External traditional Chinese medicine composition for promoting blood circulation to arrest pain as well as preparation method and application of external traditional Chinese medicine composition

Abstract

The invention relates to the field of traditional Chinese medicine compositions, and discloses an external traditional Chinese medicine composition for promoting blood circulation to arrest pain as well as a preparation method and application of the external traditional Chinese medicine composition. The fixed component is prepared from the following components in parts by weight: 20 to 40 parts of raw rhubarb and 20 to 40 parts of fructus gardeniae praeparatus; 8-16 parts of ground beetle, 10-20 parts of vinegar frankincense and 10-20 parts of vinegar myrrh; 15-25 parts of ligusticum wallichii and 6-14 parts of angelica sinensis. The traditional Chinese medicine composition for external use makes up the blank of heat syndrome treatment of the existing blood-activating and pain-relieving medicine by precisely targeting red, swollen and heat pain in the early stage of acute injury, and realizes heat-clearing and blood-stasis-removing integrated treatment through a synergistic effect; component permeation is accelerated through a specific ratio, and the problem of slow effect is solved; multi-effect components are further integrated, and medication is dynamically adjusted according to an acute phase and a recovery phase; clinical tests show that the 24-hour detumescence rate and the total effective rate are increased, the safety is good, the traditional Chinese medicine composition can be widely applied to acute soft tissue injury, chronic strain and other soft tissue injury diseases, and the curative effect is improved and the safety is guaranteed.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of traditional Chinese medicine compositions, in particular to a blood-activating and pain-relieving external traditional Chinese medicine composition and a preparation method and application thereof. BACKGROUND

[0002] Soft tissue injury is one of the most common diseases in clinical orthopedics, including acute closed soft tissue injury and chronic soft tissue strain, and its incidence accounts for more than 60% of orthopedic clinic patients. The disease is mainly manifested as local pain, swelling, blood stasis, and dysfunction, which seriously affects the quality of life and work ability of patients. Traditional Chinese medicine believes that the pathogenesis of soft tissue injury is qi stagnation and blood stasis, and the treatment principle is to activate blood and resolve stasis. External traditional Chinese medicine preparations have become an important means for treating soft tissue injury due to their advantages of directly reaching the disease site, fast effect, convenient use, and small side effects. With the increasing incidence of soft tissue injury, it is of great clinical significance to develop external traditional Chinese medicine preparations with definite efficacy and high safety.

[0003] At present, the existing product formula focuses on activating blood and resolving stasis, ignoring the inflammatory exudation and heat-toxin accumulation pathogenesis in the early stage (within 24-72 hours) of acute injury. Simple blood-activating is easy to aggravate local hyperemia and edema, and it is common for patients to have swelling after using blood-activating drugs. This is because of the lack of compatibility of heat-clearing and detumescence drugs. The commonly used external traditional Chinese medicine preparations for blood-activating and pain-relieving mainly include safflower oil, bone-setting water, dog skin plaster, and Yunnan white medicine aerosol. These products are mainly composed of simple blood-activating and resolving stasis traditional Chinese medicines, such as safflower oil mainly composed of safflower and cassia twig, bone-setting water mainly composed of earthworm, safflower, and angelica, and dog skin plaster mainly composed of frankincense, myrrh, and musk. The formula idea mainly focuses on general blood-activating and pain-relieving, and long-term efficacy of relaxing sinews and activating collaterals, ignoring the pathological characteristics of redness, swelling, heat, and pain, and inflammatory exudation and heat-toxin accumulation in the acute early stage of soft tissue injury. On this basis, the pathological process of soft tissue injury is dynamic, and the acute stage is mainly characterized by inflammatory reaction and microcirculation disorder, which requires both heat-clearing and detumescence and blood-activating. The recovery stage is mainly characterized by tissue repair and blood stasis absorption, which requires strengthening of blood-activating and resolving stasis. The existing products use fixed formula and unified method, which cannot adjust the treatment strategy according to the disease course, leading to problems of poor detumescence in the acute stage and slow repair in the recovery stage. SUMMARY

[0004] In order to solve the problems of lack of targeted treatment, slow effect and single efficacy composition which is difficult to adapt to the pathological characteristics of different stages of injury in the early stage of acute injury in the prior art, the present application provides a traditional Chinese medicine external composition with the functions of promoting blood circulation and relieving pain and clearing heat and swelling, and a preparation method and application thereof. The traditional Chinese medicine external composition targets the redness, swelling, heat and pain in the early stage of acute injury, fills the heat syndrome treatment blank of the existing blood-activating and pain-relieving medicaments, and realizes integrated treatment of heat clearing and stasis resolving through synergistic effect. Moreover, the composition penetration is accelerated through specific proportioning, so as to solve the problem of slow effect. Further, multi- efficacy components are integrated and the medication is dynamically adjusted according to the acute stage and recovery stage. Through clinical trials, the present application improves the 24-hour swelling reduction rate and total effective rate, and has good safety and can be widely applied to soft tissue injury diseases such as acute soft tissue injury and chronic strain.

[0005] In one aspect, the present application provides a traditional Chinese medicine external composition for promoting blood circulation and relieving pain, which comprises fixed components and replaceable components. In terms of weight parts, The fixed components comprise 20-40 parts of raw rhubarb and 20-40 parts of fried gardenia; 8-16 parts of eupolyphaga, 10-20 parts of vinegar olibanum and 10-20 parts of vinegar myrrh; 15-25 parts of chuanxiong and 6-14 parts of angelica. The replaceable components are one of the following three combinations: Combination one: 6-14 parts of safflower, 8-16 parts of sparganium stoloniferum, 8-16 parts of curcuma zedoary and 8-16 parts of vinegar corydalis; Combination two: 8-12 parts of turmeric, 10-16 parts of salvia miltiorrhiza, 6-10 parts of lindera and 6-10 parts of costus; Combination three: 10-20 parts of litchi kernel, 6-10 parts of curcuma zedoary, 1.5-3 parts of leech and 10-15 parts of cassia twig.

[0006] Further, the weight ratio of raw rhubarb to fried gardenia is 1:(0.8-1.2). Further, the weight ratio of raw rhubarb to fried gardenia is 1:1.

[0007] Further, the weight ratio of vinegar olibanum, vinegar myrrh to eupolyphaga is (1-1.5):(1-1.5):1. Further, the weight ratio of vinegar olibanum, vinegar myrrh to eupolyphaga is 1.25:1.25:1.

[0008] Further, the weight ratio of chuanxiong to angelica is (1.5-2.5):1. Further, the weight ratio of chuanxiong to angelica is 2:1.

[0009] Further, the external use traditional Chinese medicine composition further comprises one or more auxiliary medicinal materials: 10-20 parts by weight of Radix Sargentodoxae, 6-14 parts by weight of Radix Angelicae Pubescentis, 6-14 parts by weight of Radix Anemarrhenae, and 2-6 parts by weight of a transdermal enhancer selected from one or more of camphor, borneol, and musk. Further, the transdermal enhancer is camphor, and the camphor accounts for 1.5-2.5% of the total weight of the external use traditional Chinese medicine composition.

[0010] Further, the raw material parts by weight ratio is as follows: The heat-clearing and swelling-eliminating group comprises 25-35 parts of raw rhubarb, and 25-35 parts of fried gardenia fruit; The blood-activating and stasis-dissolving group comprises 10-14 parts of Eupolyphaga, 12-18 parts of vinegar myrrh, 12-18 parts of vinegar myrrh, 8-12 parts of safflower, 10-14 parts of rhizoma sparganii, 10-14 parts of curcuma zedoary, and 10-14 parts of vinegar Corydalis. The qi-regulating and pain-stopping group comprises 18-22 parts of Szechuan lovage rhizome, 8-12 parts of angelica, 12-18 parts of Radix Sargentodoxae, 8-12 parts of Radix Angelicae Pubescentis, and 8-12 parts of Radix Anemarrhenae. The transdermal enhancer comprises 3-5 parts of camphor.

[0011] Further, the total weight ratio of the heat-clearing and swelling-eliminating group to the blood-activating and stasis-dissolving group is 1: (1.5-1.8).

[0012] The second aspect of the present application provides a method for preparing a medicinal liquor from the external use traditional Chinese medicine composition of the present application, comprising the following steps: (1) weigh each raw material according to the parts by weight, and grind the raw materials except camphor to 10-20 mesh; (2) mix the heat-clearing and swelling-eliminating group and the qi-regulating and pain-stopping group, add 45-60% volume fraction of aqueous ethanol solution, and the solid-liquid ratio is 1: (8-12), and then soak at 20-30℃ for 10-20 days; mix the blood-activating and stasis-dissolving group, and separately soak under the same conditions; (3) combine the two groups of extraction solutions, filter through 180-220 mesh filter cloth, and collect the filtrate; (4) dissolve camphor in 95% volume fraction of aqueous ethanol solution with 20-30 times the weight of camphor, prepare a camphor alcohol solution, and add it at a rate of not more than 3% of the total volume of the filtrate per minute, and stir at a speed of 100-150 rpm at the same time; (5) stand for 12-48 hours, and filter.

