(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2011/0104297 A1
`BRUHEIM et al.
`(43) Pub. Date:
`May 5, 2011
`
`US 201 l0l04297A1
`
`(54) METHODS OF USING KRILL OIL TO TREAT
`RISK FACTORS FOR CARDIOVASCULAR,
`METABOLIC, AND INFLAMMATORY
`DISORDERS
`
`61/024,072, filed on Jan. 28, 2008, provisional appli-
`cation No. 61/181,743, filed on May 28, 2009, provi-
`sional application No. 60/920,483, filed on Mar. 28,
`2007.
`
`(75)
`
`Inventors:
`
`Inge BRUHEIM, Volda (NO);
`Snorre Tilseth, Bergen (NO);
`Jeffery Cohn, Sydney (AU);
`Mikko Griinari, Espoo (Fl);
`Daniele Mancinelli, Orsta (NO);
`Nils Hoem, Oslo (NO); Hogne Vik,
`Eiksmarka (NO); Sebastiano
`Banni, Calgliari (IT)
`
`(73) Assignee:
`
`Aker BioMarine A.S.A., Oslo
`(N0)
`
`(21) Appl. No.:
`
`12/790,575
`
`(22)
`
`Filed:
`
`May 28, 2010
`
`Related U.S. Application Data
`
`(63) Continuation-in-part of application No. 12/057,775,
`filed on Mar. 28, 2008.
`
`(60) 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.
`
`Publication Classification
`
`(51)
`
`Int. Cl.
`(2006.01)
`A61K 35/56
`(2006.01)
`A61P 9/10
`(2006.01)
`A61P 3/04
`(2006.01)
`A61P 3/00
`(52) U.S. Cl. ........................................... .. 424/522; 426/2
`
`(57)
`
`ABSTRACT
`
`This invention discloses methods of using krill oil and com-
`positions comprising krill oil to treat risk factors for meta-
`bolic, cardiovascular, and inflammatory disorders. The
`present invention also relates to methods of using composi-
`tions comprising krill oil to modulate biological processes
`selected from the group consisting of glucose metabolism,
`lipid biosynthesis, fatty acid metabolism, cholesterol biosyn-
`thesis, and the mitochondrial respiratory chain. The present
`invention further includes pharmaceutical and/or nutraceuti-
`cal formulations made from krill oil, methods ofmaking such
`formulations, and methods of administering them to treat risk
`factors for metabolic, cardiovascular, and inflammatory dis-
`orders.
`
`NEPN 2007
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`Patent Application Publication
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`May 5, 2011 Sheet 1 of 26
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`US 2011/0104297 A1
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`May 5,2011
`
`METHODS OF USING KRILL OIL TO TREAT
`RISK FACTORS FOR CARDIOVASCULAR,
`METABOLIC, AND INFLAMMATORY
`DISORDERS
`
`[0001] This application claims the benefit of U.S. Prov.
`Appl. 61/181,743, filed May 28, 2009, and is a continuation-
`in-part ofU.S. application Ser. No. 12/057,775, filed Mar. 28,
`2008, which claims the benefit of U.S. Prov. Appl. 60/920,
`483, filed Mar. 28, 2007, U.S. Prov. Appl. 60/975,058, filed
`Sep. 25, 2007, U.S. Prov. Appl. 60/983,446, filed Oct. 29,
`2007, and U.S. Prov. Appl. No. 61/024,072, filed Jan. 28,
`2008, all of which are incorporated by reference herein in
`their entirety.
