WO2018113027A1 - Application of bilobalide as synergist in preparation of drugs for preventing cranial nerve injury diseases - Google Patents

Abstract

An application of bilobalide as a synergist in preparation of drugs or health-care products for preventing cranial nerve injury diseases. Bilobalide and hydroxyl derivatives thereof can promote various drugs with a therapeutic or health-care function on cranial nerve injury diseases to enter brain tissues, including ginsenoside, stibene glucoside, resveratrol, levodopa, edaravone, vinpocetine, nicergoline, citicoline, oxiracetam and the like. The efficacy of the drugs can be improved, and side effects and adverse reactions of the drugs can be reduced.

Description

Application of ginkgolide as a synergist in the preparation of drugs for preventing and treating cranial nerve injury diseases Technical field

The present invention relates to a novel application of a compound and a novel pharmaceutical or nutraceutical composition based on the application, and in particular to the use of ginkgolide as a synergist for the preparation of a medicament or a health care product for preventing and treating cranial nerve injury diseases.

Background technique

Brain diseases related to the central nervous system such as ischemic heart disease, neurodegenerative diseases, such as ischemic stroke, chronic cerebral ischemia, stroke sequelae, senile dementia, Parkinson's disease, Alzheimer's disease Etc. has become an increasingly common disease, and the development of new drugs for the treatment of these diseases has been subject to many factors, the most important of which is how the drug passes through the blood-brain barrier. Basically, 100% of macromolecular drugs, including peptides, recombinant proteins, monoclonal antibodies, RNA interference-based drugs, gene therapy-related drugs, etc., cannot cross the blood-brain barrier, and more than 98% of small molecule drugs cannot pass through. Blood brain barrier. And with the aging of the population, the above-mentioned brain diseases related to the central nervous system will be more and more intensified, so research and development of drugs that promote the permeability of the blood-brain barrier has become a top priority.

The blood-brain barrier is a regulatory interface between capillaries and neural tissue in the brain and spinal cord. It is mainly composed of vascular endothelial cells of the central nervous system, perivascular glial cell processes and basement membrane, and is an important structure for maintaining the stability of the brain environment. Under physiological conditions, the blood-brain barrier can selectively pump harmful or excess substances in the brain out of the brain to keep the internal environment of the brain stable. However, in the treatment of brain diseases, the blood-brain barrier is a major obstacle to prevent the penetration of drugs into the brain, so that the drug can not achieve effective drug treatment concentration and affect the efficacy. The blood-brain barrier includes: 1 blood cerebrospinal fluid barrier (BLB); 2 cerebrospinal fluid brain barrier (LBB); 3 blood-brain barrier (BBB); 4 cerebrospinal fluid-brain Tumor barrier and cerebrospinal fluid-lymphatic barrier. Since the surface area of BBB is about 5000 times the surface area of BLB, BBB is the main barrier to control the entry and exit of exogenous substances into the brain parenchyma.

Traditional Chinese medicine has a long history in the prevention and treatment of cardiovascular and cerebrovascular diseases, and has the advantages of remarkable curative effect, safety, low toxicity and small side effects. For example, ginsenoside extract and extract of Polygonum sinensis are important natural drugs for the treatment of brain diseases. However, their active ingredients are ginsenoside Rb1, Rd, Re, Rg1, Shouwustilbene glycoside, Resveratrol, etc. It is not easy to enter the brain tissue through the blood-brain barrier. In addition, commonly used chemical drugs for the prevention and treatment of cerebral ischemic diseases and neurodegenerative diseases, such as levodopa, edaravone, vinpocetine, nigralin, citicoline, oxiracetam, etc. A small amount of drugs can pass through the blood-brain barrier, and in the brain, an effective drug treatment concentration is often not obtained, which affects the therapeutic effect. Therefore, solving the problem of drugs passing through the BBB has become a key and research hotspot in the treatment of brain diseases.

Adenosine Receptor (AR) is a protein molecule that has been found to be extremely important in regulating the permeability of the blood-brain barrier in the tight junction of the blood-brain barrier in recent years. It is an endogenous adenosine molecule that regulates macromolecules through BBB. A new, effective, endogenous blood-brain barrier regulation pathway in the brain is a pathway for promising drugs to enter the CNS in future clinical applications.

After adenosine receptor activation, the expression of tight junction proteins ZO-1, Occludin3 and Claudin-3/5 was significantly down-regulated; in addition, the activity of adenylate cyclase (AC), ion channel and phospholipase was regulated, and phospholipase was activated. C (PLC), PLC activates phosphatidylinositol system to produce inositol triphosphate (IP3) and diglyceride (DG), DAG, DG activates corresponding protein kinase C (PKC) and activates MLC kinase (MLCK), IP3 can promote The release of Ca ²⁺ from the endoplasmic reticulum of the cell interacts with DAG to activate and translocate PKC to tight junctions, directly or indirectly affecting, leading to myosin light chain (MLC), tightly linked structural proteins such as Occludin and ZO- 1. Serine/threonine phosphorylation of cytoskeletal proteins such as ZO-2 and ZO-3, calmodulin binding proteins. Phosphorylation of these proteins mediates the redistribution of cytoskeletal proteins, leading to endothelial cell contraction, changes in cell-cell and cell-basement membrane adhesion states, cell gap formation, increased proliferation, and increased BBB permeability. This results in a decrease in transmembrane resistance, further enhancing BBB permeability, thereby promoting the entry of macromolecules and cells into brain tissue. But did not significantly change its endocytosis. Blocking A1R and A2aR will reduce blood-brain barrier permeability.

Ginkgolide is the only sesquiterpene extracted from the ginkgo biloba ginkgo, and its structural formula is as follows:

The molecular formula is C ₁₅ H ₁₈ O ₈ and its molecular weight is 326.3. It has good curative effect on cardiovascular and cerebrovascular diseases such as angina pectoris, myocardial infarction, coronary heart disease, hypertension and arrhythmia, which can reduce blood pressure, slow heart rate and increase Coronary flow has a protective effect on myocardial ischemia-reperfusion injury, and can also antagonize abnormal platelet aggregation and thrombosis caused by platelet activating factor, thereby reducing plasma viscosity and whole blood viscosity, and protecting nerves. Chinese Patent Application Publication No. CN102670586A discloses the use of ginkgolide in the preparation of a medicament for treating ischemic cardiomyopathy, which regulates cell survival rate, regulates platelet activating factor receptor and platelet activating factor acetylation. Hydrolase and reduction of cardiomyocyte apoptosis in the treatment of ischemic cardiomyopathy. Further, as disclosed in Chinese Patent Publication No. CN101829109B, a new use of albino lactone and its derivatives for the treatment of brain edema, which achieves cerebral edema by regulating aquaporins, is disclosed. Therapeutic effect.

There are no data in the existing literature showing that hydroxy derivatives of bilobalide, biloba lactone extract, bilobalide or biloba lactone extract, especially its glycosides, esters, ether derivatives, etc., have improved blood-brain barrier permeability. Sexuality, a functional report that promotes the entry of drug molecules into brain tissue.

Summary of the invention

The object of the present invention is to provide a new use of ginkgolide, in particular, the use of ginkgolide as a synergist in the preparation of a medicament for preventing and treating cranial nerve damage diseases, wherein the ginkgolide can promote nerve damage diseases of the brain. The therapeutic or health-care drugs pass through the blood-brain barrier and give full play to the drug's efficacy.

Another object of the present invention is to provide a composition for a novel or health care product which can effectively increase the effective concentration of a pharmaceutically active molecule in the brain.

The ginkgo lactone according to the present invention is at least one of a natural product of ginkgolide, a lactone lactone extract, a natural product ginkgolide or a hydroxy derivative of a ginkgolide extract. The ginkgolide extract is at least one of a ginkgolide extract or a ginkgo leaf extract, and the ginkgolide extract has a ginkgolide content of from 1 to 99%. The hydroxy derivative of the natural product ginkgolide or biloba lactone extract is its glycoside, ester or ether.

The inventors have conducted a large number of research experiments, showing that ginkgolide, biloba lactone extract, and its hydroxy derivatives, especially its glycosides, esters, ether derivatives, etc., can effectively improve blood-brain barrier permeability and promote drugs. The molecules enter the brain tissue.

The ginkgolide, the biloba lactone extract, and the hydroxy derivative thereof, particularly the glycoside, ester, and ether derivative thereof, can promote the expression of adenosine receptors in the microvascular endothelium of the brain tissue and inhibit the tight junction protein (ZO). -1 protein and occlaudin protein) expression, improve blood-brain barrier permeability, and promote drug molecules into brain tissue.

