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WO2007078161A1 - Extrait de fomitopsis pinicola présentant une activité suppressive contre l'endommagement cellulaire pancréatique et utilisation de cet extrait - Google Patents

Extrait de fomitopsis pinicola présentant une activité suppressive contre l'endommagement cellulaire pancréatique et utilisation de cet extrait Download PDF

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Publication number
WO2007078161A1
WO2007078161A1 PCT/KR2007/000062 KR2007000062W WO2007078161A1 WO 2007078161 A1 WO2007078161 A1 WO 2007078161A1 KR 2007000062 W KR2007000062 W KR 2007000062W WO 2007078161 A1 WO2007078161 A1 WO 2007078161A1
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WO
WIPO (PCT)
Prior art keywords
extract
fomitopsis pinicola
pieces
composition according
fomitopsis
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2007/000062
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English (en)
Inventor
Seung-Hee Oh
Soon-Dong Kim
Sang-Il Lee
Hyun-Goo Lee
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Eugene Biofarm Coltd
Original Assignee
Eugene Biofarm Coltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
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Application filed by Eugene Biofarm Coltd filed Critical Eugene Biofarm Coltd
Publication of WO2007078161A1 publication Critical patent/WO2007078161A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus

Definitions

  • the present invention relates to a Fomitopsis pinicola extract and the use thereof.
  • the present invention relates to the fruit body extracts and cultured mycelial extract of Fomitopsis pinicola, which is effective in suppressing pancreatic injury.
  • pancreas running horizontally between the duodenum and the spleen, is an organ located in the digestive and endocrine system that serves to secrete several important hormones including insulin and glucagon into the blood and also serves to secrete pancreatic juice containing digestive enzymes such as amylase, lipase, trypsin, etc., and alkaline juices into the duodenum.
  • pancreatic injury results in the interruption of secretion of the digestive enzymes and the hormones responsible for the regulation of blood sugar level.
  • beta cells of the pancreatic islets of Langerhans are injured, diabetes mellitus and complications are likely to occur.
  • pancreas There are many factors that harm the pancreas, such as infection, autoimmune response, etc.
  • Various efforts have been made to treat injured pancreas, including the differentiation of stem cells into beta cells and the transplantation of pancreatic islets, but these are difficult to apply in practice.
  • Pancreatic islet transplantation encounters difficulties related to the undersupply of islets, side effects caused by the long-term administration of immunosuppressive agents, and destruction due to autoimmune response.
  • immunosuppressive agents agents that cause the long-term administration of immunosuppressive agents, and destruction due to autoimmune response.
  • differentiation and proliferation using embryonic stem cells ethical problems arise, and there is the possibility of the transplant, even if successfully achieved, differentiating into undesired cells and further into cancerous cells in other tissues or organs.
  • compositions effective in suppressing pancreatic injury induced by diabetes mellitus, comprising a fruit body extract or cultured mycelial extract of Fomitopsis pinicola.
  • the fruit body extract is a hot- water extract or an alkali extract.
  • the hot- water extract may be prepared by cutting Fomitopsis pinicola fruit bodies into fine pieces, pulverizing the pieces, heating the pulverized pieces in water, concentrating the aqueous solution, precipitating the solution with ethanol, and freeze- drying the precipitate, in order.
  • the pulverized pieces of the fruit bodies are heated at 100°C for 24 hours and the aqueous solution is concentrated at 40°C to one tenth of the initial volume.
  • the alkali extract may be prepared by cutting Fomitopsis pinicola fruit bodies into fine pieces, pulverizing the pieces, swelling the pulverized pieces in IN KOH, homogenizing the pieces, filtrating the homogenate, neutralizing the filtrate, washing the neutralized filtrate with distilled water, and drying the washed filtrate at 60°C, in order.
  • the pulverized pieces of the fruit bodies are mixed with IN KOH in a ratio of 1:1 (w/v), left to swell for 1 hour, homogenized, filtered through a 100 mesh sieve, and neutralized with cone. HCl.
  • the cultured mycelial extract may be prepared by seed- and sub-culturing mycelia of Fomitopsis pinicola, inoculating the mycelia in a sterile and cold potato medium, neutralizing the culture, precipitating with ethanol, and dialyzing the precipitate against distilled water.
  • the mycelia are inoculated in the potato medium to an amount of 2%
  • FIG. 1 shows optical microphotographs, magnified 200 times, of a normal control group (NC) of mice raised for 4 weeks on different diets, a pancreatic injury group (DM), a pancreatic injury-induced, hot-water extract-administered group (DM-WE) which was administered with 1% of the hot- water extract after treatment with STZ, a pancreatic injury-induced, alkali extract-administered group (DM-AE) which was administered with 1% of the alkali extract after treatment with STZ, and a pancreatic injury-induced, cultured mycelial extract-administered group (DM-CM) which was administered with 1 % of the cultured mycelial extract after treatment with STZ.
  • NC normal control group
  • DM-WE pancreatic injury-induced, hot-water extract-administered group
  • DM-AE pancreatic injury-induced, alkali extract-administered group
  • DM-CM pancreatic injury-induced, cultured myceli
  • the present invention is directed to an extract from the fruit body or cultured mycelia of Fomitopsis pinicola, and a composition comprising the extract as an active ingredient for suppressing pancreatic injury.
  • the extract from the fruit body of Fomitopsis pinicola may be prepared with hot water or alkali.
  • a hot- water extract from the fruit body of Fomitopsis pinicola can be obtained as follows.
  • the fruit body is finely cut, pulverized, immersed in water, and heated, followed by concentration and precipitation with ethanol.
  • the precipitate is then freeze-dried.
  • Water is preferably used in an amount of about 30 times the weight of the pulverized fruit body, but is not limited to that amount.
  • Heating is preferably conducted at 100°C for about 24 hrs, but is not limited thereto.
