Radix astragali and rhizoma atractylodis healing-promoting oral granules for regulating chronic wound microenvironment as well as preparation process and application of radix astragali and rhizoma atractylodis healing-promoting oral granules

Abstract

The invention belongs to the field of traditional Chinese medicine preparations, and discloses radix astragali and rhizoma atractylodis macrocephalae healing-promoting oral granules for regulating a chronic wound microenvironment, a preparation process of the granules and application of the granules in chronic wound treatment. The radix astragali and rhizoma atractylodis macrocephalae healing promoting oral granules are prepared from 25-35 parts of raw radix astragali, 10-20 parts of radix pseudostellariae, 10-20 parts of fried rhizoma atractylodis macrocephalae, 25-35 parts of radix salviae miltiorrhizae, 10-20 parts of fructus lycii and 10-20 parts of fructus corni through gradient ethanol extraction and astragaloside enrichment processes. According to the granules, systematic regulation of wound surface microenvironment steady state is realized in an oral dosage form for the first time, and triple pathological links of high glucose-infection-ischemia of chronic wound surfaces are synchronously improved through a synergistic effect of a dogwood fruit pulp blood glucose reducing component and a high-proportion astragalus membranaceus / salvia miltiorrhiza healing promoting component; clinical tests show that when the oral granules are singly used for treating the deep chronic refractory ulcer, the healing time is shortened by 32% compared with that of red ointment, and the healing rate is increased to 76.3%.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of traditional Chinese medicine preparations, in particular to a chronic wound microenvironment regulating astragalus and atractylodes granules for oral use, a preparation process thereof and application thereof in the treatment of chronic wounds. BACKGROUND

[0002] The current market of Chinese patent medicines for wound healing is dominated by external preparations, including representative products such as compound cypress liquid, red ointment, and rehabilitation new liquid. These drugs have certain efficacy in clinical application, but have obvious mechanism limitations and application scene restrictions, and are difficult to meet the treatment needs of chronic wounds and other refractory wounds. For example, the treatment plan for diabetic foot ulcers is mainly external preparations (accounting for 90%) of existing Chinese patent medicines, such as compound cypress liquid (antibacterial and anti-inflammatory) and red ointment (promoting granulation and removing necrotic tissue), which have the following limitations: single action: only for local antibacterial or granulation, lacking of systemic regulation of the axis of metabolism-immunity-microcirculation; low healing rate: the healing rate of conventional treatment (debridement + antibiotics) is only 40-60%, and 34% of patients need amputation; dosage form limitation: external ointment / liquid is difficult to penetrate deep ulcers or regulate the pathological environment of the whole body, such as patients with high glucose toxicity and microvascular disease.

[0003] Currently, the research direction of oral traditional Chinese medicines on the market mainly includes two aspects: same component different use and same use different mechanism. In terms of oral traditional Chinese medicines with same component different use, the existing astragalus and atractylodes granules for oral use are mainly used for peptic ulcer, but they are in the form of decoction and are suitable for digestive diseases; oral preparations containing astragalus and salvia miltiorrhiza (such as qi-boosting and blood-activating granules) are mainly used for cardiovascular and cerebrovascular diseases, and are not used for wound healing. In terms of oral traditional Chinese medicines with same use different mechanism, there is a gap in oral wound healing medicines, and there is no similar product for regulating the microenvironment homeostasis of wounds. SUMMARY

[0004] To solve the above problems in the prior art, the present application provides a chronic wound microenvironment regulating astragalus and atractylodes granules for oral use, a preparation process thereof and application thereof in the treatment of chronic wounds, which can effectively regulate the microenvironment homeostasis of chronic wounds by oral administration and is suitable for chronic wound indications requiring local and systemic regulation.

[0005] To achieve the above-mentioned purpose, the present application adopts the following technical solutions:

[0006] The first aspect of the present application is to provide a chronic wound microenvironment regulating astragalus and atractylodes granules for oral use, which comprises the following raw materials by weight: 25-35 parts of raw astragalus, 10-20 parts of prince salvia, 10-20 parts of fried atractylodes, 25-35 parts of salvia miltiorrhiza, 10-20 parts of medlar, and 10-20 parts of wolfberry.

[0007] Preferably, the total proportion of raw astragalus and salvia miltiorrhiza is 50-70%, and the proportion of medlar is ≥12%.

[0008] More preferably, the raw astragalus 30 parts, prince atractylodes 15 parts, fried atractylodes 15 parts, salvia miltiorrhiza 30 parts, medlar 15 parts, and dogwood fruit 15 parts.

[0009] The second aspect of the present application is to provide a preparation process of the Qizhu Cuyu oral granules as described in any of the above, comprising the following steps:

[0010] (1) The prescription amount of raw astragalus and salvia miltiorrhiza is extracted with 70% ethanol gradient for 3 times, and the extract is concentrated to obtain saponin-enriched extract;

[0011] (2) The prescription amount of prince atractylodes and dogwood fruit is extracted with 50% ethanol to obtain loganin and cyclic peptide;

[0012] (3) The prescription amount of fried atractylodes and medlar is water-extracted to obtain polysaccharide;

[0013] (4) The traditional Chinese medicine extract prepared in the above steps (1) to (3) is combined, and after being included with β-cyclodextrin, it is spray dried to prepare Qizhu Cuyu oral granules.

[0014] Preferably, in step (1), the gradient extraction process is dynamic temperature control extraction: 40℃ for 1h, 60℃ for 1.5h, and 80℃ for 2h, and the yield of astragalus saponin is increased by 22.6%.

[0015] The third aspect of the present application is to provide the use of the Qizhu Cuyu oral granules as described above in the preparation of a chronic wound treatment drug, which adopts oral administration.

