Note: Descriptions are shown in the official language in which they were submitted.
WO 98/08490101520253035CA 02264718 1999-03-01PCT/U S97/ 15072SOLID COPRECIPITATE8 FOR ENHANCEDBIOAVAILABILITY OF LIPOPHILIC SUBSTANCESFIELD OF THE INVENTIONThe present invention concerns compositions comprisingsolid coprecipitates for improved oral delivery and enhancedbioavailability of lipophilic substances, and to methods forthe preparation and use of these compositions. Moreparticularly, these compositions include tocopherolpolyethyleneglycol succinate powdered coprecipitates oflipophilic substances.BACKGROUND OF THE INVENTIONLipophilic substances possessing low water solubilityoften have poor oral bioavailability. These substances,being hydrophobic by nature, show wetting difficulties andpoor dissolution. These properties obviously represent arate-limiting step in their absorption from solid oral dosageforms and, in turn, cause a subsequent reduction in theirbioavailability.To address the foregoing issue, these lipophilicsubstances are usually administered in the form of liquidpreparations dissolved in edible oils or formulated inoil-inâwater emulsions or microemulsions. Even in theseformulations, however, the oral bioavailability of many ofthem is still very low. Thus,unresolved need for safe and useful formulations that provideeven today, there remains anenhanced oral bioavailability for such substances.Cannabinoids are one example of a family of lipophilicsubstances with very poor water solubility. Cannabinoidssuch as Ahtetrahydrocannabinol (AVTHC),A°âtetrahydrocannabinol (A°âTHC), A9-tetrahydrocannabinolmy-THC), cannabinol, cannabidiol, and their metabolites, arehighly hydrophobic lipid soluble compounds and can bedissolved in aqueous solutions only in the range of a fewmicrograms/ml or less, depending upon the conditions (Garretand Hunt, J. Pharm. Sci., 63:1056-1064,1974).WO 98/08490101520253035CA 02264718 1999-03-01PCT/US97/15072In general, the systemic availability of cannabinoidsof thefollowingafter oral administration is low and mean estimateshuman bioavailability of tetrahydrocannabinol (THC)oral ingestion range from 6 to 12% depending on the vehicleused. For example, the maximal plasma levels after oraldosing of 20 mg THC in a sesame oil formulation were around10 ng/ml (Wall et al., Clin. Ther. 34:352â363,1983).Dexanabinol (also denoted HU-211), is disclosed in USPatents 4,876,276 and 5,521,215, as a synthetic non-psychoactive cannabinoid with novel neuroprotective activityPharmacol.in the multiple-action treatment of brain damage associatedThe chemical(+)-(3S,4S)â7-hydroxywy-tetrahydrocannabinol-1,1âdimetylheptyl, is shown in Scheme 1.onwith stroke, head trauma, and cardiac arrest.structure of dexanabinol, Scheme 1Dexanabinol is a very lipophilic compound which ispractically insoluble in water (less than 50 pg/ml) and, likeother lipophilic drugs, exhibits poor oral bioavailability.Coenzyme Q10, chemically named 2,3âdimethoxy-5-methyl-6-decaprenylâ1,4-benzoquinone and also known by the namesUbiquinone or Vitamin K, is classified as a fat-solubleIt has beenidentified as an essential component of the mitochondrialquinone, a naturally occurring enzyme.respiratory chain, and, thus, a necessary part of a cell'senergy production. More particularly, it constitutes aredox-link between flavoproteins and cytochromes and acts asan electron shuttle controlling the efficiency of oxidative-2-WO 98/08490101520253035CA 02264718 1999-03-01PCT/US97/15072phosphorylation. It is a very lipophilic compound andpractically insoluble in water due to its long side chain of10 isoprenoid units.CoQ10 has been previously identified as a naturalantioxidant with potential use as a dietary supplement toprotect against ageârelated degeneration and as an adjuvantvitamin to prevent or treat many disease states.Supplementary Coenzyme Q10 has reportedly shownbeneficial influences in a variety of conditions or diseases,including: periodontal disease, certain blood circulationdiseases, impaired memory, fatigue, irregular heartbeat, highblood pressure, immune system impairment, and the agingprocess.The recommended daily allowance for coenzyme Q10 has notbeen determined. Most experts agree, however, that the dailyrequirement lies somewhere between 30 and 60 milligrams.When treating illnesses, dosages of 100 to 300 milligrams arecommonly used.The oral bioavailability of CoQ10 is related to thedissolution rate of the formulation (Kishi et a1., Metabolismof exogenous coenzyme Q10 in vivo and the bioavailability ofK. Folkers, and Y.Yamamura, eds., Biomedical and Clinical Aspects of CoenzymeQ10, Vol. 4, 1984,pp. 131-142).Given CoQ10's poor solubility in water, the ability tocoenzyme Q10 preparations in Japan, in:Elsevier, Amsterdam,formulate this substance in a suitable form affordingconvenient and efficient oral bioavailability is a desiredgoal. The art has attempted to address the problem bydisclosing formulations of Coenzyme Q10 using lipids, in theform of emulsions, liposomes, microparticles andnanoparticles, have previously been disclosed. These knownlipid formulations have comprised particles dispersed in anaqueous medium, and are suitable for various routes ofadministration, including primarily intravenousadministration, as disclosed in: WO 95/05164, which disclosesmicroparticles and nanoparticles in aqueous suspension; USPatent 4,824669, which discloses fatty emulsions; US Patent-3.