Patent Application: US-48912309-A

Abstract:
a method of treating a subject requiring immune mediation includes administering to the subject an immunostimulatory composition which contains black raspberry seed oil . administering this composition stimulates the production of certain immune cells , including nk cells and cd8 lymphocytes . administering the composition by this method also provides a method of treating , preventing , or ameliorating a condition or disease in the subject requiring enhanced immune system support .

Description:
for over a decade , we have been investigating methods to treat liver cancer ( adenocarcinoma that metastasized from the colon ) in mice in our laboratory . we postulated that enhancing the mouse &# 39 ; s immune system in part with antioxidants may improve clinical success of the cancer treatment methodology we were researching . certain plant seed oils are extraordinarily high in antioxidants and , for the purpose of our study , black raspberry seed oil was used as a dietary supplement to provide the mice with a source of a variety of antioxidants because it has one of the highest levels of antioxidants . moreover , black raspberry seed oil has a higher oxygen radical - reducing capacity than palm , corn , sunflower or olive oil . the oil was obtained from the black raspberry seeds via the cold press manufacturing method . the cold press extraction method yields excellent quality oil with minimal chemical decomposition compared to traditional high temperature extraction methods . when using cold - pressing , nothing is added to the seeds or to the oil to increase production during the manufacturing process . the oils are pure and unadulterated . in the study , black raspberry seed oil was added daily to the food of the mice and the amount of oil administered was equal to 10 % of the rodent &# 39 ; s weight . after varying periods of time , the mice were euthanized and their body chemistry carefully examined . in the course of this research , it was discovered that the highly potent antioxidant oil , black raspberry seed oil , appeared to augment the populations of both nk ( natural killer cells ) and cd8 (+) t ( cytotoxic t lymphocyte ) cells . this research is key because an increase in the number of nk and cd8 (+) cells enables to body to better fight the damage caused by free radicals and enhance the body &# 39 ; s ability to fight disease and cancer . evaluation of splenic lymphocyte populations when black raspberry seed oil is administered mice were sacrificed for spleen harvest and to obtain cardiac blood samples . spleens were processed for flowcore analysis and the data processed using statview . cardiac blood was centrifuged to collect serum for elisa analysis of circulating cytokines at a later date . recent studies have shown that plant oils rich in antioxidants also increase cellular immune response . we postulated that the addition of antioxidant oil to a murine diet hypothesis , animals were randomly placed into two groups : diet with and without oil . for the duration of each study , the animals were fed standard rodent chow or standard rodent chow supplemented with 10 % ( by weight ) black raspberry seed oil , a potent antioxidant plant oil . splenic lymphocyte populations were analyzed 3 , 7 , and 14 days . we found significantly elevated cd8 (+) lymphocyte populations in the oil group at all time points when compared to the other group . furthermore , we observed that the nk population peaked on day 7 . these results suggest that antioxidant oil in conjunction with the normal diet augments cytotoxic immune response . fig1 shows the effect of black raspberry seed oil diet on natural killer ( nk ) cell populations . after splenocytes were isolated , flow cytometry was utilized to determine the percentage of nk cells present . each value represents the mean + sd of at least 5 mice . error bars are not apparent if less than 0 . 5 . fig2 shows the effect of black raspberry seed oil diet on cytotoxic t lymphocyte ( cd8 +) populations . after splenocytes were isolated , flow cytometry was utilized to determine the percentage of cd8 (+) lymphocytes present . each value represents the mean ± sd of at least 5 mice . error bars are not apparent if less than 0 . 5 . in experiments performed at the university of minnesota medical school , black raspberry seed oil was used in conjunction with a form of salmonella that does not cause disease and which has been genetically altered to carry a gene for a powerful immune stimulant . it is known that salmonella readily invades tumor tissue and we have successfully used the organism to deliver the immune stimulant . combining the salmonella with black raspberry oil further enhances our immune stimulation and tumor destruction in mice with liver cancers . fig3 is an example of an experiment showing tumor reduction in a two week period using the potent antioxidant oil alone and in combination with the salmonella carry a gene for immune stimulant . since the present seed oil preparations maybe useful as agents for immunotherapy in the treatment of immunodeficiency disorders , cancer , wound healing and infectious diseases , the present invention includes pharmaceutical compositions containing the instant seed oil preparations optionally in combination with acceptable