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WO2015072667A1 - Composition pharmaceutique pour prévenir ou traiter des maladies nerveuses crâniennes neurodégénératives - Google Patents

Composition pharmaceutique pour prévenir ou traiter des maladies nerveuses crâniennes neurodégénératives Download PDF

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Publication number
WO2015072667A1
WO2015072667A1 PCT/KR2014/009632 KR2014009632W WO2015072667A1 WO 2015072667 A1 WO2015072667 A1 WO 2015072667A1 KR 2014009632 W KR2014009632 W KR 2014009632W WO 2015072667 A1 WO2015072667 A1 WO 2015072667A1
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treatment
composition
prevention
pharmaceutical composition
present
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Korean (ko)
Inventor
김선림
정건호
김미정
이유영
김기종
손범영
이진석
김정태
백성범
권영업
박용일
최두진
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Industry Academic Cooperation Foundation of Catholic University of Korea
Korea Rural Development Administration
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Industry Academic Cooperation Foundation of Catholic University of Korea
Korea Rural Development Administration
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2121/00—Preparations for use in therapy

Definitions

  • the present invention relates to a pharmaceutical composition for preventing or treating neurodegenerative neurodegenerative diseases, and more particularly, to prevent or treat neurodegenerative neurodegenerative diseases and to prevent neurodegenerative neuroprotective effects. It is about.
  • Neurodegenerative disease is known to be caused by the death of neurons due to oxidative stress. Recently, degenerative brain neurological diseases including Alzheimer's disease have been cited as the main cause of the disease caused by oxidative stress due to the rapid increase in the amount of free radicals in neurons. Therefore, it is reported that such diseases can be prevented or treated by suppressing or reducing oxidative stress.
  • Intracellular antioxidant activity consists of an enzymatic defense system and an antioxidant system.
  • Enzymatic defense system protects against oxidative stress by enzymes such as catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), and heme oxygenase (HO). It is a mechanism to defend. In particular, HO has been reported to play an important role in maintaining homeostasis through antioxidant activity in the oxidative state.
  • CAT catalase
  • GPx glutathione peroxidase
  • SOD superoxide dismutase
  • HO heme oxygenase
  • Corn beard has traditionally been ingested as a edible or folk remedy for various diseases, has a relatively high stability, and has a high potential of development as a new drug derived from natural products.
  • Korean Patent No. 10-1201628 the pharmacological effects of the antimicrobial and anticancer effects of corn beard extracts have been studied, but there has been no report on the inhibitory activity and the neuroprotective activity against neuronal cell death caused by oxidative stress.
  • H 2 O 2 By reducing the oxidative stress induced by H 2 O 2 , there was a problem that a composition for improving neurodegenerative diseases due to inhibition of neuronal cell death has not been developed.
  • the present invention has been made to solve the above problems, the first problem to be solved of the present invention is to provide a pharmaceutical composition for the prevention or treatment of neurodegenerative diseases of the neurons to inhibit apoptosis.
  • a second object of the present invention is to provide a dietary supplement for the prevention or improvement of neurodegenerative diseases comprising the composition.
  • the present invention provides a pharmaceutical composition for the prevention or treatment of degenerative cerebral nervous system diseases containing mayin as an active ingredient in order to achieve the first object.
  • the composition can inhibit neuronal cell death.
  • the composition may be derived from corn beard.
  • the composition may increase the activity of antioxidant enzymes of neurons.
  • the composition can inhibit the oxidative stress induced by H 2 O 2 .
  • the composition can inhibit reactive oxygen species of neurons.
  • the composition may exhibit a protective effect of neurons.
  • the composition may be characterized in that it is contained at a concentration of 5 ⁇ 100 ⁇ g / ml.
  • the degenerative neurological disease may include Alzheimer's, Parkinson's disease, Huntington's disease, multiple sclerosis, multiple neurotrophic, epilepsy, brain disease (encephalopathy), or stroke.
  • the second object provides a health functional food composition for the prevention or improvement of neurodegenerative diseases containing the composition.
