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Extracts of Cyathus Striatus Mushrooms (19) TZZ __T (11) EP 2 576 761 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C12N 1/14 (2006.01) A61K 36/07 (2006.01) 08.02.2017 Bulletin 2017/06 (86) International application number: (21) Application number: 11738493.3 PCT/IL2011/000434 (22) Date of filing: 02.06.2011 (87) International publication number: WO 2011/151831 (08.12.2011 Gazette 2011/49) (54) EXTRACTS OF CYATHUS STRIATUS MUSHROOMS, PHARMACEUTICAL COMPOSITIONS COMPRISING THEM AND A NEW CYATHUS STRIATUS STRAIN EXTRAKTE AUS CYATHUS-STRIATUS-PILZEN, PHARMAZEUTISCHE ZUSAMMENSETZUNGEN DAMIT UND NEUER CYATHUS STRIATUS-STAMM EXTRAITS DE CHAMPIGNONS CYATHUS STRIATUS, COMPOSITIONS PHARMACEUTIQUES LES COMPRENANT ET NOUVELLE SOUCHE DE CYATHUS STRIATUS (84) Designated Contracting States: (74) Representative: Dennemeyer & Associates S.A. AL AT BE BG CH CY CZ DE DK EE ES FI FR GB Postfach 70 04 25 GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO 81304 München (DE) PL PT RO RS SE SI SK SM TR (56) References cited: (30) Priority: 03.06.2010 US 351078 P • ROUMYANA D PETROVA ET AL: "Fungal substances as modulators of NF-[kappa]B (43) Date of publication of application: activation pathway", MOLECULAR BIOLOGY 10.04.2013 Bulletin 2013/15 REPORTS ; AN INTERNATIONAL JOURNAL ON MOLECULAR AND CELLULAR BIOLOGY, (73) Proprietor: Carmel-Haifa University Economic KLUWER ACADEMIC PUBLISHERS, DO, vol. 34, Corporation Ltd no. 3, 9 November 2006 (2006-11-09), pages 31905 Haifa (IL) 145-154, XP019531902, ISSN: 1573-4978 cited in the application (72) Inventors: • KANG H S ET AL: "Cyathuscavins A, B, and C, • FARES, Fuad new free radical scavengers with DNA protection 25155 Hourfish (IL) activity from the Basidiomycete Cyathus • SHARVIT, Lital stercoreus", BIOORGANIC & MEDICINAL 26317 Qiryat Motzkin (IL) CHEMISTRY LETTERS, PERGAMON, ELSEVIER • WASSER, Solomon P. SCIENCE, GB, vol. 18, no. 14, 15 July 2008 36812 Nesher (IL) (2008-07-15) , pages 4047-4050, XP022852894, ISSN: 0960-894X, DOI: 10.1016/J.BMCL.2008.05.110 [retrieved on 2008-06-03] Note: Within nine months of the publication of the mention of the grant of the European patent in the European Patent Bulletin, any person may give notice to the European Patent Office of opposition to that patent, in accordance with the Implementing Regulations. Notice of opposition shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention). EP 2 576 761 B1 Printed by Jouve, 75001 PARIS (FR) 1 EP 2 576 761 B1 2 Description etes vesicolor, Inonotus obliquus, Hypsizigus mar- moreus, and Flammulina velutipes) (Ikekawa, 2001). TECHNICAL FIELD [0005] Many edible mushrooms were used in tradition- al folk medicine includingLentinus edodes (shiitake [0001] The present invention relates to a new and dis- 5 mushroom). Grifola frondosa (maitake), Hericium eri- tinct variety of higher Basidiomycetes mushroom Cyat- naceus, Flammulina velutipes, Pleurotus ostreatus, and hus striatus HAI-1302, deposited under The Budapest Tremellamesenterica which are also sources of relatively Treaty with the Centralbureau voor Schimmelcultures pure bioactive compounds for medical usage, while other (CBS)under Accession No. CBS 126585 (hereinafter Cy- non-edible species, such as Ganoderma lucidum, Schiz- athus striatus CBS 126585), to extracts thereof, to phar- 10 ophyllum commune, and Trametes versicolor, are used maceutical compositions comprising the extracts and to only for their medicinal properties. the extracts for use in treatment of pancreatic and other [0006] Fungal metabolites have been gaining scientific types of cancer. The present invention further relates to interest as they were found to possess medicinal prop- a pure submerged mycelial culture ofCyathus striatus erties. Many studies on medicinal mushrooms proved CBS 126585. 15 their potential not only as dietary supplements and im- munoenhancers (Wasser & Weis, 1999a, b) but also as BACKGROUND ART modulators of various cellular responses (Zaidman et al., 2005). [0002] Natural products have been the most significant [0007] Thediversification ofmedicinal higher fungirep- source of drugs and drug leads in history. Many conven- 20 resents great potential for new drugs. As known, cancer tional therapies have been developed from nature-de- chemotherapy has relied mostly on cytotoxic drugs, rived materials. Their dominant role in cancer chemo- which inhibit tumor cell proliferation and cause cell death. therapeutics is clear, with about 74% of anticancer com- Cytotoxic activities against various tumor cell lines of pounds being either natural products or natural product- higher fungi secondary metabolites were widely investi- derived. It is estimated that approximately 25% of the 25 gated in the past decades. drugsprescribed