United States Patent ‘US006630507B1 2) (0 Patent No: US 6,630,507 BL Hampson et al. (as) Date of Patent: Oct. 7, 2003 (58) CANNABINOIDS AS ANTIOXIDANTS AND OTTER PUBLICATIONS. NEUROPROTECTANTS: Windholz et al, The Merck Index, Tenth Edition (1983) p. Dat abstaet No. 733 (75) Inventors: Aidan J. Hampson, Irvine, CA (US); ‘Mechoulam et al ‘Total Synthesis of di—A'—Tetra- Zillos Arelrd, Reckile, MD (US. fydrocanabinl, ie Active Consent of Hash” ure Mauro Gelmal Betbeste, MD 2 the morc Chemical Sac, ST4S279-3295 7 (1965). (9) sigs The Ut Stain of Americ ay Mel, Cha if Hai Ay represented bythe Department Ottrsea et aly “The Crystal and Molecular Stueture of a Pa Cannabidiol,” Acta Chem. Seand. B 32, 9807-812 (1977) ne Cunha et al. “Chronic Administration of Cannabidiol 10 (+) Notice: Subject any disclaimer, the term ofthis Healthy Volunteers and Epileptic Patients!” Pharmacology, tent is extended or adjusted under 35 21:175-185 (1980) peat eroded of al Conse et sy “Acute and Chronic Anvepiepic Drug Y Eee in Andogenic Seimre-Stacepuble Hates Expon (21) Appl Nos ovjo7soas ‘mena Nesolgy, Academe Press ne 1620-037 (1080) ‘Tarkan tal, "Eectophysiobgic Pipers of he Cat 7 tubinlds J. Clin. Pharmacol, 2144564638 (22) PCT Filet: Apr 21,1999 Ca spon ad Avelpepic Ee of Cane ta hn Pharmacol A878 8) (60) PCTNos — PCT/USI9ION769 fel als “The Canmabincis as Potenal Ape tae! Cl Pharmac 37S 4485 (81), S31 OD), Constoe etal, “Antiepleptic Potential of Cannabidiol Anal- ©, (0 Dae: Feb. 2, 2001 05°F. Clin: Pharmacol, 34388-4368 (1981) (67) PCE Pub. Nos WO99/S3917 (Lit continue on wet page) PCT Pb, Date: Ot. 2, 1999 Primary Exaniner—Kevin E. Wetngson (9) Attorney, Agou,or Frm Klan Spartan, LLP Related US. Application Data pas fcr (60) troviiontappcation Ne 6018258) fed on Ape 21, 1855 anand opcaion No, 60095993, eden Cannabinoids have been found 10 have antioxidant ‘Auge 10, 1998, properties, unrelated to NMDA receptor antagonism. This te’ foun prope takes canabioide ws inte ea 6) mar AGIK 31238 ent and propfusis of wide vary ef euaaion aso (3 US. SLAMS Sted deeses suc as echomic, agesled infamcsiory ($8) Fleld of Search 514488 and autoimmune deus, The camabinois ave found 10 have parca pplication as neuoprlecant fo example 6 References Cited in iting neurological damoge following chemi inal suc as stoke an trauma, ofa te eaten of sured: US, PATENT DOCUMENTS ‘generative diseases, such 1s Alzheimer’s discase, Parkin- 230400 A 121042 Adons searag Son's dscase an HIV" dementia, Nonpayehoactive ae ATG Mia SS So sah ms pry sn "Stoo Starts SOuBLL tapes fo tse because ey avoid onic lhl encoun a imped StSAS4 ed wih poychontive conmbinndsat high doses wel in ‘A795 Meclam taln: S671 te met the presen invention, A partly dslosed a Py ee 514315 class of cannabinoids useful as neuroprotective antioxidants Ssizam Ch asf RAMS stomata) whee the group independ selected 5538093 A ‘Mechoulam ct al. ows $14/454 0m the group consisting of H, CH, and COCHs, Sassi A Nctoulm cal Stgase ie A Nattey Camere al stares ® ois388 Bt ea Paden cals cers IAS iw FOREIGN PATENT DOCUMENTS we 7518 AL soo1 ra Stas al 12ions ra sseast at “saoes fe ssnses at Wo woosenat AL Ee : wo wood12667 AL nde a Wo Wovelziss At Wo Wovetasoo At Wo Wome At wo 230 26 Claims, 7 Drawing SheetsUS 6,630,507 BL Page 2 (OTHER PUBLICATIONS, Colasaati etal, “Ocular Hypotension, Ocular Toxicity ad Neurotoxicity in Response to Marihuana Extract and Can- mabidiol,” “Gen Pharm, Pergamon Press Lid, 15(6):479-484 (1988) at lana Pras, Oa Toy a ter Administration of Cannabinol or C nabigerl,” Bap. Fye Res, Academic Pes ne, 39:23 (1984) Voltecta., "Cannabinoids Block Release of Serotonin from Platelets