a2) United States Patent
`US 9,028,877 B2
`(0) Patent No.:
`*May 12, 2015
`(45) Date of Patent:
`Bruheimet al.
`
`US009028877B2
`
`(54)
`
`(71)
`
`Applicant: Aker Biomarine Antarctic AS,
`Stamsund (NO)
`
`(72)
`
`Inventors:
`
`Inge Bruheim, Volda (NO); Snorre
`Tilseth, Bergen (NO); Daniele
`Mancinelli, Orsta (NO)
`
`(73)
`
`Assignee: Aker Biomarine Antarctic AS,
`Stamsund (NO)
`
`(*)
`
`Notice:
`
`Subject to any disclaimer, the term ofthis
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`
`This patent is subject to a terminal dis-
`claimer.
`
`(21)
`
`Appl. No.: 14/490,176
`
`(22)
`
`Filed:
`
`Sep. 18, 2014
`
`(65)
`
`(63)
`
`(60)
`
`Prior Publication Data
`
`US 2015/0004223 Al
`
`Jan. 1, 2015
`
`Related U.S. Application Data
`
`Continuation of application No. 12/057,775, filed on
`Mar. 28, 2008.
`
`Provisional application No. 60/920,483, filed on Mar.
`28, 2007, provisional application No. 60/975,058,
`filed on Sep. 25, 2007, provisional application No.
`60/983,446,
`filed on Oct. 29, 2007, provisional
`application No. 61/024,072,filed on Jan. 28, 2008.
`
`(51)
`
`Int. Cl.
`
`(52)
`
`(58)
`
`A6IK 45/06
`AGIK 31/23
`A6IK 31/122
`A6IK Y/48
`A6IK 31/683
`AG6IK 31/685
`C1IB 3/00
`A6IK 31/202
`US. Cl.
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`CPC wee A61K 35/612 (2013.01); A6LK 9/4858
`(2013.01); A6LK 31/122 (2013.01); A6LK
`31/23 (2013.01); A6LK 31/683 (2013.01);
`A6LK 31/685 (2013.01); A6LK 45/06
`(2013.01); CIB 3/006 (2013.01); A61K
`31/202 (2013.01)
`
`Field of Classification Search
`CPC . A61K 2300/00; A61K 31/122; A61K 31/23;
`A61K 31/683; A61K 35/612; A61K 41/0028;
`A61K 31/202; A61K 45/06; A61K 9/4858;
`A61K 47/48503; A61K 31/685; A61K
`2800/70; A61K 35/63; A61K 35/64; A61K
`8/553; A61K 8/925; A61K 35/60; A61K
`31/661
`
`See application file for complete search history.
`
`BIOEFFECTIVE KRILL OIL COMPOSITIONS
`
`(56)
`
`References Cited
`U.S. PATENT DOCUMENTS
`
`2,652,235 A
`4,036,993 A
`4,119,619 A
`4,133,077 A
`4,251,557 A
`4,505,936 A
`4,714,571 A
`4,749,522 A
`4,814,111 A
`5,006,281 A
`5,266,564 A
`5,434,183 A
`6,214,396 Bl
`6,346,276 Bl
`6,537,787 Bl
`6,800,299 BI
`7,488,503 Bl
`7,666,447 B2
`8,030,348 B2
`8,278,351 B2
`8,383,675 B2
`8,697,138 B2*
`2002/0076468 Al
`2003/0044495 Al
`2003/0113432 Al
`
`9/1953 Samuelsen
`T1977 Ikeda
`10/1978 Rogozhinet al.
`1/1979 Jasniewicz
`2/1981 Shimoseetal.
`3/1985 Meyerset al.
`12/1987 Kearnset al.
`6/1988 Kamarei
`3/1989 Kearnsetal.
`4/1991 Rubin etal.
`11/1993 Modolell
`7/1995 Larsson-Backstrom
`4/2001 Barrier
`2/2002 Tanouchiet al.
`3/2003 Breton
`10/2004 Beaudoinetal.
`2/2009 Porzioet al.
`2/2010 Rockway
`10/2011 Sampalis
`10/2012 Sampalis
`2/2013 Sampalis
`4/2014 Bruheimetal.
`6/2002 Saxby
`3/2003 Kagan etal.
`6/2003 Yoshitomi
`
`.....0.0.. 424/538
`
`(Continued)
`
`FOREIGN PATENT DOCUMENTS
`
`AU
`BR
`
`2002322233
`8701265
`
`2/2003
`3/1987
`
`(Continued)
`OTHER PUBLICATIONS
`
`Valeri, D., et al., “Visocities of Fatty acids, triglycerides and their
`binary mixtures,” JAOCS 74 (1997) pp. 1221-1226.
`CRC 2013-2014, 94th ed., pp. 6-231-6-235.
`Folch, et al., A Simple Method for the Isolation and Purification of
`Total Lipids from Animal Tissues. J. Biol. Chem., 226, 497-509
`(1957).
`Grant of Request for Ex parte Reexamination of the 351 patent.
`Grit et al., Hydrolysis of phosphatidylcholine in aqueous liposome
`dispersions, Int. J. Pharmaceutics 50:1-6 (1989).
`(Continued)
`
`Primary Examiner — Debbie K Ware
`(74) Attorney, Agent, or Firm — Casimir Jones S.C.
