WO2010068815A2 - Compositions and methods for treating cellular proliferative disorders - Google Patents

Compositions and methods for treating cellular proliferative disorders Download PDF

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WO2010068815A2
WO2010068815A2 PCT/US2009/067580 US2009067580W WO2010068815A2 WO 2010068815 A2 WO2010068815 A2 WO 2010068815A2 US 2009067580 W US2009067580 W US 2009067580W WO 2010068815 A2 WO2010068815 A2 WO 2010068815A2
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extract
cells
natural antioxidant
cell
fucoidan
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PCT/US2009/067580
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French (fr)
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WO2010068815A3 (en
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Hideaki Hagiwara
Kumiko Hagiwara
Takaaki Hagiwara
Yasunobu Takeshima
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Hihimsa Foundation
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/82Theaceae (Tea family), e.g. camellia
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/45Ericaceae or Vacciniaceae (Heath or Blueberry family), e.g. blueberry, cranberry or bilberry
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/73Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/73Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/736Prunus, e.g. plum, cherry, peach, apricot or almond
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/74Rubiaceae (Madder family)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/87Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/88Liliopsida (monocotyledons)
    • A61K36/889Arecaceae, Palmae or Palmaceae (Palm family), e.g. date or coconut palm or palmetto
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00Drugs for disorders of the cardiovascular system

Definitions

  • compositions and methods described herein relate generally to medicine and the treatment of cellular proliferative disorders, and more particularly, to compositions and methods for treating such disorders.
  • Cancer remains one of the most significant health problems world wide, and ranks second only to heart disease as a leading cause of death in the United States. Cancer, for the most part, involves uncontrolled proliferation and altered differentiation of the involved cells.
  • compositions of fucoidan and a natural antioxidant extract are also provided.
  • methods of treating a cellular proliferative disorder in a subject by administering to the subject a combination of fucoidan and a natural antioxidant extract.
  • the cellular proliferative disorder is cancer such as, but not limited to, melanoma, glioma, medulloblastoma, prostate cancer, esophageal cancer, lung cancer, breast cancer, ovarian cancer, testicular cancer, liver cancer, kidney cancer, renal cancer, spleen cancer, bladder cancer, colorectal and/or colon cancer, cervical cancer, pancreatic cancer, gall bladder cancer, stomach cancer, head and neck cancer, carcinoma, sarcoma, hepatoma, lymphoma, mycosis fungoides, leukemia, and a brain tumor.
  • cancer such as, but not limited to, melanoma, glioma, medulloblastoma, prostate cancer, esophageal cancer, lung cancer, breast cancer, ovarian cancer, testicular cancer, liver cancer, kidney cancer, renal cancer, spleen cancer, bladder cancer, colorectal and/or colon cancer, cervical cancer, pancreatic cancer, gall bladder cancer, stomach cancer, head
  • the cellular proliferative disorder is a neurodegenerative disorder such as, but not limited to, amyotrophic lateral sclerosis, Alzheimer's disease, Huntington's disease, Parkinson's disease, Schizophrenia, and Prion diseases.
  • the cellular proliferative disorder is a neovascular disorder such as, but not limited to, diabetic retinopathy, age-related macular degeneration, rheumatoid arthritis, osteoarthritis, glaucoma, keloids, corneal graft rejection, wound granularization, angiofibroma, Osier- Webber Syndrome, and myocardial angiogenesis.
  • the fucoidan and the natural antioxidant extract are administered simultaneously.
  • the fucoidan and the natural antioxidant extract are administered sequentially, such as, for example, the fucoidan is administered up to one, two, or three days prior to administration of the natural antioxidant extract.
  • the hyperproliferative cells are derived from a cellular proliferative disorder such as, but not limited to, melanoma, glioma, medulloblastoma, prostate cancer, esophageal cancer, lung cancer, breast cancer, ovarian cancer, testicular cancer, liver cancer, kidney cancer, renal cancer, spleen cancer, bladder cancer, colorectal and/or colon cancer, cervical cancer, pancreatic cancer, gall bladder cancer, stomach cancer, head and neck cancer, carcinoma, sarcoma, hepatoma, lymphoma, mycosis fungoides, leukemia, and a brain tumor.