  • prevention refers to any action that inhibits apoptosis of neurons or delays apoptosis of neurons by administration of a composition
  • treatment refers to symptoms caused by onset by administration of the composition. Means any action that improves or beneficially changes;
  • the present invention reduces the amount of reactive oxygen species by effectively inhibiting the activity of oxidative stress, and has an excellent protective effect against the damage of activated oxidative stress on nerve cells, which can be usefully used as an effective preventive or therapeutic agent for degenerative cerebral neuropathy. have.
  • Figure 1a is an experimental result of neuronal cell death by treatment of the mycin according to a preferred embodiment of the present invention, the figure showing the effect of inhibiting apoptosis of neurons using Annexin V / PI fluorescence staining.
  • FIG. 1B is a graph showing numerical results of using the Annexin V / PI fluorescence stain as an experimental result of neuronal cell death by treatment with Meisin according to a preferred embodiment of the present invention.
  • Figure 2 is an experimental result of inhibiting DNA fragmentation by the treatment of the mycin according to an embodiment of the present invention, showing the DNA fragmentation induced by H 2 O 2 using ApoDIRECT In Situ DNA fagmentation assay kit fluorescent staining Picture.
  • Figure 3 is a graph showing the experimental results of inhibiting the activity of reactive oxygen species by the treatment of the mycin according to an embodiment of the present invention.
  • Figure 4a is a graph showing the results of the experiment observed the expression of catalase (CAT) which is an antioxidant enzyme in neurons by treatment of the mycin according to a preferred embodiment of the present invention.
  • CAT catalase
  • Figure 4b is a graph showing the results of the experiment observed the expression of glutathione peroxidase-1 (GPx-1), an antioxidant enzyme in neurons by the treatment of the mycin according to a preferred embodiment of the present invention.
  • GPx-1 glutathione peroxidase-1
  • Figure 4c is a graph showing the results of the experiment observed the expression of superoxide dismutase-1 (SOD-1), an antioxidant enzyme in neurons by treatment of the mycin according to a preferred embodiment of the present invention.
  • SOD-1 superoxide dismutase-1
  • Figure 4d is a graph showing the results of the experiment observed the expression of superoxide dismutase-2 (SOD-2), an antioxidant enzyme in neurons by the treatment of the mycin according to a preferred embodiment of the present invention.
  • SOD-2 superoxide dismutase-2
  • Figure 4e is a graph showing the results of the experimental observation of the increased expression of the antioxidant enzyme heme oxygenase-1 (HO-1) in neurons by the treatment of the mycin according to an embodiment of the present invention.
  • HO-1 heme oxygenase-1
  • Figure 5a is a graph showing the experimental results of observing the increase in the cell survival rate of neurons by the treatment of the mycin according to a preferred embodiment of the present invention.
  • Figure 5b is a graph showing the results of the experiments observed that the cytotoxicity of neurons is reduced by the treatment of the mycin according to an embodiment of the present invention.
  • a pharmaceutical composition for the prevention or treatment of degenerative brain neurological disease containing mayin as an active ingredient. This effectively inhibits the cell death of nerve cells by effectively inhibiting the activity of oxidative stress, thereby excellent protection against neuronal cell damage caused by oxidative stress can be useful as an effective preventive or therapeutic agent of degenerative neurological diseases. have.
  • the present invention includes the drug as an active ingredient.
  • Maysin is a yellow plant pigment flavonoid family found in corn, teosinte and centipedes grass, and has been known for its antioxidant and anticancer activity.
  • the present invention confirmed the prevention and treatment effect of maize mustache-derived degenerative neurological disease.
  • Meisin of the present invention is derived from corn beard, according to a preferred embodiment of the present invention, it can provide a composition for improving neurodegenerative disorders of the brain, which comprises a mysin derived from corn beard as an active ingredient.
