US20040241249A1 - Krill and/or marine extracts for prevention and/or treatment of cardiovascular diseases arthritis, skin cancer diabetes, premenstrual syndrome and transdermal transport - Google Patents

Krill and/or marine extracts for prevention and/or treatment of cardiovascular diseases arthritis, skin cancer diabetes, premenstrual syndrome and transdermal transport Download PDF

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US20040241249A1
US20040241249A1 US10/481,040 US48104004A US2004241249A1 US 20040241249 A1 US20040241249 A1 US 20040241249A1 US 48104004 A US48104004 A US 48104004A US 2004241249 A1 US2004241249 A1 US 2004241249A1
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composition
patient
krill
acid
effective amount
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Tina Sampalis
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Aker Biomarine Antarctic AS
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Priority to US10/481,040 priority Critical patent/US20040241249A1/en
Assigned to NEPTUNE TECHNOLOGIES & BIORESSOURCES INC. reassignment NEPTUNE TECHNOLOGIES & BIORESSOURCES INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: SAMPALIS, TINA
Publication of US20040241249A1 publication Critical patent/US20040241249A1/en
Priority to US11/640,235 priority patent/US8057825B2/en
Priority to US13/216,694 priority patent/US20110305771A1/en
Priority to US14/152,603 priority patent/US20140199414A1/en
Priority to US15/267,988 priority patent/US20170224742A1/en
Assigned to AKER BIOMARINE ANTARCTIC AS reassignment AKER BIOMARINE ANTARCTIC AS ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: Neptune Technologies & Bioressources, Inc.
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    • A61K35/56—Materials from animals other than mammals
    • A61K35/612—Crustaceans, e.g. crabs, lobsters, shrimps, krill or crayfish; Barnacles
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Definitions

  • This invention relates to multi-therapeutic extracts derived from krill and/or marine, which can prevent and/or treat several diseases.
  • 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.
  • 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.
  • 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.
  • a method of prevention, therapy and/or treatment of several disease comprising the administration of a therapeutically effective amount of krill and/or marine oil to a patient.
  • the krill and/or marine oil is obtained from a process comprising the steps of:
  • 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, Phosphatidylcholine, Phosphatidylinositol, Phosphatidylserine, Phosphatidylethanolamine, Sphingomyelin, Cholesterol, Triglycerides, Monoglycerides, a-tocopherol, all-trans retinol, Astaxanthin, Canthaxanthin, ⁇ -carotene, flavonoid, Zinc, Selenium, sodium, potassium and calcium.
  • 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, Phosphatidylinositol, Phosphatidylserine, Phosphatidylethanolamine, Sphingomyelin, Cholesterol, Triglycerides, Monoglycerides, a-tocopherol, all-trans retinol, Astaxanthin, Canthaxanthin, ⁇ -carotene, Zinc and Selenium.
  • compositions for the treatment and/or prevention and/or therapy of the previously mentioned diseases comprising a therapeutically effective amount of krill and/or marine oil in association with a pharmaceutically acceptable carrier.
  • 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 pacifica), 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.
  • the active components of the enzyme-free krill and/or marine oil extract are:
  • the neutral lipids of the krill and/or marine extract also comprises:
  • ⁇ -tocopherol (vitamin E) >1.0 IU/100 g
  • ⁇ -carotene >3000 ⁇ g/100 ml
  • Astaxanthin >20 mg/100 g
  • Canthaxanthin >2 mg/100 g
  • the krill and/or marine extract also comprises:
  • Flavonoids >0.5 mg/100 g
  • Oil Stability index ⁇ 0.1 after 50 hours at 97.8° C.
  • Table 1 is showing the results obtained from the previously described tests: TABLE 1 Paired Samples Test Paired Differences 95% Confidence Std. Interval of the Parameter Error Difference Sig. (2- tested Mean SD. Mean Lower Upper t-value df tailed) Cholesterol .4954 .55800 .15476 .1582 .8326 3.201 12 .008 Triglycerides .3538 .54543 .15127 .0242 .6834 2.339 12 .037 HDL ⁇ .2108 .29859 .08281 ⁇ .3912 ⁇ .0303 ⁇ 2.545 12 .026 LDL .2846 .47333 .13128 ⁇ .0014 .5706 2.168 12 .051 Chol/HDL .3600 .53446 .14823 .0370 .6830 2.429 12 .032
  • 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 including pain and stiffness, radiographic confirmation of illness prior to enrollment. It also include evidence of measurable symptoms of OA for at least 3 months prior to study enrollment requiring the use of acetaminophen, anti-inflammatory 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.
  • the Exclusion criteria were a severe osteoarthritis, unavoidable sustained use of NSAID's, aspirin or other medicines for anti-inflammatory use, use of topical analgesics 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 consumption exceeding 3 mixed drinks per day, concurrent medical/arthritic disease that could confound or interfere with the evaluation of pain, prior surgery (including arthroscopy) of either knee, a known “secondary” cause of osteoarthritis.
  • 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. Preliminary 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.
  • 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 3 23.1 23.1 23.1 Pain relief 10 76.9 76.9 100.0 Total 13 100.0 100.0
  • 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.
  • mice were divided in six groups as follows:
  • Group E fat free chow (100% of calories)+local application of krill and/or marine oil 2 times per day
  • 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
  • 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.
  • mice were anesthetized with ether and sacrificed. Skin was examined blind by pathologists for signs of carcinogenesis.
  • 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.
  • 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.

