JP2019064957A - Neuron elongation promoter - Google Patents

Abstract

To provide a neuron elongation promoter that can promote elongation of neurons.SOLUTION: A neuron elongation promoter has 6-methylsulfinyl hexyl isothiocyanate or a glycoside thereof, and at least one selected from the group of unsaturated fatty acid and polyphenol.SELECTED DRAWING: None

Description

  The present invention relates to a nerve cell elongation promoter.

  In recent years, in preparation for the aging society, research on dementia is in progress. There are various known causes of dementia, among which many are due to neuronal damage and degeneration. Neurons extend long neurites and are specialized cells to transmit stimulation information. Therefore, conventionally, medicines have been developed to suppress nerve cell damage and degeneration.

Unexamined-Japanese-Patent No. 2010-202559 JP, 2015-051927, A

The applicant has for many years developed a medicine and functional food using wasabi component (Patent Documents 1 and 2). Applicants have attempted to promote neuronal outgrowth using a horseradish component.
The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a nerve cell elongation promoting agent capable of promoting nerve cell elongation.

The nerve cell elongation promoter of the present invention comprises 6-methylsulfinylhexyl isothiocyanates or a glycoside thereof;
At least one member selected from the group consisting of unsaturated fatty acids and polyphenols;
Have.

  According to the nerve cell extension promoting agent of the present invention, the extension of nerve cells can be promoted.

It is explanatory drawing of a nerve cell. It is a figure which shows the extension test result of Experimental example 1. FIG. It is a figure which shows the toxicity test result of 6-methylsulfinyl hexyl isothiocyanate (6-MSITC) of Experimental example 2. It is a figure which shows the toxicity test result of the curcumin of Experimental example 2. FIG. It is a figure which shows the toxicity test result of DHA of Experimental example 2. It is a figure which shows the toxicity test result of the isorhamnetin of Experimental example 2.

  The nerve cell elongation promoter of this embodiment comprises (A) 6-methylsulfinylhexyl isothiocyanate (hereinafter also referred to as “6-MSITC”) or an isothiocyanate such as a glycoside thereof; And at least one selected from the group consisting of saturated fatty acids and (C) polyphenols.

(A) Isothiocyanates In this embodiment, isothiocyanates refer to a compound having a -NCS group. The isothiocyanates may have, for example, aliphatic or aromatic groups as side chains. As isothiocyanates having an aliphatic group, for example, isopropyl isothiocyanate, isobutyl isothiocyanate, 2-butyl isothiocyanate, isoamyl isothiocyanate, amyl isothiocyanate, allyl isothiocyanate, 3-butenyl isothiocyanate, 4-pentenyl Isothiocyanate, 5-hexenyl isothiocyanate, 6-heptenyl isothiocyanate, 3-methylthiopropyl isothiocyanate, 4-methylthiobutyl isothiocyanate, 5-methylthiopentyl isothiocyanate, 6-methylthiohexyl isothiocyanate, 7-methylthioheptyl isothiocyanate , 4-methylsulfinylbutyl isothiocyanate, 5-methylsulfinylpentyl isothiocyanate And 6-methylsulfinylhexyl isothiocyanate, 6-methylsulfinylhexyl isothiocyanates (6-MSITCs), and 7-methylsulfinylheptyl isothiocyanate, and one or more selected from the group consisting of Methyl sulfinyl butyl isothiocyanate, 5-methyl sulfinyl pentyl isothiocyanate, 6-MSITCs, 7-methyl sulfinyl heptyl isothiocyanate is preferable. Examples of isothiocyanates having an aromatic group include phenethyl isothiocyanate.

In the present embodiment, isothiocyanates are a concept also including glycosides.
In this embodiment, preferred among isothiocyanates are 6-MSITCs or their glycosides.

  In this embodiment, the 6-MSITCs are meant to include 6-MSITC and 6-MSITC analogs. 6-MSITC is represented by the following chemical formula.

