KR101047590B1 - Oleanane Triterpene Saponin Compounds for the Treatment of Dementia and Mild Cognitive Impairment and Improving Cognitive Function - Google Patents

Abstract

Since the present invention has an effect of improving the memory and learning ability of the oleonan-based triterpene saponin compound is reduced, as an active ingredient of a health food that improves the brain function, such as treatments and cognitive function of diseases related to dementia and mild cognitive impairment It relates to the use of the oleonan-based triterpene saponin compound.

Cognitive function, memory, learning ability, oleanolic acid saponin, hederagenin saponin

Description

Oleanane triterpene saponin compounds which are effective on treatment of dementia and mild cognitive impairment, and improvement of cognitive function}

FIG. 1 is a graph showing the degree of memory-improving effect by scopolamine by oral administration of the olean-based triterpene saponin compound of the present invention once.

The present invention has the effect of improving the memory and learning ability of the oleonan-based triterpene saponin compound represented by the following formula (1), thereby improving brain functions such as treatments and cognitive functions for diseases related to dementia and mild cognitive impairment The present invention relates to the use of an oleonan-based triterpene saponin compound as an active ingredient of a health food.

In Formula 1, R ₁ , R ₂ and R ₃ each represent a hydrogen atom or an alkyl group of C ₁ -C ₄ , and R ₄ represents an alkyl group of C ₁ -C ₄ or a hydroxyalkyl group of C ₁ -C ₄ . , R ₅ and R ₆ are each hydrogen or sugar, and at least one of R ₅ and R ₆ represents sugar, wherein sugar is glucose, galactose, rhamnose, gyros , Arabinose, and glucuronic acid.

Triterpene saponin (sapogenin, aglycone) has a triterpene, and refers to a compound that forms a glycoside bond or an ester bond with sugar. Triterpene saponins are classified according to the type of saponenin, pentacyclic triterpenes such as oleanane, ursane, lupane, hopane, and taraxerane. Saponin which has as a saponenin is typical. In addition, saponins are distinguished according to the binding position with sugar chains. For example, oleanoic acid, which binds with a sugar chain at any one of the C-3 and C-28 positions, is called monodesmoside, and C-3 and C- 28 The position of bisdesmoside is distinguished from the sugar chain in two places. In the C-3 position, the sugar chain is mainly bonded by glycosidic bond, and in the C-28 position, the sugar chain is mainly bonded by ester bond. Sugars that primarily bind saponins are glucose, galactose, rhamnose, gyros, arabinose, glucuronic acid, and the like.

Saponins are known to have very low intestinal absorption in the bisdesmoside state. However, saponin is easily hydrolyzed by the enzyme of intestinal bacteria at the time of taking so that the sugar chain is removed and the sugar chain connected by the glucosidic bond at the C-3 position is partially hydrolyzed sequentially from the end. Absorbed by Kim DH, Bae EA, Han MJ, Park HJ, Choi JW. Metabolism of Kalopanax saponin K by human intestinal bacteria and antirheumatoid arthritis activity of their metabolites. Biol Pharm Bull . 2002 Jan; 25 (1) : 68-71.]. On the other hand, triterpene sapogenin itself, which has no sugar binding, has not only low solubility but also low intestinal absorption rate, which has been reported to be less active than glycosides in several animal experiments. [Yoshikawa M, Matsuda H. Antidiabetogenic activity of oleanolic acid glycosides from medicinal foodstuffs. Biofactors . 2000; 13 (1-4) : 231-7].

As the elderly population increases not only in Korea but also in the world, many degenerative diseases of the elderly are causing social and economic losses. Statistics released by the American Dementia Association and the National Institute of Aging indicate that 4 million Americans have dementia, typically dementia after age 60, but rarely start in their 50s and 10.3% of all Americans after age 65 The cost of treating dementia is $ 95 billion annually in the United States.

In Korea, the report of the Korea Institute for Health and Social Affairs showed that the dementia population increased rapidly due to aging, and the prevalence of dementia in 1995 was 8.3% among elderly people over 65 years old, and it is estimated to increase by 0.7 percentage point to 9% in 2020. In addition, as a result of applying the prevalence of dementia to the prospective population revealed by the National Statistical Office, the number of elderly people with dementia in 2000 was 277,00048 (8.3% of the elderly aged 65 and over), 52,6868 people (9%) in 2015. It is estimated that it will reach 61,132 (9%) in 2020. Dementia disease is a serious incurable disease that must be treated because it is a serious social and economic problem because it not only destroys the patient's own human life but also destroys the family life.

