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WO1996039409A1 - Oxydants et reducteurs nitrosyles et nitres de superoxydes - Google Patents

Oxydants et reducteurs nitrosyles et nitres de superoxydes Download PDF

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Publication number
WO1996039409A1
WO1996039409A1 PCT/US1996/008406 US9608406W WO9639409A1 WO 1996039409 A1 WO1996039409 A1 WO 1996039409A1 US 9608406 W US9608406 W US 9608406W WO 9639409 A1 WO9639409 A1 WO 9639409A1
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Prior art keywords
compound
group
superoxide
formula
ligand
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PCT/US1996/008406
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English (en)
Inventor
Jonathan S. Stamler
James D. Crapo
Irwin Fridovich
Brian J. Day
David S. Garvey
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Nitromed, Inc.
Duke University
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Priority to AU60310/96A priority Critical patent/AU6031096A/en
Publication of WO1996039409A1 publication Critical patent/WO1996039409A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/02—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings
    • C07D263/04—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings not condensed with other rings having no double bonds between ring members or between ring members and non-ring members
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/92—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with a hetero atom directly attached to the ring nitrogen atom
    • C07D211/94—Oxygen atom, e.g. piperidine N-oxide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D259/00—Heterocyclic compounds containing rings having more than four nitrogen atoms as the only ring hetero atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/022—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -X-C(=O)-(C)n-N-C-C(=O)-Y-; X and Y being heteroatoms; n being 1 or 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/0606—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

  • the invention relates to novel nitroso and nitro derivatives of compounds which oxidize and/or reduce superoxide either catalytically or stoichiometrically and to pharmaceutical compositions comprising the nitroso or nitro derivative of the invention together with a pharmaceutically acceptable carrier.
  • the invention also relates to their use in treating a variety of disorders.
  • mutagenic alterations in DNA Possibly the most important source of mutagenic alterations in DNA is oxidative damage.
  • Excited-oxygen species such as hydrogen peroxide, hydroxyl radicals, and superoxide radicals arise during irradiation or as a byproduct of aerobic metabolism.
  • Cells possess an elaborate defense system to destroy these reactive species, including enzymes such as catalase and superoxide dismutase.
  • MnSODs manganese-containing superoxide dismutases
  • FeSODs iron-containing superoxide dismutases
  • CuZnSODs unrelated copper and zinc superoxide dismutases
  • Amino acid sequences indicate that MnSODs and FeSODs from diverse sources are closely related, whereas CuZnSODs are not structurally similar. However, all of these SODs catalyze the same process and do so with comparable efficiency.
  • MnSODs and FeSODs are active only with the native metal at the active site. All SODs described to date are multimeric.
  • Mammalian extracellular fluids contain a tetrameric glycosylated CuZnSOD.
  • a group of stable nitroxide free radicals have been shown to possess metal-independent superoxide dismutase- like activity. Unlike SOD, these compounds are of relatively low molecular weight and readily penetrate into the cell.
  • R is an unpaired electron, a cation such as a physiologically acceptable metal ion, hydrogen or a protective group,- and the a and or' carbons are substituted such that when R is an unpaired electron the compound is a stable nitroxide (nitroxyl radical) .
  • a cation such as a physiologically acceptable metal ion, hydrogen or a protective group
  • the a and or' carbons are substituted such that when R is an unpaired electron the compound is a stable nitroxide (nitroxyl radical) .
  • nitroxide nitroxyl radical
  • Preferred are compounds where the ⁇ and ⁇ 1 carbons are substituted to form 5- or 6-membered ring structures. They are useful for preventing and treating ischemic cell damage in mammals. Examples of these nitroxides are 4-hydroxy-2,2, 6, 6- tetramethyl-piperidinyloxyl (TEMPOL) and 4-amino-2,2, 6,6- tetramethyl-piperidinoxyl (TEM
  • Mitchell et al . , WO 91/13619 (1991) discloses compositions containing a metal independent nitroxide compound.
  • Mitchell et al . , 1990 reports SOD mimetic activity in spiro [cyclohexane-1,2' - (4' ,4' - dimethyloxazolidine-3' -oxyl) ] (CHD) and 2-ethyl-2,4,4- trimethyl oxazolidine-3-oxyl (OXANO) .
  • Jarabak and Harvey, 1993, and Samuni et al . , 1991 report SOD activity in 3- carboxy-2,2,5,5-tetramethylproxyl(PCA) .
  • Fridovich et al. U.S. Patent No. 5,227,405 discloses superoxide dismutase-like activity in certain manganese desferroxamine complexes. Riley et al . , 1994, reports SOD mimetic activity for other manganese macrocyclic ligand complexes, including the Mn(II) complex of the macrocyclic ligand, 1,4,7, 10, 13-pentaaxacyclopentadecane.
  • Baudry et al . , 1993 report SOD mimetic activity in several salen-manganese complexes, including both neutral and cationic complexes. Kitajima et al . , 1993, reports monomeric(benzoato) manganese (II) SOD mimetic complexes, such as Mn(OBz) (hydroxytris (3,5-diisopropyl-1- pyrazolyl)borate and Mn(Obz) (3,5-diisopropyl- pyrazole) (hydroxytris (3,5-diisopropyl-l-pyrazolyl)borate. Schechinger et al . , 1988, reports SOD mimic activity in a Cu(II) desferritiocin complex.
  • SOD mimetics that have been identified are metal (including Cu*, Fe 2+ and Mn 2+ ) complexes of pyrimines.
  • metal including Cu*, Fe 2+ and Mn 2+
  • Itami et al reports a complex containing Fe 2* and L-2 (2-pyridyl) -l-pyrroline-5-carboxylic acid.
  • Weiss et al reports a complex containing Fe 2* and L-2 (2-pyridyl) -l-pyrroline-5-carboxylic acid.
  • NO-SOD, N0 2 -SOD, and nitrosylated which possess the ability to detoxify superoxide by catalytic or non-catalytic redox mechanisms enhances the effectiveness of NO therapy because the SOD or compound with SOD-like activity eliminates superoxides which would otherwise consume the administered NO, particularly in its free charged and uncharged species.
  • Nitric oxide and SOD or compounds with SOD-like activity each interact with superoxide, producing peroxynitrite and hydrogen peroxide, respectively.
  • the degradation products of peroxynitrite and hydrogen peroxide can each be toxic depending on the location and conditions under which they are formed, particularly as their concentration increases.
