Nanoparticles with ultrasound-activated luminescence properties, methods of preparation and applications
By preparing nanoparticles that are self-assembled from chlorophyll and porphyrin molecules, the problems of limited types of luminescent reagents and low energy conversion efficiency in ultrasound-induced luminescence technology have been solved, enabling high-performance in vivo imaging and localization of deep tissue lesions. This is particularly suitable for precise imaging of subcutaneous tumors and peritoneal metastases.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- HANGZHOU INSTITUTE OF MEDICAL SCIENCES CHINESE ACADEMY OF SCIENCES
- Filing Date
- 2026-04-24
- Publication Date
- 2026-07-14
AI Technical Summary
The limited variety of luminescent reagents and unsatisfactory energy conversion efficiency in existing ultrasound-induced luminescence technology restrict its application in deep tumor diagnosis and biomarker monitoring.
Nanoparticles with ultrasound-activated luminescence properties were prepared by using nanoparticles formed by the self-assembly of chlorophyll molecules and porphyrin molecules, with an amphiphilic polymer as a surface modification layer and ultrasonic treatment, thereby enhancing the luminescence intensity and tissue penetration ability.
It achieves highly sensitive in vivo imaging, improves imaging depth and signal-to-noise ratio, and features no ionizing radiation, safe and convenient operation. It is suitable for precise localization of subcutaneous tumor boundaries and peritoneal metastases.
Smart Images
Figure CN122376793A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of luminescent materials, specifically relating to a nanoparticle with ultrasonically activated luminescence properties, its preparation method, and its application. Background Technology
[0002] Optical imaging plays a crucial role in disease diagnosis, detection, prediction, and monitoring due to its advantages such as high sensitivity, high spatiotemporal resolution, and ease of operation. Traditional fluorescence imaging requires real-time photoexcitation, which is limited by tissue autofluorescence, resulting in a reduced signal-to-background ratio, limited imaging depth (<1 cm), and decreased sensitivity. This has led to the development of autofluorescence-free imaging modalities. Currently, these strategies mainly include five modalities: bioluminescence, chemiluminescence, afterglow luminescence, Cherenkov radiation, and X-ray induced luminescence. Although these methods eliminate excitation-related background signals, each has its limitations: bioluminescence / chemiluminescence intensity is affected by the microenvironment; the application of afterglow probes in deep lesions is limited by insufficient penetration depth due to in-situ pre-excitation of the light source; Cherenkov imaging requires high-dose radioactive tracers, and X-ray induced luminescence relies on cumulative radiation dose to obtain sufficient contrast, posing a challenge to safe longitudinal monitoring. Ultrasound-induced luminescence, as an emerging modality, utilizes mechanical ultrasound to excite molecules to produce optical emission. Ultrasound combines clinical maturity and safety, possesses deep tissue penetration capabilities, and can achieve optical reading of deep targets without optical pre-excitation or biochemical mediation. Meanwhile, the physical decoupling characteristics of ultrasonic excitation and optical output further reduce background interference, achieving a high signal-to-background ratio and enhanced detection sensitivity, making it suitable for sensitive imaging of anatomical and optically inaccessible tissues.
[0003] Ultrasound-induced luminescence (UIL) is a novel optical technique that converts mechanical ultrasound energy into molecular luminescence. Traditional molecular optical imaging relies primarily on electromagnetic wave energy absorption-induced electronic transitions and radiative relaxation, while ultrasound, as mechanical energy (frequency >20 kHz), cannot directly excite molecules through ground-state-excited state transitions. UIL overcomes this limitation through an energy conversion mechanism: ultrasound mechanical energy is first converted into chemical energy, which then drives electrons in the luminescent molecules to an excited state, generating detectable photons upon returning to the ground state. This technique offers advantages such as deep tissue penetration, independence from the microenvironment (insensitive to enzyme conditions and substrate distribution), and operational safety, making it suitable for deep tumor diagnosis and monitoring of specific biomarkers. However, the limited variety of luminescent reagents and the less-than-ideal energy conversion efficiency currently hinder its wider biological and clinical applications. Summary of the Invention
[0004] To address the shortcomings of existing technologies, the present invention aims to provide nanoparticles with ultrasound-activated luminescence properties, their preparation method, and their applications, in order 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 a nanoparticle with ultrasound-activated luminescence properties, which is self-assembled from chlorophyll molecules, porphyrin molecules and amphiphilic polymers;
[0007] The structural formula of the chlorophyll molecule includes:
[0008] The structural formulas of the porphyrin molecules include: .
[0009] In a preferred embodiment, the nanoparticles have a uniform spherical structure, with chlorophyll molecules and porphyrin molecules having ultrasound-activated luminescence properties as the core, and an amphiphilic polymer as the surface modification layer.