The active ingredient in the medicine or health care product for preventing and treating brain nerve damage diseases according to the present invention is selected from the group consisting of natural medicinal active ingredients: ginseng total saponin (containing ginsenoside Rb1, ginsenoside Rd, ginsenoside Re and ginsenoside Rg1, etc.), Shouwu stilbene glycoside, resveratrol, etc., or chemical components: levodopa, edaravone, vinpocetine, nigralin, citicoline, oxiracetam and the like.

Further, the present invention provides a pharmaceutical or nutraceutical composition for preventing and treating a brain nerve damage disease, the pharmaceutical or health care composition comprising a ginkgolide synergist and a treatment for a brain nerve damage disease or A pharmaceutically active molecule that acts as a health care.

Further, the ginkgolide synergist is at least one of a natural product of ginkgolide, a lactone lactone extract, a natural product ginkgolide or a hydroxy derivative of a ginkgolide extract;

Further, the ginkgolide extract is at least one of a ginkgolide extract or a ginkgo biloba extract, and the ginkgolide extract has a ginkgolide content of from 1 to 99%.

Further, the hydroxy derivative of the natural product ginkgolide or biloba lactone extract is a glycoside, ester or ether thereof.

Further, the medicinal active molecule having therapeutic or health-care effects on brain nerve injury diseases is selected from the group consisting of ginseng total saponins, shouwu stilbene glycosides, resveratrol, levodopa, edaravone, and vinca At least one of statin, nigeroline, citicoline or oxiracetam.

Further, the ginkgolide, the biloba lactone extract, and the hydroxy derivative thereof, particularly the glycoside, ester, and ether derivative thereof, can be used as a medicine having therapeutic or health-care effects on brain nerve damage diseases. The active ingredients are used in combination in the form of a health supplement.

The inventors have conducted a large number of research experiments, showing that biloba lactones, especially bilobalide, ginkgolides and their hydroxy derivatives, especially their glycosides, esters, ether derivatives and ginsenosides, Shouwu diphenyl Promote these active ingredients when combined with natural medicinal active ingredients such as ethylene glycoside and resveratrol or chemical components such as levodopa, edaravone, vinpocetine, ergoline, citicoline, and oxiracetam. Through the blood-brain barrier, the composition of ginsenosides, stilbene glucoside, resveratrol, levodopa, edaravone, vinpocetine, ergoline, citicoline, oligo The concentration of rasacetine in brain tissue, and promotes neural stem cell proliferation and directed differentiation, anti-cerebral ischemic injury, neuroprotection, promoted differentiation of neural stem cells, improved learning and memory, and anti-senile dementia, and the main active ingredients are clear The quality is stable, and the side effects of the active ingredient in the peripheral tissues are reduced, thereby reducing the toxicity and increasing the effect.

When the medicinal component of the brain nerve damage disease is ginseng total saponin and shouwu stilbene glycoside, the weight ratio of ginseng lactone synergist: ginseng total saponin: Shouwu stilbene glycoside is 1: 10 to 60: 10 to 50.

When the medicinal active ingredient for preventing and treating brain nerve damage diseases is ginseng total saponin and resveratrol, the weight ratio of biloba lactone synergist: ginseng total saponin: resveratrol is 1:10-25: 10 to 20.

When the pharmaceutically active ingredient for preventing and treating brain nerve damage diseases is vinpocetine, the weight ratio of the biloba lactone synergist and vinpocetine is 1:10-25.

When the medicinal component of the brain nerve damage disease is levodopa, the weight ratio of the biloba lactone synergist and levodopa is 1:10 to 50;

When the pharmaceutical active ingredient for preventing and treating brain nerve damage disease is nigeroline, the weight ratio of ginkgolide synergist and nigeroline is 1:10-50;

When the medicinal component of the brain nerve damage disease is oxiracetam, the weight ratio of the biloba lactone synergist and oxiracetam is 1:20-50;

When the medicinal component of the brain nerve damage disease is edaravone, the weight ratio of the biloba lactone synergist and edaravone is 1:20-50;

When the active ingredient of the brain nerve damage disease is citicoline, the weight ratio of the lactone lactone synergist and the citicoline is 1:20-50.

The brain nerve injury disease according to the present invention is a cerebral ischemic injury disease or a neurodegenerative disease, wherein the cerebral ischemic injury disease is ischemic stroke, chronic cerebral ischemia, stroke sequelae or vascular dementia. The neurodegenerative disease is senile dementia, Parkinson's disease, Alzheimer's disease or mild cognitive impairment.

The advantages of the present invention over the prior art are:

The inventors have unexpectedly discovered that ginkgolide compounds, particularly ginkgolide, ginkgolide extract (ginkgolide extract or ginkgo biloba extract), and its hydroxy derivatives, especially its glycosides, have been extensively tested. , esters, ether derivatives, can promote the treatment or health care of brain nerve damage diseases, such as ginseng total saponins, Shouwu stilbene or resveratrol, or other natural medicinal active ingredients or Levodopa, edaravone, vinpocetine, nigralin, citicoline, oxiracetam and other chemical components pass through the blood-brain barrier and enter the brain tissue without increasing the blood drug concentration of the drug molecule. The concentration of drug molecules in brain tissue is greatly increased, and the effects of promoting neural stem cell proliferation and directed differentiation, anti-cerebral ischemic injury, neuroprotection, promoting differentiation of neural stem cells, improving learning and memory, and resisting senile dementia are effective. The efficacy of the drug has been improved and unexpected technical effects have been achieved.

In addition, the pharmaceutical composition for preventing and treating brain nerve damage diseases provided by the invention has the main active ingredients and stable quality, and reduces the side effects of the active ingredients in the peripheral tissues by promoting the entry of the active drug molecules into the brain tissue, thereby reducing toxicity and increasing efficiency. effect. At the same time, the use of a small dose can be made into a variety of sustained release controlled release preparations and large-scale production. Further, the ginkgolide compound used in the present invention has a wide range of sources, is low in production cost, and is easy to use widely.

DRAWINGS

Figure 1 shows the reduction of cell resistance in the BBB model by ginkgolide. Among them, Control: normal control group, Bilo: bilobalide group, SCH: A2a adenosine receptor inhibitor group, SCH+: A2a adenosine receptor inhibitor + bilobalide group, DPCPX: A1 adenosine receptor inhibitor Group, DPCPX+: A1 adenosine receptor inhibitor group + bilobalide group, compared with the normal group, ^** P <0.01; ^*** P <0.001.

Figure 2 is a graph showing the effect of ginkgolide on the penetration of sodium fluorescein into the microvascular endothelium of brain tissue. Among them, Control: normal control group, Bilo: bilobalide group, DPCPX: A1 adenosine receptor inhibitor group, DPCPX+: A1 adenosine receptor inhibitor group + bilobalide group, compared with normal group, ^*** P < 0.001.

Figure 3 is a graph showing the effect of bilobalide on the expression of BBB microvascular endothelial adenosine receptor. Among them, Control: normal control group, Bilo L: low dose of bilobalide, Bilo H: high dose of bilobalide, Egb L: low dose of Ginkgo biloba extract, Egb H: high dose of Ginkgo biloba extract, CCPA: adenosine A1 receptor agonist (2-chlorocyclopentadenosine) group, Lexican: A2a adenosine receptor agonist group, NECA: non-selective adenosine receptor agonist (5-N-ethyl amide group) Adenosine group; ^* P<0.05; ^** P<0.01 compared with normal group.

Figure 4 is a graph showing the inhibition of BBB microvascular endothelial ZO-protein receptor expression by ginkgolide. Among them, Control: normal control group, DPCPX+: A1 adenosine receptor inhibitor group, DPCPX+Bilo: A1 adenosine receptor inhibitor + bilobalide group, DPCPX+Egb: A1 adenosine receptor inhibitor + ginkgo leaf Extract group.

detailed description

The invention is further described below by way of specific embodiments, but the invention is not limited to the following examples.

