Organic molecules, nanoparticles and their applications with ultrasound-activated luminescence properties

Abstract

The application discloses organic molecules, nanoparticles and applications with ultrasonic activated luminescence properties, the organic molecules including porphyrin molecules, cyanine molecules, BODIPY molecules, organic semiconductor polymer molecules containing thiophene structures, organic semiconductor polymer molecules containing styrene structures, anthracene derivatives. The application uses several kinds of organic molecules with ultrasonic activated luminescence as luminophores through molecular structure screening, uses amphiphilic polymers as surfactants, and obtains nanoparticles with ultrasonic activated luminescence through ultrasonic treatment. The application also provides applications of the ultrasonic activated luminescence system in in-vivo imaging, the system uses organic molecules with ultrasonic activated luminescence as luminophores, can construct an ultrasonic activated luminescence system with near-infrared emission, and realizes high-performance in-vivo imaging applications.

Description

Technical Field

[0001] This invention belongs to the field of luminescent materials, specifically relating to organic molecules, nanoparticles, and applications that possess ultrasonically activated luminescence properties. Background Technology

[0002] Molecular imaging is a non-invasive tool used for the detection, diagnosis, prediction, and monitoring of diseases. Optical imaging is a crucial component of molecular imaging, offering advantages such as high sensitivity, specificity, and real-time detection. Currently, molecules or materials primarily emit light through activation by lasers, X-rays, radionuclides, or chemical / biochemical interactions. In fluorescence imaging, real-time photoexcitation produces tissue autofluorescence, which reduces the sensitivity and specificity of in vivo imaging. Compared to fluorescence imaging, chemiluminescence, bioluminescence, Cherenkov luminescence, or X-ray activated luminescence imaging can eliminate the autofluorescence of biological tissues. However, bioluminescent or chemiluminescent signals are often affected by the enzyme microenvironment or substrate biodistribution in living animals. Cherenkov luminescence or X-ray activated luminescence imaging typically requires high doses of X-rays or radioisotopes to obtain sufficient imaging contrast, which may damage normal tissue.

[0003] Ultrasound imaging uses high-frequency waves to visualize the interior of the human body. Unlike X-rays, high-frequency ultrasound does not emit ionizing radiation. Furthermore, ultrasound imaging is a widely used, low-cost, real-time, non-invasive, and safe imaging method, with broad applications in clinical anatomy and functional imaging. Considering these advantages, ultrasound holds promise as an energy source for activating luminescence. Summary of the Invention

[0004] The purpose of this invention is to provide organic molecules, nanoparticles, and applications with ultrasound-activated luminescence properties to reduce imaging background, increase luminescence intensity, and enhance tissue penetration, thereby achieving high-performance in vivo imaging.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] This invention provides organic molecules with ultrasound-activated luminescence properties, including porphyrin molecules, anthocyanin molecules, BODIPY molecules, organic semiconductor polymer molecules, anthracene derivatives, etc., and their structural formulas include, but are not limited to:

[0007]

[0008]

[0009]

[0010] The present invention also provides nanoparticles with ultrasound-activated luminescence properties, said nanoparticles being made from the aforementioned organic molecules;

[0011] Alternatively, it can be made from the aforementioned organic molecules and the luminescence enhancer HBA; the luminescence intensity of the organic molecules is further enhanced by HBA, the structural formula of which is as follows:

[0012]

[0013] The nanoparticles are prepared and synthesized by a self-assembly method, with organic molecules as the core and amphiphilic polymers as the surface modification layer.

[0014] The nanoparticles have a uniform spherical structure; the size of the nanoparticles ranges from 1 to 1000 nanometers; the ultrasonic frequency range for exciting the nanoparticles is 30 kHz to 3 MHz; the emission wavelength range of the organic molecules or nanoparticles is 200 to 2000 nanometers; the luminescence time ranges from 1 second to 24 hours; and the luminescence intensity ranges from 10... 2 ~10 10 p / sec.

[0015] The present invention also provides a method for preparing the nanoparticles, specifically:

[0016] A dispersion of an organic molecule with ultrasound-activated luminescence properties, an enhancer HBA, and an amphiphilic polymer was mixed, added to water, and sonicated to obtain nanoparticles with ultrasound-activated luminescence properties.

[0017] Specifically, the frequency range of its ultrasound is 30kHz to 3MHz, and the emission wavelength range of organic molecules or nanoparticles is 200 to 2000 nanometers.

[0018] The luminescence time of the nanoparticles ranges from 1 second to 24 hours.

[0019] The luminescence intensity of the nanoparticles is in the range of 10. 2 ~10 10 p / sec.

