Photo-thermal agent for treating allergic rhinitis and application of photo-thermal agent in preparation of medicine for treating allergic rhinitis

Abstract

The invention discloses a photo-thermal agent for treating allergic rhinitis and application of the photo-thermal agent in preparation of drugs for treating allergic rhinitis, a method comprises the following steps: firstly, reacting a compound 1 with two pyridine groups with diethyleneglycol-2-bromoethyl methyl ether to prepare an ether-oxygen chain substituted compound 2, then dissolving the prepared compound 2 and DSPE-mPEG5000 in tetrahydrofuran, and carrying out a reaction to obtain the photo-thermal agent for treating allergic rhinitis. According to the invention, the pyridinium salt group is taken as a carrier, self-assembly is carried out to form nano-particles, and the nano-particles promote intramolecular charge transfer and cause red shift of absorption wavelength due to the strong electron withdrawing effect of the pyridinium salt group, have deeper tissue penetrating power, and can effectively reach the deep part of nasal mucosa to treat deep inflammation; the RH-coated NPs prepared by the invention has both high-efficiency photothermal conversion performance and photodynamic therapy sensitivity, so that the synergistic effect of PTT and PDT on treatment of allergic rhinitis becomes possible, the treatment effect is greatly improved, and a new way is opened up for treatment of allergic rhinitis.

Description

Technical Field

[0001] The present invention belongs to the technical field of photothermal therapy, and relates to a photothermal agent for treating allergic rhinitis and application of the photothermal agent in preparing a drug for treating allergic rhinitis. Background Art

[0002] Allergic rhinitis is one of the most common chronic diseases in the world, with a global prevalence of 20% to 30%, and has been on the rise for many years. Allergic rhinitis is an IgE-mediated nasal allergic reaction caused by inhaled allergens, manifested by sneezing, nasal congestion, nasal itching and runny nose. Clinically, drug therapy is the most commonly used treatment method, but its treatment effect varies greatly from person to person. According to statistics, up to 29% of children and 62% of adults only partially relieve or do not relieve their symptoms after receiving drug treatment alone, and long-term medication can cause local symptoms such as local irritation and bleeding, as well as systemic adverse reactions such as drowsiness and fatigue. At present, there is an urgent need for effective new treatments that can target local areas to relieve local symptoms without causing systemic adverse reactions.

[0003] As an emerging, safe and non-invasive treatment method, phototherapy shows broad application prospects in the treatment of diseases by producing photothermal effect (PTT) and photodynamic effect (PDT). PTT has the advantages of high efficiency, controllability and low toxicity. Mild PTT can regulate immunity and epithelial-mesenchymal transition (EMT) under inflammatory conditions. PDT, as an effective photodynamic process, can activate the production of reactive oxygen species (ROS) through light irradiation. Reactive oxygen can serve as cell signaling molecules to maintain redox balance and promote health. Based on this characteristic, PDT has been widely studied in the treatment of other diseases, but there are few reports on the synergistic treatment of allergic rhinitis using PTT and PDT. In addition, current photothermal agents mainly focus on phototherapy using light bands within the ultraviolet / visible spectrum, while ignoring the use of near-infrared light (NIR), especially NIR-II region light with a wavelength range of 1000 to 1700 nm, which is potentially effective in the treatment of rhinitis; and the amount of ROS generated by photothermal agents in the prior art is uncontrollable, but a high content of ROS can kill cells; at the same time, due to the obstruction of nasal bones and soft tissues, during treatment, it is necessary to adjust the material properties and screen the light power to achieve the ideal penetration depth while maintaining good biosafety. Summary of the invention

[0004] In view of the problems existing in the prior art, the present invention provides a photothermal agent for treating allergic rhinitis and use thereof in preparing a drug for allergic rhinitis, thereby solving the technical problem that the single PTT method used in the prior art cannot effectively achieve sufficient treatment of allergic rhinitis, and the existing photothermal agent cannot effectively utilize NIR-II region light during PDT treatment, and the amount of ROS generated is uncontrollable, and the technical problem that the effect of PDT therapy in the treatment of allergic rhinitis cannot be fully exerted.

[0005] The present invention is achieved through the following technical solutions:

[0006] A method for preparing a photothermal agent for treating allergic rhinitis comprises the following steps:

[0007] S1: under nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether was added to a chloroform solution of compound 1, stirred for reaction, and then concentrated and purified to obtain compound 2;

[0008] The structural formula of the compound 1 is:

[0009]

[0010] S2: Compound 2 and DSPE-mPEG 5000 The RH@NPs were dissolved in tetrahydrofuran and mixed evenly by ultrasonication. The mixed solution was then injected into water to disperse the system evenly. The tetrahydrofuran was then evaporated under a nitrogen atmosphere and filtered to obtain the RH@NPs-based NIR-II region photothermal agent.

[0011] Preferably, in step S1, during the reaction process, the reaction temperature is 60-90° C. and the reaction time is 12-24 h.

