mL) were added to a reaction flask. The atmosphere was purged three times, each time for 5 min. The mixture was heated to 75 °C and reacted overnight. Column chromatography was performed using PE:EA = 1:50 (v / v) as the eluent to give a pale yellow oily solid (5 g, 83%).

[0096] 1 H NMR (400MHz, CDCl3): δH=7.53(d,J=8.2Hz,2H),7.39(d,J=8.1Hz,2H),6.70(s,2H ),4.09-3.91(m,6H),3.49-3.77(m,6H),1.93-1.73(m,12H),1.58-1.46(m,12H).

[0097] Synthesis of S1-3 and Compound 6:

[0098] Compound 3 (300 mg, 385.68 μmol), compound 5 (1.2 g, 1.54 mmol), Pd(PPh3)4 (28.22 mg, 38.57 μmol), Cs2CO3 (159.91 mg, 1.16 mmol), and THF:H2O = 1:5 (v / v) (1 mL) were added to a nitrogen-filled reaction flask and refluxed at 75 °C for 48 h. Column chromatography was performed using PE:DCM = 5:3 (v / v) as the eluent to give a purple solid (480 mg, 40%).

[0099] 1 H NMR (400MHz, CDCl3): δH=10.36(d,2H),9.43(d,J=4.6Hz,4H),9.19(d,J=4.6Hz,4H),8.37(d,J=7.6Hz,4H),8.08(d,J=7.7Hz,4H),8.01 (d,J=7.8Hz,4H),7.80(d,J=7.8Hz,4H),6.93(s,4H),4.18-4.03(m,12H),3.51-3.42(m,12H),2.00-1.79(m,24H),1.65-1.54(m,24H).

[0100] Synthesis of S2 and compound 7 (cell membrane biomimetic conjugated electrolyte molecule 1 with porphyrin unit as core):

[0101] Compound 6 (180 mg, 0.1072 mmol) and 30 mL of chloroform were added to a reaction flask. 2.2 mL of trimethylamine (2 M) THF solution was added to the reaction mixture, and the mixture was stirred at 55 °C for 16 h. After the reaction was complete, the crude product precipitated and adhered to the bottom of the flask. The supernatant was discarded, and the solid precipitate was washed five times with chloroform. The solid precipitate was dissolved in 15 mL of methanol. 1.1 mL of trimethylamine (4.2 M) methanol solution was added to the solution, and the mixture was stirred at 55 °C for 16 h. The solution was then evaporated to dryness under vacuum. The final product was a purple solid (218 mg, 97%).

[0102] The proton NMR spectrum of compound 7 is shown below. Figure 1 As shown; the UV-Vis absorption spectrum of compound 7 in dimethyl sulfoxide solution is as follows. Figure 3 As shown: its maximum absorption wavelength is 421 nm, the Q-band absorption peaks are at 550 and 589 nm, its full width at half maximum (FWHM) is 15 nm, and its optical band gap is 2.05 eV. The fluorescence spectrum of compound 7 in dimethyl sulfoxide solution is shown below. Figure 4 As shown: λ(nm): 637,697;

[0103] 1H NMR (400MHz, DMSO): 10.37 (d, J = 5.6 Hz, 2H), 9.53 (d, J = 4.4 Hz, 4H), 9.05 (d, J = 4.3 Hz,4H),8.36(d,J=7.7Hz,4H),8.21(d,J=7.8Hz,4H),8.14(d,J=8.2Hz,4H),7.97 (d,J=7.9Hz,4H),7.09(s,4H).4.18-4.16(m,8H),3.99-3.96(m,4H),3.99-3.96( m,12H),3.08(s,54H),1.85-1.72(m,24H),1.61-1.52(m,12H),1.41-1.3(m,12H).

[0104] Example 2

[0105] The synthetic route for membrane-inspired conjugated electrolyte molecule 2 with a porphyrin unit core is as follows:

[0106]

[0107] Synthesis of S2-1 and Compound 8:

[0108] Compound 6 (200 mg, 50 μmol) was dissolved in dichloromethane (100 mL). Concentrated hydrochloric acid (3 mL) was added, and the resulting green solution was stirred vigorously for 30 s. The reaction mixture was quenched with pure pyridine and passed through a silica gel stopper washed with dichloromethane. All the purple fraction was collected, and after evaporating the solvent, it was recrystallized from methanol. Yield: 198 mg (45 μmol, 90%).