`
`FIELD OF THE INVENTION
`
`[0002] This invention relates generally to methods ofusing
`krill oil to treat risk factors for metabolic, cardiovascular, and
`inflammatory disorders, including, but not limited to, modu-
`lating endocarmabinoid concentrations; reducing ectopic fat;
`reducing triacylglycerides in the liver and heart; reducing
`monoacylglyceride lipase activity in the visceral adipose tis-
`sue, liver, and heart; increasing levels of DHA in the liver;
`increasing the levels of EPA and DHA in the phospholipid
`fractions of tissues that exhibit changes in endocannabinoid
`concentration;
`reducing susceptibility to inflammation,
`modulating glucose and lipid homeostasis; reducing fatty
`liver disease (alcoholic and non-alcoholic); reducing MAGL
`activity in the heart; increasing levels of plasma ALA/LA;
`decreasing levels of ALA/LA in the heart; decreasing levels
`of ARA in the subcutaneous adipose tissue; and decreasing
`availability of substrates to decrease the activity of the
`endocannabinoid system. The present invention also relates
`to methods of using compositions comprising krill oil to
`modulate biological processes selected from the group con-
`sisting of glucose metabolism, lipid biosynthesis, fatty acid
`metabolism, cholesterol biosynthesis, and the mitochondrial
`respiratory chain. The present invention further includes
`pharmaceutical and/or nutraceutical formulations made from
`krill oil, methods of making such formulations, and methods
`of administering them to treat risk factors for metabolic,
`cardiovascular, and inflammatory disorders.
`
`BACKGROUND OF THE INVENTION
`
`[0003] Krill is a small crustacean which lives in all the
`major oceans worldwide. For example, it can be found in the
`Pacific Ocean (Euphausia paczfica), in the Northern Atlantic
`(Meganycziphanes norvegica) and in the Southern Ocean off
`the coast of Antarctica (Euphausia superba). Krill is a key
`species in the ocean as it is the food source for many animals
`such as fish, birds, sharks and whales. Krill can be found in
`large quantities in the ocean and the total biomass ofAntarctic
`krill (Euphausia superba) is estimated to be in the range of
`300-500 million metric tons. Antarctic krill feeds on phy-
`toplankton during the short Antarctic summer. During winter,
`however, its food supply is limited to ice algae, bacteria,
`marine detritus as well as depleting body protein for energy.
`Virtue et al., Mar Biol. 126, 521-527. For this reason, the
`nutritional values of krill vary during the season and to some
`extent armually. Phleger et al., Comp. Biochem. Physiol. 131
`B (2002) 733. In order to accommodate variations in food
`supply, krill has developed an efficient enzymatic digestive
`
`apparatus resulting in a rapid breakdown of the proteins into
`amino acids. Ellingsen et al., Biochem. J. (1987) 246, 295-
`305. This autoproteolysis is highly eflicient also postmortem,
`making it a challenge to catch and store the krill in a way that
`preserves the nutritional quality of the krill. Therefore, in
`order to prevent the degradation ofkrill the enzymatic activity
`is either reduced by storing the krill at low temperatures or the
`krill is made into a krill meal.
`
`[0004] During the krill meal process the krill is cooked so
`that all the active enzymes are denatured in order to eliminate
`all enzymatic activity. Krill is rich in phospholipids which act
`as emulsifiers. Thus, it is more diflicult to separate water, fat,
`and proteins using mechanical separation methods than it is in
`a regular fish meal production line. In addition, krill becomes
`solid, gains weight and loses liquid more easily when mixed
`with hot water. Eventually this may lead to a gradual build up
`of coagulated krill proteins in the cooker and a non-continu-
`ous operation due to severe clogging problems. In order to
`alleviate this, hot steam must be added directly into the
`cooker. This operation is energy demanding and may also
`result in a degradation of unstable bioactive components in
`the krill oil, such as omega-3 fatty acids, phospholipids and
`astaxanthin. The presence of these compounds make krill oil
`an attractive source as a food supplement, a functional food
`product, and a pharmaceutical for the animal and human
`applications.
`[0005] Omega-3 fatty acids have been shown to have poten-
`tial effect ofpreventing cardiovascular disease, cognitive dis-
`orders, joint disease and inflammation-related diseases such
`as rheumatoid arthritis and osteoarthritis. Astaxanthin is a
`
`strong antioxidant and may also assist in promoting optimal
`health.