-WO 98/08490101520253035CA 02264718 1999-03-01PC1VUS97ï¬50724,636,381, which discloses liposomes; and US Patent4,483,873, which discloses aqueous dispersions or solutions.The neurohormone melatonin is synthesized in the pinealgland with a nocturnal circadian rhythm. Sleep disorders,seasonal depression, mood disorders, migraine, and jet lagare some of the disorders that have been correlated to adisruption of normal, physiological melatonin secretion.There are reports on the beneficial effect of exogenousmelatonin administration to reestablish the synchronizationof circadian rhythm. However, these studies have shown largevariations in oral melatonin absorption, as well ashighlighted the inconvenience of employing a continuousintravenous delivery system. Therefore, an oral formulationof melatonin with good bioavailability is needed. Amelatonin buccal mucoadhesive sustained release deliverypatch, mimicking endogenous secretion, has already been shownas one alternative means for delivering melatonin (Benes etal., Proceed. Intl. Symp. Control. Rel. Bioact. Mater.,21:551-552, 1994).Additional examples of lipophilic drugs with very poorwater solubility and low oral bioavailability which couldbenefit from oral dosage forms are the peptide drugcyclosporin A, the antifungal agent amphotericin B, theanticancer drug etoposide, as well as tamoxifen and itsanalogs.Water-dispersible vitamin preparations were disclosed inUs Patent 3,102,078, wherein the vitamin E derivativetocopherol polyethyleneglycol succinate ("TPGS") was shown toOilycompositions of anti-tumor drugs utilizing TPGS as ahave useful properties as a solubilizing agent.solubilizing adjuvant have also been disclosed for instancein US 4578391.substance (US 4,668,513), as a cryoprotectant (US 5,198,432)or to improve the bioavailability of vitamin E (US patents5,179,122 and 5,223,268) have also been disclosed.formulation of water dispersible vitamin E compositions forFurther uses of TPGS as a surface activeA powder101520253035CA 02264718 1999-03-01PCTIUS97/15072[pm/us01sEP1998use as a vitamin E supplement has also been disclosed in US5,234,595.The structure of TPGS is shown in scheme 2atHâ mwahah=amaa0%coo.L., on.L.Lmï¬wï¬ï¬ï¬ï¬iScheme 2where n represents the average number of âCH§HgO- groups inthe polyethylene glycol chain. For typical polyethyleneglycols, n is from about 5.2 (PEG 200) to about 182.4 (PEG8000). Thus, there remains a need for new formulations whichprovide improved oral bioavailability of such compounds. Thepresent invention provides such formulations.SUMMARY OF THE INVENTIONThis invention is directed to compositions comprisingtocopherol polyethyleneglycol succinate (TPGS) solidcoprecipitates useful for the oral delivery of lipophilicsubstances with low oral bioavailability, and to methods forpreparing and using such compositions.The solid coprecipitate compositions of the presentinvention include a lipophilic substance in an amountsufficient to provide a therapeutic effect when administeredto a mammal; tocopherol polyethyleneglycol succinate (TPGS)in an amount sufficient to increase the oral availability ofthe lipophilic substance; and a dispersion adjuvant in anamount sufficient to assist in dispersing the lipophilicsubstance in the succinate.According to a preferred embodiment of the presentinvention, the solid TPGS coprecipitates of the presentinvention comprise a dispersion adjuvant selected from the-5-AMENDED SHEETCA 02264718 1999-03-01PC!/U397/15072lP5Ws01sEP1998group consisting of polyvinylpyrrolidone, a medium chaintriglyceride, a long chain triglyceride, tocopherol acetate,and polyethyleneglycol...5A-AMENDED SHEâWO 98/08490101520253035CA 02264718 1999-03-01PCT/US97l1 5072According to a more preferred embodiment, the solid TPGScoprecipitates of the present invention, comprisepolyvinylpyrrolidone ("PVP") as the dispersion adjuvant.According to a most preferred embodiment of the presentinvention, the solid TPGS coprecipitates of the presentinvention may advantageously further comprise a known free-Thisembodiment provides powdered TPGS coprecipitates particularlyflow imparting agent, such as fumed silica or the like.useful in the preparation of solid dosage forms for oraladministration.Lipophilic substances incorporated in the powdered TPGScoprecipitates of the present invention have shownunexpectedly improved drug release in simulated gastric fluidin vitro and enhanced oral bioavailability in vivo.The present invention further relates to methods forproducing the powdered TPGS coprecipitate compositions,comprising:co-melting TPGS and the lipophilic substance at 40â60°C;adding a dispersion adjuvant to the melted mixture withagitation;adding a fumed silica to the mixture with agitation; anddrying the resultant mixture at 100°C to get a drycoprecipitate.The dispersion adjuvant used in accordance with thismethod may be added as an aqueous solution, an organiccosolvent solution, or an oil.According to yet another embodiment, the solid TPGScoprecipitate