Traditional Chinese medicine monomer or composition and application thereof

Abstract

According to the invention, two traditional Chinese medicine monomers, namely Eupalinolide K and N-p-coumaroyl-5-hydroxytryptamine (N-(p-Coumaroyl) Sorotonin), which have a relieving effect on the Parkinson's disease are screened out for the first time, and a medicine or a pharmaceutical composition containing the two traditional Chinese medicine monomers and an application of the medicine or the pharmaceutical composition are provided. Molecular mechanisms of the two traditional Chinese medicine monomers for relieving the PD are studied, the characteristics of the traditional Chinese medicine monomers are predicted and verified by utilizing a prediction tool and an animal model experiment, and the result shows that lindley eupatorium lactone K can possibly penetrate through a blood brain barrier to play a role and has an application prospect in preparation of medicines for treating the PD; a verification result shows that the lindley eupatorium lactone K and N-p-coumaroyl-5-hydroxytryptamine can provide a new action target and a new treatment strategy for PD treatment.

Description

Technical Field

[0001] The present invention relates to the fields of traditional Chinese medicine and neurological disease treatment, and in particular to a traditional Chinese medicine monomer or composition and use thereof in preparing a medicine for treating Parkinson's disease. Background Art

[0002] Parkinson's disease (PD) is a typical neurodegenerative disorder characterized by progressive degeneration of dopaminergic neurons in the substantia nigra and abnormal aggregation of α-synuclein. Clinically, core motor symptoms include bradykinesia, tremor, and muscle rigidity, primarily due to a significant deficiency of dopamine in the striatum.

[0003] Currently, drug therapy remains the main treatment for PD. However, most existing drug therapies can only relieve symptoms but cannot prevent disease progression, and long-term use often causes serious side effects. After the advent of levodopa in the 1960s, dopamine replacement therapy became the core strategy for PD treatment. Although long-term use of levodopa can effectively improve motor symptoms, the efficacy gradually decreases and motor complications such as dyskinesia occur, which greatly limits its long-term application. Dopamine receptor agonists (such as rotigotine transdermal patch) and monoamine oxidase B inhibitors (such as rasagiline) can delay the use of levodopa to a certain extent, but they can cause non-motor side effects such as hallucinations and impulse control disorders.

[0004] In recent years, traditional Chinese medicine (TCM) and its monomers (single active ingredients isolated from traditional Chinese medicine) have become a hot topic in PD treatment research due to their multi-target effects, low toxicity, and neuroprotective potential. These compounds exhibit unique therapeutic advantages through mechanisms such as regulating oxidative stress, inhibiting α-synuclein aggregation, and ameliorating mitochondrial dysfunction.

[0005] Therefore, there is an urgent need in this field to discover a Chinese medicine monomer or composition that can be used to prepare a drug for treating Parkinson's disease, and its use in treating PD, in order to provide new targets and treatment strategies for PD treatment. Summary of the Invention

[0006] The purpose of the present invention is to provide a traditional Chinese medicine monomer or composition and its use in preparing a medicine for treating Parkinson's disease.

[0007] The first aspect of the present invention provides a medicine or a pharmaceutical composition, comprising:

[0008] (1) Eupalinolide K;

[0009] (2) N-(p-Coumaroyl)Serotonin;

[0010] (3) (Z)-Guggulsterone;

[0011] or a combination thereof.

[0012] In another preferred embodiment, the drug or drug composition is a drug monomer.

[0013] In another preferred embodiment, the drug or pharmaceutical composition is yamalide K.

[0014] In another preferred embodiment, the drug or pharmaceutical composition is N-p-coumaryl-5-hydroxytryptamine.

[0015] In another preferred embodiment, the drug or pharmaceutical composition is (Z)-bisabolsterone.

[0016] In another preferred embodiment, the drug or pharmaceutical composition further comprises a pharmaceutically acceptable carrier, diluent or excipient.

[0017] In another preferred embodiment, the drug or pharmaceutical composition comprises: yamachulide K, N-p-coumaryl-5-hydroxytryptamine, or a combination thereof.

[0018] In another preferred embodiment, the drug or pharmaceutical composition comprises:

[0019] (a) a therapeutically and / or prophylactically effective amount of yamadrolactone K, N-p-coumaryl-5-hydroxytryptamine, (Z)-bisabolsterone, or a combination thereof; and

[0020] (b) a pharmaceutically acceptable carrier, diluent or excipient.

[0021] In another preferred embodiment, the drug or pharmaceutical composition further comprises:

[0022] (4)Dehydrocorydaline nitrate;

[0023] (5)11-Keto-beta-boswellic acid;

[0024] (6) Loganin;

[0025] (7) Butein;

[0026] (8) Curcumin;

[0027] (9)Iristectorigenin B;

[0028] (10)Arachidonic acid;

[0029] (11) Evodiamine;

[0030] (12)Acacetin;

[0031] (13) Kushenol I;

[0032] or a combination thereof.

[0033] In another preferred embodiment, the drug or pharmaceutical composition further comprises:

[0034] (c) a therapeutically and / or prophylactically effective amount of Dehydrocorydaline nitrate, 11-Keto-beta-boswellic acid, Loganin, Butein, Curcumin, Iristectorigenin B, Arachidonic acid, Evodiamine, Acacetin, Kushenol I, or a combination thereof.

[0035] The second aspect of the present invention provides a use of the drug or pharmaceutical composition according to the first aspect of the present invention for preparing a drug for treating and / or preventing neurodegenerative diseases.

[0036] In another preferred embodiment, the drug or pharmaceutical composition is selected from the following group: yamachulide K, N-p-coumaryl-5-hydroxytryptamine, or a combination thereof.

