Application of picroside Ⅱ derivative in preparation of medicine for treating diabetic foot

Abstract

This invention provides the application of berberine II derivatives in the preparation of drugs for treating diabetic foot, belonging to the field of biomedical technology. This invention is the first to discover that berberine II derivatives can significantly improve symptoms such as decreased motor / sensory nerve conduction velocity and loss of pain sensation in diabetic foot model rats, and promote wound healing in the affected foot. Berberine II derivatives hold promise for further development and research as a novel drug for treating diabetic foot.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the application of berberine II derivatives in the preparation of drugs for treating diabetic foot. Background Technology

[0002] Diabetic foot (DF) is a disease caused by the combined effects of lower limb neuropathy, vascular disease, and infection in diabetic patients, resulting in tissue destruction and functional impairment of the foot. It is one of the most serious chronic complications of diabetes. Clinically, diabetic foot often manifests as abnormal foot sensation (such as numbness, dullness, and tingling), changes in skin temperature, and foot deformities (such as claw toes). As the disease progresses, foot ulcers and gangrene may occur, and in severe cases, amputation may be necessary. Among these factors, neuropathy leads to decreased foot sensation (such as not being aware of burns or abrasions), while vascular disease results in insufficient blood supply to the foot and weakened wound healing ability. The combination of these two factors easily induces infection, and the infection progresses rapidly, which is the core contributing factor to amputation.

[0003] According to data from the International Diabetes Federation (IDF), approximately 15% of diabetic patients worldwide develop diabetic foot ulcers, of which 20%-30% eventually require amputation. In my country, the amputation rate for diabetic foot patients is approximately 19.03%, and the 5-year survival rate after amputation is less than 50%, placing a heavy burden on patients and society.

[0004] The main pathological mechanisms of diabetic foot include: peripheral nerve axonal atrophy and demyelination, leading to sensory-motor nerve dysfunction; lower extremity arteriosclerosis, luminal stenosis, or even occlusion, causing foot ischemia and hypoxia; and the hyperglycemic environment damaging the foot's skin barrier, increasing susceptibility to infection. Current clinical treatment focuses on controlling blood sugar, improving circulation, fighting infection, and debridement and dressing changes. However, most measures only delay the progression of the disease and cannot reverse the pathological damage to nerves and blood vessels. Therefore, developing novel interventions for diabetic foot that can repair neurovascular damage and reduce the risk of amputation has significant clinical value and is an urgent practical need. Patent CN113773356A discloses the use of triazole-modified berberine II derivatives and studies their application in anticancer drugs. Based on this, this invention further explores the application prospects of berberine II derivatives in the treatment of diabetic foot, providing new drug options for the treatment of diabetic foot by broadening the application scope of berberine II derivatives. Summary of the Invention

[0005] (a) Technical problems to be solved

[0006] To address the shortcomings of existing technologies, this invention provides the application of berberine II derivatives in the preparation of drugs for treating diabetic foot.

[0007] (II) Technical Solution

[0008] To achieve the above objectives, the present invention provides the application of berberine II derivatives in the preparation of drugs for treating diabetic foot.

[0009] Specifically, the berberine II derivative has the following structure: Formula I

[0010]

[0011] Wherein, R is selected from one of phenyl, 2,5-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 2-chlorophenyl, 2,5-dichlorophenyl, 2,4,5-trichlorophenyl, 2-iodophenyl, 4-nitrophenyl, and piperidinylethyl.

[0012] Furthermore, the berberine II derivative is selected from any one of the following structures:

[0013]

[0014] Specifically, the drug for treating diabetic foot is made with a berberine II derivative of formula I or a pharmaceutically acceptable salt thereof as the active ingredient, supplemented with pharmaceutically acceptable excipients.

[0015] (III) Beneficial Effects

[0016] This invention is the first to discover that berberine II derivatives can significantly improve symptoms such as decreased motor / sensory nerve conduction velocity and loss of pain sensation in diabetic foot model rats, and promote wound healing in the affected foot. Berberine II derivatives hold promise for further development and research as a novel drug for the treatment of diabetic foot. Attached Figure Description

[0017] Figure 1 Photos showing the healing of diseased foot wounds in rats from each group of diabetic foot model rats.

[0018] Figure 2 Results of wound healing area in diabetic foot model rats in each group; * P<0.05; ** P<0.01; *** P<0.001. Detailed Implementation

[0019] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, 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.

[0020] Example 1

[0021] The derivatives 1-9 of berberine II were synthesized with reference to the published patent CN113773356A.