  In this embodiment, 6-MSITC analogs include naturally occurring analogs of 6-methylsulfinylhexyl isothiocyanate, and analogs of non-naturally occurring 6-methylsulfinylhexyl isothiocyanate obtained by synthesis. For example, 6-methylthiohexyl isothiocyanate (sulfur is not oxidized, ie a structure having a methyl sulfide group) and 6-methylsulfonylhexyl isothiocyanate (sulfur is peroxidized, ie a structure containing a methylsulfonyl group), etc. Point to

  The 6-MSITCs used in the present embodiment are, for example, contained in plants of the family Brassicaceae. Among Brassicaceae, the present wasabi and / or horseradish contains many 6-MSITCs.

  The 6-MSITCs used in the present embodiment can be obtained by a chemical synthesis method, but may be extracted and purified from plants. Examples of plants that can be used as raw materials for 6-MSITCs include, for example, Bataceae (Bataceae), Brassicaceae (Brassicaceae), Bresch Neidaceae (Bretschneideraceae), Amaryngea (Capparaaceae), Papaya (Caricaceae), and Euphorbiaceae ( Euphorbiaceae), Gyrostemonaceae (Gyrostemonaceae), Limnantes (Limnanthaceae), Horseradish (Moringaceae), Pentadiplandracea (Pentadiplandraceae), Phytolactocae (Phytolaccaceae), Tobella (Pittosporaceae), Molybdaceae (Salv doraceae), Towaria family (Tovariaceae), Nouzenharen family (Tropaeolaceae), mention may be made of the plant or the like. Specifically, for example, wasabi (Wasabi japonica) [alias: this horseradish], horseradish (Armoracia rusticana) [alias: mountain wasabi], Batis maritima (Japanese name unknown), mustard (Brassica juncea), broccoli (Brassica) oleracea var. italica, Arabidopsis thaliana, Capsella bursa-pastoris, watercress (Nasturtium officinale), Bretschneidera sinensis (Japanese name unknown), caper (Capparis spinosa), papaya (Carica papaya rica) Unknown name); Putranjiva roxburghii (Japanese name unknown), Tersonia brevipes (Japanese name unknown), Limnanthes douglasii (Japanese name unknown), Wasabi tree (Moringa oleifera), Pentadiplandra brazzeana (Japanese name unknown), Phytolacca (Phytolacca americana) Bursaris spinosa var. Examples thereof include incana (Japanese name unknown), Shinobumokuso (Reseda alba), Salvadora persica (Japanese name unknown), Tovaria pendula (Japanese name unknown), Kinrenka (Tropaeolum majus) and the like. However, 6-MSITCs that can be used in the present disclosure are not limited to those obtained from the above-mentioned plants, and all natural resources containing 6-MSITCs can be used as a raw material.

  Examples of the method for extracting and purifying the plant include a method for extracting 6-MSITCs from wasabi and horseradish which are cruciferous plants. A preferred extraction method is disclosed by patent 3919489.

  In the present embodiment, the glycoside of 6-MSITCs is a compound in which a sugar residue is bound to 6-MSITCs. The sugar residue binding to 6-MSITCs may be one or more sugars selected from the group consisting of glucose, rhamnose, fructose and galactose, with glucose being more preferred. The sugar residue may be either monosaccharide or polysaccharide, but monosaccharide is preferred. A monosaccharide refers to a sugar consisting of monosaccharide molecules. A polysaccharide refers to a sugar chain to which two or more monosaccharide molecules are linked. The polysaccharide is preferably formed by combining 2 or more and 10 or less monosaccharides, and more preferably formed by combining 2 or more and 5 or less monosaccharides.

The bonding position of the sugar residue bonded to 6-MSITCs is preferably an NCS group moiety, and is further preferably bonded to S.
The glycoside of 6-MSITCs is, for example, glucohesperin. Glucohesperin is a compound represented by the following chemical formula.

  The glycosides of 6-MSITCs can be obtained from plants containing glycosides of 6-MSITCs, such as the present wasabi. For example, after the enzyme reaction is stopped by heating (including electromagnetic waves, far infrared rays, baking, etc.), drying, freezing, enzyme treatment, chemical treatment, etc., the plant is subjected to solvent extraction with water, an organic solvent, etc. The glycosides of 6-MSITCs are extracted or separated and purified by performing means such as pressing, drying, crushing, purification, enzyme reaction, chemical reaction and the like. In addition, glycosides of 6-MSITCs can also be obtained by chemical synthesis.

