Ultrasonic activated rhenium cyanine dye complex nano preparation and application thereof

Abstract

The invention provides an ultrasonic activated rhenium cyanine dye complex nano preparation, which is characterized in that heptamethine cyanine dye (IR-780) and rhenium (I) tricarbonyl center are coupled through a covalent bond to construct a novel complex Re-Cy7. According to the molecular design, ultrasonic synchronous triggering dual effects are achieved by expanding a pi-conjugated system, oxygen-independent-form. OH is efficiently generated through an I-type mechanism, and a therapeutic dose of CO is accurately released. Furthermore, a nano preparation (Re-Cy7NPs) is constructed through self-assembly of Pluronic F-127, so that the water solubility and the targeted accumulation capacity of an infected part are remarkably improved. The technology breaks through the existing limitation at one stroke: the restriction of an anoxic environment on SDT is overcome, the constraint of light penetration depth on gas release is relieved, efficient synergy of. OH and CO is realized on a single molecule level, and a brand new solution is provided for radically treating MDR infection.

Description

Technical Field

[0001] This invention relates to the field of materials for sonodynamic therapy and gas therapy, and specifically to an ultrasound-activated rhenium cyanine dye complex nanoparticle and its application. Background Technology

[0002] Global efforts to control bacterial infections are facing severe challenges, with high morbidity and mortality rates continuously increasing the burden on public health. The long-term overuse of traditional antibiotics has led to the widespread spread of multidrug-resistant (MDR) bacteria, significantly diminishing the efficacy of existing antimicrobial drugs and making bacterial infections a major health threat in the 21st century. In particular, the rapid evolution of MDR strains and the increasing complexity of their resistance mechanisms necessitate a breakthrough in traditional treatment paradigms and the development of novel antimicrobial strategies.

[0003] Sonodynamic therapy (SDT), as a non-invasive treatment, uses ultrasound (US) to activate a sonosensitive agent to generate reactive oxygen species (ROS) that kill pathogens. It has significant advantages such as deep tissue penetration, strong spatiotemporal controllability, and low susceptibility to inducing drug resistance. However, current SDT technology heavily relies on type II sonosensitive agents (such as porphyrin derivatives), which generate singlet oxygen through energy transfer (…). 1 The mechanism of O2 is highly dependent on an oxygen-rich environment. However, the hypoxic microenvironment commonly found in bacterial infection lesions greatly limits the ability of bacteria to thrive. 1 The reduced efficiency of O2 generation leads to a sharp drop in efficacy, becoming a core bottleneck restricting the clinical translation of SDT. Meanwhile, organic acoustic sensitizers (such as cyanine dyes) are prone to degradation under ultrasound / light irradiation, but as acoustic sensitizers, they need to maintain structural integrity to continuously generate reactive oxygen species.

[0004] Gas therapy utilizes endogenous gaseous signaling molecules (such as CO and NO) to regulate biological processes, providing a new approach to anti-infective treatment. CO has been proven to have multiple therapeutic values: it can directly disrupt bacterial biofilm structures, inhibit electron transport in the pathogen respiratory chain, enhance the phagocytic activity of macrophages against drug-resistant bacteria, and promote angiogenesis and epithelial regeneration in infected tissues. More importantly, CO is less likely to induce bacterial resistance. However, targeted delivery and controlled release of CO remain unsolved challenges: systemic release of free CO can easily cause tissue toxicity (such as hypoxia due to hemoglobin binding); existing CO-releasing molecules (CORMs) mostly rely on photoactivation (such as rhenium / manganese carbonyl complexes), but the tissue penetration depth of ultraviolet-visible light is insufficient, making it unable to effectively target deep infection foci; and high-energy light irradiation may damage normal tissues, posing safety risks.

[0005] Studies have shown that the synergistic effect of ROS and gas molecules can significantly improve antibacterial efficiency. For example, ·OH can cleave the biofilm matrix and promote CO permeation, while CO can inhibit the bacterial antioxidant system and amplify the killing effect of ROS. However, the deep integration of SDT and CO therapy is still in the exploratory stage. Current reports mostly use physical mixing systems or simple nanocarriers loaded with CO release agents. Their synergistic effect is only manifested in spatial coexistence or ROS indirectly promoting CO release, lacking a bifunctional integrated system designed at the molecular level. Such systems cannot achieve the spatiotemporal coupling of ultrasonic synchronous triggering of the generation of potent bactericidal factors (such as ·OH) and precise CO release, limiting the depth of the synergistic effect. Moreover, existing materials have poor controllability, especially lacking the ability to efficiently utilize oxygen-independent potent bactericidal ROS (such as ·OH). Summary of the Invention

[0006] The purpose of this invention is to overcome at least one deficiency of the prior art and to provide an ultrasonically activated rhenium cyanine dye complex nanoformation and its application.

