Cistanche peptide composition and application thereof

Abstract

The invention discloses a cistanche peptide composition, which is prepared from the following components in percentage by mass: 15 to 20 percent of cistanche peptide, 10 to 20 percent of brilliant bugleweed herb extract and 8 to 15 percent of radix astragali seu hedysari extract. The invention also provides a process for preparing the cistanche peptide and the cistanche peptide composition, and a traditional Chinese medicine composition containing the cistanche peptide composition. The cistanche peptide composition can be used for preventing or treating neurodegenerative diseases.

Description

Technical Field

[0001] The present invention relates to a Cistanche peptide composition and a preparation process of the Cistanche peptide, and also relates to a traditional Chinese medicine composition comprising the Cistanche peptide composition, and use of the Cistanche peptide composition for preventing or treating neurodegenerative diseases. Background Art

[0002] Neurodegenerative diseases are a class of diseases characterized by progressive degeneration and death of neurons. These diseases usually lead to the gradual loss of cognitive, motor or sensory functions and worsen over time. Common neurodegenerative diseases include Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS). Among them, the number of patients with Parkinson's disease has been increasing year by year worldwide. The disease is characterized by significant degeneration and loss of substantia nigra dopaminergic neurons, gliosis, neuronal α-synuclein (α-Syn) deposition and the appearance of Lewy bodies (LB), which cause patients to experience symptoms such as resting tremor, bradykinesia, muscle rigidity and postural balance disorders. At present, the treatment of Parkinson's disease mainly focuses on drug therapy, such as the use of levodopa and dopamine receptor agonists, which can temporarily relieve symptoms, but long-term use will bring side effects and drug resistance problems.

[0003] Cistanche deserticola, also known as golden shoots, goblin or inch-bright, belongs to the genus Cistanche of the family Orobanchaceae. It is a treasure among traditional Chinese medicinal materials in my country and enjoys the reputation of "desert ginseng". So far, scientists have isolated and identified more than 100 chemical components and trace elements from Cistanche deserticola, including phenylethanol glycosides, proteins, monosaccharides, disaccharides, polysaccharides, amino acids, iridoid glycosides, sterols and polyols. A large number of clinical and basic studies have confirmed that the active ingredients in Cistanche deserticola have significant therapeutic potential for neurodegenerative diseases, especially for Parkinson's disease.

[0004] However, the Cistanche extracts commonly used in the prior art, such as total glycosides of Cistanche, contain a variety of complex chemical components, and the absorption effect of the human body is poor. In addition, due to the complex composition, the interaction between these components may make its mechanism of action difficult to grasp or control, and thus it cannot be used in a targeted manner in the treatment of specific diseases or health improvement.

[0005] Therefore, there is a need in the art to provide a novel composition comprising Cistanche deserticola, which can improve the absorption of the active ingredients of Cistanche deserticola by the human body, and then be specifically used for the treatment of neurodegenerative diseases or health improvement.

[0006] Another advantage of the present invention lies in providing a cistanche peptide composition for use in preventing or treating neurodegenerative diseases, particularly Parkinson's disease.

[0007] In addition, another advantage of the present invention lies in providing a preparation process for cistanche peptides, which can efficiently extract the active ingredients from cistanche. Brief Description of the Invention

[0009] In one aspect, the present invention provides a cistanche peptide composition, which, by mass percentage, comprises the following components: 15%-20% cistanche peptides, 10%-20% extracts of Pterocarpus marsupium, and 8%-15% extracts of Astragalus membranaceus.

[0010] In another aspect, the present invention provides a traditional Chinese medicine composition comprising an effective amount of the aforementioned cistanche peptide composition.

