Preparation method and application of traumatic injury seven-centimeter preparation based on raw material granularity control

Abstract

The invention relates to a preparation method and application of a traumatic injury Qicentimeter preparation based on raw material granularity control, and the traumatic injury Qicentimeter preparation is prepared from the following medicinal raw materials in parts by mass: 0.8 part of artificial musk, 8 parts of pseudo-ginseng, 16 parts of dragon's blood, 48 parts of safflower, 40 parts of cinnabar, 80 parts of wine-processed angelica sinensis, 32 parts of vinegar-processed myrrh, 1.6 parts of borneol, 32 parts of vinegar-processed frankincense and 40 parts of catechu. The preparation method of the pseudo-ginseng preparation comprises the following steps: S1, raw material pretreatment and crushing: crushing the medicinal raw materials, and controlling the crushing granularity of pseudo-ginseng to be 80-120 meshes; the crushing granularity of the dragon's blood is controlled between 60 meshes and 100 meshes; s2, total mixing and preparation: mixing the crushed medicinal raw materials to obtain a mixture, and preparing the mixture into the traumatic injury seven-centimeter preparation. On the basis of the specific preparation, under the condition of not adopting a complex process, the bioavailability and the industrial preparation feasibility are synchronously improved only by accurately controlling the specific crushing particle size range of the key medicines.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine, and particularly relates to a preparation method and application of a Qiqi pill based on raw material particle size control. BACKGROUND

[0002] The Qiqi pill is derived from the Qiqi powder, a holy medicine for orthopedics, which was first recorded in the Lianfang Jixi (Collection of Good Prescriptions) compiled by Xie Yuanqing, a medical practitioner in the Qing Dynasty. It has the effects of resolving stasis, reducing swelling, stopping pain, and stopping bleeding, and is used for treating injuries from falls and blows, blood stasis pain, and external injuries with bleeding. It has good effects on various fractures, soft tissue injuries, and external injuries with bleeding. In modern preparations, the Qiqi pill is mainly made from angelica sinensis, safflower, frankincense, myrrh, dragon's blood, panax notoginseng, musk, borneol, cinnabar, and catechu through processes such as crushing, granulating, and tabletting.

[0003] At present, there are mainly two representative technologies for the production process of the Qiqi pill. The patent document with the literature number CN1840052C discloses a preparation method of the Qiqi pill. The cinnabar is crushed into 120-200 mesh fine powder, and the rest of the medicinal materials are crushed into 60-80 mesh fine powder. After mixing, granulation, drying, and tabletting are performed. Although this process is simple and easy to industrialize, the crushing particle size of the medicinal materials is relatively coarse (especially for the main drugs such as panax notoginseng and dragon's blood), which leads to slow dissolution of effective components, low bioavailability, and affects the full play of the drug effect.

[0004] The patent document with the publication number CN113633708A discloses a preparation method of a stable Qiqi composition tablet. In view of the stability problems of volatile and degradable components such as dragon's blood and borneol, hydrogenated castor oil is used for closed protection, and the D90 particle size of the drug mixture is controlled to be 40-80 μm (about equivalent to 180-400 mesh) to improve the tablet hardness and component stability. However, this process requires extremely fine crushing of the medicinal materials, which is high in energy consumption and equipment loss during production, difficult to realize large-scale continuous production, and the fine powder is easy to cause material adsorption, hardening, poor flowability, further affecting the stability and yield of the granulation and tabletting processes. In addition, the content of resin raw materials is large in the production of the Qiqi pill by the existing process, which is easy to adsorb moisture, leading to hardening and moisture absorption, and further causing molding difficulty.

[0005] In summary, the existing technology has the following outstanding technical problems: Low bioavailability: due to the relatively coarse crushing particle size of some medicinal materials, the effective components are not fully dissolved, which limits the drug effect; Complex process and high energy consumption: excessive pursuit of superfine crushing for component stability leads to high production cost and difficulty in industrialization; Poor material flowability and poor compressibility: fine powder is easy to absorb moisture and harden, affecting the stability of the preparation process and the quality of the tablets.

[0006] Therefore, there is an urgent need in the art for a preparation method of Dieda Qili preparation which can ensure the bioavailability of the drug, and has good process feasibility and is suitable for industrial production. SUMMARY

[0007] Therefore, the purpose of the present application is to provide a preparation method of Dieda Qili preparation based on raw material particle size control, which can realize the simultaneous improvement of bioavailability and industrial production feasibility by accurately controlling the specific crushing particle size range of key medicinal ingredients without using complex processes.

[0008] To achieve the above-mentioned purpose, the scheme of the present application is as follows: The present application provides a preparation method of Dieda Qili preparation based on raw material particle size control, which is made of the following medicinal raw materials in mass fraction: artificial musk 0.8 parts, Sanqi 8 parts, dragon's blood 16 parts, safflower 48 parts, cinnabar 40 parts, wine Danggui 80 parts, vinegar myrrh 32 parts, borneol 1.6 parts, vinegar olibanum 32 parts and catechu 40 parts; the preparation method comprises the following steps: S1. Raw material pretreatment and crushing: crushing each medicinal raw material, wherein the crushing particle size of Sanqi is controlled between 80 mesh and 120 mesh; the crushing particle size of dragon's blood is controlled between 60 mesh and 100 mesh; S2. Total mixing and preparation: mixing the crushed medicinal raw materials to obtain a mixture, and preparing the mixture into the Dieda Qili preparation.