[0012] Preferably, in step S1, the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether is 1:(1-2).

[0013] Preferably, in step S1, vacuum concentration is used for concentration treatment, and during the vacuum concentration treatment, the vacuum degree is -0.05 to -0.1 MPa.

[0014] Preferably, in step S1, purification is performed by chromatography column purification, and during the chromatography column purification process, the stationary phase is silica gel, and the mobile phase is a mixture of methanol and dichloromethane in a volume ratio of 1:25.

[0015] Preferably, in step S2, the compound 2 and DSPE-mPEG 5000 The mass ratio is 1:(5~10).

[0016] Preferably, in step S2, the temperature for evaporating and removing tetrahydrofuran under a nitrogen atmosphere is 22 to 25° C. and the time is 18 to 24 hours.

[0017] Preferably, in step S2, the pore size of the filter membrane used for filtration is 0.2-0.45 μm.

[0018] A photothermal agent for treating allergic rhinitis is prepared by the above method; the average size of the NIR-II region photothermal agent based on RH@NPs is 100-160nm, and the molar absorption coefficient of the photothermal agent in the NIR-II region is 4000-4300M -1 cm -1 ).

[0019] The use of the above-mentioned photothermal agent for treating allergic rhinitis in the preparation of allergic rhinitis medicine.

[0020] Compared with the prior art, the present invention has the following beneficial technical effects:

[0021] The invention discloses a preparation method of a photothermal agent for treating allergic rhinitis. Firstly, the photothermal agent prepared by the method has both high-efficiency photothermal conversion performance and sensitivity of photodynamic therapy, so that photothermal therapy (PTT) and photodynamic therapy (PDT) synergistically treat allergic rhinitis possible, greatly improving the treatment effect, and opening up a new way for the treatment of allergic rhinitis. PTT (photothermal therapy) utilizes a photothermal agent to absorb light energy of a specific wavelength and convert it into heat energy, gently raising the local temperature, helping to relieve swelling and congestion of the nasal mucosa, and reducing allergic reactions, while PDT (photodynamic therapy) mobilizes the antioxidant system of the mucosa through the active oxygen species generated by the photosensitizer under light, and at the same time regulates the immune response and reduces the release of inflammatory mediators. The combination of the two therapies not only improves the nasal environment from a physical level, but also regulates the body's immune response from the root, providing a more comprehensive, efficient and less side-effect treatment plan for patients with allergic rhinitis. In addition, in the preparation process of the photothermal agent of the present invention, the compound 1 with two pyridine groups is firstly reacted with diethylene glycol-2-bromoethyl methyl ether to obtain the ether oxygen chain-substituted compound 2. In this process, the pyridinium salt group acts as a strong electron attracting group, which enhances the push-pull electron effect, promotes the charge transfer within the molecule, and leads to the red shift of the absorption wavelength, which can improve the penetration depth of rhinitis treatment and reduce tissue light damage. In addition, the ether oxygen chain is used to replace the pyridine group, which enhances the electron attraction ability, while maintaining the self-assembly ability, promotes the coupling between excited states, and is beneficial to the photodynamic behavior. At the same time, in the process of ether oxygen chain substitution, diethylene glycol-2-bromoethyl methyl ether is added to the chloroform solution of compound 1 by controlling the dropwise addition order, so as to achieve the unilateral pyridine substitution of compound 1. The unilateral pyridine substitution transforms compound 1 from the symmetrical C2v point group DA structure to the asymmetrical C1 point group DA RH structure, increases the molecular polarity and dipole moment, promotes the self-assembly of compound 2, enhances the inter-system cross-ability, and realizes an efficient photodynamic process. Then, the compound 2 and DSPE-mPEG 5000Compound 2 was dissolved in tetrahydrofuran to self-assemble into nanoparticles. The self-assembly process made the interaction between molecules closer, which was conducive to the energy transfer and coupling between excited state molecules. Because in the photodynamic process, the interaction between excited state molecules is the key to the photodynamic effect. Through self-assembly, more excited state molecules can be placed in similar spatial positions, thereby increasing the chance of interaction between them and improving the efficiency of the photodynamic effect. The self-assembled nanoparticles promote the intramolecular charge transfer due to the strong electron-withdrawing effect of the pyridinium salt group, resulting in a red shift in the absorption wavelength, so that they can better respond to long-wavelength light with high tissue penetration ability, thereby penetrating the nasal bone and soft tissue to treat nasal mucosal inflammation. At the same time, due to the substitution of the ether oxygen chain on the nanoparticle, the electron attraction ability is enhanced, showing a moderate photothermal conversion efficiency, and can quickly heat up to the ideal treatment temperature under laser irradiation, while maintaining good thermal stability, producing a good PTT effect. In addition, the nanoparticles can generate superoxide free radicals (·O2 - ), and the lifetime of the excited state in the near-infrared zone II is short, so the free radicals generated are at a relatively low level, and can be used as cell signaling molecules at low doses to trigger antioxidant defense, promote health, avoid cell damage that may be caused by overtreatment, and achieve a good PDT effect; moreover, in in vitro experiments, even at higher concentrations, RH@NPs have little effect on the survival rate of nasopharyngeal epithelial cells, showing good biocompatibility, and RH@NPs can be quickly enriched in the nasal cavity and quickly cleared after laser irradiation, reducing the side effects that may be caused by long-term retention. The preparation method of the present invention is simple and reasonably designed. The prepared RH@NPs have good PTT and PDT dual effects, and have good penetration depth, which effectively improves the long-term health improvement of the nasal mucosa.

