Application of dyclonine and / or derivative thereof in preparation of medicine for treating post-resuscitation brain injury
By using dacronin to prepare a drug for treating brain injury after resuscitation, the treatment challenge of ischemic brain injury after cardiac arrest has been solved, achieving the effect of effectively protecting brain function and reducing economic burden.
Patent Information
- Application Number
- CN202510912997.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-03
- Publication Date
- 2025-11-04
AI Technical Summary
There is a lack of effective drug treatments to address brain ischemia-reperfusion injury caused by resuscitation after cardiac arrest, and hypothermia therapy is costly and has complications, making it difficult to apply widely.
Drugs for treating post-recovery brain injury can be prepared using dacronin and/or its derivatives, which can protect brain function by improving neuronal and microglial cell viability and reducing economic burden.
Dacronin significantly improved cell damage caused by hypoglycemia and hypoxia, increased the survival rate of rats undergoing cardiopulmonary resuscitation after cardiac arrest, protected brain nerve function, reduced nerve cell and mitochondrial damage, and increased ATP levels.
Smart Images
Figure CN120884591A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of medicine, in particular to the application of dyclonine and / or its derivatives in the preparation of drugs for improving the treatment of post-resuscitation brain injury. BACKGROUND
[0002] Cardiac arrest (CA) is a serious cardiovascular emergency that threatens human life, with a very high mortality and disability rate, and is an important cause of death worldwide. In the United States, the number of patients with cardiac arrest is 30-37 million per year, and in China, about 1.02 million people suffer from cardiac arrest each year. At present, the only method for rescuing patients with cardiac arrest is cardiopulmonary resuscitation, which refers to the use of artificial or mechanical devices to perform external chest compression and artificial respiration to restore cardiac pumping function, increase cardiac output and coronary perfusion pressure, and reestablish blood circulation to restore spontaneous circulation (ROSC) in patients. In recent years, although the popularity of cardiopulmonary resuscitation and the improvement of the emergency system, more and more patients with cardiac arrest can get early assistance, but the long-term survival rate of patients is still very low, and the 30-day mortality rate is still as high as 90%, the main reason is the damage caused by global cerebral ischemia and hypoxia after resuscitation. After resuscitation, most patients with cardiac arrest have neurological dysfunction, even coma or remain in a vegetative state, causing a huge economic and social burden. In terms of successful resuscitation, not only effective circulation should be restored in time, but also the damage of organs, especially brain tissue, after resuscitation should be overcome. Therefore, the degree of cerebral ischemic injury is the key to determine the prognosis of patients with cardiac arrest.
[0003] The progression of ischemia-reperfusion injury after cardiac arrest to neuronal death is a complex process involving multiple key pathways, including excitotoxicity, mitochondrial dysfunction, oxidative stress, and persistent inflammatory response. Currently, there is no specific intervention measure for the related pathways of the cascade reaction and secondary neuronal injury after cardiac arrest. Although some drugs have shown promising results in preclinical studies, there is currently no clinically approved drug for the treatment of ischemic brain injury.
[0004] Currently, there are very limited treatment means for post-resuscitation cerebral ischemic injury in clinical practice, and hypothermia therapy is the only treatment measure that has been proven to be effective in protecting brain function after resuscitation, and there is a lack of drug treatment means. However, some complications may occur during hypothermia therapy, mainly including: muscle tremor, low immune function, respiratory tract infection, pressure sores, arrhythmia (bradycardia, ventricular premature beat (premature beat), ventricular fibrillation, etc.), circulatory instability (hypotension), rebound intracranial pressure, coagulation dysfunction (low coagulation and bleeding tendency), electrolyte imbalance (high sodium, low potassium, low magnesium, low chloride, low calcium, etc.), and at the same time, vascular hypothermia is very expensive, and ordinary patients are difficult to afford.
[0005] Therefore, it is urgent to develop a drug capable of being applied in treating brain injury after resuscitation. SUMMARY
[0006] The present application aims to overcome the deficiencies of the prior art and provide the use of dyclonine in the preparation of a drug for treating brain injury after resuscitation.
