Traditional Chinese medicine granule for treating flat wart and preparation method thereof

Abstract

The invention discloses traditional Chinese medicine granules for treating verruca plana and a preparation method thereof. The traditional Chinese medicine granules comprise traditional Chinese medicine dry extract powder and auxiliary materials, the auxiliary materials comprise a diluent, a wetting agent and stevioside; wherein the traditional Chinese medicine dry extract powder is extracted from the following components in parts by weight: 60 parts of radix isatidis, 60 parts of semen coicis, 60 parts of herba portulacae, 30 parts of radix arnebiae seu lithospermi, 20 parts of semen persicae, 20 parts of flos carthami, 60 parts of herba equiseti hiemalis, 20 parts of rhizoma cyperi, 60 parts of nacre mother of pearl, 20 parts of honeycomb and 60 parts of raw oyster shell. The prepared granules are uniform in particle size and good in quality, the water content ranges from 1.59% to 1.79%, the forming rate ranges from 96.0% to 96.9%, the angle of repose ranges from 26.82 degrees to 29.99 degrees, the comprehensive performance score is high, and repeatability is good.

Description

Technical Field

[0001] This invention relates to a traditional Chinese medicine composition and its preparation method, and more particularly to a traditional Chinese medicine granule for treating flat warts and its preparation method, belonging to the field of traditional Chinese medicine technology. Background Technology

[0002] Flat warts are a common viral skin disease caused by human papillomavirus (HPV) infection. In Traditional Chinese Medicine (TCM), they fall under the category of "flat warts" and are often attributed to internal heat and toxins, and stagnation of qi and blood. This disease is characterized by recurrent episodes and a prolonged, difficult-to-cure course, making clinical treatment challenging. TCM treatment for flat warts often employs internal, external, or combined methods, but existing formulas frequently suffer from complex formulations, inconvenient application, and a high recurrence rate after initial treatment.

[0003] In the research of traditional Chinese medicine (TCM) preparations for flat warts, oral liquids and topical washes are available dosage forms. However, the preparation of oral liquids typically involves complex processes such as decoction, collection of distillate, and concentration, making the preparation steps cumbersome and the finished product bulky, inconvenient to carry and store. Topical washes are mainly applicable to local skin lesions and have limited effect on regulating systemic symptoms or deeper pathogenesis. In addition, liquid dosage forms generally suffer from relatively poor stability, susceptibility to mold growth, and unpleasant taste, which may affect patients' long-term medication adherence. Compared with traditional prescriptions or conventional Western topical medications, existing dosage forms still have room for improvement in terms of convenience, stability, and patient adaptability.

[0004] Therefore, developing a modern dosage form that is easy to prepare, has controllable quality, high stability, and is convenient for patients to take, such as granules, is of great significance for fully leveraging the therapeutic advantages of this compound and improving its clinical applicability. Summary of the Invention

[0005] Purpose of the invention: The purpose of this invention is to provide a traditional Chinese medicine granule for treating flat warts that has high efficacy, good quality, and good reproducibility; another purpose of this invention is to provide a method for preparing the traditional Chinese medicine granule.

[0006] Technical Solution: This invention provides a traditional Chinese medicine granule for treating flat warts. The granule comprises a dry extract powder of traditional Chinese medicine and excipients. The excipients include a diluent, a wetting agent, and steviol glycosides. The dry extract powder of traditional Chinese medicine is extracted from the following formula in parts by weight: 60 parts of Isatis root, 60 parts of Coix seed, 60 parts of Portulaca oleracea, 30 parts of Lithospermum erythrorhizon, 20 parts of peach kernel, 20 parts of safflower, 60 parts of Equisetum hyemale, 20 parts of Cyperus rotundus, 60 parts of mother-of-pearl, 20 parts of bee honeycomb, and 60 parts of raw oyster shell.

[0007] The aforementioned technical solution involves a traditional Chinese medicine composition for treating flat warts. Its fixed prescription consists of 60 parts Isatis root, 60 parts Coix seed, 60 parts Portulaca oleracea, 30 parts Lithospermum erythrorhizon, 20 parts Peach kernel, 20 parts Safflower, 60 parts Equisetum hyemale, 20 parts Cyperus rotundus, 60 parts Pearl powder, 20 parts Bee honeycomb, and 60 parts Raw oyster shell. This compound uses Isatis root as the principal ingredient to clear heat and detoxify; Coix seed, Portulaca oleracea, and Lithospermum erythrorhizon are the assistant ingredients to assist in clearing heat and promoting blood circulation; Peach kernel, Safflower, Equisetum hyemale, and Cyperus rotundus are the adjuvant ingredients to promote blood circulation, remove blood stasis, soothe the liver, and relieve depression; Pearl powder, Bee honeycomb, and Raw oyster shell are the guiding ingredients to soften and disperse nodules, and attack toxins and fight viruses. The high dosage combination of Pearl powder and Bee honeycomb, combined with Raw oyster shell, forms a dual-enhancing effect of "softening and dispersing nodules + attacking toxins and fighting viruses," which is the core innovation of this compound. The compound has been verified to show good application potential in the treatment of flat warts.

[0008] Preferably, the mass ratio of dry extract powder to excipients is 1:2-2.5, and the amount of steviol glycosides added is 0.4-0.6% of the total mass of excipients.

[0009] Preferably, the diluent consists of lactose and mannitol; the wetting agent is ethanol with a concentration of 85-95%.

[0010] Preferably, the diluent consists of lactose and mannitol in a mass ratio of 2-1:1.

[0011] The method for preparing the traditional Chinese medicine granules for treating flat warts according to the present invention includes the following steps:

[0012] (1) Soak raw oysters and calcined mother-of-pearl in water, boil them and then filter to obtain the filtrate. The filtrate obtained is the decoction extract.

[0013] (2) The decoction extract, raw oyster residue, calcined mother-of-pearl residue and other 9 medicinal materials are combined, soaked in water, decocted and filtered, centrifuged, and the supernatant is collected. The supernatant is concentrated under reduced pressure to extract, dried under reduced pressure and then pulverized and sieved to obtain dry extract powder.