[0022] Example 2

[0023] Effects of berberine II derivative-3 on motor nerve conduction velocity, pain response and wound healing in diabetic foot model rats.

[0024] 1. Experimental Principle

[0025] This experiment is based on the symptoms of slowed nerve conduction velocity, decreased sensation, and slowed wound healing in diabetic foot patients. The nerve conduction velocity, mechanical pain threshold, thermal pain response time, and wound healing rate were measured in diabetic foot model rats to evaluate the ameliorative effect of berberine derivative-3 on peripheral neuropathy at the individual behavioral level.

[0026] 2. Experimental Materials and Methods

[0027] 2.1 Animal grouping and administration

[0028] Establishment of a diabetic rat model: Six-week-old male SD rats were purchased from Shanghai Slack and acclimatized for one week. Afterward, they were intraperitoneally injected with STZ (70 mg / kg), and blood glucose was measured on days 4-7. Three weeks after successful model establishment, the STZ-treated rats were anesthetized with isoflurane using an anesthesia machine, disinfected with alcohol, and then a 5 mm punch was used to create a wound on the dorsum of the right foot of the rat. The wound area was photographed and recorded. One minute later, the rats were placed in cages and allowed to recover. DFU modeling was then completed. Rats were then grouped according to body weight and blood glucose levels into a control group (non-diabetic rats), a model group (DFU rats), and a treatment group (berberine derivative-3, 15 or 30 mg / kg, berberine II 20 mg / kg), with eight rats in each group. Treatment lasted for four weeks via gavage. Mechanical pain and thermal pain sensitivity were measured weekly before and after drug administration. Motor nerve conduction velocity was measured before and after drug administration. Wound area and condition were photographed and recorded on days 3, 7, 14, 21, and 28.

[0029] 2.2 Mechanical Pain Threshold Experiment

[0030] This experiment is based on the principle of the retraction reflex in rodents when their claws are mechanically stimulated. The Vonfrey tactile measurement kit was purchased from Ugo Basile. Vonfrey fibers can provide a stimulating force ranging from 0.008 g to 300 g, with the fiber thickness determining the magnitude of the force. The experiment used fibers of appropriate thickness selected according to the actual situation, stimulating the skin vertically. The stimulation force was adjusted by changing the fibers until the fibers bent, causing the rats to exhibit a paw withdrawal response. This was used to assess the rats' response to pain. Each rat was measured six times to determine the threshold. The 50% mechanical pain threshold of the rat was calculated using the formula: 50% threshold = (10^(xf+kδ)) / 10000.

[0031] 2.3 Thermal pain latency test

[0032] The infrared plantar pain meter was purchased from Ugo Basile. A portable heat source was placed directly on the sole of the rat's hind paw for thermal stimulation. When the rat lifted its paw or moved away from the heat source, the instrument automatically stopped radiating and recorded the duration. The duration of this thermal latency was used to assess the rat's sensitivity to thermal pain.

[0033] 2.4 Motor Nerve Conduction Velocity Detection Experiment

[0034] ① Electrode Placement: The stimulating electrode is placed on the nerve trunk, the recording electrode on the muscle belly, and the reference electrode on the tendon; the ground wire is placed between the stimulating and recording electrodes. ② Calculation of Motor Nerve Conduction Velocity: Strong stimulation of the distal and proximal ends of the nerve trunk yields two compound muscle action potentials recorded on the muscles innervated by that nerve. The different latencies are measured, and the nerve conduction velocity is calculated by dividing the distance between the distal and proximal ends by the difference in latency between the two points. The formula is: Nerve conduction velocity (m / s) = Distance between two points (cm) × 10 / Latency difference between two points (ms).

[0035] 2.5 Sensory Nerve Conduction Velocity Test

[0036] ① Electrode Placement: The stimulating electrode is placed on the muscle belly, the recording electrode on the nerve trunk, and the reference electrode on the tendon; the ground wire is placed between the stimulating and recording electrodes. ② Calculation of Sensory Nerve Conduction Velocity: Strong stimulation of the muscle belly allows for the recording of action potentials on the nerve trunk responsible for sensation. Different latencies are measured, and the nerve conduction velocity is calculated by dividing the distance between the distal and proximal points by the difference in latency between the two points. The formula is: Nerve conduction velocity (m / s) = Distance between two points (cm) × 10 / Latency difference between two points (ms).

