MXPA98001322A - Bioconjugados de complejos de manganeso de ligandos macrociclicos containing nitrogen, effective as catalysts to dismute superox - Google Patents

Abstract

Bioconjugates of imitators of the superoxide dismutase (SOD) of low molecular weight represented by the formula (I): characterized in that R, R ', R1, R'1, R2, R'2, R3, R'3, R4, R '4, R5, R'5, R6, R'6, R7, R'7, R8, R'8, R9, R'9, X, Y, Z and n are as defined in the present invention, useful as therapeutic agents for conditions and disorders due to inflammatory disease, such as ischemic reperfusion injury, fulminating crisis, atherosclerosis, and all other tissue damage or injury conditions induced by oxidant

Description

BIOCONJUGADOS DE COMPLEJOS DE MANGANESO DE LIGANDOS MACROCICLICOS CONTAIN NITROGEN. EFFECTS AS CATALYSTS TO DISMOKE SUPEROXIDE BACKGROUND OF THE INVENTION The present invention relates to effective compounds as catalysts for dissolving superoxide. This invention relates to manganese (II) or manganese (III) complexes of fifteen-member nitrogen-containing macrocyclic ligands, which catalytically dissolve superoxide. In another aspect, this invention relates to manganese complexes of fifteen-member macrocyclic ligands containing nitrogen, which are conjugated to a selection biomolecule.

RELATED TECHNIQUE The enzyme superoxide dismutase catalyzes the conversion of superoxide to oxygen and hydrogen peroxide according to equation (1) (hereinafter called dismutation). 02 - + 02 - + 2H + 02 + H2O2 (1) It is believed that reactive oxygen metabolites derived from superoxide contribute to tissue pathology in a number of inflammatory diseases and disorders, such as reperfusion injury in the ischemic myocardium, disease inflammatory bowel, rheumatoid arthritis, osteoarthritis, atherosclerosis, hypertension, metastasis, psoriasis, rejection of transplanted organs, radiation-induced injuries, asthma, influenza, attacks, burns and traumas. See, for example, Bulkley, G.B. , Reactive oxygen metabolites and reperfusion injury: aberrant triggering of reticuloendothelial function, The Lancet, Vol. 344, pp. 934-36, October 1, 1994; Grisham, M.B., Oxidants and free radicals in inflammatory bowel disease, The Lancet, Vol. 344, p. 859-861, September 24, 1994; Cross, CE. and others, Reactive oxygen species and the lung, The Lancet, Vol. 344, pp. 930-33, October 1, 1994; Jenner, P., Oxidative damage in neurodegenerative disease, The Lancet, Vol. 344, pp. 796-798, September 17, 1994; Cerutti, P.A., Oxy-radicals and cancer, The Lancet, Vol. 344, pp. 862-863, September 24, 1994 Simic, M. G., and others, Oxygen Radicáis in Biology and Medicine, Basic Life Sciences, Vol. 49, Plenum Press, New York and London, 1988; -eiss J. Cell. Biochem., 1991 Suppl. 15C, 216 Abstract C110 (1991); Petkau, A., Cancer Treat. Rev. 13, 17 (1986); McCord, J. Free Radicáis Biol. Med., 2, 307 (1986); and Bannister, J.V. and others, Crit. Rev. Biochem., 22 111 (1987). The previously identified references of The Lancet teach the nexus between free radicals derived from superoxide and a variety of diseases. In particular, the Bulkley and Grisham references specifically teach that there is a link between the dismutation of superoxide and the treatment of the final disease. It is also known that superoxide is involved in the breakdown of the vascular relaxant factor derived from the endothelium (EDRF), which has been identified as nitric oxide (NO), and that the EDRF is protected from degradation by superoxide disutase. This suggests a central role for activated oxygen species derived from superoxide in the pathogenesis of vasospasm, thrombosis and atherosclerosis. See, for example, Grygle ski, R.J. and others, "Superoxide Anion is Involved in the Breakthrough of Endothelium-derived Vascular Relaxing Factor", Nature, Vol. 320, pp. 454-56 (1986) and Plamer, R.M.J. and others, "Nitric Oxide Relay Accounts for the Biological Activity of Endothelium Derived Relaxing Factor", Nature. Vol. 327, pp. 523-26 (1987). Clinical tests and animal studies with natural, recombinant and modified superoxide dismutase enzymes have been completed or are carried out to demonstrate the therapeutic efficacy of reducing superoxide levels in the aforementioned disease states. However, numerous problems have been created with the use of enzymes as potential therapeutic agents, including lack of oral activity, short half-lives in vivo, ingenicity with non-human-derived enzymes and deficient tissue distribution. The manganese complexes of the fifteen-member nitrogen-containing macrocyclic ligands that are low molecular weight replicates of superoxide dismutase (SOD) are useful as therapeutic agents and avoid many of the problems associated with SOD enzymes. Nevertheless, it would be desirable to be able to direct SOD replicas to a desired target in the body, where the compound can be concentrated for an optimal effect. Without a way of making the "selection" compounds, increased doses are sometimes needed in order to obtain an effective concentration at the site of interest. Said increased doses may sometimes result in undesirable side effects in the patient. It has now been discovered that the macrocycles or manganese complexes of the present invention can be fixed, ie conjugated, to one or more selection biomolecule (s) by means of a linking group to form a complex conjugate of biomolecule selection-macrocycle or of selection-manganese biomolecule.

BRIEF DESCRIPTION OF THE INVENTION It is an object of the invention to provide bioconjugates of manganese (II) or manganese (III) complexes of fifteen-member nitrogen-containing macrocyclic ligands that are low molecular weight replicates of superoxide dismutase (SOD), which are useful as therapeutic agents for states or inflammatory disease disorders that are mediated, at least in part, by superoxide. It is a further object of the invention to provide bioconjugates of manganese (II) complexes of fifteen-member, nitrogen-containing macrocyclic ligands, which are useful as contrast agents for magnetic resonance imaging (MRI) that have kinetic stability improved, improved oxidative stability and improved hydrogen bond. It is also a further object of the invention to provide bioconjugates of manganese complexes of fifteen-member macrocyclic ligands containing nitrogen that can be directed to a specific site in the body. According to the invention, biconjungados of complexes of manganese (II) or manganese (III) of macrocyclic ligands of fifteen members containing nitrogen are provided, in which (1) one to five of the groups "R" are fixed to biomolecules by means of a linking group, (2) one of X, Y and Z is fixed to a biomolecule by means of a linking group or (3) one to five of the groups "R" and one of X, Y and Z are fixed to biomolecules by means of a linking group; and the biomolecules are independently selected from the group consisting of steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and receptor substrates. of enzyme, and the linking group is derived from a substituent attached to the group "R", or X, Y and Z, which is reactive with the biomolecule and is selected from the group consisting of -NH2, -NHRio, -SH, -OH, -COOH, -COOR, -CONH2, -NCO, -NCS, -COOX ", alkenyl, alkynyl, halide, tosylate, esylate, tresylate, triflate and phenol, wherein Rio is alkyl, aryl or alkylaryl and X" It is a halide.