The ginkgolide and the biloba lactone extract according to the present invention can be obtained by purchasing a commercially available product, or can be isolated and purified by the existing method, wherein the preparation and identification method of the ginkgolide can be referred to the China with the publication number CN101829109B. The patent "the new use of albino lactone and its derivatives with aquaporin regulation in the treatment of brain edema" is disclosed by using a reflux extraction combined with column chromatography separation method to obtain ginkgolide from the ginkgo leaf powder, and The purity of bilobalide was determined by HPLC technique, and the structure of bilobalide was confirmed by IR, MS and NMR techniques. In addition, the preparation method of ginkgo lactone can also be referred to from the Ginkgo biloba leaf by high-speed countercurrent chromatography (HSCCC) disclosed in Chinese Patent Application No. CN102670586A, "Application of Ginkgolide in Preparation of Medicine for Treating Ischemic Cardiomyopathy" The bilobalide monomer was isolated and prepared from the crude extract and identified by TLC.

Example 1

Open study on the blood-brain barrier (in vitro BBB model) of bilobalide and its extracts:

1. Experimental plan:

1.1 Establishment of an in vitro blood-brain barrier model (BBB): cultured human glial cells (HEB cells) and immortalized brain microvascular endothelial cells (hCMEC/D3 cells). After the cells are full, they are digested and centrifuged to HEB cells. The density of 3╳10 ⁵ /mL was inoculated into the lower layer of the transwell 12-well plate chamber. After the cells were attached, the cells were cultured in low-sugar medium. When the cells grew to 80%, the upper layer of the transwell chamber was 7╳10 ⁵ /mL. The density of the hCMEC/D3 cells was inoculated. After the cells were overgrown, the trans-endothelial resistance (TEER) of the co-culture model was determined by Millicell ERS as the sum of endothelial cells (EC) and TEER, in a cell-free culture cell. As a control, the resistance value of the TEER was measured, and the resistance value of the single-layer BCEC was: TEEREC=(TEEREC+filter-TEERfilter)╳S (membrane area), and the unit was Ω·cm ² . When the lateral cell resistance is greater than 300 Ω·cm ² , the BBB in vitro model is basically established;

1.2 Determination of sodium fluorescein permeability and related protein expression:

10 mg/L sodium fluorescein was administered to the upper layer of the transwell chamber. After 1 h, the medium in the lower chamber was aspirated, and the permeability of sodium fluorescein was measured by a microplate reader. When the permeability was 1 to 30 ╳ 10 ^-6 cm / The range of s represents the success of the in vitro BBB model. Give ginkgolide (extract) or its derivative, determine the resistance of the drug to the BBB model in vitro and the transmittance of sodium fluorescein in different time periods, determine the efficacy of the drug and the time of drug action It is 2h.

Set normal group, bilobalide (extract) or its derivatives in low, medium and high dose groups (0.2,1,5μg/mL), A1 adenosine receptor agonist CCPA (1μmol), A2a adenosine receptor The agonist Lexican (1 μmol) and the broad-spectrum agonist NECA (1 μmol) were measured for their resistance and sodium fluorescein transmission after 2 hours of administration. By administering the A1 adenosine receptor inhibitor DPCPX (50 μmol), the A2a adenosine receptor inhibitor SCH (50 μmol), and simultaneously setting the inhibitor + bilobalide (extract) or its derivative (1 μg/mL), The change in resistance and the transmittance of sodium fluorescein were measured, and the target was verified in the reverse direction. The results are shown in Figures 1 and 2. At the same time, the proteins of each group were extracted, and the expression of adenosine receptor and tight junction-associated protein was detected by Western Blot to verify whether it regulates the expression of adenosine receptors and regulates the expression of tight junction-associated proteins, thereby increasing the blood-brain barrier. The permeability is shown in Figures 3 and 4.

2. Experimental results:

As can be seen from Fig. 1, ginkgolide and its hydroxy derivatives, especially its glycosides, esters, ether derivatives, etc., can reduce the electrical resistance of BBB cells, increase the permeability of BBB, and promote the entry of drug molecules into brain tissue.

As can be seen from Fig. 2, compared with the normal group, bilobalide significantly increased the osmotic concentration of sodium fluorescein by 70%; the osmolality of sodium fluorescein was also significantly increased after administration of the A2a adenosine receptor inhibitor SCH (P< 0.05).

3 and 4, ginkgolide and its hydroxy derivatives, especially its glycosides, esters, ether derivatives, etc., can promote the expression of adenosine receptor (AR) in the microvascular endothelium of brain tissue and inhibit tight junction protein ZO. -1 protein expression, improve BBB permeability, and promote the entry of drug molecules into brain tissue.

Example 2

Ginkgolide extract (glycolide content ≥ 99.0%) 5, 15g;

Active ingredient of traditional Chinese medicine: ginsenoside extract (ginsenoside Rg1 content ≥ 95%) 300g;

The above-mentioned various extracts were weighed and mixed uniformly in an equal amount, and the obtained mixture was recorded as YXN1-1 (containing 5 g of ginkgolide extract and 300 g of ginsenoside extract) and YXN1-2 (containing 15 g of ginkgo). Ester extract and 300 g of ginsenoside extract).

Pharmacodynamic experiment:

Treatment group: YXN1-1 and YXN1-2 were made into a suspension, and the YXN1-1 treatment group was dosed at 101.6 mg/kg (equivalent to ginsenoside extract 100 mg/kg), 20 mL/kg capacity, and 5 SD were administered. Rats; YXN1-2 treatment group was administered with 105 SD/kg dose (corresponding to ginsenoside extract 100 mg/kg), 20 mL/kg capacity, and 5 SD rats were intragastrically administered;

Control group: another ginsenoside extract was prepared as a suspension. According to the ginsenoside extract 100 mg/kg dose, 20 mL/kg capacity, 5 SD rats were intragastrically administered;

After 0.5 h of administration, blood was taken, the rats were acutely sacrificed, brain tissue was taken, and brain homogenate was prepared. UPLC-MS was used to detect the content of ginsenoside Rg1 in blood and brain tissue.

Another 40 MCAO forebrain ischemia was prepared and divided into 4 groups and 10 groups/group. The vehicle saline, YXN1-1, YXN1-2 and ginsenoside extract were administered respectively. The administration method and dosage were the same as above. After 7 days of administration, The animals were sacrificed and the improvement rate of cerebral infarct size was observed.

The results are shown in Table 1:

Table 1 UPLC-MS content determination of YXN1 and results of promoting BBB

The experimental results showed that the concentration of ginsenoside Rg1 in the brain tissue of YXN1 was 5.4/9.4 times higher than that of ginsenoside-administered rats, when YXN1 and ginsenoside extracts containing the same amount of Rg1 were administered. Indicates ginkgolide and ginsenoside Rg1 In combination, it can promote the ginsenoside Rg1 to penetrate the blood-brain barrier and improve the protective effect of cerebral ischemia.

Example 3

Ginkgolide extract (glycolide content ≥ 20.0%) 20, 40g;

Active ingredient of traditional Chinese medicine: ginsenoside extract (ginsenoside Rg1 content ≥ 30%) 200g;

The above-mentioned various extracts were weighed and mixed uniformly in an equal amount, and the obtained mixture was recorded as YXN2-1 (containing 20 g of ginkgolide extract and 200 g of ginsenoside extract) and YXN2-2 (containing 40 g of ginkgo). Ester extract and 200 g of ginsenoside extract).

Pharmacodynamic experiment:

Treatment group: YXN2-1 and YXN2-2 were made into suspension. The YXN2-1 treatment group was dosed at 220 mg/kg (equivalent to ginsenoside extract 200 mg/kg), 20 mL/kg capacity, and 5 SD large. Rats; YXN2-2 treatment group according to the dose of 240mg / kg (equivalent to ginsenoside extract 200mg / kg), 20mL / kg capacity, gavage 5 SD rats;

Control group: another ginsenoside extract was prepared as a suspension. According to the ginsenoside extract 200mg/kg dose, 20mL/kg capacity, 5 SD rats were intragastrically administered;

After 0.5 h of administration, blood was taken, the rats were acutely sacrificed, brain tissue was taken, and brain homogenate was prepared. UPLC-MS was used to detect the content of ginsenoside Rg1 in blood and brain tissue.

Another 40 MCAO forebrain ischemia was prepared and divided into 4 groups and 10 groups/group. The vehicle saline, YXN2-1, YXN2-2 and ginsenoside extract were administered respectively. The administration method and dosage were the same as above. After 7 days of administration, The animals were sacrificed and the improvement rate of cerebral infarct size was observed.

The results are shown in Table 2:

Table 2 Determination of UPLC-MS content of YXN2 and results of promoting BBB

The experimental results showed that the concentration of ginsenoside Rg1 in the brain tissue of YXN2 was 6.5/10 higher than that of ginsenoside extract alone, when YXN2 and ginsenoside extracts containing equal amounts of Rg1 were administered. Times. It indicates that the combination of ginsenoside and ginsenoside Rg1 can promote the ginsenoside Rg1 to penetrate the blood-brain barrier and improve the protective effect of cerebral ischemia.