[0007] To achieve the above-mentioned object, the technical scheme adopted by the present application is as follows:
[0008] In a first aspect, the present application provides the use of dyclonine and / or its derivatives in the preparation of a drug for treating brain injury after resuscitation.
[0009] The present application proves by cell experiments that dyclonine can improve the efficacy of reducing the viability of neurons and microglial cells caused by resuming glucose and oxygen after hypoxia and anoxia, so that the viability of brain cells is restored. At the same time, the present application also proves by animal experiments that dyclonine can improve the brain nerve function, the number of Nissl bodies, the structure of mitochondria and the ATP level of rats with global ischemic injury caused by cardiopulmonary resuscitation after cardiac arrest, thereby playing a role in improving brain injury caused by cardiopulmonary resuscitation after cardiac arrest, and dyclonine can also protect brain function. In addition, the cost of taking dyclonine is much lower than that of hypothermia treatment, effectively reducing the economic burden of patients.
[0010] As a preferred embodiment of the use of the present application, the drug for treating brain injury after resuscitation is a drug for treating brain injury after cardiopulmonary resuscitation and / or a drug for treating brain injury after resuscitation from cardiac arrest.
[0011] In a second aspect, the present application provides the use of dyclonine and / or its derivatives in the preparation of a drug for protecting brain function after resuscitation.
[0012] As a preferred embodiment of the use of the present application, the drug for protecting brain function after resuscitation is a drug for protecting brain function after cardiopulmonary resuscitation and / or a drug for protecting brain function after resuscitation from cardiac arrest.
[0013] In a third aspect, the present application provides the use of dyclonine and / or its derivatives in the preparation of a drug for treating ischemic brain injury.
[0014] As a preferred embodiment of the use of the present application, the dyclonine derivative is at least one of a pharmaceutically acceptable salt of dyclonine, an ester derivative of dyclonine, an ether derivative of dyclonine, and a cationic derivative of dyclonine.
[0015] As a preferred embodiment of the use of the present application, the pharmaceutically acceptable salt of dyclonine includes at least one of dyclonine hydrochloride, dyclonine sulfate, dyclonine maleate, dyclonine citrate, and dyclonine camphorsulfonate.
[0016] As a preferred embodiment of the application, the dosage form of the drug for treating post-resuscitation brain injury, protecting post-resuscitation brain function or treating ischemic brain injury is at least one of a capsule, a tablet, a microcapsule preparation, an injection, a suppository, a spray, an ointment, a gel, a solution, a powder, a lotion, a tincture, an oil, a cream and an aerosol.
[0017] In a fourth aspect, the application provides a drug for treating post-resuscitation brain injury, comprising dyclonine and / or a derivative thereof and a pharmaceutically acceptable excipient.
[0018] As a preferred embodiment of the drug of the application, the excipient comprises at least one of a thickening agent, a disintegrating agent and a filler, but is not limited thereto.
[0019] Compared with the prior art, the application has the following beneficial effects:
[0020] The application proves by cell experiments and animal experiments that dyclonine has the effects of improving cell damage caused by glucose deprivation / hypoxia / glucose supply / reoxygenation and whole brain ischemic injury caused by cardiac arrest / cardiac arrest, and can be used for treating brain injury caused by cardiac arrest / cardiac arrest, thereby playing a role in protecting brain nerves and brain function. BRIEF DESCRIPTION OF DRAWINGS
[0021] Figure 1 Effects of different treatments on cell viability of glucose deprivation / hypoxia / glucose supply / reoxygenation HT22 cells (A) and BV2 cells (B) in Experimental Example 1 of the application;
[0022] Figure 2 Effects of different treatment groups on survival rate of rats with whole brain ischemic injury caused by cardiac arrest / cardiac arrest in Experimental Example 2 of the application;
[0023] Figure 3 Effects of different treatments on neurological function of rats with whole brain ischemic injury caused by cardiac arrest / cardiac arrest in Experimental Example 2 of the application;
[0024] Figure 4 Effects of different treatments on brain nerve cell injury of rats with whole brain ischemic injury caused by cardiac arrest / cardiac arrest in Experimental Example 2 of the application, wherein A is the observation result of Nissl staining, and B is the quantitative analysis result of Nissl staining;
[0025] Figure 5 Effects of different treatments on brain mitochondrial structure of rats with whole brain ischemic injury caused by cardiac arrest / cardiac arrest in Experimental Example 2 of the application;
[0026] Figure 6 Effects of different treatments on brain ATP level of rats with whole brain ischemic injury caused by cardiac arrest / cardiac arrest in Experimental Example 2 of the application;
[0027] In the above figures, "*", "**" or "***" indicates a significant difference between the two groups (P < 0.05, P < 0.01 or P < 0.001) ; with "ns" indicates no significant difference between the two groups;
[0028] In the above Figure 1 , Control represents the blank control group, OGD / OGR represents the administration of 0 μM dyclonine, DCN10 represents the administration of 10 μM dyclonine, and DCN50 represents the administration of 50 μM dyclonine;
[0029] In the above Figures 2-6 , DCN2 represents the administration of 2 mg / kg dyclonine group (low dose group), and DCN10 represents the administration of 10 mg / kg dyclonine group (high dose group). DETAILED DESCRIPTION
[0030] In order to better illustrate the purpose, technical scheme and advantages of the present application, the present application will be further described below in combination with specific examples.
