Medicine for preventing cerebral ischemia reperfusion injury and preparation method thereof

Abstract

The invention discloses a medicine for preventing cerebral ischemia reperfusion injury and a preparation method thereof, and relates to the technical field of cerebral ischemia reperfusion injury prevention. Comprising the following steps: S1, screening fresh raw material ginkgo leaves to remove damaged incomplete defective products with insect pest traces, and crushing the ginkgo leaves into powder by using a crusher; sa1, after the kudzu vine roots are cut into blocks and boiled, drying and smashing are conducted, then 95% ethyl alcohol is mixed into the kudzu vine root powder according to the proportion of 1: 2, and then heating is conducted for two hours at the temperature of 75 DEG C; the ginkgo biloba extract contains ginkgo flavone, bilobalide and other active ingredients capable of preventing cerebral ischemia reperfusion injury, bilobalide can reduce neuronal apoptosis and necrosis in the cerebral ischemia reperfusion process, puerarin can also play a role in scavenging free radicals and inhibiting oxidation reaction, and can also play a role in preventing cerebral ischemia reperfusion injury. And the two are matched for use, so that a plurality of links of cerebral ischemia reperfusion which may cause injury to the brain can be covered, and brain tissues are more effectively prevented from being injured.

Description

Technical Field

[0001] The invention relates to the technical field of preventing cerebral ischemia-reperfusion injury, and in particular to a medicine for preventing cerebral ischemia-reperfusion injury and a preparation method thereof. Background Art

[0002] The brain is the most sensitive organ in the human body to hypoxia, and tissue damage caused by cerebral ischemia is the main cause of fatal diseases. Cerebral ischemia reperfusion injury refers to the damage to the structure and function of brain tissue caused by ischemia. During the rescue and treatment of cerebral ischemic diseases, the main factor causing tissue damage is not ischemia itself, but the damage caused by reactive oxygen free radicals to microvessels and brain parenchyma organs when blood re-enters the ischemic brain tissue after blood supply is restored. The synthesis capacity of antioxidant enzymes that can scavenge free radicals in ischemic tissue is impaired, thereby aggravating the damage of free radicals to tissues after ischemia and reperfusion. In addition, when blood re-enters the brain, a large amount of calcium ions will flow in, causing damage to tissue cells; neutrophils can also cause damage to the patient's organs and tissues through oxygen free radicals, causing inflammatory reactions.

[0003] The clinical manifestations of cerebral ischemia-reperfusion injury are generally dizziness, headache, nausea, drowsiness, memory loss, language disorders, etc. In severe cases, local blood vessels may rupture and compress the soft tissue of the brain. Therefore, drugs are needed to restore blood flow as soon as possible during cerebral ischemia, reduce ischemia time, control blood reperfusion conditions, improve ischemic tissue metabolism and remove free radicals to prevent damage during reperfusion.

[0004] After searching, the Chinese patent publication number CN116942752B discloses a Dai medicine composition for preventing cerebral ischemia-reperfusion injury, its preparation method and application, and the raw materials include: 30-100 parts of dragon's blood, 0-50 parts of calamus and 5-20 parts of natural borneol. Although the Dai medicine used in the above patent has the effects of clearing heat and opening the orifices, removing wind and unblocking the meridians, and removing blood stasis and relieving pain, its specific active ingredients are unknown, the active ingredients have not been purified, the efficacy is insufficient, and the preventive and protective effect on brain neuron damage is insufficient. Summary of the invention

[0005] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a medicine for preventing cerebral ischemia-reperfusion injury and a preparation method thereof.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for preparing a drug for preventing cerebral ischemia-reperfusion injury comprises the following steps:

[0008] S1: Screen the fresh raw ginkgo leaves to remove those with traces of pests and damaged, incomplete and defective products, and use a grinder to grind the ginkgo leaves into powder;