[0011] In another aspect, the present invention provides the use of the aforementioned cistanche peptide composition for preparing a medicament for preventing or treating neurodegenerative diseases. In another aspect, the present invention provides the use of the aforementioned cistanche peptide composition for preventing or treating neurodegenerative diseases. In another aspect, the present invention provides a method for preventing or treating neurodegenerative diseases, which comprises using the aforementioned cistanche peptide composition. In another aspect, the present invention provides a process for preparing the aforementioned cistanche peptides, which comprises the following steps:

[0012] a) Pretreatment: Select fresh cistanche, after washing, slicing, drying, crush it into granules, and then sieve it through an 80-100 mesh sieve to obtain cistanche coarse powder;

[0013] b) Enzymatic hydrolysis: Add the aforementioned cistanche coarse powder to 5-8 times the weight of water, heat it to 80-90°C and stir evenly, after stirring for 10-20 min, cool it down to 40-50°C, add a complex enzyme according to 0.5-2% of the substrate concentration, stir and enzymatically hydrolyze for 2-3 h, after the enzymatic hydrolysis is completed, raise the temperature to 90-100°C and inactivate for 10-20 min, after inactivation and cooling, filter it through a 150-200 mesh filter bag, discharge the filter residue to obtain an enzymatic hydrolysate, and the complex enzyme comprises at least two enzymes selected from the group consisting of cellulase, pectinase, papain, alkaline protease, neutral protease, pepsin, trypsin, cathepsin, and subtilisin;

[0014] c) Filtration and purification: Filter and separate the aforementioned enzymatic hydrolysate through a microfiltration spiral wound membrane with a pore size of 0.5-2 μm to remove impurities and collect the filtrate, then filter the collected filtrate through an ultrafiltration machine, and collect the fraction with a molecular weight cut-off less than 1000 D to obtain a cistanche peptide extract; and

[0015] d) Concentration and drying: Evaporate and concentrate the aforementioned cistanche peptide extract and then spray dry it to obtain cistanche peptides.

[0016] On the other hand, the present invention provides a process for preparing the aforementioned cistanche peptide composition, which is prepared by compounding each component to obtain the aforementioned cistanche peptide composition. Detailed Description of the Invention

[0018] The present invention provides a cistanche peptide composition, comprising cistanche peptide, extract of pterocarpus marsupium and extract of astragalus membranaceus in specific mass percentages. The applicant has found that when cistanche peptide is combined with other specific components required in this application in a certain mass percentage, the formed composition can enable the active ingredients in cistanche peptide to be more fully absorbed by the human body, so as to achieve better curative effects when applied to neurodegenerative diseases.

[0019] Specifically, cistanche peptide is a small molecule active peptide extracted from cistanche deserticola, with a small molecular weight and being easy to be absorbed by the human body. Without being bound by theory, the applicant has found that cistanche peptide has significant neuroprotective effects and can directly act on nerve cells through the blood-brain barrier. At the same time, components such as astragalus polysaccharide in the extract of astragalus membranaceus in the composition of this application can regulate the immune system and promote the repair and regeneration of nerve cells, while the extract of pterocarpus marsupium is rich in various antioxidant components, such as flavonoid compounds, etc. These components can enhance the antioxidant capacity of the body, reduce the damage of oxidative stress to cells, and have a certain anti-inflammatory effect. The aforementioned three have a synergistic effect in the composition, can jointly reduce oxidative stress and neuroinflammation, protect nerve cells from damage, and thus delay the progression of neurodegenerative diseases to achieve therapeutic effects.

[0020] Cistanche Peptide Composition

[0021] The present invention provides a cistanche peptide composition, which, by mass percentage, comprises the following components: 15%-20% cistanche peptide, 10%-20% extract of pterocarpus marsupium, and 8%-15% extract of astragalus membranaceus. Preferably, in the aforementioned cistanche peptide composition, by mass percentage, the cistanche peptide accounts for 18-20%, the extract of pterocarpus marsupium accounts for 10-15%, and the extract of astragalus membranaceus accounts for 10-12%.

[0022] In some preferred embodiments, 90% of the peptide molecular weight distribution of the aforementioned cistanche peptide is below 1000 daltons, and more preferably 90% is below 500 daltons. The peptide molecular weight distribution can be detected by any conventional and applicable method, and preferably detected by the peptide molecular weight detection method described below, that is, the method in Appendix A of GB / T 22492-2008.

