Nerve soothing and mind stabilizing pill and preparation method thereof

Abstract

The invention provides a nerve soothing and mind stabilizing pill and a preparation method thereof, and belongs to the technical field of traditional Chinese medicine preparations. The preparation method comprises the following steps: cleaning, infiltrating, cutting, drying, performing high-pressure steam sterilization on genuine medicinal materials such as poria cocos, poria with hostwood, ginseng, polygala tenuifolia, rhizoma acori graminei and the like in a formula of the mind-tranquilizing and mind-tranquilizing pill, mixing and crushing at low temperature to obtain fine powder, adding refined honey to prepare pills, and finally coating with cinnabar powder to obtain the mind-tranquilizing and mind-tranquilizing pill. The contents of polygala-ketone III, 3, 6 '-dimustard acyl sucrose, ginsenoside Rb1, beta-asarone, dehydrotumoic acid, pachymic acid and calcium salt in the prepared nerve-soothing and mind-stabilizing pill are high, the loss of index components in the pelleting process is small, and the dissolution time limit and microbial limit of the pill meet the requirements of Chinese Pharmacopoeia of 2025 edition. The preparation process of the mind-tranquilizing and mind-stabilizing pill is stable, and the prepared mind-tranquilizing and mind-stabilizing pill can effectively improve cognitive impairment, inflammation and oxidative stress injury caused by insomnia.

Description

Technical Field

[0001] The present invention relates to the field of traditional Chinese medicine preparations, and particularly to an Anshen Dingzhi Pills and its preparation method. Background Art

[0002] Pills are one of the oldest traditional dosage forms, with the advantages of convenient administration, stable and lasting efficacy. The classic famous prescription Anshen Dingzhi Pills (ADP) comes from "Medical Insights" Volume 4 • Insomnia by Cheng Guopeng in the Qing Dynasty, and has the effects of calming the mind and stabilizing the will, replenishing qi and suppressing fright, etc. It is a classic prescription for treating insomnia, anxiety, depression, palpitations, timidity and other diseases by medical experts of all dynasties. Anshen Dingzhi Pills are composed of Poria cocos, Poria cum Radix Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Dragon's Teeth. In the prescription, Poria cocos and Poria cum Radix Pini are combined to soothe the heart and calm the mind, and convey the heart qi to the kidney; Ginseng greatly replenishes qi, tonifying the qi of the spleen, lung, heart, and kidney. When qi is sufficient, blood is abundant and the mind is prosperous; Polygala tenuifolia enters the heart, kidney, and lung meridians, which can make the kidney qi rise to intersect with the heart yang to awaken the mind and improve intelligence and soothe the mind; Acorus tatarinowii opens the orifices to resolve phlegm, awakens the mind and improves intelligence, and promotes digestion by removing dampness; Dragon's Teeth and Cinnabar are used to suppress the hyperactive mind and calm the nerves; Honey is used to harmonize all the drugs, replenish the middle and moisten dryness, and enhance the synergistic efficacy of the whole prescription.

[0003] According to the "Requirements for Chinese Medicine Registration Classification and Declaration Materials" in China, Anshen Dingzhi Pills belong to Class 3.2 of other sources of ancient classic famous prescriptions for Chinese medicine registration. However, there are currently no reports on the preparation process of Anshen Dingzhi Pills. It is recorded in "Medical Insights" that Poria cum Radix Pini, Poria cocos, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Dragon's Teeth in Anshen Dingzhi Pills are pulverized in a ratio of 2:2:2:2:1:1. The specific preparation method is "the above drugs are ground into powder, refined honey is made into pills, and coated with cinnabar". However, the ratio of drug powder to honey, the drying temperature and time of the pills, and the safe dosage of the cinnabar coating are not specified in this traditional process; at the same time, how to meet the quality requirements such as the "Dissolution Time Limit" and "Microbial Limit" of pills in the 2025 edition of the "Chinese Pharmacopoeia" needs to be further studied. Summary of the Invention

[0004] The present invention aims to provide an Anshen Dingzhi Pills and its preparation method. The Anshen Dingzhi Pills prepared by the present invention have stable quality, high content of index components, meet the requirements of the "Dissolution Time Limit" and "Microbial Limit" of pills in the 2025 edition of the "Chinese Pharmacopoeia", and the dosage of cinnabar is lower than the requirements of the "Chinese Pharmacopoeia".

[0005] The present invention provides an Anshen Dingzhi Pills and the specific preparation steps are as follows: In order to achieve the above invention purpose, the present invention provides the following technical solutions: The present invention provides a preparation method of Anshen Dingzhi Pills, comprising the following steps: (1) Poria cocos, Poria cum Radix Pini, Ginseng, Polygala tenuifolia, and Acorus tatarinowii are respectively infiltrated, cut, and dried to obtain 五种 kinds of cut pieces; Dragon's Teeth is washed and then dried; (2) The Poria cocos, Poria cum Radice Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Draconis Cornu processed in step (1) are sterilized, dried, pulverized at low temperature, and mixed with refined honey to make pills, and then coated with cinnabar to obtain the Anshen Dingzhi Pills.