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 prevention of these diseases.

Description

    BACKGROUND OF THE INVENTION
  • 1. Field of the Invention [0001]
  • This invention relates to multi-therapeutic extracts derived from krill and/or marine, which can prevent and/or treat several diseases. [0002]
  • 2. Description of Prior Art [0003]
  • 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. [0004]
  • 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, cancers, 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. [0005]
  • 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. [0006]
  • 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. [0007]
  • 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. [0008]
    • SUMMARY OF THE INVENTION
  • 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. [0009]
  • In a preferred embodiment of the present invention the krill and/or marine oil is obtained from a process comprising the steps of: [0010]
  • (a) placing krill and/or marine material in a ketone solvent, preferably acetone to achieve extraction of the soluble lipid fraction from the marine and/or aquatic animal material; [0011]
  • (b) separating the liquid and solid contents; [0012]
  • (c) recovering a first lipid rich fraction from the liquid contents by evaporation of the solvent present in the liquid contents; [0013]
  • (d) placing the solid contents in an organic solvent selected from the group of solvents consisting 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 material; [0014]
  • (e) separating the liquid and solid contents; [0015]
  • (f) recovering a second lipid rich fraction by evaporation of the solvent from the liquid contents; and [0016]
  • (g) recovering the solid contents. [0017]
  • In a preferred embodiment of the present invention, the krill and/or marine oil comprises Eicosapentanoic acid, Docosahexanoic acid, Phosphatidylcholine, Phosphatidylinositol, Phosphatidylserine, Phosphatidylethanolamine, Sphingomyelin, a-tocopherol, all-trans retinol, Astaxanthin and flavonoid. [0018]
  • 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, Phosphatidylcholine, Phosphatidylinositol, Phosphatidylserine, Phosphatidylethanolamine, Sphingomyelin, Cholesterol, Triglycerides, Monoglycerides, a-tocopherol, all-trans retinol, Astaxanthin, Canthaxanthin, β-carotene, flavonoid, Zinc, Selenium, sodium, potassium and calcium. [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, Phosphatidylcholine, Phosphatidylinositol, Phosphatidylserine, Phosphatidylethanolamine, Sphingomyelin, Cholesterol, Triglycerides, Monoglycerides, a-tocopherol, all-trans retinol, Astaxanthin, Canthaxanthin, β-carotene, Zinc and Selenium. [0020]
  • The diseases that can be treated and/or prevented by the method of the present invention are cardiovascular diseases, arthritis, skin cancer, diabetes, premenstrual syndrome and transdermal transport enhancement. [0021]
  • In accordance with the present invention there is also provided a composition for the treatment and/or prevention 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. [0022]
  • 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. [0023]
  • In accordance with the present invention, it is also provided the use of krill and/or marine oil for the manufacture of a medicament for the treatment and/or prevention and/or therapy of the previously mentioned diseases. [0024]
    • DETAILED DESCRIPTION OF THE INVENTION
  • 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. [0025]
  • 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 pacifica), 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. [0026]
  • The extraction process can be described as the following: [0027]
  • (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; [0028]
  • (b) Separating the liquid and the solid phases; [0029]