[0022] Furthermore, in the step S1, during the temperature-raising reaction, the reaction temperature is 60 to 90° C., and the reaction time is 12 to 24 hours. Appropriate temperature can increase the rate of the chemical reaction, allowing the reactants to contact and react more fully. Within the temperature range of 60 to 90° C., the reaction rate is moderate, which can not only ensure the progress of the reaction, but also avoid unnecessary side reactions or product degradation caused by excessively high temperatures. Sufficient reaction time allows the reactants to react fully, thereby improving the yield and purity of the product.

[0023] Furthermore, in step S1, the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether is 1:(1-2). An appropriate molar ratio can ensure sufficient contact and reaction between the reactants, thereby improving the reaction efficiency. When the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether is controlled within the range of 1:(1-2), it can ensure that there is sufficient diethylene glycol-2-bromoethyl methyl ether to react with compound 1 to generate the target product compound 2. This molar ratio setting helps to reduce unreacted raw materials, thereby improving the yield and purity of the product.

[0024] Furthermore, in the step S1, a vacuum concentration method is used for concentration treatment. During the vacuum concentration treatment, the vacuum degree is -0.05 to -0.1 MPa, which can completely evaporate the solvent contained in the RH@NPs, and obtain RH@NPs with higher purity.

[0025] Furthermore, in step S1, purification is performed by chromatography column purification. During the chromatography column purification process, the stationary phase is silica gel, and the mobile phase is a mixture of methanol and dichloromethane in a volume ratio of 1:25, which can shorten the purification time and efficiently obtain a high-purity RH@NPs photothermal agent.

[0026] Further, in step S2, the compound 2 and DSPE-mPEG 5000 The mass ratio of 1:(5~10) can obtain RH@NPs with uniform morphology and appropriate size, and the size of the material has a significant effect on its stability and cellular uptake ability.

[0027] Furthermore, in the step S2, the temperature for evaporating and removing tetrahydrofuran under a nitrogen atmosphere is 22 to 25° C. and the time is 18 to 24 hours, so that stable RH@NPs can be obtained.

[0028] Furthermore, in step S2, the pore size of the filter membrane used for filtration is 0.2-0.45 μm, so that large nanoparticles that are not suitable for biological applications can be excluded. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.

[0030] Figure 1The synthesis process and photophysical property test results of the photothermal agent RH@NPs prepared in Example 1 of the present invention, wherein: (a) is the self-assembly process of RH@NPs; (b) is the comparison of the absorption and emission spectra of compound 1 (SW6) and RH; (c) is the absorption and emission spectra of RH (in DMSO, concentration = 10 -5 mol / L,λ ex =808nm); (d) is the absorption spectrum of RH (ε: molar absorption coefficient); (e) is the average diameter of RH@NPs shown by transmission electron microscopy (TEM) and dynamic light scattering (DLS); (f) is the absorption and emission spectra of RH@NPs (in H2O, concentration = 10 -5 mol / L,λ ex =808nm); (g) is the absorption spectrum of RH@NPs (ε: molar absorption coefficient); (h) is the photothermal heating curve of RH@NPs;

[0031] Figure 2 This is a free radical capture test of the RH@NPs photothermal agent prepared in Example 1 of the present invention;

[0032] Figure 3 In vitro cell experiments of the photothermal agent RH@NPs prepared in the present invention: (a) CCK8 experiment to evaluate the different RH@NPs concentrations (0, 0.5, 1, 2 mg mL -1 ) is the viability of NP69 cells; (b) is the fluorescence microscopy detection of the uptake of RH@NPs by human nasopharyngeal epithelial NP69 cells; (c) is the fluorescence quantitative graph of Figure (b)

[0033] Figure 4 The enrichment, maintenance and clearance of the photothermal agent RH@NPs prepared in the present invention in the nasal mucosa of mice: the enrichment, maintenance and clearance of RH@NPs in the nasal mucosa were evaluated in a mouse model by nasal spray administration, and the changes in nasal temperature after laser irradiation were monitored using infrared thermal imaging technology;

[0034] Figure 5 The photothermal treatment of allergic rhinitis in mice by the photothermal agent RH@NPs prepared in the present invention: wherein, (a) is a flowchart of the in vivo experiment; (b) is a gross behavioral evaluation of mice with allergic rhinitis; (c) is the content of serum histamine and allergen-specific IgE; (d) is the result of pathological staining of nasal mucosal tissue sections of mice with allergic rhinitis;

