A compound Chinese medicine composition for clearing throat and moistening sand thorn without water

Abstract

The application discloses a kind of water-free sand gorse compound throat-clearing and throat-moistening traditional Chinese medicine composition.The composition is calculated by raw drug and consists of the following components: 40-120 parts by weight of sea buckthorn leaves, 20-60 parts by weight of honeysuckle, 10-50 parts by weight of licorice, 1-3 parts by weight of sea buckthorn fruit powder, 0-40 parts by weight of dispersing agent, and 0-45 parts by weight of flavoring agent.The preparation method includes key steps such as ethanol solution extraction, reduced pressure concentration, freeze-drying, and wet granulation.The traditional Chinese medicine composition in the application produces a synergistic effect through scientific compounding.The in-vivo and in-vitro experiments show that the traditional Chinese medicine composition is significantly better than single-herb extract in inhibiting the production and release of NO in LPS-induced RAW264.7 cells;at the same time, it significantly reduces the number of inflammatory cell infiltrates in the peripheral blood of acute pharyngolaryngitis model rats, effectively repairs and improves pharyngeal mucosa damage, and shows excellent anti-inflammatory activity;and the prepared granules are quickly dissolved and melted when taken orally, and have excellent curative effect, taste, compliance, and process stability, and have industrialization prospects.

Description

Technical Field

[0001] This invention relates to the field of medicine, specifically a water-free, moisturizing sea buckthorn compound traditional Chinese medicine composition for clearing the throat and moisturizing the throat. Background Technology

[0002] Throat diseases are a global health problem with a continuously rising prevalence. Current market products mainly fall into two categories: traditional OTC drugs based on chemical agents or traditional Chinese medicine extracts, and functional foods such as compressed candies and herbal teas. However, both types of products have significant technical shortcomings and struggle to meet the needs of modern patients.

[0003] While traditional throat medications offer some therapeutic benefits, their application is significantly limited. Conventional formulations such as tablets, syrups, and capsules require water for administration, making them less suitable for elderly individuals with declining swallowing function or for mobile situations where medication administration is inconvenient. Furthermore, the inherent bitterness of traditional Chinese medicine formulas severely impacts patient palatability, leading to generally low adherence to traditional formulations among children and younger individuals. Those who use their voices extensively require convenient medication solutions that provide immediate relief without the need for drinking. Food-based products developed to improve taste often suffer from insufficient active ingredient content and ambiguous functional positioning, making it difficult to achieve definitive therapeutic effects and resulting in the long-standing technical dilemma of "effective if bitter, palatable if ineffective."

[0004] At the dosage form design and manufacturing process level, existing technologies also have shortcomings. On the one hand, traditional dosage forms fail to consider the physiological characteristics of special populations: swallowing dysfunction in the elderly can lead to medication risks, children have poor compliance with traditional dosage forms, and occupational users of the voice need medication solutions that provide immediate relief without relying on drinking water. On the other hand, existing traditional Chinese medicine preparation processes often have vague parameters, low extraction efficiency, unstable retention rates of active ingredients, and difficulty in achieving efficient transformation from laboratory to industrialization, resulting in large fluctuations in product quality and an inability to guarantee clinical efficacy.

[0005] In summary, existing technologies have consistently failed to overcome the systemic technical challenges across the four dimensions of "efficacy, taste, compliance, and process." The market urgently needs an innovative solution that simultaneously satisfies the requirements of definite efficacy, excellent taste, convenient administration, and stable processing—this is precisely the core technical problem that this invention aims to solve. Summary of the Invention

[0006] This invention aims to overcome multiple technical shortcomings of existing throat products, such as uncertain efficacy, poor taste, low patient compliance, and difficulty in industrializing production processes. Based on an in-depth analysis of the deficiencies of existing technologies, this invention provides a stable, safe, and effective traditional Chinese medicine formulation product through systematic formulation screening, process optimization, and dosage form innovation. This product combines definite antitussive and expectorant effects, heat-clearing and detoxifying properties, excellent palatability, extreme convenience with waterless sublingual administration, and a stable, controllable, and easily scalable preparation process, thereby comprehensively improving the user's medication experience and treatment continuity.

[0007] To achieve the above objectives, the present invention provides a water-free, moisturizing sea buckthorn compound traditional Chinese medicine composition for clearing the throat, characterized in that it comprises the following components based on the raw materials: 40-120 parts by weight of sea buckthorn leaves, 20-60 parts by weight of honeysuckle, 10-50 parts by weight of licorice, 1-3 parts by weight of sea buckthorn fruit powder, 0-40 parts by weight of dispersant, and 0-45 parts by weight of flavoring agent.

[0008] The components are in the following weight parts: 90 parts by weight of sea buckthorn leaves, 54 parts by weight of honeysuckle, 36 parts by weight of licorice, 1 part by weight of sea buckthorn fruit powder, 30 parts by weight of dispersant, and 41.25 parts by weight of flavoring agent.

