Pharmaceutical composition for treating burns and scalds in intensive care and preparation method thereof

Abstract

The invention provides a pharmaceutical composition for treating burns and scalds in intensive care and a preparation method thereof, and belongs to the technical field of medicines. Comprising the following raw materials in parts by weight: 1-1.5 parts of analgesic temperature response liposome, 0.5-1 part of co-loaded exosome, 10-15 parts of lithospermum erythrorhizon, 5-10 parts of astragalus membranaceus, 3-7 parts of rheum officinale, 3-5 parts of radix angelicae, 3-5 parts of angelica sinensis, 5-10 parts of pseudo-ginseng, 10-15 parts of chitosan and 20-25 parts of sodium beta-glycerophosphate. According to the invention, the transdermal absorption of the medicine is greatly improved, the intelligent administration is realized, the side effect is reduced, the effect is better, the pain of a patient is reduced, and the system is suitable for healing various types of burn and scald wounds and has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a pharmaceutical composition for treating burns in intensive care and its preparation method. Background Technology

[0002] While the causes of burns and scalds vary, the principles of treatment remain consistent, addressing key issues such as wound pain, progressive necrosis, susceptibility to infection, and scar healing. With societal development, the theoretical level and clinical experience of traditional Chinese medicine in treating burns and scalds have improved, resulting in some progress in this field.

[0003] Ancient Chinese medicine literature contains records of burn treatment, such as *Fifty-Two Prescriptions*, *Introduction to Medicine: Burns*, *Complete Collection of Ulcer Treatments*, and *Ghostly Prescriptions*. Chen Shiduo's *Dongtian Aozhi* from the Qing Dynasty states, "To treat burns, both internal and external treatments must be applied simultaneously, so that the fire toxin can be easily resolved." This already established the principle of treating burns from both internal and external sources. Based on the experience of treating burns in ancient and modern medicine, modern Chinese medicine has summarized five major treatment principles: clearing heat and detoxifying, nourishing yin and generating fluids, tonifying qi and regulating the spleen, promoting blood circulation and removing blood stasis, and supporting the body and draining pus. Burn treatment is mainly divided into two types: dry therapy and wet therapy. Western medicine often uses dry therapy for burns, which mainly relies on interfering with external factors, using antibiotics, and surgery. First, the wound is dried and scabs formed, then surgically removed and skin grafted. Wet therapy, on the other hand, is widely used in traditional Chinese medicine for burn treatment. External application of Chinese medicine and its preparations creates a suitable humidity-controlled pharmacological environment, liquefying necrotic tissue and preserving regenerative tissue from a physiological regeneration and repair perspective, allowing the wound to heal spontaneously. However, traditional Western or Chinese medicine treatments for burns are characterized by severe pain, slow and poor efficacy, and a high likelihood of leaving scars.

[0004] Chinese Patent (CN101884709A) discloses a medicated ointment for treating burns, made from frankincense, myrrh, catechu, dragon's blood, safflower, dragon bone, beeswax, elephant skin, coptis, rhubarb, and sesame oil. This invention can promote blood circulation, remove blood stasis, astringe, and relieve pain, thereby achieving the purpose of treating burns. Chinese Patent (CN107714781A) discloses a pharmaceutical composition for treating burns, consisting of two functional components, a and b. Component a clears heat and detoxifies, reduces inflammation and relieves pain; its ingredients are 10-40 parts of Polygonum cuspidatum, 5-35 parts of dandelion, 5-35 parts of Lithospermum erythrorhizon, and 1-20 parts of borneol. Component b promotes blood circulation, removes blood stasis, and promotes tissue regeneration; its ingredients are 10-40 parts of rhubarb, 10-40 parts of Sanguisorba officinalis, 1-20 parts of Codonopsis pilosula, 1-20 parts of Astragalus membranaceus, and 50-200 parts of egg white. Using different Chinese herbal medicines according to their different effects can fully utilize the synergistic effects of each herb component, resulting in more significant and stable efficacy compared to single-herb administration. It is mainly used for early and mid-stage burns characterized by redness, swelling, heat, pain, and significant exudation, reducing wound infection, accelerating wound healing, and effectively decreasing the formation of ulcers at the burn site. However, existing single-herb burn medications still have limitations in efficacy, exhibiting problems such as long treatment time, poor results, and a tendency to leave scars. Summary of the Invention

[0005] The purpose of this invention is to propose a pharmaceutical composition for treating burns in intensive care and its preparation method. It has the advantages of promoting wound healing, reducing scar formation, synergistic repair, greatly improving transdermal drug absorption, realizing intelligent drug delivery, reducing side effects, improving efficacy, reducing patient suffering, and is suitable for healing various types of burn wounds, with broad application prospects.

[0006] The technical solution of this invention is implemented as follows: This invention provides a pharmaceutical composition for treating burns in intensive care, comprising the following raw materials in parts by weight: 1-1.5 parts of analgesic temperature-responsive liposomes, 0.5-1 parts of co-loaded exosomes, 10-15 parts of Lithospermum erythrorhizon, 5-10 parts of Astragalus membranaceus, 3-7 parts of Rheum palmatum, 3-5 parts of Angelica dahurica, 3-5 parts of Angelica sinensis, 5-10 parts of Panax notoginseng, 10-15 parts of chitosan, and 20-25 parts of sodium β-glycerophosphate.