Induced by Plasma frm Migraine Patients,” Int. J Clin. Pharm. Res Bioscience Ediprint Ine 4243-246, 9859). ‘Agutell ct al, “Pharmacokinetics and Metabolism of A Tetrahydrocannabinol and Other “Cannabinoids with Emphasis on Man®,” Pharmacological Reviews, 38(1):21—48 (1986), Karler et sl, “Different Cansabinoids Exhibit Diferent Pharmacological and Toxicological Properties,"NIDA Rex. Monogr, 79:96-107 (1987). ‘Samara eta, “Pharmacokinetics of Cannabidiol in Dogs,” Drug Metabolism and Disposition, 16(3):469-472 (1988) ‘Choi, “Glutamate Neurotoxicity and Diseases of the Net vous System,” Newran, Cell Press, 1:623-634 (1988), Eshhar ot al, “Neuroprotective and Antioxidant Activities of HU-211, A Novel NMDA Recepior Antagonist,” Journal of Pharmacology, 283:19-29 (1995). ‘Skaper et al, “The ALIAmiée Palmitoylethanolamide and Cannabinoils, but ot Anandamide, ate Protective in & Delayed Posigtamate Paradigm of Exeitotoxic Death in Cerebellar Granule Neurons,” Neurobiology. Proe. Nal ‘Acad. Sci. USA, 93:3984-3989 (1996). ‘Alonso et al. "Simple Syathesis of 5-Substituied Resor. nols: A Revisited Family of Interesting Bioactive Mol- ecules,” J Org Chem, American Chemical Societ 20,1421 (997, bes et al. “A Simple Synthesis of the Natural 2,5-D Styli! Fee, Radel Seaenser Anion Resorsation,” Syrhetic Communications, Marcel Dekker, Inc, 27(21):3769-3778 (1997). 1 al, “Oxidative Stress in Closed-Head Inju Journal of Cerebral Blood Flow and Merabo- lism, Lippincott-Raven Publishers, 17(10):1007-1019 1999). Zarier et al, “DimethylheptylTHC-11 OIC Acid," Avthri- fis & Rheumasim, $1(1): 163-170 (1998) Hampson et al, "Dual Elects of Anandamide on NMD, Receplor-Mediated Responses. and Neurotransmission Journal of Newrochemisiry, Lippincott-Raven Publishers, 70(2)-671-676 (1998). Hampson et al, “Cannabidiol and (-)4°-tetrahydrocanns biono ace Neuroprotective Antioxidants,” Medical Sciences, Proc, Natl, Acad. Sei, USA, 8268-8273 (1998), * cited by examinerU.S. Patent Oct. 7, 2003 Sheet 1 of 7 US 6,630,507 BL = Le = ac Z > B “3 a8 a & 5 2 2 ey . Cy 2 y _ 9 s 2 a a go 3 = : ot = eet S = o 5 3 4 2 E 5 2 . t e a a esea[el HAT WD %US 6,630,507 BL Sheet 2 of 7 et. 7, 2003 US. Patent FIG. 2 % |S | EARNS UV t 5 eM CBD 1oOpM SpM THC GhUS 6,630,507 BL Sheet 3 of 7 et. 7, 2003 US. Patent Ww) [ud] SHOA Or Sh Os SO fe Al i tn ' \ r : 1 0 {0-76 $81 =OHL™)a “ry wit F1l=a9o "0a wit #8i=1Ha"0d Lez B wear § e é 8 § 8 ve 6s = 3 3 s sl wre 001 POM e OM (wn) quoun,U.S. Patent Oct. 7, 2003 Sheet 4 of 7 US 6,630,507 BL FIG.5 B A 100} 100 ge g : 2 a | Bx ge 3 as as ot s $5 60 55 50 48 49 jpouM CBD BET AscorbateTocopherot Glu feannabidiot} (M)U.S. Patent Oct. 7, 2003 Sheet 5 of 7 US 6,630,507 BL 6 [Linoleic acid (substrate)] (M) a 2 a o ‘A dd WA | Aq uomnqryut % oO = + g oo 2 : r | [Cannabidiol] (M) 2 s 8 (utu p€Z) aBueyo sqe [0 Jo %U.S. Patent Oct. 7, 2003 Sheet 6 of 7 US 6,630,507 BL 7 I~ z — 2 = el 08 "& \ & 7 2 s 7 (uu 9¢7) z axe yead [19 9% a z Q 2 i? = 2 | 28 _ é < 3 a 28 s 3 © = (wu 9¢z) 1 8 are 3yead [199 9% S ma — ° (wu 9¢z) vvore 3v0d 19 %U.S. Patent Oct. 7, 2003 Sheet 7 of 7 US 6,630,507 BL During & Post Post ischemia Time of 12-HETE application (0.5 g/ml) FIG. 8 During ischemia Ctrl uoronpord HT [9 Jo %US 6,630,507 BL 1 CANNABINOIDS AS ANTIOXIDANTS AND NEUROPROTECTANTS. “This application isa 371 of PCT/US99/08769 fled Apr. 21, 1999, which claims benefit of No. 082,589 fled Apr. 21, 1998, which eluims benefit of No, 60095,095 fled Aug. 10, 1998, FIELD OF THE INVENTION ‘The present invention concerns pharmaceutical com- pounds’ and compositions that are useful as tissue protectant, such as neuroprofectans and eardioprotectants, ‘The compounds and