US20100291154A1 - Composition for inhibition of transplant rejection containing the cordyceps mycellia extract as an active ingredient - Google Patents

Composition for inhibition of transplant rejection containing the cordyceps mycellia extract as an active ingredient Download PDF

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Publication number
US20100291154A1
US20100291154A1 US12/740,139 US74013910A US2010291154A1 US 20100291154 A1 US20100291154 A1 US 20100291154A1 US 74013910 A US74013910 A US 74013910A US 2010291154 A1 US2010291154 A1 US 2010291154A1
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cordyceps
composition
extract
mycellia
present
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US12/740,139
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English (en)
Inventor
Man Woo Han
Jae Kuk Yoo
Chang-Uk Hur
Hwan-Chul Kim
Jin Pyo Kim
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Hankook Pharm Co Inc
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Hankook Pharm Co Inc
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Assigned to HANKOOK PHARM. CO., INC. reassignment HANKOOK PHARM. CO., INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: HAN, MAN WOO, HUR, CHANG-UK, KIM, HWAN-CHUL, KIM, JIN PYO, YOO, JAE KUK
Publication of US20100291154A1 publication Critical patent/US20100291154A1/en
Priority to US13/161,958 priority Critical patent/US8206760B2/en
Abandoned 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
    • A61K36/062—Ascomycota
    • A61K36/066—Clavicipitaceae
    • A61K36/068—Cordyceps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents

Definitions

  • the present invention relates to a composition for inhibition of transplant rejection, comprising a cordyceps mycellia extract as an active ingredient.
  • Transplant rejection occurs when the immune system of the recipient of a transplant attacks a transplanted organ or tissue.
  • effective suppression of the immune response is known as a main factor determining the success of transplantation.
  • the development of immunosuppressive medications has brought about exceptional advances in the transplantation of organs and tissues and the treatment of autoimmune diseases and has made a great contribution to the study of the in vivo mechanism of immune responses to the transplanted organ or tissue.
  • immunosuppressive drugs were developed to inhibit or attenuate transplant rejection.
  • An example is cyclosporine A (U.S. Pat. No. 4,117,118) produced from Tolypocladium inflatum , a soil fungus.
  • These immunosuppressive drugs not only help realize clinically successful organ transplantation, but also suggest the therapeutic use thereof in treating autoimmune diseases. Even though they are required to act selectively and specifically for T-cells only, conventional immunosuppressive drugs have an influence on a wide range of cellular functions, including general signal pathways, causing side effects on other organs, which are healthy (see. S.-H. Lee et al., Korean J. Immunology, 19:375 ⁇ 389 (1997)).
  • cyclosporine A is known to show side effects of chronic liver diseases and hypertension after heart transplantation (see: J. E. F. Reynolds, et al., Martindale The Extra Pharmacopoeia, 31 st ed., pp. 557-562, Royal Pharmaceutical Society, London, 1996).
  • FK-506 has recently been discovered to be an immunosuppressant, and has been commercialized.
  • side effects of this drug have also been found (Clin. Transplantation, 11: 237 ⁇ 242 (1997)).
  • Vegetable worms are a kind of medicinal fungus produced as a result of the parasitism of vegetable worms on insects. In high temperature and moisture conditions, the vegetable worms, which are actually fungus, infect living insects, proliferate therein to kill the host insects, and form fruiting bodies on the surface of the host insects.
  • the term “vegetable worm”, is intended to primarily refer to Cordyceps sinensis , a parasite on larvae of the Hepialidae family, but at present generally refers to all fungi attacking arthropods, such as spiders.
  • Cordyceps sinensis breaks down into 10.8% water, 8.4% lipids, 25 ⁇ 32% crude proteins, 23.9% carbohydrates, and 18.5% crude fibers.
  • this vegetable worm contains 17 different amino acids, including 8 essential amino acids. Also, it contains a trace amount of cordycepin, 7.6% of D-mannitol and 11.2% of polysaccharides, all known as medicinally active materials.
  • Cordycepin a derivative of the nucleoside adenosine, is an isomer of quinic acid, known to show anti-cancer activity.
  • an immunosuppressive composition comprising a mycelial extract from a vegetable worm as an active ingredient is provided for the inhibition of transplant injection.
  • composition comprising a cordyceps mycellia extract as an active ingredient is provided for the prevention and treatment of skin diseases.
  • the cordyceps mycellia extract was found to significantly suppress the production of antibodies to transplants without side effects, such as weight change. Based on a natural material, the composition is non-toxic and harmless to the human body, and thus can be used as an immunosuppressant for organ transplantation. Also, it arrests oozing from sores and is applicable to the prevention and treatment of skin diseases, including atopy, allergic reactions, decubitus ulcers, pemphigus and smallpox.
  • FIG. 1 is a graph showing the levels of antibodies produced against splenocytes transplanted into mice administered with the cordyceps mycellia extract of the present invention or with saline in accordance with an embodiment of the present invention ( ⁇ : control, ⁇ : experimental group).
  • FIG. 2 is a graph showing changes in the weight of the mice administered with the cordyceps mycellia extract of the present invention and with saline in accordance with an embodiment of the present invention ( ⁇ : control, ⁇ : experimental group).
  • FIG. 3 shows the dermal states of mice administered with the cordyceps mycellia extract of the present invention and with saline in photographs (panel a: experimental group, panel b: control).
  • FIG. 4 is a graph showing the levels of antibodies produced against splenocytes transplanted into mice administered with the cordyceps mycellia extract of the present invention or with other comparative chemicals in accordance with another embodiment of the present invention.
  • FIG. 5 is a graph showing changes in the weight of the mice administered with the cordyceps mycellia extract of the present invention and with other comparative chemicals in accordance with another embodiment of the present invention.
  • an immunosuppressive composition based on a cordyceps mycellia extract is provided for the inhibition of transplant rejection.
  • the cordyceps mycellia extract may be obtained from cultured mycelia, or may be commercially available.
  • vegetable worm powder which is sold as a health aid food, may be used in the present invention, whether it comes from fruit bodies, mycelia, or a combination thereof.
  • Examples of the vegetable worm useful in the present invention include Cordyceps militaris, Cordyceps sinensis , which is parasitic on larvae of the Hepialidae family, Hymenostilbe odonatae, Cordyceps nutans, Tilachlidiopsis nigra, Paecilomyces japonica, Cordyceps tricentri , and Cordyceps sphecocephala .
  • the cordyceps mycellia extract according to the present invention was tested for immunosuppressive effect on smallpox mouse models. After the transplantation of splenocytes thereinto, the animal models were administered with the cordyceps mycellia extract. ELISA analysis on the animal models for the quantification of antibodies to the splenocytes indicated that the cordyceps mycellia extract of the present invention significantly inhibits the production of the antibodies (see FIGS. 1 and 4 ). Therefore, the cordyceps mycellia extract according to the present invention can be used as an immunosuppressant for the prevention of transplant rejection.
  • cordyceps mycellia extract can be used as an immunosuppressant for the prevention of transplant rejection without the occurrence of side effects, such as changes in weight, upon organ transplantation.
  • the present invention provides a composition for the prevention and treatment of skin diseases, comprising a cordyceps mycellia extract as an active ingredient.
  • the cordyceps mycellia extract according to the present invention was tested for therapeutic activity for skin diseases on smallpox mouse models. After the administration of the cordyceps mycellia extract thereinto, the smallpox mouse models were observed to stop oozing from the smallpox sores ( FIG. 3 ). In contrast, the control, which was not administered with the cordyceps mycellia extract of the present invention, was observed to ooze from the smallpox sores and remain depilated at the sores.
  • the cordyceps mycellia extract according to the present invention can be used for the treatment of skin diseases, such as atopy, allergic reactions, decubitus ulcers, pemphigus, smallpox, etc.
  • composition When used as medications or health aid foods, the composition may further comprise one or more active ingredients having a function similar to that of the cordyceps mycellia extract.
  • the cordyceps mycellia extract in accordance with the present invention can be administered orally or non-orally, and may be provided in general medicinal forms.
  • the cordyceps mycellia extract of the present invention may be used in oral or non-oral forms. It is usually formulated in combination with a diluent or an excipient, such as a filler, a thickening agent, a binder, a wetting agent, a disintegrant, a surfactant, etc.
  • Solid agents intended for oral administration of the extract of the present invention may be in the form of tablets, pills, powders, granules, capsules, and the like.
  • the cordyceps mycellia extract of the present invention is formulated in combination with at least one excipient, such as dextrin, starch, calcium carbonate, sucrose, lactose, or gelatine.
