External traditional Chinese medicine composition for treating allergic rhinitis as well as preparation method and application of external traditional Chinese medicine composition

Abstract

The invention provides an external traditional Chinese medicine composition for treating allergic rhinitis. The external traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 160 to 240 parts of paniculate swallowwort root, 36 to 54 parts of cicada slough, 0.24 to 0.36 part of borneol and 0.24 to 0.36 part of antrodia camphorata. The invention further provides a preparation method and application of the external traditional Chinese medicine composition. Pharmacodynamic tests prove that the medicinal effect of the traditional Chinese medicine composition is obviously superior to that of a prescription disclosed in the application number 201010238755.7, and the traditional Chinese medicine composition is more favorable for being prepared into a gel and exerts a better medicinal effect. The prepared external nasal drop can remarkably improve clinical symptoms of allergic rhinitis and is high in safety, meanwhile, the invention further provides a method for preparing the external nasal drop, and the method is simple in process and easy to produce.

Description

Technical Field

[0001] This invention relates to a topical traditional Chinese medicine composition for treating allergic rhinitis, belonging to the field of traditional Chinese medicine. Background Technology

[0002] Allergic rhinitis (AR) is the most common chronic inflammatory disease of the nasal mucosa in children. The prevention and treatment of AR has become a global health issue. The key to its treatment lies in avoiding allergens and using medication, but satisfactory results have not yet been achieved. Traditional Chinese medicine has significant advantages in treating AR; therefore, researching and developing safe and effective topical nasal drops for AR treatment is of great importance. Nasal sprays are a clinically effective method, but current spray formulations mostly use water as a solvent. After being sprayed into the nasal cavity, gravity and other factors make it difficult for the spray to remain at the site of action for an extended period, thus reducing clinical efficacy.

[0003] Application No. 201010238755.7 discloses a pharmaceutical composition for treating allergic rhinitis, its preparation method, and its uses. The patent provides a pharmaceutical composition for treating allergic rhinitis, which is a preparation made from the following traditional Chinese medicine raw materials in the indicated weight ratios: 14-26 parts of *Cynanchum paniculatum*, 3.15-5.85 parts of *Cicada slough*, 0.021-0.039 parts of borneol, and 0.21-0.39 parts of bezoar. Among the raw materials disclosed in this patent, artificial bezoar has the functions of clearing heat and detoxifying, and refreshing the mind. Studies have shown that artificial bezoar has a significant inhibitory effect on all stages of inflammation and also has antioxidant effects. However, artificial bezoar is not suitable for preparation into a gel dosage form.

[0004] Please provide any shortcomings compared to this application. Summary of the Invention

[0005] This invention provides a topical traditional Chinese medicine composition for treating allergic rhinitis, its preparation method, and its uses.

[0006] This invention provides a topical traditional Chinese medicine composition for treating allergic rhinitis, which is prepared from the following raw materials in the indicated weight ratios:

[0007] Xu Changqing 160-240 parts, Cicadae Periostracum 36-54 parts, Borneol 0.24-0.36 parts, Antrodia camphorata 0.24-0.36 parts.

[0008] Preferably, it is prepared from the following raw materials in the indicated weight ratios:

[0009] 200 parts of Xu Changqing, 45 parts of cicada molting, 0.3 parts of borneol, and 0.3 parts of Antrodia camphorata.

[0010] The traditional Chinese medicine composition of this invention is prepared as a commonly used external preparation by adding pharmaceutically acceptable excipients or auxiliary ingredients, with paeonol extracted from Xu Changqing, water extracts of cicada molting and Antrodia camphorata, and borneol as active ingredients.

[0011] The aforementioned topical preparations include nasal drops, nasal washes, nasal sprays, nasal aerosols, nasal ointments, nasal creams, and nasal gels.

[0012] The nasal gel is prepared by extracting paeonol from Cynanchum paniculatum, decocting it with cicada molting and Antrodia camphorata to obtain a medicinal solution, preparing borneol and paeonol into nanoparticles, adding propylene glycol, mixing the medicinal solution, and a gel matrix.

[0013] The nasal gel contains the following raw materials and matrix per 100ml:

[0014] 1.2g of nanoparticles, 5g of propylene glycol, 70ml of drug solution, plus 2g of poloxamer P188, 2g of polyethylene glycol PEG6000, 0.4g of potassium sorbate, and 18g of poloxamer P407;

[0015] The 70ml of liquid contains 4.5g of raw cicada molting and 0.03g of Antrodia camphorata.