[0007] As a further improvement of the present invention, the preparation of the analgesic temperature-responsive liposomes is as follows: S1. Ketamine and sufentanil were dissolved in hot ethanol, cooled to room temperature, and evaporated in an open container to obtain a eutectic. The eutectic was then filtered, washed, dried, and pulverized to obtain the ketamine and sufentanil eutectic. S2. Dipalmitoylphosphatidylcholine, cholesterol, ketamine and sufentanil cocrystal were added to diethyl ether, then added dropwise to buffer solution, stirred and mixed, the diethyl ether was removed by heating under reduced pressure, sonicated and filtered to obtain analgesic temperature-responsive liposomes.

[0008] Sufentanil is an N-4 thiophene derivative of fentanyl. It is a specific μ-opioid receptor agonist with good analgesic effects, rapid onset of action, and long duration of analgesia. However, its strong lipophilicity and large molecular weight greatly limit its transdermal absorption efficiency. High doses can cause adverse reactions such as nausea, vomiting, and respiratory depression, which limits its application to some extent.

[0009] Ketamine is commonly used as an intravenous anesthetic, possessing a definite, short-acting analgesic effect and the ability to provide dissociative anesthesia. It has a very high affinity for NMDA receptors, primarily antagonizing them by reducing voltage, the mean opening time of ligand-gated channels, and their opening frequency. Secondly, ketamine can promote the effective release of endogenous opioid peptides and inhibit the reuptake of neurotransmitters such as dopamine and norepinephrine, thus producing an analgesic effect. During inflammatory responses to traumatic stress such as burns, the number of NMDA receptors increases, exacerbating the inflammatory stress and leading to prolonged pain sensitization in the pain center. Ketamine can both excite the brainstem and limbic system and block the transmission of pain impulses to the thalamus and neocortex, thus inhibiting hyperalgesia. However, high doses can easily cause adverse reactions such as amnesia, delirium, and hallucinations, and its large molecular weight limits its transdermal absorption efficiency to some extent.

[0010] Therefore, the combined use of ketamine and sufentanil yields good analgesic effects, and preparing them in co-crystal form better achieves simultaneous release, synergistic peak absorption, and synergistic effects. However, conventional chemical penetration enhancers are difficult to promote the synergistic transdermal absorption of ketamine and sufentanil, resulting in poor efficacy. Therefore, the inventors of this invention employ a liposome encapsulation method. On the one hand, by partially fusing with the outer surface of the stratum corneum, drug permeability is increased, thus solving the problem of transdermal absorption efficiency for large molecular weight drugs. On the other hand, liposomes have sustained-release properties, ensuring continuous drug release and controlling the release rate, avoiding side effects from excessive drug release. Furthermore, liposomes can protect the encapsulated drug from premature degradation, are safe and non-irritating, and improve the solubility of both drugs, thereby increasing the transdermal drug volume and rate. However, since analgesics are not needed throughout the treatment of burn wounds, only in the acute initial stage of the wound, and not in large quantities during the later healing and scab formation period, optimizing the structure of liposomes to regulate the release of analgesics has become one of the problems that needs to be solved. The inventors later discovered that in the acute early stage of a wound, the skin temperature rises locally due to a large amount of inflammatory response. Therefore, the inventors designed a temperature-responsive liposome that effectively solved the above problem.

[0011] The temperature-responsive liposomes prepared by this invention release little drug at temperatures below 36°C, but release a large amount of drug at 37-40°C, thereby achieving rapid drug release in the acute phase of the wound to exert analgesic and anesthetic effects. The drug release rate decreases in the later stage, thus adapting well to various stages of wound healing and greatly reducing the patient's pain.

[0012] As a further improvement of the present invention, the mass ratio of ketamine and sufentanil in step S1 is 40-50:1, and the temperature of the hot ethanol is 60-70℃; the mass ratio of dipalmitoylphosphatidylcholine, cholesterol, ketamine and sufentanil cocrystal in step S2 is 5-8:1-3:0.8-1.2, the buffer solution is PBS buffer with pH=7.4, the sonication time is 15-30 min, and the filtration is sequential filtration through 0.45μm and 0.22μm organic membranes.

[0013] As a further improvement of the present invention, the method for preparing the co-loaded exosomes is as follows: T1. Centrifuge to remove exosomes from fetal bovine serum, prepare a culture medium for culturing deer antler stem cells, collect the supernatant after the cells have grown to confluence, and collect the deer antler stem cell exosomes by gradient centrifugation; T2. Dissolve salvianolic acid B in buffer solution, add deer antler stem cell exosomes, incubate, dialyze, and obtain the first drug-loaded exosomes; T3. Tanshinone IIA was dissolved in DMSO, mixed with an aqueous solution of the first drug-loaded exosomes, placed under an electroporator, electrolyzed, removed, centrifuged, and freeze-dried to obtain co-loaded exosomes.