compositions may be used, for example, in the teatment of acute ischemic neurological insulis of chronic neurodegenerative diseases, BACKGROUND OF THE INVENTION Permanent injury to the central nervous system (CNS) ‘occurs in variety of medical conditions, and has been the Subject of intense scientific scritiny in recent years. I is ‘known that the brain has high metabolic requirements, and that it ean suffer permanent neurologic damage if deprived ‘of sulicient oxygen (hypoxia) for even afew minutes. Inthe absence of oxygen (anoxia), mitochondrial production of AIP cannot meet the metabolic requirements of the brain, 3 and dssue damage occurs. This process is exacerbated by neuroma release of the neurotransmiler glutamate, which stimulates NMDA_(N-methyl-D-aspartate), AMPA, (ce-amino-3-hydroxy-5-methy-4-soxazole propionate) and Kanate receptors. Activation of these receptors initiates calcium infltx ito the noutons, and prodiction of reactive ‘oxygen species, which are potent toxins that damage impor- fant cellular siractures such ts membranes, DNA and enzymes, ‘The brain has many redundant blood supplies, which means that iis tiseve is seldom completly deprived of ‘oxygen, even during acute ischomie oventsesusedby thom- ‘hoombolic events oF trauma, A combination of the injury of hypoxia with the added insult of glutamate toxicity therefore believed to be ultimately responsible for celular leah, Hence ifthe additive insult of glutamate toxicity can bo alleviated, neurological damage could alsa be lessened ‘Antisonidanis ad ani talory agents have been pro= posed t reduce damage, but they often have poor acces 10 Struetures such as the brain (which are protected by the ‘ood brain barcer), Given the importance of the NMDA, AMPA and kainate receptors in the mechanism of injury, research efforts have focused om using antagonists to these receptors to interfere with the receptor mediated calcium influx that ultimately leads to celular death and tissue necrosis, In vio studies using cultured neurons. have demonstrated that glutamate receptor antagonists reduce neurotoxieity, but NMDA and ‘AMPA\kainate receptor antagonists have diferent effets. ‘Antagonists to NMDAr prevent neurotoxicity if present luring the glutamate expesure period, but ae less elective if added alter glutamate is removed. In contrast, AMPA) kkaate receptor antagonists are not as effective as NMDA, Antagonists during the glutamate exposure period, but are more effective following glutamate exposure. Some ofthe research on these antagonists has focused on ‘cannabinoids, 2 subsel of which have been found to be NMDA receptor antagonists. US. Pat. No. 5,538,093 (3S, «4S.delta.6-ttrahydrocannabinol-T-oie acids), US. Pat, No, 5,521,215 (sterospecilic (+) THC enantiomers), and US, Pat, No. 5,284,867 (limethytheptyl_ benzopyrans) have reported tha those cannabinoids ate effective NMDA recep tor blockers. US. Pat. No. 5,134,295 discloses thatthe 1 climethylheptyl (DMA) homolog. of [SRSR}-T-hyeoxy- 2 A°TIIC (known a6 HU-210) isa supepotent cannabinoid receptor agonist with esnnabinomimetisatvity 0 o¢ders ‘of magnitie greater han the natural 4° THC, Te HU-210 Sintra cli sre ie fe ing itt, headache, and hypotension. J. Pharmacol Sa or 433-1457 (1971). Subjects who recived th sa. thetic cannabinoid witha dimethylbepty group experienced marked psychomotor eetacation, ad were: vowing o¢ incapable of assuming an eret positon In contrast to HU-210, the (-)GRAR) THC-DMIL enan- tomer (knowo as HU) displays foe ality 10 the ‘cannabinoid receptors, but retains NMDA receptor ano nist neuroprotective wf ‘THC (etrahydrocannabinol) is another of the eannat inoids that has