`
`Published PCT Application No. WO 00/23546 dis-
`[0006]
`closes isolation of krill oil from krill using solvent extraction
`methods. Krill lipids have been extracted by placing the mate-
`rial in a ketone solvent (e.g., acetone) in order to extract the
`lipid soluble fraction. This method involves separating the
`liquid and solid contents and recovering a lipid rich fraction
`from the liquid fraction by evaporation. Further processing
`steps include extracting and recovering by evaporation the
`remaining soluble lipid fraction from the solid contents by
`using a solvent such as ethanol. The compositions produced
`by these methods are characterized by containing at least 75
`pg/g astaxanthin, preferably 90 pg/g astaxanthin. Another
`krill lipid extract disclosed contained at least 250 ug/g canas-
`taxanthin, preferably 270 ug/g canastaxanthin.
`[0007]
`Published PCTApplication No. WO 02/102394 dis-
`closes methods of treating and/or preventing cardiovascular
`disease, rheumatoid arthritis, skin cancer, premenstrual syn-
`drome, diabetes, and enhancing transdermal transport. The
`methods include administering a krill or marine oil to a
`patient. The application also describes a test that was carried
`out to evaluate the effects of krill and/or marine oils on arte-
`
`riosclerotic coronary artery disease and hyperlipidemia, and
`resulted in a cholesterol decrease of about 15%, a triglyceride
`decrease of about 15%, an HDL increase of about 8%, an
`LDL decrease of about 13%, and a cholesterol:HDL ratio
`decrease of about 14%
`
`Published PCT Application No. WO 2007/080515
`[0008]
`discloses a marine lipid extract derived from krill. The extract
`can be used in methods for preventing or treating thrombosis.
`[0009] Korean PublishedApplication No. 2006008155 dis-
`closes an oral composition comprising a mixture of glu-
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`cosamine and krill oil (provided in a ratio of 2:3) for use in
`methods of inhibiting osteoarthritis.
`[0010] U.S. Pat. No. 7,666,447 discloses compositions
`including krill extracts and conjugated linoleic acid. The
`compositions are used in methods for treating an individual
`having a disease state selected from the group consisting of a
`joint ailment, PMS, Syndrome X, cardiovascular disease,
`bone disease and diabetes. The methods comprise adminis-
`tering to the individual a therapeutically effective amount of
`a composition including conjugated linoleic acid and a krill
`extract comprising krill oil.
`[001 1] However, there remains a need in the art for methods
`ofusing compositions comprising krill oil to treat risk factors
`for metabolic, cardiovascular, and inflammatory disorders.
`
`SUMMARY OF THE INVENTION
`
`[0012] The present invention provides methods of using
`compositions comprising krill oil (KO) to treat risk factors for
`metabolic, cardiovascular, and inflammatory disorders,
`including, but not limited to, modulating endocannabinoid
`concentrations; reducing ectopic fat; reducing triacylglycer-
`ides in the liver and heart; reducing monoacylglyceride lipase
`activity in the visceral adipose tissue, liver, and heart; increas-
`ing levels of DHA in the liver; increasing the levels of EPA
`and DHA in the phospholipid fractions of tissues that exhibit
`changes in endocannabinoid concentration; reducing suscep-
`tibility to inflammation, modulating glucose and lipid
`homeostasis; reducing fatty liver disease (alcoholic and non-
`alcoholic); reducing MAGL activity in the heart; increasing
`levels of plasma ALA/LA; decreasing levels of ALA/LA in
`the heart; decreasing levels ofARA in the subcutaneous adi-
`pose tissue; and decreasing availability of substrates to
`decrease the activity of the endocannabinoid system. The
`present invention also provides methods of using composi-
`tions comprising krill oil to modulate biological processes
`selected from the group consisting of glucose metabolism,
`lipid biosynthesis, fatty acid metabolism, cholesterol biosyn-
`thesis, and the mitochondrial respiratory chain. The present
`invention further includes pharmaceutical and/or nutraceuti-
`cal formulations made from the compositions, methods of
`making such formulations, and methods of administering
`them to treat risk factors for metabolic, cardiovascular, and
`inflammatory disorders.