[0014] Deer antlers can fully regenerate every year. The healing of wounds on the antler stalks differs from typical scar healing. It involves the regeneration of skin appendages, including hair follicles and sebaceous glands, and the collagen arrangement closely resembles the basket-like pattern of normal skin, classifying it as regenerative healing. Deer antler regeneration is a stem cell-based process, known as antler stem cells. However, stem cells suffer from inconvenient storage and low activity stability. In contrast, stem cell-derived exosomes offer advantages in treating skin wounds, including immunocompatibility, high stability, no tumorigenic risk, convenient storage, and ease of quantitative use. Furthermore, active ingredients in Danshen (Salvia miltiorrhiza), such as tanshinone IIA and salvianolic acid B, can improve scar formation through mechanisms like anti-fibrosis, regulation of inflammatory responses, and promotion of regeneration. However, both suffer from poor stability and low oral bioavailability. By combining several technologies and leveraging their strengths while mitigating their weaknesses, the inventors of this invention extracted exosomes from deer antler stem cells and used them to load tanshinone IIA and salvianolic acid B. This not only solved the problems of poor stability and low oral bioavailability of tanshinone IIA and salvianolic acid B, but also demonstrated that the co-loaded exosomes have good transdermal drug delivery and targeted drug delivery characteristics. They release the exosomes in a responsive manner in the wound microenvironment, reducing side effects. This not only effectively promotes wound healing but also better reduces scar formation, exhibiting a synergistic repair effect.

[0015] As a further improvement of the present invention, the mass ratio of salvianolic acid B, tanshinone IIA, and deer antler stem cell exosomes is 1-2:0.5-1:15-20; the culture medium in step T1 is DMEM complete culture medium containing 8-12% fetal bovine serum, 0.5-1.5wt% penicillin, and 0.5-1.5wt% streptomycin, and the deer antler stem cells are passage 5-7; the incubation temperature in step T2 is 36-38℃, the incubation time is 1-3h, the buffer is PBS buffer with pH=7.4, and the protein content of the first drug-loaded exosome aqueous solution is 0.3-0.4mg / mL; the number of electroshocks in step T3 is 3-7 times, and the voltage is 0.5-1V.

[0016] The present invention further protects a method for preparing the above-mentioned pharmaceutical composition for treating burns in intensive care, comprising the following steps: (1) Mix Lithospermum erythrorhizon, Astragalus membranaceus, Rheum palmatum, Angelica dahurica, Angelica sinensis and Panax notoginseng, pulverize and sieve to obtain Chinese medicine powder. Add the Chinese medicine powder to ethyl acetate, heat and reflux to extract, filter, add emulsifier F68 and 1,2-propanediol to the filtrate to form an emulsion, add sodium alginate solution dropwise to emulsify, add petroleum ether to form a double emulsion, add calcium chloride solution dropwise to crosslink, centrifuge, wash and dry to obtain Chinese medicine microspheres; (2) Dissolve chitosan in acid to obtain chitosan solution, thus obtaining solution A; (3) Dissolve sodium β-glycerophosphate in sodium bicarbonate solution to obtain solution B; (4) Add solution B to solution A, add traditional Chinese medicine microspheres, analgesic temperature-responsive liposomes and co-loaded exosomes, stir and mix evenly to obtain a drug composition for treating burns in intensive care.

[0017] The extracts of Lithospermum erythrorhizon, Astragalus membranaceus, Rheum palmatum, Angelica dahurica, Angelica sinensis, and Panax notoginseng, obtained through ethyl acetate extraction, yielded a wealth of effective antibacterial, anti-inflammatory, and wound-healing components, which achieved good effects in preventing wound infection, reducing inflammatory response, stopping bleeding, and promoting wound healing.

[0018] This invention uses chitosan to prepare a temperature-sensitive hydrogel. Chitosan has good antibacterial effects and can synergistically improve the anti-infection effect of wounds. At the same time, under liquid conditions, it has a good retention effect on temperature-responsive liposomes and co-loaded exosomes of analgesics, avoiding their inactivation and improving stability. At body temperature of 37°C, it forms a gel state, prolonging the administration time, which can better promote wound healing, reduce scar formation, and has few side effects and synergistic repair.

[0019] As a further improvement of the present invention, the heating and reflux extraction time in step (1) is 4-8h, the solid-liquid ratio of the Chinese herbal powder and ethyl acetate is 1:5-10g / mL, the concentration of the sodium alginate solution is 2-4wt%, the concentration of the calcium chloride solution is 1-3wt%, and the crosslinking time is 4-6h.

[0020] As a further improvement of the present invention, the acid solution in step (2) is a 1-3 wt% hydrochloric acid or acetic acid solution, and the concentration of the chitosan solution is 3-5 wt%.

[0021] As a further improvement of the present invention, the concentration of sodium β-glycerophosphate in solution B in step (3) is 50-60 wt%, and the concentration of sodium bicarbonate solution is 0.3-0.5 mol / L.

[0022] As a further improvement of the present invention, the pharmaceutical composition for treating burns in the intensive care unit described in step (4) is liquid at room temperature and forms a gel at 37°C after being applied to the skin.