[0037] In another preferred embodiment, the drug or pharmaceutical composition is yamalide K.

[0038] In another preferred embodiment, the neurodegenerative disease includes Parkinson's disease.

[0039] In another preferred embodiment, the use also includes: for preparing a drug for treating and / or preventing Parkinson's disease.

[0040] In another preferred embodiment, the use also includes: (a) for preparing a drug for reducing and / or inhibiting the damage or death of nerve cells, and / or (b) for preparing a drug for promoting the survival of nerve cells.

[0041] The third aspect of the present invention provides a medicine kit comprising:

[0042] (A) a first medicine kit, and the drug or pharmaceutical composition according to the first aspect of the present invention located in the first medicine kit;

[0043] (B) a second drug kit, and other drugs for treating neurodegenerative diseases in the second drug kit.

[0044] In another preferred embodiment, the neurodegenerative disease includes Parkinson's disease.

[0045] In another preferred embodiment, the medicine kit further comprises a label or instructions.

[0046] In another preferred embodiment, the label or instructions indicate that the drug kit is used to treat and / or prevent neurodegenerative diseases.

[0047] In another preferred embodiment, the label or instructions indicate that the drug kit is used to treat and / or prevent Parkinson's disease.

[0048] In another preferred embodiment, the drug or pharmaceutical composition is selected from the following group: yamachulide K, N-p-coumaryl-5-hydroxytryptamine, (Z)-bisabolsterone, or a combination thereof.

[0049] In another preferred embodiment, the drug or pharmaceutical composition is selected from the following group: yamachulide K, N-p-coumaryl-5-hydroxytryptamine, or a combination thereof.

[0050] In another preferred embodiment, the drug or pharmaceutical composition is yamalide K.

[0051] A fourth aspect of the present invention provides a method for reducing and / or inhibiting nerve cell damage and / or death, comprising the steps of:

[0052] (a) providing a nerve cell;

[0053] (b) incubating the drug or pharmaceutical composition according to the first aspect of the present invention with the nerve cells, thereby reducing and / or inhibiting damage and / or death of the nerve cells.

[0054] In another preferred embodiment, the neural cells include: neural crest cells and neuronal cells.

[0055] In another preferred embodiment, the neural crest cells include neuroblastoma cells.

[0056] In another preferred embodiment, the neuronal cells include dopaminergic neuronal cells.

[0057] In another preferred embodiment, the neural cells are derived from humans or non-human mammals.

[0058] In another preferred embodiment, the non-human mammals include: mice and rats.

[0059] In another preferred embodiment, the neuroblastoma cells are human neuroblastoma cells (SH-SY5Y).

[0060] In another preferred embodiment, the neuroblastoma cells are damaged neuroblastoma cells.

[0061] In another preferred embodiment, the neuroblastoma cells are MPP + Induced damage in human neuroblastoma cells.

[0062] In another preferred embodiment, the dopaminergic neuron cells are mouse dopaminergic neuron cells.

[0063] In another preferred embodiment, the dopaminergic neuron cells are mouse tyrosine hydroxylase (TH)-positive dopaminergic neuron cells.

[0064] In another preferred embodiment, the dopaminergic neuron cells are MPTP-induced damaged mouse tyrosine hydroxylase (TH)-positive dopaminergic neuron cells.

[0065] In another preferred embodiment, the drug or pharmaceutical composition is combined with the MPP + Induced damage to human neuroblastoma cells for incubation, thereby reducing and / or inhibiting the MPP + Induces damage and / or death in damaged human neuroblastoma cells.

[0066] In another preferred embodiment, the drug or pharmaceutical composition is incubated with the MPTP-induced damaged mouse tyrosine hydroxylase (TH)-positive dopaminergic neuronal cells, thereby reducing and / or inhibiting the damage and / or death of the MPTP-induced damaged mouse tyrosine hydroxylase (TH)-positive dopaminergic neuronal cells.

[0067] In another preferred embodiment, the drug or pharmaceutical composition is selected from the following group:

[0068] (1) Eupalinolide K;

[0069] (2) N-(p-Coumaroyl)Serotonin;

[0070] (3) (Z)-Guggulsterone;

[0071] (4)Dehydrocorydaline nitrate;

[0072] (5)11-Keto-beta-boswellic acid;

[0073] (6) Loganin;

[0074] (7) Butein;

[0075] (8) Curcumin;

[0076] (9)Iristectorigenin B;

[0077] (10)Arachidonic acid;

[0078] (11) Evodiamine;

[0079] (12)Acacetin;

[0080] (13) Kushenol I;

[0081] or a combination thereof.

[0082] In another preferred embodiment, the drug or pharmaceutical composition is selected from the following group:

[0083] (1) Yemachalide K;

[0084] (2) N-p-coumaryl-5-hydroxytryptamine;

[0085] (3) (Z)-bisabolsterone;

[0086] or a combination thereof.

[0087] In another preferred embodiment, the drug or pharmaceutical composition is selected from the following group: yamachulide K, N-p-coumaryl-5-hydroxytryptamine.

[0088] In another preferred embodiment, the drug or drug composition is a drug monomer.

[0089] In another preferred embodiment, the drug or pharmaceutical composition is yamalide K.

[0090] In another preferred embodiment, the drug or pharmaceutical composition is N-p-coumaryl-5-hydroxytryptamine.

[0091] In another preferred embodiment, the drug or pharmaceutical composition is (Z)-bisabolsterone.

[0092] In another preferred embodiment, the dosage of the drug or pharmaceutical composition is ≥1 mg / kg, such as 3 mg / kg, 10 mg / kg, or 20 mg / kg.