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to bioconjugates of manganese (II) or manganese (III) complexes of fifteen-member nitrogen-containing macrocyclic ligands, which catalyze the conversion of superoxide to oxygen peroxide and hydrogen. These complexes can be represented by the formula: where R, R ', Ri, Ri', R2, R2 ', R3, R3', R ', R.,', Rs, Rs ', Re, Re', R7, R? ', Rs, Rβ', R9 and 9 'independently represents hydronogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, alkylcycloalkyl, alkenylcycloalkyl, alkylcycloalkenyl, alkenylcycloalkenyl, heterocyclic, aryl and aralkyl radicals, and radicals attached to the α-amino acid carbon; or R or R'i and R2 or '2, R3 or R'3 and R4 or R' 4, Rs or R's and Re or R'ß, 7 or R'7 and Rß or R'ß, and R9 or R '9 and R or R' together with the carbon atoms to which they are attached, independently form a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms; or R or R 'and Ri or R'i, R2 or R'2 and R3 or R'3, * or R' 4 and Rs or R'd. d or R's and R7 or R'7, and Rs or R'ß R9? R'9 together with the carbon atoms to which they are attached, independently form a nitrogen-containing heterocycle having from 2 to 20 carbon atoms; provided that when the nitrogen-containing heterocycle is an aromatic heterocycle that does not contain a hydrogen bound to nitrogen, the hydrogen bound to nitrogen in said formula, in which the nitrogen is also in the macrocycle, and the R groups attached to the The same carbon atoms of the macrocycle are absent; and combinations thereof; and wherein (1) one to five of the "R" groups are fixed to biomolecules by means of a linker group, (2) one of X, Y and Z is attached to a biomolecule by means of a linking group or (3) ) one to five of the "R" groups and one of X, Y and Z are fixed to biomolecules by means of a linker group; and the biomolecules are independently selected from the group consisting of steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and receptor substrates of enzyme, and the linker group is derived from a substituent attached to the group "R" or X, Y and Z, which is reactive with the biomolecule and is selected from the group consisting of -NH2, -NHR10, -SH, - OH, -COOH, COOR10, -CONH2, -NCO, -NCS, -COOX ", alkenyl, alkynyl, halide, tosylate, mesylate, tresylate, triflate and phenol, wherein Rio is alkyl, aryl or alkylaryl and X" is a halide. X, Y and Z represent suitable charge neutralizing ligands or anions which are derived from any monodentate or polydentate ligand or ligand system or the corresponding anion thereof (eg benzoic acid or benzoate anion, phenol or phenoxide anion). , alcohol or alkoxide anion). X, Y and Z are independently selected from the group consisting of halide, oxo, water, 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 such as acetic acid, trifluoroacetic acid, oxalic acid), aryl carboxylic acid (such as benzoic acid and phthalic acid) o), urea, alkyl urea, aryl urea, alkyl aryl urea, thiourea, alkyl thiourea, aryl thiourea, alkyl aryl thiourea, sulfate, sulphite, bisulfate, bisulfite, thiosulfate, thiosulphite, hydrosulfite, alkyl phosphine, aryl phosphine, alkyl oxide phosphine, aryl phosphine oxide, alkyl aryl phosphine oxide, alkyl phosphine sulfide, aryl phosphine sulfide, alkyl aryl phosphine sulfide, alkyl phosphonic acid, aryl phosphonic acid, phenyl phosphinic acid, aryl phosphinic acid, alkyl phosphino acid, acid phosphinate, phosphate, thiophosphate, phosphite, pyrophosphite, triphosphate, hydrogen phosphate, dihydrogen phosphate, alkyl guanidino, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alky thiocarbamate, aryl thiocarbatoxide, alkyl aryl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkyl aryl dithiocarb ate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalogen anganate, tetrafluoroborate, hexafluorophosphate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetra alkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxyamic acid, thiotosylate and ion exchange resin anions systems in which one or more of X, Y and Z are independently fixed to one or more of the "R" groups, wherein n is 0 or 1. Preferred ligands from which X, Y and Z are selected include halide, organic acid, and nitrate and bicarbonate anions. The linker groups, also called "linker" here, are derived from the specific functional groups attached to the "R" or X, Y and Z groups, and function to link the biomolecule to the "R", or X, Y groups. and Z. The functional groups are selected from the group consisting of -NH2, -NHR10, -SH, -OH, -C00H, COORio, -CONH2, -NCO, -NCS, -C00X, "alkenyl, alkynyl, halide, tosylate. , mesylate, tresylate, triflate and phenol, in which Rio is alkyl, aryl or alkaryl and X "is a halogenide Currently, the preferred alkenyl group is ethenyl and the preferred alkynyl group is ethynyl The functional groups in the" R "or X, Y and Z groups are reactive with the biomolecule, i.e. , reactive with a functional group on steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors, enzyme receptor substrates, and other biomolecules of selection of interest.When the functional group fixed to the "R" or X, Y and Z groups reacts with the biomolecule, the functional group is modified and it is this derivative group which is the linker. a functional group -NH2 attached to a group "R" is reacted with a steroid such as in example 1, the linker is -NH- The exact structure of the specific linking groups will be readily apparent to that group. Those skilled in the art will depend on the specific functional group and the selected biomolecule. Specific reaction conditions for reacting a functional group attached to the "R" or X, Y and Z groups with a biomolecule will be readily apparent to those skilled in the art. The functional group useful for forming the linker, defined herein as a "linker precursor", may be present in the "R" groups at the time the macrocycle is prepared or may be added or modified after the preparation of the macrocycle or complex. of manganese itself. Similarly, the linker precursor may be present in an axial ligand, i.e. X, Y or Z, when the manganese complex is prepared, or an exchange reaction of the axial ligands is conducted to exchange the axial ligands present in the complex. manganese. The macrocycle of the present invention can be complexed with manganese either before or after conjugation with the selection biomolecule depending on the specific biomolecule used. The conjugate of the macrocyclic complex and the selection biomolecule is defined herein as a "bioconjugate". The selection of drugs is well known to those skilled in the art. See, for example, J. A. Katzenellenbogen et al., Journal of Nuclear Medicine. Vol. 33, No. 4, 1992, 558, and J.A. Katzenellenbogen and others, Bioconjugate Chemistru. 1991, 2, 353. Selection agents are typically biomolecules. The biomolecules of the invention are biologically active molecules that are site-specific, that is, known to be concentrated in the particular organ or tissue of interest. The biomolecules are selected to direct the tissue distribution of the bioconjugate by binding to the receptor, membrane association, membrane solubility and the like. These biomolecules include, for example, steroids, carbohydrates, (including monosaccharides, disaccharides and polysaccharides), fatty acids, amino acids, peptides, proteins, antibodies (including polyclonal and monoclonal and fragments thereof), vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and enzyme receptor substrates. Biomolecules also include those biomolecules which are combinations of the above biomolecules, such as a combination of a steroid with a carbohydrate, v. gr. , digitonin. Particular biomolecules that can be used to select a desired organ or tissue are known in the art or will be readily apparent to those skilled in the art. The biomolecules of the invention are commercially available or can be easily prepared by one skilled in the art using conventional methods. It is currently preferred that a maximum of an "R" group attached to the carbon atoms located between the nitrogen atoms in the macrocycle have a biomolecule fixed by means of a linker. further, the preferred compounds are those having one to five, most preferably one to two of the "R" groups attached to biomolecules and none of X, Y and Z attached to a biomolecule, or those having one of X, Y and Z fixed to a biomolecule and none of the "R" groups fixed to a biomolecule. Currently, the preferred compounds are those in which at least one, most preferably at least two of the "R" groups, in addition to the "R" groups that are fixed to a biomolecule, represent alkyl, cycloalkylalkyl and aralkyl radicals and the remaining "R" groups not fixed to a biomolecule represent hydrogen, a saturated, partially saturated or unsaturated cyclic, or a heterocycle containing nitrogen. Other groups of preferred compounds are those in which at least one, preferably two, Ri or R'i and R2 or R'2, R3 or R'3 and R «or R'4, Rs or R's and Rß or R'ß, R7 or R'7 and Rß or R'ß and R9 or R'9 and R or R 'together with the carbon atoms to which they are attached represent a saturated, partially saturated or unsaturated cyclic having 3 to 20 carbon atoms and the remaining "R" groups in addition to the "R" groups which are fixed to a biomolecule by means of a linker are hydrogen, nitrogen-containing heterocycles or alkyl groups, and those in which at least one, preferably two of Ri or R'i and R2 or R'2, R3 or R'3 and Rt, or R 'Ü, Rs or R's and Rβ or R'ß, R7 or R'7 and Rβ or R'ß, and R9 or R'9 together with the carbon atoms which are attached are bonded to form a nitrogen containing heterocycle having from 2 to 20 carbon atoms and the remaining "R" groups in addition to the "R" groups which are fixed in a biomolecule by means of a linker are independently selected from hydrogen, saturated cyclic, partially saturated or unsaturated or alkyl groups. As used herein, the "R" groups mean all groups R attached to the carbon atoms of the macrocycle, ie, R, R ', Ri, R'i, R2, R'2, R3, R' 3, Ri,, R '?, Re, R's, Rß, R'ß, R7, R'7, Re, R'ß, R9 and R'9. Another embodiment of the invention in a pharmaceutical composition in unit dosage form useful for dismutating superoxide comprising (a) a therapeutically or prophylactically effective amount of a complex as described above, and (b) a nontoxic carrier, adjuvant or vehicle and pharmaceutically acceptable. The commonly accepted mechanism of action of manganese-based SOD enzymes includes the cyclization of the manganese center between the two oxidation states (II, III). See J. V. Bannister, W.H. Bannister, and G. Rotilio, Crit.