[0010] Furthermore, the size of the nanoparticles is 1 to 1000 nanometers.
[0011] In a preferred embodiment, the amphiphilic polymer is any one or a combination of poly(2-(methacryloyloxy)ethyldimethyl(3-sulfopropyl)ammonium hydroxide), poly(styrene-maleic anhydride), and sodium dodecylbenzenesulfonate.
[0012] Furthermore, the amphiphilic polymer is poly(styrene-maleic anhydride).
[0013] This invention also provides a method for preparing the aforementioned nanoparticles with ultrasound-activated luminescence properties, specifically:
[0014] A dispersion of chlorophyll molecules, porphyrin molecules, and amphiphilic polymers is mixed, added to water, and ultrasonically treated to obtain the nanoparticles with ultrasonically activated luminescence properties, i.e., the luminescent system.
[0015] Specifically, the wavelength range of the nanoparticles is 200~1000 nanometers.
[0016] The luminescent system described above can emit light continuously after ultrasonic excitation.
[0017] Specifically, its ultrasonic frequency range is 30kHz ~ 3MHz.
[0018] Specifically, its ultrasound-induced luminescence time ranges from 1 second to 24 hours.
[0019] Specifically, its ultrasound-induced luminescence intensity ranges from 10. 2 ~10 10 p / sec.
[0020] Specifically, the luminescent system also includes the reinforcing agent SO;
[0021] The reinforcing agent SO itself does not emit light, and the reinforcing agent SO is N,N-dimethyl-4-(2-phenyl-5,6-dihydro-1,4-oxothionadien-3-yl)aniline.
[0022] The present invention also provides the application of the aforementioned nanoparticles with ultrasound-activated luminescence properties, in the preparation of in vivo imaging reagents.
[0023] This invention provides nanoparticles with ultrasound-activated luminescence properties. Ultrasound-activated luminescence imaging uses ultrasound instead of traditional light sources as the excitation energy source, fundamentally solving the bottleneck of low signal-to-noise ratio in traditional optical imaging due to its unique "no background noise" mechanism. Compared to conventional fluorescence imaging, this technology significantly improves imaging sensitivity and tissue penetration depth. Compared to other deep tissue imaging modalities such as X-ray activated luminescence, bioluminescence, and Cerenkov luminescence, it possesses key advantages such as no ionizing radiation, safe and convenient operation, handheld excitation, and no reliance on expensive equipment. With these characteristics, ultrasound-activated luminescence imaging demonstrates unique imaging value in in vivo applications such as subcutaneous tumor delineation and precise localization of peritoneal metastases.
[0024] Compared with the prior art, the present invention has the following beneficial technical effects:
[0025] (1) This invention uses chlorophyll molecules that can be activated by ultrasound to emit light as the light source through molecular structure screening, and uses amphiphilic polymers as surfactants to obtain nanoparticles that can be activated by ultrasound through ultrasonic treatment.
[0026] (2) The present invention also provides the application of the ultrasound-activated luminescence system in the imaging of organisms. The system uses chlorophyll molecules with ultrasound-activated luminescence properties as luminescent substances, and can construct an ultrasound-activated luminescence system to realize high-performance in vivo imaging applications for locating deep tissue lesions and sensing disease biomarkers. Attached Figure Description
[0027] Figure 1 This is a schematic diagram of the preparation of ultrasonically activated luminescent nanoparticles prepared in Example 1.
[0028] Figure 2 The image shows the luminescence of the nanoparticles prepared in Example 2.
[0029] Figure 3 TEM image and particle size distribution of CB NPs prepared in Example 3.
[0030] Figure 4 The image shows the ultrasonic excitation emission band diagram of the CB NPs obtained in Example 3.
[0031] Figure 5The images show the ultrasound emission patterns of CB NPs obtained in Example 3 under ultrasonic excitation at different times.
[0032] Figure 6 The image shows the ultrasonic emission patterns of CB NPs obtained in Example 3 under different power ultrasonic excitation.
[0033] Figure 7 The image shows the ultrasound luminescence of CB NPs obtained in Example 3 under different thicknesses of chicken tissue.
[0034] Figure 8 The images shown are schematic diagrams (a) and (b) of the peritoneal cavity obtained by ultrasound-induced emission imaging of CB@SO-NPs obtained in Example 4.
[0035] Figure 9 The images shown are schematic diagram (a) and imaging diagram (b) of subcutaneous tumors obtained by ultrasound-induced luminescence imaging of CB@SO-NPs obtained in Example 5.