`
`(57)
`
`ABSTRACT
`
`This invention discloses new krill oil compositions charac-
`terized by having high amounts ofphospholipids, astaxanthin
`esters and/or omega-3 contents. The krill oils are obtained
`from krill meal using supercritical fluid extraction in a two
`stage process. Stage 1 removesthe neutrallipid by extracting
`with neatsupercritical CO, or CO,plus approximately 5% of
`a co-solvent. Stage 2 extracts the actual krill oils by using
`supercritical CO, in combination with approximately 20%
`ethanol. The krill oil materials obtained are compared with
`commercially available krill 011 and found to be more bioef-
`fective in a numberof areas such as anti-inflammation, anti-
`oxidant effects, improving insulin resistances and improving
`bloodlipid profile.
`
`19 Claims, 19 Drawing Sheets
`RIMFROST EXHIBIT 1068 Page 0001
`RIMFROST EXHIBIT 1068 Page 0001
`
`

`

`US 9,028,877 B2
`
`Page 2
`
`(56)
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`2004/0241249 Al
`2006/0078625 Al
`2006/0193962 Al
`2008/0166419 Al
`2008/0166420 Al
`2010/0143571 Al
`2010/0160659 Al
`2011/0130458 Al
`
`12/2004 Sampalis
`4/2006 Rockway
`8/2006 Kamiyaet al. oo... 426/615
`7/2008 Sones
`7/2008 Sones
`6/2010 Breivik
`6/2010 Catchpole
`6/2011 Breivik
`
`CA
`CA
`cL
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`GB
`GB
`JP
`JP
`a
`JP
`JP
`JP
`JP
`JP
`Ip
`Ip
`IP
`JP
`IP
`Jp
`JP
`JP
`JP
`ib
`ib
`JP
`JP
`IP
`IP
`JP
`JP
`JP
`JP
`JP
`JP
`a
`JP
`JP
`Ip
`IP
`JP
`SU
`WO
`
`FOREIGN PATENT DOCUMENTS
`1098900
`4/1981
`2251265
`4/2000
`40348
`7/1997
`0609078
`8/1994
`670306
`6/1995
`1127497
`8/2001
`1392623
`3/2004
`1406641
`4/2004
`1542670
`6/2005
`0973532
`9/2005
`1660071
`5/2006
`1689413
`8/2006
`1743531
`1/2007
`1631280
`3/2008
`1123368
`4/2008
`1406641
`1/2009
`1419768
`1/2009
`1292294
`3/2009
`1706106
`7/2009
`1385500
`7/2010
`2097014
`10/1982
`921537
`6/1999
`A-S51-125774
`11/1976
`A-S52-114046
`9/1977
`oeriao
`tose
`02049091
`2/1990
`2215351
`8/1990
`4012665
`1/1992
`2963152
`2/1992
`04057853
`2/1992
`3081692
`7/1994
`2524217
`8/1996
`HO8-231391
`9/1996
`3344887
`7/1997
`3611222
`8/1997
`2001-158736 A
`6/2001
`2003-003192 A
`1/2003
`2003-048831 A
`2/2003
`2003-146883 A
`5/2003
`5003ssoaae
`1soo,
`.
`2008RoR A
`ovoos
`2004-525180 A
`8/2004
`°
`2004-534800
`11/2004
`2004-536059 A
`12/2004
`3678317
`5/2005
`2005-245379 A
`9/2005
`2006-502196 A
`1/2006
`2006-069948 A
`3/2006
`2006-083136 A
`3/2006
`2006-290784 A
`10/2006
`2006-316073 A
`11/2006
`sooteon34 A
`oeoe
`.
`2007-502805 A
`2/2007
`2007-509131 A
`4/2007
`2007-126455 A
`5/2007
`2007-518764 A
`7/2007
`2007-246404 A
`9/2007
`220741
`1/1971
`86/06082
`10/1986
`
`WO
`WO
`wo
`WO
`we
`Wo
`wo
`Wo
`WO
`WO
`WO
`WO
`WO
`WO
`wo
`WO
`WO
`wo
`wo
`WO
`wo
`wo
`wo
`wo
`wo
`wo
`WO
`wo
`wo
`WO
`wo
`wo
`wo
`wo
`wo
`wo
`
`2/1989
`89/0103 1
`11/1989
`89/10960
`5/1990
`90/05765
`12/1993
`93/24142
`lovlooy
`ouneoe
`8/1998
`98/34498
`8/1999
`99/39589
`4/2000
`00/23546
`5/2000
`00/25608
`7/2000
`00/38708
`4/2001
`01/28526
`11/2001
`01/82928
`10/2002
`02/083 122
`11/2002
`02/092540
`12/2002
`02/102394
`2/2003
`03/011873
`2/2003
`03/013497
`4/2004
`2004/028529
`6/2004
`2004/047554
`11/2004
`2004/100943
`12/2004
`2004/112767
`1/2005
`2005/004393
`3/2005
`2005/018632
`4/2005
`2005/037848
`4/2005
`2005/038037
`8/2005
`2005/07041 1
`3/2006
`2006/030552
`10/2006
`2006/111633
`7/2007
`2007/080514
`7/2007
`2007/080515
`9/2007
`2007/108702
`11/2007
`2007/123424
`1/2008
`2008/006607
`6/2008
`2008/072563
`10/2008
`2008/117062
`3/2009
`2009/027692
`OTHER PUBLICATIONS
`
`Hendersonetal., Lipid Composition ofthe Pineal Organ from Rain-
`bow Trout (Oncorhynchus mykiss), Lipids, vol. 29, No. 5, pp. 311-
`317 (1994).
