KR101880934B1 - Pharmaceutical composition containing platycodin D extracted from Platycodon grandiflorum and a method of producting thereof - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing a large amount of platycodin D extracted from a balloon flower, which generates a pharmaceutical composition containing a large amount of platycodin D extracted from a skin layer of balloon flower roots to prepare a medicine having excellent effects for anti-inflammatory, anticancer, and pain relief, and to a method for preparing the same. According to the present invention, an extract containing a large amount of balloon flower saponin is extracted from balloon flower roots in a large amount, and an ultrafiltration membrane is used from the extract, and thus, particularly, balloon flower saponin containing a large amount of platycodin D can be concentrated to be produced.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for preparing a pharmaceutical composition containing a blooming saponin concentrate extracted from a bellflower,

The present invention relates to a pharmaceutical composition containing a large amount of platycodine D extracted from a bellflower, and a method of preparing the same. More specifically, the present invention relates to a pharmaceutical composition containing a large amount of platidone D And a method for producing the pharmaceutical composition, which comprises a large amount of platycodine D extracted from bellflower, which is capable of producing a drug having excellent anti-inflammatory, anti-cancer, and pain relieving effects.

Platycodon grandiforum A. DC) is a perennial herbaceous plant belonging to Campanualaceae that grows or grows in East Asia. Its roots are mainly used as traditional herb medicine and food.

The main ingredient of the bellflower is carbohydrate (sugar), which accounts for more than 90%, and other proteins, fats, and ashes. Carbohydrates are mostly disaccharides or trisaccharides such as glucose, fructose, sugar, and cestose, and polysaccharides such as inulin, platycodinin (polysaccharide of about 10 molecules of fructose) have been reported. In addition, about 2% of triterpenoid saponin (24 kinds including platycodin A, C, D, D2, polygalacin D and D2 are reported) , And these saponin components are attracting attention as effective ingredients for various pharmacological effects of bellflower.

In addition, the bellflower contains steroids such as α-spinasterol, Δ7-stigmasterol, α-spinasteryl-β-D-glucoside, 0.0 > 0.03% < / RTI > of a steroid compound.

Platycodon has been used mainly as a remedy for gadam, deep-sea, cough, and bronchitis, and has anti-inflammatory effects [Biol. Pharm, Bull., 11, p.2114, 2008; Eur. J. Pharmacol., 537, p. 1, 2006; Life sci., 76, p. 2315, 2005], anti-cancer effect [Int. J. Cancer, 261, p. 2667, 2008; Cancer Lett., 261, p. 88, 2008], pain relief effects [see: Am. Chin. Med., 32, p. 257, 2004; Planta Med., 68, p. 794, 2002], inhibitory effect on digestive enzyme secretion (Arch.Pharm. Res., 27, p. 1048, 2004], cholesterol metabolism improvement effect [Eur. J. Pharmacol., 537, p. 1, 2006; J. Food Sci., 73, p. 195, 2008], protection and improvement of liver function [Lee, K.J. et. al., Toxicol. Lett. 147, 271-282, 2004; Food and Chemical Toxicology, 47, 2749-2754, 2009; Ann Nutri Metab 58, 224-231, 2011], Immune function modulating activity [Ahn, K.S. et al., Life Sci. 76, 2315-2328, 2005].

In addition, neuroprotective effect [Yoo Ki-Yeon et al., Neurosci. lett. 444 (1), 97-101 (2008)], anti-obesity effect [Zhao, H. L. et al., J. Food. Sci. 73 (8), H195-H200, (2008)]. Particularly, platycodin D is a saponin having a main pharmacological component of Gil-gye (a medicinal substance made from a rootstock of rosewood), and has anti-apoptosis (Korean Patent Registration No. 10-0564927), anti-obesity, Korean Patent Registration No. 10-0750333) and anti-inflammation (Korean Patent Registration No. 10-0696647) are known, and a single component having a high pharmacological value (Hahn J., Nutr., 130, p .2760, 2000).

In order to efficiently isolate compounds exhibiting pharmacological activity from such natural products, many researchers have developed and reported various methods for their extraction, separation and purification processes.