worldwide, at present, come from plants [0008] Low-molecular-weight (LMW) substances with and 60% of anti-tumor/anti-infectious drugs, already on fungal origin can penetrate the cell membrane and inter- the market or under clinical investigations, are of natural fere, in particular, with cellular pathways linked to proc- origin (Wang et al., 2007). esses such as inflammation, carcinogenesis, cell differ- [0003] Higher Basidiomycetes mushrooms (HBM) rep- 30 entiation and survival, metastasis, etc. On the basis of resent a major and still largely unexploited source of new these facts, medicinal mushrooms were established as pharmaceutical products. Of approximately 15,000 a novel and promising source for natural therapeutics known species, 2,000 are safe for human consumption, that can be successfully applied in the treatment of dif- and about 650 of them possess medicinal properties. Of ferent diseases, including cancer (Smith et al., 2002). about 650 mushroom species with known medicinal35 Extracts from many mushrooms, as well as specific ac- properties, only about 20 species are in use at the tive substances isolated from such extracts (e.g. Yassin present. Most traditional knowledge about medicinal et al., 2003), have been shown to exhibit anti-cancer ac- properties of HBM arrives from the Far East (China, Ja- tivity. pan, Korea, and Russian Siberia). Many pharmaceutical [0009] For example, Muller et al. (2006) found that substances with potent and unique properties have re- 40 methanol:water extracts from fruiting bodies of G. luci- cently been extracted from mushrooms. Unique antican- dum induce apoptosis for each of the 26 panel of human cer remedies were prepared from these extracts such as cancer cell lines examined, in a dose- dependent man- polysaccharides lentinan, krestin, and schizophyllan (Mi- ner. zuno, 1999). [0010] Wu et al. (2006) used ethyl acetate extracts [0004] Presentstudies suggestthat extracts and active 45 from the mycelium of Cordyceps sinensis to show the agents obtained from HBM act as immunomodulators or inhibitory effect on various cancer cell lines. They pro- as biological response modifiers (BRM). They help the jected the inhibitory effect to ergosterol and related com- body to strengthen itself and fight off illness by maintain- pounds present in the extract. ing physiological homeostasis and restoring the body’s [0011] Lu et al. (2004) used ethanol:water extracts of Host Defense Potentiators (HDP), which can have im- 50 the Coriolus vesicolor fruiting bodies to inhibit the prolif- mune system enhancement properties. This benefit is eration of Burkitt’s lymphoma B- cell line (HL- 60) and one of the reasons why they are currently used as ad- humanacute promyelocytic leukemia celllines (NB- 4) ,in juncts to cancer treatment in many countries. In Japan, a dose-dependent manner, with more than 90% suppres- Russia, China, and the USA several polysaccharide an- sion, and no significant suppression on the proliferation ticancer and immunomodulating agents have been de- 55 of normal liver cell lines (WRL). veloped from the fruiting body, mycelia, and culture me- [0012] Pancreatic ductal adenocarcinoma (PDAC) is dium of various medicinal mushrooms ( Lentinus edodes, one of the deadliest of all solid malignancies. It is the Ganoderma lucidum, Schizophyllum commune, Tram- thirteenth most common cancer worldwide and one of 2 3 EP 2 576 761 B1 4 the leading causes of cancer death with 232,000 new Consequently, the biomass and extracts of this novel cases each year and 213,000 deaths. In the United mushroom is efficacious in treating cancer, in particular States and Israel, pancreatic cancer is the fourth leading pancreatic cancer, and can be used in pharmaceutical cause of cancer related mortalities. In the year 2010 an products. estimated 43,140 patients were diagnosed with pancre- 5 [0017] The present invention relates therefore, in one atic cancer; 36,800 patients died of their disease in the aspect, to the new and distinct variety of higher Basidi- US alone. omycetes mushroom Cyathus striatus HAI-1302, depos- [0013] PDAC is a disease with very poor prognosis ited under The Budapest Treaty with the Centralbureau mainly because of insensitivity to most standard thera- voor Schimmelcultures (CBS) under Accession No. CBS pies including chemotherapy, radiotherapy, and immu- 10 126585 (hereinafter Cyathus striatus CBS 126585), an notherapy. Therefore, surgical resection offers, at the extract obtained from Cyathus striatus CBS 126585, and moment, the only potential prospect for a cure. a pharmaceutical composition comprising a pharmaceu- [0014] For all stages combined, the 1- and 5-year rel- tically acceptable carrier and said extract. In certain em- ative survival
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