[0016] The present invention also provides a method for preparing the aforementioned topical traditional Chinese medicine composition for treating allergic rhinitis, which includes the following steps:

[0017] a. Preparation of the medicinal solution: Weigh the raw materials according to the weight ratio; extract the paeonol distillate by steam distillation of Cynanchum paniculatum, cool, settle, and filter to obtain crystals for later use; mix the extracted Cynanchum paniculatum residue with Cicadae Periostracum and Antrodia camphorata and decoct in water to extract, then mix the decoction with the remaining medicinal solution after steam distillation, filter and concentrate, centrifuge the concentrate, take the supernatant after concentration, add 1% chitosan solution, let stand and centrifuge, take the supernatant after centrifugation, concentrate the supernatant to obtain the medicinal solution;

[0018] b. Preparation of nanoparticles: Borneol and paeonol were dissolved in PEG400 to form a borneol-paeonol solution. The solution was slowly added dropwise to a 2% chitosan acetate solution under magnetic stirring. The pH was adjusted to 3.0 with NaOH solution. Then, TPP solution was slowly added dropwise under magnetic stirring. The solution was stirred continuously to obtain borneol-paeonol inclusion complex nanoparticles.

[0019] c. Preparation of gel: Take the nanoparticles prepared in step b, add propylene glycol and drug solution and mix well, then add poloxamer P188, polyethylene glycol PEG6000, potassium sorbate and poloxamer P407, disperse them evenly, store at 4-10℃ overnight to obtain gel.

[0020] In step b, the mass ratio of chitosan to paeonol is 6-8:1, preferably 6:1; the stirring speed is 1000 r; and the mass concentration ratio of chitosan to sodium tripolyphosphate is 6:1.

[0021] The present invention also provides the use of the aforementioned traditional Chinese medicine composition in the preparation of nasal medicines for treating allergic rhinitis.

[0022] The aforementioned medication is used to treat allergic rhinitis caused by latent heat in the lung meridian.

[0023] The inventors removed artificial bezoar from the formula disclosed in patent application number 201010238755.7. Clinical application revealed that its anti-inflammatory effect was not ideal for patients with recurrent allergic rhinitis. The prescription needs to be further optimized and improved in order to further enhance clinical efficacy.

[0024] Antrodia cinnamomea has been used as a traditional Chinese medicine in Taiwan for over two hundred years. It is a unique and rare medicinal fungus, renowned as the "King of Herbs." According to traditional Chinese medicine classification, Antrodia cinnamomea is categorized as a heat-clearing and detoxifying herb. Its properties are pungent, bitter, and slightly sweet; it is a cold-natured herb; and it enters the liver, gallbladder, and lung meridians. Its functions include clearing heat and detoxifying, reducing swelling and dissipating nodules, relieving sore throat, clearing liver fire, and dispersing blood stasis and relieving pain.

[0025] Modern research results indicate that *Antrodia camphorata* belongs to the genus *Polyporus* of the family Polyporaceae. The fruiting body and mycelium of *Antrodia camphorata* contain various physiologically active substances, including active polysaccharides, terpenoids, maleic acid and succinic acid and their derivatives, as well as ubiquinone compounds. The polysaccharides in *Antrodia camphorata* mainly have a β-D-glucan structure, which can enhance the body's immune function by stimulating immune cells such as macrophages and lymphocytes. Simultaneously, *Antrodia camphorata* polysaccharides also possess numerous physiological activities such as antioxidation, anti-inflammation, antithrombosis, and antiglycation. The triterpenoids in *Antrodia camphorata* have multiple effects, including regulating immunity, antioxidation, anti-inflammation, and anti-allergy. In the formulation of this invention, *Cynanchum paniculatum* is pungent and warm in nature, dispelling wind, opening the orifices, removing dampness, and relieving itching; its main active ingredient is paeonol. *Cicada slough* is sweet and cold in nature, clearing the head and eyes, and effectively dispelling external wind and internal wind. Borneol is bitter, pungent, and slightly cold in nature, opening the orifices, refreshing the mind, and guiding the medicine upwards. Therefore, the whole formula works to dispel wind, clear heat, and open the nasal passages, and is suitable for allergic rhinitis (nasal congestion) with latent heat syndrome in the lung meridian.

[0026] The advantages of using nanogel dosage form in this invention are: (1) improving the bioavailability of drugs and reducing the dosage; (2) enhancing the targeting of drugs; (3) achieving sustained release and controlled release effects; (4) enhancing the efficacy of traditional Chinese medicine or presenting new efficacy, and broadening the indications of the original drug; (5) enriching the dosage form selection and administration route of traditional Chinese medicine.