[0014] (3) The dry extract powder and excipients are mixed evenly, and the Chinese medicine granules are prepared by wet granulation process.

[0015] Preferably, in step (1), raw oysters and calcined mother-of-pearl are soaked in 8-10 times the total amount of water for 1-2 hours, decocted for 0.5-1.5 hours, and then filtered to obtain the filtrate. The decoction and filtration are repeated 1-3 times, and the filtrates are combined to obtain the first decoction extract.

[0016] More preferably, in step (1), raw oysters and calcined mother-of-pearl are soaked in 9-10 times the total amount of water for 1-1.5 hours, decocted for 0.5-1.5 hours, and then filtered to obtain filtrate. The decoction and filtration are repeated 2-3 times, and the filtrates are combined to obtain the first decoction extract.

[0017] Most preferably, in step (1), raw oysters and calcined mother-of-pearl are soaked in 10 times the amount of water of the total medicinal materials for 1.5 h, decocted for 0.5 h, and then filtered to obtain filtrate. The decoction and filtration are repeated 3 times, and the filtrates are combined to obtain the first decoction extract.

[0018] Preferably, in step (2), the decoction extract, raw oyster residue, calcined mother-of-pearl residue and other 9 medicinal materials are combined, and soaked in 8-10 times the total amount of water of the other 9 medicinal materials for 1.5-2.5 hours. After decoction for 0.75-2 hours, the filtrate is obtained. The decoction and filtration are repeated 1-3 times. After centrifugation, the supernatant is collected. The supernatant is concentrated under reduced pressure to extract. After drying under reduced pressure, it is pulverized and sieved to obtain dry extract powder.

[0019] More preferably, in step (2), the decoction extract, raw oyster residue, calcined mother-of-pearl residue and other 9 medicinal materials are combined, and soaked in 9-10 times the total amount of water of the other 9 medicinal materials for 2-2.5 hours. After decoction for 1-2 hours, the filtrate is obtained. The decoction and filtration are repeated 2-3 times. After centrifugation, the supernatant is concentrated under reduced pressure to extract. After drying under reduced pressure, it is pulverized and sieved to obtain dry extract powder.

[0020] Most preferably, in step (2), the decoction extract, raw oyster residue, calcined mother-of-pearl residue and other 9 medicinal materials are combined, and soaked in 9 times the total amount of water of the other 9 medicinal materials for 1.5 hours. After decoction for 1 hour, the filtrate is obtained. The decoction and filtration are repeated twice. After centrifugation, the supernatant is concentrated under reduced pressure to extract. After drying under reduced pressure, it is pulverized and sieved to obtain dry extract powder.

[0021] Preferably, in step (3), the dry powder and excipients are mixed evenly, granulated, dried, and sized to obtain Chinese medicine granules.

[0022] More preferably, in step (3), the drying temperature is 50-70°C.

[0023] Most preferably, in step (3), the drying temperature is 60°C.

[0024] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: The granules prepared by the present invention have uniform particle size and good quality, with a water content between 1.59-1.79, a molding rate between 9.60-9.69, an angle of repose between 26.82-29.99, a high comprehensive performance score, and good repeatability. Attached Figure Description

[0025] Figure 1 Thin-layer chromatograms for the identification of Isatis indigotica root. From left to right in the figure, they are test sample 1, test sample 2, test sample 3, (R,S)-Gouyichun reference standard, Isatis indigotica root reference material, and negative control without Isatis indigotica root.

[0026] Figure 2Thin-layer chromatograms for identification of Coix seed, from left to right: test sample 1, test sample 2, test sample 3, Coix seed oil reference extract, Coix seed reference medicinal material, and negative control without Coix seed;

[0027] Figure 3 The thin-layer chromatograms for the identification of purslane are shown from left to right as follows: test sample 1, test sample 2, test sample 3, purslane reference material, and negative control without purslane. Detailed Implementation

[0028] The technical solution of the present invention will be further described below.

[0029] Example 1

[0030] This traditional Chinese medicine compound formula consists of: Isatis root 60g, Coix seed 60g, Portulaca oleracea 60g, Lithospermum erythrorhizon 30g, Peach kernel 20g, Safflower 20g, Equisetum hyemale 60g, Cyperus rotundus 20g, Mother-of-pearl 60g, Bee honeycomb 20g, and Raw oyster shell 60g. Preparation method: First decoction: Decoction of mother-of-pearl and raw oyster shell, adding water to cover the herbs, decocting for 0.5 hours; Second decoction: Add the remaining herbs, adding water to cover all herbs, decoct three times (first 2 hours, second 1 hour, third 1 hour), collect approximately 250ml of distillate after boiling; combine the filtrates, let stand, collect the supernatant, concentrate to an appropriate volume; add the distillate, add water to adjust to 400ml, mix well, filter, and obtain the oral liquid. Dosage: Oral administration, twice a day, 50ml each time. Use for 6 weeks. Verification showed that the traditional Chinese medicine compound in this embodiment achieved a 100% efficacy rate and a 0% recurrence rate for treating flat warts. Adjustments to the dosage of the herbs in this embodiment did not achieve the same therapeutic effect. The mother-of-pearl, honeycomb, and raw oyster shell in this embodiment have a synergistic effect; all three are indispensable, and the absence of any one of these herbs in the compound will significantly reduce the efficacy.

[0031] Example 2

[0032] The extraction process for this embodiment was screened as follows:

[0033] 1. Determination of dry extract yield and content

[0034] Determination of dry extract yield: Refer to the "Extractives Determination Method" in the 2020 edition of the Chinese Pharmacopoeia: Take 20 mL of extract into an evaporating dish (dried to constant weight), evaporate to dryness in a water bath, dry in an oven at 105℃ for 3 h to constant weight, then cool in a desiccator for 30 min, accurately weigh the mass, and calculate the dry extract yield.

[0035] Dry extract yield = [(extract mass × total sample volume) / (material mass × sample quantity)] × 100%

[0036] The quality of the dried extract refers to the quality of the extract after drying; the quality of the processed medicinal materials refers to the quality of the medicinal materials before extraction begins.