  • a cellular proliferative disorder such as, but not limited to, melanoma, glioma, medulloblastoma, prostate cancer, esophageal cancer, lung cancer, breast cancer, ovarian cancer, testicular cancer, liver cancer, kidney cancer, renal cancer, spleen cancer, bladder cancer, colorectal and/or colon
  • hypoproliferative cells are derived from a cellular proliferative disorder such as, but not limited to, anemia and ischemia.
  • the fucoidan and the natural antioxidant extract are administered simultaneously.
  • the fucoidan and the natural antioxidant extract are administered sequentially, such as, for example, the fucoidan is administered up to one, two, or three days prior to administration of the natural antioxidant extract.
  • the hyperproliferative cells are derived from a cellular proliferative disorder such as, but not limited to, melanoma, glioma,
  • the method is performed in vivo. In another embodiment, the method is performed in vitro. In another embodiment, the fucoidan and the natural antioxidant extract are administered simultaneously. In another embodiment, the fucoidan and the natural antioxidant extract are administered sequentially, such as, for example, the fucoidan is administered up to one, two, or three days prior to administration of the natural antioxidant extract.
  • the method includes detecting inhibited cell proliferation in a sample of hyperproliferative cells as compared to cell proliferation in a corresponding untreated sample, thereby identifying a cell proliferative disorder amenable to treatment with fucoidan in combination with a natural antioxidant extract.
  • the method is performed in vivo. In another embodiment, the method is performed in vitro.
  • the natural antioxidant extract used in any of the aformentioned methods is a combination of one or more natural antioxidant extracts.
  • Exemplary natural antioxidant extracts include one or more extracts selected from acai berry extract, green tea extract, coffee bean extract, blueberry extract, grape extract, cherry extract, blackberry extract, cranberry extract, raspberry extract, strawberry extract, huckleberry extract, lemon extract, melon extract, kiwi extract, grapefruit extract, orange extract, apple extract, apricot extract, prune extract, watermelon extract, mangosteen extract, plum extract, date extract, banana extract, tomato extract, tumeric extract, broccoli extract, green barley leaf extract, red chili extract, carrot extract, and safran.
  • the sample of cells is any sample, including, for example, a tumor sample obtained by biopsy of a subject having the tumor, a tumor sample obtained by surgery (e.g., a surgical procedure to remove and/or debulk the tumor), or a sample of the subject's bodily fluid.
  • compositions comprising fucoidan and one or more natural antioxidant extracts.
  • such compositions further include a pharmaceutically and/or neutraceutically acceptable carrier.
  • the preparation is in the form of a tablet, capsule, granule, powder, suspension, emulsion, elixir or solution.
  • Figure 1 is an illustrative graphical diagram showing the results from a cell growth inhibition assay comparing concentrations of fucoidan and acai berry extract.
  • Figure 2 is an illustrative graphical diagram showing the results from a cell growth inhibition assay demonstrating synergy of inhibition with combinations of fucoidan and acai berry extract.
  • Figure 3 is an illustrative graphical diagram showing the results from a cell growth inhibition assay demonstrating synergy of inhibition with combinations of fucoidan/green tea extract and fucoidan/coffee bean extract.
  • Figure 4 is an illustrative graphical diagram showing the results from a cell growth inhibition assay demonstrating synergy of inhibition with combinations of fucoidan and blueberry extract.
  • Figure 5 is an illustrative graphical diagram showing the results from a negative control cell growth inhibition assay showing combinations of fucoidan and dried red grape extract.
  • Figure 6 is an illustrative graphical diagram showing the results from an apoptosis attenuation assay demonstrating a reduction in apoptosis with fucoidan.
  • Figure 7 is an illustrative graphical diagram showing the results from an apoptosis attenuation assay demonstrating a reduction in apoptosis with a natural antioxidant.
  • Figure 8 is an illustrative graphical diagram showing the results from an apoptosis attenuation assay demonstrating synergistic activity with fucoidan and green tea extract.
  • Figure 9 is an illustrative graphical diagram showing the results from an apoptosis attenuation assay demonstrating synergistic activity with fucoidan and green tea extract.