Rev. Biochem., 22, 111-180 (1987). 1) Mn (II) + HO2 > Mn (III) + H02 2) Mn (III) + 02 - > Mn (II) + O2 The formal oxide-reduction potentials for the O2 / O2- and HO2 / H2O2 pairs at pH = 7 are -0.33 v and 0.87 v, respectively. See A. E.G. Cass, in Metalloproteins: Part 1, Metal Proteins with Redox Roles, ed. P. Harrison, P. 121. Verlag Chemie (Weinheim, GDR) (1985). For the mechanism described above, these potentials require that a putative SOS catalyst be able to rapidly undergo oxidation state changes in the range of -0.33 v to 0.87 v. The complexes derived from Mn (II) and the general class of ligands [15] C-substituted aneNs described herein have all been characterized using cyclic voltammetry to measure their oxide-reduction potential. The C-substituted complexes described herein have reversible oxidations of about +0.7 v (SHE). The coulometry shows that this oxidation is a one-electron procedure; namely, it is the oxidation of the Mn (II) complex to the Mn (III) complex. In this way, for these complexes to function as SOD catalysts, the oxidation state of Mn (III) is included in the catalytic cycle. This means that the Mn (III) complexes of all these ligands are equally competent as SOD catalysts, since it does not matter which form of (Mn (II) or Mn (III)) is present when the superoxide is present because the superoxide will simply reduce Mn (III) to Mn (II) by releasing oxygen. As used 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, which optionally carries one or more substituents selected from (1) -NR30R31, wherein R3 and R31 are independently selected from hydrogen, alkyl, aryl or aralkyl; or R 30 is hydrogen, alkyl, aryl or aralkyl and R 31 is selected from the group consisting of -NR 32 R 33, -OH, -OR 3, 0 S 0 0 X '- C - Z', -C - Z ", - Í-R35, -I-R36, -S-R37, and -P- € 0R3ß) (0R39); wherein R32 and R33 are independently hydrogen, alkyl, aryl or acyl, R34 is alkyl, aryl or alkaryl, Z 'is hydrogen, alkyl, aryl, alkaryl, -OR34, -SR34 or -NR40R / .1, wherein R40 and R41 are independently selected from hydrogen, alkyl, aryl or alkaryl, Z "is alkyl, aryl, alkaryl, - OR34, -SR34 or -NR40R41, R35 is alkyl, aryl, -OR34, or -NR40R41, R3β is alkyl, aryl or -NR40R41, R37 is alkyl, aryl or alkaryl, X 'is oxygen or sulfur and R38 and R39 are selected independently of hydrogen, alkyl or aryl; (2) -SR42 wherein R42 is hydrogen, alkyl, aryl, alkaryl, -SR34, -NR32R33, X '0 0 ~ C-Z ", -4-R" 3, or -P-eA) (B) wherein R43 is -OH, -OR34 or -NR32R33, A and B are independently -OR34, -SR34 or -NR-32R33 wherein x is 1 or 2, and R44 is halide, alkyl, aryl, alkaryl, -OH, -OR34, -SR34 or -NR32R33; (4) -OR45 wherein R45 is hydrogen, alkyl, aryl, alkaryl, -NR32R33, X'R «« 0 0 --C - Z ', ~ S (-0) x, -i-6D) (E ) or ~ P- € R34) (0R34); wherein D and E are independently -OR3-V or -NR32R33; X '-C-R «6 (5) wherein R« ß is halide, -OH, -SH, -OR34, -SR34 or -NR32R33; or (e) amine oxides of the formula ~ N + R 3 or R 3 A - provided that R 30 and 31 are not hydrogen; or (7) wherein F and G are independently -OH, -SH, -OR34, -SR34 or -NR32R33; or (8) -0 - (- (CH2) a-0) b -Rio wherein Rio is hydrogen or alkyl, and a and b are integers independently selected from 1 + 6; or (9) halogen, cyano, nitro or azido. The alkyl, aryl and alkaryl groups in the substituents of the alkyl groups defined above may contain an additional substituent, but preferably they are unsubstituted. Examples of such radicals include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isoyl, hexyl, octyl, nsyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl and eicosyl. The term "alkenyl", alone or in combination, means an alkyl radical having one or more double bonds. Examples of said alkenyl radicals include ethenyl, propenyl, 1-butenyl, cis-2-butenyl, trans-2-butenyl, iso-butylenyl, cis-2-pentenyl, trans-2-pentenyl, 3-methyl-1-butenyl, 2,3-dimethyl-2-butenyl, 1-pentenyl, 1-hexenyl, 1-octenyl, decenyl, dodecenyl, tet radecenyl, hexadecenyl, cis- and trans-9-octadecenyl, 1,3-pentadienyl, 2,4- pentadienyl, 2,3-pentadienyl, 1,3-hexadienyl, 2,4-hexadienyl, 5,8,11,14-eicosatetraenyl and 9,12,15-octadecatrienyl. The term "alkynyl", alone or in combination, means an alkyl radical having one or more triple bonds. Examples of such alkynyl groups include ethynyl, propynyl (propargyl), l-butynyl, 1-octynyl, 9-octadecynyl, 1,3-pentadinyl, 2,4-pentadinyl, 1,3-hexadinyl, and 2,4-hexadinyl. 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 said cycloalkyl radicals include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and perhydronaphthyl. The term "cycloalkylalkyl" means an alkyl radical as defined above, which is substituted by a cycloalkyl radical as defined above. Examples of cycloalkylalkyl radicals include: cyclohexylmethyl, cyclopentylmethyl, (4-isopropylcyclohexyl) methyl, (4-t-butyl-cyclohexyl) methyl, 3-cyclohexylpropyl, 2-cyclohexyl ethylpentyl, 3-cyclopentylmethylhexyl, 1- (4-neopentylcyclohexyl) methylhexyl and l- (4-isopropylcyclohexyl) -methylheptyl. The term "cycloalkylcycloalkyl" means a cycloalkyl radical as defined above, which is substituted by another cycloalkyl radical as defined above. Examples of cycloalkylcycloalkyl radicals include cyclohexylcyclopentyl and cyclohexylcyclohexyl. The term "cycloalkenyl", alone or in combination, means a cycloalkyl radical having one or more double bonds. Examples of cycloalkenyl radicals include cyclopentenyl, cyclohexenyl, cyclooctenyl, cyclopentadienyl, cyclohexadienyl and cyclooctadienyl. The term "cycloalkenylalkyl" means an alkyl radical as defined above, which is substituted by a cycloalkenyl radical as defined above. Examples of cycloalkenylalkyl radicals include 2-cyclohexen-1-ylmethyl, 1-cyclopenten-1-ylmethyl, 2- (1-cyclohexen-1-yl) ethyl, 3- (1-cyclopenten-1-yl) propyl, 1 - ( l-cyclohexen-1-ylmethyl) pentyl, 1- (1-cyclopenten-1-yl) hexyl, 6- (1-cyclohexen-1-yl) hexyl, 1- (1-cyclopentenyl-1-yl) nonyl and 1 - (1-cyclohexen-1-yl) nonyl. The terms "alkylcycloalkyl" and "alkenylcycloalkyl" mean a cycloalkyl radical as defined above, which is substituted by an alkyl or alkenyl radical as defined above. Examples of alkylcycloalkyl and alkenylcycloalkyl radicals include: 2-ethylcyclobutyl, 1-methylcyclopentyl, 1-hexylcyclopentyl, 1-methylcyclohexyl, 1- (9-octadecenyl) cyclopentyl and 1- (9-octadecenyl) cyclohexyl. The terms "alkylcycloalkenyl" and "alkenylcycloalkenyl" mean a cycloalkenyl radical as defined above, which is substituted by an alkyl or alkenyl radical as defined above. Examples of alkylcycloalkenyl and alkenylcycloalkenyl radicals include l-methyl-2-cyclopentenyl, l-hexyl-2-cyclopentenyl, l-ethyl-2-cyclohexenyl, l-butyl-2-cyclohexenyl, l- (9-octadecenyl) -2-cyclohexenyl and l- (2-pentenyl) -2-cyclohexenyl. The term "aryl", alone or in combination, means a phenyl or naphthyl radical optionally bearing 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-methoxyphenyl, 4- ( ter-butoxy) phenyl, 4-fluorophenyl, 4-chlorophenyl, 4-hydroxyphenyl, 1-naphthyl, 2-naphthyl and the like. The term "aralkyl", alone or in combination, means a radial alkyl or cycloalkyl as defined above, in which a hydrogen atom is replaced by an aryl radical as defined above, such as benzyl, 2-phenylethyl and the like. The term "heterocyclic" means ring structures that contain at least one other type of atom, besides carbon, in the ring. The most common of the other types of atoms include nitrogen, oxygen and sulfur. Examples of heterocyclics include the groups pi-Rididinyl, Piperidyl, I-aZolidinyl, Tetrahydrofuryl, Tetrahydrothienyl, Furyl, Thienyl, Pyridyl, Quinolyl, Isoquinolyl, Pyridazinyl, Pyrazinyl, Indolyl, Imidazolyl, Oxazolyl, Thiazolyl, Pyrazolyl, Pyridinyl, Benzoxadiazolyl, Benzothiadiazolyl, Triazolyl and tetrazolyl. The term "saturated, partially saturated or unsaturated cyclic" means fused ring structures in which two ring carbons are also part of the fifteen-membered acrocyclic ligand. The ring structure may contain 3 to 20 carbon atoms, preferably 5 to 10 carbon atoms, and may also contain one or more other types of atoms other than carbon. The most common of the other types of atoms include nitrogen, oxygen and sulfur. The ring structure may also contain more than one ring. The term "saturated, partially saturated or unsaturated ring structure" means a ring structure in which a ring carbon is also part of the fifteen-member acrocyclic ligand. The ring structure may contain 3 to 20, preferably 5 to 10 carbon atoms, and may also contain nitrogen, oxygen and / or sulfur atoms. The term "nitrogen-containing heterocycle" means ring structures in which two carbons and one ring nitrogen are also part of the fifteen-membered macrocyclic ligand. The ring structure may contain 2 to 20, preferably 4 to 10 carbon atoms, may be partially or completely unsaturated or saturated and may also contain nitrogen, oxygen and / or sulfur atoms in the ring portion that is also not part of the ligand macrocyclic of fifteen members. The term "organic acid anion" refers to carboxylic acid anions having from about 1 to about 18 carbon atoms. The term "halide" means chloride or bromide. The macrocyclic ligands useful in the complexes of the present invention can be prepared according to the general procedure shown in scheme A below. In this manner, an amino acid amide, which is the corresponding amine derivative of an oc-amino acid that occurs naturally or not naturally, is reduced to form the corresponding substituted ethylene diamine. Said amino acid amide may be the amide derivative of any of the well-known types of amino acids. Preferred amino acid amides are those represented by the formula: wherein R is derived from the forms or L's of the amino acids alanine, aspartic acid, arginine, asparagine, cysteine, glycine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, proline, phenylalanine, serine, tryptophan, threonine, tyrosine, valine and / or R groups of non-natural a-amino acids such as alkyl, ethyl, butyl, tert-butyl, cycloalkyl, phenyl, alkenyl, allyl, alkynyl, aryl, heteroaryl, polycycloalkyl, polycycloaryl, polycycloheteroaryl, imines , aminoalkyl, hydroxyalkyl, hydroxyl, phenol, amine oxides, thioalkyl, carboalkoxyalkyl, carboxylic acids and their derivatives, keto, ether, aldehyde, amine, nitrile, halogen, thiol, sulfoxide, sulfone, sulfonic acid, sulfide, disulfide, phosphonic acid , phosphinic acid, phosphine oxides, sulfonamides, amides, amino acids, peptides, proteins, carbohydrates, nucleic acids, fatty acids, lipids, nitro, hydroxylamines, hydroxyamic acids, thiocarbonyls , borates, boranes, boraza, silyl, siloxy, silaza and combinations thereof. Very preferred are those in which R represents hydrogen, alkyl, cycloalkylalkyl and aralkyl radicals. The diamine is then tosylated to produce the di-N-tosyl derivative which is reacted with a tris-N-tosylated di-0-tosylated triazaalkane diol to produce the corresponding substituted N-pentatosyl pentaazacycloalkane. The tosyl groups are then removed and the resulting compound is reacted with a manganese compound (II) under essentially anhydrous and anaerobic conditions to form the corresponding substituted manganese complex (I?) - pentaazacycloalkane. When charge neutralizing ligands or anions, ie, X, Y and Z are anions or ligands that can not be introduced directly from the manganese compound, the complex with those anions or ligands can be formed by conducting an exchange reaction with a complex that has been prepared by reacting the macrocycle with a manganese compound. The complexes of the present invention wherein R9 and R2 are alkyl, and R3, R'3, R < R ', R', R ', R', R ', R7, R'7, R', and R 'can be alkyl, arylalkyl or cycloalkylalkyl and R or R' and Ri or R'i together with the carbon to which they are fixed are likely to form a nitrogen-containing heterocycle, they can also be prepared according to the general procedure shown in scheme B below, using methods known in the art to prepare the manganese complex precursor (II) -pentaazabicyclo [12.3.1] octadecapentane. See, for example, Alexander and others, Inorg. Nucí Chem. Lett., 6, 445 (1970). In this way, a 2,6-diketopy ridine is condensed with triethylene tetraamine in the presence of a manganese compound (II) to produce the manganese (II) -pentaazabicyclo [12.3.1] octadecapentane complex. The complex of manganese (11) -pentaazabicyclo [12.3. lloctadecapentane is hydrogenated with platinum oxide at a pressure of .703-70.3 kg / cm2 to give the corresponding manganese (II) -pentaazabicyclo [12.3.1] octadecatriene complex. The macrocyclic ligands useful in the complexes of the present invention can also be prepared by means of the diacid dichloride route shown in Scheme C below. In this manner, a triazaalkane is tosylated in a suitable solvent system to produce the corresponding tris (N-tosyl) derivative. Said derivative is treated with a suitable base to produce the corresponding disulfonamide anion. The disulfonamide anion is dialkylated with a suitable electrophile to produce a derivative of a dicarboxylic acid. This derivative of a dicarboxylic acid is treated to produce the dicarboxylic acid, which is then treated with a suitable reagent to form the diacid dichloride. The desired neighborhood diamine is obtained in any of many ways. One form that is useful is the preparation from an aldehyde by reaction with cyanide in the presence of ammonium chloride followed by acid treatment to produce the alpha ammonium nitrile. The latter compound is reduced in the presence of acid and then treated with a suitable base to produce the vicinal diamine. The condensation of the diacid dichloride with the vicinal diamine in the presence of a suitable base forms the tris (tosyl) diamide macrocycle. The tosyl groups are removed and the amides are reduced and the resulting compound is reacted with a manganese compound (II) under essentially anhydrous and anaerobic conditions to form the corresponding substituted pentaazacycloalkane-manganese (II) complex. The vicinal diamines have been prepared by the route shown (known as the Strecker synthesis) and the vicinal diamines were purchased when commercially available. Any method of preparation of vicinal diamine can be used. The macrocyclic ligands useful in the complexes of the present invention can also be prepared by means of the pyridinediane route shown in Scheme D below. In this way, a polyamine such as a tetraaza compound, containing the primary amines is condensed with dimethyl 2,6-pi-dimeric dicarboxylate by heating in a suitable solvent, e.g., methanol, to produce a macrocycle incorporating the pyridine ring as the 2,6-dicarboxamide. The pyridine ring in the macrocycle is reduced to the corresponding piperidine ring in the macrocycle, and then the diamines are reduced and the resulting compound is reacted with a manganese compound (II) under essentially anhydrous and anaerobic conditions to form the complex of corresponding substituted pentaazacycloalkane-manganese (II). The macrocyclic ligands useful in the complexes of the present invention can also be prepared by the bis (halogenacetamide) route shown in Scheme E below. In this manner, a triazaalkane is tosylated in a suitable solvent to produce the corresponding tris (N-tosyl) derivative. Said derivative is treated with a suitable base to produce the corresponding disulfonamide anion. A bis (halogenoaceta ida), e.g., a bis (chloroacetamide) of a vicinal diamine is prepared by reacting the diamine with an excess of halogenoacetyl halide, e.g., chloroacetyl chloride, in the presence of a base. The disulfonamide anion of the tris (N-tosyl) triazaalkane is then reacted with the bis (chloroacetamide) of the diamine to produce the tris (N-tosyl) substituted diamide macrocycle. The tosyl groups are removed and the amides are reduced and the resulting compound is reacted with a manganese compound (II) under essentially anhydrous and anaerobic conditions to form the corresponding substituted pentaazacycloalkane-manganese (II) complex. The macrocyclic ligands useful in the complexes of the present invention, wherein Ri, R'i, R2, R'2 are derived from a diamino starting material and Rs, R's, R7, R'7 and R9, R'9 can be H or any functionality previously described, can be prepared according to the pseudopeptide method shown in scheme F below. A substituted 1,2-diaminoethane represented by the formula wherein Ri, R'i, R2, R'2 are the substituents on the adjacent carbon atoms in the macrocyclic ligand of the product as described above, it can be used in this method in combination with any amino acid. The diamine can be produced by any conventional method known to those skilled in the art. The R groups in the macrocycle derived from substituents on the a-carbon of α-amino acids, ie Rs, R's, R7, R'7, R9, and R'9, can be derived from the D or L forms of the amino acids alanine, aspartic acid, arginine, asparagine, cysteine, glycine, glutamine, glutamine, histidine, isoleucine, leucine, lysine, methionine, proline, phenylalanine, serine, tryptophan, threonine, tyrosine, valine and / or R groups of unnatural cx-amino acids such as alkyl, ethyl, butyl, tert-butyl, cycloalkyl, phenyl, alkenyl, allyl, alkynyl, aryl, heteroaryl, polycycloalkyl, polycycloaryl, polycycloheteroaryl, imines, aminoalkyl, hydroxyalkyl, hydroxyl, phenol, oxides of amine, thioalkyl, carboalkoxyalkyl, carboxylic acids and their derivatives, keto, ether, aldehyde, amine, nitrile, halogen, thiol, sulfoxide, sulfone, sulfonic acid, sulfide, disulfide, phosphonic acid, phosphinic acid, phosphine oxides, sulfonamides, amides , amino acids, peptides, proteins, carbohydrates, nucleic acids, fatty acids, lipids, nitro, hydroxylamines, hydroxyamic acids, thiocarbonyls, borates, boranes, boraza, silyl, siloxy, silaza and combinations thereof. As an example, 1,8-dihydroxy, 4,5-diaminooctane is monosylated and reacted with Boc anhydride to produce the N-Boc, N-tosyl differentiated derivative. The sulfonamide was alkylated with methyl bromoacetate using sodium hydride as the base and was saponified to the free acid. The N-tosylglycine containing diamine serves as a substitute for dipeptide in the normal synthesis of peptide in solution-phase. In this way, the doubling with a functionalized amino acid ester produces the corresponding pseudotripeptide. Two sequential cuts-copulations of TFA produce the pseudopentapeptide that can be deprotected in the N- and C- terminus in one step using HCI / AcOH. The DPPA-mediated cyclization followed by the reduction with LiAlH * or borane produces the corresponding macrocyclic binding. This ligand system is reacted with a manganese (II) compound, such as manganese (II) chloride under essentially anaerobic conditions to form the corresponding functionalized pentaazacycloalkane manganese (I?) Complex. When charge neutralizing ligands or anions, ie, X, Y, and Z are anions or ligands that can not be introduced directly from the magnesium compound, the complex with those anions or ligands can be formed by conducting an exchange reaction with a complex that has been prepared by reacting the macrocycle with a manganese compound. The acrocylic ligands useful in the complexes of the present invention, wherein Ri, R'i, R3, R'3 Rs, R's, R7, R'7, R9, and R'9 can be H or any functionality co or those described above, can be prepared according to the general peptide method shown in scheme G below. The R groups in the macrocycle derived from substituents on the α-carbon of o-amino acids, ie Ri, R'i, R3, R'3, Rs, R's, R7, R'7, R9, and R'9, are defined in scheme F. The process for preparing the cyclic peptide precursors from the corresponding linear peptides are the same or significant modifications of methods known in the art. See, for example, Veber, D.F. and others, J. Org. Chem., 44, 3101 (1979). The general method delineated in Scheme G below is an example using the sequential phase-solution solution of the linear pentapeptide functionalized from the N terminus to the C terminus. Alternatively, the reaction sequence for preparing the linear pentapeptide can be carried out by solid-phase preparation using methods known in the art. The reaction sequence can be conducted from the C terminus to the N terminus and by convergent scopes such as the doubling of di- and tri-peptides as needed. In this way, a Boc-protected amino acid is coupled with an amino acid ester using normal peptide coupling reagents. The new Boc-dipeptide ester is then saponified to the free acid, which is coupled again with another amino acid ester. The resulting Boc-tripeptide ester is saponified again and this method is continued until the free acid of the Boc-protected pentapeptide is prepared. The Boc protecting group is removed under normal conditions and the resulting pentapeptide or salt thereof is converted to the cyclic pentapeptide. The cyclic pentapeptide is then reduced to pentaazacyclopentadecane with lithium aluminum hydride or borane. The final ligand is then reacted with a manganese compound (II) under essentially anaerobic conditions to form the corresponding manganese (II) -pentaazacyclopentadecane complex. When charge-neutralizing ligands or anions, eg, X, Y and Z are anions or ligands that can not be introduced directly from the manganese compound, the complex with those anions or ligands can be formed by conducting a reaction of exchange with a complex that has been prepared by reacting the macrocycle with a manganese compound.

