US20120148683A1 - Immune stimulating composition comprising an extract of aronia sp. in combination with selenium and/or zinc - Google Patents
Immune stimulating composition comprising an extract of aronia sp. in combination with selenium and/or zinc Download PDFInfo
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- US20120148683A1 US20120148683A1 US13/320,021 US201013320021A US2012148683A1 US 20120148683 A1 US20120148683 A1 US 20120148683A1 US 201013320021 A US201013320021 A US 201013320021A US 2012148683 A1 US2012148683 A1 US 2012148683A1
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Definitions
- the present invention relates to an immune stimulating agent, and relates in particular to a composition comprising Aronia extract and selenium for stimulating the immune system, and pharmaceutical or nutraceutical compositions comprising the same.
- Aronia is a deciduous shrub which belongs to the plant family Rosaceae. Within the genus Aronia several species and hybrids are known, such as Aronia melanocarpa (black chokeberry), Aronia arbutifolia (red chokeberry), and Aronia x prunifolia (purple chokeberry). The Aronia fruits are small pomes and the juice of it is known to be astringent and to have a high content of vitamin C and other antioxidants.
- Aronia is used as a coloring agent for beverages and other food products, as disclosed e.g. in U.S. Pat. No. 6,703,056, WO 2005 058052, and WO 2006 138419. Beside this, the Aronia fruits have awaken interest due to their antioxidative qualities.
- Aronia melanocarpa is known to have high contents of phenolic compounds, especially anthocyanins, which are mainly located in the skin of the berries for UV protection of the pulp and the seeds. These anthocyanins give Aronia melanocarpa an extraordinary antioxidative strength.
- a parameter showing the capacity of a compound or an extract to deal with oxidative stress is the oxygen radical absorbance capacity or ORAC.
- Aronia shows high ORAC values and is therefore also used/added as a functional food or nutraceutical.
- U.S. 2007 0020358 discloses a sports drink concentrate which may include Aronia as a nutraceutical ingredient.
- a functional sweetener composition comprising an Aronia extract as antioxidant is disclosed in WO 2007 06 1900, and WO 2007 076857 teaches a chewing gum composition containing Aronia which is used as a source of antioxidants.
- Aronia or compounds thereof may be used for the preparation of pharmaceutical compositions or nutraceutical compositions and food supplements.
- DE 10 2004 029 887 Al discloses a dietary supplement on the basis of pomace comprising an extract of the skin of Aronia fruits which dietary supplement is described to have a high radical protection factor.
- DE 10 2004 052 882 A1 teaches a dietary supplement on the basis of a specific selection of fruit and legume concentrates, which comprises inter alia an Aronia fruit juice concentrate. Said dietary supplement is described to strengthen the immune system, wherein it is set forth that the combination of ingredients synergistically influences the impact of the ingredients.
- WO 2001 015553 teaches that food supplements containing Aronia fruit extract may be used to inhibit an inflammation mediated by cyclooxygenase, i.e. it is disclosed that Aronia extracts have an anti-inflammatory activity due to an inhibition of COX-2 activity preferentially over COK-1 activity.
- WO 2002 005827 teaches the use of Aronia extract for the preparation of a medicament for the treatment of brain cancer and further discloses a nutraceutical composition for the treatment of cancer comprising one or more flavonoids from Aronia and the micronutrients selenium and zinc.
- Micronutrients are needed and involved, e.g. as cofactors for enzymes, in a plurality of metabolism processes and are therefore important ingredients in food. Accordingly, and as taught by the prior art, micronutrients may be added to different kinds of food for example to functional food in order to achieve or improve certain physiological functions.
- the micronutrients selenium and zinc affect beside other physiological functions the immune status.
- Selenium is known to be required for a proper activity of neutrophils, macrophages, NK cells, T lymphocytes and some other immune mechanisms and an insufficient supply of selenium to the human body may result in a weakened immune system.
- Zinc is known to be an important ion for enzymes, proteins and transcription factors, and is therefore involved in many functions of the body also including immune responses.