[0009] Alternatively, the crushing particle size of Sanqi is controlled between 80 mesh and 110 mesh, preferably between 80 mesh and 100 mesh, and more preferably 80 mesh.

[0010] Alternatively, the crushing particle size of dragon's blood is controlled between 70 mesh and 90 mesh, preferably between 80 mesh and 90 mesh, and more preferably 80 mesh.

[0011] Alternatively, the crushing particle size of catechu is controlled between 110 mesh and 130 mesh, preferably between 120 mesh and 130 mesh.

[0012] Alternatively, the crushing particle size of safflower is controlled between 110 mesh and 130 mesh, preferably between 120 mesh and 130 mesh.

[0013] Alternatively, the crushing particle size of wine Danggui is controlled between 110 mesh and 130 mesh, preferably between 120 mesh and 130 mesh.

[0014] Alternatively, the crushing particle size of dragon's blood is controlled between 70 mesh and 90 mesh, preferably between 70 mesh and 80 mesh.

[0015] Optionally, the artificial musk has a crushing particle size controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh.

[0016] Optionally, the acetic myrrh has a crushing particle size controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh.

[0017] Optionally, the borneol has a crushing particle size controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh.

[0018] Optionally, the cinnabar has a crushing particle size controlled between 120 mesh and 200 mesh, preferably between 140 mesh and 200 mesh.

[0019] Optionally, the acetic frankincense has a crushing particle size controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh.

[0020] Optionally, the cinnabar is crushed by water flying method.

[0021] Optionally, the preparation is selected from a powder or a tablet.

[0022] Optionally, when the preparation is a tablet, the Didi Qili preparation further comprises an excipient.

[0023] Optionally, the excipient comprises at least one of magnesium stearate, starch, dextrin, talc, microcrystalline cellulose, povidone, hydroxypropyl cellulose, microcrystalline silica, and sodium carboxymethyl starch.

[0024] The present application also provides use of the Didi Qili preparation prepared according to the method as described above in preparation of orthopedic trauma products.

[0025] As described above, the method for preparing the Didi Qili preparation based on raw material particle size and the use of the present application have the following beneficial effects: Firstly, the present application discovers through systematic research that for the key medicinal ingredients of Sanqi and Xuezhi in Dieda Qili preparation, the dissolution of the medicinal ingredients is not linearly increased with the decrease of particle size (increase of mesh number), but there is a specific optimal particle size window (Sanqi: 80-120 mesh; Xuezhi: 60-100 mesh), but the dissolution of the components under different particle sizes is not much different. In addition, the present application further discloses that in this specific compound Dieda Qili, simply combining the effective components of each single herb at the optimal particle size determined separately cannot necessarily obtain the best overall therapeutic effect of the compound. Specifically, although the single herb research suggests that the saponin components of Sanqi are higher at 80 mesh, the compound pharmacodynamics experiment surprisingly confirms that when the particle size of Sanqi is controlled at 100 mesh and the particle size of Xuezhi is controlled at 80 mesh, the synergistic therapeutic effect is significantly better than that of the combination of both at 80 mesh (i.e. the optimal dissolution point of each single herb). This discovery goes beyond simple particle size optimization and achieves the best balance between bioavailability and the feasibility of industrial production.

[0026] In summary, the particle size control technology provided by the present application, on the one hand, ensures the effective dissolution of key medicinal ingredients, thereby guaranteeing and improving the bioavailability and the final therapeutic effect of the preparation; on the other hand, the particle size range is appropriate, avoiding a series of process problems such as high energy consumption, poor material flowability, easy moisture absorption and hardening, and difficult tabletting caused by pursuing ultra-fine grinding, making the preparation process more stable and the cost lower, which has great advantages in industrial production. DETAILED DESCRIPTION

[0027] The embodiments of the present application are described below through specific examples, and those skilled in the art can easily understand other advantages and effects of the present application from the disclosure. The present application can also be implemented or applied through other different embodiments, and various modifications or changes can be made to the details in the specification without departing from the spirit of the present application.

[0028] An embodiment of the present application provides a preparation method of Dieda Qili preparation based on raw material particle size control, which is prepared from the following medicinal raw materials in parts by mass: artificial musk 0.8 parts, Sanqi 8 parts, Xuezhi 16 parts, Honghua 48 parts, Zhu Sha 40 parts, Jiudanggui 80 parts, Cudimahu 32 parts, Bingpian 1.6 parts, Cudilu 32 parts and Echa 40 parts; the preparation method of the Dieda Qili preparation comprises the following steps: S1. raw material pretreatment and crushing: crushing each medicinal raw material, wherein the crushing particle size of Panax notoginseng is controlled between 80 mesh and 120 mesh, preferably between 80 mesh and 110 mesh, further preferably between 80 mesh and 100 mesh, further more preferably 80 mesh; the crushing particle size of dragon's blood is controlled between 60 mesh and 100 mesh, preferably between 70 mesh and 90 mesh, further preferably between 80 mesh and 90 mesh, further more preferably 80 mesh; the crushing particle size of Fructus Evodiae is controlled between 110 mesh and 130 mesh, preferably between 120 mesh and 130 mesh; the crushing particle size of Carthamus tinctorius L. is controlled between 110 mesh and 130 mesh, preferably between 120 mesh and 130 mesh; the crushing particle size of Radix Angelicae Sinensis is controlled between 110 mesh and 130 mesh, preferably between 120 mesh and 130 mesh; the crushing particle size of dragon's blood is controlled between 70 mesh and 90 mesh, preferably between 70 mesh and 80 mesh; the crushing particle size of artificial musk is controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh; the crushing particle size of myrrh in vinegar is controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh; the crushing particle size of borneol is controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh; cinnabar is crushed by water flying method, and the crushing particle size of cinnabar is controlled between 120 mesh and 200 mesh, preferably between 140 mesh and 200 mesh; the crushing particle size of olibanum in vinegar is controlled between 60 mesh and 80 mesh, preferably between 70 mesh and 80 mesh; S2. total mixing and preparation: mixing the crushed medicinal raw materials to obtain a mixture, and preparing the mixture into a Qizhong Qili preparation; The preparation is selected from a powder or a tablet. When the preparation is a tablet, the Qizhong Qili preparation further comprises an excipient, and the excipient comprises at least one of magnesium stearate, starch, dextrin, talc, microcrystalline cellulose, povidone, hydroxypropyl cellulose, colloidal silicon dioxide and sodium carboxymethyl starch.