  • the adducts of the invention make it possible to deliver nitric oxide in a non-toxic form.
  • their use enhances the effectiveness of superoxide scavenging by combining nitric oxide inactivation of superoxide with protection of the nitric oxide by the compounds with SOD-like activity.
  • the present invention provides a compound comprising a superoxide oxidant or reductant to which is directly or indirectly linked an NO or N0 2 group. More particularly the invention provides compounds having the formula:
  • R is a moiety that oxidizes and/or reduces superoxide to oxygen and/or hydrogen peroxide under physiological conditions; X is S, N, 0 or C, and D is NO or N0 2 .
  • R can be a superoxide dismutase enzyme.
  • R can be a functional group with an unpaired electron, such as a nitroxide.
  • R can also be a complex of a transition metal and a macrocyclic ligand that dismutes superoxide under physiological conditions.
  • the invention further provides a compound comprising a superoxide oxidant or reductant to which is directly or indirectly linked (i) an NO or N0 2 group and (ii) a specific binding ligand.
  • a specific binding ligand is specifically bindable with a cell surface or extracellular matrix specific binding partner. It is also preferred that the superoxide oxidant or reductant is linked to the NO or N0 2 group through the specific binding ligand.
  • the compounds of the invention are useful for preventing and treating ischemic cell damage in mammals, particularly humans.
  • the invention provides a method of protecting cells from superoxide radical-induced toxicity by contacting the cells with a protective amount of the compounds of the invention.
  • the invention provides for combined or concurrent therapies by administering a superoxide dismutase, superoxide dismutase mimetic or other compound that oxidizes and/or reduces superoxide to oxygen and/or hydrogen peroxide in combination with a nitric oxide adduct.
  • This aspect further provides methods and pharmaceutical preparations comprising such nitric oxide adducts and compounds of the invention.
  • Figure 1 illustrates a synthesis pathway for preparation of a compound of formula IA.
  • Figure 2 illustrates a synthesis pathway for preparation of a compound of formula IIA.
  • Figure 3 illustrates a synthesis pathway for preparation of a compound of formula IIIA.
  • Figure 4 illustrates a synthesis pathway for preparation of a compound of formula IVA.
  • Figure 5A illustrates a synthesis pathway for preparation of a compound of formula VA.
  • Figure 5B illustrates a synthesis pathway for preparation of a compound of formula VB.
  • Figure 6 illustrates a synthesis pathway for preparation of a compound of formula VIA.
  • Figure 7 illustrates a synthesis pathway for preparation of a compound of formula VIIA.
  • the invention provides a compound comprising a superoxide oxidant or reductant to which is directly or indirectly linked an NO or N0 2 group.
  • One embodiment of the invention provides compounds having the formula: D-X-R wherein R is a moiety that oxidizes and/or reduces superoxide to oxygen and/or hydrogen peroxide under physiological conditions; X is S, N, O or C; and D is NO or N0 2 .
  • R can be for example, any of the superoxide dismutase enzymes or mimetics.
  • lower alkyl refers to a branched or straight chain alkyl groups comprising one to ten carbon atoms, including methyl, ethyl, propyl, isopropyl, n- butyl, t-butyl, neopentyl and the like.
  • alkoxy refers to R-O- wherein R is lower alkyl as defined above.
  • Representative examples of alkoxy groups include methoxy, ethoxy, t-butoxy and the like.
  • alkoxyalkyl refers to an alkoxy group as previously defined appended to an alkyl group as previously defined.
  • alkoxyalkyl include, but are not limited to, methoxymethyl, methoxyethyl, isopropoxymethyl and the like.
  • *!amino refers to -NH 2 .
  • dialkylamino refers to (R 12 )N- wherein each R 12 is independently a lower alkyl, for example dimethylamino, diethylamino, methyl propylamino and the like.
  • nitro refers to the group -N0 2 .
  • nitroso refers to the group -NO.
  • hydroxyl or “hydroxy” as used herein refers to the group -OH.
  • cyano refers to the group -CN.
  • N,N-dialkylcarbamoyl refers to R 12 (R 12 )N-C(0)0- wherein each R 12 is independently a lower alkyl, for example dimethylamino, diethylamino, methyl propylamino and the like.
  • N-alkylcarbamoyl refers to R 12 HN- C(0)0- wherein R 12 is a lower alkyl, for example methylamino, ethylamino, propylamino and the like.
  • aryl refers to a mono- or bicyclic carbocyclic ring system having one or two aromatic rings including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl and the like.
  • Aryl groups can be unsubstituted or substituted with one, two or three substituents independently selected from the group consisting of lower alkyl, haloalkyl, alkoxy, amino, alkylamino, dialkylamino, hydroxy, halo, and nitro.
  • substituted aryl groups include tetrafluorophenyl and pentafluorophenyl.
  • arylthio refers to R 13 S- wherein R 13 is aryl.
  • alkanoyl refers to R 12 C(0)- wherein R 12 is a lower alkyl.
  • carboxyl refers to the group -COOH.
  • arylakyl refers to a lower alkyl radical to which is appended an aryl group.
  • Representative arylalkyl groups include benzyl, phenylethyl, hydroxybenzyl, fluorobenzyl, fluorophenylethyl and the like.
  • cycloalkyl refers to an alicyclic group comprising from 3 to 7 carbon atoms including, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.
  • halogen or halo as used herein refers to I, Br, Cl, or F.
  • haloalkyl refers to a lower alkyl radical, as defined above, bearing at least one halogen substituent, for example, chloromethyl, fluoroethyl or trifluoromethyl and the like.
  • heteroaryl refers to a mono-or bi-cyclic ring system containing one or two aromatic rings and containing at least one nitrogen, oxygen, or sulfur in an aromatic ring.
  • Heteroaryl groups can be unsubstituted or substituted with one, two, or three substituents independently selected from the group consisting of lower alkyl, haloalkyl, alkoxy, amino, alkylamino, dialkylamino, hydroxy, halo and nitro.
  • heteroaryl groups include, but are not limited to, pyridine, pyrazine, pyrimidine, pyridazine, pyrazole, triazole, thiazole, isothiazole, benzothiazole, benzoxazole, thiadiazole, oxazole, pyrrole, imidazole, and isoxazole.
  • heterocyclic ring refers to any 3-, 4-, 5-, 6- or 7-membered nonaromatic ring containing at least one nitrogen atom which is bonded to an atom which is not part of the heterocyclic ring.
