Methods of use for peroxynitrite decomposition catalysts, pharmaceutical compositions therefor

The present invention provides a method for the treatment of diseases by the decomposition of peroxynitrite, preferably decomposition to benign products, comprising the use of a complex which is a selected ligand structure providing a complexed metal such as Mn, Fe, Ni and V transition metals. The method of use, as well as novel pharmaceutical compositions therefor, are for the treatment of diseases advantageously affected by decomposition of peroxynitrite ed at a rate over the natural background rate of decay of peroxynitrite in humans suffering from the disease which comprises administration of an amount of a complex, in dosage unit form, which is effective for such acceleration of the decomposition of peroxynitrite .

TECHNICAL FIELD
 The present invention is for methods of use for the decomposition of
 peroxynitrite by metal complexes, novel pharmaceutical compositions, and
 methods of use therefor.
 Particularly, the present invention now provides a method for treating
 selected diseases comprising the decomposition of peroxynitrite with the
 use of a compound which is a metal complex. This decomposition preferably
 produces benign agents preventing formation of deleterious decomposition
 products such as oxygen radicals and which also further prevents
 inactivation of superoxide dismutase (SOD) by the presence of
 peroxynitrite. Therefore, the method of use for selected metal complexes
 of the present invention, as well as novel pharmaceutical compositions for
 such use is for the treatment of diseases advantageously affected by
 treatment comprising decomposition of peroxynitrite at a rate accelerated
 over a natural background rate of decay which comprises administration of
 an rate-accelerating effective amount of the metal complex in unit dosage
 form.
 In other words, the methods of treatment and novel compositions of this
 invention provide a twofold benefit in the treatment of diseases (1)
 accelerated rate of catalytic decomposition of peroxynitrite and (2)
 protection of SOD against inactivation by peroxynitrite.
 Thus, the present invention provides for a method of treatment of human
 diseases advantageously affected by such decomposition by protection from
 the deleterious effects resulting from the presence of peroxynitrite in
 the human body not heretofore known. In addition, since protection against
 SOD inactivation is provided, such decomposition offers protection against
 diseases associated with the overproduction of superoxide.
 These diseases include ischemic reperfusion injuries such as stroke, head
 trauma and myocardial ischemia, sepsis, chronic or acute inflammation
 (such as arthritis and inflammatory bowel disease and the like), adult
 respiratory distress syndrome, cancer, bronchopulmonary dysplasia, side
 effects from drug treatment of cancer, cardiovascular diseases, diabetes
 (not included for treatment by vanadium porphyrin complexes), multiple
 sclerosis, parkinson's disease, familial amyotrophic lateral sclerosis,
 and colitis and specific neuronal disorders, preferably ischemic
 reperfusion, inflammation, sepsis, multiple sclersis, parkinson's disease
 and stroke.
 BACKGROUND ART
 Nitric oxide (NO) is known for its dual physiological role as helpful
 messenger and harmful intermediate. Nitric oxide is shown to be generated
 in numerous cell types including macrophages, neutrophils, hepatocytes and
 endothelial cells. See Hibbs et al, Science, 1987,235,473-476; Rimele et
 al, J. Pharmacol. Exp. Ther., 1988, 245, 102-111; Curran et al, J. Exp.
 Med., 1989, 170, 1769-1774; and Plamer et al, Nature, 1987, 327, 524-526;
 respectively. The chemical reaction responsible for the production of NO
 is catalyzed by a class of enzymes referred to as nitric oxide synthases
 (NOS) which convert L-arginine to citrulline and NO. Forstermann et al,
 Biochemical Pharmacology, 1991,42, 1849-1857. While the role of NO as a
 signaling molecule in the stimulation of guanylate cyclase is well
 established, (Monocada et al, Pharmacological Reviews, 1991, 43, 109-142),
 the origins of its cytoxicity remained unclear.
 Recently a body of compelling evidence surfaced which teaches that NO by
 itself may not be responsible for cell damage (See Absts. of 1st Annual
 Mtg. of Oxygen Society, Nov. 12-4, 1993, Charleston, S.C., "Nitric Oxide
 Requires Superoxide to Exert Bactericidal Activity" by L. Brunnelli and J.
 S. Beckman). Instead a more reactive species, peroxynitrite, produced by
 the reaction of superoxide and NO, is found to play a role in the
 cytotoxicity observed with the over-production of NO. Peroxynitrite is
 known to decompose via a process which is first order in protons. The rate
 of proton catalyzed decomposition of peroxynitrite (hereinafter "the
 natural background rate of decay") is understood from its study over a
 variety of pH ranges (see; Keith et al. J Chem Soc (A), p.90, 1969). When
 the pH is 7.4 and the temperature is maintained at 37.degree. C., the
 observed rate for the decomposition of peroxynitrite is
 3.6.times.10.sup.-1 sec-i (see Beckman et al. Proc. Natl. Acad. Sci. USA
 Vol 87, pp1620-1624, 1990). Beckman shows that peroxynitrite decomposition
 generates a strong oxidant with reactivity similar to hydroxyl radical, as
 assessed by the oxidation of deoxyribose or dimethyl sulfoxide with the
 further suggestion that superoxide dismutase protects vascular tissue
 stimulated to produce superoxide and NO under pathological conditions by
 preventing the formation of peroxynitrite. See Beckman et al, "Apparent
 Hydroxyl Radical Production by Peroxynitrite: Implications for Endothelial
 Injury from Nitric Oxide and Superoxide" in Proc. Natl. Acad. Sci. USA,
 Vol. 87, pp 1629-1624, February 1990.
 Further, it is well established that peroxynitrite decomposes to give the
 hydroxyl radical and nitrogen dioxide, a potent nitrating agent. Both of
 these species are potent oxidants shown to react with lipid membrane and
 sulfhydryl moieties (See Radi et al "Peroxynitrite Oxidation of
 Sulfhydryls" in The Journal of Biological Chemistry, Vol. 266, No. 7 March
 5, pp 4244-4250, 1991).
 Hardy et al suggest the interaction of O.sub.2- with nitric oxide forms
 peroxynitrite or the protonation of O.sub.2- to form perhydroxyl radical
 is involved in the neutrophil-meditated killing of HAE cells (FASEB
 Meeting on Apr. 5-9, 1992 in Anaheim, Calif.) and further Hardy et al
 suggest a role for peroxynitrite in oxidative damage of human endothelial
 cells (Abstract in the "Experimental Biology" section of FASEB on Mar.
 28-Apr. 1, 1993 in New Orleans, La.).
 In other words, harmful products from peroxynitrite decomposition is
 specifically taught by many references.
 In addition, it has been shown that the reaction of peroxynitrite with Mn
 and Fe SOD results in inactivation of the enzyme (See also Radi et al,
 Arch. Biochem. Biophys., 1991, 288, 481-487). It is now known that
 peroxynitrite will also inactivate CuZn SOD.
 Thus, the effects of the decomposition of peroxynitrite; whether by the
 generation of damaging decomposition products or inactivation of SOD, in a
 wide variety of diseases are well documented.
 For example, a study assessing the deleterious effects of peroxynitrite on
 the rat colon is reported by Rachmilewitz et al in "Peroxynitrite-induced
 Rat Colitis: A New Model of Colonic Inflammation" from Gastroenterology
 105 (6) 1993.
 Beckman et al in PCT/US91/07894 (corresponding to U.S. Pat. No. 5,277,908)
 teach, specifically that peroxynitrite is formed by the reaction of
 superoxide (O.sub.2-) and nitric oxide in tissues subjected to ischemic,
 inflammatory or septic conditions. Beckman et al link SOD deficiencies and
 peroxynitrite to amyotrophic lateral sclerosis (ALS) in Nature, Vol 364,
 12 August 1993 and Hogg et al and Beckman et al., respectively, present a
 relationship between peroxynitrite and atherosclerosis in Biochemical
 Society Transactions, Vol. 21, received Dec. 22, 1992 and in "Extensive
 Nitration of Protein Tyrosines in Human Atherosclerosis Detected by
 Immunohistochemistry", Biol Chem. Hoppe-Sevler, Vol. 375, pp 81-88,
 February 1994. Further, the involvement of peroxynitrite in various
 disease states is found for lung diseases attributed to cigarette smoke,
 atherosclerosis, amyotrophic lateral sclerosis, cold-induced brain edema
 in Chem. Res. Toxicol., Vol. 5, No. 3, 1992 pp 425-431. See also
 "Cold-induced Brain Edema in Mice" in The Journal of Biological Chemistry,
 Vol.268, No. 21 Issue of July 25, pp 15394-15398, 1993.
 More recently a spinal neuron toxicity assay has been developed by Scherch
 et al to screen for drugs which block peroxynitrite toxicity. (23rd Annual
 Meeting of the Society for Neuroscience, Washington, D. D., Nov. 7-12,
 1993 and abstracted in Society for Neuroscience Abstracts 19 (1-3) 1993
 and Biosis 94:4951.
 Further, by preventing inactivation of SOD by reducing the presence of
 peroxynitrite the present invention also provides enhancement of known
 physiological benefits of superoxide dismutase in the treatment of
 diseases based on such benefits. In this regard SOD and its mimics have
 been shown to be useful in the treatment of diseases for the inhibition of
 an overproduction of superoxide and nitric oxide. Thus, the present
 invention relates to the known treatment for diseases by SOD and SOD
 mimics.