[0023] The composition of the present invention may also contain other suitable components, especially components beneficial to nerve cells, such as Gastrodia elata extract, Citrus reticulata extract, and Crataegus pinnatifida extract. Since ancient times, Gastrodia elata has been used as traditional Chinese medicine to treat symptoms such as neurasthenia and dizziness. Its extract is rich in active ingredients such as gastrodin, which can improve cerebral blood flow, promote the metabolism of nerve cells, and enhance the stress resistance of nerve cells. Citrus reticulata extract is rich in components such as flavonoids, volatile oils, hesperidin, etc., and has antioxidant and anti-inflammatory effects, which can synergistically reduce oxidative stress and neuroinflammation, thereby protecting nerve cells from damage. Crataegus pinnatifida extract is rich in various antioxidant substances such as vitamin C, vitamin E, and carotene, as well as various active ingredients such as flavonoids and triterpenoid saponins, and has antioxidant and anti-inflammatory effects. In some preferred embodiments, the composition of the present invention further includes 8%-15% by mass of Gastrodia elata extract, 5%-10% by mass of Citrus reticulata extract, and 5%-10% by mass of Crataegus pinnatifida extract. More preferably, by mass percentage, the Gastrodia elata extract accounts for 10-12%, the Citrus reticulata extract accounts for 5-8%, and the Crataegus pinnatifida extract accounts for 5-8%.

[0024] In some preferred embodiments, the aforementioned cistanche peptide composition, by mass percentage, comprises the following components: 18%-20% cistanche peptide, 10%-15% Butea superba extract, 10%-12% Astragalus membranaceus extract, 10%-12% Gastrodia elata extract, 5%-8% Citrus reticulata extract, 5%-8% Crataegus pinnatifida extract, and water.

[0025] In a highly more preferred embodiment, the aforementioned cistanche peptide composition, by mass percentage, comprises the following components: 18% cistanche peptide, 12% Butea superba extract, 10% Astragalus membranaceus extract, 10% Gastrodia elata extract, 5% Citrus reticulata extract, 5% Crataegus pinnatifida extract, and water.

[0026] Application of the cistanche peptide composition

[0027] The present invention provides a traditional Chinese medicine composition comprising an effective amount of the aforementioned cistanche peptide composition. In addition to the aforementioned cistanche peptide composition, the traditional Chinese medicine composition may further contain any other suitable components, such as selecting appropriate components according to specific indications.

[0028] The cistanche peptide composition of the present invention is useful for preparing a medicament for preventing or treating neurodegenerative diseases. The aforementioned neurodegenerative diseases are selected from Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, or multiple sclerosis. Preferably, the aforementioned neurodegenerative disease is Alzheimer's disease or Parkinson's disease, and more preferably Parkinson's disease. Without being bound by theory, the applicant has found that the cistanche peptide composition of the present application acts on multiple pathological links of neurodegenerative diseases through the synergy of the above-mentioned effects such as neuroprotection and repair, antioxidant and anti-inflammatory, and enhancement of immune function. The synergistic effect between these components enables the composition to have a preventive or therapeutic effect in improving the symptoms of neurodegenerative diseases, especially showing a strong therapeutic effect on Parkinson's disease.

[0029] Preparation process

[0030] The cistanche peptide in the present invention can be prepared by any conventional method. Preferably, the present invention provides a process for preparing the aforementioned cistanche peptide, comprising the following steps:

[0031] a) Pretreatment: Select fresh cistanche, after washing, slicing, drying, crush it into granules, and then sieve it through an 80-100 mesh sieve to obtain cistanche crude powder;