[0093] In another preferred embodiment, the concentration of the drug or pharmaceutical composition is ≥5 μM.

[0094] In another preferred embodiment, the concentration of the drug or pharmaceutical composition is selected from the following group: 5 μM, 10 μM, 20 μM.

[0095] In another preferred embodiment, the nerve cells are incubated with yamachulide K, and the concentration of yamachulide K is ≥5 μM, preferably 10 μM.

[0096] In another preferred embodiment, N-p-coumaryl-5-hydroxytryptamine is incubated with the nerve cells, and the concentration of the N-p-coumaryl-5-hydroxytryptamine is ≥10 μM, such as 20 μM.

[0097] In another preferred embodiment, (Z)-bisabolsterone is incubated with the nerve cells, and the concentration of the (Z)-bisabolsterone is ≥10 μM, such as 20 μM.

[0098] In another preferred embodiment, the reducing and / or inhibiting the damage and / or death of the nerve cells includes: increasing the activity of nerve cells and / or increasing the proliferation ability of nerve cells.

[0099] In another preferred embodiment, the improving the proliferation ability of nerve cells includes: increasing the number of nerve cells and increasing the proportion of nerve cells.

[0100] In another preferred embodiment, the reducing and / or inhibiting the damage and / or death of the nerve cells includes: increasing the activity and / or number of nerve cells.

[0101] In another preferred embodiment, the nerve cells are incubated with the yamachulide K, and the activity and / or number of the nerve cells are significantly increased.

[0102] In another preferred embodiment, when the yamachulide K is incubated with the nerve cells, the activity of the nerve cells is significantly (p<0.05) increased.

[0103] In another preferred embodiment, the "significantly improved activity" means that the cell activity A1 of the induced damaged human neuroblastoma cells incubated with Nomadolactone K, compared with the cell activity A0 of the induced damaged human neuroblastoma cells, satisfies A1 / A0≥140%, and p<0.05, such as 142%.

[0104] In another preferred embodiment, the nerve cells are incubated with the yamachulide K, and the number of the nerve cells is significantly (p<0.05) increased.

[0105] In another preferred embodiment, the N-p-coumaryl-5-hydroxytryptamine is incubated with the nerve cells, and the activity and / or number of the nerve cells are significantly increased.

[0106] In another preferred embodiment, when the N-p-coumaryl-5-hydroxytryptamine is incubated with the nerve cells, the activity of the nerve cells is significantly (p<0.05) increased.

[0107] In another preferred embodiment, the "significantly improved activity" means that the cell activity A2 of the induced damaged human neuroblastoma cells incubated with N-p-coumaroyl-5-hydroxytryptamine, compared with the cell activity A0 of the induced damaged human neuroblastoma cells, satisfies A2 / A0≥116%, and p<0.05, such as 118%.

[0108] In another preferred embodiment, when the N-p-coumaryl-5-hydroxytryptamine is incubated with the nerve cells, the number of the nerve cells is significantly (p<0.05) increased.

[0109] In another preferred embodiment, the "significantly increased number" means that the number B1 of tyrosine hydroxylase (TH)-positive dopaminergic neuronal cells in mice incubated with N-p-coumaroyl-5-hydroxytryptamine, compared with the number B0 of tyrosine hydroxylase (TH)-positive dopaminergic neuronal cells in mice induced to be injured, satisfies B1 / B0≥130%, and p<0.05, such as 135%.

[0110] In another preferred embodiment, the (Z)-bisabolsterone is incubated with the nerve cells, and the activity of the nerve cells is significantly (p<0.05) increased.

[0111] In another preferred embodiment, the "significantly improved activity" means that the cell activity A3 of the induced damaged human neuroblastoma cells incubated with (Z)-bisabolsterone, compared with the cell activity A0 of the induced damaged human neuroblastoma cells, satisfies A3 / A0≥111%, and p<0.05, such as 116%.

[0112] In another preferred embodiment, the method is non-therapeutic or non-diagnostic.

[0113] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0114] Figure 1 2 mM MPP is shown + The changes in the survival rate of SH-SY5Y cells after treatment with 165 Chinese herbal medicine monomers under induction, *P<0.05, not significant (ns)P≥0.05.

[0115] Figure 2 3 mM MPP is shown + Induced, given Figure 1The screened active monomer compounds (Z)-bisabolsterone, yamadrin K and N-p-coumaryl-5-hydroxytryptamine were further tested for MPP + Changes in SH-SY5Y cell survival rate after treatment, *P<0.05, not significant (ns) P≥0.05.

[0116] Figure 3 Shows the effects of different concentrations of (Z)-bisabolsterone, yamadrin K, and N-p-coumaryl-5-hydroxytryptamine on MPP under another administration method. + The changes in SH-SY5Y cell viability induced by ELISA were *P<0.05, not significant (ns) P≥0.05.

[0117] Figure 4 The pharmacokinetics of three traditional Chinese medicine monomers, (Z)-bisabolsterone, yamachulide K and N-p-coumaryl-5-hydroxytryptamine, predicted by SwissADME are shown.

[0118] Figure 5 The pharmacokinetics of three traditional Chinese medicine monomers, (Z)-bisabolsterone, yamachulide K and N-p-coumaryl-5-hydroxytryptamine, predicted by admetSAR 3.0 are shown.

[0119] Figure 6 The pkCSM prediction of the pharmacokinetics of three traditional Chinese medicine monomers, (Z)-bisabolsterone, yamachulide K and N-p-coumaryl-5-hydroxytryptamine, is shown.