[0037] Content determination:

[0038] Content = C × V × N / M

[0039] Where C is the concentration of the index component in the extract, mg / mL; V is the total volume of the extract, mL; N is the dilution factor; and M is the mass of the medicinal powder, g.

[0040] In the extraction process of this compound, high performance liquid chromatography was used to detect the contents of uridine, guanosine and adenosine.

[0041] 2. Single-factor investigation of pre-cooking

[0042] 2.1 Examination of soaking time

[0043] Weigh 1 / 5 of the prescribed amount of calcined mother-of-pearl and raw oyster shell, and weigh five portions in parallel. Soak for 1 hour, 1.5 hours, 2 hours, 2.5 hours, and 3 hours, adding 6 times the amount of water. Decoction time: 30 minutes. Extract once. Filter the extract and record the total volume. Dry in an evaporating dish to constant weight. Take 30 ml of the extract and concentrate in a water bath. After concentration to dryness, place in an oven at 105°C and continue drying to constant weight. (Cool in a desiccator for 30 minutes.) Quickly and accurately weigh the extract. Calculate its dry extract yield.

[0044] Table 1. Effect of different soaking times on the yield of pre-decocted dry extract.

[0045]

[0046] The table shows that soaking time affects the pre-decoction extraction rate of calcined mother-of-pearl and raw oyster shells. The highest dry extract yield was obtained when the soaking time was 1 hour. As the soaking time increased, the dry extract yield decreased but tended to stabilize. Therefore, orthogonal design experiments were conducted with soaking times of 1 hour, 1.5 hours, and 2 hours to determine the optimal pre-decoction extraction process.

[0047] 2.2 Investigation of the water addition ratio

[0048] Weigh 1 / 5 of the prescribed amount of calcined mother-of-pearl and raw oyster shell, and weigh five portions in parallel. Using the optimal soaking time determined in the previous experiment, add 6, 7, 8, 9, and 10 times the amount of water respectively, and decoct for 30 minutes, extracting once. Filter the extract and record the total volume. Dry in an evaporating dish to constant weight. Take 30 ml of the extract and concentrate in a water bath in an evaporating dish. After concentrating to dryness, place in an oven at 105°C and continue drying to constant weight. (Cool in a desiccator for 30 minutes.) Quickly and accurately weigh the extract. Calculate its dry extract yield.

[0049] Table 2. Effect of different water addition ratios on the yield of pre-decocted dry extract.

[0050]

[0051] The water addition ratio affects the pre-decoction extraction rate of calcined mother-of-pearl and raw oyster shell. As the water addition ratio increases, the dry extract yield initially increases and then decreases. The highest dry extract yield of calcined mother-of-pearl and raw oyster shell is achieved at a water addition ratio of 9. Therefore, orthogonal design experiments were conducted with water addition ratios of 8, 9, and 10 to determine the optimal pre-decoction extraction process.

[0052] 2.3 Examination of extraction time

[0053] Weigh 1 / 5 of the prescribed amount of calcined mother-of-pearl and raw oyster shell, and take five parallel portions. Using the optimal soaking time and optimal water addition ratio determined in the previous experiment, extract for 0.5h, 1h, 1.5h, 2h, and 2.5h respectively, once. Filter the extract and record the total volume. Dry in an evaporating dish to constant weight. Take 30ml of the extract and concentrate in a water bath in an evaporating dish until dry. Then, place it in an oven at 105℃ and continue drying to constant weight (cool in a desiccator for 30 minutes). Quickly and accurately weigh the extract. Calculate its dry extract yield.

[0054] Table 3. Effects of different decoction times on the yield of pre-decocted dry extract.

[0055]

[0056] The table shows that extraction time affects the pre-decoction extraction rate of calcined mother-of-pearl and raw oyster shell. With increasing decoction time, the dry extract yield initially increases and then slowly decreases. The highest dry extract yield is achieved at an extraction time of 1 hour. Therefore, orthogonal design experiments were conducted with extraction times of 0.5 hours, 1 hour, and 1.5 hours to determine the optimal pre-decoction extraction process.

[0057] 2.4 Examination of the number of extractions

[0058] Weigh 1 / 5 of the prescribed amount of calcined mother-of-pearl and raw oyster shell, and take five parallel portions. Extract 1, 2, and 3 times respectively using the optimal soaking time, optimal decoction time, and optimal water addition ratio selected in the previous experiment. Filter the extract and record the total volume. Dry the extract in an evaporating dish to constant weight. Take 30 ml of the extract and concentrate it in a water bath in an evaporating dish. After concentrating to dryness, place it in an oven at 105°C and continue drying to constant weight. (Cool in a desiccator for 30 minutes.) Quickly and accurately weigh the extract. Calculate its dry extract yield.

[0059] Table 4. Effect of different extraction times on the yield of the pre-decoction extract.

[0060]

[0061] The table shows that the number of extractions affects the pre-extraction rate of calcined mother-of-pearl and raw oyster shells; the dry extract yield increases with the number of extractions. Considering cost savings in industrial production, only the third extraction was selected. Therefore, an orthogonal design experiment was conducted with extractions of 1, 2, and 3 times to determine the optimal pre-extraction process.

[0062] 3. Orthogonal experiment with decoction first

[0063] Based on the results of the single-factor investigation above, an orthogonal experiment was conducted on the factors that significantly affected the experimental results.

[0064] Table 5. Factor Level Table for Extraction Process

[0065]

[0066] Table 6 Orthogonal Experiment

[0067]

[0068] Table 7. Analysis of Variance Table

[0069]

[0070] Intuitive analysis: The orthogonal experimental design shows that R(D) > R(B) > R(C) > R(A), meaning the number of extractions > the water addition ratio > extraction time > soaking time. The average values ​​of the four factors at different levels are, in descending order: A2 > A3 > A1, B3 > B2 > B1, C1 > C3 > C2, D3 > D2 > D1. The range data indicates that the optimal extraction process is A2B3C1D3, which involves soaking for 1.5 hours, adding 10 times the water volume, boiling for 0.5 hours, and extracting 3 times.