  • the heterocyclic ring may also contain one additional heteroatom which can be nitrogen, oxygen, or sulfur.
  • the invention also provides nitrosylated and nitrated derivatives of compounds having the formula:
  • B is an unpaired electron, a cation such as a physiologically acceptable metal ion, hydrogen or a protective group; and at least one of the a and ⁇ ' carbons is directly or indirectly linked to an NO or N0 2 group.
  • a and ⁇ 1 carbons are substituted to form 5- or 6-membered ring structures.
  • n is an integer from 0 to 4.
  • R 1 , R 1' , R 2 and R 2' are each independently selected from the group consisting of:
  • Q a in which a is as defined above, is located on the nitrogen-containing ring at one or more positions not substituted by R 1 , R 1' , R 2 , or R 2 ' and is selected from the group consisting of:
  • R 1 , R 1' , R 2 , and R 2' are as defined above;
  • Z is selected from the group consisting of:
  • R s a complex of a transition metal and a macrocyclic ligand, which complex converts superoxide to oxygen and/or hydrogen peroxide under physiological conditions;
  • X is S, N, O or C; and
  • D is NO or N0 2 .
  • R 6 - [R 7 -X-D] a wherein R 6 is a complex of a transition metal and a macrocyclic ligand that dismutes superoxide under physiological conditions; R 7 is a bond or a linking group; and a, X and D are defined as above.
  • M is iron, copper or manganese
  • L is a suitable organic or inorganic ligand or organic or inorganic charge neutralizing anion which is derived from any monodentate or polydentate coordinating ligand or ligand system or the corresponding anion thereof (for example acetic acid acetate anion, alcohol or alkoxide anion)
  • R 7 , a, n, X and D are as defined above.
  • M, L, R 7 , a, n, X and D are as defined above.
  • Another embodiment provides such compounds wherein the macrocyclic ligand is a porphyrin, such as tetrakis (4-benzoic acid) porphyrin.
  • R 8 is a bond, -® . -@ . -@N * . , -@ NO, . -@ SO,H , -@ *>r > ⁇ @ u -® V wherein U is a halogen and V is an alkyl group and wherein ⁇ - indicates bonding to R 9 at any position and ⁇ indicates bonding to R 9 and the substituent at any position; and
  • R 10 is -X-D, -CH 2 -CH 2 -C-X-D, - (CH 2 ) 1 -X-D, -NH-NO or NO " -NO,
  • Another embodiment of the invention provides a compound comprising a superoxide oxidant or reductant to which is directly or indirectly linked (i) an NO or N0 2 group and (ii) a specific binding ligand.
  • the specific binding ligand is specifically bindable with a cell surface or extracellular matrix specific binding partner. It is also preferred that the superoxide oxidant or reductant is linked to the NO or N0 group through the specific binding ligand.
  • Compounds of the invention which have one or more asymmetric carbon atoms may exist as the optically pure enantiomers, pure diastereomers, mixtures of enantiomers, mixtures of diastereomers, racemic mixtures of enantiomers, diastereomeric racemates or mixtures of diastereomeric racemates.
  • the present invention contemplates and includes within its scope all such isomers and mixtures thereof.
  • the nitrosylation of proteins is described in detail in WO93/09806, published 27 May 1993, and the methods of protein nitrosylation described and exemplified there are also applicable to the superoxide dismutases in accordance with the present invention.
  • Mono S-nitrosylation is best achieved by incubating peptides and proteins (in deionized water in an equimolar concentration of acidified nitrite (final concentration 0.5 N HCI) for a period of 1-30 minutes. The incubation time depends on the efficiency of nitrosation and the tolerance of the protein. Nitrosation can also be achieved with a variety of other nitrosating agents including compounds such as S-nitroso-cysteine, S-nitroso-glutathione and related alkyl nitrites. These compounds are to be used when the peptide or protein does not tolerate harsh acidic conditions.
  • polynitrosylated superoxide dismutase proteins themselves it is particularly preferred polynitro ⁇ ylated. Synthesis of polynitrosated peptides and proteins can be achieved in several ways.
  • the peptide or protein is reduced in 100-1000 molar excess dithiothreitol for 30-60 minutes. This exposes intramolecular thiols.
  • the peptide or protein is separated from dithiothreitol by gel filtration (G-25) .
  • the protein is then exposed to increasing concentrations of acidified nitrite (0.5 N HCI) in relative excess over protein.
  • acidified nitrite 0.5 N HCI
  • Complementary measurements of Saville indicate when S-nitrosation is complete. For example, with albumin, this procedure leads to approximately 20 intramolecular S-NO derivatives.
  • the protein can be treated with thiolating agent such as N-acetyl homocy ⁇ teine thiolactone.
  • the derivatized protein can then be S-nitrosated by exposure to acidified nitrite. Exposure to increasing concentrations of nitrite with complementary measurements of Saville can be used to ascertain when S-nitrosation is maximal. Alternatively, thiol groups can be quantified on the protein using standard methodologies and then the protein treated with a stoichiometric concentration of acidified nitrite (0.5 N HCI) .
  • nucleophilic functional groups other than thiol
  • Polynitrosation of nucleophilic functional groups can be achieved when proteins are incubated with excess acidified nitrite.
  • the order of protein reactivity is tyrosine followed by amines on residues such as trytophan. Amide linkages are less reactive. Accordingly, many NO groups can be added to proteins by simply incubating the protein with high excess acidified nitrite. These experiments are performed in 0.5 normal HCI with incubations of approximately one hour. 15 N NMR can be used to determine where the addition (or substitution) by NO takes place.
  • nitrosation can be achieved by exposure to authentic nitric oxide gas under anaerobic conditions in the presence of trace quantities of metals such as copper or silver ion.
  • metals such as copper or silver ion.
  • nitrosation proteins should be incubated in at least 5 atmospheres of NO gas for several hours. Incubation time is protein specific. This can lead to NO attachment to a variety of protein bases. Best characterized reactions involve primary amines. This mechanism provides a pathway to sustain N-nitrosation reactions without deamination. Specifically, exposure to acidified nitrite would otherwise lead to deamination of primary amines whereas this method leads to formation of N- hydroxynitrosamines with potent bioactivity. Similar substitutions at other basic centers also occur. These and other common methods to effect nitrosation are described in the literature, c. f . , D.H.L. Williams, Nitrosation. Cambridge University Press, Cambridge (1988) .