[0027] Pharmacological tests have demonstrated that the herbal composition of this invention is significantly more effective than the prescription disclosed in application number 201010238755.7, and is more suitable for preparation into a gel, thus exhibiting better efficacy. The prepared topical nasal drops can significantly improve the clinical symptoms of allergic rhinitis and have high safety. Furthermore, this invention also provides a method for preparing this drug, which is simple and easy to produce. Attached Figure Description

[0028] Figure 1 The effect of different concentrations of P407 on viscosity η;

[0029] Figure 2 The effect of different concentrations of P188 on viscosity η;

[0030] Figure 3 The effect of different concentrations of PEG6000 on viscosity η;

[0031] Figure 4 The effect of different concentrations of 1,2- on the viscosity η of propylene glycol;

[0032] Figure 5 Effects of the drug on IL-4-induced TNF-α, IL-1β, and IL-6 mRNA expression in HNEpCs (Note: Compared with the Control group, ## P<0.01; compared with the IL-4 model group, * P<0.05, ** P<0.01; compared with group A, & P<0.05, && P<0.01; compared with group B, ΔP<0.05);

[0033] Figure 6 Effects of different doses of Antrodia camphorata combination groups on IL-4-induced expression of TNF-α, IL-1β, and IL-6 mRNA in HNEpCs (Note: Compared with the Control group, ##P<0.01; compared with the model IL-4 group, *P<0.05, **P<0.01);

[0034] Figure 7 Effects of drugs on IL-4-induced expression of iNOS and COX2 proteins in HNEpCs (Note: Compared with the Control group, ##P<0.01; compared with the model IL-4 group, **P<0.01; compared with group A,

[0035] & P<0.05; compared with group B, Δ P<0.05). Detailed Implementation

[0036] Example 1: Preparation of the topical Chinese medicine nasal drops of the present invention

[0037] The pharmaceutical composition of the present invention is formulated as follows: 200g of Cynanchum paniculatum, 45g of Cicadae periostracum, 0.3g of Borneol, and 0.3g of Antrodia camphorata.

[0038] The preparation method of the present invention includes the following steps:

[0039] (1) Preparation of the medicinal solution: Mix the above-mentioned raw materials in a certain proportion. For example, extract paeonol using steam distillation of Cynanchum paniculatum, mix the residue with cicada slough and Antrodia camphorata, and extract using water extraction (1:8, twice) for 1 hour. Filter, combine the filtrates, concentrate to 1000 ml, centrifuge, take the supernatant, add 1% chitosan solution, let stand, centrifuge, take the supernatant, concentrate to 700 ml, and obtain the medicinal solution.

[0040] (2) Preparation of nanoparticles: Take 3 mg of borneol and 26 mg of paeonol, add 10 mL of PEG400 to dissolve them into a borneol-paeonol solution, and slowly add it dropwise to 25 mL of a chitosan solution containing 2% acetic acid (2 mg / mL) under magnetic stirring. Continue stirring for 10 min, adjust the pH to 5.0 with 1 mol / L NaOH solution, and then slowly add 10 mL of 1 mg / mL TPP solution at a rate of 2 drops / s under magnetic stirring. Continue stirring for 20 min to obtain borneol-paeonol inclusion complex nanoparticles.

[0041] (3) Preparation of gel: 1.2g of nanoparticles were mixed with 5g of propylene glycol and 70ml of drug solution. Then, 2g of poloxamer P188, 2g of polyethylene glycol PEG6000, 0.4g of potassium sorbate, and 18g of poloxamer P407 were added and dispersed evenly to make 100ml. The mixture was stored at 4-10℃ overnight to obtain the thermosensitive gel of the present invention.

[0042] Example 2: Selection of the matrix and screening of process parameters for the topical Chinese medicine nasal drop gel of the present invention.

[0043] 1. Evaluation Indicators for Thermosensitive Gels

[0044] Thermosensitive gels exhibit a unique "solution-gel-solution" state transition phenomenon, thus forming two temperature transition points: the gelling temperature (T1) and the melting temperature (T2). Between T1 and T2, the formulation exists in a gel state, and this temperature range is defined as the gelation range. The thermodynamic changes of the nasal irritation thermosensitive gel can be evaluated by measuring T1 and T2.

[0045] During temperature changes, the formulation undergoes a phase change, and its viscosity (η) also changes with increasing temperature. At lower temperatures, the formulation is liquid with low viscosity; as the temperature gradually increases, a phase change occurs, transforming it from a liquid to a semi-solid state, resulting in a significant viscosity change near the gelation temperature T1; with continued temperature increases until the melting temperature T2 is reached, the gel transforms into a liquid state, and the viscosity decreases. Therefore, the rheological changes of the nasal temperature-sensitive gel can be evaluated by measuring its viscosity.

[0046] 2. Experiment on selection of gelling agent matrix and screening of process parameters

[0047] 2.1 Investigation of Poloxamer P407

[0048] In addition to the dosage form matrix, the gel formulation also contains the drug solution, paeonol-borneol nanoparticles, 0.2% potassium sorbate as a preservative, and 1,2-propanediol as a suspending agent. Therefore, it is necessary to investigate the effects of these factors on the P407 blank thermosensitive gel.