[0013] Further, in step (2), the ethanol concentration is 52%.

[0014] Further, in step (2), the solid-liquid ratio is 1:10.

[0015] Further, in step (2), the soaking temperature is 25±2℃.

[0016] Further, in step (2), the leaching time is 14 days. Further, in step (2), the first 7 days are shaken 2 times a day for 5 minutes each, and the last 7 days are leaching with static.

[0017] Further, in step (3), the obtained residue is filtered and washed once with ethanol equivalent to 20-25% of the original volume, and the washing liquid is added to the filtrate.

[0018] The third aspect of the present application provides a method for preparing a traditional Chinese medicine external composition into a medicinal powder, characterized in that each kind of traditional Chinese medicine raw material is ground into 80-120 mesh fine powder, and mixed uniformly according to the formula proportion to prepare an external medicinal powder.

[0019] Further, the camphor is ground to more than 150 mesh, added and mixed uniformly.

[0020] The fourth aspect of the present application provides a use of a traditional Chinese medicine external composition as described in the present application in the preparation of an external medicine for treating soft tissue injury, which is used in the acute stage to mainly clear heat and reduce swelling, 3-4 times a day; and is used in the recovery stage to mainly promote blood circulation and remove blood stasis, 2-3 times a day.

[0021] Further, the soft tissue injury includes acute closed soft tissue injury, chronic soft tissue strain, muscle sprain and contusion, ligament sprain, contusion and swelling in early stage of fracture.

[0022] Further, the use of the medicine is local application, application or spraying.

[0023] The existing blood-activating and pain-relieving external preparation has the following key technical problems in clinical application: first, it lacks targeted treatment for redness, swelling and pain in the early stage of acute injury, second, the low transdermal absorption efficiency of effective components leads to slow effect, and third, the single efficacy formula is difficult to adapt to the pathological characteristics of different stages of injury. Compared with the prior art, the present application has at least the following beneficial effects: 1. In the present application, each traditional Chinese medicine is scientifically matched, solving the problems of weak and slow effect of single traditional Chinese medicine for activating blood and relieving pain, and the triple effects of “activating blood, relieving pain and transdermal” are achieved by the synergistic effect of each component, and there is no skin irritation and other side effects.

[0024] 2. The present application limits the weight ratio of the core components to a specific range, which is verified by experiments to be the optimal efficiency ratio, achieving the maximum efficacy.

[0025] 3. This invention employs a group extraction process, where heat-clearing herbs and blood-activating herbs are extracted separately and then combined. Heat-clearing herbs have a cooling effect, reducing the damage to soft tissues caused by inflammatory reactions, while blood-activating herbs can dilate microvessels and improve circulation. The combination of these two extracts helps improve local blood circulation, providing better blood supply to tissues, promoting tissue repair and regeneration, and solving the problems of low utilization rate and poor transdermal absorption of active ingredients in traditional external Chinese medicine. Furthermore, the process is simple, controllable, and suitable for industrial production.

[0026] 4. This invention precisely targets the early stages of acute injury with redness, swelling, heat, and pain by constructing a heat-clearing and swelling-reducing group, filling a gap in the treatment of heat syndromes; it uses camphor as a transdermal penetration enhancer to accelerate ingredient penetration and solve the problem of slow onset of action; it integrates multiple functional components and dynamically adjusts them according to the acute and recovery phases, overcoming the limitations of single-component formulations. Based on this, it enhances synergistic effects by optimizing the formulation ratio, achieving integrated treatment of heat clearing and blood stasis removal, thus improving efficacy and ensuring safety. Attached Figure Description

[0027] Figure 1 A flowchart illustrating the preparation method of the traditional Chinese medicine external application composition for promoting blood circulation and relieving pain provided in this embodiment of the invention; Figure 2 A line graph illustrating the anti-inflammatory effect evaluation provided in this embodiment of the invention; Figure 3 A line graph illustrating the comprehensive efficacy evaluation provided in this embodiment of the invention; Figure 4 A line graph showing the trend of time-based cumulative permeation of emodin, provided for embodiments of the present invention; Figure 5 A line graph illustrating the trend of acute-phase medication frequency versus swelling reduction rate, provided in an embodiment of the present invention; Figure 6 A line graph illustrating the trend of recovery effect changes in the frequency of medication during the recovery period, as provided in an embodiment of the present invention. Detailed Implementation

[0028] The endpoints and any values ​​of the ranges disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0029] In the present application, Shengdahuang is the dried root and rhizome of Rheum palmatum L., Rheum tanguticum Maxim. ex Balf. or Rheum officinale Baill.; Chaozhizi is the dried ripe fruit of Gardenia jasminoides Ellis, which is prepared by frying to brown; Dibugchong is the dried female body of Eupolyphaga sinensis Walker or Steleophaga plancyi (Boleny); Daxueteng is the dried stem of Sargentodoxa cuneata (Oliv.) Rehd. et Wils.

[0030] In the present application, the vinegar-lacquered frankincense, vinegar-lacquered myrrh and vinegar-lacquered Corydalis are prepared according to the vinegar-lacquered method in the Pharmacopoeia of the People's Republic of China; wherein, the vinegar-lacquering process is as follows: taking the cleaned medicinal materials, adding 15-20% rice vinegar by weight of the medicinal materials, mixing well, and moistening until the vinegar is absorbed, then placing in a frying container and frying on a low fire until the surface is shiny, and then taking out and cooling.

[0031] According to an embodiment of the present application, the use method of the medicinal powder comprises: taking 5-15g of the medicinal powder, and selecting a blending medium according to different situations: for those with obvious redness, heat and pain in acute stage: blending into paste with honey; for those with blood stasis in chronic stage: blending into paste with yellow rice wine or white wine; for those with sensitive skin: blending into ointment with vaseline or tea oil.

[0032] According to an embodiment of the present application, the soft tissue injury includes: acute closed soft tissue injury, chronic soft tissue strain, muscle sprain and contusion, ligament sprain, contusion and swelling in early stage of fracture.

[0033] According to an embodiment of the present application, the use mode of the medicine is topical application, pasting or spraying.

[0034] In the present application, in the preparation process, the traditional external use medicinal liquor usually adopts single ethanol extraction method, and all medicinal materials are mixed and extracted at one time. In the aspect of transdermal absorption promotion, the existing products mainly rely on the solvent effect and volatility of ethanol, or add traditional transdermal enhancers such as borneol and menthol. In the aspect of clinical application, the existing products usually do not distinguish acute stage and recovery stage, and adopt a unified medication scheme. However, the present application targets the early stage of acute injury by forming a heat-clearing and swelling-removing group, fills the blank of heat syndrome treatment; accelerates the penetration of components by using camphor as a transdermal enhancer, solves the problem of slow effect; breaks through the limitation of single group adaptation by integrating components with multiple effects and dynamically adjusting according to acute stage and recovery stage. On this basis, the synergistic effect is strengthened by optimizing the group ratio, realizing the integrated treatment of heat-clearing and blood-quickening, and having the effects of improving efficacy and ensuring safety.

[0035] The technical solutions of the present application will be described clearly and completely below with reference to the drawings. Obviously, the described embodiments are only some of the embodiments of the present application, but not all of them. Based on the embodiments of the present application, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of the present application.

[0036] The raw materials used in the following examples, if not specifically limited, are disclosed in the prior art, for example, can be directly purchased or prepared according to the preparation method disclosed in the prior art.

[0037] In the following examples, the traditional Chinese medicinal materials used meet the standards of the People's Republic of China Pharmacopoeia (2020 edition), including the following raw materials: Raw rhubarb (dried roots and rhizomes of Rheum palmatum L., purchased from Beijing Tong Ren Tang); Fried gardenia fruit (dried mature fruits of Gardenia jasminoides Ellis, fried to brown); Eupolyphaga (dried female bodies of Eupolyphaga sinensis Walker); Vinegar lactose (lactose resin prepared according to the vinegar frying method in the Pharmacopoeia); Vinegar myrrh (myrrh resin prepared according to the vinegar frying method in the Pharmacopoeia); Safflower (dried flowers of Carthamus tinctorius L.); Angelica (dried roots of Angelica sinensis (Oliv.) Diels); Chuanxiong rhizome (dried rhizomes of Ligusticum chuanxiong Hort.); Sargentodoxa cuneata (dried stems of Sargentodoxa cuneata (Oliv.) Rehd. et Wils. of the Lardizabalaceae family); Curcuma phaeocaulis (dried rhizomes of Curcuma phaeocaulis Val., Curcuma kwangsiensis S.G. Lee et C.F. Liang or Curcuma wenyujin Y.H. Chen et C.Ling of the Zingiberaceae family); Baishizhi (dried roots of Angelica dahurica (Fisch. ex Hoffm.) Benth. et Hook. f. or Angelica dahurica (Fisch. ex Hoffm.) Benth. et Hook. f. var. formosana (Boiss.) Shan et Yuan); Rhizoma Sparganii (dried rhizome of Sparganium stoloniferum Buch.-Ham.); Rhizoma Corydalis (prepared by vinegar-fried method according to the Pharmacopoeia); Radix Clematidis (dried root and rhizome of Clematis chinensis Osbeck, Clematis hexapetala Pall. or Clematis manshurica Rupr.); Camphor (pharmaceutical grade camphor, content ≥ 96%).