[0023] The present invention has the following beneficial effects: 1. The raw materials of this invention are widely available, the preparation method is simple, the storage stability is good, the administration time is long, and it has a good effect on promoting wound healing, reducing scar formation, and synergistic repair.

[0024] 2. This invention overcomes the application defects of raw materials in terms of transdermal absorption and stability, greatly improves the transdermal absorption of drugs, realizes intelligent drug delivery, reduces side effects, has better effects, reduces patient suffering, is suitable for healing various types of burns and scalds, and has broad application prospects. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0026] Figure 1 DSC curves of the analgesic temperature-responsive liposomes prepared in Example 3.

[0027] Figure 2 TEM image of deer antler stem cell exosomes prepared in step T1 of Example 4. Detailed Implementation

[0028] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0029] Preparation Example 1: Preparation of analgesic temperature-responsive liposomes: S1. Dissolve 0.4g ketamine and 10mg sufentanil in 50mL of hot ethanol (temperature 60-70℃), cool to room temperature, evaporate in an open container to obtain a eutectic, filter, wash, dry, and pulverize to obtain a ketamine and sufentanil eutectic; S2. Add 0.5g dipalmitoylphosphatidylcholine, 0.1g cholesterol, 80mg ketamine and sufentanil cocrystal to 50mL of diethyl ether, add dropwise to 100mL of PBS buffer (pH=7.4), stir and mix, remove the diethyl ether by rotary evaporation under reduced pressure, sonicate for 15min, filter through 0.45μm and 0.22μm organic membranes sequentially, dry, and obtain analgesic temperature-responsive liposomes.

[0030] Preparation Example 2: Preparation of analgesic temperature-responsive liposomes: S1. Dissolve 0.5g ketamine and 10mg sufentanil in 50mL of hot ethanol (temperature 60-70℃), cool to room temperature, evaporate in an open container to obtain eutectic, filter, wash, dry, and pulverize to obtain ketamine and sufentanil eutectic; S2. Add 0.8g dipalmitoylphosphatidylcholine, 0.3g cholesterol, 120mg ketamine and sufentanil cocrystal to 50mL of diethyl ether, then add dropwise to 100mL of PBS buffer (pH=7.4), stir and mix, remove the diethyl ether by rotary evaporation under reduced pressure, sonicate for 30min, filter through 0.45μm and 0.22μm organic membranes sequentially, and dry to obtain analgesic temperature-responsive liposomes.

[0031] Preparation Example 3: Preparation of analgesic temperature-responsive liposomes: S1. Dissolve 0.45g ketamine and 10mg sufentanil in 50mL of hot ethanol (temperature 60-70℃), cool to room temperature, evaporate in an open container to obtain eutectic, filter, wash, dry, and pulverize to obtain ketamine and sufentanil eutectic; S2. 0.65 g dipalmitoylphosphatidylcholine, 0.2 g cholesterol, 100 mg ketamine, and sufentanil cocrystal were added to 50 mL of diethyl ether. This mixture was then added dropwise to 100 mL of PBS buffer (pH 7.4), stirred, and the ether was removed by rotary evaporation under reduced pressure. The mixture was sonicated for 20 min, filtered sequentially through 0.45 μm and 0.22 μm organic membranes, and dried to obtain analgesic temperature-responsive liposomes. 5-10 mg of the analgesic temperature-responsive liposomes were weighed and spread on the bottom of an aluminum crucible. The mixture was scanned within a heating range of 20-60 °C at a heating rate of 5 °C / min, repeated twice, and the DSC curve was measured. A blank crucible was used as a control. The DSC curve is shown below. Figure 1 As shown in the figure, there are two absorption peaks at 37.8 and 41.2℃, indicating obvious temperature sensitivity. The test results show that the main range of its temperature sensitivity is 38-42℃.

[0032] Comparative Preparation Example 1 The only difference from Preparation Example 3 is that dipalmitoylphosphatidylcholine is replaced by an equal mass of soybean lecithin.

[0033] Comparative Preparation Example 2 The only difference from Preparation Example 3 is that ketamine was not added in step S1, and the ketamine and sufentanil eutectic was replaced by an equal mass of sufentanil.

[0034] Comparative preparation example 3 The only difference from Preparation Example 3 is that sufentanil was not added in step S1, and the ketamine and sufentanil cocrystal were replaced by an equal mass of ketamine.

[0035] Preparation Example 4: Preparation of co-loaded exosomes: T1. 100000× g Centrifuge for 2 hours to remove exosomes from fetal bovine serum, prepare culture medium for culturing 6th generation deer antler stem cells, and collect the supernatant after the cells have reached confluence. (The solution is then centrifuged at 300×...) gCentrifuge for 10 minutes to remove dead cells; 2000× g Centrifuge for 15 minutes to remove cell debris; 10000× g Centrifuge for 30 min to remove smaller cell debris; sterilize by filtration through a 0.22 μm membrane; 100,000 × 10⁻⁶ g Centrifuge for 90 min, discard the liquid in the tube, freeze dry, and obtain deer antler stem cell exosomes; Figure 2 The image shows a TEM image of the exosomes obtained from deer antler stem cells. As can be seen from the image, the exosomes are round or elliptical bilayer vesicle-like structures with a particle size ranging from 50 to 150 nm.