[0049] P407 was added to deionized water at concentrations of 16%, 18%, 20%, 22%, and 24%. Paeonol-borneol nanoparticles were added to aqueous solutions of P407 at different concentrations. P407 was also added to the drug solution at concentrations of 16%, 18%, 20%, 22%, and 24%. The effects of the drug solution and inclusion complex on the gelation temperature were investigated. The results are shown in Table 1.

[0050] Table 1. Effects of drug solution and paeonol-borneol nanoparticles on T1 and T2 of P407 solution.

[0051]

[0052] P407 was added to deionized water at concentrations of 16%, 18%, 20%, 22%, and 24%. Potassium sorbate was added to aqueous solutions of P407 at different concentrations. Propylene glycol was also added to aqueous solutions of P407 at different concentrations to investigate the effects of potassium sorbate and propylene glycol on the gelation temperature. The results are shown in Table 2.

[0053] Table 2 Effects of potassium sorbate and 1,2-propanediol on T1 and T2 of P407 solution.

[0054]

[0055] The drug solution and paeonol-borneol nanoparticles are essential components of the formulation. Tables 1 and 2 show that using the drug solution to prepare P407 thermosensitive gel has a smaller impact on the gelation temperature compared to using water, and the overall temperature trend remains unchanged. Adding paeonol-borneol nanoparticles to the P407 blank gel also has no significant effect on the gelation temperature. Potassium sorbate, as a preservative, has virtually no effect on the gelation and melting temperatures. Therefore, it can be concluded that the drug solution, inclusion complex, and potassium sorbate have essentially no effect on the gelation temperature, and they should be added in future studies.

[0056] The thermosensitive gelation of poloxamer 407 is a result of the interaction between the hydrophilic PEO block and the hydrophobic PPO block, with the PPO block typically causing a decrease in gelation temperature. Therefore, polyethylene glycol and poloxamer 188 (hereinafter referred to as P188), which has a higher PEO block content, are often added to the formulation to obtain a suitable gelation temperature.

[0057] Thermosensitive gels were prepared using P407 as the matrix, P188 and PEG6000 as temperature regulators, and propylene glycol as a suspending agent. The effects of P407, P188, PEG6000, and propylene glycol on T1, T2, and η were mainly investigated.

[0058] The effect of P407 on T1, T2, and η

[0059] The prescribed amounts of paeonol-borneol nanoparticles, P188 2%, PEG6000 2%, and propylene glycol 5% were added to the drug solution, and P407 was added to prepare 16%, 18%, 20%, 22%, and 24% (w / w) solutions. The T1 and T2 results are shown in Table 3, and η is... Figure 1 .

[0060] Table 3 Effects of different concentrations of P407 on T1 and T2

[0061]

[0062] Experimental results show that, as shown in Table 3, when the contents of P188, PEG60001, 2- and propylene glycol remain constant, the gelation temperature is concentration-dependent on P407. With increasing P407 concentration, T1 decreases, T2 increases, and the gel range gradually widens. When the P407 concentration is 18%, the gelation temperature approaches the nasal cavity temperature.

[0063] Depend on Figure 1 It can be seen that the viscosity changes significantly with temperature, increasing noticeably near the gelation temperature T1. After gel formation, the viscosity remains relatively high with little change. When the P407 concentration is 16%, the gel range is short; after reaching the melting temperature T2, the gel transitions to a liquid state, and the viscosity decreases. As the P407 concentration increases, the gel range widens, and the viscosity gradually increases.

[0064] 2.2 Effects of P188 on T1, T2, and η

[0065] The prescribed amounts of paeonol-borneol nanoparticles, P407 18%, PEG6000 2%, and propylene glycol 5% were added to the drug solution. P188 was then added to prepare solutions of 0%, 1%, 2%, 3%, and 4% (w / w). T1 and T2 results are shown in Table 4, and η is... Figure 2 .

[0066] Table 4. Effects of different concentrations of P188 on T1 and T2

[0067]

[0068] Experimental results show that, as shown in Table 4, when the contents of P407, PEG6000, and 1,2-propanediol remain constant, both T1 and T2 increase with increasing P188 concentration. The gel range widens with increasing P188 concentration; however, there is no significant change in gel range at concentrations of 3% and 4%. At a P188 concentration of 2%, the gelation temperature approaches the nasal cavity temperature.

[0069] Depend on Figure 2 It can be seen that when the P188 concentration is 0% and 1%, the gel range is narrow. As the temperature increases, the formulation changes from liquid to gel and then back to liquid, and the viscosity changes significantly.

[0070] 2.3 Effects of PEG6000 on T1, T2, and η

[0071] The prescribed amounts of paeonol-borneol nanoparticles, P407 18%, P188 2%, and propylene glycol 5% were added to the drug solution. PEG was then added to prepare solutions of 0%, 1%, 2%, 3%, and 4% (w / w). The T1 and T2 results are shown in Table 5, and η is... Figure 3 .