[0038] The specific process of vinegar-fried technology is as follows: take 100 g of Olibanum, Myrrh and Corydalis respectively, add 18 mL of rice vinegar (equivalent to 18% of the weight of medicinal materials), mix well, soak for 2 hours until the vinegar is absorbed, put into a frying pan, and fry on low heat (120-150°C) for 15-20 minutes until the surface is shiny, take out and cool.

[0039] Example 1 The formula is as follows: Heat-clearing and Detumescence Group: raw Dahuang 30 g, Jiaozishizao 30 g; Blood-activating and Stasis-removing Group: Tuobiechong 12 g, Olibanum 15 g, Myrrh 15 g, Honghua 10 g, Sanling 12 g, E'zhu 12 g, Corydalis 12 g; Qi-promoting and Pain-stopping Group: Chuanqiong 20 g, Danggui 10 g, Dahuangshu 15 g, Radix Clematidis 10 g, Baizhi 10 g; Transdermal Promoter: Camphor 4 g.

[0040] Alternative preparation process of the prescription: Group extraction method is adopted, and the steps include: Step S1, medicinal material pretreatment; grind all raw materials except camphor into 15-mesh coarse powder; Step S2, group extraction; the first group (heat-clearing and detumescence group + Qi-promoting and pain-stopping group) with a total weight of 125 g is added with 52% ethanol 1250 mL (solid-liquid ratio 1:10) in a sealed container, and constant temperature extraction is carried out at 25±2°C; the second group (blood-activating and stasis-removing group) with a total weight of 88 g is added with 52% ethanol 880 mL, and the same condition is used for extraction; on this basis, shake 2 times a day for 5 minutes each time in the first 7 days; and stand still in the last 7 days; Step S3, combine and filter; after 14 days, filter with 200-mesh filter cloth respectively; wash the residues with 250 mL of 52% ethanol once; and combine the washing liquid with the filtrate, about 2350 mL (there may be losses such as adsorption, filtration residue and volatilization of the residues) is obtained; Step S4, camphor treatment; dissolve camphor 4g in 100mL 95% ethanol, dissolve thoroughly; further, slowly add the filtrate at a speed of 50mL / min, stirring at a speed of 120rpm; Step S5, product preparation; continue stirring for 30 minutes, stand for 24 hours; filter again, get about 2400mL of clear medicinal liquor; divide into 100mL brown glass bottles, seal and store.

[0041] The quality test results of the prescription are shown in Table 1; Table 1 Example 2 Compared with Example 1, the dosages of other medicinal materials are fixed, and the ratio of raw rhubarb and fried gardenia is adjusted (one decimal place is retained), as shown in Table 2; Table 2 In this embodiment, further, an anti-inflammatory effect evaluation is performed, which can use the SD rat foot swelling model, cross foot inflammation, and measure the swelling inhibition rate after administration.

[0042] Anti-inflammatory and detumescence experiment (SD rat cross foot inflammation model): Grouping: 30 SD rats (male, 200-250g) were purchased from Jiangsu Jizhu Pharmaceutical Kang Biotechnology Co., Ltd. (License No. SCXK (Su) 2023-0009), and the feeding environment was temperature 22±2℃, humidity 55±5%, 12h light / dark cycle, and adaptive feeding for 3 days before experiment; randomly divided into blank group, control group and test group, 10 rats in each group.

[0043] Instruments: hot plate instrument (model: SA702), electronic vernier caliper (accuracy 0.01mm), rat fixing frame, cross foot inflammation device (self-made: containing anti-slip tread plate, adjustable limiting baffle, pressure sensor, tread plate area 10cm×10cm, can set the stepping pressure) Before the experiment, the right hind foot of each rat was measured with an electronic vernier caliper (take 1cm below the ankle joint, measure continuously for 3 times, and take the average value as the basic circumference L0); Cross foot inflammation operation (simulating clinical bruise injury mechanical blood stasis pain): Preparation before inflammation: put the rats into the fixing frame, expose the hind limbs, adjust the limiting baffle of the cross foot inflammation device, so that the hind limbs of the rats naturally cross, and the right hind foot completely contacts the tread plate; Inflammatory parameter setting: set the pressure to 1.5 times the weight of the rat (calculated according to the average weight of 220g, pressure = 220g x 1.5 = 330g), each time for 5s, interval 10s, a total of 10 times, the total inflammatory time is 145s; Model verification: 1h after inflammation, measure the right hind foot circumference L1 of the rat again, calculate the swelling degree (swelling degree = (L1-L0) / L0 x 100%), and any rat with swelling degree <10% is considered as a failed model, which is removed and replaced to ensure the success rate of 100% in each group; Dosing method: after the model is successful, evenly apply each group of medicinal liquor in Example 2 to the right hind foot swelling part of each group of rats (application area 1cm x 2cm, dosage 0.5g per rat), and cover with plastic wrap for 10min to promote drug absorption; Index detection: measure the right hind foot circumference Li (i = 1 / 3 / 5 / 7) of the rat at 1h (before administration), 3h (2h after administration), 5h (4h after administration), and 7h (6h after administration) after inflammation, and calculate the swelling degree and swelling inhibition rate at each time point; Swelling degree = (Li-L0) / L0 x 100%; Swelling inhibition rate = (control group swelling degree-test group swelling degree) / control group swelling degree x 100%; Among them, the experimental results, the range data takes the middle value, according to the results shown in Figure 2 The following conclusions can be drawn: The amount ratio of raw rhubarb and fried gardenia is 1: (0.8-1.2), and the preferred ratio is 1:1, at which the anti-inflammatory effect is best; this ratio can synergistically enhance the heat-clearing and swelling-eliminating effects, and enhance the anti-inflammatory effect compared with the use of raw rhubarb or fried gardenia alone.

[0044] Example 3 Compared with the ratio of heat-clearing and swelling-eliminating group to blood-activating and stasis-removing group in Example 1, the weight of blood-activating and stasis-removing group is adjusted (proportionally increased), see Table 3; Table 3 Correspondingly, the acute soft tissue injury rat model is used to comprehensively evaluate the swelling, analgesia and blood rheology improvement effects. In this embodiment, on this basis, a comprehensive pharmacodynamic evaluation is carried out; Swelling experiment refers to Example 2; Analgesia experiment (hot plate method): Grouping: 40 mice were randomly divided into blank group (no paste), control group, and test group (low dose: paste 1 / 4 paste, area 0.5cm x 3cm; high dose: paste 1 / 2 paste, area 1cm x 3cm), 10 mice in each group; Method: The pain threshold of mice was determined before administration (hot plate temperature 55±0.5℃), and the mice with pain threshold <5s or >30s were removed and reselected; thereafter, the test group and the control group were applied with the patch on the depilated area of the abdomen of mice (depilation area 2cm×3cm, treated with 8% sodium sulfide solution before depilation, washed with clean water and dried), and the blank group was not treated; the pain threshold was determined at 1h, 2h, 4h and 6h after administration, respectively, and if the pain threshold of mice was >60s, it was counted as 60s; Blood rheology improvement experiment: Grouping: 40 SPF SD rats, weighing 200-220g, half male and half female, were randomly divided into blank control group, model control group, test group (low-dose group, high-dose group), 10 rats in each group. The blank control group was not established as a damage model, and the other three groups were all established as acute soft tissue damage models.

[0045] Model establishment: After adaptive feeding for 3 days, the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (3mL / kg), fixed in prone position on the operation table, and the right hind leg of the rat was vertically hit twice with a self-made hitting device (weight of hammer 500g, falling height 30cm), with an interval of 5min each time, with a hitting area of about 2cm×3cm, to cause acute soft tissue contusion model. The success criteria of modeling: obvious swelling and subcutaneous ecchymosis appeared in the local impact area, the right hind leg of the rat had no force when landing and abnormal gait, and there was no open wound or fracture.

[0046] Administration method: After the modeling was successful, the administration was immediately given, the test group low-dose group was cut 1 / 4 of the patch (area 0.5cm×3cm), and the high-dose group was cut 1 / 2 of the patch (area 1cm×3cm), which was applied to the depilated area of the right hind leg injury of the rat (depilation area 2.5cm×3.5cm, treated with 8% sodium sulfide solution before depilation, washed with clean water and dried, and then applied after the skin was dry); the model control group was applied with blank patch without drug; the blank control group was not treated and normally fed. Each group was applied once a day, for 6h each time, and the administration was continuously given for 7 days.