[0036] The culture medium is a DMEM complete medium containing 10% fetal bovine serum, 1 wt% penicillin, and 1 wt% streptomycin. T2. Dissolve 0.1g of salvianolic acid B in 100mL of PBS buffer (pH=7.4), add 1.5g of deer antler stem cell exosomes, incubate at 37℃ for 2h, dialyze to obtain the first drug-loaded exosomes, and adjust the concentration to a protein content of 0.3mg / mL. T3. Dissolve 0.5g of tanshinone IIA in 10mL of DMSO, mix with the aqueous solution of the first drug-loaded exosomes prepared in step T2, place under an electroporator, electroporate 7 times at a voltage of 0.5V, remove, centrifuge, freeze dry, and obtain co-loaded exosomes.

[0037] Preparation Example 5: Preparation of co-loaded exosomes: T1. 100000× g Centrifuge for 2 hours to remove exosomes from fetal bovine serum, prepare culture medium for culturing 6th generation deer antler stem cells, and collect the supernatant after the cells have reached confluence. (The solution is then centrifuged at 300×...) g Centrifuge for 10 minutes to remove dead cells; 2000× g Centrifuge for 15 minutes to remove cell debris; 10000× g Centrifuge for 30 min to remove smaller cell debris; sterilize by filtration through a 0.22 μm membrane; 100,000× g Centrifuge for 90 min, discard the liquid in the tube, freeze dry, and obtain deer antler stem cell exosomes; The culture medium is a DMEM complete medium containing 10% fetal bovine serum, 1 wt% penicillin, and 1 wt% streptomycin. T2. Dissolve 0.2g of salvianolic acid B in 100mL of PBS buffer (pH=7.4), add 2g of deer antler stem cell exosomes, incubate at 37℃ for 2h, dialyze to obtain the first drug-loaded exosomes, and adjust the concentration to a protein content of 0.4mg / mL. T3. Dissolve 1g of tanshinone IIA in 10mL of DMSO, mix with the aqueous solution of the first drug-loaded exosomes prepared in step T2, place under an electroporator, electroporate 3 times at a voltage of 1V, remove, centrifuge, freeze dry, and obtain co-loaded exosomes.

[0038] Preparation Example 6: Preparation of co-loaded exosomes: T1. 100000× g Centrifuge for 2 hours to remove exosomes from fetal bovine serum, prepare culture medium for culturing 6th generation deer antler stem cells, and collect the supernatant after the cells have reached confluence. (The solution is then centrifuged at 300×...) g Centrifuge for 10 minutes to remove dead cells; 2000× g Centrifuge for 15 minutes to remove cell debris; 10000× g Centrifuge for 30 min to remove smaller cell debris; sterilize by filtration through a 0.22 μm membrane; 100,000× g Centrifuge for 90 min, discard the liquid in the tube, freeze dry, and obtain deer antler stem cell exosomes; The culture medium is a DMEM complete medium containing 10% fetal bovine serum, 1 wt% penicillin, and 1 wt% streptomycin. T2. Dissolve 0.15g of salvianolic acid B in 100mL of PBS buffer (pH=7.4), add 1.7g of deer antler stem cell exosomes, incubate at 37℃ for 2h, dialyze to obtain the first drug-loaded exosomes, and adjust the concentration to a protein content of 0.35mg / mL. T3. Dissolve 0.7g of tanshinone IIA in 10mL of DMSO, mix with the aqueous solution of the first drug-loaded exosomes prepared in step T2, place under an electroporator, electroporate 5 times at a voltage of 0.7V, remove, centrifuge, freeze dry, and obtain co-loaded exosomes.

[0039] Comparative preparation example 4 The only difference from Preparation Example 6 is that step T2 was not performed, and in step T3, the first drug-loaded exosome was replaced by deer antler stem cell exosomes.

[0040] Comparative preparation example 5 The only difference from Preparation Example 6 is that step T3 was not performed, and the first drug-loaded exosome is a co-loaded exosome.

[0041] Comparative preparation example 6 The difference from Preparation Example 6 is that the co-loaded exosomes are replaced by a mixture, which is prepared as follows: A mixture was prepared by simply mixing 0.15g of salvianolic acid B, 0.7g of tanshinone IIA, and 1.7g of deer antler stem cell exosomes.

