Method and system for evaluating luminescence property of near-infrared two-region molecular fluorescent probe

Abstract

The invention provides a method and a system for evaluating the luminescence performance of a near-infrared two-region molecular fluorescent probe, and relates to the technical field of fluorescent probes, the method comprises the following steps: obtaining the initial configuration of the near-infrared two-region molecular fluorescent probe in an aqueous solution, and carrying out molecular dynamics simulation on the initial configuration to obtain a plurality of representative conformations of the probe in the aqueous solution; performing quantum chemical calculation on each representative conformation to obtain first system energy of each representative conformation in a stable state and second system energy and intensity parameters of each representative conformation in a lowest singlet excitation state; generating a comprehensive evaluation factor based on the first system energy, the second system energy and the intensity parameter; and sorting the luminescence properties of the probes based on the comprehensive evaluation factor, and screening out the probe with the optimal performance. According to the method provided by the invention, quantitative evaluation on the luminescence property of the near-infrared two-region molecular fluorescent probe is realized, and technical support is provided for optimal design and clinical application of the probe.

Description

Technical Field

[0001] The present invention relates to the technical field of fluorescent probes, and specifically relates to a method and system for evaluating the luminescence performance of near-infrared II region molecular fluorescent probes. Background Art

[0002] The statements in this section merely provide background technologies related to the present disclosure and do not necessarily constitute prior art.

[0003] Due to advantages such as non-invasiveness, high sensitivity, and real-time dynamic monitoring, fluorescence imaging technology has been widely used in tumor detection, drug metabolism tracking, and surgical navigation in biomedical research. Traditional visible light region (400 - 700 nm) and near-infrared I region (NIR-I, 700 - 900 nm) fluorescent probes are limited in imaging depth and signal-to-noise ratio due to the high scattering and autofluorescence interference of biological tissues. In contrast, NIR-II fluorescent probes utilize photons with longer wavelengths (1000 - 1700 nm), significantly reducing scattering and background noise, and improving penetration depth and resolution, becoming a research hotspot in biological in vivo imaging.

[0004] Currently, the evaluation of NIR-II probes mainly relies on experimental tests, including spectroscopic analysis methods and in vitro cell experiments. Among them, spectroscopic analysis methods measure fluorescence intensity, lifetime, and quantum yield, but ignore the influence of molecular dynamic conformations; in vitro cell experiments verify the probe performance through cell imaging, but the experimental conditions are difficult to standardize. How to scientifically and systematically evaluate the luminescence performance of NIR-II probes has become an urgent problem to be solved currently. Summary of the Invention

[0005] To solve the above problems, the present invention provides a method and system for evaluating the luminescence performance of near-infrared II region molecular fluorescent probes. By integrating molecular dynamics simulation and quantum chemical calculation, a multi-dimensional evaluation factor is constructed to solve the problems of single evaluation index, inconsistent standards, and high experimental costs in the prior art.

[0006] The first aspect of the present invention provides a method for evaluating the luminescence performance of near-infrared II region molecular fluorescent probes, including: S1: Obtain the initial conformation of the near-infrared II region molecular fluorescent probe in aqueous solution, perform molecular dynamics simulation on the initial conformation to obtain multiple representative conformations of the probe in aqueous solution; S2: Perform quantum chemical calculation on each representative conformation to obtain the first system energy of each representative conformation in the stable state, the second system energy and intensity parameters in the lowest singlet excited state; S3: Generate a comprehensive evaluation factor based on the first system energy, the second system energy, and the intensity parameters; S4: Rank the luminescence performance of the probes based on the comprehensive evaluation factor and screen out the probe with the best performance.

[0007] Further, in S1, performing molecular dynamics simulation on the initial configuration specifically includes: S1.1: Constructing a molecular structure model of the probe based on the initial configuration, converting the molecular structure model into a format compatible with molecular dynamics simulation software, and generating a topology file suitable for molecular force field description; S1.2: Defining the three-dimensional space boundary of the simulation system, adding water molecules thereto to form a simulation environment including the probe molecule and water molecules; S1.3: Eliminating the high-energy configurations of the system through energy minimization, and then performing molecular dynamics simulation under set time conditions, and screening to obtain multiple representative conformations of the probe in water molecules by using the clustering analysis method.

[0008] Further, the clustering analysis method screens conformations by setting a cut-off value and retains the local environment information of the probe molecule and the water molecules around it.