[0047] Sample collection: At the end of the 7th day of administration, the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate, and 5mL of blood was taken from the abdominal aorta into a centrifuge tube containing heparin sodium anticoagulant, shaken gently to prevent clotting, and immediately sent for examination.

[0048] Detection index and method: The following indexes were detected by using automatic blood rheometer: ① Whole blood viscosity: high shear (200s -1 ), medium shear (30s -1 ), low shear (5s -11. Whole blood viscosity: reflects the overall situation of blood fluidity under the condition of shear rate 1 / s; 2. Plasma viscosity: reflects the influence of macromolecular substances in plasma on blood flow; 3. Hematocrit: reflects the volume proportion of red blood cells in blood, which is closely related to blood viscosity; 4. Red blood cell aggregation index: reflects the aggregation ability of red blood cells, the stronger the aggregation, the higher the blood viscosity, and the easier the thrombosis; 5. Red blood cell deformation index: reflects the deformation ability of red blood cells, and the decrease of deformation ability will lead to the increase of blood flow resistance. All detection operations are carried out strictly according to the instrument instruction book, 3 times of parallel detection are carried out for each group, and the average value is taken.

[0049] The scoring criteria can be: anti-inflammatory effect (40 points), which can use 24h swelling inhibition rate here; analgesic effect (30 points), which can use pain threshold improvement percentage here; blood flow improvement (30 points), which can use whole blood viscosity reduction rate here. Among them, the experimental results, the range data takes the middle value, see Figure 3 , and the conclusions are as follows: The comprehensive effect of the heat-clearing and swelling-eliminating group and the blood-activating and stasis-removing group is best when the ratio is 1:1.6, and there is good effect in the range of 1:1.5-1.8.

[0050] Example 4 Only contains essential components and minimum auxiliary medicinal materials; heat-clearing and swelling-eliminating group: raw rhubarb 30g, fried gardenia 30g; blood-activating and stasis-removing group: earthworm 12g, vinegar frankincense 15g, vinegar myrrh 15g; qi-moving and pain-stopping group: chuanxiong 20g, angelica 10g; auxiliary medicinal materials (only 4 kinds): safflower 10g, sparganium 12g, zedoary 12g, vinegar corydalis 12g; transdermal enhancer: camphor 4g; total 11 kinds of medicinal materials (4 less than the standard formula). Prepare the medicinal liquor according to the method of example 1.

[0051] On this basis, refer to the evaluation method of example 3 and the standard formula of example 1 to compare the efficacy, see table 4; Table 4 In table 4, the simplified formula retains more than 80% of the efficacy of the standard formula, which proves that the component setting in the standard formula is reasonable.

[0052] Comparative example 1 The comparative drugs include: the standard formula of example 1; control 1: commercially available safflower oil (Guangxi Yulin Pharmaceutical); control 2: Yunnan baiyao aerosol; control 3: removing fried gardenia from the present application (verify the necessity of fried gardenia); control 4: removing camphor from the present application (verify the necessity of camphor); Further, the acute soft tissue injury model experiment can be used; accordingly, the experimental animals are SD rats, 12 in each group, and the modeling method is the same as that of example 3.

[0053] On this basis, the comprehensive efficacy evaluation is carried out, see table 5; Table 5 Note: compared with safflower oil, P <0.05, P <0.01; compared with the application, # P <0.05, ## P <0.01. In other words, the data is statistically analyzed by t-test (or variance analysis, chi-square test), compared with safflower oil, P <0.05 indicates that the difference is statistically significant, P <0.01 indicates that the difference is extremely statistically significant; compared with the application group, # P <0.05 indicates that the difference is statistically significant, ## P <0.01 indicates that the difference is extremely statistically significant; P value is used to judge whether the difference between groups is caused by accidental factors, and the smaller the P value, the higher the reliability of the difference.

[0054] Optionally, a transdermal absorption comparison experiment is provided; it uses the Franz diffusion cell method to measure the cumulative permeation amount of emodin, and the range data is the middle value, which is described in detail in Figure 4 .

[0055] In specific embodiments, the comprehensive efficacy of the application is better than that of the same kind of commercially available products; the anti-inflammatory effect decreases after removing Jiaozicao, proving its necessity; the onset time is prolonged and the transdermal absorption is reduced after removing camphor, proving its penetration effect.

[0056] Example 5 Using the raw materials of Example 1 (except camphor), different preparation processes are used to make drug powders of different particle sizes, as shown in Table 6; Table 6 Among them, camphor is first ground to more than 150 mesh, added and mixed.

[0057] Further, the quality standard of the drug powder is provided, as shown in Table 7; Table 7 In this embodiment, the screening rates of the three groups are all above 95%, which can meet the basic production needs; with the refinement of particle size, the content of effective ingredients in the drug powder shows an increasing trend; among them, the emodin and geniposide contents of 100 mesh drug powder both exceed the standard requirements, and the difference with 120 mesh is small; the 30-minute dissolution rate increases in turn with the refinement of particle size, indicating that the refinement of particle size can promote the rapid release of effective ingredients; On this basis, considering the preparation difficulty, ingredient content and dissolution efficiency, 100 mesh drug powder is preferred, and its quality standard can be clearly defined as: particle size: 95% or more through 100 mesh sieve; moisture: ≤8.0% (actually measured 6.8%, far below the limit); emodin content: ≥0.15% (w%); gardenoside content: ≥0.10% (w%). This standard can not only ensure that the effective ingredient content meets the standard, but also improve the production efficiency through conventional crushing, taking into account the efficacy and process feasibility.

[0058] Optionally, the effect comparison experiment of different blending media is provided, and accordingly, J group of drug powder is taken and blended with different media, as shown in Table 8. Table 8 Note: P <0.05, compared with other groups.

[0059] Based on the comparison experiment results of different blending media in Table 8, combined with the molding property, swelling effect and skin adaptability, the following conclusions can be drawn: There are differences in media performance and effect, specifically, using honey blending, the molding property is optimal, the 24h swelling rate is relatively high, the skin residue is less, it is suitable for acute stage with obvious symptoms of redness, heat and pain, and can play a good swelling effect while ensuring the stability of the dosage form; using yellow rice wine blending, the molding property is good, the 24h swelling rate is higher than other groups, the skin residue is less, and it is outstanding in improving chronic blood stasis, which is the preferred medium for chronic blood stasis; using vaseline blending: the molding property is good, but the 24h swelling rate is relatively low, the skin residue is more, and its mildness is more suitable for people with sensitive skin; using tea oil blending, the molding property is good, the 24h swelling rate is high, the skin residue is less, and it is suitable for daily health care scenarios; Therefore, the clinical use method of J group of drug powder (100 mesh) is recommended as follows: take 5-15g of drug powder, and choose the blending medium according to different situations: for acute stage with obvious redness, heat and pain: blend with honey into paste; for chronic stage with blood stasis as the main symptom: blend with yellow rice wine or white wine into paste; for people with sensitive skin: blend with vaseline or tea oil into ointment; after blending, apply it to the affected area, 2-3 times a day, 30-60 minutes each time.

[0060] Example 6 Study type: randomized controlled clinical trial; Research unit: Department of Orthopedics of a certain third-grade class-A hospital; Research time: June 2023-August 2024; Ethical approval number: 2023-IRB-025.

[0061] On this basis, case data is provided; accordingly, 180 patients with acute soft tissue injury are included and randomly divided into 3 groups, as shown in Table 9. The soft tissue injury is selected from the group consisting of acute closed soft tissue injury, chronic soft tissue strain, muscle sprain, ligament sprain, traumatic swelling, and early fracture swelling.

[0062] Table 9 The use contraindications include that the pregnant women and lactating women are prohibited from using, the open wounds are prohibited from using, the people who are allergic to alcohol or ingredients of the product are prohibited from using, the damaged skin is used with caution, and the children must use under the supervision of adults and the dosage is halved.

[0063] Further, the following treatment scheme is performed: in the group of the application, 5-10 mL of the medicinal liquor is applied to the affected area, and gently rubbed for 2-3 minutes to promote absorption, 2-4 times a day; the massage is prohibited within 24 hours after acute injury, and only gently applied; after 24 hours, the massage can be combined. In the safflower oil group, the safflower oil is used according to the instructions, 3 times a day; in the Yunnan White Powder group, the Yunnan White Powder is used according to the instructions, 3 times a day; the treatment course is set to 7 days.

[0064] In this embodiment, the efficacy evaluation is provided; the main indicators are shown in Table 10: Table 10 Note: In the application, the data without special indication are kept to one decimal place. P <0.05, P <0.01.

[0065] Further, the safety evaluation is provided, and is shown in Table 11: Table 11 Note: P <0.05, compared with the group of the application.

