cu») United States ‘US 2010009258541 «2 Patent Application Publication co) Pub. No.: US 2010/0092585 Al oy ra) 3) ey @ PLANT EXTRACTION METHOD AND COMPOSITIONS hers, Tertell, TX CComespondence Address ‘S OLSON & BEAR LLP IRVINE, CA 92614 (US) Antonio, TX (US) (43) Pub, Date Apr. 15, 2010 Related US. Application Data (60) Provisional pplication No, 61/105,133. fled on Oct. 14,2008, ication Classification G1) Ine AGIK 36/886 (2006.01) 4610 19/00 (2006.01), cO7G 3700 (2006.01), (2) US.CL 24TH; 53618.5 on ABSTRACT The present invention pertains to methods of extracting cr diac’ glycosides fom cardiac glyeoside containing plant ‘material, such as Nerium oleander, through use of ale. It Further provides for compositions resuling Irom such extrac- ‘ions, pharmaceutical compositions, cosmetie compositions, ‘and methods of eating skin conditions,US 2010/0092585 AI PLANT EXTRACTION METHOD AND. COMPOSITIONS, (CROSS-REFERENCE TO RELATED "APPLICATIONS, {0001} This application claims the benefit of U.S. Provi sional Application No, 61/105,133, fled on Oct. 14, 2008, ‘hich is incorporated herein by reference in its entity. FIELD OF THE INVENTION [0002] The present invention pertins 10 methods of ‘extracting materials from biological sources and particularly from plant mater for cosmetic and medicinal applications DESCRIPTION OF THE RELATED ART 10003] Biological sources have provided the bases for medicines and cosmetics Irom the cariest days of mankind. Most suc sources have been plants, which initially were used sis. Oftentimes, however, iis desirable to extract materials, rom plans, as for example when the desired material con- stitutes only a small proportion ofa given plant, or when the ‘material ceurs in he presence of other, undesirable constiti- [10003] | Giycosides represent an important class of com pounds extricted from plant sources. Cardiac glycosides, for ‘example, are alycosylated steroids, i, steroids conjugated to a carbohydrate moiety. Examples of eardive glycosides include those useful in the production of such drags as ‘digoxin and digiloxin, An importat class of glycosides ‘comes from the gems Nerium, which inches such speciesas Nerium indicum, Nerium adorum, an Nerium oleander, the last being th oleander plant native to Asia and the Meslitr- ranean Tittoral and now found also in parts of the United States. The various Nerium species aor such glycosides as ‘leanda, which finds extensive use in medicine, 10005] Extraction of cardiac glyoosides from Nerium ole- ander filitates their use clinically by allowing administra tion of compasitions of known potency and purity. Previous methods of extracting these glycosides have employed hot Water, a deseribed in U.S, Pat, Nos, 135,745 and 6,565, 807, as well as ULS. Patent Publication No. 20060188585, ‘each of which is hereby incosporated By reference in is ‘entirety, Owing to the lage lipophilic steroidal moiety. how= ‘ever, oleandrin has poor agus solubility, which limits the ‘effectiveness of aquoous extraction. Furthermore, plants gen- ‘erally have waxy cuticles surrounding thee exterior surfaces to minimize desiccation, This waxy cuticle not only limits the ‘epress of water Irom the plant fo the exterior, but also the Ingress of water from the exterior into the plant, and thus Jurther impedes efficient aqueous extraction of the desired cardiac glycosides. Iso, the desired cardiac glycosides can hydrolyze or otherwise decompose on long exposure fo hot 10006] One solution to these problems has been to perfon theextaetion ol with pure water but witha mixtureof Water anda water-miseiblealeohol, such as methanol or ethanol, as described in US. Patent Publication No,, 20070154573, ‘hich is hereby incorporated by reference in is entirety. Use ‘of aqueous afoahol increases both the peneteation of the ‘extraction solvent into the interior of the plant and the sol bility ofthe cardiac glycosides to some extent. More rcealy, US. Pat. No. 7,402,325, which is hereby incorporated by reference in its entirety, has described use of supercritical Apr. 15, 2010 CO, as extracting higher yields of desired product from pow dered oleander leaves. Exinction with supercritical CO, necessitates use of high pressure apparatus (above ea. 100 ‘tmospheres), with all ofits attendant hazard, [0007] Accordingly. need exists for a way to extract ole- Aandgin and other eardiae glycosides more efficiently from ‘various plant species such ts Nerium without use of excessive heat or high