  • Nitroxide compounds of formula I wherein R 1 R 1' , R 2 , R 2' and Q are as defined above can be prepared according to the reaction scheme depicted in Figure 1, in which a substituted piperidine is representative of the saturated nitrogen-containing heterocyclic ring as defined above.
  • 2,2, 6, 6-Tetramethyl-4-piperidinol with the ring nitrogen protected, for example as the t-butoxycarbonyl (t-Boc) derivative is converted to a thiol of formula 2 using a suitable hydroxyl activating agent such as paratoluenesulfonyl chloride or trifluoromethanesulfonic anhydride in the presence of a base (such as triethylamine) , followed by treatment with a suitable sulfur nucleophile such as sodium hydrosulfide.
  • a suitable hydroxyl activating agent such as paratoluenesulfonyl chloride or trifluoromethanesulfonic anhydride
  • a base such as triethylamine
  • sulfur nucleophile such as sodium hydrosulfide.
  • the thiol compound of formula 2 is, in turn, protected as a thiocarbamate by reacting it with a suitable isocyanate such as methoxymethyl isocyanate to afford
  • Deprotection of the ring nitrogen of the compound of formula 3 (t-butoxycarbonyl groups are typically cleaved by treatment with a strong acid such as trifluoroacetic acid) and then oxidizing the resulting amine by treatment with a mild oxidizing agent such as 3-chloroperoxybenzoic acid or sodium tungstate in the presence of aqueous hydrogen peroxide affords the compound of formula 4.
  • the thiol of the compound of formula 4 is deprotected to afford the compound of formula 5.
  • N-methoxymethyl-carbamates are typically cleaved by hydrolysis with a solution of aqueous base. The thiol moiety is then nitrosated using a suitable mild nitrosylating agent such as sodium nitrite in aqueous hydrochloric acid to afford a nitrosothiol of formula IA.
  • Nitroxide compounds of formula II wherein R 1 , R 1' , R 2 R 2' , and Z are defined as above can be prepared according to the reaction scheme depicted in Figure 2, in which a five membered substituted oxazolidine-1-oxyl is representative of the saturated nitrogen-containing heterocyclic ring as defined above.
  • a compound of formula 6 with the thiol moiety protected for example as the S- (N-methoxymethylcarbamate) , wherein a and R 1 are defined as above, is converted to the compound of formula 7 by reacting it with 2-amino-2-methyl-1-propanol in a suitable solvent such as benzene and heating to reflux in the presence of a suitable acid catalyst such as paratoluenesulfonic acid with the azeotropic removal of water.
  • the oxazolidine of formula 7 is converted to the oxazolidine-1-oxyl of formula 8 by treatment with a mild oxidizing agent such as 3-chloroperoxybenzoic acid or sodium tungstate in the presence of aqueous hydrogen peroxide.
  • the compound of formula 8 is then treated with a suitable reagent for removing the thiol protecting group.
  • Treatment with mild aqueous base is the preferred method for removing a S- (N-methoxymethyl-carbamate group.
  • the thiol is then nitrosylated with a mild nitrosating agent, for example, by treatment with sodium nitrite in aqueous hydrochloric acid to afford the compound of formula IIA.
  • Azamacrocycles of formula III wherein R 7 , M, D, a, X, and L are as defined above, and L 1 and L 2 are independently selected from L can be prepared according to the reaction scheme depicted in Figure 3, in which manganese complexes of nitrogen-containing fifteen-membered macrocyclic ligands containing one or two nitrosylated thiols are representative of the transition metal complexes as defined above.
  • a thiol- containing ⁇ -amino acid of formula 9 with the nitrogen and sulfur moieties protected, wherein a, P 1 , and P 2 are defined as above, is activated at the carboxylate with a suitable activating agent such that, when treated with ethylene diamine, it forms the amide of formula 10.
  • Preferred protecting groups for the ⁇ -amino acid of formula 9 are a t-Boc for the amine and a benzyl group for the thiol.
  • one method of activation is formation of the mixed anhydride by reaction of the acid with isobutylchloroformate in the presence of a base such as N-methylmorpholine and subsequent addition of the amine nucleophile.
  • the amide of formula 10 is converted to the triamine of formula 11 by removing the nitrogen protecting group and then reducing the amide functionality.
  • cleavage of a t-butoxycarbamate group is accomplished with a strong acid such as trifluoroacetic acid in methylene chloride or 4N hydrochloric acid in dioxane.
  • the amide is then reduced with a reducing agent such as lithium aluminum hydride or borane in an aprotic solvent such as ether or THF.
  • a reducing agent such as lithium aluminum hydride or borane in an aprotic solvent such as ether or THF.
  • the compound of formula 11 is then converted to the compound of formula 12 by treatment with excess paratoluenesulfonyl chloride in the presence of a non nucleophilic base such as triethylamine or pyridine.
  • Treatment of a compound of formula 12 with a strong base such as sodium hydride in an anhydrous aprotic solvent such as THF or DMF to form the dianion and then reacting the dianion with ethylene carbonate affords a compound of formula 13.
  • the compound of formula 15 is converted to the compound of formula 16 by the removing the amine and sulfur protecting groups.
  • Paratoluenensulfonamides and thiobenzyl ethers can be cleaved with sodium in liquid ammonia. Nitrosation of the thiol group(s) in a compound of formula 16 using a stoichiometric amount of suitable mild nitrosating agent such as sodium nitrite in aqueous hydrochloric acid or t-butyl nitrite in chloroform affords a compound of formula 17. The compound of formula 17 is then converted to the complex of formula IIIA by reaction with a suitable manganese II compound under essentially anaerobic conditions.
  • suitable mild nitrosating agent such as sodium nitrite in aqueous hydrochloric acid or t-butyl nitrite in chloroform
  • Azamacrocycles of formula IV wherein R 7 , M, D, a, X, L 1 , and L 2 are defined as above, can be prepared according to the reaction scheme depicted in Figure 4, in which manganese complexes of nitrogen-containing sixteen-memberedmacrocyclic ligands containing one or two nitrosylated thiol ⁇ are representative of the transition metal complexes as defined above.
  • a suitable hydroxyl activating agent such as paratoluenesulfonyl chloride or trifluoromethanesufonic anhydride
  • a base such as triethylamine
  • R 11 is as defined as above
  • the compound of formula 19 is converted to the compound of formula 20 by the removing the amine and sulfur protecting groups.