compositions according to the present inventionmay be prepared by freeze-drying the TPGS/lipophilicsubstance/dispersing adjuvant mixture.According to yet another embodiment, the solid TPGScoprecipitates compositions according to the presentinvention may be prepared by sprayâdrying the TPGS/lipophilic substance/dispersing adjuvant mixture.According to a more preferred embodiment, the TPGS/lipophilic substance/dispersing adjuvant coprecipitateformulations can be spray dried or freeze-dried to obtain dry-6-101520253035CA 02264718 2005-01-11powdered compositions, suitable for the preparation ofsolid-dosage forms such as hard gelatin capsules or tablets.According to a still more preferred embodiment, thesesolid dosage TPGS coprecipitate compositions are advantageousfor the oral delivery of Coenzyme Q10 as a dietary nutrientsupplement, melatonin, dexanabinol, amphotericin B,cyclosporin A, etoposide, tamoxifen quaternary amine analogs,or for any appropriate lipophilic substance.BRIEF DESCRIPTION OF THE DRAWINGSFIG. 1 shows the in vitro release patterns ofDexanabinol in simulated gastric fluid from powdered TPGS/PVPsolid lipid coprecipitate compositions containing about 2.5mg drug prepared as described in Example 1 (Table 1) andpacked in hard gelatin capsules.FIG. 2 shows the in vitro release patterns ofDexanabinol in simulated gastric fluid from powderedTPGS/PVP/Miglyolâ solid lipid coprecipitate compositionscontaining about 2.5 mg drug prepared as described in Example1 (Table 2) and packed in hard gelatin capsules.FIG.oral administration (50 mg/kg drug, n=5) of a solid TPGS3 shows the Dexanabinol rat plasma levels afterlipid coprecipitate composition (Formulation AY-91-155,Example 1, Table 2) compared to a MCT oil solution of thedrug.FIG. 4 shows the oral pharmacokinetics in rats ofDexanabinol formulated in a solid TPGS lipidcoprecipitate(Formulation AY-91-155, Example 1, Table 2) atthree different dosages.â FIG. 5 shows the Dexanabinol rat plasma levels afteroral administration (30 mg/kg drug, n=5) of a powdered solidTPGS lipid coprecipitate composition (Formulation AY-175-3)prepared as described in Example 1 (Table 1).FIG.gastric fluid from commercial CoQ1O gelatin capsules and from6 shows the in vitro release of CoQ10 in simulateda powdered solid TPGS/PVP lipid coprecipitate formulation(ED-107-157, Example 5, Table 5) packed in gelatin capsule.-7-WO 98/08490101520253035CA 02264718 1999-03-01PCT/US97/15072DETAILED DESCRIPTION OF THE INVENTIONThis invention is directed to compositions comprising dry tocopherol polyethyleneglycol succinate solidcoprecipitates useful for the oral delivery of lipophilicsubstances, and to methods for preparing and using suchcompositions.Examples of lipophilic substances that exhibit poor oralbioavailability include lipophilic drugs, vitamins, andhormones. These include steroids, steroid antagonists,non-steroidal anti-inflammatory agents, antifungal agents,antibacterial agents, antiviral agents, anticancer agents,anti-hypertensives, anti-oxidants, antiâepileptic agents andanti-depressants among many others. Additional examples oflipophilic drugs with very poor water solubility and low oralbioavailability which could benefit from oral dosage formsare the neurohormone melatonin, the antifungal agentamphotericin B, the anticancer drug etoposide, as well astamoxifen and its analogs. More specific compounds includecannabinoids, as exemplified by dexanabinol, and vitamins,Preferredenzymes or coenzymes, as exemplified by CoQ10.lipophilic substances are those which have a water solubilityof <200ug/ml in water at room temperature (25°C), and morepreferably <50ug/ml. Depending on the inherent activity ofthe active ingredient its quantity has to be adjusted foreach specific drug. one of ordinary skill in the art candetermine the appropriate concentration by routine testing.Compositions according to the invention can contain, forexample, as the active lipophilic ingredient: steroids ornon-steroidal anti-inflammatory drugs, antibiotics,antifungals, antivirals, antihistamines, antineoplastics orimmunosuppressives. Specific examples would includesubstances such as clotrimazole, bifonazole, tetracycline,miconazole, triamcinolone, amphotericin, erythromycinhydrocortisone, iodoxuridine, diphenhydramine, minoxidil,lidocaine, tetracaine.For pharmaceutical use, the following categories ofdrugs are suitable: hypnotics, sedatives, anxiolytices,-8..WO 98/08490101520253035CA 02264718 1999-03-01PCT/US97/15072antidepressants, anticonvulsants, anti-inflammatory drugs,antiâfungals, prostanoids, prostanoid agonists, prostanoidantagonists, analgesics, hormones and vitamins. Specificexamples would include lipophilic peptides, barbiturates,benzodiazepines, phenothiazines, cyclosporin, diphenoxylate,tacrine, diclofenac, dexamethasone, prostaglandins,nifedipine, atropine, verapamil, fentanyl, lipophilicpeptides, ketoprofen, phenytoin, miconazole and ketoconazole.For vitamins, the active ingredient may be for exampleVitamin A, Vitamin E, a polyunsaturated fatty acid such aseicosapentanoic acid, retinoids, carotenes.For lipophilic hormones, a wide variety of activeingredients, including steroids such as estradiol,progestins, androgens and