[0037] 2.6 Statistical Analysis

[0038] All data in this experiment were analyzed using SPSS 26.0 statistical software, and GraphPad Prism 8.0 software was used to generate statistical graphs. Experimental data are expressed as mean ± standard deviation, with a sample size of n=8 (8 rats per group), meeting the requirements of normal distribution and homogeneity of variance test. One-way ANOVA was used for comparisons among multiple groups, and LSD-t test was used for pairwise comparisons between groups. Specifically, quantitative data such as wound healing area of ​​the rat's diseased foot, motor nerve conduction velocity, mechanical pain threshold, and thermal pain response time were analyzed using the above methods. A p-value < 0.05 was considered statistically significant, and a p-value < 0.01 was considered extremely statistically significant, thus clarifying whether the effects of berberine derivative II-3 on relevant indicators in diabetic foot model rats were statistically significant.

[0039] 3. Experimental Results

[0040] 3.1 Effects of berberine II derivative-3 on pain response in diabetic foot rats

[0041] Tables 1 and 2 show the mechanical pain threshold and thermal pain latency of rats in each group. Before administration, compared with the control group, the mechanical pain threshold and thermal pain latency of diabetic foot model rats in the model group were significantly increased (P<0.001, P<0.001). After 4 weeks of administration, compared with the model group, the mechanical pain threshold and thermal pain latency of rats in each administration group were significantly decreased (P<0.05, P<0.01, P<0.001); among them, the decrease in mechanical pain threshold and thermal pain latency was more significant with the increase of the dose of berberine II derivative-3.

[0042] Table 1. 50% mechanical pain threshold of rats in each group (Mean ± SEM., n=8, g)

[0043]

[0044] Note: *p<0.05, **p<0.01, ***p<0.001 represent the model group vs. the control group; # p<0.05, ## p<0.01, ### p<0.001 represents the treatment group vs. the model group; the same applies below.

[0045] Table 2 Thermal pain latency in rats of each group (Mean ± SEM., n=8, s)

[0046]

[0047] 3.2 Effects of berberine II derivative-3 on the conduction velocity of motor and sensory nerves in diabetic foot rats

[0048] Tables 3 and 4 show the results of motor and sensory nerve conduction velocities in each group of rats. Before administration, compared with the control group, the motor and sensory nerve conduction velocities in the model group rats were significantly decreased (P<0.001, P<0.001). After 4 weeks of administration, compared with the model group, the motor and sensory nerve conduction velocities in each administration group rats were significantly increased (P<0.01, P<0.001); among them, the increase in motor and sensory nerve conduction velocities in diabetic foot rats was more significant with the increase of berberine derivative II-3 dose.

[0049] Table 3 Motor nerve conduction velocity of rats in each group (Mean±SEM., n=8, m / s)

[0050]

[0051] Table 4 Sensory nerve conduction velocities in rats of each group (Mean±SEM., n=8, m / s)

[0052]

[0053] 3.3 Effects of berberine II derivative-3 on wound healing in diabetic foot rats

[0054] Figure 1 and Figure 2 This study investigated the effect of berberine II derivative-3 on wound healing in diabetic foot rats. Compared to the control group, the model group rats showed slower wound healing, and the wound area in the model group remained consistently larger than that in the control group throughout the experiment. During the experiment, the wound area in the affected foot of rats in each treatment group was significantly smaller than that in the model group. At the end of the experiment, obvious wounds were still visible in the affected foot of rats in the model group, while the wounds in the affected foot of rats in each treatment group were essentially healed and invisible, comparable to those in the control group.

[0055] 4. Conclusion

[0056] The above experimental results indicate that berberine II derivatives can significantly improve symptoms such as decreased motor nerve conduction velocity and loss of pain sensation in diabetic foot model rats, and promote wound healing in the affected foot. Berberine II derivatives hold promise for further development and research as a novel drug for the treatment of diabetic foot.

[0057] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of the berberine II derivative of Formula I in the preparation of a drug for treating diabetic foot, wherein the structure of the berberine II derivative is as follows: ， in, The R is selected from one of phenyl, 2,5-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 2-chlorophenyl, 2,5-dichlorophenyl, 2,4,5-trichlorophenyl, 2-iodophenyl, 4-nitrophenyl, and piperidinylethyl.

2. The use of the berberine II derivative of formula I as described in claim 1 in the preparation of a drug for treating diabetic foot, characterized in that, The berberine II derivative is selected from any one of the following structures: 。 3. The use of the berberine II derivative of formula I as described in claim 1 in the preparation of a drug for treating diabetic foot, characterized in that, The drug for treating diabetic foot is formulated with a berberine II derivative of formula I or a pharmaceutically acceptable salt thereof as the active ingredient, supplemented with pharmaceutically acceptable excipients.

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