[0072] Table 5. Effects of different concentrations of PEG6000 on T1 and T2.

[0073]

[0074]

[0075] Experimental results show that, as shown in Table 5, Figure 3 It can be seen that when the contents of P407, P188, and 1,2-propanediol remain constant, as the concentration of PEG6000 increases, T1 increases, T2 decreases, and the gel range gradually narrows. When the concentration of PEG6000 is 2%, the gelation temperature is close to the nasal cavity temperature.

[0076] Depend on Figure 3It can be seen that as the concentration of PEG6000 increases, the temperature required for a significant change in sample viscosity rises, and the gel range gradually narrows, meaning the temperature range for maintaining high viscosity narrows. When the concentration of PEG6000 increases to 4%, the viscosity decreases significantly. Increasing the concentration of PEG6000 reduces the adhesion of the formulation.

[0077] 2.4 Effects of propylene glycol on T1, T2, and η

[0078] The prescribed amounts of paeonol-borneol nanoparticles, P407 18%, P188 2%, and PEG6000 2% were added to the drug solution, and propylene glycol was added to prepare 2%, 3%, 4%, 5%, and 6% (w / w) solutions. The T1 and T2 results are shown in Table 6, and η is... Figure 4 .

[0079] Table 6. Effects of different concentrations on 1,2-propanediol T1 and T2.

[0080]

[0081] Experimental results show that, as shown in Table 6, when the contents of P407, P188, and PEG6000 remain constant, as the concentration of propylene glycol increases, T1 decreases, T2 increases, and the gel range gradually narrows. When the concentration of PEG6000 is 5%, the gelation temperature is close to the nasal cavity temperature.

[0082] Depend on Figure 4 It can be seen that the viscosity gradually increases with the increase of propylene glycol concentration. Propylene glycol has a certain viscosity, and its addition increases the viscosity.

[0083] In summary, the optimal formulation is as follows: P407 concentration of 18%, P188 concentration of 2%, PEG6000 concentration of 2%, and 1,2-propanediol concentration of 5%.

[0084] 3. Results of single-factor investigation

[0085] 3.1 Effect of the ratio of chitosan to paeonol feed mass on encapsulation efficiency

[0086] The results, as shown in Table 7, indicate that when the mass ratio of chitosan to paeonol was 6:1, the paeonol-borneol co-loaded nanoparticles exhibited the highest paeonol encapsulation efficiency. The paeonol encapsulation efficiency showed a trend of first increasing and then decreasing. This may be because when the amount of paeonol was higher, the amount of chitosan surrounding the paeonol decreased, leading to a reduction in the mechanical properties of the chitosan nanoparticles and a thinner coating around the drug, resulting in a poorer paeonol encapsulation effect, increased drug leakage, and ultimately, a decrease in encapsulation efficiency. However, as the chitosan content in the solution increased, the viscosity of the solution system increased, hindering the movement of paeonol around the chitosan molecules. Consequently, even with high chitosan content, the paeonol encapsulation efficiency also decreased.

[0087] Table 7. Mass ratio of chitosan and paeonol in feed

[0088]

[0089] 3.2 Effect of chitosan to sodium tripolyphosphate mass concentration ratio on encapsulation efficiency

[0090] The results are shown in Table 8. When the mass concentration ratio of chitosan to sodium tripolyphosphate is 3:1, the encapsulation efficiency of paeonol is relatively high. This may be because the positively charged amino groups of chitosan can form a stable ionic cross-linking network with the negatively charged sodium tripolyphosphate, which can effectively encapsulate the active drug while avoiding excessive cross-linking that leads to particle shrinkage. However, due to the high cross-linking density at this point, nanoparticles may aggregate due to electrostatic attraction or hydrophobic interactions, resulting in increased particle size, precipitation, and obvious stratification of the solution. When the mass concentration ratio of chitosan to sodium tripolyphosphate is greater than 4:1, the encapsulation efficiency of paeonol-borneol co-loaded nanoparticles shows a trend of first increasing and then decreasing with the increase of the chitosan to sodium tripolyphosphate mass concentration ratio. This may be because as the concentration of chitosan increases, its molecular surface carries more positive charge, thereby increasing the probability of paeonol being encapsulated. At the same time, the decrease in TPP content results in a sparser cross-linking network, smaller particles, more uniform dispersion, and improved solution transparency (reduced turbidity). However, if the concentration of chitosan solution is too high and TPP is extremely lacking, TPP will not be able to fully crosslink the amino groups of chitosan, resulting in fewer nanoparticles and difficulty in forming a stable network structure. The drug is very easy to leak, and thus the amount of paeonol that nanoparticles can encapsulate is reduced accordingly, and the encapsulation rate is lowered.