  • Fucoidan is a natural product that is obtained as an extract from brown algae such as nemacystus, undaria seaweeds, and sea tangles. Structurally, fucoidan is a sulfated polysaccharide whose primary constituent sugar is fucose. Fucoidan has become a desirable additive for food, cosmetic, and pharmaceutical preparations based on reports of the various pharmacologic properties of fucoidan. For example, fucoidan has been reported to have antitumor effects (Maruyama, F. et al. Kitasato Arch. Of Exp. Med., 60: 105-121 (1987), Ellouali, M.
  • compositions and methods capable of inhibiting unregulated growth and differentiation of cancer cells.
  • Described herein are formulations and methods of treating a variety of cellular proliferative disorders such as cancer. Described herein are methods for inhibiting growth of hyperproliferative cells, stimulating growth of hypoproliferative cells, and treating such disorders related thereto. Also provided are compositions of fucoidan and a natural antioxidant extract. [0024] As used in this specification and the appended claims, the singular forms "a”, “an”, and “the” include plural references unless the context clearly dictates otherwise.
  • references to “the method” includes one or more methods, and/or steps of the type described herein which will become apparent to those persons skilled in the art upon reading this disclosure and so forth. [0025] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.
  • Fucoidan is a sulfated polysaccharide, which is found in abundance in the slippery texture of Brown Algae.
  • the first isolation of fucoidan from marine algae was reported by Dr. Killing in 1913.
  • Algal fucoidan was found to be present in several orders of brown algae (Phaeophyceae), mainly Fucales and Laminariales, but also in Chordariales, Dictyotales, Dictyosiphonales, Ectocarpales and Scytosiphonales.
  • Fucoidan often called fucans, is composed mainly of sulfated L-fucose, but is also made of various proportions of different sugar residues such as fucose, galactose, xylose and uronic acids.
  • fucoidan is used to describe a sulfated complex polysaccharide (fucan) extracted from brown algae containing 20% to 60% of L-fucose.
  • Fucoidan is found in various species of brown seaweed including and not limited to kombu, limu moui, bladderwrack, wakame, mozuku, hijiki, Rhodophyceae such as common laver (Porphyra tenera), Gelidium cartiliagimeum and Gracilaria confervoides; Chlorophyceae such as Ulva lactuca; and Phaeophyceae such as Ecklonia cava, Eisenia arboria var.
  • algae refers to any of an organism classified into the Protista kingdom and is a chlorophyll-bearing organism that can survive in both salt- and freshwater, reproduce from a unicellular spore, and ranges in size from a single cell to giant kelp, including most kinds of seaweed.
  • brown algae refers to any organism classified into the Protista kingdom and further into the Phaeophyta division.
  • fucoidans have been known for some time to act as modulators of coagulation, as have other algal polysaccharides, fucoidans from brown algae were regarded only as a potential source of L-fucose for many years. Recently, a search for new biological modifiers has raised interest in sulfated polysaccharides. Fucoidans have been proposed as alternatives to the anti-coagulant Heparin, which is prepared from mammalian mucosa because, being of vegetable origin, fucoidans are less likely to contain infectious agents, such as viruses or prions.
  • fucoidans affect many cellular-biological and biochemical activities, such as anti-inflammation, anti- virus/parasites infection, anti-obesity, anti-cancer, immune enhancement, antithrombogenic, anti-complement effect, protection of vascular system, anti-he licobacter pylori, anti-ulcer, anti-renal stone formation, protection of liver function, beneficial for wound healing, anti- oxidative activities, anti-neuropathy, etc.
  • fucoidan has recently been demonstrated to mobilize hematopoetic stem cells, which contributes to regenerative medicine.
  • a combination of fucoidan and one or more natural antioxidant extracts provides improved therapeutic benefit (e.g., greater or more rapid reduction in cell proliferation) compared to the administration of fucoidan alone or one or more antioxidant extracts alone.
  • a combination of fucoidan and one or more natural antioxidant extracts provides improved therapeutic benefit (e.g., enhanced anti-apoptotic activity) compared to the administration of fucoidan alone or one or more antioxidant extracts alone.
  • a combination of fucoidan and one or more natural antioxidant extracts provides improved therapeutic benefit (e.g., increased apoptosis) compared to the administration of fucoidan alone or one or more antioxidant extracts alone.
  • administration of a combination of fucoidan and one or more natural antioxidant extracts produces a synergistic effect (e.g.., a more potent therapeutic effect) compared to treatment with one of the components alone.
