External traditional Chinese medicine composition for promoting healing of chronic refractory wounds as well as preparation method and application of external traditional Chinese medicine composition

Abstract

The invention provides an external traditional Chinese medicine composition for promoting healing of chronic refractory wounds as well as a preparation method and application of the external traditional Chinese medicine composition. The composition is prepared from water-soluble lignin extracted from callicarpa nudiflora stems and callicarpa nudiflora leaf water extract in a weight ratio of (1-3): (1-3). The preparation method comprises the following steps: extracting water-soluble lignin from callicarpa nudiflora stems; effective components are extracted from the callicarpa nudiflora leaves through water decoction; and finally, mixing the two with pharmaceutical auxiliary materials to prepare external preparations such as gel and the like. The composition disclosed by the invention gives full play to the anti-oxidation and repair promotion capabilities of the water-soluble lignin and the traditional effects of clearing away heat and toxic materials and astringing to stop bleeding of the callicarpa nudiflora, and has a multi-target synergistic effect. Animal experiments show that the composition can obviously accelerate the healing of chronic refractory wounds, the curative effect is definite, the preparation process is stable, the safety is high, and a new effective medicine is provided for solving the problem of chronic wound healing.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and in particular to a topical traditional Chinese medicine composition for promoting the healing of chronic and refractory wounds, its preparation method, and its application. Background Technology

[0002] Chronic, non-healing wounds refer to wounds that cannot achieve anatomical and functional integrity through a normal, orderly, and timely repair process. They typically refer to wounds that have not healed or show no tendency to heal after more than one month. These wounds are commonly seen in diabetic foot ulcers, pressure ulcers, venous ulcers, and wound infections. Their pathological characteristics include recurrent local infections, microcirculatory disturbances, tissue edema, growth factor deficiency, and abnormal immune regulation. In my country, patients hospitalized due to chronic wounds account for approximately 1.7% of all hospitalized patients. With an aging population, non-healing wounds in my country are mainly caused by diabetic foot ulcers and complications of chronic diseases in the elderly (such as wounds caused by arterial diseases and pressure ulcers). These wounds are not only deeper but also often more complex and variable, becoming a significant factor affecting the health of the elderly. Although chronic, non-healing wounds do not immediately threaten life, their prolonged non-healing over months, years, or even decades severely impacts the patient's recovery from the primary disease and quality of life, causing significant psychological stress and placing a heavy burden of care and financial strain on families.

[0003] The mechanisms of chronic wound repair are not fully understood, but they are characterized by recurrent bacterial infections, persistent deficiency, abnormal immune regulation, and impaired angiogenesis, which can be summarized as changes in the wound growth microenvironment. Current clinical treatments include debridement, various functional dressings, growth factors, and antibiotics. While these methods have some efficacy, they still have many limitations, such as the tendency to develop drug resistance, high treatment costs, limited target sites, and unsatisfactory cure rates.

[0004] Traditional Chinese medicine has a long history and rich experience in treating sores and carbuncles. *Callicarpa nudiflora* (Golden Callicarpa) CallicarpanudifloraHook.etArn. This traditional Chinese medicine possesses the functions of clearing heat and detoxifying, astringing and stopping bleeding, and dispersing blood stasis and reducing swelling. It is commonly used to treat external bleeding, burns, and sores. However, scientifically extracting its effective components and formulating an effective compound topical preparation targeting the complex pathogenesis of chronic, refractory wounds (such as damp-heat stagnation and meridian obstruction) still faces many challenges. Therefore, there is an urgent need for a safe, effective, and inexpensive drug to treat and promote the healing of chronic, refractory wounds. Summary of the Invention

[0005] In view of this, the present invention proposes a topical traditional Chinese medicine composition for promoting the healing of chronic and refractory wounds, its preparation method and application, thereby solving the above problems.

[0006] The technical solution of the present invention is as follows: a topical Chinese medicine composition for promoting the healing of chronic and refractory wounds, comprising 1-3 parts of water-soluble lignin obtained by chemical extraction from the stem of Callicarpa nudiflora and 1-3 parts of leaf extract obtained by water extraction from the leaves of Callicarpa nudiflora.

