KR20110081135A - Health food and therapeutic agent containing flavonoid compounds extracted from silk of zea mays linne for improving urinary system disorder - Google Patents

Abstract

PURPOSE: A therapeutic agent and a health food for treating urinary system disorders are provided to remarkably suppress bladder smooth muscle cells and prostate smooth cells. CONSTITUTION: A therapeutic agent and a heath food for treating urinary system disorder containing flavonoid compounds of chemical formula 1 isolated from Maydis stigmata. The isolated flavonoid compounds are chrysoeriol, chrysoeriol-6-C-beta boivinopyranosyl-7-O-beta-glucopyrnoside, and luteolin.

Description

Health food and therapeutic agent containing flavonoid compounds extracted from silk of Zea mays linne for improving urinary system disorder}

The present invention relates to a dietary supplement for treating urinary tract disease symptoms and a therapeutic agent for urinary tract diseases containing the flavonoid compound represented by the following formula (1).

<Formula 1>

Representative disease of urination disorder is prostate hyperplasia. Prostatic hyperplasia is found in 50% of elderly people in their 60s, and the main condition is hyperproliferation and hyperplasia of prostate epithelial and interstitial tissue. Factors that control prostate cell proliferation and death include endocrine factors, growth factors, interstitial and epithelial cell interactions, neuroendocrine and neurotransmitters, cell cycle and death control agents, Inflammation is mentioned.

Another major part of determining the condition of prostatic hyperplasia is secondary changes in the bladder due to impaired urine output. The obstruction of the bladder outlet causes hypertrophy of the bladder muscle, which causes irritable bladder due to excessive activity of the detrusor muscle, and if the obstruction persists for a long time, the contractile force of the detrusor muscle becomes inadequate, resulting in an increase in residual urine volume and urinary tract, resulting in urinary tract infections, Complications such as bladder stones and kidney failure occur.

In addition, the state causing bladder irritation symptoms are referred to as an overactive bladder, lower urinary tract symptoms, etc., including prostate hyperplasia in the lower urinary tract symptoms.

Overactive bladder (OAB) is a condition that is mainly caused by urinary urges that cause sudden urinary irrespective of the will, with or without urinary incontinence. Overactive bladder may be accompanied by an enlarged prostate, and the incidence of the disease is very high (15-20%) on average, and more than one-third of the elderly population, regardless of gender, are over 60 years old. Known. Most of these patients control urination by reducing fluid intake, but this also reduces external activity. This situation causes problems such as constipation in the elderly, and when a constipation can be repeated a vicious cycle that worsens the symptoms of urination again.

Lower urinary tract symptoms (LUTs) refer to symptoms of urinary storage and discharge, including prostate hyperplasia, bladder neck disorders, and neurogenic disorders. Since prostatic hyperplasia is blocked by the bladder outlet, it is difficult to discharge urine, so there are three types of problems according to the course of the disease. The first is that the bladder muscle takes more load than normal, so the bladder walls become thicker and more sensitive, causing frequent urination, nocturia, and residual urination. Second, if the bladder muscle becomes sensitive without fully draining the urine in the bladder, residual urine remains. Third is the type of urinary tract can come when the bladder is weakened and can not be normally discharged.

The bladder is primarily a sympathetic nerve stimulator that activates and contracts the muscarinic receptors in the bladder. In addition, the sympathetic nerve causes contraction through the a1 receptor in the bladder neck and prostate urethra, and the external urethral sphincter located in the mesothelial urethra may control the contraction relaxation by the body nerve. Urinary epithelium and nerve endings have the function of recognizing harmful stimuli by vanilloid receptors.