been shown to be neuroprotective in cell ‘cultures, but this protection was believed to be mediated by interaction at the cannabinoid receptor, and so would be accompanied by undesired psychotropic sie effects. oy on * cHgCH CH Abbough it hss been unclear whether cannsbimimetic activity plays a role in neuroprotection against glutamate induced neurological injury, the teaching in this field has ‘leary beea that a cannabinoid must at least be an antagonist atthe NMDA receplor ta have neuroprotective elle, Hence cannabidiol 2-[3-methyl-6-(1-methylethenyl)~ ‘yeloliexen-L-yl}5-pentyl-t,-benzenediol or CBD), a can habinoid devoid of psychoactive elfeet (Pharm. Rev 38:21-43, 1986), has not been considered useful a8 neu- roprotectant, Cannabiiol has been studied as an antepilep- tie (Carin etal J. Clin. Pharmacol. 21:M178-427S, 1981; Karler et al, J. Clin. Pharmacol. 21:437S-4488, 1981, Constoe et al, J. Clin Phannacol. 21:4285-436S, 1981), and bis been found wo lower inioculae pressure (Colasanti etal, Exp, Eve Res, 39:251-259, 1984 and Gen, Pharmac, 15:479-484, 1984). Combi RD)US 6,630,507 BL 3 No signs of toxicity or serious side effects have been ‘observed following chronic administration of eannabidiol to healthy volunteers (Cunba et al., Pharmacology 21:175-185, 1980), even in large acute doses of 700 mgiday (Consive et al, Pharmacol, Biochem. Behav: 40:701-708, 1991) but eanmabidiol is inactive at the NMDA receptor Hence in spite ofits potential use in treating glaucoma and seinites, cannabiio! has not been considered a neuropro- Tective agent that could be used to prevent glutamate induced damage in the central nervous system, SUMMARY OF THE INVENTION tis an object ofthis invention o provide a new clas of antioxidant deugs, that have particular application as, neuraprotectants, although they are generally useful ia the ‘neatmeat of many oxidation associated diseases ‘Yet another object of the invention isto provide a subset ‘of suc drugs that can be substantially free of pyehoactive fr psychotoxie effets, ae substantially non-loxie even at very high doses, and have good tissue penetration, for ‘example crossing the blood brain barrier. tas surprisingly beon found that eannabidiol and other cannabinoids can function as neuroprotectants, eventhough they lack NMDA recepior antagonist activity. Tisdliscovery ‘was made possible because ofthe inventor's recognition of 4 previously unanticipated antioxidant property of the can nabinoids in general (and cannabidiol in particule) that functions completely independently of antagonism at the NMDA, AMPA and kainate receptors. Hence tbe present invention includes methods of preventing or treating dis: ceases caused by oxidative sires, sich as neuronal hypoxia, by administering « prophylactic or therapeutically effective Amount of a cannabinoid to a subject who has a disease ‘caused by oxidative sess ‘The cannabinoid may be a cannabinoid other than THC, HU-210, or other potent cannabinoid receptor agonists. The cannabinoid may also be other than HU-211 or any other NMDA receplor antagonist that has previously been reported. A potent cannabinoid receptor agonist i one thst has an EC atthe cannabinoid receptor of 50 nM o# less but in more particular embodiments 190 nM of 250 nM oF les. In disclosed embodiments the cannabinoid is not ‘psychoactive, and is not psychotoxic even at high doses. In some particularly disclosed embodiments, the cannabinoid is selested from the group: where A is aryl, and particularly Bs re s 4 but not pinene such as SB and the R,-R, groups are cach independently selected from the groups of hydeogen, lower substituted or unsubstituted alkyl, substituted