Dementia disease can be caused by a variety of causes, but the main symptom is a marked loss of learning and memory.

Tacrine, released in 1993 as the first FDA-approved drug to treat dementia, inhibits the breakdown of acetylcholine (ACh) produced in the brains of Alzheimer's disease patients in early and mid-stage by approximately 30% of patients. Loss of cognitive function could be delayed, but it is rarely used at present because it causes many side effects related to liver. Aricept, approved by the US FDA in 1996, works by increasing the use of acetylcholine, which can be taken once a day before bedtime. Side effects include nausea, diarrhea, and tiredness. Will disappear soon. Therefore, there is an urgent need to develop new dementia treatments with fewer side effects and good efficacy.

Accordingly, the present inventors have studied to develop a brain function improver that is effective in improving the ability to learn as well as the reduced memory as well as reduced toxicity, as a result of the treatment of oleanic triterpene saponin compounds with dementia and mild cognitive impairment and The present invention has been completed by knowing that the cognitive function is improved.

Accordingly, an object of the present invention is to provide a medicament and health food for improving brain function containing an olean-based triterpene saponin compound as an active ingredient.

The present invention is characterized by a medicament for treating dementia and mild cognitive impairment or a health food for improving brain function, which contains an oleanane-based triterpene saponin compound represented by the following formula (1).

[Formula 1]

In Formula 1, R ₁ , R ₂ and R ₃ each represent a hydrogen atom or an alkyl group of C ₁ -C ₄ , and R ₄ represents an alkyl group of C ₁ -C ₄ or a hydroxyalkyl group of C ₁ -C ₄ . , R ₅ and R ₆ are each hydrogen or sugar, and at least one of R ₅ and R ₆ represents sugar, wherein sugar is glucose, galactose, rhamnose, gyros , Arabinose, and glucuronic acid.

Hereinafter, the present invention will be described in detail.

Represented as the oleanane-based triterpene saponin compound represented by the formula (1) according to the present invention, oleanolic acid saponin represented by the following formula (1a) and hederagenin saponin represented by the following formula (1b) are already well known compounds in the art .

The oleanolic acid saponin represented by Chemical Formula 1a is a compound having oleanolic acid as sapongenin, and is anticancer and anti-inflammatory [Li J, Guo WJ, Yang QY. Effects of ursolic acid and oleanolic acid on human colon carcinoma cell line HCT15. World J Gastroenterol . 2002 Jun; 8 (3) : 493-5], Cerebral nerve protection [Qian YH, Liu Y, Hu HT, Ren HM, Chen XL, Xu JH. The effects of the total saponin of Dipsacus asperoides on the damage of cultured neurons induced by β-amyloid protein 25-35], an antivirus [Kapil A, Sharma S. Effect of oleanolic acid on complement in adjuvant- and carrageenan-induced inflammation in rats. J. Pharm Pharmacol. 1995 Jul; 47 (7) : 585-7], antihyperlipidemia [Lee KT, Sohn IC, Kim DH, Choi JW, Kwon SH, Park HJ. Hypoglycemic and hypolipidemic effects of tectorigenin and kaikasaponin III in the streptozotocin-lnduced diabetic rat and their antioxidant activity in vitro. Arch Pharm Res . 2000 Oct; 23 (5 ): 461-6], antiallergic [Park KH, Park J, Koh D, Lim Y. Effect of saikosaponin-A, a triterpenoid glycoside, isolated from Bupleurum falcatum on experimental allergic asthma. Phytother Res. 2002 Jun; 16 (4) : 359-63], immunomodulatory [Ju DW, Zheng QY, Cao X, Fang J, Wang HB. Esculentoside A inhibits tumor necrosis factor, interleukin-1, and interleukin-6 production induced by lipopolysaccharide in mice. Pharmacology . 1998 Apr; 56 (4) : 187-95], the efficacy of inhibiting angiogenesis [Korean Patent Registration No. 101,480] has already been known.

