111111
`
`1111111111111111111111111111111111111111111111111111111111111111111111111111
`US 20040241249Al
`
`(19) United States
`(12) Patent Application Publication
`Sampalis
`
`(10) Pub. No.: US 2004/0241249 Al
`Dec. 2, 2004
`( 43) Pub. Date:
`
`(54) KRILLAND/OR MARINE EXTRACTS FOR
`PREVENTION AND/OR TREATMENT OF
`CARDIOVASCULAR DISEASES ARTHRITIS,
`SKIN CANCER DIABETES,
`PREMENSTRUAL SYNDROME AND
`TRANSDERMAL TRANSPORT
`
`(76)
`
`Inventor: Tina Sampalis, Laval (CA)
`
`Correspondence Address:
`CROWELL & MORING LLP
`INTELLECTUAL PROPERTY GROUP
`P.O. BOX 14300
`WASHINGTON, DC 20044-4300 (US)
`
`(21)
`
`Appl. No.:
`
`10/481,040
`
`(22)
`
`PCT Filed:
`
`Jun. 7, 2002
`
`(86)
`
`PCTNo.:
`
`PCT/CA02/00843
`
`Related U.S. Application Data
`
`(60) Provisional application No. 60/298,383, filed on Jun.
`18, 2001.
`
`Publication Classification
`
`(51)
`Int. Cl? .......................... A61K 35/12; A61K 35/60
`(52) U.S. Cl. ............................................ 424/520; 424/523
`
`(57)
`
`ABSTRACT
`
`The present invention relates to a method of treatment
`and/or prevention of cardiovascular disease, rheumatoid
`arthritis, skin cancer, premenstrual syndrome, diabetes and
`transdermal transport enhancement. The method comprises
`the administration of a therapeutically effective amount of
`krill and/or marine oil to a patient. The present invention
`also relates to a composition for the treatment and/or pre(cid:173)
`vention of these diseases.
`
`AKER EXHIBIT 2017 Page 1
`
`

`

`US 2004/0241249 A1
`
`Dec. 2, 2004
`
`1
`
`KRILLAND/OR MARINE EXTRACTS FOR
`PREVENTION AND/OR TREATMENT OF
`CARDIOVASCULAR DISEASES ARTHRITIS, SKIN
`CANCER DIABETES, PREMENSTRUAL
`SYNDROME AND TRANSDERMAL TRANSPORT
`
`BACKGROUND OF THE INVENTION
`
`[0001] 1. Field of the Invention
`
`[0002] This invention relates to multi-therapeutic extracts
`derived from krill and/or marine, which can prevent and/or
`treat several diseases.
`
`[0003] 2. Description of Prior Art
`
`[0004] Krill is the common name for small, shrimp-like
`crustaceans, however not shrimp, that swarm in dense
`shoals, especially in Antarctic waters. It is one of the most
`important food source for fish, some kind of birds and
`especially for baleen whales as being an important source of
`protein. Krill is also a good source of omega-3 fatty acid,
`which are well known for their health benefits.
`
`[0005]
`It is known in the art to use krill and/or marine
`enzymes for the treatment of a great variety of diseases in
`human and animals such as infections, inflammations, can(cid:173)
`cers, HIV/AIDS, pain, polyps, warts, hemorrhoids, plaque,
`wrinkles, thin hairs, allergic itch, anti-adhesion, eye disease,
`acne, cystic fibrosis and immune disorders
`including
`autoimmune disease and cancer.
`
`[0006]
`It is also known in the art that krill and/or marine
`oil may be used for the treatment of autoimmune murine
`lupus and other autoimmune diseases and can also be used
`for treating cardiovascular diseases.
`
`[0007] However, the krill and/or marine oil used for these
`treatments has only conserved its omega-3 fatty acids as
`active ingredients, which is a very small part of all the active
`ingredients of the krill and/or marine itself. This fact reduces
`the potential of the krill and/or marine oil as a treatment for
`these diseases.
`
`[0008] There is an increasing demand for treatments using
`products derived from a natural source, therefore, it would
`be highly desirable to be provided with a krill and/or marine
`extract having an enhanced potential for prevention and/or
`treatment and/or management of disease.
`
`SUMMARY OF THE INVENTION
`
`[0009]
`In accordance with the present invention there is
`provided a method of prevention, therapy and/or treatment
`of several disease, the method comprising the administration
`of a therapeutically effective amount of krill and/or marine
`oil to a patient.