  • a lubricant such as magnesium stearate, talc, or the like, may also be added.
  • Liquid agents intended for oral administration include suspensions, internal use solutions, emulsion, syrups, and the like.
  • various excipients such as wetting agents, sweetening agents, aromatics, preservatives, and the like, may be contained in the liquid agents for the oral administration of the extract of the present invention.
  • non-oral dosage forms of the extract of the present invention include sterile aqueous solutions, non-aqueous solutions, suspensions and emulsions, freeze-dried agents, and suppositories.
  • Non-aqueous solutions and suspensions made from propylene glycol, polyethylene glycol, vegetable oils, such as olive oil, and injectable esters such as ethyl oleate may be used.
  • the basic materials of suppositories include Witepsol, macrogol, Tween 61, cacao butter, laurin, glycerol, and gelatin.
  • the effective dosage of the cordyceps mycellia extract in accordance with the present invention depends on various factors, including the patient's weight, age, gender, state of health, diet, the time of administration, route of administration, excretion rate, etc.
  • the cordyceps mycellia extract in accordance with the present invention may be administered at a dose ranging from 550 to 2,200 mg/day.
  • the cordyceps mycellia extract of the present invention may be used alone or in combination with other therapies, including surgery, radiotherapy, hormonal therapy, chemical therapy and/or biological reaction regulators.
  • mice From two days before the transplantation of splenocytes (Dsg3 ⁇ / ⁇ ), five smallpox mice were administered orally with a cordyceps mycellia extract powder (Cordyma®; Han Kook Sin Yak) at a dose of 10 mg/kg/day. For 35 days (5 weeks) after the splenocyte transplantation, the cordyceps mycellia extract was orally administered at a dose of 10 mg/kg/day. For a control, physiological saline was used instead of the extract.
  • a cordyceps mycellia extract powder Cordyma®; Han Kook Sin Yak
  • mice Blood samples were taken from the mice 7 days (1 week), 14 days (2 weeks), 21 days (3 weeks), 28 days (4 weeks) and 35 days (5 weeks) after the transplantation, and were analyzed using ELISA to determine the level of antibodies to the splenocytes. Throughout the experiment, the weights and skin states of the mice were monitored every day.
  • FIG. 1 the levels of antibodies to the splenocytes are plotted against time for the experimental group and the control group.
  • FIG. 2 shows changes in weight for the experimental group and the control group.
  • FIG. 3 the experimental group and the control group are compared with each other with respect to skin state.
  • the level of antibodies to the transplanted splenocytes in the mice administered with the cordyceps mycellia extract of the present invention was almost zero. From this, it is apparent that the cordyceps mycellia extract according to the present invention effectively inhibits the production of antibodies to transplants.
  • the weight of the control administered with physiological saline slightly increased immediately after the transplantation, but sharply decreased from 7 days after the transplantation, as transplant rejection occurred.
  • almost no change was found in the weight of the mice administered with the cordyceps mycellia extract of the present invention.
  • the composition comprising the cordyceps mycellia extract according to the present invention can therefore be used as an immunosuppressive medication applicable for organ or tissue transplantation.
  • cordyceps mycellia extract according to the present invention can be applied to the treatment of skin diseases including atopy, allergic reactions, decubitus ulcers, pemphigus, smallpox, etc.
  • mice Two S129 Dsg3 ⁇ / ⁇ mice and 24 S129 Rag2 ⁇ / ⁇ mice were prepared as transplantation donors and recipients, respectively.
  • the recipient mice were divided into four groups of six: control (CMC administered), comparative group (cyclophosphamide (CPA) administered), experimental group 1 and experimental group 2 ( cordyceps mycellia extract (Cordyma®; Han Kook Sin Yak) administered).
  • the transplantation donor was subjected to an immune reaction by administering a Dsg3-His protein thereto, as described below.
  • 10 ⁇ g of mouse Dsg3-His protein was emulsified with the same amount of complete Freund's adjuvant (CFA) and subcutaneously injected into the mice (1 st ).
  • CFA complete Freund's adjuvant
  • splenocytes From one day before the transplantation of splenocytes (Dsg3 ⁇ / ⁇ ), oral administration was conducted with 1 ml of the cordyceps mycellia extract (Cordyma®; Han Kook Sin Yak) for each experimental group, 1 ml of CMC for the control group and 1 ml of cyclophosphamide for the comparative group. Thereafter, splenocytes were transplanted at a density of 1.5 ⁇ 10 6 cells/500 ⁇ l into each mouse, followed by oral administration of 1 ml of the test material to the mice on day zero, 1, 4, 7, 11, 14, 18, 21, 25, and 28 after the transplantation.
  • mice were taken from the mice on day zero, 1, 4, 7, 11, 14, 18, 21, 25 and 28 after the transplantation, and were analyzed to determine the level of antibodies to the splenocytes using ELISA. Throughout the experiment, the weights and skin states of the mice were monitored every day.
  • the cordyceps mycellia extract (Cordyma®; Han Kook Sin Yak) for the experimental group 1 was prepared by placing 500 mg of a powder of Isaria japonica in a 15 ml tube, adding a 0.5% CM-Na solution (hereinafter referred to as “CMC”) to the tube to form a total volume of 5 ml, and sonicating it for 30 min in a bath to form a suspension (dosage: 2000 mg/kg/administration).
  • CMC CM-Na solution
  • cordyceps mycellia extract (Cordyma®; Han Kook Sin Yak) was prepared by mixing 0.5 ml of the cordyceps mycellia extract (Cordyma®; Han Kook Sin Yak) for the experimental group 1 with 4.5 ml of CMC and treating the mixture for 30 min in a sonication bath to give a suspension (dosage: 200 mg/kg/administration).
  • the cyclophosphamide administered to the comparative group was a suspension prepared by pulverizing 50 mg of the Endoxan P tablet, commercially available from Shionogi Pharmaceutical Corporation, mixing the powder with 16.7 ml of CMC, and sonicating the mixture in a bath for 30 min (dosage: 60 mg/kg/administration).
  • FIG. 4 the levels of the antibodies to the splenocytes were plotted against time for the experimental groups, the control group and the comparative group.
  • FIG. 5 shows changes in weight for the experimental groups, the control group and the comparative group.
  • the production level of antibodies to the transplanted splenocytes was observed to be lower in the mice administered with the cordyceps mycellia extract of the present invention than in the mice administered with CMC only (control).
  • ELISA values a Dunnett's multiple comparison test was performed on the basis of a PBS group. On Day 11 after the transplantation, there was significance for the experimental group 1 (p ⁇ 0.05), but not significance for the experimental group 2. On Day 14, there was significance for both the experimental group 1 (p ⁇ 0.01) and the experimental group 2 (p ⁇ 0.05). However, no significance was observed for either of them from Day 18.
  • the significant difference on Days 11 and 14 between the experimental group administered with the cordyceps mycellia extract at a dose of 2000 mg/kg and the control group indicates that the cordyceps mycellia extract of the present invention significantly inhibits the production of antibodies responsible for transplant rejection.
  • the cordyceps mycellia extract according to the present invention can be applied to the treatment of skin diseases including atopy, allergic reactions, decubitus ulcers, pemphigus, smallpox, etc.
  • composition of the present invention can be prepared as described below.
  • the cordyceps mycellia extract was dissolved in a suitable volume of an NaCl BP injection, and the solution was adjusted to a pH of 3.5 with diluted HCl BP and to a desired volume with NaCl BP injection, followed by sufficient mixing.
  • the solution was loaded into transparent ml type I ampules, which were hermetically sealed by melting, followed by autoclaving at 120° C. for 15 min to prepare injections.