`[0013]
`In some embodiments, the present invention pro-
`vides methods of administering compositions comprising
`krill oil to treat risk factors for metabolic, cardiovascular, and
`inflammatory disorders in a human subject, where the method
`includes the step of administering compositions containing
`krill oil. The risk factors that are treated are selected from the
`
`group consisting of modulating endocannabinoid concentra-
`tions; reducing ectopic fat; reducing triacylglycerides in the
`liver and heart; reducing monoacylglyceride lipase activity in
`the visceral adipose tissue, liver, and heart; increasing levels
`of DHA in the liver; increasing the levels of EPA and DHA in
`the phospholipid fractions of tissues that exhibit changes in
`endocannabinoid concentration; reducing susceptibility to
`inflammation, modulating glucose and lipid homeostasis;
`reducing fatty liver disease (alcoholic and non-alcoholic);
`reducing MAGL activity in the heart; increasing levels of
`plasma ALA/LA; decreasing levels of ALA/LA in the heart;
`decreasing levels ofARA in the subcutaneous adipose tissue;
`and decreasing availability of substrates to decrease the activ-
`ity of the endocannabinoid system.
`
`In some embodiments, the present invention pro-
`[0014]
`vides methods of administering compositions comprising
`krill oil to modulate biological processes in a human subject,
`where the method includes the step of administering compo-
`sitions containing krill oil. The biological processes are
`selected from the group consisting of glucose metabolism,
`lipid biosynthesis, fatty acid metabolism, cholesterol biosyn-
`thesis, and the mitochondrial respiratory chain. These bio-
`logical processes may be modulated by altering the expres-
`sion of one or more genes, including, but not limited to,
`reduced or decreased expression of Ppargcla (peroxisome
`proliferator-activated receptor gamma coactivator la), Hnf4a
`(hepatocyte nuclear factor 4 alpha), Pckl (phosphoenolpyru-
`vate carboxykinase 1), G6 pc (glucose-6-phosphatase, cata-
`lytic), Cptla (camitine palmitoyl
`transferase la), Acads
`(acyl-coenzyme A dehydrogenase, short chain), Acadm
`(acyl-coenzyme A dehydrogenase, medium chain), Acadl
`(acyl-coenzymeA dehydrogenase, long chain), Hmgcr (3 -hy-
`droxy-3-methylglutaryl-coenzyme A reductase), Pmvk
`(phosphomevalonate kinase), Sbref2 (sterol regulatory ele-
`ment binding factor 2), Ppargclb (peroxisome proliferator-
`activated receptor gamma coactivator lb), and Sod2 (super-
`oxide dismutase 2). These biological processes may also be
`affected by enhanced or increased expression of NADH
`(nicotinamide adenine dinucleotide) dehydrogenase and sub-
`units thereof. The biological processes are also affected by
`factors
`including reduced hepatic glucose production,
`reduced hepatic gluconeogenesis, and reduced hepatic lipid
`synthesis.
`[0015]
`In some embodiments, the present invention pro-
`vides methods of decreasing lipid content in the liver of a
`human subject, comprising: administering to said subject an
`effective amount of a krill oil composition under conditions
`such that lipid content in the liver of the subject is decreased.
`In some embodiments, the human subject is clinically obese.