[0091] Table 8. Different mass concentration ratios of chitosan and sodium tripolyphosphate

[0092]

[0093] 3.3 Effect of chitosan solution pH on encapsulation efficiency

[0094] As shown in Table 9, when the pH is 3, the paeonol encapsulation efficiency of the paeonol-coated nanosuspension is relatively ideal, and it is not prone to stratification and is dispersed relatively uniformly. This may be because when the pH is less than 4, the H+ in the solution... + At higher concentrations, most of the NH2 in chitosan will be converted into NH3. + Furthermore, since they are all on the same molecular chain, and because they all carry the same type of charge, they generate a repulsive force that allows the molecular chain to extend, thus exposing more sites for interaction with the PO4+ of sodium tripolyphosphate. 3-Cross-linking occurs, encapsulating more paeonol and resulting in a higher encapsulation efficiency. However, at pH 2, due to the excessive positive charge of chitosan, it cannot polymerize well with the anions of TPP, thus reducing the encapsulation efficiency of paeonol. Furthermore, at pH 5 and pH 6, the paeonol-coated nano-suspension is relatively turbid, easily separates, and is unstable. This may be because at pH values ​​greater than 4, H… + As the concentration decreases, NH2 is converted into NH3. + The reduced number of -NH3 groups on the CS molecular chain leads to fewer cross-linking sites and fewer hydrogen bonds. Furthermore, a noticeable white flocculent precipitate appears in the solution when the pH value is greater than 4, indicating agglomeration and flocculation. These phenomena are primarily due to the reduced presence of -NH3 groups on the CS molecular chain. + This is caused by deprotonation. As pH increases, the -NH3 group on the CS molecular chain... + As the particles gradually transform into NH2, the charge on the nanoparticles gradually decreases, and the repulsive force between the particles gradually decreases. Under the action of stirring, the particles collide with each other, resulting in a significant increase in the size of the nanoparticles, which then form flocculent aggregates and precipitate.

[0095] Table 9 Chitosan solutions at different pH values

[0096]

[0097] 3.4 Effect of stirring speed on encapsulation efficiency

[0098] The results are shown in Table 10. The stirring speed had little effect on the encapsulation efficiency of paeonol, indicating that stirring speed generally does not affect the encapsulation efficiency. Furthermore, excessively high stirring speed can easily cause solution splashing, affecting experimental stability and subsequent volume measurements. Therefore, 1000 rpm was selected as the stirring speed for the magnetic stirrer, ensuring good uniform mixing of the solution without easily causing liquid splashing.

[0099] Table 10 Different stirring speeds

[0100]

[0101] 3.3 Results of Orthogonal Experiments

[0102] The results of the orthogonal experiment are shown in Table 11. A direct analysis of the range data shows that the order of influence of each factor on the preparation process is C > A > D > B. Meanwhile, the analysis of variance in Table 12 shows that factors A and C have a significant impact on the preparation process, while factor B has no significant impact. By comparing the k-values ​​of the three factors, the optimal formulation combination can be determined to be A1B2C2, namely, a CS-TPP mass concentration ratio of 6:1, a CS solution pH of 3, and a CS-(paeonol borneol) feed mass ratio of 4:1.

[0103] Table 11 Results of the orthogonal experiment

[0104]

[0105] Table 12 Analysis of Variance of Total Score

[0106] Note: * indicates a significant difference (P<0.05)

[0107] Compared with existing nasal drops on the market, the pharmaceutical composition of the present invention has significant clinical efficacy in treating allergic rhinitis, with high safety and improved quality of life for patients. Furthermore, the preparation method of the present invention is simple, easy to produce, and has broad market application prospects.

[0108] The following pharmacodynamic tests demonstrate the beneficial effects of this invention.

[0109] Experimental Example 1: Effect of the herbal composition of the present invention on IL-4-induced inflammatory factors in HNEpCs

[0110] This study demonstrates, through the IL-4-induced inflammatory cytokine profile, that HNEpCs are not only a physical barrier but also the "engine" of AR inflammation—driving the occurrence and persistence of allergic rhinitis through four major functions: Th2 polarization, barrier disruption, IgE upregulation, and Th1 inhibition.

[0111] 1. Experimental Materials

[0112] 1.1 Cells

[0113] Human nasal epithelial cells (HNEpCs) were purchased from BeiNa Biotechnology.

[0114] 1.2 Experimental reagents

[0115] The following formulas were prepared: a basic formula (Group A, 200g of Cynanchum paniculatum, 45g of Cicadae Periostracum, and 0.3g of Borneol), a patented formula (Group B, 200g of Cynanchum paniculatum, 45g of Cicadae Periostracum, 0.3g of Borneol, and 0.3g of Calculus Bovis), a traditional Chinese medicine composition of this invention (Group C, 200g of Cynanchum paniculatum, 45g of Cicadae Periostracum, 0.3g of Borneol, and 0.3g of Antrodia camphorata), and Antrodia camphorata (0.1, 0.2, 0.3, 0.4, and 0.5g) combined with the basic formula (Cynanchum paniculatum, Cicadae Periostracum, and Borneol). All the above formulas were extracted and concentrated with water to a concentration of 0.5mg / ml for later use.