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US12/740,139 2007-12-07 2007-12-07 Composition for inhibition of transplant rejection containing the cordyceps mycellia extract as an active ingredient Abandoned US20100291154A1 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9775868B2 (en) 2012-09-13 2017-10-03 Jiangzhong Pharmaceutical Co., Ltd. Traditional Chinese medicine combination for regulating immune function and preparation method therefor

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Publication number Priority date Publication date Assignee Title
CN102641339B (zh) * 2012-04-25 2014-03-05 吉林大学珠海学院 一种提高机体免疫力、抗疲劳度的中药复方制剂

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US4117118A (en) * 1976-04-09 1978-09-26 Sandoz Ltd. Organic compounds
US5948404A (en) * 1994-06-30 1999-09-07 Meiji Milk Products Company Limited Healthful composition obtained from the hot water extract of Coratceps sinensis mycelia

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JPH0940690A (ja) * 1995-05-23 1997-02-10 Yutaka Sashita ステロイド配糖体及びこれを有効成分とする医薬
JP3796780B2 (ja) * 1995-10-26 2006-07-12 味の素株式会社 新規免疫抑制剤
JP3621404B1 (ja) * 2003-09-25 2005-02-16 株式会社バイオソリューションズ 免疫力強化活性剤
KR100544945B1 (ko) * 2003-11-27 2006-01-24 박태선 밀리타리스동충하초 추출물을 유효성분으로 포함하는면역증진용 약학조성물
KR20060092373A (ko) * 2005-02-17 2006-08-23 (주)네오팜 소아 아토피 피부염의 예방 및 개선용 생약 조성물
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Publication number Priority date Publication date Assignee Title
US4117118A (en) * 1976-04-09 1978-09-26 Sandoz Ltd. Organic compounds
US5948404A (en) * 1994-06-30 1999-09-07 Meiji Milk Products Company Limited Healthful composition obtained from the hot water extract of Coratceps sinensis mycelia

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9775868B2 (en) 2012-09-13 2017-10-03 Jiangzhong Pharmaceutical Co., Ltd. Traditional Chinese medicine combination for regulating immune function and preparation method therefor

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US20110243970A1 (en) 2011-10-06
WO2009072688A1 (en) 2009-06-11
US8206760B2 (en) 2012-06-26
JP2011506308A (ja) 2011-03-03
KR20100112597A (ko) 2010-10-19
KR101266527B1 (ko) 2013-05-27
JP5872769B2 (ja) 2016-03-01

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