A photothermal agent for treating allergic rhinitis and use thereof in the preparation of a medicament for allergic rhinitis

Abstract

The application discloses a photothermal agent for treating allergic rhinitis and application of the photothermal agent in preparation of allergic rhinitis drugs. 5000 The compound 2 is prepared by reacting the compound 1 with diethylene glycol-2-bromoethyl methyl ether, then the prepared compound 2 and DSPE-mPEG 5000 are dissolved in tetrahydrofuran to perform self-assembly and form nanoparticles, the nanoparticles have a red shift of an absorption wavelength due to a strong electron-withdrawing effect of the pyridine salt group, promote intramolecular charge transfer, have deeper tissue penetration ability, can effectively reach a deep part of a nasal mucosa and treat deep inflammation; the RH@NPs prepared by the application have high efficient photothermal conversion performance and sensitivity of photodynamic therapy, so that PTT and PDT can be used in synergistic treatment of allergic rhinitis, and the treatment effect is greatly improved, and a new way for treating allergic rhinitis is opened up.

Description

TECHNICAL FIELD

[0001] The application 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 preparation of a medicine for allergic rhinitis. BACKGROUND

[0002] Allergic rhinitis is one of the most common chronic diseases in the world, with a global prevalence rate of up to 20% to 30%, and has shown an upward trend in recent years. Allergic rhinitis is an IgE-mediated allergic reaction of the nasal cavity caused by inhaled allergens, and is manifested as sneezing, nasal congestion, nasal itching and rhinorrhea. In clinical practice, drug therapy is the most commonly used treatment method, but the treatment effect varies greatly among individuals. According to statistics, as high as 29% of children and 62% of adults only partially relieve or do not relieve symptoms after receiving single drug therapy, and long-term drug use 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 treatment methods that can target the local area to relieve local symptoms without causing systemic adverse reactions.

[0003] Phototherapy, as a new, safe and non-invasive treatment method, shows broad application prospects in the treatment of diseases through photothermal effect (PTT) and photodynamic effect (PDT). PTT has the advantages of high efficiency, controllability and low toxicity, and mild PTT can play a role in regulating immunity and regulating epithelial-mesenchymal transition (EMT) in the inflammatory state. PDT, as an effective photodynamic process, can activate the production of reactive oxygen species (ROS) through light irradiation. Reactive oxygen can act as a cell signaling molecule to maintain redox balance and promote health. Based on this property, PDT has been studied for the treatment of other diseases, but there are few reports on the treatment of allergic rhinitis using PTT and PDT in combination. In addition, current photothermal agents mainly focus on using light in the ultraviolet / visible spectrum for phototherapy, ignoring the use of near-infrared light (NIR), especially NIR-II region light with a wavelength range of 1000-1700nm, which has potential effectiveness in the treatment of rhinitis. Moreover, the amount of ROS generated by the photothermal agent in the prior art is uncontrollable, but a high content of ROS can have a cytotoxic effect. At the same time, due to the obstruction of nasal bone and soft tissue, the material properties need to be adjusted to select the light power to achieve the desired penetration depth while maintaining good biological safety during treatment. SUMMARY

[0004] In view of the problems in the prior art, the present application provides a photothermal agent for treating allergic rhinitis and an application thereof in the preparation of allergic rhinitis drugs, so as to solve the technical problems that the single PTT method cannot effectively achieve sufficient treatment of allergic rhinitis in the prior art, and the existing photothermal agent cannot effectively utilize NIR-II region light during the PDT treatment process, and the amount of ROS generated is uncontrollable, and the effect of the PDT therapy in the treatment of allergic rhinitis cannot be fully played.

[0005] The present application is realized by the following technical solutions:

[0006] A preparation method of a photothermal agent for treating allergic rhinitis, comprising the following steps:

[0007] S1: under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether is added to a chloroform solution of compound 1, and after stirring reaction, concentration and purification are performed to obtain compound 2;

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

[0009]

[0010] S2: the compound 2 and DSPE-mPEG 5000 are dissolved in tetrahydrofuran, ultrasonically mixed uniformly, then the mixed solution is injected into water to make the system uniformly dispersed, then tetrahydrofuran is evaporated under a nitrogen atmosphere, and after filtration, the RH@NPs-based NIR-II region photothermal agent is obtained.