[0029] Another embodiment of the present application also provides use of the Qizhong Qili preparation prepared according to the method as described above in preparation of an orthopedic trauma product.

[0030] The present application will be described in detail below through specific examples. It should also be understood that the following examples are only used to specifically describe the present application, and cannot be understood as a limitation on the protection scope of the present application. Some non-essential improvements and adjustments made by those skilled in the art according to the above description of the present application all belong to the protection scope of the present application. The specific process parameters and the like described below are only one example in the appropriate range, i.e., those skilled in the art can make appropriate selection within the range through the description herein, and are not limited to the specific values of the examples below.

[0031] Example 1 The purpose of this example is to separately investigate the crushing particle size of Fructus Evodiae, Panax notoginseng, Carthamus tinctorius L., Radix Angelicae Sinensis and dragon's blood, which is as follows: It should be particularly pointed out that each sample is tested in triplicate, and the experimental results are expressed as the average value: (1) Investigation of the crushing particle size of catechu powder The same batch of catechu was divided into three parts, which were ground into catechu powder with a particle size of 80 mesh, catechu powder with a particle size of 100 mesh, and catechu powder with a particle size of 120 mesh, respectively. The contents of catechin and epicatechin in catechu powder with different particle sizes were detected according to the high performance liquid chromatography method of Chinese Pharmacopoeia (General 0512), and the total content of catechin and epicatechin was calculated. The results are shown in Table 1.

[0032] Table 1 Content of catechin and epicatechin in catechu powder with different particle sizes

[0033] As can be seen from Table 1, compared with 80 mesh catechu powder, the total content of catechin and epicatechin in 100 mesh catechu powder and 120 mesh catechu powder is increased, especially in 120 mesh catechu powder. The results show that the smaller the crushing particle size of the medicinal material, the higher the total content of catechin and epicatechin in catechu powder.

[0034] (2) Investigation of the crushing particle size of notoginseng powder The same batch of notoginseng was divided into three parts, which were ground into notoginseng powder with a particle size of 80 mesh, notoginseng powder with a particle size of 100 mesh, and notoginseng powder with a particle size of 120 mesh, respectively. The contents of ginsenoside Rg1, ginsenoside Rb1 and notoginsenoside R1 in notoginseng powder with different particle sizes were detected according to the high performance liquid chromatography method of Chinese Pharmacopoeia (General 0512), and the total content of ginsenoside Rg1, ginsenoside Rb1 and notoginsenoside R1 was calculated. The results are shown in Table 2.

[0035] Table 2 Content of ginsenoside Rg1, ginsenoside Rb1 and notoginsenoside R1 in notoginseng powder with different particle sizes

[0036] As can be seen from Table 2, compared with 120 mesh notoginseng powder, the total content of ginsenoside Rg1, ginsenoside Rb1 and notoginsenoside R1 in 80 mesh notoginseng powder and 100 mesh notoginseng powder is significantly increased, especially in 80 mesh notoginseng powder. The results show that for notoginseng, the smaller the crushing degree, the larger the particle size of the drug, and the dissolution degree of ginsenoside Rg1, ginsenoside Rb1 and notoginsenoside R1 in notoginseng powder is affected. However, it is generally believed by those skilled in the art that the smaller the crushing particle size of the drug raw material, the larger the specific surface area, the higher the dissolution rate, and the better the efficacy.

[0037] (3) Investigation of the crushing particle size of safflower powder The same batch of safflower was divided into three parts, and the three parts were ground into safflower powder with a particle size of 80 mesh, safflower powder with a particle size of 100 mesh and safflower powder with a particle size of 120 mesh respectively. The content of hydroxysafflor yellow A in safflower powder with different particle sizes was detected according to the high performance liquid chromatography method of Chinese Pharmacopoeia (general 0512), and three parallel tests were performed. The average value of the three parallel tests was taken as the detection result, and the results are shown in Table 3.

[0038] Table 3 Content of hydroxysafflor yellow A in safflower powder with different particle sizes

[0039] As can be seen from Table 3, compared with 80 mesh safflower powder, the content of hydroxysafflor yellow A in 100 mesh safflower powder and 120 mesh safflower powder is significantly increased, especially in 120 mesh safflower powder. The experimental results show that the smaller the particle size, the higher the content of hydroxysafflor yellow A in safflower powder.