Example 4

Ginkgolide extract (glycolide content ≥ 95.0%) 5, 10g;

Active ingredient of traditional Chinese medicine: Polygonum multiflorum extract (Shouwu stilbene EB content ≥ 50%) 200g;

The above-mentioned various extracts were weighed and mixed uniformly in an equal amount, and the obtained mixture was recorded as YXN3-1 (containing 5 g of ginkgolide extract and 200 g of Polygonum multiflorum extract) and YXN3-2 (containing 10 g of ginkgolide). Extract and 200g of Polygonum multiflorum extract).

Pharmacodynamic experiment:

Treatment group: YXN3-1 and YXN3-2 were prepared as suspension. The YXN3-1 treatment group was dosed at 102.5 mg/kg (equivalent to 100 mg/kg of Polygonum multiflorum extract), 20 mL/kg capacity, and 5 SD large. Rats; YXN3-2 treatment group according to the dose of 105mg / kg (equivalent to Polygonum multiflorum extract 100mg / kg), 20mL / kg capacity, gavage 5 SD rats;

Control group: Another extract of Polygonum multiflorum was prepared as a suspension. According to the dose of 100 mg/kg of Polygonum multiflorum extract, 20 mL/kg capacity, 5 SD rats were intragastrically administered;

After 0.5 h of administration, the blood was taken, the rats were acutely sacrificed, brain tissue was taken, and brain homogenate was prepared. UPLC-MS was used to detect the content of Shouwustilbene in blood and brain tissue.

Another 40 cases of 4-VO cerebral ischemia were prepared and divided into 4 groups, 10 rats/group, respectively, and the vehicle saline, YXN3-1, YXN3-2 and Polygonum multiflorum extract were administered. The administration method and dosage were the same as above. Animals were sacrificed in the day to observe the rate of nerve regeneration and learning and memory improvement.

The results are shown in Table 3:

Table 3 UPLC-MS content determination of YXN3 and results of promoting BBB

The experimental results showed that the blood concentration was not significantly different with YXN3 and Polygonum multiflorum extracts containing the same amount of S. sylvestrein EB, but the EB content in the brain tissue of YXN3 was 4.6 higher than that of the rats fed with Polygonum multiflorum extract. /9 times. It indicates that the combination of bilobalide and Shouwu stilbene glucoside can promote the penetration of Shouwu stilbene glycoside through the blood-brain barrier, have brain-targeting effect, and can improve the anti-cerebral ischemic injury and improve learning of stilbene glycoside. Memory effect.

Example 5

Ginkgolide extract (glycolide content ≥ 98.0%) 5g;

Resveratrol (resin content of 99%) 100g;

The above ingredients were weighed and mixed uniformly in an equal amount, and the obtained mixture was designated as YXN4.

Pharmacodynamic experiment:

Treatment group: YXN4 was made into a suspension, and 5 SD rats were intragastrically administered at a dose of 105 mg/kg (corresponding to 100 mg/kg of resveratrol) and a volume of 20 mL/kg.

Control group: another suspension of resveratrol was prepared, and 5 SD rats were intragastrically administered with a dose of resveratrol at a dose of 100 mg/kg and a volume of 20 mL/kg.

After 0.5 h of administration, the blood was taken, the rats were acutely sacrificed, brain tissue was taken, and brain homogenate was prepared. UPLC-MS was used to detect the content of resveratrol in blood and brain tissue.

Another 30 MCAO forebrain ischemia was prepared and divided into 3 groups, 10 rats/group, respectively. The vehicle was treated with normal saline, YXN4, and resveratrol. The administration method and dosage were the same. After 7 days of administration, the animals were sacrificed and the brain was observed. Infarct area improvement rate.

The results are shown in Table 4:

Table 4 Determination of UPLC content of YXN4 and results of promoting BBB

The experimental results showed that the blood concentration of YXN4 and resveratrol alone was not significant, but the content of resveratrol in the brain tissue of YXN4 was higher than that of rats treated with resveratrol alone. At 8.1 times higher, the improvement rate of cerebral infarct size in MCAO cerebral ischemia rats was increased by 1.54 times. It shows that the combination of ginkgolide derivatives and resveratrol can promote the respiration of resveratrol through the blood-brain barrier, have a brain-targeting effect, and can improve the anti-cerebral ischemic injury.

Example 6

YXN5: commercially available ginkgolide extract (content ≥ 20%), ginseng total saponin extract (content ≥ 30%), Shouwu stilbene glycoside (content ≥ 50%), according to the weight of 1:12:8 composition.

YXN6: commercially available ginkgolide (content ≥ 60%), ginseng total saponin extract (content ≥ 90%), Shouwu stilbene glycoside (content ≥ 60%), composed of 1:10:5 by weight .

RSSW: ginseng total saponin extract (content ≥ 90%), Shouwu stilbene glycoside (content ≥ 60%), composed of 10:5 parts by weight.

Pharmacodynamic experiment:

Treatment group: YXN5, YXN6, RSSS were made into suspension, and 5 SD rats were intragastrically administered at a dose of 100 mg/kg and a volume of 20 mL/kg.

Control group: Another ginseng total saponin extract (content ≥ 90%) was prepared as a suspension, and 5 SD rats were intragastrically administered at a dose of 50 mg/kg and a volume of 20 mL/kg.

0.5 h after administration, blood was taken, the rats were acutely sacrificed, brain tissue was taken, and brain homogenate was prepared. UPLC-MS was used to detect the content of active ingredients in blood and brain tissue.

Another 50 MCAO forebrain ischemia was prepared and divided into 5 groups and 10 groups/group. The vehicle was treated with normal saline, YXN5, YXN6, RSSW and total ginsenoside extract. The administration method and dosage were the same as above. After 7 days of administration, the drug was sacrificed. Animals, observe the improvement rate of cerebral infarction area, observe the rate of nerve regeneration and learning and memory improvement.

The results are shown in Table 5-7:

Table 5 Active ingredient content in blood (μg/mL) at 0.5 h of different compositions

Test items Ginsenoside saponins RSSW YXN5 YXN6 Stilbene 0 48.5±6.3 26.9±3.56 26.1±3.81 Ginsenoside Rg1 25.1±3.12 25.2±3.32 25.7±3.36 25.9 ± 3.69 Ginsenoside Rd 4.8±0.63 4.7±0.65 4.5±0.71 4.3±0.68 Ginsenoside Rb1 21.1±2.89 21.3±3.12 21.1±3.08 21.9±3.16 Ginsenoside Re 4.2±0.59 4.4±0.61 4.6±0.63 4.3±0.59 Ginkgolide 0 0 1.05±0.21 3.75±0.46

Table 6 Active ingredient content in brain tissue after 0.5 h administration of different compositions (ng/mL)

Test items Ginsenoside saponins RSSW YXN5 YXN6 Stilbene 0 4.9±0.62 38.1±4.32 ^** 134.2±16.5 ^** Ginsenoside Rg1 2.24±0.31 10.1±1.32 93.5±12.3 ^** 131.5±13.2 ^** Ginsenoside Rd 0.46±0.06 4.6±0.58 25.4±4.21 ^** 31.6±4.63 ^** Ginsenoside Rb1 2.21±0.32 4.5±0.58 118.2±11.6 ^** 123.9±13.1 ^** Ginkgolide 0 0 9.8±1.23 18.4±2.21

Note: ^** P < 0.01 compared with RSSS without the addition of ginkgolide extract brain targeting agent.

It can be seen from Table 5-6 that the different concentrations of ginseng total saponins and Shouwustilbene glycosides are not significantly different, but after combination with biloba lactone extracts, ginseng in brain tissue The content of total saponins and Shouwu stilbene glycosides is increased by 5 to 40 times.

Table 7 Protective effects of cerebral ischemia after 7 days of administration of different compositions

Test items Ginsenoside saponins RSSW YXN5 YXN6 Infarct size (%) 39.13±5.32 34.46±5.62 23.5±4.28 ^** 21.09±3.98 ^** Nerve regeneration rate (%) 86.8±9.85 91.6±10.21 191.8±21.3 ^** 282.6±23.9 ^** Learning and memory improvement rate 19.91±2.61 24.81±3.89 73.6 ± 8.67 ^** 77.41±9.13 ^** Space exploration distance (cm) 25.21±4.32 45.23±5.79 74.82±8.91 ^** 80.19±9.93 ^**

Note: ^** P < 0.01 compared with RSSS without the addition of ginkgolide extract brain targeting agent.