their antagonists are suitable forformulation according to the present invention.The compositions of the present invention are alsouseful for the enhanced oral availability of certain peptidesor proteins such as the hydrophobic or lipophilic peptidesexemplified by the immunosuppressive agent cyclosporin.The present invention discloses a novel way to increasethe oral bioavailability of any of these lipophilicsubstances. More particularly, the invention relates tosolid coprecipitates comprising the lipophilic substance, asurfactant vehicle having a melting point close to human bodytemperature, and at least one dispersion agent/adjuvant.After mixing with body fluids, such as gastric fluid,these compositions undergo quick dissolution with resultantmicelle formation or emulsification. A good example of asurfactant vehicle (which can quickly disperse drugcoprecipitates) is alphaâtocophero1 polyethylene glycolsuccinate (TPGS), which was first disclosed in US patent3,102,078.esterification of a hydrophilic polyethylene glycol moleculeTPGS is an amphipathic molecule, prepared by the(usually with a mean molecular weight 1000, and about 20-25ethylene oxide chains) with the carboxylic group ofTPGS isa water soluble compound (to 20% w/v) and forms micellarhydrophobic d-alphaâtocopherol hemisuccinate (acid).-9..101520253035CA 02264718 2005-01-11solutions with a critical micelle concentration (CMC) of0.4-0.6 mM/L (about 0.075%).balance (HLB) of TPGS is about 15-19.of TPGS, high HLB and water solubility, and low CMC make themolecule an excellent emulsifying agent for lipophilicThe hydrophilicâlipophilicThe amphipathic naturesubstances. The emulsification and subsequent increase insurface area of the lipophilic substance results in increasedgastrointestinal drug absorption and bioavailability.Toxicological studies have shown that TPGS is safe foringestion by humans as a dietary or nutritional supplement.In conclusion, TPGS can be safely used as a surfactant and abioenhancer for lipophilic substances of.limited absorptionin gastrointestinal region. Moreover, the antioxidativeproperties of TPGS improve the stability of TPGS containingformulations.According to the present invention, it now is disclosedthat solid TPGS coprecipitate formulations selfâemulsify whendispersed in a aqueous medium, like gastric fluid, formingvery small drug mixed-micelles. Lipophilic substances loadedin TPGS solid coprecipitate formulations will be absorbedmore easily by the gastrointestinal tract enhancing its oralbioavailability.The solid TPGS coprecipitates of the present inventionare composed of three essential ingredients: a lipophilicsubstance with low water solubility, tocopherolpolyethyleneglycol succinate, and at least one dispersionadjuvant.According to a more preferred embodiment of the presentinvention, the solid TPGS coprecipitates of the presentinvention, may advantageously further comprise a freeâf1owingimparting agent, such as fumed silica (Cab-OâSilâ, CabotUSA).Lipophilic substances incorporated in the solid TPGSCorp.,coprecipitates of the present invention have shownunexpectedly improved in vitro drug release in simulatedgastric fluid and enhanced oral bioavailability...10_101520253035CA 02264718 2005-01-11The lipophilic substance content in the solid TPGScoprecipitates of the present invention may range from about0.01-50% of the total solid weight of the composition, morepreferably in the range of about 5-30% of the total solidweight of the composition, and still more preferably about10-20% of the total solid weight of the composition.The content of TPGS in the finalformulations of the present inventionabout 5-65% of the total solid weightmore preferably in the range of aboutcoprecipitateis in the range ofof the composition,10-60% of the totalsolid weight of the composition, and still more preferably inthe range of about 10-50% of the total solid weight of thecomposition.The content of dispersion adjuvant in the finalcoprecipitate formulations of the present invention is in therange of about 5-75% of the total solid weight of thecomposition, more preferably in the range of about 10-40% ofthe total solid weight of the composition, and still morepreferably in the range of about 10-30% of the total solidweight of the composition. This dispersion adjuvant isincluded to facilitate the homogeneous dispersion of thelipophilic substance in the mixture.According to a preferred embodiment of the presentinvention, the solid TPGS coprecipitates of the presentinvention comprise a dispersion adjuvant selected from thegroup consisting of polyvinylpyrrolidone (PVP); a mediumchain triglyceride or MCT oil; a long chain triglyceride orLCT oil; tocopherol acetate; polyethyleneglycol; or otheradjuvant substance that can improve the dissolution of thelipophilic substance into the mixture or to help thelipophilic substance disperse homogeneously in the TPGS.Specific examples of such dispersion adjuvants include:(ISP Technologies, USA); Kollidonm 12 PF (BASF,Germany); Miglyolm 812 (Hulls, Germany); LCT oil (Croda);Carbowaxâ 1450 USA).The solid TPGS coprecipitates of the present inventionPovidoneâand(Union Carbide,may further comprise any suitable nontoxic carrier or diluentWO 