SCHEME A SCHEME B MnCl2 MeOH R02 H2. 7-03 kg / an2 MeOH, 100 ° C SCHEME C SCHEME D SCHEME E SCHEME F SCHEME F (cont.) SCHEME G NaOH, H20 CH3OH SCHEME 6 (cont.) NaOH H20 CH3OH The pentaazamac sprays of the present invention may 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 non-racemic mixtures thereof. The optical isomers can be obtained by resolution of the racemic mixtures in accordance with conventional procedures, for example, by formation of diastereomeric salts by treatment with an optically active acid. Examples of suitable acids are tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid and camphor sulfonic acid, then separating the mixture of diastereoisomers by crystallization, followed by release of the optically active bases from these salts. A different procedure for separating optical isomers is to use a chiral chromatography column optimally chosen to maximize the separation of the enantiomers. Yet another available method involves the synthesis of covalent diastereomeric molecules by reacting one or more secondary amine groups 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 release the enantiomerically pure ligand. In the same way, the optically active compounds of the invention can be obtained using optically active starting materials, such as natural amino acids. The compounds or complexes of the present invention are novel and can be used to treat numerous conditions and disorders of inflammatory disease. For example, reperfusion injury to an ischemic organ, eg, reperfusion injury to the ischemic myocardium, surgically induced ischemia, inflammatory bowel disease, rheumatoid arthritis, osteoarthritis, psoriasis, rejection to organ transplantation, radiation induced injury, injury and tissue damage induced by oxidants, atherosclerosis, thrombosis, platelet aggregation, fulminant crisis, acute pancreatitis, insulin-dependent diabetes mellitus, disseminated intravascular coagulation, fat embolism, respiratory distress in children and adults, metastasis and carcinogenesis. The activity of the compounds or complexes of the present invention to catalyze the dismutation of superoxide can be demonstrated using the flow kinetics impeded analysis technique as described in Riley, D.P., Rivers, W.J. and Weiss, R.H., "Stopped-Flo Kinetic Analysis for Monitoring Superoxide Decay in Aqueous Systems", Anal. Biochem., 196, 344-349 (1991). The analysis of the hindered flow kinetics is a direct and precise method to quantitatively monitor the decomposition rates of superoxide in water. The analysis of the hindered flow kinetics is adequate to examine compounds for SOD activity, and the catalytic activity of the compounds or complexes of the present invention to dismute the superoxide, as shown by the prevented flow analysis, is correlated to treat previous conditions and disease disorders. The total daily dose administered to a host in individual or divided doses may be in amounts, for example, of about 1 to about 100 mg / kg of body weight per day, and more usually about 3 to 30 mg / kg. The unit dose compositions may contain said amounts or submultiples thereof to obtain the daily dose. The amount of active ingredient that can be combined with the carrier materials to produce a single dose form will vary, depending on the treated host and the particular mode of administration. The dose regimen for treating a disease condition with the compounds and / or compositions of this invention is selected in accordance with various 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 used, whether a drug delivery system is used and whether the compound is administered as part of a drug combination. Thus, the dose regime actually used can vary widely and, therefore, may deviate from the preferred dose regimen set forth above. The compounds of the present invention can be administered orally, parenterally, by inhalation spray, rectally or topically in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable excipients, adjuvants and vehicles, as desired. Topical administration may also include the use of transdermal administration, such as transdermal patches or devices for iontophoresis. The term parenteral, as used in the present invention, includes subcutaneous, intravenous, intramuscular, intrasternal, or infusion techniques. Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be formulated according to the known art using suitable wetting or dispersing agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the vehicles and acceptable solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as a solvent or suspension medium. For this purpose, any oil fixed in admixture including synthetic mono- or diglycerides can be used. In addition, fatty acids such as oleic acid are used to prepare injectable products. Suppositories can be prepared for rectal administration of the drug by mixing it with a suitable non-irritating excipient such as cocoa butter and polyethylene glycols which are solid at ordinary temperatures, but liquid at the rectal temperature and which will therefore melt at the rectal temperature. straight and they will 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 can be mixed with at least one inert diluent such as sucrose, lactose or starch. Said dosage forms may also comprise, as in normal practice, other substances than inert diluents, for example, lubricating agents such as magnesium stearate. In the case of capsules, tablets and pills, the dosage forms may also comprise pH regulating agents. Tablets and pills with enteric shells can be further prepared. Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing the inert diluents commonly used in the art, such as water. Said compositions may also comprise adjuvants, such as wetting, emulsifying and suspending agents, and sweetening, flavoring and perfuming agents. Although 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 that are known to be effective against the specific disease state being treated. The compounds or complexes of the invention can also be used as MRI contrast agents. A discussion on the use of contrast agents in MRI can be found in patent application serial No. 08/397469 The contemplated equivalents of the general formulas given above for the compounds and derivatives, as well as the intermediates, are compounds that otherwise correspond to them and have the same general properties such as tautomers of the compounds and such as wherein one or more than the various R groups are simple variations of the substituents as defined in the present invention, for example, wherein R is a higher alkyl group than that indicated, or where the tosyl groups are other nitrogen or oxygen protecting groups, or wherein 0-tosyl is a halide. Anions having a charge other than 1, for example, carbonate, phosphate and hydrogen phosphate, can be used in place of anions having a loading of 1, as long as they do not adversely affect the overall activity of the complex. However, the use of anions that have a charge different from 1 will result in a slight modification of the general formula for the complex described above. Further, where a substituent is designated as, or may be, a hydrogen, the exact chemical nature of a substituent that is not hydrogen at that position, for example, a hydrocarbyl radical or a halogen, hydroxy, amino and similar functional group, does not it is critical as long as it does not adversely affect the general activity and / or the synthesis procedure. In addition, it is contemplated that the manganese (III) complexes will be equivalent to the present manganese (II) complexes. The chemical reactions described above are generally described in terms of their broader 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 scope described. The compounds for which this occurs will be readily recognized by those skilled in the art. In such cases, the reactions can be successfully carried out by conventional modifications known to those skilled in the art, for example, by adequate protection of the interference groups, switching to alternative conventional reagents, by routine modification of the reaction conditions. , and the like, or other reactions described in the present invention or otherwise conventional, will be applicable to the preparation of the corresponding compounds of this invention. In all preparation methods, all starting materials are known, or will be readily prepared from known starting materials.