[0016] Preferably, the chronic wound includes pressure ulcer (bedsore), diabetic foot ulcer, venous ulcer, arterial ulcer, post-traumatic ulcer, or infectious ulcer.

[0017] Preferably, the use of the Qizhu Cuyu oral granules as described above in the preparation of a diabetic foot ulcer treatment drug, which adopts oral administration.

[0018] Preferably, by oral administration, the following is achieved simultaneously: reducing the blood glucose toxicity by regulating the PI3K / Akt pathway, promoting the M2 polarization of macrophages to inhibit inflammation, and increasing the expression of VEGF in the wound to improve the microcirculation.

[0019] Preferably, when treating deep ulcers (depth≥1cm), debridement is combined, and the growth rate of the wound granulation is increased by 40%.

[0020] Preferably, the daily oral dose of the Qizhu Cuyu oral granules is 6-9g, taken 2-3 times.

[0021] The fourth aspect of the present application is to provide a diabetic foot ulcer treatment system comprising the astragalus and aconite oral granules of any one of the above and a wound surface external dressing.

[0022] Preferably, the external dressing is a silver ion antibacterial dressing.

[0023] The fifth aspect of the present application is to provide a method for regulating the microenvironment homeostasis of chronic wounds, comprising orally taking the astragalus and aconite oral granules described above and monitoring the serum IL-6 and VEGF levels.

[0024] The present application adopts the above technical solution, compared with the prior art, has the following technical effects:

[0025] (1) The present application breaks through the limitation of the existing market similar Chinese medicine which can only be used externally, and develops the astragalus and aconite oral granules for chronic wound healing through dosage form innovation. The oral granules can effectively regulate the microenvironment homeostasis of wounds and are suitable for chronic wound indications requiring local and systemic regulation.

[0026] (2) The astragalus and aconite oral granules of the present application use a high proportion of astragalus and salvia miltiorrhiza combination design, and the combination proportion is greater than 60%, which is much higher than the conventional qi and blood circulation formula (such as the commonly used cardiovascular medicine astragalus and salvia miltiorrhiza, which is usually ≤40%). It can effectively improve the core efficacy of the oral granules of tonifying qi and promoting muscle regeneration.

[0027] (3) The present application integrates the blood sugar lowering component of cornus officinalis and the high proportion of astragalus / salvia miltiorrhiza combination for promoting wound healing, which can effectively solve the core contradiction of diabetic foot and provide a systematic solution of internal and external cooperation for the pain point of low healing rate of chronic wounds. The expected effect is significantly better than that of simple external medicine, and the clinical value is high.

[0028] (4) The gradient extraction process of the present application uses dynamic temperature control technology to enrich astragaloside. Compared with the conventional extraction and granulation process, the gradient temperature extraction can effectively solve the problem of thermal sensitive degradation of saponins. The segmented extraction process can avoid the mutual interference of precipitated components when saponins and polysaccharides are co-extracted. And the use of β-cyclodextrin to target the inclusion of each traditional Chinese medicine extract can significantly improve the bioavailability of liposoluble components such as tanshinone, ensuring the wound healing effect. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 The following are the typical treatment effect diagrams of three different treatment schemes in the diabetic foot deep ulcer clinical trial of Example 2, wherein Figure 1 a, 1b and 1c are respectively the foot ulcer wound healing effect diagram of the single control group, the experimental group and the combined group;

[0030] Figure 2 The following is a treatment trend diagram of a case using debridement + astragalus and aconite oral granule scheme in the diabetic foot deep ulcer clinical trial of Example 2. Figure 2a, 2b and 2c are respectively the wound effect pictures of pre-treatment, mid-treatment and post-treatment periods;

[0031] Figure 3 The wound healing effect pictures of two conventional dressing change schemes in the skin deep wound treatment clinical trial of Example 3, wherein Figure 3 a and 3b are respectively the treatment effect pictures of the conventional dressing change scheme and the conventional dressing change + astragalus and zedoary particle oral administration scheme;

[0032] Figure 4 The rat model treatment effect pictures in the experiments of the constant temperature extraction process of Comparative Example 2, wherein Figure 4 a, 4b and 4c are respectively the rat wound healing effect pictures of the gradient extraction group, the constant temperature extraction group and the model control group administration scheme;

[0033] Figure 5 The rat wound healing trend chart of the gradient extraction group administration scheme in the constant temperature extraction process experiment of Comparative Example 2. DETAILED DESCRIPTION

[0034] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments.

[0035] Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without making creative efforts are within the scope of protection of the present application.

[0036] At present, there are many similar Chinese patent medicines on the market, but there is no oral medicine for treating chronic wound healing, and most of them are drugs with the effect of promoting blood circulation to remove blood stasis, which are used to repair wound healing by the concept, and the existing products such as compound cortex phellodendri liquid and red ointment mainly focus on single mechanism (such as simple antibacterial or promoting granulation growth), lack of systematic quantitative regulation of wound microenvironment homeostasis; or some are hospital experience drugs or self-made preparations. And the clinical research shows that: the healing rate of conventional compound cortex phellodendri liquid, red ointment and similar Chinese patent medicines for treating diabetic foot ulcers is only 40-60%, and 90% of these wound Chinese medicines are external ointments, and there is no similar product for chronic wounds at present.

[0037] To solve the above problems, fill the market gap, based on the inventor's deep participation and published research results "Exploration of TCM diagnosis and treatment of refractory ulcer based on local qi theory" (published in Shanghai Journal of Chinese Medicine, manuscript number z20250505003, Chen Lili, Cao Yongzhi, Li Feng, Lu Jinggen), combined with clinical research and application requirements in actual work, the first oral granules are created in the field of wound healing, and a Qizhu Cuyu oral granule for chronic wounds is developed. The core scheme of this oral dosage form is to achieve the purpose of breaking through the bottleneck of low healing rate of chronic wounds such as diabetic foot ulcer by reducing blood sugar (Shanyu Rou) and promoting healing (high proportion of Huangqi / Danshen) synergistic effect, directly attacking the pathological link of the whole body-local, which has significant clinical application value.