  • a novel physiological activity of macine derived from the cornbeard was developed. As a result, it was confirmed that the macine inhibited the neuronal cell death by oxidative stress, and the neurons were protected from the oxidative stress by increasing the expression of antioxidant enzymes.
  • the active ingredient of the present invention may inhibit the cell death of neurons by increasing the expression of antioxidant enzymes in neurons.
  • Example 1 of the present invention it was confirmed that apoptosis of human neuroblastoma (SK-N-MC) induced by H 2 O 2 was inhibited by treating the mycin.
  • SK-N-MC human neuroblastoma
  • Example 2 of the present invention the macine inhibits apoptosis of neurons induced by H 2 O 2 and inhibits DNA fragmentation activity by apoptosis. You can check it.
  • apoptosis of the present invention is a programmed apoptosis process at a certain time and place, and many human diseases as well as physiological processes such as tissue homeostasis and embryonic development. Excessive cell death leads to atrophy and degenerative neuropathy.
  • the may be able to inhibit the oxidative stress induced by H 2 O 2 .
  • Our body regulates the amount of free radicals in the body by itself. When excessive exercise is excessively increased due to excessive exercise, or the ability to remove them, various diseases caused by harmful oxygen are caused. The side effects of the harmful oxygen is called oxidative stress. Overproduction of free radicals, which causes oxidative stress to accumulate continuously in the body, affects the genes of cells or damages the immune system.
  • the macine inhibits reactive oxygen species (ROS) by inhibiting oxidative stress.
  • ROS reactive oxygen species
  • the reactive oxygen species overproduced by H 2 O 2 refers to an unstable state having free radicals, and thus has strong activity.
  • H 2 O 2 refers to an unstable state having free radicals, and thus has strong activity.
  • Example 3 of the present invention the active oxygen species in the neurons induced by H 2 O 2 was found to be reduced by the treatment of Meissin, H 2 O 2 and the control group not treated with H 2 O was analyzed by comparing the second one nerve cell process, the concentration of the active oxygen species of the H 2 O 2 treated neurons was confirmed that the increase of 1.62 times compared with the control group. In addition, when treated with 10 ⁇ g / ml of macine in neurons, it was confirmed that the concentration of reactive oxygen species decreased by 40 to 45%. It was found that the activity of reactive oxygen species is mediated by H 2 O 2 .
  • the intracellular antioxidant action is composed of an enzymatic defense system and an antioxidant system. Therefore, antioxidant enzymes are very important in inhibiting oxidative stress in cells.
  • the macine may confirm that the activity of antioxidant enzymes in neurons is increased.
  • the antioxidant enzyme may act as a biocatalyst. Biocatalysts, such as catalase, are included in the animal and plant kingdoms, decomposing harmful hydrogen peroxide from metabolic processes into oxygen, and providing oxygen back to oxidation.
  • Example 4 of the present invention it was confirmed that the enzymes such as catalase, glutathione peroxidase, superoxide dismutase and heme oxygenase increased by 2.49 to 14.75 times by treating the neurons with neurons. More specifically, catalase (CAT) increased the enzyme activation by 260-265% when 25 ⁇ g / ml of macine was treated, and glutathione peroxidase (GPx-1) treated 25 ⁇ g / ml of macine In this case, it was confirmed that the activation of the enzyme increased by 200 to 205%. Also.
  • catalase CAT
  • glutathione peroxidase GPx-1
  • Superoxide dismutase-1 (SOD-1) increased the enzyme activation by 260 ⁇ 265% when 25 ⁇ g / ml of macine was treated
  • superoxide dismutase-2 (SOD-2) increased 25 ⁇ g / ml of macine
  • the activation of enzyme was increased by 230-235%
  • heme oxygenase (HO-1) was found to increase the activation of enzyme by 550-555% when 10 ⁇ g / ml of mesine was treated.
  • These enzymes are a mechanism that transforms free radicals produced by oxidative stress in nerve cells into stable substances and protects nerve cells from damage caused by oxidative stress through metabolic processes.
  • the macine protects the cells from damage of neurons by inhibiting oxidative stress in neurons.
