Betulinol ester preparation for preventing and treating cognitive impairment after stroke and application of betulinol ester preparation
By using betulin preparations, the problem of preventing and treating post-stroke cognitive impairment has been solved. By regulating signaling pathways and inhibiting neuroinflammation, post-stroke cognitive impairment has been improved, achieving safe and effective treatment results.
Patent Information
- Application Number
- CN202511923779.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-11
- Publication Date
- 2026-02-13
AI Technical Summary
Existing drugs cannot effectively stop the progression of post-stroke cognitive impairment, and its pathogenesis is complex, lacking effective prevention and treatment methods.
Betulinum ether is used as the active pharmaceutical ingredient to prepare granules, powders, oral liquids and other dosage forms for the treatment and prevention of post-stroke cognitive impairment. It reduces neuronal oxidative stress by regulating signaling pathways, inhibits microglia activation, penetrates the blood-brain barrier, reduces ROS levels and improves neurological function.
It significantly improves post-stroke cognitive function, enhances cognitive and behavioral abilities, reduces neuronal loss, stabilizes synaptic connections, inhibits neuroinflammation, and provides a safe and effective treatment option.
Smart Images
Figure CN121513014A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to a new use of betulinic acid ester, in particular to the application of betulinic acid ester in preventing and treating post-stroke cognitive impairment, and belongs to the field of biological pharmacy. BACKGROUND
[0002] Post-stroke cognitive impairment (PSCI) is a common complication of ischemic stroke and a common neurological disease in the elderly, characterized by memory, visual spatial and executive function impairment, with a prevalence of 24%-53.4%, which seriously affects the quality of life and survival rate of patients after stroke, and brings a huge burden to the patient's family and society. The pathogenesis of post-stroke cognitive impairment is extremely complex, and the main mechanisms include inflammatory response, oxidative stress, mitochondrial dysfunction, neuronal damage, etc., but the exact pathogenesis is not clear. Recent studies have emphasized the close relationship between neuroinflammation and the occurrence and progression of post-stroke cognitive impairment, activated microglia exacerbate neuroinflammation by releasing cytotoxic mediators, including inflammatory cytokines and reactive oxygen species (ROS), which exacerbate the destruction of the blood-brain barrier (BBB) and aggravate secondary neuronal damage. At present, the clinical drugs for post-stroke cognitive impairment mainly improve brain blood circulation and strengthen brain nutrition, however, these drugs or measures cannot prevent the disease progression of post-stroke cognitive impairment. Therefore, it is imminent to screen effective drugs for preventing and treating post-stroke cognitive impairment.
[0003] Betulinic acid ester is a natural triterpenoid compound extracted from the bark of white birch, which has anti-inflammatory, antioxidant and neuroprotective biological activities. Studies have shown that betulinic acid ester can reduce oxidative stress in neurons by regulating various signaling pathways. In diabetic mouse models, betulinic acid ester can lower blood glucose levels and improve learning and memory abilities. Research has found that betulinic acid ester can inhibit ROS production, activate AMPK to promote the phosphorylation and nuclear translocation of transcription factor FoxO1, and then activate lysosomal biogenesis and autophagy processes. Betulinic acid ester can significantly reduce the levels of pro-inflammatory mediators TNF-alpha, matrix metalloproteinase (MMP-2), IL-1 beta, IL-2, IL-4, IL-5, IL-6, IL-13 and IL-17. Betulinic acid ester can improve the damage of neural function in animal models, inhibit the activation of microglial cells, and reduce the levels of pro-inflammatory factors TNF-alpha, IL-1 beta and IL-6 in the hippocampus, while increasing the level of anti-inflammatory factor IL-10. Betulinic acid ester can penetrate the blood-brain barrier (BBB), which can effectively reduce ROS levels and reduce neuronal apoptosis. Betulinic acid ester can effectively up-regulate antioxidant enzymes heme oxygenase 1 (HO-1) and superoxide dismutase 2 (SOD2) to reduce ROS levels, up-regulate the Nrf2 signaling pathway, and improve neuronal apoptosis and oxidative damage after subarachnoid hemorrhage. Betulinic acid ester can significantly reduce the burden of oxygen free radicals, reduce neuronal loss, and stabilize synaptic connections. In addition, betulinic acid ester can significantly reduce the expression level of keap1, while increasing the expression levels of HO-1 and Nrf2. Betulinic acid ester can activate the Nrf2 signaling pathway, promote the expression level of HO-1 and nuclear translocation of Nrf2. The antioxidant effect of betulinic acid ester may come from the activation of the Nrf2 signaling pathway and the reduction of free radical levels. Currently, betulinic acid ester has the effects of improving inflammatory response and inhibiting oxidative stress, but there is no report on the application of betulinic acid ester in the prevention and treatment of post-stroke cognitive impairment. SUMMARY
[0004] The purpose of the present application is to overcome the defects and deficiencies of the prior art, and to creatively apply betulinic acid ester in the prevention and treatment of post-stroke cognitive impairment, thereby developing a preparation for preventing and treating post-stroke cognitive impairment.