[0009] Sa1: After the kudzu root is cut into pieces and boiled, it is dried and crushed, and then 95% ethanol is mixed into the kudzu root powder in a ratio of 1:2, and then heated at 75°C for two hours; after the heating is completed, the mixture is evaporated under reduced pressure to recover the ethanol, and the kudzu root powder is obtained;

[0010] S2: First, the ginkgo leaf powder crushed in step S1, the kudzu root powder obtained in step Sa1 and the entrainer are mixed and then put into the extraction kettle of the supercritical extraction device, wherein the entrainer amount mixed into the ginkgo leaf powder is 5% by weight of the ginkgo leaf powder and the kudzu root powder;

[0011] An ultrasonic generator can be installed on the outside of the extraction kettle;

[0012] Then, the gas in the carbon dioxide cylinder is introduced into the extraction kettle of the supercritical extraction device, the carbon dioxide gas is preheated to about 31.2°C, and then the high-pressure pump is started to pressurize the carbon dioxide gas;

[0013] The mixture was then subjected to three-stage extraction in an extraction kettle;

[0014] S3: After the extraction is completed, the mixture in the extraction kettle is moved to the separation kettle, and the carbon dioxide in the supercritical fluid state gradually changes into a gaseous state and is separated from the extract;

[0015] S4: The desolvated solution is then filtered and evaporated to dryness under reduced pressure. The pressure of the reduced pressure evaporation is preferably 1.2 KPa and the temperature is 45° C., until a powder containing a mixture of ginkgo flavonoids, ginkgo lactones and puerarin is finally obtained.

[0016] Preferably, the powder particle size of the ginkgo leaf and kudzu root in steps S1 and Sa1 is 0.18 mm-0.22 mm.

[0017] Furthermore: the entrainer used in step S2 is a mixture of 95% methanol and 95% ethanol in a ratio of 8:2, wherein the 95% ethanol can be the ethanol recovered in step Sa1.

[0018] Further: the three-stage extraction in step S2 is:

[0019] In the first stage, the extraction temperature was raised to 40°C and the extraction was carried out for 15 minutes;

[0020] Then, the temperature in the extraction kettle is further increased to 43° C., and an entrainer having a weight of 5% of the weight of the ginkgo leaf powder and the kudzu root powder is continuously sprayed on the ginkgo leaf powder, and the ultrasonic generator outside the extraction kettle is started to perform ultrasonic vibration on the internal mixture to further increase the extraction rate, and the second extraction is performed after 45 minutes of extraction;

[0021] Then, the ultrasonic generator is turned off. Since the temperature in the extraction kettle will rise during ultrasonic vibration extraction, the temperature needs to be increased after stopping the ultrasonic vibration, preferably to 46° C. and maintained for 60 minutes for the third stage of extraction.

[0022] As a preferred solution of the present invention: the ultrasonic generator preferably has a power of 500W and a frequency of 50kHz; the ultrasonic generator vibrates through 4 probes evenly distributed below the extraction kettle.

[0023] As a further solution of the present invention: the supercritical fluidization extraction pressure in the S2 step is 25 MPa.

[0024] As a further solution of the present invention: in the step S3, the internal pressure of the separation kettle is reduced to 7 MPa during operation, and the temperature inside the separation kettle is increased to 55°C.

[0025] On the basis of the above scheme: in the step S4, a 0.45 μm microporous filter membrane is used for filtration.

[0026] Based on the above scheme: the weight ratio of the ginkgo leaf raw material to the kudzu root raw material is 2:1.

[0027] A medicine for preventing cerebral ischemia-reperfusion injury, wherein the effective ingredients include ginkgo leaf extract and puerarin.

[0028] The beneficial effects of the present invention are:

[0029] 1. A drug for preventing cerebral ischemia-reperfusion injury. Ginkgo extract contains active ingredients such as ginkgo flavonoids and ginkgolides that can prevent cerebral ischemia-reperfusion injury. Ginkgo lactones can reduce neuronal apoptosis and necrosis during cerebral ischemia-reperfusion. Puerarin can also scavenge free radicals and inhibit oxidative reactions. The combined use of the two can cover multiple links that may cause damage to the brain due to cerebral ischemia-reperfusion, and more effectively avoid damage to brain tissue.