 The Beckman et al PCT application also teaches that SODs catalyze the
 dismutation of the oxygen radical superoxide and provides references which
 show SOD and variants thereof have been commonly utilized to prevent or
 reduce oxidation injury in the treatment of stroke and head trauma,
 myocardial ischemia, abdominal vascular occlusion, cystitis, and a variety
 of inflammatory conditions. Beckman et al PCT application also recognizes
 the presence of peroxynitrite in these same disease conditions associated
 with O.sub.2- without indicating the further improvements of the present
 invention.
 Further teachings to the diseases known to be associated with treatment by
 SOD or its mimics are found in EP Publication No. 0524161 (EP Appl. No.
 92870097) which is incorporated by reference therefor.
 Porphyrin complexes are disclosed in U.S. Pat. No. 5,284,674 as valuable
 diagnostic and therapeutic agents, non-peptide phaeophorbide analogs are
 disclosed in Japanese Patent Publication Hei 5-331063 as endocerine
 receptor antagonists, carotenoporphyrins are disclosed in U.S. Pat. No.
 5,286,474 to be valuable for locating and visualizing mammalian tumor
 tissue and similar nitrogen containing macrocycles without a complexed
 metal are disclosed as cytotoxic agents in U.S. Pat. No. 5,283,255. No
 metal complexes and their usefulness are shown as now found in the present
 invention.
 Metal complexes are, however, shown to be useful compounds in Derwent
 Abstract as intermediates in JP05277377-A and MRI agents in U.S. Pat. No.
 5,284,944; cyan pigments in U.S. Pat. No. 5,286,592; photoconductive
 phthalocyanine compositions in U.S. Pat. No. 5,283,146; a recording layer
 in an optical recording medium in U.S. Pat. No. 5,284,943 and near
 infrared absorbers and display/recording materials in an abstract for U.S.
 Pat. No. 5,296,1632.
 Iron hemoprotein is disclosed to be an effective agent to bind or oxidize
 nitric oxide which has a deleterious physiological effect when induced by
 a cytokine or by endotoxin for the treatment of diseases such as septic
 shock in PCT application No. PCT/US93/01288 (Publication No. WO 93/16721).
 Other complexes and their utilities are disclosed. For example, "Ruthenium
 Phthalocyanines" are disclosed as water soluble agents for photodynamic
 cancer Therapy in Platinum Metals Rev., 1995, 39, (1), 14-18; selected
 metallo-organic complexes are disclosed as treatment of inflammation in
 U.S. Pat. No. 4,866,054; Porphyrin and phthalocyanine antiviral
 compositions are disclosed as inhibitors of infection or replication of
 HIV in U.S. Pat. No. 5,109,016; Manganese
 meso-tetra(4-sulfonatophenyl)porphine are synthesized and used as
 tumor-selective MRI contrast agents; an abstract for JP 03273082 teaches
 peroxide-degrading metal porphyrins for use as antioxidants in the
 manufacture of foods or other products; U.S. Pat. No. 4,758,429 teaches
 iron tetraphenyl porphyrin sulfonate acetate for activating magnetic or
 electrical dipoles in the joint with an alternating electromagnetic field
 to treat arthritis and non-infectious joint diseases; an abstract of EP
 392666 shows a non-toxic labile metal atom or complex such as
 1,5,9,13-tetrazacyclohexadecane for use in the treatment of a virus such
 as HIV. CA 119:203240 discloses selected metalloporphyrins as
 hypoplycemics are found in French Patent No. 91-6174. Numerous additional
 references indicate analogous additional uses for metal complexes.
 Finally, nitrogen containing selected macrocycles are shown in JPO5331063
 as endothelin receptor antagonists for treating and preventing
 hypertension, acute renal failure, cardiomyopathy and myocardial
 infarction.
 SUMMARY OF THE INVENTION
 The present invention is a method of treating a disease which is
 advantageously affected by decomposition of peroxynitrite which is
 accelerated over, ie above or more than, a natural background rate of
 decay in humans suffering from the disease comprising administering a
 compound or compound which is a metal complex whereby the peroxynitrite is
 decomposed. Preferably peroxynitrite is decomposed to a benign species.
 The compound is a ligand structure providing a complexed metal, such as
 one of the transition metals, such as Mn, Fe, Ni and V. Preferred ligands
 are macrocyclic ligands, such as porphyrins, aza macrocycles and the like.
 The present invention is a novel method of treating a disease in mammals,
 including humans, advantageously affected by the absence of peroxynitrite
 comprising administration of an accelerated-decomposition effective amount
 of a compound of the formula
 ##STR1##
 wherein
 R.sub.3, R.sub.6, R.sub.9 or R.sub.12 are independently selected a group
 consisting of H, alkyl, alkenyl, CH.sub.2 COOH, phenyl, pyridinyl, and
 N-alkylpyridyl such that phenyl, pyridinyl and N-alkylpyridyl are
 ##STR2##
 which are attached at a carbon atom, and
 wherein phenyl is optionally substituted by halogen, alkyl, aryl, benzyl,
 COOH, CONH.sub.2, SO.sub.3 H, NO.sub.2, NH.sub.2, N(R).sub.3+, wherein R
 is hydrogen, alkyl, or alkylaryl;
 pyridinyl is optionally substituted by halogen, alkyl, aryl, benzyl, COOH
 CONH.sub.2, SO.sub.3 H, NO.sub.2, NH.sub.2, N(R).sub.3+ or NHCOR' wherein
 R is as defined above and R' is alkyl; and
 N-alkylpyridine ring is optionally substituted by halogen, alkyl, aryl,
 benzyl, COOH, CONH.sub.2, SO.sub.3 H, NO.sub.2, NH.sub.2, N(R).sub.3+ or
 NHCOR' wherein R
 and R' are as defined above;
 R.sub.1, R.sub.2, R.sub.4, R.sub.5, R.sub.7, R.sub.8, R.sub.10, or R.sub.11
 are independently selected a group consisting of H, alkyl, alkenyl,
 carboxyalkyl, Cl, Br, F, NO.sub.2, hydroxyalkyl, and SO.sub.3 H or R.sub.1
 and R.sub.2 can be taken together to form a ring of from 5 to 8 carbons
 preferably 6;
 X and Y are suitable ligands or charge-neutralizing anions which are
 derived from any monodentate or polydentate coordinating ligand or ligand
 system or the corresponding anion thereof (for example benzoic acid or
 benzoate anion, phenol or phenoxide anion, alcohol or alkoxide anion) and
 are independently selected from the group consisting of halide, oxo, aquo,
 hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo,
 arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkyl amino,
 heterocycloaryl, amino, amine oxides, hydrazine, alkyl hydrazine, aryl
 hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate,
 isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl
 isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, aryl sulfonic
 acid, alkyl sulfoxide, aryl sulfoxide, alkyl aryl sulfoxide, alkyl
 sulfenic acid, aryl sulfenic acid, alkyl sulfinic acid, aryl sulfinic
 acid, alkyl thiol carboxylic acid, aryl thiol carboxylic acid, alkyl thiol
 thiocarboxylic acid, aryl thiol thiocarboxylic acid, alkyl carboxylic acid
 (such as acetic acid, trifluoroacetic acid, oxalic acid), aryl carboxylic
 acid (such as benzoic acid, phthalic acid), urea, alkyl urea, aryl urea,
 alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl
 thiourea, sulfate, sulfite, bisulfate, bisulfite, thiosulfate,
 thiosulfite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl
 phosphine oxide, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl
 phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide,
 alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl
 phosphinic acid, alkyl phosphinous acid, aryl phosphinous acid, phosphate,
 thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate,
 dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl
 guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl
 thiocarbamate, aryl thiocarbamate, alkyl aryl thiocarbamate, alkyl
 dithiocarbamate, aryl dithiocarbamate, alkyl aryl dithiocarbamate,
 bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite,
 perbromate, bromate, bromite, hypobromite, tetrahalomanganate,
 tetrafluoroborate, hexafluorophosphate, hexafluoroanitmonate,
 hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra
 alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate,
 saccharinate, amino acid, hydroxamic acid, thiotosylate, and anions of ion
 exchange resins, or systems; with the proviso that when the X and Y
 containing complex has a net positive charge then Z is present and is a
 counter ion which is independently X or Y, or when the X and Y containing
 complex has net negative charge then Z is present and is a counter ion