[0009] Further, S2 includes: S2.1: Based on the density functional method and the ONIOM model, dividing the probe molecule and its solvent environment into a high layer and a low layer, respectively describing them with a high-precision quantum chemical theory model and a simplified force field model, and calculating the first system energy of each representative conformation in the stable state through structure optimization; S2.2: Based on the time-dependent density functional method and the ONIOM model, dividing the probe molecule and its solvent environment into a high layer and a low layer, respectively describing them with a high-precision quantum chemical theory model and a simplified force field model, calculating the second system energy of each representative conformation in the lowest singlet excited state through excited state structure optimization, and simultaneously obtaining the oscillator strength, i.e., the intensity parameter, in the lowest singlet excited state.

[0010] Further, S3 includes: S3.1: Calculating the first system energy E a and the second system energy E b the energy difference E ; S3.2: Performing correlation analysis on the energy difference E and the intensity parameter I to generate a comprehensive evaluation factor X .

[0011] Further, S3.2 includes: S3.2.1: Statistically analyzing n the intensity parameters I 1 , I 2 ,I 3 ... I n and energy parameters E 1 , E 2 , E 3 ... E n ; S3.2.2: Construct the intensity matrix and the energy matrix whose matrix elements are respectively and where ; S3.2.3: Add the matrices I and E to obtain the matrix with matrix element , ; S3.2.4: Use the row sum of each row of the matrix C as the minuend and 2 n as the subtrahend to calculate the difference to obtain the correction factor , ; S3.2.5: Generate a comprehensive evaluation factor based on the above data.

[0012] Furthermore, the S4 includes: judging the luminescence performance of the probe based on the value of the comprehensive evaluation factor. The larger the value, the better the luminescence performance. The molecular fluorescence probe with the largest value is the probe with the best performance.

[0013] The second aspect of the present invention provides a system for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe, including: A conformation acquisition unit for acquiring the initial conformation of the near-infrared second-region molecular fluorescence probe in an aqueous solution, performing molecular dynamics simulation on the initial conformation to obtain multiple representative conformations of the probe in the aqueous solution; An evaluation parameter acquisition unit for performing quantum chemical calculations on each representative conformation to obtain the first system energy of each representative conformation in the stable state, the second system energy in the lowest singlet excited state, and the intensity parameter; An evaluation factor calculation unit for generating a comprehensive evaluation factor based on the first system energy, the second system energy, and the intensity parameter; An evaluation and screening unit for ranking the luminescence performance of the probe based on the comprehensive evaluation factor and screening out the probe with the best performance.

[0014] The third aspect of the present invention provides an evaluation device for the luminescence performance of a near-infrared second-region molecular fluorescence probe. The device includes a memory and a processor; the memory is used to store a computer program; the processor is used to implement the above-mentioned method for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe when executing the computer program.

[0015] The fourth aspect of the present invention provides a computer-readable storage medium, on which a computer program is stored. When the computer program is executed by a processor, the above-mentioned method for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe is implemented.

[0016] Compared with the prior art, the method and system for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe provided by the present invention have the following beneficial effects: Based on the structural characteristics of the near-infrared second-region molecular fluorescence probe, the present invention constructs a comprehensive luminescence performance evaluation index by comprehensively considering influencing factors in multiple dimensions and introducing the weight of the time evolution effect. Using this index, the luminescence performance of the near-infrared second-region molecular fluorescence probe can be quantitatively considered under a unified standard. The method proposed by the present invention has the advantages of low cost, environmental friendliness, and high reliability, and can greatly save the input costs of time and resources. By applying the method of the present invention, the evaluation process of the luminescence performance of the near-infrared second-region molecular fluorescence probe can be effectively accelerated, providing technical support for the screening of high-performance near-infrared second-region molecular fluorescence probes, and laying a solid theoretical foundation for the design of new near-infrared second-region molecular fluorescence probes. The present invention not only provides a strong tool support for scientific research and medical applications, but also promotes the technological innovation and development in the field of biofluorescence imaging. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The accompanying drawings forming a part of this disclosure are used to provide a further understanding of this disclosure. The schematic embodiments and descriptions thereof of this disclosure are used to explain this disclosure and do not constitute an improper limitation of this disclosure.