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

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

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

[0014] Preferably, in the step S1, a chromatographic column purification method is used for purification treatment, and in the chromatographic 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 the step S2, the mass ratio of the compound 2 to DSPE-mPEG 5000 is 1:(5-10).

[0016] Preferably, in the step S2, the temperature for evaporating and removing the tetrahydrofuran under the nitrogen atmosphere is 22-25 DEG C, and the time is 18-24h.

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

[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 100-160 nm, the molar absorption coefficient of the photothermal agent in the NIR-II region is 4000-4300 M -1 cm -1 ).

[0019] The above-mentioned photothermal agent for treating allergic rhinitis is applied to the preparation of a drug for allergic rhinitis.

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

[0021] The application discloses a preparation method of a photothermal agent for treating allergic rhinitis. The photothermal agent prepared by the method has high efficient photothermal conversion performance and sensitivity of photodynamic therapy, so that photothermal therapy (PTT) and photodynamic therapy (PDT) can be combined to treat allergic rhinitis, the treatment effect is greatly improved, and a new treatment method for allergic rhinitis is provided. The PTT uses the photothermal agent to absorb light energy of a specific wavelength and convert the light energy into heat energy, mildly increases local temperature, helps to relieve swelling and congestion of nasal mucosa, and reduces allergic reaction. The PDT uses active oxygen species generated by a photosensitizer under light to mobilize the antioxidant system of mucosa, adjust immune response, and reduce release of inflammatory mediators. The combination of the two therapies not only improves the nasal environment from a physical aspect, but also adjusts immune response of the body from a root cause, and provides a more comprehensive, efficient and less side-effect treatment scheme for allergic rhinitis patients. In addition, in the preparation process of the photothermal agent, compound 1 with two pyridine groups is reacted with diethylene glycol-2-bromoethyl methyl ether to prepare ether oxygen chain substituted compound 2. In the process, the pyridine salt group acts as a strong electron-attracting group, enhances the push-pull electron effect, promotes intramolecular charge transfer, causes red shift of the absorption wavelength, improves the penetration depth of rhinitis treatment, and reduces tissue light damage. In addition, the ether oxygen chain is used to substitute the pyridine group, the electron-attracting ability is enhanced, the self-assembly ability is maintained, the coupling between excited states is promoted, and the photodynamic behavior is beneficial. Meanwhile, in the substitution process of the ether oxygen chain, diethylene glycol-2-bromoethyl methyl ether is added into a chloroform solution of compound 1 by controlling the dropping sequence, unilateral pyridine substitution of compound 1 is realized, the symmetric C2v point group D-A structure of compound 1 is changed into the asymmetric C1 point group D-A RH structure, the molecular polarity and dipole moment are increased, the self-assembly of compound 2 is promoted, the cross ability between systems is enhanced, and efficient photodynamic process is realized. Then, the compound 2 and DSPE-mPEG 5000Compound 2 is self-assembled into nanoparticles in tetrahydrofuran, the self-assembly process makes the intermolecular interaction more close, which is helpful for the energy transfer and coupling between the excited state molecules, because the interaction between the excited state molecules is the key to produce the photodynamic effect in the photodynamic process, through self-assembly, more excited state molecules can be in close spatial position, thereby increasing the opportunity of their interaction and improving the generation efficiency of photodynamic effect; the nanoparticles formed by self-assembly can better respond to long-wavelength light with high tissue penetration ability due to the strong electron-withdrawing effect of the pyridine salt group, which promotes intramolecular charge transfer and causes red shift of the absorption wavelength, so as to treat nasal mucosa inflammation by penetrating through the nasal bone and soft tissue; at the same time, the nanoparticles have moderate photothermal conversion efficiency due to the substitution of ether oxygen chain, which can rapidly warm up to the ideal treatment temperature under laser irradiation, while maintaining good thermal stability, and produce good PTT effect; in addition, the nanoparticles can produce superoxide free radicals (·O2 - ) under light conditions, and the near-infrared two-zone excited state has a short lifetime, so the generated free radicals are at a relatively low level, which can be used as a cell signaling molecule under low dose to trigger antioxidant defense, promote health, avoid cell damage caused by excessive treatment, and achieve good PDT effect; and in the in vitro experiment, even at a higher concentration, the survival rate of the nasopharyngeal epithelial cells is not greatly affected by the RH@NPs, which shows good biocompatibility, and the RH@NPs can be rapidly enriched in the nasal cavity and rapidly removed after laser irradiation, reducing the side effects caused by long-term retention. The preparation method of the present application is simple, reasonable in design, and the prepared RH@NPs have good PTT and PDT dual effects, and also have good penetration depth, which effectively improves the long-term health improvement of the nasal mucosa.

