AU1131799A - 'Composition and use for reducing adenosine levels and treating asthma and other diseases and conditions afflicting the airways with an epiandrosterone, a ubiquinone, or both.' - Google Patents

Description

AuSTRAL2IA patents Acz 194 COMPLETE

SPECIFICATION

STANDAFRD

PATENT

Applicant: EAST CAROLINA

UNIVERSITY

invention Title: GO RAIGLWLVL FAEOIEO METHOD OF PREVENTINGO TETNGLWLVSOF DNSI

R

ADENOS INE DEPLETION The followinlg statemenlt is a full description of this inventijon, includinlg the best method of performinlg it known to me/us: METPOD 0;7 DREVENTTNG OR TRrATTNG LOIN LEVELS OF ADENOSThN.E OR ADE NOS7NE DEPLETION The ent~ire disclosure in the complete snec2.tlcation of our Australian Patent zkoplicattIo No.

41867-7/96 is by this cross-reference incoroorated into the oresent seeclitcatLol.

Field of the invention This inveniIon concerns methods of" treating adenosine deoletion by the administration of folinic acid or a oharmacauticallv acceotable salt thereof. This invention furth-er concerns methods of' treating asthmia by administering dehydroepiandrosterone, aralogs thereof, or 15 the oharmaceutically acceotable salts thereof.

Background of the Invention Adenosine is a Durine which contributes to intermediary metabolism and participates in the regulation 2 20 of ohvsiological activity in a varietv of ma-mnalian.

tissues. A4denosine oarticipates in manx' local regulatory mecnanisms, in particular svnaoses in the central nervous system (CNS) and at neuroeffector junctions in tne peripoheral nervous system. in the CNS, adenosine: lnlhi-s the release of a variety of neurot-ransmitters, such as acetvlcholine, noradrenaline, dopamine, serotonlen, glutamate, and GABA; depresses neurotransmission; reduces neuronal trrlng to induce spinal analgesia; and possesses anxiolyvtic properties. See A. Pelleg and R. Porter, Pharrnacotherapy 10(2). 157 (1990); J. Daval, et al., Life Sciences 49:1435 (1991). in the heart, adenosine suppresses pacemaker activity, slows JAl conduction, possesses antiarrhvtrnpic and arrhythmogenic effects, modulates autonomic control, and triggers the synthesis and release of prostaglandins. See K. Mullane and M. William, Adenosine and -Adenosine Receptors p. 289 Williams, ed. Humana Press, 1990). Adenosine has potent vasodilatory effects and 46

OR

-3rac.UlatCes ,,as;cujar tone. S~ee A DeVsea ez a. J. pfge

Z-:

Ah.406: 608 1986) Adenlosine is CII fl ben ue clinically for the treatment Of s~aet,,'Ia !tachycardia and ether cardiac anomalies. See C. Chroni)ster, _InZerlca JournaI of'~t caCare 41-4:7(93) .A de-noslfe analogues are boeing invetigated o- use as a-nticollXuisant, anxiolv-tic anod neuroprotective agents. see, M. iig~fS e al pamCV Wor0-d Scien~ce 16 62-68 \denosi~ne has also been imol1-icated as a primary determinant underlying the smPtOmfs oE bronchial asthnma. It induces bronchoconstriciof and the contraction of airl,,:y smooth muscle. See J. Thorne ana roadlev, A-mer~cal J ourna a~ s itorY &crtiCa 1 care MNedir-;Ie !1A9(2 pt.

392-399 (1994); S. Ali et lAel CifS3 (304) -165-167 (1992) Adenosine causes bronchoconstriction in asha-c u not in non-asthatics. See Bj orck et al., Anr 1astiat eis but Repr-1rY is-ase 145 !087-109l (1992);1 S. Hiolgarte et al-, Ar-M.ls Of:teN~ or ~dm Of Sciences '029: 227-236 (1991).

in viie' of the foregoing, i t wqill be readily a -iret that: e dnoil de-plet1ion can lead to a broad variety of deleterious cond Itios n ht toso treating adenosi-ne depletjon can be an ext-remelY- useful means of therapeutic intervention; and (i4) methods of inducing adenosine depletionl can also be -zsetul in treatinlg conditions such as asthma.

Throughout the description and claims Of this specification, the word -comprise" and vari-ations of the word, sich as c.omprising" and "cmvprises', means including but not limited to" and is not intended to exclude other additives, componlents, intege-rs or steps.

Summary of the invention Accordingly, a first aspect of the present invention provides an in vivo method of preventing or treating low levels of adenosinel or adenosifle depletion,

I:

3 .1 comprising administering to a subject in need of such treazment a composition comprising an adenosine increasing ffective amount of an agent selected from the group consisting of folinic acid hysiologically acceptable salts thereof and mixtures thereof.