[0120] Figure 7 This study shows that N-p-coumaryl-5-hydroxytryptamine has a neuroprotective effect in the MPTP mouse model. A: Immunofluorescence staining of TH in the substantia nigra of the midbrain + Dopaminergic neurons (scale: 100 μm) were counted using Image J; B: Open field test trajectory of mice and total walking distance and percentage of time spent in the central grid; CD: Behavioral tests of the fatigue rotarod test and the climbing pole test. DETAILED DESCRIPTION

[0121] Through extensive and in-depth research, the inventors have, for the first time, identified two traditional Chinese medicine monomers with PD-alleviating effects: yemachulide K and N-p-coumaroyl-5-hydroxytryptamine. Furthermore, the inventors further investigated the molecular mechanisms underlying the PD-alleviating effects of these two traditional Chinese medicine monomers. Using predictive tools and animal model experiments, they predicted and validated the properties of these monomers, finding that yemachulide K may be able to cross the blood-brain barrier and has potential application in the development of PD treatment drugs. Validation results indicate that yemachulide K and N-p-coumaroyl-5-hydroxytryptamine may provide novel targets and therapeutic strategies for PD treatment. This work, based on these findings, led to the completion of the present invention.

[0122] It should be understood that the specific methods and experimental conditions of the present invention are described below in various levels of detail to provide a more detailed understanding of the present invention. The following provides definitions of certain terms used in this specification. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs.

[0123] the term

[0124] Where a numerical range is provided, it is understood that every intermediate integer of that value, every tenth of each intermediate integer of that value, between the upper and lower limits of that range, and any other intermediate values ​​in the specified range are encompassed within the present invention, unless the context clearly indicates otherwise. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the present invention, subject to any express exclusions in the specified range. For example, "1 to 50" includes "2 to 25," "5 to 20," "25 to 50," "1 to 10," etc.

[0125] As used herein, the terms "comprising" or "including" may be open, semi-closed, or closed. In other words, the terms also include "consisting essentially of" or "consisting of."

[0126] As used herein, the term "and / or" refers to and encompasses any and all possible combinations of one or more of the associated listed items.

[0127] As used herein, the term "significant" means that in a hypothesis test, the observed effect (such as the difference between the experimental group and the control group) is unlikely to be caused by random error alone. A hypothesis test includes: the null hypothesis (H0), which assumes that the observed effect does not exist (such as there is no difference between the experimental group and the control group); the p-value, which is the probability of observing the current or more extreme effect when H0 is true; and the significance threshold (α). The significance threshold is usually used to determine whether the hypothesis test is significant. Generally speaking, the significance threshold is 0.05. If the p-value ≤ α, H0 is rejected, that is, the observed effect exists, and the result is called "significant". In this article, the significance threshold is 0.05.

[0128] As used herein, the terms "Parkinson's disease" and "PD" can be used interchangeably. It is a common neurodegenerative disease whose main pathological feature is the degeneration and death of dopaminergic neurons in the substantia nigra, resulting in a significant decrease in striatal dopamine content, which further manifests as motor dysfunction in patients.

[0129] As used herein, the term "(Z)-Guggulsterone" is an active ingredient of Commiphora mukul, a traditional Indian Ayurvedic medicinal plant. (Z)-Guggulsterone has been reported to be useful in treating and / or preventing neurological diseases, including reducing neuroinflammation and synaptic deficits, restoring cognition, and reversing neuropathological disorders and memory impairment.

[0130] As used herein, the term "MPP + "Induction" and "MPTP induction" are two commonly used Parkinson's disease modeling methods. Other Parkinson's disease modeling methods are well known to those skilled in the art. In this article, MPP + Inducing the construction of an in vitro Parkinson's disease cell (PD cell) model, using MPTP to induce the construction of a Parkinson's disease animal model (PD animal model, such as a PD mouse model, and also a subacute PD mouse model). + The induced PD cell model showed decreased neuronal cell survival / activity, and the MPTP-induced PD animal model showed decreased tyrosine hydroxylase (TH)-positive dopaminergic neuron survival / activity.

[0131] Traditional Chinese medicine monomer of the present invention

[0132] In this paper, two traditional Chinese medicine monomers that have not been reported in PD were screened using the CCK-8 (Cell Counting Kit-8) method.

[0133] As used herein, the term "Eupalinolide K" refers to a sesquiterpene lactone compound derived from Eupalinolide, a class of STAT3 inhibitors. The STAT3 pathway plays an important role in tissue repair, wound healing, and immune response; overactivation of this pathway is associated with a variety of diseases, including cancer. Therefore, Eupalinolide K has the potential to be used as a tumor therapeutic agent.

[0134] As used herein, the term "N-(p-Coumaroyl) Serotonin" refers to a polyphenolic compound isolated from safflower seeds with multi-target pharmacological activity. N-(p-Coumaroyl) Serotonin has potential applications in kidney injury by regulating oxidative stress, inflammation, and apoptosis.

[0135] Drug or pharmaceutical composition and method of administration

[0136] The pharmaceutical composition of the present invention comprises a safe and effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably 10-1000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0137] A typical formulation is prepared by mixing the Chinese herbal medicine monomer or combination of Chinese herbal medicine monomers described in the first aspect of the present invention with a carrier, diluent, or excipient. Suitable carriers, diluents, or excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or swellable polymers, hydrophilic or hydrophobic substances, gelatin, oils, solvents, water, and the like.

[0138] The specific carrier, diluent or excipient used will be determined according to the method of use and purpose of the Chinese medicine monomer of the present invention. Generally, solvents are selected based on solvents that those skilled in the art believe can be safely and effectively administered to mammals. Generally speaking, safe solvents are non-toxic aqueous solvents, such as water, and other non-toxic solvents that are soluble in water or miscible with water. Suitable aqueous solvents include one or more of water, ethanol, propylene glycol, polyethylene glycol (such as PEG400, PEG300) etc. The formula may also include one or more sustained-release agents, tranquilizers, surfactants, lubricants, emulsifiers, suspoemulsions, preservatives, antioxidants, sunscreens, glidants, processing aids, colorants, sweeteners, flavorings, flavorings or other known additives so that the medicine is manufactured or used in an acceptable form.