EXAMPLES All reagents were used as received without purification, unless otherwise indicated. All NMR spectra were obtained on a Varian VXR-300 or VXR-400 nuclear magnetic resonance spectrometer. Qualitative and quantitative mass spectroscopy was carried out on a Finigan MAT90, a Finigan 4500 and a VG40-250T using m-nitrobenzyl alcohol (NBA) or m-nitrobenzyl alcohol / LiCl (NBA-Li). The melting points (p. Of f.) Are uncorrected. The following abbreviations concerning amino acids and their protecting groups are in accordance with the recommendation made by the IUPAC-IUB Commission on Biochemical Nomenclature (Biochemistry 1972, 11, 1726) and common usage. Ala L-Alanine DAla D-Alanine Gly Glycine Se r L-Serine DSe r D-Se rina Bzl Benzyl Boc Te r-butoxy carboni lo Et Ethyl TFA Trifluoroacetic acid DMF Dimethylformamide H0BT »H20? -Hydroxy- (IH) -benzotriazole monohydrate EDO HCl l- (3-Dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride TEA Triethylamine DMSO Dimethyl sulfoxide THF Tetrahydrofuran DPPA Diphenylphosphoryl azide * The abbreviation Cic represents 1,2-cyclohexanediamine (stereochemistry, ie, R, R or S, S, as indicated as such). This makes it possible to use a nomenclature for peptides with a three-letter code in pseudopeptides containing the "residue" of 1,2-cyclohexanediamine.

EXAMPLE 1 A. Synthesis of N- (p-toluenesulfonyl) - (R, R) -l, 2-diaminocyclohexane To a stirred solution of (R, R) -1,2-diaminocyclohexane (300 g, 2.63 moles) in CH2Cl2 (5.00 1) at -10 ° C, a solution of p-toluenesulfonyl chloride (209 g, 1.10 mol) in CH 2 Cl 2 (5.00 1) was added dropwise over a period of 7 hours, maintaining the temperature at -5 to -10. ° C.

The mixture was allowed to warm to room temperature while stirring overnight. The mixture was concentrated in vacuo to a volume of 3 liters, and the white solid was removed by filtration. The solution was then washed with H 2 O (10 x 11), and dried over MgSO 4. Removal of the solvent in vacuo gave 286 g (97.5% yield) of the product as a yellow crystalline solid: iH NMR (CDCl 3) d 0.98-1.27 (m, 4H), 1.54-1.66 (m, 2H), 1.81-1.93 (m, 2H), 2.34 (dt, J = 4.0, 10.7 Hz, 1H), 2.42 (s, 3H), 2.62 (dt, J = 4.2, 9.9 Hz, 1H), 7.29 (d, J = 8.1 Hz, 2H), 7.77 (d, J = 8.3 Hz, 2H); MS (LRFAB-DTT-DTE) m / z 269 CM + H] +.

B. Synthesis of N- (p-toluenesulfonyl) -N '- (Boc) - (RR) -1.2- diaminocyclohexane To a stirred solution of N- (p-toluenesulfonyl) - (R, R) -l, 2-diaminocyclohexane prepared as in Example IA (256 g, 0.955 mol) in THF (1.15 1), a 1 N aqueous NaOH solution (1.15 1, 1.15 mol) was added. Then, di-t-butyldicarbonate (229 g, 1.05 mol) was added, and the resulting mixture was stirred overnight. The layers were separated, and the aqueous layer was adjusted to pH 2 with IN HCl, and saturated with NaCl. The aqueous solution was then extracted with CH2Cl2 (2 x 500 ml), and the extracts and the THF layer were combined and dried over MgSO-4. The solvent was removed in vacuo to give a yellow solid. The crude product was purified by crystallization from a THF-ether-hexanes mixture to give 310 g (88.1% yield) of the product as a white crystalline solid: p. of f. of 137-139 ° C; NMR * H (CDCl 3) d 1.04-1.28 (m, 4H), 1.44 (s, 9H), 1.61-1.69 (m, 2H), 1.94-2.01 (m, 2H), 2.43 (s, 3H), 2.86 ( brs, 1H), 3.30 (br d, J = 9.6 Hz, 1H), 4.37 (br d, J = 6.7 Hz, 1H), 5.48 (br d, J = 4.6 Hz, 1H), 7.27 (d, J = 9.7 Hz, 2H), 7.73 (d, J = 8.1 Hz, 2H); MS (LRFAB, NBA-Li) m / z 375 [M + Li] +.

C. Synthesis of Boc- (RR) -CIc (Ts) -gly-OMe To a stirred solution of N- (p-toluenesulfonyl) -N '- (Boc) - (R, R) -l, 2-diaminocyclohexane prepared as in Example IB (310 g, 0.841 mol) in anhydrous DMF (3.11 1) at 0 * C, NaH (37.4 g- 60% in oil, 0.934 mol) was added in portions, and the resulting mixture was stirred for 30 minutes. minutes Then, methyl bromoacetate (142 g, 0.925 mol) was added dropwise over 45 minutes, and the mixture was allowed to warm to room temperature while stirring overnight. After stirring for 17 hours, the solvent was removed in vacuo, and the residue was dissolved in ethyl acetate (3 1) and H 2 O (1 1). The ethyl acetate solution was washed with saturated NaHCO 3 (11), saturated NaCl (500 L), and dried over MgSO 4.; . The solvent was removed in vacuo and the resulting oil was dissolved in ether. Crystallization by the addition of hexanes gave 364 g (98% yield) of the product (TLC (98: 2 CHCl3-MeOH / silica gel / UV), and showed that the product contained about 5% starting material) as colorless needles: boiling point of pure mixture 151-2 ° C; 1H-NMR (CDCl3) d 1.11-1.22 (m, 4H), 1.45 (s, 9H), 1.64-1.70 (m, 3H), 2.16-2.19 (m, 1H), 2.43 (s, 3H), 3.34-3.40 (m, 2H), 3.68 (s, 3H), 4.06 (ABq, J = 18.5 Hz, "= 155 Hz, 2H), 4.77 (br s 1H), 7.30 (d, J = 8.3 Hz, 2H), 7.82 (d, J = 8.3 Hz, 2H); MS (LRFAB, DTT-DTE) m / z 441 [M + H] +.

D. Synthesis of Boc- (RR) -Cyc (Ts) -Gly-OH To a stirred solution of impure Boc- (R, R) -Cic (Ts) -Gly-0Me prepared as in Example 1C (217 g, 0.492 moles) in MeOH (1.05 1), a 2.5N aqueous NaOH solution (295 L, 0.737 moles) was slowly added, and the resulting solution was stirred for 2 hours. The solvent was removed in vacuo and the residue was dissolved in H2O (1.5 1). The solution was filtered to remove a small amount of solid, and washed with ether (7 1 1) to remove the impurity (compound IB), which after drying the combined washes over MgSO 4 and to remove the solvent in vacuo, allowed recover 8.37 g. The pH of the aqueous solution was then adjusted to 2 with IN HCl, and the product was extracted with ethyl acetate (3 x 11). The extracts were combined, washed with saturated NaCl (500 mL), and dried over MgSO 4. The solvent was removed in vacuo, and the residual ethyl acetate was removed by coevaporation with ether (500 mL) and then CH2CI2 (500 mL) to give 205 g (97.6% yield) of the product as a white foam: NMR * H (CDCl 3) d 1.15-1.22 (m, 4H), 1.48 (s, 9H), 1.55 NMR-1.68 (m, 3H), 2.12-2.15 (m, 1H), 2.43 (s, 3H), 3.41-3.49 ( m, 2H), 3.97 (ABq, J = 17.9 Hz, "= 69.6 Hz, 2H), 4.79 (br s, 1H), 7.31 (d, J = 8.1 Hz, 2 H), 7.77 (d, J = 8.3 Hz, 2H), 8.81 (br s, 1 H); MS (LRFAB, NBA-Li) m / z 433 [M + Li] +.

