TWI783161B - Use of a sunflower extract for preparing compositions for preventing or treating nervous system diseases - Google Patents

Abstract

A use of a sunflower extract for the preparation of a composition for preventing or treating nervous system diseases, wherein the sunflower extract is extracted from plants of the genus Sunflower, and contains polysaccharide components; wherein the nervous system diseases include neurodegenerative diseases and cell cycle Abnormal, where the neurodegenerative disease includes dementia or motor disorder, the dementia includes Alzheimer's disease, vascular dementia, frontotemporal dementia, Lewy body dementia, or mild cognitive Functional disorder, the motor disease includes Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, cerebellar atrophy or multiple system atrophy, wherein the cell cycle abnormality of nervous system cells is a nervous system tumor.

Description

Use of a sunflower extract for preparing compositions for preventing or treating nervous system diseases

The present invention relates to the use of a sunflower extract for preventing or treating nervous system diseases.

The nervous system is composed of a network of specialized cells such as glial cells (including astrocytes, oligodendrocytes, and microglia) and neurons, which transmit signals between different parts of the body and receive external and internal The information is integrated and coordinated to instruct the body to make an appropriate response, so that the organism can carry out rapid and short-term information transmission to protect the individual's survival and continuation of life. However, due to the inability of the vast majority of nerve cells to regenerate, coupled with the high complexity of the neural network system, neurological diseases affect a wide range of people to varying degrees and are often incurable.

Nervous system diseases include neurodegenerative diseases and cell cycle abnormalities of nerve cells. With the aging of the population, the incidence of neurodegenerative diseases is increasing. How to improve and treat neurodegenerative diseases is the current research important topic. Neurodegenerative diseases include dementia and movement disorders, both of which are directly or indirectly related to mitochondrial health.

Dementia is a chronic neurodegenerative disease, in which the patient's thinking ability and memory experience long-term and progressive degeneration, which eventually affects the daily life functions of the patient and even the caregiver. In addition to cognitive function and memory deterioration, other common symptoms include emotional problems, language problems, and reduced mobility. Age is the most important risk factor for dementia, but dementia Mental illness is an active disease of the brain rather than a result of aging alone, so the diagnosis of dementia includes a decline in mental function that is more severe than is typically the case with aging. Dementia is not a single disease, as long as there is progressive cognitive decline and memory loss, it may be diagnosed as dementia.

The most common type of dementia is Alzheimer's disease, which accounts for about 50-70% of all dementia patients. Another common form of dementia is vascular dementia, which accounts for about 25% of cases. Vascular dementia is dementia caused by cerebrovascular disease, and its degeneration speed depends on the number of strokes and the location of the stroke. If the amyloid-beta peptide (Aβ) that plays an important role in the pathogenesis of Alzheimer's disease is secreted by vascular endothelial cells and forms senile plaques that accumulate on the blood vessel wall, it is also a blood vessel. One of the important risk factors for dementia. Dementia with Lewy body, which accounts for about 15% of dementia patients, in addition to cognitive impairment, may appear in the early stages of the disease, such as body stiffness, hand tremors, unsteady walking, and repetitive unexplained movements. Falls, and these symptoms are similar to Parkinson's disease; in fact, the core component protein of Lewy bodies is α-synuclein, whose mutations cause autosomal dominant familial Parkinson's disease. In addition to the above-mentioned types of dementia, there is also frontotemporal lobe dementia. The biggest difference between this type of dementia and Alzheimer's disease is that the initial symptoms of the patient are not memory function decline, but Behavior problems, personality changes, or degeneration of language function, and the mutated gene that causes frontotemporal dementia is also different from the gene that causes Alzheimer's disease. In addition to the diagnosed types of dementia, there are also patients with mild cognitive impairment (mild cognitive impairment) before dementia, which is a transitional stage between normal cognitive function and mild dementia, although a small number of patients will return to normal cognitive function, but most patients will further degenerate and develop develop dementia; therefore, patients with mild cognitive impairment belong to the high-risk group of developing dementia in the future. At present, patients with clinical amnesia-type mild cognitive impairment and Alzheimer's disease biomarkers have been diagnosed as MCI (MCI due to Alzheimer's disease) caused by Alzheimer's disease. It could be very early Alzheimer's disease. Currently, there is no effective drug to treat MCI, and the course of the disease can only be slowed down through non-drug treatments such as controlling risk factors or strengthening cognitive function training.

The drugs currently used to treat Alzheimer's disease cannot protect nerve cells from apoptosis, and can only temporarily alleviate the symptoms of the patient's disease. Therefore, there is still no cure for Alzheimer's disease. Currently prescription drugs for Alzheimer's disease fall into two categories: acetylcholinesterase inhibitors and NMDA receptor antagonists, but both There are many side effects; acetylcholinesterase inhibitors are suitable for patients with mild to moderate disease. These drugs can inhibit the decomposition of acetylcholine, thereby increasing its content in the brain, which can improve the patient's cognition, and its side effects Including urinary incontinence, bradycardia, myopia, etc.; NMDA receptor antagonists, by blocking the excitotoxicity (excitotoxicity) caused by glutamate stimulating NMDA receptors, and then play a therapeutic role, suitable for moderate to severe diseases or not suitable for use Common side effects in Alzheimer's patients treated with acetylcholinesterase inhibitors are dizziness, drowsiness, headache, and constipation. However, there is no cure for this neurodegenerative disease, regardless of its symptom-relieving effects and its side effects.

Generally, cells are in the G0-phase when they are in the state of differentiation, while cells in the state of division and proliferation go through four different cycles (G1-S-G2-M) in order to complete mitosis. Cells in each division cycle will express specific proteins, which can be detected by immunofluorescence staining, for example, G1-phase expresses cyclin-D1, S-phase expresses proliferating cell nuclear antigen (PCNA), G2/M-phase expression phosphor-Histone H3, apoptosis can be represented by cleaved caspase-3. It is known that differentiated nerve cells will stop dividing and maintain in G0-phase under normal living conditions, but there is a lot of evidence that fully differentiated mature neurons will try to re-enter the cell cycle when stimulated or under stress, but Due to the inability to go through mitosis, it changes to apoptosis. Therefore, if neurons can be inhibited from re-entering the cell cycle, they should be able to inhibit cell apoptosis and cause neuroprotection.