98/08490101520253035CA 02264718 1999-03-01PCT/U S97/ 15072powder known in the art to serve as a free-flow impartingagent. Common examples of such additives are silicondioxide, starch, lactose, sucrose, sorbitol, talc, stearicWhen thecompound is formulated into a tablet or pill, the tablet oracid, magnesium stearate, and dicalcium phosphate.pill can be coated or otherwise compounded withpharmaceutically acceptable materials known in the art toprovide a dosage form affording prolonged action or sustainedrelease. The solid TPGS coprecipitates may also be preparedin gelatin capsules.According to a preferred embodiment, solid TPGScoprecipitates are further mixed with fumed silica, such asCAB-O-SIL® (Cabot Corp., IL., Us), which is a powderymaterial with extremely small particle size and enormoussurface area.Fumed silica can act as a dry lubricant, promoting thefree flow of the powdery mixture and preventing the mixturefrom caking or lumping. The free-flow, anti-caking andanti-clogging characteristics which are imparted to powders,granules and pellets by the addition of small amounts offumed silica are the result of several actions. For example,the submicroscopic size of the silica aggregates permits themto move easily between the larger particles of the other dryingredients, and in most cases fumed silica probably forms acoating on the powder particles. The fumed silica layer alsodecreases bulk tensile strength and shear strength, whileneutralizing the electrostatic charge on the particles.After blending with the other powders, fumed silicaadsorbs some or all the moisture which may be present in oron the product particles. The fumed silica aggregatesprevent other particles from contacting each other to formthe nuclei that would otherwise lead to the formation oflarger lumps and cakes. This spacing and lubricating actionhelps to keep materials moving through the apertures ofprocess equipment valves, spray heads, storage bin openings,bag and drum spouts, and aerosol nozzle orifices._12_WO 98/08490101520253035CA 02264718 1999-03-01PCT/US97/ 15072Most powdered materials can be kept free flowing byadding a concentration of fumed silica in the final productrange of about 0.5-25% (total solid weight).concentration can be determined by working up or down inThe optimumsmall steps. The more preferred weight percent of fumedsilica in the final product will be in the range of about1â20% (total solid weight).become caked can usually be rendered free flowing by blendingthem with fumed silica (about 2% of the total solid weight,Even powders which have alreadyor less).Products which cannot be processed beyond a sticky ortacky powder can be made free flowing by adding the properlevel of fumed silica as a final finishing step. Fumedsilica can also be used to promote free flow in spray-driedor freeze-dried products.In some cases it can be introduced into the originalemulsion, suspension or solution, or blended in later. Fumedsilica has also been used to coat powdered and pelletizedproducts to prevent them from caking later.The content of silicon dioxide in the final solid TPGScoprecipitates is commonly in the range of about 5-20% of thetotal solid weight of the solid coprecipitate, and morepreferably in the range of about 10-20% of the total solidweight of the solid coprecipitate.The preparation of solid TPGS powdered coprecipitates oflipophilic substances of the present invention may beprepared by different methods as described in the followingnon-limiting examples.-13.-101520253035CA 02264718 2005-01-11EXAMPLESExample 1: Preparation of tocopherol polyethyleneglycolsuccinate/polyvinylpyrrolidone powderedcoprecipitates of Dexanabinol byfreeze-drying.This example illustrates the preparation of tocopherolpolyethyleneglycol succinate/polyvinylpyrrolidone (TPGS/PVP)powdered coprecipitates of Dexanabinol by freezeâdrying. Thefinal weight/weight % composition of the differentformulations prepared were as described in Table 1.Table 1. Weight/weight % composition of TPGS/PVP/DexanabinolcoprecipitatesFormulationCode andIngredients AY-91-I59-6 AY-91-175-2 _ AY-91-175-2 AY-91-17$-I0 Av-91-175.3 AY-91-!75-4TPGS 54 so 40 so so 40Dexanabinol 6 10 20 20 30 30PVP 26 25 26 15 25 16Cab-O-sil" 14 14 14 :4 14 14TPGS (Eastman-Kodak Co., USA) was comelted with PVP(Kollidonm PF 12, added from a 10%water) at 40â60° C in a water bath.the melted materials and the mixtures were gently agitatedCab-O-Silâ (Cabot CORP., USA, addedfrom a 5% solution in water) was added and the mixtures wereBASF, Germany, solution inDexanabinol was added tofor several minutes.agitated again for several minutes. After cooling to roomtemperature, TPGS/PVP/dexanabinol coprecipitates were formed.The coprecipitates were freeze-dried overnight using a Christbeta lyophilize (Germany). Powdered freeâflowing Acoprecipitates quickly dispersible in water were obtained.TPGS/PVP/dexanabinol coprecipitates containingadditionally tocopherol acetate and Miglyolm 812 as describedin Table 2 were also prepared by freezeâdrying using the same-14-101520253035CA 02264718 2005-01-11method, which resulted in the formation of powderedfreeâflowing coprecipitates quickly dispersible in water.Table 2. Weight/weight % composition of TPGS/PVP/Dexanabinolcoprecipitates containing Miglyolm and tocopherolacetate.FunuhlicllCain andlnuufulu AY-9|-I6!