[0007] The technical solution adopted in this invention is:

[0008] In a first aspect, the present invention provides an ultrasonically activated rhenium cyanine dye complex nanoparticle, wherein the preparation method of the rhenium cyanine dye complex nanoparticle includes the following steps:

[0009] 1) Under strictly anhydrous conditions, the 1,10-o-phenanthroline derivative ligand was deprotonated using sodium hydride, and then the deprotonated product was reacted with IR-780 under an inert gas atmosphere to obtain the intermediate IR780-phen.

[0010] 2) The complex Re-Cy7 was synthesized by reacting Re(CO)5Cl with IR780-phen under an anhydrous and inert gas atmosphere;

[0011] 3) The complex Re-Cy7 was added to a solution composed of Pluronic F-127 (F-127) and phosphate buffer (PBS) to finally obtain the rhenium cyanine dye complex nanoparticles Re-Cy7 NPs.

[0012] In some embodiments, the molar ratio of sodium hydride to 1,10-phenanthroline derivative ligand in step 1) is 0.9-1.1.

[0013] In some embodiments, the molar ratio of Re(CO)5Cl and IR780-phen in step 2) is 0.9-1.1.

[0014] In some embodiments, the reaction temperature of step 2) is 110±2℃.

[0015] In some embodiments, the reaction time of step 2) is 4-5 hours.

[0016] In some embodiments, the molar ratio of the deprotonation product to IR-780 in step 1) is 1.9-2.1.

[0017] The second aspect is the application of the rhenium cyanine dye complex nanoformulation described in the first aspect of the present invention in the preparation of formulations for treating bacterial infections and promoting wound healing.

[0018] In some embodiments, the formulation is a sonodynamic therapy formulation.

[0019] In some embodiments, the formulation is a gas therapy formulation.

[0020] In some embodiments, the formulation is a therapeutic formulation for sonodynamic therapy combined with gas therapy.

[0021] The beneficial effects of this invention are:

[0022] To address the aforementioned bottlenecks, this invention proposes an innovative intramolecular bifunctional integrated solution: a novel complex, Re-Cy7, is constructed by covalently coupling heptamethine dye (IR-780) with the rhenium (I) tricarbonyl center. This molecular design utilizes an extended π-conjugated system to achieve a dual effect triggered by ultrasound—efficiently generating oxygen-independent ·OH via a type I mechanism and precisely releasing a therapeutic dose of CO. Furthermore, nano-formulations (Re-Cy7 NPs) are constructed through the self-assembly of Pluronic F-127, significantly enhancing water solubility and targeted accumulation at infection sites. This technology overcomes existing limitations: it overcomes the constraints of hypoxic environments on SDT, removes the limitations of light penetration depth on gas release, and achieves highly efficient synergy between ·OH and CO at the single-molecule level, providing a novel solution for the radical treatment of MDR infections. Attached Figure Description

[0023] Figure 1 Characterization diagrams of Re-Cy7 NPs are shown, including (a) TEM images of Re-Cy7 NPs; (b) hydrodynamic size distribution of Re-Cy7 NPs; (c) UV-Vis absorption spectra of Re-Cy7 and Re-Cy7 NPs in PBS; (d) IR spectral analysis of Re-Cy7 molecules, Re-Cy7 NPs, and Pluronic F-127 (F-127); (e) Zeta potential (mV) of Re-Cy7 NPs; and (f) hydrodynamic diameter changes of Re-Cy7 NPs over 7 days in PBS, DMEM, and deionized (DI) water.

[0024] Figure 2ESR spectrum of ·OH generated by complex Re-Cy7 (100 μM) under sonication (using DMPO (25 mM) as spin trapping agent).

[0025] Figure 3 Figure 1.1: Methylene blue (MB, ·OH probe) in aqueous solution with and without ultrasonic irradiation (1.0 MHz, 1.4 W / cm²) in the presence of Re-Cy7 (Figure 1a) and IR-780 (Figure 1b) (10 μM). 2 The change of the UV-Vis absorption spectrum with irradiation time at 50% duty cycle.

[0026] Figure 4 In the presence of Re-Cy7 (10 μM), CAY10732 (10 μM, excitation wavelength: 488 nm) under ultrasonic irradiation (1.0 MHz, 1.4 W / cm²) 2 The fluorescence intensity changes over time at 50% duty cycle.

