(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2004/0241249 A1
`Sampalis
`(43) Pub. Date:
`Dec. 2, 2004
`
`US 2004O241.249A1
`
`(54) KRILL AND/OR MARINE EXTRACTS FOR
`PREVENTION AND/OR TREATMENT OF
`CARDIOVASCULAR DISEASES ARTHRITIS,
`SKIN CANCER DIABETES,
`PREMENSTRUAL SYNDROME AND
`TRANSIDERMAL TRANSPORT
`
`Related U.S. Application Data
`(60) Provisional application No. 60/298,383, filed on Jun.
`18, 2001.
`
`Publication Classification
`
`(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) PCT No.:
`
`PCT/CA02/00843
`
`(51) Int. C.7 - - - - - - - - - - - - - - - - - - - - - - - - - - A61 K 35/12; A61 K 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
`vention of these diseases.
`
`RIMFROST EXHIBIT 1168 Page 0000
`
`

`

`US 2004/0241249 A1
`
`Dec. 2, 2004
`
`KRILL AND/OR MARINE EXTRACTS FOR
`PREVENTION AND/OR TREATMENT OF
`CARDIOVASCULAR DISEASES ARTHRITIS, SKIN
`CANCER DIABETES, PREMENSTRUAL
`SYNDROME AND TRANSIDERMAL 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
`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
`tion of the soluble lipid fraction from the marine
`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
`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
`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
`nositol, Phosphatidylserine, Phosphatidylethanolamine,
`Sphingomyelin, a-tocopherol, all-trans retinol, AStaxanthin
`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
`choline, Phosphatidylinositol, Phosphatidylserine, Phos
`phatidylethanolamine, Sphingomyelin, Cholesterol, Triglyc
`erides, Monoglycerides, a-tocopherol, all-trans retinol,
`AStaxanthin, Canthaxanthin, B-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
`phatidylinositol, Phosphatidylserine, Phosphatidylethanola
`mine,
`Sphingomyelin,
`Cholesterol,
`Triglycerides,
`Monoglycerides, a-tocopherol, all-trans retinol, AStaxan
`thin, Canthaxanthin, B-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
`drome and transdermal transport enhancement.
`0022. In accordance with the present invention there is
`also provided a composition for the treatment and/or pre
`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, it is
`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
`ture of a medicament for the treatment and/or prevention
`and/or therapy of the previously mentioned diseases.
`RIMFROST EXHIBIT 1168 Page 0001
`
`

`

`US 2004/0241249 A1
`
`Dec. 2, 2004
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`In accordance with the present invention, there is
`0.025
`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
`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
`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 (e) Separating the liquid and the solid phases;
`and
`0033 (f) Recovering a lipid rich fraction from the
`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
`i) Omega-3:
`0036)
`i. Eicosapentanoic acid: >8 g/100 g
`0037)
`ii. Docosahexanoic acid: >2 g/100 g
`0038
`iii. Linolenic acid: >0.10 g/100 g
`0039)
`iv. Alpha-linolenic acid: >0.3 g/100 g
`0040
`0041. In the preferred embodiment of the present inven
`tion, the Omega-3 are found in more than 30 g/100 g.
`0042 ii) Omega-6: i. Linoleic acid: >0.9 g/100 g
`0043 ii. Arachidonic acid: <0.45 g/100 g, preferably
`<0.6 g/100 g
`iii) Omega-9: i. Oleic acid: >5 g/100 g
`0044)
`iv) palmitic acid: >10 g/100 g
`0045
`0046) v) palmitoleic acid: 0.08 g/100 g
`0047 vi) stearic acid: >0.5 g/100 g
`
`0048 Phospholipids
`0049) Phosphatidylcholine: >4.5 g/100 g
`0050 Phosphatidylinositol: >107 mg/100 g
`0051 Phosphatidylserine: >75 mg/100 g
`0.052
`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 C-tocopherol (vitamin E): >1.0 IU/100 g
`0062) all-trans retinol (vitamin A); >1500 IU/100 g
`0063
`B-carotene: >3000 lug/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
`0.076
`Protein: >4 g/100 g
`0.077
`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.
`007.9 The oil has the following stability indexes:
`0080 Peroxide value: <0.1 (mEq/kg)
`0081) Oil Stability index: <0.1 after 50 hours at 97.8
`C.
`0082 Saponification index: 70-180
`0083 Iodine value:60-130%
`0084. The present invention will be more readily under
`stood by referring to the following examples which are
`given to illustrate the invention rather than to limit its Scope.
`RIMFROST EXHIBIT 1168 Page 0002
`
`