[0042] Example 1 This embodiment provides a method for preparing a pharmaceutical composition for treating burns in intensive care, comprising the following steps: (1) Mix 10g of Lithospermum erythrorhizon, 5g of Astragalus membranaceus, 3g of Rheum palmatum, 3g of Angelica dahurica, 3g of Angelica sinensis and 5g of Panax notoginseng, pulverize and sieve to obtain Chinese medicine powder. Add the Chinese medicine powder to ethyl acetate. The solid-liquid ratio of Chinese medicine powder to ethyl acetate is 1:5g / mL. Heat and reflux for 4h. Filter. Add 0.5g of emulsifier F68 and 1g of 1,2-propanediol to the filtrate to form an emulsion. Add 200mL of 2wt% sodium alginate solution dropwise. Emulsify at 5000r / min for 15min. Add 500mL of petroleum ether to form a double emulsion. Add 10mL of 1wt% calcium chloride solution dropwise. Crosslink for 4h. Centrifuge, wash and dry to obtain Chinese medicine microspheres. (2) Dissolve 10g of chitosan in a 2wt% acetic acid solution to obtain solution A with a concentration of 3wt%; (3) Dissolve 20g of sodium β-glycerophosphate in a 0.3mol / L sodium bicarbonate solution to obtain solution B, where the concentration of sodium β-glycerophosphate is 50wt%. (4) Add solution B to solution A, add the Chinese medicine microspheres prepared in step (1), 1g of analgesic temperature-responsive liposomes prepared in preparation example 1, and 0.5g of co-loaded exosomes prepared in preparation example 4, stir and mix for 30 minutes, adjust the pH to 7, and obtain a drug composition for treating burns in intensive care. It is liquid at room temperature and forms a gel at 37°C after being applied to the skin.

[0043] Example 2 This embodiment provides a method for preparing a pharmaceutical composition for treating burns in intensive care, comprising the following steps: (1) Mix 15g of Lithospermum erythrorhizon, 10g of Astragalus membranaceus, 7g of Rheum palmatum, 5g of Angelica dahurica, 5g of Angelica sinensis, and 10g of Panax notoginseng, pulverize, and sieve to obtain Chinese medicine powder. Add the Chinese medicine powder to ethyl acetate, with a solid-liquid ratio of 1:10g / mL. Heat and reflux for 8h, filter, filter again, add 0.5g of emulsifier F68 and 1g of 1,2-propanediol to the filtrate to form an emulsion, dropwise add 200mL of 4wt% sodium alginate solution, emulsify at 5000r / min for 15min, add 500mL of petroleum ether to form a double emulsion, dropwise add 10mL of 3wt% calcium chloride solution, crosslink for 6h, centrifuge, wash, dry, and obtain Chinese medicine microspheres; (2) Dissolve 15g of chitosan in a 2wt% acetic acid solution to obtain solution A with a concentration of 5wt%; (3) Dissolve 25g of sodium β-glycerophosphate in a 0.5mol / L sodium bicarbonate solution to obtain solution B, with a sodium β-glycerophosphate concentration of 60wt%; (4) Add solution B to solution A, add the traditional Chinese medicine microspheres prepared in step (1), 1.5g of analgesic temperature-responsive liposomes prepared in preparation example 2, and 1g of co-loaded exosomes prepared in preparation example 5. Stir and mix for 30 minutes, adjust the pH to 7, and obtain a drug composition for treating burns in intensive care. It is liquid at room temperature and forms a gel at 37°C after being applied to the skin.

[0044] Example 3 This embodiment provides a method for preparing a pharmaceutical composition for treating burns in intensive care, comprising the following steps: (1) Mix 12g of Lithospermum erythrorhizon, 7g of Astragalus membranaceus, 5g of Rheum palmatum, 4g of Angelica dahurica, 4g of Angelica sinensis, and 7g of Panax notoginseng, pulverize, and sieve to obtain Chinese medicine powder. Add the Chinese medicine powder to ethyl acetate. The solid-liquid ratio of Chinese medicine powder to ethyl acetate is 1:8g / mL. Heat and reflux for 6h, filter, filter again, add 0.5g of emulsifier F68 and 1g of 1,2-propanediol to the filtrate to form an emulsion, dropwise add 200mL of 3wt% sodium alginate solution, emulsify at 5000r / min for 15min, add 500mL of petroleum ether to form a double emulsion, dropwise add 10mL of 3wt% calcium chloride solution, crosslink for 5h, centrifuge, wash, dry, and obtain Chinese medicine microspheres; (2) Dissolve 12g of chitosan in a 2wt% acetic acid solution to obtain solution A with a concentration of 4wt%; (3) Dissolve 22g of sodium β-glycerophosphate in a 0.4mol / L sodium bicarbonate solution to obtain solution B, with a sodium β-glycerophosphate concentration of 56wt%. (4) Add solution B to solution A, add the traditional Chinese medicine microspheres prepared in step (1), 1.2g of analgesic temperature-responsive liposomes prepared in preparation example 3, and 0.7g of co-loaded exosomes prepared in preparation example 6. Stir and mix for 30 minutes, adjust the pH to 7, and obtain a drug composition for treating burns in intensive care. It is liquid at room temperature and forms a gel at 37°C after being applied to the skin.

[0045] Comparative Example 1 The only difference from Example 3 is that the analgesic temperature-responsive liposomes were prepared by Comparative Preparation Example 1.

[0046] Comparative Example 2 The only difference from Example 3 is that the analgesic temperature-responsive liposomes were prepared by Comparative Preparation Example 2.

[0047] Comparative Example 3 The only difference from Example 3 is that the analgesic temperature-responsive liposomes were prepared in Comparative Preparation Example 3.

