Application of rosmarinic acid in preparation of medicine for treating diabetic ulcer
By using rosemary acid as the only active ingredient topical drug, it promotes wound healing and re-epithelialization of diabetic ulcers and reduces ROS expression, the problem of poor treatment effect of diabetic ulcers in the prior art has been solved, and a significant effect of diabetic ulcer treatment has been achieved.
Patent Information
- Application Number
- CN202510153530.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-12
- Publication Date
- 2025-05-30
AI Technical Summary
The prior art is difficult to effectively treat diabetic ulcers, and the healing rate of diabetic ulcers is low and the recurrence rate is high, and there is a lack of efficient and simple treatment methods.
Rosemary acid is used as the only active ingredient to treat diabetic ulcer by promoting the healing and re-epithelialization of diabetic ulcers and reducing the expression of ROS in wounds.
Rosmarinic acid significantly promotes the healing of diabetic ulcers, reduces ROS expression, and improves the therapeutic effect of diabetic ulcers, providing a basis for the clinical development of efficient strategies for the treatment of diabetic ulcers.
Smart Images
Figure CN120053414A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of medicines, in particular to application of rosmarinic acid in preparing medicines for treating diabetic ulcers. Background Art
[0002] Diabetic foot ulcer is a serious complication of diabetic patients, characterized by foot infection and deep tissue damage caused by vascular and nerve abnormalities. The incidence of diabetic foot ulcers in diabetic patients is 19% to 34%, of which about 20% of cases eventually lead to amputation of varying degrees due to infection or other reasons. Diabetic foot ulcers are also one of the main causes of non-traumatic amputation of the lower limbs. The treatment of diabetic ulcers requires multidisciplinary collaboration. On the basis of adjusting blood sugar, blood lipids, and blood pressure, the treatment should follow the following principles: anti-infection, timely revascularization, decompression to reduce trauma to the ulcer site, and wound treatment to promote healing. However, only about 30%-40% of diabetic foot ulcers can heal within 12 weeks. Even if they heal, the recurrence rate is also very high, estimated to be 42% at 1 year and 65% at 5 years. Therefore, finding an efficient and simple treatment method for diabetic ulcer is a key issue that needs to be urgently solved in clinical practice (Wang K, Wang Y, Shi W, et al. Diagnosis and treatment of diabetic foot ulcer complicated with lower extremityvasculopathy: Consensus recommendation from the Chinese Medical Association(CMA), Chinese Medical Doctor Association (CMDA). Diabetes Metab Res Rev.2024 Mar;40(3):e3776. FIFE CE, ECKERT KA, CARTER M J. Publicly Reported Wound Healing Rates: The Fantasy and the Reality [J]. Advances in wound care,2018, 7(3): 77-94. ARMSTRONG DG, BOULTON AJM, BUS S A. Diabetic Foot Ulcers and Their Recurrence [J]. The New England journal of medicine, 2017,376(24): 2367-75.).
[0003] Rosmarinic acid (RA) is a natural polyphenolic compound, first isolated from rosemary, and widely present in various Chinese medicinal herbs such as perilla leaves, salvia miltiorrhiza, scutellaria baicalensis, motherwort, mint, and honeysuckle. It is convenient to extract and has a relatively low price. Rosmarinic acid has a variety of biological activities and pharmacological effects, including antioxidant, anti-inflammatory, antibacterial, antiviral, anticancer, and immunomodulatory effects (Sirajudeen F, Bou Malhab LJ, Bustanji Y, et al. Exploring the Potential of Rosemary Derived Compounds (Rosmarinic and Carnosic Acids) as Cancer Therapeutics: Current Knowledge and Future Perspectives. Biomol Ther (Seoul). 2024 Jan 1;32(1):38 - 55.).
[0004] At present, there is no relevant report in this field on the correlation between rosmarinic acid and the treatment of diabetic ulcers, and the development of new drugs for the treatment of diabetic ulcers is of great significance. Summary of the Invention
[0005] The purpose of the present invention is to provide a new use of rosmarinic acid in the treatment of diabetic ulcers, and rosmarinic acid has a significant curative effect in the treatment of diabetic ulcers.
[0006] In the first aspect of the present invention, there is provided the use of rosmarinic acid in the preparation of a drug for the treatment of diabetic ulcers.
[0007] Furthermore, the molecular formula of the rosmarinic acid is C 18 H 16 O 8 , and its chemical structural formula is shown as Formula I below:
[0008]
[0009] Formula I
[0010] The present invention has no special limitation on the specific source of rosmarinic acid, which can be obtained by self - extraction and preparation, or purchased commercially.
[0011] Furthermore, rosmarinic acid exerts its effect by promoting the healing and re - epithelialization of diabetic ulcer wounds and reducing the expression of ROS in diabetic ulcer wounds.
[0012] Furthermore, in the above - mentioned use, there is provided the use of rosmarinic acid in the preparation of a drug for promoting the healing and re - epithelialization of diabetic ulcer wounds.