[0031] Other materials, reagents, etc. used in the examples can be obtained from commercial channels unless otherwise specified.
[0032] The technologies not described in detail in the following examples, comparative examples and effect examples are all common technologies in the art, which can refer to "Experimental Manual of Molecular Biology" (Ma Wenli, People's Military Medical Publishing House), "Experimental Manual of Molecular Biology (Second Edition) " (Zhejiang University Press), "Experimental Manual of Cell Biology" (Yang Hongbin, Hou Lixia, Zhang Yuxi, Higher Education Press).
[0033] In the following experimental examples, dyclonine was purchased from MedChemExpress Company, and the item number was HY-B0364A;
[0034] Nissl staining solution was purchased from Shanghai Biyun Tian Biotechnology Co., Ltd., and the item number was C0117;
[0035] ATP detection kit was purchased from Shanghai Biyun Tian Biotechnology Co., Ltd., and the item number was S0026.
[0036] Experimental Example 1
[0037] In order to verify whether dyclonine has the effect of treating brain injury after resuscitation in vitro, a cell glucose deprivation / hypoxia / glucose reperfusion model was constructed for evaluation, and the specific scheme was as follows:
[0038] Firstly, the sugar-free Earle's balanced solution was saturated with 95% N2+5% CO2 for 2h to remove oxygen. Then the cultured cells were replaced with sugar-free Earle's balanced solution after hypoxic treatment, and the cells were placed in a hypoxic incubator (37℃, 5% CO2, 95% N2) for culture. After 8h of sugar and oxygen deprivation culture of neuron cells HT22 or microglial cells BV2, the cells were removed, the normal culture medium containing 0μM, 10μM or 50μM dyclonine was replaced, and the cells were placed in a normal incubator (37℃, 20% O2, 5% CO2) for culture. After 12h of reoxygenation, the cell activity was observed and detected. HT22 cells and BV2 cells were seeded in 96-well plates at a density of 1×10 4 6 / well as blank control group. After the treatment of each group of cells, the original culture medium was discarded, and the cells were washed with PBS for 3 times. 100μL of serum-free DMEM and 10μL of CCK-8 solution were added to each well, and the cells were incubated at 37℃ for 2h. The absorbance value (OD value) was measured at 450nm by a microplate reader. The cell activity was calculated by the ratio of the OD value of each group to the OD value of the normal control group. The results are shown in Figure 1 .
[0039] As shown in Figure 1 , the activity of HT22 cells ( Figure 1 A) and BV2 cells ( Figure 1 B) was significantly reduced after sugar and oxygen deprivation / reoxygenation. However, after treatment with relatively high concentration of dyclonine (50μM), the activity of HT22 cells ( Figure 1 A) and BV2 cells ( Figure 1 B) was significantly increased, indicating that dyclonine can improve the phenomenon of reduced activity of nerve cells after sugar and oxygen deprivation / reoxygenation treatment.