[0020] The present invention also provides the application of the aforementioned nanoparticles with ultrasound-activated luminescence properties in bioimaging.

[0021] This invention provides a class of organic molecules and nanoparticles with ultrasound-activated luminescence properties. Unlike real-time optical illumination, ultrasound-activated luminescence imaging uses ultrasound as its energy source, resulting in virtually no background noise during imaging. Compared to conventional fluorescence imaging, ultrasound-activated luminescence imaging improves the signal-to-noise ratio, imaging sensitivity, and imaging depth. Furthermore, compared to X-ray activated luminescence, bioluminescence, or Cerenkov luminescence, ultrasound-activated imaging offers several key advantages for in vivo imaging, including radiation-free operation, handheld excitation, ease of operation, safety, and independence from expensive instruments. Therefore, ultrasound-activated luminescence imaging can be effectively used for mapping subcutaneous tumors, peritoneal metastases, and lymph nodes in biological applications.

[0022] Compared with the prior art, the present invention has the following beneficial technical effects:

[0023] (1) Through molecular structure screening, this invention has discovered that organic semiconductor polymer molecules containing porphyrin, cyanine, BODIPY, and thiophene structures, as well as organic semiconductor polymer molecules containing styrene and anthracene derivative structures, possess ultrasound-activated luminescence properties. These ultrasound-activated luminescence nanoparticles can be further obtained by using several types of ultrasound-activated luminescent organic molecules as luminescent agents and amphiphilic polymers as surfactants through ultrasonic treatment.

[0024] (2) The present invention also provides the application of the ultrasound-activated luminescence system in in vivo imaging, which uses organic molecules with ultrasound-activated luminescence properties as luminescent substances to achieve high-performance in vivo imaging applications. Attached Figure Description

[0025] Figure 1 This is a schematic diagram illustrating the construction of the ultrasound-activated luminescent nanoparticles prepared in Example 1.

[0026] Figure 2 The image shows the ultraviolet light spectrum of the nanoparticles prepared in Example 2.

[0027] Figure 3 The image shows the fluorescence of the nanoparticles prepared in Example 2.

[0028] Figure 4 The image shows the ultrasonically activated luminescence of the nanoparticles prepared in Example 2.

[0029] Figure 5 The image shows a TEM image of the TA-NPs prepared in Example 3.

[0030] Figure 6 The images show the ultrasound emission patterns of TA-NPs prepared in Example 3 under different frequencies of ultrasonic excitation.

[0031] Figure 7 The images show the ultrasound emission patterns of TA-NPs obtained in Example 3 under ultrasonic excitation at different times.

[0032] Figure 8 The image shows the sustained ultrasonic emission of TA-NPs obtained in Example 3 after ultrasonic excitation.

[0033] Figure 9 This is a schematic diagram illustrating the construction of PFODBT@HBA-NPs nanoparticles prepared in Example 4.

[0034] Figure 10 The particle size distribution of PFODBT@HBA-NPs obtained in Example 4 is shown.

[0035] Figure 11 The image shows the ultrasonic excitation emission band of PFODBT@HBA-NPs obtained in Example 4.

[0036] Figure 12 The images show the ultrasound emission patterns of PFODBT@HBA-NPs obtained in Example 4 under ultrasonic excitation at different times and the ultrasound emission patterns of PFODBT@HBA-NPs with different concentrations under ultrasonic excitation.

[0037] Figure 13 The images show schematic diagrams (a) and (b) of the ultrasound-excited luminescence imaging of the TA-NP obtained in Example 3.

[0038] Figure 14 The images shown are schematic diagram (a) and imaging diagram (b) of the TA-NP obtained in Example 3, obtained by ultrasound-induced luminescence imaging of subcutaneous tumor.

[0039] Figure 15 The images shown are schematic diagram (a) and imaging diagram (b) of the TA-NP obtained in Example 3, obtained by ultrasound-induced luminescence imaging of peritoneal metastases. Detailed Implementation

[0040] Unless otherwise specified, the experimental methods described in the following examples are conventional methods, and the reagents and materials described are commercially available unless otherwise specified.

[0041] The technical solution of the present invention will be further illustrated below through specific experimental methods.