`:
`:
`Herman and Groves, The Influence of Free Fatty Acid Formation on
`ne
`vas
`:
`:
`.
`the pH of Phospholipid-Stabilized Triglyceride Emulsions, Pharma-
`:
`;
`ceutical Research 10(5):774-776 (1993).
`:
`:
`:
`Itano Refrigerated Food Co., Ltd, Bio & High Technology
`:
`:
`vas
`Announcement and Natural Astaxanthin & Krill Lecithin, pp. 1-16
`(on or before Dec. 28, 1994).
`Johnson and Lucas, Comparison of Alternative Solvents for Oils
`Extraction, JAOCS 60(2):229-242 (1983).
`;
`;
`Le Grandois et al., Investigation of Natural Phosphatidylholine
`Sources: Separation and Identification by Liquid Chromatography-
`Electrospray Ionization-Tandem Mass Spectrometry (LC-ESI-MS2)
`of Molecular Species, J. Agric. Food Chem., 57, 6014-20 (2009).
`Lin et al. Effect of Dietary N-3 Fatty Acids Upon the
`PhospholipidMolecular Species of the Monkey Retina,
`Invest
`Ophthalmol Vis Sci. 1994;35:794-803.
`Medinaet al., C Nuclear Magnetic Resonance Monitoring of Free
`Fatty Acid Release After Fish Thermal Processing, J. Amer. Oil
`Chem. Soc. 71(5):479-82 (1994).
`ffi
`Ty
`Oct. 24, 2012 Office Action, 675 patent.
`Office Action dated Jan. 5, 2012, °351 patent.
`U.S. Appl. No. 60/307,842 (Priority document for the ’351 patent).
`Supplemental Declaration of Bjorn Ole Haugsgjerd submitted during
`inter partes reexamination of parent patent U.S. 8,030,348.
`Supplemental Declaration of Dr. Earl White submitted duringinter
`partes reexamination of parent patent U.S. 8,030,348.
`Supplemental Declaration of Dr. Earl White submitted during pros-
`ecution ofparent patent U.S. 8,278,351.
`Supplemental Declaration of Dr. Thomas Gundersen submitted. dur-
`a
`oo.
`ing inter partes reexamination of parent patent U.S. 8,030,348.
`.
`:
`“
`vege
`:
`:
`Suzuki, T. and Shibata, N., The utilization of Antarctic krill for
`human food, Food Rev. Int 1, 6:1, 119-147 (1990).
`Takahashi et al., Compositional Changes in Molecular Species of
`Fish Muscle Phosphatidylcholine During Storage, Bull. Fac. Fish.
`Hokkaido Univ. 37(1), 80-84 1986.
`RIMFROST EXHIBIT 1068 Page 0002
`RIMFROST EXHIBIT 1068 Page 0002
`
`

`

`US 9,028,877 B2
`Page 3
`
`(56)
`
`References Cited
`OTHER PUBLICATIONS
`
`Takahashiet al., Molecular Species of Fish Muscle Lecithin, Bulletin
`of the Japanese Society of Scientific Fisheries 48(12), 1803-1814
`(1982).
`Takahashiet al., Prediction of Relative Retention Value of the Indi-
`vidual Molecular Species of Diacyl Glycerolipid on High Perfor-
`mance Liquid Chromatography, Bull. Fac. Fish. Hokkaido Univ.
`38(4), 398-404. 1987.
`1,2-dieicosapentaenoyl-sn-glycero-3-
`Tanaka, Biosynthesis of
`phosphocholine in Caenorhabditis elegans, Eur. J. Biochem. 263,
`189+194 (1999).
`Tocher, Chapter 6, Glycerophospholipid metabolism, Biochemistry
`and molecular biology offishes, vol. 4, Hochachka and Mommsen
`(eds.)(1995).
`Watanabeet al., Effective Components in Cuttlefish Meal and Raw
`Krill for Improvement of Quality of Red Seabream Pagrus major
`Eggs, Nippon Suisan Gakkaishi 57(4):68 1-694 (1991).
`WHONewsand Activities, Bulletin of the World Health Organiza-
`tion, 73(4), pp. 547-551 (1995).
`“Neptune Technologies & Bioressources Soon to Obtain a Major
`Patent in Over 30 Countries” (“2001 Press Release,”).
`Action Closing Prosecution, ’348 patent.
`Apr. 2, 2012 Response to Office Action, ’351 patent.
`Balassa et al., Microencapsulation in the Food Industry, Critical
`Reviewsin Food Technology, 2:2, 245-265 (1971).
`Bell and Dick, Molecular Species Composition of the Major Diacyl
`Glycerophospholipids from Muscle, Liver, Retina and Brain of Cod
`(Gadus morhua), Lipids, vol. 26, No. 8, pp. 565-573 (1991).