The method comprises administering to the subject folinic acid or a pharmaceutically acceptable salt thereof in an amount effective to treat the adenosine depletion.

The method may be carried out on subjects afflicted with steroid-induced adenosine depletion, subjects afflicted with anxiety, subjects afflicted with a wasting disorder, or subjects afflicted wich any other disorder attributable to adenosine depletion, or where an increase in adenosine levels would be therapeutically beneficial.

A second aspect of the present invention provides a composition, comorising an agent selected from the group consisting of folinic acid, physiologically acceptable salts thereof and mixcures thereof.

A third asoect of the present invention is a method of treating asthma in a subject in need of such treatment by administering to the subject dehydroepiandrosterone, an analog thereof, or a pharmaceutically acceptable salt thereof, in an amount effective to treat asthma.

A fourth aspect of the present invention is the use of dehydroepiandrosterone, an analog thereof, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating asthma in a subject in need of such treatment.

Folinic acid is an intermediate product of the metabolism of folic acid; the active form into which that acid is converted in the body, ascorbic acid being a necessary factor in the conversion process. Folinic acid has been used therapeutically as an antidote to folic acid antagonists such as methotrexate which block the conversion of folic acid into folinic acid. Additionally, folinic acid has been used as an anti-anemic (combatting folate \Ise~a~il^~S\3cOE\seps~~c\Salf-j :1:liT deficiency). See The Merck Index, Monograph No. 41.41 (11th Ed. 1989). The use of folinic acid in patiencs afflicted with adenosine depletion, or in a method to therapeutically elevate adenosine levels in the brain or other organ has heretofor neither been suggested nor described.

Detailed Description of the Invention The method of treating adenosine depletion disclosed herein can be used to treat steroid-induced adenosine depletion, to stimulate adenosine synthesis and thereby treat or control anxiety in treating premenstrual syndrome); to increase weight gain or treat S' wasting disorders; and to treat other adenosine-related pathologies by administering folinic acid. Thus the term "adenosine depletion" is intended to encompass both conditions where adenosine levels are depleted in the subject as compared to previous adenosine levels in that subject, and conditions where adenosine levels are essentially the same as previous adenosine levels in that subject but, because of some other condition or alteration in that patient, a therapeutic benefit would be achieved in the patient by increased adenosine levels as compared to previous levels. Preferably, the method is carried out on patients where adenosine levels are depleted as compared to previous adenosine levels in that subject. The present invention is concerned primarily with the treatment of human subjects but may also be employed for the treatment of other mammalian subjects, such as dogs and cats, for veterinary purposes.

Folinic acid and the pharmaceutically acceptable salts thereof (hereafter sometimes referred to as "active compounds") are known, and can be made in accordance with known procedures. See generally THE Merck Index, Monograph No. 4141 (11th Ed. 1989); U.S. Patent No. 2,741,608.

Pharmaceutically acceptable salts should be both pharmacologically and pharmaceutically acceptable. Such pharmacologically and pharmaceutically acceptable salts can i\nc~5a~\~foeS\?irr~i\gsrp\jprci\la6~7-) l)iOi133 -f beormaedas aa Cmalor alk-aline eacn salts, such as sodium, pot-assaurn, or calcium salts, of the carboxylic C -L grout) of 'Olin -C acid. The calciu m salIt of fo~ acid is a Dp eferred pharMaceut-caIly acceptable salt.

The active cozmpounds are preferabliv- admiflistered to the subject as a pharmaceutical cotlmposition PnamaCutcalcomOSt4~l on or use in Lhe present 'nveaiton include those suitable for. inhalation, Oral, too-ca 1 (cludifl9 buccal, sublingadra n intraocular) parente-ra (includiing subcutaneous, intradermnal, intramuscular, intravenous and intraarticular) anci transiermfal adminstrationl. The compositions may conveniently be presented in ur i r oa- fomn may be prepared by any of the methods w'.

1 lknowan in the art.

suitable fo oral adinsrto mray be presented in discrete un-its, such as capsules, cachets, lozenges, or tablets, each cozitaining a predetermined a-mount of th e active compound as a Powder or granules; as a solution or a suspension in an aqueous or Pon-~aq~ueous icquid; or as an o 4 indater or water-in-Oil emuls~in Such compositions& may be prepared by any suitable method of pharmacy whichb includes the step of bringing into association the active compound and a suitable carrier. In general, the compositions of the invention are prepared by uniformlY and intimately admixing the active compound with a liquid or finely divided solid carrier, or both, anda then, if niecessary, shaping the resulting mixture. Eor exape a tablet may be preLpared by compressing or molding a powder or granules containi~ng the active compound, optionally with one or more accessory ingredients.