[0109] 1 H NMR (400MHz, CDCl3): δH=10.36(d,2H),9.43(d,J=4.6Hz,4H),9.19(d,J=4.6Hz,4H),8.37(d,J=7.6Hz,4H),8.08(d,J=7.7Hz,4H),8.01(d,J=7 .8Hz,4H),7.80(d,J=7.8Hz,4H),6.93(s,4H),4.18-4.03(m,12H),3.5 1-3.42(m,12H),2.00-1.79(m,24H),1.65-1.54(m,24H),-3.10(s,2H).

[0110] Synthesis of S2-2 and compound 9 (a cell membrane biomimetic conjugated electrolyte molecule 2 with a porphyrin unit core):

[0111] Compound 8 (220 mg, 0.1 mmol) and 15 mL of chloroform were added to a reaction flask. 1 mL of trimethylamine (2 M) THF solution was added to the reaction mixture, and the mixture was stirred at 55 °C for 16 h. After the reaction was complete, the crude product precipitated and adhered to the bottom of the flask. The supernatant was discarded, and the solid precipitate was washed five times with chloroform. The solid precipitate was dissolved in 15 mL of methanol. 0.71 mL of trimethylamine (4.2 M) methanol solution was added to the solution, and the mixture was stirred at 55 °C for 16 h. The solution was then evaporated to dryness under vacuum. The final product was a purple solid (205 mg, 98%).

[0112] The proton NMR spectrum of compound 9 is shown below. Figure 2 As shown; the UV-Vis absorption spectrum of compound 9 in dimethyl sulfoxide solution is as follows. Figure 3 As shown: its maximum absorption wavelength is 413 nm, Q-band absorption peaks are 504, 542, 579, and 632 nm, its full width at half maximum (FWHM) is 20 nm, and its optical band gap is 2.23 eV; the fluorescence spectrum of compound 9 in dimethyl sulfoxide solution is shown below. Figure 4 As shown: λ(nm): 637,697;

[0113] 1 H NMR (400MHz, DMSO): δH=10.72(s,2H),9.74(d,J=4.6Hz,4H),9.19(d,J=4.5Hz,4H),8. 45(d,J=7.8Hz,4H),8.28(d,J=7.9Hz,4H),8.16(d,J=8.1Hz,4H),7.99(d,J=8.0Hz,4H ),7.10(d,J=2.8Hz,4H),4.18-4.16(m,8H),3.99-3.96(m,4H),3.99-3.96(m,12H),3. 08(s,54H),1.85-1.72(m,24H),1.61-1.52(m,12H),1.41-1.3(m,12H),-3.16(s,2H).

[0114] Example 3

[0115]

[0116] Synthesis of S2-3 and compound 10 (a cell membrane biomimetic conjugated electrolyte molecule 3 with a porphyrin unit as its core):

[0117] Compound 9 (200 mg, 0.12 mmol) was dissolved in 40 mL of methanol / chloroform solution (methanol:chloroform = 1:1), and Co(CH3COO)2·4H2O (290 mg, 1.2 mmol) was added. The mixture was heated to 40 °C under nitrogen protection and reacted for 12 h. After the solvent was evaporated to dryness, the product was reconstituted with 20 mL of deionized water and dialyzed against the aqueous solution for 24 h using a 20 kDa dialysis membrane to remove the metal salt. The product was then lyophilized and collected as a red powder (196 mg, 95%).

[0118] The UV-Vis absorption spectrum of compound 10 in dimethyl sulfoxide solution is as follows: Figure 3 As shown: its maximum absorption wavelength is 409 nm, the Q-band absorption peak is 522 nm, its full width at half maximum (FWHM) is 30 nm, and its optical band gap is 2.15 eV; the fluorescence spectrum of compound 10 in dimethyl sulfoxide solution is as follows. Figure 4 As shown: λ(nm): 596,637,697.

[0119] Example 4

[0120]

[0121] Synthesis of S2-3 and compound 11 (a cell membrane biomimetic conjugated electrolyte molecule 4 with a porphyrin unit as its core):

[0122] Compound 9 (200 mg, 0.12 mmol) was dissolved in 40 mL of methanol / chloroform solution (methanol:chloroform = 1:1), and Ni(CH3COO)2·4H2O (300 mg, 1.2 mmol) was added. The mixture was heated to 40 °C under nitrogen protection and reacted for 12 h. After the solvent was evaporated to dryness, the product was reconstituted with 20 mL of deionized water and dialyzed against the aqueous solution for 24 h using a 20 kDa dialysis membrane to remove the metal salt. The product was then lyophilized and collected as a red powder (196 mg, 95%).

[0123] The UV-Vis absorption spectrum of compound 11 in dimethyl sulfoxide solution is as follows: Figure 3 As shown: its maximum absorption wavelength is 410 nm, the Q-band absorption peaks are 516 and 548 nm, its full width at half maximum (FWHM) is 26 nm, and its optical band gap is 2.2 eV; the fluorescence spectrum of compound 11 in dimethyl sulfoxide solution is shown below. Figure 4 As shown: λ(nm): 596,637,697.