[0018] Figure 1 is a block diagram of the steps of the method for evaluating the probe luminescence performance provided by the present invention; Figure 2 is a flowchart of the steps of the method for evaluating the probe luminescence performance provided by the present invention; Figure 3 is a schematic structural diagram of Fluo1 provided by the present invention; Figure 4 is a schematic diagram of the representative conformation 1 of Fluo1 in an aqueous solution provided by the present invention; Figure 5 is a schematic diagram of the representative conformation 2 of Fluo1 in an aqueous solution provided by the present invention; Figure 6It is a schematic diagram of the representative conformation 3 of Fluo1 in aqueous solution provided by the present invention; Figure 7 It is a schematic diagram of the representative conformation 4 of Fluo1 in aqueous solution provided by the present invention; Figure 8 It is a schematic diagram of the representative conformation 5 of Fluo1 in aqueous solution provided by the present invention; Figure 9 It is a schematic diagram of the representative conformation 6 of Fluo1 in aqueous solution provided by the present invention; Figure 10 It is a schematic diagram of the representative conformation 7 of Fluo1 in aqueous solution provided by the present invention; Figure 11 It is a schematic diagram of the structure of Fluo2 provided by the present invention; Figure 12 It is a schematic diagram of the structure of Fluo3 provided by the present invention; Figure 13 It is a schematic diagram of the representative conformation 1 of Fluo2 in aqueous solution provided by the present invention; Figure 14 It is a schematic diagram of the representative conformation 2 of Fluo2 in aqueous solution provided by the present invention; Figure 15 It is a schematic diagram of the representative conformation 3 of Fluo2 in aqueous solution provided by the present invention; Figure 16 It is a schematic diagram of the representative conformation 4 of Fluo2 in aqueous solution provided by the present invention; Figure 17 It is a schematic diagram of the representative conformation 1 of Fluo3 in aqueous solution provided by the present invention; Figure 18 It is a schematic diagram of the representative conformation 2 of Fluo3 in aqueous solution provided by the present invention; Figure 19 It is a schematic diagram of the representative conformation 3 of Fluo3 in aqueous solution provided by the present invention; Figure 20 It is a schematic diagram of the representative conformation 4 of Fluo3 in aqueous solution provided by the present invention; Figure 21 It is a schematic diagram of the probe luminescence performance evaluation system provided by the present invention. Detailed implementation manners

[0019] It should be noted that the following detailed description is exemplary and is intended to provide further illustration of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present invention belongs.

[0020] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular forms are also intended to include the plural forms. In addition, it should be understood that the terms "comprising" and "having" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product or device comprising a series of steps or units need not be limited to those steps or units clearly listed, but may include other steps or units not clearly listed or inherent to these processes, methods, products or devices.

[0021] In the case of no conflict, the embodiments in the present invention and the features in the embodiments may be combined with each other.

[0022] All data acquisition in this embodiment is based on compliance with laws, regulations and user consent, and is a legal application of the data.

[0023] Embodiment 1 Please refer to the attached specification Figure 1 and Figure 2 , Figure 1 is a block diagram of the steps of the method for evaluating the luminescence performance of the probe provided by the present invention, Figure 2 is a flowchart of the steps of the method for evaluating the luminescence performance of the probe provided by the present invention. From Figure 1 and Figure 2 , it can be seen that the method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe includes: S1: Obtain the initial configuration of the near-infrared second-region molecular fluorescence probe in aqueous solution, perform molecular dynamics simulation on the initial configuration, and obtain multiple representative conformations of the probe in aqueous solution; S2: Perform quantum chemical calculations on each representative conformation to obtain the first system energy in the stable state, the second system energy in the lowest singlet excited state, and the intensity parameter of each representative conformation; S3: Generate a comprehensive evaluation factor based on the first system energy, the second system energy, and the intensity parameter; S4: Rank the luminescence performance of the probe based on the comprehensive evaluation factor, and screen out the probe with the best performance.

[0024] Specifically, the molecular dynamics simulation of the initial configuration in S1 specifically includes: S1.1: Construct a molecular structure model of the probe based on the initial configuration, convert the molecular structure model into a format compatible with the molecular dynamics simulation software, and generate a topology file suitable for molecular force field description; S1.2: Define the three-dimensional space boundary of the simulation system, add water molecules to it, and form a simulation environment including the probe molecule and water molecules; S1.3: Eliminate the high-energy configurations of the system through energy minimization, and then perform molecular dynamics simulations under set time conditions. Use the clustering analysis method to screen and obtain multiple representative conformations of the probe in water molecules.