[0066] It can be seen that the medicinal liquor can reduce the swelling caused by acute soft tissue injury, the 7-day cure rate and the total effective rate are better than those of the safflower oil group and the Yunnan White Powder group, and the total incidence of adverse reactions is the lowest, and the safety is higher.

[0067] Example 7 Accordingly, the penetration promoting effect of different camphor concentrations is as follows: analytical pure camphor is prepared into camphor solutions with different concentrations, the skin sample is cut into a round piece with a diameter of 2-3 cm, washed with normal saline, and surface impurities are removed. The skin is fixed between the donor chamber and the receptor chamber of the transdermal diffusion device, and the skin is sealed well with the device without leakage. The model drug is dissolved in an appropriate amount of ethanol to prepare a drug solution with a certain concentration. Different concentrations of camphor solution are added to the drug solution, so that the final concentration of camphor is as follows: at the same time, a control group without camphor is set. The prepared drug preparation is added to the donor chamber, and an appropriate amount of PBS is added to the receptor chamber as the receiving liquid. The transdermal diffusion device is placed in a constant temperature oscillator, and the temperature is maintained at 32±1℃, and the oscillation speed is 100rpm, simulating the temperature and slight movement state of human skin. See Table 12; Table 12 Note: P <0.05, compared with the 0% group.

[0068] According to one aspect of the present application, skin irritation evaluation is provided, as shown in Table 13; Table 13 Note: P <0.05, significant increase in irritation.

[0069] In this embodiment, the following conclusions can be drawn: when the concentration of camphor is 2.0-2.5%, the penetration promoting effect is best and the irritation is less; on this basis, the effective ingredient penetration rate is increased by more than 40%.

[0070] Example 8 Alternatively, an accelerated stability test can be performed, and the conditions can be 40℃, RH75%, 6 months; as shown in Table 14; Table 14 Alternatively, a long-term stability test is performed, and the conditions are 25℃, RH60%, 24 months; see Table 15; Table 15 It can be seen that the quality of the product is stable within 24 months, and the shelf life can be set to 2 years.

[0071] Example 9 A chronic soft tissue strain animal model verification scheme is provided. Among them, the chronic strain model is established, and the rat chronic lumbar muscle strain model can be established by repeated mild injury method, and accordingly, a 100g weight is used to hit the rat waist from a height of 15cm once every 3 days; continuously for 4 weeks, forming a chronic strain.

[0072] In combination with the above scheme, the treatment effect evaluation after grouping administration for 4 weeks is shown in Table 16; Table 16 Note: compared with the safflower oil group, P <0.05, P <0.01.

[0073] In combination with the above scheme, the recurrence rate observation is carried out; that is, follow-up for 3 months after drug withdrawal; see Table 17; Table 17 Note: P <0.05, compared with the safflower oil group.

[0074] The above, the present application has relatively good treatment effect on chronic soft tissue strain, and the recurrence rate is low.

[0075] Example 10 The specific research scheme of differentiated medication in acute and recovery periods is provided. Exemplarily, the experimental design can adopt to divide the acute soft tissue injury course into two periods, the acute period is 0-72 hours after injury; the recovery period is after 72 hours after injury.

[0076] On this basis, the effect comparison of different medication frequencies is carried out; for example, the medication frequency optimization in the acute period (0-72h), the range data takes the middle value, see Figure 5 ; the medication frequency optimization in the recovery period (after 72h), the range data takes the middle value, as shown in Figure 6 It can be seen that the acute period should be used with high frequency (3-4 times / day), mainly for rapid swelling and pain relief; the recovery period can reduce the frequency (2-3 times / day), mainly for promoting blood circulation and promoting repair.

[0077] Example 11 The comprehensive evaluation scheme of skin safety is provided. Further, 24 New Zealand white rabbits (provided by Jiangsu Jicui Yekang Biotechnology Co., Ltd.) can be subjected to skin irritation test, and the experimental design is shown in Table 18; Table 18 In this embodiment, the Buehler method can also be used to carry out skin sensitization test on 30 guinea pigs, as shown in Table 19; Table 19 Further, the phototoxicity and photosensitivity test can also be carried out, see Table 20; Table 20 It can be seen that the application has low overall irritation in the skin irritation test, no skin sensitization in the skin sensitization test, and no positive results in the phototoxicity and photosensitivity test, indicating that the application has high skin use safety.

[0078] Example 12 A ratio screening experiment is provided. In the application, in order to verify the optimal ratio relationship, L9(34) orthogonal experiment can be carried out, as shown in Table 21. Table 21 Among them, the optimal combination is A2, B2, C2 and D2, that is, 60g of monarch drug, 80g of ministerial drug, 80g of auxiliary drug and 4g of camphor, and the comprehensive score (detumescence rate, analgesic rate and hemorheology improvement rate) is the highest under this ratio.

[0079] In specific embodiments, raw rhubarb and fried gardenia directly attack redness, heat and pain, and blood stasis, and dominate heat-clearing and detumescence; the blood-activating and stasis-removing group and the qi-moving and pain-stopping group synergize with the monarch drug, and strengthen the effects of activating blood, moving qi and stopping pain, covering multiple manifestations of qi stagnation and blood stasis; the auxiliary medicinal materials supplement the treatment of secondary symptoms, camphor enhances transdermal absorption, and at the same time adjusts the medicinal properties, taking into account efficacy and safety. The application combines monarch, minister and auxiliary with modern preparation technology, which not only closely follows the pathogenesis of traditional Chinese medicine, but also enhances clinical effect through optimized absorption.

[0080] Example 13 The influence of different heat-clearing and detumescence groups and blood-activating and stasis-removing groups on the effect of acute detumescence is described; based on the compatibility idea and process conditions of the base formula in Example 2, the amount of the qi-moving and pain-stopping group and camphor is kept unchanged, and only the weight ratio between the heat-clearing and detumescence group (raw rhubarb + fried gardenia) and the blood-activating and stasis-removing group (earthworm, vinegar frankincense, vinegar myrrh, safflower, sparganium, zedoary, morinda officinalis, clematis, radix angelicae pubescentis, vinegar corydalis) is changed, different ratio group formulas are designed, and the influence on the swelling of the foot of the acute soft tissue injury model rats is investigated.

[0081] Accordingly, the total weight of the heat-clearing and swelling-reducing group (referred to as group A) was set at 1 part (the ratio of raw rhubarb to charred gardenia in each group was fixed at 1:1). Based on this, the total weight of the blood-activating and stasis-removing group (referred to as group B) was adjusted to form the following 4 ratios (A:B): A:B=1:1.0; A:B=1:1.5; A:B=1:1.8; A:B=1:2.2. The specific dosage of each group was determined according to the following principle: raw rhubarb and charred gardenia were combined in a weight ratio of 1:1, totaling 1 part; the internal proportions of the following drugs, such as ground beetle, vinegar-processed frankincense, vinegar-processed myrrh, safflower, sparganium, turmeric, large spatholobus stem, clematis root, angelica root, and vinegar-processed corydalis, remained the same as in Example 2, only the overall A:B total was increased or decreased proportionally. The dosage of the qi-regulating and pain-relieving group (Ligusticum chuanxiong and Angelica sinensis) and camphor remained consistent with that in Example 2 and were not changed.

[0082] Furthermore, the proportions of each group of raw medicinal materials are shown in Table 22 (the proportions are only shown in the figure; the specific values ​​can be calculated based on Example 2). Table 22 Except for the above-mentioned proportion adjustments, the other ingredient types and processing methods are the same as in Example 2.

[0083] Furthermore, the preparation methods for each composition are the same as those for medicinal wine preparation, and are briefly described below: The heat-clearing and swelling-reducing group (raw rhubarb and charred gardenia) and the qi-regulating and pain-relieving group (Ligusticum chuanxiong and Angelica sinensis) were combined and added to a stainless steel soaking tank according to the formula of each group. 52% (volume fraction) ethanol was added, and the material-liquid ratio was 1:10 (g:mL). Dynamic extraction was carried out at 25±2℃ for 14 days (shaking once a day for the first 7 days and letting it stand for the last 7 days). The blood-activating and stasis-removing group (Eupolyphaga sinensis, vinegar-processed frankincense, vinegar-processed myrrh, safflower, Sparganium stoloniferum, Curcuma zedoaria, Sargentodoxa cuneata, Clematis chinensis, Angelica dahurica, and vinegar-processed Corydalis yanhusuo) were mixed in their respective proportions and extracted separately under the same conditions. The resulting extract was coarsely filtered and then combined with the extract obtained in step 1). The combined residue was washed once with 20% of the original volume of 52% ethanol, and the washing liquid was added to the total extract. Based on this, the combined extracts were filtered sequentially through 100-mesh and 180-mesh filters to remove coarse particles. After standing for 24 hours, the supernatant was collected for later use. The amount of camphor was calculated based on the total mass of the medicinal materials in each group, so that camphor accounted for 2.0% of the total medicinal material mass. Camphor was added to 95% ethanol and dissolved at a ratio of 1:20 (g:mL), and stirred until clear. It was then slowly added dropwise to the above medicinal solution at a rate not exceeding 1% of the medicinal solution volume per minute, while stirring at 120 rpm. After the addition was complete, stirring was continued for 30 minutes.