[0032] b) Enzymatic hydrolysis: Add the cistanche crude powder to 5-8 times the weight of water, heat it to 80-90 °C and stir evenly. After stirring for 10-20 min, cool it down to 40-50 °C, add a composite enzyme according to 0.5-2% of the substrate concentration, stir and enzymatically hydrolyze for 2-3 h. After the enzymatic hydrolysis is completed, raise the temperature to 90-100 °C and inactivate for 10-20 min. After inactivation and cooling, filter it through a 150-200 mesh filter bag to discharge the filter residue to obtain an enzymatic hydrolysate. The composite enzyme contains at least two enzymes selected from the group consisting of cellulase, pectinase, papain, alkaline protease, neutral protease, pepsin, trypsin, cathepsin, and subtilisin;

[0033] c) Filtration and purification: Filter and separate the enzymatic hydrolysate through a microfiltration spiral wound membrane with a pore size of 0.5 - 2 μm to remove impurities and collect the filtrate. Then filter the collected filtrate through an ultrafiltration machine and collect the fraction with a molecular weight cut-off less than 1000 D to obtain a cistanche peptide extract; and

[0034] d) Concentration and drying: Evaporate and concentrate the cistanche peptide extract and then spray dry it to obtain cistanche peptide.

[0035] Preferably, in the pretreatment of step a) above, an 80-mesh sieve is used.

[0036] Preferably, in the enzymatic hydrolysis in step b) above, the complex enzyme is added at 0.5% of the substrate concentration. More preferably, the complex enzyme comprises cellulase and pectinase, and the ratio of the cellulase to the pectinase in the complex enzyme is 1-2:1, more preferably 1-1.5:1.

[0037] Preferably, in the filtration and purification in step c) above, a microfiltration spiral wound membrane with a pore size of 0.5 - 1 μm, more preferably 0.5 μm, is used for membrane filtration separation to remove impurities.

[0038] Preferably, in the concentration and drying in step d) above, a double-effect concentrator is used to achieve evaporation and concentration. A "double-effect concentrator" refers to an evaporation device with two evaporation and concentration efficiencies or stages. In the first stage (the first effect), the solution is heated and evaporated to generate steam, and these steams are not directly discharged, but are used as the heat source for the second stage (the second effect) to heat and evaporate another batch of solution, thereby achieving the full utilization of secondary steam.

[0039] On the other hand, the present invention provides a preparation process of the cistanche peptide composition, and the cistanche peptide composition is prepared by compounding the components in the foregoing composition.

[0040] Peptide molecular weight detection method

[0041] The cistanche peptides prepared by the present invention can be detected for the peptide content with a molecular weight < 500D by any conventional peptide content detection method, preferably by the method in Appendix A of GB / T22492-2008. This standard is included in the National Standard Full-text Public System, see https: / / openstd.samr.gov.cn / bzgk / gb / newGbInfo?hcno=80B693FE7FFFA345807FF3D1AA185DC9. Specific embodiments

[0042] The examples in this article are intended to illustrate the present invention, but not to limit or otherwise define the scope of the present invention.

[0043] Examples 1-2 are examples of the cistanche peptide composition of the present invention, and Examples 3-4 are comparative examples. Example 5 is the preparation process of the cistanche peptide in Examples 1-2, Example 6 is the preparation process of Examples 1-2, Example 7 is the preparation process of Comparative Examples 3-4, and Example 8 is the experimental data of the application effects of each example and comparative example.

[0044] Examples 1-2 and Comparative Examples 3-4: Composition

[0045] The composition shown in Table 1 is composed of the listed components according to the listed mass percentages.