[0009] The dispersant is selected from one or more of maltodextrin, starch, and mannitol; the flavoring agent includes a sweetener and a flavoring agent, wherein the sweetener is selected from one or more of erythritol and steviol glycosides, and the flavoring agent is selected from one or more of mint, jasmine, orange, peach, pear, and sweet orange.

[0010] The composition is a waterless sublingual granule that dissolves completely in 5 to 30 seconds after contact with saliva in the oral cavity.

[0011] The method for using the traditional Chinese medicine composition includes the following steps:

[0012] S1. Extraction: Weigh sea buckthorn leaves, honeysuckle, and licorice. Use 50%–100% ethanol aqueous solution as solvent and heat and reflux to extract 1–3 times at a material-to-liquid ratio of 1:(10–20), each time for 0.5–2 hours. Combine and filter to obtain a mixed extract.

[0013] S2. Concentration and drying: The mixed extract is concentrated under reduced pressure to 15% to 30% of its original volume, and then freeze-dried to obtain a dry extract;

[0014] S3. Molding: The dry paste is mixed with sea buckthorn fruit powder, dispersant and flavoring agent in equal increments, moistened with ethanol to form a soft material, wet granulated, and dried at low temperature to obtain granules.

[0015] In step S2, the reduced pressure concentration conditions are 40–45°C and -0.1–0.08 MPa; the freeze-drying conditions are vacuum freeze-drying, and the vacuum freeze-drying step involves pre-freezing the concentrate at -80°C for 12–24 h, reducing the vacuum degree to 60–100 Pa, gradually and slowly raising the drying temperature to 0°C, and continuing the drying time for 24–48 h.

[0016] In step S3, the moisture content of the soft material is controlled at 10% to 20%, the granulation screen is 16 to 24 mesh, and the drying temperature is 30 to 45°C.

[0017] The application of the aforementioned traditional Chinese medicine composition in the preparation of throat-soothing and moisturizing medicines.

[0018] The drug is used to treat acute and chronic pharyngitis, dry throat, itchy throat, cough, and phlegm.

[0019] The traditional Chinese medicine composition described above showed significantly better effects than single herb extracts in inhibiting LPS-induced NO production and release in RAW264.7 cells (P < 0.01, P < 0.05); it also significantly reduced the number of inflammatory cells in rats with acute pharyngitis and effectively repaired and improved pharyngeal mucosal damage. Its anti-inflammatory effect was superior to that of commercially available control drugs Huasu tablets, diisocyanate lozenges, and Ryukakusan granules (P < 0.01).

[0020] The dosage forms of the traditional Chinese medicine composition also include clinically required dosage forms such as tablets, pills, capsules, and oral liquids.

[0021] The traditional Chinese medicine composition of this invention is based on the scientific compatibility of traditional Chinese medicine theory, wherein the formulation principle and the efficacy of each medicinal material are as follows:

[0022] Guided by traditional Chinese medicine theory, this invention scientifically combines sea buckthorn leaves (principal ingredient), honeysuckle (assistant ingredient), licorice root (guide ingredient), and sea buckthorn fruit powder. Traditional throat-clearing formulas often use heat-clearing and detoxifying herbs as the principal ingredient; this invention uses warm-natured sea buckthorn leaves as the principal ingredient, combined with cooling honeysuckle, reflecting the TCM principle of "using both warming and clearing herbs." In vitro experiments have demonstrated that the compound of this invention significantly inhibits LPS-induced NO release more effectively than any single herb (P<0.01), and the high-dose group significantly reduces inflammatory markers such as white blood cells and lymphocytes in a rat model of acute pharyngitis, showing significantly better results than commercially available Huasu tablets and Ryukakusan (P<0.01), producing a synergistic effect that was unforeseen by those skilled in the art.

[0023] This formula uses sea buckthorn leaves, which are warm in nature, promote blood circulation, remove blood stasis, and have antibacterial and anti-inflammatory properties, as the principal ingredient, targeting the root cause of throat diseases. Honeysuckle, which clears heat, drains fire, dries dampness, and detoxifies, is used as the assistant ingredient to enhance the heat-clearing and detoxifying effects of the principal ingredient. Licorice is added as the guiding ingredient, which not only harmonizes the other ingredients but also exerts its unique heat-clearing, cough-relieving, and phlegm-reducing effects. Sea buckthorn fruit powder improves the taste while supplementing nutrition and strengthening the body. The combined effects of these ingredients are to strengthen the body's resistance, relieve cough and phlegm, and invigorate the spleen and resolve dampness. This can enhance the body's immunity and resist viral invasion, and is suitable for symptoms of throat discomfort such as dry throat, itchy throat, and pharyngitis.