`Bell, Molecular Species Analysis of Phosphoglycerides from the
`Ripe Roesof Cod, Lipids, vol. 24, No. 7 (1989).
`Bell, Molecular Species Composition of Phosphatidylcholine from
`Crypthecodinium cohnii in Relation to Growth Temperature Lipids
`25, 115-118 (1990).
`Bergelson (ed.), Lipid Biochemical Preparations, Chapter 1.1, pp.
`1-13 (1980).
`Bottino, N.R., “Lipid Composition ofTwo Species ofAntarctic Krill:
`Euphausia superba and E. crystallorophias;’ Comp. Biochem.
`Physiol., 1975, vol. 50B, pp. 479-484.
`Buchi R-220 Rotovapor® Manual.
`and composition of
`Buda, Structural order of membranes
`phospholipids in fish brain cells during thermal acclimatization,
`Proc. Natl. Acad. Sci. USA vol. 91, pp. 8234-8238, Aug. 1994.
`Certificate of translation of Ex. 1072: Fisheries Agency, General
`Report on Research and Development of Techniques in Processing
`and Utilization of Marine Products, Chapter 6, Developmentof tech-
`nology for recovery of valuable substances (astaxanthin) from krill,
`by Takao Fujita, pp. 273-307 (Mar. 1985); Japanese language docu-
`ment.
`Japanese Patent No.
`Certificate of translation of Ex. 1074:
`60-153779, entitled “Nutritional Supplement”.
`Certificate of translation of Ex. 1076: Japanese Patent Publication
`No. H08-23 1391, entitled “Medicine for Improvement of Dementia
`Symptoms”.
`Certification oftranslation of Ex. 1070: Japanese Unexamined Patent
`Application Publication No. 02-215351.
`Certified translation of Ex. 1070: Japanese Unexamined Patent
`Application Publication No. 02-215351, titled Krill Phospholipids
`Fractioning Method (“Maruyama,”); Certificate of Translation pro-
`vided as Ex. 1071.
`Certified translation of Ex. 1072: Fisheries Agency, General Report
`on Research and Development of Techniques in Processing and Uti-
`lization of Marine Products, Chapter 6, Developmentof technology
`for recovery ofvaluable substances(astaxanthin) from krill, by Takao
`Fujita, pp. 273-307 (Mar. 1985) (“Fujita”) ; Certificate of Translation
`provided as Ex. 1073.
`Certified translation of Ex. 1074: Japanese Patent No. 60-153779,
`entitled “Nutritional Supplement” (“Fukuoka”); Certificate ofTrans-
`lation provided as Ex. 1075.
`
`Certified translation of Ex. 1076: Japanese Patent Publication No.
`H08-231391, entitled “Medicine for Improvement of Dementia
`Symptoms” (“Yasawa’); Certificate of Translation provided as Ex.
`1077.
`Declaration of Bjorn Ole Haugsgjerd in support of Inter Partes
`Review of U.S. Pat. No. 8,278,351.
`Declaration of Bjorn Ole Haugsgjerd submitted during inter partes
`reexamination of parent patent U.S. 8,030,348.
`Declaration of Dr. Albert Lee in Support of Inter Partes Review of
`USS. Pat. No. 8,278,351.
`Declaration of Dr. Albert Lee in Support of Inter Partes Review of
`USS. Pat. No. 8,383,675.
`Declaration of Dr. Chong Lee submitted during inter partes reexami-
`nation of parent patent U.S. 8,030,348.
`Declaration of Dr. Earl White submitted during prosecution ofparent
`patent U.S. 8,030,348.
`Declaration of Dr. Ivar Storre in support of Inter Partes Review of
`USS. Pat. No. 8,278,351.
`Declaration of Dr. Ivar Storre in support of Inter Partes Review of
`USS. Pat. No. 8,383,675.
`Declaration ofDr. Jacek Jaczynski from inter partes reexamination of
`the parent patent U.S. 8,030,348.
`Declaration of Dr. Jaczynski submitted. during prosecution of parent
`patent U.S. 8,278,351.
`Declaration of Dr. Jeff Moore in Support of Inter Partes Review of
`USS. Pat. No. 8,278,351.
`Declaration of Dr. Jeff Moore in Support of Inter Partes Review of
`USS. Pat. No. 8,383,675.
`Declaration of Dr. Richard van Breemen in Support of Inter Partes
`Review of U.S. Pat. No. 8,278,351.
`Declaration of Dr. Richard van Breemen in Support of Inter Partes
`Review of U.S. Pat. No. 8,383,675.
`Declaration of Dr. Shahidi submitted during inter partes reexamina-
`tion of parent patent U.S. 8,030,348.
`Declaration of Dr. Shahidi submitted during prosecution of parent
`patent U.S. 8,278,351.
`Declaration of Dr. Suzanne Budgein Support of Inter Partes Review
`of U.S. Pat. No. 8,278,351.
`Declaration of Dr. Suzanne Budgein Support of Inter Partes Review
`of U.S. Pat. No. 8,383,675.
`Declaration of Dr. Thomas Brennain support of Inter Partes Review
`of U.S. Pat. No. 8,278,351.
`Declaration of Dr. Thomas Brennain support of Inter Partes Review
`of U.S. Pat. No. 8,383,675.