[0040] Experimental Example 2
[0041] In order to verify whether dyclonine also has the effect of treating brain injury after resuscitation in vivo, a cell sugar and oxygen deprivation / reoxygenation model was constructed for evaluation, and the specific scheme is as follows:
[0042] 1. Constructing an animal model of global cerebral ischemic injury caused by cardiac arrest / cardio-pulmonary resuscitation.
[0043] SPF level healthy male SD rats (330-350g) were fasted for 12h before model establishment, and then anesthetized with 45mg / kg sodium pentobarbital, and a 14-gauge tracheal tube was inserted directly into the oral cavity. A 23-gauge PE-50 polyethylene tube was inserted into the left femoral artery to measure aortic pressure. After recording the baseline hemodynamic data, the trachea was occluded for about 11 min, including 6 min of complete cardiac arrest (mean arterial pressure ≤20 mmHg). After 11 min of asphyxia, an animal cardiopulmonary resuscitation device developed by the laboratory was used to start chest compression at a frequency of 250 times / min and synchronous mechanical ventilation (oxygen concentration 100%) at a frequency of 100 times / min. The compression depth was 1 / 3 of the anteroposterior diameter of the thorax, and the tidal volume was 0.6 mL / 100g body weight. After 2 min of compression, 0.04 mg / kg of adrenaline was administered. After 4 min of chest compression, if the heart was in fibrillation, up to 3 times of 2J biphasic wave defibrillation was given. If spontaneous circulation was not restored, chest compression and mechanical ventilation were restarted for 30s until spontaneous circulation was restored. If spontaneous circulation was not restored after 3 cycles, the resuscitation was abandoned. Spontaneous circulation was defined as the restoration of supraventricular rhythm, mean arterial pressure ≥60 mmHg and maintained for more than 5 min. After successful restoration of spontaneous circulation, mechanical ventilation was given with 100% oxygen for 15 min, 70% oxygen for 15 min, 50% oxygen for 15 min, and 30% oxygen for 15 min. Mechanical ventilation was stopped after 1h, and hemodynamics was continuously monitored for 2h. After 2h, all catheters were removed, and the rats were returned to the cage for feeding.
[0044] After the rats were resuscitated, the rats were randomly divided into 4 groups: sham operation group, resuscitation control group, low-dose drug group, and high-dose drug group. The rats in the sham operation group only underwent surgery and were not induced to cardiac arrest and resuscitation. The rats in the resuscitation control group underwent surgery and were induced to cardiac arrest and resuscitation, but were not given any drug. The rats in the drug treatment groups were given drugs through intraperitoneal injection 5 minutes after resuscitation. The rats in the low-dose group were given 2mg / kg dyclonine, and the rats in the high-dose group were given 10mg / kg dyclonine.
[0045] The rats in the above groups were evaluated for the following indicators:
[0046] (1) Survival rate of rats after resuscitation. The survival time and survival rate of rats after resuscitation were counted, and the results are shown in Figure 2 .
[0047] As shown in Figure 2 , the rats were treated with dyclonine at two doses (high dose, 10mg / kg; low dose, 2mg / kg) after resuscitation, and the survival rate of rats was counted for 3 days. The results showed that compared with the resuscitation control group, high-dose dyclonine treatment significantly improved the survival rate of rats after resuscitation, indicating that dyclonine has the effect of reducing the mortality rate after cardiopulmonary resuscitation.
[0048] Neurological deficit score. The neurological deficit score of rats was detected according to the method described by Geocadin et al. (Geocadin RG, Ghodadra R, Kimura T, Lei H, Sherman DL, Hanley DF, Thakor NV. A novel quantitative EEG injury measure of global cerebral ischemia. Clin Neurophysiol. 2000; 111(10): 1779-87). The neurological deficit score was 0-80 points, and the score included overall behavior, brainstem function, motor evaluation, sensory evaluation, motor behavior, behavior, and seizure evaluation. 0 points corresponded to brain death, and 80 points corresponded to no brain injury. The results are shown in Figure 3 .
[0049] As shown in Figure 3 , compared with the sham operation group, the score of the resuscitation control group was significantly reduced, indicating that the brain injury model was successfully constructed after cardiopulmonary resuscitation after cardiac arrest. Compared with the resuscitation control group, the neurological score of rats treated with high-dose dyclonine was significantly increased, indicating that dyclonine had a protective effect on the neurological function of the brain after cardiopulmonary resuscitation.