Clinical studies have shown that neurons expressing cell cycle-related proteins (such as cyclin-D1) can be detected in the hippocampus and cerebral cortex of patients with Alzheimer's disease after postmortem autopsy, but in the brains of normal controls at the same age. There will be no similar situation, showing that the death of mature and differentiated neurons after entering the cell cycle is a phenomenon that can be observed clinically, and not just an artificial phenomenon that occurs in animals or in cell culture.

Another common neurodegenerative disease is Parkinson's disease, which is a movement disorder and is more likely to occur in people over the age of 50 or 60. The symptoms are resting tremors, muscle stiffness, slow movements, and unsteady posture leading to balance disorders. Mainly. Early symptoms often include regular shaking of one hand, slow movements, and muscle stiffness gradually spread throughout the body. In the final stage, it is almost impossible to move around in a wheelchair. Parkinson's disease is due to the lack of dopamine due to the death of neurons that release dopamine as a neurotransmitter in the substantia nigra pars compacta of the midbrain, which in turn makes the basal ganglia (basal ganglia) dependent on substantia nigra cells to provide Dopamine, as a neurotransmitter, is affected in the striatum, which cannot regulate the messages of the cerebral cortex, thalamus, and extrapyramidal system, thus causing dysfunction in the coordination and operation of various muscles. Like the aforementioned Alzheimer's disease, Parkinson's disease cannot be cured. Currently, L-dopamine supplementation is used clinically to alleviate the symptoms of patients with Parkinson's disease, but the use of L-dopamine It often causes side effects; when the symptoms are more severe and cannot be controlled by drugs, implanting electrodes in the brain for deep brain stimulation (deep brain stimulation) can also be considered, but this is a brain surgery and is highly invasive and uncertain. And because neurons continue to die, no matter whether drugs or surgery can only alleviate some symptoms at most, they have no therapeutic function.

Mitochondrial dysfunction and oxidative stress have been proven to be important causes of dopaminergic neuron death, and thus become an important pathogenic mechanism of Parkinson's disease. In fact, several pathogenic genes (such as PINK1, Parkin, DJ-1, etc.) that lead to familial Parkinson's disease are closely related to maintaining the normal function of mitochondria. PINK1 and Parkin are directly involved in the "quality control" of mitochondria in cells. Mitochondria with damaged structures or poor function due to aging will clear them through the joint action of PINK1 and Parkin, and decompose the components such as amines amino acids, etc. are recycled and reused, while DJ-1 is an antioxidant protein located in mitochondria. In addition to familial Parkinson's disease, previous experiments have confirmed that some patients with sporadic Parkinson's disease have lower leukocyte mitochondrial function than normal people, especially the activity of mitochondrial electron transport chain complex I; in other words , these patients with sporadic Parkinson's disease may have defects in their mitochondrial function throughout the body (this may be due to the polymorphism of their mitochondrial DNA), just because the neurons in the black nucleus compact area use Dopamine is a neurotransmitter, and the synthesis process will generate more free radicals to cause oxidative stress. Therefore, these dopamine neurons accumulated over decades are the first to be damaged, and then Parkinson's symptoms appear. Regardless of its upstream molecular mechanism, mitochondrial dysfunction is a very important pathological mechanism of Parkinson's disease, so inhibitors of mitochondrial electron transport chain complex I (such as MPTP or rotenone) are often used in animal experiments. Or cell experiments are used as a drug model to simulate Parkinson's disease, and drugs or natural extracts that can protect against the neurotoxicity of mitochondrial inhibitors are expected to be helpful to Parkinson's disease patients beneficial.

3-nitropropionic acid (3-NP), an irreversible inhibitor of mitochondrial complex II, mainly affects the effect of succinate dehydrogenase, and is easier to selectively damage the striatum, but the dose is high Eventually damages the cerebral cortex as well. When administered to primates or rodents, the pathological features induced by 3-NP are very similar to the clinical features of Huntington's disease. In addition to Huntington's disease and the aforementioned Alzheimer's disease and Parkinson's disease, mitochondrial dysfunction is also directly related to a variety of other neurodegenerative diseases, such as spinal cord lateral sclerosis and cerebellar atrophy Diseases; in fact, in addition to chronic neurodegenerative diseases, the pathogenesis of acute nervous system injuries such as ischemic stroke and spinal cord injury also includes mitochondrial dysfunction or damage. Therefore, drugs or preparations that can protect nerve cells from damage to mitochondrial function should have the potential to treat or prevent neurological diseases.

Schizophrenia is a kind of psychiatric disease. Its pathological mechanism is complex and still unknown. The common cause of it is the abnormality of dopamine neurons. The availability of striatal dopamine transporters is related to its treatment status.

Fully differentiated nerve cells should be in a quiescent state in a normal healthy state and no longer undergo mitosis; however, during the progression of neurodegenerative diseases, differentiated neurons try to re-enter the cell cycle, but fail to go to the cell cycle The phenomenon of apoptosis has been confirmed from clinical and animal cell experiments. Therefore, the subsequent apoptosis caused by abnormal neuronal cell cycle is one of the important pathogenic mechanisms in neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, and it can inhibit the abnormal cell cycle and apoptosis of neurons. Drugs or natural extracts of these drugs should be expected to help patients with neurodegenerative diseases. On the other hand, the cause of cancer is the abnormal proliferation and metastasis of cancer cells. The current treatment methods include surgical resection, radiation therapy and chemotherapy drugs. Tumors have uncontrolled cell division due to the continuous entry of cancer cells into the cell cycle, which is associated with the inhibition of neuronal Cells entering the cell cycle will inhibit apoptosis and cause neuroprotection. On the contrary, most drugs that inhibit tumor cell division often lead to apoptosis, which is beneficial to cancer treatment. Therefore, if a drug or natural product that inhibits cell division can be found, it may have anti-cancer effects effect. Tumors in the nervous system include head and neck cancer, brain tumor, peripheral nervous system cancer, central nervous system cancer, acoustic neuroma, oligodendroglioma, meningioma, neuroblastoma, etc.

At present, the clinical effect of drugs for neurological diseases is limited and has serious side effects, and many patients cannot continue treatment. More importantly, the medicine only alleviates the symptoms, but fails to fundamentally solve the problem of neurodegeneration and death, so how to develop a new ingredient that can truly prevent or treat neurological diseases is an important issue to be solved by the present invention.