[0042] Example 1: Synthesis of Nanoparticles with Ultrasonic Activated Luminescence Properties

[0043] This embodiment is based on the synthesis of nanoparticles containing organic molecules with ultrasound-activated luminescence properties. The specific general synthesis method is as follows:

[0044] Nanoparticles based on organic molecules with ultrasound-activated luminescence properties were directly synthesized using a nano-coprecipitation method. First, 1 mL of THF stock solution containing organic molecules (100 μg) and distearate phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG, 2k) (2.5 mg) was prepared. 9 mL of H2O was placed in a serum bottle, and the prepared tetrahydrofuran solution was quickly injected into the water. The mixture was sonicated for 8 minutes. After removing THF by rotary evaporation, the mixture was centrifuged (4000 rpm, 5 minutes), washed with deionized water, concentrated, and stored in the dark.

[0045] The construction of ultrasonically activated luminescent nanoparticles in this embodiment is as follows: Figure 1 As shown.

[0046] Example 2: Property Testing of Nanoparticles with Ultrasonic Activated Luminescence Properties

[0047] This embodiment is based on the property study of nanoparticles with ultrasound-activated luminescence properties. The specific research content is as follows:

[0048] (1) Study on the ultraviolet properties of nanoparticles with ultrasound-activated luminescence: The obtained nanoparticles were diluted to a certain extent, and then the ultraviolet absorption spectrum of the molecules was measured by an ultraviolet instrument. The results are as follows: Figure 2 As shown.

[0049] (2) Study on the fluorescence properties of nanoparticles with ultrasound-activated luminescence: The obtained nanoparticles were diluted to a certain extent, and then the fluorescence emission spectrum of the molecules was measured by a fluorescence spectrometer. The results are as follows: Figure 3 As shown.

[0050] (3) Study on ultrasonic-activated luminescence of nanoparticles with ultrasonic-activated luminescence properties: We used a small animal imaging instrument to collect ultrasonic luminescence signals of nanoparticles.

[0051] from Figure 4 It can be seen that, at the same mass concentration, the ultrasonic-activated luminescence ability of TA-based nanoparticles (TA-NPs) is the strongest.

[0052] Example 3: Synthesis and Property Verification Based on TA-NPs

[0053] This embodiment is based on the study of the properties of TA-NPs, and the specific synthesis steps are as described in Example 1.

[0054] Figure 5 The TEM image of the TA-NPs obtained in Example 3 is shown below. Figure 5 It can be seen that the synthesized particles have a diameter of approximately 30 nanometers.

[0055] (1) Study on the wavelength of ultrasound-activated emission of TA-NPs: The obtained TA-NPs were ultrasound-activated for the same time using ultrasound generators of different frequencies, and then the ultrasound-activated emission was collected by Edinburgh fluorescence spectrometer.

[0056] from Figure 6 It can be seen that the ultrasound-activated luminescence of TA-NPs is mainly concentrated in the 600-650 nm range, which is similar to its fluorescence emission spectrum.

[0057] (2) Study on ultrasound-activated luminescence properties of TA-NPs: The obtained TA-NPs were irradiated with ultrasound for different times, and then the luminescence images of TA-NPs after ultrasound for different times were tested by a small animal imaging instrument.

[0058] from Figure 7 It can be seen that the intensity of ultrasound-activated luminescence of TA-NPs increases with the extension of ultrasound time.

[0059] (3) Study on ultrasound-activated luminescence time of TA-NPs: After ultrasounding the obtained TA-NPs for 15 seconds, afterglow luminescence images were collected at different time points after ultrasounding stopped using a small animal imaging device.

[0060] from Figure 8 It can be seen that the ultrasound-activated luminescence signal of TA-NPs can still be collected 10 minutes after the ultrasound excitation stops.

[0061] Example 4: Synthesis and Property Verification of PFODBT@HBA-NPs

[0062] This embodiment is based on the synthesis and property study of PFODBT@HBA-NPs. The specific synthesis method is as follows:

[0063] (1) Preparation of PFODBT@HBA-NPs nanoparticles:

[0064] Nanoparticles based on PFODBT@HBA-NPs were directly synthesized using a nano-coprecipitation method: First, 1 mL of a THF stock solution containing PFODBT (100 μg), HBA (100 μg), distearate phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG, 2k) (2.5 mg) was prepared. 9 mL of H2O was placed in a serum bottle, and the prepared tetrahydrofuran solution was quickly injected into the water. The solution was sonicated for 8 minutes. After removing THF by rotary evaporation, the solution was centrifuged (4000 rpm, 5 minutes), washed with deionized water, concentrated, and stored in the dark.

[0065] The construction of ultrasonically activated luminescent nanoparticles in this embodiment is as follows: Figure 9 As shown.

[0066] Figure 10 The images shown are TEM and DLS images of PFODBT@HBA-NPs obtained in Example 4. Figure 10 It can be seen that the synthesized particles have a diameter of approximately 30 nanometers.