- composition comprising an Aronia extract in an amount of at least about 10 ⁇ g/mg and selenium in an amount of from about 0.0001 ⁇ g/mg to about 2.0 ⁇ g/mg.
- selenium is added to the Aronia extract.
- the amount of selenium or the combination of selenium and zinc is beyond the natural fluctuation range in an Aronia extract.
- composition further comprises additional zinc, preferably in a concentration from about 0.0001 mg/mg to about 0.5 mg/mg.
- composition further comprises at least one phai inaceutically or nutraceutically acceptable excipient or a mixture thereof.
- composition is formulated as a pharmaceutical composition, preferably designed for oral, such as peroral, buccal, sublingual, mucosal, or topical administration.
- the composition is formulated as a nutraceutical or supplementary neutraceutical composition, preferably in the form of an ingestible solution or suspension, comprising the active ingredient in dissolved or suspended form, or as a solid administration form, such as in the form of tablets, capsules, dragees, or added as a food supplement to daily food products.
- composition according to the present invention is for the preparation of a medicament, preferably for the treatment of inflammatory diseases or conditions, bacterial infections, fungal infections, viral infections, parasitic infections and combinations thereof.
- the inflammatory disease or condition is preferably associated with autoimmune diseases, inflammatory bowel disease, multiple sclerosis, scleroderma, and graft-versus-host disease.
- FIGS. 1 and 2 shows evaluation of the optimal concentration of Aronia extract on macrophage activity of macrophages of the mouse leukaemic monocyte macrophage cell line RAW 264.7. It may be seen that concentrations above approx. 40 ⁇ g/ml decrease macrophage activity.
- FIG. 3 shows the immune stimulatory activity of selenium employing macrophages of the cell line RAW 264.7. It may be seen that concentrations above approx. 8 ⁇ g/ml decrease macrophage activity.
- FIG. 4 shows the immune stimulating effect of a composition according to the present invention by means of the activity of macrophages of the cell line RAW 264.7 in dependence from the added selenium concentration.
- the composition comprises an Aronia extract in a concentration of 4.875 ⁇ g/ml.
- the present invention discloses a composition which comprises a combination of an Aronia extract and the micronutrient selenium.
- the composition may also comprise zinc as further micronutrient, and is designed/ used for stimulating the immune system.
- the Aronia extract originates from one Aronia species which is pharmaceutically and nutraceutically acceptable or is a mixture of different pharmaceutically/ nutraceutically acceptable Aronia species.
- Aronia species that may be used include, but are not limited to A. melanocarpa, A. arbutifolia , and A. x prunifolia .
- Aronia melanocarpa is used.
- the Aronia extract may be derived from the whole fruit or from parts of the fruit.
- the extract may be produced on the base of the pomace of the fruits.
- the Aronia extracts may be obtained according to any suitable method known by a person skilled in the art.
- the extract may be based on fresh fruits or dried fruits or fruit juices. If desired, one or more particular active compounds may be selectively enriched in the extract.
- An Aronia extract may be obtained to the general knowledge of the skilled person. Suitable ways for preparing an Aronia extract are for example disclosed in DE 102004029887 and DE 102005046474. It will be, however, appreciated that in principle each preparation of plant extracts may be modified according to general knowledge in order to obtain Aronia extracts suitable for the present invention.
- General extraction methods for isolating plant material is for example disclosed in FR 2892933 and Ljubuncic P. et al. ( J. Ethnopharmacol ., Oct 3, 101(1-3) (2005), 153-61).
- the Aronia extract may be prepared using whole fruits as starting material.
- the fruits are washed with water of a temperature between 30 and 50° C., preferably of 40° C. and afterwards enzymatically treated with e.g. an inulinase, preferably fructozyme, for partial or complete cell disruption.
- Suitable enzymes as well as reaction conditions are well known to the skilled person.