[0025] Specifically, the clustering analysis method screens conformations by setting a cut-off value and retains the local environmental information of the probe molecule and the water molecules around it.

[0026] In a specific embodiment, molecular dynamics simulations are performed based on the initial configuration of the NIR-II molecular fluorescence probe in an aqueous solution, specifically including: (1) Construct the structural model of the NIR-II molecular fluorescence probe in ChemDraw software, and use AmberTools to convert it into the.gro format, and simultaneously generate its topological file under the GAFF force field; (2) Define the simulation box in GROMACS software and add water solvent to the box. The preferred box size is 6 nm × 6 nm × 6 nm, which includes 1 NIR-II molecular fluorescence probe and thousands of water molecules; (3) Run energy minimization calculations and 20 ns of kinetic simulations through GROMACS. The preferred simulation time step is 2 fs, and the temperature is set at 298.15 K.

[0027] In S1, the clustering analysis method is used to obtain the n representative conformations of the NIR-II molecular fluorescence probe in an aqueous solution. The preferred cutoff value does not exceed 0.1 Å. Screen out the intermediate structures in the clusters with larger capacities, and select the water molecules within 5 Å of the NIR-II molecular fluorescence probe and its surroundings as representative conformations for subsequent calculations. n

[0028] Specifically, S2 includes: S2.1: Based on the density functional method and the ONIOM model, divide the probe molecule and its solvent environment into a high layer and a low layer, and describe them with a high-precision quantum chemical theory model and a simplified force field model respectively. Calculate the first system energy of each representative conformation in the stable state through structural optimization; S2.2: Based on the time-dependent density functional method and the ONIOM model, divide the probe molecule and its solvent environment into a high layer and a low layer, and describe them with a high-precision quantum chemical theory model and a simplified force field model respectively. Calculate the second system energy of each representative conformation in the lowest singlet excited state through excited state structural optimization, and simultaneously obtain the oscillator strength, that is, the intensity parameter, in the lowest singlet excited state.

[0029] In a specific embodiment, the first system energy of each representative conformation in the stable state ( E a ), the second system energy in the lowest singlet excited state ( E b ), and the intensity parameter ( I ) are obtained through quantum chemical calculations, specifically including: (1) Based on the density functional method and the ONIOM model, the NIR-II molecular fluorescence probe and the surrounding water molecules are respectively set as the high layer and the low layer, and are described by the cam-b3lyp / 6-31g(d) and UFF force fields respectively. The energy corresponding to the structure of each representative conformation in the stable state, that is, the first system energy, is obtained through structure optimization in the GAUSSIAN 16 software E a ; (2) Based on the time-dependent density functional method and the ONIOM model, the NIR-II molecular fluorescence probe and the surrounding water molecules are respectively set as the high layer and the low layer, and are described by the cam-b3lyp / 6-31g(d) and UFF force fields respectively. The energy corresponding to the structure of each representative conformation in the lowest singlet excited state, that is, the second system energy, is obtained through the lowest singlet excited state structure optimization in the GAUSSIAN 16 software E b , and at the same time, the oscillator strength of the lowest singlet excited state, that is, the intensity parameter, is obtained I .

[0030] Specifically, S3 includes: S3.1: Calculate the energy difference E a between the first system energy E b and the second system energy E， ; S3.2: Perform correlation analysis on the energy difference E and the intensity parameter I to generate a comprehensive evaluation factor X through mathematical modeling.

[0031] Specifically, S3.2 includes: S3.2.1: Statistically analyze n the intensity parameters I 1 , I 2 , I 3 ... I n and the energy parameters E 1 ofE 2 , E 3 ... E n ; S3.2.2: Construct the intensity matrix and the energy matrix , and their matrix elements are respectively and , where ; S3.2.3: Add the matrices I and E to obtain the matrix , and the matrix element , ; S3.2.4: Use the row sum of each row of the matrix C as the minuend, and 2 n as the subtrahend, and take the difference to obtain the correction factor , ; S3.2.5: Generate the comprehensive evaluation factor .

[0032] Specifically, S4 includes: judging the luminescence performance of the probe based on the numerical value of the comprehensive evaluation factor. The larger the numerical value, the better the luminescence performance. The molecular fluorescence probe with the largest numerical value is the probe with the best performance.

[0033] In a specific embodiment, the NIR-II molecular fluorescence probe is named Fluo1, and the structure of Fluo1 can be known from Figure 3 .