[0071] Analysis of variance (ANOVA): The ANOVA table shows that the order of influence of the four factors on the extraction process is consistent with the intuitive analysis, namely, extraction times > water addition ratio > extraction time > soaking time. Using the dry extract yield as the evaluation index, the ANOVA table shows that factors B and D have a certain degree of significance (P<0.05), meaning that the water addition ratio and extraction times have a significant impact on the dry extract yield, while other factors have a relatively small impact.

[0072] Comprehensive analysis: Based on the results of orthogonal experiment verification, intuitive analysis, and variance analysis, the optimal extraction process was determined to be A2B3C1D3, which means soaking for 1.5 hours, adding 10 times the amount of water, extracting for 0.5 hours, and extracting 3 times.

[0073] 4. Verification of the pre-frying process

[0074] To verify the accuracy and reproducibility of the optimal process conditions selected by orthogonal experiments, a confirmatory experiment was conducted. Three parallel portions (1 / 5 of the prescription amount) of calcined mother-of-pearl and raw oyster shell were weighed, and the yield of the dry extract was determined using the previously selected optimal soaking time, optimal extraction time, optimal water addition ratio, and optimal number of extractions.

[0075] Table 8 Verification Table of Pre-frying Process

[0076]

[0077] Three parallel experiments were conducted to verify the extraction conditions described above. The extraction was performed three times: calcined mother-of-pearl and raw oyster were soaked for 1.5 hours, then 10 times the amount of water was added, and the extraction was carried out for 0.5 hours. The RSD of the dry extract was measured to be 0.35%, indicating that the orthogonal design was reasonable.

[0078] 5. Single-factor analysis of co-decoction extraction

[0079] 5.1 Examination of soaking time

[0080] Take 1 / 5 of the prescribed amount of Isatis root, stir-fried peach kernel, coix seed, horsetail, bee honeycomb, purslane, safflower, gromwell root, and vinegar-processed Cyperus rhizome. Weigh five portions in parallel, and soak them for 1 hour, 1.5 hours, 2 hours, 2.5 hours, and 3 hours respectively. Combine the extracts of calcined mother-of-pearl and raw oyster shell (previously decocted) and the dregs with the other herbs, add water to 8 times the volume, and decoct for 2 hours. Decoction once. Collect the volatile oil. Determine the contents of uridine, guanosine, and adenosine using high-performance liquid chromatography (HPLC), and determine the yield of the dry extract.

[0081] Overall weighted average score = (Dry extract yield / Highest dry extract yield) × 25 + (Uridine content / Highest uridine content) × 25 + (Guanine content / Highest guanine content) × 25 + (Adenosine content / Highest adenosine content) × 25

[0082] Table 9. Effects of different soaking times on co-decoction extraction

[0083]

[0084] The table shows that soaking time affects the extraction rate of the compound Isatis root extract. The highest comprehensive score of the extract was obtained when the soaking time was 2 hours. As the soaking time increased, the content and the dry extract rate decreased. Therefore, orthogonal design experiments were conducted with soaking times of 1.5 hours, 2 hours, and 2.5 hours to determine the optimal co-decoction extraction process.

[0085] 5.2 Investigation of the water addition ratio

[0086] Take 1 / 5 of the prescribed amount of Isatis root, stir-fried peach kernel, coix seed, horsetail, bee honeycomb, purslane, safflower, gromwell root, and vinegar-processed Cyperus rhizome. Weigh five portions in parallel. Using the optimal soaking time determined in the previous experiment, combine the extracts of calcined mother-of-pearl and raw oyster shell, along with the dregs, with the other medicinal materials. Add water to 6, 7, 8, 9, and 10 times the volume, respectively. Decoction time: 2 hours, once. Collect the volatile oil. Determine the content of uridine, guanosine, and adenosine, and the yield of the dry extract.

[0087] Table 10 Effect of different water addition ratios on co-decoction extraction

[0088]

[0089] As shown in the table, the extracts of the compound medicinal materials exhibited the highest contents of uridine, guanosine, and adenosine, as well as the highest overall score of the dry extract, when the water addition ratio was 9 times. Therefore, further orthogonal design experiments were conducted with water addition ratios of 8, 9, and 10 times to determine the optimal co-decoction extraction process.

[0090] 5.3 Examination of extraction time

[0091] Take 1 / 5 of the prescribed amount of Isatis root, stir-fried peach kernel, coix seed, horsetail, bee honeycomb, purslane, safflower, gromwell root, and vinegar-processed Cyperus rhizome. Weigh five portions in parallel. Using the optimal soaking time determined in the previous experiment, combine the extracts of calcined mother-of-pearl and raw oyster shell, along with the dregs, with the other medicinal materials. Add the optimal water volume determined in the previous experiment. Extract for 1 hour, 1.5 hours, 2 hours, 2.5 hours, and 3 hours, respectively, and decoct once. Collect the volatile oil. Determine the content of uridine, guanosine, and adenosine, and the yield of the dry extract.

[0092] Table 11 Effect of different extraction times on co-decoction extraction

[0093]

[0094] The table shows that the highest overall extraction rate score was achieved when the extraction time was 1 hour. Further orthogonal design experiments were conducted with extraction times of 0.75 hours, 1 hour, and 1.25 hours to determine the optimal co-decoction extraction process.

[0095] 5.4 Examination of the number of extractions

[0096] Take 1 / 5 of the prescribed amount of Isatis root, stir-fried peach kernel, coix seed, horsetail, bee honeycomb, purslane, safflower, gromwell root, and vinegar-processed Cyperus rhizome. Weigh five portions in parallel. Using the optimal soaking time determined in the previous experiment, combine the pre-decocted extracts of calcined mother-of-pearl and raw oyster shell with the dregs and other medicinal materials. Add the optimal amount of water determined in the previous experiment and extract 1, 2, and 3 times using the optimal extraction time determined in the previous experiment. Collect the volatile oil. Determine the content of uridine, guanosine, and adenosine, and the yield of the dry extract.