[0024] Hippophae rhamnoides L., a plant belonging to the genus Hippophae in the family Elaeagnaceae, is widely distributed in Northwest my country and possesses significant ecological and economic value. Sea buckthorn leaves are a novel food resource rich in bioactive substances, containing flavonoids, tannins, polysaccharides, vitamins, and other active ingredients, exhibiting pharmacological activities such as anti-inflammatory, antioxidant, anti-aging, immune-boosting, and antibacterial effects. Sea buckthorn berries are a traditional Chinese medicine resource with both medicinal and edible uses; they can be used as an auxiliary ingredient to improve taste and also possess rich nutritional value. Sea buckthorn berries are rich in vitamin C, vitamin E, carotene, unsaturated fatty acids, and other nutrients, and also contain flavonoids similar to those in sea buckthorn leaves, which have synergistic immune-boosting, anti-inflammatory, and antioxidant effects.

[0025] The preferred content of honeysuckle in traditional Chinese medicine combination products is 20-60 parts by weight, and more preferably 54 parts by weight. Honeysuckle is derived from the dried flower buds or newly opened flowers of Lonicera japonica Thunb., a plant of the Caprifoliaceae family. It mainly contains organic acids, flavonoids, volatile oils, and iridoids, possessing pharmacological effects such as antipyretic, antioxidant, anti-inflammatory, and immune-enhancing properties. Honeysuckle can both dispel wind-heat and clear blood toxins, making it effective for various febrile diseases, such as fever, rashes, spots, carbuncles, and sore throat. Acute inflammation is another important manifestation of "heat toxin" syndrome, and honeysuckle, with its significant anti-inflammatory effects, plays an important role in the prevention and treatment of diseases in traditional Chinese medicine.

[0026] Licorice is the dried root and rhizome of the legumes *Glycyrrhiza uralensis* Fisch., *Glycyrrhiza inflata* Bat., or *Glycyrrhiza glabra* L., and is used as the adjuvant in the formula of this invention. This medicinal material is sweet and neutral in nature, and enters the spleen, stomach, lung, and heart meridians. It has the effects of tonifying the spleen and replenishing qi, resolving phlegm and relieving cough, alleviating spasms and pain, and harmonizing other herbs. It is effective in treating various types of cough with phlegm, whether due to cold, heat, deficiency, or excess. Its main active ingredients include flavonoids (such as glycyrrhizin) and triterpenoids (such as glycyrrhizic acid). In this formula, licorice not only enhances the cough-relieving and phlegm-resolving effects but also harmonizes the medicinal properties of sea buckthorn leaves and honeysuckle, reducing the potential impact of the cold nature of honeysuckle on the spleen and stomach.

[0027] Modern pharmacological studies have shown that sea buckthorn leaves, sea buckthorn berries, honeysuckle, and licorice, as natural plant components, are rich in various active ingredients such as flavonoids, saponins, polysaccharides, and alkaloids, all of which have demonstrated significant potential in the treatment of acute pharyngitis. The abundant bioactive components in these four herbs provide a solid material foundation for their development into functional foods for clearing and moisturizing the throat. This invention, through extensive experimental research, has confirmed that the combination of sea buckthorn leaves, honeysuckle, and licorice produces a significant synergistic effect. This synergistic effect is not only reflected in the rationality of the combination according to traditional Chinese medicine theory but has also been verified through modern pharmacological experiments.

[0028] The present invention discloses a waterless, throat-soothing compound traditional Chinese medicine composition of sea buckthorn, which has the following beneficial effects: by combining sea buckthorn leaves (principal drug), honeysuckle (assistant drug), and licorice (guide drug), a synergistic effect is produced, which is significantly better than the extracts of each single herb (P < 0.01, P < 0.05). Confirmed in a rat model of acute pharyngitis, the high-dose group is significantly better than the commercially available control drug in inhibiting inflammatory cells (such as leukocytes, monocytes, lymphocytes, and neutrophils) (P < 0.01). The waterless granules prepared by compound flavoring technology have excellent taste and dissolve rapidly in saliva upon contact in the oral cavity, eliminating the need for water administration and greatly improving the convenience and compliance of medication. At the same time, the preparation process parameters of the present invention are clear, stable, and controllable. By optimizing key steps such as extraction, freeze-drying, and granulation using an equal-volume incremental method, the active ingredients are effectively preserved, ensuring uniform and stable product quality and possessing excellent industrial scale-up potential. This invention successfully solves the technical dilemma of simultaneously achieving efficacy, taste, compliance, and process stability. Attached Figure Description

[0029] Figure 1 This is a schematic diagram of the preparation method of the traditional Chinese medicine combination product provided by the present invention;

[0030] Figure 2 The chromatogram of the test solution of the traditional Chinese medicine combination product;

[0031] Figure 3 The appearance and dissolution state of the three groups of particles are shown;