 selected from a group consisting of alkaline and alkaline earth cations,
 organic cations such as alkyl or alkylaryl ammonium cations; and
 M is selected from the group consisting of Mn, Fe, Ni and V;
 ##STR3##
 wherein
 R' is CH or N;
 R.sub.1, R.sub.2, R.sub.3, R4, R.sub.5, R.sub.6, R.sub.7, R.sub.8, R.sub.9,
 R.sub.10, R.sub.11, R.sub.12, R.sub.13, R.sub.14, R.sub.15, and R.sub.16
 are independently selected from a group consisting of H, SO.sub.3 H, COOH,
 NO.sub.2, NH.sub.2, and N-alkylamino;
 X, Y, Z and M are selected as defined above;
 ##STR4##
 wherein
 R.sub.1, R.sub.5, R.sub.9, and R.sub.13 are independently a direct bond or
 CH.sub.2 ;
 R.sub.2, R.sub.2 ', R.sub.4, R.sub.4 ', R.sub.6, R.sub.6 ', R.sub.8,
 R.sub.8 ', R.sub.10, R.sub.10 ', R.sub.12, R.sub.12 ', R.sub.14, R.sub.14
 ', R.sub.16, R.sub.16 ' are independently H, or alkyl;
 R.sub.3, R.sub.7, R.sub.1 l, R.sub.15 are independently H or alkyl;
 X, Y, Z and M are as defined above;
 ##STR5##
 wherein
 R.sub.1, R.sub.5, R.sub.8, and R.sub.12 are independently a direct bond or
 CH.sub.2 ;
 R.sub.2, R.sub.2 ',R.sub.4, R.sub.4 ',R.sub.6, R.sub.6 ', R.sub.7, R.sub.9,
 R.sub.9 ', R.sub.11, R.sub.11 ', R.sub.13, R.sub.13 ', R.sub.14 are
 independently H or alkyl;
 R.sub.3 and R.sub.10 are independently H or alkyl;
 X, Y, Z and M are as defined above;
 ##STR6##
 wherein
 R.sub.1, R.sub.4, R.sub.8, R.sub.12 are independently a direct bond or
 CH.sub.2 ;
 R.sub.2, R.sub.2 ', R.sub.3, R.sub.5, R.sub.5 ', R.sub.7, R.sub.9, R.sub.9
 ', R.sub.11, R.sub.11 ', R.sub.13, R.sub.13 ', R.sub.14 are independently
 H or alkyl;
 R.sub.10 is H or alkyl;
 X, Y, Z and M are as defined above;
 ##STR7##
 wherein
 R.sub.1, R.sub.4, R.sub.7 and R.sub.10 are independently a direct bond or
 CH.sub.2 ;
 R.sub.2, R.sub.2 ', R.sub.3, R.sub.5, R.sub.5 ', R.sub.6, R.sub.8, R.sub.8
 ', R.sub.9, R.sub.11, R.sub.11 ' and R.sub.12 are independently H or
 alkyl;
 X, Y, Z and M are as defined above;
 ##STR8##
 wherein
 R.sub.1, R.sub.4, R.sub.8 and R.sub.11 are independently a direct bond or
 CH2;
 R.sub.2, R.sub.3, R.sub.3 ', R.sub.5, R.sub.5 ', R.sub.7, R.sub.7 ',
 R.sub.9, R.sub.10, R.sub.10 ', R.sub.12, R.sub.12 ' and R.sub.13 are
 dependently H or alkyl;
 R.sub.6 is hydrogen and alkyl;
 X, Y, Z and M are as defined above;
 ##STR9##
 wherein
 R.sub.1, R.sub.4, R.sub.7 and R.sub.10 are independently H or alkyl;
 R.sub.2, R.sub.3, R.sub.3 ', R.sub.5, R.sub.5 ', R.sub.6, R.sub.8, R.sub.9,
 R.sub.9 ', R.sub.11, R.sub.11 ' and R.sub.12 are independently H or alkyl;
 X, Y, Z and M are as defined above;
 ##STR10##
 wherein
 R.sub.1, R.sub.3, R.sub.4 and R.sub.6 are independently H or alkyl;
 R.sub.2 and R.sub.5 are independently selected from the group consisting of
 H, alkyl, SO.sub.3 H, NO.sub.2, NH.sub.2, halogen, COOH, and N(R).sub.3+
 wherein R is as defined above;
 X, Y, Z and M are as defined above;
 ##STR11##
 wherein
 R.sub.1, R.sub.2, R.sub.3, R.sub.4 are independently selected from the
 group consisting of H, alkyl, SO.sub.3 H, NO.sub.2, NH.sub.2, halogen,
 COOH and N(R).sub.3+ wherein R is as defined above;
 X, Y, Z and M are as defined above;
 ##STR12##
 wherein
 R1, R1', R2, R2', R3, R3', R4, R4', R5, R5', R6, R6', R7 and R7' are
 independently selected from a group consisting of H, alkyl, alkoxy,
 NO.sub.2, aryl, halogen, NH.sub.2, SO.sub.3 H, and R.sub.6, R.sub.6 ',
 R.sub.7 and R.sub.7 ' may each be taken together with one other of
 R.sub.6, R.sub.6 ', R.sub.7 and R.sub.7 ' to form a cyclic group,
 preferably a 6 carbon cycloalkyl group;
 M.sup.1 is Fe, Ni or V;
 X, Y and Z are as defined above together with a pharmaceutically acceptable
 carrier, preferably in unit dosage form.
 The present invention is also a pharmaceutical composition for the
 treatment of a disease in humans advantageously affected by accelerated
 decomposition over the natural background rate of decay of peroxynitrite
 comprising an amount effective for the accelerated decomposition of
 peroxynitrite in humans of a compound of the formula I, II, IIIA, IIIB,
 IIIC, IIID, IIIE, IIIF, IIIG, IIIH as defined above with a
 pharmaceutically acceptable carrier in unit dosage form, preferably oral
 unit dosage form.

DETAILED DESCRIPTION OF THE INVENTION
 As utilized herein, the term "alkyl", alone or in combination, means a
 straight-chain or branched-chain alkyl radical containing from 1 to about
 22 carbon atoms, preferably from about 1 to about 18 carbon atoms, and
 most preferably from about 1 to about 12 carbon atoms. Examples of such
 radicals include, but are not limited to methyl, ethyl, n-propyl,
 isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, iso-amyl,
 hexyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl and
 eicosyl. The term "aryl", alone or in combination, means a phenyl or
 naphthyl radical which optionally carries one or more substituents
 selected from alkyl, cycloalkyl, cycloalkenyl, aryl, heterocycle,
 alkoxyaryl, alkaryl, alkoxy, halogen, hydroxy, amine, cyano, nitro,
 alkylthio, phenoxy, ether, trifluoromethyl and the like, such as phenyl,
 p-tolyl, 4-methoxy-phenyl, 4-(tert-butoxy)phenyl, 4-fluorophenyl,
 4-chlorophenyl, 4-hydroxyphenyl, 1-naphthyl, 2-naphthyl, and the like. The
 term "aralkyl", alone or in combination, means an alkyl or cycloalkyl
 radical as defined herein in which one hydrogen atom is replaced by an
 aryl radical as defined herein, such as benzyl, 2-phenylethyl, and the
 like. The term "heterocyclic" means ring structures containing at least
 one other kind of atom, in addition to carbon, in the ring. The most
 common of the other kinds of atoms include nitrogen, oxygen and sulfur.
 Examples of heterocyclics include, but are not limited to, pyrrolidinyl,
 piperidyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl,
 thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl,
 imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl,
 benzothiadiazolyl, triazolyl and tetrazolyl groups. The term "cycloalkyl",
 alone or in combination means a cycloalkyl radical containing from 3 to
 about 10, preferably from 3 to about 8, and most preferably from 3 to
 about 6 carbon atoms. Examples of such cycloalkyl radicals include, but
 are not limited to, cyclopropyl, cyclobutyl, cyclophetyl, cyclohexyl,
 cycloheptyl, cyclooctyl, and perhydronaphthyl. The term "cycloalkenyl",
 alone or in combination, means a cycloalkyl radical having one or more
 double bonds. Examples of cycloalkenyl radicals include, but are not
 limited to cyclopentenyl, cyclohexenyl, cycloooctenyl, cyclopentadienyl,
 cyclohexadienyl, and cyclooctadienyl.
 The macrocyclic ligands useful in the present invention wherein the formula
 is Structure I can be prepared according to the general synthetic methods
 known in the art for preparation of certain ligands. See, for example,
 1) Campestrini, S.; Meunier, B. Inorg. Chem. 31, 1999-2006, (1992).
 2)Robert, A.; Loock, B.; Momenteau, M.; Meunier, B. Inorg. Chem. 30,
 706-711, (1991).
 3)Lindsey, J. S.; Wagner, R. W. J. Org. Chem. 54, 828-836, (1989).
 4) Zipplies, M. F.; Lee, W. A.; Bruice, T. C. J. Am. Chem. Soc. 108,
 4433-4445, (1986).
 The macrocyclic ligands useful in the present invention wherein the formula
 is Structure II can be prepared according to the general synthetic methods
 known in the art for preparation of certain ligands. See, for example,
 1) Some compounds are commercially available from Porphyrin Products, Inc.
 (Logan, Utah.)
 2) Y. L. Meltze; Phthalocyanine Technology in Chemical Process Reviews No.
 42.; Noyes Data Corp, Park Ridge, N.J. (1970).
 The macrocyclic ligands useful in the present invention wherein the formula
 is Structure III can be prepared according to the general synthetic
 methods known in the art for preparation of certain ligands. See, for
 example,
 1)Goedken, V. L.; Molin-Case, J.; Whang, Y-A; J.C.S.Chem.Comm. 337-338,
 (1973)
 2) Martin, J. G.; Cummings, S. C.; Inorg.Chem. 12, 1477-1482, (1973).
 3)Riley, D. P.; Stone, J. A.; Busch, D. H. J.Am.Chem.Soc. 98, 1752-1762,
 (1976).
 4)Dabrowiak, J. C.; Merrell, P. H.; Stone, J. A.; Busch, D. H.;
 J.Am.Chem.Soc. 95, 6613-6622, (1973).
 5) Riley, D. P.; Busch, D. H.; Inorg. Chem. 23, 3235-3241, (1984).
 6)Watkins, D. D.; Riley, D. P.; Stone, J. A.; Busch, D. H.; Inorg. Chem.
 15, 387-393, (1976).
 7)Riley, D. P.; Stone, J. A.; Busch, D. H.; J.Am.Chem.Soc. 99, 767-777,
 (1977).