  • acai berry extract enhances the activity of fucoidan on cancer cells by inhibiting cell growth and inducing apoptosis.
  • administration of a combination of fucoidan and one or more natural antioxidant extracts produces a synergistic effect (e.g., a more potent therapeutic effect) at lower doses of each component compared to treatment with one of the components alone.
  • a cell proliferative disorder described herein is caused by oxidative stress to a cell and/or tissue.
  • oxidative stress contributes to generation of free radicals (e.g., reactive oxygen species including and not limited to peroxide, superoxide or the like) that cause cellular and/or tissue injury including, for example, apoptosis and/or hyperproliferation.
  • free radicals e.g., reactive oxygen species including and not limited to peroxide, superoxide or the like
  • apoptosis that is induced by oxidative stress triggers a cascade of reactions in cells and/or tissues that lead to symptoms of neurodegenerative disorders including and not limited to Lou Gehrig's disease (aka MND or ALS), Parkinson's disease, Alzheimer's disease, Huntington's disease or any other neurodegenerative disorders described herein.
  • apoptosis that is induced by oxidative stress triggers a cascade of reactions in cells and/or tissues that lead to symptoms of certain cardiovascular disease.
  • oxidation of Low Density Lipids in the vascular endothelium is a precursor to plaque formation and subsequent manifestation of disorders of the vasculature, including defective angiogenesis, neovascular and/or cardiovascular disorders.
  • apoptosis that is induced by oxidative stress triggers a cascade of reactions in cells and/or tissues and plays a role in the ischemic cascade.
  • apoptosis that is induced by oxidative stress triggers a cascade of reactions in cells and/or tissues and leads to symptoms of chronic fatigue syndrome.
  • hyperproliferation that is induced by oxidative stress triggers a cascade of reactions in cells and/or tissues and leads to cancers including and not limited to melanoma, leukemia, or any other cancers described herein.
  • oxidative stress-induced and/or free radical-induced apoptosis and/or hyperproliferation of cells underlies the etiology of a wide range of disorders including any disorders described herein.
  • administration of a combination of fucoidan and one or more antioxidant extracts produces a synergistic effect that reduces cellular oxidative stress and/or generation of free radicals.
  • administration of a combination of fucoidan and one or more antioxidant extracts produces a synergistic effect that reduces or delays the progression of cell proliferative disorders.
  • administration of a combination of fucoidan and one or more antioxidant extracts produces a synergistic anti-apoptotic effect.
  • cell proliferative disorder refers to a hyperproliferative disorder or a hypoproliferative disorder.
  • a “hyperproliferative disorder” refers to any state or condition that results in an abnormal increase in the number of cells as a result of cell growth and cell division, as compared to the number of cells resulting from cell growth or cell division of a corresponding normal cell.
  • a “hypoproliferative disorder” refers to any state or condition that results in the destruction, shedding, or inadequate growth/proliferation or excessive cell death (e.g., apoptosis or necrosis) of a particular cell type, as compared to the number of cells resulting from cell growth, cell division, or cell death of a corresponding normal cell.
  • a “hyperproliferative cell” refers to any cell that undergoes an abnormal increase in the number of cells as a result of cell growth and cell division, as compared to the number of cells resulting from cell growth or cell division of a corresponding normal cell.
  • a “hypoproliferative cell” refers to any cell that undergoes increased destruction, increased shedding, decreased growth/proliferation, or excessive cell death, as compared to a corresponding normal cell.
  • a cell proliferative disorder as described herein is a neoplasm.
  • neoplasms are either benign or malignant.
  • the term “neoplasm” refers to a new, abnormal growth of cells or a growth of abnormal cells that reproduce more aggressively than normal.
  • a neoplasm creates an unstructured mass (a tumor) which can be either benign or malignant.
  • the term “benign” refers to a tumor that is noncancerous, e.g., its cells do not invade surrounding tissues or metastasize to distant sites.
  • malignant refers to a tumor that is cancerous, metastastic, invades contiguous tissue or is no longer under normal cellular growth control.
  • the cell proliferative disorder is cancer.
