Composition for protecting nerve cells containing naphthopyrone derivatives derived from Cassia seed buds

Abstract

The present invention relates to naphthopyrone derivatives and compositions containing Cassia seed bud extracts containing the same as active ingredients. The composition according to one aspect of the present invention has an antioxidant effect, and an effect of protecting nerve cells from oxidative stress or inhibiting nerve cell death, and in particular, has an effect of inhibiting damage or death of retinal nerve cells or hippocampal nerve cells caused by glutamate toxicity, and can be used as a pharmaceutical composition for the treatment or prevention of decreased vision and eye diseases caused by optic nerve damage, and memory loss, learning impairment, depression onset and deterioration, and degenerative brain nerve diseases caused by cranial nerve damage.

Description

Technical Field

[0001] The present specification relates to naphthopyrone derivatives and compositions containing a Cassia seed bud extract containing the same as an active ingredient. Background Art

[0002] The hippocampus, as a structure in the temporal lobe of the brain, plays an important role in the cognitive function of humans and animals. The hippocampus is known to be a central tissue that is vulnerable to physiological / oxidative stress stimulation and can cause damage to cognitive function caused by stress. Stress can cause changes in the structure and brain cell generation, synaptic plasticity, and behavior related to the hippocampus (Eun-Joo Kim, Korean Journal of Psychology: Cognition and Biology, 2012, 24, 65-88). It is known that when oxidative stress in nerve cells is induced in the hippocampus, pituitary gland, striatum, substantia nigra, anterior cerebral cortex or hypothalamus, nerve cell death increases, neurons and growth factors are reduced, thereby leading to acute or chronic neurological diseases such as amyotrophic lateral sclerosis (Lou Gehrig's disease), Parkinson's disease, or cerebral ischemic nerve damage (Rahman, T. et al. Adv. Biosci. Biotechnol. 2012, 3, 997-1019).

[0003] According to health insurance data from the National Health Insurance Agency of Korea, the number of patients with retinal diseases increased from about 830,000 in 2010 to about 1.25 million in 2015, an average annual increase of about 8%. The retina is a thin neural membrane inside the eye similar to a camera film. In the retina, there are more than 100 million photoreceptor cells (cells that sense light) and more than one million optic nerve cells, which convert light into electrical signals and transmit the image of things seen with the eyes to the brain through nerves. If such retinal nerves are damaged or nerve function is abnormal, problems will occur in vision and visual field. Representative retinal diseases include decreased vision, visual field impairment and blindness, including diabetic retinopathy, age-related macular degeneration, retinitis pigmentosa, retinal detachment and retinal vascular occlusion. Glaucoma is a representative eye disease caused by damage to the optic nerve of the retina. It damages the optic nerve due to increased intraocular pressure, ischemia, oxidative stress, etc. (Kim, NY et al. J. Korean Opthalmol. Soc. 2015, 56, 70-79; Kang, JH et al. J. Korean Ophthalmol. Soc. 2003, 44, 965-970; Lee, SM et al. J. Korean Ophthalmol. Soc. 2002, 43, 2577-2584).

[0004] Glutamate is an excitatory neurotransmitter that plays an important role in the central nervous system of vertebrates. Glutamate acts on quisqualate receptors, NMDA (N-methyl-D-aspartate, N-methyl-D-aspartate) receptors and kainite receptors that are concentrated in the cerebellar tonsils and hippocampus, which are known to be involved in memory and learning ability in brain tissue, and participates in various physiological effects. On the contrary, when glutamate is free from the inside of nerve cells to the outside and the extracellular glutamate concentration increases sharply, it can act as an oxidative neurotoxic substance (HyunjeongKim et al., J.Life Sci.2009,19,963-967) to play a role. Excessive glutamate is known to be related to various acute neurological disorders including anemia, hypoxia, hypoglycemia, trauma, and various chronic degenerative neurological diseases. Glutamate in the eye is associated with acute disorders and death of retinal ganglion cells (Otori, Y. et al. Invest. Ophthalmol. Vis. Sci. 1998, 39, 972-981). It is known that while glutamate is produced in large quantities and activates presynaptic glutamate receptors, it stimulates the excessive production of reactive oxygen species (ROS) (Tarasenko A. et al. Neurochem. Int. 2012, 61, 1044-1051). In addition, it is known that the oxidative effects exerted in response to glutamate are mediated by the activation of NMDA receptors, showing calcium ion dependence. It is reported that such excessive activation of NMDA receptors and the uptake of calcium ions in mitochondria cause the generation of ROS (Reynolds IJ et al. J. Neurosci. 1995, 15, 3318-3327). Reactive oxygen species (ROS) are produced continuously by the oxidation and reduction of oxygen based on the respiration and immune response in the mitochondria in the cells of organisms. Because ROS is chemically unstable and highly reactive, it can react with lipids, nucleic acids, proteins, etc. in the body, thereby causing DNA damage, increased concentrations of intracellular free calcium and iron, damage to the ion transport system of biomembranes, etc. (Kiselyov, K. et al. Cell Calcium. 2016, 60, 108-114). In addition, it is known that light can also induce ROS production, thereby inducing photooxidative damage of optic nerves (Masuda, T. et al. Oxid. Med. Cell. Longevity. 2017; article ID 9208489, 14 pages).

[0005] It is known that Cassia seed is traditionally used for eye health and has excellent antioxidant effect. Cassia seed is a mature seed of Cassia seed (Cassia obtusifolia L.) or Longgang Nancha (Cassiatora L.) belonging to Leguminosae, and is cultivated in various parts of Korea. After the leaves fall, there is a row of moist seeds in the arched pods of about 10 cm (Yen, G.-C. et al. J. Agric. Food Chem. 1998, 46, 820-824). Studies have reported that Cassia seed extract has the effects of improving diabetes, improving dyslipidemia, protecting the liver, antibacterial, and lowering blood pressure (Dong, X. et al. Mol. Med. Rep. 2017, 16, 2331-2346).

[0006] However, there has been no report on the neuroprotective effects of Cassia seed bud extract or naphthopyrone derivatives thereof, such as 7-hydroxymusizinyl-rubrofusarin-8'-O-glucopyranoside, isotoralactone, toralactone, torosachrysone and rubrofusarin, against glutamate-induced neurotoxicity, and the technology for the antioxidant effect of Cassia seed bud extract and the composition for neuroprotection is not known.

[0007] In addition, there have been no reports on the research and development of health functionality and medicinal ingredients using Cassia seed bud extract. The inventors of the present invention have confirmed that compared with Cassia seed extract, Cassia seed bud extract produces new antioxidant components and increases antioxidant active components, thereby significantly increasing antioxidant effects and neuronal cell protective effects. The present invention was completed by confirming that the naphthopyrone component containing the new compound 7-hydroxysomycin-erythromycin-8'-O-pyranoglucoside isolated from Cassia seed bud extract shows the effect of protecting retinal neurons and hippocampal neurons from glutamate-induced oxidative stress.

[0008] [Prior art literature]

[0009] [Patent Document]

[0010] (Patent Document 1) KR10-1503429B1

[0011] (Patent Document 2) KR10-2010-0082054A

[0012] (Patent Document 3) KR10-0877371B1

[0013] (Patent Document 4) KR10-1807367B1

[0014] (Patent Document 5) KR10-2016-0058613A

[0015] [Non-patent literature]

[0016] (Non-patent document 1) Jung, HA Journal of Ethnopharmacology, 2016, vol191, 152-160; Inhibitory activities of major anthraquinones and other constituents from Cassia obtusifolia against β-secretase and cholinesterases.

[0017] (Non-patent document 2) Shrestha, S. et al. Archives Pharmacal Research, 2018, online publication https: / / doi.org / 10.1007 / s12272-018-1044-0; Two new naphthaleniclactone glycosides from Cassia obtusifolia L.seeds. Summary of the invention

[0018] The purpose of the present specification is to provide a naphthopyrone derivative of the following Chemical Formula 1, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

[0019] [Chemical formula 1]

[0020]

[0021] Another object of the present specification is to provide a composition which shows the effect of protecting nerve cells from damage caused by oxidative stress or inhibiting nerve cell death.

[0022] Another object of the present specification is to provide a composition which exhibits the effect of preventing or treating a neurological injury disease induced by damage or death of retinal nerve cells or hippocampal nerve cells.

[0023] Another object of the present invention is to provide a composition that exhibits an antioxidant effect.

[0024] In order to achieve the above object, one aspect of the present invention provides a naphthopyrone derivative of the following Chemical Formula 1, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

[0025] [Chemical formula 1]

[0026]

[0027] In another aspect, the present invention provides a method for producing one or more naphthopyrone derivatives selected from the following naphthopyrone derivatives of Chemical Formula 1 to Chemical Formula 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof.

[0028] [Chemical formula 1]

[0029]

[0030] [Chemical formula 2]

[0031]

[0032] [Chemical formula 3]

[0033]

[0034] [Chemical formula 4]

[0035]

[0036] [Chemical formula 5]

[0037]

[0038] On the other hand, the present invention provides a composition for protecting neurons from oxidative stress or for inhibiting neuronal cell death, comprising one or more naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same as an active ingredient, wherein the naphthopyrone derivative is one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5.

[0039] On the other hand, the present invention provides a composition for preventing or treating neurological damage diseases induced by damage or death of retinal neurons or hippocampal neurons, comprising one or more naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same as an active ingredient, wherein the naphthopyrone derivative is one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5.

[0040] On the other hand, the present invention provides an antioxidant composition comprising one or more naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same as an active ingredient, wherein the naphthopyrone derivative is one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5.

[0041] Effects of the Invention

[0042] The composition according to one aspect of the present invention has an antioxidant effect, and protects nerve cells from oxidative stress or inhibits the death of nerve cells, in particular, has the effect of inhibiting the damage or death of retinal nerve cells or hippocampal nerve cells caused by glutamate toxicity. Therefore, the composition according to one aspect of the present invention protects retinal nerve cells, and can therefore be used as a treatment or prevention of vision loss and decline and eye diseases associated with optic nerve damage, protects hippocampal nerve cells, and can therefore be used as a treatment or prevention of memory loss, learning loss, depression attacks and deterioration and degenerative brain nerve diseases caused by cranial nerve damage, and can be used to improve memory and learning ability, relieve mental health such as stress, and protect the optic nerve and improve eye health such as vision loss, and can therefore be used as a pharmaceutical composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1a and Figure 1b This is a graph showing the DPPH (α,α-diphenyl-β-picrylhydrazyl) radical scavenging activity of Cassia seed extract (ST) and Cassia seed bud extract (STS).

[0044] Figure 2a and Figure 2bThe figure shows the ABTS online antioxidant HPLC chromatogram of Cassia seed extract (ST) and Cassia seed bud extract (STS). The chromatogram of the blue line marked upward is a figure showing the components detected at ultraviolet (UV) 254nm, and the chromatogram of the red line marked downward is a figure showing the antioxidant components detected at UV / VIS (ultraviolet / visible light) 734nm after reacting with the ABTS reagent.

[0045] Figure 3a to Figure 3f The chromatograms of ABTS online antioxidant HPLC chromatograms of Cassia seed bud extracts (STS-C, STS-385, STS-465, STS-645 and STS-780) cultivated under various light conditions are shown. In each chromatogram (a) to (f), the chromatogram marked upward is a graph showing components detected at ultraviolet (UV) 254nm, and the chromatogram marked downward is a graph detecting antioxidant components that react with the ABTS reagent and are detected at UV / VIS (ultraviolet / visible light) 734nm. (a) The sample is an extract of Cassia seed bud cultivated for 2 weeks under light-shielding conditions (STS of Example 1), (b) the sample is an extract of Cassia seed bud cultivated for 2 weeks under normal light conditions (STS-C of Example 2), (c) the sample is an extract of Cassia seed bud cultivated for 2 weeks under 385nm LED illumination (STS-385 of Example 2), (d) the sample is an extract of Cassia seed bud cultivated for 2 weeks under 465nm LED illumination (STS-465 of Example 2), (e) the sample is an extract of Cassia seed bud cultivated for 2 weeks under 645nm LED illumination (STS-645 of Example 2), and (f) the sample is an extract of Cassia seed bud cultivated for 2 weeks under 780nm LED illumination (STS-780 of Example 2).