[0139] The term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. Among them, "compatibility" means that the components in the composition can be mixed with the Chinese medicine monomer or Chinese medicine monomer combination of the present invention and with each other without significantly reducing the efficacy of the compound. Pharmaceutically acceptable carriers include: cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0140] When the Chinese medicine monomer of the present invention is used in combination with at least one or other drugs, the two drugs or multiple drugs can be used separately or in combination, and preferably, are administered in the form of a pharmaceutical composition. The Chinese medicine monomer or pharmaceutical composition of the present invention can be administered orally, by inhalation, parenterally, orally, sublingually, rectum, vaginally, by patch, pump or transdermally, and is accordingly configured into a pharmaceutical composition. Parenteral administration includes, for example, intravenous, intraarterial, subcutaneous, nasal, intrapulmonary, rectal, and topical methods. These pharmaceutical compositions may also contain one or more sustained-release agents, tranquilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, sunscreens, glidants, processing aids, colorants, sweeteners, flavorings, flavorings or other known additives, so that the pharmaceutical composition is manufactured or used in an acceptable form.

[0141] The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

[0142] There is no particular limitation on the administration of the drug or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0143] The medicament of the present invention is preferably administered orally. Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents.

[0144] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes. If desired, the active compound can also be microencapsulated with one or more of the above-mentioned excipients.

[0145] In the case of capsules and tablets, the dosage form may also include a buffering agent. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using lactose and polyethylene glycol as excipients.

[0146] Other materials known in the art include liposomes. Liposomes are composed of various phospholipids, such as cholesterol, stearylamine, or phosphatidylcholine.

[0147] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, or mixtures thereof.

[0148] Besides the above-mentioned inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, and perfumes.

[0149] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, tragacanth, aluminum methoxide and agar-agar, or mixtures of these substances.

[0150] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0151] Dosage forms for topical administration of the compounds of this invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0152] The medicaments or pharmaceutical compositions of the present invention may be administered using other topical formulations, including creams, powders, sprays, suppositories, and inhalants. The medicaments may be mixed under sterile conditions with a pharmaceutically acceptable excipient, diluent, or carrier, and any desired preservative, buffer, or propellant. Ophthalmic formulations, ophthalmic ointments, powders, and solutions are also within the scope of the present invention.

[0153] The medicaments or pharmaceutical compositions of the present invention can be administered nasally or by inhalation, allowing convenient delivery of the drug in the form of a solution, dry powder, or suspension from a pump spray system. The sealed pump spray container is an integral dispensing device and, when the dosage form includes an aerosol dispenser, contains a compressed gas, including but not limited to heptafluoroalkane, carbon dioxide, dichlorodifluoromethane, and the like.

[0154] In the case of suppositories, the compound in the form of a suppository is generally solid at room temperature, melts at body temperature, and can be applied topically to the vagina, rectum, and urethra. The materials commonly used to make carriers for suppositories include, but are not limited to, mixtures of polyethylene glycols, glycerinated gelatin, cocoa butter, hydrogenated vegetable oils, and the like.

[0155] Combination and administration of drugs or pharmaceutical compositions

[0156] The drug of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as anti-neurological disease drugs, such as anti-PD drugs).

[0157] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

[0158] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is administered to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective or prophylactic dosage. For a 60 kg body weight person, the daily dosage is generally 1-2000 mg, preferably 50-1000 mg. The actual dosage, rate of administration, and time of administration depend on the nature and severity of the disease / disorder and the specific drug or pharmaceutical composition being administered. Prescribing a treatment, such as determining the dosage, is within the skill of a skilled physician and generally takes into account the disease / disorder to be treated, the individual subject's condition, the site of administration, the method of administration, and other factors.

[0159] Preferably, the subject is a human or a non-human mammal; the non-human mammal includes rats, mice, and monkeys.

[0160] Use of the drug or pharmaceutical composition

[0161] The present invention provides uses of the drug or pharmaceutical composition for preparing a medicament for treating and / or preventing neurodegenerative diseases. Preferably, the drug or pharmaceutical composition is selected from the group consisting of yamachulide K, N-p-coumaroyl-5-hydroxytryptamine, or a combination thereof. Preferably, the drug or pharmaceutical composition is yamachulide K.

[0162] The use of the drug or pharmaceutical composition of the present invention also includes the preparation of a drug for treating and / or preventing Parkinson's disease. Preferably, the drug or pharmaceutical composition is selected from the group consisting of yamachulide K, N-p-coumaryl-5-hydroxytryptamine, or a combination thereof.

[0163] A method for reducing and / or inhibiting nerve cell damage and / or death

[0164] The present invention also provides a method for reducing and / or inhibiting damage and / or death of nerve cells, comprising the steps of: (a) providing a nerve cell; and (b) incubating the nerve cell with the drug or pharmaceutical composition, thereby reducing and / or inhibiting damage and / or death of the nerve cell.

[0165] Preferably, the method is performed in vivo and / or in vitro; preferably, the method is performed in vitro.

[0166] Preferably, the neural cells may be differentiated neural cells, such as neurons; or undifferentiated neural cells, such as neural crest cells. Preferably, the neural cells include neurons and neural crest cells. Preferably, the neural cells include dopaminergic neurons; and the neural crest cells include neuroblastoma cells. Preferably, the dopaminergic neurons include tyrosine hydroxylase (TH)-positive dopaminergic neurons.