In general, the separation of natural components is carried out by using column chromatography using various types of solid phase and liquid phase. However, this method has limitations on the amount that can be separated at one time, and because of the cost of the equipment and the periodic replacement of the solid stationary phase, it is more expensive than the amount of separation and it is difficult to industrialize mass separation and production methods.

[Reference 1] Yoo Ki-Yeon. et al., Neurosci. lett. 444 ([0011] 1), 97-101, (2008)

[Reference 2] Zhao, H. L. et al., J. Food Sci. 73 (8), H195-H200, (2008)

[Reference 3] Lee, K. J. et al., Toxicol. Lett. 147, 271-282 (2004)

[Reference 4] Ahn, K. S. et al., Life Sci. 76, 2315-2328, (2005)

[Reference 5] Lee, K. J. et al., Food Chem. Toxicol. 47, 2749-2754, (2009)

[Reference 6] Zhang, L. et al., Molecules 12 (4), 832-841, (2007)

KR 10-2010-0069202 A KR 10-2010-0086923 A KR 10-2011-0061976 A

In order to solve the above-mentioned problems, the present invention is to maximize the extracting amount of borax saponin in a proper amount of water and an organic solvent and to extract borax saponin containing a large amount of platycodine D from these extracts using an ultrafiltration membrane And to provide a pharmaceutical composition containing a large amount of platycodine D extracted from a bellflower which can be concentrated in a large amount, and a method for producing the same.

In addition, platycodin D extracted from bellflower, which is obtained from bellflower in a simple and economical manner in a short period of time in a short period of time by obtaining a large amount of platycodin D containing platycodine D in a large amount to maximize the pharmacological effect, And a method for producing the same.

Disclosed is a method for preparing a pharmaceutical composition containing platycodin D, comprising the steps of: preparing dried bellflower in powder form; A second step of adding a solvent to the bellflower powder to produce a bellflower extract; A third step of cooling the bellflower extract; A fourth step of passing the bellflower extract through a microfilter to remove insoluble matter; A fifth step of passing the bellflower extract from which the insoluble matter is removed through an ultrafiltration membrane to remove low molecular weight sugar and water to produce a bellows saponin concentrate; A sixth step of concentrating the blooming saponin concentrate by heating under reduced pressure; And a seventh step of pulverizing the dried blooming saponin concentrate to obtain a powder, and preparing a pharmaceutical composition containing the powder.

The bellflower is characterized by separating the whole bellflower roots or the cork layer, and then preparing it by slicing.

The solvent to be added in the second step is one of water, an organic solvent, a mixture of water and the organic solvent, and the organic solvent is a lower alcohol having 1 to 4 carbon atoms including methanol, ethanol and butanol .

The ultrafiltration membrane used in the fifth step has a molecular weight limit of 1,000 or more.

The bellflower saponin concentrate may contain at least one selected from the group consisting of Deapioplatycoside E, Platycoside E, Platycodin D3, Polygalacin D2, Polygalacin D, D), Platyconic acid A, Deapioplatycodin D2, Platycodin D2, Deapioplatycodin D, Platycodin D, D), 2 "-O-acetyl-deapiopolygalacin D2, 2" -O-acetyl-polygalacin D2, And platyconic acid A methyl ester, Platycodigenin, Polygalacic acid, Platycogenic acid A lactone, Platycogenin A lactone, Platycogenic acid A lactone 3-O-glucopyranoside, Platycodigenin 3-O-Glucopyranoside 28-methyl ester ( Platycodigenin 3-O-glucopyranoside 28-methyl ester) and platycodigenin 3-O-laminaribioside 28-methyl ester. .

And is a pharmaceutical composition containing a large amount of platycodine D extracted from the bellflower produced by such a manufacturing method.

It is possible to concentrate and produce blooming saponin containing a large amount of platycodine D, particularly by extracting a large amount of the extract containing a large amount of borosilicate saponin from the borax root according to the present invention and using the ultrafiltration membrane from the extract.

Figures 1A and 1B are chromatograms of extracts prepared by the method according to an embodiment of the present invention.
2 is a flowchart showing a process of a manufacturing method according to an embodiment of the present invention.