[0038] Specifically, the present application provides a Qizhu Cuyu oral granule for regulating wound microenvironment homeostasis by oral administration, comprising the following raw materials by weight: 25-35 parts of raw Huangqi, 10-20 parts of Taizishen, 10-20 parts of Fried Baizhu, 25-35 parts of Danshen, 10-20 parts of Gouqi, and 10-20 parts of Shanyu Rou.

[0039] As a preferred scheme, in the formula of the Qizhu Cuyu oral granule, the total proportion of raw Huangqi and Danshen is 50-70%, and the proportion of Shanyu Rou is ≥12%.

[0040] As one of the optimal embodiments, the prescription of the Qizhu Cuyu oral granule is: 30 parts of raw Huangqi, 15 parts of Taizishen, 15 parts of Fried Baizhu, 30 parts of Danshen, 15 parts of Gouqi, and 15 parts of Shanyu Rou.

[0041] The traditional effects of the above components and their mechanisms of action in the oral granule of the present application are shown in Table 1 as follows:

[0042] Table 1 Comparison of the mechanisms of action and traditional effects of the formula of the oral granule of the present application:

[0043] Composition Composition Functional positioning Synergistic mechanism Traditional efficacy Radix Astragali 25～35 Benefiting Qi and Securing Exterior Immune regulation + promoting angiogenesis (main drug) Supplementing Qi and Promoting Granulation Radix Pseudostellariae 10～20 Benefiting Qi and Nourishing Yin Improving glucose metabolism + cell activity Benefiting Qi and Nourishing Yin Rhizoma Atractylodis Macrocephalae 10～20 Intestinal barrier repair Enhancing intestinal immunity + regulating systemic inflammation Invigorating Spleen and Drying Dampness Radix Salviae Miltiorrhizae 25～35 Activating Blood and Removing Stasis Antithrombotic + increasing blood perfusion Activating Blood and Removing Stasis Fructus Lycii 10～20 Nourishing Liver and Kidney Antioxidation + anti-apoptosis Nourishing Liver and Kidney Fructus Corni 10～20 Nourishing Liver and Kidney Lowering blood glucose + regulating immunity Nourishing Liver and Kidney

[0044] The Qizhu Cuyu oral granule for chronic wound treatment provided by the present application, taking diabetic foot ulcer as an example, has the following main technical solutions: first, a high proportion of Huangqi + Danshen combination scheme (about 60%) is adopted, which far exceeds the conventional Qi-tonifying and Blood-activating formula (such as Huangqi Danshen for cardiovascular drugs, which usually accounts for ≤40%), highlighting the core of Qi-tonifying and Muscle-regenerating; second, a combination scheme of Shanyu Rou (hypoglycemic) and Taizishen (improving insulin resistance) suitable for chronic wounds such as diabetes is adopted to directly attack the pathological basis of high glucose toxicity of diabetic foot; third, the oral granule of the present application is different from similar formulas, and the traditional Qizhu Cuyu decoction for peptic ulcer does not contain Shanyu Rou / Gouqi, and is a decoction rather than a granule, while the traditional weight loss and lipid-lowering formula (such as published patent CN120285097A) contains Shanyu Rou / Gouqi, but does not contain Huangqi / Danshen and has a different purpose.

[0045] The present application breaks through the limitation of no clear indication or only external use of the same kind of existing marketed Chinese medicine, and develops the oral granules for wound healing through dosage form innovation, which can effectively regulate the microenvironment homeostasis of wound, is suitable for the indication of diabetic foot ulcer requiring systemic regulation and local regulation, and breaks through the limitation of existing single mechanism. On the one hand, oral administration of the granules can reduce blood sugar (astragalus root and radix pseudoginseng), reduce the inhibition of glycosylated end products on fibroblasts, improve circulation (salvia miltiorrhiza and astragalus root), increase blood perfusion of ischemic tissues in lower limbs, and achieve systemic regulation; on the other hand, oral administration of the granules can promote the growth of granulation tissue (astragalus saponin), angiogenesis (tanshinone), and epithelialization (radix pseudoginseng cyclic peptide), synchronously solve the triple obstacles of high glucose-infection-ischemia, and increase the healing rate from 40-60% to more than 70%, thereby achieving the purpose of local regulation.

[0046] To avoid the mutual interference of each prescription in the process of preparing traditional Chinese medicine granules by extraction and granulation process, fully exert the efficacy of each Chinese medicine prescription, and improve the utilization of active ingredients. The present application provides a preparation process of Qizhu Cuyu oral granules, which comprises the following steps: (1) extracting prescription amount of raw astragalus and salvia miltiorrhiza with 70% ethanol for 3 times, and concentrating the combined extract to obtain saponin-enriched extract; (2) extracting prescription amount of radix pseudoginseng and astragalus root with 50% ethanol to obtain loganin and cyclic peptide; (3) water-extracting prescription amount of fried atractylodes and medlar to obtain polysaccharide; and (4) combining the Chinese medicine extracts prepared in steps (1) to (3), adding β-cyclodextrin after inclusion, and then spray drying and granulating to obtain Qizhu Cuyu oral granules.

[0047] Preferably, in step (1), the process of extracting prescription amount of radix pseudoginseng and astragalus root with 50% ethanol to obtain loganin and cyclic peptide specifically comprises the following steps:

[0048] (11) Raw material treatment: 30 kg of raw astragalus and 30 kg of salvia miltiorrhiza are pulverized through a 40-mesh sieve for standby;

[0049] (12) Gradient extraction:

[0050] First extraction: the pulverized raw astragalus and salvia miltiorrhiza are added with 10 times the amount of 70% ethanol, and dynamic stirring is performed at 40°C for 1 h (rotation speed 120 rpm);

[0051] Second extraction: the filter residue after the first extraction is added with 8 times the amount of 70% ethanol, and extraction is performed at 60°C for 1.5 h;

[0052] Third extraction: the filter residue after the second extraction is added with 6 times the amount of 70% ethanol, and extraction is performed at 80°C for 2 h.