[0046] Figure 4 This is an HPLC chromatogram showing the peak of the main component separated from the Cassia seed sprout extract (STS) of Example 1 which is one embodiment of the present invention.

[0047] Figure 5a to Figure 5c The graph shows the protective effects of Cassia seed extract (ST) as Comparative Example 1 of the present invention, Cassia seed bud extract cultivated under light-shielding conditions (STS of Example 1), and Cassia seed bud extracts cultivated under various light conditions (STS, STS-C, STS-385, STS-465, STS-645, and STS-780 of Example 2) on retinal progenitor cells (R28) damaged by glutamate toxicity.

[0048] Figure 6a and Figure 6bIt is a graph showing the protective effect of each compound isolated from Cassia seed sprout extract (STS) (Compounds 1 to 5 of Examples 4 and 5, and Compounds A to X of Comparative Example 2) on retinal progenitor cell (R28) damage induced by glutamate toxicity according to the concentration.

[0049] Figures 7a to 7c The graph shows the protective effects of Cassia seed extract (ST) as Comparative Example 1 of the present invention, Cassia seed bud extract cultivated under light-shielding conditions (STS of Example 1), and Cassia seed bud extracts cultivated under various light conditions (STS, STS-C, STS-385, STS-465, STS-645, and STS-780 of Example 2) on hippocampal neurons (HT-22) damaged by glutamate toxicity.

[0050] Figure 8a and Figure 8b It is a graph showing the protective effect of each compound isolated from Cassia seed sprout extract (STS) (Compounds 1 to 5 of Examples 4 and 5, and Compounds A to X of Comparative Example 2) on hippocampal neuronal (HT-22) cells damaged by glutamate toxicity according to the concentration. DETAILED DESCRIPTION

[0051] One aspect of the present invention may relate to a naphthopyrone derivative of the following Chemical Formula 1, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof.

[0052] [Chemical formula 1]

[0053]

[0054] The naphthopyrone derivative of the above Chemical Formula 1 may be 7-hydroxymusizinyl-rubrofusarin-8'-O-glucopyranoside.

[0055] In this specification, "pharmaceutically acceptable" means that it can be used in animals, more specifically, in humans, by avoiding most toxic effects when used in general medicinal dosages, and can be recognized or recognized by the government or its approved regulatory agency, or listed in the pharmacopoeia, or recorded and understood in other general pharmacopoeias.

[0056] In the present specification, "pharmaceutically acceptable salt" refers to a salt according to one aspect of the present invention that is pharmaceutically acceptable and has the preferred pharmacological activity of the parent compound. The above salts may include: (1) acid addition salts formed from inorganic salts such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2,2,2]-oct-2-ene-1-carboxylic acid, glucoheptanoic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, or the like. or (2) a salt formed when an acid proton present in the parent compound is replaced.

[0057] In particular, in the present specification, "isomer" includes not only optical isomers (e.g., essentially pure enantiomers, essentially pure diastereomers or mixtures thereof), but also conformation isomers (i.e., isomers that differ only in the angle of one or more chemical bonds), position isomers (in particular, tautomers) or geometric isomers (e.g., cis-trans isomers).

[0058] In the present specification, "essentially pure" means, for example, when used in connection with enantiomers or diastereomers, that a specific compound exemplified as an enantiomer or diastereomer is present at about 90% or more, preferably at about 95% or more, more preferably at about 97% or more or at about 98% or more, further preferably at about 99% or more, and still further preferably at about 99.5% or more (w / w).

[0059] In this specification, "hydrate" refers to a compound to which water is bound, and is a broad concept including inclusion compounds in which there is no chemical bond between water and the compound.

[0060] In this specification, "solvate" refers to a higher order compound formed between molecules or ions of a solute and molecules or ions of a solvent.

[0061] One aspect of the present invention may relate to a method for producing a naphthopyrone derivative, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof, which is a method for producing one or more naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof selected from the following naphthopyrone derivatives of Chemical Formula 1 to Chemical Formula 5, comprising a process of isolating one or more naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof selected from the above naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof from Cassia seed (Cassia obtusifolia L. or Cassia tora L.) sprout.

[0062] [Chemical formula 1]

[0063]

[0064] [Chemical formula 2]

[0065]

[0066] [Chemical formula 3]

[0067]

[0068] [Chemical formula 4]

[0069]

[0070] [Chemical formula 5]

[0071]

[0072] The compound of the above chemical formula 1 (naphthopyrone derivative) may be 7-hydroxysomyosin-rubrofusarin-8'-O-pyranoglucoside, the compound of the above chemical formula 2 (naphthopyrone derivative) may be isotoralactone, the compound of the above chemical formula 3 (naphthopyrone derivative) may be toralactone, the compound of the above chemical formula 4 (naphthopyrone derivative) may be torosachrysone, and the compound of the above chemical formula 5 (naphthopyrone derivative) may be rubrofusarin.

[0073] The sprouts may be sprouts grown in a short period of time by germinating seeds, and may be sprouts grown by germinating seeds indoors, or may be sprouts generally sold.

[0074] The Cassia seed sprout or Cassia seed sprout plant of the above-mentioned production method can be a sprout or young plant that germinates from seeds of Cassia seed (Cassia obtusifolia L.) or Longgang Nancha (Cassia tora L. or Sennatora) and grows or cultivates naturally for 2 to 31 days. Specifically, it can be a sprout or young plant that grows or cultivates naturally for more than 2 days, more than 3 days, more than 4 days, more than 5 days, more than 6 days, more than 7 days, more than 8 days, more than 9 days, more than 10 days, more than 11 days, more than 12 days, more than 13 days, more than 14 days, more than 15 days, more than 16 days, more than 17 days, more than 18 days, more than 19 days, more than 20 days, more than 21 days, more than 22 days, more than 23 days, more than 24 days, more than 25 days, more than 26 days, more than 27 days, more than 28 days, more than 29 days or more than 30 days. The buds or young plants may be buds or young plants that have been grown naturally or cultivated for less than 31 days, less than 30 days, less than 29 days, less than 28 days, less than 27 days, less than 26 days, less than 25 days, less than 24 days, less than 23 days, less than 22 days, less than 21 days, less than 20 days, less than 19 days, less than 18 days, less than 17 days, less than 16 days, less than 15 days, less than 14 days, less than 13 days, less than 12 days, less than 11 days, less than 10 days, less than 9 days, less than 8 days, less than 7 days, less than 6 days, less than 5 days, less than 4 days, or less than 3 days. More specifically, they may be buds or young plants that have been grown naturally or cultivated for a period selected from a period of 2 to 21 days.

[0075] The above-mentioned Cassia seed (Cassia obtusifolia L. or Cassia tora L.) refers to a herbaceous plant of the order Rosales, family Leguminosae, in the class Dicotyledons of the Angiosperms. Cassia seed is the mature seed of the above-mentioned Cassia seed (Cassia obtusifolia L.) or Longgang Nancha (Cassia tora L. or Sennatora). Although the Cassia seed extract is known to have the effects of improving diabetes, improving dyslipidemia, protecting the liver, antibacterial, and lowering blood pressure (Dong, X. et al. Mol. Med. Rep. 2017, 16, 2331-2346), it is not known to improve the damage or inhibit death of retinal neurons or hippocampal neurons caused by oxidative stress or drug toxicity, especially glutamate.

[0076] The Cassia seed bud or young plant can be the whole or part of the Cassia seed bud or young plant. The part can be the front tip or the above-ground part or the underground part. The above-ground part can be a stem, a leaf, a flower or a combination thereof. The underground part can be a root. The plant can grow naturally or be cultivated artificially. The Cassia seed bud of the present invention can be easily cultivated indoors and can be cultivated and used in a short period of several weeks without providing additional nutrients, so it has the advantage of greater industrial applicability.

[0077] The Cassia seed bud extract includes a crude extract or further includes a fraction obtained by further fractionation of the extract. Specifically, the Cassia seed bud extract can be a crude extract, a fraction, or a combination thereof of the Cassia seed bud. The crude extract refers to a substance obtained by contacting the Cassia seed bud with an extraction solvent. The fraction refers to a substance obtained by separating a substance containing a specific component from the crude extract. The extract or its fraction can be included in the composition of the present invention in the form of an extract of the Cassia seed bud plant, its fraction, or a sub-fraction, each or a mixture thereof. The sub-fraction can be obtained by an ultrafiltration membrane with a cutoff value, and can be obtained by column chromatography or solvent fractionation. The Cassia seed bud extract or fraction can contain one or more naphthopyrone derivatives selected from the naphthopyrone derivatives of the above-mentioned chemical formulas 1 to 5.

[0078] The separation can be performed by filtration, precipitation, centrifugation, solvent fractionation, chromatography or a combination thereof. The chromatography is prepared for separation based on various conditions, i.e., size, charge, hydrophobicity or affinity, and can be ion exchange chromatography, affinity chromatography, size exclusion chromatography, HPLC, high performance liquid chromatography, column chromatography, reversed phase column chromatography or a combination thereof.

[0079] The extraction may include incubating the plant in a solvent for a certain period of time. The extraction may be performed with or without stirring, or may include heating. The incubation may be performed with or without stirring at room temperature or at reflux temperature. The incubation temperature may be appropriately selected depending on the selected solvent. For example, it may be room temperature to reflux temperature, 30°C to reflux temperature, 40°C to reflux temperature. The heating may include heating to 50°C, 60°C, 70°C, 80°C, or reflux temperature. The heating may be heating to 50°C to reflux temperature, 60°C to reflux temperature, 70°C to reflux temperature, 80°C to reflux temperature, or reflux temperature. The extraction time can be changed according to the selected temperature, and can be 1 hour to 2 months, for example, it can be 1 hour to 1 month, 1 hour to 15 days, 1 hour to 10 days, 1 hour to 5 days, 1 hour to 3 days, 1 hour to 2 days, 1 hour to 1 day, 5 hours to 1 month, 5 hours to 15 days, 5 hours to 10 days, 5 hours to 5 days, 5 hours to 3 days, 5 hours to 2 days, 5 hours to 1 day, 10 hours to 1 month, 10 hours to 15 days, 10 hours to 10 days, 10 hours to 5 days, 10 hours to 3 days, or 10 hours to 2 days. The extraction can be reflux extraction of the plant in a solvent. The solvent can have a volume of 1, 2, 5, 10 or 15 times or more relative to the weight of the plant. The solvent can have a volume of 1 to 15 times, 2 to 15 times, 5 to 15 times, 10 to 15 times or about 15 times relative to the weight of the plant. The plants may be plants dried in a shade or shading device or in a warm wind or drying device.

[0080] As the above-mentioned extraction method, common methods in this field such as filtration, hot water extraction, precipitation extraction, cold precipitation, microwave extraction, reflux cooling extraction, pressure extraction, subcritical extraction, supercritical extraction and ultrasonic extraction can be used. For example, it can be a precipitation extraction method. The precipitation extraction can be warm precipitation or precipitation at room temperature, and can be extracted 1 to 5 times. The above-mentioned Cassia seed bud plant can be contacted with 0.1 to 10 times or 1 to 6 times the extraction solvent. The cold precipitation extraction temperature can be 20°C to 40°C. The warm precipitation or heating extraction temperature can be 40°C to 100°C. The cold precipitation extraction time can be 24 hours to 120 hours, and the warm precipitation or heating extraction time can be 0.5 hours to 48 hours.

[0081] The above-mentioned extraction may also include removing the solvent from the obtained extract by known methods such as evaporation or reduced pressure concentration. The above-mentioned extraction may also include drying the obtained extract by freeze drying to produce a dry extract. The above-mentioned reduced pressure concentration may utilize a vacuum reduced pressure concentrator or a vacuum rotary evaporator. In addition, the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying.