[0022] Further, in the step S1, the reaction temperature is 60-90 DEG C during the temperature rising reaction, and the reaction time is 12-24 h, and the appropriate temperature can improve the reaction rate, so that the reactants can be more fully contacted and reacted, and in the temperature range of 60-90 DEG C, the reaction rate is moderate, which can ensure the reaction and avoid unnecessary side reactions or degradation of the product caused by high temperature, and sufficient reaction time allows the reactants to fully react, thereby improving the yield and purity of the product.

[0023] Further, in the step S1, the molar ratio of the compound 1 to diethylene glycol-2-bromoethyl methyl ether is 1:(1-2), and the appropriate molar ratio can ensure sufficient contact and reaction between the reactants, thereby improving the reaction efficiency. When the molar ratio of the compound 1 to diethylene glycol-2-bromoethyl methyl ether is controlled in the range of 1:(1-2), it can be ensured that there is enough diethylene glycol-2-bromoethyl methyl ether to react with the 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] Further, in the step S1, the vacuum concentration method is used for concentration treatment, and in the vacuum concentration process, the vacuum degree is-0.05 to-0.1 MPa, which can make the solvent contained in RH@NPs completely evaporated, and RH@NPs with high purity can be obtained.

[0025] Further, in the step S1, the chromatography column purification method is used for purification treatment, and in the chromatography column purification process, the stationary phase is silica gel, and the mobile phase is a mixture of methanol and dichloromethane with a volume ratio of 1:25, which can shorten the purification time and efficiently obtain RH@NPs photothermal agent with high purity.

[0026] Further, in the step S2, the mass ratio of the compound 2 to DSPE-mPEG 5000 The mass ratio of the compound 2 to DSPE-mPEG

[0027] Further, in the step S2, the temperature for evaporating and removing tetrahydrofuran under a nitrogen atmosphere is 22-25℃, and the time is 18-24h, which can obtain stable RH@NPs.

[0028] Further, in the step S2, the pore size of the filter membrane used for filtration is 0.2-0.45μm, which can exclude large particle size nanometers that are not suitable for biological applications. BRIEF DESCRIPTION OF DRAWINGS

[0029] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following will briefly introduce the drawings needed to be used in the embodiments. It should be understood that the following drawings only show some embodiments of the present application, and therefore should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0030] Figure 1Synthesis procedure and photophysical property test results of photothermal agent RH@NPs prepared in Example 1 of the present application, wherein: (a) is the self-assembly procedure of RH@NPs; (b) is the comparison of 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 = 808 nm); (d) is the absorption spectra 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 = 808 nm); (g) is the absorption spectra of RH@NPs (ε: molar absorption coefficient); (h) is the photothermal heating curve of RH@NPs;

[0031] Figure 2 Radical capture test of photothermal agent RH@NPs prepared in Example 1 of the present application;

[0032] Figure 3 In vitro cell experiment of photothermal agent RH@NPs prepared in the present application: wherein, (a) is the CCK8 experiment to evaluate the cell viability of NP69 under different concentrations (0, 0.5, 1, 2 mg mL -1 ) of RH@NPs; (b) is the fluorescence microscope detection of the uptake of RH@NPs by human nasopharyngeal epithelial NP69 cells; (c) is the fluorescence quantification chart of Fig. (b)