  • concentration it is preferable that the supernatant be evaporated at 40°C until the final volume is reduced to one tenth of the initial volume.
  • An alkaline extract from the fruit body of Fomitopsis pinicola can be obtained as follows. The fruit body is finely cut, pulverized and left to swell for 1 hr in IN KOH, followed by homogenization using a homogenizer. The homogenate is filtered through a 100 mesh sieve, neutralized with cone. HCl, washed with distilled water, and then dried at 60°C. For the swelling, it is preferable that IN KOH be used in a ratio of approximately 1:1 (w/v). This alkaline extraction, called HAS (homogenization after alkali swelling) method, is exceptionally improved in yield over conventional ones.
  • HAS homogenization after alkali swelling
  • the extract from Fomitopsis pinicola according to the present invention was assayed for suppressive activity against diabetes mellitus-caused pancreatic injury as follows. First, the extract was orally administered to rats in which diabetes mellitus had been induced with STZ (streptozotocin). After being raised, each rat was sacrificed to excise the pancreas and adjacent tissues, which were then fixed with a 10% formalin solution, dehydrated with alcohol, embedded in paraffin, and stained with HE. Under an optical microscope, the samples were observed to determine the suppressive effect of the extracts on pancreatic injury.
  • STZ streptozotocin
  • the Fomitopsis pinicola extract is analyzed for ingredients and the molecular weights thereof.
  • the Fomitopsis pinicola extract is fractioned and purified through DEAE-cellulose ion exchange resin and a Sepharose CL-4B gel column, followed by methylation analysis using gas chromatography.
  • Pohang City, Korea were inoculated on a YM agar plate (yeast extract: 0.5%(w/v), peptone: 0.5%(w/v), malt extract: 0.2%(w/v), glucose: 1.0%(v/v), agar: 2.0%(w/v), pH 6.5) and sub-cultured at 30°C every 15 days.
  • the mycelia were seed cultured in YM broth using a rotary agitator (150rpm).
  • the mycelia were inoculated in a sterilized (120°C, 30 min), cold potato medium (water: 16L, potato powder: 300g, glucose: 150g, peptone: O.lg) to an amount of 2% (v/v), and cultured at 30°C for 10 days in a rotary agitator operating at 150rpm with sterile air (10cm /min) provided thereto. Then, neutralization with sodium hydrogen carbonate (NaHCO ) to a pH of 6.5 preceded precipitation with 10 volumes of ethanol.
  • NaHCO sodium hydrogen carbonate
  • SD rats each having a body weight of 200+5 g, were divided into 5 groups of 7, which were respectively set as a normal control group (NC), a diabetes mellitus control group (DM), in which diabetes mellitus was induced by STZ, a diabetes mellitus- induced, hot-water extract-administered group (DM-WE) which was administered with 1% of the hot- water extract after treatment with STZ, a diabetes mellitus-induced, alkali extract-administered group (DM-AE) which was administered with 1% of the alkali extract after treatment with STZ, and a diabetes mellitus-induced, cultured mycelial extract-administered group (DM-CM) which was administered with 1 % of the cultured mycelial extract after treatment with STZ.
  • NC normal control group
  • DM-WE diabetes mellitus control group
  • DM-WE diabetes mellitus- induced, hot-water extract-administered group
  • DM-AE
  • Each of the groups was reared for four weeks according to the dietary schedules given in Table 1, below.
  • STZ 55mg/kg was intramuscularly injected. Animals which had a blood sugar level of 300 mg/dL, measured 48 hrs after injection with STZ, were regarded as having had pancreatic injury induced therein. Blood sugar levels were measured using a bio-sensor and a kit.
  • AIN-vitamin mix (mg/kg): thiamin hydrochloride 600, riboflavin 600, pyridoxine hydrochloride 700, nicotinic acid 3,000, calcium D-pantothenate 1,600, folic acid 200, D-biotin 20, vitamin B 12 2.5, vitamin A 400,000 IU, vitamin D3 100,000 IU, vitamin E 7,500 IU and vitamin K 75 were mixed to form a total weight of 1,000 g and finely powdered.
  • ⁇ NC normal control
  • DM diabetes mellitus control
  • DM-WE 1% of Fomitopsis pinicola fruit body hot-water extract was administered after treatment with STZ
  • DM- AE 1% of Fomitopsis pinicola fruit body alkali extract was administered after treatment with STZ
  • DM-CM 1% of the cultured mycelial extract of Fomitopsis pinicola was administered after treatment with STZ
  • mice After being fed with the experimental diets for 4 weeks, the animals were starved for 16 hrs with only water given thereto. They were etherized and subjected to laparotomy along the ventral median line to excise the pancreas and adjacent tissues which were then fixed with a 10% formalin solution, dehydrated with alcohol, embedded in paraffin, stained with HE, and observed under an optical microscope.
  • pancreas was observed to have typical morphology in the NC group which was fed the basic diet, as shown in FIG. 1.
  • the functional arrangement of secretory cells around blood vessels was normal with no injury detected therein.
  • the DM group which was treated only with STZ, was observed to show a homogeneous nucleus density in marginal cells (red), but to have serious swelling in central cells (black). Further, some cells underwent necrosis (blue).
  • Fomitopsis pinicola extracts of the present invention were identified as suppressors of pancreatic injury in diabetes mellitus-induced rats, as evident in Example 2. In order to examine the principle of suppressing diabetes mellitus-induced pancreatic injury, the Fomitopsis pinicola extracts were qualitatively analyzed.
  • DEAE-Cellulose (Cl " ) ion exchange resin and Sepharose CL-4B gel according to a well-known method (see, Lee Shin young, Kang, Tae soo, Structure Analysis of Antitumoral Exo-polysaccharide (BWS) obtained from submerged cultivation of
  • dextran having MWs of 2,000,000, 500,000, and 300,000 (Sigma) was used. Absorbance at 280 nm was utilized to detect the protein composition of the fractions. Six-carbon sugars were analyzed according to the anthrone method (see, Spiro RG, Analysis of sugars found in glycoprotein in Method in Enzymology, Academic Press, New York 8: 4-10, (1966)).
  • Fomitopsis pinicola extract one main component of the Fomitopsis pinicola extract was identified as ⁇ -l,3-glucano- ⁇ -l,6-heterogalactomannan, in which ⁇ - 1,3-glucan is linked to ⁇ -l,6-heterogalactomannan.
  • This complex polysaccharide was found to be a proteoglycan, ranging in molecular weight from 300,000 to 500,000, as measured by gel filtration.