[0050] (2) Nissl staining. After the rats were sacrificed, the brain tissue was taken out to prepare paraffin sections, and the Nissl staining solution was operated according to the instructions. The cell staining was observed under an optical microscope, and the results are shown in Figure 4 .
[0051] As shown in Figure 4 , compared with the sham operation group, the number of Nissl bodies in the nerve cells of the resuscitation control group rats was significantly reduced, and the number of Nissl bodies in the nerve cells of rats treated with high-dose dyclonine was significantly increased compared with the resuscitation control group, which indicated that high-dose dyclonine treatment significantly reduced the nerve cell damage of rats after resuscitation.
[0052] (3) Electron microscopy. The rat brain hippocampal tissue was fixed with 2.5% v / v glutaraldehyde, then dehydrated, permeated, embedded, sectioned and stained, and the mitochondrial damage of neurons was observed by transmission electron microscopy. The results are shown in Figure 5 .
[0053] As shown in Figure 5 , the mitochondrial electron density in the brain of rats after resuscitation was reduced, and the mitochondria were vacuolated, indicating that the mitochondria were severely damaged, and high-dose dyclonine treatment significantly reduced the mitochondrial damage in the nerve cells of rats after resuscitation, indicating that dyclonine could improve the mitochondrial damage in the nerve cells of rats after resuscitation.
[0054] (4) ATP detection. The content of ATP in the brain was measured by using an ATP measurement kit, and the results are shown inFigure 6 .
[0055] As shown in Figure 6 , compared with the sham operation group, the brain ATP content of the resuscitation control group was significantly reduced, and compared with the resuscitation control group, the brain ATP content of the rats treated with high-dose dyclonine was significantly increased, indicating that dyclonine has the effect of improving the decrease of brain ATP content caused by cardiac arrest.
[0056] In summary, dyclonine has the efficacy of treating brain injury after cardiac arrest / cardiac arrest, can also protect brain function after cardiopulmonary resuscitation, and can also be applied in the treatment of ischemic brain injury.
[0057] Finally, it should be explained that the above examples are only used to illustrate the technical solutions of the present application and not to limit the protection scope of the present application. Although the present application has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present application can be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present application.
Claims
1. Use of dyclonine and / or a derivative thereof in the preparation of a medicament for treating brain injury after resuscitation.
2. The use according to claim 1, characterized in that, The medicament for treating brain injury after resuscitation is a medicament for treating brain injury after cardiopulmonary resuscitation and / or a medicament for treating brain injury after resuscitation from cardiac arrest.
3. Use of dyclonine and / or a derivative thereof in the preparation of a medicament for protecting brain function after resuscitation.
4. The use according to claim 3, characterized in that, The medicament for protecting brain function after resuscitation is a medicament for protecting brain function after cardiopulmonary resuscitation and / or a medicament for protecting brain function after resuscitation from cardiac arrest.
5. Use of dyclonine and / or a derivative thereof in the preparation of a medicament for treating ischemic brain injury.
6. Use according to any one of claims 1 to 5, characterized in that, The dyclonine derivative is at least one of a pharmaceutically acceptable salt of dyclonine, an ester derivative of dyclonine, an ether derivative of dyclonine, and a cationic derivative of dyclonine.
7. The use according to claim 6, characterized in that, The pharmaceutically acceptable salt of dyclonine includes at least one of dyclonine hydrochloride, dyclonine sulfate, dyclonine maleate, dyclonine citrate, and dyclonine camsylate.
8. Use according to any one of claims 1 to 5, wherein The dosage form of the medicament for treating brain injury after resuscitation, the medicament for protecting brain function after resuscitation, or the medicament for treating ischemic brain injury is at least one of a capsule, a tablet, a microcapsule preparation, an injection, a suppository, a spray, an ointment, a gel, a solution, a powder, a lotion, a tincture, an oil, a cream, and an aerosol.
9. A medicament for treating post-resuscitation brain injury, characterized by, The dyclonine and / or a derivative thereof and a pharmaceutically acceptable excipient are included.