[0116] 1.3 Establishment of an in vitro model of allergic rhinitis and grouping of patients for drug administration

[0117] An in vitro model of allergic rhinitis was established using IL-4-induced HNEpCs. Cells in the logarithmic growth phase were counted and diluted to the appropriate density for seeding in 6-well plates (1.4 × 10⁻⁶ cells per well). 5 1.8 × 10⁶ cells per well in a 24-well plate 40.4 × 10⁶ cells per well in a 96-well plate 4 When the growth density reaches about 80%, discard the culture medium and add blank or drug-containing culture medium to continue culturing.

[0118] Cells were divided into a control group, a model group (IL-4), and various drug groups. Based on the previous drug efficacy screening results, each dosage group was set at 0.5 mg / ml. The control group and the IL-4 group were added to blank culture medium containing 1% FBS, while each drug group was added with the corresponding drug. After culturing for 24 h, the model group and each drug group were added to IL-4 culture medium with a final concentration of 10 ng / ml. After culturing for 6 h, cells from each group were collected for later use.

[0119] 1.3 Detection of TNF-α, IL-1β, and IL-6 mRNA expression by qPCR

[0120] Total RNA was extracted from an appropriate amount of cells according to the kit instructions, and cDNA was obtained by reverse transcription using the RevertAid First strand cDNA Synthsis Kit. Then, PCR amplification was performed on the target genes (TNF-α, IL-1β, IL-6, GAPDH) to detect the mRNA expression levels of TNF-α, IL-1β, and IL-6. The amplification conditions were as follows: 50℃ preheating for 2 min, 95℃ preheating for 10 min, 95℃ denaturation for 15 s, 60℃ annealing for 30 s, and 60℃ extension for 30 s, for a total of 40 cycles. Results were analyzed using 2... －ΔΔCt Calculate the relative expression level of the target gene.

[0121] 1.4 Western blot analysis of iNOS and COX2 protein expression

[0122] After cell grouping and intervention, the culture medium was discarded, cells were washed with PBS, and the cells to be tested were collected. RIPA lysis buffer containing PMSF was added for lysis, and the lysis buffer was collected. The cells were centrifuged at 12000 rpm for 15 min at 4°C, and the supernatant was collected. Protein loading buffer was added to obtain protein samples. Proteins were separated on a prepared SDS-PAGE gel, followed by transfer and blocking. The cells were incubated overnight at 4°C with the target protein antibody, washed three times, and then incubated with the corresponding secondary antibody working solution at room temperature for 2 h. After three washes, images were developed using a Bio-RAD gel analyzer, and the corresponding protein expression was calculated using the grayscale values ​​of the protein bands.

[0123] 1.5 Statistical Analysis

[0124] Experimental data were analyzed using SPSS 20.0 software. All data in this study are presented as mean ± standard deviation. For data conforming to a normal distribution, one-way ANOVA was used among multiple groups. For homogeneous variances, the LSD test was used; for heterogeneous variances, the Games-Howell test was used. For data not conforming to a normal distribution, the nonparametric Mann-Whitney U test was used. P < 0.05 indicated statistical significance.

[0125] 2 Experimental Results

[0126] 2.1 Effects of drugs on IL-4-induced expression of TNF-α, IL-1β, and IL-6 mRNA in HNEpCs

[0127] like Figure 5 As shown, compared with the model IL-4 group, the expression of TNF-α, IL-1β, and IL-6 mRNA in each drug group was significantly reduced (P<0.01). Among them, compared with the basic formula (Xu Changqing, Cicadae Periostracum, Borneol) group A and group B, the expression of TNF-α and IL-1β mRNA in the traditional Chinese medicine composition (Xu Changqing, Cicadae Periostracum, Borneol, Antrodia camphorata) group C of this invention showed significant differences (P<0.05 or P<0.01).

[0128] like Figure 6 As shown, compared with the model IL-4 group, different combinations of Antrodia camphorata (0.1, 0.2, 0.3, 0.4, 0.5g) with basic formulas (Cynanchum paniculatum, Cicadae periostracum, Borneol) all reduced the expression of TNF-α, IL-1β, and IL-6 mRNA (P<0.05 or P<0.01). Among the different doses of Antrodia camphorata combination groups, 0.3g Antrodia camphorata showed the most significant reduction effect.