In view of the aforementioned shortcomings in the prior art, the object of the present invention is to provide a use of a sunflower extract for the preparation of a pharmaceutical composition for preventing or treating nervous system diseases, which is extracted from plants of the genus sunflower.

The object of the present invention is to provide a composition containing the extract of the sunflower for the prevention or treatment of nervous system diseases. drug.

For the purpose of the aforementioned invention, wherein the nervous system disease is a neurodegenerative disease, including dementia or motor disorder.

For the purpose of the aforementioned invention, wherein the dementia includes Alzheimer's disease, vascular dementia, frontotemporal dementia, Lewy body dementia or mild cognitive impairment.

To achieve the purpose of the aforementioned invention, wherein the neurological disease is a neurological disease related to mitochondrial damage.

To achieve the purpose of the aforementioned invention, wherein the neurological disease related to mitochondrial damage is a neurological disease related to damage to the substantia nigra or striatum.

To achieve the aforementioned object of the invention, the treatment of Alzheimer's disease includes delaying the onset of cognitive impairment or delaying the onset of memory impairment in Alzheimer's disease.

For the purpose of the aforementioned invention, wherein the motor disease includes Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, cerebellar atrophy or multiple system atrophy.

For the purpose of the aforementioned invention, wherein the nervous system disease is cell cycle abnormality.

For the purpose of the aforementioned invention, wherein the cell cycle abnormality of the nervous system cells is a nervous system tumor.

In order to achieve the purpose of the aforementioned invention, the extraction steps of the fallen sunflower extract are as follows: step 1, fresh or frozen fallen sunflowers are directly ground without adding water, or fresh or frozen fallen sunflowers are added with water or alcohol to grind and stir; step 2, filter to remove residues Obtain the succulent juice; step 3, add alcohol to precipitate, and filter to obtain the precipitate.

In order to achieve the purpose of the aforementioned invention, wherein water is added before stirring in step 1, the weight ratio of the fallen sunflower to water is 1:1~1:20, the stirring temperature is 70°C~121°C, and the stirring time is 10~200 minutes; The alcohols in step 3 are ethanol, glycerin, propylene glycol (Propylene Glycol), propanol, 1,3-butanediol (1,3-Butylene Glycol), isobutanol, isoamyl alcohol, butanol, hexanol , Octyl Alcohol, Lauryl Alcohol (Lauryl Alcohol), 1-Cetyl Alcohol, Benzyl Alcohol, Geraniol, Linalool, Menthol, 3-Phenylpropanol, 1-Phenylpropanol, Nerol, Perillyl Alcohol , the volume ratio of the fallen sunflower juice to the alcohol is 1:1~1:6, and it is allowed to stand and precipitate at a low temperature below 10°C; it can be further concentrated between the step 2 and the step 3; after the step 3, it can be Carry out second alcohol precipitation further, obtain secondary precipitate, alcohol is ethanol, glycerin Oil, Propylene Glycol, Propanol, 1,3-Butylene Glycol, Isobutanol, Propanol, Isoamyl Alcohol, Butanol, Hexanol, Octanol, Ladecanol (Lauryl Alcohol), 1-Cetyl Alcohol, Benzyl Alcohol, Geraniol, Linalool, Menthol, 3-Phenylpropanol, 1-Phenylpropanol, Nerol, Perillyl Alcohol.

In order to achieve the purpose of the aforementioned invention, wherein the sunflower extract contains polysaccharides.

In summary, the present invention obtains the extract from edible plants of the genus Artholium, which can: (1) protect neurotoxicity caused by Aβ or 3-NP, (2) protect neurotoxicity caused by Aβ or 3-NP Dendritic damage, (3) inhibit Aβ-induced neuron re-entry into the cell cycle and subsequent neuronal apoptosis in differentiated neurons, (4) reduce cognitive and memory decline caused by dementia, and achieve Prevention and treatment of neurodegenerative diseases, including stroke, mild cognitive impairment, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, multiple sclerosis, cerebellar atrophy or psychosis Efficacy in dyslexia.

S01~S05‧‧‧Sunflower Extraction Process

Fig. 1 is the extraction process of falling sunflower.

Fig. 2A is the effect of sunflower extract (50mg/kg) on the cognition and memory of Alzheimer's disease mice.

Fig. 2B is the effect of the sunflower extract (25mg/kg) on the cognition and memory of Alzheimer's disease mice.

Figure 3 shows the effect of different batches of sunflower extracts on the neurotoxicity of Aβ.

Figure 4A is the neurotoxicity of Aβ caused by the co-treatment of different concentrations of sunflower extracts influences.

Fig. 4B is the effect of pretreatment with different concentrations of sunflower extracts on the neurotoxicity of Aβ.

Fig. 4C is the result of confocal microscopy of the effect of the sunflower extract on the damage of neuron dendrites caused by Aβ.

Fig. 4D is the analysis of the total length of dendrites of the results of confocal microscopy of the effect of the sunflower extract on the damage of neuron dendrites caused by Aβ.

FIG. 4E is the analysis of the dendrite length of a single nerve cell in the results of confocal microscopy of the effect of the sunflower extract on the damage of neuron dendrites caused by Aβ.

Fig. 4F is the analysis of the number of total dendrite forks of the results of confocal microscopy of the effect of the sunflower extract on the damage of neuron dendrites caused by Aβ.

Fig. 4G is the analysis of the number of dendrites of a single nerve cell by confocal microscopy of the effect of the sunflower extract on the damage of neuron dendrites caused by Aβ.

Fig. 5 is the effect of sunflower extract on the oxidative stress caused by Aβ.

Fig. 6 is the effect of sunflower extract on apoptosis caused by Aβ.

Figure 7A uses immunofluorescence staining method to detect the effect of sunflower extract on the expression of cyclin-D1 in neurons after Aβ stimulates differentiated neurons to re-enter cell cycle G1-phase.

FIG. 7B is a western blotting method to detect the effect of sunflower extract on the expression of G1-phase marker protein cyclin-D1 in cerebral cortex neurons stimulated by Aβ.

Figure 7C uses immunofluorescence staining to detect the effect of sunflower extract on the expression of p-Rb in neurons after Aβ stimulates differentiated neurons to reenter cell cycle G1-phase.

Figure 7D is to detect the effect of sunflower extract on Aβ stimulation of cerebral cortex by Western blot method. The effect of the G1-phase marker protein p-Rb expressed by cells.