`[0016]
`In certain embodiments, the present invention pro-
`vides methods comprising providing a krill oil composition to
`a human subject under conditions such that the cardiovascular
`disease risk factors of the subject are improved. In some
`embodiments, the cardiovascular risk factors are selected
`from the group consisting of elevated blood pressure,
`elevated serum total cholesterol and low-density lipoprotein
`cholesterol (LDL-C),
`low serum high-density lipoprotein
`cholesterol (HDL-C), diabetes mellitus, abdominal obesity,
`elevated serum triglycerides, small LDL particles, elevated
`serum homocysteine, elevated serum lipoprotein(a), pro-
`thrombotic factors, fatty liver and inflammatory markers. In
`some embodiments, the human subject is clinically obese.
`[0017]
`In certain embodiments, the present invention pro-
`vides methods comprising providing a krill oil composition to
`a human subject under conditions such that carmabinoid
`receptor signaling is reduced. In some embodiments, inhibi-
`tion of the endocannabinoid system of the subject comprises
`lowering the levels of arachidonylethanolarnide (AEA) and/
`or 2-arachidonyl glycerol (2-AG). In some embodiments, the
`human subject is clinically obese.
`[0018]
`In certain embodiments, the present invention pro-
`vides methods comprising providing a krill oil composition to
`a human subject; and administering the krill oil composition
`to the human subject under conditions such that the appetite
`of the subject is reduced. In some embodiments, the human
`subject is clinically obese.
`[0019]
`In certain embodiments, the present invention pro-
`vides methods comprising providing a krill oil composition to
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`a human subject; and administering the krill oil composition
`to the human subject under conditions such that fat accumu-
`lation in the subject is reduced. In some embodiments, the
`human subject is clinically obese.
`[0020]
`In certain embodiments, the present invention pro-
`vides uses of a krill oil composition in a human subject for
`improvement of cardiovascular disease risk factors, reduction
`of cannabinoid receptor signaling, reduction of appetite,
`reduction of fatty heart or reduction of fat accumulation.
`[0021]
`In certain embodiments, the present invention pro-
`vides uses of krill oil for the preparation of a medicament for
`improvement of cardiovascular disease risk factors, reduction
`in cannabinoid receptor signaling, reduction of appetite, or
`reduction of fat accumulation.
`
`In certain embodiments, the present invention pro-
`[0022]
`vides methods comprising providing a krill oil composition to
`a human subject; and administering the krill oil composition
`to the human subject under conditions such that the reproduc-
`tive performance is increased. In some embodiments, repro-
`ductive performance is improved chance of ovulation in
`females. In some embodiments, reproductive performance is
`spermatogenesis, sperm motility and/or acreosome reaction.
`[0023]
`In certain embodiments, the present invention pro-
`vides methods comprising providing a krill oil composition to
`a human subject; and administering the krill oil composition
`to the human subject under conditions such the liver and/or
`kidney functions are improved.
`[0024] Other novel features and advantages of the present
`invention will become apparent to those skilled in the art upon
`examination of the following or upon learning by practice of
`the invention.
`
`DETAILED DESCRIPTION OF THE DRAWINGS
`
`FIG. 1. 31 P NMR analysis ofpolar lipids in krill oil.
`[0025]
`FIG. 2. Blood lipid profiles in Zucker rats fed dif-
`[0026]
`ferent forms of omega-3 fatty acids (TAG:FO, PLl:NKO
`and PL2:Superba).
`[0027]
`FIG. 3. Plasma glucose concentration in Zucker rats
`fed different forms of omega-3 fatty acids.
`[0028]
`FIG. 4. Plasma insulin concentration in Zucker rats
`fed different forms of omega-3 fatty acids.
`[0029]
`FIG. 5. Estimated HOMA-IR values in Zucker rats
`fed different forms of omega-3 fatty acids.
`[0030]
`FIG. 6. The effect of dietary omega-3 fatty acids on
`TNF-a production by peritoneal macrophages.
`[0031]
`FIG. 7. The effect of dietary omega-3 fatty acids on
`lipid accumulation in the liver.
`[0032]
`FIG. 8. The effect of dietary omega-3 fatty acids on
`lipid accumulation in the muscle.