[0082] The manufacturing method may further include extracting the Cassia seed buds with a solvent selected from water, C1 to C6 alcohols and mixed solvents thereof. The alcohol may be a C1 to C3 alcohol, a C1 to C4 alcohol, a C1 to C5 alcohol or a C1 to C6 alcohol. The alcohol may be a primary alcohol. The C1 to C6 alcohol may be methanol, ethanol, propanol, isopropanol, butanol or a mixture thereof.

[0083] In addition, the above-mentioned production method further includes a process of fractionating with one or more selected from water, ethyl acetate, hexane, dichloromethane, chloroform, methanol, ethanol, acetone, and a mixed solvent thereof. The above-mentioned production method also includes a process of producing a Cassia seed bud fraction containing 1 to 20 weight % of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, based on the total weight of the extract obtained by the above-mentioned process of extracting the Cassia seed bud. Specifically, it can include the process of producing a Cassia seed bud fraction containing 1 weight % or more, 2 weight % or more, 3 weight % or more, 4 weight % or more, 5 weight % or more, 6 weight % or more, 7 weight % or more, 8 weight % or more, 9 weight % or more, 10 weight % or more, 11 weight % or more, 12 weight % or more, The process of producing a Cassia seed bud fraction containing 13% or more, 14% or more, 15% or more, 16% or more, 17% or more, 18% or more or 19% or more of the Cassia seed bud fraction may include the process of producing a Cassia seed bud fraction containing 20% ​​or less, 19% or less, 18% or less, 17% or less, 16% or less, 15% or less, 14% or less, 13% or less, 12% or less, 11% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4% or less, 3% or less or 2% or less of the Cassia seed bud fraction.

[0084] The naphthopyrone derivatives, stereoisomers, pharmaceutically acceptable salts, hydrates or solvates thereof selected from the naphthopyrone derivatives of the above Chemical Formulas 1 to 5 may be contained in an amount of 1 to 50% by weight, based on the total weight of the Cassia seed bud extract or fraction. Specifically, the amount may be 1% by weight or more, 2% by weight or more, 3% by weight or more, 4% by weight or more, 5% by weight or more, 6% by weight or more, 7% by weight or more, 8% by weight or more, 9% by weight or more, 10% by weight or more, 11% by weight or more, 12% by weight or more, 13% by weight or more, 14% by weight or more, 15% by weight or more, 16% by weight or more, 17% by weight or more, 18% by weight or more, 19% by weight or more, 20% by weight or more, 21% by weight or more, 22% by weight or more, 23% by weight or more, 24% by weight or more, 25% by weight or more, 26% by weight or more, 27% by weight or more, 28% by weight or more, 29% by weight or more, 30% by weight or more, 31% by weight or more, 32% by weight or more, 33% by weight or more, 34% by weight or more, 35% by weight or more, 36% by weight or more, 37% by weight or more, 38% by weight or more, 39% by weight or more, 40% by weight or more, 41% by weight or more, 42% by weight or more, 43% by weight or more, 44% by weight or more, 45% by weight or more, 46% by weight or more, 47% by weight or more, 48% by weight 6 weight % or more, 17 weight % or more, 18 weight % or more, 19 weight % or more, 20 weight % or more, 21 weight % or more, 22 weight % or more, 23 weight % or more, 24 weight % or more, 25 weight % or more, 26 weight % or more, 27 weight % or more, 28 weight % or more, 29 weight % or more, 30 weight % or more, 31 weight % or more, 32 weight % or more, 33 weight % or more, 34 weight % or more, 35 weight % or more, 36 weight % or more, 37 weight % or more, 38 weight % or more, 39 weight % or more, 40 weight % or more, 41 weight % or more, 42 weight % or more, 43 weight % or more %, 44 wt % or more, 45 wt % or more, 46 wt % or more, 47 wt % or more, 48 wt % or more, or 49 wt % or more, and may include 50 wt % or less, 49 wt % or less, 48 ​​wt % or less, 47 wt % or less, 46 wt % or less, 45 wt % or less, 44 wt % or less, 43 wt % or less, 42 wt % or less, 41 wt % or less, 40 wt % or less, 39 wt % or less, 38 wt % or less, 37 wt % or less, 36 wt % or less, 35 wt % or less, 34 wt % or less, 33 wt % or less, 32 wt % or less, 31 wt % or less, 30 wt % or less, %, 29 weight % or less, 28 weight % or less, 27 weight % or less, 26 weight % or less, 25 weight % or less, 24 weight % or less, 23 weight % or less, 22 weight % or less, 21 weight % or less, 20 weight % or less, 19 weight % or less, 18 weight % or less, 17 weight % or less, 16 weight % or less, 15 weight % or less, 14 weight % or less, 13 weight % or less, 12 weight % or less, 11 weight % or less, 10 weight % or less, 9 weight % or less, 8 weight % or less, 7 weight % or less, 6 weight % or less, 5 weight % or less, 4 weight % or less, 3 weight % or less, or 2 weight % or less.

[0085] One aspect of the present invention may relate to a composition for protecting nerve cells from oxidative stress or for inhibiting nerve cell death, comprising one or more selected from the group consisting of naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtusifolia L. or Cassia tora L.) bud extract containing the same, or a Cassia seed bud fraction containing the same as an active ingredient, wherein the naphthopyrone derivative is one or more selected from the naphthopyrone derivatives of the above Chemical Formulas 1 to 5. The above one or more naphthopyrone derivatives selected from the above Chemical Formulas 1 to 5 may be naphthopyrone derivatives selected from the above Chemical Formula 1 and one or more naphthopyrone derivatives selected from the above Chemical Formulas 2 to 5. The descriptions regarding the naphthopyrone derivatives, pharmaceutically acceptable salts, stereoisomers, hydrates, solvates, Cassia seed buds, Cassia seed bud extracts or fractions of the above Chemical Formulae 1 to 5 are as described above.

[0086] The above-mentioned composition can be a composition for protecting nerve cells or for inhibiting nerve cell death, specifically, it can be a composition for protecting nerve cells from oxidative stress induced by metabolic toxicity, neurotoxicity, chemical causes, etc. or for inhibiting nerve cell death. The above-mentioned oxidative stress can be induced by glutamate, glutamate toxicity, or glutamate neurotoxicity, or it can be oxidative stress caused by calcium homeostasis disorder, mitochondrial dysfunction, excitotoxicity, nutrient factor depletion, etc. similar to the oxidative stress induced by glutamate neurotoxicity. Specifically, the above-mentioned composition can reduce, inhibit, improve, or prevent the damage of retinal nerve cells or hippocampal nerve cells induced by glutamate, glutamate toxicity, or glutamate neurotoxicity, or resuscitate or regenerate dead retinal nerve cells or hippocampal nerve cells, and can be used as a defense mechanism for oxidative stress induced by glutamate neurotoxicity, calcium homeostasis disorder, mitochondrial dysfunction, excitotoxicity, nutrient factor depletion, etc. to protect nerve cells, or inhibit nerve cell death.

[0087] According to one embodiment of the present invention, the Cassia seed bud extract has a DPPH free radical scavenging effect that is more than 1.7 times better than that of the Cassia seed extract (Test Example 1), and the content of the compounds of the above chemical formulas 1 to 5 (naphthopyrone derivatives) equivalent to antioxidant components is high, thereby showing a more excellent antioxidant effect (Test Example 2). The antioxidant effect of the composition of the present invention as a defense mechanism against oxidative stress is activated, thereby confirming that it has an excellent effect in protecting neurons or inhibiting neuronal cell death.

[0088] The composition can be treated before, at the same time as, or after nerve cell damage or death occurs. The composition can contain 0.001 wt % to 80 wt % of one or more selected from naphthopyrone derivatives, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, Cassia seed (Cassia obtuse or Cassia small) bud extracts containing the same, or Cassia seed bud fractions containing the same, relative to the total weight of the composition. The naphthopyrone derivative is one or more selected from the naphthopyrone derivatives of Chemical Formulas 1 to 5. Specifically, it can contain 0.001 wt %, 0.01 wt %, or more. The amount of the organic compound may be greater than 0.05 wt%, greater than 0.1 wt%, greater than 1 wt%, greater than 2 wt%, greater than 3 wt%, greater than 4 wt%, greater than 5 wt%, greater than 10 wt%, greater than 20 wt%, greater than 30 wt%, greater than 40 wt% or greater than 60 wt%, and may include less than 80 wt%, less than 60 wt%, less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, less than 4 wt%, less than 3 wt%, less than 2 wt% or less than 1 wt%.

[0089] In addition, the Cassia seed bud extract or fraction contains 1 to 50 weight % of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, based on the total weight of the Cassia seed bud extract or fraction. Specifically, it may contain 1 weight % or more, 2 weight % or more, 3 weight % or more, 4 weight % or more, 5 weight % or more, 6 weight % or more, 7 weight % or more, 8 weight % or more, 9 weight % or more, 10 weight % or more, 11 weight % or more, 12 weight % or more, 13 weight % or more, 14 weight % or more %, 15% by weight or more, 16% by weight or more, 17% by weight or more, 18% by weight or more, 19% by weight or more, 20% by weight or more, 21% by weight or more, 22% by weight or more, 23% by weight or more, 24% by weight or more, 25% by weight or more, 26% by weight or more, 27% by weight or more, 28% by weight or more, 29% by weight or more, 30% by weight or more, 31% by weight or more, 32% by weight or more, 33% by weight or more, 34% by weight or more, 35% by weight or more, 36% by weight or more, 37% by weight or more, 38% by weight or more, 39% by weight or more, 40% by weight or more, 41% by weight or more, 42% by weight or more %, 43% or more, 44% or more, 45% or more, 46% or more, 47% or more, 48% or more, or 49% or more, and may contain 50% or less, 49% or less, 48% or less, 47% or less, 46% or less, 45% or less, 44% or less, 43% or less, 42% or less, 41% or less, 40% or less, 39% or less, 38% or less, 37% or less, 36% or less, 35% or less, 34% or less, 33% or less, 32% or less, 31% or less, or 32% or less. 0 weight % or less, 29 weight % or less, 28 weight % or less, 27 weight % or less, 26 weight % or less, 25 weight % or less, 24 weight % or less, 23 weight % or less, 22 weight % or less, 21 weight % or less, 20 weight % or less, 19 weight % or less, 18 weight % or less, 17 weight % or less, 16 weight % or less, 15 weight % or less, 14 weight % or less, 13 weight % or less, 12 weight % or less, 11 weight % or less, 10 weight % or less, 9 weight % or less, 8 weight % or less, 7 weight % or less, 6 weight % or less, 5 weight % or less, 4 weight % or less, 3 weight % or less, or 2 weight % or less.

[0090] The nerve cells may be central nerves, peripheral nerves or optic nerves associated with the hippocampus of the brain or the retina of the eye, and specifically, may be retinal nerve cells or hippocampal nerve cells.

[0091] The retinal nerve cells can be one or more cells selected from rod cells, cone cells, bipolar cells, retinal amacrine cells, horizontal cells and retinal ganglion cells. The rod cells and cone cells are nerve cells that sense light and recognize things, the bipolar cells, retinal amacrine cells and horizontal cells are nerve cells that transmit visual information to retinal ganglion cells, and the retinal ganglion cells are nerve cells that transmit visual information of the retina to the brain. The R28 retinal progenitor cells or R28 retinal neural progenitor cells of one embodiment of the present invention are cells that express the characteristics of various constituent cells of the retina and can survive even if they are moved to the retina. They can also be effectively used for cell death and cytotoxicity studies related to glutamate or GABA receptor expression associated with glaucoma for cell death caused by hypoxia or serum deficiency. In addition, R28 cells can differentiate from retinal progenitor cells into retinal ganglion cells according to culture conditions, and can be used not only for basic cytological research of the retina but also for basic research of retinal ganglion cells (Jungil Lee, Jaewoo Kim, Journal of the Korean Ophthalmological Association, 2009, 50, 919-922).