Comnpressed tablets may be prepoared by compressing, in a suitable machine, the compound in a free-flowing Form, sucn as a powder or granules otionally mixed with a binder, lubricant, inert diluent, anid/or surface. active disoersing agent(s). Molded tablets may be made by molding, in a suitable machine, the powdered compound moistened with an inert liquid binder- Compositions for oral administration oo na1 nclud- e en~teric coatings nn"n~~ art o ore-ventz degradation of h composizions in the sztomtach and orovide release ot the d-rug in the small intestine.

Compositions sultable for buccal (sub-lingual) ad7miStrationi include lozenges comprising the active comnourci -4 a flav.ored base, usually sucrose and acacia or tragacanth; and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacia.

suitable for oarenteral acartinstration comprise sterile aaueous anc non-aqrueous i ,-:ect-ion solution's of the active compound, which preparacions are preferablyv isotonic with the blood of the intended recipient. These preoarations may contain antioxidants, buffers, bact~eriostats and solutes which render the compositions isotonic with the blood of the intended recir~ent- _Aaueous and -non-aqrueous sterile suspensions may include suspending agents and thickening agents. The co-mpositions may be presented in unit-dose or multi-dose concaners, ro-r example sealed amooules and vials, and may be stored in a -Freeze-did (lyophilized) condition requiring only the addition of the sterile liouid carrier, 4Zor example. saline or water-forinjection immediately prior to use, Extemporaneous injection solutions and suspensions may be nreoared from sterile powders, granules and tablets of the kind previously described- Compositions suitable for topical application to th e ski--n preferably take the form of an ointment, cream, lotion, poaste, gel, spray, aerosol, or oil. Carriers which may be used include Vaseline, lanol-ie, polyethylene glycols, alcohols, transdermal enhancers. and combinations of two or more thereof.

Compositronrs suitable for transderma 1 administration may be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. Compositions suitable for transderimal administration may also be ne~tvrea v ioono-esis (see, hkCUZC.

esa-h3, s1 i s6) and ca!l take Cthe otma an oorio-nally buffered aqueous solution of t-he act~-ve cornround.

Dosage wsill %var'; dependirn on age, weight, and conni-'ion of: tne subject. Treazmenrt r-av be in- -i.a"td %:.ich small- dosages less than optim,,um dos e and increased until tne cotmum erect under the c~rcum.stances is reacnet.

general, the dosage will b e f: r 5 10 o r 20 mg /kg subecz: body weight, u-p to 100, 200, 500 or 1000 mgc,/kg subi-ect: body w-%eight. Currently, dosages of from 5 to 500 mg/'%g are preferred, dosages of from 10 to 200 mg/kg are more -Orezerrea, ana dosages of from 20 t o 100 mg/kg are mIost preferred. in general, the active campounas are oreferablv ad-milnistered at a concentration thac -will afford ezzective zesult s without causing ary undulv harmful or eeleterous side ef'fects, and can be administered eitn-er as a single unit dose, or- if desired, In convenient subunits administered at suitable times throughout tne day.

Also disclosed herein isa method o-f etn asthma, uarticula-rly n-on-steroid dependent ast'n-ma, bv administering to a subject in ned of such treatm-Lent dehvd CLO-'aroeoidro- terone (DiiEA) ,an analog therecf, or a onar.maceuticaliv accepotable salt thereof, in an amount ~fctie t inibit control asthma to that subject.

Examolas of D-1HEA -and analogs thereof that may be used to carryV out this method are represented by the formula: Rio wherein.* h-rcen, s sc_ mf is a Ohs'- 0 The en a-o atoo q:r. 04: aor 7a~ T z oreserit inth-e aloh-a or beta configu-rat~oOf r rie oz.fb cori ses a m~xture of can f igraA-OiS Ccounczs i11ustrat-ive of Fo=t,"Ila UI) abo-ve iclude:

DB,~~

ad is-- hre and ne double bond s presenz: 1 6 wlh-fUOOClaieoser.e isF P.1 is H arid the double bon~d is oresent; eticch-oianolofle, -wl-ere~in _R and IRI are each.

hydrogen anda Lthe doule bond is absent; SO:OM alnd is a suiphaiC groCup as d e =above, and, te double boned iS a bs e n t ?refeably, in "'ne co-mpoun-d of forMUla T, R s a!logen e. .b Ilo'. chlOro, _z C2o rO), ~s 7-vdr og ea, andn-the doutble bond is prsnt os referal h Sojp o-r 7or:tn, ,a TL are mane in accorna-le w:nkn O rocedur-es or variations thereof th-at be io apaen o those skilled in the art. Seeu.S. PatenIt No.