`[0033]
`FIG. 9. The effect of dietary omega-3 fatty acids on
`lipid accumulation in the heart.
`[0034]
`FIG. 10. Relative concentrations of DHA in the
`brain in Zucker rats supplemented with omega-3 fatty acids.
`[0035]
`FIG. 11. Mean group body weights (g) in the col-
`lagen-induced male DBA/1 arthritic mice. B-PL2 is the krill
`oil group. * p<0.05, significantly different from Group A
`(Positive Control—Fish Oil) and Group C (Control).
`[0036]
`FIG. 12. Body weight for the various treatment
`groups.
`[0037]
`groups.
`FIG. 14. Muscle to body weight ratio for the various
`[0038]
`treatment groups.
`
`FIG. 13. Muscle weight for the various treatment
`
`FIG. 15. Serum adiponectin levels (ng/ml) for the
`[0039]
`various treatment groups.
`[0040]
`FIG. 16. Serum insulin levels for the various treat-
`ment groups.
`[0041]
`FIG. 17. Blood glucose (mmol/l) levels in the vari-
`ous treatment groups.
`[0042]
`FIG. 18. HOMA-IR values for the various treatment
`groups.
`
`FIG. 19. Liver triglyceride levels (p.mol/g) for the
`[0043]
`various treatment groups.
`[0044]
`FIG. 20A-B. Levels of anandarnide (arachidonoyl
`ethanolamide) and 2-arachidonoyl glycerol in visceral adi-
`pose tissue in Zucker rats.
`[0045]
`FIG. 21A-B. Levels of anandarnide (arachidonoyl
`ethanolamide) and 2-arachidonoyl glycerol in subcutaneous
`adipose tissue in Zucker rats.
`[0046]
`FIG. 22A-B. Levels of anandarnide (arachidonoyl
`ethanolamide) and 2-arachidonoyl glycerol in liver tissue in
`Zucker rats.
`
`FIG. 23A-B. Levels of anandarnide (arachidonoyl
`[0047]
`ethanolamide) and 2-arachidonoyl glycerol in heart tissue in
`Zucker rats.
`
`FIG. 24. Triacylglyceride content in liver.
`[0048]
`FIG. 25. Triacylglyceride content in heart.
`[0049]
`FIG. 26. Cholesterol profile in plasma.
`[0050]
`FIG. 27. Fatty acid analyses of monocytes.
`[0051]
`FIG. 28. TNF alpha release in peritoneal monocytes
`[0052]
`after ex vivo challenge with LPS.
`[0053]
`FIG. 29A-B. Liver (A) and heart (B) triacylglycerol
`concentrations of obese Zucker rats fed control, fish oil, or
`krill oil diets for four weeks. Values are expressed as mean +/—
`SD, n:6. Means that do not have a common letter differ,
`P<0.05.
`
`FIG. 30A-B. Visceral AEA (A) and 2-AG (B) con-
`[0054]
`centrations in obese Zucker rats fed control, fish oil, or krill
`oil diets for four weeks. Values are expressed as mean +/— SD,
`n:6. Means that do not have a common letter differ, P<0.05.
`[0055]
`FIG. 31A-D. Liver (A and B) and heart (C and D)
`AEA (A and C) and 2-AG (B and D) concentrations in rats fed
`control, fish oil, or krill oil diets for four weeks. Values are
`expressed as mean +/— SD, n:6. Means that do not have a
`common number differ, P<0.05.
`[0056]
`FIG. 32A-B. Cholesterol (A) and TAG (B) concen-
`trations in plasma from rats fed control (C), fish oil (FO), or
`krill oil (KO) diets. Error bars depict S.D., n:6. Different
`letters denote significant differences (p<0.05)
`[0057]
`FIG. 33. Treatment-induced changes in the expres-
`sion of the mitochondrial reactive oxygen species detoxifica-
`tion enzyme Sod2. Expression was significantly decreased by
`a KO diet.