pressure apparatus SUMMARY OF THE INVENTION [0008] ‘Tae present invention provides a method to extract cardiac ulyeosides from a cardia plyeoside-containing pl species. sichats a Nerium species, throughuse of aloe sich as {hat derived fom lloe ver. Ic Turther provides composito resulting from such extraction. [0009] "Specifically. it provides a method of performing an extraction of plant material derived from acardiac glycoside- containing plant species, such as a species from the penus Nerium, comprising agitating the plant materia in aloe Jage and separating the extract from any remaining solid material 0010] Tae extraction method optionally involves heating the solution from about 40° C. to aboot 100° C., optionally including use of extraction adjuvants such a alcobols, Ketones, and esters [0011] "An embodiment provides a method of extracting cardiac glycosides comprising intermixing a cardiac glyeo- ‘ce plant species with aloe uncer conditions selected a orm Aan extraction mixture 0012] An embodiment further provides conditioning the exttacion mixture under conditions selected to extract cat tac glycosides from the cardiae glycoside plant species to {orm a eondiioned extraction mixture, wherein the condi- tioned extriction mixture comprises rexidual canlixe lyco- side plant species and a candae glycoside aloe mixture [0013] An embodiment further provides separating at least 4 potion of the cardia glycoside aloe mixture from the residual caniac glycoside plant species 10 form a cardiac alyooside aloe extract, where the eardioe glycoside aloe extract comprises cardiae glycosides extracted froma carding slyonside plant species, and where the eariae glycoside aloe extract is substantially free of the residual cardia glycoside plant species [014] In some embodiments the cardiac glycoside plant species helongs toa family elected from Apocynaceae, Bras- ‘escene, Plantaginaceae, Ruscaceae, of Hyacinthsceze. [015] In some embodiments the cardiae glycoside plant belongs to the species Neriume indicuim or Nerium oleander [0016] In some embodiments the conditioning comprises beating theextmetion mixtoreto a tempernturein the rangeot ahout 40° C.to about 100°C. o form the conditioned exime- [0017] In some embodiments the conditioning. comprises ‘eating the extraction mixture fora heating ime inthe range ff about Io about 10 hours. [018] In some embodiments the separating comprises subjecting the conditioned extaction mixture to a separation pethod selected from the group consisting o ilration, cen tefiagation, and decanting [0019] In some embodiments the extraction mixture com- prises an amount of the eandiae glycoside plant species inthe range of about one pat to about 50 parts by weight and aaUS 2010/0092585 AI tne range of about one pat to about 100 jon the toll weight of extrction mix 10020) Prisesan adjuvant sclected from the group consisting of aleo- hols, Ketones and esters, In some embodiments the extraction mixture com= 10021] An embodiment provides a caniae glycoside aloe ‘composition, comprising aloe and at least one eardiae glyco- side, 10022] In some embodiments, the cardiac glycoside is @ ‘cardiac alycoside aloe extract from a cardiac glycoside plant species. 10023] In some embodiments, the canliae glycoside aloe ‘composition is eardiae glycoside aloe extract wherein the ‘cardiac slycoside plant species is Nerium oleander 0024) "An embodiment provides pharmaceitical composi tions comprising aloe, atleast one eardiae glycoside and & pharmaceutically acceptable carrier. Another embodiment provides cosmetic compositions comprising aloe, atleast one ‘cardiac slycoside and a demmal agent, 0025) In certain embodiments, a method of treatment is provided comprising identifying subject having askin con- dition and applying an effective amount of the pharmaceut ‘cal composition comprising aloe and at least one cardiac lyeoside to the skin of a subject to thereby treat the skin condition, 10026] In certain embodiments the skin condition is Sclecte from the group consisting of abscesses, acne, actinic Keratoss, age spots, liver spots, burns, sunburn, heat burn, radiation bur, cold sores, coms, eczema, psoriasis, ring” ‘wom, scabies, skin cancers, basal skin cancer, squamous skin cancer, melanoma skin Cancer, skin tags, and wars. [0027] These and other embodiments are described in rear detail below DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 10028] An embodiment provides a method of extracting ‘cardiac glycosides, comprising internixing a cardiac glyco- side plant species with aoe under conditions selected 0 form ‘an extraction mixtre 10029] Examples ofcandiae glycoside plant species include those inthe family Apoeynaceae (dogbane), particularly in the genera Nerium, Sirophanthus, Apocymum, Thevetia, and Catharanthus, the family Brassicaceae, particularly in the enus Chemanthus, the family Plantaginaceae, particularly in the genus Digitalis, the family Ruscacene, particularly inthe ens Comvatlaria, ad in the Family Hyacinthacese,particu- iy in he genus Urgine. 