[0006] Preferably, the mass ratio of the Poria cocos, Poria cum Radice Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Draconis Cornu is Poria cocos:Poria cum Radice Pini:Ginseng:Polygala tenuifolia:Acorus tatarinowii:Draconis Cornu = 2 - 4:2 - 4:2 - 4:2 - 4:1 - 2:1 - 2.

[0007] Preferably, in step (1), the drying temperature of the Poria cocos, Poria cum Radice Pini, Ginseng, and Polygala tenuifolia after cutting is 60 - 70 °C, the drying temperature of the Acorus tatarinowii is 50 - 60 °C, and the drying temperature of the Draconis Cornu is 80 - 100 °C.

[0008] Preferably, in step (2), the sterilization temperature is 100 - 110 °C, and the time is 11 - 20 min.

[0009] Preferably, in step (2), the steps of low - temperature pulverization are as follows: The Poria cocos, Poria cum Radice Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Draconis Cornu are placed in a low - temperature pulverizer according to the ratio and pulverized into fine powder, and then sieved; among them, the weight of the fine powder passing through a 100 - mesh sieve should be greater than 98%.

[0010] Preferably, in step (2), the steps of refined - honey treatment are: heating the honey at 100 - 150 °C for 20 min - 30 min; the addition amount of the honey is 20 g - 60 g of honey per 100 g of the mixed powder.

[0011] The present invention also provides Anshen Dingzhi Pills prepared by the preparation method of the above - mentioned Anshen Dingzhi Pills.

[0012] The beneficial effects of the present invention compared with the prior art are as follows: (1) The present invention combines traditional Chinese medicine pharmaceutical experience with modern drug quality control standards, and provides a preparation method of the classic famous prescription Anshen Dingzhi Pills. After washing, moistening, cutting, sterilizing, drying, and pulverizing the genuine medicinal materials Poria cocos, Poria cum Radice Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Draconis Cornu, fine powder of the drug is obtained, and then refined honey is added to make small pills, and finally the Anshen Dingzhi Pills are prepared by coating with cinnabar. The Anshen Dingzhi Pills prepared by the present invention have stable quality, definite curative effect, and clear action mechanism. The contents of its index components polygalaxanthone III, 3,6'-di - sinapoyl - sucrose, ginsenoside Rb1, β - asarone, dehydrotumulosic acid, pachymic acid, and the content of calcium salts are stable, and it has a good dissolution effect. At the same time, the Anshen Dingzhi Pills prepared by the present invention fully meet the microbial limit requirements for traditional Chinese medicine pills in the 2025 edition of the Chinese Pharmacopoeia, providing a solid scientific basis for the new drug development and clinical popularization and application of the Anshen Dingzhi Pills.

[0013] (2) The high-pressure steam sterilization method adopted by the present invention has good sterilization effect and less loss of active ingredients. At the same time, low-temperature pulverization effectively prevents the loss of medicinal components, and the proportion of fine powder passing through a 100-mesh sieve is as high as 98%, effectively ensuring the quality uniformity and drug dissolution rate of Anshen Dingzhi Pills.