Hereinafter, a pharmaceutical composition containing a large amount of platycodine D extracted from platycodon and a method for producing the same will be described with reference to the drawings.

FIGS. 1A and 1B are chromatograms of an extract prepared by the method of the present invention, and FIG. 2 is a flowchart illustrating a process of a method according to an embodiment of the present invention.

A pharmaceutical composition containing a large amount of platycodine D extracted from a bellflower (hereinafter referred to as a " composition ") to be produced in the present invention has anti-inflammatory, anti-cancer effect, pain relief effect, digestive enzyme secretion, cholesterol It is a phylloxera extract with pharmacological effect such as metabolism improvement effect, liver function protection and improvement effect, immune function regulating activity, neuronal cell protection, anti-obesity, anti-cell death.

In order to produce a composition having pharmacological effects with such a purpose, the present invention uses a water, an organic solvent or a mixture thereof as a solvent to obtain a bellflower extract from the bellflower roots, and the bellflower extract thus obtained is concentrated using a ultrafiltration membrane Water is obtained, and a step of drying the concentrate of bellows saponin so as to be easily used as a health functional food or a pharmaceutical composition to prepare a powder. This process is used to provide a production method of extracting a large amount of borax saponin compound, concentrating and drying the borax saponin compound.

Hereinafter, a method for producing the composition of the present invention will be described with reference to FIG.

First, the dried bellflower is pulverized into a powder state. (102) Here, it is preferable that the bellflower powder is obtained by separating the whole bellflower roots or the cork layer, and then slicing and drying the bellflower. In the case of using the ballad in powder form, there is an advantage that the content eluted at the time of extraction is increased.

Then, a solvent (water, an organic solvent, or a mixture thereof) is added to the prepared bellflower powder (S104). As the organic solvent, a lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol, and butanol may be used. % To 100% concentration solution is used. The substance contained in the bellflower powder is extracted into the solvent. In the present invention, it is defined as a bellflower extract.

The solvent is added in an amount of 1 to 30 times the volume (w.v%), preferably 5 to 10 times the volume (w.v%) of the dried bellflower powder. The mixture of the bellflower powder and the solvent is subjected to extraction at a temperature of from 10 캜 to 90 캜, preferably at room temperature. The extraction is carried out once to 5 times by any one of the methods of cold extraction, hot water extraction, ultrasonic extraction, reflux extraction and heat extraction for 1 to 24 hours, preferably 6 to 24 hours. Preferably, it is extracted once to three times with room temperature extraction.

The platycodone extract obtained in the above process is cooled (S106)

More than 90% of the bellflower extracts made by this process are low molecular weight (less than 1 to 5) per 1,000 molecular weight. In addition, less than 2% of the solids of the bellflower extract is platycodon saponin, and platycodin D does not exceed 0.65% of the solids content of the bellflower extract. Therefore, it is necessary to remove the low molecular weight sugar which does not have such pharmacological effect and to concentrate the phloem saponin having pharmacological effect.

In order to remove small amounts of plant cell wall components or insoluble matters such as cellulose that may be present in the bellflower extract, a micro filter having a pore size of about 1 탆 is used for filtering (S 108)

Next, the bellflower extract from which the insoluble matter is removed is passed through an ultrafiltration membrane to concentrate while removing the low molecular weight and water (S110). Preferably, the ultrafiltration membrane having a molecular weight limit of 1,000 or more is permeated through the molecular weight property of the bellflower saponin Most of the unnecessary monosaccharides, disaccharides, trisaccharides and the like present in the bellflower extract are removed together with water or a solvent component. Since the ultrafiltration membrane has a molecular weight limit of 1,000 or more, the blooming saponin present in the bloom extract concentrate also has a molecular weight of 1,000 or more.

A state in which insoluble matter is removed while passing through a microfilter, and low molecular weight and water are removed while passing through an ultrafiltration membrane is defined as a bloom saponin concentrate.