[0048] Comparative Example 4 The only difference from Example 3 is that the co-loaded exosomes were prepared by Comparative Preparation Example 4.

[0049] Comparative Example 5 The only difference from Example 3 is that the co-loaded exosomes were prepared from Comparative Preparation Example 5.

[0050] Comparative Example 6 The only difference from Example 3 is that the co-loaded exosomes were prepared by Comparative Preparation Example 6.

[0051] Test Example 1: Hot Plate Test Healthy female Kunming mice weighing 20-23g were selected and placed one by one on a 55°C hot plate. The normal pain threshold of each mouse was measured: the latency from when the mouse was placed on the hot plate until it licked its hind paw was defined as the normal pain threshold (s). After a 30-minute interval, the measurement was repeated, and the average of the two pain threshold measurements was taken as the normal pain threshold for each mouse before drug administration. Mice with latency times less than 5s or greater than 30s, or those that jumped, were excluded. Ten mice were randomly divided into seven groups of ten mice each. The blank group was coated with 1 mL of physiological saline per mouse, while the experimental groups were coated with 1 mL of the drug composition for treating burns in intensive care prepared in Examples 1-3 or Comparative Examples 1-3, respectively. The drugs were administered once daily for two consecutive days. The pain threshold of each mouse was measured 30 minutes after the last administration. If a mouse did not lick its hind paw after 60 seconds on the hot plate, it was immediately removed, and the result was recorded as 60 seconds. The results are shown in Table 1.

[0052] Table 1

[0053] Note: * indicates P < 0.05 compared to the control group.

[0054] As shown in Table 1, the drug compositions for treating burns in intensive care prepared in Examples 1-3 can significantly delay the time mice lick their hind paws, increase the pain threshold of mice, and have a significant analgesic effect.

[0055] Test Example 2: Xylene-induced mouse ear swelling test Healthy male Kunming mice weighing 20-24g were randomly divided into 7 groups of 10 each. The blank group was coated with 1mL of physiological saline per mouse. The experimental groups were coated with 1mL of the drug composition for treating burns in intensive care prepared in Examples 1-3 or Comparative Examples 4-6 per mouse. The drugs were administered once a day for 5 consecutive days. 60 minutes after the last administration, under ether anesthesia, p-xylene was lightly applied to both sides of the left auricle of each mouse to induce inflammation. The right ear was left untreated as a normal control. 4 hours later, the mice were anesthetized and euthanized by cervical dislocation. The left and right ears were cut off along the auricle baseline, and circular ear pieces were punched at the same location on both ears. The ear pieces were weighed using an electro-optical analytical balance. The difference in weight between the left and right ear pieces of each mouse was used as the degree of auricle swelling. The average swelling degree was calculated, and the swelling inhibition rate was calculated based on the average swelling degree. The results are shown in Table 2.

[0056] Swelling degree = Mass of left ear flap with medication - Mass of right ear flap without medication Swelling inhibition rate = (Average swelling degree of the blank saline group - Average swelling degree of the drug-treated group) / Average swelling degree of the blank saline group × 100% Table 2

[0057] As can be seen from the table above, the pharmaceutical compositions for treating burns in intensive care prepared in Examples 1-3 of this invention have good anti-inflammatory effects.

[0058] Test Example 3 Healthy adult SD rats, weighing 200-240g, with half males and half females, were randomly divided into a blank group, a Jingwanhong group, Example 1-3 groups, and Comparative Example 4-6 groups, with 10 rats in each group. The rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (350mg / kg), and then the back of the rats was treated with 10% barium sulfide for hair removal. A 4cm diameter iron rod was placed in 100℃ hot water for 15min, and after being removed, the residual moisture on the surface was quickly absorbed with sterile filter paper. One end was then placed firmly on the treated skin on the back of the rat for 20s to establish a burn model.

[0059] The control group was coated with physiological saline, 2 mL / animal; the Jingwanhong group was coated with Jingwanhong ointment, 2 g / animal; the experimental group was coated with the corresponding prepared intensive care unit drug composition for treating burns, 2 mL / animal. Dressings were changed once daily after disinfection. Drug administration began 2 days after the burn and continued for 7 days. The wound area 2 days after the burn was used as the initial burn area. At the end of the experiment, transparent graph paper was laid over the burn site, and the wound area was traced; this was the final burn area. The traced graph paper was cut out and weighed; the wound area was calculated by using mass instead of area.

[0060] Wound healing rate (%) = (Initial burn area - Final burn area) / Initial burn area × 100% The results are shown in Table 3.

[0061] Table 3

[0062] Note: * indicates P < 0.05 compared to the control group.

[0063] As can be seen from the table above, the pharmaceutical compositions for treating burns in intensive care prepared in Examples 1-3 of this invention have a good effect on promoting wound healing after burns.

[0064] 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. A pharmaceutical composition for treating burns in intensive care, characterized in that, The raw materials include the following parts by weight: 1-1.5 parts of analgesic temperature-responsive liposomes, 0.5-1 part of co-loaded exosomes, 10-15 parts of Lithospermum erythrorhizon, 5-10 parts of Astragalus membranaceus, 3-7 parts of Rheum palmatum, 3-5 parts of Angelica dahurica, 3-5 parts of Angelica sinensis, 5-10 parts of Panax notoginseng, 10-15 parts of chitosan, and 20-25 parts of sodium β-glycerophosphate.