[0167] As used herein, the term "dopaminergic neuron" refers to a type of neuron that synthesizes, stores, and releases dopamine. These neurons are located in brain regions such as the substantia nigra and hypothalamus and are involved in biological activities such as motor control, emotional regulation, and cognitive function. In Parkinson's disease patients, motor dysfunction occurs due to the degeneration of dopaminergic neurons in the substantia nigra. In this study, the number of TH-positive dopaminergic neurons decreased in an animal model of Parkinson's disease induced by MPTP.

[0168] As used herein, the term "neuroblastoma cells" refers to tumor cells derived from embryonic neural crest cells, which have cell heterogeneity and stem cell properties; have certain neuroendocrine functions and can secrete neurotransmitters. + In the induced Parkinson's disease cell model, the activity of neuroblastoma cells decreased.

[0169] Preferably, the neural cells are derived from humans or non-human mammals. The non-human mammals include rodents (such as mice and rats) and non-human primates (such as monkeys). Preferably, the neural cells are derived from humans or mice.

[0170] Preferably, the drug or pharmaceutical composition is selected from the group consisting of yamachulide K, N-p-coumaroyl-5-hydroxytryptamine, (Z)-bisabosterone, or a combination thereof. Preferably, the drug or pharmaceutical composition is yamachulide K. Preferably, the drug or pharmaceutical composition is N-p-coumaroyl-5-hydroxytryptamine. Preferably, the drug or pharmaceutical composition is (Z)-bisabosterone.

[0171] Preferably, the concentration of the drug or pharmaceutical composition is ≥5 μM.

[0172] Preferably, the reduction and / or inhibition of nerve cell damage and / or death comprises increasing nerve cell activity and / or increasing nerve cell proliferation capacity. Preferably, the improvement of nerve cell proliferation capacity comprises increasing the number of nerve cells and increasing the proportion of nerve cells. Preferably, the reduction and / or inhibition of nerve cell damage and / or death comprises increasing the activity and / or number of nerve cells.

[0173] Preferably, the drug or pharmaceutical composition can increase the activity of nerve cells by more than 1 times, such as ≥110%, ≥120%, ≥130%, ≥140%, compared to the activity of nerve cells observed in the absence of the drug or pharmaceutical composition.

[0174] Preferably, the drug or pharmaceutical composition can increase the number of nerve cells by more than 1 times, such as ≥110%, ≥120%, ≥130%, ≥140%, compared to the number of nerve cells observed in the absence of the drug or pharmaceutical composition.

[0175] Preferably, compared with the activity of nerve cells observed in the absence of or without the administration of the Nomadolactone K, the Nomadolactone K can increase the activity of nerve cells by more than 1 times, such as ≥140%.

[0176] Preferably, the N-p-coumaroyl-5-hydroxytryptamine can increase the activity of nerve cells by more than 1 times, such as ≥110%, compared to the activity of nerve cells observed in the absence of / without the administration of the N-p-coumaroyl-5-hydroxytryptamine.

[0177] Preferably, the (Z)-bisabolsterone can increase the activity of nerve cells by more than 1 times, such as ≥110%, compared to the activity of nerve cells observed in the absence of (Z)-bisabolsterone.

[0178] Preferably, the N-p-coumaroyl-5-hydroxytryptamine can increase the number of nerve cells by more than 1 times, such as ≥130%, compared to the number of nerve cells observed in the absence of / without the administration of the N-p-coumaroyl-5-hydroxytryptamine.

[0179] The main advantages of the present invention include:

[0180] (1) The present invention used the CCK-8 method to preliminarily screen and obtain 13 traditional Chinese medicine monomers with significant neuroprotective activity. Among them, two traditional Chinese medicine monomers that have not been reported in PD were discovered for the first time, namely, yamachulide K and N-p-coumaryl-5-hydroxytryptamine.

[0181] (2) The present invention uses prediction software to predict the properties of the screened Chinese medicine monomers, and for the first time discovered that yemazhui lactone K can pass through the blood-brain barrier, and has application prospects in the field of PD treatment.

[0182] (3) The present invention further verifies through animal experiments that N-p-coumaryl-5-hydroxytryptamine can significantly increase the survival rate of neurons in PD animal models.

[0183] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0184] Materials and methods

[0185] 1. Source of Chinese medicine monomers:

[0186] The monomer library of Chinese medicine for promoting blood circulation and removing blood stasis (Cat: HY-L193) was purchased from MCE Company.

[0187] 2. Construction of PD cell model:

[0188] 2mM and 3mM 1-methyl-4-phenylpyridinium ion (MPP + , Cat: D048) were used to induce human neuroblastoma SH-SY5Y cells for 24 h to construct a PD cell model.

[0189] 3. Set up the experimental group:

[0190] (1) Normal control group (Control group), (2) Model group (MPP + Group): Add 2mM or 3mM MPP + , (3) Drug treatment group: adding different Chinese medicine monomers.

[0191] 4. Preliminary screening of MPP +Candidate Chinese herbal monomers with protective effects on induced nerve damage:

[0192] (1) SH-SY5Y cells were seeded into 96-well plates and incubated in a 37°C, 5% CO2 incubator for 24 hours.

[0193] (2) The drug-treated group was treated with 2 mM or 3 mM MPP. + and 10 μM of different Chinese medicine monomers, and treated together for 24 hours.

[0194] (3) Finally, 10 μL of CCK-8 solution was added to each well and incubated for another 2 hours. After the incubation, the absorbance of each well was accurately measured at a wavelength of 450 nm using a microplate reader, and the cell viability was calculated based on the measured absorbance.