4,95,~55 U~ atent No. 2.240,472, EPO Patent ?Appln No- 429,37,POTPatnt ooln No. 91/0A03; see also M.Abou- Gharb; o er j. Porm ci 70, 115A-1157 (1981) iderck TndeX, grapn No. 771C0 (11t7h ed. 19S89) The compounds used to treat asthma nay be a&dLinistered per se ori te- -For of pharmfaceuticall_ *acceotable salts, as discussed above (th--e t-wo togetfl.ef again being -Rererred to as "activ-,,e compounds") The activie Comp~ounds Salts -may e adminhistered eithner svstemicalvY as 20 discussen above, or t o the lungs or subject1 as discussed below. In general, thae active compounds salts are administered in a dosage of 1 t1o 3600 mg/k,%g body egt more o~efer-ably about 5 ro 1800 mg/kg, and most preferablYv about 20 to 100 mg/kr-g. Th-e active compounds may be administered once or several times a nay.

phe active comtpounds discosd herein m~ay be acL-mn-istered to the lungs of a subject by any suitable means, but are preferablY administered by generatilng an aerosol comorisen of respirable pam-ticles, the rasipirable Particles comprised of the active comound, which particles the subj ect, inhales (i by i'-nhalation admini2stration).

TPhe resorable particles may be liquid o-r solid..

Particles comrprised of active compound for practicing the present invention should included particles of resoirable size: that is, particles of- a size,sufficiently small to pass through the mouth and larynx upon inhalation and into the bronchi and alveoli of the In ge-~ral~ .ce rang -ro-r' abc'u 5 1C ~ucS; z:e) are of O ZU es -abI -4e ;*sC.are ~eQ -he aerc',sQ. z:o.

d e z o S i z-he =--zcaz e a ~-a~icec 2 0~ CS2- e c0:n :ne range a q:ud chrnc<3.a ofc~ODSa acivC co:" nea:v con an :C~evn.l~ such ooo:rs rc~2-j eeD~D~ rr Iara.z ec mcro e actv copu may 'be prepared by r, a27, Z:ne :a2croflIZ2O ccrnmos2..f =n~gLa U2Czhsre2 bDreazk ucp or secaurate cu- &g ac±ma~5 S02ol oart2.culate comooslz:on COM pr'-SeC oz :he a:7Ccompoud 230 m,,ay o- ionailV contalca -adiera~ :hcsre Lac---cate he 07mccnc an aeoo di soersan-t :s laccoCe m ay cbe CC~- zn ac::vCe c 0o ul i.d-n any, su-,i"able ra:1O (eKc:, a 7-0 razo by weigh-t) AeosolS of ra- dq~ pa-r2.c',--s c rp r i 3-4 g Ile actwie Compound may be produced by any suitable means, sucn as -with a flCDUji-e ze, U.S. Pazenrt No. 4,501,729.

WNebuier arecocr~~~ avj:able de-vi ces -shl ch trn~rasojLutlofs or- sSp'ef-slOfls of ch-e ac:-11 Ing en: into a thte-apeu--c aerosoL misz eih- 'o means of acceler-azio, of: a coznn-ressed gas, o-ca- oxcgen.

.througn a na--ro,,w; veznturi r' ce or- by means of~ ulraonic aoickatiofl, Suitable comnos-4ti-ons use nebu-,j'z.ers con~s-i- af the ac~ein-g--d-elt 4- a 1Ucum are, d active ingredl-i-az co-Mp' ilg U-0 to 40% w/l.1 of zhe cmposit-ns, but pxreferab-y less 20% wiw/. Thecarrier is tvoicaliy wate-r a dillute acrueous alco-hol-csolution, preferable made isotonic with body fluids by the addition of, for example, sodium chlorid Optional additives include preservat~ves iE the comoositions is no", Prepared sterile, for example, methyl hydroxybenzoate, antioxidOants, flavoring agents, volatile oils, buffering agents and surfactants.

Aerosols of solid narticles comoris ing the active compnound mPayv likewise be oroduced with any Soli4d tparticulatze medicament aeroso; generator. Aerosol gezterat ors for administering solid partcicluate medicaments to a subject produce particles w%!hch are respirable, as explains above, and generate a voueof aerosol con~taining a oredetermined metered dose of medicamenc at a rate~r suitable for numan adM~i~StrC:jo_;:. -Examples of such aerosol ID~ generators include metered dose inhalers and insufflators.