-5 AY-91-175-5 AY-91-175-6 AY-91-175-7 AY-91-1'75-9 AY-9| -175-I AY~91-I65-9 AY-9|-155TPGS 13.4 13.4 13.4 13.4 13.4 13.4 7.5Touqiaswl 31.3 15.3 10.3 33.3 Z1.) 3.3 37.5Iillcmslyclâ 812 3.3 15.3 N3 I0.) 20.3 8.3 37.5 34Deumbincl 6 I6 16 16 16 30 - 9PVP 26 26 26 26 16 â- -Cab-0-5iJ" 14 H 14 I4 14 N â -The obtained powdered coprecipitates were filled in hardgelatin capsules (No. 1) for in vitro release experiments insimulated gastric fluids and for oral bioavailability studiesin rats (minicapsules, Torpac, NJ, USA).Preparation of TPGS/PVP powderedcoprecipitates of Dexanabinol by spray-drying.Example 2:This example illustrates the preparation of a TPGS/PVPpowdered coprecipitate of Dexanabinol by spray-drying. Thefinal weight/weight % composition of the formulation preparedis described in Table 3:Table 3. Weight/weight % composition ofTPGS/PVP/Dexanabinol coprecipitate formulationprepared by spray-dryingFormulation codeand Ingredients AY-91-175-3TPGS 30Dexanabinol 30PVP 26CabâOâSilm 14-15.-101520253035CA 02264718 2005-01-11TPGS (249 mg) was melted in a water bath at 50° C.Dexanabinol (249 mg) were then added to the melted TPGS andthe mixture was shaken for several minutes at 50° C. PVP(2.2 ml of a 10% solution in water) was added to the comeltedmixture of TPGS/Dexanabinol TPGS and the mixture was shakenagain for several minutes. Finally, CabâOâSilâ (2.3 ml of a5% solution in water) was added and the formulation wasshaken again for 1 hr at 50° C.The resultant formulation was spray-dried using a YamatoPulvis GA32 Mini spray-dryer. The drying conditions were:flow rate 7 ml/min, inlet temperature 130 C°, outletAhomogeneous and quickly water-dispersible dry powder of thetemperature 80° C, and drying air flow 0.45 n9/min.TPGS/PVP/Dexanabinol coprecipitate was obtained.In vitro release of Dexanabinol from solidlipid TPGS coprecipitates in simulated gastricfluid.Example 3:In vitro drug release of Dexanabinol from solid TPGSlipid coprecipitates was determined by placing a hard gelatincapsule No. 1 containing the formulation in 50 ml ofsimulated gastric fluid (150 mM Nacl, pH 1.2, 37° C,containing 1% Tweenâ 80 as sink). Gentle stirring wasprovided by a magnetic bar. Samples were drawn from therelease medium at prefixed time intervals and filteredthrough a 2.7 pm PTFE filter (Whatman).Dexanabinol was determined by HPLC using a Kontroninstrument equipped with pump, UV detector, and autosampler.A summary of the typical chromatographic conditions of themethod is provided below:Column: Merck 50980 supersphere 100RP-18, 75x4 mm, 4 pm.Mobile Phase: 30% phosphate buffer (0.01M Kaï¬wy, pH 3.0): 70%Flow rate: 1 ml/min.acetonitrile (v/v). Detectorwavelength: 280 nm. Injection volume: 20 pl. Columntemperature: ambient. Retention time: about 5.8 min. Runtime: about 9 min.-16-WO 98/08490101520253035CA 02264718 1999-03-01PCT/U S97/ 15072Figure 1 shows the in vitro release data for TPGS/PVP/Dexanabinol coprecipitate formulations in simulated gastricfluid.formulation, very good Dexanabinol release (from 60-95%) wasDepending on the specific composition of theobtained -- mainly during the initial 10-20 min.Example 4: Enhanced oral bioavailability of Dexanabinolin solid TPGS coprecipitate formulations.The solid TPGS powdered coprecipitates were filled inhard gelatin minicapsules (Torpac, NJ, USA) and tested fortheir oral bioavailability in rats. Male Sprague-Dawley rats(220-260 g, n=4) were administered orally with Dexanabinolformulated either as TPGS coprecipitates as described inExample 1 (Tables 1, 2, and 3) and filled in hard gelatinminicapsules (Torpac, NJ, USA) or in a MCT oil solution at10, 25, and 50 mg/kg doses. Blood samples were collected at0,0.5, 1.0, 2.0, 3.0, 5.0, 8.0,The samples were centrifuged at 10,000 rpm for 5 min and theand 24 hour time intervals.plasma was separated and stored frozen at -20° C until plasmaDetermination ofThechromatographic conditions were as described in Example 3.Dexanabinol levels were analyzed.Dexanabinol in the plasma was performed by HPLC.Plasma samples stored at â20° C were defrosted and diluted1:4 as follows: 150 ul plasma sample was transferred into a1.8 ml Eppendorf tube and 150 pl of acetonitrile were added.The sample was vortexed and centrifuged in a microfuge for 10minutes at 10,000 rpm. The upper clear liquid was thentransferred into an HPLC glass conical vial. A calibrationcurve was used for calculating Dexanabinol plasma levels.Corrections of sample peak areas were done by subtracting theof average value of the peak area obtained for samples atzero time (blank plasma).Figure 2 shows the oral pharmacokinetics of Dexanabinoladministered to rats (n=5) at a 50 mg/kg dose as a solid TPGSlipid coprecipitate (Formulation AYâ91â155 prepared asdescribed in Example 1, Table 2) compared to a solution of-17-101520253035CA 02264718 2005-01-11the drug in MCT oil (Miglyolm 812). A two fold increase inDexanabinol plasma levels was obtained with the coprecipitateformulation compared to the oil solution, demonstratingenhanced oral absorption of Dexanabinol from the solid TPGScoprecipitate formulation. This data