[0097] Table 12 Effect of different extraction times on co-decoction extraction

[0098]

[0099] As shown in the table, the combined scores for uridine, guanosine, adenosine, and dry extract yield were highest when extracted twice. Therefore, an orthogonal design experiment was conducted to select extraction times of 1, 2, and 3 times to determine the optimal co-decoction extraction process.

[0100] 6. Orthogonal experiment of co-decoction

[0101] Press L9(3) 4 An orthogonal array was used to conduct experiments, and the uridine, guanosine, and adenosine content and the yield of the dry extract in the nine aqueous extracts were calculated. The results were then analyzed using a comprehensive weighted average method.

[0102] Overall weighted average score = (Dry extract yield / Highest dry extract yield) × 25 + (Uridine content / Highest uridine content) × 25 + (Guanine content / Highest guanine content) × 25 + (Adenosine content / Highest adenosine content) × 25

[0103] Table 13 Factor Level Table for Extraction Process

[0104]

[0105] Table 14 Orthogonal Experiment

[0106]

[0107] Table 15 Analysis of Variance Table

[0108]

[0109] Intuitive analysis: The orthogonal experimental design shows that R(D) > R(C) > R(A) > R(B), meaning the number of extractions > extraction time > soaking time > water addition ratio. The average values ​​of the four factors at different levels are, in descending order: A1 > A3 > A2, B2 > B3 > B1, C2 > C1 > C3, D2 > D1 > D3. The range data indicates that the optimal extraction process is A1B2C2D2, which involves soaking for 1.5 hours, adding 9 times the amount of water, extracting for 1 hour, and extracting twice.

[0110] Analysis of variance (ANOVA): The ANOVA table shows that the order of influence of the four factors on the extraction process is consistent with the intuitive analysis, namely, extraction times > extraction time > soaking time > water addition ratio. Using the contents of uridine, guanosine, and adenosine, and the dry extract yield as the evaluation indicators, the ANOVA table shows that factors A, C, and D have a certain degree of significance (P<0.05), meaning that soaking time, extraction time, and extraction times have a significant impact on the contents of uridine, guanosine, and adenosine, as well as the dry extract yield, while the water addition ratio has a relatively small impact.

[0111] Comprehensive analysis: Based on the results of orthogonal experiment verification, intuitive analysis, and variance analysis, the optimal extraction process was determined to be A1B2C2D2, which is soaking for 1.5 hours, adding 9 times the amount of water, extracting for 1 hour, and extracting twice.

[0112] 7. Validation of co-decoction extraction process

[0113] To verify the accuracy and reproducibility of the optimal process conditions selected by orthogonal experiments, a confirmatory experiment was conducted. Five portions of the following herbs were taken in parallel: Isatis root, roasted peach kernel, coix seed, horsetail, bee honeycomb, purslane, safflower, gromwell root, and vinegar-processed cyperus rhizome. Using the previously selected optimal soaking time and water ratio, the extracts and residues from the pre-decocted calcined mother-of-pearl and raw oyster shell were combined with the soaked herbs. The previously selected optimal decoction time and number of extractions were then used. The volatile oil was collected. The total content of uridine, guanosine, and adenosine, as well as the dry extract yield, were determined and calculated using the above method.

[0114] Table 16 Validation of the co-decoction extraction process

[0115]

[0116] As shown in the table above, the RSDs of the total content and yield of the three batches of samples were 3.02% and 2.84%, respectively, with good reproducibility. Therefore, A1B2C2D2 was determined, which means soaking for 1.5 hours, adding 9 times the amount of water, extracting for 1 hour, and extracting twice.

[0117] Example 3

[0118] Commonly used granulation processes include dry granulation and wet granulation. Since dry granulation has high requirements for both the drug and excipients, and wet granulation has advantages such as uniform particle size and good quality, wet granulation is used in this example.

[0119] The formulation screening for the granules in this embodiment is as follows:

[0120] 1. Evaluation Methods

[0121] 1.1 Determination of molding rate

[0122] Take particles that can pass through sieve No. 1 but not sieve No. 5, weigh them and record the weight as the mass of the particles after sieving, and calculate the molding rate.

[0123] Formation rate (%) = (Mass of particles after sieving / Mass of particles before sieving) × 100%

[0124] 1.2 Determination of hygroscopicity

[0125] Take a supersaturated sodium chloride solution and place it in a glass desiccator. Then, place the desiccator in a 25°C constant temperature incubator for 24 hours. Spread the granules evenly on the bottom of a weighing bottle that has been dried to constant weight. After weighing, open the cap of the weighing bottle, place it in the glass desiccator, and put it in a 25°C constant temperature incubator. After 6 hours, remove it and weigh it.

[0126] Moisture absorption rate (%) = (Mass of particles after moisture absorption - Weight of particles before moisture absorption) / (Mass of particles before moisture absorption) × 100%

[0127] 1.3 Scoring Criteria

[0128] Table 17 Scoring Criteria

[0129]

[0130] 2. Prescription Screening Research

[0131] 2.1 Investigation of wetting agent concentration

[0132] Five portions of the 1 / 5 prescription amount of medicinal materials were weighed out, and volatile oil and distillate were extracted by steam distillation. The volatile oil was collected, and the distillate was concentrated and dried. The resulting dry powder was mixed with lactose at a ratio of 1:2 (steviosides accounted for 0.2% of the total excipients). Ethanol solutions with concentrations of 75%, 80%, 85%, 90%, and 95% were used to prepare a soft mass until it reached a state where it "could be formed into a ball when squeezed, but crumbled when pinched." The soft mass was extruded through a 14-mesh sieve to form wet granules. The wet granules were dried at 60℃ (moisture content 3%~5%), and the dried granules were sieved through a 14-mesh sieve for granulation and mixed to obtain granules composed of three different excipients. The effect of different wetting agent concentrations on granulation was investigated.

[0133] Table 18 Investigation of Ethanol Concentration

[0134]

[0135] The overall score for ethanol concentration = (soft material condition × 50%) + (sieve quality × 50%)

[0136] Based on the above investigation of ethanol concentration, 85% ethanol was ultimately selected as the wetting agent for further investigation.