As can be seen from Table 7, 7 days after the administration of YXN5 and YXN6, the area of cerebral infarction can be reduced, the rate of brain nerve regeneration can be improved, and the improvement, improvement of learning and memory, and spatial exploration distance (cm) can be prolonged. The potency is stronger than the ginseng total saponin extract or RSSW alone. Ginkgolide extract promotes the penetration of active ingredients of traditional Chinese medicine through the blood-brain barrier, enhances anti-cerebral ischemic injury and neuroprotection, and improves learning and memory.

Example 7

YXN7: commercially available ginkgolide extract (content ≥ 60%), vinpocetine (content ≥ 98%), composed of 10:25 parts by weight, made into tablets 5mg / tablet;

YXN8: commercially available ginkgolide (content ≥98%), edaravone (content ≥98%), composed of 10:30 parts by weight;

YXN9: commercially available ginkgolide (content ≥98%), oxiracetam (content ≥98%), composed of 10:35 parts by weight, made into capsules, 400 mg / granule.

Pharmacodynamic experiment:

Treatment group: YXN7, YXN8, YXN9 were separately prepared as suspension, YXN7 was administered with 5 SD rats at a dose of 5 mg/kg and 20 mL/kg; YXN8 was administered at a dose of 15 mg/kg and 20 mL/kg for 5 SD large Rats; YXN9 was administered with 5 SD rats at a dose of 400 mg/kg and a volume of 20 mL/kg.

Control group: Another suspension of vinpocetine, edaravone, and oxiracetam was prepared at a dose of 5 mg/kg, 15 mg/kg, and 400 mg/kg, and a volume of 20 mL/kg was administered to 5 SD large groups. mouse.

0.5 h after administration, blood was taken, the rats were acutely sacrificed, brain tissue was taken, and brain homogenate was prepared. UPLC-MS was used to detect the content of active ingredients in blood and brain tissue.

Another 70 MCAO forebrain ischemia rats were prepared and divided into 7 groups, 10 rats in each group, respectively, given vehicle physiological saline, YXN7, YXN8, YXN9, vinpocetine, edaravone, oxiracetam, and administered. The method and dose were the same as above. After 7 days of administration, the animals were sacrificed to observe the improvement rate of cerebral infarct area, and the rate of nerve regeneration and learning and memory improvement was observed.

The results are shown in Table 8-10:

Table 8 Active ingredient content in blood (μg/mL) at 0.5 h after administration of different compositions

Group Vinpocetine YXN7 Edaravone YXN8 Olaxitan YXN9 Vinpocetine 8.23±1.32 7.94±1.26 / / / / Edaravone / / 5.31±0.65 5.19±0.63 / / Olaxitan / / / / 82.32±9.68 84.52±10.21

Table 9 Active ingredient content in brain tissue after 0.5 h administration of different compositions (ng/mL)

Group Vinpocetine YXN7 Edaravone YXN8 Olaxitan YXN9 Vinpocetine 12.6±1.85 36.1 ± 4.61 ^** / / / / Edaravone / / 122.5±13.2 411.2±54.2 ^** / / Olaxitan / / / / 182.8±19.6 961.3±102.8 ^**

Table 10 Protective effects of cerebral ischemia after 7 days of administration of different compositions

Note: ^** P < 0.01 compared with the corresponding drug without the addition of the ginkgolide extract brain targeting agent.

It can be seen from Table 8-9 that different concentrations of vinpocetine, edaravone, and oxiracetam are administered, and the difference in blood concentration between the two is shown. Not significant, but combined with the biloba lactone extract, the content of vinpocetine, edaravone, and oxiracetam in brain tissue increased by 3 to 4 times.

As can be seen from Table 10, 7 days after administration of YXN7, YXN8 and YXN9, the infarct size can be reduced, the rate of brain nerve regeneration can be improved, and the improvement, the improvement of learning and memory, and the space exploration distance (cm) can be prolonged. The efficacy is stronger than that of vinpocetine, edaravone, and oxiracetam. Ginkgolide extract promotes the penetration of active ingredients of traditional Chinese medicine through the blood-brain barrier, enhances anti-cerebral ischemic injury and neuroprotection, and improves learning and memory.

Example 8

YXN10: commercially available ginkgolide extract (content ≥ 60%), levodopa, composed of 10:40 parts by weight;

YXN11: commercially available ginkgolide (content ≥ 98%), levodopa, consisting of 10:50 parts by weight.

Pharmacodynamic experiment:

Treatment group: YXN10 and YXN11 were prepared as suspensions, and 5 SD rats were intragastrically administered with a dose of 150 mg/kg of levodopa and a volume of 20 mL/kg.

Control group: Another suspension of levodopa was prepared, and 5 SD rats were intragastrically administered at a dose of 150 mg/kg and a volume of 20 mL/kg.

0.5 h after administration, blood was taken, the rats were acutely sacrificed, brain tissue was taken, and brain homogenate was prepared. UPLC-MS was used to detect the content of active ingredients in blood and brain tissue.

Another 40 rats of 6-hydroxydopamine Parkinson's disease model were prepared and divided into 4 groups, 10 rats in each group, respectively, given vehicle physiological saline, YXN10, YXN11 and levodopa. The administration method and dosage were the same as above. In the day, the changes of the five indicators such as the slow-moving experiment, the grasping experiment, the tail stiffness, the change of the tonic symptoms, and the tremor experiment were observed.

The experimental results are shown in Table 11-12:

Table 11 levodopa content in blood and brain tissue at 0.5 h after administration of different compositions

Group Levodopa YXN10 YXN11 Blood content (μg/mL) 102.3±13.6 101.5±12.2 99.7±11.8 Brain tissue content (ng/mL) 281.6±31.7 878.3±103.1 ^** 891.5±110.6 ^**

Note: ^** P < 0.01 compared with the levodopa group without the addition of the ginkgolide extract brain-targeting agent.

Table 12 Protection of 6-hydroxydopamine Parkinson's disease after 15 days of administration of different compositions

Test items Model group Levodopa YXN10 YXN11 Slow action duration (min) 52.5±7.6 18.5±2.5 ^## 11.5±2.1 ^** 9.6±2.1 ^** Grasping experiment duration (min) 31.8±4.1 52.1±6.8 ^## 65.6 ± 7.2 ^** 64.8 ± 7.5 ^** Tail strong time (min) 36.1 ± 4.6 11.6±1.8 ^## 7.7±2.5 ^** 6.2 ± 1.9 ^** Muscle tremor frequency (times / min) 54.2±6.8 130.1±12.6 ^## 8.9±2.3 ^** 8.6±2.6 ^** EMG group discharge frequency (times / s) 7.7±2.1 3.4±1.2 ^## 2.9±1.3 ^** 2.8±1.5 ^**

Note: Compared with the model group, ^## P<0.01; compared with the levodopa group without the addition of the ginkgolide extract brain target agent, ^** P < 0.01.

As can be seen from Table 11, the difference in blood drug concentration was not significant when different compositions containing equal amounts of levodopa were administered, but after combination with the ginkgolide extract, the content of levodopa in the brain tissue was increased by three times.

As can be seen from Table 12, 15 days after the administration of YXN10 and YXN11, the rats' movement retardation test, grasping experiment and tail can be significantly improved. Symptoms such as tonicity, significantly reduce the frequency of muscle tremor and the frequency of electromyogram group discharge. The efficacy is stronger than levodopa alone. Ginkgolide extract promotes levodopa through the blood-brain barrier and enhances its pharmacological effect on Parkinson's disease.

Test Example 1. Application in clinical treatment of ischemic stroke sequelae

1. Clinical data and methods

1.1 General Information

Sixty volunteers with cerebral infarction sequelae were screened and randomly divided into ginseng total saponin capsules, RSSW capsules, and YXN5 capsule treatment groups, 20 in each group. Among the 20 patients with ginseng total saponin capsule group, 13 were male and 7 were female; the age ranged from 38 to 72 (51.6±6.6) years; the course of disease was 37-82 (51.0±7.1) days. Among the 20 patients in the RSSW capsule group, 13 were male and 7 were female; the age ranged from 37 to 74 (51.8±7.0) years; the course of disease was 37-81 (51.8±7.3) days. Among the 20 patients in the YXN5 capsule control group, 13 were male and 7 were female; the age ranged from 37 to 74 (52.0±7.1) years; the course of disease was 38-83 (52.6±7.9) days. There were no significant differences in the demographic data of age, gender, disease duration, body weight, health history, medication history, illness history, and stroke type among the three groups (P>0.05), which were comparable.