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  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Engineering & Computer Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
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  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
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  • Medicines Containing Plant Substances (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention concerne des extraits de Fomitopsis pinicola et leur utilisation. Des extraits de sporophore et des extraits mycéliens cultivés de Fomitopsis pinicola inhibent efficacement la lésion pancréatique induite par le diabète sucré, ce qui permet de trouver diverses applications dans le développement d'aliments fonctionnels destinés à prévenir la lésion pancréatique résultant d'une infection ou d'une réponse auto-immune.
PCT/KR2007/000062 2006-01-05 2007-01-04 Extrait de fomitopsis pinicola présentant une activité suppressive contre l'endommagement cellulaire pancréatique et utilisation de cet extrait Ceased WO2007078161A1 (fr)

Applications Claiming Priority (2)

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KR10-2006-0001431 2006-01-05
KR1020060001431 2006-01-05

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WO2007078161A1 true WO2007078161A1 (fr) 2007-07-12

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4830371A (fr) * 1971-08-20 1973-04-21
KR20050060726A (ko) * 2003-12-17 2005-06-22 김천환 소나무잔나비버섯 추출물이 포함된 혈당강하제 및 그제조방법
WO2005067955A1 (fr) * 2004-01-06 2005-07-28 Paul Stamets Activite antimicrobienne de champignons medicinaux

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS4830371A (fr) * 1971-08-20 1973-04-21
KR20050060726A (ko) * 2003-12-17 2005-06-22 김천환 소나무잔나비버섯 추출물이 포함된 혈당강하제 및 그제조방법
WO2005067955A1 (fr) * 2004-01-06 2005-07-28 Paul Stamets Activite antimicrobienne de champignons medicinaux

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE MEDLINE [online] USUI T. ET AL.: "Investigation of the heterogeneity of heterogalactan from the fruit bodies of Fomitopsis pinicola, by employing concanavalin A-Sepharose affinity chromatography", XP003015393, Database accession no. (NLM6894749) *
J. OF BIOCHEMISTRY, JAPAN, vol. 89, no. 4, April 1981 (1981-04-01), pages 1029 - 1037 *

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