  • paratolunensulfonamides and thiobenzyl ethers can be cleaved with sodium in liquid ammonia.
  • a suitable mild nitrosating agent such as a stoichiometric quantity of sodium nitrite in aqueous hydrochloric acid or t-butyl nitrite in chloroform affords a compound of formula 21.
  • the compound of formula 21 is then converted to the complex of formula IVA by reaction with a suitable manganese II compound under essentially anaerobic conditions.
  • Azamacrocycles of formula V in which M, L, R , R , and R 10 are defined as above can be prepared according to the reaction scheme depicted in Figure 5, in which manganese complexes of porphyrin rings are representative of the transition metal complexes as defined above and wherein w is OH, SH, NH 2 or NHR° and x, c, and p 2 are defined as above.
  • T is an activated carbonyl-containing substituent selected from the group consisting of a mixed anhydride, an acid chloride, an isocyanate, or a chloroformate, and c and P 2 are defined as above, to afford compound 24.
  • the thiol protecting group is removed (aqueous base is used to remove S- (N-methoxymethyl- carbamate) groups) to give a compound 25.
  • the thiol moiety is nitrosated with a suitable mild nitrosylating agent such as sodium nitrite in aqueous hydrochloric acid or -butyl nitrite to afford compound 26.
  • Compound 26 is then converted to the complex of formula VA by reacting with a manganese II compound under aerobic conditions.
  • a thiol containing acid with the thiol moiety protected as the S- (N-methoxymethylcarbamate) is converted to the acid chloride 23a with oxalyl chloride and a catalytic amount of DMF and 23a is reacted with 27 in an aprotic solvent such as DMF or THF to afford the amide of formula 28a.
  • the thiol containing acid with the thiol moiety protected as the S(N-methoxymethyl ⁇ carbamate) is converted to the acyl azide by treatment with diphenylphosphoryl azide and this intermediate is heated to catalyze the Curtius rearrangement to the corresponding isocyanate 23b.
  • Treatment of 23b with a compound of formula 27 affords a urea of formula 28b.
  • the thiol containing acid with the thiol moiety protected as the S- (N-methoxymethyl-carbamate) is reduced to the alcohol with borane.
  • Treatment of the alcohol with one equivalent of phosgene affords the chloroformate 23c which is then reacted with a compound of formula 27 to produce the carbamate of formula 28c.
  • the thiol protecting group is cleaved (aqueous base is used to remove S-(N-methoxymethyl-carbamate) groups) and the thiol moiety is nitrosated with a suitable mild nitrosylating agent such as sodium nitrite in aqueous hydrochloric acid or t-butyl nitrite in chloroform to afford the compound of formula 29.
  • a suitable mild nitrosylating agent such as sodium nitrite in aqueous hydrochloric acid or t-butyl nitrite in chloroform to afford the compound of formula 29.
  • the compound of formula 29 is then converted to the complex of formula VIA by reacting with a manganese II compound under aerobic conditions.
  • one method of activation of the carboxylate group is formation of the mixed anhydride by reaction of the acid with isobutylchloroformate in the presence of a base such as N-methylmopholine and subsequent addition of ammonia affords the compound of formula 30.
  • the amide of formula 30 is converted to the diamine of formula 31 by removing the nitrogen protecting group and then reducing the amide functionality.
  • cleavage of a t-butoxycarbamate groups is accomplished with a strong acid such as trifluoroacetic acid in methylene chloride or 4N hydrochloric acid in dioxane.
  • the amide is then reduced with a reducing agent such as lithium aluminum hydride or borane in an aprotic solvent such as ether or THF.
  • a reducing agent such as lithium aluminum hydride or borane
  • an aprotic solvent such as ether or THF.
  • the compound of formula 33 is converted to the compound of formula 34 by removing the ⁇ ulfur protecting group and nitro ⁇ ylating the free thiol.
  • thiobenzyl group ⁇ can be cleaved with sodium in liquid ammonia and nitrosation of the thiol group is accomplished using a suitable mild nitrosating agent such as sodium nitrite in aqueous hydrochloric acid or dinitrogen tetroxide in chloroform.
  • the compound of formula 34 is then converted into the complex of formula VIIA by reacting with a manganese II compound under
  • 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 R is a higher alkyl group than that indicated, or where the tosyl group ⁇ are other nitrogen or oxygen protecting groups or wherein the 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.
  • Activity of the compounds or complexes of the present invention which catalyze the dismutation of ⁇ uperoxide can be demon ⁇ trated u ⁇ ing the stopped-flow kinetic analysis technique as described in Riley et al . , Anal . Biochem. , 196: 344-349 (1991) , which is incorporated by reference herein.
  • Stopped-flow kinetic analysis is an accurate and direct method for quantitatively monitoring the decay rates of superoxide in water.
  • the stopped-flow kinetic analysis is suitable for screening compounds for SOD activity and activity of the compounds or complexes of the present invention, as shown by stopped-flow analysis, correlate to treating the disease states and disorders identified herein.
  • the compounds or complexes of the present invention are novel and can be utilized to treat numerous inflammatory disease states and disorders.
  • reperfusion injury to an ischemic organ e . g. , reperfusion injury to the ischemic myocardium, myocardial infarction, inflammatory bowel disease, rheumatoid arthritis, osteoarthritis, hypertension, psoriasis, organ transplant rejections, organ preservation, impotence, radiation-induced injury, asthma, atherosclerosis, thrombosis, platelet aggregation, metastasis, influenza, stroke, burns, trauma, acute pancreatitis, pyelonephritis, hepatitis, autoimmune disea ⁇ e ⁇ , insulin-dependent diabetes mellitus, disseminated intravascular coagulation, fatty embolism, adult and infantile respiratory distre ⁇ , carcinogenesis and hemorrhages in neonates.
  • the invention also relates to pharmaceutical compositions containing the nitroso-SOD or nitrosylated compounds with SOD-like activity as well as nitro-SOD or nitrated compounds with SOD-like activity, together with a pharmaceutically acceptable carrier.
  • the compounds of the invention can be administered as the sole active pharmaceutical agent, they can also be used in combination or concurrently with one or more compounds which are known to be effective against the specific disease state that one is targeting for treatment.
  • Particularly contemplated in this respect are nitric oxide adducts, such as are discu ⁇ ed below.
  • the invention provides that the ⁇ uperoxide oxidants or reductants which have not been linked to an NO or N0 2 group can be administered in combination or concurrently with nitric oxide or nitric oxide adducts.