Ubiquinone may be administered concurrently/ with the DHEA or analog thereof in the methods of treating asthma described above. lhe nobrase "concurrently administering," as used herein means that the DHEA or the 20 DHiEA analog are administered eit*-er simultaneously in time (preferably by formularting the two together in a common pharmaceuatical carrier), or at: different times durini h course of a common treatment schedule. In the latter case, the two comuounds are administered at times suf-ficiently close for the ubiqui-none to ubicuinone depletion in the lungs (and heart) of the subject and thereby counterbalance anyv deterioration of lung (and heart) function that may result from the administration of the DHEA or the analog thereof. The term "lubiauinone", as used herein, r-efers to a fairdly of- compounds having structures based on a 2 nucleus wvith a variable ternenoid acid chain containing on to twelve mono-unsaturated trans-isoporenoid units. Such compounas are knownm in the art as "Coenzyme Qn", in which n equals 1 to 12. These compounds may be referred to herein as compounds represented by the formula: iv 13J1,33 -13 C H 0 1 (CHiCH=CCH-)1..-H 0 rn to 10. -rfrby n the method o the nvetio, te u~ounone Is a compound according to formula given above, .,.herein n =6 to 10 coenz-vmes Q...and most p-referably wherein 10 Coenzymie Wh1ere the lubiquinone is formulated xith a pharmnaceutically acceotable carrier separatel y from the DHE'%. analog thereof, or salt thereof wvhere the DHEA analog thereof or salt thereof is aCL-inlstered to the lungs of the subject, and the ubic :uinone is administered systemicallv) it may be formulated bv any of the techniques set fforth above.

In general, the ubiauinona is adiministered in an amount effective to offset ubiauinone deoletion deuletion in the lungs and hearz of the subject induced by the DHEA, analou thereof, or salt thereof, and the dosage will vary dep~endina upon the cozd.ltion of the subject and the route of administration. The ubiauinoneis prefera1i, administered in a total amount per day of about 1 to 1200 mg/kg body %,eight, more preferably about 30 to 600 mg/kg, and most preferably about 50b to 15 gk.Teuiunn may be administered once or several tim-Tes a day.

T.he following examples are provided to more fully illustrate the roresent invention and should not be construed as restrictLive thereof. in the following examples, DHEA means dehydroepiandrosterone, s mneans seconds, mg means mil~ligrams, kg means 'Kilograms, kw means kilowatts, IN{z means megahertz, and fnaol means nanomoles.

EXAMPLE 1&2 -14 Ef fects oE Folinic Acid and DHEA on Adenosine Levels in VIVO Young adult male Fischer 344 rat-s (120 graris) were administered deh\',droEp-,ian-drcosr-erone (3H-P) (300 mgl/kg) Sor methv1:-eszosterone (40I- mg/kg) in carboxvuiethvlcellulose by gavage once daily for fourteen days, FoL3inic acid mg/k"g) w:as acbni-nisceredif, aeftnal once daily for fourLeen cz avs. On the Eirteent*- day, the animals wr sacrificed bv micro!w.ave pulse (1.33 kT1, 2:50 M'-7 6.5 S) to he craniu-,. which isn1vdenar.-res all brain zorotein **and prevents further metabo 1 i-s~n or adenes-ne *4earrs w-ere removed from ani-lals adflash rvli liquid !4zroje:! I wlih;n 10 seconds of- .ea.h Iiv swrrmvde bl~oc ar'd -Iash froze' i, i 32 seconds of dezz-h. Brain tissue a subseauenzlv dIissecr-ed. -issue ~osn was exLraczted. derivazized z.o e. NI 11hnadnsn Cn a naIv'Ye d by high performance .qicroanahv(?L)Using speccrof-luoromezric neco according zo cae rethod of Clark and Dar of Y ce'" (1988)) Resuts of iese exper imen s rsuaiedfTblI below... Resuk'Is are expressed as 7,e mean SE% with p<0-05 compared zo contirol group and p<10.105 comnpared Lo DHEA or mehvltesetoszercne zreatedz groups.

TABLE 1. Effects of DHEA. 6-1-methyltestosterone and fclinic acid on adenosine levels in various tissues of the rat.

Treatment jIntracellular adenosine (nmols)/mg protein H[eart Liver JLung Brain Control 10.6 ±0.6 14.5 ±1.0 13.1 ±0.2 0.5 ±0.04 n=1 2) I(n=1 2) J(n=6) (n=1 2) DHEA j6.7 ±0.5 16.4 ±1.4 2.3 ±0.3 0.19 ±0.01 (300 mg/kg) (n= 1 2 1y (n=12) (n=6)Wi (n=1 2)xV Methytestosterone 8.3 ±1.0 16.5 ±0.9 N.D. 0.42 ±0.06 mg/kg) j(n y 15 I Methyltestosterone (120 mg/kg) Folinic Acid mg/kg) 6.0 ±0.4 (n=6)yp 12.4 ±2.1 (n=5) 5.1 0.5 (n=6)xi 16.4 ±2.4 (n=5)

N.D.