[0027] Figure 5 To detect changes in the UV-Vis absorption spectrum of hemoglobin (Hb, 4 μM), ultrasound irradiation (1.0 MHz, 1.4 W / cm²) was performed. 2 CO released over time by Re-Cy7 (1 μM) at 50% duty cycle.

[0028] Figure 6 The in vitro antibacterial activity of Re-Cy7 NPs against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus (MRSA) under ultrasonic conditions is shown in Figures (a, c, e, g, i). Inhibition rates of different concentrations of Re-Cy7 NPs against Staphylococcus aureus (a), Bacillus subtilis (c), Escherichia coli (e), Pseudomonas aeruginosa (g), and MRSA (i) are also shown. (b, d, f, h, j) LB agar plates containing bacteria treated with 16 μM Re-Cy7 NPs are shown: Staphylococcus aureus (b), Bacillus subtilis (d), Escherichia coli (f), Pseudomonas aeruginosa (h), and MRSA (j).

[0029] Figure 7 The image shows the in vivo antibacterial activity and wound healing effect of Re-Cy7 NPs.

[0030] Figure 8 This is a graph showing the change in mouse body weight after drug treatment. Detailed Implementation

[0031] The present invention will be further described below with reference to specific embodiments, but the present invention is not limited thereto.

[0032] Example

[0033] The specific method for synthesizing Re-Cy7 NPs nanoparticles in this embodiment is as follows:

[0034] 1. Synthesis of ligand IR780-phen

[0035] The synthesis equation for the ligand IR780-phen is shown below. Under strictly anhydrous conditions, Phen-OH (620 mg, 1.6 mmol) and an equimolar amount of NaH (64 mg, 1.6 mmol) were dissolved separately in dehydrated N,N-dimethylformamide (DMF, 5 mL), and the entire process was carried out under N2 protection. The two solutions were combined in a dry flask and stirred for 10 minutes to complete deprotonation. A dehydrated DMF solution (1 mL) of IR-780 (533 mg, 0.8 mmol) was added dropwise to the system, and the mixture was stirred continuously at room temperature for 24 hours in the dark. The crude product was extracted with dichloromethane (20 mL × 3), and the organic phases were combined and concentrated under reduced pressure. Finally, it was purified by rapid silica gel column chromatography (eluent: dichloromethane / methanol = 30:1, v / v) to obtain a dark green solid IR780-phen in 45% yield.

[0036]

[0037] Characterization data of ligand IR780-phen: 1 H NMR(400MHz, DMSO-d6)δ13.71(s,1H),9.04(d,J=4.2Hz,2H),8.98–8.84(m,2H),8.36( d,J=8.8Hz,2H),7.85(ddd,J=12.5,8.0,3.8Hz,4H),7.56–7.32(m,8H),7.17(t,J=7.2 Hz,2H),6.26(d,J=14.3Hz,2H),4.14(t,J=7.0Hz,4H),2.78(d,J=5.4Hz,4H),2.08–1. 96(m,2H),1.81–1.67(m,4H),1.33(s,12H),0.94(t,J=7.4Hz,6H).ESI-MS(CH3OH):m / z 815.30[M] + .

[0038] 2. Synthesis of the complex Re-Cy7

[0039] The synthesis reaction equation for the complex Re-Cy7 is shown below. Re(CO)₅Cl (1.4 mmol) and IR780-phen (1.5 mmol) were dissolved in 70 mL of molecular sieve-dehydrated toluene and refluxed at 110 ± 2 °C for 4 h under N₂ protection (precise temperature control in an oil bath). After the reaction, the system was cooled to room temperature and allowed to stand at 0 °C for 12 h to promote solid precipitation. The crude product was collected by vacuum filtration and washed repeatedly with n-hexane to remove residual solvent. The final product was purified by silica gel column chromatography using dichloromethane / methanol (20:1, v / v) as eluent. After solvent removal under reduced pressure, the target product Re-Cy7 was obtained as a dark green crystalline solid with a yield of 75%.