`Declaration of Dr. Thomas Gundersen submitted during inter partes
`reexamination of parent patent U.S. 8,030,348.
`Declaration of Dr. Tina Sampalis submitted during inter partes reex-
`amination of parent patent U.S. 8,030,348.
`Declaration of Dr. Van Breemen submitted during Ex parte Reexami-
`nation of the ’351 patent.
`Declaration of Dr. Van Breemen submitted during Inter partes Reex-
`amination of the ’348 patent.
`Declaration of Dr. Yeboah submitted during inter partes reexamina-
`tion of parent patent U.S. 8,030,348.
`Declaration of Dr. Yeboah submitted during prosecution of parent
`patent U.S. 8,278,351.
`Eichberg, ““Lecithin—It Manufacture and Use in the Fat and Oil
`Industry,” Oils and Soap 51-54, 1939.
`Expert Witness Report ofDr. Theodore Welch submitted in relation to
`ITC Investigation No. 337-TA-877.
`Farkas, Composition and Physical State ofPhospholipids in Calanoid
`Copepodsfrom India and Norway, LIPIDS,vol. 23, No. 6 (1988).
`Final Prospectus dated May 11, 2001.
`Fisheries Agency, General Report on Research and Development of
`Techniquesin Processing and Utilization of Marine Products, Chap-
`ter 6, Development oftechnology for recovery ofvaluable substances
`(astaxanthin) from krill, by Takao Fujita, pp. 273-307 (Mar. 1985);
`Japanese language document.
`European Search Report, EP Patent Application No. EP12187516,
`mailed Jun. 10, 2013.
`CN Office Action mailed Apr. 27, 2012, JP Patent Application No.
`200880112125.6 (and English translation).
`RIMFROST EXHIBIT 1068 Page 0003
`RIMFROST EXHIBIT 1068 Page 0003
`
`

`

`US 9,028,877 B2
`
`Page 4
`
`(56)
`
`References Cited
`OTHER PUBLICATIONS
`
`Fricke,et al., Lipid, Sterol and Fatty Acid Composition of Antarctic
`Krill Euphausia superba Dana), Lipids (1984) 19(11): 821-827.
`Fricke,et al., 1-O-Alkylglycerolipids in Antarctic Krill (Euphausia
`superba Dana), Comp. Biochem.Physiol. (1986) 85B(1): 131-134.
`Gordeev, K.Y., et al. “Fatty Acid Composition of the Main
`Phospholipids of the Antarctic Krill, Euphausia superba,’ Chem.
`Nat. Cmpds. (1990) 26(2), pp. 143-147.
`Grantham (1977) Southern Ocean Fisheries Survey Programme,
`FAO Rome, GLO/SO/77/3: 1-61.
`Raventos et al., Application and Posssibilities of Supercritical CO2
`Extraction in Food Processing Industry: An Overview, Food Science
`and TechnologyInternational (2002) 8: 269-284.
`(PAF)-like
`Tanaka,
`T.,
`et
`al,
`Platelet-activating
`Factor
`Phospholoipds Formed during Peroxidation of Phosphatidylcholines
`from Different Foodstuffs, Biosci. Biotech. Biochem. (1995) 59 (8),
`pp. 1389-1393.
`Winther, et al., Elucidation of Phosphatidylcholine Composition in
`Krill Oil Extracted from Euphausia superba, Lipids (2011) 46:
`25-36.
`JP Office Action mailed Feb. 23, 2012, JP Patent Application No.
`2010-522444 (and English translation).
`Dec. 8, 2011 Office Action, KR Patent Application No. 10-2010-
`7006897 and its English translation.
`Sikorski, E., “The Utilization of Krill for Food,” Food Process Eng.,
`1:845-855 (1980).
`Budzinskli, E., et al., “Possibilities of processing and marketing of
`products made from Antarctic Krill”, FAO Fish. Tech. Pap. (268) 46
`pages (1985).
`BuneaR., et al.., “Evaluation of the Effects of Neptune Krill Oil on
`the Clinical Course of Hyperlipidemia,’ Alternative Medicine
`Review, Thorne Research Inc., Sandpoint, US, vol. 9, No. 4, Jan. 1,
`2004.
`Gordeev, K.Y., et al. “Fatty Acid Composition of the Main
`Phospholipids of the Antarctic Krill, Euphausia superba,’ Khim.
`Prirod. Soed. 2 (1990), pp. 181-187.
`Takaichiet al., 2003, “Fatty Acids of astaxanthinesters in krill deter-
`mined by mild mass spectrometry”, Comparative Biochemistry and
`Physiology Part B, Biochemistry and Molecular Biology, Elsevier,
`Oxford, vol. 136, Jan. 1, 2003, p. 317-322.
`Tanaka etal., 2004, “Extraction of Phospholipids from Salmon Roe
`with Supercritical Carbon Dioxide and an Entrainer”’, J. Oleo Sci,
`53(9): 417-424.
`Tanaka et al., 2005, “Extraction of Phospholipids from Unused
`Natrual Resources with Supercritical Carbon Dioxide and an
`Entrainer”’, Journal of Oleo Science, vol. 54(11): 569-576.