E. Synthesis of Boc- (RR) -Cyc (Ts) -Gly-Gly-OEt A Boc- (R, R) -Cyc (Ts) -Gly-OH (18.1 g, 43.1 mmol) in DMF (480 mL) H0Bt »H20 (7.92 g, 51.7 mmol) and EDC» HC1 (9.91 g, 51.7 mmol) were added, and the resulting mixture was allowed to stir for 20 minutes at room temperature. To this solution was added Gly0Et »HCl (6.0 g, 43.1 mmol) and TEA (7.2 mL, 51.7 mmol), and the resulting mixture was allowed to stir for 16 hours. The DMF was evaporated, and the residue was partitioned between water (250 mL) and EtOAc (400 mL). The EtOAc layer was separated and washed with IN KHSO4 (250 L), water (250 mL), saturated NaHCO3 (250 L) and brine (250 L), and dried (NaSO4 - The filtration steps and concentration produced 21.9 g (99% yield) of pure product as a white foam: H-NMR (DMSO-de) d 1.00-1.10 (m, 1H), 1.19 (t, J = 7.6 Hz, 3H), 1.38 (s, 9H), 1.50-1.56 (m, 3H), 1.75-1.84 (m, 1H), 2.38 (s, 3H), 3.30-3.40 (bs, 2H), 3.75-4.01 (complex m, 4H), 4.08 (q , J = 7.6 Hz, 2H), 6.05 (bs, 1H), 7.32 (d, J = 8.0 Hz, 2H), 7.77 (d, J = 8.0 Hz, 2H), 8.32 (t, J = 7.2 Hz, 1H ); MS (HRFAB) m / z 518.2551 (M + Li) +; 518.2512 calculated for C24 H37N3O7SLÍ.

F. Synthesis of cyc (Ts) salt -Glv-Gly-OEt TFA To a solution of Boc-Cic (Ts) -Gly-Gly-OEt (21.2 g, 41.4 mmoles) in CH2Cl2 (180 mL) was added TFA ( 44 mL), and the resulting mixture was stirred at room temperature for 30 minutes. The solution was concentrated, and the residue was dissolved in ether (50 L), and precipitated with hexanes (500 L). The solvents were decanted, and the residue was washed with 10: 1 hexanes / ether (500 mL). The final residue was dried completely under high vacuum to yield 20.7 g (95% yield) of the product as a tan foam: NMR * H (DMSO-dβ) d 0.85-0.96 (m, 1H), 1.03-1.31 ( complex m, 7H), 1.09 (t, J = 7.6 Hz, 3H), 2.00 (m, 1H), 2.39 (s, 3H), 3.02 (bs, 1H), 3.62 (, 1H), 3.82-4.05 (m , 4H), 4.10 (q, J = 7.6, 2H), 7.41 (d, J = 8.0 Hz, 2H), 7.67 (d, J = 8.0 Hz, 2H), 8.25 (bs, 3H), 9.09 (t, J s 5.63 Hz, 1H). MS (HRFAB) m / z 418.1990 (M-TFA + Li) +; 418.1988 calculated for O H29 3OSS.

G. Synthesis of Boc-Orn (Z) -Cyc (Ts) -Gly-Gly-OEt To Boc-0rn (Z) -0H (8.37 g, 22.8 mmol) in DMF (200 mL) was added H0Bt «H20 (4.29 g, 27.4 mmoles) and ED HC1 (5.25 g, 27.4 mmoles), and the resulting solution was stirred for 20 minutes at room temperature. To this solution was added the salt of Cic (Ts) -Gly-Gly-OEt TFA (12.0 g, 22.88 mmol) and TEA (3.82 mL, 27.4 mmol), and the stirring was then maintained for 16 hours. The DMF was evaporated, and the residue was partitioned between water (200 mL) and EtOAc (250 mL). The ETOAc layer was separated and washed with IN KHSOA (150 mL), water (150 mL), saturated NaHCO3 (150 mL) and brine (150 mL), and dried (MgSO-Filtration and concentration steps yielded 15.1). g (87% yield) of the product as a white foam: H-NMR (DMS0-de) d 1.00-1.94 (complex m, 12 H), 1.15 (t, J = 7.4 Hz, 3H), 2.38 (s) , 3H), 2.98 (bs, 2H), 3.30-3.46 (m, 2H), 3.70-3.82 (m, 4H), 3.90-4.02 (m, 1H), 4.05 (t, J = 7.4 Hz, 2H), 5.00 (s, 2H), 6.43 (m, 1H), 7.17 (m, 1H), 7.20-7.37 (m, 8H), 7.78 (m, 2H), 8.30 (bs, 1H); MS (LRFAB, NBA + HCl) m / z 760 (M + H) + H. Synthesis of Orn (Z) -Cyc (Ts) -Gly-Gly-QEt salt TFA To a solution of Boc-0rn (Z) -Cyc (Ts) -Gly-Gly-0Et (14.5 g, 19.1 mmol) in CH2Cl2 (120 mL) was added TFA (30 mL), and the resulting solution was stirred at room temperature for 30 minutes. The solution was evaporated, and the residue was triturated with ether (100 L). The ether was decanted, and the residue dried completely under high vacuum to yield 15.5 g (> 100% yield, TFA-containing) of the product as an orange foam: * H NMR (DMSO-dβ) d 0.97-1.93 (m complex, 12H), 1.16 (t, J = 7.4 Hz, 3H), 2.38 (s, 3H), 2.98 (bs, 2H), 3.31-3.50 (m, 2H), 3.71-3.91 (m, 4H), 3.97 -4.04 (m, 1H), 4.08 (q, J - 7.4 Hz, 2H), 5.00 (s, 2H), 7.23-7.39 (m, 8H), 7.77-7.81 (m, 2H), 8.18 (bs, 3H) ), 8.41 (bs, 1H); MS (LRFAB, NBA + HCl) m / z 660 (M-TFA +.

I. Synthesis of Boc-Glv-Orn (Z) -Cyc (Ts) -Gly-Gly-QEt To a solution of Boc-Gly-OH (3.36 g, 19.2 mmol) in DMF (220 mL) was added H0Bt »H20 (3.52 g, 23.0 mmol) and EDC »HC1 (4.41 g, 23.0 mmol), and the resulting solution was stirred for 20 minutes at room temperature. To this solution was added salt of 0rn (Z) -Cyc (Ts) -Gly-Gly-0Et TFA (14.8 g, 19.2 mmol) and TEA (3.20 mL, 23.0 mmol), and the stirring was then maintained for 12 hours. The DMF was evaporated, and the residue was partitioned between water (200 mL) and EtOAc (350 mL). The layers were separated, and the EtOAc layer was washed with KHS0¿ IN (150 L), water (150 mL), saturated NaHCO3 (150 mL) and brine (150 mL), and dried (MgSO *). The filtration and concentration steps yielded 13.7 g (87% yield) of the product as a white foam: RMNaH (DMSO-dβ) d 0.96-1.10 (m, 2H), 1.17 (t, J = 7.4 Hz, 3H), 1.38 (s, 9H), 1.35 - 2.00 (complex m, 10H), 2.97 (m, 2H), 3.60 (bs, 2H), 3.67 - 3.84 (m, 4H), 3.93 - 4.03 (m, 3H), 4.06 (q, J = 7.4 Hz, 2H), 6.92 (bs, 1H), 7.19 (m, 1H), 7.24 - 7.37 (, 7H), 7.60 (d, J = 8.3 Hz, 1 H), 7.76 (m, 2H), 7.38 (bs, 1H). MS (LRFAB, NBA + Li) * m / z 823 (M + Li) +.

J. Synthesis of Boc-Gly-0rn (Z) -Cyc (Ts) -Gly-Gly-0H To a solution of Boc-Gly-0rn (Z) -Cic (Ts) -Gly-Gly-0Et (13.3 g, 16.3 mmol) in methanol (100 mL) was added 1 N NaOH (25 mL). The resulting mixture was stirred at room temperature and monitored by TLC. After 2 hours, the reaction concluded. The methanol was evaporated and water (50 L) was added to the residue. This aqueous phase was washed with EtOAc (2 x 100 mL), and the EtOAc layers were discarded. The pH was reduced to 3.5 with 1 N KHSO 4, and the aqueous phase was extracted with EtOAc (3 x 100 mL). The combined EtOAc layers were dried (MgSO, filtered and concentrated to yield 11.7 g (91% yield) of the product as a white foam: H-NMR (CDCl3) d 0.98-1.25 (m, 2H), 1.38 (s) , 9H), 1.40 - 1.92 (m, 10H), 2.38 (s, 3H), 2.97 (m, 2H), 3.62 (bs, 2H), 3.75 - 3.85 (m, 3H), 3.95 - 4.05 (m, 2H) ), 5.01 (s, 2H), 6.96 (bs, 1H), 7.28 (m, 1H), 7.25 - 7.38 (m, 7H), 7.61 (d, J = 8.4 Hz, 1H), 7.78 (m, 2H) , 8.25 (bs, 1H).

K. Synthesis of Gly-Orn (Z) -Cyc (Ts) -Gly-Gly-OH salt TFA To a solution of Boc-Gly-0rn (Z) -Cyc (Ts) -Gly-Gly-0H (11.2 g , 14.3 mmoles) in CH2Cl2 (100 mL) was added TFA (24 mL), and the resulting solution was stirred for 30 minutes at room temperature. The solution was concentrated and triturated with ethyl ether (500 mL). The filtration step produced 11.3 g (99% yield) of the product as a powder, white: H-NMR1 (DMSO-ds) d 0.95-1.98 (complex m, 12H), 2.39 (s, 3H), 3.01 (m, 2H), 3.38 (m, 1H), 3.65 - 4.10 (complex m, 7H), 4.18 (q, J = 7.4 Hz, 1H), 5.02 (s, 2H), 7.24 -7.40 (m, 9H), 7.77 - 7.85 (m, 2H), 8.13 (bs, 3H), 8.31 (bs, 1H), 8.42 (d, J = 8.3 Hz, 1 HOUR); MS (HRFAB) 689.2953 (M-TFA) +; 689.2969 calculated for C32H45N6O9S.