[0040]

[0041] Characterization data of the complex Re-Cy7: 1 H NMR (600MHz, DMSO-d6) δ14.31(s,1H),9.38(d,J=4.7Hz,2H),9.28(s,2H),8.41(d,J=8. 3Hz,2H),8.17(d,J=21.8Hz,2H),7.86(d,J=14.1Hz,2H),7.48(t,J=7.8Hz,4H),7.42–7. 32(m,4H),7.18(t,J=7.3Hz,2H),6.27(d,J=14.2Hz,2H),4.14(s,4H),2.79(s,4H),2.0 1(d,J=10.2Hz,2H),1.74(dd,J=14.3,7.2Hz,4H),1.33(s,12H),0.95(s,6H).FT-IR(KBr pellet,v / cm -1 ):2021,1918,1881(C≡Ostretching),1410-1570(C=C and C=N stretching).ESI-MS(CH3OH):m / z 1121.68[M] + Anal.calcd for C 58 H 55 ClIN6O4Re: C 55.79, H 4.44, N 6.73; found: C 55.75, H 4.48, N6.68. Purity: >96% by HPLC.

[0042] 3. Synthesis of Re-Cy7 NP nanoparticles

[0043] 3 mg of the complex Re-Cy7 was weighed and dissolved in dichloromethane to prepare a homogeneous solution. This solution was then slowly added dropwise to a solution containing 25 mg of Pluronic F-127 and phosphate-buffered saline (PBS) under stirring. The mixture was placed in a fume hood at room temperature and stirred until the dichloromethane was completely evaporated. The resulting mixture was then transferred to a dialysis bag (MWCO, 14000) and dialyzed against deionized water at room temperature for approximately 48 hours to remove small molecule impurities. After dialysis, four ultrafiltration cycles (100 kDa cutoff) were performed to remove excess F-127. The resulting Re-Cy7 nanoparticles were stored in PBS at 4°C for later use.

[0044] Figure 1 The characterization results of Re-Cy7 NPs nanoparticles indicate that uniformly dispersed and stable water-soluble nanoparticles have been successfully synthesized, which can be used for further in vitro and in vivo antibacterial studies.

[0045] Detection of ultrasound-induced ·OH generation

[0046] In the ESR method, 5,5-dimethyl-1-pyrrolline-N-oxide (DMPO) was used as a spin trapping agent to specifically capture ·OH radicals. The reaction solution was prepared by mixing DMPO (25 mM, 20 μL) with Re-Cy7 (100 μM, 80 μL) and adding deionized water, followed by treatment under ultrasonic irradiation conditions (frequency 1.0 MHz, acoustic intensity 1.4 W / cm²). 2 (Duty cycle 50%, 2 minutes). The characteristic signals generated were detected, and the spectral changes were recorded using an electron spin resonance spectrometer to confirm and evaluate the generation of hydroxyl radicals.

[0047] like Figure 2 As shown, the ESR spectrum produced exhibited three distinct characteristic peaks. Among them, the DMPO oxidation product (DMPOX) showed a typical triplet signal, indicating the generation of reactive oxygen species. A clear DMPO-OH quartet signal was also detected, indicating the formation of ·OH. The ESR fitting results were highly consistent with the experimental data, further confirming the formation of ·OH.

[0048] In the MB method, the complex Re-Cy7 was added to a 5 μg / mL methylene blue PBS solution, with a total volume of 3 mL. After equilibration under darkness, the solution was irradiated with ultrasound (frequency 1.0 MHz, sound intensity 1.4 W / cm², duty cycle 50%). Ultraviolet absorption spectra of MB at 663 nm were recorded every 2 minutes for a total of 10 minutes. MB degrades in the presence of ·OH, and the degree of decrease in its absorption peak was used to analyze the amount of ·OH generated.

[0049] The result is as follows Figure 3 As shown, the complex Re-Cy7 can effectively generate ·OH under ultrasonic excitation conditions, producing a ·OH amount comparable to the reference (IR-780), and exhibiting higher stability, indicating that the complex Re-Cy7 has an excellent ability to generate ·OH.

[0050] Ultrasound-induced CO release detection

[0051] The CO release behavior of samples after sonication in PBS buffer solution was detected using the CAY10732 fluorescent probe method. CAY10732 is a CO-specific fluorescent probe with a DMSO stock solution concentration of 2 mM. First, a PBS solution containing 10 μM CAY10732 and PdCl2 was prepared, then 10 μM of the complex Re-Cy7 was added and thoroughly mixed. The sample was then treated under sonication conditions (frequency 1.0 MHz, sound intensity 1.4 W / cm²). 2 The fluorescence signal was collected every 2 minutes at a duty cycle of 50% for a total duration of 10 minutes to monitor the CO release process.

[0052] The results are as follows Figure 4 As shown, the fluorescence intensity gradually increased after ultrasonic treatment, proving that the complex Re-Cy7 can release CO under ultrasonic conditions.