[0032] Figure 4 RAW264.7 cell viability and NO inhibition rate after drug administration;

[0033] Figure 5 The changes in the pharynx of rats in each group after modeling and drug administration;

[0034] Figure 6 The changes in the main blood routine indicators of rats in each group after modeling and drug administration are shown. Detailed Implementation

[0035] Example 1

[0036] Weigh out 120 parts by weight of cleaned and impurity-removed sea buckthorn leaves, 60 parts by weight of honeysuckle, and 50 parts by weight of licorice root, and mix them to obtain a total of 230 parts by weight of medicinal materials. Place the medicinal materials in an extraction tank, add 12 times the volume (i.e., 2760 parts by weight) of a 70% ethanol aqueous solution, and heat under reflux for extraction twice, 1.5 hours each time. Combine the extracts, filter, and obtain a mixed extract.

[0037] The mixed extract was concentrated under reduced pressure at 45°C and -0.08 MPa to about 25% of its original volume, and then freeze-dried to obtain the traditional Chinese medicine extract powder. The extraction performance of each traditional Chinese medicine composition preparation example is shown in Table 1.

[0038] Take 5 parts by weight of the above extract powder, 2 parts by weight of sea buckthorn fruit powder, 20 parts by weight of starch, 20 parts by weight of erythritol, 0.1 parts by weight of peppermint flavoring and 4 parts by weight of peach flavoring, mix them together using the equal incremental method, add 70% ethanol to make a soft material, granulate it through a 16-mesh sieve, dry it at 40℃ and then granulate it to obtain granules.

[0039] Example 2

[0040] Weigh out 100 parts by weight of sea buckthorn leaves, 50 parts by weight of honeysuckle, and 40 parts by weight of licorice, for a total of 190 parts by weight of medicinal materials. Add 10 times the amount (1900 parts by weight) of 75% ethanol aqueous solution, and extract by heating and reflux three times, each time for 1.5 hours. Subsequent concentration and drying steps are the same as in Example 1.

[0041] Take 5 parts by weight of the above-mentioned extract powder, 1.5 parts by weight of sea buckthorn fruit powder, 10 parts by weight of starch, 15 parts by weight of erythritol, 0.2 parts by weight of peppermint flavoring and 5 parts by weight of orange flavoring, mix them together, and granulate them according to the method of Example 1 to obtain granules.

[0042] Example 3

[0043] Weigh out 80 parts by weight of sea buckthorn leaves, 40 parts by weight of honeysuckle, and 30 parts by weight of licorice, for a total of 150 parts by weight of medicinal materials. Add 14 times the amount (2100 parts by weight) of 75% ethanol aqueous solution, and extract twice by heating and reflux for 0.5 hours each time. Subsequent concentration and drying steps are the same as in Example 1.

[0044] Take 5 parts by weight of the above-mentioned extract powder, 3 parts by weight of sea buckthorn fruit powder, 25 parts by weight of starch, 35 parts by weight of erythritol, 0.15 parts by weight of peppermint flavoring and 4.5 parts by weight of orange flavoring, mix them together, and granulate them according to the method of Example 1 to obtain granules.

[0045] Example 4

[0046] Weigh out 60 parts by weight of sea buckthorn leaves, 30 parts by weight of honeysuckle, and 20 parts by weight of licorice, for a total of 110 parts by weight of medicinal materials. Add 10 times the amount (1100 parts by weight) of 80% ethanol aqueous solution, and extract twice by heating and reflux, 1 hour each time. Subsequent concentration and drying steps are the same as in Example 1.

[0047] Take 5 parts by weight of the above-mentioned extract powder, 1 part by weight of sea buckthorn fruit powder, 35 parts by weight of starch, 25 parts by weight of erythritol, 0.3 parts by weight of peppermint flavoring and 3.8 parts by weight of pear flavoring, mix them together, and granulate them according to the method of Example 1 to obtain granules.

[0048] Example 5

[0049] Weigh out 90 parts by weight of sea buckthorn leaves, 54 parts by weight of honeysuckle, and 36 parts by weight of licorice, for a total of 180 parts by weight of medicinal materials. Add 10 times the amount (1800 parts by weight) of 70% ethanol aqueous solution, and extract by heating and reflux three times, each time for 1.5 hours. Subsequent concentration and drying steps are the same as in Example 1.

[0050] Take 5 parts by weight of the above-mentioned extract powder, 1 part by weight of sea buckthorn fruit powder, 30 parts by weight of starch, 30 parts by weight of erythritol, 0.35 parts by weight of peppermint flavoring, 5 parts by weight of orange flavoring and 4.9 parts by weight of jasmine flavoring, mix them together, and granulate them according to the method of Example 1 to obtain granules.