`
`[0010]
`In a preferred embodiment of the present invention
`the krill and/or marine oil is obtained from a process
`comprising the steps of:
`
`[0011]
`(a) placing krill and/or marine material in a
`ketone solvent, preferably acetone to achieve extrac(cid:173)
`tion of the soluble lipid fraction from the manne
`and/or aquatic animal material;
`
`[0012]
`
`(b) separating the liquid and solid contents;
`
`[0013]
`(c) recovering a first lipid rich fraction from
`the liquid contents by evaporation of the solvent
`present in the liquid contents;
`
`[0014]
`(d) placing the solid contents in an organic
`solvent selected from the group of solvents consist(cid:173)
`ing of alcohol, preferably ethanol, isopropanol or
`t-butanol and esters of acetic acid, preferably ethyl
`acetate to achieve extraction of the remaining soluble
`lipid fraction from the marine and/or aquatic mate(cid:173)
`rial;
`
`[0015]
`
`(e) separating the liquid and solid contents;
`
`[0016]
`(f) recovering a second lipid rich fraction by
`evaporation of the solvent from the liquid contents;
`and
`
`[0017]
`
`(g) recovering the solid contents.
`
`[0018]
`In a preferred embodiment of the present invention,
`the krill and/or marine oil comprises Eicosapentanoic acid,
`Docosahexanoic acid, Phosphatidylcholine, Phosphatidyli(cid:173)
`nositol, Phosphatidylserine, Phosphatidylethanolamine,
`Sphingomyelin, a -tocopherol, all-trans retinol, As tax an thin
`and flavonoid.
`
`[0019]
`In another embodiment of the present invention,
`the krill and/or marine oil comprises Eicosapentanoic acid,
`Docosahexanoic acid, Linolenic acid, Alpha-linolenic acid,
`Linoleic acid, Arachidonic acid, Oleic acid, palmitic acid,
`palmitoleic acid, stearic acid, nervonic acid, Phosphatidyl(cid:173)
`choline, Phosphatidylinositol, Phosphatidylserine, Phos(cid:173)
`phatidylethanolamine, Sphingomyelin, Cholesterol, Triglyc(cid:173)
`erides, Monoglycerides, a-tocopherol, all-trans retinol,
`Astaxanthin, Canthaxanthin, ~-carotene, flavonoid, Zinc,
`Selenium, sodium, potassium and calcium.
`
`[0020]
`In another embodiment of the present invention,
`the krill and/or marine oil comprises Eicosapentanoic acid,
`Docosahexanoic acid, Linolenic acid, Alpha-linolenic acid,
`Linoleic acid, Arachidonic acid, Oleic acid, palmitic acid,
`palmitoleic acid, stearic acid, Phosphatidylcholine, Phos(cid:173)
`phatidy !inositol, Phosphatidy lserine, Phosphatidy lethanola(cid:173)
`mine,
`Sphingomyelin,
`Cholesterol,
`Triglycerides,
`Monoglycerides, a-tocopherol, all-trans retinol, Astaxan(cid:173)
`thin, Canthaxanthin, ~-carotene, Zinc and Selenium.
`
`[0021] The diseases that can be treated and/or prevented
`by the method of the present invention are cardiovascular
`diseases, arthritis, skin cancer, diabetes, premenstrual syn(cid:173)
`drome and transdermal transport enhancement.
`
`[0022]
`In accordance with the present invention there is
`also provided a composition for the treatment and/or pre(cid:173)
`vention and/or therapy of the previously mentioned diseases,
`the composition comprising a therapeutically effective
`amount of krill and/or marine oil in association with a
`pharmaceutically acceptable carrier.
`
`[0023]
`In accordance with the present invention, 1t 1s
`further provided the use of krill and/or marine oil for the
`treatment and/or prevention and/or therapy of the previously
`mentioned diseases.
`
`[0024]
`In accordance with the present invention, it is also
`provided the use of krill and/or marine oil for the manufac(cid:173)
`ture of a medicament for the treatment and/or prevention
`and/or therapy of the previously mentioned diseases.
`
`AKER EXHIBIT 2017 Page 2
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`

`

`US 2004/0241249 Al
`
`Dec. 2, 2004
`
`2
`
`DETAILED DESCRIPTION OF 1HE
`INVENTION
`
`[0025]
`In accordance with the present invention, there is
`provided krill and/or marine extract for prevention and/or
`treatment and/or therapy of several diseases.
`[0026] A multi-therapeutic oil extract free of enzyme is
`derived from krill and/or marine, found in any marine
`environment around the world, for example, the Antarctic
`ocean (euphasia superba), the Pacific ocean (euphasia paci(cid:173)
`fica), the Atlantic ocean, the Indian ocean, in particular
`coastal regions of Mauritius Island and/or Reunion Island of
`Madagascar, Canadian West Coast, Japanese Coast, St(cid:173)
`Lawrence Gulf and Fundy Bay, and this oil extract is a free
`fatty acid lipid fraction.