[0129] 2.2 Effects of drugs on IL-4-induced expression of iNOS and COX2 proteins in HNEpCs

[0130] like Figure 7 As shown, compared with the model IL-4 group, the expression of iNOS and COX2 proteins in each drug group was significantly reduced (P<0.01). Among them, compared with the basic formula (Xu Changqing, Cicadae Periostracum, Borneol) group A and group B, the expression of COX2 protein in the traditional Chinese medicine composition (Xu Changqing, Cicadae Periostracum, Borneol, Antrodia camphorata) group C of this invention was significantly different (P<0.05).

[0131] The above experiments demonstrate that the traditional Chinese medicine composition of the present invention has a more significant effect than the formulation disclosed in 201010238755.7.

Claims

1. A topical traditional Chinese medicine composition for treating allergic rhinitis, characterized in that: It is prepared from the following raw materials in the indicated weight ratios: Xu Changqing 160-240 parts, Cicadae Periostracum 36-54 parts, Borneol 0.24-0.36 parts, Antrodia camphorata 0.24-0.36 parts.

2. The topical traditional Chinese medicine composition for treating allergic rhinitis according to claim 1, characterized in that: It is prepared from the following raw materials in the indicated weight ratios: 200 parts of Cynanchum paniculatum, 45 parts of Cicadae periostracum, 0.3 parts of Borneol, and 0.3 parts of Antrodia camphorata.

3. The topical traditional Chinese medicine composition for treating allergic rhinitis according to claim 1 or 2, characterized in that: It is a commonly used topical pharmaceutical preparation made from paeonol extracted by Xu Changqing, water extracts of cicada molting and Antrodia camphorata, and borneol as active ingredients, with the addition of pharmaceutically acceptable excipients or auxiliary ingredients.

4. The topical traditional Chinese medicine composition for treating allergic rhinitis according to claim 3, characterized in that: The aforementioned topical preparations include nasal drops, nasal washes, nasal sprays, nasal aerosols, nasal ointments, nasal creams, and nasal gels.

5. The topical traditional Chinese medicine composition for treating allergic rhinitis according to claim 4, characterized in that: The nasal gel is prepared by extracting paeonol from Cynanchum paniculatum, decocting it with cicada molting and Antrodia camphorata to obtain a medicinal solution, then preparing borneol and paeonol into nanoparticles; adding propylene glycol, the medicinal solution, and a gel matrix.

6. The topical traditional Chinese medicine composition for treating allergic rhinitis according to claim 5, characterized in that: The nasal gel contains the following ingredients and matrix per 100 ml: 1.2g of nanoparticles, 5g of propylene glycol, 70ml of drug solution, plus 2g of poloxamer P188, 2g of polyethylene glycol PEG6000, 0.4g of potassium sorbate, and 18g of poloxamer P407; The 70ml of liquid contains 4.5g of raw cicada molting and 0.03g of Antrodia camphorata.

7. A method for preparing a topical traditional Chinese medicine composition for treating allergic rhinitis as described in claim 5 or 6, characterized in that: It includes the following steps: a. Preparation of the medicinal solution: Weigh the raw materials according to the weight ratio; extract the paeonol distillate by steam distillation of Cynanchum paniculatum, cool, settle, and filter to obtain crystals for later use; mix the extracted Cynanchum paniculatum residue with Cicadae Periostracum and Antrodia camphorata and decoct in water to extract, then mix the decoction with the remaining medicinal solution after steam distillation, filter and concentrate, centrifuge the concentrate, take the supernatant after concentration, add 1% chitosan solution, let stand and centrifuge, take the supernatant after centrifugation, concentrate the supernatant to obtain the medicinal solution; b. Preparation of nanoparticles: Borneol and paeonol were dissolved in PEG400 to form a borneol-paeonol solution. The solution was slowly added dropwise to a 2% chitosan acetate solution under magnetic stirring. The pH was adjusted to 3.0 with NaOH solution. Then, sodium tripolyphosphate (TPP) solution was slowly added dropwise under magnetic stirring. The solution was stirred continuously to obtain borneol-paeonol inclusion complex nanoparticles. c. Preparation of gel: Take the nanoparticles prepared in step b, add propylene glycol and drug solution and mix well, then add poloxamer P188, polyethylene glycol PEG6000, potassium sorbate and poloxamer P407, disperse them evenly, store at 4-10℃ overnight to obtain gel.

8. The method for preparing the topical traditional Chinese medicine composition for treating allergic rhinitis according to claim 7, characterized in that: In step b, the mass ratio of chitosan to paeonol is 6-8:1, preferably 6:1; the stirring speed is 1000 r; and the mass concentration ratio of chitosan to sodium tripolyphosphate is 6:

1.

9. Use of the traditional Chinese medicine composition according to any one of claims 1-6 in the preparation of a nasal medicament for treating allergic rhinitis.

10. The use according to claim 9, characterized in that: The aforementioned medication is used to treat allergic rhinitis caused by latent heat in the lung meridian.

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