[0053] (13) Concentration and purification: combine the three extracts of step (12), concentrate at 60℃ under reduced pressure to a relative density of 1.15 (60℃), then add 2% of the volume of the concentrated liquid ZTC clarifier to remove impurities, stand for 24h, then take the supernatant, and treat by ultrafiltration (10kDa membrane) to obtain a saponin-rich extract (astragaloside content ≥4.2mg / g).

[0054] Preferably, in step (2), the process of extracting loganin and cyclic peptides from the prescribed amount of Radix Pseudostellariae and Fructus Corni with 50% ethanol, specifically includes the following steps:

[0055] (21) Mixed extraction: take 15kg of Radix Pseudostellariae and 15kg of Fructus Corni, crush them through a 30-mesh sieve, then add 12 times the amount of 50% ethanol, and reflux extract at 80℃ for 2 times, 1.5h each time;

[0056] (22) Concentration: combine the extracts of step (21), concentrate at 55℃ under reduced pressure to remove the alcohol taste, and stop the concentration when the loganin content is ≥1.8mg / g (HPLC method), to obtain a loganin and cyclic peptide-rich extract.

[0057] Preferably, in step (3), the process of water-extracting polysaccharides from the prescribed amount of Fried Atractylodes Rhizome and Fructus Lycii, specifically includes the following steps:

[0058] (31) Water extraction process: add 10 times the amount of water to 15kg of Fried Atractylodes Rhizome and 15kg of Fructus Lycii, and micro-boil decoct for 2 times (2h for the first time and 1.5h for the second time), with pH controlled at 6.5-7.0;

[0059] (32) Refining: combine the decoctions of step (31), centrifuge to remove the residue (4000rpm, 15min), then add 3% chitosan flocculation to the supernatant, stand for 12h, then filter, and concentrate to a polysaccharide content of ≥25% (determined by the phenol-sulfuric acid method) to obtain a polysaccharide-rich extract.

[0060] Preferably, in step (4), the process of combining the traditional Chinese medicine extracts obtained in steps (1) to (3) above, adding β-cyclodextrin inclusion, and then spray drying and granulating, specifically includes the following steps:

[0061] (41) Mixed inclusion: combine the traditional Chinese medicine extracts obtained in steps (1), (2) and (3) (temperature ≤50℃), add 30% β-cyclodextrin (i.e. drug solution: carrier = 1:0.3) to the mixed extract, and stir at 45℃ for 1h (rotation speed 80rpm) to include;

[0062] (42) Spray drying: control the inlet air temperature at 180±5℃, the outlet air temperature at 90±2℃, the atomization pressure at 0.25MPa, and the feeding speed at 20mL / min to spray granulate, and the obtained granules have a water content of ≤5% (Karl Fischer method).

[0063] (43) The granules prepared in step (42) are sieved through an 18-mesh sieve to size, and are packed in nitrogen-filled aluminum plastic bags, with each bag containing 2 g (equivalent to 12 g of crude drug).

[0064] In the above preparation process, the medicinal extractives of each group of prescriptions are first extracted by gradient temperature control, and the extractives of Huangqi and Danshen are extracted at 40°C for 1 h to start the protection activity and solve the problem of saponin thermal degradation; meanwhile, each group of prescriptions is divided into three groups for extraction, and a segmented extraction strategy is adopted to avoid mutual interference of components, such as precipitation produced when saponin and polysaccharide are co-extracted; finally, β-cyclodextrin is used for targeted inclusion in the process of spray granulation, which can significantly improve the bioavailability of fat-soluble components (such as tanshinone).

[0065] In addition, the application also provides a use of the Qizhu Cuyu oral granules in preparation of a medicament for treating chronic wounds, which can simultaneously achieve the following effects by oral administration: the active ingredient loganin can regulate the PI3K / Akt pathway to reduce the toxicity of hyperglycemia, the active ingredient astragalus polysaccharide can promote the M2 polarization of macrophages to inhibit inflammation, and the active ingredient tanshinone can significantly increase the expression of VEGF on the wound surface to improve the microcirculation, thereby increasing the healing rate of diabetic foot ulcers from 40-60% to more than 70%.

[0066] Preferably, when treating deep ulcers (depth≥1 cm), the Qizhu Cuyu oral granules are combined with debridement, and the growth rate of wound granulation can be increased by more than 40%. The daily oral dose of the Qizhu Cuyu oral granules is 6-9 g, which is taken 2-3 times a day.

[0067] The following examples and comparative examples verify the creative breakthroughs of the application in dosage form innovation, prescription synergy and process optimization through specific prescriptions, processes, test data and conclusion analysis.

[0068] Example 1: Preparation of base granules

[0069] A Qizhu Cuyu oral granule for regulating the microenvironment of chronic wounds, which has the following prescription: 30 kg of raw Huangqi, 15 kg of Taizishen, 15 kg of fried Baishu, 30 kg of Danshen, 15 kg of Gouqi, and 15 kg of Shanyurou.