or unsubstituted carboxyl, substtued or unsubstituted alkoxy, substituted or unsubsiituted alcobol, and substituted or unsubstituied ethers, and R,-R, are H of methyl. In particular embodiments, there are no nitrogens in the rings, andor no amino substitutions on the rings. In other embodiments, the cannabinoid is one of the followings where there ca be 0 10 3 double bonds on the A ring, as indicated by the optional double bonds indicated by dashed Tines on the A ring, The C ring is aromatic, and the B ring ‘ean be a pyran, Particular embedimenis are dibenzo pysans and cyclohexenyl beuzenediols, Particular embodiments of the eunnabinoids of the present invention may aso be highly lipid soluble, and in particular embodiments can be diUS 6,630,507 BL 5 solved in an aqueous solution only sparingly (For example 10 mgiml or less). The octanol water partition ratio. st neuteal pH in useful embadiments is S000 or greater, for ‘example 6008 or greater. This high iid solubility enhances. penetration of the drug into the CNS, as reflected by its Volume of distribution (V,) of 15 Likg or more, for example 23.5 Likg, 7 Likg, or ideally 10 Lkg or more, for example at least 20°Lkg, Particular embodiments may also be highly ‘water soluble derivatives that are able te penetrate the CNS, For example earboxyl derivatives, Req afe independently selected from the group of Hy subslitited or unsubstituted alkyl especially lower alkyl, for ‘cxample unsubstituted C.-C, alkyl, hydroxyl, alkoxy, espe ially lower alkoxy such as methoxy or ethoxs, substituted (or unsubstituted alcobol, and unsubstitued or substituted ‘cuboxyl for example COOH or COCH,, In other embodi- ments Ry, can also be subsiituted oF unsubstituted amino, and halogen The cannabinoid has substantially no binding to the NMDAr (for example an ICzo greater than or equal to 5 4M ‘10 M), has substantially no psychoscive activity medi ated by the cannabinoid receptor (for example an IC, atthe cannabinoid receptor of greater than or equal 10 300 nf, for example greater than 1 jf and « K, greater than 250 oM, expecially 500-1000 oM, for example greater han 1000, 1M), snd antioxidant activity, as demonstratable by the Fenton reaction of eyeie voltameity nother particular embodiments, the eannabinoids are one of the followings Re Re, where Ryo is substituted oF unsubstituted alkyl, such as. lover allyl (for example methy), lower alcohol (such as methyl alcohol) or carboxyl (such as eatboxslic acid) and ‘oxygen (as in =O); Ray is hydrogen or hydzoxy; Ray is hydeogen, hydroxy, of methoxy; Res is hydrogen or hydroxy; Ry i hydrogen or hydroxy; Ris hydrogen or hydroxy; Ra, is hydrogen or bydroxy, and, is substituted ‘or unsubstituted alkyl (for example n-methylalky), substi tuted or unsubstituted alcohol, o¢ substituted oF uasubsti- tuted carboxy, 6 In yet other embodiments of the invention, the cannab- inside are 1, 0% Seats ne ‘wherein numbering conventions for exch of the ring pos tions are shown, and Rey, Reg anal Ray are independently selected from the group consisting of H, unsubstitted lower alkyl such a8 CH,, and carboxyl such as COCH,, Pacticulae ‘examples of nonpaychoactive cannabinoids that fall within this definition are eaanabidiol and tnd other sisuctural analogs of eannsidiol In more particular embodiments, the cannabinoid is used to prevent or ica an ischemic or neurodegenerative disease in the central nervous system ofa subject, by administering to the subject a therapeutically effective amount of @ ean- habinoid 10 protec against oxidative injury 1 the central nervous system, The cannabinoid may be any of the con pounds set forth above, oF more specially ‘wherein