Currently, the most commonly used drug for the treatment of prostatic hyperplasia is alpha blocker. Alpha blockers (tamsulosin, doxazosin, alfuzosin, etc.) relax the bladder outlet by causing the prostate to develop well. However, about one-fifth of patients with prostatic hyperplasia lack an alpha-blocking agent alone, which prevents bladder hypersensitivity, that is, antimuscarinic agents (e.g., ditropan, tolterodine, Solifenacin (solifenacin, etc.) and antipurinergic agents (propiverine HCl), such as a combination administration, and will show a therapeutic effect.

On the other hand, Okmi-su is a graminse plant and is called Okmokseo's beard. It is also called Okmaeksu, Bonjamo, and Okmok calligraphy. Okmi water has been reported to have 2% fat, 0.12% volatile essential oil, 3.8% gum, 1.15% resin, 3.2% saponin, and 0.05% alkaloid.

Chrysoeriol and chryseriol-6-C-βvobinopyranosyl-7-O-β-glucopyranoside (Chrysoeriol-6-C-), which are flavonoid compounds from jade water β-boivinopyranosyl-7-O-β-glucopyranoside) and luteolin were extracted and isolated, and the compounds were reported to be active for the treatment of diabetes (Chem. Pharm. Bull., 2007, 55 (1)). , 153-155 and J. Nat. Prod., 2003, 66 (4), 564-565).

In addition, flavonoid compounds such as chryseriol and luteolin were isolated from the water-soluble ethanol extract of Artemisia vulgaris outpost, and the antioxidant effect of each of these isolated flavonoid compounds showed high activity compared to vitamin E. It has been reported (Korean Journal of Food Science and Technology, 1999, Vol. 31, No. 3, 815 ~ 822).

The inventors of the present invention are flavonoid-based compounds Chrysoeriol, Chryseriol-6-C-β boinopyranosyl-7-O-β-glucopyranoside (Chrysoeriol-6-C-β-boivinopyranosyl- 7-O-β-glucopyranoside) and Luteolin were extracted and separated from jade water to investigate the use of the flavonoid compound, and the flavonoid compound was investigated for the development of prostatic hyperplasia. By identifying the pathophysiology of the inhibitory effect and secondary changes of the bladder, the present invention can be usefully utilized as a dietary supplement for improving the symptoms of urinary tract diseases and the treatment of urinary tract diseases.

In the present invention, chrysoeriol and chryseriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside, which are flavonoid-based compounds extracted and separated from jade water (玉米 鬚) (Chrysoeriol-6-C-β-boivinopyranosyl-7-O-β-glucopyranoside), and nutraceuticals and therapeutic agents having new uses for Luteolin.

In order to achieve the above object, the present invention provides a dietary supplement for treating the symptoms of urinary tract diseases and the treatment of urinary tract diseases containing a flavonoid compound represented by the following formula (1).

<Formula 1>

Wherein R ₁ is hydrogen or C ₁ -C ₅ alkyl, R ₂ is hydrogen or β-boivinopyranosyl, R ₃ is hydrogen or β-glucopyranoside.

Flavonoid compounds represented by Formula 1 according to the present invention can be extracted and separated from jade water ( Zea mays linne ,玉米 鬚, corn beard) (Chem. Pharm. Bull., 2007, 55 (1), 153 ~ 155 And J. Nat. Prod., 2003, 66 (4), 564-565).

In the present invention, chryseryol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside (Chrysoeriol-6-C-β), which is a flavonoid-based compound from oxime water ethyl acetate (EtOAc) fraction -boivinopyranosyl-7-O-β-glucopyranoside), chrysoeriol and luteolin compounds were isolated.

Schematic diagram of isolating the flavonoid family of compounds from jade water according to the present invention is shown briefly as follows.

In the present invention, chrysoeriol and chrysoriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside (Chrysoeriol-6-C-β-boivinopyranosyl-7-O- β-glucopyranoside) showed inhibition of bladder contraction in the condition of contraction with a high concentration of K solution, which is a condition of calcium channel activation, and various K channel blockers (1 mM and / or 10 mM TEA, 10 μM 4-AP). Even when administered, it showed an effect of inhibiting bladder contraction, and the Luteolin compound inhibits the bladder contraction by blocking calcium channel at high concentrations, but at low concentrations, it may activate the Rho kinase pathway to cause abnormal contraction of the bladder. It has a dual effect and can be applied to an overactive bladder when administered at high concentrations.