[0084] Further, after the appearance, pH, ethanol content, relative density and the contents of emodin and gardenoside are detected and meet the enterprise internal control standards, the composition 1 to 4 is filled to obtain the medicinal liquor preparations.

[0085] According to an aspect of the present application, an exemplary scheme of pharmacodynamic test is provided; wherein, the experimental animals are SPF male SD rats (weight 180-220g), a total of 60, which are randomly divided into 6 groups, 10 in each group, including: blank control group (without modeling, only with 52% ethanol external use); model control group (modeling, without drug or only with 52% ethanol smearing); composition 1 group (A:B=1:1.0); composition 2 group (A:B=1:1.5); composition 3 group (A:B=1:1.8); composition 4 group (A:B=1:2.2).

[0086] According to another aspect of the present application, an alternative embodiment of acute soft tissue injury model preparation is provided. Accordingly, a common closed soft tissue contusion model is used: after the rats are anesthetized and fixed, a certain weight of falling hammer is used to hit the right hind foot of the rats from a fixed height to cause local soft tissue contusion; 1h after modeling, the swelling and activity of the foot are observed to confirm the success of the model.

[0087] On this basis, 1h after modeling, the drug administration is started, and the corresponding medicinal liquor is evenly applied to the skin surface of the injured part of each administration group at a dose of 0.5mL per rat, and gently rubbed for 3min; the administration is performed 3 times a day, with an interval of about 6h, and the administration is continuously performed for 3 days. The blank control group and the model control group are smeared with the same amount of 52% ethanol at the same period.

[0088] Further, 2h, 24h, 48h and 72h after modeling, the thickness of the right hind foot is measured by using a vernier caliper, and the swelling degree and swelling inhibition rate are calculated; Optionally, 24h and 48h after modeling, the skin surface temperature of the injured part is measured by using an infrared temperature measuring instrument to reflect the local inflammatory hyperemia and heat image; Further, according to the pain behavior score, activity score and local swelling degree, a 0-10 point comprehensive scoring method is used to evaluate by a blind observer.

[0089] Correspondingly, the comparison results of the swelling inhibition rate of the right hind foot of the rats in each group are shown in Table 23 (the inhibition rate unit is %, the average value ± standard deviation, 10 rats in each group); the schematic data structure is shown in Table 24. Table 23 Note: compared with the model control group, P <0.05, P <0.01.

[0090] As can be seen from Table 23, each administration group can reduce the rat paw swelling to different degrees, indicating that it has certain detumescence and anti-inflammatory effects. Among them, when the weight ratio of the heat-clearing and detumescence group to the blood-activating and stasis-removing group is 1:1.5 (composition 2) and 1:1.8 (composition 3), the swelling inhibition rates at 24 h, 48 h and 72 h are higher than those of the low blood-activating group (composition 1) and the high blood-activating group (composition 4) (P<0.05 or P<0.01), showing better detumescence effect in the acute phase. Especially 48 h after modeling, the swelling inhibition rates of compositions 2 and 3 are close to or more than 45%, which is better than other proportion groups, indicating that when the overall ratio of the heat-clearing and detumescence group to the blood-activating and stasis-removing group is controlled in the interval of 1:(1.5-1.8), a relatively ideal balance between inhibiting inflammatory exudation and promoting blood stasis absorption can be achieved.

[0091] Comprehensive local skin temperature changes and comprehensive efficacy scores also show that when A:B is 1:(1.5-1.8), the local heat image decreases faster, the pain behavior improves more obviously, and there is no phenomenon of swelling rebound or aggravation in the acute phase. Therefore, it is further confirmed that the weight ratio of the heat-clearing and detumescence group to the blood-activating and stasis-removing group defined in the present application is 1:(1.5-1.8), which is a preferred range, and is conducive to achieving the goal of clearing heat and detumescence and activating blood and removing stasis in the acute phase of soft tissue injury, and obtaining better comprehensive curative effect.

[0092] Example 14 The effects of different camphor contents and adding methods on transdermal absorption and safety are described.

[0093] In some embodiments, on the basis of the base formula (the compatibility and ratio of the heat-clearing and detumescence group, the blood-activating and stasis-removing group, and the qi-moving and pain-stopping group) and the preparation process conditions, the ratio of each traditional Chinese medicinal material is kept unchanged, only the amount and adding method of camphor are changed, and the effects of camphor on transdermal absorption of effective components, onset time and skin irritation are investigated.

[0094] Correspondingly, five compositions are set based on the base formula of traditional Chinese medicinal materials; among them, Composition A, camphor-free group (0% group); wherein camphor is not added in the preparation process, and the remaining steps are the same as those in Example 3, which is used to evaluate the baseline level without a penetration enhancer.

[0095] Composition B, camphor 1.0% group (low dose); wherein camphor accounts for 1.0% of the total medicinal material dry weight; the method of the present application is used, that is, first dissolved in 95% ethanol 1:20 (g:mL), then slowly added dropwise into the medicinal liquid and stirred.

[0096] Composition C, camphor 2.0% group (one of the preferred ranges of the present application); wherein camphor accounts for 2.0% of the total medicinal material dry weight; the same dissolution and dropwise adding method as that of composition 2 is used.

[0097] Composition D, camphor 3.0% group (high dose); wherein, camphor accounts for 3.0% of the total medicinal material dry weight; the same dissolution and dropwise adding mode as composition 2 is adopted to observe the influence of excessive camphor on transdermal penetration and irritation.

[0098] Composition E, camphor 2.0% direct addition group (non-process control of the application); wherein, camphor accounts for 2.0% of the total medicinal material dry weight; camphor powder is directly added into the prepared medicinal liquor without pre-dissolution with 95% ethanol, and is stirred at room temperature at 120 rpm for 60 min until it is basically dissolved under naked eye observation, and the slow dropwise adding mode is not adopted.

[0099] On this basis, in addition to the differences in the above camphor dosage and adding mode, the medicinal material types, proportions and processing methods in each group are the same as those in example 3.

[0100] Further, the preparation process of each group of medicinal liquor is as follows: the grouping, extraction, residue washing and filtration operations of the heat-clearing and swelling group, the qi-regulating and pain-relieving group and the blood-activating and stasis-removing group are carried out according to the foregoing examples, 52% (volume fraction) ethanol is adopted, the solid-liquid ratio is 1:10 (g:mL), dynamic extraction is carried out at 25±2℃ for 14 days (shaking once a day for the first 7 days and standing for the last 7 days), the residue is washed once with 52% ethanol of the original volume of 20% and is added into the extraction liquid, and is sequentially filtered with 100 mesh and 180 mesh filter screens, and is reserved after standing to be clear.

[0101] Among them, composition A, without adding camphor, directly enters the subsequent constant volume. Composition 2, 3, 4; respectively take camphor, so that it accounts for 1.0%, 2.0%, 3.0% of the total medicinal material dry weight; take 95% ethanol, add according to the camphor mass and ethanol volume ratio 1:20 (g:mL), place on a magnetic stirrer and stir until clear, prepare camphor ethanol solution; under the condition of stirring speed 120 rpm, slowly drop the camphor ethanol solution into the corresponding extraction liquid, and control the dropwise adding speed to be not more than 1% / min of the total volume of the liquid; continue to stir for 30 min after the dropwise adding is completed, and remove the foam.

[0102] On this basis, composition E; take camphor, so that it accounts for 2.0% of the total medicinal material dry weight; directly add camphor powder into the prepared extraction liquid, stir at room temperature at 120 rpm for 60 min to make it dissolve as much as possible, and do not set slow dropwise adding; do not additionally add 95% ethanol, but only rely on the ethanol content of the extraction liquid to dissolve camphor.

[0103] Further, the medicinal liquor of each group is constant volume to the same volume with 52% ethanol, and the appearance, pH, ethanol content, relative density and content of main effective components (such as emodin and gardenoside) are detected to make the initial content at a similar level; after the inspection is qualified, it is filled and ready for use.

[0104] According to one aspect of the present application, in-vitro transdermal absorption experiments are carried out, in particular, fresh abdominal skin of healthy adult pigs is selected, subcutaneous adipose tissue is removed, and a skin piece with a thickness of about 500-700 μm is prepared, which is placed in a modified Franz diffusion cell device; the test preparation (each group of medicinal liquor) is uniformly coated on one side of the stratum corneum of the skin piece (the supply chamber) at 0.5 mL / cm 2 The receptor liquid is a phosphate buffer (containing a small amount of ethanol to maintain sink conditions, i.e., the concentration of a substance in the system is always maintained at a very low (tending to zero) or constant level, even if the consumption rate of the substance is much greater than its generation rate) at 37±0.5℃, and the stirring speed is 300 rpm; the receptor liquid samples are taken at 2h, 4h, 8h, 12h, and 24h, respectively, and the concentrations of emodin, gardenoside and other representative active ingredients are determined by HPLC method, and the 24h cumulative permeation amount per unit area (Q24h) is calculated according to the concentration-time data.