`[0027] The extraction process can be described as the
`following:
`
`[0028]
`(a) Placing marine and/or aquatic krill and/or
`marine in a ketone solvent, preferably acetone, to
`achieve the extraction of grease from the krill and/or
`marine;
`
`[0029]
`
`(b) Separating the liquid and the solid phases;
`
`[0030]
`(c) Recovering a lipid rich fraction from the
`liquid phase obtained at step (b) by evaporation of
`the solvent present in the liquid phase;
`
`[0031]
`(d) Placing the solid phase in an organic
`solvent, which can be alcohol, preferably ethanol,
`isopropanol or t-butanol, or esters of acetic acid,
`preferably ethyl acetate. This in order to extract the
`remaining soluble lipid fraction from the solid phase;
`
`[0032]
`and
`
`(e) Separating the liquid and the solid phases;
`
`(t) Recovering a lipid rich fraction from the
`[0033]
`liquid phase obtained at step (e) by evaporation of
`the solvent present in the liquid phase.
`
`[0034] The active components of the enzyme-free krill
`and/or marine oil extract are:
`[0035] Lipids
`
`[0036]
`
`i) Omega-3:
`
`[0037]
`
`i. Eicosapentanoic acid: >8 g/100 g
`
`[0038]
`
`ii. Docosahexanoic acid: >2 g/100 g
`
`[0039]
`
`iii. Linolenic acid: >0.10 g/100 g
`
`[0040]
`
`iv. Alpha-linolenic acid: >0.3 g/100 g
`
`[0041]
`In the preferred embodiment of the present inven(cid:173)
`tion, the Omega-3 are found in more than 30 g/100 g.
`
`[0048] Phospholipids
`[0049] Phosphatidylcholine: >4.5 g/100 g
`[0050] Phosphatidylinositol: >107 mg/100 g
`
`[0051] Phosphatidylserine: > 75 mg/100 g
`
`[0052] Phosphatidylethanolamine: >0.5 g/100 g
`
`[0053] Sphingomyelin: >107 mg/100 g
`
`[0054] Neutral Lipids
`
`[0055] Cholesterol: <3 g/100 g
`
`[0056] Triglycerides: <55 g/100 g
`
`[0057] Monoglycerides: >0.5 g/100 g
`
`[0058]
`In another embodiment of the present invention,
`the neutral lipids of the krill and/or marine extract also
`comprises:
`
`[0059] Diglycerides: >0.5 g/100 g
`
`[0060] Antioxydants
`
`[0061] a-tocopherol (vitamin E): >1.0 IU/100 g
`
`[0062] all-trans retinol (vitamin A): >1500 IU/100 g
`
`[0063]
`
`~-carotene: >3000 ,ug/100 ml
`
`[0064] Pigments
`
`[0065] Astaxanthin: >20 mg/100 g
`
`[0066] Canthaxanthin: >2 mg/100 g
`
`[0067] Metals
`
`[0068] Zinc: >0.1 mg/100 g
`
`[0069] Selenium: >0.1 mg/100 g
`
`[0070]
`In another embodiment of the present invention,
`the krill and/or marine extract also comprises:
`
`[0071] Flavonoids: >0.5 mg/100 g
`
`[0072] Sodium: <500 mg/100 g
`
`[0073] Calcium: >0.1 mg/100 g
`
`[0074] Potassium: >50 mg/100 g
`
`[0075] Aluminum: <8.5 mg/100 g
`
`[0076] Protein: >4 g/100 g
`
`[0077] Moisture and volatile matter: <0.8%
`
`[0078] After characterization of the krill and/or marine oil
`extract, it was determined that the extract contains less than
`25 ppm of solvent residue from the extraction process.
`
`[0079] The oil has the following stability indexes:
`
`[0042]
`
`ii) Omega-6: i. Linoleic acid: >0.9 g/100 g
`
`[0080] Peroxide value: <0.1 (mEq/kg)
`
`[0043]
`ii. Arachidonic acid: <0.45 g/100 g, preferably
`<0.6 g/100 g
`
`[0044]
`
`iii) Omega-9: i. Oleic acid: >5 g/100 g
`
`[0081] Oil Stability index: <0.1 after 50 hours at 97.8°
`c.
`[0082] Saponification index: 70-180
`
`[0045]
`
`iv) palmitic acid: >10 g/100 g
`
`[0046] v) palmitoleic acid: 0.08 g/100 g
`
`[0047] vi) stearic acid: >0.5 g/100 g
`
`[0083]
`
`Iodine value:60-130%
`
`[0084] The present invention will be more readily under(cid:173)
`stood by referring to the following examples which are
`given to illustrate the invention rather than to limit its scope.