  • the concentration of the macine is 0.1 ⁇ 1000 ⁇ g / ml, preferably 1.0 ⁇ 500 ⁇ g / ml Concentrations, more preferably at a concentration of 7.5-100 ⁇ g / ml, can achieve the physiological effects described above.
  • the inhibitory effect on the generation of reactive oxygen species was remarkable between the concentrations of Mayine between 5.0 ⁇ g / ml and 10.0 ⁇ g / ml. Therefore, it can be seen that most preferably, when the concentration of macine is 7.5 ⁇ g / ml, there is a significant difference in effect.
  • the macine increases cell viability and decreases cytotoxicity.
  • the cell viability is increased according to the concentration of macine, and the cytotoxicity of the control group is not treated with H 2 O 2 .
  • the toxicity of the treatment with H 2 O 2 cells compared to the control group not treated with H 2 O 2 4 times as it was confirmed that the number of toxicity. It was confirmed that the mycin is mediated in preventing cytotoxicity caused by H 2 O 2 .
  • the neurodegenerative diseases of the present invention may be Alzheimer's, Parkinson's disease, Huntington's disease, multiple sclerosis, multiple neurotrophic, epilepsy, brain disease (encephalopathy), or stroke.
  • degenerative brain neuropathy means a term that generically refers to various diseases related to nerves, in particular, cranial nerves.
  • the degenerative brain disease according to the present invention means diseases which the nerve cell damage caused by the oxidative stress induced by H 2 O 2 induced.
  • composition for preventing or treating degenerative neurological disease of the present invention may include a pharmaceutically acceptable carrier.
  • the composition comprising a pharmaceutically acceptable carrier may be in various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
  • Solid form preparations for oral administration include tablet pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose in one or more compounds. ) And gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used.
  • Liquid preparations for oral administration include suspensions, solution solutions, emulsions, and syrups, and various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • non-aqueous solvent and the suspension solvent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used.
  • base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • the pharmaceutical composition is any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, liquid solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations and suppositories. It can have one formulation.
  • compositions are useful for administering TSPOs of the invention to a subject for the treatment of various diseases, including neurodegeneration and / or symptoms associated therewith, as described above.
  • composition of the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level is determined by the type and severity of the subject, age, sex, activity of the drug, drug Sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of drugs, and other factors well known in the medical arts.
  • compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple doses. In consideration of all the above factors, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects.
  • Typical dosages of the pharmaceutical compositions of the invention are in the range of 0.001-100 mg / kg on an adult basis.
  • the route of administration of the pharmaceutical composition may be administered via any general route as long as it can reach the target tissue.
  • the composition of the present invention may be administered as desired, but is not limited to intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, intranasal administration, pulmonary administration, rectal administration.
  • the composition may also be administered by any device in which the active agent may migrate to the target cell.
  • composition of the present invention can be used alone or in combination with methods using surgery, hormonal therapy, drug treatment and biological response modifiers for the prevention and treatment of neurodegenerative diseases.
  • a health functional food composition for the prevention or improvement of neurodegenerative diseases comprising the composition.
  • the mycin of the present invention may be added as it is or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method.
  • the mixed amount of the active ingredient can be determined suitably according to the purpose of use (prevention, health or therapeutic treatment).
  • the term "health functional food” refers to a food prepared and processed by extracting, concentrating, refining, and mixing a specific ingredient as a raw material or contained in a food ingredient for the purpose of health supplement. It refers to foods that are designed and processed to sufficiently exert bioregulatory functions on the living body, such as biological defense, regulation of biorhythms, prevention and recovery of diseases, by the above components, and the composition for health foods prevents diseases and It can perform functions related to the recovery of diseases.
  • tofu extract of the present invention can be prepared by mixing the known additives with other appropriate auxiliary ingredients that may be contained in the health functional food according to the choice of those skilled in the art.