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

[0034] Figure 5 Photothermal treatment of photothermal agent RH@NPs prepared in the present application on allergic rhinitis in mice: wherein, (a) is the flow chart of in vivo experiment; (b) is the general behavior evaluation of allergic rhinitis mice; (c) is the content of serum histamine and allergen-specific IgE; (d) is the pathological staining result of nasal mucosa tissue section of allergic rhinitis mice;

[0035] Figure 6 Therapeutic effect of photothermal agent RH@NPs prepared in the present application on oxidative stress and inflammation of allergic rhinitis mice: wherein, (a) is the level of inflammatory factors in the nasal mucosa; (b) is the level of glutathione synthesis metabolism related factors in the nasal mucosa; (c) is the mRNA level of EMT related factors in the nasal mucosa. DETAILED DESCRIPTION

[0036] To enable persons skilled in the art to understand the features and effects of the present application, the following is a general description and definition of the terms and phrases mentioned in the specification and claims. Unless otherwise specified, all technical and scientific words used herein are understood to have the usual meaning as understood by those skilled in the art of the present application, and in the event of a conflict, the definition in the specification shall prevail.

[0037] Theories and mechanisms described and disclosed herein, whether correct or not, should not be considered limiting on the scope of the present application, that is, the present application can be practiced without regard to any particular theory or mechanism.

[0038] Herein, all features defined in the form of numerical ranges or percentage ranges such as values, amounts, contents and concentrations are for the sake of brevity and convenience. Accordingly, the description of numerical ranges or percentage ranges should be considered to have encompassed and specifically disclosed all possible sub-ranges and individual numerical values within the range (including integers and fractions).

[0039] Herein, unless otherwise specified, "comprise", "include", "contain", "have" or similar words encompass the meaning of "consist of" and "consist essentially of", for example, "A comprises a" encompasses the meaning of "A comprises a and other" and "A comprises only a".

[0040] Herein, for the sake of brevity, all possible combinations of the technical features in each embodiment or example are not described. Therefore, as long as the combinations of the technical features do not contradict each other, the technical features in each embodiment or example can be combined arbitrarily, and all possible combinations should be considered to be within the scope of the present specification.

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

[0042] S1: Under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether was added to a chloroform solution of compound 1, wherein the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether was 1:(1-2). After stirring until homogeneous, the system was heated to 60-90°C and reacted for 12-24 hours. Concentration was performed under vacuum at -0.05 to -0.1 MPa, followed by purification by column chromatography. During column chromatography, silica gel was used as the stationary phase, and a mixture of methanol and dichloromethane (volume ratio 1:25) was used as the mobile phase. After purification, compound 2 was obtained. In this step, diethylene glycol-2-bromoethyl methyl ether was added in three batches, one-third of the calculated amount each time, stirred for 30 minutes, before adding the next batch.

[0043] The structural formula of compound 1 is:

[0044]

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

[0046]

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

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

[0049] In the present application, 1-(2-Bromoethoxy)-2-(2methoxyethoxy)ethane is used to synthesize RH with pyridine salt structure, i.e. compound 2, by nucleophilic substitution reaction with the pyridine group in compound 1. In this process, the pyridine salt group is introduced as a strong electron-attracting group, which enhances the push-pull electron effect, promotes intramolecular charge transfer, causes red shift of the absorption wavelength, improves the penetration depth of rhinitis treatment, and reduces tissue photodamage. The use of ether oxygen chain instead of alkyl chain enhances the electron-attracting ability while maintaining the self-assembly ability, promoting the coupling between excited states and being conducive to photodynamic behavior. Unilateral pyridine substitution changes the symmetric C2v point group D-A structure of compound 1 to the asymmetric C1 point group D-ARH structure, increases the molecular polarity and dipole moment, promotes the self-assembly of RH, enhances the inter-system crossing ability, and realizes efficient photodynamic generation. In addition, the prepared RH@NPs exhibit photothermal conversion ability under 808 nm and 1064 nm laser irradiation, and 1064 nm laser is more suitable for treating deep rhinitis due to its deeper tissue penetration ability. RH@NPs have moderate photothermal conversion efficiency (34%), and can produce superoxide anion (·O2 - ), which is the main active oxygen species generated by photodynamic effect. At the same time, RH@NPs exhibit excellent in vitro biocompatibility, and even at a concentration as high as 2 mg / mL, the survival rate of nasopharyngeal epithelial cell line NP69 is not significantly affected. Through the nasal spray administration, RH@NPs can rapidly enrich in the nasal mucosa, and reach the ideal photothermal temperature after laser irradiation, and be effectively cleared within a few hours, reducing the side effects caused by systemic absorption. In addition, in the OVA-induced AR mouse model, by nasal spray administration of RH@NPs combined with laser irradiation, the acute symptoms of AR are significantly reduced, the allergic mediators such as histamine and OVA-specific IgE in serum are reduced, and the epithelial damage of nasal mucosa is improved.