  • Exemplary cancers that can be treated by the methods and compositions described herein include, but are not limited to, melanoma, glioma, teratoma, medulloblastoma, seminoma, prostate cancer, esophageal cancer, lung cancer, breast cancer, ovarian cancer, testicular cancer, liver cancer, kidney/renal cancer, cervical cancer, pancreatic cancer, gall bladder cancer, stomach cancer, spleen cancer, bladder cancer, colorectal cancer and/or colon cancer, head and neck cancer, heart cancer, carcinoma, sarcoma, hepatoma, lymphoma, mycosis fungoides, leukemia, and brain tumors.
  • a cell proliferative disorder as described herein is a neurodegenerative disorder.
  • the term "neurodegenerative disorder” as used herein includes any disorder in which cells of the retina, brain and spinal cord are lost.
  • neurodegenerative disorders that can be treated by the methods and compositions described herein include, but are not limited to, amyotrophic lateral sclerosis, Alzheimer's disease, Huntington's disease, Parkinson's disease, Schizophrenia, alcoholism, Alexander's disease, Alper's disease, Ataxia telangiectasia, Batten disease, bovine spongiform encephalopathy (BSE), canavan disease, cockayne syndrome, corticobasal degeneration, Creutzfe lft- Jakob disease, HIV-associated dementia, ischemia, Kennedy's disease, Krabbe's disease, Lewy body dementia, spinocerebellar ataxia, multiple sclerosis, multiple system atrophy, narcolepsy, neuroborreliosis, palizaeus-merzbacher disease, Pick's disease, primary lateral sclerosis, Prion diseases, Refsum's disease, Schildr's disease, Sanhoff s disease, subacute combined degeneration of spinal cord
  • a cell proliferative disorder as described herein is a neovascular disorder and/or a neovascularization-associated disorder.
  • neovascular disorder refers to any disorder characterized by the formation of functional microvascular networks with red blood cell perfusion.
  • exemplary neovascular disorders that can be treated by the methods and compositions described herein include, but are not limited to, solid tumors, diabetic retinopathy, age-related macular degeneration, rheumatoid arthritis, osteoarthritis, glaucoma, keloids, corneal graft rejection, wound granularization, angiofibroma, Osier- Webber Syndrome, myocardial angiogenesis, Ataxia Telangiectasia, vasoagglutination, vascular deformation, endotheioma, hypertrophy, progeria, psoriasis, sarcoma, calorification, arthritis, emphysema, bronchitis, obesity, and cataracts.
  • a cell proliferative disorder as described herein is a hypoproliferative disorder.
  • Exemplary hypoproliferative disorder is
  • corresponding normal cells refers to cells, or a sample from a subject that is free of the cell proliferative disorder being treated and is from the same organ and of the same type as the cells being examined.
  • the corresponding normal cells comprise a sample of cells obtained from a healthy individual that does not have a cell proliferative disorder.
  • Such corresponding normal cells can, but need not be, from an individual that is age -matched and/or of the same sex as the individual providing the cells being examined.
  • parenteral administration and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually orally or by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
  • systemic administration means the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the subject's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.
  • terapéuticaally effective amount or “effective amount” means the amount of a compound or composition that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician.
  • the term “ameliorating” or “treating” means that the clinical signs and/or the symptoms associated with a particular cell proliferative disorder are lessened as a result of the actions performed.
  • the signs or symptoms to be monitored will be characteristic of a particular cell proliferative disorder and will be well known to the skilled clinician, as will the methods for monitoring the signs and conditions.
  • subject refers to any individual or patient to which the subject methods are performed. Generally the subject is human, although as will be appreciated by those in the art, the subject is an animal. Thus other animals, including mammals such as rodents (including mice, rats, hamsters and guinea pigs), cats, dogs, rabbits, farm animals including cows, horses, goats, sheep, pigs, etc., and primates (including monkeys, chimpanzees, orangutans and gorillas) are included within the definition of subject.
  • rodents including mice, rats, hamsters and guinea pigs
  • cats dogs, rabbits, farm animals including cows, horses, goats, sheep, pigs, etc.
  • primates including monkeys, chimpanzees, orangutans and gorillas
  • methods of inhibiting growth of a hyperproliferative cell includes contacting the cell with fucoidan in combination with one or more natural antioxidant extracts.
  • provided herein are methods of ameliorating hyperproliferative cells in a subject.
  • the method includes administering to the subject in need of such treatment a therapeutically effective dosage of fucoidan in combination with one or more natural antioxidant extracts.