pharmaceutical carriers or excipients . pharmaceutical compositions suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve its intended purpose . more specifically , a therapeutically effective amount means an amount effective to prevent development of or to alleviate the existing symptoms of the subject being treated . determination of the effective amounts is well within the capability of those skilled in the art , especially in light of the detailed disclosure provided herein . the amount of composition administered will be dependent upon the condition being treated , the subject being treated , on the subject &# 39 ; s weight , the severity of the affliction , the manner of administration and the judgment of the personalizing physician . the pharmaceutical compositions of the present invention may be manufactured in a manner that is itself known , e . g ., by means of conventional mixing , dissolving , granulating , dragee - making , levigating , emulsifying , encapsulating , entrapping or lyophilizing processes . pharmaceutical compositions for use in accordance with the present invention thus may be formulated in conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the compositions compounds into preparation which can be used pharmaceutically . proper formulation is dependent upon the route of administration chosen . for injection , the agents of the invention may be formulated in aqueous solutions , preferably in physiologically compatible buffers such as hanks solution , ringer &# 39 ; s solution , or physiological saline buffer . for transmucosal administration , penetrants appropriate to the barrier to be permeated are used in the formulation . such penetrants are generally known in the art . for oral administration , the compositions can be formulated readily by combining the active compositions with pharmaceutically acceptable carriers well known in the art . such carriers enable the compounds of the invention to be formulated as tablets , pills , dragees , capsules , liquids , gels , syrups , slurries , suspensions and the like , for oral ingestion by a patient to be treated . pharmaceutical preparations for oral use can be obtained as a solid excipient , optionally grinding a resulting mixture , and processing the mixture of granules , after adding suitable auxiliaries , if desired , to obtain tablets or dragee cores . suitable excipients are , in particular , fillers such as sugars , including lactose , sucrose , mannitol , or sorbitol ; cellulose preparations such as , for example , maize starch , wheat starch , rice starch , potato starch , gelatin , gum tragacanth , methyl cellulose , hydroxypropylmethyl - cellulose , sodium carboxymethyl cellulose , and / or polyvinylpyrrolidone ( pvp ). if desired , disintegrating agents may be added , such as the cross - linked polyvinyl pyrrolidone , agar , or alginic acid or a salt thereof such as sodium alginate . dragee cores are provided with suitable coatings . for this purpose , concentrated sugar solutions may be used , which may optionally contain gum arabic , talc , polyvinyl pyrrolidone , carbopol gel , polyethylene glycol , and / or titanium dioxide , lacquer solutions , and suitable organic solvents or solvent mixtures . dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses . pharmaceutical preparations which can be used orally include push - fit capsules made of gelatin , as well as fit , sealed capsules made of gelatin and a plasticizer , such as glycerol or sorbitol . the push - fit capsules can contain the active ingredients in admixture with filler such as lactose , binders such as starches , and / or lubricants such as talc or magnesium stearate and , optionally , stabilizers . in soft capsules , the active compounds may be dissolved or suspended in suitable liquids , such as fatty oils , liquid paraffin , or liquid polyethylene glycols . in addition , stabilizers may be added . all formulations for oral administration should be in dosages suitable for such administration . for buccal administration , the compositions may take the form of tablets or lozenges formulated in conventional manner . for administration by inhalation , the compositions for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebulizer , with the use of a suitable propellant , e . g ., dichlorodifluoromethane , trichlorofluoromethane , dichlorotetrafluoroethane , carbon dioxide or other suitable gas . in the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount . capsules and cartridges of e . g ., gelatin for use in an inhaler or insufflator may be formulated containing a power mix of the compound and a suitable powder base such as lactose or starch . the compositions may be formulated for parenteral administration by injection , e . g ., by bolus injection or continuous infusion . formulations for injection may be presented in unit dosage form , e . g ., in ampoules or in multidose containers , with an added preservative . the compositions may take such forms as suspensions , solutions or emulsions in oily or aqueous vehicles , and may contain formulatory agents such as suspending , stabilizing and / or dispersing agents . pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water - soluble form . additionally , suspensions of the active composition may be prepared as appropriate oily injection suspensions . suitable lipophilic solvents or vehicles include fatty oils such as sesame oil , or synthetic fatty acid esters , such as ethyl oleate or triglycerides , or liposomes . aqueous injection suspensions may contain substances which increase the viscosity of the suspension , such as sodium carboxymethyl cellulose , sorbitol , or dextran . optionally , the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions . alternatively , the active ingredient may be in powder form for constitution with a suitable vehicle , e . g ., sterile pyrogen - free water , before use . the compositions may also be formulated in rectal compositions such as suppositories or retention enemas , e . g ., containing conventional suppository bases such as cocoa butter or other glycerides . in addition to the formulations described previously , the compositions may also be formulated as a depot preparation . such long acting formulations may be administered by implantation ( for example subcutaneously or intramuscularly ) or by intramuscular injection . thus , for example , the compositions may be formulated with suitable polymeric or hydrophobic materials ( for example as an emulsion in an acceptable oil ) or ion exchange resins , or as sparingly soluble derivatives , for example , as a sparingly soluble salt . the pharmaceutical compositions also may comprise suitable solid or gel phase carriers or excipients . examples of such carriers or excipients include but are not limited to calcium carbonate , calcium phosphate , various sugars , starches , cellulose derivatives , gelatin , and polymers such as polyethylene glycols . suitable routes of administration may , for example , include oral , rectal , transmucosal , transdermal , or intestinal administration , parenteral delivery , including intramuscular , subcutaneous , intramedullary injections , as well as intrathecal , direct intraventricular , intravenous , intraperitoneal , intranasal , or intraocular injections . alternatively , one may administer the composition in a local rather than systemic manner , for example , via injection of the compound directly into an affected area , often in a depot or sustained release formulation . furthermore , one may administer the drug in a targeted drug delivery system , for example , in a liposome coated with an antibody specific for affected cells . the liposomes will be targeted to and taken up selectively by the cells . the compositions may , if desired , be presented in a pack or dispenser device which may contain one or more unit dosage forms containing the active ingredient . the pack may for example comprise metal or plastic foil , such as a blister pack . the pack or dispenser device may be accompanied by instructions for administration . compositions comprising a composition of the invention formulated in a compatible pharmaceutical carrier may also be prepared , placed in an appropriate container , and labeled for treatment of an indicated condition . suitable conditions indicated on the label may include treatment of a disease . dietary supplements suitable for use in the present invention include compositions wherein the active ingredients are contained in an effective amount to achieve its intended purpose . more specifically , an effective amount means an amount effective to prevent development of or to alleviate the existing symptoms of the subject being treated . determination of the effective amounts is well within the capability of those skilled in the art , especially in light of the detailed disclosure provided herein . the amount of composition administered will be dependent upon the condition being treated , the subject being treated , on the subjects weight , the severity of the affliction , the manner of administration and the judgment of the personalizing physician . the ingredients of the dietary supplement of this invention are contained in acceptable excipients and / or carriers for oral consumption . the actual form of the carrier , and thus , the dietary supplement itself , may not be critical . the carrier may be a liquid , gel , gelcap , capsule , powder , solid tablet ( coated or non - coated ), tea or the like . suitable excipient and / or carriers include maltodextrin , calcium carbonate , dicalcium phosphate , tricalcium phosphate , microcrystalline cellulose , dextrose , rice flour , magnesium stearate , stearic acid , croscarmellose sodium , sodium starch glycolate , crospovidone , sucrose , vegetable gums , agar , stevia , lactose , methylcellulose , povidone , carboxymethylcellulose , corn starch , and the like ( including mixtures thereof ). the various ingredients and the excipient and / or carrier are mixed and formed into the desired form using conventional techniques . dose levels / unit can be adjusted to provide the recommended levels of ingredients per day in a reasonable number of units . the dietary supplement may also contain optional ingredients including , for example , herbs , vitamins , minerals , enhancers , colorants , sweeteners , flavorants , inert ingredients , and the like . such optional ingredients may be either naturally occurring or concentrated forms . selection of one or several of these ingredients is a matter of formulation , design , consumer preference and end - 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