[0007] Preferably, the weight ratio of the external application traditional Chinese medicine composition is: 1 part water-soluble lignin and 2 parts water extract of Callicarpa nudiflora leaves.

[0008] Preferably, the composition further includes pharmaceutically acceptable excipients and is prepared as a topical formulation.

[0009] Preferably, the topical preparation is a gel, hydrogel, ointment, wet dressing, spray, or lotion.

[0010] Preferably, a method for preparing a topical traditional Chinese medicine composition for promoting the healing of chronic, refractory wounds includes the following steps: (1) Extraction of water-soluble lignin: Using the stem of Callicarpa nudiflora as raw material, a compound enzyme was used for enzymatic hydrolysis. The hydrolyzed stem powder was then extracted and purified by co-extraction with a transparent deep eutectic solvent to obtain water-soluble lignin. (2) Preparation of aqueous extract of Callicarpa nudiflora leaves: using Callicarpa nudiflora leaves as raw material, add water and decoct, combine the decoctions, concentrate and obtain an extract in the form of paste; (3) Preparation of the composition: The water-soluble lignin obtained in step (1) and the water extract obtained in step (2) are mixed in the weight ratio described above, and pharmaceutically acceptable excipients are added to prepare a topical preparation.

[0011] Preferably, step (1) specifically includes: 1a) Pretreatment: After freeze-drying the stems of Callicarpa nudiflora at -40~-20℃, the powder is pulverized and passed through a 30-50 mesh sieve; weigh 8-12g of the sieved powder, add 80-120ml of distilled water, adjust the pH to 4.5-5.5, add 0.15-0.25g of compound enzyme, and enzymatically hydrolyze at a constant temperature of 45-55℃ for 1.5-2.5h to destroy the cell wall structure; 1b) Preparation of transparent deep eutectic solvent: Mix choline chloride and lactic acid in a molar ratio of 1.5:1-2.5:1 and stir at 55-65℃ for 20-40 min to obtain transparent deep eutectic solvent (DES); then dilute with distilled water to a mass fraction of 45%-55% to obtain DES extract; 1c) Co-extraction: Mix the enzymatically hydrolyzed stem powder with DES extract at a ratio of 1:12-1:18 (g / mL), place in an ultrasonic-assisted reaction vessel, and extract at a constant temperature of 75-85℃ for 2.5-3.5h, stirring once every 25-35min at 15-25r / min. 1d) Purification: After cooling the extract, centrifuge at 4500-5500 r / min for 10-20 min; take the supernatant and add 2.5-3.5 times the volume of deionized water to precipitate lignin, let stand for 10-14 h and then filter; neutralize the filter residue with 0.8-1.2 mol / L sodium bicarbonate solution to pH 6.5-7.5, and then freeze-dry at -50~-30℃ for 10-14 h to obtain water-soluble lignin.

[0012] Preferably, in step 1a), the mass-to-volume ratio of the naked-flower purple calligraphic stem powder to distilled water is 1:0.8~1.2 (g / mL); the mass ratio of the naked-flower purple calligraphic stem powder to the complex enzyme is 1:0.15-0.25; the complex enzyme is cellulase and laccase in a mass ratio of 3:1, with an enzyme activity ≥5000U / g; and in step 1c), the ultrasonic power of the reaction vessel is 250-350W, and the frequency is 35-45kHz.

[0013] Preferably, step (2) specifically includes: 2a) Decoction: Weigh out the leaves of the naked flower purple callicarpa, soak them in water for 20-40 minutes, heat to boiling and keep simmering for 0.8-1.2 hours, then filter out the decoction; 2b) Repeated decoction: Add water to the dregs again and decoct once more, then combine the two decoctions; 2c) Concentration: The combined decoctions are concentrated by rotary evaporation at 60-80℃ to obtain an extract in the form of a paste.

[0014] Preferably, in steps 2a) and 2b), the amount of water added for the first time is 10-15 times (v / w) the weight of the leaves of Callicarpa nudiflora, and the amount of water added for the second time is 6-10 times (v / w) the weight of the leaves of Callicarpa nudiflora.

