Pharmaceutical combination for preventing or treating epileptic seizure and use thereof

Abstract

To provide a herbal medicine composition that serves as a new option for preventing or treating epileptic seizure, and is combined with other anti-epileptic drugs to reduce side effects of the anti-epileptic drugs.SOLUTION: The present disclosure provides a herbal medicine composition for preventing or treating epileptic seizure and use in the preparation of a medication. The herbal medicine composition has the characteristics of high efficiency, safety, and convenience, can effectively prevent or treat epileptic seizure, and may be further combined with other anti-epileptic drugs to enhance the anti-epileptic effects and reduce side effects of the anti-epileptic drugs. The present disclosure also provides a method of preventing or treating epileptic seizure by using the herbal medicine composition or a pharmaceutical combination thereof.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a combination of crude drugs for preventing or treating epileptic seizures and its uses, and also relates to a pharmaceutical combination comprising a combination of crude drugs and at least one antiepileptic drug and its uses.

Background Art

[0002] The description of the background art contains information helpful for understanding the present invention, and all information provided in this specification should not be regarded as the background art or the technology related to the present invention. Also, any publications cited explicitly or implicitly should not be regarded as the background art.

[0003] Epilepsy is a common neurological disorder that plagues approximately 70 million people worldwide. The characteristics of this disease are recurrent, spontaneous, and unpredictable seizures caused by abnormal discharges of brain nerve cells, mainly occurring in the cerebral cortex and hippocampus. The causes of epilepsy are related to congenital or acquired brain lesions, such as brain damage, infection, stroke, brain tumor, etc., but the seizures of 70% of patients are of unknown origin. Although the cause of most epilepsy is unknown, it is generally considered that the balance between the brain inhibitory nervous system (i.e., GABA system) and the excitatory nervous system (i.e., glutamate system) is disrupted, causing excessive excitation of brain nerve cells.

[0004] Currently, the mechanisms of action of anti-epileptic drugs that suppress epileptic seizures mainly include drugs that directly suppress the excitability of brain nerve cells by blocking sodium channels (e.g., phenytoin, carbamazepine), drugs that suppress glutamate (e.g., lamotrigine, felbamate), or drugs that increase GABA (e.g., benzodiazepines, tiagabine, vigabatrin), etc., to regulate the stability of brain nerve cells. However, there are currently nearly 30 types of anti-epileptic drugs, but about one-third of epilepsy patients have drug resistance to the therapeutic drugs, and the side effects of the drugs also affect the patient's adherence to the current therapeutic drugs used.

[0005] Currently, it is known that commonly used anti-epileptic drugs have many side effects. For example, in the package insert of carbamazepine, side effects such as drowsiness, loss of appetite, diarrhea, dizziness, heaviness in the head, ataxia (coordination movement disorder), leukopenia, aplastic anemia, liver dysfunction, arrhythmia, and rash are described. In the package insert of phenytoin, side effects such as nausea, vomiting, heartburn, loss of appetite, diplopia, nystagmus, ataxia (coordination movement disorder), gingival hyperplasia, hirsutism, facial roughness, involuntary movement, lymph node swelling, malformation, and rash are described. The usual treatment methods after side effects occur are to change the drug, reduce the dosage, or add other drugs to suppress the side effects. However, reducing the dosage makes epilepsy likely to recur, which is painful for the patient both physically and mentally.

[0006] Therefore, it is necessary to search for and develop new drugs that are more effective and safe in the treatment of epilepsy. Medicinal plants have attracted attention because they are widely used in traditional and folk medicine for the treatment and prevention of different nervous system diseases (including epilepsy). In recent years, many plant-derived bioactive molecules have been proven to have an anti-epileptic effect by affecting ion channels, GABA, or glutamate. For example, flavonoids, alkaloids, terpenoids, cannabidiol, etc., medicinal plants have become the research targets of anti-epileptic drugs.

[0007] Taiwan Qingguan No. 1 (NRICM101) is a herbal medicine developed by the National Research Institute of Chinese Medicine in 2020 based on the prescription of "Jingfang Baidu Powder" in the "Shengbai Zhongmiaofang" of the Ming Dynasty and is used for the treatment of the novel coronavirus (COVID-19). It has antiviral, immune regulation and anti-inflammatory effects. Taiwan Qingguan No. 1 is prepared by mixing 10 herbal medicines including Scutellaria Root (Scutellaria baicalensis), Heartleaf Houttuymia (Houttuynia cordata), Indigowoad Root (Isatis indigotica), Mongolian Snakegourd Fruit (Trichosanthes kirilowii), Mulberry Leaf (Morus alba), Magnolia Bark (Magnolia officinalis), Peppermint Herb (Mentha haplocalyx), Fineleaf Nepeta (Nepeta tenuifolia), Saposhnikovia Root (Saposhnikovia divaricate) and Baked Liquorice Root (Glycyrrhiza glabra). Furthermore, various bioactive components have also been discovered in Taiwan Qingguan No. 1. For example, baicalein, epigoitrin, liquiritin, quercetin-3-galactoside, quercetin-3-isorhamnetin, scutellarin, rutin, oroxindin, chlorogenic acid, etc. These components may be the cause of the pharmacological properties of Taiwan Qingguan No. 1. However, there is still no data exploring the anti-epileptic effect of Taiwan Qingguan No. 1. Such an anti-epileptic effect may provide a new option for preventing or treating epileptic seizures, or may reduce the side effects of conventional anti-epileptic drugs by combining with other anti-epileptic drugs.

Summary of the Invention

Problems to be Solved by the Invention

[0008] The problem to be solved by the present invention is to provide a crude drug composition that serves as a new option for preventing or treating epileptic seizures and reduces the side effects of anti-epileptic drugs when combined with other anti-epileptic drugs.

Means for Solving the Problem

[0009] The present invention provides a crude drug composition having the use for preventing or treating epileptic seizures, and the crude drug composition contains saffron, houttuynia, schizonepeta, carthamin, perilla, cinnamon, peppermint, schizandra, bupleurum, and licorice.