2. The pharmaceutical composition for treating burns in intensive care according to claim 1, characterized in that, Preparation of the analgesic temperature-responsive liposomes: S1. Ketamine and sufentanil were dissolved in hot ethanol, cooled to room temperature, and evaporated in an open container to obtain a eutectic. The eutectic was then filtered, washed, dried, and pulverized to obtain the ketamine and sufentanil eutectic. S2. Dipalmitoylphosphatidylcholine, cholesterol, ketamine and sufentanil cocrystal were added to diethyl ether, then added dropwise to buffer solution, stirred and mixed, the diethyl ether was removed by heating under reduced pressure, sonicated and filtered to obtain analgesic temperature-responsive liposomes.

3. The pharmaceutical composition for treating burns in intensive care according to claim 2, characterized in that, In step S1, the mass ratio of ketamine to sufentanil is 40-50:1, and the temperature of the hot ethanol is 60-70℃; in step S2, the mass ratio of dipalmitoylphosphatidylcholine, cholesterol, ketamine, and sufentanil cocrystal is 5-8:1-3:0.8-1.2, the buffer solution is PBS buffer with pH=7.4, the sonication time is 15-30 min, and the filtration is sequential filtration through 0.45μm and 0.22μm organic membranes.

4. The pharmaceutical composition for treating burns in intensive care according to claim 1, characterized in that, The method for preparing the co-loaded exosomes is as follows: T1. Centrifuge to remove exosomes from fetal bovine serum, prepare a culture medium for culturing deer antler stem cells, collect the supernatant after the cells have grown to confluence, and collect the deer antler stem cell exosomes by gradient centrifugation; T2. Dissolve salvianolic acid B in buffer solution, add deer antler stem cell exosomes, incubate, dialyze, and obtain the first drug-loaded exosomes; T3. Tanshinone IIA was dissolved in DMSO, mixed with an aqueous solution of the first drug-loaded exosomes, placed under an electroporator, electrolyzed, removed, centrifuged, and freeze-dried to obtain co-loaded exosomes.

5. The pharmaceutical composition for treating burns in intensive care according to claim 4, characterized in that, The mass ratio of salvianolic acid B, tanshinone IIA, and deer antler stem cell exosomes is 1-2:0.5-1:15-20; the culture medium in step T1 is DMEM complete culture medium containing 8-12% fetal bovine serum, 0.5-1.5wt% penicillin, and 0.5-1.5wt% streptomycin, and the deer antler stem cells are passage 5-7; the incubation temperature in step T2 is 36-38℃, the incubation time is 1-3h, the buffer is PBS buffer with pH=7.4, and the protein content of the first drug-loaded exosome aqueous solution is 0.3-0.4mg / mL; the number of electroshocks in step T3 is 3-7 times, and the voltage is 0.5-1V.

6. A method for preparing a pharmaceutical composition for treating burns in intensive care as described in any one of claims 1-5, characterized in that, Includes the following steps: (1) Mix Lithospermum erythrorhizon, Astragalus membranaceus, Rheum palmatum, Angelica dahurica, Angelica sinensis and Panax notoginseng, pulverize and sieve to obtain Chinese medicine powder. Add the Chinese medicine powder to ethyl acetate, heat and reflux to extract, filter, add emulsifier F68 and 1,2-propanediol to the filtrate to form an emulsion, add sodium alginate solution dropwise to emulsify, add petroleum ether to form a double emulsion, add calcium chloride solution dropwise to crosslink, centrifuge, wash and dry to obtain Chinese medicine microspheres; (2) Dissolve chitosan in acid to obtain chitosan solution, thus obtaining solution A; (3) Dissolve sodium β-glycerophosphate in sodium bicarbonate solution to obtain solution B; (4) Add solution B to solution A, add traditional Chinese medicine microspheres, analgesic temperature-responsive liposomes and co-loaded exosomes, stir and mix evenly to obtain a drug composition for treating burns in intensive care.

7. The preparation method according to claim 6, characterized in that, The heating and reflux extraction time in step (1) is 4-8h, the solid-liquid ratio of the Chinese herbal powder and ethyl acetate is 1:5-10g / mL, the concentration of the sodium alginate solution is 2-4wt%, the concentration of the calcium chloride solution is 1-3wt%, and the cross-linking time is 4-6h.

8. The preparation method according to claim 6, characterized in that, The acid solution mentioned in step (2) is a 1-3 wt% hydrochloric acid or acetic acid solution, and the concentration of the chitosan solution is 3-5 wt%.

9. The preparation method according to claim 6, characterized in that, In step (3), the concentration of sodium β-glycerophosphate in solution B is 50-60 wt%, and the concentration of sodium bicarbonate solution is 0.3-0.5 mol / L.

10. The preparation method according to claim 6, characterized in that, The drug composition for treating burns in the intensive care unit described in step (4) is liquid at room temperature and forms a gel at 37°C after being applied to the skin.

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