[0015] Preferably, the use of the traditional Chinese medicine composition in the preparation of a medicament for promoting wound healing.

[0016] Compared with the prior art, the beneficial effects of the present invention are: The traditional Chinese medicine composition of the present invention consists of two medicinal materials: the stem and leaves of Callicarpa nudiflora.

[0017] Lignin in the traditional Chinese medicine composition is an amorphous biological macromolecule containing a large number of active groups, including phenolic hydroxyl, hydroxyl, and sulfonic acid groups, thus exhibiting good antioxidant activity, free radical scavenging, impact resistance, and tensile properties. Lignin can effectively improve the structural strength of plant cells and resist the invasion of adverse environments. Using lignin as a matrix to carry traditional Chinese medicine components not only helps to exert the efficacy of the extracts, but it can also promote cell proliferation, thereby promoting wound healing. The traditional Chinese medicine composition contains *Callicarpa nudiflora*, which has astringent, hemostatic, heat-clearing, detoxifying, blood-stasis-dispersing, and swelling-reducing effects, precisely targeting the pathogenesis of chronic, intractable wounds caused by exogenous pathogenic factors, damp-heat stagnation, prolonged illness leading to deficiency of vital energy, and blocked meridians. The traditional Chinese medicine composition of this invention astringes and stops bleeding, clears heat and detoxifies, disperses blood stasis and reduces swelling, and promotes cell proliferation in wounds; it is mainly used to treat wounds that are slow to heal due to exogenous pathogenic factors, damp-heat stagnation, and blocked meridians; it has a significant therapeutic effect on chronic, intractable wounds. Attached Figure Description

[0018] Figure 1 The healing process of a chronic, refractory wound after administration of the herbal composition of Example 1 to an animal model.

[0019] Figure 2 The healing process of a chronic, refractory wound after administration of the herbal composition of Example 2 to an animal model.

[0020] Figure 3 The healing process of a chronic, refractory wound after administration of the herbal composition of Example 3 to an animal model. Detailed Implementation

[0021] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0022] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0023] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0024] Example 1 A topical Chinese medicine composition for promoting the healing of chronic, refractory wounds 1) Water-soluble lignin: 1a) Pretreatment: After freeze-drying the stems of Callicarpa nudiflora at -30℃, the powder was crushed and passed through a 40-mesh sieve. 10g of the powder was weighed, 100ml of distilled water was added, the pH was adjusted to 5, 0.2g of compound enzyme (cellulase and laccase = 3:1) was added, and the mixture was kept at 50℃ with shaking for 2h to destroy the cell wall structure. 1b) Preparation of transparent deep eutectic solvent: Choline chloride and lactic acid were mixed in a molar ratio of 2:1 and stirred at 60°C for 30 min to obtain a transparent deep eutectic solvent (DES); then diluted with distilled water to a mass fraction of 50% to obtain DES extract; 1c) Co-extraction: The enzymatically hydrolyzed stem powder and DES extract were mixed at a ratio of 1:15 (g / mL), placed in an ultrasonic-assisted reactor, and extracted at 80℃ for 3 hours with a power of 300W and a frequency of 40kHz, stirring once every 30 minutes at 20r / min. 1d) Purification: After cooling the extract, centrifuge at 5000 r / min for 15 min; take the supernatant and add 3 times the volume of deionized water to precipitate lignin, let stand for 12 h and then filter; neutralize the filter residue with 1 mol / L sodium bicarbonate solution to pH 7, and then freeze dry at -40℃ for 12 h to obtain water-soluble lignin.

[0025] 2) Preparation of Naked Flower Purple Callicarpa Leaf Extract: Weigh 120g of Naked Flower Purple Callicarpa leaves, rinse once with distilled water, soak in 1500ml of water for 30min, boil and keep simmering for 1h, filter out the liquid, add 900ml of water to the residue, boil and keep simmering for 1h, combine the two decoctions, and concentrate into an extract by rotary evaporation at 70℃ and 60r / min.