N. D.

I I DHEA (300 mg/kg) 11.1 ±0.6 Folinic Acid (n=S)p mg/kg) Methyltestosterone 9.1 ±0.4 (120 mg/kg) (n=6)0 Folinic Acid mg/kg 18.8±t1.5 (n=5)o

N.D.

0.32 ±0.03 (n=6>i' 0.72 ±0.09 0.55 ±0.09 0.60 ±0.06 t N. D. IN.D.

Trhe results OEf these experimentE; indicate that rats administered DH-EA or methyltestosterofle daily "-or two weeks show,.ed multi-organ depletion of adenosine. Depletion was dramatic in brair (60% deoletion for DHEA, 34%10 for high dose methyltestosterone) and heart (37% deoletion for DHEA.

22% denlet-ion ror high dose methvltestoscerone) Coadministration of folinic acid completely abrogated steroid-mediated adenosine dezpletion. 7Folinic acid administered alone induce increases in adenosine levels for all organs stdied.

The foregoing examples are illustrative of the present invention, and are not to be construed as limiting thereoF. The inv.ention is defined by the following claims, with equivalents of the claims to be included therein.

Claims (47)

1. An in vivo method of preventing or treating low levels of adenosine or adenosine depletion, comprising administering to a subject in need of such treatment a composition comprising an adenosine increasing effective amount of an agent selected from the group consisting of folinic acid physiologically acceptable salts thereof and mixtures thereof.

2. A method according to claim 1, wherein the low 1 0 levels of adenosine or adenosine depletion are associated Stwith a disease or condicion selected from the group S. consisting of steroid administration, premenstrual syndrome, wasting, anxiety and other adenosine-associated pathologies.

3. A method according to claim 1 or claim 2, wherein the low levels of adenosine or adenosine depletion are associated with he administration of steroids to the subject.

4. A method according to claim 1 or claim 2, wherein 20 the disease o condition is associated with pre-menstrual syndrome. A method according to claim 1 or claim 2, wherein the disease or condition isassociated with wasting.

6. A method according to claim 1 or claim 2, wherein the disease or condition isassociated with anxiety.

7. A method according to claim 1 or claim 2, wherein the disease or condition isassociated with adenosine- associated pathologies.

8. A method according to any one of claims 1 to 7, wherein the composition comprises folinic acid.

9. A method according to any one of claims 1 to 7, wherein the composition comprises a physiologically acceptable salt of folinic acid or mixtures thereof. A method according to any one of claims 1 to 9, wherein the composition is administered by a systemic or topical route. l s3/-/"I i -17

11. A method according to claim 10, wherein the systemic or tooical route is selected from the group consisting of oral, inhalable, topical, parenteral, and transdermal.

12. A method according to claim 11, wherein the route of administration is selected from the group consisting of buccal, sublingual, deral-, intraocular, subcutaneous, intradeimal, int-ramuscu.ar, intravenlous and intraarticular.

13. A method according to claim 11 or claim 12, wherein the composition is administ-ered as an oral formulation selected from the group consisting of capsules, cachets, lozenges, tablets, pov.-der, granules, solutions, susp~ensions and emulsions. ILI. A method according to claim 13, wherein the solutions and susnensions are selecte6 -from the group consisting of aqueous and. non-aqueous liquid solutions and suspensions. :15. A method according to claim 13, wherein the emulsions are selected from the group consistinlg of oil-in- water and water-in-oil emulsions.

16. A method according to claim 11 or claim 12, wherein the comoosition~ is ad: .4 nsered as a buccal or sub- lingual formulation selected from the group consisting of lozenges further comprising a flavoring agent selected from the group consisting of sucrose, acacia and tragacantilh; and castilles Further comroising an iinerti base selected from the group consisting of gelat n glycerin, sucrose and acacia.

17. A method accordi-ng to claim 13, wherein the oral formulation further como-1-ses an enteric coating- is. A method. accordin~g to claim 11, wherein the comDosit'ion is administered as a narenteral formulation selected from the group consisting of injectable solutions or suspensions, which rtay further comprise antioxidants, buffers, bacteriostatic agents and solutes which render the isS, solution or suspension isotonic wit~h the blood of any intended recipient i9. A method accorin.fg to claim 138, wherein the solutions and suspensions are selected from the group consisting of sterile actueous and non-aqueous injeccion and suspensions, which may Further comp-rlse suspending agents and thickening agentI s. A method according to claim 10, in unit-dose -form.

21. A method according to claim 10, wherein the formulation is in bulk or multi-dose form.

22. A met'-od according to claim 18, wherein the parenteral formulation is orovi'ded in bulk or multi-dose *form selected from the group) con Si sting off sealed amuoules and vials.