[0035] Figure 6 The therapeutic effect of the photothermal agent RH@NPs prepared in the present invention on oxidative stress and inflammation in mice with allergic rhinitis: wherein, (a) is the level of nasal mucosal inflammatory factors; (b) is the level of nasal mucosal glutathione anabolism-related factors; (c) is the mRNA level of nasal mucosal EMT-related factors. DETAILED DESCRIPTION

[0036] In order to enable those skilled in the art to understand the characteristics and effects of the present invention, the following is a general description and definition of the terms and expressions mentioned in the specification and claims. Unless otherwise specified, all technical and scientific terms used in the text are the common meanings understood by those skilled in the art for the present invention. In the event of a conflict, the definition in this specification shall prevail.

[0037] The theories or mechanisms described and disclosed herein, whether correct or incorrect, should not limit the scope of the present invention in any way, that is, the present invention can be implemented without being limited by any specific theory or mechanism.

[0038] Herein, all features such as values, quantities, contents and concentrations defined in the form of numerical ranges or percentage ranges are for simplicity and convenience only. Accordingly, the description of numerical ranges or percentage ranges should be considered to have included and specifically disclosed all possible secondary ranges and individual values ​​within the range (including integers and fractions).

[0039] In this document, unless otherwise specified, “includes,” “including,” “contains,” “has,” or similar terms cover the meanings of “consisting of” and “mainly consisting of,” for example, “A includes a” covers the meanings of “A includes a and other” and “A only includes a.”

[0040] In this document, in order to make the description concise, not all possible combinations of various technical features in various embodiments or examples are described. Therefore, as long as there is no contradiction in the combination of these technical features, the various technical features in various embodiments or examples can be combined arbitrarily, and all possible combinations should be considered to be within the scope of this specification.

[0041] The present invention provides a method for preparing a photothermal agent for treating allergic rhinitis, comprising the following steps:

[0042] S1: Under nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether is added to the chloroform solution of compound 1, wherein the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether is 1:(1-2), after stirring evenly, the temperature is raised to react the system at 60-90°C for 12-24h, and the system is concentrated by vacuum concentration at a vacuum degree of -0.05-0.1MPa, and purified by chromatography column purification, wherein the stationary phase is silica gel, and the mobile phase is a mixture of methanol and dichloromethane in a volume ratio of 1:25, and after the purification treatment is completed, compound 2 is obtained. In this step, diethylene glycol-2-bromoethyl methyl ether is added three times, each time adding one-third of the calculated amount, stirring for 30min, and then adding the next batch of diethylene glycol-2-bromoethyl methyl ether.

[0043] The structural formula of the compound 1 is:

[0044]

[0045] The structural formula of the obtained compound 2 is:

[0046]

[0047] The preparation process of the above-mentioned compound 1 is based on the reference patent CN116606313A.

[0048] S2: Figure 1 As shown in (a), the compound 2 and phospholipid methoxy polyethylene glycol (DSPE-mPEG 5000 ) are dissolved in tetrahydrofuran in a mass ratio of 1:(5-10), ultrasonically mixed, and then the mixture is injected into water, the system is evenly dispersed under a cell disruptor, and then the system is placed in a nitrogen atmosphere, stirred at 22-25°C for 18-24h, the tetrahydrofuran is evaporated, and filtered with a filter membrane with a pore size of 0.2-0.45μm to obtain a RH@NPs-based photothermal agent for the NIR-II region.

[0049] In the present invention, 1-(2-Bromoethoxy)-2-(2methoxyethoxy)ethane is synthesized by a nucleophilic substitution reaction with the pyridine group in compound 1 to form RH with a pyridinium salt structure, i.e., compound 2. In this process, the pyridinium salt group is introduced as a strong electron-attracting group, which enhances the push-pull electron effect, promotes the charge transfer within the molecule, leads to a red shift in the absorption wavelength, improves the penetration depth of rhinitis treatment, and reduces tissue photodamage. The use of ether oxygen chains instead of alkyl chains enhances the electron-attracting ability, while maintaining the self-assembly ability, promotes the coupling between excited states, and is beneficial to photodynamic behavior. The unilateral pyridine substitution transforms compound 1 from a symmetrical C2v point group DA structure to an asymmetric C1 point group D-ARH structure, increases the molecular polarity and dipole moment, promotes the self-assembly of RH, enhances the inter-system crossover ability, and achieves efficient photodynamic generation. In addition, the prepared RH@NPs showed photothermal conversion ability under 808nm and 1064nm laser irradiation. Among them, 1064nm laser is more suitable for treating deep nasal inflammation due to its deeper tissue penetration ability. RH@NPs has a moderate photothermal conversion efficiency (34%) and can produce superoxide anions (·O2 - ), which is the main reactive oxygen species produced by the photodynamic effect. At the same time, RH@NPs exhibited excellent in vitro biocompatibility, and even at a concentration as high as 2 mg / mL, it had little effect on the survival rate of the nasopharyngeal epithelial cell line NP69. Through nasal spray administration, RH@NPs can quickly enrich in the nasal mucosa, reach the ideal photothermal temperature after laser irradiation, and are effectively cleared within a few hours, reducing side effects caused by systemic absorption. In addition, in the OVA-induced AR mouse model, nasal spray administration of RH@NPs combined with laser irradiation significantly alleviated the acute symptoms of AR, reduced allergic mediators such as histamine and OVA-specific IgE in serum, and improved epithelial damage to the nasal mucosa.