[0051] Table 1 Comparison of leaching performance of various traditional Chinese medicine compositions in preparation examples

[0052]

[0053] Note: M1 = M quercetin + isorhamnetin + kaempferol, M2 = M glycyrrhizin + glycyrrhizic acid, M3 = M chlorogenic acid + isochlorogenic acid A + isochlorogenic acid C + luteolin, M4 = M 总黄酮 M5=M 总浸膏

[0054] See Figure 1 , Figure 1 This is a schematic diagram of the preparation method of the traditional Chinese medicine combination product provided by the present invention.

[0055] Example 6

[0056] Qualitative analysis of Example 5 of the above-mentioned traditional Chinese medicine combination product is as follows:

[0057] HPLC chromatographic conditions: Column: GS-120-5-C18-AP (4.6 mm ID × 250 mm); Mobile phase: Acetonitrile (A) - 0.4% phosphoric acid aqueous solution (B); Gradient elution program as follows: 0-6 min, 8-11% A; 6-7 min, 11-12% A; 7-9 min, 12-13% A; 9-19 min, 13% A; 19-30 min, 13-14% A; 30-40 min, 14-20% A; 40-44 min, 20-26% A; 44-45 min, 26-30% A; 45-59 min, 30-37% A; 59-63 min, 37-41% A; 63-65 min, 41-55% A; 65-66 min, 55-100% A. A; 66-76 min, 100% A; 76-77 min, 100-8% A; 77-87 min, 8% A; Detection wavelength: 260 nm; Flow rate: 0.7 mL / min; Column temperature: 30℃; Injection volume: 10 μL.

[0058] Preparation of reference solution: Take an appropriate amount of reference standard, accurately weigh it, and dissolve it in methanol to obtain the solution.

[0059] Preparation of the test solution: Weigh 2 g of this product accurately, dissolve it in 10 mL of distilled water, extract it three times with 20 mL of water-saturated n-butanol solution, concentrate to remove the n-butanol solution, dissolve it in 5 mL of methanol, and the test solution is obtained.

[0060] Accurately pipette 10 μL each of the reference solution and the test solution into the liquid chromatograph and determine their properties.

[0061] Record the chromatogram.

[0062] See Figure 2 , Figure 2 This is a chromatogram of the test solution of a traditional Chinese medicine combination product.

[0063] Example 7

[0064] Accurately weigh 50 mg of the traditional Chinese medicine composition, add 1200 μL of 70% methanol for ultrasonic extraction, centrifuge (12000 rpm, 15 min), aspirate the supernatant, filter the sample through a 0.22 μm microporous membrane, and store it in an injection vial for UPLC-QTOF-MS / MS analysis. Separation was performed on an ACQUITY UHPLC C18 column (2.1 × 50 mm, 1.7 μm, Waters, USA), with mobile phase A being 0.1% formic acid aqueous solution and mobile phase B being 0.1% formic acid acetonitrile. Gradient elution was performed as follows: 0-3 min, 5% B; 3-4 min, 5-9% B; 4-15 min, 9-14% B; 15-21 min, 14-60% B; 21-22 min, 60-100% B; 22-23 min, 100% B; 23-24 min, 100-5% B; 24-28 min, 5% B. Injection volume: 2 µL; flow rate: 0.3 mL / min; column temperature: 36℃. Mass spectrometry analysis conditions were as follows: electrospray ionization (ESI), negative ion detection mode, scan range: m / z 100–1700. The ESI source parameters are as follows: dry gas (N2 flow rate of 5 mL / min, dry gas temperature of 300℃), atomizing gas pressure of 35 psi, capillary voltage of 3500 V, nozzle voltage of 1000 V, capillary outlet voltage of 400 V, and collision energy of 20 eV.

[0065] By comparing with standards, databases, and references, a total of 88 compounds were identified in the components of traditional Chinese medicine, as shown in Table 2.

[0066] Example 8

[0067] (1) 20 parts by weight of the Chinese herbal extract powder obtained by freeze-drying and pulverizing in Example 5 were directly added to ethanol to wet and make soft material. The soft material was then granulated by wet method and dried at low temperature. The granules were then subjected to sensory evaluation and solubility test.

[0068] (2) Take 5 parts by weight of the Chinese herbal extract powder obtained by freeze-drying and pulverizing in Example 5 and 30 parts by weight of the dispersant, add ethanol to wet and make soft material, granulate by wet method, dry at low temperature, and perform sensory evaluation and solubility test on the granules.

[0069] (3) Sensory evaluation and solubility test were performed on the particles obtained in Example 5.

[0070] Figure 3 The sensory evaluation and solubility test results for the three groups of particles are shown below:

[0071] Granules made without any excipients are dark brown in color, have a sticky and greasy texture, poor solubility, and a strong bitter taste from the herbs themselves, resulting in poor compliance and affecting the user experience.

[0072] The granules made after adding the dispersant are light brown in color and the solubility is greatly improved, but the bitter taste is still obvious, which affects the overall taste.