[0050] The present application also discloses a photothermal agent for treating allergic rhinitis prepared by the above method, wherein the average size of the RH@NPs-based NIR-II region photothermal agent 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 application also explores the photothermal and photodynamic properties of RH@NPs under different concentrations and laser power densities, as well as the therapeutic effects in different animal models, providing multiple application scenarios and the possibility of technical adjustment. The technical solution of the application is innovative, providing a new type of safe, effective, and controllable treatment method for allergic rhinitis. Through precise molecular design and regulation of optical physical properties, RH@NPs can achieve high-efficiency PTT and PDT effects in the NIR-II region, providing a new strategy for the treatment of allergic rhinitis.

[0052] The application proposes a photothermal agent RH@NPs for the synergistic treatment of allergic rhinitis by photothermal effect (PTT) and photodynamic effect (PDT), which has the following advantages:

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

[0054] (2) High photothermal conversion efficiency: RH@NPs exhibit moderate photothermal conversion efficiency, which can rapidly warm up to the ideal treatment 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 - ), which can act as cell signaling molecules at low doses, triggering antioxidant defenses and promoting health, while controlling the amount of free radicals produced by controlling the irradiation time and power, and the concentration of free radicals produced by near-infrared II region materials is low, which can effectively avoid cell damage caused by excessive treatment.

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

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

[0058] (6) Comprehensive treatment 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 prospect: The RH@NPs of the present application provide a safe, effective and controllable new choice for the clinical treatment of allergic rhinitis, which is expected to reduce the economic burden of patients and improve the quality of life. The RH@NPs of the present application provide new ideas and methods for the application of phototherapy in other disease treatment fields, and promote scientific research and technical development in related fields.

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

[0061] The following examples use conventional apparatus in the art. The experimental methods in the following examples, unless otherwise specified, are usually carried out according to conventional conditions, or according to the conditions recommended by the manufacturer. Various raw materials are used in the following examples, unless otherwise specified, and conventional commercially available products are used, which are conventional specifications in the art. In the specification of the present application and in the following examples, unless otherwise specified, "%" means weight percent, "parts" means weight parts, and the ratio means weight ratio.

[0062] Example 1

[0063] A preparation method of a photothermal agent for treating allergic rhinitis, comprising the following steps:

[0064] S1: Under N2protection, 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 containing CHCl3(10 mL), stirred at 90°C for 24 h, the reaction was cooled to room temperature, and the solvent was concentrated under vacuum. Finally, purification by column chromatography (silica gel, MeOH / DCM = 1 / 25) gave RH blue-green solid (yield 20%), which is compound 2, represented as RH.

[0065] 1 H NMR (500 MHz, 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 (126 MHz, CDCI3) d / ppm 155.7, 150.4, 147.6, 145.5, 139.2, 133.5, 130.6, 130.2, 127.7, 127.0, 124.2, 121.6, 114.5, 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 the compound 2 and 5 mg of DSPE-mPEG 5000 were dissolved in 1 mL of tetrahydrofuran, and then the mixture was mixed uniformly by ultrasonic, and then the mixture was injected into 9 mL of deionized water to make the system uniformly dispersed, and then the system was placed in a nitrogen atmosphere, and the system was treated by stirring at 22°C for 18 h, and then tetrahydrofuran was evaporated, and then a filter membrane with a pore size of 0.45 μm was used for filtration, and then a photothermal agent for the NIR-II region based on RH@NPs was prepared.