[0124] Comparative Example 1

[0125] A conjugated molecule, the structural formula of which is shown below:

[0126]

[0127] The UV-Vis absorption and fluorescence spectra of the conjugated molecule in dimethyl sulfoxide solution of Comparative Example 1 are as follows: Figure 5 , Figure 6 As shown. Its maximum absorption wavelength is 527 nm, and its full width at half maximum (FWHM) is 136 nm, which is much higher than that of the example. The optical band gap is 2.0 eV. The fluorescence spectrum of compound 11 in dimethyl sulfoxide solution is shown below. Figure 4 As shown: λ(nm): 604nm.

[0128] Test Example 1

[0129] Calcein test: In order to study the mechanical effect of the membrane biomimetic conjugated electrolyte molecules of the present invention on the lipid bilayer, we used a calcein leakage experiment and used an artificial membrane model (large monolayer vesicles, LUVs) to evaluate the integrity of the lipid membrane.

[0130] Specifically, calcein is a fluorescent dye. When encapsulated within liposomes, its fluorescence signal is weak due to a self-quenching effect. However, if the liposome membrane is damaged or its permeability increases, calcein leaks into the external solution, reducing its concentration and weakening the self-quenching effect, thus leading to a significant increase in fluorescence signal. Therefore, by monitoring changes in fluorescence intensity, the degree of perturbation of the lipid membrane by membrane-inspired molecules can be quantitatively assessed.

[0131] In sample vials, POPE (phosphatidylethanolamine) and POPG (phosphatidylglycerol) dissolved in chloroform were mixed at a molar ratio of 25:75 to obtain a 10 mM liposome solution. Methanol solutions of the conjugated electrolyte molecules from Examples 1, 2, and Comparative Example 1 were added to the liposome solution, with a molar ratio of liposomes to conjugated electrolyte molecules of 1000:5. The solution was then dried under an argon flow. To ensure complete solvent removal, the dried lipids were further dried overnight under vacuum, forming a thin lipid film inside the sample vial. The liposomes were rehydrated by adding a saturated calcein-containing PBS solution, then incubated at 65°C for 2 h, and the vesicles were squeezed through a 0.1 μm membrane 21 times with continuous stirring at approximately 300 rpm. Unencapsulated calcein was removed by dialyzing in aqueous solution for 3 days to prepare a 10 mM LUV sample. The liposomes were exposed to 20 mW cm⁻¹. -2 Under white light, the changes in its fluorescence intensity were monitored.

[0132] Test results are as follows Figure 8 As shown, compared with Comparative Example 1, after being exposed to light, the fluorescence intensity of Examples 1 and 2 changed less, indicating that the mechanical perturbation of the lipid membrane by the biomimetic conjugated electrolyte molecules designed in Examples 1 and 2 under light conditions was significantly lower than that in Comparative Example 1.

[0133] Test Example 2

[0134] Microbial phototoxicity experiment: In order to investigate the phototoxic effects of membrane-inspired conjugated electrolyte molecules on microorganisms, we evaluated their phototoxic effects by co-culturing Shewanella with different molecules (Example 1, Example 2 and Comparative Example 1) under white light irradiation.

[0135] Examples 1, 2, and Comparative Example 1, each at a concentration of 5 μM, were co-cultured with Shewanella bacteria (initial OD = 0.1) in the dark for 20 min to ensure cell membrane entry. They were then exposed to a 1600 LUX white light, and changes in OD values ​​were monitored every 2 hours. The test results are as follows: Figure 9 As shown, Examples 1 and 2 have less impact on microbial proliferation compared to Comparative Example 1 because Examples 1 and 2 introduce porphyrin units, whose macrocyclic structure enhances the rigidity of the probe and effectively increases the steric hindrance between the probe and phospholipid molecules, significantly restricting the free rotation of molecules under light conditions.

[0136] Among them, the confocal micrographs of cells in microorganisms in Examples 1 and 2 are as follows: Figure 7 As shown, this indicates that the molecules have been embedded in the cell membrane.

[0137] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, it is intended that all variations falling within the meaning and scope of equivalents of the claims be included within the present invention.