(B) Unsaturated fatty acids Unsaturated fatty acids refer to fatty acids having one or more double bonds. Unsaturated fatty acids having two or more double bonds are preferred. The carbon number of the unsaturated fatty acid is not particularly limited, but is preferably 12 to 24, more preferably 15 to 24, and preferably 18 to 22.

The position of the double bond possessed by the unsaturated fatty acid may be any one or more of the ω3 position, the ω6 position, and the ω9 position. In particular, the unsaturated fatty acid is preferably an ω3 fatty acid.
Omega 3 fatty acids refer to unsaturated fatty acids having a double bond in at least the ω 3 position of fatty acids. The ω3 fatty acids, for example, docosahexaenoic acid (DHA, 22: 6 (△ 4,7,10,13,16,19)), docosapentaenoic acid (DPA, 22: 5 (△ 7,10,13,16 ^{, 19} )), eicosapentaenoic acid (EPA, 20: 5 (Δ ^{5, 8, 11, 14 , 17} )), α-linolenic acid (ALA, 18: 3 (Δ ^{9, 12, 15} )). .

  Omega 3 fatty acids may be extracted from fish and / or fish oil. DHA is, for example, contained in blue fish. EPA is contained, for example, in fish oils such as cod, herring, mackerel, salmon, sardines and krill. Among them, DHA and EPA are preferred, and DHA is more desirable.

(C) Polyphenol In this embodiment, a polyphenol is a general term for a compound having a plurality of phenolic hydroxy groups in a molecule. The polyphenol may consist of one or more selected from the group of curcuminoids and flavonoids.

  The curcuminoid may be represented by the following formula (I).

(Wherein, Ar represents an aromatic group having an OH group)
Curcuminoids include curcumin dimethoxycurcumin and bisdimethoxycurcumin, of which curcumin is preferred. Among curcumins, those shown in the following formulas (II) to (I-IV) are exemplified.

Examples of the component having curcumin include turmeric extract, turmeric pigment, and turmeric pigment.
Curcuminoids may be extracted from turmeric. The curcuminoid containing curcumin can be obtained, for example, by an extraction separation method using an organic solvent, an alcohol solvent or the like with turmeric or the like as a raw material, or a synthesis method.

  The flavonoid is preferably a flavonol represented by the following formula (II).

(Wherein, X represents -O ^- or -OH, Y represents -O ^- or -OH, Z represents -O ^- or -OH, and X, Y and Z represent two or more in total. N represents an integer of 1 to 4, and m represents an integer of 1 to 5.)
As flavonols, those shown in the following formulas (II-I) to (II-XI) can be mentioned. Among these, isorhamnetin represented by the formula (II-III) is preferable.

As a component containing isorhamnetin, for example, ginkgo biloba (ginkgo) extract can be mentioned.
Flavonoids may be extracted from Ginkgo biloba. The flavonoid can be obtained by a known method from a plant containing flavonoid or a known synthetic method. Isorhamnetin can be extracted from Ginkgo leaves by known methods.

The nerve cell elongation promoting agent of the present embodiment may have 6-MSITCs, docosahexaenoic acid (DHA), curcumin and isorhamnetin alone or in combination of two or more.
The nerve cell elongation promoting agent of the present embodiment may have a combination of the following components.
6-MSITCs and DHA,
6-MSITCs and curcumin,
6-MSITCs and isorhamnetin.

  When the nerve cell extension promoter of the present embodiment has (A) 6-MSITCs and (B) docosahexaenoic acid (DHA), the molar ratio of (B) DHA to (A) 6-MSITCs is 0. The number is preferably 2 or more and 400 or less, preferably 0.2 or more and 80 or less, and more preferably 0.2 or more and 16 or less.