1.2 inclusion criteria

(1) The diagnosis of Western medicine is in accordance with the "Diagnostic Points for Various Cerebrovascular Diseases" revised by the Fourth Annual Conference of Cerebrovascular Diseases of the Chinese Medical Association in 1995; and confirmed by CT or MRI; (2) Symptomatic treatment of acute phase Relief, no other serious comorbidities; (3) TCM diagnosis is in line with the "Standards for Diagnostic Efficacy of Stroke Diseases" (Trial) standards drafted by the State Administration of Traditional Chinese Medicine Administration's Brain Disease Emergency Research Collaboration Group; (4) Ages 40 to 80 years old; (5) The stage of the disease is the recovery period (2 weeks to 6 months); (6) The sequelae such as left hemiplegia, numbness of the limbs, skewed eyes and mouth, and unfavorable language; (7) Patients voluntarily sign informed consent.

1.3 exclusion criteria

(1) patients with acute stroke; transient ischemic attack, cerebral hemorrhage or asymptomatic cerebral infarction; (2) severe joint deformity affects functional recovery; (3) complicated with severe heart, liver and kidney dysfunction; (4) patients are unwilling to cooperate; (5) pregnant or lactating women, allergic or allergic to known components of the drug; (6) unstable vital signs, need emergency guardian; (7) mental, mentally handicapped (8) Patients with secondary stroke induced by other causes such as tumors and blood diseases.

1.4 treatment methods

Ginsenoside saponin capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

RSSW capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

YXN5 capsule group: 125 ~ 250mg, 3 times / d, according to the condition, individualized administration.

The composition of the components in each group of capsules is shown in Example 6.

Three groups of patients were given ginseng total saponin capsules, RSSW capsules and YXN5 capsules. The treatment of each group was started after the treatment, and the treatment was continued for 14 days. The observation time was 4 courses.

1.5 observation indicators

Observation indicators were used to record the scores of neurological deficits and activities of daily living activities before and after treatment in the two groups.

The curative effect criteria were cured: after treatment, the clinical symptoms and signs disappeared or disappeared, the treatment index was 95%; markedly effective: the signs after treatment were significantly improved, 70% ≤ treatment index <95%; effective: signs after treatment improved, 30% ≤ treatment index <70%; invalid: no improvement in signs after treatment, treatment index <30%.

Total effective rate = (healing + effective + effective) / total number of cases

表示，组间比较采用t检验。均以P＜0.05为差异有统计学意义。1.6 statistical methods using SPSS 19.0 software package for data processing, counting data comparison using χ ² test, measurement data used

It is indicated that the t-test is used for comparison between groups. The difference was statistically significant at P<0.05.

2. Therapeutic results

2.1 Neurological deficit score (NIHSS)

The total effective rate of the three groups was higher, and the neurological function was significantly improved after treatment. Compared with the three groups, the effect of ginseng total saponin capsule <RSSW capsule <YXN5 capsule, no significant difference, see Table 13.

Table 13 Comparison of neurological deficit scores (NIHSS) in each group

Group Number of cases Before treatment After treatment YXN5 capsule 20 17.1±5.6 7.3±4.1 ^**# RSSW capsule 20 17.2±5.7 10.6±4.3 ^* Ginseng total saponin capsule 20 16.9±5.9 13.3±2.9 ^*

Note: compared with before treatment in each ^{group, * P <0.05, ** P} <0.01; compared with total ginsenoside capsule group, ^# P <0.05.

2.2 Assessment of daily living ability (BL)

The ability of daily life after treatment was significantly improved in the three groups. Compared with the control group, the effect of brain-targeted therapy was better than that of the control group. The difference was statistically significant (see Table 14).

Table 14 Comparison of daily life ability status (BL) scores before and after treatment in each group of patients

Group Number of cases Before treatment After treatment YXN5 capsule 20 53.9±19.1 91.5±15.1 ^**# RSSW capsule 20 54.1±20.6 85.6±14.2 ^* Ginseng total saponin capsule 20 53.2±21.9 78.1±12.3 ^*

Note: compared with before treatment in each ^{group, * P <0.05, ** P} <0.01; compared with total ginsenoside capsule group, ^# P <0.05.

2.3 stroke efficacy

The total effective rate of the three groups was higher, and the curative effect was obvious after treatment. The efficacy of YXN5 capsule group was 90%, which was better than RSSW capsule and ginseng total saponin capsule group. The difference was statistically significant (chi-square test, ^* P<0.05). See Table 15.

Table 15 Comparison of the efficacy of stroke in each group

Group Total number of cases get well Significant effect effective invalid Total efficiency /% YXN5 capsule 20 6 8 4 2 90 ^* RSSW capsule 20 4 7 5 4 80 Ginseng total saponin capsule 20 2 6 6 6 70

The above results showed that the efficacy of YXN5 containing bairuactone brain-targeted potentiator in the treatment of stroke and stroke sequelae was significantly improved (P<0.05).

Test Example 2: Application in clinical treatment of Parkinson's disease

1. Clinical data and methods

1.1 General Information

All cases from 2010 to 2014 were selected from 100 patients with neurological subjects who met the diagnostic criteria of Parkinson's disease and were diagnosed with Parkinson's disease by clinical examination. They were randomly divided into ginseng total saponin capsule group, RSSW capsule group, YXN5 capsule group, YXN10 capsule group and levodopa group. Among them, there were 20 cases of ginseng total saponin capsule group, 12 males and 8 females; the course of disease was 1-12 years. , an average of 3.7 years; aged 47 to 70 years, an average of 56.4 years. There were 20 patients in the RSSW capsule group, 12 males and 8 females; the course of disease was 1 to 12 years, with an average of 3.7 years; the age ranged from 48 to 70 years, with an average of 56.2 years. There were 20 cases in the YXN5 capsule group, 12 males and 8 females; the course of disease was 1 to 12 years, with an average of 3.8 years; the age ranged from 49 to 70 years, with an average of 57.1 years. There were 20 cases in the YXN10 capsule group, 11 males and 9 females; the course of disease was 1 to 13 years, with an average of 3.8 years; the age ranged from 50 to 75 years, with an average of 62.1 years. There were 20 patients in the levodopa tablet group, 11 males and 9 females; the course of disease was 1 to 12 years, with an average of 3.8 years; the age ranged from 51 to 76 years, with an average of 62.5 years. The 5 groups were statistically treated in terms of gender, disease duration and age. The differences were not significant (P>0.05) and were comparable.

Among them, the diagnostic criteria: diagnosis based on the 2010 approved Parkinson's disease diagnosis and treatment guidelines (Beijing Union Medical College Hospital. Parkinson's disease diagnosis and treatment guidelines. Chinese clinicians, 2010, 38 (2): 77-79.) Sexual PD patients.

Exclusion criteria: (a) Parkinson's disease caused by severe heart, lung and kidney dysfunction, secondary to cerebrovascular disease, trauma and other neurological and psychiatric disorders; (b) PD superimposed syndrome; (c) suffering from Malignant tumors, disability, and other serious cases of severe neurological, hematological, and endocrine diseases; (d) Symptomatic Parkinson's syndrome, patients with psychosis, substance abuse, and history of alcohol abuse.

The main symptoms of traditional Chinese medicine: refer to the guiding principles of clinical research of new Chinese medicine; the guiding principle of clinical research of new Chinese medicine for treating senile dementia.

1.2 treatment methods

Ginsenoside saponin capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

RSSW capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

YXN5 capsule group: 125 ~ 250mg, 3 times / d, according to the condition, individualized administration.

The composition of the components in each group of capsules is shown in Example 6.

YXN10 capsule group: 250mg/granule, 1 capsule/time, 3 times a day.

Levodopa tablet group: 200 mg/tablet, 1 tablet/time, 3 times a day.

The composition of the components in each group of tablets is shown in Example 8.

Five groups of patients were given ginseng total saponin capsules, RSSC capsules, YXN5 capsules, YXN10 capsules and levodopa tablets. The treatment of each group began after the treatment, and the medication was continued for 15 days, and the drug was shared for 9 months. Use other cerebral vasodilator drugs, brain cell metabolism drugs, neurological function regulating drugs. The efficacy of each group was evaluated after 1, 3, 6, and 9 months of treatment.