  • (c) Recovering a lipid rich fraction from the liquid phase obtained at step (b) by evaporation of the solvent present in the liquid phase; [0030]
  • (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; [0031]
  • (e) Separating the liquid and the solid phases; and [0032]
  • (f) Recovering a lipid rich fraction from the liquid phase obtained at step (e) by evaporation of the solvent present in the liquid phase. [0033]
  • The active components of the enzyme-free krill and/or marine oil extract are: [0034]
  • Lipids [0035]
  • 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]
  • In the preferred embodiment of the present invention, the Omega-3 are found in more than 30 g/100 g. [0041]
  • ii) Omega-6: i. Linoleic acid: >0.9 g/100 g [0042]
  • ii. Arachidonic acid: <0.45 g/100 g, preferably <0.6 g/100 g [0043]
  • iii) Omega-9: i. Oleic acid: >5 g/100 g [0044]
  • iv) palmitic acid: >10 g/100 g [0045]
  • v) palmitoleic acid: 0.08 g/100 g [0046]
  • vi) stearic acid: >0.5 g/100 g [0047]
  • Phospholipids [0048]
  • Phosphatidylcholine: >4.5 g/100 g [0049]
  • Phosphatidylinositol: >107 mg/100 g [0050]
  • Phosphatidylserine: >75 mg/100 g [0051]
  • Phosphatidylethanolamine: >0.5 g/100 g [0052]
  • Sphingomyelin: >107 mg/100 g [0053]
  • Neutral Lipids [0054]
  • Cholesterol: <3 g/100 g [0055]
  • Triglycerides: <55 g/100 g [0056]
  • Monoglycerides: >0.5 g/100 g [0057]
  • In another embodiment of the present invention, the neutral lipids of the krill and/or marine extract also comprises: [0058]
  • Diglycerides: >0.5 g/100 g [0059]
  • Antioxydants [0060]
  • α-tocopherol (vitamin E): >1.0 IU/100 g [0061]
  • all-trans retinol (vitamin A): >1500 IU/100 g [0062]
  • β-carotene: >3000 μg/100 ml [0063]
  • Pigments [0064]
  • Astaxanthin: >20 mg/100 g [0065]
  • Canthaxanthin: >2 mg/100 g [0066]
  • Metals [0067]
  • Zinc: >0.1 mg/100 g [0068]
  • Selenium: >0.1 mg/100 g [0069]
  • In another embodiment of the present invention, the krill and/or marine extract also comprises: [0070]
  • Flavonoids: >0.5 mg/100 g [0071]
  • Sodium: <500 mg/100 g [0072]
  • Calcium: >0.1 mg/100 g [0073]
  • Potassium: >50 mg/100 g [0074]
  • Aluminum: <8.5 mg/100 g [0075]
  • Protein: >4 g/100 g [0076]
  • Moisture and volatile matter: <0.8% [0077]
  • 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. [0078]
  • The oil has the following stability indexes: [0079]
  • Peroxide value: <0.1 (mEq/kg) [0080]
  • Oil Stability index: <0.1 after 50 hours at 97.8° C. [0081]
  • Saponification index: 70-180 [0082]
  • Iodine value:60-130% [0083]
  • The present invention will be more readily understood by referring to the following examples which are given to illustrate the invention rather than to limit its scope. [0084]
    • EXAMPLE 1 Cardiovascular Disease Prevention and/or Treatment
  • 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 inflammation 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 antithrombin 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) [0085]
  • 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. [0086]
  • 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. Recommended 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. [0087]
  • The patients were tested for LDL, HDL, Triglycerides, vital signs, CBC, SGOT/SGPT, γ-GT, ALP, Urea, Creatine, Glucose, K[0088] +, Na+, Ca2+ and total indirect bilirubin cholesterol before treatment and also at 2 months.
  • Table 1 is showing the results obtained from the previously described tests: [0089]
    TABLE 1
    Paired Samples Test
    Paired Differences
    95% Confidence
    Std. Interval of the
    Parameter Error Difference Sig. (2-
    tested Mean SD. Mean Lower Upper t-value df tailed)
    Cholesterol .4954 .55800 .15476 .1582 .8326 3.201 12 .008
    Triglycerides .3538 .54543 .15127 .0242 .6834 2.339 12 .037