- the residue obtained after pressing is extracted with aqueous potassium sulfite solution (approx. 10-30 w/v) for 1 to 3 hours at room temperature (approx. 21-24° C.).
- the extract is treated with pectin degrading enzymes, such as pectinex or pectinol, at a pH of approx. 3-4, preferably pH 3, for 0.5-2 hours.
- the extract is distilled under vacuum at a temperature of around 40-58° C., preferably 45-50° C., yielding Aronia extract having a residual water content of approx. 30-40 w/w.
- the Aronia extract may be prepared by means of an ethanol extraction or it may be prepared as a dry extract.
- the composition according to the present invention comprises the Aronia extract in a concentration from at least about 10 ⁇ g/mg to about 990 ⁇ g/mg, preferably from about 20 ⁇ g/mg to about 950 ⁇ g/mg. Even more preferred from about 40 ⁇ g/mg to about 900 ⁇ g/mg. Still more preferred is a concentration in the range from about 50 ⁇ g/mg to about 800 ⁇ g/mg. Still even more preferred is a concentration from about 60 ⁇ g/mg to about 800 ⁇ g/mg. Most preferably the Aronia extract is concentrated in a range from about 80 ⁇ g/mg to about 800 ⁇ g/mg.
- a fluid composition of the present invention comprises Aronia extract in a concentration of about 10 ⁇ g/mg to about 200 ⁇ g/mg, more preferred about 60 ⁇ g/mg to about 150 ⁇ g/mg and most preferred about 80 ⁇ g/mg.
- a solid or dried composition of the present invention comprises Aronia extract in a concentration of about 100 ⁇ g/mg to about 990 ⁇ g/mg, more preferred from about 300 ⁇ g/mg to about 800 ⁇ g/mg, even more preferred from about 400 ⁇ g/mg to about 700 ⁇ g/mg and most preferred about 600 ⁇ g/mg.
- the values given are intended to mean the final concentration of the Aronia extract in the composition according to the present invention, i.e. the concentration with which the extract is administered to a patient or a subject in need thereof.
- the composition also comprises the micronutrient selenium.
- the composition comprises Aronia extract and a combination of the micronutrients selenium and zinc.
- a micronutrient is intended to mean that additional amounts of said micronutrient(s) is/are added in addition to the micronutrients which may be already present in the Aronia extract.
- the Aronia extract particularly from the species Aronia melanocarpa , may contain natural amounts of 4 mg/kg zinc and about 20 ⁇ g/kg selenium. It will be appreciated that these natural amounts may vary dependent on the fruits specifies and growth conditions.
- micronutrient(s) selenium or selenium and zinc may be provided for example, but not limited thereto as inorganic salts, such as chlorides, sulfates, and the like, as organic salts and other bioavailable forms, such as amino acid chelates or combinations thereof.
- inorganic salts such as chlorides, sulfates, and the like
- organic salts and other bioavailable forms such as amino acid chelates or combinations thereof.
- the terms “selenium” and “zinc” are intended to comprise any compound containing selenium and zinc, respectively, including salts, complexes or other forms of said micronutrients, also including elemental selenium or zinc. Acceptable forms of selenium and zinc are well known to the skilled person. In case of given concentrations or ranges of concentrations of said micronutrients said values are intended to indicate the concentration of selenium or the selenium ion, and the concentration of zinc or the zinc ion, respectively.
- selenium may be provided as selenomethionine, elemental selenium, various selenium salts, such selenium chloride, and selenium yeast or combinations thereof According to a preferred embodiment of the present invention selenium is provided as selenium yeast or sodium selenite/ sodium selenate.
- zinc may be provided as zinc acetate, zinc ascorbate, zinc chloride, zinc citrate, zinc gluconate, zinc lactate, zinc sulphate, amino acid chelated zinc, or combinations thereof.
- zinc is provided as zinc chloride.
- composition according to the present invention may be provided in liquid form or in dried/solid form, e.g. in a lyophilized form. According to a preferred embodiment of the present invention the composition is provided in a sterile form.