L. Synthesis of Cyclo- (Gly-Qrn (Z) -Cic (Ts) -Gly-Gly-) A solution of salt of Gly-Orn (Z) -Cic (Ts) -Gly-Gly-OH TFA (5.0 g , 6.23 mmoles) in degassed dry DMF (1520 mL) was treated with TEA (1.74 mL, 12.5 mmol), and cooled to -40 * C. DPPA (1.64 mL, 7.60 mmol) was added dropwise over 10 minutes, and the reaction was then stirred at -40 ° C for 3 hours. After this time, the reaction was placed in a -2 ° C bath, and then allowed to stand at this temperature for 16 hours. Water (1520 mL) was added, and the resulting solution was stirred with mixed bed ion exchange resin (750 g) for 6 hours at room temperature. The resin was filtered, and the solution was concentrated to a volume of ~ 100 mL (DMF). The addition of ethyl ether (500 mL) produced a solid residue which was redissolved in methanol (100 mL), and was again precipitated by the addition of ethyl ether (500 mL). Filtration produced 3.26 g (78% yield) of the product as a white powder: 1 H-NMR (CDCl 3) d 0.96-2.10 (complex m, 14H), 2.37 (bs, 3H), 2.68-3.05 (m, 3H), 3.42 - 3.90 (complex m, 8H), 4.14 (m, 1H), 4.20 (m, 1H), 4.97 - 5.08 (m, 3H), 6.42 (d, J -8.4 Hz, 1H), 7.19 (d, J = 8.0 Hz, 1H), 7.20 - 7.39 (m, 7H), 7.65 - 7.78 (m, 2H), 9.15 (bs, 1H), 9.22 (bs, 1H); MS (HRFAB) m / z 671.2842 (M + H) +; 671.2863 calculated for C32HA3Ne08S.

M. Synthesis of cyclo-Gly-Orn-Cic (Ts) -Gly-Gly-) To a solution of cyclo- (Gly-0rn (Z) -Cic (Ts) -Gly-Gly-) (3.94 g, 5.90 mmol) in methanol (40 mL) was added Pd (black) (1.0 g) and ammonium formate (2.0 g). The reaction was refluxed for 2 hours and allowed to cool. The mixture was filtered under argon through a pad of celite, and the filtrate was concentrated to yield 2.86 g (89% yield) of the product as a white foam: RMNiH (DMSO-dβ) d 0.94-2.22 (complex m, 12H), 2.39 (s, 3H), 2.55 - 2.95 (m, 7H), 3.42 - 3.89 (complex, 9H), 4.11 (m, 1H), 4.39 (m, 1H), 6.43 (d, J = 8.4 Hz , 1H), 7.27 (d, J = 9.3 Hz, 1H), 7.25-7.45 (m, 2H), 7.64-7.80 (m, 2H), 9.12-2.29 (m, 2H); MS (HRFAB) m / z 537.2511 (M + H) +; 537.2495 calculated for C2¿H36N6S0e.

N. Synthesis of cyclo- (Gly-Qrn (Litocolil) -Cic (Ts) -Gly-Gly-) To a solution of cyclo- (Gly-0rn-Cic (Ts) -Gly-Gly-) (1.0 g, 1.9 mmoles) in CHCl3 (25 mL) was added active NHS ester of lithocholic acid (881 mg, 1.9 mmol), and the resulting mixture was then stirred for 16 hours. The addition of ethyl ether (50 mL) yielded a solid. Filtration yielded 946 mg (56% yield) of the product as a tan powder: RM? H (CD3OD) d 0.66 (m, 3H), 0.93 (bs, 6H), 0.94 -2.37 (complex m, 48H) , 2.43 (s, 3H), 2.80 - 4.60 (b, 14H), 7. 39 (bs, 2H), 7.80 (bs, 2H); MS (HRFAB) m / z 895.5432 (M + H) +; 895.5367 calculated for C48H75N6O8S. 0. Synthesis of 2.3- (R.R) -cyclohexane-6- (S) -. { 3- Clcycolylamino) propyl} -l.4.7.10.13-penta-azaciclopentadecane To a suspension of cyclo- (Gly-0rn (Litocolil) -Cic (Ts) -Gly-Gly-) (2.70 g, 3.00 mmol) in THF (50 L) was added hydride of lithium-aluminum (51.0 mL of a 1.0 M solution). The resulting mixture was then refluxed for 16 hours. The reaction mixture was cooled to -20 ° C and warmed (carefully) with 5% Na S04 (30 L), followed by methanol (30 mL). This solution was stirred at room temperature for 1 hour and concentrated to a dry powder. The powder was triturated with ethyl ether (3 x 200 mL) and filtered. The ether was concentrated, and the oil was recrystallized from acetonitrile to yield 800 mg (40% yield) of the product as a colorless oil: NMR? H (CeDe) d 0.64 (s, 3H), 0.67 (s, 3H ), 0.88 (d, J = 3.0 Hz, 3H), 0.84-2.61 (complex m, 52H), 2.38-2.95 (complex m, 14H), 3.49 (m, 3H); 13 C NMR (CDCl 3) d 71.4, 63. 1, 62.6, 61.8, 58.2, 56.5, 56.1, 51.5, 50.4, 50.1, 48.3, 47.9, 46.1, 45.7, 42.6, 42.1, 40.4, 40.1, 36.4, 35.8, 35.7, . 6, 35.4, 34.5, 31.9, 31.7, 31.6, 30.8, 30.5, 29.4, 28.3, 27. 2, 26.4, 26.2, 24.9, 24.2, 23.4, 20.8, 18.6, 12.0; MS (LRFAB, NBA + Li) m / z 677 (M + Li) +.

P. Synthesis of [dichloro 2.3- (RR) -cyclohexane-6- (S) - (3- (litho-colylamino) -propyl.} -1 .7.10.13-penta-azaciclopentadecane of manganese CID 3 2,3- (R, R) -cyclohexane-6- (S) -. {3- (litho-colylamino) propyl} -1,4,7,10,13-penta-azacyclopentadecane prepared as in example 10 (547 mg, 0.817 mmoles), was added to a hot solution of anhydrous methanol (50 mL) containing manganese (II) chloride (103 mg, 0.818 mmol) under a dry nitrogen atmosphere, after refluxing for 2 hours, the solution The residue was dissolved in a solvent mixture of THF (35 mL) and ethyl ether (5 L) and filtered through a pad of celite.The concentration and trituration with ethyl ether produced after filtration. 512 mg (79% yield) of the complex as a white solid: FAB mass spectrum (NBA) m / z 760 [M-C13 +, analysis calculated for C41H78N60MnC12: C, 61.79; H, 9.87; N, 10.55; Cl 8.90 Found: C, 62.67; H, 9.84; N, 8.04; Cl, 8.29.

EXAMPLE 2 Flow impeded kinetics analysis The impeded flow kinetics analysis has been used to determine whether a compound can catalyze the dismutation of superoxide (Riley, D.P., Rivers, W.J. and Weiss, R.H., "Stopped-Flo Kinetic Analysis for Monitoring Superoxide Decay in Aqueous Systems," Anal. Biochem, 196, 344-349 [1991].

To obtain consistent and accurate measurements, all reagents were biologically clean and free of metal. To achieve this, all pH regulators (Calbiochem) were pH regulators free of metal and biological grade, and were handled with utensils that had been washed first with 0.1 N HCl, followed by purified water, followed by a rinse in a bath of EDTA at 10 * M at pH 8, and followed by a rinse with purified water and drying at 65 ° C for several hours. Dry DMSO solutions of potassium superoxide (Aldrich) were prepared under a dry inert argon atmosphere in a Vacuum Atmospheres dry glove compartment using dry glassware. The DMSO solutions were prepared immediately before each impeded flow experiment. A mortar and pestle were used to crush the yellow solid of potassium superoxide (~ 100 mg). The powder was then ground with a few drops of DMSO, and the suspension was transferred to a flask containing another 25 mL of DMSO. The resulting suspension was stirred for half an hour, and then filtered. This procedure reproducibly gave concentrations of ~2 mM superoxide in DMSO. These solutions were transferred to a glove bag under nitrogen in sealed jars before loading the syringe under nitrogen. It should be noted that DMSO / superoxide solutions are extremely sensitive to water, heat, air and foreign metals. A pure and fresh solution has a very faint yellowish tint. Water was supplied for pH buffer solutions from a domestic deionized water system to a Barnstead Nanopure Ultrapure 550 series fluvial system, and then distilled twice, first from alkaline potassium permanganate, and then from a dilute solution of EDTA. For example, a solution containing 1.0 g of potassium permanganate, 2 liters of water and additional sodium hydroxide necessary to bring the pH to 9.0 was added to a 2 liter flask adapted with a solvent distillation head. This distillation will oxidize any trace of organic compounds in the water. The final distillation was carried out under nitrogen in a 2.5 liter flask containing 1500 mL of water from the first distillation and EDTA at 1.0 x 106M. This step will remove trace metals remaining from ultrapure water. To prevent the EDTA mist from volatilizing on the reflux arm towards the distillation head, the 40 cm vertical arm was packed with glass spheres and wrapped with insulation. This system produces deoxygenated water that can be measured to have a conductivity of less than 2.0 nanomhos / cm2. The impeded flow spectrometer system was designed and manufactured by Kinetic Instruments Inc. (Ann Arbor, MI), and was interfaced with a MAC IICX personal computer. The software for the prevented flow analysis was provided by Kinetics Instrument Inc., and was registered in QuickBasic controllers with MacAdios. The typical volumes of the injector (0.10 mL of pH regulator and 0.006 mL of DMSO) were calibrated, so that a large excess of water over the DMSO solution was mixed therewith. The actual ratio was about 19/1, so that the initial concentration of the superoxide in the aqueous solution was on the scale of 60 to 120 μM. Because the published extinction coefficient for superoxide in H2O at 245 nm is ~ 2250 M-1 cm-1 (1), an initial absorbance value of approximately 0.3 to 0.5 would be expected for a path length cell of 2 cm, and this was observed experimentally. Aqueous solutions were prepared which are then mixed with the superoxide DMSO solution using concentrations of 80 mM Hepes pH buffer, pH 8.1 (free acid plus Na form). One of the syringes in the reservoir was filled with 5 ml of the DMSO solution, while the other was filled with 5 ml of the aqueous buffer solution. The complete injection block, the mixer and the spectrometer cell were submerged in a circulating water bath with a thermostat with a temperature of 21.0 ± 0.5 ° C. Before starting the data collection for a decomposition of the superoxide, an average of the baseline was obtained by injecting several bursts of the pH regulator and DMSO solutions into the mixing chamber. These bursts were averaged and stored as the baseline. The first bursts to be collected during a series of runs were with aqueous solutions that did not contain a catalyst. This ensured that each series of tests were free of contamination capable of generating first-order superoxide decomposition profiles. If the decompositions observed for several bursts of the pH regulator solution were of second order, solutions of manganese (II) complexes could be used. In general, the potential catalyst for SOD was examined over a wide range of concentrations. Since the initial concentration of the superoxide after mixing the DMSO with the aqueous pH regulator was ~ 1.2 x 10"*, we wanted to use a concentration of the manganese (II) complex that was at least 20 times lower than that of the substrate superoxide Accordingly, compounds for SOD activity were generally examined using concentrations ranging from 5 x 10 ~ 7 to 8 x 10 ~ 6 M. The data obtained from the experiment were entered into a suitable mathematical program (for example, Cricket Graph ), so that standard analyzes could be performed on kinetic data The catalytic rate constant for the dismutation of the superoxide by the manganese (II) complex of example 1 was determined from the linear plot of the observed rate constants ( K-obe) against the concentration of manganese (II) complexes K-0bs values were obtained from the linear graphs of the absorbance ln at 245 nm against time for the dismutation of superoxide by the manganese (II) complex. It was determined that the Kcat (M_1 sec-1) of the manganese complex (II) of Example 1 at pH: 8.1 and 21 ° C is 0.77 x 10 + * M- * sec- i. The manganese (II) complex of the nitrogen-containing macrocyclic ligand in Example 1 is an effective catalyst for the dismutation of superoxide, as can be seen from the previous Kcat.