  When the nerve cell elongation promoting agent of the present embodiment has (A) 6-MSITCs and (CI) curcumin, the molar ratio of (CI) curcumin to (A) 6-MSITCs is 0.04 or more and 25 or less It is preferable that the number is 0.04 or more and 1 or less, and more preferably 0.2 or more and 1 or less.

  When the nerve cell extension promoter of the present embodiment has (A) 6-MSITCs and (CII) isorhamnetin, the molar ratio of (CII) isorhamnetin to (A) 6-MSITCs is 0.008 or more and 5 or less It is preferable that the ratio is 0.008 or more and 1.0 or less, and more preferably 0.008 or more and 0.2 or less.

In these cases, the effect of extending neurites can be promoted more effectively.
The dose of the nerve cell extension promoting agent of the present embodiment is determined by considering the age, sex, body weight, usage, dose and the like of the patient. As the usage, oral administration, vascular injection, external use, application and the like can be mentioned. In the case of oral administration, the daily dose of 6-MSITCs in the nerve cell elongation promoter is preferably 10 μg to 100 mg / day, and the dose of unsaturated fatty acid is preferably 100 mg / day to 10 g / day. The dosage may be 0.1 mg / day to 10 g / day.

  When the method of use of the nerve cell extension promoter of the present embodiment is vascular injection, the daily dose of 6-MSITCs in the nerve cell extension promoter is preferably 1 μg / day to 5 mg / day, and the unsaturated fatty acid The dose may be 1 mg / day to 100 mg / day, and the dose of polyphenol may be 0.1 μg / day to 1 mg / day.

  The nerve cell extension promoting agent of the present embodiment may further contain an additive. As the additive, for example, an excipient, a disintegrant, a binder, an antioxidant, a coating, a coloring agent, a flavoring agent, a surfactant, a plasticizer and the like can be blended.

In addition, anti-allergic agents, cooling agents, vitamins and other herbal medicines can be blended as long as the effects of the present invention are not impaired.
The nerve cell elongation promoting agent of the present embodiment may be contained in food, quasi-drugs and medicines.

  The form of the food containing the nerve cell extension promoter of the present embodiment is optional and is not limited. Specific food forms are, for example, general foods, general beverages, supplements, health foods, functional display foods, health functional foods such as foods for specified health and food for specific applications, soft drinks, tea drinks, drink agents And alcoholic beverages such as wine, confectionery, cooked rice, breads, noodles, vegetables, seasonings and the like.

Although the usage of the quasi-drugs and pharmaceuticals containing the nerve cell extension promoter of this embodiment is arbitrary and it is not limited, For example, an internal use * external preparation etc. are mentioned.
Although the dosage form of cosmetics and quasi-drugs containing the nerve cell extension promoter of this embodiment is not limited, for example, a capsule, a tablet, a powder, a granule, a liquid, etc. are mentioned.

  The nerve cell extension promoting agent of the present embodiment promotes the protrusion extension of nerve cells. The nerve cell extension promoting agent of the present embodiment can be used to prevent or treat a disease or condition involving a nerve cell.

  The nerve cell elongation promoter of this embodiment includes dementia, type 2 diabetes, Alzheimer's disease, diabetes, Parkinson's disease, transmissible spongy encephalopathy, so-called "mad cow disease", Huntington's disease, medullary thyroid carcinoma, arrhythmia, Arteriosclerosis, rheumatoid arthritis, aortic medial amyloid, prolactin producing tumor, familial amyloid polyneuropathy, hereditary non-neuropathic amyloidosis, dialysis amyloidosis, Finnish amyloidosis, lattice corneal dystrophy, cerebral amyloid angiopathy, systemic AL amyloidosis , Associated body myositis (Sporadic Inclusion Body Myositis), pheochromocytoma, osteomyelitis, multiple myeloma, encephalitis, meningitis, pre-Alzheimer's disease, mild cognitive impairment, early Alzheimer's disease, late Alzheimer's disease, related to aging Dementia, Parkinson's disease, Hunch Ton's disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease), prion disease, Creutzfeldt-Jakob disease, Lewy body disease, Friedreich's ataxia, stroke, genetic encephalopathy It is effective in improving symptoms such as schizophrenia, depression, bipolar disorder, attention deficit hyperactivity disorder (ADHD), autism, Asperger's syndrome, and Down's syndrome.