  • methods of stimulating growth of a hypoproliferative cell The method includes contacting the cell with fucoidan in combination with one or more natural antioxidant extracts.
  • methods of ameliorating hypoproliferative cells in a subject The method includes administering to the subject in need of such treatment a therapeutically effective dosage of fucoidan in combination with one or more natural antioxidant extracts.
  • a "natural antioxidant extract” refers to the extract of any organic substance that is capable of counteracting the damaging effects of oxidation in animal tissues.
  • exemplary natural antioxidant extracts include, but are not limited to, acai berry extract, green tea extract, coffee bean extract, and blueberry extract.
  • Additional organic substances that provide an extract useful in the methods and compositions described herein include, but are not limited to, oranges, apples, tomatoes, pomegranates, lemons, pears, grapes, blueberries, cherries, blackberries, cranberries, raspberries, strawberries, huckleberries, lemons, melons, kiwis, grapefruits, oranges, apricots, prunes, watermelons, mangosteens, plums, dates, bananas, broccoli, green barley leaves, red chili, carrot, tumeric and safran.
  • one or more of the natural antioxidant extracts contain catechins.
  • Catechins are polyphenolic antioxidant plant metabolites.
  • All methods further include the step of bringing the active ingredient(s) (e.g., fucoidan alone or in combination with one or more natural antioxidant extracts) into association with a pharmaceutically and/or neutraceutically acceptable carrier, which constitutes one or more accessory ingredients.
  • the active ingredient(s) e.g., fucoidan alone or in combination with one or more natural antioxidant extracts
  • a pharmaceutically and/or neutraceutically acceptable carrier which constitutes one or more accessory ingredients.
  • the compositions for use in treating subjects having one or more cell proliferative disorders includes as the active constituent a therapeutically effective amount of fucoidan in combination with one or more natural antioxidant extracts together with a pharmaceutically and/or neutraceutically acceptable carrier, diluent of excipients.
  • compositions for administration to a subject include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil or injectable organic esters.
  • a pharmaceutically and/or neutraceutically acceptable carrier can contain physiologically acceptable compounds that act, for example, to stabilize or to increase the absorption of the conjugate.
  • physiologically acceptable compounds include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients.
  • composition also can contain a second (or more) compound(s) such as a diagnostic reagent, nutritional substance, toxin, or therapeutic agent, for example, a cancer chemotherapeutic agent and/or vitamin(s).
  • a second (or more) compound(s) such as a diagnostic reagent, nutritional substance, toxin, or therapeutic agent, for example, a cancer chemotherapeutic agent and/or vitamin(s).
  • fucoidan is administered in powder, liquid, or solid form.
  • the natural antioxidant extract is administered in powder, liquid, or solid form.
  • the fucoidan and the natural antioxidant extract are administered in identical forms (e.g., powder form).
  • the fucoidan and the natural antioxidant extract are administered in different forms.
  • the fucoidan is administered as a tablet or a capsule containing a powdered form thereof, and the natural antioxidant extract is administered in liquid form (e.g., in juice or mixed in water).
  • Formulations described herein that are suitable for oral administration are presented as discrete units such as capsules, cachets, tablets or lozenges, each containing a predetermined amount of the active compound in the form of a powder or granules; or as a suspension of the active compound in an aqueous liquid or non-aqueous liquid such as a syrup, an elixir, an emulsion or a draught.
  • aqueous liquid or non-aqueous liquid such as a syrup, an elixir, an emulsion or a draught.
  • Such a medicament is also useful for inhibiting growth of hyperproliferative cells and/or ameliorating symptoms associated with hyperproliferative cells in a subject.
  • a medicament is also useful for stimulating growth of hypoproliferative cells and/or ameliorating symptoms associated with hypoproliferative cells in a subject.
  • the methods described herein are further combined with other compounds, compositions or therapeutic regimens that are known to inhibit angiogenesis, neovascularization or excessive cellular proliferation such as that which occurs in cancer.
  • Such therapies include, but are not limited to chemotherapy, radiation therapy and the administration of anti-angiogenic compounds.
  • the total amount of a compound or composition to be administered in practicing a method described herein is administered to a subject as a single dose, either as a bolus or by infusion over a relatively short period of time, or can be administered using a fractionated treatment protocol, in which multiple doses are administered over a prolonged period of time (e.g., once daily, twice daily, etc.).