Figure 7E uses immunofluorescence staining to detect the effect of sunflower extract on stimulating differentiated neurons to re-enter cell cycle S-phase and expressing PCNA in neurons.

FIG. 7F detects the effect of sunflower extract on the expression of S-phase marker protein PCNA in cerebral cortex neurons stimulated by Aβ by Western blotting method.

Fig. 7G is a western blotting method to detect the effect of sunflower extract on the expression of G2/M-phase marker protein p-Histone H3 in cerebral cortex neurons stimulated by Aβ.

Fig. 7H is to detect the effect of the post-treatment of the sunflower extract on the expression of the G1-phase marker protein cyclin-D1 in the cerebral cortex neurons stimulated by Aβ by Western blot method.

FIG. 7I is a Western blotting method to detect the effect of the post-treatment of the sunflower extract on the expression of the G1-phase marker protein p-Rb in the cerebral cortex neurons stimulated by Aβ.

Fig. 7J is a western blotting method to detect the effect of the post-treatment of the sunflower extract on the expression of the S-phase marker protein PCNA in the cerebral cortex neurons stimulated by Aβ.

FIG. 7K is a western blotting method to detect the effect of post-treatment of sunflower extract on the expression of G2/M-phase marker protein p-Histone H3 in cerebral cortex neurons stimulated by Aβ.

Figure 7L is the western blotting method to detect the effect of the post-treatment of the sunflower extract on the expression of the apoptosis marker protein cleaved caspase-3 in the cerebral cortex neurons stimulated by Aβ.

Fig. 8 is the effect of different batches of the sunflower extract on the neurotoxicity of 3-NP.

Fig. 9A is the result of confocal microscopy of the cell death caused by 3-NP caused by the extract of sunflower.

Fig. 9B is the effect of different concentrations of the extracts of falling sunflowers on the neurotoxicity of 3-NP.

Fig. 9C is the effect of neurotoxicity on 3-NP caused by pretreatment of different concentrations of sunflower extracts influences.

Figure 9D is the result of confocal microscopy of neuron dendritic damage caused by 3-NP extracted from sunflower seedlings.

Fig. 9E is the analysis of the total length of dendrites of the results of confocal microscopy of the effect of sunflower extract on the damage of neuron dendrites caused by 3-NP.

Fig. 9F is the analysis of the number of dendrite total forks in the results of confocal microscopy results of the effect of sunflower extract on the damage of neuron dendrites caused by 3-NP.

Fig. 9G Effects of the sunflower extract and 3-NP on the expression of the post-synaptic protein PSD-95.

Fig. 9H is the effect of the sunflower extract on the oxidative stress caused by 3-NP.

Fig. 9I is the effect of the sunflower extract on the apoptosis caused by 3-NP.

The novel technical features of the present invention, including specific features, are disclosed in the scope of the patent application. For the technical features of the present invention, a better understanding is hereby combined with the description, embodiments based on the principles of the present invention, and drawings to describe the preferred embodiments of the present invention Detailed description.

Unless otherwise defined, all technical and scientific terms mentioned in the description of the invention and the scope of the patent application shall be defined according to the following description. The singular terms "a", "an", and "the" may refer to more than one object, unless otherwise specified. Unless otherwise stated, all technical and scientific terms used in this specification, unless otherwise defined, have the meanings commonly understood by those skilled in the art. "Or", "and", "and" used in this manual, unless otherwise stated, all refer to "or/and". In addition, the terms "comprising" and "including" are not limited open conjunctions. The preceding paragraphs are systemic references only and should not be construed as Limitation on Invention Subject Matter. Unless otherwise stated, the materials used in the present invention are commercially available and readily available.

The term "effective amount" or "therapeutically effective amount" used in this specification refers to a sufficient amount of one of the compounds or drugs that can alleviate one or more disease symptoms or physiological conditions after taking the drug; the result Intentional changes to reduce and/or alleviate signs, symptoms, or causes, or other physiological systems. For example, a therapeutically "effective amount" includes a dose of a compound provided by the present invention that can clinically significantly reduce the symptoms of a disease. An appropriate effective amount, the effective value of which depends on common pharmaceutical techniques, such as dose escalation methods.

The term "composition" used in this specification refers to a product that is derived from mixing or combining more than one active ingredient, and the active ingredient of the product is a combination drug or a composition that is not a combination drug. The term "combination drug" indicates that the active ingredient and its co-agent (co-agent) are administered simultaneously in one dose; the term "non-combination drug" indicates that the active ingredient and its co-agent are separated. The medicaments are administered simultaneously, separately, or sequentially to the subject without limiting the intervals of administration, the aforementioned administration provides the subject with an effective amount of multiple compounds in the body; wherein the non-combination drug can be applied to cocktail therapy, that is, administering three or Various active ingredients.

The terms "treating", "treating" and "therapy" in this specification include alleviating, alleviating, or improving at least one disease symptom or physiological condition, preventing additional symptoms, and inhibiting the disease or physiological condition by means of treatment or prevention , preventing or slowing down the progression of a disease, causing reversion to a disease or condition, slowing down a condition caused by a disease, or stopping the symptoms of a disease or condition.

The term "about" in this specification refers to ±20% of a specified value, or preferably ±10%.

The term "alcohol" used in this manual refers to ethanol, glycerin, propylene glycol (Propylene Glycol), propanol, 1,3-butanediol (1,3-Butylene Glycol), isobutanol, propanol, isoamyl alcohol, butanol, hexanol, octanol, lauryl alcohol ), 1-Cetyl Alcohol, Benzyl Alcohol, Geraniol, Linalool, Menthol, 3-Phenylpropanol, 1-Phenylpropanol, Nerol, Perillyl Alcohol.

Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, gastrointestinal, ocular, pulmonary, mucosal penetration, skin penetration, vaginal, ear, nasal, topical administration.

The following treatment implementations are only examples, and due to the great variability of individual treatment courses, it is not unusual for a large number of deviations from the recommended values. Dosages may vary according to variations not limited to the activity of the compound employed, the disease or physiological state being treated, mode of administration, individual requirements, severity of the disease, and the judgment of the physician.

The toxicity and curative effect of the course of treatment can be determined by standard medical procedures of cell culture or animal experiments, including but not limited to judging the half effective dose (ED ₅₀ , the dose at which the therapeutic effect reaches half of the test subjects and has curative effect).