[0046] Table 1

[0047] Example 1 Example 2 Comparative Example 3 Comparative Example 4 Cistanche peptide 18％ 15％ 0 0 Total glycosides of Cistanche 0 0 18％ 15％ Extract of Butea superba 12％ 15％ 12％ 15％ Astragalus membranaceus extract 10％ 8％ 10％ 8％ Gastrodia elata extract 10％ 8％ 10％ 8％ Pericarpium citri reticulatae extract 5％ 7％ 5％ 7％ Fructus crataegi extract 5％ 7％ 5％ 7％ Water Balance Balance Balance Balance

[0048] Example 5: Preparation process of cistanche peptide in Examples 1 - 2

[0049] a) Pretreatment: Select fresh cistanche, after washing, slicing, drying, crush it into particles, then sieve it through an 80 - mesh sieve to obtain crude cistanche powder;

[0050] b) Enzymatic hydrolysis: Add the aforementioned crude cistanche powder to 5 times its weight of water, heat to 85°C and stir evenly. After stirring for 15 min, cool down to 50°C, add a compound enzyme (cellulase, pectinase) at 0.5% of the substrate concentration. The addition ratio of cellulase and pectinase is 1.2:1, stir and enzymatically hydrolyze for 2 h. After the enzymatic hydrolysis is completed, raise the temperature to 100°C and inactivate for 10 min. After inactivation and cooling, filter through a 200 - mesh filter bag to discharge the filter residue and obtain the enzymatic hydrolysate;

[0051] c) Filtration and purification: Filter and separate the aforementioned enzymatic hydrolysate through a microfiltration spiral - wound membrane with a pore size of 0.5 μm to remove impurities and collect the filtrate. Then filter the collected filtrate through an ultrafilter and collect the fraction with a molecular weight cut - off less than 1000 D to obtain the cistanche peptide extract; and

[0052] d) Concentration and drying: First concentrate the aforementioned cistanche peptide extract through a double - effect concentrator and then spray - dry it to obtain cistanche peptide.

[0053] Example 6: Preparation process of cistanche peptide composition in Examples 1 - 2

[0054] Mix the cistanche peptide, extracts of butea superba, astragalus membranaceus, gastrodia elata, citrus reticulata, hawthorn, and the remaining water with the corresponding contents in Table 1 evenly.

[0055] Among them, the cistanche peptide is prepared by the preparation process of Example 5, and the suppliers of other components are as follows: extract of butea superba, Xi'an Dongchi Biotechnology Co., Ltd.; extract of astragalus membranaceus, Xi'an Ruierli Bio - engineering Co., Ltd.; extract of gastrodia elata, Xi'an Ruierli Bio - engineering Co., Ltd.; extract of citrus reticulata, Xi'an Ruierli Bio - engineering Co., Ltd.; extract of hawthorn, Xi'an Ruierli Bio - engineering Co., Ltd.

[0056] Example 7: Preparation process of total cistanche glycosides composition in Comparative Examples 3 - 4

[0057] Mix the total glycosides of Cistanche deserticola, the extract of Butea superba, the extract of Astragalus membranaceus, the extract of Gastrodia elata, the extract of Citrus reticulata Blanco, the extract of Crataegus pinnatifida, and the balance of water with the corresponding contents in Table 1 thoroughly and evenly. Among them, the supplier of the total glycosides of Cistanche deserticola is Shaanxi Baichuan Biotechnology Co., Ltd., and the suppliers of the other components, namely the extract of Butea superba, the extract of Astragalus membranaceus, the extract of Gastrodia elata, the extract of Citrus reticulata Blanco, and the extract of Crataegus pinnatifida, are the same as those in Example 6.

[0058] Example 8: Experimental data of Examples 1-2 and Comparative Examples 3-4 applied to Parkinson's (PD) model mice

[0059] 1) Experimental subjects: Select 90 healthy adult SPF-grade male C57BL / 6 mice, 9 weeks old, weighing 20-25 g, and place them at room temperature of 21-25 °C.

[0060] 2) Experimental reagents: MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride). This reagent can induce Parkinson's in most animals and is currently the most commonly used agent for constructing a mouse PD model.

[0061] 3) Establishment and grouping of the mouse PD model: Randomly divide the mice among the aforementioned experimental subjects into six groups: a control group, a model group, an Example 1 group, an Example 2 group, a Comparative Example 3 group, and a Comparative Example 4 group, with 15 mice in each group. Mice in the model group, Example 1 group, Example 2 group, Comparative Example 3 group, and Comparative Example 4 group were intraperitoneally injected with MPTP at a dose of 30 mg / kg once a day for 7 consecutive days to prepare a mouse PD model; the control group was intraperitoneally injected with an equal volume of normal saline at the same time.