    HDL −.2108 .29859 .08281 −.3912 −.0303 −2.545 12 .026
    LDL .2846 .47333 .13128 −.0014 .5706 2.168 12 .051
    Chol/HDL .3600 .53446 .14823 .0370 .6830 2.429 12 .032
  • 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 Cholesterol/HDL ratio decrease of 14%. [0090]
  • 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. [0091]
    • EXAMPLE 2 Arthritis Treatment
  • 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 Interleukin-8 and Interleukin-1 in human patients. Patients with a bleeding tendency or severe psychiatric disease were excluded from the study. [0092]
  • 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. [0093]
  • 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 consisting of 20% fat (less than 10% animal fat), 40% protein and 40% carbohydrates. [0094]
  • 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 including pain and stiffness, radiographic confirmation of illness prior to enrollment. It also include evidence of measurable symptoms of OA for at least 3 months prior to study enrollment requiring the use of acetaminophen, anti-inflammatory 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. [0095]
  • The Exclusion criteria were a severe osteoarthritis, unavoidable sustained use of NSAID's, aspirin or other medicines for anti-inflammatory use, use of topical analgesics 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 consumption exceeding 3 mixed drinks per day, concurrent medical/arthritic disease that could confound or interfere with the evaluation of pain, prior surgery (including arthroscopy) of either knee, a known “secondary” cause of osteoarthritis. [0096]
  • 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. Preliminary 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. [0097]
  • Results shown at Table 2 demonstrate the effect of an uptake of krill extract on the relief of arthritis. [0098]
    TABLE 2
    Frequency % Valid % Cumulative %
    No change 3 23.1 23.1 23.1
    Pain relief 10 76.9 76.9 100.0
    Total 13 100.0 100.0
  • This shows that ten out of 13 (76.9%) people reported a significant pain relief and improvement of flexibility of large joints (lower back, knees, shoulders) [0099]
    • EXAMPLE 3 Skin Cancer Prophylaxis
  • 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 phopholipid anti-carcinogenic effect. [0100]
  • 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. [0101]
  • 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 application. 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. [0102]
  • 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. [0103]
  • The mice were divided in six groups as follows: [0104]
  • Group A: fat-free chow with supplementation of fish oil (20% of total calories) [0105]
  • Group B: fat-free chow (100% of calories)+local application of fish oil 2 times per day [0106]
  • Group C: fat free chow with supplementation of fish oil (20% of total calories)+local application of soy oil 2 times per day [0107]
  • Group D: fat-free chow with supplementation of krill and/or marine oil (20% of total calories) [0108]
  • Group E: fat free chow (100% of calories)+local application of krill and/or marine oil 2 times per day [0109]
  • 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 [0110]
  • 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. [0111]
  • 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. [0112]
    TABLE 3
    Krill extract Oral uptake
    Cumulative
    Frequency Percent Valid Percent Percent
    Valid Benign 14 87.5 87.5 87.5
    Cancer 2 12.5 12.5 100.0
    Total 16 100.0 100.0
  • [0113]
    TABLE 4
    Control Oral uptake
    Cumulative
    Frequency Percent Valid Percent Percent
    Valid Benign 14 87.5 87.5 87.5
    Cancer 2 12.5 12.5 100.0
    Total 16 100.0 100.0
  • [0114]
    TABLE 5