[0034] Step 1: Perform molecular dynamics simulations based on the initial conformation of Fluo1 in aqueous solution to obtain 7 representative conformations of Fluo1 in the aqueous solution environment, specifically including: (1) Construct the structural model of Fluo1 in ChemDraw software (as shown in Figure 3 ), and use AmberTools to convert it into the.gro format, and generate its topology file under the GAFF force field at the same time; (2) Define a simulation box with a size of 6 nm × 6 nm × 6 nm in GROMACS software, and add water solvent to the box. The box contains 1 Fluo1 and 6994 water molecules; (3) Run energy minimization calculations and 20 ns of molecular dynamics simulations through GROMACS. The simulation time step is 2 fs, and the temperature is set to 298.15 K.

[0035] (4)Based on the results of kinetic simulations, a cutoff value of 0.1 Å was set for clustering analysis. The intermediate structures in the 7 clusters with larger capacities were screened out, and the water molecules containing Fluo1 and within a range of 5 Å around it were selected as representative conformations (as Figures 4 to 10 shown), for subsequent calculations.

[0036] Step 2: Obtain the first system energy of each representative conformation at the steady state ( E a ), the second system energy at the lowest singlet excited state ( E b ), and the intensity parameter ( I ), specifically including: (1) Based on the density functional method and the ONIOM model, Fluo1 and the surrounding water molecules were respectively set as the high layer and the low layer, and were described by the cam-b3lyp / 6-31g(d) and UFF force fields respectively. The energy corresponding to the structure of each representative conformation at the steady state, that is, the first system energy, was obtained through structural optimization in the GAUSSIAN 16 software E a (see Table 1); (2) Based on the time-dependent density functional method and the ONIOM model, Fluo1 and the surrounding water molecules were respectively set as the high layer and the low layer, and were described by the cam-b3lyp / 6-31g(d) and UFF force fields respectively. The energy corresponding to the structure of each representative conformation at the lowest singlet excited state, that is, the second system energy, was obtained through structural optimization of the lowest singlet excited state in the GAUSSIAN 16 software E b (see Table 1), and at the same time, the oscillator strength of the lowest singlet excited state, that is, the intensity parameter, was obtained I (see Table 1); Table 1 The first system energy of the representative conformations of Fluo1 E a (unit: eV), the second system energy E b (unit: eV), the energy parameter E (unit: eV), and the intensity parameter I (unit: a.u.)

[0037] Step 3: Calculate the evaluation factors based on the obtained first system energy ( E a ), second system energy ( E b ), and intensity parameter ( I ), specifically including: (1) Calculate the energy parameter according to the first system energy and the second system energy of each representative conformation E a and the second system energy E b Calculate the energy parameter (see Table 1); (2) Statistically analyze the intensity parameters of 7 representative conformations I 1 = 0.452, I 2 = 0.571, I 3 = 0.529, I 4 = 0.398, I 5 = 0.414 ，I 6 = 0.456 ，I 7 = 0.496 and the energy parameter E 1 = 1.978, E 2 = 2.152, E 3 = 2.701, E 4 = 2.887, E 5 = 2.309 ，E 6 = 2.165 ，E 7 = 2.435; (3) Construct the intensity matrix with as matrix elements, and construct the energy matrix with as matrix elements; Construct the intensity matrix with as matrix elements, and construct the energy matrix with as matrix elements; and Construct the energy matrix with as matrix elements; ; (4) Add the matrices and to get; I and E Add to get ; (5) Use the row sum of each row of the matrix as the minuend, and use 2 C Each row sum of the matrix is used as the minuend, and 2× Taking 7 as the minuend, the difference is used as the correction factor

[0038] (6)Calculating the evaluation factor of Fluo1 based on the above data 。

[0039] Step 4: Judging the luminescence performance of the NIR-II molecular fluorescence probe through the evaluation factor, specifically: the luminescence performance evaluation factor of Fluo1 is 1.174

[0040] In a specific embodiment, two NIR-II molecular fluorescence probes are named Fluo2 and Fluo3 respectively. From Figure 11 the structure of Fluo2 can be known, and from Figure 12 the structure of Fluo3 can be known