  • a fractionated treatment protocol in which multiple doses are administered over a prolonged period of time (e.g., once daily, twice daily, etc.).
  • the amount of fucoidan and the amount of natural antioxidant extract to treat cell proliferative disorders as herein defined depends on many factors including the age and general health of the subject as well as the route of administration and the number of treatments to be administered. In view of these factors, the skilled artisan would adjust the particular doses and concentrations as necessary.
  • the formulation of the composition and the routes and frequency of administration are determined, initially, using Phase I and Phase II clinical trials.
  • the methods described herein include an intervalled or sequential treatment regimen. It was observed that accumulation of fucoidan in a subject allows for administration of fucoidan followed by administration of the natural antioxidant extract within 2-5 days of administering the fucoidan. As such, in one embodiment, the fucoidan and the natural antioxidant extract are administered sequentially. For example, in one embodiment, the fucoidan is administered up to three days prior to administering the natural antioxidant extract. In another exemplary embodiment, the fucoidan is administered one or two days prior to administering the natural antioxidant extract. In another embodiment, the fucoidan and the natural antioxidant extract are administered simultaneously.
  • the ratio of fucoidan to natural antioxidant extract is anywhere from 5: 1 to 1 :5.
  • the ratio of fucoidan to natural antioxidant extract is 2: 1, 1 :2, or 1 : 1.
  • the method can be performed, for example, by contacting a sample of cells to be treated with a composition described herein and determining a change in the total cell number as a result of the contact, as compared to the total cell number of an untreated cell.
  • Detection of decreased cell number (i.e., inhibition of cell proliferation) in the hyperproliferative cells as compared to the untreated cells indicates that the cells can benefit from treatment.
  • detection of increased cell number (i.e., stimulation of cell proliferation) in the hypoproliferative cells as compared to the untreated cells indicates that the cells can benefit from treatment.
  • the methods described herein are useful for providing a means for practicing personalized medicine, wherein treatment is tailored to a subject based on the particular characteristics of the hyperproliferative or hypoproliferative cells in the subject.
  • the sample of cells examined according to the present method can be obtained from the subject to be treated, or can be cells of an established cancer cell line or known hyperproliferative and/or hypoproliferative cells of the same type as that of the subject.
  • the established cell line can be one of a panel of such cell lines, wherein the panel can include different cell lines of the same type of disease and/or different cell lines of different diseases associated with hyperproliferation or hypoproliferation.
  • Such a panel of cell lines can be useful, for example, to practice the present method when only a small number of cells can be obtained from the subject to be treated, thus providing a surrogate sample of the subject's cells, and also can be useful to include as control samples in practicing the present methods.
  • the methods described herein are repeated on a regular basis to evaluate whether the hyperproliferative or hypoproliferative cells in the subject begin to show resistance to the therapy.
  • the results obtained from successive assays are used to show the efficacy of treatment over a period ranging from several days to months.
  • the invention is also directed to methods for monitoring a therapeutic regimen for treating a subject having a cell proliferative disorder. A comparison of the total cell number prior to and during therapy indicates the efficacy of the therapy. Therefore, one skilled in the art will be able to recognize and adjust the therapeutic approach as needed.
  • the methods described herein are adaptable to a high throughput format, thus allowing the examination of a plurality (i.e., 2, 3, 4, or more) of cell samples and/or compositions, which independently can be the same or different, in parallel.
  • a high throughput format provides numerous advantages, including that various compositions can be tested on several samples of cells from a single subject, thus allowing, for example, for the identification of a particularly effective concentration of a particular ingredient of the composition to be administered to the subject, or for the identification of a particularly effective natural antioxidant extract, or combination thereof, to be administered to the subject in combination with fucoidan.
  • a high throughput format allows for the examination of two, three, four, etc., different extracts, alone or in combination, on the hyperproliferative cells of a subject such that the best (most effective) extract or combination of extracts can be used for a therapeutic procedure.
  • a high throughput format allows, for example, control samples (positive controls and or negative controls) to be run in parallel with test samples, including, for example, samples of cells known to be effectively treated with an extract being tested.
  • the methods can be performed on a solid support (e.g., a microtiter plate, a silicon wafer, or a glass slide), wherein samples to be contacted with an extract or combination thereof are positioned such that each is delineated from each other (e.g., in wells). Any number of samples (e.g., 96, 1024, 10,000, 100,000, or more) can be examined in parallel using such a method, depending on the particular support used.