[0062] 4) Experimental method: Starting from the 8th day of the experiment, that is, after the establishment of the mouse PD model, mice in the Example 1-2 groups were given quantitative gavage of the Cistanche peptide composition prepared in Example 1-2 every day, and mice in the Comparative Example 3-4 groups were given quantitative gavage of the total glycosides of Cistanche composition prepared in Comparative Example 3-4 every day, once a day for 14 consecutive days; the control group and the model group were also intraperitoneally injected with an equal volume of normal saline at the same time.

[0063] 5) Behavioral experiment

[0064] Mice in each group were given 5 times of pole climbing and hanging adaptability behavior training every day for 2 days before the experiment, and then pole climbing and hanging behavioral observations were carried out on the 1st day before the experiment (1 day before the first injection of MPTP), the 7th day (after injection of MPTP), the 14th day (7 days after gavage), and the 21st day (14 days after gavage) to detect their limb movement coordination.

[0065] Pole climbing experiment

[0066] Fix a plastic ball with a diameter of 2.5 cm at the top of a wooden rod that is 60 cm long and 1 cm in diameter. Wrap two layers of gauze around the wooden rod to prevent slipping. Place the mouse on top of the ball and record the following three times: (1) the time it takes for the mouse to climb the upper part of the rod length; (2) the time it takes for the mouse to climb the lower part of the rod length; (3) the time it takes for the mouse to climb the full length of the rod. Score according to the following criteria: If a certain action above is completed within 3 seconds, record 3 points; if completed within 6 seconds, record 2 points; if it exceeds 6 seconds, record 1 point. The sum of the scores of the three times is the score of the mouse pole climbing experiment.

[0067] Hanging experiment

[0068] Suspend the mouse's two front limbs on a horizontal electric wire. If the mouse grabs the wire with its two hind limbs, record 3 points; if it grabs the wire with only one hind limb, record 2 points; if it cannot grab with both hind limbs, record 1 point; if the mouse falls, record 0 points.

[0069] 6) Statistical processing: Use SPSS software for statistical processing. The data is expressed as mean ± standard deviation (x±s), and a t-test is used for significant difference analysis. P<0.01 indicates a highly significant difference; P<0.05 indicates a significant difference.

[0070] 7) Experimental results

[0071] The scoring results of the mouse pole climbing experiment are shown in Table 2.

[0072] Table 2

[0073] Group 1 day before the experiment Day 7 Day 14 Day 21 Control group 6.00±1.23 6.13±1.14 6.20±1.09 6.27±1.05 Model group 6.07±1.18 2.00±0.82 2.13±0.74 2.20±0.71 Example 1 group 6.00±1.20 2.13±0.87 4.87±1.04 5.67±0.98 Example 2 group 6.13±1.15 2.07±0.93 4.20±1.12 4.93±1.03 Comparative Example 3 group 6.00±1.22 2.00±0.88 3.53±1.21 3.93±1.14 Comparative Example 4 group 6.07±1.19 1.93±0.96 3.00±1.30 3.27±1.23

[0074] One day before the experiment, observe the mouse's pole climbing behavior. The scores of the mouse pole climbing experiment in the model group, Example 1-2 groups, and Comparative Example 3-4 groups have no differences compared with the control group (P>0.05, P>0.05, P>0.05, P>0.05, P>0.05); on the 7th, 14th, and 21st days of the experiment, observe the mouse's pole climbing behavior in the model group. Compared with the control group, the experimental scores are significantly reduced, with highly significant differences (P<0.01, P<0.01, P<0.01); on the 14th and 21st days of the experiment, the experimental scores of the Example 1-2 groups and Comparative Example 3-4 groups are significantly increased, with significant differences compared with the model group (P<0.05, P<0.05, P<0.05, P<0.05); on the 14th and 21st days of the experiment, the Comparative Example 3-4 groups are compared with the Example 1 group and the Example 2 group respectively, and their experimental scores are lower, with significant differences (P<0.05, P<0.05, P<0.05).