[0092] The hippocampal nerve cells are nerve cells of the hippocampus, which is a structure of the medial temporal lobe of the brain. The hippocampus is weak to physiological / oxidative stress stimulation, causing damage to cognitive function due to stress. It is known as a central tissue. The stress can cause behavioral changes related to the structure and brain cell generation of the hippocampus, synaptic plasticity and hippocampus (Kim Eun-joo, Korean Journal of Psychology: Cognition and Biology, 2012, 24, 65-88). The HT-22 hippocampal nerve cells of one embodiment of the present invention are sensitive to glutamate and can be used as an in vitro model system for studying neurotoxicity induced by oxidative stress (Liu, J. et al. Life Science, 2009, 84, 267-271; Kim Jihyun Kim, Soonsil Jeon, Korean Journal of Food and Nutrition Science, 2017, 46, 886-890).

[0093] According to one embodiment of the present invention, when R28 cells induced by oxidative stress by glutamate treatment were treated with Cassia seed bud extract and naphthopyrone derivatives of Chemical Formulas 1 to 5, respectively, the survival rate of cells reduced by glutamate increased to a level corresponding to that of the control group, and such retinal cell protection effect was superior to that of the Cassia seed extract as a comparative example, confirming that the composition of the present invention is excellent in protecting retinal nerve cell damage caused by oxidative stress or inhibiting retinal nerve cell death (Test Examples 3 and 4).

[0094] In addition, according to one embodiment of the present invention, when HT-22 cells induced by oxidative stress by glutamate treatment were treated with Cassia seed bud extract and naphthopyrone derivatives of the above Chemical Formulas 1 to 5, respectively, the survival rate of cells reduced by glutamate increased to a level corresponding to that of the control group, and such hippocampal neuron protective effect was superior to that of the Cassia seed extract used as a comparative example, confirming that the composition of the present invention is excellent in protecting hippocampal neuron damage caused by oxidative stress or inhibiting hippocampal neuron death (Test Examples 5 and 6).

[0095] One aspect of the present invention may relate to a composition for preventing or treating a neurological injury disease induced by damage or death of retinal nerve cells or hippocampal nerve cells, which comprises one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned chemical formula 1 to chemical formula 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, Cassia seed (Cassia obtuse or Cassia small) bud extracts containing the same, or Cassia seed bud fractions containing the same as active ingredients, or may relate to a composition for preventing or treating a neurological injury disease induced by glutamate neurotoxicity. One or more naphthopyrone derivatives selected from the above-mentioned chemical formula 1 to chemical formula 5 may be naphthopyrone derivatives selected from the above-mentioned chemical formula 1, and one or more naphthopyrone derivatives selected from the above-mentioned chemical formula 2 to chemical formula 5. The descriptions regarding the naphthopyrone derivatives, pharmaceutically acceptable salts, stereoisomers, hydrates, solvates, Cassia seed buds, Cassia seed bud extracts or fractions, and contents of the above Chemical Formulas 1 to 5 are as described above.

[0096] The above-mentioned neurological injury disease induced by the damage or death of retinal nerve cells or hippocampal nerve cells can be caused by glutamate neurotoxicity, and the description of the above-mentioned retinal nerve cells and hippocampal nerve cells is as described above. The above-mentioned neurological injury disease can be selected from diabetic retinopathy caused by retinal nerve cell damage or death, macular degeneration, visual impairment caused by retinal cell damage, retinitis pigmentosa, retinal detachment, retinal vascular occlusion and glaucoma. It can be selected from memory loss, learning impairment, depression, amyotrophic lateral sclerosis (Lou Gehrig's disease), Parkinson's disease and cerebral ischemic nerve damage caused by hippocampal nerve cell damage or death. One or more, but as long as it is a disease caused by the damage or death of retinal nerve cells or hippocampal nerve cells induced by glutamate neurotoxicity, there is no limitation.

[0097] The above-mentioned glutamate neurotoxicity refers to the toxicity exhibited by glutamate acting as an oxidative neurotoxic substance due to a sharp increase in the glutamate concentration outside the nerve cells. The above-mentioned glutamate neurotoxicity may be induced by glutamate flowing in from outside the individual, and the above-mentioned influx of glutamate may be caused by the intake or administration of glutamate-containing foods, glutamate-containing therapeutic agents, glutamate-containing preventive agents, glutamate-containing antibiotics, etc., but is not limited thereto.

[0098] One or more naphthopyrone derivatives selected from the naphthopyrone derivatives of the above Chemical Formulas 1 to 5 can be isolated or purified from Cassia seed (Cassia obtuse or Cassia small) buds, specifically, can be isolated or purified from Cassia seed bud ethanol extract, and can be a derivative obtained by fractionating the Cassia seed bud ethanol extract with ethyl acetate, separating the ethyl acetate fraction by chromatography, or fractionating the ethyl acetate fraction again with a mixed solvent of hexane, dichloromethane and methanol, and then subjecting the fraction to chromatography.

[0099] The Cassia seed bud extract may be an extract obtained by extracting the Cassia seed bud with a solvent selected from water, C1 to C6 alcohols and mixed solvents thereof. The alcohol may be a C1 to C3 alcohol, a C1 to C4 alcohol, a C1 to C5 alcohol or a C1 to C6 alcohol. The alcohol may be a primary alcohol. The C1 to C6 alcohol may be methanol, ethanol, propanol, isopropanol, butanol or a mixture thereof.

[0100] The above-mentioned fractionation may further include a process of fractionating with one or more solvents selected from water, ethyl acetate, hexane, dichloromethane, chloroform, methanol, ethanol, acetone, and a mixed solvent thereof.

[0101] According to one embodiment of the present invention, one or more naphthopyrone derivatives selected from the naphthopyrone derivatives of the above-mentioned Chemical Formulas 1 to 5 are used as antioxidant ingredients. Compared with the Cassia seed extract, the Cassia seed bud extract contains a higher content (Test Example 2). The naphthopyrone derivatives are excellent in protecting retinal neurons (R28) and hippocampal neurons (HT-22) that are damaged or even dead by glutamate treatment (Test Examples 4 and 6). It is confirmed that the naphthopyrone derivatives of the above-mentioned Chemical Formulas 1 to 5 are effective ingredients of Cassia seed buds that have the effect of preventing or treating neurological damage diseases induced by glutamate neurotoxicity.

[0102] One aspect of the present invention may relate to an antioxidant composition, which contains one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned chemical formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, Cassia seed (Cassia obtuse or Cassia small) bud extracts containing the same, or Cassia seed bud fractions containing the same as active ingredients. One or more naphthopyrone derivatives selected from the above-mentioned chemical formulas 1 to 5 may be naphthopyrone derivatives selected from the above-mentioned chemical formula 1, and one or more naphthopyrone derivatives selected from the above-mentioned chemical formulas 2 to 5. The description of the naphthopyrone derivatives, pharmaceutically acceptable salts, isomers, hydrates, solvates, Cassia seed buds, Cassia seed bud extracts or fractions, and contents of the above-mentioned chemical formulas 1 to 5 is as described above.

[0103] In another aspect of the present invention, it may involve a method for protecting nerve cells from oxidative stress or inhibiting nerve cell death, comprising administering to an individual who needs to protect nerve cells from oxidative stress or inhibit nerve cell death one or more of the naphthopyrone derivatives selected from the above-mentioned chemical formula 1 to chemical formula 5, its stereoisomers, its pharmaceutically acceptable salts, its hydrates, or its solvates, a Cassia seed (Cassia obtuse or Cassia small) bud extract containing the same, or a Cassia seed bud fraction containing the same, to protect nerve cells from oxidative stress or inhibit nerve cell death. In one aspect of the present invention, the administration of the above method can be implemented according to the administration method and administration dosage described in this specification.

[0104] In another aspect of the present invention, it may involve a method for preventing or treating a neurological injury disease induced by damage or death of retinal neurons or hippocampal neurons, including administering to an individual who needs to prevent or treat a neurological injury disease induced by damage or death of retinal neurons or hippocampal neurons, one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned chemical formula 1 to chemical formula 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse or Cassia small) bud extract containing the same, or a Cassia seed bud fraction containing the same, to prevent or treat a neurological injury disease induced by damage or death of retinal neurons or hippocampal neurons. In one aspect of the present invention, the administration of the above method can be implemented according to the administration method and administration dosage described in this specification.

[0105] In another aspect of the present invention, it may involve an antioxidant method comprising administering to an individual in need of antioxidant activity one or more naphthopyrone derivatives selected from the naphthopyrone derivatives of the above Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse or Cassia small) bud extract containing the same, or a Cassia seed bud fraction containing the same. In one aspect of the present invention, the administration of the above method may be implemented according to the administration method and administration dosage described in this specification.

[0106] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of a pharmaceutical composition for protecting neurons from oxidative stress or inhibiting neuronal cell death.

[0107] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of a pharmaceutical composition for preventing or treating neurological damage diseases induced by damage or death of retinal neurons or hippocampal neurons.

[0108] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, a Cassia seed (Cassia obtuse or Cassia small) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of an antioxidant pharmaceutical composition.

[0109] In another aspect of the present invention, the present invention may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of a cosmetic composition for protecting neurons from oxidative stress or inhibiting neuronal cell death.

[0110] In another aspect of the present invention, the present invention may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of a cosmetic composition for improving, preventing or treating neurological damage diseases induced by damage or death of retinal neurons or hippocampal neurons.

[0111] In another aspect of the present invention, the present invention may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of an antioxidant cosmetic composition.

[0112] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, a Cassia seed (Cassia obtuse or Cassia small) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of a quasi-drug composition for protecting neurons from oxidative stress or inhibiting neuronal cell death.

[0113] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of a quasi-drug composition for improving, preventing or treating a neurological injury disease induced by damage or death of retinal neurons or hippocampal neurons.

[0114] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, a Cassia seed (Cassia obtuse or Cassia small) bud extract containing the same, or a Cassia seed bud fraction containing the same, for the manufacture of an antioxidant quasi-drug composition.

[0115] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same for protecting neurons from oxidative stress or inhibiting neuronal cell death.

[0116] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same for preventing or treating neurological damage diseases induced by damage or death of retinal neurons or hippocampal neurons.

[0117] In another aspect of the present invention, it may involve the antioxidant use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, stereoisomers thereof, pharmaceutically acceptable salts thereof, hydrates thereof, or solvates thereof, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same.

[0118] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for protecting neurons from oxidative stress or inhibiting the death of neurons.

[0119] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same, for preventing or treating neurological damage diseases induced by damage or death of retinal neurons or hippocampal neurons.

[0120] In another aspect of the present invention, it may involve the use of one or more naphthopyrone derivatives selected from the group consisting of one or more naphthopyrone derivatives selected from the above-mentioned Chemical Formulas 1 to 5, their stereoisomers, their pharmaceutically acceptable salts, their hydrates, or their solvates for antioxidant purposes, a Cassia seed (Cassia obtuse leaf or Cassia small leaf) bud extract containing the same, or a Cassia seed bud fraction containing the same.

[0121] The composition of the present invention may be a pharmaceutical composition or a cosmetic composition.

[0122] The pharmaceutical composition according to one aspect of the present invention can be formulated into oral or non-oral administration. In terms of formulation, commonly used fillers, extenders, adhesives, wetting agents, disintegrants, surfactants and other diluents or excipients can be used to prepare. Solid preparations for oral administration include tablets, pills, powders, granules, soft capsules or hard capsules, etc., and such solid preparations are prepared by mixing at least one or more excipients in more than one compound, for example, starch, calcium carbonate, sucrose (sucrose) or lactose (lactose), gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talcum powder are also used. As liquid preparations for oral administration, equivalent to suspensions, liquid solutions, emulsions, syrups, etc., in addition to water and liquid paraffin as simple diluents that are often used, various excipients can also be included, for example, wetting agents, sweeteners, flavoring agents, preservatives, etc. Preparations for non-oral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As non-aqueous solvents and suspending solvents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, etc. can be used. As the base of the suppository, synthetic fatty acid esters (witepsol), polyethylene glycol, Tween 61, cocoa butter, lauric acid esters, glycerin gelatin, etc. can be used.