[0137] 2.2 Investigation of diluents

[0138] Dextrin, lactose, and mannitol are commonly used excipients in granule preparations. Therefore, dextrin, lactose, and mannitol were compared. Following the methods in the Chinese Pharmacopoeia (2020 edition), the softness, solubility, forming rate, angle of repose, and moisture absorption of the granules were observed. The comprehensive score was used as a reference standard to screen out the optimal excipients and their optimal formulation.

[0139] Weigh out 3 portions of dry powder separately, mix the obtained dry powder with dextrin, mannitol and lactose in a 1:2 ratio (steviosides account for 0.2% of the total excipients), select the ethanol solution selected above, prepare a soft material until the soft material reaches the state of "forming a ball when squeezed and crumbling when pinched", extrude the soft material through a 14-mesh sieve to make wet granules, dry the wet granules at 60℃ in time, granulate the dried granules through a 14-mesh sieve, mix well, and the product is obtained.

[0140] Table 19. Investigation of diluents

[0141]

[0142] The overall score for fillers is calculated as follows: Solubility × 20% + Softness × 20% + Formability × 20% + Flowability × 20% + Hygroscopicity × 20%.

[0143] Based on the above considerations regarding particle solubility, soft material properties, angle of repose, molding rate, and hygroscopicity, mannitol and lactose were ultimately selected as the optimal diluents for further investigation into mixed diluents.

[0144] 2.3 Investigation of Mixed Diluents

[0145] Five portions of dry powder were weighed separately. The optimal ratio of the two previously screened optimal excipients was investigated using a mixing diluent at ratios of 3:1, 2:1, 1:1, 1:2, and 1:3. The dry powder and excipients were mixed at a ratio of 1:2 (steviosides accounted for 0.2% of the total excipients). An ethanol solution selected earlier was used to prepare a soft mass until it reached a state where it "could be formed into a ball when squeezed, but crumbled when pinched." The soft mass was then extruded through a 14-mesh sieve to form wet granules. The wet granules were dried at 60°C immediately. The dried granules were then sieved through a 14-mesh sieve to form larger granules and mixed thoroughly. The soft mass, solubility, formability, angle of repose, and moisture absorption of the granules were observed, and their comprehensive scores were used as a reference standard to screen the best excipients and optimal ratios.

[0146] Table 20. Investigation of mixed fillers

[0147]

[0148] The overall score for the mixed diluent = solubility × 20% + softness × 20% + moldability × 20% + flowability × 20% + hygroscopicity × 20%

[0149] Based on the above investigations into particle solubility, soft material properties, angle of repose, molding rate, and hygroscopicity, lactose:mannitol 2:1 was ultimately selected as the optimal mixing diluent for further investigation into the drug-excipient ratio.

[0150] 2.4 Investigation of the drug-excipient ratio

[0151] Five portions of dry extract powder were weighed and mixed with the previously screened optimal mixed excipients and dry extract powder in ratios of 1:1, 1:1.5, 1:2, 1:2.5, and 1.5:1 (steviosides accounted for 0.2% of the total excipients). An ethanol solution was selected from the previously screened mixture to prepare a soft mass until it reached a state where it "could be formed into a ball when grasped, but crumbled when squeezed." The soft mass was then extruded through a 14-mesh sieve to form wet granules. The wet granules were dried immediately at 60°C. The dried granules were then sieved through a 14-mesh sieve to form uniform granules and mixed thoroughly. The solubility, soft mass condition, sieving results, angle of repose, and formability were scored to select the appropriate amount of excipients.

[0152] Table 21 Investigation of Auxiliary Material Usage

[0153]

[0154] The overall score for auxiliary material usage is calculated as follows: Solubility × 20% + Softness × 20% + Sieving Quality × 20% + Flowability × 20% + Formability × 20%.

[0155] Based on the above examinations of particle solubility, softness, sieving, angle of repose, and forming rate, the ratio of dry powder to excipients of 1:2 yielded the highest overall score and the best particle size. Therefore, a drug-excipient ratio of 1:2 was selected as the optimal ratio for further investigation.

[0156] 2.5 Investigation into the Dosage of Flavoring Agents

[0157] The taste of oral preparations can affect their drug quality and patient medication adherence. Adding flavoring agents to traditional Chinese medicine preparations can improve the off-flavor and bitterness of traditional Chinese medicine granules, making them more acceptable to patients.

[0158] Since the taste of the granules obtained from this formula is poor before the addition of sweetener, a certain amount of flavoring agent needs to be added to improve the taste of the medicine. Considering the clinical application for patients with diabetes, obesity and other conditions, and because steviol glycosides are stable to heat, acid and alkali and have good safety, steviol glycosides were chosen as the flavoring agent for this formula.

[0159] Weigh out 3 portions of dry powder and mix them with the previously selected optimal mixed excipients and dry powder according to the previously selected optimal drug-excipient ratio. Add steviol glycosides, accounting for 0.2%, 0.4%, 0.6%, and 0.8% of the total excipients, respectively. Select the previously selected ethanol solution to make a soft material until the soft material reaches the state of "forming a clump when squeezed but crumbling when pinched". The soft material is squeezed through a 14-mesh sieve to make wet granules. Dry the wet granules at 60°C in time. After drying, granulate through a 14-mesh sieve to form granules and mix well to obtain the final product.

[0160] Ten subjects were selected, and they were scored according to the above scoring criteria. The final average score was calculated.

[0161] Table 22 Taste scores based on the amount of steviol glycosides used

[0162]

[0163] As shown in the table, 0.6% steviol glycosides are more popular among consumers. Therefore, 0.6% steviol glycosides were chosen for further research.

[0164] 2.6 Investigation of drying temperature

[0165] To prevent excessive drying time from causing the granules to easily absorb moisture and deliquesce, the effects of drying time on granule formation rate, angle of repose, solubility, and moisture content were investigated.