[0036] Figure 10 The images shown are schematic diagrams (a) and (b) of ultrasound-induced luminescence imaging of CB@SO-NPs obtained in Example 5 for in situ pancreatic cancer. Detailed Implementation
[0037] 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.
[0038] The technical solution of the present invention will be further illustrated below through specific experimental methods.
[0039] Example 1: Synthesis of Nanoparticles with Ultrasonic Activated Luminescence Properties
[0040] This embodiment is based on the synthesis of nanoparticles containing chlorophyll molecules and porphyrin molecules with ultrasound-activated luminescence properties. The specific general synthesis method is as follows:
[0041] Nanoparticles containing chlorophyll and porphyrin molecules with ultrasound-activated luminescence properties were directly synthesized using a nano-coprecipitation method. First, 1 mL of THF stock solution containing chlorophyll and porphyrin molecules (50 μg) and poly(styrene-maleic anhydride) (PSMA) (6 mg) was prepared. 9 mL of H2O was placed in a serum bottle, and the prepared tetrahydrofuran solution was quickly injected into the water. After 1 minute, 1 mL of NaHCO3 (10 mg / mL) was injected, and the mixture was sonicated for 5 minutes. After removing THF by rotary evaporation, the mixture was centrifuged (3000 rpm, 5 minutes), washed with deionized water, concentrated, and stored in the dark.
[0042] The construction of ultrasonically activated luminescent nanoparticles in this embodiment is as follows: Figure 1As shown.
[0043] Example 2: Property Testing of Nanoparticles with Ultrasonic Activated Emissivity
[0044] This embodiment is based on the property study of nanoparticles with ultrasound-activated luminescence properties. The specific research content is as follows:
[0045] Study on ultrasound-activated luminescence of nanoparticles with ultrasound-activated luminescence properties: We used a small animal imaging system to collect ultrasound luminescence and fluorescence signals of the nanoparticles.
[0046] from Figure 2 It can be seen that, at the same mass concentration, the ultrasonic-activated luminescence ability of CB molecule-based nanoparticles (CB NPs) is the strongest.
[0047] Example 3: Synthesis and Property Verification of CB NPs
[0048] This embodiment is based on the synthesis and property study of CB NPs, and the specific synthesis steps are as in Example 1.
[0049] Figure 3 The TEM and DLS images of CB NPs obtained in Example 3 are shown below. Figure 9 It can be seen that the synthesized particles have a diameter of approximately 30 nanometers.
[0050] (1) Study on ultrasound-activated emission bands of CB NPs: The obtained CB NPs were ultrasound-activated for a period of time, and then ultrasound-activated emission images were collected by small animal imaging device in different bands (570~620 nm, 620~670 nm, 670~710 nm, 710~790 nm).
[0051] from Figure 4 It can be seen that the ultrasound-activated luminescence of CB NPs is mainly concentrated at 670 nm.
[0052] (2) Study on the ultrasound-activated luminescence properties of CB NPs: The obtained CB NPs were ultrasound-excited for different times, and then the luminescence images of CB NPs after ultrasound for different times were tested using a small animal imaging device. In addition, CB NPs of different concentrations were ultrasound-excited for 60 seconds, and their light images were tested using a small animal imaging device.
[0053] from Figure 5 It can be seen that the intensity of ultrasound-activated luminescence of CB NPs increases with the extension of ultrasound time. The intensity of ultrasound-activated luminescence of CB NPs also exhibits concentration dependence; the higher the concentration, the stronger the luminescence intensity.
[0054] (4) Study on ultrasound-activated luminescence properties of CB NPs: The obtained CB NPs were excited for the same time with different ultrasound powers, and then the luminescence images of CB NPs after ultrasound with different powers were tested by a small animal imaging instrument.
[0055] from Figure 6 It can be seen that the intensity of ultrasound-activated emission of CB NPs increases with increasing ultrasound power.
[0056] (5) Tissue penetration study of ultrasound-activated luminescence of CB NPs: After ultrasounding muscle tissue of different thicknesses with the obtained CB NPs for 120 seconds, the corresponding thickness of tissue was covered after the ultrasound stopped and the luminescence images were collected by a small animal imaging device.
[0057] from Figure 7 It can be seen that the ultrasound-activated luminescence of CB NPs can penetrate 4 cm of chicken tissue, and the signal-to-background ratio is higher than that of fluorescence.
[0058] Example 4: Ultrasound-induced luminescence imaging of the peritoneal cavity based on CB@SO-NPs
[0059] For intraperitoneal ultrasound activation imaging in mice, mice were anesthetized with 2% isoflurane oxygen, and CB@SO-NPs were injected into the peritoneal cavity. Ultrasound coupling gel was applied to the abdominal region of the mice, and the ultrasound transducer was placed firmly against the lymph node site on the ultrasound coupling gel. Ultrasound (1 MHz, 3 W / cm²) was then applied. 2 60 seconds later, the mice were immediately imaged using a small animal imaging system to visualize the peritoneal cavity region.