[0050] The present invention also discloses a photothermal agent for treating allergic rhinitis prepared by the above method, wherein the average size of the NIR-II region photothermal agent based on RH@NPs is 100-160 nm, and the molar absorption coefficient of the photothermal agent in the NIR-II region is (4000-4300 M -1 cm -1 ).

[0051] The present invention also explores the photothermal and photodynamic properties of RH@NPs at different concentrations and laser power densities, as well as the therapeutic effects in different animal models, providing the possibility of multiple application scenarios and technical adjustments. The technical solution of the present invention is innovative and provides a safe, effective and controllable new treatment method for allergic rhinitis. Through precise molecular design and regulation of photophysical properties, RH@NPs can achieve efficient PTT and PDT effects in the NIR-II region, providing a new strategy for the treatment of allergic rhinitis.

[0052] The present invention proposes a photothermal agent RH@NPs for synergistic treatment of allergic rhinitis using photothermal effect (PTT) and photodynamic effect (PDT). The photothermal agent has the following advantages:

[0053] (1) Deep tissue penetration: RH@NPs are specifically designed to respond to near-infrared II (NIR-II) light, which has deeper tissue penetration.

[0054] (2) High photothermal conversion efficiency: RH@NPs exhibit moderate photothermal conversion efficiency and can quickly heat up to the ideal therapeutic temperature under laser irradiation while maintaining good thermal stability.

[0055] (3) Precise ROS production: RH@NPs can produce specific types of reactive oxygen species (O2 - ·) At low doses, it can act as a cell signaling molecule to trigger antioxidant defense and promote health. At the same time, the amount of free radicals produced can be controlled by controlling the irradiation time and power. At the same time, the concentration of free radicals produced by near-infrared zone II materials is relatively low, which can also effectively avoid cell damage that may be caused by excessive treatment.

[0056] (4) Excellent biocompatibility: In in vitro experiments, even at higher concentrations, RH@NPs had little effect on the survival rate of nasopharyngeal epithelial cells, showing good biocompatibility.

[0057] (5) Rapid clearance in vivo: RH@NPs can be rapidly enriched in the nasal cavity and cleared quickly after laser irradiation, reducing the side effects that may be caused by long-term retention.

[0058] (6) Comprehensive therapeutic effect: Through phototherapy, RH@NPs can not only relieve the acute symptoms of allergic rhinitis, but also improve the long-term health of the nasal mucosa by enhancing antioxidant capacity and inhibiting the epithelial-mesenchymal transition (EMT) process.

[0059] (7) Potential clinical application prospects: The RH@NPs of the present invention provide a safe, effective and controllable new option for the clinical treatment of allergic rhinitis, which is expected to reduce the economic burden of patients and improve their quality of life. The RH@NPs of the present invention provide new ideas and methods for the application of phototherapy in the treatment of other diseases, and promote scientific research and technological development in related fields.

[0060] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the appended claims of the application equally.

[0061] The following examples use conventional instruments and equipment in the art. The experimental methods in the following examples where specific conditions are not specified are usually carried out under conventional conditions or under conditions recommended by the manufacturer. The various raw materials used in the following examples are conventional commercial products unless otherwise specified, and their specifications are conventional specifications in the art. In the specification of the present invention and the following examples, unless otherwise specified, "%" means weight percentage, "part" means weight part, and ratio means weight ratio.

[0062] Example 1

[0063] A method for preparing a photothermal agent for treating allergic rhinitis comprises the following steps:

[0064] S1: Under N2 protection, compound 1 (50 mg, 0.06 mmol) and diethylene glycol-2-bromoethyl methyl ether (27.3 mg, 0.12 mmol) were added to a round-bottom flask filled with CHCl3 (10 mL), stirred at 90°C for 24 h, the reaction was cooled to room temperature, and the solvent was concentrated in vacuo. Finally, RH was purified by chromatography column (silica gel, MeOH / DCM=1 / 25) to obtain RH as a blue-green solid (yield 20%), i.e., compound 2, denoted as RH.