10030] | Fxamples of panicular cardse glycoside plant spe- ies inlude Nevium oleander, Thevetia nerifolia, Digitalis purpurea, Digitalis lanate, Convallaria majalis, Urginea ‘maritima, Urginea indica, Srophanthus gratus, Apocyrum ‘carnabinum, Chemanths cheir 10031] The erm “aloo” refers toa genus of plants native 10 Aiea and comprising about 400 species, including Aloe arborescens, Aloe arstata, Aloe dickotoma, Aloe nyerienss, Aloe varvegata, Aloe wild, ana Aloe barbadensis miler 10032] In some embodiments, the conditions selected 10 orm an extraction mixture comprise mixing aloe with the leaves and stems ofa cardiae glyeoside plant species that are ‘optionally cot into pices, milled, or powdered facilitate the ‘extraction, Apr. 15, 2010 [0033] In some embodiments, the extraction mixture com- privs an amount of cardise glycoside plant species in the range of about one part to about S0 parts by weight and an ‘amoust of aloe inthe range of about one part to about 100 parts by weight, based on total weight of the extraction mix- ture [0034] An embodiment further provides conditioning the ‘extraction mixture under conditions selected lo extract ca diac glycosides from cardiae glycoside plant species to form ‘conditioned extraction mixture, wherein the conditioned extraction mixture comprisesresidual eardiae glycoside plant species and a cardiac glycoside aloe mixture 10035] Examples of conditioning the extraction mitre inder conditions selected to extract canine glycosides fom cardiac glycoside plant species. includeheating the extraction ‘mixture, agitating the extraction mixture, and heating the extraction mixture with agitation, [0036] In some embodiments, conditioning the extraction mixture under conditions selected to extract cardiae glyeo- Sides from cardiac plyeoside plant species comprises heating the extraction mixture toa temperature in the range of abort 40° C. to about 100° C. to form a conditioned extraction [0037] In some embodiments, conditioning the extraction ture under conditions selected to extract cardiac glyco- sides ftom eardise glycoside plant species comprises heating the extraction mixture for atime in the range of about 1 40 bout 10 hours to form a conditioned extraction mixture 0038} In some embodiments, a conditioned extraction mixture comprises @ mixture where a portion of the cardiae alyoosides from the cardiac glycoside plant species are extracted into the aloe present in the mixture, Thus, the result ‘of conditioning isto form a conditioned extraction mixtire ‘hat contains residual cardiac glycoside plant species and a cardiac glycoside alae mixture that contains both aloe and the cardiae glycosides) extracted into the aloe from the cardiac slyooside plant species. [039] An embodiment further provides separating at least 4 portion of the cardive glyeoside albe mixture from the residual cadiae glycoside plant species 10 form a cardiae alyooside aloe extract. [0040] ‘Those skilled in tho art will appreciate that the cat= ‘ae plyeoside aloe extract may contain, ination to cardi alyeoside(s) other components extracted by thealoe from the cardiac glycoside plant species, such as, for example polysaccharides). [041] Examples of separating at least a portion of the cardiac plyeoside aloe mitre Irom the rsidal eardiae ply coside plant species include filtration, separation, and decant- ing. [0042] In some embodiments, the cardiac glyeoside aloe extract comprises earise glycosides extracted from the ea ‘ac glyeosie plant species. [0043] In some embodiments, the cardne ylyeoside aloe extract is substantially free of the residual cardiae glycoside plant species [0044] In some embodiments, the extraction mixture com- prises an adjuvant sclete! fom the group consisting of aleo- hols, ketones, and esters, [004] _ further embodiment provides a cardia plyeoside aloe composition, comprising aloe and at least ane cari alyeoside,