[0014] (3) The optimized ratio of medicinal powder to refined honey in the present invention can effectively ensure the stability of pill shape and resistance to moisture absorption, and can maintain high production efficiency and stability in industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required to be used in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0016] Figure 1 It is the process route diagram for the preparation of Anshen Dingzhi Pills of the present invention; Figure 2 It is the appearance character diagram of the Anshen Dingzhi Pills prepared in Examples 1 to 3 of the present invention; Figure 3 It is the chromatogram for the determination of the contents of polygalaxanthone III, 3,6'-di-O-sinapoyl-sucrose, ginsenoside Rb1, β-asarone, dehydrotumulosic acid and pachymic acid in the Anshen Dingzhi Pills prepared in Example 1 of Test Example 2 of the present invention; where a is polygalaxanthone III, b is 3,6'-di-O-sinapoyl-sucrose, c is ginsenoside Rb1, d is β-asarone, e is dehydrotumulosic acid, and f is pachymic acid; Figure 4 It is the chromatogram for the determination of the contents of polygalaxanthone III, 3,6'-di-O-sinapoyl-sucrose, ginsenoside Rb1, β-asarone, dehydrotumulosic acid and pachymic acid in the Anshen Dingzhi Pills prepared in Comparative Example 1 of Test Example 2 of the present invention; where a is polygalaxanthone III, b is 3,6'-di-O-sinapoyl-sucrose, c is ginsenoside Rb1, d is β-asarone, e is dehydrotumulosic acid, and f is pachymic acid; Figure 5 It is the chromatogram for the determination of the contents of polygalaxanthone III, 3,6'-di-O-sinapoyl-sucrose, ginsenoside Rb1, β-asarone, dehydrotumulosic acid and pachymic acid in the Anshen Dingzhi Pills prepared in Comparative Example 2 of Test Example 2 of the present invention; where a is polygalaxanthone III, b is 3,6'-di-O-sinapoyl-sucrose, c is ginsenoside Rb1, d is β-asarone, e is dehydrotumulosic acid, and f is pachymic acid; Figure 6This is a titration chart for the determination of calcium salt content in An Shen Ding Zhi Wan prepared in Example 1 of Experimental Example 3 of the present invention; where a is the titration color after adding methyl red indicator solution, b is the titration color after adding 10% potassium hydroxide, c is the titration color after adding calcein, and d is the titration color after adding 0.05069 mol / L disodium ethylenediaminetetraacetate. Figure 7 This is a titration chart for the determination of calcium salt content in An Shen Ding Zhi Wan prepared in Example 1 of Experimental Example 3 of the present invention; where a is the titration color after adding methyl red indicator solution, b is the titration color after adding 10% potassium hydroxide, c is the titration color after adding calcein, and d is the titration color after adding 0.05069 mol / L disodium ethylenediaminetetraacetate. Figure 8 This is a titration chart for the determination of calcium salt content in An Shen Ding Zhi Wan prepared in Comparative Example 2 of Experimental Example 3 of the present invention; where a is the titration color after adding methyl red indicator solution, b is the titration color after adding 10% potassium hydroxide, c is the titration color after adding calcein, and d is the titration color after adding 0.05069 mol / L disodium ethylenediaminetetraacetate. Figure 9 This is a titration chart for the determination of mercuric sulfide content in An Shen Ding Zhi Wan prepared in Example 1 of Experimental Example 4 of the present invention; where a is the titration color after adding 1% potassium permanganate solution, b is the titration color after adding 2% ferrous sulfate solution, and c is the titration color after adding ferric ammonium sulfate indicator solution and 0.1 mol / L ammonium thiocyanate titrant. Figure 10 This is a titration chart for the determination of mercuric sulfide content in An Shen Ding Zhi Wan prepared in Example 2 of Experimental Example 4 of the present invention; where a is the titration color after adding 1% potassium permanganate solution, b is the titration color after adding 2% ferrous sulfate solution, and c is the titration color after adding ferric ammonium sulfate indicator solution and 0.1 mol / L ammonium thiocyanate titrant. Figure 11 This is a titration chart for the determination of mercuric sulfide content in An Shen Ding Zhi Wan prepared in Comparative Example 2 of Experimental Example 4 of the present invention; where a is the titration color after adding 1% potassium permanganate solution, b is the titration color after adding 2% ferrous sulfate solution, and c is the titration color after adding ferric ammonium sulfate indicator solution and 0.1 mol / L ammonium thiocyanate titrant. Figure 12 The figure shows the effect of An Shen Ding Zhi Wan (ADP) on the localization and navigation ability and spatial exploration ability of sleep-deprived rats in the Morris water maze in Experiment Example 6 of this invention, where (A represents localization and navigation ability; B represents spatial exploration ability). Figure 13 The effect of ADP on pathological damage in the CA1 region of sleep-deprived rats in Experiment Example 6 of this invention (×20). Figure 14This is a bar graph showing the effects of ADP on the levels of inflammatory factors IL-1β, IL-6, and TNF-α in the serum of sleep deprivation model rats in Experimental Example 6 of the present invention. Figure 15 This is a bar graph showing the effects of ADP on the levels of oxidative stress SOD, MDA, and GSH in the brain tissue of sleep deprivation model rats in Experimental Example 6 of the present invention. Detailed implementation manners

[0017] Now, the various exemplary implementation manners of the present invention will be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0018] It should be understood that the terms used in the present invention are only for describing specific implementation manners and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0019] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0020] Without departing from the scope or spirit of the present invention, various improvements and changes can be made to the specific implementation manners of the present invention's specification, which are obvious to those skilled in the art. Other implementation manners obtained from the present invention's specification are obvious to those skilled in the art. The present invention's specification and examples are merely exemplary.

[0021] Regarding the use of "comprising", "including", "having", "containing", etc. in this article, they are all open-ended terms, meaning including but not limited to.

[0022] The present invention provides a preparation method for Anshen Dingzhi Pills, and the specific steps are as follows: (1) Wash, moisten, cut, and dry the authentic Chinese medicinal materials Poria, Poria Pararadicis, Ginseng Radix, Polygalae Radix, and Acori Tatarinowii Rhizoma respectively to obtain 6 kinds of cut slices; wash and dry the Draconis Os; the drying temperature of Poria, Poria Pararadicis, Ginseng Radix, and Polygalae Radix after cutting is 60 - 70 °C, the drying temperature of Acori Tatarinowii Rhizoma is 50 - 60 °C, and the drying temperature of Draconis Os is 80 - 100 °C.