  • this aspect pharmaceutical composition comprising a superoxide oxidant or reductant and a nitric oxide adduct in a pharmaceutically acceptable carrier.
  • the superoxide oxidant or reductant can be a superoxide dismutase, for example a manganese-containing superoxide dismutase, an iron-containing superoxide dismutase or a copper and zinc-containing superoxide dismuta ⁇ e.
  • the ⁇ uperoxide oxidant or reductant can be a superoxide dismuta ⁇ e mimetic.
  • these can include, for example, those having the formula a I ⁇ 1
  • R is an unpaired electron, a cation such as a physiologically acceptable metal ion, hydrogen or a protective group; and the a and ⁇ 1 carbons are substituted such that when R is an unpaired electron the compound is a stable nitroxide.
  • superoxide dismutase mimetics include: metal independent nitroxide compounds; spiro[cyclohexane-1,2' - (4' ,4' -dimethyloxazolidine-3' -oxyl) ] , 2-ethyl-2,4,4-trimethyl oxazolidine-3-oxyl; 3-carboxy- 2 , 2, 5, 5-tetramethylproxyl; manganese desferroxamine complexes; macrocyclic ligand ⁇ such as 1,4,7,10,13- pentaaxacyclopentadecane; salen-manganese complexes; monomeric(benzoato) manganese (II) SOD mimetic complexes; Cu(II) desferritiocin complexes; diSchiff base coordinated copper complexes; and metal complexes of a pyrimine.
  • nitric oxide and compounds referred to herein as ''nitric oxide adducts that release nitric oxide or otherwise directly or indirectly deliver or transfer nitric oxide to a site of its activity, such as on a cell membrane, in vivo.
  • nitric oxide encompasses uncharged nitric oxide (NO*) and charged nitric oxide species, particularly including nitrosonium ion(NO * ) and nitroxyl ion(NO " ) .
  • nitric oxide can be provided by gaseous nitric oxide.
  • the nitric oxide releasing, delivering or transferring compounds having the structure X-NO wherein X is a nitric oxide releasing, delivering or transferring moiety, include any and all such compounds which provide nitric oxide to its intended site of action in a form active for their intended purpose.
  • the term "nitric oxide adducts" encompasses any of such nitric oxide releasing, delivering or transferring compounds, including, for example, S-nitrosothiol ⁇ , S-nitroso amino acids, S-nitroso-polypeptides, nitrites and nitrosoamines. It is contemplated that any or all of these "nitric oxide adducts" can be mono- or poly- nitrosylated at a variety of naturally susceptible or artificially provided binding sites for nitric oxide.
  • S- nitrosothiols which are compounds that include at least one -S-NO group.
  • Such compounds include S-nitroso-polypeptides (the term "polypeptide” includes proteins and also polyamino acids that do not pos ⁇ ess an ascertained biological function, and derivatives thereof) ; S-nitrosylated amino acids (including natural and synthetic amino acids and their stereoisomers and racemic mixtures and derivatives thereof) ; S-nitro ⁇ ated sugars, S-nitrosated-modified and unmodified oligonucleotides (preferably of at least 5, and more particularly 5-200, nucleotides) ; and an S-nitrosated hydrocarbon where the hydrocarbon can be a branched or unbranched, and saturated or unsaturated aliphatic hydrocarbon, or an aromatic hydrocarbon; S-nitroso hydrocarbon ⁇ having one or more substituent groups in addition to the S-nitroso group; and heterocyclic compounds.
  • S-nitroso amino acids where the nitroso group is linked to a sulfur group of a sulfur-containing amino acid or derivative thereof.
  • such compounds include the following: S-nitroso-N-acetylcysteine, S-nitro ⁇ o-captopril, S-nitro ⁇ o-homocy ⁇ teine, S-nitro ⁇ o-cysteine and S-nitroso- glutathione.
  • Suitable S-nitrosylated proteins include thiol-containing proteins (where the NO group is attached to one or more ⁇ ulfur group on an amino acid or amino acid derivative thereof) from various functional classes including enzymes, such as tissue- type plasminogen activator(TPA) and cathepsin B; transport protein ⁇ , such as lipoproteins, heme proteins such as hemoglobin and serum albumin; and biologically protective proteins, such, as the immunoglobulins and the cytokines.
  • TPA tissue- type plasminogen activator
  • cathepsin B transport protein ⁇ , such as lipoproteins, heme proteins such as hemoglobin and serum albumin
  • biologically protective proteins such, as the immunoglobulins and the cytokines.
  • nitrosylated proteins are described in PCT Publ. Applic. No. WO 93/09806, published May 27, 1993. Examples include polynitrosylated albumin where multiple thiol or other nucleophilic centers in the protein are modified.
  • S-nitrosothiols include those having the structures:
  • x equals 2 to 20 and Y is selected from the group consi ⁇ ting of halo, alkoxy, cyano, carboxamido, cycloalkyl, arylalkoxy, lower alkylsulfinyl, arylthio, alkylamino, dialkylamino, hydroxy, carbamoyl, N-alkylcarbamoyl, N,N-dialkylcarbamoyl, amino, hydroxyl, carboxyl, hydrogen, nitro and aryl.
  • S-nitroso-ACE inhibitors S-nitroso- angioten ⁇ in converting enzyme inhibitor ⁇
  • R 14 is hydroxy, NH 2 , NHR 24 , NR 4 R 2S , or lower alkoxy, wherein R 24 and R 2S are alkyl, or aryl, or arylalkyl;
  • R 15 is hydrogen, alkyl, arylalkyl, amino, guanidino, NHR 26 , N ⁇ R 26 R 27 ⁇ W erein R 26 and R 27 are methyl or alkanoyl;
  • R 6 is hydrogen, hydroxy, C ⁇ -C alkoxy, phenoxy, or lower alkyl
  • R 17 is hydrogen, alkyl or arylalkyl
  • m i ⁇ 1 to 3 n is 0 to 4 .