The raw material used in the present invention is Luokui ( Basella alba L.), also known as imperial palace dish, fungus dish, Chan dish, bean curd dish (Yunnan) and the like. The extraction method of fallen sunflowers is shown in Figure 1 and Table 1: Step 1, ground and stirred fallen sunflowers (S01): fresh or frozen fallen sunflowers are ground and stirred with or without water, and the weight ratio of fallen sunflowers to water is 1:1~ 1:20, the weight of the frozen sunflower is calculated based on the weight before freezing; step 2, high-temperature extraction (S02): heating at 70°C-121°C for 10-200 minutes; step 3, filter to remove the residue to obtain the juice of the sunflower (S03); Step 4. The first ethanol precipitation to obtain the precipitate (S04): add ethanol, the volume ratio of the fallen sunflower juice to ethanol is 1:1~1:6, let stand at a low temperature below 10°C, and filter to obtain the precipitate; step 5 , Freeze-drying to obtain the extract powder (S05) of the sunflower extract.

The aforementioned step 3 can be concentrated after filtering and removing the residue to obtain the fallen sunflower juice; the aforementioned step 4, the first After the first ethanol precipitation obtains the precipitate, the second ethanol precipitation can be carried out. The volume ratio of the precipitate to ethanol is 1:0.5~1:6, and the secondary precipitate is obtained by filtration.

The above-mentioned falling sunflower extract powder contains falling sunflower polysaccharides, as shown in Table 2; the total polysaccharide content is more than 80%, wherein No. 6 is 91.2%, No. 8 is 93.7%, and No. 12 is 91%; the sugar in the total polysaccharide content The uronic acid ratio (uronic acid/total sugar content) varies greatly, among which No. 6 is 48.0%, No. 8 is 52.3%, and No. 12 is 2.8%. (Fructose), mannose (Mannose), rhamnose (Rhamnose), trehalose (Fucose), galactose (Galactose), arabinose (Arabonose), xylose (Xylose), as shown in Table 3, wherein glucose , galactose, Arabinose is the most abundant.

The preparation of the aqueous solution (referred to as PBA) of the falling sunflower extract: the powder of the falling sunflower extract is dissolved in secondary water, and after being dissolved under high temperature and high pressure, the insoluble part is removed by centrifugation. The concentration is 5 mg per milliliter of PBA Sunflower extract powder (contains insoluble fraction).

Example 1: Effect of PBA on Pattern Recognition Novel Object Test Memory (novel object recognition test, NOR) in Alzheimer's disease animals

The animal in vivo test process is shown in Figure 1. Male APPswePS1dE9 Alzheimer's disease model mice were used, and Alzheimer's gene transgenic mice (AD type) and littermate wild-type (wild-type) mice were selected by genetic testing. littermate, ie WT type), were divided into groups at the age of seven months, and centrifuged PBA was given orally (oral gavage), and given three times a week. The NOR test was carried out after one month of continuous administration of the drug (that is, at the age of eight months). During each administration, the changes in individual body weights of the mice were monitored to evaluate the effects of the drug on the physiological functions of the mice.

In this experiment, the mice were divided into four groups, 1. WT: WT mouse control group, not fed PBA (N=16); 2. AD: AD mouse control group, not fed PBA (N=9); 3. . AD+PBA (50mg/kg): AD mouse experimental group, administered 50mg/kg PBA (No. 6, N=9); 4. AD+PBA (25mg/kg): AD mouse experimental group, administered 25mg/kg PBA (No. 6, N=3).

NOR is an animal memory function test experiment that has been used for more than a century. It uses animals (mice in this case) to test the innate desire to explore objects that have never been seen based on vigilance or curiosity. memory capacity of animals. There are many different variations and operating methods of NOR. The core concept is to let the animal explore the same two objects first, and after a period of time, replace one of the objects with a foreign object that has never been seen before. If the memory function of the animal is better, it should be able to remember. One of the objects has been explored before, so it will take a long time to explore the unseen foreign objects. Conversely, if the animal's memory is deficient and does not remember that one of the two objects has been explored, the two different objects are both new and unfamiliar to it, so the proportion of time spent exploring the two objects will be closer. A typical NOR is a memory function evaluation of placing different objects in the same place, and does not involve the concept of space. Therefore, the test results are consistent with working memory, temporal order memory, and episodic-like memory. and other correlations, while less spatial memory (spatial memory) is involved, and can be adjusted according to the length of the operation interval, and the animal's short-term, medium-term and long-term memory can be evaluated at the same time.

The way of operating the NOR test in the embodiment of this case is to place the mice in an open field for ten minutes to explore freely, once a day for three days to make them accustomed to the experimental field, and put them into two identical boxes on the fourth day. The purple sphere (hereinafter referred to as the same object) is allowed to explore for ten minutes. On the fifth day, one of the spheres is replaced with a pink rectangular cylinder (hereinafter referred to as the foreign object) and also given ten minutes of exploration time. On the fourth and fifth days, the whole process of exploration was videotaped, and the video data was manually analyzed to calculate the time for mice to explore individual objects (only frontal sight contact, forelimb contact, nose kiss and whisker contact, The behaviors of climbing objects and biting objects are regarded as exploration actions) The recording unit is the number of seconds. On the fourth day, the data of individuals whose contact time difference between two identical objects is greater than 10 seconds is not included in the calculation. On the fifth day, the total contact time of two different objects must exceed 15 seconds before the data is adopted. The final number of experimental animals adopted (N value ) as above.

The results in Figure 2A show that the value is the absolute value of the fifth day’s foreign object contact time minus the same object contact time divided by the total contact time of the two objects (foreign object contact time plus same object contact time), and the values are all positive numbers. The average score of WT mice is 0.6, which has great curiosity about foreign bodies and can distinguish the same objects. It has been explored yesterday, and taking PBA did not further enhance the scores of WT mice (data not shown); AD The score of AD-type mice was only about 0.4 without taking the drug, compared with the WT-type mice without PBA group, the P value was less than 0.05, which was a statistical difference; while the AD-type mice were fed with 50mg/kg of No. 6 PBA for one After 1 month, its object memory cognitive function has improvement compared with unmedicated AD type group (P value is less than 0.05), and score is close to WT group; Take lower dosage 25mg/kg and AD type mice are fed PBA one month later , its object memory cognitive function seems to be slightly improved compared with the non-medicated AD group, but it may be due to the insufficient number of experimental animals (only 3), so there is a trend but no statistical difference; High dose 100mg/kg No. 6 PBA's memory improvement of AD mice was worse than that of 50mg/kg No. 6 PBA group, and still higher than that of unmedicated AD mice, but statistically there was no significant difference (data not shown ). The body weight monitoring trends of the five groups of mice during the medication period were all the same as those of the WT non-medicated group, and PBA did not significantly affect the weight change of the mice. Based on the above results, PBA has the effect of improving the memory of male APPswePS1dE9 strain Alzheimer's model mice.