Claims (17)

NOVELTY OF THE INVENTION CLAIMS

1. - A compound that is a complex represented by the formula: wherein R, R ', R, R'i, R2, R'2, R3, R'3, R <; , R 'A, Rs, Rs, Re, R'ß, R7, R'7 RSs, R'ß, R9 and R'9 independently represent alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, alkylcycloalkyl , alkenylcycloalkyl, alkylcycloalkenyl, alkenylcycloalkenyl, heterocyclic aryl and aralkyl radicals and radicals attached to carbon a of the oc-amino acids; or Ri or R'i and. or R'2, R3 or R'3 and R < or R'4, Rs or R'5 and Rs or R'6, R? or R'7 and Rβ or R'ß and R9 or R'9 and R or R 'together with the carbon atoms to which they are attached independently form a saturated, partially saturated or unsaturated cycle having from 3 to 20 carbon atoms. carbon; or R or R 'and Ri or R'i, R2 or R'2 and R3 or R'3, ¿. or R '¿> and Rs or R's, Re or R'ß and R7 or R'7 and Rβ or R'ß and R or R'9 together with the carbon atoms to which they are attached independently form a heterocycle having from 2 to 20 atoms of carbon, provided that when the nitrogen-containing heterocycle is an aromatic heterocycle that does not contain a hydrogen bound to nitrogen, hydrogen bound to nitrogen is absent in said formula, whose nitrogen is also in the macrocycle and the R groups attached thereto carbon atoms of the macrocycle; and combinations thereof; characterized in that (1) one to five of the "R" groups are bound to biomolecules by a linker group, (2) one of X, Y and Z is attached to a biomolecule by a linker group or (3) one to five of the "R" groups and one of X, Y and Z are joined to biomolecules by a linker group; and said biomolecules are independently selected from the group consisting of steroids, carbohydrates, fatty acids, amino acids, peptides, proteins, antibodies, vitamins, lipids, phospholipids, phosphates, phosphonates, nucleic acids, enzyme substrates, enzyme inhibitors and receptor substrates. of enzymes, and said linker group is derived from a substituent attached to said "R" group or said X, y and Z which reacts with the biomolecule and is selected from the group consisting of -NH2, -NHR10, -SH, - OH, -C00H, -COOR or, -CONH2, -NCO, -NCS, -C00X ", alkenyl, alkynyl, halide, tosylate, mesylate, tresylate, triflate and phenol, wherein Rio is alkyl, aryl or alkaryl and X" it is a halide; and wherein X, Y and Z are ligands independently selected from the group consisting of halide, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hidroxo, alquilperoxo, arilperoxo, ammonia, alkylamino, arylamino, amino heterocycloalkyl, amino heterocicloaril amine oxides, hydrazine, hydrazine alkyl, aryl hydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkyl nitrile, aryl nitrile, alkyl isonitrile, aryl isonitrile, nitrate, nitrite, azido, alkyl sulfonic acid, arylsulfonic acid, alkyl sulfoxide, dimethyl aryl, alkyl aryl sulfoxide, alkyl acid sulfenic, aryl acid sulfenic, alkyl sulphinic acid, aryl sulphinic acid, alkyl acid thiol carboxylic acid, aryl acid thiol carboxylic acid, alkyl acid thiol thiocarboxylic, aryl acid thiol thiocarboxylic acid alkyl ester, aryl carboxylic 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 oxide, alkyl phosphine, phosphine oxide aryl oxide, alkyl aryl phosphine sulfide, alkyl phosphine, phosphine sulfide aryl sulfide alkyl aryl phosphine acid alkyl phosphonic acid, aryl phosphonic acid, alkyl phosphinic acid, aryl phosphinic acid, alkyl acid phosphinous aryl acid phosphinous, phosphate, thiophosphate, phosphite, pirofosfita, triphosphate, hydrogen phosphate, dihydrogen phosphate, guanidino alkyl, aryl guanidino, alkyl aryl guanidino, alkyl carbamate, aryl carbamate, alkyl aryl carbamate, alkyl thiocarbamate, aryl thiocarbamate, alkyl thiocarbamate, alkyl dithiocarbamate, aryl dithiocarbamate, alkyl aryl dithiocarbamate, bicarbonate, carbonate, perchlorate, chlorate, chlorite, hypochlorite, perbromate, bromate, bromite, hypobromite, tetrahalogenomanate, tefluoroborate, hexafluoroantimonate, hypophosphite, iodate, periodate, metaborate, tetraaryl borate, tetraalkyl borate, tartrate, salicylate, succinate, citrate, ascorbate, saccharinate, amino acid, hydroxamic acid, thiotosylate, and ion exchange resin anions, or the corresponding anions thereof, or X, Y and Z independently bind to one or more of the groups "R", and n is 0 or 1.

2. The compound of claim 1, characterized in that 1 to 2 of the "R" groups are bound to biomolecules by a linker group, and none of X, Y and Z is linked to a biomolecule by a linker group.

3. The compound of claim 1, characterized in that one of X, Y and Z is attached to a biomolecule by a linker group, and none of the "R" groups is bound to biomolecules by a linker group.

4. The compound of claim 1, characterized in that a maximum of a group "R" attached to the carbon atoms of the macrocycle located between nitrogen atoms has a biomolecule linked by a linker group.

5. The compound of claim 1, characterized in that at least one of the "R" groups, in addition to the "R" groups that are bound to biomolecules by a linker group, are independently selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, alkaryl, aryl, heterocycles and radicals attached to the α-a-amino acid carbon, and the remaining "R" groups are independently selected from hydrogen, saturated, partially saturated or unsaturated cycles or a nitrogen-containing heterocycle .

6. The compound of claim 5, characterized in that at least two of the "R" groups, in addition to the "R" groups that are bound to biomolecules by a linker group, are independently selected from the group consisting of alkyl, cycloalkyl, cycloalkylalkyl, aralkyl, alkaryl, aryl, heterocycles and radicals attached to the carbon oc of a-amino acids.

7. The compound of claim 5, characterized in that at least one of the "R" groups, in addition to the "R" groups that are bound to biomolecules by a linker group, are alkyl, and the remaining "R" groups they are independently selected from hydrogen or saturated, partially saturated or unsaturated cycles.

8. The compound of claim 1, characterized in that at least one of Ri or R'i and R2 or R'2, R3 or R'3 and R <; or R'4, Rs or R's and Rß or R'ß, R7 or R'7 and Rβ or R's R9 or R'9 and R or R 'together with the carbon atoms to which they are attached represent a saturated cycle, partially saturated or unsaturated having from 3 to 20 carbon atoms, and the remaining "R" groups in addition to the "R" groups that are bound to biomolecules by linker groups, are independently selected from hydrogen, nitrogen containing heterocycles or alkyl groups.

9. The compound of claim 8, characterized in that at least two of Ri or R'i and R2 or R'2, R3 or R'3 and "or R'4, Rs or R's and Rß or R'ß , R7 or R'7 and Rβ or R'ß and R9 or R'9 and R or R 'together with the carbon atoms to which they are attached represent a saturated, partially saturated or unsaturated cycle having from 3 to 20 atoms of carbon, and the remaining "R" groups in addition to the "R" groups that are bound to biomolecules by linker groups, are independently selected from hydrogen, nitrogen containing heterocycles or alkyl groups.

10. The compound of claim 8, characterized in that said saturated, partially saturated or unsaturated cycle is cyclohexyl.

11. The compound of claim 10, characterized in that said remaining "R" groups, in addition to the "R" groups that are bound to biomolecules by linker groups, are independently selected from hydrogen or alkyl groups.

12. - The compound of claim 1, characterized in that said R or R 'and Ri or R'i, R2 or R'2 and R3 or R'3, or R'4 and Rs or R's, Rß or R'ß and R7 or R'7 and Rβ or R'ß and R9 or R'9 together with the carbon atoms to which they are attached are found to form a nitrogen-containing heterocycle having 2 to 20 carbon atoms, and the remaining "R" groups in addition to the "R" groups that are bound to biomolecules by a linker group are independently selected from hydrogen, saturated, partially saturated or unsaturated cycles or alkyl groups.

13. The compound of claim 1, characterized in that X, Y and Z are independently selected from the group consisting of halide, organic acid, nitrate and bicarbonate anions.

14. the pharmaceutical composition in unit dosage form useful for dismutating the superoxide comprising (a) a therapeutically or prophylactically effective amount of a complex of claim 1 and (b) a pharmaceutically acceptable non-toxic excipient, adjuvant or vehicle.

15. The use of a complex of claim 1 for preparing a medicament for preventing or treating a disease or disorder that is mediated, at least in part, by superoxide.

16. The use according to claim 15, characterized in that said disease or disorder is selected from the group consisting of ischemic reperfusion injury, surgically induced ischemia, inflammatory bowel disease, rheumatoid arthritis, ate rosary, thrombosis, platelet aggregation, injury and tissue damage induced by oxidants, osteoarthritis, psoriasis, organ transplant rejection, radiation induced injury, fulminant crisis, acute pancreatitis, insulin dependent diabetes mellitus, respiratory distress of the child and adult, metastasis and carcinogenesis.

17. The use according to claim 16, characterized in that said disease or disorder is selected from the group consisting of ischemic reperfusion injury, fulminating crisis, ate roseros and inflammatory bowel disease.