- a composition comprising an Aronia extract and additionally added selenium or additionally added selenium and zinc has an enhanced immune stimulating effect in comparison to the single compounds of the composition according to the present invention.
- the beneficial immune stimulating effect of the composition according to the present invention is based on a synergistic effect of the complex mixture of compounds comprised in the Aronia extract in combination with the additionally added selenium.
- a further synergistic effect may be achieved when adding zinc to the composition comprising Aronia extract and selenium.
- composition according to the present invention has been shown to increase the macrophage activity.
- Macrophages belong to the innate immune system which is e.g. responsible for destroying virus infected cells, parasites and tumour cells. Macrophages represent matured monocytes, which in turn belong to the white blood cells. Macrophages phagocytose and digest invading microorganisms, even protozoa, debris, foreign bodies as well as damaged, dead and senescent cells.
- composition according to the present invention can be used to stimulate the immune system.
- the composition according to the present invention may be used for the preparation of a medicament for the treatment of a variety of diseases or conditions which are based on a reduced activity of the immune system, or which may be prevented or treated by stimulating the immune system.
- diseases or conditions may be treated or prevented that are based on a reduced macrophage activity or which may be prevented or treated by enhancing the macrophage activity.
- treatment is intended to mean prophylactic or therapeutic treatment, wherein the therapeutic treatment may be palliative or curative.
- composition according to the present invention is characterized by an immune stimulating activity it may be also used for the preparation of a medicament designed for adjuvant therapy during the treatment of diseases which treatment is beneficially affected by stimulating the immune system. Furthermore, the composition according to the present invention may also be used for the preparation of a dietary supplement for supporting the immune system.
- the composition is designed for the treatment of inflammatory diseases or conditions, bacterial infections, fungal infections, viral infections, parasitic infections and combinations thereof.
- composition may be used for alleviate or improving any disorder or disease including but not limited to septic shock, haemodynamic shock, sepsis syndrome, post ischaemic reperfusion injury, inflammatory bowel disease, graft-versus-host disease, malaria, mycobacterial infection, meningitis, psoriasis, congestive heart failure, fibrotic diseases, cachexia, graft rejection, cancers such as cutaneous T-cell lymphoma, diseases involving angiogenesis, autoimmune diseases, skin inflammatory diseases, inflammatory diseases of the eye, inflammatory bowel diseases such as Crohn's disease and colitis, osteo- and rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, adult Still's disease, uveitis, Wegener's granulomatosis, Behcet disease, Sjoegren's syndrome, sarcoidosis, conjunctivitis, polymyositis, dermatomyositis, multiple s
- bacterial or microbial infections comprise any infection with gram positive or gram negative bacteria.
- Pathological conditions of microbial infections include, but are not limited to, conditions such as chronic upper respiratory disease, wound infection, osteomyelitis, endocarditis, skin polymicrobial infections, bronchial asthma, chronic sinusitis, cystic fibrosis or acne vulgaris.
- fungal infections include for example blastomycosis, coccidiodomycosis, cryptococcosis, histoplasmosis, sporotrichosis, chromoblastomycosis, lobomycosis, dermatophytosis, derniatomycosis, onychomycosis, piedra, mycetoma, fusariosis, pityriasis versicolor, tinea barbae, tinea capitis, tinea corporis, tinea cruris, tinea favosa, tinea nigra, tinea pedis, phaeohyphomycosis, rhinosporidiosis, aspergillosis, mycotic keratitis, candidiasis.
- Viral infections to be treated may be caused by viruses including, but not limited to, lentiviruses such as human immunodeficiency virus types 1 and 2 (HIV), human T-cell lymphotropic virus type 1 and 2 (HTLV-I and HTLV-II), SIV, EIAV (equine infectious anemia virus), BIV, FIV, CAEV, VMV, and MMLV (Moloney murine leukemia virus).