[0070] Based on the above prescription, the preparation process of the Qizhu Cuyu oral granule of this example includes the following steps:

[0071] (1) Gradient extraction of Huangqi and Danshen:

[0072] The prescription amount of medicinal materials Huangqi and Danshen are crushed through a 40-mesh sieve and reserved;

[0073] After mixing the powder, perform segmented extraction, first segment: dynamic extraction at 40°C (70% ethanol, 10 times the amount, 120 rpm, 1 h), second segment: extraction at 60°C (70% ethanol, 8 times the amount, 1.5 h), third segment: extraction at 80°C (70% ethanol, 6 times the amount, 2 h);

[0074] After combining the extracts, concentrate at 60°C under reduced pressure to a density of 1.15, then add 2% ZTC clarifying agent to remove impurities, and finally treat by ultrafiltration (10 kDa membrane) to obtain a saponin-enriched extract with an astragaloside A content of ≥4.2 mg / g.

[0075] (2) Extraction of Radix Pseudostellariae and Fructus Corni:

[0076] Grind the prescription amount of Radix Pseudostellariae and Fructus Corni to pass through a 30-mesh sieve, then add 12 times the amount of 50% ethanol, and reflux extract at 80°C for 2 times, each for 1.5 h;

[0077] Combine the above extracts, concentrate at 55°C under reduced pressure to remove the alcohol taste, and stop concentrating when the loganin content is ≥1.8 mg / g (HPLC method) to prepare a loganin and cyclic peptide-enriched extract.

[0078] (3) Extraction of Fried Rhizoma Atractylodis Macrocephalae and Fructus Lycii:

[0079] Add 10 times the amount of water to the prescription amount of Fried Rhizoma Atractylodis Macrocephalae and Fructus Lycii, and boil decoct for 2 times (2 h for the first time and 1.5 h for the second time), with pH controlled at 6.8;

[0080] Combine the above decoctions, centrifuge to remove the residue (4000 rpm, 15 min), then add 3% chitosan flocculation to the supernatant, stand for 12 h, filter, then concentrate to a polysaccharide content of ≥25% (determined by the phenol-sulfuric acid method) to prepare a polysaccharide-enriched extract.

[0081] (4) Integrated granulation:

[0082] Combine the traditional Chinese medicine extracts obtained in steps (1), (2), and (3) (temperature ≤50°C), add β-cyclodextrin accounting for 30% of the weight of the mixed extract (i.e. drug solution: carrier = 1:0.3) to the mixed extract, and stir to package at 45°C for 1 h (rotation speed 80 rpm);

[0083] Control the inlet air temperature at 180±5°C, the outlet air temperature at 90±2°C, the atomization pressure at 0.25 MPa, and the feeding speed at 20 mL / min to perform spray granulation, and the prepared granules have a water content of ≤5% (Karl Fischer method);

[0084] Pass the prepared granules through an 18-mesh sieve to perform whole-grain processing, use nitrogen-filled aluminum plastic packaging, and control each bag to contain 2 g (equivalent to 12 g of crude drug).

[0085] The prepared Jiuzhu Cuyu oral granules of this embodiment are respectively subjected to active ingredient determination and solubility test, and the test data are shown in Table 2.

[0086] Table 2 test results

[0087] Index Test value Standard of Chinese Pharmacopoeia Content of Astragaloside IV 4.21 ± 0.15 mg / g ≥ 3.5 mg / g Content of Tanshinone IIA 2.38 ± 0.11 mg / g ≥ 1.8 mg / g Content of Loganin 1.85 ± 0.07 mg / g ≥ 1.2 mg / g Granule dispersion time 3.2 ± 0.3 min ≤ 10 min

[0088] Conclusion analysis: According to the active ingredient determination results in Table 1, the gradient extraction process adopted in the application can increase the retention rate of heat-sensitive saponins (astragaloside A) to 96.4% (73.8% for the conventional process), and the solubility of tanshinone reaches 92.3% (68.7% for the uncoated group) through β-cyclodextrin inclusion, which meets the needs of industrial production.

[0089] Example 2: Clinical trial of diabetic foot deep ulcer

[0090] Test design: 186 subjects of Wagner 2-3 grade diabetic foot ulcer patients (wound depth 1.2-3.5 cm) are divided into three groups, and different treatment schemes are used for clinical research.

[0091] Specific grouping is as follows: control group: n = 62, using debridement + red oil paste external treatment scheme; experimental group: n = 62, using debridement + Qizhu granules oral (8g / day) scheme; combined group: n = 62, using debridement + Qizhu granules (8g / day) + red oil paste scheme.

[0092] Evaluation index: healing time, 12-week healing rate, granulation growth rate, histological analysis. The specific test data and mechanism verification data are shown in Tables 3 and 4.

[0093] Table 3 test results

[0094] Index Control group Experimental group Combined group Average healing time (days) 41.2±5.7 28.1±4.3 24.3±3.9 12-week healing rate 52.7％ 76.3％ 84.1％ Granulation growth rate (mm / d) 0.38±0.06 0.53±0.05 0.61±0.07

[0095] Table 4 histological mechanism verification (treatment for 4 weeks, n = 20):

[0096] Index Control group Experimental group Improvement rate Number of new capillaries (pieces / HP) 12.7±3.2 24.3±4.1 +91.3％ Collagen fiber density (%) 38.5±6.7 61.2±7.9 +58.9％ TNF-α positive cell rate (%) 29.8±5.1 11.3±2.7 -62.1％

[0097] Conclusion analysis: Compared with the traditional external medicine, the simple oral granules can shorten the healing time by 32% (p < 0.001), which indicates that the loganin in the oral granules can significantly reduce the fasting blood glucose (6.8 ± 0.7 vs 8.9 ± 1.1 mmol / L, p < 0.01), the active ingredient tanshinone can increase the expression of VEGF in the wound by 2.1 times (312 ± 45 vs 149 ± 32 pg / mL), and the high proportion of astragalus (30%) design scheme can promote the polarization of macrophage M2 (CD206+ cells increase by 67.4%).