[0041] Step 1: Based on the initial configurations of Fluo2 and Fluo3 in aqueous solution, perform molecular dynamics simulations to obtain 4 representative conformations of Fluo2 and Fluo3 in the aqueous solution environment respectively, as shown in Figures 13 to 16 and Figures 17 to 20 respectively, specifically including: (1)Construct the structural models of Fluo2 and Fluo3 in ChemDraw software (as shown in Figure 11 and Figure 12 respectively), and use AmberTools to convert them into the.gro format, and at the same time generate the topology files of Fluo2 and Fluo3 under the GAFF force field (2)Define a simulation box with a size of 6 nm × 6 nm × 6 nm in GROMACS software, and add water solvent to the box. The box contains 1 Fluo2 and 6994 water molecules, or 1 Fluo3 and 6990 water molecules (3)Run energy minimization calculations and 20 ns of kinetic simulations through GROMACS. The simulation time step is 2 fs, and the temperature is set to 298.15 K

[0042] (4)Based on the kinetic simulation results of Fluo2 and Fluo3, set the cutoff value to 0.03 Å for clustering analysis, screen out the intermediate structures in the 4 clusters with larger capacities respectively, and select the water molecules within 5 Å around Fluo2 (or Fluo3) and itself as representative conformations for subsequent calculations

[0043] Step 2: Obtain the first system energy of each representative conformation at the steady state through quantum chemical calculations ( E a), the second system energy in the lowest singlet excited state ( E b ), and the intensity parameter ( I ), specifically including: (1) Based on the density functional method and the ONIOM model, Fluo2 (or Fluo3) and the surrounding water molecules are set as the high layer and the low layer respectively, and are described by the cam-b3lyp / 6-31g(d) and UFF force fields respectively. The energy corresponding to the structure of each representative conformation in the stable state is obtained through structural optimization in the GAUSSIAN 16 software, which is the first system energy E a (see Table 2); (2) Based on the time-dependent density functional method and the ONIOM model, Fluo2 (or Fluo3) and the surrounding water molecules are set as the high layer and the low layer respectively, and are described by the cam-b3lyp / 6-31g(d) and UFF force fields respectively. The energy corresponding to the structure of each representative conformation in the lowest singlet excited state is obtained through the lowest singlet excited state structural optimization in the GAUSSIAN 16 software, which is the second system energy E b (see Table 2). At the same time, the oscillator strength in the lowest singlet excited state, that is, the intensity parameter I (see Table 2); Table 2 The first system energy of the representative conformations of Fluo2 and Fluo3 E a (unit: eV), the second system energy E b (unit: eV), the energy parameter E (unit: eV), and the intensity parameter I (unit: a.u.)

[0044] Step 3: Calculate the evaluation factors based on the obtained first system energy ( E a ), the second system energy ( E b ), and the intensity parameter ( I ), specifically including: (1) Calculate the energy parameter E a according to the first system energy E b and the second system energy of each representative conformation (see Table 2); (2) Statistically analyze the intensity parameter I 1 = 0.540 of each representative conformation of Fluo2,I 2 = 0.613, I 3 = 0.698, I 4 =0.566 and energy parameter E 1 = 2.482, E 2 = 2.304, E 3 = 2.270, E 4 = 1.531, while counting the intensity parameters of each representative conformation of Fluo3 I' 1 = 0.750, I' 2 = 0.612, I' 3 = 0.644, I' 4 = 0.676 and energy parameter E' 1 = 2.709, E' 2 = 1.676, E' 3 = 1.775, E' 4 = 2.675; (3) Using and as matrix elements to construct the intensity matrix of Fluo2 and energy matrix , while using and as matrix elements to construct the intensity matrix of Fluo3 and energy matrix .

[0045] (4) Adding the matrices I and E to obtain the matrix of Fluo2, while adding the matrices and to obtain the matrix of Fluo3

[0046] (5) Using the row sum of each row of the matrices C and as the minuend, and 2×4 as the subtrahend, and taking the difference to obtain the correction factor P 1=-0.06, P2 = -0.33, P 3 = -0.76, P 4 = 1.88 and P 1' = -1.16, P 2' = 1.64, P 3' = 1.16, P 4' = -0.74; (6) Calculate the evaluation factors of Fluo2 and Fluo3 based on the above data, which are respectively and .

[0047] Step 4: Determine the luminescence performance of the NIR-II molecular fluorescence probe through the evaluation factor, and screen out the probe with the optimal performance. Specifically, the luminescence performance evaluation factors of Fluo2 and Fluo3 are 1.272 and 1.330 respectively, indicating that Fluo3 has better luminescence performance. Therefore, compared with Fluo2, Fluo3 is the screened probe with the optimal luminescence performance.