[0010] In one embodiment, epilepsy is caused by damage to brain nerve cells. In another embodiment, epilepsy is caused by the activation of glial cells. In another embodiment, epilepsy is caused by an inflammatory reaction in the brain by inflammatory molecules. In another embodiment, the inflammatory molecules are selected from the group consisting of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), high mobility group box 1 (HMGB1), interleukin-1 receptor 1 (IL-1R1), and Toll-like receptor-4 (TLR-4).

[0011] In one embodiment, the crude drug composition of the present invention is further combined with at least one anti-epileptic drug to be manufactured into a drug for preventing or treating epileptic seizures. In another embodiment, the at least one anti-epileptic drug is selected from Western medicines combined with phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepine, tiagabine, and vigabatrin.

[0012] The present invention provides a combination of pharmaceuticals for preventing or treating epileptic seizures, comprising a crude drug composition, at least one antiepileptic drug, and a pharmaceutically acceptable carrier and / or stabilizer. The crude drug composition comprises saffron, houttuynia, schizonepeta, carthamin, perilla, cinnamon, peppermint, schizandra, bupleurum, and licorice. Also, when treating epilepsy, the dosage of the antiepileptic drug is less than the dosage when the antiepileptic drug is used alone. In one embodiment, the at least one antiepileptic drug is selected from the group consisting of a combination of phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepine, tiagabine, and vigabatrin. In one embodiment, the crude drug composition and the at least one antiepileptic drug are administered to a patient simultaneously, sequentially, or at intervals.

[0013] Each object, feature, aspect, and excellent point of the present invention will become clearer from the following detailed description and drawings in the embodiments.

Effects of the Invention

[0014] The crude drug composition of the present invention serves as a drug for preventing or treating epileptic seizures, and by combining with other antiepileptic drugs, it reduces the side effects of the antiepileptic drugs.

Brief Description of the Drawings

[0015]

Figure 1

Figure 2

Figure 3A

Figure 3B

DETAILED DESCRIPTION OF THE INVENTION

[0016] The features and advantages of the present invention will become more apparent from the following description of the drawings and better examples, where similar reference terms in the text refer to the same parts or components.

[0017] The following description contains information useful for understanding the present invention, and all information provided herein should not be considered as related to the prior art or the scope of the claims of the present invention, and any publications cited explicitly or implicitly should not be considered as prior art.

[0018] The present invention is not limited to the specific examples of materials, structures, processes, methods, or structures mentioned in this specification. Therefore, although several options are given in this specification, all those similar or having the same effect can be applied to the implementation methods or examples of the present invention, and the best materials and methods are described in this specification.

[0019] The following detailed description related to the accompanying drawings is one of the exemplary embodiments of the present invention and is not intended to represent the only exemplary embodiment for implementing the present invention. The technical term "exemplary" used in this specification means "an example, instance, or case", and it goes without saying that it is not better or superior to other possible exemplary embodiments. The detailed description includes specific parts and is provided for understanding the embodiments in this specification. For those skilled in the art, the embodiments in this specification can be implemented without using these specific parts.

[0020] As used in this specification, each of the following technical terms includes the meaning described in that paragraph. The technical terms used in the present invention are for the purpose of explaining specific embodiments of the present invention and are not intended to be restrictive.

[0021] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In general, the nomenclature used herein is well known and commonly employed in the laboratory procedures of animal pharmacology, pharmaceutical sciences, separation science, and organic chemistry. In the methods described herein, operations can be performed in any order, unless the order of time and operations is clearly described. Further, unless explicitly stated to the contrary in the vocabulary description of the claims, the operations are meant to be performed simultaneously. For example, the operation of performing X and the operation of performing Y can be carried out simultaneously in a single operation, and the resulting method is also within the scope of the claimed method.

[0022] The following non-limiting initials are used herein: KA, kainic acid; IL-1β, interleukin-1β; IL-6, interleukin-6; TNF-α, tumor necrosis factor-α; HMGB1, high mobility group Box 1; IL-1R1, interleukin-1 receptor 1; CBZ, carbamazepine; CABA, gamma-aminobutyric acid; TLR-4, Toll-like receptor-4; HPLC, high-performance liquid chromatography.

[0023] Unless clearly indicated to the contrary in context, the terms "a", "an", and "the" as used in this specification and the claims include plural referents. For example, "a component" means one component or more than one component.

[0024] As used herein, one of ordinary skill in the art should be able to understand the technical term "about" and it can vary to some extent depending on the context. When used herein, in the case of measurable values such as quantity, length of time, etc., "about" means including a variation of ±20%, ±10%, ±5%, ±1% or ±0.1% of the specific value. These variations are suitable for the disclosed methods. For this reason, in some embodiments, the numerical parameters in this specification and the claims are approximate values and vary in order to obtain the expected characteristics by a specific implementation method. In some embodiments, the numerical parameters are interpreted by applying general numerical rounding techniques according to the significant digits described. The numerical values in some embodiments of the present invention may have errors, and the cause is due to the standard deviation in the test measurements respectively.

[0025] In one aspect, the technical term "co - administration" related to the subject means that while administering the herbal medicine composition of the present invention to the subject, other agents that treat or prevent the diseases considered herein are also administered. In some embodiments, the herbal medicine composition and other agents to be co - administered are administered separately or in any kind of combination as part of some single - treatment methods.

[0026] As used herein, "disease" is a kind of the health state of the subject. The subject cannot maintain the stable state in the body, and if the disease is not improved, the health of the subject will continue to deteriorate.

[0027] As used herein, the "medical condition" of the subject is a kind of health state. The health state of the subject is not better than when there is no medical condition, but the subject can maintain the stable state in the body. Even if the medical condition is not treated, the health state of the subject does not necessarily deteriorate.

[0028] As used herein, the technical term "pharmaceutically acceptable" means a substance that does not eliminate the biological activity or properties of the useful compounds in the present invention, such as a vehicle or a diluent. Further, the substance is non - toxic, that is, administering the substance to the subject does not cause an unexpected biological reaction, or all components contained in the composition do not interact in a harmful way.