 The macrocyclic ligands useful in the present invention wherein the formula
 is Structure IV can be prepared according to the general synthetic methods
 known in the art for preparation of certain ligands. See, for example,
 1) Diehl, H.; Hoch, C. C.; Inorganic Synthesis Vol 3. p 196. McGraw-Hill,
 New York (1950).
 2) Srinivasan, K; Michaud, P.; Kochi, J. K; J. Am. Chem.Soc. 108,
 2309-2320, (1986).
 3) Samsel, E. G.; Srinivasan, K.; Kochi, J. K J. Am. Chem. Soc. 107,
 7606-7617, (1985).
 The compounds of the present invention can possess one or more asymmetric
 carbon atoms and are thus capable of existing in the form of optical
 isomers as well as in the form of racemic or nonracemic mixtures thereof.
 The optical isomers can be obtained by resolution of the racemic mixtures
 according to conventional processes, for example by formation of
 diastereoisomeric salts by treatment with an optically active acid.
 Examples of appropriate acids are tartaric, diacetyltartaric,
 dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid and then
 separation of the mixture of diastereoisomers by crystallization followed
 by liberation of the optically active bases from these salts. A different
 process for separation of optical isomers involves the use of a chiral
 chromatography column optimally chosen to maximize the separation of the
 enantiomers. Still another available method involves synthesis of covalent
 diastereoisomeric molecules by reacting one or more secondary amine
 group(s) of the compounds of the invention with an optically pure acid in
 an activated form or an optically pure isocyanate. The synthesized
 diastereoisomers can be separated by conventional means such as
 chromatography, distillation, crystallization or sublimation, and then
 hydrolyzed to deliver the enantiomerically pure ligand. The optically
 active compounds of the invention can likewise be obtained by utilizing
 optically active starting materials, such as natural amino acids.
 To screen metal complexes for peroxynitrite decomposition catalytic
 activity of the present invention, peroxynitrite is prepared and isolated
 as its sodium salt by the reaction of acidic hydrogen peroxide with sodium
 nitrite followed by rapid quenching with NaOH as set out by Halfpenny and
 Robinson, in J. Chem. So., 1952, 928-938. Peroxynitrite has an absorbance
 maximum at 302 nm with an extinction coefficient of 1670 M.sup.-1
 cm.sup.-1. Therefore, it is possible to directly observe the decomposition
 of peroxynitrite by stop-flow spectrophotometric analysis by monitoring
 the decomposition of the absorbance at 302 nm. That is, such observation
 of the decomposition of peroxynitrite at a rate accelerated over the
 natural decomposition rate with the addition of the metal complex
 identifies a compound of the present invention.
 In addition, it is now found that peroxynitrite inactivates CuZnSOD enzyme
 in a concentration dependant manner. Since it is known peroxynitrite also
 inactivates MnSOD (See "Peroxynitrite-Mediated Tyrosine Nitration
 Catalyzed by Superoxide Dismutase" by Ischiropoulos et al in Archives of
 Biochemistry and Biophysics, Vol. 298, No. 2, November 1, pp. 431-437,
 1992), the present invention provides a compound which protects CuZnSOD
 from inactivation by peroxynitrite.
 In this manner the compound of the present invention is shown to be useful
 in treating a disease in a human advantageously affected by the presence
 of the SOD enzyme.
 That is, the treatment of the present invention is for a disease state
 either caused by the presence of a peroxynitrite of caused by the lack of
 the protective presence of the SOD enzyme such as in a myocardial infarct,
 stroke or an autoimmune disease. These latter diseases are also shown to
 be associated with the presence of peroxynitrite.
 These metal complexes are found to be within the present invention by
 determination of their decomposition effect on peroxynitrite as set out
 herein.
 Contemplated equivalents of the general formulas set forth above for the
 compounds and derivatives as well as the intermediates are compounds
 otherwise corresponding thereto and having the same general properties
 such as tautomers of the compounds and such as wherein one or more of the
 various R groups are simple variations of the substituents as defined
 therein, e.g., wherein substituents which are a higher alkyl group than
 that indicated, or where the tosyl groups are other nitrogen or oxygen
 protecting groups or wherein the O-tosyl is a halide. Anions having a
 charge other than 1, e.g., carbonate, phosphate, and hydrogen phosphate,
 can be used instead of anions having a charge of 1, so long as they do not
 adversely affect the overall activity of the complex. However, using
 anions having a charge other than 1 will result in a slight modification
 of the general formula for the complex set forth above. In addition, where
 a substituent is designated as, or can be, a hydrogen, the exact chemical
 nature of a substituent which is other than hydrogen at that position,
 e.g., a hydrocarbyl radical or a halogen, hydroxy, amino and the like
 functional group, is not critical so long as it does not adversely affect
 the overall activity and/or synthesis procedure.
 The chemical reactions shown by the references described above are
 generally disclosed in terms of variations appropriate for their broadest
 application to the preparation of the compounds of this invention.
 Occasionally, the reactions may not be applicable as described to each
 compound included within the disclosed scope. The compounds for which this
 occurs will be readily recognized by those skilled in the art. In all such
 cases, either the reactions can be successfully performed by conventional
 modifications known to those skilled in the art, e.g., by appropriate
 protection of interfering groups, by changing to alternative conventional
 reagents, by routine modification of reaction conditions, and the like, or
 other reactions disclosed herein or otherwise conventional, will be
 applicable to the preparation of the corresponding compounds of this
 invention. In all preparative methods, all starting materials are known or
 readily preparable from known starting materials.
 Without further elaboration, it is believed that one skilled in the art
 can, using the preceding description, utilize the present invention to its
 fullest extent. The following preferred specific embodiments are,
 therefore, to be construed as merely illustrative, and not limitative of
 the remainder of the disclosure in any way whatsoever.
 EXAMPLES
 All reagents were used as received unless otherwise indicated.
 5,10,15,20-tetrakis(N-Methyl-4-pyridyl)porphyrin tetratosylate and
 Acetato-5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrin iron(III) were
 purchased from Porphyrin Products Inc. (Logan, Utah). Iron(III)citrate and
 iron(III)EDTA complexes were purchased from Aldrich Chemical Co.
 (Milwaukee, Wis.). All nuclear magnetic resonance (NMR) spectra were
 obtained on Varian VXR-300 or Varian VXR-400 spectrometers. Qualitative
 and quantitative mass spectra were run on a Finnigan MAT90, a Finnigan
 4500 and a VG40-250T spectrometers.
 Example 1
 Synthesis of Acetato (5,10,15,20-tetrakis(N-methyl-4-pyridyl)porphinato)
 iron (III) tetra-tosylate, Fe(III)TMPyP.
 5,10,15,20-Tetra-(N-methyl-pyridyl)porphine tetratosylate, (H.sub.2 TMPyP)
 (0.30 g, 0.231 mmole) was charged to a 100 mL round bottom flask equipped
 with a magnetic stir bar and was dissolved in a minimal amount of MeOH.
 Anhydrous Fe(OAc).sub.2 (0.120 g, 0.692 mmole) was added followed
 immediately by 25 mL of glacial acetic acid and 100 uL of triethylamine.
 The reaction mixture was heated to reflux. The reaction was monitored by
 visible spectroscopy and was determined to be complete with the appearance
 of a strong band at 426 nm indicative of the metallated porphyrin. The
 MeOH was removed by evaporation and the solid was taken up again in a
 minimal amount of MeOH. The mixture was concentrated under vacuum to a
 total volume of .about.20 mL at which point the unreacted Fe(OAc).sub.2
 precipitates. The solid was separated by centrifugation and the mother
 liquor is chromatographed on a Sephadex LH-20 column (2.times.30 cm) using
 MeOH as eluent. The initial colored band was collected and
 Fe(III)TMPyP(OAc) was isolated by precipitation after evaporation of
 solvent and trituration with ether to give 85 mg (26%) of the desired
 product as confirmed by mass spectral analysis.
 Example 2
 Synthesis of 5,10,15,20-tetrakis(3,5-disulfonatedmesityl)porphyrin
 octasodium salt (H.sub.2 TMPS).
 5,10,15,20-tetramesitylporphyrin (H.sub.2 TMP) was prepared by the
 condensation of pyrrole and mesitaldehyde in sealed glass tubes by the
 method of Badger (G. M. Badger, R. A. Jones, R. L. Laslett Aust. J. Chem.,
 17, 1022, [1964]) or in refluxing collidine according to the literature
 preparation of Meunier (Meunier et. al. Nouv. J. Chim., 10, 39-49,
 [1986]). Chlorin impurities were removed by oxidation with
 2,3-dichloro-5,6-dicyano-1,4,-benzoquinone in refluxing benzene followed
 by chromatography on basic alumina. Both methods produced nearly identical
 yields of H.sub.2 TMPS.
 Synthesis of H.sub.2 TMPS was achieved using a slight modification of the
 method of Meunier (Meunier et. al. Inorg. Chem, 31, 1999-2006, [1992]). A
 25 mL round bottom flask equipped with a reflux condenser and stir bar was
 charged with H.sub.2 TMP (1.0 g, 1.28 mmole). Oleum (H.sub.2 SO.sub.4
 +18-23% SO.sub.3) 10 mL was added and the reaction was heated to
 80.degree. C. for 40 min. The reaction was cooled and its contents was
 added dropwise to 100 mL of water cooled in an ice bath. The resulting
 water solution was neutralized with 2N NaOH (.about.220 mL) to a pH=6-7.