Hederagen saponin represented by Formula 1b is a compound having hederagenin as sapogenin, and has anti-inflammatory properties [Kwak WJ, Han CK, Chang HW, Kim HP, Kang SS, Son KH. Loniceroside C, an antiinflammatory saponin from Lonicera japonica. Chem Pharm Bull (Tokyo) . 2003 Mar; 51 (3) : 333-5], analgesic [Choi J, Huh K, Kim SH, Lee KT, Park HJ, Han YN. Antinociceptive and anti-rheumatoidal effects of Kalopanax pictus extract and its saponin components in experimental animals. J Ethnopharmacol . 2002 Feb; 79 (2) : 199-204], antioxidant [Choi J, Huh K, Kim SH, Lee KT, Lee HK, Park HJ. Kalopanax saponin A from Kalopanax pictus, a potent antioxidant in the rheumatoidal rat treated with Freund's complete adjuvant reagent. J Ethnopharmacol . 2002 Jan; 79 (1) : 113-8], blood sugar suppression [Kim DH, Yu KW, Bae EA, Park HJ, Choi JW. Metabolism of Kalopanax saponin B and H by human intestinal bacteria and antidiabetic activity of their metabolites. Biol Pharm Bull . 1998 Apr; 21 (4) : 360-5] Efficacy is already known.

However, according to the results published to date, the treatment effect of dementia and mild cognitive impairment and brain function improvement of the oleanane-based triterpene saponin compound represented by the formula (1) including oleanolic acid saponin and hederagenin saponin, None known.

However, Japanese Patent Application Laid-Open No. 2000-247993 confirms that oleanolic acid has a high affinity for sigma receptors. vascular disease, old age disorder, Alzheimer's disease, Parkinson's disease, Huntington's disease, drug addiction, as you mention that the efficacy against such stresses, in vitro experiments with these diseases directly in vivo animal experiments, or nerve cells involved in No results were given.

Oleanane-based triterpene saponin compounds having oleanoic acid or hederagenin as sapongenin are monodesmoside compounds having sugars bonded at positions C-3 or C-28 or C-3 and C Bisdesmoside compounds may be present in which sugar is bound at the -28 position. By various combinations of these sugar chains, there are more than 150 saponin compounds as illustrated below [Gangsamsik, Triterpenoid Saponins, Seoul National University Press, 1996, Tradimed Natural Products Database].

Oleanolic acid saponins :

Achyranthoside C, Acutoside A-G, Akeboside Stj, Anemoside A, Araliasaponin XII-XVIII, Araloside ) D, Arvensoside A to B, Betavulgaroside IV to V, Bupreureum Chinese triterpene glycoside S1, Caraganoside A, Chicuse Chikusetsusaponin IB, IV, Cijiujianoside A1, C4, C3, D1, Clematichinenoside A, C, Clematis chinensis prosapogenin CP9, CP9a , CP7a, CP2b, Clemontanoside E, F, Crocus sativus oleanoic acid saponin, Elatoside B, D, Eleutheroside K, Fatsiaside A1 , Hederacolchiside E, He Hexaasaponin B, Huzhangoside A, B, C, Lablaboside A, Lucioside H, 6'-Methylmomodine I, Momordin I, IC , IIB, 3-O-α-L-arabinopyranosyloleanoic acid, 3-O-α-L-arabinopyranosyloleanoic acid-28-O-β-D-glucosyl (1 → 6) β-D- Glucoside, 3-O-α-L-rhamopyranosyl (1 → 2) -α-L-arabinopyranosyloleanoic acid-28-O-β-D-gyropyranosyl (1 → 6) -β-D- Glucopyranosyl ester, 3-O-β-D-galactopyranosyl (1 → 2) -β-D-glucuronopyranosyl oleanoic acid, 3-O-β-D-glucopyranosyl (1 → 3 ) -α-L-lamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanoic acid, 3-O-β-D-glucopyranosyl (1 → 3) -α-L-lamnopyranosyl ( 1 → 2) -α-L-arabinopyranosyl oleanolic acid-28-genthiobioside, 28-O-β-D-glucopyranosyl oleanoic acid, 3-O-β-D-glucopyranosyl oleanoic acid, 3- O-β-D-glucuronopyranosyl Reanolic acid, 3-O-β-D-ribopyranosyl (1 → 3) -α-L-ramnopyranosyl (1 → 2) -α-L-arabinofyranosyl oleanoic acid, 3-O-β-D Gyropyranosyl (1 → 3) -α-L-Rhamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanoic acid, 3-O-β-D-gyrosil (1 → 4) -β -D-glucosyl (1 → 4) -α-L-lamnosyl (1 → 3) -β-D-glucosyl (1 → 3) -α-L-lamnosyl (1 → 2) -α-aravies Nosyl oleanolic acid-28-O-β-D-glucosyl (1 → 6) β-D-glucosyl ester, 2'-0-glucopyranosyl momodine Ic, 2'-0-glucopyranosyl momodine Iic, 3-O-β-D-gyropyranosyl (1 → 4) -β-D-glucopyranosyl (1 → 3) -α-L-rhamnopyranosyl (1 → 2) -α-L-arabinofi Lanoside oleanolic acid-28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranosyl ester, oleanoic acid 3-O-neosperidoside, oleanoic acid-3-O-α-L -Arabinopyranosyl 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranosyl ester, oleanoic acid-3-O-β-D-glucose Lanosyl (1 → 2) -α-L-arabinopyranoside, Pericarpsaponin J3, Prosapogenin CP4, Quinatoside D, Radeanin A, C , D, E, F, Radedeanoside R10, R11, Riburinin (Rivularinin), Spinacoside C, D, Spinasaponin A, Tarasaponin II, III , VI, Tetrapanax papyriferum saponin R-3, R-1-a, R-4b, Udosaponin methyl esters A-C.