    Krill extract topical uptake
    Cumulative
    Frequency Percent Valid Percent Percent
    Valid BENIGN 16 100.0 100.0 100.0
  • [0115]
    TABLE 6
    Control topical uptake
    Cumulative
    Frequency Percent Valid Percent Percent
    Valid BENIGN 5 31.3 31.3 31.3
    Cancer 11 68.8 68.8 100.0
    Total 16 100.0 100.0
  • [0116]
    TABLE 7
    Krill extract topical and oral uptake
    Cumulative
    Frequency Percent Valid Percent Percent
    Valid BENIGN 16 100.0 100.0 100.0
  • [0117]
    TABLE 8
    Control topical and oral uptake
    Cumulative
    Frequency Percent Valid Percent Percent
    Valid BENIGN 10 62.5 62.5 62.5
    Cancer 6 37.5 37.5 100.0
    Total 16 100.0 100.0
  • 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. [0118]
    • EXAMPLE 4 Transdermal Transport in Therapeutic Applications
  • Krill and/or marine oil enhances transdermal transportation as a substrate for dermatological topical therapeutic 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. [0119]
  • To evaluate the efficacy of krill and/or marine oil as a substrate for topical treatments and the speed of transdermal 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). [0120]
  • 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. [0121]
    • EXAMPLE 5 Transdermal Transport for Dermatological Topical Cosmetic Applications
  • 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. [0122]
  • 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-dermatological condition, bleeding tendency or severe psychiatric disease. [0123]
  • 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. [0124]
  • Table 9 shows results obtained on skin hydration following the method previously described. [0125]
    TABLE 9
    Changes in skin hydration
    Frequency % Valid % Cumulative %
    No change 4 30.8 30.8 30.8
    Hydration 9 69.2 69.2 100.0
    Total 13 100.0 100.0
  • The results of the pilot study after 2 months indicate 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. [0126]
  • 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: [0127]
  • Regeneration and distinctive anti-inflammatory effects [0128]
  • Improve blood irrigation [0129]
  • Increases the epidermis regeneration by increasing the rate of cell division and turnover [0130]
  • Accelerates the differentiation of keratin [0131]
  • Regenerates the collagen [0132]
  • Allows cells in the top layer of the skin, which are always being replaces, to mature more normally than untreated sun-damaged cells [0133]
  • Reduces the activation of enzymes that break down the proteins collagen and elastin that provide structural support for the skin. [0134]
  • 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. [0135]
    • EXAMPLE 6 Premenstrual Syndrome
  • 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. [0136]
  • All women enrolled reported noticeable emotional and/or physical discomfort 7 to 10 days prior to menstruation. 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. [0137]
  • 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). [0138]
    TABLE 10
    Frequency distribution of the effect of krill extract on
    premenstrual syndrome symptomatology
    PMS symptoms Frequency % Valid % Cumulative %
    No change 26.7 26.7 26.7
    Positive 73.3 73.3 100.0
    Total 100.0 100.0
    • EXAMPLE 7 Diabetes
  • 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. [0139]
    TABLE 11
    Variation in glucose in patients
    Paired Differences
    95%
    Confidence
    Para- Std. Interval Sig.
    meter Error of the t- (2-
    tested Mean SD. Mean Difference value df tailed)
    Glucose .5778 .60369 .20123 .1137-1.0418 2.871 8 .021
  • 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. [0140]
  • While the invention has been described in connection 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. [0141]