indicates the goodwater dispersibility of the formulation, which, in turn,probably facilitates the uptake of the drug from thegastrointestinal tract.Figure 3 shows the rat oral pharmacokinetics ofDexanabinol from TPGS solid lipid coprecipitate formulation(Formulation AY-91-155 prepared as described in Example 1,Table 2) at three different doses.concentration (cmu) was obtained after 5 hours (tmu) for all10, 25,Figure 4 illustrates the oral pharmacokinetics ofA maximal drugthree doses tested, and 50 mg/kg.Dexanabinol administered to rats (n=5) at a 30 mg/kg dose asa powdered solid TPGS/PVP lipid coprecipitate in hard gelatinminicapsules (Formulation AYâ175-3 prepared as described inExample 1, Table 1).levels of the drug and demonstrating the improved deliveryThe figure shows increased plasmaand oral bioavailability of Dexanabinol.Preparation of solid TPGS lipid coprecipitatesof Ubiquinone (coenzyme Q10).Example 5:This example illustrates the preparation of tocopherolpolyethyleneglycol succinate coprecipitates of Coenzyme Q10.The final weight/weight % composition of the differentformulations prepared is described in Table 4:- 13 -CA 02264718 2005-01-11Table 4. Weight/weight % composition of TPGS/CoQ10coprecipitates.Formulation Codeand Ingredients 106-193 106-19C 106-16A 106~llC5 was 20 21 47.1 47.5Coenzyme Q10 5 5 5.8 5Mcrml 75 â ~ âLCT oil -â 74 -- -PVP â â 411 â1 0 PEG 1450 - _ - 47.5Ubiquinone (Coenzyme Q10) was obtained from GlobalMarketing Associates, Inc. (San Francisco, CA). Thepolyethyleneglycol used was Carbowaxâ PEG 1450 from Union15 Carbide (CT, USA). The MCT oil (medium chain triglycerides)used was Miglyolâ 812 (Hulls, Germany). LCT oil (long chaintriglycerides) was from Croda. TPGS/CoQ10 coprecipitateswere prepared by heating together all the components at50°â60° C with subsequent cooling to room temperature.20 Compositions containing polyvinylpyrrolidone (PVP K-12) wereprepared using 50% PVP solutions in absolute ethanol anddrying the obtained mixture at 40°-45° C for 4-6 hours. Theobtained TPGS/CoQ10 coprecipitates showed quick waterdispersibility with rapid release of the CoQ1o.25Example 6: Preparation of TPGS/PVP powderedcoprecipitates of Ubiquinone (coenzyme Q10) byfreeze-drying.This example illustrates the preparation of tocopherol3° polyethyleneglycol succinate powdered coprecipitates ofCoenzyme Q10 by freeze-drying. The final weight/weight %composition of the different formulations prepared isdescribed in Table 5:35- 19 -101520253035CA 02264718 2005-01-11Table 5. Weight/weight % composition ofTPGS/PVP/CoQ10 powdered coprecipitates.Formulation Codeand Ingredients so-107-157 an-107-155-1 ED-107-I55-B AY-91-I75-Cwas 40 56 so asCoenzyme Q10 9 I3 1 I 15mm 41 ï¬ u _Cab-O-sil" no 15 14 19TPGS (Eastman-Kodak Co.,a water bath.1-2 hr.again for several minutes.TPGS/PVP/CoQ10 coprecipitates were formed.CabâO-SilâUSA) was melted at 40°-60° C inCoenzyme Q10 was added to the melted TPGS andthe mixtures were gently agitated for several minutes. PVP(Kollidonm PF 12, BASF, Germany) was then added from a 30%solution in water and the mixtures were shaken at 40° C foradded from a 5%solution in water) was added and the mixtures were agitated(Cabot CORP. USA,After cooling to room temperatureThecoprecipitates were freeze-dried overnight using a Christbeta lyophilizer (Germany).Powdered free-flowingcoprecipitates quickly dispersible in water were obtained.They were packed in hard gelatin capsules for in vitrorelease studies. =In vitro Release of Coenzyme Q10 from powderedTPGS/PVP coprecipitates in simulated gastricfluid.In vitro drug release of CoQ10 from a powdered TPGS/PVPcoprecipitate formulation and from a commercial productcontaining equivalent amounts of CoQ10 were determined byplacing a hard gelatin capsule in 50 ml of simulated gastricfluid (150 mM Nacl, pH 1.2,sink.37° C) containing 1% Tweenâ 80 asGentle stirring was provided by a magnetic bar.Samples were drawn from the release medium at prefixed timeintervals, filtered through a 2.7 pm Whatman GF filter andanalyzed for CoQ10 concentration.-20..101520253035CA 02264718 2005-01-11Figure 5 shows the in vitro release patterns of CoQ10from a powdered TPGS/PVP coprecipitate formulation comparedto a commercial product ENERGYCO® CoQ10 (Herbamed-AssutechLtd., Rehovot, Israel) in simulated gastric fluid.CoQ10 was determined in the commercial product, thepowdered TPGS/PVP coprecipitate formulation, and in therelease medium in vitro by extraction with Dole reagent(isopropanol:heptane:water, 45:36:17) by measuring theCoQ10samples (0.5 ml) were added to 3.5 ml of Dole reagent andabsorbance at 270 nm using a calibration curve.mixed thoroughly. The two phases were then allowed toseparate for 10 min at room temperature. CoQ10 was extractedselectively in Dole heptane upper phase, which wastransferred to a quartz cuvette for absorbance measurement.The % release of CoQ10 from the marketed product wasvery low compared to a very quick and significant releasefrom the powdered TPGS/PVP coprecipitate formulation. EachENERGYCO® CoQ10 hard gelatin capsule contains 50 mg of CoQ10mixed with rice powder. After capsule disruption in thesimulated gastric fluid, big aggregates or clusters of CoQ10and swelled