[0005] The purpose of the present application is to provide a safe and effective preparation for preventing and treating post-stroke cognitive impairment and its application.
[0006] Another purpose of the present application is to provide a new use of betulinic acid ester, i.e. the application of betulinic acid ester in the preparation of a composition for preventing and treating post-stroke cognitive impairment, which can be a drug, a health product and a beverage, etc.
[0007] The present application is to use betulinic acid ester as the active ingredient in an effective dose, and to use conventional excipients, nutrients and other auxiliary materials to prepare any kind of preparation for treating and preventing post-stroke cognitive impairment according to a certain preparation process. The dosage form used in clinic can be granules, powder, oral liquid, enema, etc.
[0008] The effective dose referred to in the present application means that the effective dose contained in the solid preparation prepared by using betulinic acid ester as the active ingredient according to the above-mentioned single or combination cannot be less than 1 g / kg.
[0009] Since the present application discloses for the first time the use of betulinic acid ester as the active ingredient in the preparation of a drug for treating and preventing post-stroke cognitive impairment, betulinic acid ester is combined with auxiliary materials to prepare a medicament.
[0010] Advantages and effects of the present application:
[0011] The present application discloses for the first time the use of betulinic acid ester in preventing and treating post-stroke cognitive impairment. The present application shows that betulinic acid ester can improve the cognitive impairment after stroke and improve the cognitive behavior. Therefore, the betulinic acid ester composition of the present application can be used as a drug for preventing and treating post-stroke cognitive impairment, has no toxic side effects, and can be used for the auxiliary treatment of patients with post-stroke cognitive impairment in clinic. BRIEF DESCRIPTION OF DRAWINGS
[0012] Figure 1 . Betulinic acid ester improves the cognitive impairment of post-stroke cognitive impairment mice; (A) nesting score in nesting test; (B) recognition index in object recognition test; (C) latency in Barnes maze test; (D) time spent in target quadrant in Barnes maze test; Nesting Score: nesting score in nesting test;
[0013] Recognition Index: recognition index in object recognition test; Latency: latency in Barnes maze test; Time speed in target quadrant: time spent in target quadrant in Barnes maze test; * : P<0.05; ** : P<0.01; WT: normal control group; PSCI: post-stroke cognitive impairment animal model group; PSCI+BE: betulinic acid ester treatment group for post-stroke cognitive impairment animal model.