  • S-nitroso-ACE inhibitors include N-acetyl- S-nitro ⁇ o-D-cysteinyl -L-proline , N-acetyl-S-nitroso-D, L- cy ⁇ teinyl -L-proline, 1- [4-amino-2- (S-nitro ⁇ o) mercaptomethyl butanoyl] -L-proline , 1- [2 -hexanoyl] -L-proline , 1- [5-- guanidino-2 - (S-nitro ⁇ o) mercaptomethyl-pentanoyl] -L-proline , 1- [5-amino-2- (S-nitro ⁇ o) mercaptomethyl-pentanoyl] -4-hydroxy- L-proline , 1- [5 -guanidino-2 - (S-nitro ⁇ o) mercaptomethyl - pentanoyl] -4 -hydr oxy -L-proline
  • S-nitroso-ACE inhibitors include those having the following structure ⁇ :
  • R 28 is selected from hydrogen, alkyl, arylalkyl, and salt forming ion;
  • R 18 and R 19 are independently aelected from hydrogen, halogen, alkyl, alkoxy, halo substituted lower alkyl, nitro, and S0 2 NH 2 ;
  • G is -C- or -S-
  • R 20 i ⁇ hydrogen, lower alkyl, halo ⁇ ub ⁇ tituted lower alkyl, hydroxy ⁇ ub ⁇ tituted lower alkyl, -(CH 2 ) q -N (lower alkyl) 2 or -(CH 2 ) q -NH 2 and q is one, two, three or four; and
  • R 29 is hydrogen, lower alkyl, alkoxy, halogen or hydroxy and h is as defined above.
  • Additional suitable compounds include those having the following structures:
  • the S-nitroso-ACE inhibitors can be prepared by various methods of synthesis.
  • Acid ⁇ which may be u ⁇ ed for thi ⁇ purpose include aqueous sulfuric, acetic and hydrochloric acids.
  • Thiol precursors are prepared as described in the following: U.S. Pat. Nos.
  • Such compounds include O- nitroso-polypeptides (the term "polypeptide” includes proteins and also polyamino acids that do not pos ⁇ e ⁇ an ascertained biological function, and derivatives thereof) ; O-nitrosylated amino acids (including natural and synthetic amino acids and their stereoisomers and racemic mixtures and derivatives thereof) ; O-nitrosated sugars; O-nitrosated-modified and unmodified oligonucleotides (preferably of at least 5, and more particularly 5-200, nucleotides) ; and an O-nitro ⁇ ated hydrocarbon where the hydrocarbon can be a branched or unbranched, saturated or unsaturated aliphatic hydrocarbon, or an aromatic hydrocarbon; 0-nitroso hydrocarbons having one or more sub ⁇ tituent groups in addition to the O-nitro ⁇ o group; and heterocyclic compounds.
  • O- nitroso-polypeptides the term “polypeptide” includes proteins and also polyamino acids that do not pos ⁇ e
  • NO adducts Another group of such NO adducts is the nitrites which have an -O-NO group wherein R is a protein, polypeptide, amino acid, branched or unbranched and saturated or unsaturated alkyl, aryl or a heterocyclic.
  • R is a protein, polypeptide, amino acid, branched or unbranched and saturated or unsaturated alkyl, aryl or a heterocyclic.
  • a preferred example is the nitosylated form of isosorbide.
  • Compound ⁇ in thi ⁇ group form S-nitro ⁇ othiol intermediates in vivo in the recipient human or other animal to be treated and can therefore include any structurally analogous precursor R-O-NO of the S-nitrosothiols described above.
  • N-nitrosoamines which are compounds that include at least one -N-NO group.
  • Such compounds include N-nitroso-polypeptide ⁇ (the term "polypeptide” include ⁇ protein ⁇ and al ⁇ o polyamino acid ⁇ that do not po ⁇ ess an ascertained biological function, and derivatives thereof) ; N-nitrosylated amino acids (including natural and synthetic amino acids and their stereoisomer ⁇ and racemic mixture ⁇ ) ; N-nitrosated sugars; N-nitrosated-modified and unmodified oligonucleotides (preferably of at least 5, and more particularly 5-200, nucleotides) ; and an N- nitro ⁇ ated hydrocarbon where the hydrocarbon can be a branched or unbranched, and saturated or unsaturated aliphatic hydrocarbon, or an aromatic hydrocarbon; N-nitroso hydrocarbons having one or more substituent groups in addition to the N-nitroso group; and heterocyclic compounds.
  • C-nitroso compound ⁇ that include at least one -C-NO group.
  • Such compounds include C-nitroso-polypeptide ⁇ (the term "polypeptide” include ⁇ proteins and also polyamino acid ⁇ that do not possess an ascertained biological function, and derivatives thereof) ; C-nitrosylated amino acids (including natural and synthetic amino acids and their ⁇ tereoi ⁇ omer ⁇ and racemic mixture ⁇ ) ; C-nitro ⁇ ated ⁇ ugar ⁇ ; C-nitrosated-modified and unmodified oligonucleotides (preferably of at least 5, and more particularly 5-200, nucleotides) ; and a C-nitrosated hydrocarbon where the hydrocarbon can be a branched or unbranched, and saturated or un ⁇ aturated aliphatic hydrocarbon, or an aromatic hydrocarbon; C-nitro ⁇ o hydrocarbon ⁇ having one or more ⁇ ubstituent groups in addition to the C-nitroso group; and heterocyclic compound ⁇ .
  • polypeptides include proteins and also polyamino acids that do not possess an ascertained biological function, and derivatives thereof
  • amino acids including natural and synthetic amino acids and their ⁇ tereoi ⁇ omers and racemic mixtures and derivatives thereof
  • sugars modified and unmodified oligonucleotides (preferably of at least 5, and more particularly 5-200, nucleotides)
  • hydrocarbon where the hydrocarbon can be a branched or unbranched, and saturated or unsaturated aliphatic hydrocarbon, or an aromatic hydrocarbon; hydrocarbons having one or more substituent groups; and heterocyclic compounds.
  • a preferred example is nitroglycerin.
  • Another group of such NO adducts is the nitroso-metal compounds which have the structure (R) n -A-M- (NO) x .
  • R includes polypeptides (the term "polypeptide” includes proteins and also polyamino acids that do not posse ⁇ an a ⁇ certained biological function, and derivatives thereof) ; amino acids (including natural and synthetic amino acids and their ⁇ tereoi ⁇ omer ⁇ and racemic mixture ⁇ and derivatives thereof) ; ⁇ ugar ⁇ ; modified and unmodified oligonucleotide ⁇ (preferably of at lea ⁇ t 5, and more particularly 5-200, nucleotidea) ; and a hydrocarbon where the hydrocarbon can be a branched or unbranched, and saturated or unsaturated aliphatic hydrocarbon, or an aromatic hydrocarbon; hydrocarbons having one or more substituent groups in addition to the A-nitroso group; and heterocyclic compounds.