`Todoricet al., 2006, “Adiposetissue inflammation induced by high-
`fat diet in obese diabetic mice is prevented by n-3 polyunsaturated
`fatty acids”, Diabetologia, 49(9): 2109-2119.
`Touet al., 2007, “Krill for human consumption: nutritional value and.
`potential health benefits.”, Nutrition Rev 65(2):63-77.
`Trayhurnet al., 2004, “Adipokines: inflammation andthe pleiotropic
`role of white adipose tissue”, Br. J. Nutrition, 92(3): 347-355.
`Trebbleet al., 2003, “Inhibition of tumour necrosis factor-alpha and
`interleukin 6 production by mononuclear cells following dietary
`fish-oil supplementation in healthy men and responseto antioxidant
`co-supplementation”, Br. J. Nutrition, 90(2): 405-412.
`Ukkola et al., 2002, “Adiponectin: a link between excess adiposity
`and associated comorbidities?”, J. Mol. Med., 80(1 1): 696-702.
`Van Der Veenet al., 1971 “The Lipids of Krill (Euphausia Species)
`and Red Crab (Pleuroncodes planipes)y”’, Lipids, 6(7): 481-485.
`Virtue, et al. 1996, Reproductive trade-off in male Antarctic krill,
`Euphausia superba, Marine Biology, vol. 126, No. 3, pp. 521-527.
`Yamaguchiet al., 1983, “The Composition ofCarotenoid Pigments in
`the Antarctic Krill Euphausia superba’, Bulletin of the Japanese
`Society of Scientific Fisheries, 49(9): 1411-1415.
`Yamaguchiet al., 1986, “Supercritical Carbon Dioxide Extraction of
`Oils From Antarctic Krill,” Journal of Agricultural and Food Chem-
`istry, vol. 34, pp. 904-907.
`
`Yanase M; 1974, “Modification of a Russian methodfor separation of
`heat-coagulated protein from Antarctic krill”, Database FSTA
`(online); International Food Information Service (IFIS); Frankfurt-
`Main, DE.
`Yen et al., 1994, “Effect of dietary omega-3 and omega-6 fatty acid
`sources on PUVA-induced cutaneoustoxicity and tumorogenesis in
`the hairless mouse”, Arch. Dermatol. Res., 286(6): 331-6.
`Database WPI Week 200682, Thomson Scientific, London, GB,
`2006.
`English Abstract; JP 2003-531857; See abstract from corresponding
`WO 2001/082928filed herewith.
`English Abstract; JP 2004-525 180; See abstract from corresponding
`WO 2002/083 122filed herewith.
`English Abstract; JP 2006-528233; See abstract from corresponding
`WO 2004/100943 filed herewith.
`English Abstract; JP 2007-502805; See abstract from corresponding
`WO 2005/018632filed herewith.
`English Abstract; JP 2007-50913 1; See abstract from corresponding
`WO 2005/037848filed herewith.
`English Abstract; JP 2007-5 18764; See abstract from corresponding
`WO 2005/0704 11 filed herewith.
`English Abstract; JP 2004-536059; See abstract from corresponding
`WO 2002/09254filed herewith.
`English Abstract; JP 2006-502 196; See abstract from corresponding
`WO 2004/028529filed herewith.
`Ando and Hatano,
`1988, “Isolation of apolipoproteins from
`carotenoid-carrying lipoprotein in the serum of chum salmon,
`Oncorhynchus keta’’, J. Lipid Research, 29: 1264-1271.
`Aoi et al., 2003, “Astaxanthin limits exercise-induced skeletal and
`cardiac muscle damage in mice”, Antioxidants & Redox Signaling,
`5(1): 139-44.
`Britton, 1985, “General Carotenoid Methods”, Methodsin Enzymol-
`ogy, vol. 111, pp. 113-149.
`Calder, 2006, “‘n-3 polyunsaturated fatty acids, inflammation, and.
`inflammatory diseases”, Am. J. Clin. Nutr., 83: 1505S.
`Charestet al., 2001, “Astaxanthin Extraction from Crawfish Shells by
`Supercritical CO2 with Ethanol as Cosolvent”, J. Aquatic Food Prod-
`uct Technology, 10(3): 79-93.
`Chen and Meyers, 1982, “Extraction of Astaxanthin Pigment from
`Crawfish Waste Using a Soy Oil Process”, J. Food Sci., 47: 892-896.
`Clarke, 1980, “The Biochemical Composition of Krill, Euphausia
`superba dana,from South Georgia”, J. Exp. Mar. Biol. Ecol., 43:
`221-236.
`Czeczuga, 1974, “Comparative Studies of Carotenoids in the Fauna
`of the Gullmar Fjord (Bohuslan, Sweden). II. Crustacea: Eupagurus
`bernhardus, Hyas coarctatus and. Upogebia deltaura”, Marine Biol-
`ogy, 28: 95-98.
`De Ritter and Purcell, 1981, “Carotenoid Analytical Methods”,
`Carotenoids as Colorants and Vitamin A Precursors: Technological
`and Nutritional Applications, pp. 815-882.
`Deutch, 1995, “Menstrual pain in Danish womencorrelated with low
`n-3 polyunsaturated fatty acid intake”, Eur. J. Clin. Nutr., 49(7):
`508-16.