  • a solid support e.g., a microtiter plate, a silicon wafer, or a glass slide
  • each sample in the array can be defined by its position (e.g., using an x-y axis), thus providing an "address" for each sample.
  • An advantage of using an addressable array format is that the method can be automated, in whole or in part, such that cell samples, reagents, test agents, and the like, can be dispensed to (or removed from) specified positions at desired times, and samples (or aliquots) can be monitored, for example, for decreased cell number.
  • Example I Tumor Suppressive and Apoptosis Activity of Acai and Fucoidan [0061] Cell line and culture.
  • the human lymphoma, HS-Sultan was purchased from the ATCC.
  • HS-Sultan was cultured in RPMI 1640 medium (Mediatech Inc.) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and Penicillin- Streptomycine solution (Mediatech
  • Fucoidan samples are dissolved in PBS (Mediatech Inc.) at a concentration of 50 mg/mL by incubating at 50 0 C for 2 hrs. If the solutions contained insoluble substances, the solutions were subjected to centrifugation at 12,000 rpm for 10 min to remove such insoluble substances. The sample solutions were then sterilized by 0.2 or 0.45 ⁇ m syringe filters (VWR).
  • VWR 0.45 ⁇ m syringe filters
  • WST-8 (20 ⁇ L) is added for the last 4 hrs of incubation, and the absorbance at 450nm is measured using an automated microplate reader. Measuring the mitochondrial dehydrogenase cleavage of WST- 8 to formazan dye indicates the level of proliferation.
  • Apoptosis assay (Caspase-3 assay). Cells were suspended at a final concentration of 1x10 5 cells/mL in RPMI1640 with 10% FBS and Penicillin-Streptomycin solution in a 96-well microculture plate in triplicate. Fucoidan was then added to each well at various concentrations, and incubated for 24 hrs. The activation of caspase-3 was analyzed using a caspase-3 assay kit according to the manufacturer's instructions.
  • This example demonstrates the synergistic tumor suppressive activity and apoptosis enhancing activity of fucoidan (Cladosiphon okamuranus) in combination with acai berry extract.
  • HS-Sultan cells were contacted with different concentrations (i.e., 0-2 mg/mL) of acai berry extract alone and in the presence of 1 -3 mg/mL fucoidan.
  • the cell number of Control was 71OxIO 3 cells/mL.
  • This example demonstrates the synergistic tumor suppressive activity and apoptosis enhancing activity of fucoidan in combination with extracts from other natural antioxidant containing substances.
  • HS-Sultan cells were contacted with various concentrations of green tea extract, coffee bean extract, and 1 mg/mL fucoidan in combination with each of those substances. In each instance, the addition of 1 mg/mL fucoidan showed a significant synergistic activity in cell growth inhibition.
  • HS-Sultan cells were contacted with various concentrations of blueberry extract to determine concentrations that inhibit cell growth and induce apoptosis. The addition of 1 mg/mL fucoidan showed a significant synergistic activity in cell growth inhibition.
  • dried red grape was used as a negative control in the cell growth inhibition assay on HS-Sultan. The addition of 1 mg/mL fucoidan showed no significant synergistic activity in cell growth inhibition over dried red grape extract alone.
  • This example demonstrates the apoptosis attenuation activity of Fucoidan (Cladosiphon okamurae) on ethanol-induced apoptosis of HS-Sultan cells.
  • HS-Sultan was obtained and prepared as described in Example I.
  • Apoptosis Attenuation Assay The attenuation effects of fucoidan (Cladosiphon okamuranus) on ethanol (EtOH)-induced apoptosis of HS-Sultan cells were examined.
  • the cell number of Control was 675xlO 3 cells/mL.
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CN110494148A (zh) * 2017-02-09 2019-11-22 安特瑞珍有限公司 将人参/红参及海参复合提取物作为有效成分的布鲁赫膜功能下降相关疾病预防或治疗用组合物
CN110494148B (zh) * 2017-02-09 2023-10-27 安特瑞珍有限公司 人参/红参及海参复合提取物作为有效成分的布鲁赫膜功能下降相关疾病预防或治疗用组合物

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