`

`US 2004/0241249 A1
`
`Dec. 2, 2004
`
`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
`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
`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)
`0.086 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<0.05) 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
`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
`erides, vital signs, CBC, SGOT/SGPT, Y-GT, ALP, Urea,
`Creatine, Glucose, K", Na", Cat" 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, Stills
`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
`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<0.05) 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
`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
`ing pain and Stiffness, radiographic confirmation of illness
`prior to enrollment. It also include evidence of measurable
`
`TABLE 1.
`
`Paired Samples Test
`Paired Differences
`
`Std.
`Error
`
`95% Confidence
`Interval of the
`Difference
`
`Parameter
`
`tested
`
`Mean
`
`SD.
`
`Mean Lower Upper t-value
`
`df
`
`Cholesterol
`Triglycerides
`HDL
`LDL
`Cholf HDL
`
`3.2O1
`8326
`1582
`15476
`4954 .558OO
`2.339
`6834
`O242
`15127
`3538 .54543
`-.2108 .29859 .08.281 -3912 -.0303 -2.545
`2846 .47333
`13128 - OO14
`57O6
`2.168
`3600 .53446 .14823
`O370
`683O
`2.429
`
`12
`12
`12
`12
`12
`
`Sig. (2-
`
`tailed)
`
`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
`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
`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
`RIMFROST EXHIBIT 1168 Page 0003
`
`

`

`US 2004/0241249 A1
`
`Dec. 2, 2004
`
`medicines for anti-inflammatory use, use of topical analge
`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
`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
`copy) of either knee, a known “secondary cause of Osteoar
`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
`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.
`0.098
`Results shown at Table 2 demonstrate the effect of
`an uptake of krill extract on the relief of arthritis.
`
`TABLE 2
`
`Frequency
`
`%
`
`Walid 2%
`
`Cumulative 26
`
`No change
`Pain relief
`
`Total
`
`3
`1O
`
`13
`
`23.1
`76.9
`
`23.1
`76.9
`
`23.1
`1OO.O
`
`1OO.O
`
`1OO.O
`
`0099] This shows that ten out of 13 (76.9%) people
`reported a significant pain relief and improvement of flex
`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
`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
`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<0.05). 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
`0.108 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 cm 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
`Violet radiations are administered to mice’s skin during 5
`weeks.
`
`TABLE 3
`
`Krill extract Oral uptake
`
`Frequency Percent Valid Percent
`
`Cumulative
`Percent
`
`Walid
`
`Benign
`Cancer
`
`14
`2
`
`87.5
`12.5
`
`87.5
`12.5
`
`87.5
`1OO.O
`
`Total
`
`16
`
`1OO.O
`
`1OO.O
`
`0113)
`
`TABLE 4
`
`Control Oral uptake
`
`Frequency Percent Valid Percent
`
`Cumulative
`Percent
`
`Walid
`
`Benign
`Cancer
`
`Total
`
`14
`2
`
`16
`
`87.5
`12.5
`
`87.5
`12.5
`
`87.5
`1OO.O
`
`1OO.O
`
`1OO.O
`
`RIMFROST EXHIBIT 1168 Page 0004
`
`