- viruses including, but not limited to, lentiviruses such as human immunodeficiency virus types 1 and 2 (HIV), human T-cell lymphotropic virus type 1 and 2 (HTLV-I and HTLV-II), SIV, EIAV (equine infectious anemia virus), BIV, FIV, CAEV, VMV, and MMLV (Moloney murine leukemia virus).
- HIV human immunodeficiency virus types 1 and 2
- HTLV-I and HTLV-II human T-cell lymphotropic virus type 1 and 2
- Such viral infections can also be caused by hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis E virus, hepatitis G virus, human foamy virus, or by human herpes viruses (e.g., herpes simplex virus type-1, herpes simplex virus type-2, herpes simplex virus type-3 (also known as Varicella-zoster virus), herpes simplex virus type-4 (also known as Epstein Barr virus or EBV), herpes simplex virus type-5, herpes simplex virus type-7).
- herpes simplex virus type-1 herpes simplex virus type-2, herpes simplex virus type-3 (also known as Varicella-zoster virus), herpes simplex virus type-4 (also known as Epstein Barr virus or EBV), herpes simplex virus type-5, herpes simplex virus type-7).
- herpes simplex virus type-1 e.g., herpes simplex virus type-1,
- Such viral infections can also be caused by influenza viruses (types A, B or C), human parainfluenza viruses, respiratory syncytial virus, smallpox virus (variola virus), monkeypox virus, vaccinia virus, human papilloma virus, human parechovirus 2, mumps virus, Measles virus, Rubella virus, Semliki Forest virus, West Nile virus, Colorado tick fever virus, foot-and-mouth disease virus, Ebola virus, Marburg virus, polyomavirus, TT virus, Lassa virus, lymphocytic choriomeningitis virus, vesicular stomatitis virus, rotavirus, varicella virus, parvovirus, cytomegalovirus, encephalitis viruses, adenovirus, echovirus, rhinoviruses, filoviruses, coxachievirus, coronavirus (such as SARS-associated coronavirus), Dengue viruses, yellow fever virus, hantaviruses, regional hemorrhagi
- parasitic infections include but are not limited to malaria.
- Parastic infections may be caused by organisms such as protozoa, helminths, and ectoparasites.
- selenium is added as a micronutrient to the composition comprising Aronia extract.
- Selenium is added in one or more suitable form(s) as set forth above.
- selenium is added to the composition to a final concentration from about 0.0001 ⁇ g/mg to about 2.0 ⁇ g/mg.
- selenium may be added to the composition in a range from about 0.0005 ⁇ g/mg to about 1.0 ⁇ g/mg. Even more preferred from about 0.001 ⁇ g/mg to about 0.5 ⁇ g/mg. Still more preferred is a concentration ranging from about 0.001 ⁇ g/mg to about 0.25 ⁇ g/mg. Even still more preferred is a concentration ranging from about 0.001 ⁇ g/mg to about 0.2 ⁇ g/mg. Most preferred is a concentration between about 0.0012 ⁇ g/mg to about 0.1 ⁇ g/mg.
- a fluid composition of the present invention comprises selenium in a concentration from about 0.0001 ⁇ g/mg to about 0.01 ⁇ g/mg, more preferably from about 0.0005 ⁇ g/mg to about 0.005 ⁇ g/mg, most preferably from about 0.0008 ⁇ g/mg to about 0.003 ⁇ g/mg and even most preferably about 0.0012 ⁇ g/mg.
- a solid or dried composition of the present invention comprises selenium in a concentration from about 0.001 ⁇ g/mg to about 0.5 ⁇ g/mg, more preferably from about 0.005 ⁇ g/mg to about 0.01 ⁇ g/mg, most preferably from about 0.003 ⁇ g/mg to about 0.02 ⁇ g/mg and even most preferably about 0.01 ⁇ g/mg.
- the values given are intended to mean the final concentration of added selenium in the composition according to the present invention, i.e. the concentration with which the added selenium is administered to a patient or a subject in need thereof.