The saponin-based compounds contained in the concentrate of the blooming saponin having been subjected to the above-mentioned processes are selected from the group consisting of Deapioplatycoside E, Platycoside E, Platycodin D3 and Polygalacin D2 Polygalacin D2, Polygalacin D, Platyconic acid A, Deapioplatycodin D2, Platycodin D2, and Diapioplatidone D Deapioplatycodin D, Platycodin D, 2 "-O-acetyl-deapiopolygalacin D2, 2" -O-acetyl-polygalacin D2 2 "-O-acetyl-polygalacin D2) and Platyconic acid A methyl ester, Platycodigenin, Polygalacic acid, Platycogenic acid A lactone, Platycogenic acid A-lactoprotein 3-O-glucopyranoside, Platycodega Platycodigenin 3-O-glucopyranoside 28-methyl ester and Platycodigenin 3-O-laminaribioside 28-methyl ester. methyl ester).

In the next step, such a concentrate is subjected to a concentration process again while heating it under reduced pressure. (S112) Preferably, the brix of the concentrate at 60 ° C to 80 ° C is preferably 10 to 30, It is better to avoid excessive heating in order to minimize the loss of bellflower saponin.

Finally, the blooming saponin concentrate is powdered to be easily usable as a health functional food or a pharmaceutical composition (S114)

The concentrated blooming saponin concentrate at an optimum concentration for drying can be easily removed by a method such as spray drying or freeze drying. And may be prepared as a powder and easily used as a raw material for a health functional food or a pharmaceutical composition.

Hereinafter, an embodiment in which a pharmaceutical composition is prepared by extracting a large amount of platidone D from borax saponin prepared through the above-described process will be described.

Reference Example 1  : HPLC (High Performance Liquid Chromatography) analysis

A mixture of water (70%) and acetonitrile (30%) was used for the HPLC to identify the components of the bell jar saponin, and a pump, automatic injector, vaporization light scattering detector (ELSD, shimazu, Was used as a mobile phase and a single bellflower saponin was separated using a reversed phase C18 column (Zorbax C18, 150 x 4.6 mm, 3.5 μm, 30 ° C) at a flow rate of 0.8 ml / min and a retention time of 13 min. . Platycodin D, which has a high pharmacological value, was used as an index material to measure the content of borage saponin.

Example 1  : Manufacture of Bellflower Extract

2,000 ml of distilled water was added to 100 g of bellflower powder and extracted once at 80 DEG C for 8 hours and then cooled to obtain a bellflower extract.

100 g of bellflower powder was mixed with an organic solvent (ethanol) and water to make 2,000 ml of each, and once extracted for 8 hours in a water bath, the mixture was cooled to obtain a bellflower extract.

In order to increase the content of the extract, the extract was used as the powder. The extract was used to filter the insoluble matter by primary filtration.

The solids content of each of the bellflower extracts was about 4 brix and was extracted up to about 80% of the solid content of the bellflower. Table 1 shows the content of platycodine D, an indicator component of bellflower saponin.

Example 2  : Extract of the raw material of the bellflower Platycodine  D content change

Based on the same results as in Example 1, the extraction amount according to the raw materials was checked to confirm the maximum extraction amount of the borax saponin according to the condition of the borax root, and the Borax extract was obtained as shown in Table 2.

The extract was obtained by extracting 10% of the organic solvent (ethanol) and 90% of water, which showed the best yield in Example 1, at room temperature for 8 hours. The results show that extraction at room temperature using a solvent containing 10% organic solvent is most effective by utilizing the cortex of Rhododendron root, but it is not necessarily limited to the ratio of these solvent components.

Example 3  : Ultrafiltration membrane  From the used bellflower extract, borage saponin Concentrate Manufacturing

The bellflower extracts obtained in Example 1 and Example 2 were extracted from bellflower to about 80% of the solids. However, in this extract, 90% or more of the bellflower extract had a molecular weight of 1,000 or less (per 1 to 5). The low molecular weight sugar was removed, and the bellflower extract was concentrated using a microfilter and an ultrafiltration membrane to remove insoluble matter and water.

The concentrated results are shown in Table 3 below.

As shown in Table 3, it was confirmed that platycodin D was concentrated more than twice. When chromatograms analyzed by HPLC (see FIG. 1A) were observed, other blooming saponins also showed the same pattern as platycodine D And the low molecular weight saccharides were removed with water.