[0098] In addition, asFigure 1 The results of the three different treatment options are shown in the following figures, in which Figure 1 a, 1b and 1c are respectively the healing effect pictures of a single lower limb tibial anterior chronic vascular ulcer, and it is obvious that the healing effect of the debridement + Qizhu granules oral treatment scheme is more significant than that of the traditional dressing method treatment. And Figure 2 Further shows the treatment trend chart of one of the diabetic foot refractory ulcer patients using debridement + Qizhu granules oral scheme, in which Figure 2 a, 2b and 2c are respectively the wound effect pictures at the early, middle and late stages of treatment.

[0099] Example 3: Clinical trial of deep skin wound treatment

[0100] Test design: 60 cases of refractory ulcer patients (wound / fistula depth ≥4cm) were divided into control group and treatment group, in which: control group (n=30), using conventional dressing scheme; treatment group (n=30), using conventional dressing + Qizhu granules oral (8g / day) scheme.

[0101] Evaluation index: wound / fistula closure time, deep granulation coverage rate, 6-month recurrence rate.

[0102] The test data and drug permeability detection results after anal fistula surgery are shown in Table 5 and Table 6 as follows.

[0103] Table 5 Test results:

[0104] Index Control group Treatment group Wound / sinus closure time (days) 35.6±6.2 22.4±4.8 Deep granulation coverage rate 47.3％±8.1％ 82.6％±7.5％ 6-month recurrence rate 23.3％ 6.7％

[0105] Table 6 Drug permeability detection (LC-MS / MS method):

[0106]

[0107]

[0108] Conclusion analysis: From the test results of Table 5 and Table 6 above, it can be seen that oral administration of Qizhu granules can penetrate into deep wounds / fistula (4cm drug concentration >1μg / g), repair intestinal barrier through the active factor of stir-baked atractylodes (serum DAO activity decreased by 42.7%), inhibit deep oxidative stress (MDA decreased by 58.2%) through polysaccharide of medlar, and increase the granulation coverage rate by 74.6% (p<0.001), which proves that the oral granules are also effective for non-diabetic deep wounds.

[0109] Figure 3 The wound healing effect pictures of a group of sacrococcygeal decubitus patients using conventional dressing scheme and conventional dressing + Qizhu granules oral scheme respectively are shown, in which Figure 3 a is the treatment effect picture using conventional dressing scheme,Figure 3 b is the treatment effect diagram of the conventional dressing change + the oral regimen of the invention of the Astragalus and Atractylodes granules.

[0110] Comparative Example 1: Cortex Acanthopanacis Senticosi Deletion Experiment

[0111] An Astragalus and Atractylodes Promoting Healing Oral Granule, with Cortex Acanthopanacis Senticosi deleted, and the rest the same as Example 1. Its prescription is: Radix Astragali 30 kg, Radix Notoginseng 15 kg, Atractylodes 15 kg, Salvia miltiorrhiza 30 kg, Fructus Lycii 15 kg.

[0112] The preparation process of the Astragalus and Atractylodes Promoting Healing Oral Granule is different from Example 1 in that in step (2), only Radix Notoginseng is extracted with 50% ethanol by reflux extraction twice, and the rest of the process remains unchanged.

[0113] The Astragalus and Atractylodes Promoting Healing Oral Granule prepared in Comparative Example 1 was subjected to clinical trials according to the verification method of Example 2: 40 cases of Wagner 2-3 grade diabetic foot ulcer patients (wound depth 1.2-3.5 cm) were divided into complete prescription group and Cortex Acanthopanacis Senticosi deletion group, and different treatment regimens were used for clinical research.

[0114] Evaluation index: fasting blood glucose, healing time, 12-week healing rate, EGF serum level and IL-6 reduction amplitude. The specific test data are shown in Table 7.

[0115] Table 7 Diabetic foot patient test (n=40):

[0116] Index Complete formula group Fructus Corni deletion group Fasting blood glucose (mmol / L) 6.8±0.7 8.1±0.9 Wound healing time (days) 28.1±4.3 37.2±5.6 Serum VEGF level (pg / mL) 312±45 218±38 IL-6 reduction rate 62.3％ 38.7％

[0117] Conclusion analysis: From the test data in Table 7, the absence of Cortex Acanthopanacis Senticosi in the oral granule of Comparative Example 1 leads to insufficient activation of the PI3K / Akt pathway (p-Akt expression decreased by 54.2%), and the continuous inhibition of fibroblast migration in a high glucose environment (wound healing rate in scratch test: 68.4% vs 42.3%). Therefore, this test proves that the compatibility of Cortex Acanthopanacis Senticosi in the prescription has the dual effects of hypoglycemia (loganin) and immunomodulation (ursolic acid), which can shorten the wound healing time by 24.4% (p<0.001), and its absence can easily lead to uncontrolled blood glucose (>7.8 mmol / L) and inhibition of VEGF expression, confirming its irreplaceability in the prescription.

[0118] Comparative Example 2: Constant temperature extraction process experiment

[0119] An Astragalus and Atractylodes Promoting Healing Oral Granule, its prescription is: Radix Astragali 30 kg, Radix Notoginseng 15 kg, Atractylodes 15 kg, Salvia miltiorrhiza 30 kg, Fructus Lycii 15 kg, Cortex Acanthopanacis Senticosi 15 kg.

[0120] Based on the above prescription, the preparation process of Comparative Example 2 is different from the example in that step (1) adopts constant temperature extraction process in the extraction process of Huangqi and Danshen, and only constant temperature extraction at 80°C for 4.5h (the total time is the same as that of Example 1), and the rest of the steps are consistent with the preparation process of Example 1.

[0121] The Qizhu Cuyu oral granules prepared by the gradient extraction process of Example 1 are the gradient extraction group, and the Qizhu Cuyu oral granules prepared by the constant temperature extraction process of Comparative Example 2 are the constant temperature extraction group. The ingredients of the two groups are tested, and the test data are shown in Table 8.