[0060] Figure 8 The images shown are schematic diagrams (a) and (b) of abdominal ultrasound-activated luminescence imaging of intraperitoneal injection of CB@SO-NPs obtained in Example 4.
[0061] Example 5: Ultrasound-induced emission imaging of tumors based on CB@SO-NPs
[0062] For ultrasound activation imaging of subcutaneous tumors, mice were anesthetized with 2% isoflurane oxygen, and CB@SO-NPs were injected via the tail vein. Ultrasound coupling gel was applied between the ultrasound transducer and the tumor. Ultrasound was then applied (1 MHz, 3 W / cm²). 2 60 seconds later, the mice were immediately imaged using a small animal imaging system to image the subcutaneous tumor area.
[0063] Figure 9 The images shown are schematic diagrams (a) and (b) of ultrasound-activated chemiluminescence imaging of subcutaneous tumors after tail vein injection of CB@SO-NPs obtained in Example 5.
[0064] For ultrasound activation imaging of in situ pancreatic cancer, mice were anesthetized with 2% isoflurane oxygen, and CB@SO-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. Ultrasound was applied at 1 MHz, 3 W / cm². 2 Sixty seconds later, the mice were immediately imaged using a small animal imaging system to visualize the metastatic tumors in the lower abdominal region.
[0065] Figure 10 The images shown are schematic diagrams (a) and (b) of ultrasound-activated chemiluminescence imaging of in situ pancreatic cancer after tail vein injection of CB@SO-NPs obtained in Example 5.
[0066] The above description is only a part of the embodiments of the present invention and does not limit the scope of protection of the present invention. Any equivalent device or equivalent process transformation made based on the content of the present invention specification and drawings, or direct or indirect application in other related technical fields, are similarly included within the patent protection scope of the present invention.
Claims
1. A nanoparticle with ultrasound-activated luminescence properties, characterized in that, These nanoparticles are self-assembled from chlorophyll molecules, porphyrin molecules, and amphiphilic polymers. The structural formula of the chlorophyll molecule includes: The structural formulas of the porphyrin molecules include: .
2. The nanoparticle with ultrasound-activated luminescence properties according to claim 1, characterized in that, The nanoparticles have a uniform spherical structure, with chlorophyll molecules and porphyrin molecules having ultrasound-activated luminescence properties as the core, and an amphiphilic polymer as the surface modification layer.
3. The nanoparticle with ultrasound-activated luminescence properties according to claim 2, characterized in that, The size of the nanoparticles is 1 to 1000 nanometers.
4. The nanoparticle with ultrasound-activated luminescence properties according to claim 1, characterized in that, The amphiphilic polymer is any one or a combination of poly(2-(methacryloyloxy)ethyldimethyl(3-sulfopropyl)ammonium hydroxide), poly(styrene-maleic anhydride), and sodium dodecylbenzenesulfonate.
5. The nanoparticle with ultrasound-activated luminescence properties according to claim 4, characterized in that, The amphiphilic polymer is poly(styrene-maleic anhydride).
6. The method for preparing nanoparticles with ultrasound-activated luminescence properties according to any one of claims 1 to 5, characterized in that, Specifically: A dispersion of chlorophyll molecules, porphyrin molecules, and amphiphilic polymers is mixed, added to water, and ultrasonically treated to obtain the nanoparticles with ultrasonically activated luminescence properties, i.e., the luminescent system.
7. The method for preparing nanoparticles with ultrasound-activated luminescence properties according to claim 6, characterized in that, The wavelength range of the nanoparticles is 200~1000 nanometers.
8. The method for preparing nanoparticles with ultrasound-activated luminescence properties according to claim 6, characterized in that, The luminescent system described above can emit light continuously after ultrasonic excitation; Its ultrasonic frequency range is 30kHz ~ 3MHz; Its ultrasound-induced luminescence time ranges from 1 second to 24 hours; Its ultrasound-induced luminescence intensity ranges from 10 2 ~10 10 p / sec.
9. The method for preparing nanoparticles with ultrasound-activated luminescence properties according to claim 6, characterized in that, The luminescent system also includes an enhancer SO; The reinforcing agent SO itself does not emit light, and the reinforcing agent SO is N,N-dimethyl-4-(2-phenyl-5,6-dihydro-1,4-oxothionadien-3-yl)aniline.
10. The application of the nanoparticles with ultrasound-activated luminescence properties according to any one of claims 1 to 9, characterized in that, It was used to prepare in vivo imaging reagents.