In addition, the flavonoid-based compound according to the present invention significantly inhibits contraction caused by phenylephrine, and has an effect of relaxing the urethra like an alpha blocker commonly used in patients with prostatic hyperplasia.

Flavonoid compounds according to the present invention can be prepared by oral administration by mixing pharmaceutically acceptable conventional excipients, binders, disintegrants, lubricants, sweeteners and the like. At this time, as an excipient, microcrystalline cellulose, lactose, corn starch, mannitol, low-substituted hydroxypropyl cellulose, magnesium metasilicate aluminate, colloidal silicon dioxide, etc. are preferable, and as a binder, hydroxypropyl methyl cellulose, povidone, hydride, etc. Roxypropyl cellulose is preferred, and disintegrant is preferably croscarmellose sodium, sodium starch glycolate, calcium carboxymethyl cellulose, crospovidone and the like, and the lubricant is preferably magnesium stearate, talc, sodium stearyl fumarate, or the like. Do. The present invention is a preparation for oral administration, it is possible to manufacture pills, powders, granules, tablets, capsules, liquid formulations, and the like, can be prepared as a beverage or a tea, and the present invention is in addition to the active ingredient described above for administration In addition, it is prepared containing one or more pharmaceutically acceptable carriers, pharmaceutically acceptable carriers are saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol and these components One or more components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions and the like.

In addition, the dosage or dosage of the flavonoid compound according to the present invention varies depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the adult standard It is preferable to take a dose of 0.01 mg / kg to 1000 mg / kg divided once to several times a day.

Chryseriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside (Chrysoeriol-6-C-β-boivinopyranosyl-7-O-β-, which is a flavonoid compound according to the present invention) Glucopyranoside, Chrysoeriol and Luteolin compounds significantly inhibit the contraction of bladder smooth muscle and prostate smooth muscle by phenylephrine and acetylcholine, which have effects on urinary system diseases such as prostatic hypertrophy and overactive bladder As a health functional food for improving the symptoms of patients with urinary system diseases and can be used as a therapeutic agent for urinary system diseases.

Figure 1 shows the TLC pattern of the jade water ethyl acetate (EtOAc) fractionation layer,
Figure 1a shows the degree of color development in UV,
Figure 1b shows the degree of color development with 10% sulfuric acid solution,
Figure 1c shows the degree of color development with FeCl ₃ solution.
Figure 2 relates to a chryseryol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound according to the present invention isolated from the octaethylethyl acetate (EtOAc) fractionation layer,
2a shows a TLC pattern,
2b shows 1 H-NMR spectroscopy,
2C shows 13C-NMR spectroscopy,
2D shows Mass spectroscopy.
Figure 3 relates to a chryseriol compound according to the present invention isolated from the jade water ethyl acetate (EtOAc) fractionation layer,
3A shows a TLC pattern,
3b shows 1 H-NMR spectroscopy,
3C shows 13C-NMR spectroscopy,
3D shows Mass spectroscopy.
Figure 4 relates to a luteolin compound according to the present invention isolated from the jade water ethyl acetate (EtOAc) fractionation layer,
4A shows a TLC pattern,
4B shows 1 H-NMR spectroscopy,
4C shows 13C-NMR spectroscopy,
4D shows Mass spectroscopy.
FIG. 5A is a graph showing the effect of chrysriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound in PE on normal SD rat bladder,
FIG. 5B is a graph showing the effect of chrysriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound on ACh on normal SD rat bladder.
6a is a graph showing the effect of chryseriol compounds in PE on normal SD rat bladder,
6B is a graph showing the effect of chryseriol compounds on ACh on normal SD rat bladder.
7A is a graph showing the effect of luteolin compounds in PE on normal SD rat bladder,
7B is a graph showing the effect of luteolin compounds on ACh on normal SD rat bladder.
FIG. 8 is a graph showing the effect of chrysriol-6-C-β-boibinopyranosyl-7-O-β-glucopyranoside compound in PE on normal rabbit prostate and prostate urethral tissue.
9 is a graph showing the effect of chryseriol compounds in PE on normal rabbit prostate and prostate urethral tissue.
10 is a graph showing the effect of luteolin compounds in PE on normal rabbit prostate and prostate urethral tissue.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Experimental Examples.