[0073] The granules obtained from the Chinese medicine combination product in Example 5 are light yellow in color. After dissolving, they are clear and bright, with a cool and pleasant taste. They dissolve rapidly after being taken orally. The granules begin to dissolve almost immediately upon contact with saliva and dissolve completely in a short time without the need for additional chewing or waiting.

[0074] Experimental Example 1: In vitro experiment using RAW264.7 cells

[0075] 1. Experimental Methods

[0076] (1) Preparation of test solution

[0077] Accurately weigh 20 parts by weight each of sea buckthorn leaves, honeysuckle, and licorice, and prepare individual herbal granules using the process described in Example 5: sea buckthorn granules (HG), honeysuckle granules (LG), and licorice granules (GG), as well as the herbal composition granules from Example 5 (CSOG). Take appropriate amounts of HG, LG, GG, and CSOG respectively, and dissolve them thoroughly in DMSO to obtain a 200 mg / mL test solution.

[0078] (2) MTT cell viability assay

[0079] RAW264.7 cells were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C, 5% CO2, and 95% relative humidity. RAW246.7 cells in the logarithmic growth phase were harvested and cultured at a rate of 9 × 10⁻⁶ cells / year. 4 Cells were seeded at a density of 200 μL per well in 96-well plates and cultured for 4 hours until cell attachment. Drug treatment was then administered. The experiment included a control group and a sample group. The control group received no drug, while the sample groups received single-herb granules (HG, LG, GG) at a final concentration of 500 μg / mL and CSOG granules at final concentrations of 250, 500, and 1000 μg / mL, respectively, with four replicates per group. After drug treatment, cells were cultured for another 24 hours. 100 μL of supernatant from each well was discarded, and 10 μL of MTT solution was added. After incubation for 4 hours, 100 μL of triplet solution was added to each well, and the cells were allowed to stand overnight to fully dissolve the formazan crystals. The absorbance of each well was measured at 570 nm using a microplate reader, and the viability of RAW246.7 cells was calculated.

[0080] (3) NO inhibition rate determination

[0081] RAW246.7 cells in the logarithmic growth phase were harvested and cultured at a concentration of 9 × 10⁻⁶ cells / year. 4 Cells were seeded at a density of 200 μL per well in 96-well plates and cultured for 4 hours until cell adhesion was achieved before drug administration. The experiment included a blank control group, a sample group, a positive control group, and a model group. The blank control group received no drug. The sample group received single-herb granules (HG, LG, GG) at a final concentration of 500 μg / mL, and CSOG granules at final concentrations of 250, 500, and 1000 μg / mL, respectively. The model group received LPS at a final concentration of 1 μg / mL, and the positive control group received quercetin at a final concentration of 10 μg / mL. Each group had four replicates. After drug administration, cells were cultured for another 0.5 hours, then LPS at a final concentration of 1 μg / mL was added, and incubation continued for 24 hours. 100 μL of the supernatant was collected, and 100 μL of Griess reagent was added. The absorbance of each well was measured at 540 nm using a microplate reader, and the NO inhibition rate was calculated.

[0082] 2. Experimental Results

[0083] Figure 4 To assess the viability of RAW264.7 cells and the NO inhibition rate after drug administration, the in vitro anti-inflammatory activity and cellular safety of each test sample were evaluated by detecting the viability of RAW264.7 cells and the NO inhibition rate after different drug administration treatments.

[0084] After treatment with various drugs, the survival rate of RAW264.7 cells ranged from 93.39% to 109.90%, indicating that none of the samples had significant cytotoxic effects within this concentration range. When evaluating the inhibitory effect on LPS-induced NO release, the results showed that the NO level in the cell supernatant of the model group was significantly increased compared with the normal control group (P < 0.01), suggesting that LPS successfully induced the establishment of an inflammation model. All treatment groups inhibited the production and release of LPS-mediated inflammatory mediators to varying degrees. Among them, at a concentration of 500 μg / mL, the inhibitory effect of the traditional Chinese medicine composition granules on NO was significantly better than that of the granules prepared from the extracts of the three single herbs (P < 0.05; P < 0.01), and this inhibitory effect showed a clear dose-dependent relationship (compared with quercetin: **P, ****P < 0.01; compared with CSOG-500: ## P, ### P, #### P < 0.01.

[0085] Table 2. Identification of compounds in traditional Chinese medicine compositions

[0086]

[0087]

[0088] Note: Q: Quercetin, K: Kaempferol, I: Isorhamnetin, S: Syringetin, L: Laricitrin, HHDP: hexahydroxydiphenoyl., * indicates reference substance identification; the rest are identified by LC-MS.