(Experimental example 1)
The protrusion extension test of PC12 cells using various components was performed as follows.
(1) Preparation of various components (a) 6-MSITC (component 1)
6-MSITC (manufactured by Gold Seal Co., Ltd.) was obtained. 6-MSITC used in this example was obtained by a chemical synthesis method. The chemical synthesis method of 6-MSITC is specifically described as follows.

In principle, the method of Kjaer et al. Is followed (Kjaer et al., Acta chem .. Scand., 11, 1298, 1957). Reflux with CH ₃ -SNa using 6-chlorohexanol to give 6-methylthiohexanol. This is reacted with thionyl chloride (SOCl ₂ ) to give 6-chlorohexyl methyl sulfide. Next, using the Gabriel method, an amino group is introduced with phthalimide potassium salt to form N- (6-methylthiohexyl) -phthalimide. Hydrazine hydrate is added thereto and refluxed to obtain 6-methylthiohexylamine, which is then reacted with thiocarbonyl chloride to obtain 6-methylthiohexyl isothiocyanate.

  Furthermore, the obtained 6-methylthiohexyl isothiocyanate is oxidized in methylthio group with m-chloroperbenzoic acid to obtain 6-methylsulfinylhexyl isothiocyanate (6-MSITC) (Morimitsu et. Al., J. Biol. Chem., 277, 3456, 2002).

In addition, 6-MSITC according to the present invention is contained in cruciferous plants and the like, and can also be obtained by solvent extraction or grinding of these plants.
100 mg of 6-MSITC was dissolved in DMSO (dimethyl sulfoxide) solvent to prepare a 6-MSITC solution with 100 mM 6-MSITC.
(B) DHA (component 2)
DHA (manufactured by Nacalai Tesque, product number 14122-64) was obtained. This DHA is extracted from blue fish. 100 mg DHA was dissolved in DMSO solvent to prepare DHA solution with 100 mM DHA.
(C) Curcumin (component 3)
Curcumin (CAS No. 458-37-7, manufactured by Tokyo Chemical Industry Co., Ltd., product No. C2302) was obtained. Curcumin is represented by the following chemical formula.

This curcumin is a synthetic one. 67.56 mg of curcumin was dissolved in DMSO solvent to prepare a curcumin solution with 200 mM curcumin.
(D) isorhamnetin (component 4)
Isolamnetin (manufactured by EXTRASYNTHESE, product number 1120S) was obtained. Isorhamnetin is extracted from ginkgo leaves. An isorhamnetin solution having 10 mM isorhamnetin was prepared by dissolving in DMSO solvent using a pipette for 10 mg isorhamnetin.

(2) Preparation of medium A growth medium A and a differentiation medium B containing the following components were prepared.
(A) Component of growth medium A DMEM (Dulbecco's Modified Eagle Medium, manufactured by Nacalai Tesque, product number 08458-45)
10 wt% horse serum (Thermo Fisher Scientific, product number 16050-122)
5 wt% bovine serum (Thermo Fisher Scientific, product number 10437-028)
1 wt% penicillin streptomycin (Nacalai Tesque, product number 26253-84)
(B) Component of differentiation medium B DMEM (Dulbecco's Modified Eagle Medium, manufactured by Nacalai Tesque, product number 08458-45)
2 wt% horse serum (Thermo Fisher Scientific, product number 16050-122)
1 wt% penicillin streptomycin (Nacalai Tesque, product number 26253-84)
(3) PC12 Cell Proliferation Cell growth flask A (manufactured by TPP) was charged with the above growth medium A, and further, PC12 cells (manufactured by RIKEN, product number RCB 0009) were added and proliferated. PC12 cells are pheochromocytomas from rat adrenal medulla and are models of neuronal differentiation. The PC12 cells were passaged by placing the grown PC12 cells in a new flask containing growth medium A in a new flask containing growth medium A when the confluence was 80-90%. The ratio of the culture solution in which the cells were grown to the new growth medium A was 1: 5 (volume ratio). Such passaging was repeated three times or more. Such passaging was repeated three times or more.