[0013] Further, in the above application, rosmarinic acid is used in the preparation of a drug for reducing the expression of ROS in diabetic ulcer wounds.
[0014] In the second aspect of the present invention, a drug for treating diabetic ulcers is provided, and the drug uses rosmarinic acid as the sole active ingredient.
[0015] Further, the drug is an external preparation. More specifically, the drug further includes excipients commonly used in external preparations.
[0016] The present invention has no special limitation on the types and specific sources of excipients commonly used in external preparations, and common excipients in the field of external preparations can be used.
[0017] Further, the effective dose of rosmarinic acid in the drug in vivo is 20 mM.
[0018] The advantages of the present invention are as follows:
[0019] 1. The present invention discovers for the first time that the traditional Chinese medicine monomer rosmarinic acid has the effect of treating diabetic ulcers and has achieved remarkable therapeutic effects, providing a basis for the future clinical development of efficient strategies for treating diabetic ulcers.
[0020] 2. The present invention uses a mouse diabetic ulcer model to study the effects of rosmarinic acid on diabetic ulcer healing indicators and ROS expression. The results show that rosmarinic acid has a significant therapeutic effect on diabetic ulcers, providing important experimental evidence for the future clinical development of efficient strategies for treating diabetic ulcers and the preparation of drugs for treating diabetic ulcers. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 Shows the effect of rosmarinic acid on the wound healing of diabetic ulcer mice. Among them, Figure A shows the results of wound healing photography and the calculation results of wound healing rate; Figure B shows the results of HE staining and the quantitative calculation results of HE, scale bar = 1 mm.
[0022] Figure 2 Shows the effect of rosmarinic acid on the ROS expression in the wounds of diabetic ulcer mice. Among them, the left figure shows the results of fluorescence microscope detection of ROS in the wound surface, and the right figure shows the quantitative calculation results of fluorescence microscope detection of ROS in the wound surface. DETAILED DESCRIPTION OF THE INVENTION
[0023] The following detailed description of the specific embodiments provided by the present invention will be given in conjunction with the examples and the drawings. In the following examples, unless otherwise specified, all are conventional methods. The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.
[0024] Example 1: Effect of rosmarinic acid on wound healing in diabetic ulcer mice
[0025] 1. Animals: m / m control mice and db / db diabetic mice were used in the animal experiment. They were male, 6 - 8 weeks old, provided by the Experimental Animal Center of Yueyang Hospital of Integrated Traditional Chinese and Western Medicine Affiliated to Shanghai University of Traditional Chinese Medicine, and purchased from GemPharmatech Co., Ltd. All animals were housed in an environment with a light - dark cycle, the temperature was maintained at 23 ± 2 °C, and they had free access to water and food.
[0026] Animal model: Five m / m mice were used as the normal control group, and 15 db / db diabetic mice were randomly and evenly divided into a diabetic group, a diabetic rosmarinic acid treatment group, and a diabetic positive treatment group. The hair on the backs of m / m normal control and db / db diabetic mice was shaved, and the shaved area was about 4×4 cm. Before making the ulcer model, the mice were anesthetized with isoflurane, then the back skin was pinched up, and a wound model with an area of 6×6 mm was made using a punch. Each mouse had 4 wounds on the back, and the experiment was carried out under sterile conditions.
[0027] 2. Grouping and treatment:
[0028] (1) Normal group (m / m): The ulcer model was made, and it was treated with the drug base solvent (10% DMSO, 40% PEG300, 5% Tween - 80, 45% Saline). It was wet - applied to the ulcer of the mice at a dose of 80 μl / rat / day for 9 consecutive days; (2) Diabetic group (db / db): The ulcer model was made, and it was treated with the drug base solvent, wet - applied to the ulcer of the mice at a dose of 80 μl / rat / day for 9 consecutive days; (3) Diabetic rosmarinic acid treatment group (db / db + RA): Rosmarinic acid monomer was dissolved in the base solvent (10% DMSO, 40% PEG300, 5% Tween - 80, 45% Saline) at a concentration of 20 mM, and it was wet - applied to the ulcer of the mice at a dose of 80 μl / rat / day for 9 consecutive days; (4) Diabetic positive treatment group (db / db + rb - bFGF): The bovine basic fibroblast growth factor topical solution, with a specification of 63000 IU / bottle, was topically applied to the ulcer of the mice at a dose of 80 μl / rat / day for 9 consecutive days.
[0029] 3. Experimental methods:
[0030] (1) Wound healing analysis
[0031] Macroscopic observation: Photos were taken on the 1st, 3rd, 5th, 7th, and 9th days after drug administration and treatment. The Image - J software was used to process the photographed pictures to calculate the ulcer area of the wound surface. The following formula was used to calculate the wound closure:
[0032] Wound healing rate (%) = (1 - Treated wound width / Control wound width) × 100
[0033] (2) Histological hematoxylin-eosin staining (HE staining)
[0034] On the 9th day after induction, the mice were sacrificed, and the skin specimens were fixed in 4% neutral formaldehyde buffer for 24 h. Routine histological examination was performed, including paraffin embedding, sectioning, and HE staining. The sections were observed under a digital slide scanner.