[0023] (2) Sterilize the 6 kinds of cut slices processed in step (1) with high-pressure steam at 100 - 110 °C for 5 - 20 min, and then transfer them to the clean area for drying; the drying temperature of Poria, Poria Pararadicis, Ginseng Radix, and Polygalae Radix after cutting is 60 - 70 °C, the drying temperature of Acori Tatarinowii Rhizoma is 50 - 60 °C, and the drying temperature of Draconis Os is 80 - 100 °C.

[0024] (3) Put the dried and sterilized cut slices into a pulverizer according to the ratio of Poria Pararadicis:Poria:Ginseng Radix:Polygalae Radix:Acori Tatarinowii Rhizoma:Draconis Os of 2:2:2:2:1:1, and mix and pulverize at -20 - -10 °C for 11 - 15 min; sieve the obtained fine powder, and require that the weight of the fine powder passing through a 100-mesh sieve is not less than 98% to ensure the effective dissolution rate and uniformity of the pills.

[0025] (4) Mix the above-mentioned mixed medicinal powder and refined honey evenly according to the ratio of 1:0.2 - 0.6 to make soft materials, and use the extrusion-spheronization method to make pills, where the feeding speed for pill making is 0.5 m / min and the extrusion-spheronization speed is 80 r / min. Finally, coat and polish with the ratio of 1.0 - 1.5 g of fine cinnabar powder for every 100 g of medicinal powder to make small pills with a diameter of 6 - 8 mm and a pill weight of about 0.2 - 0.4 g.

[0026] (5) Finally, dry at a low temperature of 30 - 35 °C for 14 - 20 hours to obtain Anshen Dingzhi Pills with a moisture content of 7 - 8.5% and stable index component content.

[0027] Example 1 Example 1 of the present invention provides a preparation method of Anshen Dingzhi Pills, and the specific steps are as follows: (1) Wash, moisten and soften, and cut the authentic Chinese medicinal materials Ginseng Radix, Polygalae Radix, Poria, Poria Pararadicis, and Acori Tatarinowii Rhizoma respectively, and then dry Poria, Poria Pararadicis, Ginseng Radix, and Polygalae Radix at 65 °C for 4 h, dry Acori Tatarinowii Rhizoma at 55 °C for 6 h, and dry Draconis Os at 90 °C for 2 h to obtain Ginseng Radix cut slices, Polygalae Radix cut slices, Poria cut slices, Poria Pararadicis cut slices, Acori Tatarinowii Rhizoma cut slices, and Draconis Os cut slices; (2) Sterilize the Ginseng Radix cut slices, Polygalae Radix cut slices, Poria cut slices, Poria Pararadicis cut slices, Acori Tatarinowii Rhizoma cut slices, and Draconis Os cut slices processed in step (1) at 100 °C for 5 min and enter the clean area, and then dry in the clean area oven until the moisture content is less than 6%; (3) Put the sterilized and dried Chinese herbal pieces into a pulverizer according to the ratio of Poria cum Radix Pini: Poria: Ginseng: Polygala tenuifolia: Acorus tatarinowii: Draconis denticula 2:2:2:2:1:1, and pulverize them at a low temperature of -20°C for 12 minutes, then sieve. The weight of the fine powder passing through a 100-mesh sieve is 99.2%; (4) Heat the honey at 150°C for 20 min to obtain refined honey. Mix the pulverized and evenly mixed medicinal powder with the refined honey in a ratio of 1:0.4 to make a soft material, and use the extrusion and spheronization method to make pills. The feeding speed for pill making is 0.5 m / min, and the extrusion and spheronization speed is 80 r / min.

[0028] (5) Coat and polish the small pills obtained in step (4) with fine cinnabar powder to make Anshen Dingzhi Pills with a diameter of 6 - 8 mm and a pill weight of about 0.3 g, and dry them at 35°C for 18 hours to obtain Anshen Dingzhi Pills.