- the final concentration of selenium added to the composition according to the present invention may be chosen also in view of the daily requirement amounts of selenium. Accordingly, factors like for example the intended way of administration, and the amount/dosage of the composition given per day and subject, may be considered. Such considerations are within the general knowledge of the skilled person.
- zinc may be added to the composition comprising Aronia extract and selenium, in one or more suitable forms as set forth above, to formulate a composition according to the present invention.
- Zinc may be added to the composition of the present invention in a concentration from about 0.0001 mg/mg to about 0.5 mg/mg.
- zinc may be added to the composition in a range from about 0.0005 mg/mg to about 0.1 mg/mg. Even more preferred from about 0.001 mg/mg to about 0.08 mg/mg. Still more preferred is a concentration ranging from about 0.0015 mg/mg to about 0.006 mg/mg. Even still more preferred is a concentration ranging from about 0.002 mg/mg to about 0.04 mg/mg.
- the values given are intended to mean the final concentration of the added zinc in the composition according to the present invention, i.e. the concentration with which the added zinc is administered to a patient or a subject in need thereof.
- the final concentration of zinc added to the composition according to the present invention may be selected also in view of the daily requirement amounts of zinc. Accordingly, factors like for example the intended way of administration and the amount/dosage of the composition given per day and subject, may be considered. Such considerations are within the general knowledge of the skilled person.
- each of the micronutrients may be added in a concentration or a concentration range as given above
- composition according to the present invention may be administered by itself, for example in liquid or in solid form, or may be formulated as pharmaceutical composition or as nutraceutical composition.
- Said pharmaceutical composition or nutraceutical composition comprises as an active ingredient a composition according to the present invention and at least one pharmaceutically/ nutraceutically acceptable excipient or a mixture of pharmaceutically/nutraceutically acceptable excipients.
- Suitable excipients include, but are not limited to diluents, carriers, disintegrants, binders, glidants, lubricants, coating agents and the like or mixtures thereof, which might be needed for the preparation of final dosage forms.
- the pharmaceutical or nutraceutical compositions according to the present invention may be prepared by mixing the active ingredient, optionally in combination with other active compounds, with one or more suitable excipient(s) by methods which are well known to the skilled person.
- the pharmaceutical or nutraceutical compositions are prepared under sterile conditions.
- the pharmaceutical composition or nutraceutical composition according to the present invention may comprise from about 1% to about 99% by weight of said pharmaceutically acceptable excipient(s), preferably from about 10% to about 95% by weight, and most preferably from about 20% to about 75% by weight.
- excipient(s) The selection of one or more appropriate excipient(s) and their respective amounts is within the general knowledge of the skilled person.
- the excipient(s) may be selected with regard to the intended route of administration.
- the pharmaceutical or nutraceutical compositions according to the present invention may be formulated in a variety of ways to be administered to humans and/or animals.
- the pharmaceutical or nutraceutical composition according to the present invention may be formulated in a liquid form, i.e. for example in the form of solutions, dispersions, emulsions and gels, or in a solid form.
- the pharmaceutical composition as well as the nutraceutical composition may be designed for immediate release and/or for sustained release. Accordingly, also depot forms of the composition according to the present invention are within the scope of the present invention.
- the composition is formulated as a pharmaceutical composition.
- Said pharmaceutical composition comprises a therapeutically effective amount of a composition according to the present invention together with one or more pharmaceutically acceptable excipient(s).
- a therapeutically effective amount of a composition according to the present invention is an amount that when administered to a patient or a subject in need thereof is capable of exerting an immune stimulating effect. In particular, said effective amount is capable of increasing the macrophage activity.
- a pharmaceutical composition according to the present invention may be formulated for [c1] oral, such as peroral, buccal, sublingual, or mucosal administration, or for topical administration.
- said pharmaceutical composition is designed for oral administration.
- a pharmaceutical composition may be designed as an ingestable solution or suspension, comprising the active ingredient in dissolved or suspended form, or as a solid administration form, such as for example tablets, capsules, dragees, powders, granules.