Rey, Rag tnd Ray are independently selected from the group conssting of H, lower alkyl such as CH, and carboxyl such ss COCH,, and particularly wherein 2) RyeRayeRacoll by ReyeRoe: RogeCH, ©) ReyeRagaCHly; Regal 0) ReyeRageCOCH; Rogol coc, then the compound is eannabidiol '=Reyatl and RuxCHs, the compound is CBD mopomeiiyl etber. When ReyeRoy=CH and) RveHl, the ‘compourel is CBD dimethyl ether When Ra =Raa=COCH, Hl the compound is CBD dliaeetate. Wien R.y=FL =RegeCOCH the compound is CBD monoacelat. ‘The ‘ischemic or neurodegenetative disease may be, forUS 6,630,507 BL 7 le, an ischemic infarct, Alzheimer's disease, Parkia- son's disease, Down's syndrome, human immunadefiiency virus (HIV) dementia, myocardial infareton, or treatment and prevention of intraoperative or perioperative hypoxic insults that can leave persistent neurological deficits follow ing open heart surgery requiring heart/lung_ bypass machines, steh as coronary artery bypass gralis (CABG). ‘The invention also includes an assay for sclecting. cannabinoid to use in tating a neurological disease by ‘determining whether the cannabinoid is an antioxidant, ‘Once it has been determined that the cannabinoid is an antioxidant, an antioxidant elfective amount of the eannab- ind is administered to teat the neurological disease, sch as vascular ischemic event inthe central nervous system, for example ihe type caused by a neurovascular thromboem= holism. Similarly, the method of the present invention includes determining whether a disease is caused by oxida- live tres, and ifthe disease is caused by oxidative stress, ‘administering the cannsbinoid in a therapeutically effective Antioxidant amount The invention aso inchues identifying and administering antioxidant and. ncuroprotective compounds (such as annabidiol) which selectively inhibit the enzyme activity of both S- and 15-lipoxygentse more than the enzyme activity of 1 low NMDA antagonist activity and low cannabinoid recep. {or ativity, Assays for seleting compounds with the desired clfect on Hpoxygenase enzymes, aid methods for using. idomtitied compounds to eat neurological or ischemic dis- eases are also provided. Such diseases may include a vas culae ischemic event in the central nervous system, for example a thromboembolism in the brn, or a vascular ischemic even in the myocardium. Useful administation of the compounds involves administration both during. and after an Ischemic ijury ‘These and other objects of the invention will be under stood more clearly by reference to the folowing detailed ‘escription and drawings BRIEF DESCRIPTION OF THE FIGURES FIG. 1A js a graph showing NMDA induovd cellular damage in a neuron (as measured by LDH release) in cells That were exposed to glutamate for 10 minutes, which ‘demonstrates that increesing concentrations of eannabidiol in the cell culture proteets against cellular damage. FIG. 1B is a graph similar to FIG. 1A, but showing that AMPAlkainate receptor mediated damage (induced by slutamate and the AMPA‘Kainate receptor potentiating agents eyclothiazide or concanavalin A) is alsa reduced in a concentration dependent manner by the pesence of canna- bidiol in the culture medium, FIG. 2A is a bar graph showing cellular damage (as measured by LDH release) in the presence of glutamate sone (100 uM Glu), sod inthe presence of glutamate and 5 41M cannabidiol (CBD) or 5 iM THC, and demonstrates that, ‘CBD and THC were similarly protective, FIG. 2B isa bar gsaph similar to FIG.2A, but showing the cellular damage assessed in the presence ofthe eannabinoid receptor antagonist SR 141716A (SR), which was not found toalter the neuroprotective eeet of CBD (5 4M) or THES HM), indicating the elect isnot atypical cannabinoid ellect, mediated by the eannabinoid receptor. FIG. 32 graph showing the reduction oxidation poten- tials determined by eyelic voltametcy for some natural and synthetic cannabinoid, the antioxidant BHT, and the non- ‘cnnabinoid anandamide (aachidonylethanolamice) which lipoxygenase, In addition, such compounds posses. 