[0075] The scoring results of the mouse hanging experiment are shown in Table 3.

[0076] Table 3

[0077] Group 1 day before the experiment Day 7 Day 14 Day 21 Normal control group 2.53±0.51 2.60±0.49 2.67±0.46 2.73±0.43 Model group 2.60±0.48 0.67±0.49 0.73±0.46 0.73±0.46 Example 1 group 2.53±0.50 0.73±0.50 1.87±0.61 2.33±0.57 Example 2 group 2.60±0.47 0.67±0.49 1.53±0.68 1.93±0.63 Comparative Example 3 group 2.53±0.51 0.60±0.50 1.27±0.71 1.47±0.64 Comparative Example 4 group 2.60±0.48 0.53±0.51 0.93±0.71 1.07±0.64

[0078] One day before the experiment, the hanging behavior of the mice was observed. There were no differences in the hanging experiment scores of the mice in the model group, Example 1-2 groups, and Comparative Example 3-4 groups compared with the control group (P>0.05, P>0.05, P>0.05, P>0.05, P>0.05); on the 7th, 14th, and 21st days of the experiment, the hanging behavior of the mice in the model group was observed. Compared with the control group, the experimental scores were significantly decreased, and the differences were extremely significant (P<0.01, P<0.01, P<0.01); on the 14th and 21st days of the experiment, the hanging experiment scores of the mice in Example 1-2 groups and Comparative Example 3-4 groups were significantly increased, and the differences were significant compared with the model group (P<0.05, P<0.05, P<0.05, P<0.05); on the 14th and 21st days of the experiment, the experimental scores of Comparative Example 3-4 groups were lower compared with Example 1 group and Example 2 group respectively, and the differences were significant (P<0.05, P<0.05, P<0.05).

[0079] According to the experimental results of the above pole climbing experiment and hanging experiment of the mice, it can be seen that after the mice in the model group were injected with MPTP, their limb movement coordination ability was significantly decreased, indicating that the construction of the mouse Parkinson's disease model was successful. After the mice in Example 1 group and Example 2 group were intragastrically administered with the cistanche peptide composition of the present invention, their limb movement coordination ability was restored to a certain extent (and the restoration effect of Example 1 group was better than that of Example 2 group), which was significantly better than that of Comparative Example 3 group and Comparative Example 4 group. Therefore, the cistanche peptide composition of the embodiments of the present invention exhibits excellent effects in improving the mouse Parkinson's disease model.

[0080] Although specific embodiments of the present invention have been described and illustrated, those skilled in the art should understand that various other changes and modifications can be made without departing from the spirit and scope of the present invention. Therefore, all such changes and modifications fall within the scope of the claims of this application.

[0081] Unless otherwise specified, all percentages, ratios, and proportions are calculated based on the weight of the total composition. Unless otherwise specified, all temperatures are in degrees Celsius (°C). All component or composition levels refer to the active levels of the components or compositions, excluding impurities that may be present in commercially available sources, such as residual solvents or by-products.

[0082] It should be understood that each upper numerical limit given in this specification includes the lower numerical values, as if these lower numerical values were explicitly written in the application. Each lower numerical limit given in this specification includes the higher numerical values, as if these higher numerical values were explicitly written in the application. Each numerical range given in this specification includes the narrower numerical ranges falling within that numerical range, as if these narrower numerical ranges were explicitly written in the application.

[0083] The numerical values and dimensions disclosed herein should not be construed as strictly limited to the exact values recited. Instead, each such numerical value and dimension is intended to represent the recited value and a functionally equivalent range around that value. For example, a dimension disclosed as "40 mm" is intended to mean "about 40 mm".