[0030] 2. A method for producing a drug for preventing cerebral ischemia-reperfusion injury, which can effectively reduce processing costs by subjecting crushed ginkgo leaf powder and pretreated kudzu root powder to supercritical fluid extraction.

[0031] 3. A method for manufacturing a drug for preventing cerebral ischemia-reperfusion injury, which further improves the extraction effect of the effective ingredients of Ginkgo and Pueraria lobata by using an ultrasonic generator to assist in ultrasonic extraction during the second stage of extraction, reduces raw material waste, and increases the output of the final product. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The present invention is a schematic flow chart of a method for producing a drug for preventing cerebral ischemia-reperfusion injury. DETAILED DESCRIPTION

[0033] The technical solution of this patent is further described in detail below in conjunction with specific implementation methods.

[0034] The embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0035] Embodiment 1:

[0036] A method for producing a drug for preventing cerebral ischemia-reperfusion injury, such as Figure 1 As shown, the following steps are included:

[0037] S1: Screening fresh raw ginkgo leaves to remove those with traces of pests and damaged, incomplete and defective products, then washing and drying, and then using a grinder to grind the ginkgo leaves into powder. The preferred ginkgo leaf powder has a particle size of 0.18 mm-0.22 mm. After grinding, use an 80-mesh screen to screen. If the ginkgo leaf powder is too coarse, it will affect the extraction rate. If the powder is too fine, the mass transfer resistance will increase, which is not conducive to extraction.

[0038] Sa1: After boiling, kudzu root pieces with a weight of 50% of that of ginkgo leaf pieces are dried and crushed. The particle size of the kudzu root powder is the same as that of the ginkgo leaf powder. The dried kudzu root powder is then sieved through an 80-mesh sieve, and then 95% ethanol is mixed into the kudzu root powder at a ratio of 1:2, and then heated at 75°C for two hours. After the heating is completed, the mixture is evaporated under reduced pressure to recover the ethanol, thereby obtaining kudzu root powder;

[0039] S2: First, the ginkgo leaf powder crushed in step S1, the kudzu root powder obtained in step Sa1 and the entrainer are mixed and then put into the extraction kettle of the supercritical extraction device. The entrainer is preferably a mixture of 95% methanol and 95% ethanol in a ratio of 8:2, wherein the 95% ethanol can be the ethanol recovered in step Sa1. The amount of entrainer mixed into the ginkgo leaf powder here is 5% of the weight of the ginkgo leaf powder and the kudzu root powder;

[0040] Preferably, an ultrasonic generator can be installed on the outside of the extraction kettle to improve the extraction effect. The ultrasonic generator vibrates through 4 probes evenly distributed under the extraction kettle.

[0041] Then the gas in the carbon dioxide cylinder is introduced into the extraction kettle of the supercritical extraction device, the carbon dioxide gas is preheated to about 31.2°C, and then the high-pressure pump is started to pressurize the carbon dioxide gas to 25MPa. Higher pressure can improve the extraction rate, and at the same time has better extraction selectivity and higher removal rate;

[0042] Then the temperature in the extraction kettle was further raised to 40°C, and the extraction was carried out for 15 minutes to carry out the first stage of extraction;

[0043] Then, the temperature in the extraction kettle is further increased to 43° C., and an entrainer having a weight of 5% of the weight of the ginkgo leaf powder and the kudzu root powder is continuously sprayed on the ginkgo leaf powder, and the ultrasonic generator outside the extraction kettle is started to perform ultrasonic vibration on the internal mixture to further improve the extraction rate. The ultrasonic generator preferably has a power of 500 W and a frequency of 50 kHz. The second extraction is performed after 45 minutes of extraction.