[0029] As used herein, the technical terms of a compound, "effective dose in a pharmaceutical", "therapeutically effective dose" or "effective dose" refer to the amount of the compound that brings a beneficial effect to the subject administered with the compound.

[0030] As used herein, the technical terms "subject", "individual" and "patient" are interchangeable with each other and include humans or non-human mammals. Non-human mammals include, for example, livestock and pets, such as sheep, pigs, canines, felines and murine mammals. In some embodiments, the subject is human.

[0031] As used herein, the technical term "herbal composition" refers to a combination of at least two or more herbs having therapeutic effects, and each herb is directly mixed after being ground, or each herb is mixed before or after conventional pharmaceutical processing, extraction, concentration and / or other preparation methods.

[0032] As used herein, the technical terms "pharmaceutical combination" or "pharmaceutical composition" refer to a combination of at least two or more pharmaceuticals, which are herbs and / or Western medicines.

[0033] As used herein, the technical terms "prevention" or "prevention" mean that, at the time when a pharmaceutical or compound is administered, the subject is before showing a disease or medical condition, and the occurrence of these symptoms is avoided or delayed.

[0034] As used herein, the technical term "treatment" means reducing the frequency or progression of a disease or medical condition experienced by a subject by administering a pharmaceutical or compound to the subject.

[0035] As used herein, "therapeutic effectiveness" is a beneficial therapeutic response, for example, relieving or improving one or some symptoms of a disease; reducing the degree of a disease; delaying or alleviating the progression of a medical condition; improving, reducing or stabilizing a medical condition; and other beneficial results.

[0036] Throughout the disclosed content, each aspect of the present invention is presented in a range format. The range format description is for convenience and brevity only and is not a limitation on the scope of the present invention. For this reason, the range description is specifically regarded as all possible sub-ranges and the individual numerical values within that range. For example, the description of the range from 1 to 6 is regarded as specifying sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and also the individual and fractional numbers within that range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. All apply regardless of the range.

[0037] The crude drug composition of the present invention is based on Qingguan No. 1 and contains 10 crude drugs such as Scutellaria baicalensis, Houttuynia cordata, Isatis indigotica, Trichosanthes kirilowii, Morus alba, Magnolia officinalis, Mentha haplocalyx, Nepeta tenuifolia, Saposhnikovia divaricate, and Glycyrrhiza glabra.

[0038] In one embodiment, the crude drug composition of the present invention comprises 3.75 to 1.25 parts by weight of saffron, 3.75 to 1.25 parts by weight of houttuynia cordata, 3.75 to 1.25 parts by weight of schizonepeta tenuifolia, 3.75 to 1.25 parts by weight of calonyction aculeatum, 2.25 to 0.75 parts by weight of perilla frutescens, 2.25 to 0.75 parts by weight of cinnamon, 2.25 to 0.75 parts by weight of peppermint, 2.25 to 0.75 parts by weight of gentiana scabra, 1.5 to 0.5 parts by weight of bupleurum chinense, and 1.5 to 0.5 parts by weight of licorice. In another embodiment, a preferred combination of the crude drug composition of the present invention is 2.75 to 2.25 parts by weight of saffron, 2.75 to 2.25 parts by weight of houttuynia cordata, 2.75 to 2.25 parts by weight of schizonepeta tenuifolia, 2.75 to 2.25 parts by weight of calonyction aculeatum, 1.65 to 1.35 parts by weight of perilla frutescens, 1.65 to 1.35 parts by weight of cinnamon, 1.65 to 1.35 parts by weight of peppermint, 1.65 to 1.35 parts by weight of gentiana scabra, 1.1 to 0.9 parts by weight of bupleurum chinense, and 1.1 to 0.9 parts by weight of licorice. In another embodiment, the crude drug composition of the present invention preferably contains 2.5 parts by weight of saffron, 2.5 parts by weight of houttuynia cordata, 2.5 parts by weight of schizonepeta tenuifolia, 2.5 parts by weight of calonyction aculeatum, 1.5 parts by weight of perilla frutescens, 1.5 parts by weight of cinnamon, 1.5 parts by weight of peppermint, 1.5 parts by weight of gentiana scabra, 1 part by weight of bupleurum chinense, and 1 part by weight of licorice.

[0039] The crude drugs contained in the crude drug composition of the present invention are manufactured into liquid preparations through processes such as washing, drying, grinding, and processing. In one embodiment, the liquid preparation is further manufactured into a solid preparation through processes such as extraction, concentration, addition of excipients, and spray granulation. The above manufacturing processes can be increased or decreased according to the dosage forms manufactured by conventional crude drug pharmaceutical technologies.

[0040] In one aspect, the crude drug composition of the present invention can be used in a combination of one or more kinds of pharmaceuticals for preventing or treating epilepsy, or manufactured into a combination of pharmaceuticals. These drugs include known compounds (such as purchasable compounds) for treating, preventing, or alleviating epilepsy symptoms.

[0041] In one embodiment, the present invention provides a method for treating or preventing epilepsy in a subject. This method includes administering the herbal medicine composition of the present invention to the subject in a therapeutically effective amount. In another embodiment, the subject is further co-administered with at least one drug effective for treating or preventing epilepsy. In another embodiment, at least one drug includes, for example, phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepine, tiagabine, vigabatrin, etc. The dosage of Western medicine can be formulated by a doctor according to the recommended dosage of each drug.

[0042] In one embodiment, the subject with epilepsy is a person who has been newly diagnosed with epilepsy or a person who is taking at least one anti-epileptic drug. In one embodiment, when a combination of the herbal medicine composition of the present invention and at least one anti-epileptic drug is given to a subject newly diagnosed with epilepsy and at least one anti-epileptic drug is administered in combination for prevention or treatment, the dosage is less than the dosage when the anti-epileptic drug is administered alone. In another embodiment, the herbal medicine composition of the present invention is given to a subject who is taking at least one anti-epileptic drug, and further the dosage of the anti-epileptic drug can be reduced.