[0084] As used herein, the term "mixing" refers to adding ingredients together and achieving homogeneity, and the term "mixture" refers to a homogeneous mixture of ingredients.

[0085] In this application, when a composition is "substantially free of" a particular ingredient, it means that the composition contains less than a trace amount, or less than 0.1%, or less than 0.01%, or less than 0.001%, by mass of the composition, of the particular ingredient.

[0086] In this application, unless the order is specifically defined, the terms "a)", "b)", "c)", etc. used to refer to steps in a preparation method do not denote an order. For example, step a) may occur before step b) or after step b), or steps b) and c) may occur simultaneously, or step b) may occur after step c).

[0087] As used herein, the terms "comprising", "including", "containing", "comprises", "includes", "contains", "comprised of", "include", and "contains" are intended to be non-limiting, i.e., other ingredients and other steps may be added without affecting the final result.

Claims

1. A cistanche peptide composition, by mass percentage, comprises the following components: 15%-20% cistanche peptide, 10%-20% extract of Eurycoma longifolia, and 8%-15% extract of Astragalus membranaceus.

2. The cistanche peptide composition according to claim 1, wherein 90% of the peptide molecular weight distribution of the cistanche peptide is below 500 daltons.

3. The cistanche peptide composition according to claim 1, further comprises 8%-15% extract of Gastrodia elata, 5%-10% extract of Citrus reticulata Blanco and 5%-10% extract of Crataegus pinnatifida Bunge by mass percentage.

4. The cistanche peptide composition according to claim 3, by mass percentage, comprises the following components: 18%-20% cistanche peptide, 10%-15% extract of Eurycoma longifolia, 10%-12% extract of Astragalus membranaceus, 10%-12% extract of Gastrodia elata, 5%-8% extract of Citrus reticulata Blanco, 5%-8% extract of Crataegus pinnatifida Bunge, and water.

5. A traditional Chinese medicine composition comprises an effective amount of the cistanche peptide composition according to any one of claims 1-4.

6. Use of the cistanche peptide composition according to any one of claims 1-4 for the preparation of a medicament for preventing or treating neurodegenerative diseases.

7. The use according to claim 6, wherein the neurodegenerative disease is Parkinson's disease.

8. A process for preparing the cistanche peptide according to any one of claims 1-4, comprises the following steps: a) Pretreatment: Select fresh cistanche, after cleaning, slicing, drying, crush into particles, then sieve with 80-100 mesh sieve to obtain cistanche coarse powder; b) Enzymolysis: Add the cistanche coarse powder into 5-8 times by weight of water, heat to 80-90 °C and stir evenly, after stirring for 10-20 min, cool down to 40-50 °C, add compound enzyme according to 0.5-2% of the substrate concentration, stir and enzymolyze for 2-3 h, after the enzymolysis ends, heat up to 90-100 °C, inactivate for 10-20 min, after inactivation and cooling, filter with 150-200 mesh filter bag, discharge the filter residue to obtain enzymolysis liquid, the compound enzyme comprises at least two enzymes selected from the group consisting of cellulase, pectinase, papain, alkaline protease, neutral protease, pepsin, trypsin, cathepsin and subtilisin; c) Filtration and purification: Filter the enzymolysis liquid through a microfiltration spiral wound membrane with a pore size of 0.5-2 μm to separate and remove impurities, collect the filtrate, then filter the collected filtrate through an ultrafiltration machine, collect the fraction with a molecular weight cut-off less than 1000 D to obtain cistanche peptide extract; and d) Concentration and drying: First evaporate and concentrate the cistanche peptide extract and then spray dry it to obtain cistanche peptide.

9. The process for preparing cistanche peptide according to claim 8, wherein the compound enzyme comprises cellulase and pectinase, and the ratio of the cellulase and the pectinase in the compound enzyme is 1-2:1, preferably 1-1.5:

1.

10. A process for preparing the cistanche peptide composition according to any one of claims 1-4, the cistanche peptide composition is prepared by compounding each component.