[0138] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit as its core, characterized in that, The cell membrane biomimetic conjugated electrolyte molecule with porphyrin unit as its core has the following structure: in, The Ar is selected from electron-donating groups having an aromatic ring structure; The —R1 + Selected from one or more alkyl derivatives or aryl derivatives containing quaternary ammonium salts or sulfonates as terminal groups; The M is absent, or is selected from metal atoms; X - Selected from —R1 + The oppositely charged ion of the group can react with -R1 + Groups form ion pairs; All carbon atoms on the alkyl derivative are not substituted, or one or more carbon atoms are substituted by one or more oxygen atoms, amino, sulfone, carbonyl, aryl, alkenyl, alkynyl, ester, cyano, or nitro groups. or All hydrogen atoms on the alkyl derivative are not substituted, or one or more hydrogen atoms are substituted by one or more of halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, or alkyne groups. The aryl derivative is selected from groups containing one or more benzene ring structures, wherein all hydrogen atoms on the benzene ring are not substituted; or One or more hydrogen atoms at any position on one or more benzene rings are substituted by one or more of a halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, alkyneic, carboxyl, ester, cyano, or nitro group.

2. The cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core according to claim 1, characterized in that, The —R1 + Selected from one or more alkyl groups containing quaternary ammonium salts or sulfonates as end groups; The number of repeating units of the alkyl group is 1-10.

3. The cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core according to claim 1, characterized in that, The Ar is selected from one or more aromatic monocyclic rings, or aromatic fused rings, and combinations thereof.

4. The cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core according to claim 1, characterized in that, The Ar is selected from one or more of the following structures connected together: in, X1 is selected independently from S, O, and Se; A is selected from carbon or silicon atoms; R2 is selected from aryl derivatives, alkyl derivatives, or hydrogen atoms; All carbon atoms on the alkyl derivative are not substituted, or one or more carbon atoms are substituted by one or more oxygen atoms, nitrogen atoms, amino groups, sulfone groups, carbonyl groups, aryl groups, alkenyl groups, alkynyl groups, ester groups, cyano groups, and nitro groups. or All hydrogen atoms on the alkyl derivative are not substituted, or one or more hydrogen atoms are substituted by one or more of halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, or alkyne groups. The aryl derivative is selected from groups containing one or more benzene ring structures, wherein all hydrogen atoms on the benzene ring are unsubstituted. or One or more hydrogen atoms at any position on one or more benzene rings are substituted by one or more of a halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, alkyneic, carboxyl, ester, cyano, or nitro group.

5. The cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core according to claim 1, characterized in that, The X - It is selected from one or more of the following: boron-containing organic anions, halogen anions, sulfur-containing organic anions, and acid radical anions.

6. The cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core according to claim 1, characterized in that, The M is absent or selected from transition metal atoms.

7. The method for preparing the cell membrane biomimetic conjugated electrolyte molecule with porphyrin unit as the core according to any one of claims 1-6, characterized in that, Includes the following steps: S1. Under the protection of an inert gas, the metal porphyrin containing Ar units and compound 5 are coupled together in the presence of a catalyst to obtain compound 6. S2. Compound 6 is treated to introduce ionic groups to obtain either cell membrane biomimetic conjugated electrolyte molecule 1 with porphyrin unit as the core or cell membrane biomimetic conjugated electrolyte molecule 2 with porphyrin unit as the core. Compound 5 is selected from the following structures: The -R3 is selected from one or more alkyl derivatives and aryl derivatives; X2 is selected from halogens; All carbon atoms on the alkyl derivative are not substituted, or one or more carbon atoms are substituted by one or more oxygen atoms, amino, sulfone, carbonyl, aryl, alkenyl, alkynyl, ester, cyano, or nitro groups. or All hydrogen atoms on the alkyl derivative are not substituted, or one or more hydrogen atoms are substituted by one or more of halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, or alkyne groups. The aryl derivative is selected from groups containing one or more benzene ring structures, wherein all hydrogen atoms on the benzene ring are not substituted; or One or more hydrogen atoms at any position on one or more benzene rings are substituted by one or more of a halogen, hydroxyl, amino, carboxyl, cyano, nitro, aryl, olefinic, alkyneic, carboxyl, ester, cyano, or nitro group.

8. The method for preparing a cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core according to claim 7, characterized in that, The coupling reaction is carried out at a temperature of 70-80°C; the ionic groups include one or more of the following: quaternary ammonium salt groups, phosphate groups, sulfonate groups, acetate groups, quaternary phosphate groups, phosphate ester groups, and amino groups.

9. The application of the cell membrane biomimetic conjugated electrolyte molecule with porphyrin unit core as described in any one of claims 1-6 in fluorescent probes, biosynthesis and catalysis.

10. The application according to claim 9, characterized in that, The fluorescent probe comprises liposomes and a cell membrane biomimetic conjugated electrolyte molecule with a porphyrin unit core; The molar ratio of the liposomes to the cell membrane biomimetic conjugated electrolyte molecules with porphyrin units as the core is 1000:5-20.

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