[0043] The treatment method affects an effective amount of the combination. The therapeutic formulation can be administered to the patient before or after the onset of the disease or symptom. Further, it can be administered daily, or in divided doses at several times and in alternating shifted doses in sequence, or the doses can be administered continuously. In addition, according to the urgency of the treatment or prevention situation, the dosage of the proportional therapeutic formulation can be increased or decreased.

[0044] By administering the herbal medicine composition of the present invention to a patient in an already known order, dosage, and period, epilepsy can be effectively treated or prevented. The effective amount of the herbal medicine composition required for the effect of treating and / or preventing epilepsy varies depending on many factors, such as the activity of the specific herbal medicine composition used; the administration time; the excretion rate of the extract; the duration of treatment; other drugs, compounds, or substances used in combination with the extract; the state of the disease or medical condition before the patient receives treatment, the patient's age, gender, weight, symptoms, general health status, and previous medical history; similar factors well-known in the medical field, etc. The dosage, etc. is adjusted to provide an excellent therapeutic response. For example, a dosage divided into several times a day is administered, or the dosage is decreased in proportion to the urgency of the treatment situation.

[0045] The description of the herbal medicine composition of the present invention is mainly for administration to humans in a method suitable for medical ethics. For those skilled in the art, it should be understandable that the herbal medicine composition is usually suitable for various animals. In order to be able to administer to various animals, it is well-known that the recombination of the herbal medicine composition of the present invention can be carried out, and veterinary pharmacists familiar with the technology can design and implement such recombination by general experiments (if necessary). The subjects to which the herbal medicine composition of the present invention can be administered include humans and other primates, non-mammals, commercially valuable mammals, such as cows, pigs, horses, sheep, cats, and dogs, etc., without limitation.

[0046] In some embodiments, the herbal medicine composition of the present invention is administered to a patient at a dosage of 1 to 5 times a day or even more. In other embodiments, the herbal medicine composition of the present invention is administered to a patient at a dosage of once a day, twice a day, three times a day, four times a day, once every two days, once every three days to one week, or once every two weeks. It is easy for those skilled in the art to understand that the administration frequency of each combination composition of the present invention varies depending on the tester, and the administration frequency is affected by many factors, including age, the disease or medical condition to be treated, gender, overall health status, and other factors, without limitation. Therefore, the present invention should not be construed as being limited to any specific dosage plan, and the attending physician should consider all factors related to the patient and confirm the exact dosage and composition to be administered to the patient.

[0047] In one embodiment, when the crude drug composition of the present invention is administered in combination with at least one anti-epileptic drug, the crude drug composition and at least one anti-epileptic drug can be simultaneously administered to the subject. In another embodiment, the crude drug composition and at least one anti-epileptic drug can be administered to the subject in sequence, before and after, or with an interval therebetween.

[0048] Administration method: Oral administration; The dosage form of the crude drug composition of the present invention for oral administration is solid or liquid. Suitable solid dosage forms include, for example, powders, fine granules, granules, pills, tablets, capsules, lozenges, etc. Suitable liquid preparations include solutions, syrup preparations, suspensions, dispersions, concentrated solutions, etc. These dosage forms are manufactured by selecting and adding pharmaceutically acceptable additives according to conventional pharmaceutical methods.

[0049] Those skilled in the art should be able to understand that by conducting experiments within the normal range, equivalent items such as the specific order, implementation method, scope of claims, and examples described in the present invention can be confirmed. These equivalent items are recognized within the scope of the present invention and are further included in the scope of the claims. For example, modifications of reaction conditions such as reaction time, reaction state / volume, and experimental reagents (such as solvents, catalysts), pressure, atmospheric pressure conditions, and reducing / oxidizing agents, as well as alternatives recognized in the technical field and experiments within the normal range, are all within the scope of the present invention.

[0050] Regarding all numerical values and ranges mentioned in this specification, the description of the range is for convenience and brevity and should not be construed as a limitation on the scope of the present invention. Therefore, all numerical values and ranges included in the numerical values and ranges are included within the scope of the present invention. Also, the upper or lower limits of all numerical values and numerical ranges falling within these ranges are also within the contemplation of the present invention. The description of the range specifically discloses all possible sub-ranges and individual numerical values within the range, and in appropriate cases, some integers of the numerical values are also included within the range. For example, the description of the range from 1 to 6 is considered to specifically disclose sub-ranges, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and individual numbers within the range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6, etc., are all applicable regardless of the range.

[0051] The following examples further illustrate various aspects of the present invention. However, they are not a limitation on the teachings or disclosure of the present invention described herein.

[0052] The present invention will be described with reference to the following examples. These examples are for illustrative purposes only, and the present invention is not limited by these examples. All variations that include the teachings provided herein will be apparent.

[0053] The crude drug compositions exemplified in the present invention are manufactured and / or tested by the following steps.

[0054] The crude drug composition used in one example uses commercially available drugs (Shuntenro, Taiwan Qingguan No. 1). One dose contains 2.5 parts by weight of safflower, 2.5 parts by weight of houttuynia, 2.5 parts by weight of schizonepeta, 2.5 parts by weight of carthamus, 1.5 parts by weight of perilla, 1.5 parts by weight of cinnamon, 1.5 parts by weight of peppermint, 1.5 parts by weight of schizonepeta tenuifolia, 1 part by weight of bupleurum, and 1 part by weight of licorice.

[0055] Male Sprague-Dawley rats (SD, 150 - 200 g) are purchased from BioLASCO TAIWAN Co., Ltd and housed in the Experimental Animal Center of Fu Jen Catholic University. All animal experiments have obtained permission from the Experimental Animal Management Committee of Fu Jen Catholic University. In accordance with the 3R rules, all experiments are conducted with the minimum number of animals and with the least possible animal suffering while producing reliable results.