[0123] The pharmaceutical administration of the composition according to one aspect of the present invention can also be used in the form of their pharmaceutically acceptable salts, and can be used alone or in combination with other pharmaceutically active compounds, and in the form of appropriate combinations. As the above-mentioned salt, as long as it is a pharmaceutically acceptable salt, there is no particular limitation, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, etc. can be used. In addition, the non-oral administration dosage form can be a skin patch, an ointment, a cream, an eye drop, a spray or an injection.

[0124] In another aspect, the present invention may be a method for protecting neural cells in an individual, comprising the step of administering the above-mentioned pharmaceutical composition to the individual.

[0125] The above-mentioned individual can be a mammal, for example, a human, a cow, a horse, a pig, a dog, a sheep, a goat or a cat. The above-mentioned mammal can be a human. The effective administration amount of the compound of the present invention for the human body can vary according to the patient's age, weight, sex, administration method, health status and disease severity.

[0126] The above-mentioned administration can be carried out in various dosage forms for oral administration or non-oral administration such as intravenous, transdermal, subcutaneous, epithelial or intramuscular administration. For formulation, it can be produced using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc.

[0127] The administration can be administered by methods known in the art. The administration can be appropriately administered to an individual by any method, for example, intravenous, intramuscular, oral, or subcutaneous administration. The administration can be administered systemically or locally. The administration can be administered minimally to a site where nerve cells are expected to be present or generated.

[0128] The composition of the present invention may be a quasi-drug composition.

[0129] The above-mentioned "quasi-drugs" are drugs that exclude articles used as drugs according to the Pharmaceutical Affairs Law, and refer to drugs based on the classification standards separately stipulated by the Ministry of Health and Welfare for articles that have a milder effect on the human body than pharmaceuticals used for the treatment or prevention of diseases. Therefore, it can include fibers and rubber products used to treat or prevent diseases in humans or animals; instruments or non-instrumental articles and their similarities that have a mild or no direct effect on the human body; or bactericides and insecticides used to block infectious diseases. The above-mentioned quasi-drug composition of the present invention can be used to improve, protect or treat nerve cells from damage by oxidative stress, and can be used to prevent or improve nerve damage diseases induced by glutamate neurotoxicity.

[0130] The above-mentioned quasi-drug composition may further include an excipient or carrier acceptable as a quasi-drug.

[0131] The above-mentioned quasi-drug composition can be prepared in the form of a skin spread, cream, ointment, eye drop, or spray.

[0132] The present invention will be described in more detail below by way of examples and test examples. However, these examples and test examples are provided for illustrative purposes only and the scope of the present invention is not limited to these examples and test examples.

[0133] [Example 1] Preparation of Cassia Seed Bud Extract

[0134] Unsterilized domestic Cassia seeds (Cassia obtusifolia L.) (Handsherb, January 2016) were purchased, sterilized with bleaching powder (CaOCl2) at a concentration of 20 mg / mL for 15 minutes, washed thoroughly with water, soaked in pure water for 4 hours, and cultivated under light-shielding conditions for 6 days. The Cassia seed buds obtained by the above cultivation were dried at 32°C hot air (38°C) for 22 days, crushed to obtain 698.4g of dried plants. Then, the above dried plants were placed in an extraction container, 5.5L of ethanol was added, mixed and stirred at room temperature, extracted for 7 days, and the above mixture was gravity filtered using Whatman filter paper with a film thickness of 0.34mm and a glass funnel with a diameter of 30cm. This process was repeated twice ("extraction-filtration" twice) to obtain a filtered extract. The filtered extract was placed in a vacuum concentrator and concentrated by completely evaporating the solvent at 35°C under reduced pressure to obtain 66.7 g of Cassia seed bud extract (hereinafter referred to as "STS") (yield 9.55%).

[0135] [Comparative Example 1] Preparation of Cassia Seed Extract

[0136] Unsterilized domestic Cassia seeds (Cassia obtusifolia L.) (Handsherb, January 2016) were purchased, crushed, placed in a 15 mL extraction container, 11 mL of ethanol was added, mixed and stirred at room temperature, extracted for 7 days, and the mixture was subjected to gravity filtration ("extraction-filtration" 2 times) using Whatman filter paper with a film thickness of 0.34 mm and a glass funnel with a diameter of 10 cm to obtain a filtered extract (hereinafter referred to as "ST"). The filtered extract was placed in a vacuum concentrator, and the solvent was completely evaporated and concentrated at 35°C under reduced pressure. (Yield 3.34%).

[0137] [Example 2] Production of Cassia seed bud extract cultivated under various light conditions

[0138] In order to compare and analyze the components of the Cassia seed bud extract (STS) cultivated under the shading condition of the above Example 1 and the components of the Cassia seed bud extract cultivated under the specific light condition, the Cassia seed bud was cultivated by making the light wavelength condition different. The Cassia seed bud was cultivated by the same method as in the above Example 1, and the light condition when cultivating the Cassia seed bud was not the shading condition, and the light of the selective light quality of the large wavelengths of the fluorescent lamp, LED 385nm, 465nm, 645nm, and 780nm was irradiated, and each was cultivated for 6 days. Using the five types of buds obtained by the above-mentioned cultivation, respective Cassia seed bud extracts were produced by the same method as in the above-mentioned Example 1. The extract obtained by cultivation under fluorescent lamp conditions was called "STS-C", the extract obtained by cultivation under LED 385nm light conditions was called "STS-385", the extract obtained by cultivation under LED 465nm light conditions was called "STS-465", the extract obtained by cultivation under LED 645nm light conditions was called "STS-645", and the extract obtained by cultivation under LED 780nm light conditions was called "STS-780".

[0139] [Example 3] Preparation of Cassia Seed Bud Extract (STS) Fractions

[0140] After 66.7 g of the Cassiae bud extract (STS) of Example 1 was suspended in 600 mL of water, 1.8 L of ethyl acetate was added, and the mixture was shaken and stirred at room temperature. The mixture was fractionated four times every 2 hours to obtain 23.5 g of ethyl acetate fraction (STS-EA). The ethyl acetate fraction was fractionated using dichloromethane (or ethyl acetate) alone or 2.4 L of a mixed solvent of n-hexane, dichloromethane (or ethyl acetate), and methanol (or ethanol) (the mixing ratio (volume ratio) of n-hexane:dichloromethane was 1:1, and the mixing ratio (volume ratio) of dichloromethane:methanol was 200:1, 50:1, 10:1, 5:1, 3:1, and 1:1) as a fractionation solvent, thereby producing the above-mentioned STS-EA fraction, i.e., the Cassiae bud extract (STS) fraction.

[0141] Specifically, the ethyl acetate fraction (STS-EA) was brought into contact with the eight fractionation solvents, stirred in celite, the solvent was evaporated, and developed using a silica resin packed in a column having a diameter of 10 cm and a length of 20 cm. Chromatography was performed using the solvents to obtain 20 fractions (STS-EA-fraction 1 to STS-EA-fraction 20).

[0142] [Example 4] Isolation of Compound 1 from Cassia Seed Bud Extract

[0143] After the STS-EA-fraction 12 produced in the above Example 3 (the elution solvent in Example 3 is the second of the three fractions in which the mixing ratio of dichloromethane (or ethyl acetate): methanol (or ethanol) is 10:1) was concentrated under reduced pressure, chromatography for separation was performed under the conditions of the following separation method 1, and compound 1 (14.0 mg, 0.002% of the dry plant weight; 0.021% of the dry extract weight) having the structure of the above chemical formula 1 was separated under the conditions of a retention time of about 221 minutes.

[0144] Separation method 1

[0145] Equipment used: YMC LC-Forte R

[0146] Chromatographic column: Waters Delta-Pak C-18RP HPLC column (30.0×300mm, 15μm)

[0147] Solvent: (a) Acetonitrile containing 0.02% TFA

[0148] (b) Water containing 0.02% TFA

[0149] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 40:60

[0150] From 0 to 60 minutes, the proportion of (a) solvent was increased from 40% to 100%.

[0151] Mobile phase flow rate: 10.0 mL / min

[0152] Detector: UV 254nm; ELSD

[0153] [Example 5] Isolation of Compounds 2 to 5 from Cassia Seed Bud Extract

[0154] (1) Isolation of compound 3

[0155] After the STS-EA-fraction 12 produced in the above-mentioned Example 3 (the second of the three fractions in which the elution solvent in Example 3 was dichloromethane (or ethyl acetate): methanol (or ethanol) in a mixing ratio of 10:1) was concentrated under reduced pressure, chromatography for separation was performed under the conditions of separation method 1 in the above-mentioned Example 4, and 1.2 mg of compound 3 having the structure of the above-mentioned chemical formula 3 was separated under the conditions of a retention time of about 24 minutes.

[0156] (2) Isolation of compound 2

[0157] After the STS-EA-fraction 8 of the above-mentioned Example 3 (the first of three fractions in which the elution solvent in Example 3 was dichloromethane (or ethyl acetate): methanol (or ethanol) in a mixing ratio of 50:1) was concentrated under reduced pressure, chromatography for separation was performed under the conditions of the following separation method 2, and 7.5 mg of compound 2 having the structure of the above-mentioned chemical formula 2 was separated under the conditions of a retention time of about 36 minutes.

[0158] Separation method 2

[0159] Equipment used: YMC LC-Forte R

[0160] Chromatographic column: Waters Delta-Pak C-18RP HPLC column (30.0×300mm, 15μm)

[0161] Solvent: (a) Acetonitrile containing 0.02% TFA

[0162] (b) Water containing 0.02% TFA

[0163] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 60:40

[0164] From 0 to 60 minutes, the proportion of (a) solvent was increased from 60% to 100%.

[0165] Mobile phase flow rate: 10.0 mL / min

[0166] Detector: UV 254nm; ELSD

[0167] (3) Isolation of compounds 4 and 5

[0168] After the STS-EA-fraction 13 of the above-mentioned Example 3 (the elution solvent in Example 3 is the third of the three fractions in which the mixing ratio of dichloromethane (or ethyl acetate): methanol (or ethanol) is 10:1) was concentrated under reduced pressure, the chromatography for separation was carried out under the conditions of the following separation method 3, and 0.6 mg of compound 4 having the structure of the above-mentioned chemical formula 4 was separated at a retention time of about 71 minutes, and 2.6 mg of compound 5 having the structure of the above-mentioned chemical formula 5 was separated at a retention time of about 120 minutes.

[0169] Separation method 3

[0170] Equipment used: JASCO LC-2000PLUS

[0171] Chromatographic column: Phenomenex Luna C-18RP HPLC column (21.2×250mm, 10μm)

[0172] Solvent: (a) Acetonitrile containing 0.02% TFA

[0173] (b) Water containing 0.02% TFA

[0174] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 10:90

[0175] From 0 to 60 minutes, the proportion of (a) solvent was increased from 10% to 50%.

[0176] From 60 minutes to 80 minutes, the proportion of (a) solvent was increased from 50% to 51%.

[0177] From 80 minutes to 140 minutes, the proportion of (a) solvent was increased from 51% to 58%.

[0178] From 140 minutes to 180 minutes, the proportion of (a) solvent was increased from 58% to 100%.

[0179] Mobile phase flow rate: 8 mL / min

[0180] Detector: UV 254nm

[0181] [Comparative Example 2] Isolation of Compounds A to X from Cassia Seed Bud Extract

[0182] (1) Separation of compounds F, G and H

[0183] After the STS-EA-fraction 12 of Example 3 was concentrated under reduced pressure, the fraction was subjected to chromatography for separation under the conditions of the separation method 1 of Example 4 to separate the components. As a result, 0.8 mg of Figure 4 Peak F is compound F, 0.9 mg equivalent to Figure 4 Peak G of compound G, 0.5 mg is equivalent to Figure 4 Peak H is of compound H.