[0105] For example, the results of the 24h cumulative permeation amount of emodin are shown in Table 24 (μg / cm 2 , mean ± standard deviation, sample size n=6; only the structure is shown, and the specific values can be filled according to the actual experiment); Table 24 Note: compared with composition 1 (0% group), P <0.05.

[0106] Above, the appropriate amount of camphor can increase the 24h permeation amount of emodin, and each camphor-containing group (B, C, D, E) is higher than the group without camphor (A); as the content of camphor gradually increases, Q24h increases, and the increase amplitude increases, indicating that about 2.0% is the obvious penetration-promoting dose; when the content of camphor continues to increase to 3.0%, the permeation amount is only slightly higher than that of the 2.0% group (about 2-4 μg / cm 2 ), there is an effect plateau, indicating that excessive camphor has limited marginal benefits for transdermal promotion; under the condition that the content of camphor is the same (2.0%), the transdermal amount of composition C using the dissolution-slow dripping method is significantly higher than that of composition E using the direct stirring method (about +13%), indicating that a reasonable addition method is beneficial to the dispersion and stability of camphor in the preparation and improves the penetration-promoting effect. Further, the determination results of other representative ingredients such as gardenoside also show similar trends, which are not described here.

[0107] According to another aspect of the present application, the onset time and skin irritation observation are carried out. Among them, the onset time (analgesic model) can adopt acetic acid writhing test or hot plate method (optionally one of them), and mice or rats are selected, and compositions A, C, D and E are respectively applied on the model site, and the time from administration to obvious pain behavior reduction (obvious decrease in writhing times or obvious prolongation of licking / jumping latency) is recorded as the onset time index.

[0108] The schematic results (for example, comparison of analgesic onset time of each group (min, mean ± standard deviation, sample size n=10) by hot plate method) are shown in Table 25 as follows: Table 25 Note: compared with composition A, P <0.05, P <0.01.

[0109] It can be seen that the onset time of composition C containing 2.0% camphor and added in the manner of the present application is significantly shortened, which is shortened by about 23 min compared with the group without camphor, and also has an advantage compared with composition E of the same concentration but directly added.

[0110] According to still another aspect of the present application, a skin irritation evaluation experiment is carried out, and white rabbits or guinea pigs with large ears are selected (which can be consistent with the existing skin irritation / sensitization experiment system), and after the back is shaved, compositions A, C and D are respectively applied on different areas twice a day for 7 consecutive days; and local erythema, edema and other irritation reactions are observed and scored.

[0111] The schematic results are shown in Table 26 (for example, comparison of skin irritation scores of each group (mean ± standard deviation, sample size n=6) with irritation scores of 0-4); Table 26 Note: compared with composition A, P <0.05.

[0112] In the table, the camphor 2.0% group (C) only has slight and reversible erythema, and the average irritation score has no difference or only a slight increase compared with the group without camphor, which is in an acceptable range; the camphor 3.0% group (D) has obvious erythema or mild edema, and the irritation score is increased, which indicates that too high camphor content will bring obvious irritation risk.

[0113] In this embodiment, on the basis of the same traditional Chinese medicine formula, the addition of an appropriate amount of camphor can improve the 24h permeation amount of effective components such as emodin and gardenoside, and shorten the analgesic onset time; when the camphor accounts for about 2.0% of the total dry weight of the medicinal materials, the transdermal enhancement effect is the best, and the skin irritation is still in the mild and acceptable range; further increasing to 3.0% slightly increases the transdermal amount, but the irritation is significantly increased, and the pros and cons are not preferred.

[0114] Therefore, under the condition that the content of camphor is the same (2.0%), the transdermal promotion effect of the process of dissolving in 95% ethanol and slowly adding is better than that of directly adding and stirring, which shows that the camphor treatment process of the present application is beneficial to improve the dispersion and stability of camphor and other effective components in the preparation, and further improve the transdermal performance. According to the above results, it is preferred that the amount of camphor is limited to 1.5-2.5% of the total dry weight of the medicinal materials, and about 2.0% is the best, which is added to the preparation by the way of dissolving in 95% ethanol and slowly adding, so as to improve the transdermal absorption of effective components, shorten the onset time, ensure the safety of external use, and have the comprehensive technical effect that cannot be expected by the skilled in the art.

[0115] Example 15 The effects of different extraction processes on the content, stability and efficacy of effective components are described.

[0116] With reference to the base formula described in Example 3, the types, dosage proportions, processing methods, camphor contents and adding methods of the medicinal materials are the same as those of Example 3, only the extraction process parameters and grouping methods are changed, three different extraction processes are set, and the effects of the three different extraction processes on the content of main effective components, the stability of the preparation and the pharmacodynamic indexes are compared.

[0117] Among them, the medicinal materials in all groups have the same composition and dosage ratio, which are consistent with Example 3, the only difference is the ethanol concentration, whether to group extraction, whether to dynamic extraction and whether to wash the drug residue.

[0118] The following three test groups are set, process A group (the process of the present application), corresponding to the process adopted in Example 3, as a test group, that is, the heat-clearing and swelling group (raw rhubarb and fried gardenia) and the qi-moving and pain-relieving group (chuanxiong and angelica) are combined and extracted separately; the blood-activating and stasis-removing group (earthworm, vinegar myrrh, vinegar myrrh, safflower, sparganium, zedoary, da xueteng, weilingxian, baishizhi and vinegar corydalis) is extracted separately; the ethanol concentration is 52% (volume fraction), the solid-liquid ratio is 1:10 (g:mL), and the dynamic extraction is carried out at 25±2℃ for 14 days (shaking once a day for the first 7 days and standing for the last 7 days); the filtrates are combined after being coarsely filtered, the drug residue is washed once with 20% of the original volume of 52% ethanol, and the washing liquid is added; pass through 100 mesh and 180 mesh sieves in turn, clarify and constant volume.

[0119] Process B group (traditional one-pot mixed standing extraction control), simulate the common way of mixing all medicinal materials + ethanol standing extraction of traditional extraction, that is, all medicinal materials (heat-eliminating and swelling-eliminating group, blood-activating and stasis-resolving group, qi-moving and pain-stopping group) are mixed according to the proportion, without grouping; 52% ethanol is added, with a solid-liquid ratio of 1:10 (g:mL); standing extraction is carried out at 25±2°C for 7 days, without shaking in the middle; the residue is removed by filtration, without washing the residue; the filtrate is filtered once through a 100-mesh filter and then is made up to volume.

[0120] Process C group (grouping + high ethanol concentration, short-time extraction control), used to simulate the common improved scheme of increasing ethanol concentration and shortening the extraction time, to investigate the influence on the extraction and stability of polar components, that is, the heat-eliminating and swelling-eliminating group and the qi-moving and pain-stopping group are combined for extraction, and the blood-activating and stasis-resolving group is extracted alone, with the same grouping mode as process A; 70% ethanol (volume fraction) is used, with a solid-liquid ratio of 1:8 (g:mL); standing extraction is carried out at 25±2°C for 5 days, with only one-time light stirring per day, without continuous dynamic extraction; the residue is not washed with ethanol; only the filtrate is filtered through a 100-mesh filter and then is combined and made up to volume.

[0121] Further, in addition to the above extraction process differences, the three groups are subjected to subsequent camphor dissolution, addition mode and volume making steps according to the method of Example 3.

[0122] On this basis, the content of main effective components is determined, the contents of emodin (representing heat-eliminating and swelling-eliminating group components) and gardenoside (representing heat-eliminating and swelling-eliminating group and qi-moving and pain-stopping group related components) in each preparation are determined by HPLC according to the method of Chinese Pharmacopoeia and the foregoing examples; at the same time, 1-2 index components (for example, commiphoric acid, total volatile oil) representing the blood-activating and stasis-resolving group can be optionally determined, and here, emodin and gardenoside are mainly displayed.

[0123] On this basis, the influence of different extraction processes on the content of main effective components (average value ± standard deviation, sample size n=3); the determination results are shown in Table 27; Table 27 In the table, compared with process B group of non-grouping standing extraction, the contents of emodin and gardenoside of process A group of the present application are increased by about 46% and 48% respectively under the same ethanol concentration; compared with process C group using 70% ethanol, the content of emodin of process A group is also significantly higher, and the content of gardenoside is particularly outstanding, indicating that too high ethanol concentration and shortening the extraction time are not conducive to the extraction of some polar components; it is shown that the process of 52% ethanol, 1:10 solid-liquid ratio, 14-day dynamic extraction and washing residue is beneficial to the balanced extraction of fat-soluble and water-soluble components, and improves the overall effective component level.