[0040] (4) Investigation of the crushing particle size of wine angelica The same batch of wine angelica was divided into three parts, and the three parts were ground into angelica powder with a particle size of 80 mesh, angelica powder with a particle size of 100 mesh and angelica powder with a particle size of 120 mesh respectively. The content of ferulic acid in angelica powder with different particle sizes was detected according to the high performance liquid chromatography method of Chinese Pharmacopoeia (general 0512), and the results are shown in Table 4.

[0041] Table 4 Content of ferulic acid in angelica powder with different particle sizes

[0042] As can be seen from Table 4, compared with 80 mesh angelica powder, the content of ferulic acid in 100 mesh angelica powder and 120 mesh angelica powder is increased, especially in 120 mesh angelica powder. The results show that the smaller the particle size, the higher the content of ferulic acid in angelica powder.

[0043] (5) Investigation of the crushing particle size of blood The same batch of blood was divided into three parts, and the three parts were ground into blood powder with a particle size of 80 mesh, blood powder with a particle size of 100 mesh and blood powder with a particle size of 120 mesh respectively. The content of blood in blood powder with different particle sizes was detected according to the high performance liquid chromatography method of Chinese Pharmacopoeia (general 0512), and the results are shown in Table 5.

[0044] Table 5 Content of blood in blood powder with different particle sizes

[0045] As can be seen from Table 5, compared with the 120-mesh dragon's blood powder, the content of dracoquinone in the 80-mesh and 100-mesh dragon's blood powder is significantly increased, especially in the 80-mesh dragon's blood powder. The results show that for dragon's blood, the smaller the degree of crushing, the larger the particle size of the drug, and the dissolution degree of dracoquinone in the dragon's blood powder is affected. It is generally believed by those skilled in the art that the smaller the crushing particle size of the drug raw material, the larger the specific surface area, the higher the dissolution rate, and the better the drug efficacy.

[0046] The experimental results of this embodiment show that different drug raw materials have their own unique optimal dissolution particle size window, and the finer the particle size, the higher the dissolution rate of all components is not true. This is one of the most core findings of the present application, which is as follows: For catechu, safflower and wine danggui, the dissolution rate of the active ingredients (catechin and epicatechin, hydroxyl safflower yellow A, and ferulic acid) increases steadily as the particle size becomes finer (from 80 mesh to 120 mesh).

[0047] However, for the key medicinal herbs of panax notoginseng and dragon's blood, the dissolution rate of the core active ingredients (ginsenoside Rg1, Rb1, notoginsenoside R1, and dracoquinone) reaches a peak in the range of 80 mesh to 100 mesh, and when the particle size is further refined to 120 mesh, the content does not increase but decreases. This phenomenon is different from the generally accepted understanding that the finer the crushing, the better the dissolution. Therefore, whether the crushing particle size of different medicinal materials in the Didi Qili preparation will affect the bioavailability of the entire prescription needs further study.

[0048] Example 2 Based on the optimal particle size of each drug in Example 1, the purpose of this embodiment is to prepare a Didi Qili powder, which is as follows: The Didi Qili powder is made of the following drug raw materials in parts by mass: artificial musk 0.8 parts, panax notoginseng 8 parts, dragon's blood 16 parts, safflower 48 parts, cinnabar 40 parts, wine danggui 80 parts, vinegar opoponax 32 parts, borneol 1.6 parts, vinegar frankincense 32 parts, and catechu 40 parts. The preparation method of the Didi Qili powder is as follows: S1. Raw material pretreatment and crushing: The drug is ground according to the particle size in Table 6: Table 6 Crushing particle size table

[0049] S2. Total mixing and preparation: Mix 0.8 g of artificial musk powder, 8 g of panax notoginseng powder, 16 g of dragon's blood powder, 48 g of safflower powder, 40 g of cinnabar powder, 80 g of wine danggui powder, 32 g of vinegar opoponax powder, 1.6 g of borneol powder, 32 g of vinegar frankincense powder, and 40 g of catechu powder to obtain a mixture. The mixture is dried at a temperature of 50°C until the water content is 7%, to obtain the Didi Qili powder.

[0050] Example 3 Please refer to Example 1, the purpose of this example is: based on the optimal particle size in Example 1, the purpose of this example is to prepare a Qili pill, which is made of the following drug materials in mass fraction: artificial musk 0.8 parts, Sanqi 8 parts, dragon's blood 16 parts, safflower 48 parts, cinnabar 40 parts, wine Danggui 80 parts, vinegar myrrh 32 parts, borneol 1.6 parts, vinegar olibanum 32 parts and catechu 40 parts; The preparation method of the tablet provided in this example is as follows: S1. Pretreatment and crushing of raw materials: According to the particle size in Table 6, the drugs are ground: S2. Total mixing and preparation: Mix artificial musk powder 0.8 g, Sanqi powder 8 g, dragon's blood powder 16 g, safflower powder 48 g, cinnabar powder 40 g, wine Danggui powder 80 g, vinegar myrrh powder 32 g, borneol powder 1.6 g, vinegar olibanum powder 32 g, catechu powder 40 g and talc powder 9 g as an auxiliary material. Mix the above drugs and auxiliary materials to obtain a mixture, prepare granules, and spray dry the mixture under the conditions of inlet temperature 80℃ and material temperature 50℃ to a water content of 7%. Finally, add 4% of the total mass of magnesium stearate to the granules, and press the tablets to obtain Qili pill.