[0122] Table 8 Ingredient data of granules prepared by different extraction processes

[0123] Index Gradient extraction group Constant temperature extraction group Reduction rate Retention rate of Astragaloside IV 96.4％±2.1％ 73.8％±3.5％ -22.6％ Retention rate of Tanshinone IIA 94.7％±1.8％ 70.2％±4.1％ -24.5％ Total saponin yield 8.72％ 6.15％* -29.5％

[0124] In addition, the Qizhu Cuyu oral granules prepared by Example 1 and Comparative Example 2 are respectively subjected to rat wound model test, and the specific test method is as follows:

[0125] 1. Establishment of animal model

[0126] Animal strain: SPF level SD rats, male, body weight 200±20g (purchased from Sibeifu Biological)

[0127] Diabetes induction: intraperitoneal injection of streptozotocin (STZ, 65mg / kg, dissolved in 0.1M citric acid buffer, pH4.5), and rats with fasting blood glucose≥16.7mmol / L after 72h were selected;

[0128] Wound making: After the selected rats were anesthetized (3% sodium pentobarbital, 40mg / kg), the back was shaved, and the full-thickness skin was removed (diameter 1.8cm circular wound, deep to the fascia layer) under sterile operation, so that the wound interval was≥3cm to prevent fusion.

[0129] 2. Grouping and administration, as shown in Table 9:

[0130] Group Treatment plan (n = 10) Dose Gradient extraction group Qizhu granules (gradient process) 1.0 g / kg / d (gavage) Constant temperature extraction group Qizhu granules (80°C constant temperature process) 1.0 g / kg / d (gavage) Model control group Same volume of normal saline -

[0131] Course of treatment: continuous administration for 14 days;

[0132] Granule preparation: before use, 0.2g / mL suspension was prepared with distilled water.

[0133] 3. Test results

[0134] Table 10 Comparison of wound healing time

[0135] Group 50% healing time (days) Complete epithelialization time (days) Gradient extraction group 6.8±0.9 15.3±1.2 Constant temperature extraction group 9.1±1.3 20.7±1.8 Model control group 11.5±1.7 26.4±2.1

[0136] Table 11 Histological quantitative results (14th day)

[0137] Index Gradient extraction group Constant temperature extraction group Reduction rate Number of CD31+ blood vessels (pieces / HP) 28.4±3.1 18.9±2.7 -33.5％ Collagen area ratio (%) 42.6±5.3 28.7±4.1 -32.6％ VEGF concentration (pg / mg protein) 38.5±4.2 24.1±3.3 -37.4％

[0138] Conclusion analysis: From the test results of the above Table 10 and Table 11, it can be seen that the constant temperature extraction process of Comparative Example 2 leads to a large amount of degradation (loss of nearly 30%) of heat-sensitive saponins, a 35.3% delay in wound healing (p<0.001), and a 33.5% reduction in capillary density (p<0.01), which directly weakens the pro-angiogenic effect, thereby confirming the key value of the gradient extraction process of the present application for the protection of active ingredients.

[0139] In addition, Figure 4 The rat model treatment effect diagram using the gradient extraction group, the constant temperature extraction group and the model control group dosing regimen respectively is shown, wherein 4a is the rat wound healing effect diagram of Qizhu Cuyu oral granules prepared by the gradient extraction process of Example 1, 4b is the rat wound healing effect diagram of Qizhu Cuyu oral granules prepared by the constant temperature extraction process of Comparative Example 2, Figure 4 c is the rat wound healing effect diagram using the same volume of normal saline regimen, and it is obvious that the rat wound healing effect of the gradient extraction group of the present application is the most excellent. Figure 5 The rat wound healing trend diagram using the gradient extraction group dosing regimen is shown.

[0140] Comparative Example 3: dosage form replacement test

[0141] A Qizhu Cuyu external use gel has the prescription of: 30 kg of raw Astragalus membranaceus, 15 kg of Prince Ginseng, 15 kg of fried Atractylodes, 30 kg of Salvia miltiorrhiza, 15 kg of Fructus Lycii, and 15 kg of Fructus Corni. Different from Example 1, the traditional technology is used to prepare a carbomer gel (containing 10% of the drug) from the traditional Chinese medicine extract prepared in steps (1) to (3).

[0142] The Qizhu Cuyu oral granules prepared in the above Example 1 are used as the oral granule group, and the carbomer gel (containing 10% of the drug) prepared by the same prescription of Comparative Example 3 is used as the external use gel group, and the pig skin deep ulcer model test is carried out, and the specific test method is as follows:

[0143] 1. Model establishment and grouping

[0144] Animals: Bama miniature pigs, male, body weight 35±3 kg (barrier environment feeding);

[0145] Deep ulcer preparation: After anesthesia (50, 5 mg / kg intramuscular injection), the back was shaved and disinfected; trepanning of skin: 2 cm diameter circular full-thickness skin (deep to subcutaneous fat layer, about 1.5 cm); deep extension: using a sterile spatula to peel down to the subfascial layer to form a conical wound cavity with a depth of 2.5 cm (simulating clinical deep ulcer), and the wound interval was ≥5 cm to prevent cross infection.

[0146] Grouping design (n=6 wounds / group):

[0147] Group Dosing regimen Dose / frequency Oral granule group Qizhu Shuyu granules suspension gavage 0.5 g / kg / d (equivalent to human equivalent dose) Topical gel group 10% Qizhu extract-containing carbomer gel applied to the wound Daily dressing change, thickness 2 mm Blank control group Normal saline gavage + blank gel application -

[0148] 2. Drug administration and sample collection

[0149] (1) Drug administration regimen

[0150] Oral group: Granules were prepared into a 0.1 g / mL suspension with distilled water and administered orally in the morning

[0151] External use group: The gel was evenly applied to the wound surface and fixed with sterile gauze

[0152] Course of treatment: 14 consecutive days.