[0044] Then, the ultrasonic generator is turned off. Since the temperature in the extraction kettle will increase during ultrasonic vibration extraction, the temperature needs to be increased after the ultrasonic vibration is stopped. Preferably, the temperature is increased to 46° C. and maintained for 60 minutes for the third extraction. In the three-stage extraction process, the second extraction has the highest amount of ginkgolide, and the extraction amounts of the first, second and third extractions are 26.8%, 64.8% and 8.4% of the total extraction amount, respectively.

[0045] S3: After the extraction is completed, the mixture in the extraction kettle is moved to the separation kettle, and then the pressure in the separation kettle is reduced to 7MPa, and the temperature in the separation kettle is increased to 55°C. At this time, the carbon dioxide in the supercritical fluid state will gradually change into a gaseous state and be separated from the extract. The carbon dioxide gas and methane and ethane gases at the separation site can be recovered after separation and condensation;

[0046] S4: The desolvated solution is then filtered, preferably using a 0.45 μm microporous filter membrane, and then dried by reduced pressure evaporation. The pressure of the reduced pressure evaporation is preferably 1.2 KPa and the temperature is 45° C., until a powder containing a mixture of ginkgo flavonoids, ginkgo lactones and puerarin is finally obtained.

[0047] Ginkgo extract contains active ingredients such as ginkgo flavonoids and ginkgolides that can prevent cerebral ischemia-reperfusion injury. These ingredients have good antioxidant and anti-inflammatory effects, and can effectively increase the activity of antioxidant enzymes SOD and GSH-Px content in the brain of ischemic animals, reduce lipid peroxidation, and reduce the damage to brain tissue after cerebral ischemia-reperfusion.

[0048] Ginkgolide is a natural platelet-activating factor receptor antagonist that can effectively inhibit abnormal platelet aggregation and thrombosis, improve the tolerance of brain tissue to ischemia, and improve blood circulation to allow the brain tissue to obtain more adequate blood and oxygen supply. Ginkgolide can also reduce neuronal apoptosis and necrosis during cerebral ischemia-reperfusion, and has a good effect in protecting brain nerves.

[0049] At the same time, Ginkgo extract can inhibit matrix metalloproteinases (MMPs) to reduce their destructive effects on the blood-brain barrier and protect the integrity of the blood-brain barrier. It can also regulate aquaporin 4 (AQP4) to maintain the water balance of the blood-brain barrier and prevent pathological phenomena such as edema after cerebral ischemia-reperfusion.

[0050] Puerarin can also scavenge free radicals and inhibit oxidative reactions, while it can reduce the levels of plasma malondialdehyde and endothelin during cerebral ischemia-reperfusion, promote the release of calcitonin gene-related peptide, and increase the activity of superoxide dismutase, thereby reducing the damage of ischemia-reperfusion to nerve cells and further improving the protective effect on brain nerve cells. The combined use of the two can cover multiple links that may cause damage to the brain due to cerebral ischemia-reperfusion, and more effectively avoid damage to brain tissue.

[0051] During the production process, the processing cost can be effectively reduced by subjecting the crushed ginkgo leaf powder and the pre-treated kudzu root powder to supercritical fluid extraction. In addition, by using an ultrasonic generator to assist in ultrasonic extraction in the second stage of extraction, the extraction effect of the effective ingredients of ginkgo and kudzu root is further improved, the waste of raw materials is reduced, and the output of the final product is increased. The final powder can be made into tablets or capsules for reuse to prevent cerebral ischemia-reperfusion damage.

[0052] The above is a preferred specific implementation manner of the present invention, and the protection scope of the present invention is not limited thereto. Any modification, equivalent substitution and improvement made by any technician familiar with the field within the technical scope disclosed by the present invention in combination with the prior art or public common sense, within the spirit and principle of the present invention, shall be covered by the protection scope of the present invention.