  • foods that can be added include meat, sausages, breads, chocolates, candy, snacks, confections, pizzas, ramen noodles, dairy products including other noodles, gums, ice creams, various soups, beverages, teas, drinks, alcoholic beverages and Vitamin complexes, and the like, can be prepared by adding the extract according to the present invention as a main ingredient juice, tea, jelly and juice.
  • Example 1-1 H 2 O 2 On by Induction of Apoptosis of Neurons
  • Apoptosis was measured using the Annexin V-FITC apoptosis detection kit (Bio Vision, USA). Neurons (SK-N-MC) were incubated for 24 hours in a 6-well plate and 5, 10, 25, 50 ⁇ g / ml of Meisin was pretreated with the cells for 2 hours, followed by H 2 O 2 for 24 hours. Treated. Cells were collected by treatment with Trypsin-EDTA, centrifuged to remove supernatant, suspended in 500 ⁇ l binding buffer, and 5 ⁇ l Annexin V-FITC and 5 ⁇ l PI (propidium iodide) were added. The reaction was carried out for 5 minutes with the light blocked at room temperature. Analysis was performed using flow cytometry.
  • apoptotic apoptosis was increased by about 8.05 times, and apoptosis was reduced by 85-90% when pretreated with 50 ⁇ g / ml of macine (FIGS. 1A and 1B). ).
  • SK-N-MC a human blastoma
  • H 2 O 2 treatment and fluorescence using Annexin V / PI After staining, cell death was observed using a flow cytometer.
  • Figure 1A and Figure 1B as a result of Example 1, the active ingredient of the macine was able to confirm the neuronal cell death inhibitory effect.
  • control refers to a control group not treated with both Meisin and H 2 O 2 .
  • H 2 O 2 was treated alone was shown. It is a graph showing that the treatment of the micelle with H 2 O 2 and concentration.
  • FIG. 1B quantifying the results of FIG. 1A, it can be seen that there is a significant difference in effect when the concentration of the macine is 5 to 10 ⁇ g / ml.
  • H 2 O 2 is produced by disproportionation of superoxide anion and various oxidases in vivo, and is also a representative compound of oxygen poison. Since it is removed by the antioxidant enzyme in vivo, it was confirmed that the oxidase was removed by the antioxidant enzyme of the macine to confirm the effect of inhibiting cell death of neurons.
  • the DNA fragmentation means DNA fragmentation, and when apoptosis occurs in a cell, DNA fragmentation separated from the nucleus can be confirmed little by little.
  • the cells shrink, gaps are created between them, and DNA is regularly cut and fragmented within the cells. At the end, the entire cell is fragmented and eaten by adjacent cells, ending their lifetime. Therefore, it can be seen that inhibiting cell apoptosis by inhibiting DNA fragmentation.
  • comtrol refers to a control group not treated with both macine and H 2 O 2 .
  • H 2 O 2 concentration By H 2 O 2 concentration and a diagram showing that the treatment with god mate. As shown in the figure, DNA fragmentation decreases as the concentration of macine increases.
  • the active ingredient of mayin can be usefully used to suppress the cell death of neurons by effectively inhibiting DNA fragmentation that causes cell death.
  • Example 3-1 H 2 O 2 in Free radical species induction
  • ROS reactive oxygen species
  • Active oxygen species refers to a state in which free radicals are not stable because they have free radicals, and thus have strong activity. Excessive production of reactive oxygen species is called toxic oxygen because it causes toxicity to the living organism, that is, oxidative stress. By oxidizing huge molecules (proteins, lipids, etc.) in the cell, it destroys the homeostasis of the cell, kills the cell, and causes fatal damage in the tissue.
  • Reactive oxygen species include singlet oxygen, superoxite radicals, hydroxy radicals and hydrogen peroxide (H 2 O 2 ).
  • Example 3 Through the experimental results of Example 3, it can be confirmed that the macine is effective in inhibiting reactive oxygen species.
  • Example 4-1 Isolation of Total RNA
  • mRNA level of the antioxidant enzymes was examined by real-time PCR.
