Use of breviscapine-RGD peptide compound in preparation of drugs for treating circulatory disturbance and inflammation of extremities caused by frostbite

Abstract

The application discloses application of a scutellarein-RGD peptide compound in preparation of drugs for treating circulatory disturbance and inflammation of extremities caused by frostbite. The application discloses a new application of the scutellarein-RGD peptide compound in the field of treating frostbite. The scutellarein-RGD peptide compound effectively improves microvascular thrombosis, microcirculation, blood activation and recovery. The scutellarein-RGD peptide compound has obvious improvement effect on ischemia-reperfusion injury, inflammation and edema of lower limbs caused by frostbite, and obviously improves the recovery efficiency of extremities of frostbite legs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to the application of a scutellarin-RGD peptide compound in the preparation of a drug for treating circulatory disorders and inflammation in the extremities caused by frostbite. Background Technology

[0002] Frostbite is a systemic or localized freezing injury caused by exposure to low temperatures. It is more common in cold regions and often occurs in areas with poor peripheral circulation or exposed areas, such as the hands, feet, ears, nose, and cheeks. Frostbite of the lower limbs can cause circulatory disorders in the extremities, leading to ischemia and subsequent ischemia-reperfusion, which has adverse effects on the body.

[0003] Erigeron breviscapus extract has broad and definite pharmacological activities, and has a certain protective effect against liver, kidney and lung damage. It is widely used in the treatment of tissue ischemia, embolism and insufficient blood supply in organs such as heart, brain, lung, liver and kidney, and has achieved good clinical efficacy. It also has considerable efficacy in dilating cardiovascular and cerebrovascular vessels, anticoagulation, various thrombotic diseases, and reducing early kidney lesions in type II diabetes.

[0004] The scutellarin-RGD peptide compound is a compound obtained by structural modification of scutellarin B. Currently, there are no reports on the use of the scutellarin-RGD peptide compound for treating frostbite-induced peripheral circulatory disorders and inflammation. The chemical structure of the scutellarin-RGD peptide compound is as follows: Summary of the Invention

[0005] In order to overcome the above-mentioned shortcomings of the existing technology, the purpose of this invention is to provide the application of scutellarin-RGD peptide compounds in the preparation of drugs for treating frostbite-induced peripheral circulatory disorders and inflammation.

[0006] The technical solution of the present invention to solve the above-mentioned technical problems is as follows: Application of scutellarin-RGD peptide compounds in the preparation of drugs for treating circulatory disorders and inflammation in the extremities caused by frostbite.

[0007] Furthermore, the drug is a formulation prepared from an active ingredient and excipients, wherein the active ingredient includes scutellarin-RGD peptide compounds and pharmaceutically acceptable scutellarin-RGD peptide compound derivatives, and the excipients include pharmaceutically acceptable excipients.

[0008] Furthermore, drug dosage forms include topical preparations, oral preparations, or injectable preparations.

[0009] Furthermore, topical preparations include emulsions, creams, patches, sprays, or ointments.

[0010] Furthermore, emulsions, creams, patches, sprays, or ointments use scutellarin-RGD peptide compounds or pharmaceutically acceptable scutellarin-RGD peptide compound derivatives as active ingredients, and then add pharmaceutically acceptable dressings such as polyethylene glycol, glycerin, vegetable oils, fillers, and chitosan.

[0011] The present invention has the following beneficial effects: (1) Animal experiments have shown that after frostbite, the ligustrazine-RGD peptide compound can effectively improve microvascular thrombosis, microcirculation and blood circulation recovery.

[0012] (2) Animal experiments have shown that the present invention has a significant effect on improving lower limb ischemia-reperfusion injury, inflammation and edema caused by frostbite.

[0013] (3) Animal experiments have shown that the present invention has significantly improved the recovery efficiency of frostbitten legs by ligustrazine-RGD peptide compound. Attached Figure Description

[0014] Figure 1 The recovery of hind limb blood flow in each group of rats in Example 1 was shown at different time points. Figure 2 This refers to the recovery of limb swelling in each group of rats in Example 1; Figure 3 The appearance recovery of the hind limbs of rats in each group in Example 1; Figure 4 The SOD inhibition rate of the high-dose group and the model group in Example 2. Detailed Implementation The examples given below are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified, conditions in the examples are performed under standard conditions or as recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0015] Example 1: 1. Blood flow restoration experiment (1) Laboratory animals Twenty experimental-grade female SD rats were used for a week of acclimatization before the experiment was conducted.

[0016] (2) Grouping Twenty rats were divided into five groups according to their body weight: blank control group, model group, positive drug group, high-dose group, and low-dose group.

[0017] The ligustrazine-RGD peptide compound and aspirin were prepared into ligustrazine solutions with concentrations of 1.25 mg / ml and 0.5 mg / ml, and an aspirin aqueous solution with a concentration of 0.5 mg / ml, respectively, using 0.9% saline. These were administered intraperitoneally for seven consecutive days.

[0018] Positive drug group: Aspirin aqueous solution was administered at a dose of 10 mg / kg; High-dose group: The drug was administered with a 1.25 mg / ml solution of scutellarin, at a dose of 20 mg / kg; Low-dose group: The drug was administered using a 0.5 mg / ml solution of scutellarin, at a dose of 5 mg / kg.