[0053] The CO release capacity was further validated using the hemoglobin method. Hemoglobin (Hb) was dissolved in PBS buffer (4.0 μM, 1 mL, pH 7.4), and reduced by adding sodium sulfite (Na₂S₂O₄, 0.4 mg) under N₂ protection. The complex Re-Cy₇ was then added to the reaction system to a final concentration of 10 μM. Absorption spectra were acquired using a UV-Vis spectrophotometer, and the intensity changes of the absorption peaks at 410 nm (Hb) and 430 nm (HbCO) were recorded. The CO release concentration (C₂CO₃) was calculated according to the Beer-Lambert law. CO ).

[0054] The results are as follows Figure 5 As shown, the Re-Cy7 complex can release CO under US radiation. Further calculations indicate that approximately 3.0 moles of CO are released per mole of Re-Cy7, demonstrating that all three CO ligands in the Re-Cy7 complex can be activated and released. This proves that the Re-Cy7 complex exhibits excellent CO release capability under ultrasonic excitation.

[0055] In vitro antibacterial activity test of Re-Cy7 NPs

[0056] MIC determination: The inhibitory activity of Re-Cy7 NP nanoparticles against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa under ultrasonic conditions was studied using the micro-broth two-fold dilution method. The method was as follows: Bacteria were cultured overnight at 37°C in Mueller-Hinton (MH) broth, and then diluted to 1×10⁻⁶. 6 CFU / mL. 50 μL of the sample solution was added to a 96-well plate for a two-fold serial dilution, followed by 50 μL of diluted bacterial culture. MH broth was used as a positive control. The experimental group was incubated at 37°C for 30 minutes, then subjected to ultrasonic irradiation (1.0 MHz, 1.4 W / cm²) at room temperature. 2 Treat with 50% duty cycle for 2 minutes. Continue incubation at 37°C for 24 hours, then determine the minimum inhibitory concentration (MIC) of the sample.

[0057] The results are shown in Table 1. All tested samples were irradiated with ultrasound (1.0 MHz, 1.4 W / cm²). 2 After 50% duty cycle for 2 minutes, the antibacterial activity was significantly enhanced, with the nanoparticles (Re-Cy7 NPs) showing a particularly significant synergistic effect. Notably, the antibacterial activity of Re-Cy7 NPs against Gram-positive bacteria was more significantly enhanced under ultrasonic stimulation. The antibacterial activity of this material against methicillin-resistant Staphylococcus aureus (MRSA) increased by more than 32 times after ultrasonic irradiation.

[0058] The results demonstrated that Re-Cy7 NP nanoparticles exhibited excellent antibacterial activity under ultrasonic conditions, especially against methicillin-resistant Staphylococcus aureus (MRSA).

[0059] Table 1. Minimum inhibitory concentrations (MICs) of compounds against different bacterial strains

[0060]

[0061] a. Bacteria and drug were incubated in the dark for 24 hours. b. Bacteria were first incubated with the drug in the dark for 30 minutes, then exposed to ultrasound irradiation (1.0 MHz, 1.4 W / cm², 50% cycle, 2 minutes), and then incubated for another 23.5 hours. c. The ultrasonic antibacterial index (SI) was defined as the MIC. Dark / MIC US .

[0062] LB plate count method: Collect the logarithmic growth phase bacterial culture, centrifuge, wash 2-3 times with PBS buffer, and dilute to 3×10⁻⁶. 4CFU / mL. Bacterial suspensions were mixed with different concentrations of drugs and incubated for 30 minutes. The non-ultrasonic group served as a control. The experimental groups were subjected to ultrasonic irradiation (1.0 MHz, 1.4 W / cm²) at 25℃. 2 Treat with 50% duty cycle for 2 minutes. Spread 100 μL of bacterial culture onto LB nutrient agar plates and incubate at 37°C for 18-24 hours before counting single colonies.

[0063] The results are as follows Figure 6 As shown, without ultrasound treatment, even at a concentration of 16 μM, Re-Cy7 NPs showed no significant antibacterial activity. However, under ultrasound irradiation, 16 μM Re-Cy7 NPs reduced the survival rates of the aforementioned pathogens to 1.78% (Staphylococcus aureus), 1.24% (Bacillus subtilis), 2.29% (Escherichia coli), 23.34% (Pseudomonas aeruginosa), and 0.15% (MRSA), respectively. Ultrasound treatment alone had a relatively small impact on bacteria.