`
`AKER EXHIBIT 2017 Page 3
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`

`

`US 2004/0241249 Al
`
`Dec. 2, 2004
`
`3
`
`EXAMPLE 1
`
`Cardiovascular Disease Prevention and/or
`Treatment
`
`[0085] Krill and/or marine oil has been shown to decrease
`cholesterol in vivo. It also inhibits platelet adhesion and
`plaque formation and reduces vascular endothelial inflam(cid:173)
`mation in a patient. It can offer hypertension prophylaxis. It
`prevents oxidation of low-density lipoprotein. It may have
`an inhibitory effect on the secretion of VLDL due to
`increased intracellular degradation of apo B-100. It also
`offers a post-myocardial infarction prophylaxis because of
`its ability to decrease CIII apolipoprotein B, to decrease CIII
`non-apolipoprotein B lipoproteins and to increase anti(cid:173)
`thrombin III levels. Krill and/or marine oil is suitable for
`prophylactic usage against cardiovascular disease in human
`where cardiovascular disease relates to coronary artery
`disease, hyperlipidemia, hypertension, ischemic disease
`(relating to angina, myocardial infarction, cerebral ischemia,
`shock without clinical or laboratory evidence of ischemia,
`arrhythmia)
`
`[0086] To evaluate the effects of krill and/or marine oil on
`the course of arteriosclerotic coronary artery disease and
`hyperlipidemia, a study was performed (prospective clinical
`trial, statistical significance p<O.OS) with patients with
`known hyperlipidemia.
`
`[0087] A group of 13 patients took krill and/or marine oil
`concentrate gelules. Both fish oil and krill and/or marine oil
`contained equal amounts of omega-3 fatty acids. Recom(cid:173)
`mended dosage is of 1 to 6 capsules per day, each capsule
`containing 800 mg of oil. In this study, each patient took 6
`capsules per day.
`
`[0088] The patients were tested for LDL, HDL, Triglyc(cid:173)
`erides, vital signs, CBC, SGOT/SGPT, y-GT, ALP, Urea,
`Creatine, Glucose, K+, Na+, Ca2 + and total indirect bilirubin
`cholesterol before treatment and also at 2 months.
`
`[0089] Table 1 is showing the results obtained from the
`previously described tests:
`
`[0091] This shows that an uptake of krill extract has a
`beneficial effect on patient suffering from hyperlipidemia,
`which is known to be the primary causative factor of
`atherosclerosis.
`
`EXAMPLE 2
`
`Arthritis Treatment
`
`[0092] Krill and/or marine oil offers symptomatic relief
`for Arthritis where arthritis relates to adult arthritis, Still's
`disease, polyarticular or pauciarticular juvenile rheumatoid
`arthritis, rheumatoid arthritis, osteoarthritis because it has
`been shown that it provides a clinical improvement in
`decreasing the number of tender joints and of analgesics
`consumed daily by decreasing the production of Interleu(cid:173)
`kin-8 and Interleukin-1 in human patients. Patients with a
`bleeding tendency or severe psychiatric disease were
`excluded from the study.
`
`[0093] To evaluate the effects of krill and/or marine oil
`supplementation on the clinical course of osteoarthritis, a
`study was performed (prospective clinical trial, statistical
`significance p<O.OS) with patients diagnosed with and
`treated for osteoarthritis which is Active class I, II or III and
`having treatment with NSAIDs and/or analgesics for at least
`3 months before enrollment.
`
`[0094] A group of 13 patients took krill and/or marine oil
`concentrate capsules at a daily rate of 6 capsules of 800 mg
`krill oil per capsule. The recommended dosage varies
`between 1 and 4.8 grams of pure krill extract per day.
`Patients were asked to follow a normal healthy diet consist(cid:173)
`ing of 20% fat (less than 10% animal fat), 40% protein and
`40% carbohydrates.
`
`[0095] The inclusion criteria for the study are being aged
`between 50 and 65 years, both genders being admissible,
`having a clinical diagnosis of primary osteoarthritis (mild to
`moderate) 6 to 12 months prior to study enrollment includ(cid:173)
`ing pain and stiffness, radiographic confirmation of illness
`prior to enrollment. It also include evidence of measurable
`
`TABLE 1
`
`Paired Samples Test
`Paired Differences
`
`Parameter
`
`Std.
`Error
`
`95% Confidence
`Interval of the
`Difference
`
`Sig. (2-
`
`tested
`
`Mean
`
`SO.