< Example  1> Isolation and Identification of the Compounds According to the Invention from Corn Beard Extract

1) sub-fraction extraction

Filtration of hot water extracted from hot water (658.82 g) was filtered under reduced pressure to obtain 627.54 g of an aqueous part. The aqueous part was subjected to solvent fractionation with ethyl acetate (EA) to obtain 62.83 g of an ethyl acetate fraction. Ethyl acetate fractions were added from 30% methanol to 10% gradient to raise the concentration to 100% methanol , and then separated into 7 sub-fractions by low pressure liquid column chromatography, and TLC (developing solvent was chloroform: methanol: Water = 80: 20: 2) Various components were distributed in 3 and 4 sub-fractions by UV, 10% sulfuric acid, and FeCl ₃ , respectively, to confirm color development (FIGS. 1A to 1C).

2) Separation and purification of chrysriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound according to the present invention

The third sub-fraction (3.07g) was added from 60% methanol to 10% gradient to increase the concentration to 100% methanol, Sephadex LH-20 column chromatography was performed, and the resulting solution was concentrated. A major spot obtained by performing MCI gel CHP 20P while increasing the concentration to 100% methanol by 10% gradient was prep-TLC (chloroform: methanol: water = 75: 25: 3) to carry out a crystal of the following Chemical Formula 2 24 mg of all-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound was isolated and purified.

<Formula 2>

It was confirmed that it was separated into a single compound through TLC patterns and is shown in Figure 2a, 1H, 13C-NMR spectroscopy and mass spectroscopy were carried out and analyzed as shown in Figures 2b to 2d.

3) Separation and purification of chryseriol and luteolin compounds according to the present invention

The fourth sub-fraction (3.08g) was added to 10% gradient from water to increase the concentration to 100% methanol, and the solution obtained after YMC-gel ODS-A was concentrated, and the concentrate was concentrated from 100% ethanol to 10%. Sephadex LH-20 column chromatography was performed by adding a gradient to the water, and the second spot was obtained by prep-TLC (chloroform: methanol = 10: 1) to separate and purify 32 mg of chryseriol of Chemical Formula 3 below. In addition, the remaining major spots were once again purified by adding 100% ethanol to 10% gradient, lowering the water and performing Sephadex LH-20 column chromatography to separate and purify luteolin 73 mg, a compound of Formula 4 below.

<Formula 3>

<Formula 4>

It was confirmed that chryseriol and luteolin were separated into a single compound through TLC patterns, and are shown in FIGS. 3A and 4A. 4b to 4d.

< Experimental Example  1> Effect of the compounds according to the invention on irritable bladder disease

1) chrysriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound

Bladder was extracted from normal SD rats to create 2x5 mm smooth muscle slices, which were placed in an organ bath (Organ bath model) to determine the degree of contraction / relaxation of smooth muscle to the treated material. Recorded as.

Phenylephrine with Criesriol-6-C-β-boibinopyranosyl-7-O-β-glucopyranoside compound at 0.005, 0.05 and 0.5 mg / ml, respectively, and without treatment Changes in bladder smooth muscle contraction and relaxation in samples treated with different concentrations (alpha-1 adrenergic agonist (PE)) and acetylcholine (muscarinic, nicotinic receptor agonist, ACh) before treatment with the compound Was measured and the values are shown in FIGS. 5A and 5B.