[0089] Experimental Example 1 In Vivo Experiment on Rats with Acute Pharyngolaryngitis

[0090] 1. Experimental Materials and Methods

[0091] Forty-two SPF-grade SD rats, male, weighing 140 ± 20 g, were provided by the Animal Experiment Center of Lanzhou University, with the license number SYXK (Gan) 2023-0004. After being adaptively fed for 1 week under standard laboratory conditions, the rats were randomly divided into 7 groups, with 6 rats in each group:

[0092] Table 3 Grouping Table of Experimental Doses for Rats in Each Group

[0093] Except for the blank group, the rats in the other groups were established with acute pharyngolaryngitis models by ammonia water spraying method: 0.2 mL of ammonia water was sprayed on the throat, twice a day for 3 consecutive days. After successful model establishment, each drug administration group was given corresponding drug intervention for 7 consecutive days. The administration dose was converted to an equivalent dose according to the body surface area.

[0094] 2. Observation Indexes

[0095] (1) General behavioral observation: Observe the mental state, activity level, fur color and luster, respiratory state, whether there is behavior of scratching the neck, changes in food and water intake, body weight changes, coughing and wheezing conditions, etc. of the rats every day;

[0096] (2) Blood routine detection: After the end of model establishment and after the last administration, collect the blood samples of the rats, and use Mindray veterinary automatic hematology analyzer (BC-2800vet) to detect the changes in the number of inflammatory cells (white blood cells, monocytes, lymphocytes, and neutrophils);

[0097] 3. Experimental Results

[0098] 3.1 General Behavioral Observation

[0099] As Figure 5 shown, before model establishment, during the adaptive feeding period of the rats in each group, the overall mental state was good, the diet and water intake were normal, the hair color was white and shiny, the pharyngeal mucosa was intact and light red, the mouth and nose were clean without abnormal secretions, and the body weight increased normally.

[0100] During the modeling period, compared with the control group, the modeled rats showed significantly reduced activity, poor mental state, dull fur, frequent drinking, reduced food intake, and weight loss. As the number of days of modeling increased, the modeled rats gradually exhibited frequent scratching of their mouth and nose, redness and swelling of the skin around the mouth and nose with scratch marks, loss of fur around the mouth, increased mucus secretion in the throat, congestion and swelling of the pharyngeal mucosa with a bright red color, and coughing symptoms such as "face-washing" movements, head nodding, hoarseness, abdominal muscle twitching, and rapid breathing.

[0101] During the administration period, rats in the traditional Chinese medicine composition group showed a significant reduction in scratching of their mouth and nose, as well as a significant decrease in oral and nasal secretions. Their fur color and activity levels also improved significantly, lost fur around the mouth began to recover, and their body weight increased. While the commercially available tablets showed improvement, the effect was not as significant as that of the high-dose traditional Chinese medicine composition group. In the model group, rats showed no significant change in body weight, activity level, or food and water intake, and still exhibited symptoms of coughing and scratching of their mouth and nose.

[0102] 3.2 Blood routine test results

[0103] The number of white blood cells, lymphocytes, monocytes, and neutrophils in the model group rats was significantly higher than that in the control group (P<0.01), indicating that the model was successfully established. Figure 6 As shown (compared with the model group after modeling: *P < 0.05, **P, ***P, ****P < 0.01; compared with the blank group after drug administration: #### P < 0.01.

[0104] The inflammatory indicators of the Chinese herbal medicine combination group were significantly lower than those of the model group (P<0.01), and significantly lower than those of the commercially available Ryukakusan group and Huasu tablet group (P<0.01), indicating that there is indeed a synergistic effect.

[0105] Figure 6 The data analysis shown in the chart is as follows:

[0106] (1) The charts represent a complete inflammatory response assessment system, demonstrating the anti-inflammatory effects of the product of this invention from different dimensions:

[0107] White blood cell count: reflects the overall level of inflammation in the body;

[0108] Lymphocytes: reflect the strength of a specific immune response;

[0109] Monocytes: Reflect chronic inflammation and tissue repair status;

[0110] Neutrophils: Reflect the degree of acute inflammatory response.

[0111] (2) Detailed analysis of sub-indicators

[0112] a. White blood cell count analysis

[0113] After modeling: The white blood cell count in the model group (Mod) increased significantly (approximately 25 × 10⁻⁶). 9 The presence of / L indicates that the acute pharyngitis model has been successfully established, and the body exhibits a severe systemic inflammatory response.

[0114] After administration: The high-dose group (H) of this invention showed the most significant effect, restoring the white blood cell count to near-normal levels (approximately 10 × 10⁻⁶). 9 / L); dose-response relationship is clear: low (L), medium (M), high (H) doses

[0115] The dosage group showed a gradient improvement; it was superior to the positive control: the effect was significantly better than Huasu tablets (HSP) and Ryukakusan (LJS);

[0116] b. Lymphocyte analysis

[0117] Clinical significance: Overactivation of lymphocytes is an important mechanism of chronic inflammation and immune damage.