(4) Differentiation of PC12 cells 100 μl of growth medium A containing 5 × 10 ⁴ / ml of PC12 cells after passage obtained in (3) was seeded in each well of a 96-well plate. Leave at room temperature overnight to stabilize PC12 cells. The plate was spun to sediment PC12 cells in each well of the plate. The supernatant was removed. In each well in which PC12 cells were precipitated, 100 μl of differentiation medium B was added and stirred to suspend PC12 cells in differentiation medium B.

NGF (Sigma-Aldrich, product number N 2513) was added to the medium in each well. The concentration of NGF contained in the media in all the wells was 50 ng / ml.
Furthermore, the various components 1 to 4 prepared in (1) were added to the culture medium in each well so as to have various concentrations in one type or in a combination of two types, to obtain samples 1 to 39 respectively. Table 1 shows the concentrations of components constituting each sample in the medium in each well.

The plates were placed in a carbon dioxide incubator (5% CO ₂ , 37 ° C.) and PC12 cells were cultured for 72 hours.
(5) Evaluation of Cell Elongation After culturing in (4), the plate was spun to sediment cells in wells, and the supernatant was removed. Three microscopic images (bright field, × 10 images, 3 wells) were taken of PC12 cells which were precipitated in each well. The shape of all cells of PC12 cells shown in the image was observed. As shown in FIG. 1, in the PC12 cell, as the differentiation progresses, the process 2 extends from the cell body 1. The total number of PC12 cells and the number of cells having a protrusion 2 with a length L2 longer than the major axis L1 of the cell body 1 were counted. The number of cells having a protrusion having a length L2 longer than the major diameter L1 was divided by the total cell number to obtain an extension ratio in which the PC12 cells were extended. At least 80 or more PC12 cells appeared in each image. The total number of PC12 cells in all images was over 720. The expansion rate is shown as the average value of all PC12 cells in all images. The extension ratio for each PC12 cell is shown in FIG. In FIG. 2, “M”, “C”, “D” and “I” mean “6-MSITC”, “curcumin”, “DHA” and “isorhamnetin” in this order. The same applies to FIGS. 3 to 6. In FIG. 2, τ means that it is statistically significant. The symbol * means that a synergistic effect was recognized by combining two components. Here, the synergetic effect means that the extension ratio is larger when the two components are used in combination rather than the sum of the extension ratios when the respective components are used alone.

  As shown in FIG. 2, when any of 6-MSITC, curcumin, DHA, and isorhamnetin is contained in the medium, the cell elongation effect is enhanced as compared with the case where these are not contained. The cell elongation effect of 6-MSITC was higher than curcumin, DHA and isorhamnetin.

  When 6-MSITC was added to the medium, significant elongation acceleration was observed for PC12 cells when 6-MSITC was 0.1 μM, 0.5 μM, and 2.5 μM. When curcumin was added to the medium, significant elongation promotion was observed for PC12 cells when curcumin was 0.1 μM, 0.5 μM, and 2.5 μM. When DHA was added to the medium, significant elongation promotion was observed for PC12 cells when DHA was 0.5 μM, 2.5 μM, and 40 μM. When isorhamnetin was added to the medium, significant elongation promotion was observed for PC12 cells when isorhamnetin was 0.02 μM, 0.1 μM, and 0.5 μM.

  By combining two types of 6-MSITC and the other components, the elongation ratio was higher than when one type of component was used. In the case of the combination of 6-MSITC and curcumin, the elongation rate was effectively increased when 6-MSITC was 0.1 to 2.5 μM and curcumin was 0.1 to 2.5 μM. In the case of the combination of 6-MSITC and DHA, the elongation rate was effectively increased when 6-MSITC was 0.1 to 2.5 μM and DHA was 0.5 to 40 μM. In the case of the combination of 6-MSITC and isorhamnetin, the elongation rate was effectively increased when 6-MSITC was 0.1 μM, 2.5 μM and isorhamnetin was 0.02 to 0.5 μM.