`
`suggesting decreased glucose
`FIG. 34. Genes
`[0058]
`uptake and increased fructose metabolism. KO diet showed a
`trend for increased Aldob expression (p:0.022)
`[0059]
`FIG. 35. Key genes regulating hepatic glucose pro-
`duction
`
`[0060]
`lism.
`
`FIG. 36. Key genes involved in fatty acid metabo-
`
`FIG. 37. Key genes regulating cholesterol biosyn-
`[0061]
`thesis in the liver 3-hydroxy-3-methylglutaryl-Coenzyme A
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`FIG. 38. Transcriptional cofactors and gene targets
`[0062]
`proposed to mediate the effect of krill-supplements on
`hepatic glucose metabolism and lipid biosynthesis.
`
`DEFINITIONS
`
`[0063] An “ether phospholipid” as used herein preferably
`refers to a phospholipid having an ether bond at position 1 of
`the glycerol backbone. Examples of ether phospholipids
`include, but are not limited to, alkylacylphosphatidylcholine
`(AAPC), lyso-alkylacylphosphatidylcholine (LAAPC), and
`alkylacylphosphatidylethanolarnine (AAPE). A “non-ether
`phospholipid” is a phospholipid that does not have an ether
`bond at position 1 of the glycerol backbone.
`[0064] As used herein, the term “omega-3 fatty acid” refers
`to polyunsaturated fatty acids that have the final double bond
`in the hydrocarbon chain between the third and fourth carbon
`atoms from the methyl end of the molecule. Non-limiting
`examples of omega-3 fatty acids include, 5,8,11,14,17-
`eicosapentaenoic
`acid
`(EPA),
`4,7,10,13,16,19-docosa-
`hexaenoic acid (DHA) and 7,10,13,16,19-docosapentaenoic
`acid (DPA).
`[0065] As used herein, the term “W/W (weight/weight)”
`refers to the amount of a given substance in a composition on
`weight basis. For example, a composition comprising 50%
`W/W phospholipids means that the mass of the phospholipids
`is 50% ofthe total mass ofthe composition (i.e., 50 grams of
`phospholipids in 100 grams of the composition, such as an
`oil).
`
`DETAILED DESCRIPTION OF THE INVENTION
`
`[0066] The present invention relates to methods of using
`krill oil and/or compositions comprising krill oil to treat risk
`factors for metabolic, cardiovascular, and inflammatory dis-
`orders, including, but not limited to, modulating endocannab-
`inoid concentrations; reducing ectopic fat; reducing triacylg-
`lycerides in the liver and heart; reducing monoacylglyceride
`lipase activity in the visceral adipose tissue, liver, and heart;
`increasing levels of DHA in the liver; increasing the levels of
`EPA and DHA in the phospholipid fractions of tissues that
`exhibit changes in endocannabinoid concentration; reducing
`susceptibility to inflammation, modulating glucose and lipid
`homeostasis; reducing fatty liver disease (alcoholic and non-
`alcoholic); reducing MAGL activity in the heart; increasing
`levels of plasma ALA/LA; decreasing levels of ALA/LA in
`the heart; decreasing levels ofARA in the subcutaneous adi-
`pose tissue; and decreasing availability of substrates to
`decrease the activity of the endocannabinoid system.
`[0067] The present invention also relates to method of
`using krill oil and/or compositions comprising krill oil to
`modulate biological processes selected from the group con-
`sisting of glucose metabolism, lipid biosynthesis, fatty acid
`metabolism, cholesterol biosynthesis, and the mitochondrial
`respiratory chain.
`[0068] The present invention further includes pharmaceu-
`tical and/or nutraceutical formulations made from krill oil,
`methods of making such formulations, and methods of
`administering them to treat risk factors for metabolic, cardio-
`vascular, and inflammatory disorders.
`
`A. Methods of Using Krill Oi