- said pharmaceutical composition is designed for topical administration.
- Topical administration may be achieved for example, but not limiting the present invention thereto, in the form of ointments, creams, lotions, solutions, and elixirs.
- composition according to the present invention since the action of the composition according to the present invention is based on a stimulation of the individual's own immune system, said composition may well be used for the preparation of a nutraceutical composition designed for stimulating the immune system, in particular by increasing the macrophage activity. Therefore, and in accordance with another preferred embodiment of the present invention the composition is formulated as a nutraceutical composition.
- nutraceutical composition is intended to mean a composition that may be considered as food or a food supplement, which is used to improve the diet in order to provide well-being, health benefits and to help to prevent diseases and infections by stimulating the individual's own immune system.
- a pharmaceutical/ nutraceutical composition according to the present invention comprises an effective amount of a composition according to the present invention together with one or more pharmaceutically/ nutraceutically acceptable excipient(s).
- An effective amount of a composition according to the present invention is an amount that when administered to a subject in need thereof is capable of exerting an immune stimulating effect.
- nutraceutical composition according to the present invention may be formulated for oral, such as peroral, buccal, sublingual, or mucosal administration.
- nutraceutical composition is designed for oral administration.
- a nutraceutical composition for oral administration may be designed as an ingestable solution or suspension, comprising the active ingredient in dissolved or suspended form, or as a solid administration form, such as in the form of tablets, capsules, dragees.
- the nutraceutical composition according to the present invention may be formulated to be added as a food supplement to daily food products.
- the nutraceutical composition may be added to beverages, cereals, etc.
- the amount to be administered depends on the subject to be treated taking into account its age, weight and other personal conditions.
- Fresh-frozen Aronia fruits have been used for preparing the Aronia extract (botanical species: Aronia melanocarpa ).
- the aqueous pressed juice was concentrated to obtain a ratio of fruits to pressed juice concentrate (native) of 8-10 to 1.
- Cells Cell line (RAW 264.7, murine macrophages) was purchased from the America Type Culture Collection and cultivated as recommended.
- HBSS Hank's Balanced Salt Solution
- HBSS Hank's Balanced Salt Solution
- MEM minimal essential medium
- the reaction mixture consisted of a suspension (0.25 ml) at a concentration of 5 ⁇ 10 6 PMNL (polymorphonuclear leucocytes)/ml respectively monocytes/macrophages, 50 ⁇ l zymosan, and 0.45 ml medium containing 2 ⁇ 10 ⁇ 6 M lucigenine.
- the reaction mixture was kept at 37° C. before chemoluminescence measurements were performed with a Luminometer (Luminoskan 1251, Labsystems) according to manufacturer's instructions (User's Manual 1995).
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Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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EP09006414.8 | 2009-05-12 | ||
EP09006414A EP2260856A1 (fr) | 2009-05-12 | 2009-05-12 | Composition de stimulation immunitaire comportant un extrait d'arona s en combinaison avec du sélénium et/ou du zinc |