  • Neurons (SK-N-MC) were incubated for 24 hours in a 6-well plate and treated with mayin at concentrations of 5, 10, 25 and 50 ⁇ g / ml for 2 hours, and then H 2 O 2 for 24 hours. Treated. After the reaction, the solution was washed with PBS, dissolved with TRIzol reagent (invitrogen, USA), and then added with Chloroform (Sigma, USA) and centrifuged at 4 ° C. and 13,000 rpm for 10 minutes.
  • RNA is isolated using Total RNA Extraction kit (intron, Korea). After the reaction by adding 400 ⁇ L of Binding Buffer to the isolated RNA, wash using washing buffer A and B. Finally, 50 ⁇ l of RNA is eluted with elution buffer. Total RNA isolated was quantified with a spectrophotometer to a concentration of 1 ⁇ g.
  • RNA isolated was synthesized first strand complementary DNA (cDNA) using the Power cDNA Synthesis Kit (intron, Korea). 1 ⁇ g of Oligo (dT) 15primer was added to 1 ⁇ g of total RNA, followed by reaction at 75 ° C. for 5 minutes, followed by reaction at 4 ° C. for 1 minute. Add RNase inhibitor, 5X RT buffer, dNTP, DTT, AMV RT enzyme and react for 1 hour at 42 °C. CDNA obtained by reacting for 5 minutes at 75 °C was used for real-time PCR.
  • mRNA levels of all antioxidant enzymes were decreased in the H 2 O 2 treated group compared to the control group without any treatment.
  • CAT, GPx-1, SOD-1,2 were treated in the 50 ⁇ g / ml treatment group , HO-1 mRNA level was confirmed to increase 2.49 times, 2.42 times, 2.40 times, 3.15 times, 14.75 times, respectively (Figs. 4A, 4B, 4C, 4D and 5E).
  • the antioxidant enzymes are beneficial substances for removing oxidase present in cells
  • SOD-1 and SOD-2 are antioxidant enzymes that inhibit free radicals by decomposing the over-production of harmful active oxygen into hydrogen peroxide and oxygen.
  • Hydrogen peroxide decomposed by SOD-1 and SOD-2 is decomposed into oxygen and water by catalase and glutathione peroxidase to act as antioxidant enzymes.
  • heme oxygenase is an effective enzyme that maintains homeostasis by oxidizing heme in the blood. Therefore, the macine is useful for effectively removing free radicals by enhancing intracellular antioxidant enzymes.
  • Figure 4A showing the experimental results carried out in Example 4 is a graph quantifying the expression amount of catalase through the level of mRNA. It can be seen that there is a significant difference in the effect between the concentration of 5 ⁇ 10 ⁇ g / ml of mesine, the catalase was expressed up to 80 ⁇ 85% of the control when treated with 50 ⁇ g / ml.
  • Figure 4B showing the experimental results carried out in Example 4 is a graph quantifying the expression amount of glutathione peroxidase-1 through the level of mRNA. It can be seen that the expression amount is increased by 2.42 times when the macine is treated with 50 ⁇ g / ml, compared with the case without the treatment with the mycin.
  • Figure 4C showing the experimental results carried out in Example 4 is a graph quantifying the expression amount of superoxide dismutase-1 through the level of mRNA. When treated with 50 ⁇ g / ml of mayin compared to the case without the treatment, it can be seen that the expression amount increases 2.40 times.
  • Figure 4D showing the experimental results carried out in Example 4 is a graph quantifying the expression amount of superoxide dismutase-2 through the mRNA level. It can be seen that the amount of expression increased by 3.15 times when treated with 50 ⁇ g / ml compared to the case without treatment with the mycin.
  • Figure 4E showing the experimental results carried out in Example 4 is a graph quantifying the expression amount of heme oxygenase through the mRNA level. It can be seen that the expression amount increased by 14.75 times when treated with 50 ⁇ g / ml compared to the case without treatment with the mycin.
  • the change in neuronal toxicity when H 2 O 2 treatment was measured by LDH assay.