25 8 ‘sa ligand forthe cannabinoid receptor. The voltage at whet initial peaks occur is an indication of antioxidant activity FIG. 4 is a graph that demonstrates the antioxidant properties of BHT, CBD axl THC, by posting the fuores- ‘cence of a fuoresceat dye against concenteations of these substances, where declining fluorescence i an indication of ‘greater antioxidant activity. FIG. 5A is graph illustrating decreased -buty] peroxide induced toxicity (as measured by LDH release) in the presence of increasing concentrations of eannabidiol, dem- ‘onsirsting that cannabidiol is an effective antioxidant in living cells FIG. SB is a bar graph comparing the antioxidant activity ‘of several antioxidants against glutamate indeed toxicity in neurons, showing that CBD hs superior antioxidant activ- iy, FIG. 6A is a graph showing the effect of CBD (as measured by the change ia absorbance at 234 am) oa the ‘enzymatic setivty of two lipoxygenase enzymes, rabbit 15-LO and porcine 12-LO, which demonsrales that CHD inhibits 15-L0, but aot 12-LO enzyme. FIG. 6B isa graph demonstrating that inhibitory elect of CBD on 15-L0 is competitive, FIG. 7Ais a graph similar to FIG, 6A, but was performed in whole cals rather than purified enzyme preparations, and shows the effect of CBD (as measured by the change in absorbanoe at 236 am) on the enzymatic activity of 5-LO from cultured rat basophilic leukemia cells (RBL2H3), ‘which demonstrates that CBD inhibits 5-LO. FIG. 7B is a graph showing the effect of CBD (as measured by the change in absorbance at 236 am) oo the formation of 12-HETE (the product of 12-10) by human leukocytes (12-L0 type 1). FIG. 7C isa graph similar to FIG. 7B, sbowing the effect, ‘of CBD (as measured by the change in absorbance st 236 fam) on the formation of 12-HETE by human platelets (12-L0 type 2). FIG. 8 is a bar graph demonstrating that 12-HETE ean protect cortical neurons from NMDAF toxicity most effee- lively when administered during and post ischemia, DETAILED DESCRIPTION OF SOME SPECIFIC EMBODIMENTS, ‘This invention provides antioxidant compounds and ‘compositions, such as pharmaceutical compositions, that include cannabinoids that act as fee radical Seavengers for use in prophylaxis and treatment of disease The invention also includes methods for using the antioxidants in preven- tion and treatment of pathological conditions such as ischemia (tissue hypoxia), and in subjects who have been ‘exposed to oxidant inducing agents such as cancer ‘chemotherapy, toxins, radiation, of other sources of oxidae tive ses. The compositions and methods described herein are also used for preventing oxidative damage in trans- planted oxans, for ihibiting reoxygenation injury follow Ing reperfusion of ischemic tssues (for example in heart dlisease), and for any other condition that is mediated by ‘oxidative or fee radical mechanisms of injury. In particular ‘embodiments of the invention, the compounds and compo- Sition are used in the treatment of ischemic cardiovascular ‘nd neurovascular conditions, and neuradegenerative dis- ‘eases, However the present invention ean also be used as an ‘ntioxidant treatment in non-neurological diseases, Molecular oxygen is essential for aerobic organisms, ‘where it participates in many biochemical reactions includ