PCT/EP2010/056190 WO2010130629A1 (fr) | 2009-05-12 | 2010-05-06 | Composition de stimulation immunitaire comprenant un extrait d'aronia sp. en combinaison avec du sélénium et/ou du zinc |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/EP2010/056190 A-371-Of-International WO2010130629A1 (fr) | 2009-05-12 | 2010-05-06 | Composition de stimulation immunitaire comprenant un extrait d'aronia sp. en combinaison avec du sélénium et/ou du zinc |
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US14/600,837 Division US10293014B2 (en) | 2009-05-12 | 2015-01-20 | Immune stimulating composition comprising an extract of Aronia sp. in combination with selenium and/or zinc |
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US20120148683A1 true US20120148683A1 (en) | 2012-06-14 |
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US13/320,021 Abandoned US20120148683A1 (en) | 2009-05-12 | 2010-05-06 | Immune stimulating composition comprising an extract of aronia sp. in combination with selenium and/or zinc |
US14/600,837 Active 2030-11-15 US10293014B2 (en) | 2009-05-12 | 2015-01-20 | Immune stimulating composition comprising an extract of Aronia sp. in combination with selenium and/or zinc |
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US14/600,837 Active 2030-11-15 US10293014B2 (en) | 2009-05-12 | 2015-01-20 | Immune stimulating composition comprising an extract of Aronia sp. in combination with selenium and/or zinc |
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US (2) | US20120148683A1 (fr) |
EP (2) | EP2260856A1 (fr) |
JP (1) | JP5612670B2 (fr) |
CN (2) | CN102421446A (fr) |
CA (1) | CA2757759C (fr) |
CY (1) | CY1121268T1 (fr) |
DK (1) | DK2429560T3 (fr) |
ES (1) | ES2704025T3 (fr) |
LT (1) | LT2429560T (fr) |
MX (1) | MX347299B (fr) |
PL (1) | PL2429560T3 (fr) |
PT (1) | PT2429560T (fr) |
TR (1) | TR201818784T4 (fr) |
WO (1) | WO2010130629A1 (fr) |
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CN116270822A (zh) * | 2023-04-21 | 2023-06-23 | 莱檬(清远市)生物科技有限公司 | 一种缓解肺部纤维化的植物提取物及其制备方法和应用 |
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JP2013138633A (ja) * | 2011-12-28 | 2013-07-18 | Pola Chemical Industries Inc | 液状食品組成物の澱発生防止方法 |
CN103417668A (zh) * | 2013-07-29 | 2013-12-04 | 辽宁大学 | 西伯利亚花楸果实提取物在制备抗HeLa癌细胞的药品或保健品中的应用 |
CN105434465B (zh) * | 2015-12-14 | 2019-07-16 | 米军 | 一种防治关节炎的药物组合物及其制备方法 |
JP6961100B2 (ja) * | 2017-10-27 | 2021-11-05 | ジェイビーケーラブ カンパニー リミテッドJbklab Co., Ltd. | アントシアニン−フコイダン複合体を有効成分として含有する免疫増強剤、免疫抗ガン剤及び抗ガン剤の副作用緩和剤 |
US10568866B1 (en) * | 2018-10-05 | 2020-02-25 | Matthias W. Rath | Composition and its use for increasing innate immune health |
CN109248202A (zh) * | 2018-11-14 | 2019-01-22 | 吉林大学 | 一种花楸果及其提取物在制备抗病毒性肝炎药物中的应用 |
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CN116270822A (zh) * | 2023-04-21 | 2023-06-23 | 莱檬(清远市)生物科技有限公司 | 一种缓解肺部纤维化的植物提取物及其制备方法和应用 |
Also Published As
Publication number | Publication date |
---|---|
PT2429560T (pt) | 2019-02-01 |
EP2260856A1 (fr) | 2010-12-15 |
MX347299B (es) | 2017-04-21 |
CA2757759A1 (fr) | 2010-11-18 |
JP2012526083A (ja) | 2012-10-25 |
TR201818784T4 (tr) | 2019-01-21 |
PL2429560T3 (pl) | 2019-04-30 |
CY1121268T1 (el) | 2020-05-29 |
CA2757759C (fr) | 2016-12-20 |
WO2010130629A1 (fr) | 2010-11-18 |
MX2011011860A (es) | 2011-12-08 |
ES2704025T3 (es) | 2019-03-13 |
CN103845449A (zh) | 2014-06-11 |
US10293014B2 (en) | 2019-05-21 |
US20150132418A1 (en) | 2015-05-14 |
JP5612670B2 (ja) | 2014-10-22 |
CN102421446A (zh) | 2012-04-18 |
LT2429560T (lt) | 2018-12-10 |
DK2429560T3 (en) | 2019-02-18 |
EP2429560B1 (fr) | 2018-11-14 |
EP2429560A1 (fr) | 2012-03-21 |
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