[0056] [Component Analysis of Example 1 - NRICM101] Regarding the chemical characteristics of plants in Taiwan Qingguan No. 1 (NRICM101), some standard compounds were selected from the literature, and some active compounds were further identified from the methanol extract. The structures of these compounds were confirmed by 3D fingerprint analysis.

[0057] For high-performance liquid chromatography (HPLC) analysis, 20 mL of 70% methanol was sonicated at 25 °C for 20 minutes to extract 0.5 g of NRICM101. Then, the NRICM101 sample was filtered through a 0.45 μm syringe filter.

[0058] The Waters HPLC system (Milford, Massachusetts, USA) is composed of a Waters 600 pump system, a Waters 2996 photodiode array detector, a Waters 717 autosampler, and a Sugai U-620 column heater (Wakayama, Japan). The stationary phase is a combination of a Cosmosil 5C18-MS-II packed column (5 μm, 4.6 mm × 250 mm, Nacalai tesque, Japan) and a Lichrospher RP-18 high-performance liquid chromatography column (5 μm, 4.0 mm × 10 mm, Merck, Germany). Gradient elution is combined from eluents A, B, and C (A: H2O / KH2PO4 / 10% H3PO4 = 1000 mL / 2.72 g / 1 mL; B: acetonitrile; C: H2O), and gradient elution is performed as follows: from 0 to 30 minutes, 90% - 75% A and 10% - 25% B; from 30 to 40 minutes, 75% - 65% A and 25% - 35% B; from 40 to 55 minutes, 65% - 0% A, 35% - 75% B, and 0% - 25% C; from 55 to 60 minutes, 75% - 10% B and 25% - 90% C; from 60 to 65 minutes, 0% - 90% A, 10% - 10% B, and 90% - 0% C. Gradient elution is used for 3D fingerprint analysis, and the peaks of liquiritin (25.94 minutes, 280 nm), rosmarinic acid (29.44 minutes, 320 nm), baicalin (34.61 minutes, 280 nm), oroxylin A-7-O-glucuronide (40.39 minutes, 280 nm), (wogonin-7-O-glucuronide) (42.53 minutes, 280 nm), glycyrrhizic acid (51.37 minutes, 250 nm), and magnolol (61.79 minutes, 290 nm) are detected. The flow rate is maintained at 1 mL / min, and the column temperature is maintained at 35 °C.

[0059] Figure 1 shows the chromatography of NRICM101. Among the freeze-dried extracts of NRICM101, the contents of baicalin, oroxylin A-7-O-glucuronide, wogonin-7-O-glucuronide, glycyrrhizic acid, liquiritin, magnolol, and rosmarinic acid are 18.34, 1.33, 2.67, 2.42, 1.70, 1.07, and 0.91 mg / g, respectively.

[0060] [Model of animal epilepsy induced by Example 2-KA] The anti-epileptic effect of the crude drug composition of the present invention is evaluated using an animal epilepsy induction model by kainic acid (KA). KA is an analogue of glutamic acid. When KA is injected into rats, the release of glutamic acid is increased, and the glutamic acid receptor is overly activated, thereby causing progressive limbic seizures. This causes an increase in calcium ions in nerve cells, oxidative stress, and mitochondrial function defects, leading to the death of nerve cells in many regions of the brain, especially in the cerebral cortex and hippocampal area. Furthermore, KA is related to the induction of an inflammatory response consisting of nerve cell death, activation of glial cells, and inflammatory-related molecules (e.g., interleukin 1β (IL-1β), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), high-mobility group protein (HMGB1), interleukin 1 receptor 1 (IL-1R1), toll-like receptor 4 (TLR-4)). Since these pathological changes induced by KA are similar to human epileptic behavior, the model of animal epilepsy induced by KA is a new compound widely used in screening for anti-epileptic activity.

[0061] After injecting KA into the abdominal cavity of rats, the epileptic behavior within 4 hours is analyzed. The evaluation method of seizure severity is based on the Racine scale (1972) and is shown in Table 1.

[0062]

Table 1

[0063] [Evaluation of the Effect of the Herbal Medicine Composition on Animal Seizure Behavior Induced by Example 3-KA] To evaluate the effect of the herbal medicine composition on animal seizure behavior induced by KA, physiological saline, the herbal medicine composition, and CBZ were administered daily at a fixed time by pipette, and KA (15 mg / kg) was intraperitoneally injected 7 days later. The seizure behavior was analyzed within 4 hours after the intraperitoneal injection of KA into the rats, and the seizure behavior was evaluated according to the above Racine scale (1972).

[0064] Different test substances are administered to Sprague-Dawley rats and divided into the following groups. 1. Negative control group: Physiological saline. 2. KA group: The test substance is not administered before injecting kainic acid (KA). 3. Low-dose herbal medicine composition group: 100 mg / kg of the herbal medicine composition. 4. Medium-dose herbal medicine composition group: 200 mg / kg of the herbal medicine composition. 5. High-dose herbal medicine composition group: 300 mg / kg of the herbal medicine composition. 6. Antiepileptic drug group: 100 mg / kg of carbamazepine (CBZ). 7. Antiepileptic drug composition group: Contains 100 mg / kg of the herbal medicine composition and 50 mg / kg of carbamazepine (CBZ).

[0065] Rats in each group are given the test substance daily at a fixed time by pipette, physiological saline is given to the control group, the herbal medicine composition is given to the herbal medicine composition group, carbamazepine is given to the antiepileptic drug group, and the herbal medicine composition and carbamazepine are given to the antiepileptic drug composition group. The herbal medicine composition and carbamazepine are mixed with physiological saline in powder form and then given to the rats. 7 days later, except for the negative control group, rats in each group are intraperitoneally injected with KA (15 mg / kg / i.p. / once). Note that the dosage of KA injection is based on previous studies. The seizure behavior of the rats is analyzed within 3 hours after the intraperitoneal injection of KA, and the results are shown in Figure 2.