1.3 Observation indicators

1 Safety observation: including general life indicators (blood pressure, heart rate, breathing), blood routine, urine routine, heart, liver and kidney function tests and adverse reactions (excitement, irritability, dry mouth, dry tongue). The general life index is twice a day, and the outpatients tell the method, and they and their families make detailed records; other indicators are checked once before and after treatment.

2 curative observation: the main symptoms before and after treatment (finger and limb flexion, shaking, muscle stiffness caused by a certain position of the limb or all limbs can not be autonomous activities), physical signs, laboratory indicators (hemorheology, cerebral blood flow diagram , platelet adhesion and aggregation rate) improvement.

Efficacy criteria were evaluated according to the Unified Parkinson's Disease Rating Scale (UPDRS).

Efficacy index = (pre-treatment score - post-treatment score) / pre-treatment score x 100%.

The efficacy index ≥85% is cured; 70-84% is significant; 20%-69% is effective; <20% is ineffective.

1 clinical cure: (the finger and limbs flutter, shaking, muscle stiffness caused by a certain position of the limb or all limbs can not be autonomous activities) and other symptoms disappear, you can walk independently, life can take care of themselves, upper limbs and lower limbs muscle strength recovered to 4-5 level.

2 markedly effective: the finger and limbs vibrate, shake, muscle stiffness caused by a certain position of the limb or all limbs can not be autonomous activities and other symptoms are significantly improved, can walk on foot, upper limbs and lower extremity muscle strength recovery level 2 or above.

3 Effective: Fingers and limbs vibrate, shake, muscle stiffness caused by a certain multiple of the limbs or all limbs can not be autonomous activities and other symptoms improved, the upper limbs and lower limbs muscle strength recovered to level 1 or above.

4 Invalid: before and after treatment, the fingers and limbs were vibrated, shaken, muscle stiffness caused by a certain multiple of the limbs or all limbs could not move autonomously and other symptoms did not improve.

The results of the treatment of Parkinson's disease (PD) in each group are shown in Tables 16 and 22.

The severity of motor symptoms was assessed using the PD Unified Rating Scale (UPDRS) Part III and the Hoehn-Yahr Rating Scale. Patients with normal life ability were assessed using the PD Unified Rating Scale (UPDRS) Part II and the Daily Living Ability Questionnaire (ADCS-ADL). The patient's spirit, behavior, and mood were assessed using the PD Unified Rating Scale (UPDRS) Part I.

1.4 Statistical methods: t-test was used for measurement data, chi-square test was used for counting data, and Ridit test was used for grade data.

2. Therapeutic results

The results of the changes in the scales before and after treatment in each group are shown in Table 17-21 and Table 23-27.

Table 16 Comparison of the effects of each group on Parkinson's disease (PD)

Note: Compared with ginseng total saponin, ^## P<0.01.

As can be seen from the above Table 16, the effect of YXN5 capsule containing the biloba lactone synergist on PD disease was significantly improved (P<0.05).

Table 17 Changes in energy, behavior, and mood (UPDRS-I) scores before and after treatment

Note: compared with before treatment in each ^{group, * P <0.05, ** P} <0.01, compared with the total ginsenoside capsule, ^# P <0.05.

Table 18 Changes in daily life activity (UPDRS-II) score scale before and after treatment

Note: compared with before treatment in each ^{group, * P <0.05, ** P} <0.01, compared with the total ginsenoside capsule, ^# P <0.05.

Table 19 Changes in post-treatment motor function test (UPDRS-III) score scale

Note: compared with before treatment in each ^{group, * P <0.05, ** P} <0.01, compared with the total ginsenoside capsule, ^# P <0.05.

Table 20 Changes in post-treatment exercise complications (UPDRS-IV) score scale

Note: compared with before treatment in each ^{group, * P <0.05, ** P} <0.01, compared with the total ginsenoside capsule, ^# P <0.05.

Table 21 Changes in Daily Life Satisfaction (LSIB) Rating Scale before and after treatment

Note: compared with before treatment in each ^{group, * P <0.05, ** P} <0.01, compared with the total ginsenoside capsule, ^# P <0.05.

As can be seen from Table 17-21 above, YXN5 capsules containing biloba lactone synergist can significantly improve the mental, behavioral and emotional (UPDRS-I) scores of PD, daily living activities (UPDRS-II), and motor function tests (UPDRS). -III) score, exercise complication (UPDRS-IV) score, quality of life score scale (LSIB) score, indicating that YXN5 capsule containing biloba lactone synergist can significantly increase the brain of ginseng total saponin and saponin The targeted effect of the department has the effects of relieving muscle tremor and muscle stiffness, and the symptoms such as unfavorable language, insomnia, irritability, irritability, and tongue defects are very obvious, which significantly improves the symptoms of Parkinson's disease.

Table 22 Comparison of the effects of each group on Parkinson's disease (PD)

Note: compared with levodopa, ^# P <0.05.

As can be seen from the above Table 22, the YXN10 capsule containing the biloba lactone synergist significantly improved the efficacy of PD (P<0.05).

Table 23 Changes in energy, behavior, and mood (UPDRS-I) scores before and after treatment

Note: ^* P < 0.05, ^** P < 0.01 compared with before treatment, compared with levodopa, ^# P < 0.05.

Table 24 Changes in daily life activities (UPDRS-II) score scale before and after treatment

Note: ^* P < 0.05, ^** P < 0.01 compared with before treatment, compared with levodopa, ^# P < 0.05.

Table 25 Changes in post-treatment motor function test (UPDRS-III) score scale

Note: ^* P < 0.05, ^** P < 0.01 compared with before treatment, compared with levodopa, ^# P < 0.05.

Table 26 Changes in post-treatment exercise complications (UPDRS-IV) score scale

Note: ^* P < 0.05, ^** P < 0.01 compared with before treatment, compared with levodopa, ^# P < 0.05.

Table 27 Changes in Daily Life Satisfaction (LSIB) Rating Scale before and after treatment

Note: ^* P < 0.05, ^** P < 0.01 compared with before treatment, compared with levodopa, ^# P < 0.05.

As can be seen from Tables 23-27 above, YXN10 capsules containing biloba lactone synergist can significantly improve the mental, behavioral and emotional (UPDRS-I) scores of PD, daily living activities (UPDRS-II), and motor function tests (UPDRS). -III) score, exercise complication (UPDRS-IV) score, and quality of life score scale (LSIB) score, indicating that YXN10 capsules containing biloba lactone synergist can significantly improve the brain-targeted efficacy of levodopa, It relieves muscle tremor and muscle stiffness, and is very effective in improving language, insomnia, and other symptoms, and significantly improves the symptoms of Parkinson's disease.

Test Example 3: Application in clinical treatment of vascular dementia

1. Clinical data and methods

1.1 General Information

110 volunteers with vascular dementia were screened, 60 males and 50 females; aged 58-74 years, mean 63.1 years; the shortest course was 1 year, the longest was 11 years, and the average was 3.6 years. After clinical examination, neurological test, and confirmed by head CT or MR. Volunteer patients were randomly divided into ginseng total saponin capsule group, RSSW capsule group, YXN5 capsule group, YXN7 tablet group and vinpocetine tablet group, with 22 people in each group.

Among them, the diagnostic criteria: the diagnostic criteria for vascular dementia in DSM-IV.

Exclusion criteria: severe neurological, blood, endocrine and other primary diseases and the Haijinsky ischemic index (HIS), total score 18 Points, score <7 is divided into senile dementia.

Scale selection: 1. American simple intelligence scale, total score of 30 points, if the score <16 points for intelligent obstacles; 2. Japan Hasegawa dementia scale, total score of 30 points, if the score <16 points for dementia ; 3. Haijinsky ischemic refers to the number of tables (HIS), a total score of 18 points, if the score is > 7 points for vascular dementia, score <7 points for senile dementia.

Combined with clinical experience, the following symptoms are used as observation indicators: the expression is sluggish, the language is unfavorable, or the language is inferior or the language is wrong, forgetting, not swearing, dizziness, headache, and tongue licking.

Efficacy criteria: The comprehensive assessment method was adopted to take the changes in the intelligence state and physical signs before and after treatment as the comprehensive review content, with the focus on intelligent change. The recovery score of the recovery of the Hasegawa dementia scale increased to a normal value, the score of the effective person increased by 5 points or more, and the score of the effective person increased by less than 5 points, and the score of the invalid person not only increased but decreased.

1.2 treatment methods

Ginsenoside saponin capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

RSSW capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

YXN5 capsule group: 125 ~ 250mg, 3 times / d, according to the condition, individualized administration.