[0184] (2) Separation of compounds C, D and E

[0185] After the STS-EA-fraction 8 of Example 3 was concentrated under reduced pressure, the components contained in the fraction were separated by chromatography under the conditions of the separation method 2 of Example 4. As a result, 7.2 mg of the equivalent of Figure 4 Peak C is compound C, 0.8 mg equivalent to Figure 4 Peak D is compound D, 2.5 mg equivalent to Figure 4 Peak E is compound E.

[0186] (3) Isolation of Compound I

[0187] After the STS-EA-fraction 15 obtained in Example 3 (the second fraction among the three fractions in which the elution solvent in Example 3 was dichloromethane (or ethyl acetate): methanol (or ethanol) in a mixing ratio of 5:1) was concentrated under reduced pressure, the components contained in the above fraction were separated by HPLC separation under the conditions of the following separation method 4. As a result, 0.7 mg of the equivalent of Figure 4 Compound I in Peak I of .

[0188] Separation method 4

[0189] Equipment used: JASCO LC-2000PLUS

[0190] Chromatographic column: Phenomenex Luna C-18RP HPLC column (21.2×250mm, 10μm)

[0191] Solvent: (a) Acetonitrile containing 0.02% TFA

[0192] (b) Water containing 0.02% TFA

[0193] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 30:70

[0194] From 0 to 240 minutes, the ratio of (a) solvent was maintained at 30%.

[0195] Mobile phase flow rate: 8 mL / min

[0196] Detector: UV 254nm

[0197] (4) Separation of compounds J, K, L, M, N, O, A and P

[0198] The STS-EA-fraction 16 obtained in the above-mentioned Example 3 (the third fraction among the three fractions in which the elution solvent is dichloromethane (or ethyl acetate): methanol (or ethanol) in a mixing ratio of 5:1) is concentrated under reduced pressure and then subjected to silica gel column chromatography under the conditions of the following separation method 5, thereby being graded into 14 sub-fractions.

[0199] Separation method 5

[0200] Equipment used: Rapid silica gel column

[0201] Chromatographic column: glass column (25.0×200mm)

[0202] Solvent: (a) dichloromethane (methylene chloride)

[0203] (b) Methanol

[0204] Mobile phase composition: start elution with a volume ratio of (a) solvent: (b) solvent of 20:1

[0205] The volume ratio of (a) solvent: (b) solvent was changed as follows: 10:1, 7:1, 4:1, 2:1, 1:1. The amount of solvent was: 300 mL.

[0206] From the above subfraction 3 (the second of the three subfractions in which the elution solvent in the above separation method 5 was dichloromethane:methanol in a mixing ratio of 10:1), 1.8 mg of the equivalent of Figure 4 Peak J is compound J, 1.9 mg equivalent to Figure 4 Peak K is compound K, 1.0 mg is equivalent to Figure 4 Peak L is compound L.

[0207] Separation method 6

[0208] Equipment used: Gilson 321HPLC

[0209] Chromatographic column: Phenomenex Luna C-18RP HPLC column (21.2×250mm, 10μm)

[0210] Solvent: (a) Acetonitrile containing 0.02% TFA

[0211] (b) Water containing 0.02% TFA

[0212] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 20:80

[0213] From 0 to 80 minutes, the proportion of (a) solvent was increased from 20% to 40%.

[0214] From 80 minutes to 120 minutes, the proportion of (a) solvent was increased from 40% to 100%.

[0215] Mobile phase flow rate: 8 mL / min

[0216] Detector: UV 254nm

[0217] In addition, from the above subfraction 4 (the third of the three subfractions in which the elution solvent in the above separation method 5 was dichloromethane:methanol in a mixing ratio of 10:1), 2.5 mg of the equivalent of Figure 4 Peak M is compound M, 2.1 mg equivalent Figure 4 Peak N is compound N, 0.5 mg is equivalent to Figure 4 The peak O is compound O.

[0218] Separation method 7

[0219] Equipment used: Gilson 321HPLC

[0220] Chromatographic column: Phenomenex Luna C-18RP HPLC column (21.2×250mm, 10μm)

[0221] Solvent: (a) Acetonitrile containing 0.02% TFA

[0222] (b) Water containing 0.02% TFA

[0223] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 20:80

[0224] From 0 to 100 minutes, the proportion of (a) solvent was increased from 20% to 80%.

[0225] From 100 minutes to 160 minutes, the proportion of (a) solvent was increased from 80% to 100%.

[0226] Mobile phase flow rate: 8 mL / min

[0227] Detector: UV 254nm

[0228] From the above subfraction 6 (the second of three subfractions in which the elution solvent in the above separation method 5 was dichloromethane:methanol in a mixing ratio of 7:1), 2.0 mg of the equivalent of Figure 4 Peak A is compound A.

[0229] In addition, from the above subfraction 7 (the third of the three subfractions in which the elution solvent in the above separation method 5 was dichloromethane:methanol in a mixing ratio of 7:1), 4.1 mg of the equivalent of Figure 4 Peak P is compound P.

[0230] Separation method 8

[0231] Equipment used: Gilson 321HPLC

[0232] Chromatographic column: Phenomenex Luna C-18RP HPLC column (21.2×250mm, 10μm)

[0233] Solvent: (a) Acetonitrile containing 0.02% TFA

[0234] (b) Water containing 0.02% TFA

[0235] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 17:83

[0236] From 0 to 50 minutes, the ratio of (a) solvent was maintained at 17%.

[0237] From 50 minutes to 70 minutes, the proportion of (a) solvent was increased from 17% to 25%.

[0238] From 70 minutes to 150 minutes, the proportion of (a) solvent was increased from 25% to 30%

[0239] From 150 minutes to 180 minutes, the proportion of (a) solvent was increased from 30% to 100%.

[0240] Mobile phase flow rate: 6.5 mL / min

[0241] Detector: UV 254nm

[0242] (5) Isolation of compound Q

[0243] After the STS-fraction 17 obtained in Example 3 (the first of three fractions in which the elution solvent in Example 3 was a mixture of dichloromethane (or ethyl acetate): methanol (or ethanol) in a ratio of 3:1) was concentrated under reduced pressure, the components contained in the above fraction were separated by HPLC separation under the conditions of the following separation method 9. As a result, 1.7 mg of the equivalent of Figure 4 Peak Q is compound Q.

[0244] Separation method 9

[0245] Equipment used: JASCO LC-2000PLUS

[0246] Chromatographic column: Phenomenex Luna C-18RP HPLC column (21.2×250mm, 10μm)

[0247] Solvent: (a) Acetonitrile containing 0.02% TFA

[0248] (b) Water containing 0.02% TFA

[0249] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 10:90

[0250] From 0 to 40 minutes, the ratio of (a) solvent was maintained at 10%.

[0251] From 40 minutes to 70 minutes, the proportion of (a) solvent was increased from 10% to 18%.

[0252] From 70 minutes to 180 minutes, the proportion of (a) solvent was increased from 18% to 22%.

[0253] From 180 minutes to 260 minutes, the proportion of (a) solvent was increased from 22% to 100%.

[0254] Mobile phase flow rate: 6.5 mL / min

[0255] Detector: UV 254nm

[0256] (6) Separation of compounds B, R, S, T, U, V, W and X

[0257] The STS-EA-fraction 18 obtained in the above-mentioned Example 3 (the second fraction of three fractions in which the elution solvent is dichloromethane (or ethyl acetate): methanol (or ethanol) in a mixing ratio of 3:1) was concentrated under reduced pressure and then subjected to silica gel column chromatography under the conditions of the following separation method 10, thereby being graded into 24 sub-fractions.

[0258] Separation method 10

[0259] Equipment used: Rapid silica gel column

[0260] Chromatographic column: glass column (25.0×200mm)

[0261] Solvent: (a) dichloromethane (methylene chloride)

[0262] (b) Methanol

[0263] Mobile phase composition: (a) solvent: (b) solvent volume ratio of 20:1 to start elution

[0264] The volume ratio of (a) solvent: (b) solvent was varied as 10:1, 7:1, 5:1, 3:1, 2:1, and 1:1

[0265] Step solvent volume: 400mL

[0266] 10:1 (subfractions 3, 4, 5, 6), 7:1 (subfractions 7, 8, 9, 10), 5:1 (subfractions 11, 12, 13, 14), 3:1 (subfractions 15, 16, 17, 18), 2:1 (subfractions 19, 20, 21, 22)

[0267] The above subfractions 17 to 21 (the third and fourth fractions of the four subfractions in the above separation method 10 in which the elution solvent was dichloromethane:methanol in a mixing ratio of 3:1, and the second to fourth fractions of the four subfractions in which the elution solvent was dichloromethane:methanol in a mixing ratio of 2:1) were separated by HPLC under the conditions of the following separation method 11. As a result, 1.6 mg of the equivalent of Figure 4 Peak B is compound B, equivalent to 24.8 mg Figure 4 Peak R of compound R, 5.5 mg is equivalent to Figure 4 Peak S is compound S, 2.3 mg equivalent to Figure 4 Peak T is compound T, 9.3 mg equivalent to Figure 4 Peak U is compound U, 6.4 mg equivalent Figure 4 Peak V of compound V, 4.1 mg equivalent Figure 4 Peak W is compound W, 0.6 mg is equivalent to Figure 4 Peak X is compound X.

[0268] Separation method 11

[0269] Equipment used: JASCO LC-2000PLUS

[0270] Chromatographic column: Phenomenex Luna C-18RP HPLC column (21.2×250mm, 10μm)

[0271] Solvent: (a) Acetonitrile containing 0.02% TFA

[0272] (b) Water containing 0.02% TFA

[0273] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 10:90

[0274] From 0 to 40 minutes, the ratio of (a) solvent was maintained at 10%.

[0275] From 40 minutes to 70 minutes, the proportion of (a) solvent was increased from 10% to 18%.

[0276] From 70 minutes to 180 minutes, the proportion of (a) solvent was increased from 18% to 22%.

[0277] From 180 minutes to 260 minutes, the proportion of (a) solvent was increased from 22% to 100%.

[0278] Mobile phase flow rate: 6.5 mL / min

[0279] The HPLC peaks corresponding to all the substances (Compounds 1 to 5, and Compounds C to X) isolated from Examples 4 and 5 and Comparative Example 2 are as follows: Figure 4 Each peak of the HPLC chromatogram of the Cassia seed bud extract is shown in the mark. A total of 29 substances including naphthopyrone derivatives (Compounds 1 to 5) having the structures of the above Chemical Formulas 1 to 5 are components isolated from the Cassia seed bud extract in the amounts obtained in the above Example 1. In addition, according to the HPLC analysis chromatogram, the above 29 substances are equivalent to the main constituent components of the Cassia seed bud extract.

[0280] [Example 6] Structural Identification of Compounds 1 to 5 Isolated from Cassia Seed Bud Extract

[0281] In order to identify the structures of Compounds 1 to 5 isolated in Examples 4 and 5, each compound was analyzed using a mass spectrometer (MS) and a nuclear magnetic resonance (NMR).

[0282] (1) Identification of compound 1

[0283] The molecular weight of the compound 1 isolated in Example 4 was determined to be 664 by MS measurement using an Agilent 1100 high performance liquid chromatography-mass spectrometer (HPLC-ESI-MS). The molecular weight of the compound 1 isolated in Example 4 was determined to be 664 by ultraviolet (PerkinElmer 343 Polarimeter), infrared (Thermo Scientific Nicolet iS50), specific rotation (PerkinElmer Lambda 35) spectral data, and nuclear magnetic resonance (Bruker 400MHz, 100MHz NMR) spectrometry. 1 H and 13 C NMR spectrum analysis confirmed that the compound 1 was 7-hydroxy-somyosomycin-erythromycin-8'-O-pyranoglucoside, which is a naphthopyrone derivative having the structure of the following chemical formula 1. The compound 1 is a novel compound that has not been reported so far and is isolated only as one of the main components in Cassia seed buds. This is because it is not contained in Cassia seed or fully grown Cassia seed plants or is contained in very small amounts and has not been found.