[0019] (3) Modeling method Seven days after drug administration, modeling began. Two magnets with the same surface area were used to adhere to metal hemostats and pre-cooled in liquid nitrogen for at least 15 minutes to ensure their temperature matched the liquid nitrogen. The animals were then anesthetized with 2% sodium pentobarbital at a dose of 0.2 ml / 100 g. After anesthesia, the animals were placed in a prone position, and the skin of the right hind limb was incised to expose the tibialis anterior muscle. A pre-cooled circular magnet with a radius of approximately 0.8 cm was directly pressed onto the surface of the tibialis anterior muscle, with the two magnets being alternately frozen and replaced every 30 seconds. A preliminary experiment was conducted, and the treatment lasted 25 minutes (after freezing, the muscle tissue at the contact site became pale and hard, gradually turning from white to dark red after about seven or eight minutes). After rinsing with sterile irrigation solution, the skin was sutured directly. Following the modeling surgery, the animals' diet, activity, mental state, wound healing, and other general conditions were observed and recorded. Normal food and water were ensured.

[0020] (4) Test methods Throughout the modeling process, blood flow in the right hind limb was monitored using a blood flow meter to ensure consistent ischemia time and severity. Blood flow was recorded before modeling, at the end of modeling, 20 minutes after modeling, and 40 minutes after modeling. Cross-sectional comparisons were performed between groups.

[0021] (5) Test results The test results are as follows Figure 1 As shown, the experimental results indicate that the high-dose group and the positive control group showed significantly better blood flow recovery than the model group within the same time frame, while the low-dose group showed little difference from the model group. This demonstrates that a certain dose of scutellarin-RGD peptide compound significantly improves microvascular thrombosis, microcirculation, and blood circulation recovery in rats after frostbite.

[0022] 2. Hind limb swelling test (1) Test methods Forty-eight hours after modeling, the rats were anesthetized. Using a limb edema meter, a horizontal line was drawn at the knee joint of the hind limbs to ensure that the measurement range of both legs was consistent. The hind limbs were immersed in the measuring solution, with the liquid level flush with the marked line, and the volume of both hind limbs of the rats in each group was measured. The mean was calculated, and the lateral comparisons between the groups were performed.

[0023] (2) Test results The test results are as follows Figure 2 As shown in the results, the difference in volume between the frostbitten leg and the normal leg in the high-dose group was the smallest. The average value of the positive control group was slightly higher than that of the high-dose group, but still significantly lower than that of the low-dose group and the model group. There was no significant difference between the low-dose group and the model group. This indicates that the scutellarin-RGD peptide compound has a significant ameliorative effect on frostbite-induced lower limb ischemia-reperfusion injury, inflammation, and edema.

[0024] 3. Limb recovery test (1) Test methods Five days after the model was established, the rats were placed on a flat surface to walk, and it was observed whether the frostbitten hind limbs showed signs of curling, lameness, or impaired normal movement.

[0025] (2) Test results The test results are as follows Figure 3 As shown in the results, the high-dose group of rats showed better recovery of the frostbitten extremities, exhibiting slight lameness after 5 days, but still able to walk freely. The positive control group showed similar recovery to the high-dose group, but lameness persisted. The low-dose group and the model group showed poorer recovery, with significant limb curling and lameness, affecting their normal walking. This indicates that the ligustrazine-RGD peptide compound promotes the recovery of frostbitten extremities in rats.

[0026] Example 2: SOD content detection The experimental animals, grouping, and modeling methods are the same as in Example 1.

[0027] (1) Test methods Under normal circumstances, the body maintains a balance between oxidation and antioxidants. Peroxides and oxygen free radicals produced by oxidation are continuously eliminated by antioxidants in the body, thus preventing damage. However, in severe frostbite, this balance is disrupted, with increased oxidation and weakened antioxidant processes, leading to injury. Superoxide dismutase (SOD) protects against cellular immune damage by scavenging free radicals, resulting in a significant decrease in SOD activity 48 hours after frostbite, sometimes even below pre-frost levels. Therefore, detecting SOD levels in the heart after frostbite can provide a direct assessment of the individual's injury severity.

[0028] Rats were harvested 48 hours after modeling, and the SOD inhibition rate in each group was detected using a total SOD kit. (2) Test results The test results are as follows Figure 4 As shown in the figure, the highest and lowest SOD inhibition rates in the high-dose group and the model group are respectively. The results show that the total SOD inhibition rate in the heart homogenate of rats in the high-dose group is significantly higher than that in the model group. This indicates that the SOD content in the high-dose group rats was significantly higher than that in the model group 48 hours after modeling, while the SOD content in the model group rats was lower and the degree of oxidation was more severe. This suggests that the ligustrazine-RGD peptide compound can alleviate the effects of lower limb ischemia-reperfusion on distal organs.

[0029] 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. The application of scutellarin-RGD peptide complex in the preparation of drugs for treating frostbite-induced peripheral circulatory disorders and inflammation, characterized in that, The structural formula of the scutellarin-RGD peptide compound is as follows: 。 2. The application according to claim 1, characterized in that, The drug is a formulation prepared from an active ingredient and excipients, wherein the active ingredient includes scutellarin-RGD peptide compounds, and the excipients include pharmaceutically acceptable excipients.

3. The application according to claim 1, characterized in that, The drug dosage forms include topical preparations, oral preparations, or injectable preparations.

Images