The dose used in the aforementioned mouse experiments was 10-100 mg/kg, according to the 2005 US Food and Drug Administration According to the initial estimation method (Estimating the maximum safe starting dose in initial clinical trials for therapeutics in adult healthy volunteers) announced by the Food and Drug Administration, the conversion factor between human and mouse is 12.3 times, and the human dose is 0.8~8.0mg /kg; those with ordinary knowledge in the field of the present invention can obviously know that the foregoing examples are only examples, and the actual effective amount refers to a sufficient amount of the compound or drug, which can alleviate one or more disease symptoms or physiological conditions after taking the medicine. condition; the result is reduction and/or alleviation of signs, symptoms, or causes, or intentional alteration of other physiological systems, the effective value of which depends on usual pharmaceutical techniques, such as drug augmentation methods. Those with ordinary knowledge in the technical field of the present invention can implement it through many transformations and replacements without any difference from the technical features of the present invention. According to the embodiments of the description, the present invention can have various transformations without hindering the implementation. The claims provided in this specification define the scope of the invention, which covers the aforementioned methods and structures and inventions equivalent thereto.

Example 2: The effect of PBA on the neurotoxicity of Aβ in the cellular model The length of Aβ in the human brain varies from about 39 to 43 amino acids, but many studies in the past have proved that artificially synthesized Aβ has 25 to 35 amino acids One amino acid peptide (Aβ25-35) has neurotoxicity; the present invention uses the primary cultured rat cerebral cortex nerve cells as a model, and Aβ25-35 (10μM) is added to PBA (200μg/ml) for 48 hours after treatment. Propidium iodide (PI) staining was used to determine cell viability. The results in Figure 3 show that the single treatment of No. 6 PBA or No. 12 PBA does not affect cell survival, and the survival rate of cells after treatment with Aβ25-35 is less than 50%. No matter whether No. 6 PBA or No. 12 PBA and Aβ25-35 are treated simultaneously, Aβ can be inhibited The resulting cytotoxicity restores the survival of nerve cells, and No. 6 PBA almost completely restores the cell survival rate, achieving the effect of completely protecting nerve cells from Aβ toxicity.

Using the primary cultured rat cerebral cortex neurons as a model, Aβ25-35 After treatment for 48 hours, the reducing ability of mitochondria was measured as an indicator of cell survival. The results in Figure 4A show that the cells were treated with PBA No. 6 and Aβ25-35 for 48 hours at the same time, and the cell survival rate was observed. PBA No. 6 can increase the survival rate in the dose range of 10-300 μg/mL, indicating that it can effectively inhibit the neurotoxicity caused by Aβ25-35 , the effect of inhibiting Aβ25-35 neurotoxicity is dose-dependent, and No. 6 PBA can achieve almost complete protective effect in the dose range of 200-500 μg/mL. Pre-treat with PBA for 8 hours, then add Aβ25-35 for 48 hours, and observe the cell viability. The results in Fig. 4B show that pretreatment with PBA No. 6, PBA No. 7, and PBA No. 8 in the dose range of 300-500 μg/mL can partially inhibit the neurotoxicity caused by Aβ25-35, but it has not achieved complete protection and has no statistical difference.

Co-treated with PBA (200 μg/ml) and Aβ25-35 (10 μM) for 48 hours, and confocal microscopy experiments confirmed that No. 6 PBA restored neuron dendritic damage caused by Aβ25-35 (Figure 4C), and quantified the results Then it was shown that PBA can restore the reduction of the total length of dendrites (Fig. 4D) and the number of forks (Fig. 4F) caused by Aβ25-35, and calculate the length of dendrites (Fig. 4E) and the number of forks (Fig. 4G) also get similar results.

In order to test whether PBA has the effect of anti-oxidation, this experiment uses a reagent CellROX®, which will emit green fluorescence when the cells produce too many free radicals. 200μg/ml) and Aβ25-35 (10μM) were co-treated for 16 hours to find that No. 6 PBA effectively reduced the green fluorescence produced by Aβ25-35, which means that PBA can reduce the production of free radicals and has an antioxidant effect.

It is known that Aβ25-35 can cause cell apoptosis. During the process of cell apoptosis, pro-caspase-3 will be cleaved to produce active cleaved caspase-3. Therefore, the amount of caspase-3 cleavage can represent the indicator of cell apoptosis , In this example, caspase-3 was quantified by western blot method. Figure 6 shows that co-treatment with PBA (200 μg/ml) and Aβ25-35 (10 μM) for 24 or After 48 hours, PBA No. 6 can completely inhibit the caspase-3 cleavage caused by Aβ25-35, which means that PBA has anti-apoptotic effect.

Compared with the cells in the dividing state, the fully differentiated neurons are in the G0 state, but when the nerve cells are stimulated by external pressure, they often try to re-enter the cell cycle, but they cannot go through mitosis smoothly and instead move towards the cell cycle. apoptosis. Using the first generation of cultured cerebral cortical neurons (about 85% of which are neurons, 15% are other cells, such as astrocytes) as a model, this experiment tested whether PBA, an extract of polysaccharides from asparagus, can inhibit amyloid beta-protein (Aβ)-induced re-entry of neurons into the cell cycle and subsequent apoptosis. Immunofluorescence staining was performed after treatment with Aβ25-35 (10 μM) and PBA (250 μg/ml) for 24 hours. In Figure 7A, red represents the nerve cell marker protein MAP-2, green represents the G1-phase marker protein cyclin-D1, and blue The color represents the staining of all nuclei under the microscope by Hoechst staining. It can be observed from the 63 times microscope magnification that Aβ25-35 significantly increases the number of mature neurons that express MAP-2 and are stained with cyclin-D1. , and the 126-fold microscope magnification can be more clearly observed that these cells stained with green cyclin-D1 and red MAP-2 neurons are the same cell, and the quantitative results confirm that PBA can completely inhibit the cyclin-D1 induced by Aβ25-35 ⁺ /MAP-2 ⁺ cell number; In addition to immunofluorescence staining, the results of western blotting in Figure 7B also proved that in the primary cultured cerebral cortical neurons, PBA can completely inhibit the increase of amyloid beta protein cyclin-D1 protein expression. These results show that PBA can completely inhibit the G1-phase of neurons re-entering the cell cycle caused by amyloid beta-protein.