[0065] 1 H NMR(500MHz,CDCl3)δ / ppm 9.26(s,2H),8.63(s,2H),8.12(s,6H),7.67(t,J=15.0,10H),6.79(s,2H),5. 00(s,2H),4.03(s,2H),3.56(s,8H),3.32(s,11H),2.75(s,8H),1.98(s,8H). 13C NMR (126MHz, CDCl3) δ / ppm155.7,150.4,147.6,145.5,139.2,133.5,130.6,130.2,127.7,127.0,124.2,121.6,1 14.5,77.2,72.0,70.6,70.5,70.3,69.5,60.5,59.2,50.3,32.1,29.8,28.2,22.8,21.8,14.2.MALDI-TOF / MS:[M] + calcd:989.011,found:988.941.

[0066] S2: 1 mg of compound 2 and 5 mg of DSPE-mPEG 5000 The obtained product was dissolved in 1 mL of tetrahydrofuran and mixed evenly by ultrasonication. The mixture was then injected into 9 mL of deionized water to disperse the system evenly. The system was then placed under a nitrogen atmosphere and stirred at 22 °C for 18 h. The tetrahydrofuran was evaporated and filtered through a filter membrane with a pore size of 0.45 μm to obtain a RH@NPs-based photothermal agent for the NIR-II region.

[0067] The RH and RH@NPs prepared in this embodiment are used to illustrate the technical effects of the present invention, specifically:

[0068] Figure 1 The synthesis process of the photothermal agent RH@NPs in the present invention and the test results of the photophysical properties of RH and RH@NPs synthesized in Example 1, wherein: (a) is the self-assembly process of RH@NPs; (b) is the comparison of the absorption spectrum and emission spectrum of compound 1 (SW6) and RH; (c) is the absorption spectrum and emission spectrum of RH (in dimethyl sulfoxide (DMSO), concentration = 10 -5 mol / L,λ ex =808nm); (d) is the absorption spectrum of RH (ε: molar absorption coefficient); (e) is the average diameter of RH@NPs shown by transmission electron microscopy (TEM) and dynamic light scattering (DLS); (f) is the absorption and emission spectra of RH@NPs (in H2O, concentration = 10 -5 mol / L,λ ex =808nm); (g) is the absorption spectrum of RH@NPs (ε: molar absorption coefficient); (h) is the photothermal heating curve of RH@NPs. As can be seen from the figure, the RH molecule introduces the pyridine cyanide unit as a strong electron-withdrawing group, which enhances the push-pull electron effect and the electron delocalization ability. Compared with compound 1, the red shift of the absorption wavelength of RH can improve the penetration depth of the treatment. Figure 1(b), (c), and (d) show that the main absorption and emission peaks (998nm) of RH molecules are closer to 1000nm. Compared with RH molecules, the absorption spectrum of RH@NPs nanoparticles is significantly broader than that of organic molecules, indicating the coexistence of multiple aggregation states, and after being assembled into nanoparticles, its absorption spectrum has a slight blue shift, indicating that there is a Coulomb coupling effect in the molecules in the nanoparticles after coating, and in the process of material construction, only by first red-shifting to a longer wavelength and then blue-shifting can the penetration depth of RH@NPs nanoparticles be guaranteed. Figure 1 (e) It can be seen that a spherical nanostructure with a uniform shape and a size of 100 nm is formed; Figure 1 (h) It can be seen that RH@NPs nanoparticles have good photothermal conversion ability;

[0069] Figure 2 The free radical capture test of the RH@NPs photothermal agent prepared in Example 1 of the present invention is performed by using 2',7'-dichlorodihydroxyfenol (DCFH) as a fluorescent probe, and adding RH@NPs photothermal agent and different active oxygen scavengers to perform fluorescence quenching experiments. The active oxygen scavengers are NaN3 (SA, used for 1 O2), vitamin C (VC, used for O2 - ·) and isopropyl alcohol (IPA, for ·OH), test results are shown in Figure 2 As shown by Figure 2 It can be seen that only after adding VC, the fluorescence intensity was significantly inhibited, indicating that RH@NPs produced O2 under both 808nm and 1064nm light. - ·, with type I photodynamic therapy capability.

[0070] Figure 3 In vitro cell experiments of the photothermal agent RH@NPs prepared in the present invention: (a) CCK8 experiment to evaluate the different RH@NPs concentrations (0, 0.5, 1, 2 mg mL -1 ) shows the viability of NP69 cells; (b) is the fluorescence microscopy test of the uptake of RH@NPs by human nasopharyngeal epithelial NP69 cells, and (c) is the fluorescence quantitative graph of (b). Figure 3 (a) It can be seen that RH@NPs have good biocompatibility, do not affect cell proliferation, and have no cytotoxicity; Figure 3 (b) to (c) show that RH@NPs can be effectively taken up by nasopharyngeal epithelial cells and quickly reach an ideal enrichment state.

