(19) TZZ Z_ __T (11) EP 2 017 251 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: C07C 41/46 (2006.01) A61K 8/35 (2006.01) 19.07.2017 Bulletin 2017/29 A61K 9/48 (2006.01) A61K 9/50 (2006.01) A61K 31/122 (2006.01) A61P 3/02 (2006.01) (2006.01) (2006.01) (21) Application number: 07742687.2 C07C 41/26 C07C 41/34 C07C 43/23 (2006.01) (22) Date of filing: 27.04.2007 (86) International application number: PCT/JP2007/059252 (87) International publication number: WO 2007/126083 (08.11.2007 Gazette 2007/45) (54) METHOD FOR STABILIZATION OF REDUCED COENZYME Q10 VERFAHREN ZUR STABILISIERUNG VON REDUZIERTEM COENZYM Q10 PROCÉDÉ SERVANT À STABILISER UN COENZYME Q10 RÉDUIT (84) Designated Contracting States: (74) Representative: Hoffmann Eitle AT BE BG CH CY CZ DE DK EE ES FI FR GB GR Patent- und Rechtsanwälte PartmbB HU IE IS IT LI LT LU LV MC MT NL PL PT RO SE Arabellastraße 30 SI SK TR 81925 München (DE) (30) Priority: 28.04.2006 JP 2006126897 (56) References cited: EP-A1- 1 415 970 EP-A1- 1 452 174 (43) Date of publication of application: WO-A1-03/032967 21.01.2009 Bulletin 2009/04 • MATSURA T. ET AL.: ’Antioxidant role of cellular (73) Proprietor: KANEKA CORPORATION reduced coenzyme Q homologs and Osaka (JP) alpha-tocopherol in free radical-induced injury of hepatocytes isolated from rats fed diets with (72) Inventors: different vitamin E contents’ BIOCHIMICA ET • UEDA, Takahiro BIOPHYSICA ACTA vol. 1127, 1992, pages 277 - Kobe-shi, Hyogo 655-0872 (JP) 283, XP003018946 • KITAMURA, Shiro Akashi-shi, Hyogo 673-0882 (JP) Remarks: • KUBO, Hiroshi Thefile contains technical information submitted after Kobe-shi, Hyogo 655-0046 (JP) the application was filed and not included in this • HOSOE, Kazunori specification Takasago-shi, Hyogo 676-0025 (JP) 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 2 017 251 B1 Printed by Jouve, 75001 PARIS (FR) EP 2 017 251 B1 Description Technical Field 5 [0001] The present invention relates to a method of stabilizing reduced coenzyme 10Q, and a reduced coenzyme Q10-containing composition stable against oxidation. As compared to oxidized coenzyme Q 10, reduced coenzyme Q10 shows high oral absorbability, and is a compound useful as a superior food, food with nutrient function claims, food for specified health use, nutritional product, nutritional supplement, animal drug, drink, feed, pet food, cosmetic, pharma- ceutical product, therapeutic drug, prophylactic drug and the like. 10 Background Art [0002] It is known that reduced coenzyme Q10 can be obtained, for example, by a method comprising producing coenzyme Q10 by a conventionally known method such as synthesis, fermentation, extraction from a naturally occurring 15 substance and the like, and concentrating a reduced coenzyme Q 10 fraction in an eluate from chromatography and the like (patent reference 1). The patent reference 1 describes that, in this case, oxidized coenzyme Q10 contained in the above-mentioned reduced coenzyme Q10 may be reduced with a general reducing agent such as sodium borohydride, sodium dithionite (sodium hydrosulfite) and the like, and concentrated by chromatography, and that the reduced coen- zyme Q10 can also be obtained by a method comprising reacting existing highly pure coenzyme Q10 with the above- 20 mentioned reducing agent. [0003] In addition, production methods for conveniently obtaining reduced coenzyme Q10 are also disclosed (e.g., patent references 2 - 4). [0004] However, reduced coenzyme Q10 is easily oxidized by molecular oxygen into oxidized coenzyme Q10, and therefore, stabilization of reduced coenzyme Q 10 is an important issue when it is processed into a food, food with nutrient 25 function claims, food for specified health use, nutritional product, nutritional supplement, animal drug, drink, feed, pet food, cosmetic, pharmaceutical product, therapeutic drug, prophylactic drug and the like, or a material or composition therefor, or preserved after processing and the like. Complete removal or blocking of oxygen during the above-mentioned processing and preservation is extremely difficult, and remaining or admixed oxygen particularly during heating for processing and long-term preservation exerts a markedly adverse effect. The above-mentioned oxidation is directly 30 related to quality problems such as the by-product oxidized coenzyme Q10. [0005] As mentioned above, stabilization of reduced coenzyme Q10 (protection of oxidation) is a highly important object. However, since reduced coenzyme Q10 is not commercially available to date, the study of methods and compo- sitions for stable retention of reduced coenzyme Q10 has not been undertaken very much. [0006] As a conventionally-known method for stably retaining reduced coenzyme Q 10, a