[0026] 3) Gel Preparation: Add 4g of carbomer 940 to an appropriate amount of distilled water and let it stand overnight to allow it to swell fully. Then, while stirring, add 8g of triethanolamine to prepare the gel matrix. Separately, dissolve 20g of water-soluble lignin, 10g of Callicarpa nudiflora leaf extract, and 0.4g of ethylparaben in 40g of propylene glycol and 120ml of ethanol to prepare the aqueous phase. Add the aqueous phase to the gel matrix while stirring, adding water to a total volume of 400g. After thorough mixing, the herbal composition gel is prepared. Then, pack it into containers, seal it, sterilize it by cobalt-60 irradiation, and store it in a light-proof, sealed container for later use.

[0027] Example 2 The steps are the same as in Example 1. In step (3), 15g of water-soluble lignin and 15g of leaf water extract are prepared.

[0028] Example 3 The steps are the same as in Example 1. In step (3), 10g of water-soluble lignin and 20g of leaf water extract are prepared.

[0029] Experimental example: Pharmacodynamic test 1. Establishment of experimental animals and models SD rats were selected, anesthetized after acclimatization, and fixed after being anesthetized with isoflurane. The hair on the back of the neck of the rats was shaved off with an electric shaver to expose the skin on the back of the neck. The skin was then disinfected with cotton balls moistened with 75% alcohol, and a wound with a diameter of 1.8 cm was created on the skin on the back of the neck.

[0030] In the model group and the compound group, rats were injected intramuscularly around the wound to create a non-healing wound. In the normal control group, no hydrocortisone injection was given.

[0031] Each rat was administered intramuscular penicillin (4000 units / rat) for four consecutive days. After injection, the rats were kept warm until they regained consciousness, and then housed individually with ample feed and water. 2. Grouping and Dosing After successful model creation, the models are randomly divided into: Model control group: Blank gel matrix was applied to the wound.

[0032] Traditional Chinese medicine composition group: Apply the gels prepared in Examples 1, 2 and 3 respectively.

[0033] Apply once daily, after disinfection. Remove scabs every other day: First, moisten the scabs on the wound with physiological saline, then anesthetize the rats with isoflurane, and gently peel off the scabs with sterilized instruments. After peeling, disinfect the wound with povidone-iodine, and after the disinfectant has dried, apply gel to the wound. Observe regularly.

[0034] 3. Experimental Results The wound area was measured on days 0, 7, and 14 after drug administration, and the healing rate was calculated. The results are shown in Table 1.

[0035] Table 1: Wound healing status in rats in Example 1 (wound area, cm²) 2 (x±s, n=8)

[0036] ** indicates that compared with the model control, P < 0.01.

[0037] Table 2: Wound healing status in rats in Example 2 (wound area, cm²) 2 (x±s, n=8)

[0038] * indicates that P < 0.05 compared to the control group.

[0039] Table 3: Wound healing status in rats in Example 3 (wound area, cm²) 2 (x±s, n=8)

[0040] ** indicates that compared with the model control, P < 0.01.

[0041] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An external use traditional Chinese medicine composition for promoting the healing of chronic refractory wounds, characterized in that, The active ingredients are composed of 1-3 parts of water-soluble lignin obtained by chemical extraction from the stems of S. nudiflora and 1-3 parts of leaf extract obtained by water extraction from the leaves of S. nudiflora.

2. The external use traditional Chinese medicine composition for promoting the healing of chronic refractory wounds according to claim 1, characterized in that, The raw materials are composed of 1 part of water-soluble lignin and 2 parts of water extract of S. nudiflora leaves.

3. The external use traditional Chinese medicine composition for promoting the healing of chronic refractory wounds according to claim 1 or 2, characterized in that, The composition further comprises pharmaceutically acceptable excipients and is prepared into a topical preparation.

4. The external use traditional Chinese medicine composition for promoting the healing of chronic refractory wounds according to claim 3, characterized in that, The topical preparation is a gel, a hydrogel, a paste, a wet compress, a spray or a lotion.