[0071] Figure 4The enrichment, maintenance and clearance of the photothermal agent RH@NPs prepared in Example 1 of the present invention in the nasal mucosa of mice, specifically, the enrichment, maintenance and clearance of RH@NPs in the nasal mucosa were evaluated in a mouse model by nasal spray administration, and the changes in nasal temperature after laser irradiation were monitored using infrared thermal imaging technology. It can be seen from the figure that compared with the blank control (PBS group), the nasal temperature of the RH@NPs group rose rapidly to 39.1°C within 4 minutes and maintained for at least 20 minutes, and there was no obvious swelling of the nasal mucosa, and it was shown that the photothermal effect mediated by RH@NPs has the characteristics of rapid, stable and moderate. The highest temperature in the nasal cavity dropped within 3 hours and returned to normal within about 6 hours. This temperature change indicates that RH@NPs are rapidly cleared from the nasal mucosa through nasal ciliary oscillation and mucus barrier.

[0072] Figure 5 The photothermal treatment of allergic rhinitis in mice by the photothermal agent RH@NPs prepared in Example 1 of the present invention: (a) is a flowchart of the in vivo experiment; (b) is a gross behavioral evaluation of mice with allergic rhinitis, wherein the left figure is the frequency data of sneezing in diseased mice, and the right figure is the frequency data of scratching the nose in diseased mice; (c) is an evaluation of the level of allergic mediators in the serum of mice with allergic rhinitis, wherein the left figure is the change in the content of histamine in the blood of diseased mice, and the right figure is the change in the content of allergen-specific IgE in the blood of diseased mice; (d) The results of pathological staining of nasal mucosal tissue sections of mice with allergic rhinitis, among which, HE staining was used to quantitatively detect the thickness of epithelial cells in the respiratory zone of the nasal septum to evaluate nasal mucosal damage, and PAS staining was used to evaluate the hyperplasia of nasal mucosal goblet cells. As can be seen from the figure, RH@NPs treatment via nasal spray has a good therapeutic effect on mice with allergic rhinitis, which is manifested in the reduction of allergic symptoms of sneezing and scratching in diseased mice, reduction of histamine and allergen-specific IgE in the blood, reduction of swelling damage of the nasal septum mucosa and reduction of goblet cell hyperplasia.

[0073] Figure 6 The therapeutic effect of the photothermal agent RH@NPs prepared in Example 1 of the present invention on oxidative stress and inflammation in mice with allergic rhinitis: wherein, (a) is the level of nasal mucosal inflammatory factors; (b) is the level of nasal mucosal glutathione anabolism-related factors; (c) is the mRNA level of nasal mucosal EMT-related factors. As can be seen from the figure, RH@NPs combined with NIR-II zone light irradiation can reduce the expression of type 2 inflammation-related genes such as Il4, Il5, Il3 and Gata3 in the nose of mice with allergic rhinitis, upregulate the expression of antioxidant genes such as Gclc, Gclm and Gpx2, and downregulate the expression of EMT genes such as Col11a1 and Col3a1, thereby enhancing the antioxidant capacity of the nasal mucosa and inhibiting EMT while reducing inflammation.

[0074] Example 2

[0075] A method for preparing a photothermal agent for treating allergic rhinitis comprises the following steps:

[0076] S1: Under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether was added to a chloroform solution of compound 1 in three batches. After each addition of diethylene glycol-2-bromoethyl methyl ether, the mixture was stirred for 30 minutes before adding the next batch of diethylene glycol-2-bromoethyl methyl ether, wherein the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether was 1:1. After stirring evenly, the system was heated to react at 60°C for 24 hours. The system was concentrated by vacuum concentration at a vacuum degree of -0.05 MPa, and purified by chromatography column purification. In the chromatography column purification process, silica gel was used as the stationary phase, and a mixture of methanol and dichloromethane in a volume ratio of 1:25 was used as the mobile phase. After the purification process was completed, compound 2 was obtained.

[0077] The structural formula of the compound 1 is:

[0078]

[0079] S2: Compound 2 and phospholipid methoxypolyethylene glycol (DSPE-mPEG 5000 ) were dissolved in tetrahydrofuran at a mass ratio of 1:5 and ultrasonically mixed. The mixture was then injected into water, and the system was evenly dispersed under a cell disruptor. The system was then placed in a nitrogen atmosphere and stirred at 22°C for 24 hours. The tetrahydrofuran was evaporated and filtered through a filter membrane with a pore size of 0.2 μm to obtain a RH@NPs-based photothermal agent for the NIR-II region.

[0080] Example 3

[0081] A method for preparing a photothermal agent for treating allergic rhinitis comprises the following steps:

[0082] S1: Under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether was added to a chloroform solution of compound 1 in three batches. After each addition of diethylene glycol-2-bromoethyl methyl ether, the mixture was stirred for 30 minutes before adding the next batch of diethylene glycol-2-bromoethyl methyl ether, wherein the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether was 1:2. After stirring evenly, the system was heated to react at 90°C for 12 hours. The system was concentrated by vacuum concentration at a vacuum degree of -0.1 MPa, and purified by chromatography column purification. In the chromatography column purification process, silica gel was used as the stationary phase, and a mixture of methanol and dichloromethane in a volume ratio of 1:25 was used as the mobile phase. After purification, compound 2 was obtained.