`
`Mean
`
`Lower Upper
`
`t-value
`
`df
`
`tailed)
`
`Cholesterol
`Triglycerides
`HDL
`LDL
`Chol/HDL
`
`.4954
`.3538
`-.2108
`.2846
`.3600
`
`.55800
`.54543
`.29859
`.47333
`.53446
`
`.15476
`.15127
`.08281
`.13128
`.14823
`
`.1582
`.0242
`-.3912
`-.0014
`.0370
`
`.8326
`.6834
`-.0303
`.5706
`.6830
`
`3.201
`2.339
`-2.545
`2.168
`2.429
`
`12
`12
`12
`12
`12
`
`.008
`.037
`.026
`.051
`.032
`
`[0090] From the above, it was shown that a daily uptake
`of 1 to 4.8 g of krill extract was providing to the patients a
`cholesterol decrease in the range of 15%, a triglycerides
`decrease in the range of 15%, a HDL increase in the range
`of 8%, a LDL decrease in the range of 13% and a Choles(cid:173)
`terol/HDL ratio decrease of 14%.
`
`symptoms of OA for at least 3 months prior to study
`enrollment requiring the use of acetaminophen, anti-inflam(cid:173)
`matory agents or opioid analgesics. Patients were asked to
`stop the use of all "pain-killers" the week prior to initiation
`of the trial for wash-out purposes.
`[0096] The Exclusion criteria were a severe osteoarthritis,
`unavoidable sustained use of NSAID's, aspirin or other
`
`AKER EXHIBIT 2017 Page 4
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`

`US 2004/0241249 Al
`
`Dec. 2, 2004
`
`4
`
`medicines for anti-inflammatory use, use of topical analge(cid:173)
`sics within 4 weeks of randomization visit, steroid injection
`into either knee within past 3 months, initiation of physical
`therapy or muscle conditioning within 3 months, seafood
`allergies, use of anticoagulants or salicylates, alcohol con(cid:173)
`sumption exceeding 3 mixed drinks per day, concurrent
`medical/arthritic disease that could confound or interfere
`with the evaluation of pain, prior surgery (including arthros(cid:173)
`copy) of either knee, a known "secondary" cause of osteoar(cid:173)
`thritis.
`
`[0097] Evaluation was based on daily dose of NSAIDs
`and/or analgesics and/or SAARDs, number of painful joints,
`number or swollen joints, duration of morning stiffness,
`visual analog scale (0-100) WOMACscale and SF36. Pre(cid:173)
`liminary results have been obtained after 2 months. The
`number of NSAIDs and/or analgesics and/or SAARDs
`required for daily functioning has been recorded at initiation
`and at 2 months after initiation.
`
`[0098] Results shown at Table 2 demonstrate the effect of
`an uptake of krill extract on the relief of arthritis.
`
`TABLE 2
`
`Frequency
`
`%
`
`Valid%
`
`Cumulative %
`
`No change
`Pain relief
`
`Total
`
`3
`10
`
`13
`
`23.1
`76.9
`
`23.1
`76.9
`
`23.1
`100.0
`
`100.0
`
`100.0
`
`[0099] This shows that ten out of 13 (76.9%) people
`reported a significant pain relief and improvement of flex(cid:173)
`ibility of large joints (lower back, knees, shoulders)
`
`EXAMPLE 3
`
`Skin Cancer Prophylaxis
`
`[0100] Krill and/or marine oil has been shown to be a skin
`cancer prophylactic because of its retinol anti-carcinogenic
`effect, Astaxanthin anti -carcinogenic effect and its phopho(cid:173)
`lipid anti-carcinogenic effect.
`
`[0101] To evaluate the photoprotective potential of krill
`and/or marine oil against UVB-induced skin cancer, a study
`was performed on nude mice, preferably on C57BL6 Nude
`Congenic Mice-B6NU-T (heterozygotes) because of their
`proven susceptibility to skin cancer.
`
`[0102] Groups were formed as follows: 48 fish oil: 16 with
`oral supplementation (po) 16 with local application, 16 with
`po and local application; 48 krill and/or marine oil: 16 with
`po, 16 with local application, 16 with po and local applica(cid:173)
`tion. In order to establish efficacy of krill and/or marine oil
`for the prevention of skin cancer, the test was conducted as
`a randomized blind controlled trial (statistical significance
`p<O.OS). Half of the mice have been treated orally or
`topically or both with oil containing 100% by weight krill
`and/or marine oil and the other half have been treated the
`same way with fish oil.
`
`[0103] Nutrition was fat-free chow for the first week and
`was modified accordingly with the assigned group as
`described below for the following 2-20 weeks in the quantity
`of 1 ml of oil per day.