5. The method for preparing the external use traditional Chinese medicine composition for promoting the healing of chronic refractory wounds according to claim 1 or 2, characterized in that, The method comprises the following steps: (1) Extraction of water-soluble lignin: S. nudiflora stems are used as raw materials, and a composite enzyme is used for enzymatic hydrolysis. The stem powder after enzymatic hydrolysis is co-extracted with a transparent deep eutectic solvent, purified, and water-soluble lignin is obtained. (2) Preparation of water extract of S. nudiflora leaves: S. nudiflora leaves are used as raw materials, and water is added for decoction. The decoction is combined and concentrated to obtain the water extract in the form of extract. (3) Preparation of the composition: the water-soluble lignin obtained in step (1) and the water extract obtained in step (2) are mixed according to the weight ratio, and pharmaceutically acceptable excipients are added to prepare a topical preparation.

6. The method for preparing a topical traditional Chinese medicine composition for promoting the healing of chronic, refractory wounds as described in claim 5, characterized in that, Step (1) specifically comprises: 1a) Pretreatment: S. nudiflora stems are freeze-dried at -40~-20℃, crushed, and passed through a 30-50 mesh sieve. The sieved powder is weighed, distilled water is added, the pH is adjusted to 4.5-5.5, 0.15-0.25g of a composite enzyme is added, and the cell wall structure is destroyed by constant temperature oscillation enzymolysis at 45-55℃ for 1.5-2.5h; 1b) Preparation of transparent deep eutectic solvent: choline chloride and lactic acid are mixed at a molar ratio of 1.5:1-2.5:1, stirred at 55-65℃ for 20-40min to prepare a transparent deep eutectic solvent (DES), and then diluted with distilled water to a mass fraction of 45%-55% to obtain a DES extraction solution; 1c) Co-extraction: the stem powder after enzymatic hydrolysis is mixed with the DES extraction solution at a ratio of 1:12-1:18 (g / mL), placed in an ultrasonic-assisted reaction kettle, and extracted at a constant temperature of 75-85℃ for 2.5-3.5h. During the extraction, the mixture is stirred at 15-25r / min every 25-35min; 1d) Purification: the extraction solution is cooled and centrifuged at 4500-5500r / min for 10-20min. The supernatant is precipitated with 2.5-3.5 times the volume of deionized water, and the lignin is precipitated. After standing for 10-14h, the filtrate is filtered. The residue is neutralized with 0.8-1.2mol / L sodium bicarbonate solution to pH 6.5-7.5, and then freeze-dried at -50~-30℃ for 10-14h to obtain water-soluble lignin.

7. The method for preparing a topical traditional Chinese medicine composition for promoting the healing of chronic, refractory wounds as described in claim 6, characterized in that, In step 1a), the mass / volume ratio g / mL of the S. nudiflora stem powder and distilled water is 1:0.8~1.2; the mass ratio of the S. nudiflora stem powder and the composite enzyme is 1:0.15-0.25, and the composite enzyme is cellulase and laccase at a mass ratio of 3:1, with an enzyme activity of ≥5000U / g. In step 1c), the ultrasonic power of the reaction kettle is 250-350W, and the frequency is 35-45kHz.

8. The method for preparing a topical traditional Chinese medicine composition for promoting the healing of chronic, refractory wounds as described in claim 5, characterized in that, Step (2) specifically comprises: 2a) Decoction: S. nudiflora leaves are weighed, soaked in water for 20-40min, heated to boiling, and kept at a low boil for 0.8-1.2h, and the decoction is filtered out; 2b) Repeated decoction: water was added to the residue again, and repeated decoction was performed once, and the two decoctions were combined; 2c) Concentration: the combined decoction was concentrated by rotary evaporation at 60-80°C to obtain the extract of water extract.

9. The external use traditional Chinese medicine composition for promoting the healing of chronic refractory wounds according to claim 8, characterized in that, In steps 2a) and 2b), the amount of water added for the first time was 10-15 times the weight of the leaves of Ligustrum robustum (v / w), and the amount of water added for the second time was 6-10 times the weight of the leaves of Ligustrum robustum (v / w).

10. Use of the traditional Chinese medicine composition of any one of claims 1-4 in the preparation of a medicament for promoting wound healing.

Images