[0166] Weigh out 3 portions of dry powder, select the optimal mixed excipients selected above, mix the dry powder and excipients according to the proportions selected above, select the ethanol solution selected above, and make a soft material until the soft material reaches the state of "forming a ball when squeezed and crumbling when pinched". The soft material is squeezed through a 14-mesh sieve to make wet granules. The wet granules are dried at 50℃, 60℃ and 70℃ in time. The dried granules are sized by passing through a 14-mesh sieve and mixed evenly to obtain the final product.

[0167] Table 23 Investigation of drying temperature

[0168]

[0169] The overall score for drying temperature = moisture content × 25% + solubility × 25% + angle of repose × 25% + moldability × 25%

[0170] Based on the above temperature analysis, the particle score was optimal at 60℃, therefore the drying temperature of 60℃ was selected.

[0171] 3. Verification of Particle Molding Process

[0172] Weigh out 3 portions of 1 / 5 of the prescribed medicinal materials, extract, concentrate, and dry them separately according to the methods described above. Select the optimal mixed excipients previously screened, mix the dry extract powder and excipients according to the previously screened proportions, and select the previously screened ethanol solution to prepare a soft mass until it reaches a state where it "can be formed into a ball when grasped, but crumbles when squeezed." Extrude the soft mass through a 14-mesh sieve to form wet granules. Dry the wet granules promptly at 60°C, and then granulate them through a 14-mesh sieve. Mix well to obtain the final product. Validation is performed using the molding rate, angle of repose, solubility, and moisture content as indicators.

[0173] Table 24 Validation of Particle Molding Process

[0174]

[0175] Experimental results show that the particles prepared by the above-mentioned optimal molding process have a high overall score and good repeatability.

[0176] Example 4

[0177] This embodiment provides a method for preparing traditional Chinese medicine granules: Raw oyster shell and calcined mother-of-pearl are first decocted with 10 times the amount of water, soaked for 1.5 hours, and extracted three times, each time for 0.5 hours. The filtrate is filtered, and the three filtrates are combined. The decocted extract and the residue are combined with 9 other medicinal materials, soaked in 9 times the amount of water for 1.5 hours, and extracted twice, each time for 1 hour. The extract is filtered, centrifuged, and the supernatant is concentrated under reduced pressure at 50°C to obtain an extract (relative density 1.05 ~ 1.10 at 50°C). After drying under reduced pressure at 50°C, the extract is pulverized and passed through a 60-mesh sieve for later use.

[0178] The dry extract powder (33.33% of the prescription amount), lactose (44.09% of the prescription amount), mannitol (22.05% of the prescription amount), and steviol glycosides (0.53% of the prescription amount) are granulated with 85% ethanol, sieved, dried, granulated, and packaged (21.8 g / bag) to obtain the final product.

[0179] Example 5

[0180] Three batches of traditional Chinese medicine granules, each weighing 1000g, were prepared according to the optimal extraction and molding processes selected in Examples 2 and 3, respectively.

[0181] 1. Characteristics

[0182] This product consists of brownish-brown granules; it has an aromatic flavor and a slightly sweet taste.

[0183] 2. Identification

[0184] 2.1 Thin-layer chromatography identification of Isatis root

[0185] (1) Preparation of test solution: Take 3g of this product granules, grind them finely, add 20ml of 80% methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of methanol to dissolve the residue, and use it as test solution.

[0186] (2) Preparation of reference solution: Take (R,S)-Gauyichun reference standard and add methanol to prepare a solution containing 0.5mg per 1ml as the reference solution.

[0187] (3) Take another 1g of Isatis root reference material, add 20ml of 80% methanol, sonicate for 30 minutes, filter, evaporate the filtrate to dryness, add 1ml of methanol to dissolve the residue, and prepare a reference material solution.

[0188] (4) Negative control solution: Weigh the other ten medicinal materials that do not contain Isatis root according to the prescription ratio, and prepare a negative sample according to the molding process of wart removal granules. Take about 3g of the sample and prepare it in the same way as the test sample solution to obtain the negative control solution without Isatis root.

[0189] (5) Identification method and results: Perform thin-layer chromatography (General Rule 0502), take 5-10 μl of each of the above four solutions, and spot them separately on the same silica gel GF. 254 On thin-layer plates, petroleum ether (60–90 °C)-ethyl acetate (1:1) was used as the developing solvent. The plates were then removed, dried, and examined under ultraviolet light (254 nm). Figure 1 As shown, in the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatograms of the reference medicinal material and the reference standard.

[0190] 2.2 Thin-layer chromatography identification of Job's tears

[0191] (1) Preparation of test solution: Take 3g of this product granules, grind them finely, add 30ml of petroleum ether (60~90℃), sonicate for 30 minutes, filter, and take the filtrate as the test solution.

[0192] (2) Preparation of control extract solution: Take the coix seed oil control extract and dissolve it in petroleum ether (60-90℃) to prepare a coix seed oil control extract solution with a concentration of 2mg / ml.

[0193] (3) Take another 1g of Coix seed reference material, add 30ml of petroleum ether (60-90℃), sonicate for 30 minutes, filter, take the filtrate, and prepare the reference material solution.

[0194] (4) Negative control solution: Weigh the other ten medicinal materials that do not contain coix seed according to the prescription ratio, prepare a negative sample according to the molding process of wart removal granules, take about 3g of the sample, and prepare it in the same way as the test sample solution to obtain the negative control solution without coix seed.

[0195] (5) Identification method and results: Perform thin-layer chromatography (General Rule 0502). Apply 2 μl of each of the two solutions to the same silica gel G thin-layer plate. Develop using petroleum ether (60–90℃)-diethyl ether-glacial acetic acid (83:17:1) as the developing solvent. Remove the plate, air dry, spray with 5% vanillin-sulfuric acid solution, and heat at 105℃ until the spots are clearly visible. Figure 2 As shown, in the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference extract.

[0196] 2.3 Thin-layer chromatography identification of purslane

[0197] (1) Preparation of test solution: Take 3g of this product granules, add 20ml of water, add formic acid to adjust the pH value to 3-4, soak for 3 hours, filter, evaporate the filtrate to dryness, add 5ml of water to dissolve the residue, and use it as test solution.