As can be seen in FIGS. 5A and 5B, the chryseriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound is concentration dependent on the contraction of the bladder by PE and ACh. Markedly inhibited, especially in response to PE.

2) chryseryol compounds

In the same manner as in 1), the glycerol compound was treated with 0.005, 0.05 and 0.5 mg / ml, respectively, before and after treatment with phenylephrine (alpha-1 adrenergic agonist, PE) and acetylcholine (muscarinic, nicotinic receptor agonist, ACh). The change in the contraction and relaxation response of the bladder smooth muscle with respect to the concentration change of) is measured and the values are shown in FIGS. 6A and 6B.

As can be seen in FIGS. 6A and 6B, the chryseryol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside compound is concentration dependent on the contraction of the bladder by PE and ACh. Markedly inhibited, especially in response to PE.

3) Luteolin Compound

The luteolin compound was treated with 10 ^-6 to 10 ^-4 M in the same manner as in 1), before and after treatment with phenylephrine (alpha-1 adrenergic agonist, PE) and acetylcholine (muscarinic, nicotinic receptor agonist, ACh). The change in the contraction and relaxation response of the bladder smooth muscle with respect to the concentration change of was measured and shown in Figures 7a and 7b.

As can be seen in Figures 7a and 7b, the contraction response to PE and Ach was increased at low concentrations (10 ^-6 M ~ 10 ^-5 M) but inhibited at high concentrations (10 ^-4 M).

Thus, the chryseriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside, chryseriol and luteolin compounds according to the present invention are used for the bladder with phenylephrine and acetylcholine. It can be usefully used as a dietary supplement for improving the symptoms of patients with a tendency to overactive bladder and to treat urinary system diseases by inhibiting contraction.

< Experimental Example  2> Effect on the Prostate of the Compounds According to the Invention

Prostate tissue and prostate urethra from NewZealand white rabbits were extracted to make a 2x5 mm smooth muscle section, which is located in an organ bath (Organ bath model: quantifying the degree of contraction / relaxation of smooth muscle to the treated material). Chryseriol-6-C-β-boibinopyranosyl-7-O-β-glucopyranoside, chryserinol and lutein for each concentration of phenylephrine (alpha-1 adrenergic agonist, PE) The raised compounds were treated with 10 ⁻⁶ , 10 ^−5, and 10 ⁻⁴ M, respectively, to compare the reactions before and after the treatment and the values are shown in FIGS. 8 to 10, respectively.

As can be seen in Figures 8 to 10, the chryseriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside, chryserinol and luteolin compounds according to the present invention are Significantly suppresses the contraction caused by PE, so it has the effect of relaxing the urethra like alpha blocker commonly used in patients with enlarged prostate. .

< Formulation example  1> Preparation of Tablet

Chrysriol-6-C-β-vobinopyranosyl-7-O-β-glucopyranoside 60 mg

Calcium Silicate 20 mg

Lactose 130 mg

100 mg of microcrystalline cellulose

Magnesium stearate appropriate amount

Talc proper amount

Coating amount

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

< Formulation example  2> Preparation of capsule

Chryseriol 60 mg

Lactose 160 mg

Starch

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

< Formulation example  3> Manufacturing of beverage

0.26% by weight of honey, 0.0002% by weight of thioctoamide, 0.0004% by weight of nicotinic acid, 0.0001% by weight of sodium riboflavinate, 0.0001% by weight of pyridoxine hydrochloride, 0.001% by weight of inositol, 0.002% by weight of orthoic acid and 89.7362% by weight of water and luteolin A beverage was prepared by the conventional method by combining 10% by weight.

Claims (1)

An agent for treating prostatic hyperplasia or overactive bladder comprising the luteolin represented by the following formula (4) as an active ingredient.
<Formula 4>

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