method including addition of 35 a reducing agent is known. However, some of the reducing agents used therefor are not suitable for food and pharma- ceutical agents. For example, patent reference 5 disclosing a composition concurrently containing a reducing agent and a production method thereof also discloses, (1) a composition comprising reduced coenzyme Q 10; a reducing agent in an amount effective for eliminating oxidation of reduced coenzyme Q 10 into oxidized coenzyme Q10; a surfactant, veg- etable oil or a mixture thereof in an amount effective for dissolving the above-mentioned reduced coenzyme Q10 and 40 the above-mentioned reducing agent; and a solvent as necessary, (2) a composition for oral administration wherein the above-mentioned composition is prepared into a gelatin capsule or a tablet, and (3) a method of preparing the above- mentioned composition containing reduced coenzyme Q10 in situ using oxidized coenzyme Q10 and a reducing agent. However, no detailed description relating to the quality, stabilizing effect and the like of the reduced coenzyme10 Q contained in the composition is provided, and the expected level of stabilization is not clear. 45 [0007] In addition, the above-mentioned composition and preparation method thereof are highly complicated and complex since plural roles are conferred to the composition (i.e., firstly, a role as a reaction site for reducing oxidized coenzyme Q10 to reduced coenzyme Q10, and secondly, a role of stably retaining reduced coenzyme Q10). Moreover, the above-mentioned composition and a preparation method thereof are not entirely safe because the reaction mixture is used as it is. In other words, ascorbic acids to be used as reducing agents are oxidized to produce a considerable 50 amount of dehydroascorbic acids, and the dehydroascorbic acids get mixed in with the above-mentioned composition, posing a problem. Dehydroascorbic acids and oxalic acid produced by decomposition from dehydroascorbic acids are highly noxious, unlike ascorbic acids. For example, an increased amount of lipid peroxide, a decreased amount of antioxidants in the liver and kidney and an increased amount of oxalic acid in the kidney have been reported, and side effects such as decreased resistance to oxidation stress, easy onset of ureteral lithiasis (non-patent reference 1) and 55 the like are feared. patent reference 1: JP-A-10-109933 patent reference 2: WO03/06408 2 EP 2 017 251 B1 patent reference 3: WO03/06409 patent reference 4: WO03/32967 patent reference 5: WO01/52822 non-patent reference 1: Nutrition Research Vol. 13, page 667-676, 1993 5 Disclosure of the Invention Problems to be Solved by the Invention 10 [0008] In view of the above, the present invention aims at providing a convenient and preferable method and a com- position for stably retaining reduced coenzyme Q 10 by protection against oxidation while maintaining high safety, during processing into a food, food with nutrient function claims, food for specified health use, nutritional product, nutritional supplement, animal drug, drink, feed, pet food, cosmetic, pharmaceutical product, therapeutic drug, prophylactic drug containing reduced coenzyme Q 10 and the like, or a material or composition therefore, and/or preservation after process- 15 ing and the like. Means of Solving the Problems [0009] The present inventors have conducted intensive studies in an attempt to solve the above-mentioned problems 20 and found that reduced coenzyme Q10 can be stabilized by the co-presence of reduced coenzyme Q9 and/or reduced coenzyme Q11, which are analogs of reduced coenzyme Q 10 and reduced coenzyme Q10. [0010] That is, they have found that reduced coenzyme Q 10 can be stably retained by protecting the reduced coenzyme Q10 from oxidation by the co-presence of reduced coenzyme Q 9 (not less than 1.5 wt% to not more than 99 wt% relative to reduced coenzyme Q10) and/or reduced coenzyme Q11, even when a reducing agent is not used as a necessary 25 component to be added, which resulted in the completion of the present invention. [0011] Accordingly, the present invention provides the following. [1] A method for stabilizing reduced coenzyme Q 10, comprising achieving the co-presence of the reduced coenzyme Q10 any the following (a) and/or (b): 30 (a) not less than 1.5 wt% to not more than 99 wt% of reduced coenzyme Q 9 relative to the reduced coenzyme Q10, and/or (b) reduced coenzyme Q11. 35 [2] The method of [1] , wherein the aforementioned (a) reduced coenzyme Q9 and (b) reduced coenzyme Q11 are separately prepared and added. [3] The method of [1], wherein the co-presence is achieved by reducing the oxidized coenzyme 10Q comprising oxidized coenzyme Q9 and/or oxidized coenzyme Q11.