Hederagenin saponins :

Akevia saponins A to G, Akeboside, Asperosaponin F, H1, Calcoside D, Cauroside D, F, Clematis Chinese Prosapogenin (Clematis chinensis) prosapogenin) CP10, CP10a, Cp8a, CP3b, CP0, CP3a, CP2a, 3-O-β-D-gyropyranosyl (1 → 3) -α-L-arabinofyranosyl hederagenin-28-O-α -L-ramnopyranosyl (1 → 4) -β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside, hederagenin-3-O-α-L-arabinoside, Hederagenin-3-O- [α-L-lamnopyranosyl- (1 → 2) -α-L-arabinopyranosyl] -28-O-β-D-gyropyranosyl (1 → 6)- β-D-glucopyranosyl ester, hederagenin-3-O- [α-L-ramnopyranosyl (1 → 2) -α-L-arabinopyranosyl] -28-O- [3-O- Acetyl-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside, hederagenin-3-O- [α-L-rhamnopyranosyl (1 → 2) -α-L- Arabinopyranosyl] -28-O- [3-O-acetyl-β-D-gyropyranosyl (1 6) -β-D-glucopyranoside, hederagenin-3-O-α-L-rhamnopyranosyl (1 → 3) -β-D-glucopyranosyl (1 → 3) -α-L- Rampofyranosyl (1 → 2) -α-L-arabinopyranosyl 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranosyl ester, hederagenin-3 -O-β-D-glucopyranosyl (1 → 3) -α-L-lamnopyranosyl (1 → 2) -α-L-arabinopyranosyl 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranosyl ester, hederagenin-3-O- (4-O-acetyl) -α-L-arabinofyranosyl 28-O-β-D-glucopyranosyl (1 → 6) -β-D-glucopyranosyl ester, Kalopanaxsaponin B, G, JLa, JLb, Leontoside A, B, Luciyoside A, E, Mucologosaphonon ( Mukurozisaponin) EI, G, X, Y2, YI, 4'-O-acetylacecaviasaponin D, 3-O- [α-L-arabinosyl] hederagenin-28-O- [β-D-glucosyl ] Ester, 3-O-α-L-lamnopyranosyl (1 → 3) β-D-glucosyl (1 → 3) α-D-lamnosyl (1 → 2) α-ara Nosyl hederagenin-28-O-β-D-glucosyl (1 → 6) β-D-glucosyl ester, 3-O-β-D-glucopyranosyl (1 → 3) -α-L-rhamno Pyranosyl (1 → 2) -α-L-arabinopyranosyl hederagenine, 3-O-β-D-glucopyranosyl hederagenin, 3-O (β-D-glucosyl (1 → 4) α-L-Rhamnosyl (1 → 3) β-D-glucosyl (1 → 3) α-D-Rhamnosyl (1 → 2) α-arabinosyl hederagenin-28-O-β-D-glucose Sil (1 → 6) β-D-glucosyl ester, 3-O (β-D-glucosyl (1 → 4) α-L-lamnosyl (1 → 3) β-D-glucosyl (1 → 3) α-L-Rhamnosyl (1 → 2) α-arabinosyl hederagenine, 3-O (β-D-gyrosil (1 → 4) β-D-glucosyl (1 → 4) α-L- Rhamnosyl (1 → 3) β-D-glucosyl (1 → 3) α-L-arabinosyl (1 → 2) α-arabinosyl hederagenine, 3-O-[(2'-O-acetyl) -α-L-arabinopyranosyl (1 → 6) -β-D-glucopyranosyl] hederagenin, 3-O- (2'-O-acetyl) -α-L-arabinofyranosyl hedera Genine-28-O- [β-D-glucopyranosyl (1 → 6) -β-D-glucopyranoside], percapsaponin (Percarpsa ponin) C, J2, G, K, Pulsatiloside A, B, C, Quinatoside A, B, C, Sapindoside A, B, C, Staunoside (Staunoside) A, B, D, E, Tauroside G3, Udosaponin methyl esters D, E, F.