Claims

1. A method for preparing a drug for preventing cerebral ischemia-reperfusion injury, comprising the following steps: S1: Screen the fresh raw ginkgo leaves to remove those with traces of pests and damaged, incomplete and defective products, and use a grinder to grind the ginkgo leaves into powder; Sa1: After the kudzu root is cut into pieces and boiled, it is dried and crushed, and then 95% ethanol is mixed into the kudzu root powder in a ratio of 1:2, and then heated at 75°C for two hours; after the heating is completed, the mixture is evaporated under reduced pressure to recover the ethanol, and the kudzu root powder is obtained; S2: First, the ginkgo leaf powder crushed in step S1, the kudzu root powder obtained in step Sa1 and the entrainer are mixed and then put into the extraction kettle of the supercritical extraction device, wherein the entrainer amount mixed into the ginkgo leaf powder is 5% by weight of the ginkgo leaf powder and the kudzu root powder; An ultrasonic generator can be installed on the outside of the extraction kettle; Then, the gas in the carbon dioxide cylinder is introduced into the extraction kettle of the supercritical extraction device, the carbon dioxide gas is preheated to about 31.2°C, and then the high-pressure pump is started to pressurize the carbon dioxide gas; The mixture was then subjected to three-stage extraction in an extraction kettle; S3: After the extraction is completed, the mixture in the extraction kettle is moved to the separation kettle, and the carbon dioxide in the supercritical fluid state gradually changes into a gaseous state and is separated from the extract; S4: The desolvated solution is then filtered and evaporated to dryness under reduced pressure. The pressure of the reduced pressure evaporation is preferably 1.2 KPa and the temperature is 45° C., until a powder containing a mixture of ginkgo flavonoids, ginkgo lactones and puerarin is finally obtained.

2. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 1, characterized in that: In the steps S1 and Sa1, the powder particle size of the ginkgo leaf and kudzu root is 0.18 mm-0.22 mm.

3. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 2, characterized in that: The entrainer used in step S2 is a mixture of 95% methanol and 95% ethanol in a ratio of 8:2, wherein the 95% ethanol may be the ethanol recovered in step Sa1.

4. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 3, characterized in that: The three-stage extraction in step S2 is: In the first stage, the extraction temperature was raised to 40°C and the extraction was carried out for 15 minutes; Then, the temperature in the extraction kettle is further increased to 43° C., and an entrainer having a weight of 5% of the weight of the ginkgo leaf powder and the kudzu root powder is continuously sprayed on the ginkgo leaf powder, and the ultrasonic generator outside the extraction kettle is started to perform ultrasonic vibration on the internal mixture to further increase the extraction rate, and the second extraction is performed after 45 minutes of extraction; Then, the ultrasonic generator is turned off. Since the temperature in the extraction kettle will rise during ultrasonic vibration extraction, the temperature needs to be increased after stopping the ultrasonic vibration, preferably to 46° C. and maintained for 60 minutes for the third stage of extraction.

5. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 4, characterized in that: The ultrasonic generator preferably has a power of 500W and a frequency of 50kHz; the ultrasonic generator vibrates through 4 probes evenly distributed below the extraction kettle.

6. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 5, characterized in that: The supercritical fluid extraction pressure in step S2 is 25 MPa.

7. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 1, characterized in that: In the step S3, the internal pressure of the separation kettle is reduced to 7 MPa during operation, and the temperature inside the separation kettle is increased to 55°C.

8. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 7, characterized in that: In the step S4, a 0.45 μm microporous filter membrane is used for filtration.

9. The method for producing a drug for preventing cerebral ischemia-reperfusion injury according to claim 1, characterized in that: The weight ratio of the ginkgo leaf raw material to the kudzu root raw material is 2:

1.

10. A drug for preventing cerebral ischemia-reperfusion injury according to any one of claims 1 to 8, wherein the active ingredients include ginkgo leaf extract and puerarin.

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