`
`[0104] The mice were divided in six groups as follows:
`
`[0105] Group A: fat-free chow with supplementation of
`fish oil (20% of total calories)
`
`[0106] Group B: fat-free chow (100% of calories)+local
`application of fish oil 2 times per day
`
`[0107] Group C: fat free chow with supplementation of
`fish oil (20% of total calories)+local application of soy
`oil 2 times per day
`
`[0108] Group D: fat-free chow with supplementation of
`krill and/or marine oil (20% of total calories)
`
`[0109] Group E: fat free chow (100% of calories)+local
`application of krill and/or marine oil 2 times per day
`
`[0110] Group F: fat-free chow with supplementation or
`krill and/or marine oil (20% of total calories)+local
`application of krill and/or marine oil 2 times per day
`
`[0111] The mice had been submitted to UVB radiation
`using a fluorescent test lamp, emission spectrum 270400 nm
`during weeks 2-20. The essay were performed during 30
`minutes of UVB exposure per day and the test lamp was at
`a distance of 30 em from the mice. At the end of the 20
`weeks, or when malignant tumors had formed, mice were
`anesthetized with ether and sacrificed. Skin was examined
`blind by pathologists for signs of carcinogenesis.
`
`[0112] The following tables (Tables 3-8) are showing the
`results obtained about the incidence of cancer when ultra(cid:173)
`violet radiations are administered to mice's skin during 5
`weeks.
`
`TABLE 3
`
`Krill extract Oral uptake
`
`Frequency Percent Valid Percent
`
`Percent
`
`Cumulative
`
`Valid
`
`Benign
`
`Cancer
`
`14
`
`2
`
`87.5
`
`12.5
`
`87.5
`
`12.5
`
`87.5
`
`100.0
`
`Total
`
`16
`
`100.0
`
`100.0
`
`[0113]
`
`TABLE 4
`
`Control Oral uptake
`
`Frequency Percent Valid Percent
`
`Cumulative
`Percent
`
`Valid
`
`Benign
`Cancer
`
`Total
`
`14
`2
`
`16
`
`87.5
`12.5
`
`87.5
`12.5
`
`87.5
`100.0
`
`100.0
`
`100.0
`
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`US 2004/0241249 Al
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`Dec. 2, 2004
`
`5
`
`[0114]
`
`TABLE 5
`
`Krill extract toQical UQtake
`
`Frequency Percent Valid Percent
`
`Cumulative
`Percent
`
`Valid
`
`BENIGN
`
`16
`
`100.0
`
`100.0
`
`100.0
`
`[0115]
`
`TABLE 6
`
`Control toQical UQtake
`
`Frequency Percent Valid Percent
`
`Cumulative
`Percent
`
`Valid
`
`BENIGN
`Cancer
`
`Total
`
`5
`11
`
`16
`
`31.3
`68.8
`
`31.3
`68.8
`
`31.3
`100.0
`
`100.0
`
`100.0
`
`[0116]
`
`TABLE 7
`
`Krill extract toQical and oral UQtake
`
`Frequency Percent Valid Percent
`
`Cumulative
`Percent
`
`Valid
`
`BENIGN
`
`16
`
`100.0
`
`100.0
`
`100.0
`
`[0117]
`
`TABLE 8
`
`Control toQical and oral UQtake
`
`Frequency Percent Valid Percent
`
`Cumulative
`Percent
`
`Valid
`
`BENIGN
`Cancer
`
`10
`
`62.5
`37.5
`
`62.5
`37.5
`
`62.5
`100.0
`
`Total
`
`16
`
`100.0
`
`100.0
`
`mal absorption of krill and/or marine alone or as a substrate,
`a study was performed as a randomized blind controlled trial
`on C57BL6 nude Congenic Mice-B6NU-T (heterozygotes).
`
`[0121] The results appearing in tables 5 and 6 are showing
`that topical treatment with krill oil faciliate the absorption of
`retinol and other antioxydants through the dermis which in
`turn result in significant photoprotective potential which in
`turn results in 100% protection from UVB induced skin
`cancer. In contrast, fish oil application with all-trans retinol
`resulted in 68.8% incidence of cancer.
`
`EXAMPLE 5
`
`Transdermal Transport for Dermatological Topical
`Cosmetic Applications
`
`[0122] Krill and/or marine oil can be used to enhance
`transdermal transportation as a substrate for dermatological
`topical cosmetic applications where cosmetic applications
`relate to skin hydration, anti -wrinkle, keratolytics, peeling
`and mask via creams, ointments, gels, lotions or oils.