[0064] Re-Cy7 NPs in vivo antibacterial activity test

[0065] A skin infection model (MRSA) was established using BALB / c mice to investigate the effects of the complex Re-Cy7 NPs on wound healing and recovery from bacterial infection under ultrasound. Female mice weighing 25–30 g were selected, and their dorsal hair was shaved one day in advance.

[0066] Subsequently, an infection model was established using MRSA: the MRSA bacterial culture was collected by centrifugation, washed twice with PBS, and then diluted to 1.0 × 10⁻⁶ with sterile physiological saline. 7 CFU / mL. An incision of approximately 8mm x 8mm was made on the back using a scalpel, and diluted MRSA solution was immediately instilled to establish infection.

[0067] Twelve hours after infection, mice were randomly divided into four groups (n=5 per group): PBS control group, PBS + ultrasound group, Re-Cy7 NPs group, and Re-Cy7 NPs + ultrasound group. The drug was thoroughly mixed with sterile ointment and applied to the wound surface on the first day of the experiment. The ultrasound treatment groups received ultrasound irradiation (frequency 1.0 MHz, sound intensity 1.4 W / cm²) after drug application. 2 (Duty cycle 50%), lasting 5 minutes.

[0068] Mice were euthanized after treatment, and tissue samples from the infected site were collected and ground. The MRSA bacteria in the tissue samples were rinsed with physiological saline and diluted to 10⁻⁶. 4 After grading, plate culture was performed to count the number of colonies. The area of ​​the infected area, the area of ​​wound healing, and changes in body weight were also recorded.

[0069] Changes in wound area and body weight in mice are as follows: Figure 7 and Figure 8 As shown in the figure. The results indicate that rice-grain Re-Cy7 NPs possess excellent in vivo antibacterial activity. Mice treated with Re-Cy7 NPs and ultrasound showed significantly smaller wound volumes compared to the control group, with a final wound volume of 5.5%. This almost completely inhibited bacterial growth and promoted wound healing, significantly higher than the control group (54.7%). Simultaneously, the mice's body weight did not change significantly, indicating that the complex has minimal toxic side effects. This suggests that Re-Cy7 NPs hold promise as a potential antibiotic alternative for treating multidrug-resistant bacterial infections.

[0070] The above is a further detailed description of the present invention and should not be considered as a limitation on the specific implementation of the present invention. For those skilled in the art, simple deductions or substitutions without departing from the concept of the present invention are all within the protection scope of the present invention.

Claims

1. A nano-formulation of an ultrasonically activated rhenium cyanine dye complex, characterized in that, The preparation method of the rhenium cyanine dye complex nanoparticles includes the following steps: 1) Under strictly anhydrous conditions, the 1,10-o-phenanthroline derivative ligand was deprotonated using sodium hydride, and then the deprotonated product was reacted with IR-780 under an inert gas atmosphere to obtain the intermediate IR780-phen. 2) The complex Re-Cy7 was synthesized by reacting Re(CO)5Cl with IR780-phen under an anhydrous and inert gas atmosphere; 3) The complex Re-Cy7 was added to a solution composed of Pluronic F-127 (F-127) and phosphate buffer (PBS) to finally obtain the rhenium cyanine dye complex nanoparticles Re-Cy7 NPs.

2. The rhenium cyanine dye complex nanoformation according to claim 1, characterized in that, In step 1), the molar ratio of sodium hydride to the 1,10-phenanthroline derivative ligand is 0.9-1.

1.

3. The rhenium cyanine dye complex nanoformation according to claim 1, characterized in that, In step 2), the molar ratio of Re(CO)5Cl to IR780-phen is 0.9-1.

1.

4. The rhenium cyanine dye complex nanoformation according to claim 1, characterized in that, The reaction temperature in step 2) is 110±2℃.

5. The rhenium cyanine dye complex nanoformation according to claim 1, characterized in that, The reaction time for step 2) is 4-5 hours.

6. The rhenium cyanine dye complex nanoformation according to claim 1, characterized in that, The molar ratio of the deprotonated product to IR-780 in step 1) is 1.9-2.

1.

7. The use of the rhenium cyanine dye complex nanoformation according to claim 1 in the preparation of formulations for treating bacterial infections and promoting wound healing.

8. The application according to claim 7, characterized in that, The preparation is a preparation for sonodynamic therapy.

9. The application according to claim 7, characterized in that, The preparation is a gas therapy preparation.

10. The application according to claim 7, characterized in that, The preparation is a therapeutic preparation for sonodynamic therapy combined with gas therapy.

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