 The water was removed by evaporation and the resulting solid residue was
 triturated with a minimal amount of MeOH. The resulting precipitate was
 removed by filtration and the filtrate was further concentrated to 60 mL
 under vacuum. The resulting precipitate (additional Na.sub.2 SO.sub.4) was
 separated by centrifugation. The supernatant was evaporated to dryness
 generating 1.59 g (78%) of the desired sulfonated porphyrin.
 Example 3
 Synthesis of Acetato 5,10,15,20tetrakis(3,5,disulfonatomesityl)porphyrin
 Manganese(III) octasodium salt (Mn(III)TMPS).
 H.sub.2 TMPS (0.2 g, 0.125 mmole) and Mn(OAc).sub.2 (0.296 g, 1.71 mmole)
 was dissolved in 38 mL of water and was heated to 85.degree. C. for 1 h.
 The reaction was monitored by visible spectroscopy and was determined to
 be complete when the Soret band (416 nm) of the free base porphyrin was
 replaced by a new band at 468 nm characteristic of Mn(III) porphyrin
 species. The reaction was reduced in volume under vacuum to 10 mL and was
 chromatographed on a Dowex 50WX-8 cation exchange resin (H.sup.+ form) to
 remove excess Mn(OAc).sub.2. The eluent was reduced in volume to 10 mL and
 was adjusted to pH=8.0 with 1.0 N NaOH. The resulting solution was
 evaporated to dryness. The residue was taken up in 7 mL of MeOH and
 chromatographed on a Sephadex LH-20 column using MeOH as eluent. The
 purple band was collected and evaporated to dryness giving 0.175 g(90%) of
 the desired metallated porphyrin as determined by mass spectral analysis.
 Example 4
 Synthesis of Acetato-5,10,15,20-tetrakis(3,5-disulfonatomesityl)porphyrin
 Iron (III) octasodium salt (Fe(III)TMPS).
 H.sub.2 TMPS (0.2 g, 0.125 mmole) and Fe(OAc).sub.2 (0.300 g 1.72 mmole)
 was dissolved in 38 mL of water. The reaction mixture was brought to
 reflux and was monitored by visible spectroscopy to determine complete
 metallation. Upon completion the reaction was filtered and reduced in
 volume to 10 mL. The orange-brown reaction mixture was passed through a
 Dowex 50WX-8 cation exchange column (H+from) to remove excess
 Fe(OAc).sub.2. The eluent was reduced in volume to 10 mL and was adjusted
 to a pH=7.5 with 1.0 N NaOH. The resulting solution was evaporated to
 dryness. The residue was taken up in 7 mL of MeOH and chromatographed on a
 Sephadex LH-20 column using MeOH as eluent. The orange-brown band was
 evaporated to dryness giving 0.170 g (72%) of the desired Fe porphyin as
 confirmed by mass spectral analysis.
 Example 5
 Synthesis of Acetato-5,10,15,20-tetrakis(3,5-disulfonatomesityl)porphyrin
 Nickel (II) octasodium salt (Ni(II)TMPS).
 H.sub.2 TMPS (0.1 g, 0.063 mmole) and Ni(OAc).sub.2 (0.156 g, 0.63 mmole)
 was dissolved in 20 mL of water and was refluxed for 3 h. The reaction
 mixture was orange in color indicative of the Ni porphyrin. The completion
 of the reaction was confirmed by Vis spectroscopy. The reaction was
 reduced in volume to 5 mL and chromatographed on a Dowex 50 WX-8 ion
 exchange column (H.sup.+ form) to remove excess Ni(OAc).sub.2. The eluent
 was reduced in volume to 5 mL and was adjusted to a pH=8.0 with 1.0 N
 NaOH. The resulting solution was evaporated to dryness. The residue was
 taken up in 7 mL of MeOH and chromatographed on a Sephadex LH-20 column
 using MeOH as eluent. Product was isolated by removal of solvent to give
 0.090 g (85%) of the desired metallated porphyrin as confirmed by mass
 spectral analysis.
 Example 6
 Synthesis of N,N'-ethylenebis(3,3'dimethoxysalicylideneamine) ligand.
 A modification of the procedure of Coleman was used (Coleman et al. Inorg.
 Chem , 20, 700, [1981]). A 100 mL round bottom flask equipped with a stir
 bar was charged with 25 mL of absolute EtOH and 3-methoxysalicyladehyde
 (3.04 g, 0.02 mol). A 20 mL solution of absolute EtOH and ethylenediamine
 (0.601 g, 0.01 mol) was freshly prepared and was added in one portion to
 the salicylaldehyde. The reaction was refluxed for 1 h during which time a
 yellow-orange precipitate appeared. The product was collected by
 filtration, washed with 100 mL of hot ethanol, and dried under vacuum to
 give 4.4 g (98%) of the desired product.
 Example 7
 Synthesis of Chloro[N,N'-ethylenebis(3,3'-dimethoxysalicylideneaminato)iron
 (III)
 N,N'-Ethylenebis(3,3'dimethoxysalicylideneamine) (0.05 g, 0.188 mmole) was
 dissolved in 20 mL of MeOH and Fe(Cl).sub.3 (0.030 g, 0.188 mmole) was
 added in one portion. The solution was refluxed for 1 h after which time
 the solvent was removed under vacuum. The purple residue was washed with a
 minimal amount of water. The solid was taken up in 10 mL of MeOH, filtered
 and re-isolated by removal of solvent to give 0.047 g (70%) of the desired
 iron complex.
 Example 8
 Synthesis of
 12,14-Dimethyl-1,4,8,11-tetraazacyclotetradeca-11,13-dienatonickel(II)
 Hexaflorophosphate, Ni(II)([14]dienoN.sub.4)PF.sub.6
 Ni(II)([14]dienoN.sub.4)PF.sub.6 was prepared by the method of Martin and
 Cummings (Martin, J. G.; Cummings, S. C. Inorg. Chem., 12, 1477-1482,
 [1973]). The compound was characterized by mass spectral analysis and was
 shown to be consistent with the desired structure.
 Example 9
 Synthesis of
 12,14-Dimethyl-1,4,8,11-tetraazacyclotetradeca-11,14-dienenickel(II)
 Hexaflorophosphate, Ni(II)([14]dieneN.sub.4)(PF.sub.6).sub.2
 Ni(II)([14]dieneN.sub.4)(PF.sub.6).sub.2
 Ni(II)([14]dieneN.sub.4)(PF.sub.6).sub.2 was prepared from
 Ni(II)([14]dienoN.sub.4)PF.sub.6 by the method of Martin and Cummings (
 Martin, J. G.; Cummings, S. C. Inorg. Chem., 12, 1477-1482, [1973]).
 Example 10
 Synthesis of 6,8,15,17-Tetramethyldibenzo[b,i][1,4,8,11
 tetraazatetradeca-2,4,7,9,12,14-hexaenatonickel(II), Ni(II)[14]12eneN.sub.4
 Ni(II)[14]12eneN.sub.4 was prepared by the method of Goendken et. al.
 (Goendken et. al. J.C.S. Chem.Comm., 337-338, [1973]). The complex was
 characterized by mass spectral analysis and which was consistent with the
 desired structure.
 Example 11
 This example describes the preparation of peroxynitrite stock solutions
 used in these studies. A modified version of the procedure described by
 Hughs was used (Hughs, M. N.; Nicklin, H. G. J. Chem. Soc., (A), 450-452,
 [1968]).
 To 10 mL of vigorously stirred 0.6 M NaNO.sub.2 solution maintained at
 0.degree. C. was added an equal volume of a HCl/H.sub.2 O.sub.2 solution
 (0.6 M HCl and 0.7 M H.sub.2 O.sub.2) followed immediately by the rapid
 addition of 10 mL of 0.75 M NaOH. The resulting yellow solution was
 treated with 25 mg of MnO.sub.2 for 3 min. and was immediately filtered.
 The filtrate was placed in a -20.degree. C. freezer for several days which
 resulted in the fractionation of the sodium peroxynitrite as evident by a
 fine yellow band visible at the top of the flask. The yellow band was
 collected to yield .about.1 mL of a 280 mM sodium peroxynitrite solution.
 This solution could be stirred frozen at -20.degree. C. for several days
 with minimal decomposition of peroxynitrite.
 Example 12
 This example describes the procedures used to determine if compounds are
 peroxynitrite decomposition catalyst by stopped-flow kinetic analysis.
 All analysis were run using potassium phosphate buffers (Calbiochem) which
 were biological grade using ultra pure water prepared by the method of
 Riley (Riley, D. P. et. al. Anal. Biochem. 196, 344-349, [1991]). Kinetic
 measurements were made on an OLIS Rapid Scanning Stopped-Flow Spectrometer
 (On-Line Instrument Systems Inc., Bogart, Ga.)) using the OLISRSM-1000
 Operating system for data acquisition and manipulation. Peroxynitrite has
 a strong absorbance at 302 nm (extinction coefficient =1670 M-1 cm-1) and
 has been shown to decompose in a process that is first-order in sodium
 peroxynitrite and first order in protons (Hughs, M. N.; Nicklin, H. G. J.
 Chem. Soc., (A), 450-452, [1968]) with t.sub.1/2 =1.9 sec. at 37.degree. C
 pH=7.4 (Beckman, J. S. et.al. Proc. Natl. Acad. Sci. USA, 87, 1620-1624,
 [1990]).