(Experimental example 2)
The toxicity of each component to PC12 cells was investigated using MTT (3- (4,5-di-methylthiazol-2-yl) -2,5-diphenyltetrazolium bromide, yellow tetrazole) reagent.
In order to investigate the toxicity of each component to PC12 cells, PC12 cells were cultured in the same manner as (1) to (4) of the above (Experimental Example 1) except for the components to be added to the medium. The component added to the differentiation medium B in each well in (4) is either 6-MSITC (component 1), curcumin (component 3), DHA (component 2), or isorhamnetin (component 4). The concentration of each component in the differentiation medium B is 6 (for control), 0 (control), 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, 80 μM, 160 μM for 6-MSITC (component 1), and curcumin (component 3) Is 0 (control), 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, and DHA (component 2) is 0 (control), 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, 80 μM, 160 μM, The isorhamnetin (component 4) was 0 (control), 2.5 μM, 5 μM, 10 μM, 20 μM.

After culturing PC12 cells for 72 hours in (4) PC12 cell differentiation of the above (Experimental Example 1), 10 μl of MTT reagent (manufactured by Nacalai Tesque, product number 23547-21, MTT 5 mg / ml in phosphate buffered saline) / Well added. Furthermore, PC12 cells were cultured for 4 hours in a carbon dioxide incubator (5% CO ₂ , 37 ° C.).

  The plates were spun to sediment the cells in the wells and the supernatant removed. 100 μl / well of DMSO was added to each well, shaken and agitated, and PC12 cells were suspended in DMSO to obtain a cell solution.

  The absorbance at 490 nm was measured for the cell fluid in each well. The value obtained by converting the absorbance of the control cell fluid into a fraction using the absorbance of the cell fluid of the PC12 cells cultured in a culture solution containing each sample as a numerator and a fraction was calculated as a MTT assay. The measurement results in the case of using 6-MSITC, curcumin, DHA, and isorhamnetin are sequentially shown in FIG. 3, FIG. 4, FIG. 5, and FIG.

  When 6-MSITC, curcumin, DHA, and isorhamnetin were used, the MTT assay was higher than when they were not used (control). From this, it was found that 6-MSITC, curcumin, DHA, and isorhamnetin are components with low toxicity.

Claims (13)

6-methylsulfinylhexyl isothiocyanates or a glycoside thereof;
At least one member selected from the group consisting of unsaturated fatty acids and polyphenols;
Nerve cell elongation promoter.   The nerve cell elongation promoter according to claim 1, wherein the unsaturated fatty acid is an ω3 fatty acid.   The nerve cell elongation promoting agent according to claim 2, wherein the omega-3 fatty acid comprises one or more selected from the group consisting of docosahexaenoic acid and eicosapentaenoic acid.   The nerve cell elongation promoting agent according to claim 2 or 3, wherein the omega-3 fatty acid is extracted from fish and shellfish and / or fish oil.   The nerve cell elongation promoting agent according to claim 1, wherein the polyphenol comprises one or more selected from the group of curcuminoids and flavonoids.   The nerve cell extension promoter according to claim 5, wherein the curcuminoid comprises curcumin.   The nerve cell elongation promoter according to claim 5 or 6, wherein the curcuminoid is extracted from turmeric.   The nerve cell extension promoter according to claim 5, wherein the flavonoid comprises isorhamnetin.   The nerve cell extension promoter according to claim 5 or 8, wherein the flavonoid is extracted from Ginkgo biloba leaves.   The nerve cell elongation promoter according to claim 1, comprising 6-methylsulfinylhexyl isothiocyanates and docosahexaenoic acid.   The nerve cell elongation promoter according to claim 1, comprising 6-methylsulfinylhexyl isothiocyanates and curcumin.   The nerve cell elongation promoting agent according to claim 1, comprising 6-methylsulfinylhexyl isothiocyanates and isorhamnetin. Isothiocyanates,
At least one selected from the group consisting of unsaturated fatty acids and polyphenols;
Nerve cell elongation promoter.