The composition of the components in each group of capsules is shown in Example 6.

YXN7 tablet group: 5 mg/tablet, 1 tablet each morning and evening.

Vinpocetine tablet group: 3.57 mg/tablet, 1 tablet each morning and evening.

The composition of the components in each group of tablets is shown in Example 7.

The above five groups of patients were given ginseng total saponin capsules, RSSW capsules, YXN5 capsules, YXN7 tablets and vinpocetine tablets. The treatment of each group was started after the treatment, and the drug was administered continuously for 2 months as a course of treatment. During the course of treatment, other cerebral vasodilator drugs, brain cell metabolism drugs, and neuromodulation drugs were discontinued.

2. Therapeutic results

Table 28 Changes in the score of the simple smart scale before and after treatment

Group n Before treatment After treatment YXN5 capsule twenty two 13.61±5.13 21.61±5.33 ^*# RSSW capsule twenty two 13.56±5.09 19.15±4.89 ^* Ginseng total saponin capsule twenty two 13.58±5.23 17.91±4.76 ^*

Note: compared with before treatment in each group, ^* P <0.05, with total ginsenoside group, ^# P <0.01.

Table 29 Changes in the scores of the Hasegawa Dementia Scale before and after treatment

Group n Before treatment After treatment YXN5 capsule twenty two 12.83±4.61 19.86±6.13 ^* RSSW capsule twenty two 12.67±4.05 18.92±5.43 ^* Ginseng total saponin capsule twenty two 12.71±4.06 17.89±4.07 ^*

Note: ^* P < 0.05 compared with each group before treatment.

Table 30 Comparison of the effects of each group on vascular dementia

As can be seen from Table 28-30 above, YXN5 capsules, RSSW capsules and ginseng total saponins capsules have significant effects on the intelligence of vascular dementia. Among them, YXN5 capsule containing biloba lactone synergist has the best effect on vascular dementia, indicating that YXN5 capsule containing biloba lactone synergist can significantly improve the brain-targeting effect of drug active molecules and fully exert its vascularity. Therapeutic enhancement of dementia.

Table 31 Changes in the scores of the simple smart scale before and after treatment

Drug group n Before treatment After treatment YXN7 piece twenty two 12.21±5.16 21.45±6.51 ^** Vinpocetine tablets twenty two 12.31±5.24 19.79±6.01 ^*

Note: ^* P < 0.05 compared with each group before treatment.

Table 32 Changes in the scores of the Hasegawa dementia scale before and after treatment

Drug group n Before treatment After treatment YXN7 piece twenty two 12.05±5.35 20.91±5.23 ^** Vinpocetine tablets twenty two 12.01±5.74 19.05±5.43 ^*

Note: ^* P < 0.05 compared with each group before treatment.

Table 33 The efficacy of each group on vascular dementia

As can be seen from the above Tables 31-33, YXN7 tablets and Vinpocetine tablets have a significant improvement in the intelligence of vascular dementia. Among them, YXN7 tablets containing biloba lactone synergist have the best effect on vascular dementia, indicating that YXN7 tablets containing biloba lactone synergist can significantly improve the brain-targeting effect of vinpocetine and fully exert its vascularity. Therapeutic enhancement of dementia.

Test Example 4: Application in clinical treatment of senile dementia

Forty-five volunteer patients with Alzheimer's disease (AD; Alzheimer's disease) with a score of <7 in the Haijinsky Ischemic Index (HIS) were randomly divided into 3 groups, 15 in each group. They were ginseng total saponin capsule group, RSSW capsule group and YXN5 capsule group. The dosing schedule for each group is as follows:

Ginsenoside saponin capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

RSSW capsule group: 250mg/granule, 2 capsules/time, 3 times a day.

YXN5 capsule group: 125 ~ 250mg, 3 times / d, according to the condition, individualized administration.

The composition of the components in each group of capsules is shown in Example 6.

Three groups of patients were given ginseng total saponin capsules, RSSW capsules and YXN5 capsules. The treatment of each group began after the treatment, and the treatment was continued for 2 months as a course of treatment. All the treatments were performed for 3 courses, during which other cerebrovascular dilatation drugs were discontinued. Brain cell metabolism drugs, neurological function regulating drugs, the results are shown in Table 34.

Table 34 Comparison of the effects of each group on senile dementia

As can be seen from Table 34, YXN5 capsules have the best effect on the treatment of senile dementia, with a total effective rate of 73.33%, which is superior to ginseng total saponin capsules, indicating that YXN5 capsules containing biloba lactone synergist can significantly increase the activity of medicinal molecules. The brain is targeted for efficacy and fully exerts a therapeutic effect on senile dementia.

The above is only a preferred embodiment of the present invention, and it should be noted that the above-described preferred embodiments are not to be construed as limiting the scope of the invention, and the scope of the invention should be determined by the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and improvements can be made without departing from the spirit and scope of the invention.

Claims (10)

The use of ginkgolide as a synergist in the preparation of a medicament or health care product for preventing and treating cranial nerve injury diseases. The use according to Claim 1, characterized in that the ginkgolide is at least one of a natural product of a lactone lactone, a ginkgolide extract, a natural product ginkgolide or a hydroxy derivative of a ginkgolide extract. One. The use according to claim 2, characterized in that the hydroxy derivative of the natural product ginkgolide or biloba lactone extract is a glycoside, ester or ether. The invention according to claim 1 or 2, wherein the ginkgolide extract is at least one of a ginkgolide extract or a ginkgo biloba extract, and the ginkgolide extract is ginkgo. The content of lactone is from 1 to 99%. The use according to claim 1, wherein the pharmaceutically active molecule having a therapeutic or health-care effect on a cranial nerve injury disease is selected from the group consisting of ginseng total saponins, Shouwu stilbene glycosides, resveratrol, and left-handedness. At least one of dopa, edaravone, vinpocetine, nigralin, citicoline or oxiracetam. A pharmaceutical or health care product composition for preventing and treating a brain nerve injury disease, characterized in that the pharmaceutical or health care product composition comprises a biloba lactone synergist and a pharmaceutical activity having a therapeutic or health-care effect on a brain nerve injury disease molecule. The pharmaceutical or nutraceutical composition according to claim 6, wherein the ginkgolide synergist is a natural product of ginkgolide, ginkgolide extract, natural product ginkgolide or ginkgolide extract. At least one of the hydroxy derivatives of the substance; The medicinal active molecule having therapeutic or health-care effects on cranial nerve injury diseases is selected from the group consisting of ginseng total saponins, Shouwu stilbene glycosides, resveratrol, levodopa, edaravone, vinpocetine, and nitrite At least one of ergoline, citicoline or oxiracetam; The hydroxy derivative of the natural product ginkgolide or biloba lactone extract is its glycoside, ester or ether. The pharmaceutical or nutraceutical composition according to claim 6 or 7, wherein the pharmaceutical or nutraceutical composition comprises: biloba lactone synergist, ginseng total saponin and shouwustilbene glycoside, The weight ratio of the biloba lactone synergist, the ginseng total saponin and the Shouwu stilbene glycoside is 1:10 to 60:10 to 50; Or a ginkgolide synergist, ginseng total saponin and resveratrol, the weight ratio of the biloba lactone synergist, ginseng total saponin and resveratrol is 1:10 to 25:10-20; Or the biloba lactone synergist, vinpocetine, the weight ratio of the biloba lactone synergist, vinpocetine is 1:10-25; Or the concentration of the biloba lactone synergist, levodopa, the biloba lactone synergist, levodopa is 1:20-50; Or the biloba lactone synergist, ergoline, the weight ratio of the biloba lactone synergist, nigeroline is 1:10-50; Or the biloba lactone synergist, oxiracetam, the weight ratio of the bilobalide synergist, oxiracetam is 1:20-50; Or the biloba lactone synergist, edaravone, the weight ratio of the biloba lactone synergist, edaravone is 1:20-50; Or the biloba lactone synergist, citicoline, the weight ratio of the biloba lactone synergist and citicoline is 1:20-50. The pharmaceutical or nutraceutical composition according to claim 6, wherein the cranial nerve injury disease is a cerebral ischemic injury disease or a neurodegenerative disease. The pharmaceutical or nutraceutical composition according to claim 9, wherein the cerebral ischemic injury disease is ischemic stroke, chronic cerebral ischemia, stroke sequelae or vascular dementia; said neurodegenerative disease For senile dementia, Parkinson's disease, Alzheimer's disease or mild cognitive impairment.

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