[0051] Test Example 1 The purpose of this test is: based on the previous research, Sanqi and dragon's blood are the core blood-activating and pain-relieving medicinal ingredients in this compound prescription. The dissolution of their active ingredients (ginsenosides Rg1 / Rb1 / sanqi saponin R1 and dragon's blood) does not increase linearly with the refinement of particle size, but peaks in the 80-100 mesh interval. This phenomenon is contrary to the conventional understanding, and both of them are large in dosage and strong in efficacy in the prescription, so their bioavailability is crucial to the overall efficacy.

[0052] Therefore, this test aims to observe the effect of different particle size combinations on the pharmacodynamics of Qili powder in animal models, and to investigate and confirm whether the crushing intensity of "Sanqi" and "dragon's blood" has a significant effect on the efficacy of the Qili preparation. The specific process is as follows: 1.1 Experimental animals Healthy adult SPF SD rats 120, half male and half female, weighing 180-220g, purchased from Chongqing Medical University. Using random number table method, divided into 2 groups: 10 in the sham operation group, 110 in the modeling group, during the experiment, fed in standard laboratory conditions (temperature 22±2℃, humidity 50±10%, 12 / 12h light-dark cycle) in separate cages, free access to standard feed and drinking water.

[0053] 1.2 Main reagents and instruments Nine kinds of Qidadi Qilil tablets were prepared respectively, specifically including: Tablet 1 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Sanqi was crushed and passed through a 100-mesh sieve); Tablet 2 (Qidadi Qilil tablets were prepared according to Example 3, except that the Sanqi was crushed and passed through an 80-mesh sieve, and the Xuejie was crushed and passed through an 80-mesh sieve); Tablet 3 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Sanqi was crushed and passed through a 120-mesh sieve); Tablet 4 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Xuejie was crushed and passed through a 100-mesh sieve); Tablet 5 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Xuejie was crushed and passed through a 120-mesh sieve); Tablet 6 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Sanqi was crushed and passed through a 100-mesh sieve and the Xuejie was crushed and passed through a 100-mesh sieve); Tablet 7 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Sanqi was crushed and passed through a 100-mesh sieve and the Xuejie was crushed and passed through a 120-mesh sieve); Tablet 8 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Sanqi was crushed and passed through a 120-mesh sieve and the Xuejie was crushed and passed through a 100-mesh sieve); Tablet 9 (Qidadi Qilil tablets were prepared in the same way as in Example 3, except that the Sanqi was crushed and passed through a 120-mesh sieve and the Xuejie was crushed and passed through a 120-mesh sieve); Each of the above tablets was ground and prepared into a suspension of the required concentration with a suitable solvent for easy administration; Free fall impact device (including guide pipe, weight, flat bottom impact head), small animal anesthetizing machine, electronic balance, vernier caliper, Von Frey fiber, enzyme label instrument, etc., sodium pentobarbital, paraformaldehyde, ELISA detection kit, etc.

[0054] 1.3 Model establishment The model group rats were treated as follows: The rat soft tissue contusion model was constructed by free fall impact method. After the rats were anesthetized by intraperitoneal injection of sodium pentobarbital (40-50 mg / kg), they were fixed in a prone position, the hair on the bilateral calf muscle area was shaved and disinfected. A 0.2 kg weight was dropped from a height of 20 cm, and the midpoint of the exposed gastrocnemius muscle was impacted, and the same operation was performed for 3 times, causing standardized acute soft tissue contusion. The sham operation group only received anesthesia and skin preparation, and did not receive impact modeling. A successful model should have the characteristics of local tissue indentation, rapid swelling, skin temperature rise, and subcutaneous ecchymosis. After the modeling was completed, the rats were individually fed under standard conditions, and their vital signs were closely monitored.

[0055] 1.4 Grouping and administration The 100 modeling successes were divided into 10 groups by random number table method, namely: group A, group B, group C, group D, group E, group F, group G, group J, group I and group H, 10 in each group, starting to give intragastric administration 3h after modeling success, once a day, for 7 consecutive days. Among them, group J is the model control group, the sham operation group and group J rats are given the same volume of pure water; the rest of the groups are given clinical equivalent dose of drug suspension.

[0056] Dosing: The clinical usage and dosage of Diedi Qili Tablets is oral, 1-3 tablets at a time, 3 times a day, and the daily crude drug amount is 0.8952g-2.6856g. Specifically, the dosing amount of this time is 0.16g of crude drug / kg, among which, group A is administered with suspension formed by tablet No.1, group B is administered with suspension formed by tablet No.2, group C is administered with suspension formed by tablet No.3, group D is administered with suspension formed by tablet No.4, group E is administered with suspension formed by tablet No.5, group F is administered with suspension formed by tablet No.6, group G is administered with suspension formed by tablet No.7, group H is administered with suspension formed by tablet No.8, and group I is administered with suspension formed by tablet No.9.

[0057] 1.5 Index observation 1.5.1 Gross observation score Before drug intervention (d0) and after drug intervention (d3, d7), the swelling and blood stasis of the contusion were observed, and the scores of the rats in each group were scored according to the scoring standard of Zhou Guolin et al., as shown in Table 7: Table 7 Gross observation score of rats

[0058] Note: If the classification is clear in the evaluation, the corresponding score is recorded, and if it is between, it can be evaluated as 1 or 3.