Oleanane-based triterpene saponin compounds, which typically include oleanolic acid saponins and hederagenin saponins, are extracted from the plants exemplified by the following general extraction methods: Amaranthaceae plants, for example Achyranthes sp. , amaranth (Amaranthus sp.), etc., Araliaceae (Araliaceae), for plants for senticosus (Acanthopanax sp.), aralia (Aralia sp.), Fatsia Japonica (Fatsia sp.), kalopanax (kalopanax sp.), ginseng (Panax sp .), Bottled talmok (Tetrapanax sp.), etc., Basel, etc. rasegwa (Basellaceae) overseen bosing, for plants for example thiazole (Boussingaultia sp.), berberidaceae (Berberidaceae) plants, for example in the pheasant leg Buttercup (Caulophyllum sp.), etc. , Boraginaceae plants such as Anchusa sp., Caprifoliaceae plants such as Lonicera sp., Etc. Chenopodiaceae plants such as Chenopodium sp. Back, gourd ( Cucu rbitaceae plants such as Actinostemma sp., Luffa sp., Momordica sp., etc. Dipsacaceae plants such as Dipsacus sp. Euphorbiaceae plants, such as Putranjiva sp., Hippocastanaceae plants, such as Aesculus sp., Etc., Lardizabalaceae plants, such as Akebia sp. Leguminosae plants, for example Acacia sp., Albizzia sp., Swartzia sp., Etc. Opiliaceae plants, for example Opilia sp. .) Phytolaccaceae plants, such as Phytolacca sp., Etc., Ranunculaceae plants, such as Anemone sp., Clematis sp., Hedera sp. .), Pulsatilla sp., Etc. Rubiaceae plants, for example Randia dia sp.), serom piece (Xeromphis sp.), etc., sapindaceae (Sapindaceae) Pomeranian thiazol example plants example (Pometia sp.), spin Ranges (Spindus sp.), Tino Wu ah (Thinouia sp.), etc., valerianaceae Valerianaceae plant, for example Patrinia sp.

According to the results of animal experiments by the inventors of the present invention, scopolamine (scopolamine) and a drug-free control (control), scoflavin (1 mg / kg) known to reduce memory through the interruption of the transmission of neurotransmitters As a result of evaluating memory enhancement between the group administered with and the group administered with the oleanane-based triterpene saponin compound represented by Formula 1 1 hour after the administration of scoflavin, a significant activity was shown.

Therefore, the oleanane-based triterpene saponin compound represented by Formula 1 may be used as an active ingredient of a medicament for treating dementia and mild cognitive disorders and a health food for improving cognitive function.

In the case of a medicament containing an olean-based triterpene saponin compound represented by Chemical Formula 1 as an active ingredient, it may be provided in the form of a general pharmaceutical formulation that can be administered orally or parenterally during clinical administration. In the formulation process for oral or parenteral administration, pharmaceutically or food-acceptable fillers, extenders, binders, wetting agents, diluents or excipients such as disintegrants and surfactants can be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which solid preparations comprise at least one excipient such as starch, calcium carbonate, sucrose or the like in lignan and lactone compounds and derivatives thereof. It is prepared by mixing lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral administration include suspensions, solvents, emulsions, and syrups. In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have.

Formulations for parenteral administration include sterile aqueous solutions, water-insoluble solvents, suspensions, emulsions, lyophilizers, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, utopsol, macrogol, tween 61, cacao butter, laurin butter, glycerol gelatin and the like can be used.