 Thus, in a typical experiment the natural background decomposition rate of
 sodium peroxynitrite was determined as follows. A 24 mM stock solution of
 sodium peroxynitrite in 50 mM NaOH is load into the small volume syringe
 and 100 mM potassium phosphate pH=7.4 is charged into the large volume
 syringe of the stopped-flow spectrophotometer. All stopped -flow
 measurements were made at 22 .degree. C. Injection of the solutions into
 the sample compartment results in .about.25 fold dilution of the stock
 sodium peroxynitrite. The decomposition of sodium peroxynitrite is first
 order in peroxynitrite with a t.sub.1/2 =5.2 see and a k.sub.obs
 =1.39.times.10.sup.-1.+-.0.15 sec.sup.-1. To test for catalytic
 peroxynitrite decomposition activity, the metal complex was dissolved in
 100 mM potassium phosphate buffer pH=7.4 and loaded into the large syringe
 and the decomposition of peroxynitrite was monitored as described above.
 The catalytic rate constant (k.sub.cat M-1 sec.sup.-1) for the complexes
 tested was determined by varying the complex concentration and plotting
 k.sub.obs vs [complex] Table 1. The k.sub.obs were obtained from averages
 of three stopped flow analysis at each catalyst concentration. Data
 representative of this analysis for a variety of compounds is shown in
 FIG. 1. The simple di and trivalent chloride salts of Mn, Fe, Co, Cu, and
 Ni showed no catalytic peroxynitrite decomposition activity at
 concentration of 0.050 mM and below.
 TABLE 1
 CATALYTIC RATE CONSTANTS FOR THE
 DECOMPOSITION OF SODIUM PEROXYNITRITE BY METAL
 COMPLEXES AT pH = 7.4 AND 22.degree. C.
 Example No. Complex k.sub.cat (M.sup.-1
 sec.sup.-1)
 1 Fe(III)TMPyP 2.75 .times. 10.sup.+6
 Fe(III)TPPS 2.06 .times. 10.sup.+6
 4 Fe(III)TMPS 1.60 .times. 10.sup.+5
 5 Ni(II)TMPS 8.72 .times. 10.sup.+4
 7 Fe(III)(3,3'MeO.sub.2 Salen) 5.00 .times. 10.sup.+4
 3 Mn(III)TMPS 2.90 .times. 10.sup.+4
 8 Ni(II)([14]dienoN.sub.4)PF.sub.6 2.05 .times.
 10.sup.+4
 9 Ni(II)([14]dieneN.sub.4)(PF.sub.6).sub.2 1.80 .times.
 10.sup.+4
 10 Ni(II)[14]12eneN.sub.4 1.70 .times. 10.sup.+4
 Fe(III)EDTA 2.00 .times. 10.sup.+4
 Fe(III)Citrate 1.50 .times. 10.sup.+4
 2 H2TMPS Inactive
 1(SM).sup.b H2TMPyP Inactive
 ZnTMPyP Inactive
 Ni(CR)Cl.sub.2.sup.a Inactive
 .sup.a CR =
 2,12-dimethyl-3,7,11,17-tetraazabicyclo[11.3.
 1]heptadeca-1(17),2,11,13,15-pentaene
 .sup.b Starting material
 Example 13
 This example illustrates the inactivation of CuZn-superoxide dismutase
 (CuZnSOD) by peroxynitrite and that peroxynitrite decomposition catalyst
 shown to be active in Example 12 protect CuZnSOD against inactivation by
 peroxynitrite.
 Stock solutions of bovine liver CuZnSOD (DDI Pharmaceuticals Inc., Mountain
 View CA) were prepared by dissolving -1.0 mg of enzyme in 10 mL of 50 mM
 potassium phosphate buffer at a pH=7.4. The activity of this solution to
 dismutate superoxide was determined by the method of Riley (Riley, D. P.
 et. al. Anal. Biochem. 196, 344-349, [1991]). All k.sub.obs were the
 average of triplicate runs using a stopped flow spectrophotometer
 manufactured by Kinetic Instruments Inc. (Ann Arbor, Mich.) and was
 interfaced to a MAC IICX personal computer.
 Inactivation of CuZn SOD by peroxynitrite.
 Inactivation of peroxynitrite was performed by aloquating 1.0 mL of stock
 CuZn SOD solution into 50 mM potassium phosphate buffer pH=7.4 such that a
 final assay volume of 10 mL is achieved after addition of peroxynitrite
 and EDTA solutions. To these assay solution was added various amounts of
 peroxynitrite (25 mM stock solution) such that the final concentration of
 peroxynitrite in the assay varied from 0, 25, 50, 75 and 100 uM. After the
 addition of peroxynitrite, 100 uL of a 2.5 mM stock EDTA solution was
 added to each assay solution such that the final concentration of EDTA was
 250 uM. Each solution was then assayed by stopped flow analysis for
 superoxide dismutase activity. A plot of k.sub.obs vs peroxynitrite
 concentration is shown in FIG. 2. Control reaction which contained CuZnSOD
 in the presence of 250 uM EDTA alone and 100 uM potassium nitrite or
 nitrate showed no decrease in CuZnSOD activity.
 Example 14
 Protection of CuZnSOD from inactivation by peroxynitrite using
 peroxynitrite decomposition catalysts
 Assay solutions were prepared as described above except for the addition of
 various of peroxynitrite decomposition catalyst. The final solution volume
 was maintained at 10 mL. Thus, to the assay solutions Fe(III)TMPyP (0.5
 and 1.0 uM final concentration) and Fe(III)TMPS (1.0 and 5.0 uM final
 concentration) was added. The solution were then treated with various
 amounts of peroxynitrite such that the final concentrations of 0,25, 50,
 75 and 100 uM were achieved. Following treatment with peroxynitrite EDTA
 was added to a final concentration of 250 uM. The solutions were then
 assayed for SOD activity. Plots of k.sub.obs vs [peroxynitrite] at various
 catalysts concentrations illustrates the protective effect of Fe(III)TMPyP
 FIG. 3 and Fe(III)TMPS FIG. 4. Under the assay conditions employed,
 Fe(III)TMPyP and Fe(II)TMPS were shown not to be effective catalysts for
 the dismutation of superoxide.
 Example 15
 In Vitro Evaluation:
 Serials: Human recombinant tumor necrosis factor-alpha (TNF-a) was obtained
 from Genzyme Corporation, Cambridge, MA. Human recombinant complement C5a
 and L-arginine (L-arg) was purchased from Sigma Chemical Company, St.
 Louis, Mo. Authentic peroxynitrite in 50 mM NaOH was prepared as described
 above.
 Isolation of Endothelial Cells: Human dermal microvascular endothelial
 cells (HDME cells) from neonatal foreskin were prepared as previously
 described (Marks, R. M., Czerniecki, M., and Penny, R. In Vit. Cell.
 Devel. Biol., 21, 627-635 [1985]). In brief, neonatal foreskin tissue from
 several donors was washed in 70% ethanol, cut into small pieces, then
 emersed in trypsin (0.6%; Irvine Scientific, Santa Ana, Calif.) and EDTA
 (1%; Sigma Chemical Company, St. Louis, Mo.) for 7-9 minutes. The
 endothelial cells were removed by pressing the unkeratinized surface of
 the tissue with a scalpel blade. The cells were centrifuged through a 35%
 Percoll density gradient (Sigma Chemical Company, St. Louis, Mo.). After
 centrifugation at 250.times.g for 10 min, cells corresponding to a density
 of less than 1.048 g/ml were collected and plated onto gelatin coated
 tissue culture dishes (0.1%; Sigma Chemical Company, St. Louis, Mo.).
 Contaminating cells were weeded daily using a 25 gauge needle mounted onto
 a tuberculin syringe. Purified endothelial cells were grown to passage 5
 (.about.8 population doublings) in MCDB 131 (Endothelial basal medium;
 Clonetics Corporation) supplemented with 30% human serum (BioWittaker,
 Inc., Walkersville, Md.), 10 ng/ml EGF (Collaborative Biomedical Products,
 Bedford, Mass.), 2 mM L-glutamine (Irvine Scientific, Santa Ana, Calif.),
 and 250 .mu.g/ml dibutyryl CAMP, 1 .mu.g/ml hydrocortisone (Sigma Chemical
 Company, St. Louis, Mo.). These cells were characterized as normal
 endothelial cells by testing for endothelial cell markers (Factor VIII
 immunoreactivity, cell-associated angiotensin converting enzyme activity,
 and low density lipoprotein uptake). Cells were cryopreserved at passage 5
 in 10% DMSO for use in the subsequent assays after testing negative for
 mycoplasma (Coriel Institute, Camden, N.J.).
 Preparation of Neutrophils: Human neutrophils were isolated from peripheral
 blood of healthy donors (Look, D. C., Rapp, S. R., Keller, B. T., and
 Holtzman, M. J. Am. J. Physiol., 263, L79-L87 [1992]). EDTA
 anti-coagulated blood was separated using a single-step density
 centrifugation (PMN Prep, Robbins Scientific, Sunnyvale, Calif.) followed
 by several washes in Hank's buffered saline solution (HBSS; Sigma Chemical
 Company, St. Louis, Mo.) and hypotonic lysis of erythrocytes. Preparations
 contained &gt;95% neutrophils and were &gt;95% viable by trypan blue (GIBCO
 Laboratories, Grand Island N.Y.) exclusion. Purified neutrophils were
 suspended in HBSS supplemented with 0.01% BSA (Miles, Inc., Kankakee,
 Ill.) and 300 uM L-arg (HBSSBA) at a concentration of 5.times.10.sup.6
 cells/ml.