Using p-Rb as another G1-phase marker protein, the experimental results in Figure 7C and Figure 7D show that PBA can completely inhibit the number of p-Rb ⁺ /MAP-2 ⁺ cells caused by Aβ25-35 and the number of cells in the primary cultured brain PBA in cortical nerve cells can completely inhibit the expression of G1-phase marker protein p-Rb protein increased by amyloid beta-type protein.

Taking PCNA as a S-phase marker protein, the experimental results in Figure 7E and 7F show that PBA can completely inhibit the number of PCNA + /MAP-2 + cells caused by Aβ25-35 and that PBA can inhibit the number of PCNA ⁺ /MAP-2 ⁺ cells in the primary cultured cerebral cortex neurons. Completely inhibited the expression of S-phase marker protein PCNA protein increased by amyloid beta-type protein.

The results of western blotting experiments in Figure 7G show that PBA can completely inhibit the expression of the G2/M-phase marker protein p-Histone H3 protein increased by amyloid beta-type protein in the primary cultured cerebral cortical neurons.

When the primary cultured cerebral cortex neurons were treated with amyloid beta-protein Aβ25-35 and asparagus polysaccharide extract PBA, it cannot be excluded that the amyloid beta-protein Aβ25-35 extract and amyloid beta-protein Aβ25-35 could be directly produced in the cell culture medium. The possibility of interaction to inhibit its toxicity; if this is the case, it means that the polysaccharide extract of sunflower has no substantial impact on nerve cells (such as activation of the signal transmission pathway of the neuroprotective mechanism, etc.), so the clinical significance is not great. In order to rule out this possibility, neurons were first treated with Aβ25-35 (10 μM) for 2 hours, then the medium containing Aβ25-35 was removed, and then fresh medium containing PBA (250 μg/ml) was added and treated for another 22 hours , so that Aβ25-35 and PBA will not exist in the medium at the same time, and the two cannot interact. Figure 7H shows that PBA post-treatment for 22 hours in the primary cultured cerebral cortical neurons can completely inhibit the expression of G1-phase marker protein cyclin-D1 protein increased by Aβ25-35 pre-treatment for 2 hours.

Figure 7I shows that PBA post-treatment for 22 hours in primary cultured cerebral cortical neurons can completely inhibit the expression of G1-phase marker protein p-Rb protein increased by Aβ25-35 pre-treatment for 2 hours.

Figure 7J shows that PBA post-treatment for 22 hours in primary cultured cerebral cortical neurons can completely inhibit the protein expression of S-phase marker protein PCNA increased by Aβ25-35 pre-treatment for 2 hours.

Figure 7K shows that post-treatment with PBA for 22 hours in the primary cultured cerebral cortical neurons can completely inhibit the protein expression of the G2/M-phase marker protein p-Histone H3 increased by the pre-treatment of Aβ25-35 for 2 hours.

Figure 7L shows that PBA post-treatment for 22 hours in primary cultured cerebral cortical neurons can completely inhibit the expression of apoptosis marker protein cleaved caspase-3 that was increased by Aβ25-35 pre-treatment for 2 hours.

According to the above results, the results of the cerebral cortex nerve cell model prove that PBA can effectively inhibit the neurotoxicity of amyloid beta protein, promote dendrite regeneration, and has anti-oxidation and anti-apoptosis effects. In addition, no matter through the co-treatment experiment of amyloid-beta-protein and the extract of the polysaccharides of falling sunflower, or the experiment of separate treatment before and after treatment of the extract of amyloid-beta-protein for 2 hours and then of the extract of falling sunflower polysaccharide for 22 hours, the The results have consistently proved that PBA can effectively inhibit the re-entry of differentiated neurons into the cell cycle induced by amyloid beta-protein, thus achieving neuroprotective effects.

Example 3: Effect of PBA on neurotoxicity of 3-NP in cell model

Trinitropropionic acid (3-nitropropionic acid, 3-NP), belongs to the irreversible inhibitor of mitochondrial complex II, mainly affects the effect of succinate dehydrogenase, has a specific effect on striatum, and will cause striatum Injury, the pathological features caused by it are very similar to the clinical features of Huntington's disease; in addition, the pathogenesis of most other neurodegenerative diseases such as Parkinson's disease is also directly related to mitochondrial imbalance or damage. Using primary cultured cerebral cortical neurons as The model was treated with PBA (500 μg/ml) and 3-NP (2.5 mM) for 24 hours and then stained with propidium iodide (PI) to detect cell membrane rupture and stained dead cells to quantify the cell survival rate; The results in Figure 8 show that treating No. 6 PBA or No. 12 PBA alone does not affect the cell survival rate, and the survival rate of the cells after 3-NP treatment is less than 70%. No matter whether it is No. 6 PBA or No. 12 PBA and 3-NP, it can be greatly improved. It significantly reduces the cell death caused by 3-NP and restores the survival of cells. Among them, No. 12 PBA almost completely inhibits the neuroprotective effect of 3-NP toxicity.

To further observe the effects of different concentrations of No. 6 PBA on the neurotoxicity of 3-NP, the first generation of cultured cerebral cortex neurons was also used as a model, and 3-NP (2.5mM) was added to PBA (100-500μg/ml) for 24 hours and then treated with Hoechst staining was used to stain all the cells under the microscope field, and propidium iodide (PI) was used to stain the dead cells under the same field of view, so as to calculate the degree of cell death. In the same field of view, cells and shapes stained by Hoechst can also be simply analyzed to determine cell survival. If the nucleus is evenly stained by Hoechst and becomes oval or round, it is classified as a normal healthy cell. Cells with shrunken nuclei and too bright fluorescent stained by Hoechst were classified as apoptotic or dead cells, and only normal healthy cells were counted. Figure 9A is a representative result of double staining with propidium iodide (PI) and Hoechst (Hoechst), showing that after the primary cultured rat cerebral cortex neurons were treated with 3-NP (2.5mM) for 24 hours, 3- NP significantly increased the number of dead cells stained with PI, and the arrows indicated the nuclei stained with PI, indicating dead cells, while co-treatment with PBA (500 μg/ml) could effectively reduce the degree of cell death caused by 3-NP. Figure 9B is to test the dose-dependence of the protective effect of PBA in the same way. The survival rate is calculated by the number of healthy cells infected by Hoechst, and the control group cells without any treatment is 100%. The results in Figure 9B show that the survival rate remains about half after 3-NP treatment, while the treatment of PBA at the same time can restore cell survival and cause neuroprotection, and it is dose-dependent. The survival rate of fully recovered cells was comparable to that of the control group not treated with 3-NP. In addition to co-treatment, pretreatment with PBA can also cause a protective effect on 3-NP neurotoxicity, and the results in Figure 9C show that pretreatment with PBA (500μg/ml) for 8 hours and then treatment with 3-NP (2.5mM) for 24 hours can partially Both restore cell viability, but the effect is obviously not good together.