The content of the active ingredient in the drug according to the present invention can be appropriately selected depending on the absorbency, inactivation rate, excretion rate, age, gender and condition of the active ingredient in the body. The daily dosage based on the olanan-based triterpene saponin compound represented by the formula (1) is 0.1 to 10 mg / kg, preferably 0.5 to 5 mg / kg, and may be administered 1 to 3 times a day. .

In addition, the olanan-based triterpene saponin compound represented by Formula 1 may be provided in the form of a health food containing it as an active ingredient. The health food in the present invention is a food prepared by adding the olanan-based triterpene saponin compound represented by the formula (1) to general foods, or by encapsulating, powdering, suspensions, and the like. Means to bring the effect, unlike the general medicine has the advantage that there is no side effect that can occur when using the drug as a raw material for long-term use.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1 Oleanan Triterpene Saponin Extraction

The oleanolic acid saponin and hederagenine saponin were isolated from the following herbal medicines, which have been reported to contain a large amount of oleanolic acid saponin and hederagenin saponin in the existing literature.

Arlaia elata , Pulsatilla chinensis , Lonicera japonica and Kalopanax pictus were taken at 1 kg dry weight and refluxed in 7 L of 50% ethanol for 4 hours. It was carried out times. The extract was filtered and concentrated under reduced pressure at 50 ° C. with a rotary evaporator. Suspended to about 5 times the original crude weight (V / W) by adding water to the resulting concentrate, followed by equal saturation. The mixture was mixed with normal-butanol, placed in a separation funnel, and left for 24 hours after stirring to separate only the upper butanol layer. The fractions were re-distilled two to three times more, and the butanol fractions were concentrated using a distiller and then dried by removing the solvent completely in a vacuum oven.

Column chromatography was performed on the normal butanol fraction with octadecylsilylated silica resin (YMC * GEL ODS-A 12 nm, S-150 m). The amount of resin used was 250 g, which is 25 times the amount of 10 g of the sample, and the solvent was 2-3 times the volume of the resin by increasing the amount of methanol by 10% (V / V) from 10% (v / v) aqueous methanol solution. Was used as a step-gradient method. Only fractions separated by 70%, 80%, 90% aqueous methanol solution (V / V) were taken to maximize concentration of the saponin compound.

Example 2: Single Component Separation and Structural Analysis of Oleanan Triterpene Saponin

High performance liquid chromatography (HPLC) using acetonitrile and water as a mixed solvent of methanol aqueous solution fractions and analysis using a PDA detector have a maximum absorbance at 210 nm and are specific to triterpenes. The main peak showing the spectrum was selected. Subsequently, the selected peaks were separated purely using Preparative HPLC using a mixed solvent of 9.5 ml of acetonitrile and water per minute, concentrated using a distillation apparatus, and dried by completely removing the solvent in a vacuum oven. The column used was YMC J'Sphere ODS-H80, and the wavelength was measured at 210 nm. In addition, sugars were removed from saponins by heating the components of the separated peaks at 100 ° C. for 60 minutes under acidic conditions of 2N H ₂ SO ₄ . HPLC analysis of the hydrolyzate with oleanolic acid and hederagenine standard purchased from Sigma proceeded to further component separation for the component identified as oleanolic acid saponin or hederagenine saponin. Eight single materials such as 1 were obtained.

¹ H-NMR (500 MHz), ¹³ C-NMR (125 MHz), DEPT, 2D NMR ( ¹ H- ¹ H COZY, HMQC, HMBC, TOCSY, NOESY) were measured to determine the structure of the isolated single material. Based on this data, structural decisions were made. Acid hydrolyzed sugars were carbohydrate analysis system and TMS derivatization and analyzed by GC to confirm the type and number of sugars. Structural analysis of each sugar was performed by 2D NMR ( ¹ H- ¹ H COSY, HMQC, HMBC, TOCSY), and the linkage of sugar was performed by 2D NMR (HMBC, NOESY).