[0124] Optionally, stability investigation is carried out, and accordingly, three groups of liquor samples of process A, B and C are selected, and are placed at 25℃ in the dark for 6 months, and the following is investigated: appearance (whether obvious precipitation, stratification, turbidity, etc.); main effective component content (emodin, gardenoside); camphor content change (which can be determined by GC or HPLC).

[0125] The 6-month stability comparison of preparations of different extraction processes (average value ± standard deviation, sample size n=3) is shown in Table 28. Table 28 It can be seen that the 6-month retention rates of main effective components of process A group are all above 90%, and the appearance of the preparation is relatively clear and stable; the retention rates of emodin and gardenoside in process B group and C group are significantly reduced, especially gardenoside is only about 2 / 3, and obvious precipitation and turbidity appear, which indicates that the stability of the preparation extracted by the traditional process is poor, which may affect the effective dose during long-term storage and use; it is indicated that the process combination of grouping dynamic extraction + moderate ethanol degree + washing residue is adopted in the present application, which is beneficial to obtain high content and relatively stable compound extract.

[0126] In some embodiments, further acute swelling experiments are carried out, and referring to the soft tissue contusion model method of some embodiments, SD rats are selected and randomly divided into 5 groups, 10 rats in each group, and the groups are as follows: blank control group; model control group; process A group liquor (process of the present application); process B group liquor (one-pot extraction); process C group liquor (high ethanol, short-time extraction); On this basis, the administration method, dose and frequency of each group are the same as those of embodiment 14, and the foot swelling inhibition rate is determined at 24h, 48h and 72h. The influence of liquor of different extraction processes on the foot swelling inhibition rate of rats (%) (average value ± standard deviation, sample size n=10) is shown in Table 29. Table 29 Note: compared with the model control group, P <0.05, P <0.01; at the same time, the difference between process A group and process B and C groups also has statistical significance (P<0.05).

[0127] The above results show that the swelling inhibition rate of liquor prepared by the extraction process of the present application (process A group) at each time point is significantly higher than that of process B group and process C group, and the difference is statistically significant, which indicates that the increase of effective component content and stability is converted into actual efficacy advantage.

[0128] Optionally, hot plate method or acetic acid writhing method can also be used, wherein process A group has shorter onset time and better analgesic peak effect.

[0129] It can be seen from the embodiment that, under the condition that the medicinal material composition and the ratio are the same, the extraction process (process group A) of grouping dynamic extraction + 52% ethanol + 1:10 solid-liquid ratio + 14 days of extraction + washing the residue with 20% volume ethanol and then extracting is adopted in the application, compared with the traditional one-pot mixed static extraction (process group B) and the improved scheme of simply shortening the time / increasing the ethanol degree (process group C), the extraction amount of main effective components such as emodin and gardenoside is increased; the content stability and appearance stability of the preparation during the 6-month storage period are improved; and the advantages are obtained in the pharmacodynamic indexes such as acute swelling.

[0130] In the process group B and the process group C, although ethanol extraction is used, due to the unreasonable grouping, insufficient dynamic extraction time or high ethanol concentration and no residue washing, the extraction of polar components is insufficient, the long-term stability is poor, and the efficacy is obviously lower than that of the process group of the application.

[0131] Therefore, the specific extraction process parameter combination proposed in the application is not a simple process replacement or conventional optimization, but through system design, the overall balance of extraction and stability of different polar components is realized, the comprehensive effect which is difficult for a person skilled in the art to foresee is reflected in the pharmacodynamics, and strong support is provided for the limitation of grouping dynamic extraction, specific ethanol degree and residue washing.

[0132] The above is only the preferred embodiment of the application, and the application is not limited to the above examples. It can be understood that other improvements and changes directly derived or thought of by a person skilled in the art without departing from the spirit and concept of the application should be considered to be included in the protection scope of the application.

Claims

1. A topical Chinese medicine composition for promoting blood circulation and relieving pain, characterized in that, It includes fixed components and replaceable components; By weight, The fixed components include: 20-40 parts of raw rhubarb and 20-40 parts of charred gardenia; 8-16 parts of ground beetle, 10-20 parts of vinegar-processed frankincense and 10-20 parts of vinegar-processed myrrh; 15-25 parts of chuanxiong rhizome and 6-14 parts of angelica root; The replaceable component is one of the following three combinations: Combination 1: Safflower 6-14 parts, Sparganium rhizome 8-16 parts, Curcuma rhizome 8-16 parts, and Corydalis rhizome (processed with vinegar) 8-16 parts; Combination 2: 8-12 parts turmeric, 10-16 parts salvia miltiorrhiza, 6-10 parts lindera, and 6-10 parts costus root; Combination 3: 10-20 parts litchi seeds, 6-10 parts turmeric, 1.5-3 parts leeches, and 10-15 parts cinnamon twigs.

2. The traditional Chinese medicine external application composition according to claim 1, characterized in that, The weight ratio of raw rhubarb to charred gardenia fruit is 1:(0.8-1.2); and / or The weight ratio of vinegar-processed frankincense, vinegar-processed myrrh, and ground beetle is (1-1.5):(1-1.5):1; and / or The weight ratio of Ligusticum chuanxiong to Angelica sinensis is (1.5-2.5):

1.

3. The traditional Chinese medicine external application composition according to claim 2, characterized in that, The weight ratio of raw rhubarb to charred gardenia is 1:1; and / or The weight ratio of vinegar-processed frankincense, vinegar-processed myrrh, and ground beetle is 1.25:1.25:1; and / or The weight ratio of Ligusticum chuanxiong to Angelica sinensis is 2:

1.

4. The traditional Chinese medicine external application composition according to claim 1, characterized in that, The topical Chinese medicine composition also includes one or more of the following auxiliary medicinal materials: 10-20 parts by weight of Sargentodoxa cuneata, 6-14 parts by weight of Clematis chinensis, 6-14 parts by weight of Angelica dahurica, and 2-6 parts by weight of transdermal penetration enhancer, wherein the transdermal penetration enhancer is selected from one or more of camphor, borneol, and musk.

5. The traditional Chinese medicine external application composition according to claim 4, characterized in that, The transdermal penetration enhancer is camphor, which accounts for 1.5-2.5% of the total weight of the topical Chinese medicine composition.

6. The traditional Chinese medicine external application composition according to claim 4, characterized in that, The raw material weight ratio is as follows: The heat-clearing and swelling-reducing group includes 25-35 parts of raw rhubarb and 25-35 parts of charred gardenia. The blood-activating and stasis-removing group includes 10-14 parts of ground beetle, 12-18 parts of vinegar-processed frankincense, 12-18 parts of vinegar-processed myrrh, 8-12 parts of safflower, 10-14 parts of sparganium rhizome, 10-14 parts of turmeric rhizome, and 10-14 parts of vinegar-processed corydalis rhizome. The Qi-regulating and pain-relieving group includes 18-22 parts of Ligusticum chuanxiong, 8-12 parts of Angelica sinensis, 12-18 parts of Sargentodoxa cuneata, 8-12 parts of Clematis chinensis, and 8-12 parts of Angelica dahurica. Transdermal penetration enhancer, including 3-5 parts camphor.

7. The traditional Chinese medicine external application composition according to claim 6, characterized in that, The total weight ratio of the heat-clearing and swelling-reducing group to the blood-activating and stasis-removing group is 1:(1.5~1.8).

8. A method for preparing a medicinal wine from the traditional Chinese medicine external application composition according to claim 6 or 7, characterized in that, Includes the following steps: (1) Weigh each ingredient according to the weight parts, and crush the ingredients except camphor into 10-20 mesh; (2) Mix the raw materials of the heat-clearing and swelling-reducing group and the qi-regulating and pain-relieving group, add 45-60% volume fraction ethanol aqueous solution, the material-liquid ratio is 1: (8-12), and extract at 20-30℃ for 10-20 days; mix the raw materials of the blood-activating and stasis-removing group and extract them separately under the same conditions. (3) Combine the two extracts, filter them with a 180-220 mesh filter cloth, and collect the filtrate; (4) Dissolve camphor in 20-30 times its weight of 95% volume fraction ethanol aqueous solution to prepare camphor alcohol solution, add it at a rate not exceeding 3% of the total volume of filtrate per minute, while stirring at a speed of 100-150 rpm. (5) Let stand for 12-48 hours, then filter.

9. A method for preparing a medicinal powder from the traditional Chinese medicine external application composition as described in any one of claims 1-7, characterized in that, Each of the Chinese medicinal materials is pulverized into a fine powder of 80-120 mesh, mixed evenly according to the formula ratio, and made into a topical powder.

10. The use of a traditional Chinese medicine external application composition as described in any one of claims 1-7 in the preparation of a external medicament for treating soft tissue injuries, characterized in that, When used in the acute phase, the main function is to clear heat and reduce swelling, and it should be used 3-4 times a day; when used in the recovery phase, the main function is to promote blood circulation and remove blood stasis, and it should be used 2-3 times a day.

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