[0153] 3. Test results

[0154] (1) Drug penetration concentration in deep tissues (7th day)

[0155]

[0156]

[0157] (2) Comparison of wound healing effects

[0158] Index Oral group Topical group Blank group Granulation growth rate (mm 3 / d) 152±18 64±12 42±10 Wound cavity closure rate (14th day) 78.3％±6.7％ 35.2％±5.1％ 22.6％±4.3％ Deep necrotic tissue residue 9.2％±2.1％ 41.7％±7.3％ 53.8％±8.5％

[0159] (3) Systemic regulation

[0160] Index Oral group (change rate) Topical group (change rate) Serum IL-6 ↓62.3％ ↓8.7％ Fasting blood glucose ↓ 2.4 mmol / L + 0.3 mmol / L Periwound blood flow ↑34.7％ ↑5.2％

[0161] 4. Histological evidence

[0162] H&E staining of deep tissues (2.5 cm) (14th day):

[0163] Group Granulation tissue characteristics Inflammatory state Oral group Dense new capillaries (↑↑) Few lymphocyte infiltrates Topical group Sparse blood vessels, large amount of necrotic debris (↓↓) Intensive neutrophil infiltration (+++) Blank group No vascular neogenesis, extensive necrosis (↓↓↓) Purulent inflammation (++++)

[0164] Conclusion analysis: From the above test results of the pig skin deep ulcer model, it can be seen that the oral granules of the present application can deliver the effective component to the deep tissues of 2.5 cm (tanshinone > 1.97 μg / g, 5 times the minimum inhibitory concentration), and the specific significant deep penetration; in terms of systemic-local coordination, it can reduce systemic blood glucose (-2.4 mmol / L) and IL-6 (62.3% reduction), and promote the growth of local deep granulation (137.5% increase in growth rate compared with the external use group). The effective component of the same group of external use gel was not detected at a depth of 1.5 cm (concentration <0.05 μg / g), and it could not improve the systemic pathological environment (blood glucose and inflammation did not change), which directly proves that the oral granules proposed in the present application are an effective solution to the deep ulcer drug delivery bottleneck, highlighting the irreplaceability of the oral granules in the treatment of deep wounds.

[0165] The foregoing detailed description of the application has been presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the application to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The described embodiments were chosen in order to best explain the principles of the application and its practical application to thereby enable others skilled in the art to best utilize the application. It is intended that the described embodiments be considered merely exemplary and that all modifications and variations are possible given the scope of the application.

Claims

1. An oral granule for regulating the microenvironment of chronic wounds and promoting healing, characterized in that, It contains the following ingredients by weight: 25-35 parts raw Astragalus membranaceus, 10-20 parts Codonopsis pilosula, 10-20 parts stir-fried Atractylodes macrocephala, 25-35 parts Salvia miltiorrhiza, 10-20 parts Lycium barbarum, and 10-20 parts Cornus officinalis.

2. The Astragalus and Atractylodes Macrocephala Oral Granules for Regulating the Microenvironment of Chronic Wounds according to claim 1, characterized in that, The total proportion of raw Astragalus membranaceus and Salvia miltiorrhiza is 50-70%, and the proportion of Cornus officinalis is ≥12%.

3. The Astragalus and Atractylodes Macrocephala Oral Granules for Regulating the Microenvironment of Chronic Wounds according to claim 1 or 2, characterized in that, It contains the following ingredients by weight: 30 parts raw Astragalus membranaceus, 15 parts Codonopsis pilosula, 15 parts stir-fried Atractylodes macrocephala, 30 parts Salvia miltiorrhiza, 15 parts Lycium barbarum, and 15 parts Cornus officinalis.

4. A preparation process for the Astragalus and Atractylodes Promoting Healing Oral Granules as described in any one of claims 1 to 3, characterized in that, Includes the following steps: (1) Extract the prescribed amount of raw Astragalus membranaceus and Salvia miltiorrhiza three times with 70% ethanol, combine the extracts and concentrate to obtain saponin-enriched extract. (2) Extract loganin and cyclic peptides from the prescribed amount of Codonopsis pilosula and Cornus officinalis using 50% ethanol; (3) Extract polysaccharides from the prescribed amount of stir-fried Atractylodes macrocephala and Lycium barbarum; (4) Combine the Chinese herbal extracts obtained in steps (1) to (3) above, add β-cyclodextrin for encapsulation, and then granulate by spray drying to obtain Astragalus and Atractylodes Promoting Healing Oral Granules.

5. The preparation process according to claim 4, characterized in that, In step (1), the gradient extraction process is a dynamic temperature-controlled extraction: extraction at 40℃ for 1 hour, at 60℃ for 1.5 hours, and at 80℃ for 2 hours.

6. The use of the Astragalus and Astragalus Promoting Healing Oral Granules as described in any one of claims 1 to 3 in the preparation of a drug for treating chronic wounds, wherein the granules are administered orally.

7. The application according to claim 6, characterized in that, The daily oral dosage of the Astragalus and Atractylodes Promoting Healing Oral Granules is 6-9g, divided into 2-3 doses.

8. The application according to claim 6, characterized in that, The chronic wounds include pressure ulcers (bedsores), diabetic foot ulcers, venous ulcers, arterial ulcers, post-traumatic ulcers, or infected ulcers.

9. A chronic wound treatment system, characterized in that, It includes the Astragalus and Atractylodes Promoting Healing Oral Granules and Wound Dressings as described in any one of claims 1 to 3.

10. A method for regulating the homeostasis of the microenvironment in chronic wounds, characterized in that, Including oral administration of the Astragalus and Astragalus Promoting Healing Oral Granules as described in any one of claims 1 to 3 and monitoring serum IL-6 and VEGF levels.

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