Using the above method, 100 to 500 mg of eight oleanolic acid saponins or hederagen saponins as shown in Table 1 were obtained.

division Scientific name Herbal medicine Molecular formula Molecular Weight Ingredient ① Eleutheroside K
(3-O-α-L-ramnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanoic acid) Baekduong
( Pulsatilla
chinensis ) C ₄₁ H ₆₆ O ₁₁ 735.0 Ingredient ② Hederasaponin B
(3-O-α-L-lamnopyranosyl (1 → 2) -α-L-arabinopyranosyl oleanoic acid-28-O-α-L-lamnopyranosyl (1 → 4) -β-D-glucose Pyranosyl (1 → 6) -β-D-glucopyranosyl ester) Baekduong
( Pulsatilla
chinensis ) C ₅₉ H ₉₆ O ₂₅ 1205.4 Ingredient ③ Hedera colchiside E
(3-O-α-L-lamnopyranosyl (1 → 2) -β-D-glucopyranosyl (1 → 4) -α-L-arabinopyranosyl oleanoic acid-28-O-α-L-rhamno Pyranosyl (1 → 4) -β-D-glucopyranosyl (1 → 6) -β-D-glucopyranosyl ester) Baekduong
( Pulsatilla
chinensis ) C ₆₅ H ₁₀₆ O ₃₀ 1366.6 Ingredient ④ Elatoside A
(3-O-β-D-gyropyranosyl (1 → 2) -β-D-galactopyranosyl (1 → 3) -β-D-glucuronopyranosyl oleanoic acid) Neck
( Aralia
elata ) C ₄₇ H ₇₄ O ₁₈ 927.1 Ingredient ⑤ Elatoside C
(3-O-β-D-gyropyranosyl (1 → 2) -β-D-galactopyranosyl (1 → 3) -β-D-glucuronopyranosyl oleanoic acid-28-O-β- D-glucopyranosyl ester) Neck
( Aralia
elata ) C ₅₃ H ₈₄ O ₂₃ 1089.2 Ingredient ⑥ Loniceroside A
((3-O-α-L-arabinopyranosyl hederagenin-28-O-α-L-rhamnopyranosyl (1 → 2) -β-D-gyropyranosyl (1 → 6) -β- D-glucopyranosyl ester) Lighting
( Lonicera
japonica ) C ₅₂ H ₈₄ O ₂₁ 1045.2 Ingredient ⑦ Loniceroside B
(3-O-α-L-rhamnopyranosyl (1 → 2) -α-L-arabinopyranosyl hederagenin-28-O-α-L-rhamnopyranosyl (1 → 2) -β-D Gyropyranosyl (1 → 6) -β-D-glucopyranosyl ester) Lighting
( Lonicera
japonica ) C ₅₈ H ₉₄ O ₂₅ 1191.4 Ingredient ⑧ Kalopanaxsaponin B
(Pericarpsaponin Pk, 3-O-α-L-lamnopyranosyl (1 → 2) -α-L-arabinopyranosyl hederagenin-28-O-α-L-ramnopyranosyl (1 → 4)- β-D-glucopyranosyl (1 → 6) -β-D-glucopyranosyl ester) Thawed blood
( Kalopanax
pictus ) C ₅₉ H ₉₆ O ₂₆ 1221.4

Example 3: Passive avoidance test for triterpene saponins

A passive avoidance test was conducted to determine whether the eight oleanolic acid saponins or hederagenin saponins were effective in improving memory, a major symptom of dementia in vivo. The aglycons of these saponin compounds, oleanolic acid and hederagenin, were also included in the experiment.

The experimental apparatus used a shuttle box having a width, length, and height of 50, 15, and 40 cm. The box is divided into two rooms using a guillotine door, one of which is illuminated, which makes it a bright room, and the other is covered by a black cloth, making it a dark room. To make it possible.

First, if you put the mouse in a bright room, turn on the lights and open the partition door, the mouse will enter the dark room within 20 seconds according to the instinct to find a dark place, and the partition door will close as soon as the mouse moves to the dark room. In this way, the time required for the rats to enter the dark room from the light room is measured as the latency time, and a training trial was conducted on the first day of the experiment to make sure that all the mice fit within 20 seconds.

The next day, put the above trained rats back in the bright room and turn them on in the dark room. At this point, an electric stimulus of 0.8 mA is applied for three seconds through an electronic grid installed on the floor of a dark room, causing the mice to be shocked at the soles of their feet.

After 24 hours of this acquisition trial, rats were placed in a bright room, turned on, and led to a dark room. Normal rats remembered the shock of the previous day and hesitated to enter the dark room. The arrival time at this time was measured again with a maximum of 300 seconds.