[0118] Effects of this invention: The high-dose group can significantly reduce the number of lymphocytes, indicating that it can not only control acute inflammation, but also regulate abnormal immune responses, which is of great significance for preventing the chronicity of pharyngitis.

[0119] c. Mononuclear cell analysis

[0120] Clinical significance: Monocytes are precursors to tissue macrophages and participate in chronic inflammation and tissue repair.

[0121] Effects of this invention: The high-dose group significantly reduced the number of monocytes, indicating that the product can inhibit the process of chronic inflammation and prevent acute inflammation from developing into a chronic stage.

[0122] d. Neutrophil analysis

[0123] Clinical significance: Neutrophils are the primary responders in acute inflammation, and their number directly reflects the severity of acute inflammation.

[0124] The high-dose group showed the most significant inhibitory effect on neutrophils, indicating that the product of this invention can quickly control acute inflammation and rapidly relieve acute symptoms such as redness, swelling, and pain.

[0125] Experimental Example 2: Evaluation of Solubility and Taste

[0126] The granules prepared in Examples 1-5 were tested according to the test methods for granules in the General Rules for Preparations of the Chinese Pharmacopoeia 2020. Ten trained evaluators scored the granules from five aspects: texture, odor, color, taste, and solubility, with a full score of 100. The sensory quality scoring criteria are shown in Table 4.

[0127] Table 4 Sensory Quality Scoring Criteria for Particles

[0128] The results are shown in Table 5:

[0129] Table 5. Sensory Evaluation Comparison Table of Preparation Examples of Various Traditional Chinese Medicine Compositions

[0130]

Claims

1. A waterless, moisturizing sea buckthorn compound traditional Chinese medicine composition for clearing the throat and moisturizing the throat, characterized in that, Based on the raw material, it is composed of the following components: 40-120 parts by weight of sea buckthorn leaves, 20-60 parts by weight of honeysuckle, 10-50 parts by weight of licorice, 1-3 parts by weight of sea buckthorn fruit powder, 0-40 parts by weight of dispersant, and 0-45 parts by weight of flavoring agent.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The components are in the following weight parts: 90 parts by weight of sea buckthorn leaves, 54 parts by weight of honeysuckle, 36 parts by weight of licorice, 1 part by weight of sea buckthorn fruit powder, 30 parts by weight of dispersant, and 41.25 parts by weight of flavoring agent.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The dispersant is selected from one or more of maltodextrin, starch, and mannitol; the flavoring agent includes a sweetener and a flavoring agent, wherein the sweetener is selected from one or more of erythritol and steviol glycosides, and the flavoring agent is selected from one or more of mint, jasmine, orange, peach, pear, and sweet orange.

4. The traditional Chinese medicine composition according to claim 1, characterized in that, The composition is a waterless sublingual granule that dissolves completely in 5 to 30 seconds after contact with saliva in the oral cavity.

5. A method for preparing the traditional Chinese medicine composition according to any one of claims 1-4, characterized in that, Includes the following steps: S1. Extraction: Weigh sea buckthorn leaves, honeysuckle, and licorice. Use 50%–100% ethanol aqueous solution as solvent and heat and reflux to extract 1–3 times at a material-to-liquid ratio of 1:(10–20), each time for 0.5–2 hours. Combine and filter to obtain a mixed extract. S2. Concentration and drying: The mixed extract is concentrated under reduced pressure to 15% to 30% of its original volume, and then freeze-dried to obtain a dry extract; S3. Molding: The dry paste is mixed with sea buckthorn fruit powder, dispersant and flavoring agent in equal increments, moistened with ethanol to form a soft material, wet granulated, and dried at low temperature to obtain granules.

6. The method according to claim 5, characterized in that, In step S2, the reduced pressure concentration conditions are 40–45°C and -0.1–0.08 MPa; the freeze-drying conditions are vacuum freeze-drying, and the vacuum freeze-drying step involves pre-freezing the concentrate at -80°C for 12–24 h, reducing the vacuum degree to 60–100 Pa, gradually and slowly raising the drying temperature to 0°C, and continuing the drying time for 24–48 h.

7. The method according to claim 5, characterized in that, In step S3, the moisture content of the soft material is controlled at 10% to 20%, the granulation screen is 16 to 24 mesh, and the drying temperature is 30 to 45°C.

8. The use of the traditional Chinese medicine composition according to any one of claims 1-4 in the preparation of a throat-soothing and moisturizing medicine.

9. The application as described in claim 8, characterized in that, The drug is used to treat acute and chronic pharyngitis, dry throat, itchy throat, cough, and phlegm.

10. The traditional Chinese medicine composition according to any one of claims 1-4, characterized in that, This traditional Chinese medicine composition inhibits LPS-induced NO production and release in RAW264.7 cells.

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