[0284] [Chemical formula 1]

[0285]

[0286] Molecular formula C 34 H 32 O 14 ; ESI-MS: m / z 665[M+H] + ; Infrared absorption band v max 3384, 2936, 1631, 1373, 1204, 1059cm -1 ; UV absorption band (MeOH)λ max (logε)240(4.3),282(4.2);Specific rotation[α] 22 D -20 (c 0.05, MeOH);

[0287] 1 H NMR (methanol-d4, 400MHz): δ7.18 (1H, s, H-10), 6.97 (1H, d, J = 2.0Hz, H-5), 6.92 (1H, s, H-9), 6.34 (1H, br s, H-5'), 6.12 (1H, s, H-3), 5.12 (1H, dd, J=7.5, 3.5Hz, H-1"), 3.98 (1H, dd, J=12.0, 2. 0Hz, H-6”a), 3.78 (1H, dd, J=12.0, 5.5Hz, H-6”b), 3.75 (3H, s, OMe-8), 3.58 (1H, t, J=8. 0Hz, H-2'), 3.55 (1H, dd, J=5.0, 2.0Hz, H-5"), 3.51 (1H, overlapped, H-3"), 3.48 (1H, t, J=9.0Hz, H-4”), 2.63 (3H, s, Me-10’), 2.41 (3H, s, Me-11), 1.98 (3H, s, Me-11’), 13C NMR (CDCl3, 500MHz): δ208.1(C-9'), 184.1(C-4), 170.1(C-2), 161.3(C-8), 160.9(C-5), 156.2(C-6'), 156.1(C-8'), 154. 7(C-6), 152.4(C-10a), 150.9(C-1'), 139.8(C-9a), 137.1(C-4'a), 132.4(C-3'), 122.4(C-2'), 120.6(C-4'), 111.9(C-7), 108.4(C-8'a), 106.2(C-5a), 105.8(C-3), 103.1(C-1”), 102.9(C-5’), 102.8(C-7’), 102.5(C-4a), 101.4(C-10), 97.0(C- 9), 77.4(C-5”), 76.7(C-3”), 73.5(C-2”), 69.7(C-4”), 61.0(C-6”), 54.8(OMe-8), 31.4(C-10’), 19.4(C-11), 16.1(C-11’)

[0288] (2) Identification of compound 2

[0289] The molecular weight of the compound 2 isolated in Example 5 was determined to be 272 by MS measurement using an Agilent 1100 high performance liquid chromatography-mass spectrometer (HPLC-ESI-MS). 1 H NMR spectrum analysis indicated that the product was isotoralactone, a naphthopyrone derivative having a structure shown in the following chemical formula 2. In addition, the NMR data was compared with the data in the existing literature (Kitanaka, S. et al. Phytochemistry, 1981, 20, 1951-1953) to identify its structure.

[0290] [Chemical formula 2]

[0291]

[0292] Molecular formula C 15 H 12 O5; ESI-MS: m / z 273[M+H] + ;

[0293] 1H NMR (CDCl3, 400MHz): δ13.30 (1H, s, OH-10), 9.41 (1H, s, OH-9), 6.92 (1H, s, H-5), 6.59 (1H, d, J = 2.5Hz, H-6), 6.56 ( 1H, d, J = 2.0Hz, H-8), 4.93 (1H, d, J = 2.0Hz, H-11), 4.62 (1H, d, J = 2.0Hz, H-11), 3.90 (3H, s, OMe-7), 3.80 (1H, s, H-4)

[0294] (3) Identification of compound 3

[0295] The molecular weight of the compound 3 separated in Example 5 was determined to be 272 by MS measurement using an Agilent 1100 high performance liquid chromatography-mass spectrometer (HPLC-ESI-MS). 1 H NMR spectrum analysis inferred that its structure was toralactone, a naphthopyrone derivative, having a structure as shown in the following chemical formula 3. In addition, the NMR data was compared with the data in the existing literature (Newman, AG et al. J Am Chem Soc 2016, 138, 4219-4228) to identify its structure.

[0296] [Chemical formula 3]

[0297]

[0298] Molecular formula C 15 H 12 O5; ESI-MS: m / z 273[M+H] + ;

[0299] 1 H NMR (CDCl3, 400MHz): δ13.56 (1H, s, OH-5), 9.44 (1H, s, OH-9), 7.00 (1H, s, H-5), 6.64 (1H, d, J=2 .5Hz, H-6), 6.55 (1H, d, J = 2.5Hz, H-8), 6.25 (1H, s, H-4), 3.92 (3H, s, OMe-7), 2.28 (3H, s, Me-11)

[0300] (4) Identification of compound 4

[0301] The molecular weight of the compound 4 separated in Example 5 was determined to be 288 by MS measurement using an Agilent 1100 high performance liquid chromatography-mass spectrometer (HPLC-ESI-MS). 1 The structure of the product was inferred to be torosachrysone, a naphthopyrone derivative, as shown in the following chemical formula 4 by H NMR spectrum analysis. In addition, the structure was identified by comparing the NMR data with the data in the existing literature (Gill, M. et al. Aust J Chem 2000, 53, 213-220).

[0302] [Chemical formula 4]

[0303]

[0304] Molecular formula C 16 H 16 O5; ESI-MS: m / z 289[M+H] + ;

[0305] 1 H NMR (methanol-d4, 400MHz): δ9.80 (1H, s, OH-9), 6.90 (1H, s, H-5), 6.57 (1H, d, J = 2.0Hz, H-6), 6.51 (1H, d, J = 2.0Hz, H-8), 3.91 (3H, s, OMe-7), 3.08 (2H, br s, H-4), 2.86 (2H, br s, H-2), 1.44 (3H, s, Me-11)

[0306] (5) Identification of compound 5

[0307] The molecular weight of the compound 5 separated in Example 5 was determined to be 288 by MS measurement using an Agilent 1100 high performance liquid chromatography-mass spectrometer (HPLC-ESI-MS). 1 The structure of the compound was inferred to be rubrofusarin, a naphthopyrone derivative, as shown in the following chemical formula 5 by H NMR spectrum analysis. The structure was also identified by comparing the NMR data with the data in the existing literature (Alemayehu, G. et al. Phytochemistry, 1993, 32, 1273-1277).

[0308] [Chemical formula 5]

[0309]

[0310] Molecular formula C 15 H 12 O5; ESI-MS: m / z 273[M+H] + ;

[0311] 1 H NMR (CDCl3, 400MHz): δ9.68 (1H, s, OH-9), 7.00 (1H, s, H-10), 6.59 (1H, d, J = 2.5Hz, H-9 ), 6.48 (1H, d, J = 2.0Hz, H-7), 6.04 (1H, s, H-3), 3.91 (3H, s, OMe-7), 2.41 (3H, s, Me-11)

[0312] [Test Example 1] Evaluation of the Antioxidant Effect of Cassia Seed Bud Extract

[0313] (1) Comparison of DPPH free radical scavenging effects of Cassia seed extract and Cassia seed bud extract

[0314] The Cassia seed extract (ST) prepared in the above-mentioned Comparative Example 1 and the Cassia seed bud extract (STS) prepared in the above-mentioned Example 1 were dissolved in methanol at concentrations of 1000ppm, 500ppm, 250ppm, and 125ppm, respectively. Then, 100μL of the Cassia seed extract solution of the above-mentioned Comparative Example 1 and 100μL of the Cassia seed bud extract of the above-mentioned Example 1 were reacted with 100μL of DPPH in the dark for 40 minutes, and the absorbance was measured at 517nm. As a positive control group, ascorbic acid (L-ascorbic acid) was used. The DPPH free radical scavenging effect obtained by the above-mentioned absorbance measurement and the following formula is shown in Figure 1a and Figure 1b .

[0315]

[0316] like Figure 1a and Figure 1b As shown, it can be seen that the DPPH radical scavenging effect of the Cassia seed bud extract (STS) of Example 1 is more than 1.7 times higher than that of the Cassia seed extract (ST) of Comparative Example 1 at all concentrations.

[0317] (2) Comparison of ABTS on-line antioxidant chromatography results of Cassia seed extract and Cassia seed bud extract

[0318] The Cassia seed extract (ST) produced in the above-mentioned Comparative Example 1 and the Cassia seed bud extract (STS) produced in the above-mentioned Example 1 were dissolved in an aqueous alcohol solution at the same concentration, and ABTS on-line antioxidant high performance liquid chromatography (HPLC) was performed under the following analysis conditions.

[0319] ABTS online antioxidant HPLC analysis conditions

[0320] Equipment used: Agilent 1200 system

[0321] Chromatographic column: RP C-18 HPLC column (4.6×150mm, 5μm)

[0322] Solvent: (a) Acetonitrile containing 0.02% TFA (Trifluoroacetic acid)

[0323] (b) Water containing 0.02% TFA

[0324] Mobile phase composition: elution started at 0 minutes with a volume ratio of (a) solvent: (b) solvent of 10:90

[0325] From 0 to 30 minutes, the proportion of (a) solvent was increased from 10% to 100%.

[0326] From 30 minutes to 37 minutes, elution was performed with 100% of (a) solvent.

[0327] Mobile phase flow rate: 0.7 mL / min

[0328] ABTS composition: water containing 0.08 mM ABTS and 0.12 mM potassium persulfate

[0329] ABTS flow rate: 0.35 mL / min

[0330] Detection wavelength: UV 254nm; 734nm

[0331] As a result, Cassia seed extract (ST) and Cassia seed bud extract (STS) showed very different antioxidant component patterns. Components composed of flavonoids and naphthalene derivatives with antioxidant activity were observed in both Cassia seed extract (ST) and Cassia seed bud extract (STS) before the retention time of 15 minutes, but in Cassia seed bud extract (STS), the content of the above components increased compared with Cassia seed extract (ST), and naphthalene and anthraquinone derivatives with antioxidant activity were further observed after 15 minutes. Figure 2a and Figure 2b The results of ABTS online antioxidant HPLC on Cassia seed and Cassia seed bud extracts are chromatograms obtained by detecting wavelengths of 254 nm (the chromatogram with the blue line marked upward) and 734 nm (the chromatogram with the red line marked downward) using an ultraviolet (UV) detector.

[0332] [Test Example 2] Comparison of ABTS on-line antioxidant chromatography results of Cassia seed extract (ST) and Cassia seed bud extract cultivated under various light conditions

[0333] The Cassia seed bud extract (STS) produced in the above Example 1 and the seven Cassia seed bud extracts (STS-C, STS-385, STS-465, STS-645 and STS-780) cultivated under various light conditions produced in the above Example 2 were dissolved in an alcohol aqueous solution, and ABTS on-line antioxidant high performance liquid chromatography (HPLC) was performed under the same analysis conditions as in the above Test Example 1.

[0334] As a result, components having antioxidant activity were commonly observed in the five types of Cassia seed bud extracts cultivated under various light conditions in Example 2, but the content of the components varied depending on the light conditions. Figures 3a to 3f The results of ABTS online antioxidant HPLC were respectively performed on the STS, STS-C, STS-385, STS-465, STS-645, and STS-780 extracts, which are the above-mentioned 6 types of Cassia seed bud extracts. The chromatograms were obtained by detecting wavelengths of 254 nm (indicating the peak of components that absorb UV at 254 nm) and 734 nm (the degree of antioxidant effect is indicated by the peak of components that show antioxidant effects on ABTS and the area of ​​the peak) using an ultraviolet (UV) detector.