[0059] 1.5.2 Foot sole thickness The thickness of the right hind foot sole of the rats in each group was measured before drug intervention (d0) and after drug intervention (d3, d7) using an electronic vernier caliper; 1.5.3 Mechanical withdrawal threshold The changes of the mechanical withdrawal threshold of the right hind foot sole of the rats in each group before drug intervention (d0) and after drug intervention (d3, d7) were detected using Von Frey fiber; 1.5.4 Inflammatory factor and pain mediator level On the 7th day of administration (i.e. d7), the rats were sacrificed 1h after administration, and the injured limb muscle tissue was collected, and the levels of TNF-α, IL-1β, IL-10, 5-HT, β-EP and other factors in the injured tissue were detected by ELISA method.

[0060] 1.6 Statistical analysis SPSS 25.0 software was used for data processing. All measurement data were expressed as mean ± standard deviation (SD). One-way ANOVA was used for comparison among multiple groups. LSD method was used for pairwise comparison between groups if variances were equal, and Dunnett's T3 method was used if variances were not equal. P<0.05 was considered statistically significant.

[0061] 1.7 Experimental results and their analysis (1) The experimental results of gross observation and scoring are shown in Table 8: Table 8 Comparison of gross observation scores in each group (n=10) ±SD, n=10)

[0062] Note: Compared with the sham operation group # P<0.05; compared with group J * P<0.05, d7 represents 7 days after administration, d3 represents 3 days after administration, and so on.

[0063] After observation, obvious swelling and subcutaneous ecchymosis appeared at the injury site after modeling, and the unilateral hind limb was lameness and continuously aggravated, but there was no skin damage, obvious fracture dislocation characteristics. The sham operation group did not have obvious redness, ecchymosis and behavior disorders during the entire experimental process.

[0064] As shown in Table 8, before drug intervention (i.e. d0), the gross observation scores of all modeling groups were significantly higher than those of the sham operation group (P<0.05). Three days after administration (i.e. d3), compared with group J, the scores of each treatment group showed a downward trend, among which the scores of group A (Sanqi 100 meshes / Xueshuaiteng 80 meshes), group B (Sanqi 80 meshes / Xueshuaiteng 80 meshes), group D and group G improved most significantly (P<0.05). Seven days after administration (i.e. d7), the scores of most administration groups had returned to the 0-4 score interval, while group J still maintained at 4-6. Compared with group J, the scores of group A, group B, group D, group E, group G, group H and group I were significantly lower (P<0.05). Most importantly, the scores of group A (Sanqi 100 meshes / Xueshuaiteng 80 meshes) at all observation time points were the lowest among all treatment groups, showing the best recovery effect.

[0065] (2) The experimental results of foot thickness are shown in Table 9: Table 9 Comparison of foot thickness in each group (n=10) ±SD, n=10)

[0066] Note: Compared with the sham operation group # P<0.05; compared with group J​* P<0.05.

[0067] As shown in Table 9, before the administration intervention (i.e. d0), the toe thickness of all model rats was significantly increased, which was statistically different from the sham operation group (P<0.05), indicating that the modeling was successful. Over time, the swelling degree of the toes of each treatment group showed different degrees of reduction. Three days after administration (i.e. d3), compared with the J group, the toe thickness of the A group (Sanqi 100 meshes / Xuejie 80 meshes), the B group (Sanqi 80 meshes / Xuejie 80 meshes), the D group, the E group, the G group and the I group was significantly reduced (P<0.05). Seven days after administration (i.e. d7), the swelling of most of the treatment groups basically subsided, among which the toe thickness of the A group, the B group, the C group, the D group, the E group, the G group, the H group and the I group was significantly lower than that of the J group (P<0.05), and the recovery effect of the A group (Sanqi 100 meshes / Xuejie 80 meshes) was the most significant, and the d7 toe thickness was the smallest among all groups, which was always better than that of other groups.

[0068] (3) The results of the mechanical paw withdrawal threshold test are shown in Table 10: Table 10 Comparison of mechanical paw withdrawal threshold of each group ±SD, n=10

[0069] Note: compared with the sham operation group # P<0.05; compared with the J group * P<0.05.

[0070] As shown in Table 10, before the administration intervention (i.e. d0), the mechanical paw withdrawal threshold of all model rats was significantly lower than that of the sham operation group (P<0.05), indicating that the mechanical pain threshold was decreased and the modeling was successful. During the treatment, the recovery degree of the pain threshold of each administration group showed differences. Three days after administration (i.e. d3), the mechanical paw withdrawal threshold of the H group and the F group showed a downward trend, but there was no significant difference, and the mechanical paw withdrawal threshold of the remaining administration groups (including the A group and the B group) was significantly higher than that of the J group (P<0.05). Seven days after administration (i.e. d7), the mechanical paw withdrawal threshold of each administration group was significantly higher than that of the J group (P<0.05). Among them, the mechanical paw withdrawal threshold of the A group (Sanqi 100 meshes / Xuejie 80 meshes) at d3 and d7 was significantly higher than that of all other treatment groups, showing the strongest analgesic effect.

[0071] (4) The results of the inflammatory factor level test are shown in Table 11: Table 11 Comparison of inflammatory factor levels in tissues of rats in each group ±SD, n=10

[0072] Note: compared with the sham operation group #P<0.05; compared with group J * P<0.05.