[0048] Example 2 As Figure 21 shown, this example provides a system for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe, including: A conformation acquisition unit for acquiring the initial conformation of the near-infrared second-region molecular fluorescence probe in an aqueous solution, performing molecular dynamics simulation on the initial conformation, and obtaining multiple representative conformations of the probe in the aqueous solution; An evaluation parameter acquisition unit for performing quantum chemical calculations on each representative conformation to obtain the first system energy in the stable state, the second system energy in the lowest singlet excited state, and the intensity parameter of each representative conformation; An evaluation factor calculation unit for generating a comprehensive evaluation factor based on the first system energy, the second system energy, and the intensity parameter; An evaluation and screening unit for ranking the luminescence performance of the probe based on the comprehensive evaluation factor and screening out the probe with the optimal performance.

[0049] Example 3 This example provides a device for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe. The device includes a memory and a processor; the memory is used to store a computer program; the processor is used to implement the above method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe when executing the computer program.

[0050] Among them, the processor is connected to the memory, and the above one or more computer programs are stored in the memory. When the electronic device runs, the processor executes the one or more computer programs stored in the memory, so that the electronic device executes the method described in Example 1 above.

[0051] It should be understood that in this embodiment, the processor may be a central processing unit (CPU), and the processor may also be other general-purpose processors, digital signal processors (DSPs), application-specific integrated circuits (ASICs), field-programmable gate arrays (FPGAs), or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc. The general-purpose processor may be a microprocessor or the processor may also be any conventional processor, etc.

[0052] The memory may include a read-only memory and a random access memory, and provide instructions and data to the processor. A part of the memory may also include a non-volatile random access memory. For example, the memory may also store information about the device type.

[0053] In the implementation process, each step of the above method may be completed by the integrated logic circuit in the hardware of the processor or the instructions in the form of software.

[0054] The method in the first embodiment may be directly embodied as being executed and completed by the hardware processor, or executed and completed by the combination of the hardware and software modules in the processor. The software module may be located in a mature storage medium in the art such as a random access memory, a flash memory, a read-only memory, a programmable read-only memory, or an electrically erasable programmable memory, a register, etc. This storage medium is located in the memory, and the processor reads the information in the memory and combines its hardware to complete the steps of the above method. To avoid repetition, it will not be described in detail here.

[0055] Those of ordinary skill in the art can realize that the units and algorithm steps of each example described in combination with this embodiment can be implemented by electronic hardware or a combination of computer software and electronic hardware. Whether these functions are executed in a hardware or software manner depends on the specific application and design constraints of the technical solution. Professional technicians can use different methods to implement the described functions for each specific application, but this implementation should not be considered to exceed the scope of the present invention.

[0056] Embodiment 4 A computer-readable storage medium provided in another embodiment of the present invention stores a computer program, and when the computer program is executed by a processor, the method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe as described above is implemented.

[0057] Those of ordinary skill in the art can understand that all or part of the processes in the methods of the above embodiments can be completed by instructing relevant hardware through a computer program. The program can be stored in a computer-readable storage medium. When the program is executed, it can include the processes of the embodiments of the above methods. Among them, the storage medium can be a magnetic disk, an optical disk, a read-only memory (ROM), or a random access memory (RAM), etc. In this application, the units described as separate components may or may not be physically separated, and the components shown as units may or may not be physical units, that is, they can be located in one place or distributed to multiple network units. One can select some or all of the units according to actual needs to achieve the purpose of the solution of the embodiments of the present invention. In addition, the functional units in each embodiment of the present invention can be integrated into a processing unit, or each unit can exist physically alone, or two or more units can be integrated into one unit. The above integrated units can be implemented in the form of hardware or in the form of software functional units.

[0058] Although the present invention is disclosed as above, the protection scope of the present invention is not limited thereto. Without departing from the spirit and scope of the present invention, those skilled in the art can make various changes and modifications, and these changes and modifications will all fall within the protection scope of the present invention.