[0035] 4. Experimental results
[0036] Rosmarinic acid can improve wound healing in diabetic ulcer mice.
[0037] Wound healing analysis and HE staining were used to observe the effect of rosmarinic acid on diabetic ulcer wound healing in vivo. The results are as Figure 1 shown. As can be seen from Figure 1 A, compared with the diabetic positive treatment group, the diabetic rosmarinic acid treatment group had more significant wound healing effects on the 5th, 7th, and 9th days after injury. The HE staining results ( Figure 1 B) showed that the wound in the diabetic rosmarinic acid treatment group was significantly reduced on the 9th day, and the effect was better than that of the diabetic positive treatment group. These results indicate that the wound healing of diabetic mice is slowed down, and rosmarinic acid intervention can effectively promote diabetic wound healing and re-epithelialization, and the curative effect is more significant than that of the positive control group.
[0038] Example 2: Effect of rosmarinic acid on the expression of ROS in the wound surface of diabetic ulcer mice
[0039] 1. Animal models: m / m control mice and db / db diabetic mice.
[0040] 2. Grouping and treatment:
[0041] (1) Normal group (m / m): Ulcer models were established and treated with the drug base solvent (10% DMSO, 40% PEG300, 5% Tween-80, 45% Saline). The ulcers of mice were wet compress with a dosage of 80 μl / rat / day for 9 consecutive days. (2) Diabetic group (db / db): Ulcer models were established and treated with the drug base solvent. The ulcers of mice were wet compress with a dosage of 80 μl / rat / day for 9 consecutive days. (3) Diabetic rosmarinic acid treatment group (db / db+RA): Rosmarinic acid monomer was dissolved in the base solvent at a concentration of 20 mM. The ulcers of mice were wet compress with a dosage of 80 μl / rat / day for 9 consecutive days. (4) Diabetic positive treatment group (db / db+rb-bFGF): The bovine basic fibroblast growth factor topical solution with a specification of 63000 IU / bottle was topically applied to the ulcers of mice at a dosage of 80 μl / rat / day for 9 consecutive days.
[0042] 3. Experimental methods:
[0043] (1) ROS fluorescence staining and semi-quantitative analysis in the wound surface
[0044] On the 9th day after induction, the mice were sacrificed, and the skin specimens of the wound surface were taken. The ROS expression in the wound surface was detected using the dihydroethidium ROS detection kit. Add 200 μl of washing solution to 10 µm frozen sections for 5 min, remove the washing solution, add 200 μl of 60 μM dihydroethidium fluorescence probe diluted with PBS for 60 min, remove the dihydroethidium fluorescence probe, and wash 3 times with PBS. Detect ROS under a fluorescence microscope. Use Image-J software to perform semi-quantitative analysis of the average fluorescence intensity of the fluorescence images. The average fluorescence intensity was calculated using the following formula:
[0045] Average fluorescence intensity = sum of fluorescence intensity in the area / area of the area
[0046] 4. Experimental results:
[0047] Rosmarinic acid can reduce the ROS expression in the wound surface of diabetic ulcer mice.
[0048] The ROS content was detected using the dihydroethidium ROS detection kit to observe the effect of rosmarinic acid on the ROS expression in the wound surface of diabetic ulcer mice. The results are as Figure 2 shown. Compared with the diabetic positive treatment group, the ROS content in the wound surface of the diabetic rosmarinic acid treatment group was significantly lower than that of the diabetic positive treatment group. This result indicates that the ROS expression in the wound surface of db / db diabetic mice is abnormally increased, and rosmarinic acid intervention can effectively reduce the ROS expression in diabetic wounds, and the curative effect is more significant than that of the positive control group.
[0049] The preferred embodiments of the present invention have been specifically described above. However, the present invention is not limited to the described embodiments. Those skilled in the art can still make various equivalent modifications or substitutions without departing from the spirit of the present invention. These equivalent modifications or substitutions are all included within the scope defined by the claims of this application.
Claims
1. Application of rosmarinic acid in the preparation of medicines for treating diabetic ulcers.
2. The use of rosmarinic acid according to claim 1 in preparing a medicament for treating diabetic ulcer, characterized in that: Application of rosmarinic acid in the preparation of drugs for promoting wound healing and re-epithelialization of diabetic ulcers.
3. The use of rosmarinic acid according to claim 1 in preparing a drug for treating diabetic ulcers, characterized in that: Application of rosmarinic acid in the preparation of drugs for reducing ROS expression in diabetic ulcer wounds.
4. A drug for treating diabetic ulcer, characterized in that: The drug has rosmarinic acid as the only active ingredient.
5. The drug according to claim 4, characterized in that The medicine is an external preparation.
6. The drug according to claim 5, characterized in that The medicine also includes auxiliary materials commonly used in external preparations.
7. The drug according to claim 4, characterized in that The effective dose of rosmarinic acid in the medicine in vivo is 20 mM.