[0029] Example 2 The present invention provides a preparation method of Anshen Dingzhi Pills in Example 2, and the specific steps are as follows: (1) Wash, soak and soften, and cut the five Chinese herbal medicines of ginseng, polygala tenuifolia, poria, Poria cum Radix Pini, and acorus tatarinowii respectively. Then dry poria, Poria cum Radix Pini, ginseng, and polygala tenuifolia at 70°C for 3 h, dry acorus tatarinowii at 55°C for 6 h, and dry draconis denticula at 90°C for 2 h to obtain ginseng pieces, polygala tenuifolia pieces, poria pieces, Poria cum Radix Pini pieces, acorus tatarinowii pieces, and draconis denticula pieces; (2) Subject the ginseng pieces, polygala tenuifolia pieces, poria pieces, Poria cum Radix Pini pieces, acorus tatarinowii pieces, and draconis denticula pieces processed in step (1) to high-pressure sterilization at 105°C for 5 min and then enter the clean area, and then dry them in the clean area oven until the moisture content is less than 6%; (3) Put the sterilized and dried Chinese herbal pieces into a pulverizer according to the ratio of Poria cum Radix Pini: Poria: Ginseng: Polygala tenuifolia: Acorus tatarinowii: Draconis denticula 2:2:2:2:1:1, and pulverize them at a low temperature of -15°C for 12 minutes, then sieve. The weight of the fine powder passing through a 100-mesh sieve is 99.5%; (4) Heat the honey at 125°C for 25 min to obtain refined honey. Mix the pulverized and evenly mixed medicinal powder with the refined honey in a ratio of 1:0.5 to make a soft material, and use the extrusion and spheronization method to make pills. The feeding speed for pill making is 0.5 m / min, and the extrusion and spheronization speed is 80 r / min.

[0030] (5) Coat and polish the small pills obtained in step (4) with fine cinnabar powder to make Anshen Dingzhi Pills with a diameter of 6 - 8 mm and a pill weight of about 0.4 g, and dry them at 38°C for 17 hours to obtain Anshen Dingzhi Pills.

[0031] Example 3 The present invention provides a preparation method of Anshen Dingzhi Pills in Example 1, and the specific steps are as follows: (1)依次清洗、浸润软化并切制人参、远志、茯苓、茯神、石菖蒲这五味药材,然后将茯苓、茯神、人参、远志在65℃干燥4小时,石菖蒲在55℃干燥6小时,龙齿在100℃干燥1小时,从而得到人参饮片、远志饮片、茯苓饮片、茯神饮片、石菖蒲饮片和龙齿饮片; (2)将步骤(1)处理后的人参饮片、远志饮片、茯苓饮片、茯神饮片、石菖蒲饮片、龙齿饮片在110℃下进行高压灭菌5分钟,之后进入洁净区,并在洁净区烘箱中干燥至水分小于6%; (3)把灭菌干燥后的饮片按照茯神:茯苓:人参:远志:石菖蒲:龙齿比例2:2:2:2:1:1放入粉碎机,在-10℃低温混合粉碎15分钟,然后过筛,通过100目筛的细粉重量为99.8%; (4)将蜂蜜在100℃加热30分钟得到炼蜜。把粉碎混合均匀的药粉与炼蜜按照1: 0.5的比例混合均匀制成软材,采用挤出滚圆法制丸,其中制丸送料速度为0.5 m / min,挤出滚圆速度为80 r / min。

[0032] (5)用朱砂细粉对步骤(4)所得小丸进行包衣并抛光,制成直径为6 - 8 mm、丸重约0.2 g的安神定志丸,并在35℃干燥18小时制得安神定志丸。

[0033] Comparative Example 1 In Comparative Example 1 of the present invention, Anshen Dingzhi Pills were prepared using the same mass ratio and method as in Example 1. The difference from Example 1 is that step (2) of Comparative Example 1 does not include the high-temperature sterilization step in Example 1.

[0034] Comparative Example 2 In Comparative Example 2 of the present invention, Anshen Dingzhi Pills were prepared using the same mass ratio and method as in Example 1. The difference from Example 1 is that the pulverization time in step (3) of Comparative Example 2 is 4 minutes.

[0035] Comparative Example 3 In Comparative Example 3 of the present invention, commercially available Anshen Dingzhi Pills were used. The difference from Example 1 is that Comparative Example 3 consists of Poria cum Radix Pini, Poria, Polygala tenuifolia, Acorus tatarinowii, and Panax ginseng.

[0036] Test Example 1 In Test Example 1 of the present invention, the dissolution time limit of the Anshen Dingzhi Pills prepared in Example 1, Example 2, Example 3, and Comparative Example 3 was determined. The specific determination method is as follows: According to the method described in Section IV of the Chinese Pharmacopoeia 2020, under the Disintegration Test Method (General Rule 0921) for tablets, the test was carried out with a baffle. It is stipulated that the hanging basket is suspended on the bracket through the upper stainless steel shaft, immersed in a 1000 mL beaker, and the position of the hanging basket is adjusted so that when it drops to the lowest point, the sieve is 25 mm from the bottom of the beaker. The beaker contains water at a temperature of 37°C ± 1°C. The water level is adjusted so that when the hanging basket rises to the highest point, the sieve is 15 mm below the water surface, and the top of the hanging basket must not be immersed in the solution. All should pass through the sieve within the specified 1 hour. If there are fine granular substances that do not pass through the sieve but have softened and have no hard core, it can be considered in compliance with the regulations. The disintegration test results of sixteen batches of the reference samples of Anshen Dingzhi Pills are shown in Table 1: Table 1 Dissolution Time Limit of Anshen Dingzhi Pills

[0037] As can be seen from Table 1, the dissolution time limits (disintegration time limits) of the Anshen Dingzhi Pills prepared in Examples 1 to 3 all comply with the regulations of the Chinese Pharmacopoeia 2025 Edition.