[0073] From Table 11, the ELISA detection results show that the levels of TNF-a and IL-1β in the local tissues of the injury site significantly increase (P<0.05) and the level of IL-10 significantly decreases (P<0.05) after modeling. After the drug intervention, the levels of the pro-inflammatory factors in each drug group are reduced to different degrees, and the anti-inflammatory factor is significantly increased. The group A (100 mesh of Panax notoginseng / 80 mesh of Sanguis Draconis) has the most significant effect in reducing the pro-inflammatory factors TNF-a and IL-1β and increasing the anti-inflammatory factor IL-10, and the levels of TNF-a and IL-1β are the lowest and the level of IL-10 is the highest in all treatment groups. Except for the group C, the levels of TNF-a in the rest of the drug groups are significantly lower than those in the group J (P<0.05); the levels of IL-1β in each drug group are significantly lower than those in the group J (P<0.05) and the level of IL-10 is significantly increased (P<0.05).

[0074] (5) The experimental results of the levels of pain mediators are shown in Table 12: Table 12 Comparison of the levels of pain mediators in the tissues of rats in each group ±SD, n=10

[0075] Note: compared with the sham operation group # P<0.05; compared with group J * P<0.05.

[0076] From Table 12, the ELISA detection results show that the levels of 5-HT and PGE2 in the local tissues of the injury site significantly increase (P<0.05) and the level of β-EP significantly decreases (P<0.05) after modeling. After the drug intervention, the levels of the pain mediators in each drug group are reduced to different degrees, and the analgesic substance level is significantly increased. The group A (100 mesh of Panax notoginseng / 80 mesh of Sanguis Draconis) has the best performance in regulating the pain mediators, and the level of β-EP is significantly higher than that in the group J and most of the treatment groups, while the level of PGE2 is closest to the normal level of the sham operation group. The levels of 5-HT and PGE2 in each drug group are significantly lower than those in the group J (P<0.05); at the same time, the levels of β-EP in the groups H, E, and F have an increasing trend, but there is no statistical significance, and the levels of β-EP in the groups A, B, C, D, G, and I are significantly higher than those in the group J (P<0.05).

[0077] The above experimental results indicate that the tablets prepared from Group A (100 mesh Panax notoginseng / 80 mesh Dragon's Blood) exhibit comprehensive and significant advantages in treating acute soft tissue contusions. Specifically, compared with the model control group (Group J) and other tablets with different particle size combinations, Group A most rapidly and effectively improved limb swelling (minimum plantar thickness), restored functional activity (lowest gross score), and increased the mechanical pain threshold (strongest analgesic effect). It also optimally regulated the local inflammatory response (significantly reduced TNF-α and IL-1β, increased IL-10) and pain mediator levels (significantly regulated 5-HT, PGE2, and β-EP to near normal levels). These data clearly reveal the dose-response relationship between efficacy and the particle size distribution of Panax notoginseng and Dragon's Blood. Most importantly, this invention has found that although studies on the dissolution of single herbal components suggest that the saponin components of Panax notoginseng dissolve more at 80 mesh (see Example 1), pharmacodynamic experiments of compound preparations show that when the particle size of Panax notoginseng is controlled at 100 mesh and that of dragon's blood is controlled at 80 mesh (Group A), the overall therapeutic effect is significantly better than when both Panax notoginseng and dragon's blood are pulverized at 80 mesh (Group B) or any other combination of particle sizes.

[0078] Experimental Example 2 This test aims to observe whether the key quality attributes (friability, weight variation, and disintegration time) of the Die Da Qi Li tablets prepared using the preferred particle size parameters of this invention (based on the 100 mesh of Panax notoginseng and 80 mesh of Dragon's Blood determined in Test Example 1) meet the requirements of the Chinese Pharmacopoeia, as detailed below: 2.1 Test Instruments 30B Small and Medium-Sized Crusher (Fujian Zhiluo Technology Co., Ltd.); ZXRD-A7140 Constant Temperature Drying Oven (Shanghai Zhicheng). YXQ.MG—Ⅱ-S-00 Sterilization Cabinet (Sichuan Shehong General Medical); GTC100 granulator (Hebei Ao'an); ST-B200 Micro Sample Grinding Machine (Fujian Zhiluo Technology Co., Ltd.); ZP8ES tablet press (Shanghai Xinyuan Pharmaceutical Machinery Co., Ltd.); ZB-1E Intelligent Disintegration Analyzer (Tianjin Tianda Tianfa Co., Ltd.); YD-35 tablet hardness tester (Tianjin Tianda Tianfa Co., Ltd.).

[0079] 2.2 Formulation and Preparation Process This experimental example demonstrates the preparation of Die Da Qi Li tablets according to the following specific steps:

Prescription

[0080] Preparation method S1. Raw material pretreatment and pulverization: The drug was ground according to the particle size in Table 13: Table 13 Particle size of each drug in the prescription of Test Example 2

[0081] S2. Total mixing and preparation: mixed artificial musk powder 0.8 g, Sanqi powder 8 g, dragon's blood powder 16 g, safflower powder 48 g, cinnabar powder 40 g, wine Danggui powder 80 g, vinegar myrrh powder 32 g, borneol powder 1.6 g, vinegar olibanum powder 32 g, and catechu powder 40 g to obtain a mixture, and the mixture was dried at a temperature of 50°C until the water content was 7% to obtain the Didi Qili powder.