`Diez et al., 2003, “The role of the novel adipocyte-derived hormone
`adiponectin in human disease”, Eur. J. Endocrinol., 148(3): 293-300.
`Ellingsen et al., 1987, “Biochemistry of the autolytic processes in
`Antarctic krill post mortem. Autoproteolysis.” Biochem.J. 246, 295-
`305.
`Emodi, 1978, “Carotenoids: Properties and Applications”, Food.
`Technology, 32(5): 38.
`Felix-Valenzuela et al., 2001, “Supercritical CO2/Ethanol Extraction
`of Astaxanthin from Blue Crab (Callinectes sapidus) Shell Waste”,
`Journal of Food Process Engineering, 24: 101-112.
`Fox and Scheer, 1941, “Comparative Studies of the Pigments of
`Some Pacific Coast Echinoderms”, The Biological Bulletin, 441-
`455.
`Fricke, et al., 1984, “Lipid, Sterol and Fatty Acid Composition of
`Antarctic Krill (Euphausia superba Dana)”, Lipids, 19(11): 821-827.
`Geusenset al., 1994, “Long-term effect ofomega-3 fatty acid supple-
`mentation in active rheumatoid arthritis. A 12-month, double-blind,
`controlled study”, Arthritis Rheum., 37(6): 824-9.
`RIMFROST EXHIBIT 1068 Page 0004
`RIMFROST EXHIBIT 1068 Page 0004
`
`

`

`US 9,028,877 B2
`
`Page 5
`
`(56)
`
`References Cited
`OTHER PUBLICATIONS
`
`Gilchrist and Green, 1960, “The Pigments ofArtemia”, Proceedings
`ofthe Royal Society, Series B Biological Sciences, vol. 152 No. 946,
`pp. 118-136.
`Goodwin and Srisukh, 1949, “Some Observations on Astaxanthin
`Distribution in Marine Crustacea”, Department of Biochemistry,
`University of Liverpool, pp. 268-270.
`Gulyaev and Bugrova, 1976 “Removing fats from the protein paste
`Okean”. Konservnaya I Ovoshchesushil’naya Promyshlennost, (4),
`37-8.
`Hardardottir and Kinsella, 1988, “Extraction of Lipid and Choles-
`terol from Fish Muscle with Supercritical Fluids” Journal of Food
`Science, 53(6): 1656-1658.
`International Aqua Feed, 2006, vol. 9.
`International Search Report and Written Opinion for PCT/GB2008/
`002934, Dated Mar. 11, 2009.
`International Search Report and Written Opinion for PCT/IB2010/
`000512; dated Jun. 24, 2010.
`International Search Report for PCT/IB2007/000098, dated: Jun. 26,
`2007.
`Itoh et al., 2007; “Increased adiponectin secretion by highly purified.
`eicosapentaenoic acid in rodent models of obesity and human obese
`subjects”, Arteriosclerosis, Thrombosis, and Vascular Biology;
`27(9): 1918-1925.
`Johnsonet al., 1978, “Simple Methodfor the Isolation ofAstaxanthin
`from the Basidiomycetous Yeast Phaffia rhodozyma”, Applied and
`Environmental Microbiology, 35(6): 1155-1159.
`Kolakowska, 1989, “Krill lipids after frozen storage ofabout one year
`in relation to storage time before freezing”, Die Nahrung Food, 33(3):
`241-244.
`Kris-Etherton et al., 2002, “Fish Consumption, Fish Oil, Omega-3
`Fatty Acids, and Cardiovascular Disease”, Circulation, 106:2747-
`2757,
`Kristensen et al., 1989, “Dietary supplementation with n-3 polyun-
`saturated fatty acids and human platelet function: a review with
`particular emphasis on implications for cardiovascular disease”, J.
`Intern. Med. Suppl. 731:141-50.
`Kunesovaet al., 2006, “The influence of n-3 polyunsaturated fatty
`acids and very low calorie diet during a short-term weight reducing
`regimen on weight loss and serum fatty acid composition in severely
`obese women”, Physiol Res.; 55(1):63-72.
`Laightet al., 1999, “F2-isoprostane evidence of oxidantstress in the
`insulin resistant, obese Zucker rat: effects of vitamin E”, Eur. J.
`Pharmacol. 377(1): 89-92.
`“Method of Analysis of
`1971,
`Lambertson and Braekkan,
`Astaxanthin and its Occurrence in some Marine Products,” J. Sci.
`Food. Agr., vol. 22(2): 99-101.
`Libby et al., 2006, “Inflammation and Atherothrombosis: From
`Population Biology and Bench Research to Clinical Practice”, J.
`Amer. Coll. Card., 48 (9, Suppl. A): A33-A46.
`Lopezet al., 2004, “Selective extraction of astaxanthin from crusta-
`ceans by use of supercritical carbon dioxide”, Talanta, 64: 726-731.
`Mandeville, 1991, “Isolation and Identification of Carotenoid Pig-
`ments, Lipids and Flavor Active Components from Raw Commercial
`Shrimp Waste”, Food Biotechnology, 5(2): 185-195.
`Meyers and Bligh, 1981, “Characterization of Astaxanthin Pigments
`from Heat-Processed Crawfish Waste”, J. Agric. Food Chem., 29:
`505-508.