[0014] Figure 2. Betulinol improved the pathological changes of PSCI mice after stroke; Representative immunofluorescence images of NeuN (green), scale bar 100 pm, magnification 200x; NeuN is a neuron-specific nuclear protein; Merge: the images collected in different fluorescence channels are superimposed to form a composite image; WT: normal control group; PSCI: animal model group of post-stroke cognitive impairment; PSCI+BE: betulinol treatment group for post-stroke cognitive impairment animal model;
[0015] Figure 3 . Betulinol inhibits neuroinflammation in PSCI. Representative immunofluorescence images of Iba1 (green), scale bar 50 pm, magnification 400x; Iba1, a polarization marker protein for microglia; DAPI (4', 6-diamidino-2-phenylindole) is a small molecule fluorescent dye, its core function is to specifically bind to the adenine and thymine-rich region in the DNA double helix structure; Merge, the images collected in different fluorescence channels are superimposed to form a composite image; WT: normal control group; PSCI: animal model group of post-stroke cognitive impairment; PSCI+BE: betulinol treatment group for post-stroke cognitive impairment animal model. DETAILED DESCRIPTION
[0016] The betulinol composition of the present application can be added with corresponding excipients as needed to form corresponding dosage forms.
[0017] Example 1 Animal experiment of betulinol for prevention and treatment of post-stroke cognitive impairment
[0018] Objective: To observe the neuroprotective effect of betulinol on post-stroke cognitive impairment model mice.
[0019] I. Materials and methods
[0020] 1. Experimental animals and treatment: unilateral common carotid artery occlusion was used to induce post-stroke cognitive impairment (PSCI) mouse model. Experimental animals were divided into three groups: WT group (normal control group), PSCI group (post-stroke cognitive impairment model group) and PSCI+BE group (betulinol treatment for post-stroke cognitive impairment group). Mice in the PSCI+BE group were treated with 50 mg / kg of betulinol daily for 3 weeks. Mice in the PSCI and Con groups were given an equal amount of normal saline. After treatment, behavioral tests such as nest building, novel object recognition and Barnes maze were performed to assess the cognitive function of mice.
[0021] 2. Test of cognitive function
[0022] A) Nest building test: The nest building test was performed in a clean cage, and 30 pieces of paper (5 cm x 5 cm) were evenly placed on the bedding. After 12 h, the nest building state of the mice was observed, and the results were recorded by photography.
[0023] B) Novel object recognition test: This test was performed in an open paper box (25 cm x 25 cm x 35 cm). In the training phase, each mouse was placed in a box containing two identical rectangular wooden blocks and allowed to explore freely for 5 min. After 24 h, one of the rectangular wooden blocks was replaced with a cylindrical wooden block. The time spent by each mouse exploring the novel object (TN) and the familiar object (TF) was recorded. After each test, the box was thoroughly cleaned with alcohol to eliminate any residual odors or traces that might interfere with subsequent tests. The recognition index (RI) was calculated using the formula: RI = TN / (TN + TF) x 100%.
[0024] C) Barnes maze test: This test was performed on a circular platform with a diameter of 100 cm, surrounded by 20 equally spaced escape holes, one of which was connected to a black escape box. The test protocol consisted of three phases: adaptation (day 1), training (days 2-5), and testing (day 6). In the adaptation phase, each mouse was gently guided into the target hole and allowed to adapt to the escape box for 4 min. In the training phase, each mouse was placed in a plastic cylinder (20 cm in diameter and 27 cm high) in the center of the maze and restricted from moving for 5 s. The cylinder was then removed, and the mouse was allowed to search for the escape box for 3 min. In the testing phase, the circular platform was divided into four quadrants, and the target box was removed. The time spent by the mouse in the target quadrant was recorded.
[0025] 3. Histopathology: After the behavioral tests, the brain tissue of the mice was fixed in 4% paraformaldehyde for 24 h, dehydrated in a graded ethanol series, and cleared with xylene. The samples were then embedded in wax blocks, cut into 5 pm paraffin sections, and baked in a incubator at 65°C for 2 h. The sections were deparaffinized with xylene and rehydrated through a graded ethanol series. The sections were subjected to antigen retrieval in citrate buffer for 10 min and blocked in 5% bovine serum albumin (BSA) and 0.5% Triton X-100 at room temperature for 1 h. The sections were incubated with the primary antibody overnight at 4°C, washed three times in phosphate-buffered saline, incubated with the secondary antibody in 5% BSA at room temperature for 1 h, and subjected to microscopic imaging using a fluorescence microscope at room temperature. Image analysis was performed using ImageJ software.