`
`[0123] To evaluate the effects of Krill and/or marine oil in
`aging and facial wrinkles, a study was conducted as a
`prospective clinical trial on patients concerned about facial
`dryness and wrinkles. Those patients had no prognosis
`severely limited by other dermatological or non-dermato(cid:173)
`logical condition, bleeding tendency or severe psychiatric
`disease.
`
`[0124] 13 Healthy Caucasian women with facial dryness
`or wrinkles have been included in this study. Women have
`been asked to take 6 capsules a day, each capsule containing
`800 mg of krill extract. The recommended daily dosage is of
`about 1 to 4.8 g of krill extract.
`
`[0125] Table 9 shows results obtained on skin hydration
`following the method previously described.
`
`TABLE 9
`
`Changes in skin hydration
`
`Frequency
`
`%
`
`Valid%
`
`Cumulative %
`
`No change
`Hydration
`
`Total
`
`4
`9
`
`13
`
`30.8
`69.2
`
`30.8
`69.2
`
`30.8
`100.0
`
`100.0
`
`100.0
`
`[0118] The results obtained shows that both oral and
`topical use of krill oil is effective for the protection of the
`skin against the harmful effects fo UVB radiation induced
`skin cancer.
`
`EXAMPLE 4
`
`Transdermal Transport in Therapeutic Applications
`
`[0119] Krill and/or marine oil enhances transdermal trans(cid:173)
`portation as a substrate for dermatological topical therapeu(cid:173)
`tic applications. It may be used in dermatological treatments
`via creams, ointments, gels, lotions and oils. It may also be
`used in various therapeutic applications such as relating to
`anesthesic, corticosteroids, anti-inflammatory, antibiotic and
`ketolytic functions.
`
`[0120] To evaluate the efficacy of krill and/or marine oil as
`a substrate for topical treatments and the speed of transder-
`
`[0126] The results of the pilot study after 2 months indi(cid:173)
`cate that nine out of 13 (69.2%) people reported a significant
`improvement of the hydration, texture and elasticity of the
`skin (face, hands and arms) in human patients.
`
`[0127] Moreover, these results are also indicative that krill
`extract is useful for anti-wrinkle treatment. The mechanism
`of all-trans retinol, which is included in the krill oil, as an
`anti-wrinkle works as follows:
`
`[0128] Regeneration and distinctive anti-inflamma(cid:173)
`tory effects
`
`[0129]
`
`Improve blood irrigation
`
`[0130]
`the epidermis regeneration by
`Increases
`increasing the rate of cell division and turnover
`
`[0131] Accelerates the differentiation of keratin
`
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`

`US 2004/0241249 Al
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`
`[0132] Regenerates the collagen
`
`[0133] Allows cells in the top layer of the skin, which
`are always being replaces, to mature more normally
`than untreated sun-damaged cells
`
`[0134] Reduces the activation of enzymes that break
`down the proteins collagen and elastin that provide
`structural support for the skin.
`
`[0135] The results obtained with krill extract administered
`on a patent's skin show that the krill extract is having an
`anti-wrinkle effect by increasing the hydration and the
`mechanism above described.
`
`EXAMPLE 6
`
`Premenstrual Syndrome
`
`[0136] Table 10 shows results obtained from the use of
`krill oil to reduce the pain and mood changes associated with
`premenstrual syndrome in women. Krill oil extract was
`administered to 7 women during 2 months. The women were
`taking 6 capsules of krill extract per day, each capsule
`containing 800 mg of krill oil. A recommended daily intake
`of krill oil is of about 1 to 4.8 grams. All participants were
`advised to continue with their usual nutrition habits and to
`refrain from initiating any restrictions in their diet. No
`serious side effects were reported.
`
`[0137] All women enrolled reported noticeable emotional
`and/or physical discomfort 7 to 10 days prior to menstrua(cid:173)
`tion. A self-assessment visual analogue scale validated for
`the assessment of the premenstrual syndrome, ranging from
`0 (no symptoms) to 10 (unbearable) was used as a primary
`outcome in order to evaluate the effect of krill extract on
`premenstrual discomfort.
`
`[0138] Data analysis has been reported on 60% of the
`women participating in the study who have completed a two
`months regimen. The majority of the women (73.3%)
`showed a clinically significant reduction in both emotional
`and physical distress prior to menstruation (see Table 10).
`
`TABLE 10
`
`Frequency distribution of the effect of krill extract on
`premenstrual syndrome symptomatology
`
`PMS symptoms
`
`Frequency %
`
`Valid %
`
`Cumulative %
`
`No change
`Positive
`
`26.7
`73.3
`
`26.7
`73.3
`
`26.7
`100.0
`
`Total
`
`100.0
`
`100.0
`
`EXAMPLE 7
`
`Diabetes
`
`[0139] 8 human patients were taking krill extract at the
`dosage of 6 capsules a day, each capsule containing 800 mg
`of krill extract, during 2 months. A recommended daily
`intake of krill oil is of about 1 to 4.8 grams. The Table 11 is
`showing the variation in the glucose tested for the patients
`after 2 months.