`Meyers, 1977, “Using Crustacean Meals and Carotenoid-Fortified
`Diets”, Feedstuffs, vol. 49(19).
`Meyers, 1994, “Developments in world aquaculture, feed. formula-
`tions, and role of carotenoids”, Pure & Appl. Chem, vol. 66(5):
`1069-1076.
`
`Mills et al., 1989, “Dietary N-6 and N-3 fatty acids andsalt-induced.
`hypertension in the borderline hypertensive rat”, Lipids, 24(1):
`17-24.
`Moates and Van Bentem, 1990, “Separating out the value”, Food.
`Science and Technology Today, 4(4): 213-214.
`Nikolaeva, 1967 “Amino acid composition of protein-coagulate in
`lvill”, VNIRO, 63:161-4.
`Phleger,et al. (2002) “Interannual and between species comparison
`in the lipids, fatty acids, and sterols of Antarctic krill from the US
`AMLR Elephant Island survey area: 1997 and 1998”. Comp
`Biochem Physiol 131B:733-747.
`Popp-Snijders et al., 1987, “Dietary supplementation of omega-3
`polyunsaturated fatty acids improves insulin sensitivity in non-insu-
`lin-dependent diabetes”, Diabetes Res. 4(3): 141-7.
`Sachindra, 2006, “Recovery of carotenoids from shrimp waste in
`organic solvents”, Waste Management, 26: 1092-1098.
`Saether et al., 1986, “Lipids of North Atlantic krill”, J Lipid Res.,
`27(3):274-85.
`Shahidi et al., 1998, “Carotenoid Pigments in Seafoods and
`Aquaculture” Critical Reviews in Food Science, 38(1): 1-67.
`Sidehuet al., 1970, “Biochmical Composition and Nutritive Value of
`Krilluphausia superb dana)’, J. Sci Food Agr., vol. 21, 293-296.
`Simopoulos, 1991, “Omega-3 fatty acidsin health and disease and in
`growth and development”, Am. Clin. Nutr. 54:438-63.
`Somiya, 1982, “‘Yellow lens’ eyes of a stomiatoid deep-sea fish,
`Malacosteus niger”, Proc. R. Soc. Lond., 215: 481-489.
`EP Oppositionfiled Feb. 13, 2014 by Olympic Seafood AS, EP Patent
`Application No. EP08718910I6.
`Brzustowicz, Michael R., et al., “Controlling Membrane Cholesterol
`Content. A Role
`for Polyunsaturated
`(Docosahexaenoate)
`Phospholipids,” Biochemistry (2002), 41, pp. 12509-12519.
`Jong-HoLee, “A Review: Antioxygenic and Peroxide-decomposing
`Activities ofAntarctic Krill Lipids,” J. Korean Soc. Food Mutr. 13(3)
`pp. 326-333 (1984).
`Ki Woong Cho, et al., “Lipid and Fatty Acid Composition of the
`Antarctic Krill Euphausia superba,’ Ocean Research 21(2): 109-116
`(1999).
`Hvattum, Erlend, et al., “Effect of soybean oil and fish oil on indi-
`vidual molecular species of Atlantic salmon ...”, Journal of Chro-
`matography B, 748 (2000) 137-149.
`Igarashi, Daisuke,et al., “Positional Distribution of DHA and EPAin
`Phosphatidylcholine and Phosphatidylethanolamine from Different
`Tissues of Squids,” J. Oleo Sci. vol. 50, No. 9 (2001).
`Tochizawa, Kaoru, et al., “Effects of Phospholipds Containing
`Docosahexaenoic Acid on Differentiation and Growth of HL-60
`Human Promyelocytic Leukemia Cells,” J. Jpn. Oil Chem. Soc. vol.
`46, No. 4 (1997).
`Zerouga, Mustapha, et al., “Comparison of phosphatidylcholines
`containing one or two docosahexaenoic acyl chains on properties of
`phospholipid monolayers and bilayers,” Biochimica et Biophysica
`Acta 1236 (1995) 266-272.
`Eung-HoLee,et al., “Studies on the Processing of Krill Sauce,” J.
`Korean Soc. Food Nutr. 13(1) 97-106 (1984).
`Hyun-Ku Kim,et al., “Effects of Cooking and Drying Methodson the
`Polar Lipds Composition of Shrimp,” Korean J. Food Sci. Technol.
`vol. 21, No. 1, pp. 25-30 (1989).
`Shon, Mi-Yae,et al., “Effects of Krill and Cadmium on Lipid Com-
`position of Plasma in Cholesterol-Fed. Rats,’ J. Korean Soc. Food
`Nutr. 23(1), 38-43 (1994).
`SummonsMaterials downloaded from ESPACEon Dec. 16, 2014 for
`EP Patent Application No. 08 718 910.6.
`
`* cited by examiner
`
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`et
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`
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`Chol
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`Fig. 3
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`Sheet 4 of 19
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`Fig. 4
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`Insulin,
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`microU/L
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`RIMFROST EXHIBIT 1068 Page 0010
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`Sheet 6 of 19
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`US 9,028,877 B2
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`Fig. 6
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`
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`Sheet 11 of 19
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`RIMFROST EXHIBIT 1068 Page 0016
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`Fig. 12
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`(grams)
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`Sheet