[0067] The technical effects of the present application are illustrated by using the RH and RH@NPs prepared in this embodiment, and specifically:

[0068] Figure 1 For the synthesis process of the photothermal agent RH@NPs in the present application and the photophysical property test results of the RH and RH@NPs synthesized in Example 1, (a) is the self-assembly process of RH@NPs; (b) is the comparison of the absorption spectrum and the emission spectrum of compound 1 (SW6) and RH; (c) is the absorption spectrum and the emission spectrum of RH (in dimethyl sulfoxide (DMSO), concentration = 10 -5 mol / L, λ ex = 808 nm); (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 spectrum and the emission spectrum of RH@NPs (in H2O, concentration = 10 -5 mol / L, λ ex = 808 nm); (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 pyridine cyanide unit is introduced into RH molecule as a strong electron-withdrawing group, which enhances the push-pull electron effect and the electron transfer ability. Compared with compound 1, the red shift of the absorption wavelength of RH can improve the penetration depth of treatment, and the photothermal effect of RH@NPs can be used for photothermal therapy of tumors. Figure 1As shown in (b), (c), and (d), the absorption and emission peaks of RH molecules (998 nm) are closer to 1000 nm. Compared to RH molecules, the absorption spectrum of RH@NPs nanoparticles is significantly broader, indicating the coexistence of multiple aggregation states. Furthermore, after assembly into nanoparticles, their absorption spectrum exhibits a slight blue shift, suggesting a Coulomb coupling effect among the molecules in the coated nanoparticles. Moreover, during the material construction process, only a red shift to a longer wavelength band followed by a blue shift can guarantee the penetration depth of the RH@NPs nanoparticles. Figure 1 (e) shows that a uniformly shaped spherical nanoparticle with a size of 100 nm is formed; from Figure 1 (h) shows 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 this invention was specifically performed by using 2',7'-dichlorodihydroxybenzyl ol (DCFH) as a fluorescent probe, and adding RH@NPs photothermal agent and different reactive oxygen species scavengers for fluorescence quenching experiments. The reactive oxygen species scavengers were NaN3 (SA, used for...) 1 O2), Vitamin C (VC, used for O2) - ·) and isopropanol (IPA, used for ·OH), test results are shown in Figure 2 As shown, by Figure 2 It can be seen that the fluorescence intensity was significantly suppressed only after the addition of VC, indicating that RH@NPs produce O2 under both 808nm and 1064nm illumination. - It has type I photodynamic therapy capabilities.