rice powder were observed. These observationsmay explain the low CoQ10 dissolution into the releasemedium. Since particle size is a determinant factor in therate and extent of drug absorption from the gastrointestinaltract, this result indicates low oral bioavailability ofCoQ10 from the commercial product compared to powderedTPGS/PVP coprecipitate formulation of the present invention,which is quickly dispersible in the simulated gastric fluid.Preparation of TPGS/PVP powdered coprecipitateof Melatonin.Example 8:TPGS (500 mg) was melted at 40°â60° C in a water bath.Melatonin (100 mg, from Jansenn Chimica, Belgium) was addedto the melted material and the mixture was agitated forseveral minutes. A small amount of absolute ethanol (up to5%, v/v) was added in order to get an homogeneous mixture.PVP (Kollidonâ PF 12, 2.6 ml of a 10% solution in water) was-21-.101520253035CA 02264718 2005-01-11then added and the mixture was agitated again for severalCabâOâSilâ (2.8 ml of a 5% solution in water) wasadded and the mixture was agitated again for several minutes.The resultant TPGS/PVP/melatonin mixture was thenminutes.freeze-dried overnight using a Christ beta lyophilizer(Germany). A powdered free-flowing TPGS/PVP/melatonincoprecipitate quickly dispersible in water was obtained.Preparation of TPGS/PVP powdered coprecipitateof Amphotericin B.Example 9:TPGS (500 mg) was melted at 40°-60° C in a water bath.Amphotericin B (100 mg, from Dumex, Denmark) was added to themelted material and the mixture was agitated for severalminutes. A small amount of absolute ethanol (up to 5%, v/v)PVP2.6 ml of a 10% solution in water) was thenwas added in order to get an homogeneous mixture.(Kollidonm PF 12,added and the mixture was agitated again for several minutes.CabâOâSilâ (2.8 ml of a 5% solution in water) was added andthe mixture was agitated again for several minutes. Theresultant TPGS/PVP/Amphotericin B mixture was then Vfreezeâdried overnight using a Christ beta lyophilizerA powdered free-flowing TPGS/PVP/Amphotericin Bcoprecipitate quickly dispersible in water was obtained.(Germany).Preparation of TPGS/PVP powdered coprecipitateof Tamoxifen methyliodide.Example 10:TPGS (500 mg) was melted at 40°-60° C in a water bath.Tamoxifen methiodide (100 mg, from Pharmos Corp., FL, USA)was added to the melted material and the mixture was agitatedfor several minutes. A small amount of absolute ethanol (upto 5%, v/v) was added in order to get an homogeneous mixture.PVP (Kollidonm PF 12,then added and the mixture was agitated again for severalCabâOâSilâadded and the mixture was agitated again for several minutes.The resultant TPGS/PVP/Tamoxifen methiodide mixture was thenfreeze-dried overnight using a Christ beta lyophilizer2.6 ml of a 10% solution in water) wasminutes. (2.8 ml of a 5% solution in water) was-22-101520253035CA 02264718 2005-01-11(Germany). A powdered freeâflowing TPGS/PVP/Tamoxifenmethyliodide coprecipitate quickly dispersible in water wasobtained.Preparation of TPGS/PVP powdered coprecipitateof Etoposide.Example 11:TPGS (500 mg) was melted at 40°-60° C in a water bath.Etoposide (100 mg, from Sigma, St. Louis, USA) was added tothe melted material and the mixture was agitated for severalA small amount of absolute ethanol (up to 5%, v/v)PVP2.6 ml of a 10% solution in water) was thenminutes.was added in order to get an homogeneous mixture.(Kollidonm PF 12,added and the mixture was agitated again for several minutes.Cab-O-Silm (2.8 ml of a 5% solution in water) was added andthe mixture was agitated again for several minutes. Theresultant TPGS/PVP/Etoposide mixture was then freeze-driedovernight using a Christ beta lyophilizer (Germany). Apowdered free-flowing TPGS/PVP/Etoposide coprecipitatequickly dispersible in water was obtained.Preparation of TPGS/PVP powdered coprecipitateof Clycosporin A by freeze-drying.Example 12:This example illustrates the preparation of tocopherylpolyethylenegylcol succinate powered coprecipitate of thepeptide drug cyclosporin A, a very lipophilic undecapeptidewith very low water solubility and poor oral bioavailability,by freeze-drying. The final weight/weight % composition ofthe formulations prepared was as described in Table 6:-23-CA 02264718 2005-01-11Table 6. weight/weight % composition ofTPGS/PVP/Cyclosporin A coprecipitateformulation prepared by freeze-drying.Formulation code5 and Ingredients ED-119-148TPGS so ICyclosporin A 10 "PVP 26 ICabâOâSilâ 1410All lipid components and cyclosporin were dissolved inethanol. CabâOâSilâ was added from a 5% water dispersion tothe ethanolic solution (at a final ethanolzwater v/v ratio of1:1) and the mixture was shaken at 40C for several minutes.15 The mixture was then frozen at â30%:for several hours andlyophilized overnight using a Christ lyophilizer (Germany).Homogeneous dry quick-dispersible power containing thecyclosporin-lipid mixture was obtained.It will be appreciated by the artisan that many20 additional modifications or variations of these compositionsare feasible. The scope of the invention is, therefore, notto be construed as limited to the foregoing examples, but isto be construed, at the very least, to be commensurate withthe scope of the following claims.25 -3035-24-