[0038] Test Example 2 In Test Example 2 of the present invention, the contents of the index components of yuanzhi ketone III, 3,6'-di-sinapoyl sucrose, ginsenoside Rb1, β-asarone, dehydrotumulosic acid and pachymic acid in the Anshen Dingzhi Pills prepared in Examples 1, Example 2, Example 3 and Comparative Example 3 were determined. The specific determination methods for each index component are as follows: Determined according to the high performance liquid chromatography method in General Rule 0512 of the Chinese Pharmacopoeia 2020 Edition. Among them, 0.1% phosphoric acid aqueous solution is used as mobile phase A, and acetonitrile is used as mobile phase B for gradient elution; the column temperature is 35°C; the flow rate is 1.0 mL per minute, and the detection wavelength is 203 nm.

[0039] Table 2 Mobile Phase Elution Gradient

[0040] Preparation of the reference solution: Weigh the reference substances of yuanzhi ketone III, 3,6'-di-sinapoyl sucrose, ginsenoside Rb1, β-asarone, dehydrotumulosic acid and pachymic acid, add methanol solution to dissolve and make up the volume to prepare a mixed reference solution with concentrations of 0.07 mg / mL, 0.25 mg / mL, 0.10 mg / mL, 0.10 mg / mL, 0.03 mg / mL and 0.04 mg / mL respectively.

[0041] Preparation of the test solution: Weigh about 5g of Anshen Dingzhi Pill reference sample, accurately add 100mL of water-saturated n-butanol, seal tightly, let stand for 30min, sonicate (power 250W, frequency 50kHz) for 30min, filter, discard the initial filtrate, accurately measure 25mL of the subsequent filtrate, evaporate to dryness in an evaporating dish, dissolve the residue in methanol and transfer to a 5mL volumetric flask, dilute with methanol to the mark, shake well, filter, and take the subsequent filtrate to obtain the test solution.

[0042] Determination method: Accurately pipette 10 µL each of the reference solution and the test solution into the liquid chromatograph, and determine the results. The results are as follows: Figures 3-5 As shown in Tables 3 and 4.

[0043] Table 3. Content of key components of Anshen Dingzhi Pills in three different formulation processes

[0044] Table 4. Change rate (%) of index components of Anshen Dingzhi Pill in three preparation processes

[0045] Depend on Figures 3-6 As shown in Tables 3 and 4, the An Shen Ding Zhi Wan prepared in Examples 1-3 of this invention has high contents of the indicative components such as polygalone III, 3,6'-disinoyl sucrose, ginsenoside Rb1, β-asarone, dehydrotomolic acid and pamoic acid, which can effectively ensure the clinical therapeutic effect of An Shen Ding Zhi Wan.

[0046] Experimental Example 3 In Experiment 1 of this invention, the calcium salt content in the An Shen Ding Zhi Wan prepared in Example 1, Comparative Example 1, and Comparative Example 2 was determined. The specific determination method is as follows: Preparation of the test solution: Accurately weigh about 3g of this product, place it in an Erlenmeyer flask, add 125mL of dilute hydrochloric acid, sonicate to dissolve, and add 500mL of water to prepare the test solution.

[0047] Titration method: Accurately measure 105 mL of the test solution, add 1 drop of methyl red indicator, and add 10% potassium hydroxide solution until the solution turns yellow. Then add a small amount of calcein indicator and titrate with disodium ethylenediaminetetraacetate titrant (0.05069 mol / L) until the yellow-green fluorescence of the solution disappears and turns orange. Each 1 mL of disodium ethylenediaminetetraacetate titrant (0.050 mol / L) is equivalent to 4.069 mg of calcium carbonate. The calcium salt content of this product is calculated based on calcium carbonate, and the results are as follows: Figures 6-8 As shown in Table 5.

[0048] Table 5 Calcium salt content in Anshen Dingzhi Pills

[0049] Depend on Figures 6-8 As can be seen from Table 5, the Anshen Dingzhi Pills prepared by the present invention have a high calcium salt content and good effects.

[0050] Test Example 4 In Test Example 1 of the present invention, the content of mercury sulfide in the Anshen Dingzhi Pills prepared in Examples 1, 2, and 3 was determined. The specific method is as follows: Titration method: Take about 1.0 g of this product, accurately weigh it, place it in a conical flask, add 30 mL of sulfuric acid and 4.5 g of potassium nitrate. Place the conical flask on an electric furnace and heat it with an open flame for digestion. Heat until the solution turns light yellowish-white, cool it to room temperature, add 50 mL of water, dropwise add 1% potassium permanganate solution until it shows a pink color, then dropwise add 2% ferrous sulfate solution until the red color disappears, and then add 2 mL of ammonium ferric sulfate indicator solution. Titrate with ammonium thiocyanate titrant (0.1 mol / L) until the solution shows a light orange-red color. Each 1 mL of ammonium thiocyanate titrant (0.1 mol / L) is equivalent to 11.63 mg of mercury sulfide (HgS). The cinnabar content of this product is calculated based on mercury sulfide (HgS). The results are as Figures 9-11 shown in Table 6.