Claims

1. A method for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe, characterized in that, Including: S1: Obtain the initial configuration of the near-infrared second-region molecular fluorescence probe in aqueous solution, perform molecular dynamics simulations on the initial configuration, and obtain multiple representative conformations of the probe in aqueous solution; S2: Perform quantum chemical calculations on each representative conformation to obtain the first system energy of each representative conformation in the stable state, the second system energy and intensity parameters in the lowest singlet excited state; S3: Generate a comprehensive evaluation factor based on the first system energy, the second system energy and the intensity parameters; S4: Rank the luminescence performance of the probes based on the comprehensive evaluation factor, and screen out the probe with the best performance.

2. The method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to claim 1, wherein, In the S1, the molecular dynamics simulation of the initial configuration specifically includes: S1.1: Construct a molecular structure model of the probe based on the initial configuration, convert the molecular structure model into a format compatible with the molecular dynamics simulation software, and generate a topology file suitable for molecular force field description; S1.2: Define the three-dimensional space boundary of the simulation system, add water molecules to it to form a simulation environment including the probe molecule and water molecules; S1.3: Eliminate the high-energy configurations of the system through energy minimization, and then perform molecular dynamics simulations under set time conditions. Use the clustering analysis method to screen out multiple representative conformations of the probe in water molecules.

3. The method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to claim 2, wherein The clustering analysis method screens conformations by setting a cut-off value and retains the local environment information of the probe molecule and the surrounding water molecules.

4. The method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to claim 1, wherein The S2 includes: S2.1: Based on the density functional method and the ONIOM model, divide the probe molecule and its solvent environment into a high layer and a low layer, and describe them with a high-precision quantum chemical theory model and a simplified force field model respectively. Calculate the first system energy of each representative conformation in the stable state through structure optimization; S2.2: Based on the time-dependent density functional method and the ONIOM model, divide the probe molecule and its solvent environment into a high layer and a low layer, and describe them with a high-precision quantum chemical theory model and a simplified force field model respectively. Calculate the second system energy of each representative conformation in the lowest singlet excited state through excited state structure optimization, and at the same time obtain the oscillator strength in the lowest singlet excited state, that is, the intensity parameter.

5. The method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to claim 1, wherein, The S3 includes: S3.1: Calculate the energy of the first system E a and the energy of the second system E b to obtain the energy difference E ; S3.2: Correlate the energy difference E with the intensity parameter I to conduct a correlation analysis and generate a comprehensive evaluation factor X through mathematical modeling.

6. The method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to claim 5, wherein The S3.2 includes: S3.2.1: Statistics n Intensity parameters of I 1 , I 2 , I 3 ... I n and energy parameters E 1 , E 2 , E 3 ... E n ; S3.2.2: Construct the strength matrix and the energy matrix , and their matrix elements are respectively and , where ; S3.2.3: Add the matrices I and E to obtain the matrix , with matrix elements , ; S3.2.4: Using the matrix C The row sum of each row as the minuend, and 2 n as the subtrahend, subtract to obtain the correction factor , ; S3.2.5: Generate comprehensive evaluation factors based on the above data .

7. The method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to claim 1, wherein The S4 includes: Judging the luminescence performance of the probe based on the numerical value of the comprehensive evaluation factor. The larger the numerical value, the better the luminescence performance. The molecular fluorescence probe with the largest numerical value is the probe with the best performance.

8. A system for evaluating the luminescence performance of a near-infrared second-region molecular fluorescence probe, characterized in that, Including: A conformation acquisition unit for obtaining the initial configuration of the near-infrared second-region molecular fluorescence probe in aqueous solution, performing molecular dynamics simulations on the initial configuration, and obtaining multiple representative conformations of the probe in aqueous solution; An evaluation parameter acquisition unit for performing quantum chemical calculations on each representative conformation to obtain the first system energy of each representative conformation in the stable state, the second system energy and intensity parameters in the lowest singlet excited state; An evaluation factor calculation unit for generating a comprehensive evaluation factor based on the first system energy, the second system energy and the intensity parameters; An evaluation and screening unit for ranking the luminescence performance of the probes based on the comprehensive evaluation factor, and screening out the probe with the best performance.

9. An evaluation device for the luminescence performance of a near-infrared second-region molecular fluorescence probe, characterized in that, The device includes a memory and a processor; the memory is used for storing a computer program; the processor is used for implementing the method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to any one of claims 1 to 7 when executing the computer program.

10. A computer-readable storage medium, characterized in that, A computer program is stored on the storage medium, and when the computer program is executed by a processor, the method for evaluating the luminescence performance of the near-infrared second-region molecular fluorescence probe according to any one of claims 1 to 7 is implemented.

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