[0198] (2) Preparation of control medicinal material solution: Take another 2g of purslane control medicinal material, add 20ml of water, add formic acid to adjust the pH value to 3-4, soak for 3 hours, filter, evaporate the filtrate to dryness, add 5ml of water to dissolve the residue, and use it as control medicinal material solution.

[0199] (3) Negative control solution: Weigh the other ten medicinal materials without purslane according to the prescription ratio, prepare the negative sample according to the molding process of wart removal granules, take about 3g of the sample, and prepare it in the same way as the test sample solution to obtain the negative control solution.

[0200] (4) Identification method and results: Perform thin-layer chromatography (General Rule 0502). Apply 1-2 μl of each of the two solutions to the same silica gel G thin-layer plate. Develop with a water-saturated n-butanol-glacial acetic acid-water (4:1:1) as the developing solvent. Remove the plate, air dry, spray with 0.2% ninhydrin ethanol solution, and heat at 110°C until the spots are clearly visible. Figure 3 As shown, in the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0201] 3. Inspection

[0202] 3.1 Particle size

[0203] Take 3 batches of samples and determine the particle size. The sum of those that cannot pass through sieve No. 1 and those that can pass through sieve No. 5 should not exceed 15%.

[0204] Table 25 Sample Particle Size Measurement Results

[0205]

[0206] 3.2 Moisture

[0207] Take 3 batches of samples and determine the moisture content; the moisture content must not exceed 8%.

[0208] Table 26 Results of moisture content determination in samples

[0209]

[0210] 3.3 Solubility

[0211] Take 10g of each of the three batches of samples, add 200mL of hot water in sequence, stir at a uniform speed for 5 minutes and observe immediately. The soluble particles should be completely dissolved or only slightly turbid.

[0212] Table 27 Results of Sample Solubility Test

[0213]

[0214] 4. Stability test

[0215] Three batches of wart-removing granules, in commercially available packaging, were placed in an environment with a temperature of 25℃±2℃ and a relative humidity of 60%±5%. The samples were taken out at 0, 3, 6, and 12 months to examine the properties, particle size, moisture content, and content of the samples at each time point.

[0216] Table 28 Sample stability test results

[0217]

Claims

1. A traditional Chinese medicine granule for treating flat warts, characterized in that, The granules comprise traditional Chinese medicine dry extract powder and excipients; the excipients comprise diluents, wetting agents and steviol glycosides; wherein the traditional Chinese medicine dry extract powder is obtained by extraction of the following components: 60 parts of Banlangen, 60 parts of Yiyiren, 60 parts of Mazi-xin, 30 parts of Zicao, 20 parts of Taoren, 20 parts of Honghua, 60 parts of Muzi, 20 parts of Xiangfu, 60 parts of Zhenzhumu, 20 parts of Fengfeng, and 60 parts of Shengmu-li.

2. The Chinese medicine granules for treating flat warts according to claim 1, characterized in that, The mass ratio of the dry extract powder and the excipients is 1:2-2.5, and the addition amount of steviol glycosides is 0.4-0.6% of the total mass of the excipients.

3. The Chinese medicine granules for treating flat warts according to claim 1, characterized in that, The diluents consist of lactose and mannitol; and the wetting agent is ethanol with a concentration of 85-95%.

4. The Chinese medicine granules for treating flat warts according to claim 1, characterized in that, The diluents consist of lactose and mannitol with a mass ratio of 2-1:

1.

5. A method for preparing the Chinese medicine granules for treating flat warts according to any one of claims 1-4, characterized in that, The method comprises the following steps: (1) soaking Shengmu-li and calcined Zhenzhumu in water, filtering the filtrate obtained after decocting to obtain a first decoction extract; (2) combining the first decoction extract, Shengmu-li residue, calcined Zhenzhumu residue and the other 9 medicinal materials, soaking in water, filtering the filtrate obtained after decocting, centrifuging, taking the supernatant, concentrating the supernatant under reduced pressure to obtain an extract, drying the extract under reduced pressure, and then crushing and sieving to obtain a dry extract powder; (3) mixing the dry extract powder and the excipients, and then granulating, drying and whole-granulating to obtain the traditional Chinese medicine granules.

6. The preparation method of the Chinese medicine granules for treating flat warts according to claim 5, characterized in that, In step (1), Shengmu-li and calcined Zhenzhumu are soaked in 8-10 times the amount of water of the total amount of medicinal materials for 1-2 h, and the filtrate obtained after decocting for 0.5-1.5 h is filtered, and the decoction is repeated 1-3 times, and the filtrate is combined to obtain the first decoction extract.

7. The method for preparing traditional Chinese medicine granules for treating flat warts according to claim 5, characterized in that, In step (2), the first decoction extract, Shengmu-li residue, calcined Zhenzhumu residue and the other 9 medicinal materials are combined, soaked in 8-10 times the amount of water of the total amount of the other 9 medicinal materials for 1.5-2.5 h, and the filtrate obtained after decocting for 0.75-2 h is filtered, and the decoction is repeated 1-3 times, and the supernatant is obtained by centrifuging, and the supernatant is concentrated under reduced pressure to obtain an extract, and the extract is dried under reduced pressure, and then crushed and sieved to obtain a dry extract powder.

8. The method for preparing traditional Chinese medicine granules for treating flat warts according to claim 5, characterized in that, In step (1), Shengmu-li and calcined Zhenzhumu are soaked in 8-10 times the amount of water of the total amount of medicinal materials for 1-2 h, and the filtrate obtained after decocting for 0.5-1.5 h is filtered, and the decoction is repeated 1-3 times, and the filtrate is combined to obtain the first decoction extract.

9. The method for preparing traditional Chinese medicine granules for treating flat warts according to claim 5, characterized in that, In step (3), the dry extract powder and the excipients are mixed, granulated, dried, and whole-granulated to obtain the traditional Chinese medicine granules.

10. The method for preparing traditional Chinese medicine granules for treating flat warts according to claim 9, characterized in that, The drying temperature is 50-70℃.

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