[0070] Figure 3 In vitro cell experiments of the photothermal agent RH@NPs prepared in this invention: (a) is a CCK8 assay to evaluate different concentrations of RH@NPs (0, 0.5, 1, 2 mg / mL). -1 (a) shows the viability of NP69 cells; (b) shows the uptake of RH@NPs by human nasopharyngeal epithelial NP69 cells by fluorescence microscopy; (c) shows the quantitative fluorescence graph of (b). Figure 3 (a) It can be seen that RH@NPs have good biocompatibility, do not affect cell proliferation, and are non-cytotoxic; Figure 3 As shown in (b) to (c), RH@NPs can be effectively taken up by nasopharyngeal epithelial cells and rapidly 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 application in the nasal mucosa of mice are as follows: the enrichment, maintenance and clearance of RH@NPs in the nasal mucosa of mice are evaluated by nasal spray administration, and the change in nasal temperature after laser irradiation is monitored by infrared thermal imaging technology. As can be seen from the figure, compared with the blank control (PBS group), the nasal temperature of the RH@NPs group rapidly rises to 39.1℃ within 4 min and is maintained for at least 20 min, and no obvious nasal mucosa swelling occurs, indicating that the photothermal effect mediated by RH@NPs has the characteristics of rapidness, stability and moderation. The highest temperature in the nasal cavity decreases within 3 hours and returns to normal within about 6 hours. This temperature change indicates that RH@NPs are rapidly cleared from the nasal mucosa through nasal cilia 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 application is as follows: (a) is the flow chart of in vivo experiment; (b) is the general behavior evaluation of allergic rhinitis mice, in which the left figure is the sneezing frequency data of the disease mice, and the right figure is the frequency data of the disease mice scratching the nose; (c) is the evaluation of the level of allergic mediators in the serum of allergic rhinitis mice, in which the left figure is the change in the content of histamine in the blood of the disease mice, and the right figure is the change in the content of allergen-specific IgE in the blood of the disease mice; (d) is the pathological staining result of the nasal mucosa tissue section of allergic rhinitis mice, in which the epithelial cell thickness of the respiratory area of the nasal septum is quantitatively detected by HE staining to evaluate the damage of the nasal mucosa, and the hyperplasia of the goblet cells of the nasal mucosa is evaluated by PAS staining. As can be seen from the figure, the treatment of RH@NPs given by nasal spray has a good therapeutic effect on allergic rhinitis mice, which is manifested in the reduction of allergic symptoms such as sneezing and scratching of the nose of the disease mice, the reduction of histamine and allergen-specific IgE in the blood, the reduction of swelling damage of the nasal mucosa of the nasal septum, and the reduction of goblet cell hyperplasia.

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

[0074] Example 2

[0075] A preparation method of a photothermal agent for treating allergic rhinitis, comprising the following steps:

[0076] S1: under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether is added to a chloroform solution of compound 1 in three batches, after each addition of diethylene glycol-2-bromoethyl methyl ether, stirring for 30 min, then 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 is 1:1, after uniform stirring, the system is heated to 60°C and reacted for 24 h, under a vacuum degree of -0.05 MPa, concentrated by vacuum concentration, 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, after completion of the purification process, compound 2 is prepared.

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

[0078]

[0079] S2: compound 2 and phospholipid methoxy polyethylene glycol (DSPE-mPEG 5000 ) are dissolved in tetrahydrofuran at a mass ratio of 1:5, ultrasonically mixed uniformly, then the mixed solution is injected into water, the system is uniformly dispersed under a cell crusher, then the system is placed in a nitrogen atmosphere and stirred at 22°C for 24 h, tetrahydrofuran is evaporated off, and a photothermal agent for the NIR-II region based on RH@NPs is prepared after filtration with a filter membrane with a pore size of 0.2 μm.

[0080] Example 3

[0081] A preparation method of a photothermal agent for treating allergic rhinitis, comprising the following steps:

[0082] S1: under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether is added to a chloroform solution of compound 1 in three batches, after each addition of diethylene glycol-2-bromoethyl methyl ether, stirring for 30 min, then 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 is 1:2, after uniform stirring, the system is heated to 90°C and reacted for 12 h, under a vacuum degree of -0.1 MPa, concentrated by vacuum concentration, 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, after completion of the purification process, compound 2 is prepared.

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

[0084]

[0085] S2: the compound 2 and phospholipid methoxy polyethylene glycol (DSPE-mPEG 5000 ) are dissolved in tetrahydrofuran at a mass ratio of 1: (5-10), ultrasonic mixing is uniform, then the mixed solution is injected into water, the system is uniformly dispersed under a cell crusher, then the system is placed in a nitrogen atmosphere, the system is stirred at 25 DEG C for 18h, tetrahydrofuran is evaporated, after filtration by using a filter membrane with a pore size of 0.45 mu m, a photothermal agent for the NIR-II region based on RH@NPs is prepared.

[0086] Example 4

[0087] A preparation method of a photothermal agent for treating allergic rhinitis, comprising the following steps:

[0088] S1: under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether is added to a chloroform solution of compound 1 in three batches, after adding diethylene glycol-2-bromoethyl methyl ether each time, stirring is carried out for 30 min, then the next batch of diethylene glycol-2-bromoethyl methyl ether is added, the molar ratio of compound 1 to diethylene glycol-2-bromoethyl methyl ether is 1:1.5, after uniform stirring, the system is heated to react at 80 DEG C for 20h, under a vacuum degree of-0.1 MPa, concentration treatment is carried out by using a vacuum concentration method, and purification is carried out by using a chromatographic column purification method, in the chromatographic column purification process, silica gel is used as a stationary phase, and a mixture of methanol and dichloromethane at a volume ratio of 1:25 is used as a mobile phase, after completion of the purification treatment, compound 2 is prepared.