 Endothelial Cell Injury Assays: The cytotoxic effects of stimulated
 neutrophils or peroxynitrite on endothelial cells was determined using a
 .sup.51 Cr-release assay as described by Moldow (Moldow et. al. Methods
 Enzymol., 105, 378-385, [1984]). Passage 5 HDME cells were grown to a
 density of .about.1-2.times.10.sup.4 cells/cm.sup.2 (.about.90%
 confluence) in 96 well microtiter plates and labeled for 18 h with 10
 uCi/ml sodium [.sup.51 Cr]chromate (Amersham Corporation, Arlington
 Heights, Ill.). The HDME cells were cytokine-activated for 4 h with 100
 U/ml human recombinant tumor necrosis factor-alpha (TNF-a; Genzyme
 Corporation, Cambridge, Mass.), then washed twice with HBSSBA. Suspensions
 of neutrophils were added at a concentration of 2.5.times.10.sup.5 /well
 and allowed to settle for 15 min. Unless otherwise noted, the neutrophils
 were activated by priming with 25 U/ml TNF-a for 10 min followed by
 activation with 3 .mu.g/ml complement component C5a (Sigma Chemical
 Company, St. Louis, Mo.). Incubations were continued for 2 h at 37.degree.
 C. When authentic peroxynitrite was used, it was added in the absence of
 neutrophils. Peroxynitrite was added directly to the HDME cell layer from
 a 25 mM stock in 50 mM NaOH giving a final concentration from 0-800 uM.
 All inhibitors were made fresh immediately prior to the assay in HBSSBA
 and added as 1/10 of the well volume before peroxynitrite addition or
 neutrophil activation.
 .sup.51 Cr release was determined by aspiration of the supernatant from
 each well (soluble fraction). The monolayers were washed gently with
 HBSSBA to remove non-adherent cells and the washes pooled with the soluble
 fraction. The adherent cells from each well were solubilized with 1 N NaOH
 and removed to a separate tube. Both fractions were analyzed by gamma
 scintillation spectrometry. Results were expressed as percent .sup.51 Cr
 release as follows: % release=cpm (soluble+nonadherent/total cpm per
 well).times.100. Specific cytotoxicity reflects the difference between
 .sup.51 Cr release induced by stimulated neutrophils and unstimulated
 neutrophils (typically 1-2% above spontaneous release). Results were
 confirmed in 2-3 separate assays and the data presented are
 representative.
 As can be seen from FIG. 5, addition of peroxynitrite to endothelial cells
 results in a dose dependent increase in cell injury demonstrating the
 cytotoxic effects of peroxynitrite. Complexes which have been shown to be
 peroxynitrite decomposition catalysts by stopped flow analysis are capable
 of protecting against peroxynitrite mediated cell injury FIG. 6. These
 complexes are also capable of protecting against neutrophil mediated cell
 injury in a dose dependant fashion FIG. 7.
 Example 16
 Protocol for Cell Protection Assays using Peroxynitrite Decomposition
 Catalysts: A cell viability assay was established to rapidly assess the
 efficacy of peroxynitrite(PN) catalysts in protecting cells from PN25
 mediated injury and death. The peroxynitrite challenge consisted of a
 pulse of synthetic PN added exogenously to cells. In order to better
 assess the efficacy of PN catalysts in protecting cells from PN-mediated
 damage, a quantity of peroxynitrite(in 50 mM NaOH) determined to cause
 maximal injury(100%) was added as an exogenous pulse to each well of cells
 in the presence or absence of catalyst. The NaOH vehicle was not toxic by
 itself.
 Cells(RAW 264.7 cells or P815 mastocytoma cells; American Type Culture
 Collection, Rockville, Md.) were plated to confluence on 96-well tissue
 culture plates. Each well is washed twice with Dulbecco's phosphate
 buffered saline(DPBS; GIBCO BRL, Grand Island, N.Y.) to remove protein and
 other serum components which might react with the exogenous peroxynitrite.
 To each well is then added 200 .mu.l of DPBS. PN is next placed into
 separate wells at increasing concentrations and cell viability monitored.
 The dose at which maximal cell death is attained is then utilized for the
 catalyst protection assessment.
 Phosphate-buffered saline (200 uL) containing increasing concentrations of
 catalyst is next placed into individual wells of cells. The maximal dose
 of PN is subsequently administered to all wells of cells. After 15
 minutes, the medium is removed from each well and the cells are either
 allowed to recover overnight in Earles minimum essential medium without
 phenol red and supplemented with 10% fetal bovine serum or alternatively
 the plate of cells is assayed that day for mitochondrial integrity using
 the Alamar Blue viability assay(Alamar Biosciences, Inc.; Sacramento,
 Calif.). In either case, cells are incubated at 37.degree. C. in 5%
 CO.sub.2.
 Cell injury is measured as follows. Briefly, 10% Alamar Blue(v/v) in Earles
 MEM with 10% FBS is added to each well of cells for 1-2h. Cell metabolism
 of the dye generates a fluorescent product which is directly related to
 the number of viable cells. Moreover, the production of the fluorescent
 metabolite is linear for over 2 h. The amount of fluorescent product in
 100 .mu.l of conditioned medium from each well of cells is then measured
 with an IDEXX fluorescent plate reader (gain setting of 1%) at an emission
 wavelength of 575 nm after exciting at 545 nm. Viability is either given
 as absolute fluorescent units or as a percent of the value obtained for
 untreated cells(100%).
 As can be seen in FIG. 8, both Fe- and Ni- coordinated catalysts were able
 to protect the murine monocyte-macrophage line RAW 264.7; in this
 experiment PN was added at a dose causing a 50% decrease in cell
 viability.
 Comparison of increasing PN doses on RAW and P815 cells showed no evidence
 for a differential susceptibility to peroxynitrite-mediated injury(data
 not shown). However, as shown in FIG. 9, there is a significant protection
 of cells by Fe-TMPyP, FeTMPS, and FeTPPS while H.sub.2 TMPyP and ZnTMPyP
 were relatively ineffective (data not shown), a result consistent with
 their lack of catalytic potency. Addition of catalyst after PN was unable
 to rescue the cells from injury (data not shown) indicating the ability of
 the catalysts to protect cells directly from oxidative damage due to PN.
 Example 17
 In vivo Evaluation:
 Carrageenan-induced paw edema. The effects of peroxynitrite catalysts in
 vivo were initially tested on the carrageenan-induced paw edema. The
 choice of using this in vivo model of inflammation was based on the
 knowledge that 1) the inflammatory response is blocked by NOS inhibitors
 and 2) by superoxide dismutase (SOD). This indicates the participation of
 both NO and of O.sub.2-. Male Sprague Dawley rats were purchased from
 Harlan Sprague-Dawley (Indianapolis, Ind.). Male Sprague Dawley rats
 (175-200 g) received a subplantar injection in the right hind paw of
 carrageenan (0.1 ml of a 1% suspension in 0.85% saline). Paw volume was
 measured by a plethysmometer immediately before the injection of
 carrageenan and then at hourly intervals from 1 to 6 h. Edema was
 expressed as the increase in paw volume (in ml) measured after carrageenan
 injection compared to the pre-injection value for individual animals.
 Rats were given a bolus i.v. injection of active or inactive peroxynitrite
 catalysts 1 hour after the intraplantar injection of carrageenan; paw
 swelling was assessed thereafter every hour for up to 6 h. The relative %
 inhibition obtained with these agents is summarized in Table 2. Under
 these experimental conditions the inactive peroxynitrite catalysts H.sub.2
 TMPS, ZnTMPyP or MnTPPS (all given at 30 mg/kg) or FeCl.sub.3 (5 mg/kg,
 n=6) failed to inhibit edema formation.
 TABLE 2
 % Inhibition of Paw Edema bv Peroxynitrite Decomposition Catalysts
 Time (h) Post Carrageenan
 Compound Dose(mg/kg) 1 2 3 4 5 6
 FeTMPS 3 0 42 47 47 33 33
 10 0 61 60 53 53 47
 30 0 85 80 80 80 81
 FeTMPy 3 0 9 10 17 6 0
 10 6 13 11 28 21 2
 30 0 44 43 50 32 32
 FeTPPS 3 0 29 20 20 19 5
 10 0 17 20 23 19 20
 30 0 27 25 30 34 33
 ZnTMPS 30 0 0 0 0 0 0
 H.sub.2 TMPS 30 0 0 0 0 0 0
 MnTMPS 30 0 0 0 0 0 0
 Results are expressed as % inhibition of paw edema when compared to values
 obtained in control rats at the same time points. Each point is the mean
 .+-. s.e.m for n = 6 animals.