In the experiment, one hour after the administration of scopolamine and scopolamine (1 mg / kg), which is known to reduce memory through interference with neurotransmitters, the control group without scopolamine and drugs was administered. mg / kg), aglycone (30 mg / kg), aricept (donepezil, 1 mg / kg) as a positive control, water retention as a negative control group, memory retention effect of the passive avoidance test (Rention time) Measured by.

As a result, the oral administration of the control drug Aricept showed a 1.6-fold increase in retention time compared to the negative administration of the water-treated group, and eight oleanoic acid saponins according to the present invention. Or, all of the components of hederagenin saponin are slightly different depending on the composition of aglycon and sugar, but all of them have a cognitive improvement effect of 2.5 to 3.0 times longer retention time in the passive avoidance test compared to the negative control group. Seemed.

These saponins also showed higher cognitive function than aglycon oleanolic acid (2.4 times) and hederagenin (2.3 times). TABLE 2, see FIG. 1.

These results suggest that oleanolic acid saponin and hederagenin saponin have the effect of improving the decreased memory in symptoms such as dementia disease and mild cognitive impairment. It was also found to be superior to oleanolic acid and hederagenin, which are aglycons.

process Residence time (seconds)  normal 286.9 ± 5.8  water  61.5 ± 22.4  Oleanolic acid  149.4 ± 19.7  Hederagenin  142.8 ± 32.6  Aricept  98.5 ± 10.6 Oleanolic acid saponins  Eleutheroside K  182.6 ± 15.9  Hederasaponin B  172.3 ± 29.1  Hedera colchiside E  155.7 ± 31.2  Elatoside A  166.0 ± 24.8  Elatoside C  160.1 ± 15.6 Hederagenin saponin  Loniceroside A  175.3 ± 21.1  Loniceroside B  169.7 ± 25.4  Kalopanaxsaponin B  170.5 ± 24.6

In the following formulation example, as an example for the manufacture of a medicament or health food comprising the oleanane-based triterpene saponin compound represented by the formula (1) as an active ingredient, the present invention is not limited to this type of formulation.

Preparation Example 1 Preparation of Powder and Capsule

A powder was prepared by mixing 50 mg of an oleonan-based triterpene saponin compound together with 74 mg of lactose, 15 mg of crystalline cellulose, and 1 mg of magnesium stearate. Furthermore, this powder was filled into No. 5 gelatin capsules using a suitable apparatus to prepare a capsule.

Preparation Example 2 Preparation of Liquid

50 mg of oleanane-based triterpene saponin compound was added to 20 g of sugar, 20 g of isomerized sugar, and lemon flavor, and adjusted to a total volume of 100 mL by adding sterile purified water, and then filled into a brown bottle and sterilized to prepare a liquid.

Preparation Example 3 Preparation of Health Food

On a daily basis, 100 mg of oleanane-based triterpene saponin compounds represented by Formula 1, ginseng extract 100 mg, green tea extract 100 mg, vitamin C 100 mg, powdered vitamin E 120 mg, iron lactate 2 mg, zinc oxide 2 mg, nicotinic acid amide 20 mg, vitamin A 5 mg, vitamin B1 2 mg, vitamin B2 2 mg, corn starch 200 mg, and magnesium stearate 20 mg were mixed.

As described above, the oleanane-based triterpene saponin compound represented by Chemical Formula 1 has excellent memory enhancement effect, and thus is a health food for improving brain function such as therapeutic drugs and cognitive function for diseases related to dementia and mild cognitive impairment. useful.

Claims (4)

A pharmaceutical composition for the treatment and prevention of dementia and mild cognitive impairment, comprising the oleonan-based triterpene saponin compound represented by the following Formula 1 as an active ingredient. [Formula 1]

In Formula 1, R ₁ , R ₂ and R ₃ each represent a hydrogen atom or an alkyl group of C ₁ -C ₄ , and R ₄ represents an alkyl group of C ₁ -C ₄ or a hydroxyalkyl group of C ₁ -C ₄ . , R ₅ and R ₆ are sugars selected from glucose, galactose, rhamnose, gyros, arabinose, and glucuronic acid, respectively. The method of claim 1, wherein the compound represented by Formula 1 is Hederasaponin B, Hedera colchiside E, Eratoside C, Loniceroside A, Lonieroside (Loniceroside) B, and Karopanaxsaponin B is a compound selected from the pharmaceutical composition. delete delete

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