[0195] 5. Verify the effects of different administration methods and concentrations of the above-mentioned Chinese medicine monomers on the PD cell model:

[0196] (1) SH-SY5Y cells were seeded into 96-well plates and incubated in a 37°C, 5% CO2 incubator for 24 hours.

[0197] (2) Another administration method was used, that is, the model was established first and then the Chinese medicine monomer was administered for treatment: 3 mM MPP was added to both the model group and the drug treatment group. + The treatment lasted for 24 hours, and the old culture medium was discarded after the treatment. At this time, the drug treatment groups were treated with 5μM, 10μM, and 20μM Chinese medicine monomers, and the model group (i.e., MPP + After 24 hours of treatment, the old culture medium was discarded and normal culture medium was added.

[0198] (3) Finally, 10 μL of CCK-8 solution was added to each well and cultured for 1.5 hours. The absorbance of each well was measured at a wavelength of 450 nm using a microplate reader. The cell survival rate was calculated to screen out the MPP-sensitive cells. + Candidate Chinese herbal monomers with protective effects against induced nerve damage.

[0199] 6. Verification of the neuroprotective effects of yamachulide K and N-p-coumaryl-5-hydroxytryptamine in an MPTP-induced PD mouse model:

[0200] (1) Experimental groups: Wild-type eight-week-old male C57BL / 6J mice were randomly divided into the following groups: ① control group: injected with an equal amount of normal saline; ② MPTP group (subacute PD mouse model): intraperitoneal injection of 30 mg / kg of MPTP once a day for 5 consecutive days, for a total of 26 days; ③ yemachaylactone K group: based on the MPTP group, yemachaylactone K was intraperitoneally injected every day (high dose: 3 mg / kg, low dose: 1 mg / kg) for a total of 26 days; ④ N-p-coumaroyl-5-hydroxytryptamine group: based on the MPTP group, N-p-coumaroyl-5-hydroxytryptamine was intraperitoneally injected every day (high dose: 20 mg / kg, low dose: 10 mg / kg) for a total of 26 days.

[0201] (2) Behavioral testing:

[0202] (i) Rotarod fatigue test: The exercise capacity of mice was assessed using a rotarod fatigue test. The rotation speed of the rotarod was gradually increased from 0 to 30 rpm for 5 minutes. Each mouse was tested three times, with a 1-minute interval between each test. The time the mouse remained on the rotarod was recorded to assess its exercise endurance.

[0203] (ii) Pole climbing test: Mice were placed on a rough wooden ball, the lower end of which was attached to a rough, round wooden stick placed in a cage. The mouse was placed head-up on the top of the pole, and the time required for the mouse to turn downward (Tturn) and descend from the top to the bottom of the pole (Tfall) was recorded. Each mouse was tested three times, and the average of the three climbing times was used as a statistical indicator to assess the mouse's motor coordination ability.

[0204] (iii) Open field test: The mouse was placed in the center of a 16-grid box, and a video camera and timing device were activated. Video recording was stopped after 5 minutes of observation. The total distance the mouse moved within the box and the percentage of time spent in the center grid were analyzed to assess its anxiety level and ability to move autonomously.

[0205] Example 1: MPP + Screening of candidate Chinese herbal monomers with protective effects on induced nerve damage

[0206] This embodiment involves MPP + Candidate Chinese medicine monomers with protective effects in inducing nerve damage are screened. The specific steps are as follows:

[0207] First, SH-SY5Y cells were seeded into 96-well plates and then incubated in a 37°C, 5% CO2 incubator for 24 hours to ensure that the cells were fully attached to the wall. After the cells were fully attached, the experiment was divided into three groups: the experimental group was added with 2mM MPP +and 10μM different Chinese medicine monomers, and continued to treat for 24 hours; the normal control group (control group) was only added with culture medium; the model group (MPP + Group) Add 2 mM MPP + Next, the CCK-8 method was used to detect cell activity: 10 μL of CCK-8 solution was added to each well and cultured for 2 hours. After the culture was completed, the absorbance of each well was measured at a wavelength of 450 nm using an enzyme marker. The cell survival rate was calculated based on the measured absorbance, and the cells that may be active against MPP were preliminarily screened. + Candidate Chinese herbal monomers with protective effects against induced nerve damage.

[0208] To further verify the accuracy and reliability of the preliminary screening results, 3 mM MPP was used + The cells were synchronously treated with 10 μM of different Chinese medicine monomers, and the above series of experimental steps were repeated to ensure that the screened Chinese medicine monomers did have a protective effect on nerve damage.

[0209] The above method was used to preliminarily screen 165 Chinese medicine monomers ( Figure 1 ), identified 13 candidate ingredients with significant neuroprotective activity, including Dehydrocorydaline nitrate, 11-Keto-beta-boswellic acid, Loganin, Butein, Eupalinolide K, Curcumin, Iristectorigenin B, (Z)-Guggulsterone, Arachidonic acid, Evodiamine, Acacetin, N-(p-Coumaroyl)Serotonin, and Kushenol I. Among them, Eupalinolide K, (Z)-Guggulsterone, and N-(p-Coumaroyl)Serotonin significantly alleviated MPP. + Induced toxicity, and has not been seen or rarely reported in PD.

[0210] In addition, the results showed that even at a higher concentration (3mM) of MPP + Under induction conditions, the simultaneous administration of the Chinese medicine monomers (Z)-bisabolsterone, yamachulide K and N-p-coumaryl-5-hydroxytryptamine can still significantly reverse MPP. + Induced cell death ( Figure 2 ).