[0066] As shown in Figure 2, compared with the KA group, the high-dose group (300 mg / kg) of the crude drug composition of the present invention has a significant effect (p < 0.001). Also, the number of animals that did not have epileptic seizures after treatment with the high-dose group of the crude drug composition was the largest (77%, p < 0.001), followed by the medium-dose group (200 mg / kg) of the crude drug composition (33%, p > 0.05) or the low-dose group (100 mg / kg) of the crude drug composition (40%, p > 0.05). In addition, the high-dose group of the crude drug composition is similar (p > 0.05) to 100 mg / kg CBZ in terms of the time of epileptic seizures and the influence on epilepsy evaluation. Such results prove that the crude drug composition of the present invention has an anti-epileptic effect against KA. In other words, the crude drug composition of the present invention can be used as an alternative to anti-epileptic drugs.

[0067] Furthermore, in the anti-epileptic drug composition group (100 mg / kg of crude drug composition + 50 mg / kg of CBZ), the time and degree of animal epileptic seizures are significantly improved (p < 0.05) compared with the KA (15 mg / kg) group. In other words, the anti-epileptic drug composition can greatly improve the drawback that low-dose anti-epileptic drugs cannot effectively suppress epileptic symptoms. Since the anti-epileptic drug composition group has an anti-epileptic effect against KA similar to that of the anti-epileptic drug group, the results show that combining a low-dose (100 mg / kg) of the crude drug composition and a low-dose (50 mg / kg) of CBZ can prevent KA-induced animal epilepsy, and the low-dose CBZ can further reduce the side effects of anti-epileptic drugs.

[0068] As demonstrated in the rat experiments, the method provided by the present invention can prevent epileptic seizures and has a significant preventive and therapeutic effect on particularly intractable epilepsy symptoms. When the herbal medicine composition of the present invention is used alone, an anti-epileptic effect equivalent to that of an anti-epileptic drug (e.g., CBZ) can be obtained. In other words, the herbal medicine composition can be used instead of an anti-epileptic drug. Further, when the herbal medicine composition of the present invention and an anti-epileptic drug are used in combination, an anti-epileptic effect can be obtained and the dosage of the anti-epileptic drug (e.g., CBZ) can be reduced. Therefore, whether the herbal medicine composition of the present invention is used alone or in combination with an anti-epileptic drug, the side effects of the anti-epileptic drug can be reduced. Since the method provided by the present invention has low toxicity and side effects, it becomes one of the safe and effective epilepsy prevention methods.

[0069] [Evaluate the effect of the herbal medicine composition on animal epileptic behavior induced by Example 4-KA by electroencephalogram recording] Divide the rats into 3 groups as in the above example, and give physiological saline to the control group with a dropper at a fixed time every day, give the herbal medicine composition to the herbal medicine composition group, give carbamazepine to the anti-epileptic drug group, and give the herbal medicine composition and carbamazepine to the anti-epileptic drug composition group. After 7 days, except for the negative control group, inject KA (15 mg / kg / i.p. / once) intraperitoneally into the rats in each group. Note that the dosage of KA injection is based on previous research. Analyze the electroencephalogram within 3 hours after intraperitoneal injection of KA for epileptic seizures.

[0070] Electroencephalogram (EEG) recording is performed using a 3-channel EEG system (Pinnacle Technology Inc., Lawrence, Kansas, USA) based on the above method (see references). After anesthetizing the rats with 3% isoflurane, they are fixed in a stereotaxic frame (RWD, Life.Science, Dover, USA), and four electrodes are implanted surgically. Two of the electrodes are placed on both sides of the prefrontal cortex of bregma (anteroposterior position (AP), +3.9; medial (M) ±2.0), and the remaining two are placed in the parietal cortex (AP - 6.4, ML ±4.0). The surgery is performed under isoflurane anesthesia. One week after the surgery, the rats are connected to an EEG amplifier (mode #8213 - SE3), and further, a data acquisition system (Pinnacle Technology Inc., Lawrence, Kansas, USA) is used to record the EEG signals for 3 hours. The electrical signals from the brain are pre-amplified by an 8-channel explorer (HS-8-CNR-MDR50, Neuralynx). Then, the signals are digitized at 1.6 kHz, and further, online band-pass filtering is recorded between 0.1 and 100 Hz by Cheetah 6.2.0 recording software. In the determined recordings, the criteria for epileptic seizures are high-amplitude, rhythmic discharges, including repetitive peaks, peak-wave discharges, and slow waves, and the duration is at least 10 seconds. The PAL-8200EEG software (Pinnacl Technologies) is used to analyze the number and duration of epileptic seizures in the EEG.

[0071] The seizure peak of the rat brain is evaluated by electroencephalogram recording. As shown in FIGS. 3A and 3B, compared with the control group, treatment with KA increased the number and duration of seizure peaks. However, compared with the KA group, the number and duration of seizures in the NRICM101 300 mg / kg + KA group (high-dose crude drug composition group) and the NRICM101 100 mg / kg + CBZ 50 mg / kg + KA group (antiepileptic drug composition group) were decreased, and this effect was also similar to that of the CBZ 100 mg / kg + KA group (antiepileptic drug group) (p>0.05). From these results, both the crude drug composition of the present invention and the combination of the crude drug composition and an antiepileptic drug have the effect of preventing epilepsy induced by KA. Also, when the crude drug composition of the present invention is combined with an antiepileptic drug, it has an antiepileptic effect and can also reduce the dosage of the antiepileptic drug (such as CBZ).

[0072] Based on the results of the above animal experiments, since the crude drug composition of the present invention can extend the time for suppressing epilepsy induced by KA and can reduce the degree of epilepsy induced by KA, it can be proved that the crude drug composition has the effect of preventing epilepsy. Thereby, the crude drug composition of the present invention is used for the development of antiepileptic drugs and / or related products.

[0073] The following provides exemplary implementation methods, and the numbering should not be construed as a level of importance.

[0074] Example 1 provides the use of a crude drug composition as a pharmaceutical for preventing or treating epileptic seizures, and the crude drug composition includes saffron, houttuynia cordata, angelica, calonyction aculeatum, perilla frutescens, cinnamon, peppermint, schizonepeta tenuifolia, bupleurum chinense, and licorice.