  • SK-N-MC cells were incubated in a 6-well plate for 24 hours at 37 ° C. and 5% CO 2 conditions, 5-50 ⁇ g / ml mayin was pretreated for 2 hours, and 200 ⁇ M H 2 O 2 was treated for 24 hours. .
  • the supernatant was taken by centrifugation at 250 ⁇ g, 10 min, 4 °C condition and LDH activity was measured using LDH cytotoxicity detection kit (Takara bio inc, Tokyo, Japan).
  • FIG. 5A showing the results of the experiment conducted in Example 5, it was found that the cell viability was increased by treating the mycin.
  • concentration of 50 ⁇ g / ml it was confirmed that to increase the 80 to 85% of the conrtol group effectively inhibit the H 2 O 2 .
  • Treatment of macine increases the cell viability of neurons.
  • Figure 5B is a graph showing a decrease in cytotoxicity by treatment with macine. When treated with 50 ⁇ g / ml of mayin, it was confirmed that the cytotoxicity is reduced by 60 ⁇ 65% compared to the case without the treatment with the mycin.
  • Example 5 it was confirmed that the cell viability was increased and cytotoxicity was reduced through the treatment of Meicin.
  • tablets were prepared by tableting according to a conventional method for producing tablets.
  • the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.
  • Chewing gum was prepared in a conventional manner by combining 20% by weight of gum base, 70% by weight of sugar, 1% by weight of perfume, 2% by weight of water, and 7% by weight of the above-mentioned macine.
  • Candy was prepared in a conventional manner by combining 50% by weight of sugar, 39.8% by weight of starch syrup, 0.2% by weight of fragrance, and 10% by weight of the mesine.
  • honey 0.26% by weight of honey, 0.0002% by weight of thioctoamide, 0.0004% by weight of nicotinic acid, 0.0001% by weight of sodium riboflavinate, 0.0001% by weight of pyridoxine hydrochloride, 0.001% by weight of inositol, 0.002% by weight of orthoic acid and 89.7362% by weight of water and the may A beverage was prepared by the conventional method by combining 10% by weight of thinner.

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  • Epidemiology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Psychiatry (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Hospice & Palliative Care (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

La présente invention concerne une composition destinée à atténuer les maladies du système nerveux crânien induites par un stress oxydatif, contenant de la maysine dérivée de soie de maïs comme ingrédient actif, et plus particulièrement, une composition pharmacologique destinée à atténuer les maladies du système nerveux crânien dégénératives, la maysine augmentant l'expression des ARNm d'enzymes antioxydantes (catalase, glutathion peroxydase-1, superoxyde dismutases 1 et 2 et hème oxygénase-1), en réduisant ainsi le stress oxydatif et en inhibant ou en bloquant finalement l'apoptose des cellules nerveuse provoquée par le stress oxydatif.
PCT/KR2014/009632 2013-11-18 2014-10-14 Composition pharmaceutique pour prévenir ou traiter des maladies nerveuses crâniennes neurodégénératives Ceased WO2015072667A1 (fr)

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KR1020130139985A KR101554343B1 (ko) 2013-11-18 2013-11-18 퇴행성 뇌신경질환의 예방 또는 치료용 약학적 조성물
KR10-2013-0139985 2013-11-18

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KR20220000069A (ko) * 2020-06-25 2022-01-03 연세대학교 산학협력단 뇌 염증 및 퇴행성 뇌 질환의 예방 또는 치료용 조성물

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KR101201628B1 (ko) * 2011-05-18 2012-11-14 대한민국(농촌진흥청장) 메이신 함량이 높은 옥수수수염 추출물의 제조방법

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CN110251732A (zh) * 2019-06-18 2019-09-20 南通纺织丝绸产业技术研究院 一种多层复合编织的可降解神经导管结构及其制备方法
CN110251732B (zh) * 2019-06-18 2020-10-27 南通纺织丝绸产业技术研究院 一种多层复合编织的可降解神经导管结构及其制备方法

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