[0051] Table 6 Mercury sulfide content in Anshen Dingzhi Pills

[0052] From Figures 9-11 and Table 6, it can be seen that the content of mercury sulfide in the Anshen Dingzhi Pills prepared in Examples 1 - 3 of the present invention is within the cinnabar dosage range in the Chinese Pharmacopoeia 2025 Edition.

[0053] Test Example 5 According to the microbial limit test method (General Rule 1105), the test for controlled microorganisms (General Rule 1106) and the microbial limit standards for non-sterile drugs (General Rule 1107) in the Chinese Pharmacopoeia 2025 Edition, the microorganisms in the six kinds of prepared slices of Poria cocos, Poria cum Radix Pini, Ginseng Radix, Polygala tenuifolia, Acori Tatarinowii Rhizoma, and Draconis Odontolithus after sterilization and drying in Examples 1 - 3 of the present invention and the finally prepared Anshen Dingzhi Pills were detected. The results are shown in Tables 7 - 13: Table 7 Detection results of microbial limits of Poria cocos prepared slices

[0054] Conclusion: The microbial limits of Poria cocos prepared slices after sterilization and drying meet the requirements of the Chinese Pharmacopoeia 2025 Edition.

[0055] Table 8 Detection results of microbial limits of Poria cum Radix Pini prepared slices

[0056] Conclusion: The microbial limits of Poria cum Radix Pini prepared slices after sterilization and drying meet the requirements of the Chinese Pharmacopoeia 2025 Edition.

[0057] Table 9 Detection results of microbial limits of Ginseng Radix prepared slices

[0058] Conclusion: The microbial limits of sterilized and dried ginseng slices meet the requirements of the 2025 edition of the Chinese Pharmacopoeia.

[0059] Table 10 Results of Microbial Limit Tests on Polygala tenuifolia Root Slices

[0060] Conclusion: The microbial limits of sterilized and dried Polygala tenuifolia slices meet the requirements of the 2025 edition of the Chinese Pharmacopoeia.

[0061] Table 11 Results of Microbial Limit Tests on Acorus tatarinowii Slices

[0062] Conclusion: The microbial limits of sterilized and dried Acorus tatarinowii slices meet the requirements of the 2025 edition of the Chinese Pharmacopoeia.

[0063] Table 12 Results of Microbial Limit Tests on Dragon Tooth Slices

[0064] Conclusion: The microbial limits of sterilized and dried dragon teeth slices meet the requirements of the 2025 edition of the Chinese Pharmacopoeia.

[0065] Table 13 Results of Microbial Limit Tests for Anshen Dingzhi Pills

[0066] Conclusion: The microbial limits of the An Shen Ding Zhi Wan prepared in Examples 1-3 of this invention all meet the requirements of the 2025 edition of the Chinese Pharmacopoeia.

[0067] Experimental Example 6 Example 6 of this invention investigated the efficacy of the An Shen Ding Zhi Wan (ADP) prepared in Example 1 on a sleep deprivation model rat (Model). The specific steps are as follows: Normal rats were used as a blank control group. Sleep deprivation model rats were randomly divided into four groups: a model group (Model), a low-dose Anshen Dingzhi Wan group (ADP-L 0.75 g / kg / d), a medium-dose Anshen Dingzhi Wan group (ADP-M 1.5 g / kg / d), and a high-dose Anshen Dingzhi Wan group (ADP-H 3.0 g / kg / d). The low, medium, and high doses of Anshen Dingzhi Wan were administered by gavage at 0.5, 1, and 2 times the clinical adult equivalent dose, respectively. The control group and the model group were administered an equal volume of physiological saline by gavage. After 7 days of gavage, the sleep deprivation model rats' navigation and spatial exploration abilities in the Morris water maze, as well as the effects on the rats, were observed. The results are as follows: Figures 12-15 As shown.

[0068] Figure 12 The results showed that the number of times rats crossed the platform and the time spent in the target quadrant were significantly reduced in the model group (P<0.01), while the low, medium and high dose groups of Anshen Dingzhi Pill (ADP-L, ADP-M, ADP-H) and the positive control drug melatonin group (Melatonin, 0.10 g / kg) all increased the number of times rats crossed the platform and the time spent in the target quadrant, and shortened the escape latency of rats. This indicates that Anshen Dingzhi Pill prepared in this invention can effectively improve the cognitive level of sleep deprivation model rats.