[0075] Example 2 provides the use of Example 1. The crude drug composition contains 3.75 to 1.25 parts by weight of saffron, 3.75 to 1.25 parts by weight of houttuynia, 3.75 to 1.25 parts by weight of atractylodes lancea, 3.75 to 1.25 parts by weight of carthamin, 2.25 to 0.75 parts by weight of schizonepeta, 2.25 to 0.75 parts by weight of cinnamon, 2.25 to 0.75 parts by weight of peppermint, 2.25 to 0.75 parts by weight of schizonepeta tenuifolia, 1.5 to 0.5 parts by weight of bupleurum, and 1.5 to 0.5 parts by weight of licorice.

[0076] Example 3 provides the use of Example 1. The crude drug composition contains 2.75 to 2.25 parts by weight of saffron, 2.75 to 2.25 parts by weight of houttuynia, 2.75 to 2.25 parts by weight of atractylodes lancea, 2.75 to 2.25 parts by weight of carthamin, 1.65 to 1.35 parts by weight of schizonepeta, 1.65 to 1.35 parts by weight of cinnamon, 1.65 to 1.35 parts by weight of peppermint, 1.65 to 1.35 parts by weight of schizonepeta tenuifolia, 1.1 to 0.9 parts by weight of bupleurum, and 1.1 to 0.9 parts by weight of licorice.

[0077] Example 4 provides the use of Example 1. The crude drug composition contains 2.5 g of saffron, 2.5 g of houttuynia, 2.5 g of atractylodes lancea, 2.5 g of carthamin, 1.5 g of schizonepeta, 1.5 g of cinnamon, 1.5 g of peppermint, 1.5 g of schizonepeta tenuifolia, 1 g of bupleurum, and 1 g of licorice.

[0078] Example 5 provides the use of Example 1. Epileptic seizures are caused by damage to brain nerve cells.

[0079] Example 6 provides the use of Example 1. Epileptic seizures are caused by the activation of glial cells.

[0080] Example 7 provides the use of Example 1. Epileptic seizures are caused by an inflammatory reaction in the brain triggered by inflammatory molecules.

[0081] Example 8 provides the use of Example 5, and the inflammatory molecules are selected from the group consisting of interleukin 1β (IL-1β), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), high mobility group protein (HMGB1), interleukin 1 receptor 1 (IL-1R1), and toll-like receptor 4 (TLR-4).

[0082] Example 9 provides the use of Example 1, and further at least one antiepileptic drug is combined with the crude drug composition to produce a drug for preventing or treating epileptic seizures.

[0083] Example 10 provides the use of Example 9, and at least one antiepileptic drug is selected from the group consisting of phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepine, tiagabine, and vigabatrin.

[0084] Example 11 provides the use of Example 10, and at least one antiepileptic drug is carbamazepine.

[0085] Example 12 provides the use of Example 1, and the pharmaceutical is used for the treatment and prevention of epilepsy in humans.

[0086] Example 13 provides the use of Example 1, and the pharmaceutical is used for the treatment and prevention of epilepsy in mammals other than humans.

[0087] Example 14 provides a combination of pharmaceuticals for preventing or treating epileptic seizures, including one crude drug composition and at least one antiepileptic drug. The crude drug composition includes saffron, houttuynia cordata, ginseng, kalopanax pictus, magnolia officinalis, cinnamomum cassia, peppermint, schizonepeta tenuifolia, bupleurum chinense, and licorice.

[0088] Example 15 provides a combination of pharmaceuticals of Example 14, and the crude drug composition includes 3.75 to 1.25 parts by weight of saffron, 3.75 to 1.25 parts by weight of houttuynia, 3.75 to 1.25 parts by weight of schizonepeta, 3.75 to 1.25 parts by weight of carthamin, 2.25 to 0.75 parts by weight of perilla, 2.25 to 0.75 parts by weight of cinnamon, 2.25 to 0.75 parts by weight of peppermint, 2.25 to 0.75 parts by weight of schizandra, 1.5 to 0.5 parts by weight of atractylodes lancea, and 1.5 to 0.5 parts by weight of licorice.

[0089] Example 16 provides a combination of pharmaceuticals of Example 14, and the crude drug composition includes 2.75 to 2.25 parts by weight of saffron, 2.75 to 2.25 parts by weight of houttuynia, 2.75 to 2.25 parts by weight of schizonepeta, 2.75 to 2.25 parts by weight of carthamin, 1.65 to 1.35 parts by weight of perilla, 1.65 to 1.35 parts by weight of cinnamon, 1.65 to 1.35 parts by weight of peppermint, 1.65 to 1.35 parts by weight of schizandra, 1.1 to 0.9 parts by weight of atractylodes lancea, and 1.1 to 0.9 parts by weight of licorice.

[0090] Example 17 provides a combination of pharmaceuticals of Example 14, and the crude drug composition includes 2.5 g of saffron, 2.5 g of houttuynia, 2.5 g of schizonepeta, 2.5 g of carthamin, 1.5 g of perilla, 1.5 g of cinnamon, 1.5 g of peppermint, 1.5 g of schizandra, 1 g of atractylodes lancea, and 1 g of licorice.

[0091] Example 18 provides a combination of pharmaceuticals of Example 14, and the dosage of the antiepileptic drug is less than the dosage when the antiepileptic drug is used alone in preventing or treating epileptic seizures.

[0092] Example 19 provides a combination of pharmaceuticals of Example 14, and at least one antiepileptic drug is selected from the group consisting of a combination of phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepine, tiagabine, and vigabatrin.

[0093] Example 20 provides a combination of pharmaceuticals of Example 14, and at least one antiepileptic drug is carbamazepine.

[0094] Example 21 provides a combination of pharmaceuticals of Example 14, and the crude drug composition and at least one anti-epileptic drug are administered to a patient simultaneously, sequentially, or at intervals.