[0067] (2) Study on ultrasound-activated emission bands of PFODBT@HBA-NPs: The obtained PFODBT@HBA-NPs were ultrasound-activated for a period of time, and then ultrasound-activated emission images were collected in different bands (510-570 nm, 570-650 nm, 690-700 nm, 800-820 nm) using a small animal imaging instrument.

[0068] from Figure 11 It can be seen that the ultrasound-activated luminescence of PFODBT@HBA-NPs is mainly concentrated in the 510-570 nm range.

[0069] (3) Study on the ultrasound-activated luminescence properties of PFODBT@HBA-NPs: The obtained PFODBT@HBA-NPs were irradiated with ultrasound for different times, and then the luminescence images of PFODBT@HBA-NPs after ultrasound for different times were tested using a small animal imaging device. In addition, PFODBT@HBA-NPs of different concentrations were ultrasound-activated for 15 seconds, and their light images were tested using a small animal imaging device.

[0070] from Figure 12 It can be seen that the intensity of ultrasound-activated luminescence of PFODBT@HBA-NPs increases with the extension of ultrasound time. The intensity of ultrasound-activated luminescence of PFODBT@HBA-NPs also exhibits concentration dependence; the higher the concentration, the stronger the luminescence intensity.

[0071] Example 5: Ultrasound-induced luminescence imaging of lymph nodes based on TA-NPs

[0072] For ultrasound activation imaging of mouse lymph nodes, mice were anesthetized with 2% isoflurane oxygen, and TA-NPs were injected into the hind paws. Ultrasound coupling gel was applied to the lymph node sites, and the ultrasound transducer was placed firmly against the ultrasound coupling gel at 30 kHz, 4.5 W / cm². 2 Fifteen seconds later, the mice were immediately imaged using a small animal imaging device to image the lymph node region.

[0073] Figure 13 The images shown are schematic diagrams (a) and (b) of lymph node ultrasound-activated chemiluminescence imaging after TA-NPs were injected into the hind paw in Example 5.

[0074] Example 6: Ultrasound-induced emission imaging of tumors based on TA-NPs

[0075] For ultrasound activation imaging of subcutaneous tumors, mice were anesthetized with 2% isoflurane oxygen, and TA-NPs were injected intratumorally. Ultrasound coupling gel was applied between the ultrasound transducer and the tumor. Ultrasound was then administered at 30 kHz, 4.5 W / cm². 2 15 seconds later, the mice were immediately imaged using a small animal imaging device to image the subcutaneous tumor area.

[0076] Figure 14 The images shown are schematic diagrams (a) and (b) of ultrasound-activated chemiluminescence imaging of subcutaneous tumors after intratumoral injection of TA-NPs obtained in Example 6.

[0077] For ultrasound activation imaging of peritoneal metastases, mice were anesthetized with 2% isoflurane oxygen, and TA-NPs were injected via the tail vein. Ultrasound coupling gel was applied to the lower abdomen of the mice, and the ultrasound transducer was placed firmly against the ultrasound coupling gel on the lower abdomen of the mice, using ultrasound (30kHz, 4.5W / cm²). 2 Fifteen seconds later, the mice were immediately imaged using a small animal imaging system to visualize the metastatic tumors in the lower abdominal region.

[0078] Figure 15 The images shown are schematic diagrams (a) and (b) of ultrasound-activated chemiluminescence imaging of subcutaneous tumors after intratumoral injection of TA-NPs obtained in Example 6.

[0079] The above description is only a preferred embodiment of the present invention. All equivalent changes and modifications made within the scope of the claims of the present invention should be included in the scope of the present invention.

Claims

1. Nanoparticles having ultrasound-activated luminescence properties, characterized in that, The nanoparticles are made of organic molecules, a luminescence enhancer HBA; The structure of the organic molecules is as follows: The HBA has the following structural formula: HBA The nanoparticles are uniform spherical structures; The size of the nanoparticles is 1-1000 nanometers, the ultrasonic frequency range for exciting the nanoparticles is 30 kHz-3 MHz, the emission wavelength range of the organic molecules or the nanoparticles is 200-2000 nanometers, and the luminescence time is 1 second to 24 hours; Its luminous intensity ranges from 10 2 ~10 10 p / sec.

2. The nanoparticles having the property of ultrasound-activated luminescence according to claim 1, characterized in that, The nanoparticles are prepared by a self-assembly method with organic molecules as a core and amphiphilic polymers as a surface modification layer.

3. Use of the nanoparticles with the ultrasonic-activated luminescence property according to claim 1 or 2 in the preparation of a biological imaging reagent.

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