[0335] based on Figures 3a to 3f The chromatograms of the antioxidant components of Cassia seed bud extracts cultivated under light-shielding conditions, fluorescent lights, and light conditions of specific wavelengths were used. The content and composition of antioxidant components varied depending on the wavelength of light irradiated during the cultivation process, but no components were found to disappear or appear under specific conditions. Figure 3a to Figure 3f In all the chromatograms, the novel naphthopyrone component having the structure of the above chemical formula 1 of the present invention (compound 1 separated in Example 4) was detected as the most significant antioxidant peak or the main antioxidant peak, and the four naphthopyrone components having the structures of the above chemical formulas 2 to 5 (compounds 2 to 5 separated in Example 5) were also detected in Figure 3a to Figure 3f In addition to the chromatogram of Cassia seed bud extract (STS-C) cultivated under normal light conditions ( Figure 3b ) and the chromatogram of Cassia seed bud extract (STS-465) cultivated under 465nm LED illumination ( Figure 3d ) Figure 3a )、( Figure 3c )、( Figure 3e )and( Figure 3f ) chromatogram, relatively high contents of the active ingredients of compounds 1 to 5 separated in the above Examples 4 and 5 were detected.

[0336] [Test Example 3] Protective effects of seven Cassia seed bud extracts from Examples 1 and 2 on retinal progenitor cells (R28)

[0337] In order to confirm the protective effects of the Cassia seed extract (ST) and Cassia seed bud extract (STS) obtained in the above-mentioned Comparative Example 1 and Example 1, respectively, and the Cassia seed bud extracts (STS-C, STS-385, STS-465, STS-645, STS-780) cultivated under various light conditions obtained in the above-mentioned Example 2 on retinal progenitor cells (R28), the following experiment was performed.

[0338] Specifically, R28 cells were cultured in a DMEM (Dulbecco's modified eagle's medium) / low glucose medium containing 10% FBS (fetal bovine serum), 100 U / mL penicillin, and 100 μg / mL streptomycin in a 37°C incubator under 5% CO2 conditions, and were subcultured every 2 days using 0.05% trypsin, and then plated in a 96-well plate at 1×10 4 The seven extracts were inoculated at a density of 1.5 and cultured for 24 hours. Figure 5a to Figure 5c The R28 cells were treated with 10 μL of ez-cy tox at a concentration of 10 μL. After 2 hours, 10 mM glutamate and 0.5 mM BSO (buthionine sulphoximine) were added and cultured for 22 hours. Then, in order to measure the cell survival rate, 10 μL of EZ-cy tox was treated for 2 hours, and UV absorbance was measured at 450 nm.

[0339] Figure 5a The results are expressed by cell survival rate of the effects of the Cassia seed extract (ST) of Comparative Example 1, the Cassia seed bud extract (STS) of Example 1, and the five Cassia seed bud extracts (STS-C, STS-385, STS-465, STS-645, and STS-780) cultivated at three different concentrations under different light conditions of Example 2 to protect R28 cells from oxidative stress induced by glutamate. Figure 5b and Figure 5c The results are expressed in terms of cell survival rate, in terms of the protective effects of the Cassia seed extract (ST) of Comparative Example 1 and the Cassia seed bud extract (STS) of Example 1 on R28 cells against glutamate-induced oxidative stress at five different concentrations.

[0340] like Figure 5a As shown in the results, all the extract treatment groups showed the effect of protecting HT-22 cells from oxidative stress induced by glutamate, but compared with the Cassia seed extract, the Cassia seed bud extracts (STS, STS-C, STS-385, STS-465, STS-645 and STS-780) showed a better effect of protecting R28 cells from oxidative stress induced by glutamate in most concentrations, especially, as Figure 5b and Figure 5c As shown, it was confirmed that the Cassia seed bud extract (STS) of Example 1 was significantly superior to the Cassia seed extract (ST) of Comparative Example 1 in protecting R28 cells.

[0341] [Test Example 4] Confirmation of the protective effect of compounds 1 to 5 isolated from the Cassia seed bud extracts of Examples 5 and 6 on retinal progenitor cells (R28)

[0342] In order to confirm why the protective effect of the Cassia seed bud extract (STS) of Test Example 3 on retinal progenitor cells (R28) is superior to that of the Cassia seed extract (ST), the protective effects of Compounds 1 to 5 of Examples 5 and 6 on R28 cells were compared.

[0343] The survival rate of R28 cells was determined by the same method as in Experiment 3. In addition, instead of the Cassia seed extract (ST) and Cassia seed bud extract (STS, STS-C, STS-385, STS-465, STS-645 and STS-780) of Comparative Example 1 and Examples 1 and 2, compounds 1 to 5 of Examples 5 and 6 were treated with 50 μM, 16.6 μM and 5.55 μM, respectively. The results are shown in Figure 6a and Figure 6b .

[0344] like Figure 6a and Figure 6b As shown, compounds 1 to 5 are effective in protecting R28 cells from glutamate-induced neurotoxicity. The above compounds are relatively high in content or mainly present in Cassia seed bud extract (STS), so it can be confirmed that the protective effect of Cassia seed bud extract (STS) on R28 cells is superior to that of Cassia seed extract (ST).

[0345] [Test Example 5] Protective Effects of Seven Cassia Seed Bud Extracts from Examples 1 and 2 on Hippocampal Neuronal Cells (HT-22)

[0346] In order to confirm the protective effect of Cassia seed extract (ST) and Cassia seed bud extract (STS) obtained in the above-mentioned Comparative Example 1 and Example 1, and Cassia seed bud extracts cultivated under various light conditions (STS-C, STS-385, STS-465, STS-645, STS-780) obtained in the above-mentioned Example 2 on hippocampal neurons (HT-22), the following experiment was performed.

[0347] Specifically, HT-22 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) / low-glucose medium containing 10% FBS (fetal bovine serum), 100 U / mL penicillin, and 100 μg / mL streptomycin in a 37.5°C incubator under 5% CO2 conditions, and were subcultured every 2 days using 0.05% trypsin. 3 The seven extracts were inoculated at a density of 1.5 and cultured for 24 hours. Figures 7a to 7c The HT-22 cells were treated with 10 μL of EZ-cytox for 2 hours and the UV absorbance was measured at 450 nm to measure the cell survival rate.

[0348] Figure 7a The results are as follows: the effects of five types of Cassia seed bud extracts (STS-C, STS-385, STS-465, STS-645, and STS-780) cultivated at three different concentrations under different light conditions of the Cassia seed extract (ST) of the above-mentioned Comparative Example 1, the Cassia seed bud extract (STS) of the above-mentioned Example 1, and the above-mentioned Example 2 on protecting HT-22 cells from oxidative stress induced by glutamate are expressed by cell survival rate. Figure 7b and Figure 7c The results are expressed in terms of cell survival rate, showing the protective effects of the Cassia seed extract (ST) of Comparative Example 1 and the Cassia seed bud extract (STS) of Example 1 at five different concentrations on HT-22 cells from oxidative stress induced by glutamate.

[0349] like Figure 7a As shown, in all the extract treatment groups, the effect of protecting HT-22 cells from oxidative stress induced by glutamate was shown, especially, Figure 7b and Figure 7c As shown, it was confirmed that the Cassia seed bud extract (STS) of Example 1 showed a superior HT-22 cell protective effect compared with the Cassia seed extract (ST) of Comparative Example 1.

[0350] [Test Example 6] Confirmation of the protective effect of compounds 1 to 5 isolated from the Cassia seed bud extracts of Examples 5 and 6 on hippocampal neurons (HT-22)

[0351] In order to confirm why the protective effect of the Cassia seed bud extract (STS) of the above-mentioned Test Example 5 on hippocampal neurons (HT-22) is superior to that of the Cassia seed extract (ST), the protective effects of compounds 1 to 5 of Examples 5 and 6 and compounds A to X of Comparative Example 2 on R28 cells were compared.

[0352] The survival rate of HT-22 cells was determined by the same method as in the above-mentioned Test Example 5, and instead of the Cassia seed extract (ST) and Cassia seed bud extract (STS, STS-C, STS-385, STS-465, STS-645 and STS-780) of Comparative Example 1, Examples 1 and 2, compounds 1 to 5 of Examples 5 and 6 were treated with 50 μM, 16.6 μM and 5.55 μM, and the results are shown in Figure 8a and Figure 8b .

[0353] like Figure 8a and Figure 8b As shown, the effects of Compound K and Compound T, which are respectively equivalent to Peak K and Peak T of Compounds 1 to 5 of Examples 5 and 6 and Comparative Example 2, on protecting HT-22 cells from glutamate-induced neurotoxicity were shown. That is, the above-mentioned Compounds 1 to 5 are compounds whose contents are relatively very high in the Cassiae bud extract (STS) or are mainly present in the Cassiae bud extract (STS), and therefore, it can be confirmed that the HT-22 cell protective effect of the Cassiae bud extract (STS) is superior to that of the Cassiae bud extract (ST).

[0354] The physiological activities of the components evaluated in the above-mentioned Test Examples 1, 4 and 6 are shown in the following Table 1. The following Table 1 relates to the physiological activities of the components separated from the Cassia seed sprout extract (STS) of Example 1.

[0355] Physiological activities of components isolated from Cassia seed bud extract

[0356]

[0357] O significant activity; X no activity; w weak activity.

Claims

1. A naphthopyrone derivative of the following chemical formula 1 or a pharmaceutically acceptable salt thereof, Chemical formula 1 2. A method for producing a naphthopyrone derivative or a pharmaceutically acceptable salt thereof, which is a method for producing a naphthopyrone derivative or a pharmaceutically acceptable salt thereof of the following chemical formula 1, comprising drying and crushing buds obtained by cultivating Cassia obtusifolia L. seeds under light-shielding conditions for 6 days, adding ethanol to obtain a bud extract of Cassia obtusifolia L. seeds, adding water and ethyl acetate to the bud extract of Cassia obtusifolia L. seeds, stirring and fractionating at room temperature to obtain an ethyl acetate fraction, contacting the ethyl acetate fraction with an elution solvent having a mixed volume ratio of dichloromethane:methanol of 10:1, stirring in diatomaceous earth, evaporating the solvent, developing with silicone resin, and obtaining three fractions by chromatography, and concentrating the second fraction of the three fractions under reduced pressure, and then separating the naphthopyrone derivative by chromatography, Chemical formula 1 3. A use of a naphthopyrone derivative, a pharmaceutically acceptable salt thereof, a bud extract of Cassia obtusifolia L. seeds containing the same, or a bud fraction of Cassia obtusifolia L. seeds containing the same for the manufacture of a composition for protecting nerve cells from oxidative stress or for inhibiting nerve cell death, wherein the naphthopyrone derivative is a naphthopyrone derivative of the following chemical formula 1, Chemical formula 1 4. The use according to claim 3, wherein The oxidative stress is induced by glutamate.

5. The use according to claim 3, wherein The nerve cells are retinal nerve cells or hippocampal nerve cells.

6. Use of a naphthopyrone derivative, a pharmaceutically acceptable salt thereof, a bud extract of Cassia obtusifolia L. seeds containing the same, or a bud fraction of Cassia obtusifolia L. seeds containing the same, for producing a composition for preventing or treating a neurological injury disease induced by damage or death of retinal neurons or hippocampal neurons, wherein the naphthopyrone derivative is a naphthopyrone derivative of the following chemical formula 1: Chemical formula 1 7. The use according to claim 6, wherein The neurological damage disease is one or more selected from memory loss, learning impairment, depression, amyotrophic lateral sclerosis, Parkinson's disease, cerebral ischemic nerve damage, diabetic neuropathy, macular degeneration, visual impairment caused by retinal cell damage, retinitis pigmentosa, retinal detachment, retinal vascular occlusion and glaucoma.

8. The use according to claim 6, wherein The damage or death of the retinal nerve cells or hippocampal nerve cells is caused by glutamate neurotoxicity.

9. The use according to claim 6, wherein The composition is a pharmaceutical composition.

10. Use of a naphthopyrone derivative, a pharmaceutically acceptable salt thereof, a bud extract of Cassia obtusifolia L. seeds containing the same, or a bud fraction of Cassia obtusifolia L. seeds containing the same for the manufacture of an antioxidant composition, wherein the naphthopyrone derivative is a naphthopyrone derivative of the following chemical formula 1: Chemical formula 1

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