Claims (28)

1-83. (Cancelled)
84. A composition comprising an amount of krill oil effective for decreasing cholesterol, inhibiting platelet adhesion, inhibiting artery plaque formation, preventing hypertension, controlling arthritis symptoms, preventing skin cancer, enhancing transdermal transportation of dermatological topical therapeutic applications, enhancing transdermal transportation of dermatological cosmetic applications, reducing symptoms of premenstrual syndrome, or controlling 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, Docosahexanoic acid, Phosphatidylcholine, Phosphatidylinositol, Phosphatidylserine, Phosphatidylethanolamine, Sphingomyelin, 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, β-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 adhesion 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 applications, or enhancing transdermal transportation of dermatological 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 composition comprising an effective amount for controlling blood glucose level in a patient.
98. A method for decreasing cholesterol, inhibiting platelet adhesion, inhibiting artery plaque formation, preventing hypertension, controlling arthritis symptoms, preventing skin cancer, enhancing transdermal transportation of dermatological topical therapeutic applications, enhancing transdermal transportation of dermatological cosmetic applications, reducing symptoms of premenstrual syndrome, or controlling blood glucose level in a patient in need thereof, said method comprising administering an effective amount of the composition of claim 84 to said patient.
99. A method of claim 98, wherein said administering is effected orally.
100. A method of claim 99, wherein a quantity in a range of about 1 to almost 4.8 grams per day of krill oil is administered.
101. A method of claim 100, wherein said quantity is about 4.8 grams.
102. A method of claim 98, which is for decreasing cholesterol in a patient.
103. A method of claim 98, which is for inhibiting platelet adhesion and artery plaque formation in a patient.
104. A method of claim 98, which is for preventing hypertension in a patient.
105. A method of claim 98, which is for controlling arthritis symptoms in a patient.
106. A method of claim 105, wherein the arthritis is rheumatoid arthritis or osteoarthritis.
107. A method of claim 98, which is for preventing skin cancer in a patient.
108. A method of claim 98, which is for enhancing transdermal transportation of dermatological topical therapeutic applications, or enhancing transdermal transportation of dermatological cosmetic applications decreasing cholesterol in a patient.
109. A method of claim 98, which is for reducing symptoms of premenstrual syndrome in a patient.
110. A method of claim 98, which is for controlling blood glucose level in a patient.
US10/481,040 2001-06-18 2002-06-07 Krill and/or marine extracts for prevention and/or treatment of cardiovascular diseases arthritis, skin cancer diabetes, premenstrual syndrome and transdermal transport Abandoned US20040241249A1 (en)

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US11/640,235 US8057825B2 (en) 2001-06-18 2006-12-18 Krill extracts for treatment of cardiovascular diseases
US13/216,694 US20110305771A1 (en) 2001-06-18 2011-08-24 Krill Extracts for Treatment of Cardiovascular Diseases
US14/152,603 US20140199414A1 (en) 2001-06-18 2014-01-10 Krill and/or marine extracts for prevention and/or treatment of cardiovascular diseases, arthritis, skin cancer, diabetes, premenstrual syndrome and transdermal transport
US15/267,988 US20170224742A1 (en) 2001-06-18 2016-09-16 Phospholipid-containing marine extracts for treatment of cardiovascular diseases

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US10/481,040 US20040241249A1 (en) 2001-06-18 2002-06-07 Krill and/or marine extracts for prevention and/or treatment of cardiovascular diseases arthritis, skin cancer diabetes, premenstrual syndrome and transdermal transport

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US11/640,235 Expired - Fee Related US8057825B2 (en) 2001-06-18 2006-12-18 Krill extracts for treatment of cardiovascular diseases
US13/216,694 Abandoned US20110305771A1 (en) 2001-06-18 2011-08-24 Krill Extracts for Treatment of Cardiovascular Diseases
US14/152,603 Abandoned US20140199414A1 (en) 2001-06-18 2014-01-10 Krill and/or marine extracts for prevention and/or treatment of cardiovascular diseases, arthritis, skin cancer, diabetes, premenstrual syndrome and transdermal transport
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US15/267,988 Abandoned US20170224742A1 (en) 2001-06-18 2016-09-16 Phospholipid-containing marine extracts for treatment of cardiovascular diseases

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