23. A method according to claim 10, wherein the -frmulation is freeze-dried or lyophilized; and the method further comocrises adding a sterile liquid carrier selected from the group consisting of saline and water prior to use. 20 24. A method according to claim 11, wherein tne comoposit-ion is administered as a tooical formulation selected from the group cons isti ng of ointments, creams, lotions, loastes, gels, sprays, aerosols and oils; ahich may furthner co'norise a carrier selected f 'rom the group consisting of vaseline, lanoline, polyethylene glycols, alcohols and trans-der-mai enhancers. A metnod according to claim I1, wherein the compoosition is administered as a transderrnal formulation in the form of a patch.

26. A mtechod according to claim 25, wherein the composition is administered as a Pa_-enieral form~ulation selected 'rom the group cons iSt4 ng of- ionropho-retic device comprising a solution or sus-penslion of the agent, which may urther comprise a buffer.

27. A method according to claim 11, wherein the composition is administered as an inhalable formulation. 19 2S, A method according to claim 27, wherein the inh-alable forMulation is an aerosol comprising liaou-o. or solid particles of the agent, ari which may further ccmorise an, agent selected From the g-roup consisting of preservatives, antn.oxidants, flavo'ring agenzs, volatile oils, buffering agents, disoersanzs and. surfactants.

29. A miethod according to claim I, loinereinthagn -is administered in an amount of about I to 1,000 mg/kg~ body Wt.

30. A maethod according to claim 1, wherein the agent is adininszered Jn an amoun't of about 5 to 500 mg/kg body Wt.

31. a method according to claime 1, wherein the comoositc.on furtner comorises an age-it selected from the group consisting of -preservatives, antioxidants, flavoring agents, volatile oils, bufFer~ng agents, dispersanits and surtactants

32. A method according to claim 1, tshere-in the ohvsiologically accezotable salt is selected from the group consisrting of alaiemetal, alkaline earth salts and organic salts.

33. method accordi-no Lo claim 51, wherein the physiologiJally acceptable salts are selected f-'rom the group consisting of sodium, potassiumi, calcium arid carboxvlic acid salts.

34. A method according to claim -wherein the subject is a human. A method according to clim 1, wherein the subject is an animal.

36. A method according to claim I, wherein -Which is a oronhvlactic method.

37. A method according to c-laim 1, which is a theraoeut ic method.

38. An in vivo method of Preventiing or treating a disorder or condition associated with low levels of adenosine or adenosine depletion, comprising applying to the subject an agenTt disclosed in the method according to 20 claim 1, wherein the amount of the agent administered to the subject is effective to increase adenosine levels and, thereby, revent or treat the disorder or condition.

39. A method according to claim 38, wherein the disorder or condition is selected from the group consisting of disorders or conditions of tne hleart, liver, JIUnQ (S) and brain. -P method -accevding to claim 39, whnerein t:he tissue is the subject.'s lung 1 C 41. A method according to claim 34, wherein Uthe -,issue is the subject-'s brain. 4.A method accordinci to claim 38, wherein. the tissue is the subject-'s heart- 4-3. A meth-od according to claim 4-2, wherein the tissue is the subject's liver.

44. A method according to claim 43, wherein th pnys ioloc i cally acceptable crircomprises a -onarmaceuticallv acceotable car-rier. A -method according to claim 44, wherein the connositio-~ is administered concurrenitly with other agents.

46- A method according to claim 1, wherein the agent 4s adm :-itered in an~ an- s--convulsant, anzi-anxiolvtic and neuro~rotective acents.

47. A metnod. according to claim 46, wherein the low Levels of adenosine or adenosine dep~letion are associated with convulsions.

48- A4 method according to claim 46, wherein the low levels of adenosine or adenosine dep:letion is associated with a mood condition. 4,'9 A method accord-ipg to claim 46, wherein the low levels of adenosine or adenosine deplet ion is associated with a neurologic condition. A composition, comprising an agent selected from the group consisting of folinic acid, physiologically acceptable salts thereof and mixtures thereof.

51. A composition according to claim 50, further compr-ising a carrier.

52. A composition according to claim 50, wherein the carr ier is selected from th'e group- consisting of solid anrd liauid carriers.

53. A poharmaceutical comoos-ition, comprising A composition according to claim 50, w.:'erein the carrier comprises a olnarmaceuticaliy acceptable carrier.

54. A compoos-ition according to claim 50, further comprising agent selectec. from the group consist-in of antioxidants, flavoring aciens, volatile oils, buffering agents, disuersants, sl-rzaczants, orozzellants and oreservatives. 5 Aformulation, comorlsino A, comnposition according to claim 53, w-hich i-,s a svstemic or t opical formulation.