[0211] Example 2: The traditional Chinese medicine monomers yemazhui lactone K, N-p-coumaryl-5-hydroxytryptamine and (Z)-bisabolsterone have neuroprotective effects

[0212] This example involves further exploring the effects of different concentrations of Chinese medicine monomers on a PD cell model under another administration method.

[0213] First, 3 mM MPP + The cells were treated for 24 hours to simulate a nerve injury-inducing environment. Subsequently, the cells were treated with different concentrations of the traditional Chinese medicine monomers (Z)-bisabolsterone, yamachulide K, and N-p-coumaryl-5-hydroxytryptamine for 24 hours. Detailed procedures are described in Materials and Methods 5.

[0214] The experimental results showed that this administration method can still significantly improve the survival rate of PD cell models ( Figure 3 This discovery provides key experimental support for in-depth research on the neuroprotective mechanism of Chinese medicine monomers, indicating that Chinese medicine monomers (Z)-bisabolsterone, yamachulide K and N-p-coumaryl-5-hydroxytryptamine have a significant effect on MPP. + It induces positive intervention efficacy in neuronal damage and plays a protective role.

[0215] In order to evaluate the possibility of candidate monomers to play a role in the central nervous system, this application also uses three tools: pkCSM, SwissADME, and admetSAR3.0 for prediction.

[0216] The comprehensive prediction results show that the Chinese medicine monomers yemazhui lactone K and (Z)-bisabolsterone are likely to penetrate the blood-brain barrier, while the Chinese medicine monomer N-p-coumaryl-5-hydroxytryptamine may not penetrate the blood-brain barrier ( Figure 4-6 ).

[0217] Example 3: Yemazhui lactone K and N-p-coumaryl-5-hydroxytryptamine significantly alleviate MPTP-induced damage to tyrosine hydroxylase (TH)-positive dopaminergic neurons and promote neuronal survival

[0218] This example involves verifying the neuroprotective effects of two traditional Chinese medicine monomers, yemazhuilactone K and N-p-coumaryl-5-hydroxytryptamine, in an animal PD model.

[0219] First, a PD animal model was established using MPTP, and animals treated with normal saline were used as the control group. Subsequently, the PD model group was given intervention therapy with yemachalide K and N-p-coumaryl-5-hydroxytryptamine. For specific procedures, see Materials and Methods 6.

[0220] The results showed that the administration of N-p-coumaryl-5-hydroxytryptamine significantly increased the number of TH-positive dopaminergic neurons in the PD animal model. This finding indicates that N-p-coumaryl-5-hydroxytryptamine has the effect of promoting neuronal survival. In addition, N-p-coumaryl-5-hydroxytryptamine can also shorten the T-turn residence time in the climbing pole test of PD mice, indicating that this compound can also improve the motor coordination ability of PD mice ( Figure 7 ).

[0221] In summary, these research results demonstrate that N-p-coumaryl-5-hydroxytryptamine can effectively alleviate PD motor deficits, increase the survival rate of dopamine neurons, and play a neuroprotective role in PD.

[0222] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A drug or pharmaceutical composition, characterized in that The drug or pharmaceutical composition comprises: (1) Eupalinolide K; (2) N-(p-Coumaroyl)Serotonin; (3) (Z)-Guggulsterone; or a combination thereof.

2. The drug or pharmaceutical composition according to claim 1, wherein The drug or pharmaceutical composition further comprises: (4)Dehydrocorydaline nitrate; (5)11-Keto-beta-boswellic acid; (6) Loganin; (7) Butein; (8) Curcumin; (9)Iristectorigenin B; (10)Arachidonic acid; (11) Evodiamine; (12)Acacetin; (13) Kushenol I; or a combination thereof.

3. The drug or pharmaceutical composition according to claim 1, wherein The drug or pharmaceutical composition comprises: (a) a therapeutically and / or prophylactically effective amount of yamadrolactone K, N-p-coumaryl-5-hydroxytryptamine, (Z)-bisabolsterone, or a combination thereof; (b) a pharmaceutically acceptable carrier, diluent or excipient.

4. The drug or pharmaceutical composition according to claim 3, wherein The drug or pharmaceutical composition further comprises: (c) a therapeutically and / or prophylactically effective amount of Dehydrocorydaline nitrate, 11-Keto-beta-boswellicacid, Loganin, Butein, Curcumin, Iristectorigenin B, Arachidonic acid, Evodiamine, Acacetin, Kushenol I, or a combination thereof.

5. Use of the medicament or pharmaceutical composition according to any one of claims 1 to 4, characterized in that: Used for preparing drugs for treating and / or preventing neurodegenerative diseases.

6. The use according to claim 5, characterized in that The neurodegenerative diseases include Parkinson's disease.

7. A medicine box, characterized in that: include: (A) a first kit, and the drug or pharmaceutical composition according to any one of claims 1 to 4 located in the first kit; (B) a second drug kit, and other drugs for treating neurodegenerative diseases in the second drug kit.

8. A method for reducing and / or inhibiting nerve cell damage and / or death, characterized in that: Including steps: (a) providing a nerve cell; (b) incubating the nerve cells with the drug or pharmaceutical composition according to any one of claims 1 to 4, thereby reducing and / or inhibiting damage and / or death of the nerve cells.

9. The method according to claim 8, wherein The drug or pharmaceutical composition is selected from the following group: yamachulide K, N-p-coumaryl-5-hydroxytryptamine; the concentration of the drug or pharmaceutical composition is ≥5 μM.

10. The method according to claim 8, wherein The reducing and / or inhibiting the damage and / or death of the nerve cells includes: increasing the activity and / or number of the nerve cells; incubating the nerve cells with the yamachulide K, so that the activity and / or number of the nerve cells are significantly increased.

Images