Example 4  : Industrial Ultrafiltration membrane  Used bellflower saponin Concentrate  Produce

In order to confirm the practical industrial production possibility, 600 L of bellflower extract obtained by using the method of producing the maximum bellflower extract obtained in Example 1 and Example 2 was applied to an industrial equipment having a pre-filter having a pore size of 1 μm filtration system, MWCO 1,000) to a concentration of 6-fold. As a result of the concentration, the sugar content of the bellflower was enriched from 4brix to 9.6brix and platycodine D was concentrated from 250ppm to 2,603ppm. It was confirmed by HPLC chromatography (see FIG. 1B) that other bell jar saponins were also concentrated in the same manner, and the low-molecular sugar was confirmed to be removed with water relatively.

Example 5  : Bellflower saponin Concentrate  Spray drying

The bellflower saponin obtained in Example 4 was concentrated to 25 brix which is easy to spray dry at 60 ° C or lower in a heating and pressure-concentrator, and then spray-dried. The spray dried powder was analyzed by HPLC, and the concentration of platycodin D and the sugar content of the concentrate of the crude Drosophila saponin were found to be almost unchanged. Drosophila saponin exists as a triterpenoid component in which saccharides are bonded to each other. Since these saponins are relatively stable to heat, they can be easily powdered by freeze drying or spray drying. Repeated experiments using these industrial equipments resulted in the concentration of platycodine D in the final powder to 1 to 3% (w / w).

As can be seen from the above examples, phloem saponin having pharmacological effect from bellflower can be effectively extracted and powdered, and this powdered concentrate has a concentration (0.1 Weight% to 20% by weight), it can be variously formulated in the form of tablets, capsules, granules and the like. In addition, it has a sufficient value to be utilized as a raw material capable of exhibiting sufficient efficacy even with a small amount of intake, and it has become possible to continuously produce products continuously and repeatedly by the above method, thereby establishing a method of economically mass production.

While the present invention has been described with reference to exemplary embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, As will be understood by those skilled in the art. Therefore, it should be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than the foregoing description, It is intended that all changes and modifications derived from the equivalent concept be included within the scope of the present invention.

Claims (6)

A method for producing a pharmaceutical composition containing a bellows saponin concentrate,
A first step of drying and crushing a cortex layer formed from cork cores in powdered state;
A second step of adding a solvent mixed with 90% of water and 10% of ethanol to the bellflower powder to produce a bellflower extract;
A third step of cooling the bellflower extract;
A fourth step of passing the bellflower extract through a microfilter to remove insoluble matter;
A step 5 of passing the bellflower extract from which the insoluble matter has been removed through an ultrafiltration membrane having a molecular weight limit of 1,000 or more to remove low molecular weight sugar and water to produce a bellows saponin concentrate;
A sixth step of concentrating the bellflower saponin concentrate by heating under reduced pressure at a temperature of 60 ° C to 80 ° C;
7. A method for producing a pharmaceutical composition comprising a bellflower saponin concentrate extracted from a bellflower, comprising the steps of: pulverizing the dried bellflower saponin concentrate to form a powder, and then preparing a pharmaceutical composition containing the powder; . delete delete delete The method according to claim 1,
The bellflower saponin concentrate may contain at least one selected from the group consisting of Deapioplatycoside E, Platycoside E, Platycodin D3, Polygalacin D2, Polygalacin D, D), Platyconic acid A, Deapioplatycodin D2, Platycodin D2, Deapioplatycodin D, Platycodin D, D), 2 "-O-acetyl-deapiopolygalacin D2, 2" -O-acetyl-polygalacin D2, And platyconic acid A methyl ester, Platycodigenin, Polygalacic acid, Platycogenic acid A lactone, Platycogenin A lactone, Platycogenic acid A lactone 3-O-glucopyranoside, Platycodigenin 3-O-Glucopyranoside 28-methyl ester ( Platycodigenin 3-O-glucopyranoside 28-methyl ester) and platycodigenin 3-O-laminaribioside 28-methyl ester. ≪ / RTI > characterized in that it comprises a bloom saponin concentrate extracted from a bellflower. delete

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