Since the primary culture of cerebral cortex cells is a mixed culture system, neurons (neurons) account for about 85% and astrocytes (astrocytes) account for about 15%. Stellate cells and nuclei of all cells were stained for comparison. Cerebral cortex cells were co-treated with 3-NP (2.5mM) and PBA (500μg/ml) for 24 hours (Fig. 9D, Fig. 9E, Fig. 9F), Fig. 9D represents simultaneous staining of MAP-2, GFAP, Hoechst (Hoechst ), MAP-2 exhibits red fluorescence as a marker protein of neurons, GFAP exhibits green fluorescence as a marker protein of astrocytes, Hoechst stains the nuclei of all cells and Shows blue fluorescence. The results in Figure 9D show that neurons are damaged by 3-NP, which can be judged from the atrophy and fracture of dendrites, while PBA can obviously restore the atrophy and fracture of dendrites caused by 3-NP, almost completely restoring its length and complexity . Using software to quantify the total length of dendrites in the same field of view also confirmed that PBA can also increase the total length of dendrites reduced by 3-NP (Fig. dendritic complexity). PSD-95 is a protein on the post-synapse, so its expression can indirectly represent the number of synapses. Nerve cells were treated with 3-NP (2.5mM) and PBA (500μg/ml) for 8 hours at the same time. From the results in Figure 9G, it can be found that 3-NP treatment significantly reduced the expression of PSD-95, but the expression of PSD-95 could be restored by simultaneous treatment of PBA , showing that the decreased number of synapses in neurons affected by 3-NP could be restored by PBA treatment.

PBA has the ability to protect neurons from the toxicity of mitochondrial inhibitors, but its mechanism remains to be explored. The primary cultured cerebral cortical nerve cells were co-treated with 3-NP (2.5mM) and PBA (500μg/ml) for 2 hours, and then CellROX ^® Green reagent was added to detect the reactive oxygen species (reactive oxygen species, ROS, indicated by the arrows) in the cells green fluorescent cells), the results showed that the simultaneous treatment of cells with PBA could significantly reduce the increased ROS generation due to 3-NP (Figure 9H); the primary cultured cerebral cortical neurons were added with 3-NP (2.5mM) and PBA (500μg/ml) Treated for 12 hours, observe the pro-caspase-3, cleaved caspase-3 and tubulin of the cells by western blot method, the results are shown in Figure 9I, caspase-3 is a marker of cell apoptosis, and tubulin is used as an internal control, and the cells can be treated with PBA at the same time. Significantly reduce the cleaved caspase-3 increased by 3-NP, that is to say, PBA can almost completely inhibit the apoptosis of nerve cells caused by 3-NP. These results show that PBA has anti-oxidative and anti-apoptotic effects.

According to the above results, the results of the cerebral cortex nerve cell model prove that PBA can effectively inhibit the neurotoxicity of 3-NP, promote the regeneration of neurite, and have the functions of anti-oxidation and anti-apoptosis and restore the number of synapses.

In the content disclosed in the preferred embodiments of this specification, those with ordinary knowledge in the field of the present invention can clearly understand that the above-mentioned embodiments are only examples; those with ordinary knowledge in the technical field of the present invention can implement them through many transformations and substitutions, and It is not different from the technical features of the present invention. According to the embodiments of the description, the present invention can have various transformations without hindering the implementation. The claims provided in this specification define the scope of the invention, which covers the aforementioned methods and structures and inventions equivalent thereto.

Claims (10)

A use of a sunflower extract for the preparation of a pharmaceutical composition for preventing or treating neurological diseases caused by neurotoxicity or related to mitochondrial damage, wherein the extraction step of the sunflower extract comprises: step 1, heating water extraction Grinding and stirring an asparagus plant; step 2, filtering and removing the residue to obtain an asparagus juice; step 3, after adding ethanol to the asparagus juice for precipitation, filtering to obtain a precipitate, which is the asparagus extract . A food composition, which contains the extract of the sunflower, wherein the extraction step of the extract of the sunflower comprises: step 1, heating water, extracting, grinding and stirring a plant of the sunflower genus; step 2, filtering and removing the residue to obtain the juice of the sunflower ; Step 3, after adding ethanol to the falling sunflower juice, filter to obtain a precipitate, which is the falling sunflower extract; wherein the falling sunflower extract has the function of inhibiting neurotoxicity and mitochondrial damage; wherein The composition comprises health food or animal food. The use as described in Item 1 of the scope of the patent application, wherein the nervous system disease is dementia or motor disorder. The use described in item 3 of the patent application, wherein the dementia is Alzheimer's disease, vascular dementia, frontotemporal dementia, Lewy body dementia or mild cognitive impairment . The use as described in item 1 of the scope of the patent application, wherein the nervous system related to mitochondrial damage The disease is a neurological disorder associated with damage to the substantia nigra or striatum. The use as described in item 1 of the scope of the patent application, wherein the prevention or treatment of neurological diseases caused by neurotoxicity or related to mitochondrial damage is to delay the onset of cognitive impairment or memory impairment in Alzheimer's disease. The use described in item 3 of the patent application, wherein the motor disease is Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, cerebellar atrophy or multiple system atrophy. The use as described in item 1 of the scope of the patent application, wherein the neurological disease is cell cycle abnormality. The use as described in item 1 of the scope of the patent application, wherein the sunflower extract contains polysaccharides. The composition as described in item 2 of the patent application, wherein the sunflower extract contains polysaccharides.

Images