[0026] II. Experimental results:
[0027] 1. Betulinol improved cognitive impairment and pathological changes in mice with post-stroke cognitive impairment.
[0028] To explore the potential benefits of betulinol on cognitive function in PSCI, a series of neurobehavioral tests were performed. In the nest building test, the nest building score of PSCI group was significantly lower than that of the normal wild type (WT) group, while the score of betulinol-treated mice was significantly higher than that of PSCI mice (P<0.05, Figure 1 A). In the object recognition test, the discrimination index of PSCI mice was significantly lower than that of the WT group, while betulinol treatment restored it (P<0.01, Figure 1 B); in addition, the latency of the mice to find the target frame gradually shortened on days 1, 2, and 3; on day 4, the latency of betulinol-treated mice was significantly shorter than that of PSCI mice (P<0.05, Figure 1 C); compared with PSCI mice, betulinol-treated mice had significantly increased residence time in the target quadrant (P<0.01, Figure 1 D). These results suggest that betulinol can alleviate cognitive impairment in mice with post-stroke cognitive impairment.
[0029] 2. Betulinol improved the neuropathological changes in mice with post-stroke cognitive impairment.
[0030] To evaluate the effect of betulinol on the pathological changes of post-stroke cognitive impairment, immunostaining of the neuronal marker NeuN was performed; as Figure 2 shown, the immunostaining results of NeuN showed that the neuronal damage in betulinol-treated mice was significantly improved compared with PSCI mice; these results suggest that betulinol treatment can alleviate the neuropathological changes in mice with post-stroke cognitive impairment.
[0031] 3. Betulinol can inhibit neuroinflammation in post-stroke cognitive impairment
[0032] Neuroinflammation is believed to be involved in the pathogenesis of post-stroke cognitive impairment. To explore the role of betulinol in regulating neuroinflammation after post-stroke cognitive impairment, the activation of microglia in mice with post-stroke cognitive impairment was evaluated using immunofluorescence staining analysis; Iba1 is a marker of activated microglia; as Figure 3 shown, the expression level of Iba1 in the betulinol treatment group was lower than that in the PSCI group, suggesting that betulinol treatment can significantly inhibit neuroinflammation in post-stroke cognitive impairment.
[0033] Conclusion:
[0034] The present application has obvious advantages of improving the cognitive impairment and neuron damage of post-stroke cognitive impairment, and the betulin of the present application can be used as a drug for preventing and treating post-stroke cognitive impairment, and also can be used as an auxiliary treatment for preventing and treating post-stroke cognitive impairment in clinic. The present application has good social benefits and is worthy of clinical popularization and application. In summary, the present application develops a drug for preventing and treating post-stroke cognitive impairment with good curative effect and no toxic side effect, and the animal experiment research proves that the betulin preparation can play a role in preventing and treating post-stroke cognitive impairment.
[0035] The preferred embodiments of the present application are merely used for limiting the scope of the present application, and any equivalent changes and modifications made according to the scope of the patent application of the present application are covered by the scope of the present application. Since the present application discloses, for the first time, the application of betulin as a pharmaceutically active ingredient in the preparation of a drug for treating post-stroke cognitive impairment, therefore, as long as the drug is used for treating post-stroke cognitive impairment, the drug prepared by combining with excipients belongs to the protection scope of the present application.
Claims
1. Use of betulin in improving post-stroke cognitive impairment composition, the composition includes medicine, health care product and beverage.
2. The application of betulinol in the composition for improving post-stroke cognitive impairment according to claim 1, characterized in that... Betulin refers to a natural triterpenoid compound extracted from white birch bark.
3. The application of betulinol in the composition for improving post-stroke cognitive impairment according to claim 1, characterized in that... The betulin composition has the effect of inhibiting neuroinflammation of post-stroke cognitive impairment.