[0083] The structural formula of the compound 1 is:

[0084]

[0085] S2: Compound 2 and phospholipid methoxypolyethylene glycol (DSPE-mPEG 5000 ) were dissolved in tetrahydrofuran at a mass ratio of 1:(5-10), and ultrasonically mixed. The mixture was then injected into water, and the system was evenly dispersed under a cell disruptor. The system was then placed in a nitrogen atmosphere and stirred at 25°C for 18 hours. The tetrahydrofuran was evaporated and filtered through a filter membrane with a pore size of 0.45 μm to obtain a RH@NPs-based photothermal agent for the NIR-II region.

[0086] Example 4

[0087] A method for preparing a photothermal agent for treating allergic rhinitis comprises the following steps:

[0088] S1: Under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether was added to a chloroform solution of compound 1 in three batches. After each addition of diethylene glycol-2-bromoethyl methyl ether, the mixture was stirred for 30 minutes before adding the next batch of diethylene glycol-2-bromoethyl methyl ether, wherein the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether was 1:1.5. After stirring evenly, the system was heated to react at 80°C for 20 hours. The system was concentrated by vacuum concentration at a vacuum degree of -0.1 MPa, and purified by chromatography column purification. In the chromatography column purification process, silica gel was used as the stationary phase, and a mixture of methanol and dichloromethane in a volume ratio of 1:25 was used as the mobile phase. After the purification process was completed, compound 2 was obtained.

[0089] The structural formula of the compound 1 is:

[0090]

[0091] S2: Compound 2 and phospholipid methoxypolyethylene glycol (DSPE-mPEG 5000 ) were dissolved in tetrahydrofuran at a mass ratio of 1:8 and ultrasonically mixed. The mixture was then injected into water, and the system was evenly dispersed under a cell disruptor. The system was then placed in a nitrogen atmosphere and stirred at 23°C for 20 hours. The tetrahydrofuran was evaporated and filtered through a filter membrane with a pore size of 0.3 μm to obtain a RH@NPs-based photothermal agent for the NIR-II region.

[0092] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing a photothermal agent for treating allergic rhinitis, characterized in that: The following steps are involved: S1: under nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether was added to a chloroform solution of compound 1, stirred for reaction, and then concentrated and purified to obtain compound 2; The structural formula of the compound 1 is: S2: Compound 2 and DSPE-mPEG 5000 The mixture is dissolved in tetrahydrofuran and uniformly mixed by ultrasonication, and then the mixture is injected into water to disperse the system uniformly, and then the tetrahydrofuran is evaporated under a nitrogen atmosphere and filtered to obtain the photothermal agent for treating allergic rhinitis.

2. The method for preparing a photothermal agent for treating allergic rhinitis according to claim 1, characterized in that: In the step S1, during the reaction, the reaction temperature is 60-90° C. and the reaction time is 12-24 hours.

3. The method for preparing a photothermal agent for treating allergic rhinitis according to claim 1, characterized in that: In the step S1, the molar ratio of the compound 1 to diethylene glycol-2-bromoethyl methyl ether is 1:(1-2).

4. The method for preparing a photothermal agent for treating allergic rhinitis according to claim 1, characterized in that: In the step S1, vacuum concentration is used for concentration treatment. During the vacuum concentration treatment, the vacuum degree is -0.05 to -0.1 MPa.

5. The method for preparing a photothermal agent for treating allergic rhinitis according to claim 1, characterized in that: In the step S1, purification is performed by chromatography column purification. During the chromatography column purification process, the stationary phase is silica gel, and the mobile phase is a mixture of methanol and dichloromethane in a volume ratio of 1:

25.

6. The method for preparing a photothermal agent for treating allergic rhinitis according to claim 1, characterized in that: In the step S2, the compound 2 and DSPE-mPEG 5000 The mass ratio is 1:(5~10).

7. The method for preparing a photothermal agent for treating allergic rhinitis according to claim 1, characterized in that: In the step S2, the temperature for evaporating and removing tetrahydrofuran under a nitrogen atmosphere is 22 to 25° C. and the time is 18 to 24 hours.

8. The method for preparing a photothermal agent for treating allergic rhinitis according to claim 1, characterized in that: In the step S2, the pore size of the filter membrane used for filtration is 0.2-0.45 μm.

9. A photothermal agent for treating allergic rhinitis, characterized in that: Prepared by the method described in any one of claims 1 to 8; the average size of the photothermal agent is 100 to 160 nm, and the molar absorption coefficient of the photothermal agent in the NIR-II region is 4000 to 4300 nm -1 cm -1 .

10. Use of the photothermal agent for treating allergic rhinitis as claimed in claim 9 in the preparation of a drug for treating allergic rhinitis.

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