[0082] S2. Total mixing and preparation: mixed artificial musk powder 0.8 g, Sanqi powder 8 g (passed through a 100-mesh sieve), dragon's blood powder 16 g, safflower powder 48 g, cinnabar powder 40 g (passed through a 200-mesh sieve), wine Danggui powder 80 g, vinegar myrrh powder 32 g, borneol powder 1.6 g, vinegar olibanum powder 32 g, catechu powder 40 g, and talcum powder 9 g to obtain a mixture, and the mixture was granulated by dry granulation process, the granules were sieved with a 16-mesh sieve, the mixture was spray dried at an inlet temperature of 80°C and a material temperature of 50°C until the water content was 7%, and then the tablets were compressed by a conventional method (5% of the total weight of magnesium stearate was added to the qualified granules as a lubricant, mixed uniformly, and compressed into tablets by a ZP8ES tablet press, with a tablet weight of 0.30 g and a compression pressure of 15 KN) to obtain the Didi Qili tablets.

[0083] 2.3 Tablet quality inspection results The Didi Qili tablets prepared according to the above method were subjected to the following key quality inspections: (1) Friability inspection: an appropriate amount of tablets were placed in the cylinder of the tablet friability inspection instrument and rotated for 100 times, and no cracking, cracking, or crushing phenomenon occurred. Calculation showed that the friability was 0.65%, which was lower than the limit of 1.0% specified in the Chinese Pharmacopoeia, meeting the requirements.

[0084] (2) Weight difference inspection: 20 tablets were randomly selected for inspection, and the tablet weight difference range was between -2.2% and +2.6%, and no tablets exceeding the tablet weight difference limit (±5%) were found, meeting the requirements of the Chinese Pharmacopoeia for the weight difference of 0.30 g tablets.

[0085] (3) Disintegration time inspection: all tablets were completely disintegrated within 15 min according to the disintegration time inspection method in the Chinese Pharmacopoeia 0921, meeting the requirements.

[0086] The test example results show that the Didi Qili tablets prepared by using the preferred particle size combination of 100 mesh for Panax notoginseng and 80 mesh for dragon's blood and the corresponding preparation process meet the Chinese Pharmacopoeia standards in key quality indicators such as friability, weight difference and disintegration time, the process is stable and feasible, the preparation quality is excellent, and the requirements of industrial production can be met.

[0087] The above examples only illustrate the principles and effects of the present application, and are not intended to limit the present application. Any person skilled in the art can modify or change the above examples without departing from the spirit and scope of the present application. Therefore, all equivalent modifications or changes made by those skilled in the art without departing from the spirit and technical idea disclosed by the present application should be covered by the claims of the present application.

Claims

1. A preparation method of Didiqi Li preparation based on raw material particle size control, the Didiqi Li preparation is prepared from the following medicinal raw materials in mass fraction: artificial musk 0.8 parts, Sanqi 8 parts, dragon's blood 16 parts, safflower 48 parts, cinnabar 40 parts, wine Danggui 80 parts, vinegar Myrrh 32 parts, borneol 1.6 parts, vinegar Olibanum 32 parts and catechu 40 parts; characterized in that, The preparation method comprises the following steps: S1. Raw material pretreatment and crushing: crushing each medicinal raw material, wherein the crushing particle size of the Panax notoginseng is controlled between 80 meshes and 120 meshes; the crushing particle size of the dragon's blood is controlled between 60 meshes and 100 meshes; S2. Total mixing and preparation: mixing each crushed medicinal raw material to obtain a mixture, and preparing the mixture into the Didi Qili preparation.

2. The production method according to claim 1, wherein The crushing particle size of the Panax notoginseng is controlled between 80 meshes and 110 meshes; And / or, the crushing particle size of the dragon's blood is controlled between 70 meshes and 90 meshes.

3. The production method according to claim 1, wherein The crushing particle size of the Radix Paeoniae Alba is controlled between 110 meshes and 130 meshes; And / or, the crushing particle size of the Carthamus tinctorius L. is controlled between 110 meshes and 130 meshes.

4. The production method according to claim 1, wherein The crushing particle size of the Radix Angelicae Sinensis is controlled between 110 meshes and 130 meshes; And / or, the crushing particle size of the dragon's blood is controlled between 70 meshes and 90 meshes.

5. The production method according to claim 1, wherein The crushing particle size of the artificial musk is controlled between 60 meshes and 80 meshes; And / or, the crushing particle size of the myrrh in vinegar is controlled between 60 meshes and 80 meshes.

6. The production method according to claim 1, wherein The crushing particle size of the ice flakes is controlled between 60 meshes and 80 meshes.

7. The production method according to claim 1, wherein The crushing particle size of the cinnabar is controlled between 120 meshes and 200 meshes; And / or, the crushing particle size of the olibanum in vinegar is controlled between 60 meshes and 80 meshes; And / or, the cinnabar is crushed by water flying method.

8. The production method according to claim 1, wherein The preparation is selected from a powder or a tablet.

9. The production method according to claim 8, wherein When the preparation is a tablet, the Didi Qili preparation further comprises an excipient.

10. Use of the Didi Qili preparation prepared by the method according to any one of claims 1-9 in the preparation of an orthopedic trauma product.

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