`

`US 2004/0241249 A1
`
`Dec. 2, 2004
`
`0114
`
`TABLE 5
`
`Krill extract topical uptake
`
`Frequency Percent
`
`Valid Percent
`
`Cumulative
`Percent
`
`Walid
`
`BENIGN
`
`16
`
`1OO.O
`
`1OO.O
`
`1OO.O
`
`0115)
`
`TABLE 6
`
`Qontrol topical uptake
`
`Frequency Percent
`
`Valid Percent
`
`Cumulative
`Percent
`
`Walid
`
`BENIGN
`Cancer
`
`Total
`
`5
`11
`
`16
`
`31.3
`68.8
`
`31.3
`68.8
`
`31.3
`1OO.O
`
`1OO.O
`
`1OO.O
`
`0.116)
`
`TABLE 7
`
`Krill extract topical and oral uptake
`
`Frequency Percent
`
`Valid Percent
`
`Cumulative
`Percent
`
`Walid
`
`BENIGN
`
`16
`
`1OO.O
`
`1OO.O
`
`1OO.O
`
`0117)
`
`TABLE 8
`
`Control topical and Oral uptake
`
`Frequency Percent
`
`Valid Percent
`
`Cumulative
`Percent
`
`Walid
`
`BENIGN
`Cancer
`
`Total
`
`1O
`6
`
`16
`
`62.5
`37.5
`
`62.5
`37.5
`
`62.5
`1OO.O
`
`1OO.O
`
`1OO.O
`
`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
`portation as a Substrate for dermatological topical therapeu
`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
`
`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
`Krill and/or marine oil can be used to enhance
`0.122
`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
`logical condition, bleeding tendency or Severe psychiatric
`disease.
`13 Healthy Caucasian women with facial dryness
`0.124
`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.
`0.125
`Table 9 shows results obtained on skin hydration
`following the method previously described.
`
`TABLE 9
`
`Changes in skin hydration
`
`Frequency
`
`%
`
`Walid 2%
`
`Cumulative 26
`
`No change
`Hydration
`
`Total
`
`4
`9
`
`13
`
`30.8
`69.2
`
`30.8
`69.2
`
`30.8
`1OO.O
`
`1OO.O
`
`1OO.O
`
`0.126 The results of the pilot study after 2 months indi
`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
`tory effects
`Improve blood irrigation
`0129
`0.130
`Increases the epidermis regeneration by
`increasing the rate of cell division and turnover
`0131 Accelerates the differentiation of keratin
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`
`0132) Regenerates the collagen
`0.133 Allows cells in the top layer of the skin, which
`are always being replaces, to mature more normally
`than untreated Sun-damaged cells
`0.134
`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
`Table 10 shows results obtained from the use of
`0.136
`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.
`0.137
`All women enrolled reported noticeable emotional
`and/or physical discomfort 7 to 10 days prior to menstrua
`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.
`0.138. 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 96
`
`Valid 7%
`
`Cumulative 26
`
`No change
`Positive
`
`26.7
`73.3
`
`26.7
`73.3
`
`26.7
`1OO.O
`
`Total
`
`1OO.O
`
`1OO.O
`
`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 SD.
`
`95%
`Confidence
`Interval
`of the
`Difference
`
`Std.
`Error
`Mean
`
`Sig.
`(2-
`t-
`value df tailed)
`
`Glucose .5778 .60369 .20123
`
`1137-1.0418 2.871 8
`
`O21
`
`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
`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
`Sion, inhibiting artery plaque formation, preventing hyper
`tension, controlling arthritis Symptoms, preventing skin can
`cer, enhancing transdermal transportation of dermatological
`topical therapeutic applications, enhancing transdermal
`transportation of dermatological cosmetic applications,
`reducing Symptoms of premenstrual Syndrome, or control
`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
`RIMFROST EXHIBIT 1168 Page 0006
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`US 2004/0241249 A1
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`Dec. 2, 2004
`
`anoic acid, Phosphatidylcholine, Phosphatidylinositol,
`Phosphatidylserine, Phosphatidylethanolamine, Sphingo
`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, 3-caroteine, 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
`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 effective amount for enhancing transdermal
`transportation of dermatological topical therapeutic applica
`tions, or enhancing transdermal transportation of dermato
`logical cosmetic applications decreasing cholesterol in a
`patient.
`96. A composition of claim 84, wherein the composition
`comprising an effective amount for reducing Symptoms of
`premenstrual Syndrome in a patient.
`97. The composition of claim 84, wherein the composi
`tion comprising an effective amount for controlling blood
`glucose level in a patient.
`98. A method for decreasing cholesterol, inhibiting plate
`let adhesion, inhibiting artery plaque formation, preventing
`
`hypertension, controlling arthritis Symptoms, preventing
`skin cancer, enhancing transdermal transportation of derma
`tological topical therapeutic applications, enhancing trans
`dermal transportation of dermatological cosm