 Induction of intestinal damage by endotoxin in the rat: Multiple organ
 failure syndrome (MOFS) that develops following the septic attack is in
 most cases fatal. The "motor" of MOFS is the gastrointestinal tract, in
 particular the small intestine. Extensive ischaemia may be found in the
 intestinal mucosa due to profound vasoconstriction. Ischaemia and hypoxia
 result in mucous lesions, found both in animals (rat, cat, dogs) and
 humans. The origin of the mucous lesion is hypoxia. During reperfusion
 (e.g, after the initial severe vasoconstriction), O.sub.2- may be
 liberated and play an important role in the pathogensis of mucous lesions
 in the GI tract. Intestinal damage that results from shock induced by
 sphlanchnic artery occlusion is prevented by superoxide dismutase and LPS
 induced intestinal inflammation is inhibited by non-selective inhibitors
 of the nitric oxide pathway (Boughton-Smith, N. K et al., 1993). There is
 now substantial experimental and clinical evidence that suggests that
 excessive NO production has an important pathological role in the
 hypotension, hyporesponsiveness to vasoconstictors and the cardiovascular
 collapse associated with septic shock. Furthermore, nitric oxide synthase
 inhibitors prevent against the intestinal damage caused by endotoxin. We
 have developed a model of intestinal injury in rats by endotoxin and
 assessed the effects of therapeutic administration of peroxynitrite
 catalysts.
 Intestinal vascular permeability was determined as the leakage into the
 jejunal tissue of [.sup.125 I]-labelled bovine serum albumin ([.sup.125
 ]-BSA) administered intravenously (0.5 ml; 0.5 .mu.Ci) together with
 either LPS (3 mg/kg, serotype O111:B4) or isotonic saline. At 4 h after
 LPS administration, segments of jejunal tissue were ligated and removed.
 The intestinal tissues were rapidly washed, blotted dry and weighed. Blood
 (0.5 ml) was collected into tubes containing tri-sodium citrate (0.318%
 final concentration) and plasma prepared by centrifugation (10,000
 g.times.10 min). The {.sup.125 I]-BSA content in segments of whole tissue
 and in aliquots of plasma (100 .mu.l) was determined in a gamma counter.
 The total content of plasma in the intestinal tissues was expressed as
 .mu.l/g tissue. Changes in intravascular volume in the intestinal tissue
 was determined in an additional group of rats by administering ([.sup.125
 I]-BSA) intravenously 2 min before removal of the jejunum. The tissue and
 plasma content of radiolabel was determined and intravascular volume
 expressed as .mu.l/g tissue. This value was substracted from that obtained
 in the plasma leakage studies to obtain a measure of the intestinal plasma
 albumin leakage. After LPS administration (4 h), there was a significant
 (P&lt;0.01) increase in the plasma leakage (from 77.+-.10 to 224.+-.18
 .mu.l/g tissue, n=8). Administration of FeTMPS or FeTMPyP (30 mg/kg, i.v,
 n=4), 3 h after LPS injection, caused a reduction in radiolabelled albumin
 leakage determined 1 h later, as shown in FIG. 10. In contrast,
 administration of the inactive peroxynitrite catalyst ZnTMPyP (30 mg/kg,
 i.v, n=4), 3 h after LPS injection, did not inhibit radiolabelled albumin
 leakage determined 1 h later (FIG. 10). This data was supported by
 histological examination of the jejunal tissues. When compared to saline
 treated rats, LPS evoked profound jejunal damage with severe disruption of
 plicae and villi. LPS-induced damage was less severe in jejunums taken
 from rats treated with FeTMPS or FeTMPyP (30 mg/kg, i.v.).
 Thus, the compounds which are compounds or complexes of the present
 invention are novel and can be utilized to treat numerous inflammatory
 disease states and disorders. For example, reperfusion injury to an
 ischemic organ, e.g., reperfusion injury to the ischemic myocardium,
 inflammatory bowel disease, rheumatoid arthritis, osteoarthritis,
 hypertension, psoriasis, organ transplant rejections, organ preservation,
 impotence, radiation-induced injury, asthma, atherosclerosis, thrombosis,
 platelet aggregation, side effects from drug treatment of cancer
 metastasis, influenza, stroke, burns, trauma, acute pancreatitis,
 pyelonephritis, hepatitis, autoimmune diseases, insulin-dependent diabetes
 mellitus, disseminated intravascular coagulation, fatty embolism, adult
 and infantile respiratory distress, and hemorrhages in neonates.
 Patients receiving IL-2 therapy often develop potentially life-threatening
 side effects that include fever, chills, hypotension, capillary leak
 syndrome, as well as evidence of multiple organ dysfunction, specifically
 including renal insufficiency and cholestatic jaundice. IL-2 induces a
 complex network of cytokines that include tumor necrosis factor,
 interleukin 1 and 6. Therefore, IL-2-treated patients resemble patients
 with endotoxemia (hypotension, elevated TNF levels, elevated cytokine
 levels etc). Some of these induce release of free radicals as well as
 inducing iNOS with subsequent release of NO. A recent paper shows that
 iNOS is induced in patients that receive IL-2 for treatment of renal cell
 carcinoma and malignant melanoma (Hibbs, J. B. et al., Evidence for
 cytokine-inducible nitric oxide synthesis from L-arginine in patients
 receiving interleukin-2 therapy. J. Clin. Invest. Vol 89, 867-877).
 Activity of the compounds or complexes of the present invention for
 protecting superoxide dismutase can be demonstrated using the stopped-flow
 kinetic described above. Stopped-flow kinetic analysis is an accurate and
 direct method for quantitatively monitoring the decay rates of
 peroxynitrite in water. The stopped-flow kinetic analysis is suitable for
 screening complexes for catalytic peroxynitrite decomposition activity and
 active complexes of the present invention, as identified by stopped-flow
 analysis, are shown to correlate to treating the above disease states and
 disorders.
 In other words, the present invention is for the methods and compositions
 for the treatment of a disease or condition advantageously affected by
 decomposition of peroxynitrite which is accelerated over a natural
 background rate of decay, preferably in humans suffering from such disease
 or condition, which comprises administering a metal complex, in dosage
 unit form, of accelerated-rate-effective amounts for decomposing
 peroxynitrite preferably wherein the metal complex is as defined above.
 Such methods or compositions accomplish the treatment of these diseases
 without disadvantageously affecting normal biologically advantageous
 mechanisms.
 Total daily dose administered to a host in single or divided doses may be
 in amounts, for example, from about 1 to about 100 mg/kg body weight daily
 and more usually about 3 to 30 mg/kg. Dosage unit compositions may contain
 such amounts of submultiples thereof to make up the daily dose. The number
 of submultiples is preferably about one to three times per day of about 30
 mg/kg per unit dosage form. The serum concentrations of the doses are
 about 15 .mu.M to 1.5 mM with preferred ranges of 3 to 300 .mu.M.
 The amount of active ingredient that may be combined with the carrier
 materials to produce a single dosage form will vary depending upon the
 host treated and the particular mode of administration.
 The dosage regimen for treating a disease condition with the compounds
 and/or compositions of this invention is selected in accordance with a
 variety of factors, including the type, age, weight, sex, diet and medical
 condition of the patient, the severity of the disease, the route of
 administration, pharmacological considerations such as the activity,
 efficacy, pharmacokinetic and toxicology profiles of the particular
 compound employed, whether a drug delivery system is utilized and whether
 the compound is administered as part of a drug combination. Thus, the
 dosage regimen actually employed may vary widely and therefore may deviate
 from the preferred dosage regimen set forth above.
 The compounds of the present invention may be administered orally,
 parenterally, by inhalation spray, rectally, or topically in dosage unit
 formulations containing conventional nontoxic pharmaceutically acceptable
 carriers, adjuvants, and vehicles as desired. Topical administration may
 also involve the use of transdermal administration such as transdermal
 patches or iontophoresis devices. The term parenteral as used herein
 includes subcutaneous injections, intravenous, intramuscular, intrasternal
 injection, or infusion techniques.
 Injectable preparations, for example, sterile injectable aqueous or
 oleaginous suspensions may be formulated according to the known art using
 suitable dispersing or wetting agents and suspending agents. The sterile
 injectable preparation may also be a sterile injectable solution or
 suspension in a nontoxic parenterally acceptable diluent or solvent, for
 example, as a solution in 1,3-butanediol. Among the acceptable vehicles
 and solvents that may be employed are water, Ringer's solution, and
 isotonic sodium chloride solution. In addition, sterile, fixed oils are
 conventionally employed as a solvent or suspending medium. For this
 purpose any bland fixed oil may be employed including synthetic mono- or
 diglycerides. In addition, fatty acids such as oleic acid find use in the
 preparation of injectables.
 Suppositories for rectal administration of the drug can be prepared by
 mixing the drug with a suitable nonirritating excipient such as cocoa
 butter and polyethylene glycols which are solid at ordinary temperatures
 but liquid at the rectal temperature and will therefore melt in the rectum
 and release the drug.
 Solid dosage forms for oral administration may include capsules, tablets,
 pills, powders, granules and gels. In such solid dosage forms, the active
 compound may be admixed with at least one inert diluent such as sucrose
 lactose or starch. Such dosage forms may also comprise, as in normal
 practice, additional substances other than inert diluents, e.g.,
 lubricating agents such as magnesium stearate. In the case of capsules,
 tablets, and pills, the dosage forms may also comprise buffering agents.
 Tablets and pills can additionally be prepared with enteric coatings.
 Liquid dosage forms for oral administration may include pharmaceutically
 acceptable emulsions, solutions, suspensions, syrups, and elixirs
 containing inert diluents commonly used in the art, such as water. Such
 compositions may also comprise adjuvants, such as wetting agents,
 emulsifying and suspending agents, and sweetening, flavoring, and
 perfuming agents.
 While the compounds of the invention can be administered as the sole active
 pharmaceutical agent, they can also be used in combination with one or
 more compounds of the present invention or with one or more compounds
 which are known to be effective against the specific disease state that
 one is targeting for treatment.