[0095] Example 22 provides a method for preventing or treating epileptic seizures, which includes administering a crude drug composition to an epileptic patient. The crude drug composition includes Curcuma longa, Houttuynia cordata, Bupleurum falcatum, Carthamus tinctorius, Artemisia princeps, Cinnamomum cassia, Mentha piperita, Prunella vulgaris, Pueraria thomsonii, and Glycyrrhiza glabra.

[0096] Example 23 provides the method of Example 22, and the crude drug composition includes 3.75 - 1.25 parts by weight of Curcuma longa, 3.75 - 1.25 parts by weight of Houttuynia cordata, 3.75 - 1.25 parts by weight of Bupleurum falcatum, 3.75 - 1.25 parts by weight of Carthamus tinctorius, 2.25 - 0.75 parts by weight of Artemisia princeps, 2.25 - 0.75 parts by weight of Cinnamomum cassia, 2.25 - 0.75 parts by weight of Mentha piperita, 2.25 - 0.75 parts by weight of Prunella vulgaris, 1.5 - 0.5 parts by weight of Pueraria thomsonii, and 1.5 - 0.5 parts by weight of Glycyrrhiza glabra.

[0097] Example 24 provides the method of Example 22, and the crude drug composition includes 2.75 - 2.25 parts by weight of Curcuma longa, 2.75 - 2.25 parts by weight of Houttuynia cordata, 2.75 - 2.25 parts by weight of Bupleurum falcatum, 2.75 - 2.25 parts by weight of Carthamus tinctorius, 1.65 - 1.35 parts by weight of Artemisia princeps, 1.65 - 1.35 parts by weight of Cinnamomum cassia, 1.65 - 1.35 parts by weight of Mentha piperita, 1.65 - 1.35 parts by weight of Prunella vulgaris, 1.1 - 0.9 parts by weight of Pueraria thomsonii, and 1.1 - 0.9 parts by weight of Glycyrrhiza glabra.

[0098] Example 25 provides the method of Example 22, and the crude drug composition includes 2.5 g of Curcuma longa, 2.5 g of Houttuynia cordata, 2.5 g of Bupleurum falcatum, 2.5 g of Carthamus tinctorius, 1.5 g of Artemisia princeps, 1.5 g of Cinnamomum cassia, 1.5 g of Mentha piperita, 1.5 g of Prunella vulgaris, 1 g of Pueraria thomsonii, and 1 g of Glycyrrhiza glabra.

[0099] Example 26 provides the method of Example 22, and further includes administering at least one anti-epileptic drug.

[0100] Example 27 provides the method of Example 26, and at least one type of antiepileptic drug is selected from the group consisting of phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepine, tiagabine, and vigabatrin in combination.

[0101] Example 28 provides the method of Example 26, and at least one type of antiepileptic drug is carbamazepine.

[0102] Example 29 provides the method of Example 26, and the crude drug composition and at least one type of antiepileptic drug are administered to a patient simultaneously, sequentially, or at intervals.

[0103] Example 30 provides the method of Example 22 or 26, and the patient is a mammal.

[0104] Example 31 provides the method of Example 30, and the patient is a human.

[0105] Although the present invention discloses certain embodiments, those skilled in the art can design other embodiments and modifications without departing from the spirit and scope of the present invention. The claims cover modifications having the same effect as any of the embodiments.

Industrial Applicability

[0106] The crude drug composition provided by the present invention can firmly suppress epileptic seizures and reduce the degree of epilepsy. Furthermore, by combining with other antiepileptic drugs, the antiepileptic effect can be enhanced and the side effects of antiepileptic drugs can be reduced. Accordingly, the crude drug composition of the present invention is used in the development of antiepileptic drugs and / or related products.

Claims

1. Use of a crude drug composition for preventing or treating epileptic seizures, wherein the crude drug composition contains Scutellaria baicalensis, Houttuynia cordata, Isatis indigotica, Trichosanthes kirilowii, Morus alba, Magnolia officinalis, Mentha haplocaly x, Nepeta tenuifolia, Saposhnikovia divaricata and Glycyrrhiza glabra. Use of the crude drug composition characterized by this.

2. The epilepsy according to claim 1, characterized in that it is caused by damage to brain nerve cells. Use of the crude drug composition described in claim 1.

3. The epilepsy according to claim 1, characterized in that it is caused by activation of glial cells. Use of the crude drug composition described in claim 1.

4. The epilepsy according to claim 1, characterized in that it is caused by an inflammatory reaction in the brain caused by inflammatory molecules. Use of the crude drug composition described in claim 1.

5. The inflammatory molecules are selected from the group consisting of a combination of Interleukin-1β (IL-1β), Interleukin-6 (IL-6), Tumor Necrosis Factor-α (TNF-α), High Mobility Group Box 1 (HMGB1), Interleukin-1 Receptor 1 (IL-1R1), and Toll-like Receptor 4 (TLR-4). Use of the crude drug composition described in claim 4, characterized by this.

6. At least one anti-epileptic drug is combined with the crude drug composition to produce a drug for preventing or treating epileptic seizures. Use of the crude drug composition described in claim 1, characterized by this.

7. The use of the crude drug composition according to claim 1, wherein at least one type of antiepileptic drug is selected from the group consisting of phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepines, tiagabine, and vigabatrin.

8. A pharmaceutical combination for preventing or treating epileptic seizures, comprising a crude drug composition and at least one type of antiepileptic drug, wherein the crude drug composition comprises saffron, houttuynia, ginseng, carthamus, perilla, cinnamon, peppermint, gardenia, bupleurum, and licorice, and the dosage of the antiepileptic drug is less than the dosage when the antiepileptic drug is used alone in preventing or treating epileptic seizures.

9. The pharmaceutical combination according to claim 8, wherein at least one type of antiepileptic drug is selected from the group consisting of phenytoin, carbamazepine, lamotrigine, felbamate, benzodiazepines, tiagabine, and vigabatrin.

10. The pharmaceutical combination according to claim 8, wherein the crude drug composition and at least one type of antiepileptic drug are administered to a patient simultaneously, sequentially, or at intervals.

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