`
`TABLE 11
`
`Variation in glucose in patients
`Paired Differences
`
`Para-
`meter
`tested
`
`Mean
`
`SO.
`
`95%
`Confidence
`Interval
`of the
`Difference
`
`Std.
`Error
`Mean
`
`t-
`value df
`
`Sig.
`(2-
`tailed)
`
`Glucose
`
`.5778
`
`.60369
`
`.20123
`
`.1137-1.0418
`
`2.871
`
`8
`
`.021
`
`[0140] A blood glucose decrease of 20% was obtained for
`the patients taking krill extract, which shows that an uptake
`of krill extract is controlling blood glucose content and
`therefore controlling diabetes in human patients.
`
`[0141] While the invention has been described in connec(cid:173)
`tion with specific embodiments thereof, it will be understood
`that it is capable of further modifications and this application
`is intended to cover any variations, uses, or adaptations of
`the invention following, in general, the principles of the
`invention and including such departures from the present
`disclosure as come within known or customary practice
`within the art to which the invention pertains and as may be
`applied to the essential features hereinbefore set forth, and
`as follows in the scope of the appended claims.
`
`1-83. (Cancelled)
`84. A composition comprising an amount of krill oil
`effective for decreasing cholesterol, inhibiting platelet adhe(cid:173)
`sion, inhibiting artery plaque formation, preventing hyper(cid:173)
`tension, controlling arthritis symptoms, preventing skin can(cid:173)
`cer, enhancing transdermal transportation of dermatological
`topical therapeutic applications, enhancing transdermal
`transportation of dermatological cosmetic applications,
`reducing symptoms of premenstrual syndrome, or control(cid:173)
`ling blood glucose level in a patient, and a pharmaceutically
`acceptable carrier, wherein said krill oil is obtained from a
`process comprising the steps of:
`a) placing marine or aquatic krill in a ketone solvent, to
`obtain a first extraction of a soluble lipid fraction from
`said marine or aquatic krill;
`b) separating the extraction into a first liquid and a first
`solid contents;
`c) evaporating the ketone solvent present in the first liquid
`contents to recover a first lipid rich fraction;
`d) placing said first solid contents in an organic solvent
`selected from the group of consisting of alcohol and
`esters of acetic acid, to achieve a second extraction of
`soluble lipid fraction;
`e) separating the second extraction into a second liquid
`and a second solid contents;
`f) evaporating the organic solvent from the second liquid
`contents to recover a second lipid rich fraction; and
`g) recovering the solid contents.
`85. A composition according to claim 84, wherein the
`ketone solvent is acetone.
`86. A composition according to claim 84, wherein the
`organic solvent is ethanol, isopropanol, t-butanol, or ethyl
`acetate
`87. A composition according to claim 84, wherein the
`composition comprises Eicosapentanoic acid, Docosahex-
`
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`US 2004/0241249 Al
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`
`7
`
`anoic acid, Phosphatidylcholine, Phosphatidylinositol,
`Phosphatidylserine, Phosphatidylethanolamine, Sphingo(cid:173)
`myelin, a-tocopherol, Astaxanthin, and flavonoid.
`88. A composition of claim 87, further comprising at least
`one of the group consisting of Linoleic acid, Alpha-linoleic
`acid, arachidonic acid, oleic acid, palmitic acid, palmitoleic
`acid, stearic acid, cholesterol, triglycerides, monoglycerides,
`all-trans retinol, canthaxanthin, ~-carotene, zinc, selenium,
`nervonic acid, sodium, potassium and calcium.
`89. A composition of claim 84, wherein the composition
`comprising an effective amount for decreasing cholesterol in
`a patient.
`90. A composition of claim 84, wherein the composition
`comprising an effective amount for inhibiting platelet adhe(cid:173)
`sion or artery plaque formation in a patient.
`91. A composition of claim 84, wherein the composition
`comprising an effective amount for preventing hypertension
`in a patient.
`92. A composition of claim 84, wherein the composition
`comprising an effective amount for controlling arthritis
`symptoms in a patient.
`93. A composition of claim 92, wherein the arthritis is
`rheumatoid arthritis or osteoarthritis.
`94. A composition of claim 84, wherein the composition
`comprising an effective amount for preventing skin cancer in
`a patient.
`95. A composition of claim 84, wherein the composition
`comprising an effectiv