56. A formui.tion accordai-ng to clm 55, -wherein the svstzemiic or tozpical f ormulation is selected from the group consisting of oral, irhalable,toia, aenrlad transderrm.al f or-mulations

57. A formiulation according to claim 56, wherein the formulation is selected f'rom.- the group consisting of buccal, sublinpual, dermal, :ntraocular, subcutaneous, intrade=mal, intrarmuscular, intr-avenous and intraarticular f or-mulat "I-ons

58. A formulation accoring to claim 57, which is an oral formulation select-ed from the group consisting of capsules. cachets, lozenges, tablets, powder, granules, solutions, suspensions and emulsions.

59. -A formulation according to claim 58, wherein the solutions and suspenslions are selected fromn the group consisting of acoueous and no-n-acueous, licuid solutions and suspensionas. A formulation according to claim 58, whereinr tine emulsions are selected from the group consisting of o--n- water anda water-in-oil em~ulsions. 6i-. formulatio-n according to claim 56, -which is is a buccal or sub-lingual formulation selected from the group consisting of 22 loegsfUrther coj.or s~flg a __avor2-ng agent selected from hegr-ouD Consisting of: sucrose, acacia and tragacanth; and oastl!C :utne copriS :nO an 4nert base selected from ~egroulo cor.Sisjlgo gelatin, 9lcrn Ur5 n acacia.

62. A~ oral formul at ion acc-OyC~n to the oral formul ation fur-ther coMOrJ5e an etrCcaig G3. A om2a~nacr2n oclaim 56, hihis a earnteaL or~~atflselected from rthe group coTISISti f ijetabe sluIzons or susp nSoS whh may furte comprise cx idanzs, burtfers, baczteriostatic agents and solutes wh-ichn ren-der the so, urion or suspensionl isotOZ11c with tlhe blood of any 4nzencded recrplent C4 A ormlt~ accordiz-g to claimL 63, lahere1Ln the slutinS ad sup~ -sOS are selected from the group coasisting of srerl.Le aqueous and ~nnaqueous inJection solutions and susoensiofls, wn___h ray Further comprise susoending agents and tnickening agents. A om Lon according tclaim 55, inl single or multiunit-dose f crT-m.

66- A forrulatlon according to claim 55, in bulk. Aeareneral ormulation accorditic to claim 0o, ~nbulk or multi-dose form selecte from the group consisting of sealed awmpoules and vials. 6S, A formulation according t o cla i-m 5 5, whiJch i s freeze-dried or ivooh~ized.

69. A formulation according tclaim 56, which, is a tooJ-cal formulation selected from the group corssSting of ointments, creams, lotions,. Pastes, gels sprays, aerosols and oils; w. hich =,av further conm-rise a carrier selected from the groupcon 0 sistinlg o-f vaseline, lanolinle, polyethyleneC glycols, alcohols and trans-Qer-mal enhancers. A for=itlatioii according to claim 56, -which is a transdermal fformulaFCiof comprised in a patch. -7 A formulation according to claim 70, w-hiclr is a oarenteral formulation in the formn of a solution or 13")1199 23 suspoension of the agent, wbhn may further comprise a buf"fer.

72. A method according to claim 36, which is an innalable -formnulat ion.

73. rnethod according tc. claimt 72, wher ein Lhe inhalable -formulation is an aerosol comprisIng liquid or solid Darticles of the agenz, arndwic may further comoPrise an agenC Selected rz the group consisting of oreservatives, antiox-ats, 'felvorinc] agen-_s, volatile oils, buflffering -agents, cas-ersanzz and surfactanzs.

74. A formulaclon according to claim 55, wherein the carrier comnrises a nvYdroh.obic carrier. A formulaarion according r-o claim 74. orovided in a composite in a separatre container with a delivery device. 13 76. A co-rnosiu e according r-o claimt 75, wn-erein the delivery device comor-4ses an in'aiazo_ which delivers individual ore-merered doses ot 'e f'ormulationi. *77. A composite according to claim. 76, wnerein the ~nalator comprises a nrebul'zer or isfltr n ute compr iig a nilercable or onenable capsule or cartridge wi th solid particles of the co-mposition. *78. A composite according to claini 75, -vh'erein the deiver devce cmprses a pressurized inhaler, and the tozrmato COIV.,ri ses a SUSoenSjQ_ or solution inan aqueous or -non-aareous ±-i-uid or an oJ1 i-n-water or oa-,er- rn-onl

79- A formulatio-n according to claim 74, provided i.n a capsule. A oarenteral formulation according to claim 71, comprising an injectable fo--nulation. Dated this 13th day of Jan~uary 1999 EAST C-AROLINTA UNIVERSITY B y th-eir Patent Attorneys GRIFFITH HAkCK Fellows institute of Patent and Trade Mlark Attorneys of Australia