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

[0090]

[0091] S2: the compound 2 and phospholipid methoxy polyethylene glycol (DSPE-mPEG 5000 ) are dissolved in tetrahydrofuran at a mass ratio of 1:8, ultrasonic mixing is uniform, then the mixed solution is injected into water, the system is uniformly dispersed under a cell crusher, then the system is placed in a nitrogen atmosphere, the system is stirred at 23 DEG C for 20h, tetrahydrofuran is evaporated, after filtration by using a filter membrane with a pore size of 0.3 mu m, a photothermal agent for the NIR-II region based on RH@NPs is prepared.

[0092] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application and not to limit the protection scope of the present application, although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced equivalently without departing from the essence and scope of the technical solutions of the present application.

Claims

1. A method for preparing a photothermal agent for treating allergic rhinitis, characterized by, The method comprises the following steps: S1: under a nitrogen atmosphere, diethylene glycol-2-bromoethyl methyl ether is added into a chloroform solution of compound 1, after stirring reaction, compound 2 is prepared through concentration and purification; The structural formula of the compound 1 is: ； The structural formula of the compound 2 is: ； S2: Combine compound 2 and DSPE-mPEG 5000 Dissolve the mixture in tetrahydrofuran, mix it evenly with ultrasound, then inject the mixture into water to disperse the system evenly. Then evaporate the tetrahydrofuran under a nitrogen atmosphere, filter it, and obtain the photothermal agent for treating allergic rhinitis. In the step S1, the molar ratio of the compound 1 to diethylene glycol-2-bromoethyl methyl ether is 1:(1-2); In the step S2, the compound 2 is reacted with DSPE-mPEG 5000 in a mass ratio of 1:(5-10).

2. The method of claim 1, wherein the photothermal agent is prepared by the steps of: (a) dissolving the photothermal agent in a solvent; (b) adding a surfactant to the solution; (c) adding a polymer to the solution; and (d) adding a stabilizer to the solution. In the step S1, during the reaction, the reaction temperature is 60-90 ℃, and the reaction time is 12-24 h.

3. The method of claim 1, wherein the photothermal agent is prepared by the steps of: (a) dissolving the photothermal agent in a solvent; (b) adding a surfactant to the solution; (c) adding a polymer to the solution; and (d) adding a stabilizer to the solution. In the step S1, the concentration treatment is performed by using a vacuum concentration method, and in the vacuum concentration treatment process, the vacuum degree is-0.05--0.1 MPa.

4. The method of claim 1, wherein the photothermal agent is prepared by the steps of: (a) dissolving the photothermal agent in a solvent; (b) adding a surfactant to the solution; (c) adding a polymer to the solution; and (d) adding a stabilizer to the solution. In the step S1, the purification treatment is performed by using a chromatographic column purification method, and in the chromatographic column purification process, the stationary phase is silica gel, and the mobile phase is a mixture of methanol and dichloromethane with a volume ratio of 1:

25.

5. The method of claim 1, wherein the photothermal agent is prepared by the steps of: (a) dissolving the photothermal agent in a solvent; (b) adding a surfactant to the solution; (c) adding a polymer to the solution; and (d) adding a stabilizer to the solution. In the step S2, the temperature for evaporating and removing tetrahydrofuran under a nitrogen atmosphere is 22-25 ℃, and the time is 18-24 h.

6. A 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 in the filtration is 0.2-0.45 μm.

7. A photothermal agent for treating allergic rhinitis, characterized by, made by the method according to any one of claims 1-6; the average size of the photothermal agent 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 .

8. Use of a photothermal agent for treating allergic rhinitis according to claim 7 in the preparation of a medicament for allergic rhinitis.

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