[0069] Figure 13 The control group showed that neurons in the CA1 region of the hippocampus of rats were well-formed, tightly packed, clearly structured, with uniformly stained nuclei, moderate intercellular spaces, abundant neurons, and no obvious lesions. In contrast, the model group showed disordered and sparse neuronal arrangement, irregular morphology, reduced number, and significant nuclear condensation in the CA1 region of the hippocampus of rats treated with low, medium, and high doses of An Shen Ding Zhi Wan (ADP-L, ADP-M, ADP-H) and the melatonin group showed more orderly cell arrangement and a significantly increased number of neurons in the CA1 region of the hippocampus. This indicates that the ADP prepared in this invention can effectively alleviate hippocampal neuronal damage in sleep-deprived rats.

[0070] Figure 14 The concentrations of inflammatory factors IL-1β, IL-6, and TNF-α in the serum of model rats were significantly higher than those in the control group (P<0.01). Compared with the model, the levels of IL-1β, IL-6, and TNF-α in rats in the low, medium, and high dose groups of An Shen Ding Zhi Wan and the melatonin group were significantly lower (P<0.05, 0.01, respectively). Moreover, the anti-inflammatory effect of ADP showed a dose-dependent relationship, indicating that the anti-inflammatory effect of the ADP prepared in this invention was further enhanced with increasing drug dosage.

[0071] Figure 15Compared with the normal group, the expression of SOD and GSH in the brain tissue of rats in the model group was significantly decreased (P<0.01), and the concentration of MDA was significantly increased (P<0.01), indicating that sleep deprivation caused significant oxidative stress damage in the rat brain tissue. Compared with the model group, the concentrations of SOD and GSH in the brain tissue of rats in the low, medium, and high dose groups of Anshen Dingzhi Pill and the melatonin group were significantly increased (P<0.01), and the MDA level was significantly decreased (P<0.01). The results show that ADP can significantly improve the level of oxidative stress in the brain tissue of sleep-deprived rats, and this improvement effect is significantly dose-dependent. With the increase of drug dose, the levels of SOD and GSH are higher, and the content of MDA is lower. The results indicate that Anshen Dingzhi Pill prepared in this invention can effectively improve oxidative stress damage in the brain tissue of sleep-deprived rats, providing a scientific reference for the development and clinical application of Anshen Dingzhi Pill as a new anti-insomnia drug.

[0072] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for preparing a calming and stabilizing pill, characterized in that, It includes the following steps: (1) Soak, cut and dry Poria cocos, Poria cum Radix Pini, Ginseng, Polygala tenuifolia, and Acorus tatarinowii respectively to obtain 5 kinds of cut crude drugs; wash and dry Dragon's Teeth; (2) Sterilize, dry by baking, pulverize at low temperature, add refined honey to make pills, and coat with cinnabar on the Poria cocos, Poria cum Radix Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Dragon's Teeth processed in step (1) to obtain Anshen Dingzhi Pills.

2. The preparation method of An Shen Ding Zhi Wan according to claim 1, characterized in that, The mass ratio of the Poria cocos, Poria cum Radix Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Dragon's Teeth is Poria cocos:Poria cum Radix Pini:Ginseng:Polygala tenuifolia:Acorus tatarinowii:Dragon's Teeth = 2 - 4:2 - 4:2 - 4:2 - 4:1 - 2:1 - 2.

3. The preparation method of Anshen Dingzhi Pill according to claim 1, characterized in that, In step (1), the drying temperature of the Poria cocos, Poria cum Radix Pini, Ginseng, and Polygala tenuifolia after cutting is 60 - 70°C, the drying temperature of the Acorus tatarinowii is 50 - 60°C, and the drying temperature of the Dragon's Teeth is 80 - 100°C.

4. The preparation method of An Shen Ding Zhi Wan according to claim 1, characterized in that, In step (2), the sterilization temperature is 100 - 110°C and the time is 11 - 20 min.

5. The preparation method of An Shen Ding Zhi Wan according to claim 1, characterized in that, In step (2), the step of pulverizing at low temperature is as follows: Place the Poria cocos, Poria cum Radix Pini, Ginseng, Polygala tenuifolia, Acorus tatarinowii, and Dragon's Teeth in proportion in a low-temperature pulverizer, pulverize them into fine powder, and sieve; among them, the weight of the fine powder passing through a 100-mesh sieve should be greater than 98%.

6. The preparation method of An Shen Ding Zhi Wan according to claim 1, characterized in that, In step (2), the step of processing with refined honey is: heat the honey at 100 - 150°C for 20 min - 30 min; the addition amount of the honey is 20 g - 60 g of honey per 100 g of the mixed powder.

7. Anshen Dingzhi Pills prepared by the preparation method of the Anshen Dingzhi Pills according to any one of claims 1 - 5.

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