  • A is S, O, or N
  • n and x are each integers independently selected from 1, 2 and 3
  • M is a metal, preferably a transition metal.
  • Preferred metals include iron, copper, manganese, cobalt, selenium and luthidium. Also contemplated are N-nitro ⁇ ylated metal centers such as nitroprusside.
  • R include ⁇ polypeptides (the term "polypeptide” includes proteins and also polyamino acids that do not possess an ascertained biological function, and derivatives thereof) ; amino acids (including natural and synthetic amino acids and their ⁇ tereoi ⁇ omers and racemic mixtures and derivative ⁇ thereof) ; ⁇ ugars; modified and unmodified oligonucleotides (preferably of at least 5, and more particularly 5-200, nucleotides); and a hydrocarbon where the hydrocarbon can be a branched or unbranched, and saturated or unsaturated aliphatic hydrocarbon, or an aromatic hydrocarbon; hydrocarbons having one or more sub ⁇ tituent groups; and heterocyclic compounds.
  • R is preferably a nucleophilic (basic) moiety.
  • M+ is a metal cation, such
  • thionitrates which have the structure R-(S) x -N0 wherein x is an integer of at least 2.
  • R is as described above for the S-nitro ⁇ othiol ⁇ .
  • dithiols wherein x is 2.
  • Particularly preferred are those compounds where R is a polypeptide or hydrocarbon and a pair or pairs of thiols are ⁇ ufficiently ⁇ tructurally proximate, i.e. vicinal, that the pair of thiol ⁇ will be reduced to a di ⁇ ulfide.
  • Tho ⁇ e compound ⁇ which form di ⁇ ulfide ⁇ pecies release nitroxyl ion(NO " ) and uncharged nitric oxide(NO «) .
  • Those compounds where the thiol groups are not sufficiently close to form disulfide bridges generally only provide nitric oxide as the NO " form not as the uncharged NO* form.
  • the invention further provides various embodiments of pharmaceutical preparations.
  • 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 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 admini ⁇ tration may also involve the use of transdermal administration such as transdermal patches or iontophoresi ⁇ devices.
  • parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection and infusion techniques.
  • injectable preparations for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to the known art using suitable disper ⁇ ing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or ⁇ uspension in a nontoxic parenterally acceptable diluent or ⁇ olvent, for example, a ⁇ a ⁇ olution in 1, 3-butanediol.
  • a nontoxic parenterally acceptable diluent or ⁇ olvent for example, a ⁇ a ⁇ olution in 1, 3-butanediol.
  • the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic ⁇ odium chloride solution.
  • sterile, fixed oil ⁇ are conventionally employed an a solvent or su ⁇ pending medium.
  • any bland fixed oil may be employed including synthetic mono- or diglycerides .
  • fatty acids such as oleic acid find use in the preparation of injectables.
  • Suppositorie ⁇ for rectal admini ⁇ tration of the drug can be prepared by mixing the drug with a ⁇ uitable nonirritating excipient ⁇ uch a ⁇ cocoa butter and polyethylene glycol ⁇ which are ⁇ olid at ordinary temperature ⁇ but liquid at the rectal temperature and will therefore melt in the rectum and release the drug.
  • a ⁇ uitable nonirritating excipient ⁇ uch a ⁇ cocoa butter and polyethylene glycol ⁇ which are ⁇ olid at ordinary temperature ⁇ 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 capsule ⁇ , tablet ⁇ , pill ⁇ , powder ⁇ , granule ⁇ and gel ⁇ .
  • the active compound may be admixed with at least one inert diluent such as sucrose lactose or starch.
  • Such do ⁇ age form ⁇ may also comprise, a ⁇ in normal practice, additional ⁇ ub ⁇ tances other than inert diluents, e.g., lubricating agents such as magnesium stearate.
  • the dosage form ⁇ may al ⁇ o compri ⁇ e buffering agent ⁇ . Tablet ⁇ and pill ⁇ can additionally be prepared with enteric coating ⁇ .
  • 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 composition ⁇ may also comprise adjuvants, ⁇ uch as wetting agents, emulsifying and suspending agents, and sweetening, flavoring, and perfuming agents.
  • the dosage regimen for treating a di ⁇ ea ⁇ e condition with the compound ⁇ 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 sy ⁇ tem i ⁇ utilized and whether the compound is administered a ⁇ part of a drug combination. Thu ⁇ , the dosage regimen actually employed may vary widely and therefore may deviate from the preferred dosage regimen set forth herein.
  • Total daily dose administered to a host in ⁇ ingle 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 amount ⁇ of submultiples thereof to make up the daily do ⁇ e.
  • Cited Literature An and Hsie, Environ. Mol. Mutagen (US), 23:101-109 (1994) An and Hsie, Afutat. Res., 289:215-222 (1993) An and Hsie, Mutat. Res. (NL) , 270:161-115 (1992) Baudry et al., Biochem. Biophys. Res. Comm., 152:964-968 (1993)

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Abstract

Composé constitué d'un oxydant ou d'un réducteur de superoxyde auquel est directement ou indirectement lié un groupe NO ou NO2. L'invention concerne en particulier les composés de formule D-X-R, dans lesquels R est une fraction oxydant et/ou réduisant le superoxyde en oxygène et/ou en peroxyde d'hydrogène dans les conditions physiologiques; X est S, N, O ou C et D est NO ou NO2. R peut être une fonctionnalité comprenant un électron non apparié, un cation tel qu'un ion métal physiologiquement tolérable, hydrogène ou un groupe protecteur, ou R peut être un complexe d'un métal de transition et d'un ligand macrocyclique opérant une dismutation du superoxyde dans les conditions physiologiques. Ces composés peuvent être utilisés seuls ou en association ou concurremment avec d'autres agents thérapeutiques, en particulier les composés d'addition d'oxyde nitrique. De plus, selon l'invention, les oxydants ou réducteurs de superoxydes n'ayant pas de liaison à un groupe NO ou NO2 peuvent être administrés en association ou concurremment avec de l'oxyde nitrique ou des composés d'addition d'oxyde nitrique. Ces composés sont utiles à la prévention des dommages aux cellules par les superoxydes et au traitement des troubles inflammatoires chez les mammifères, en particulier chez l'homme.
PCT/US1996/008406 1995-06-05 1996-06-03 Oxydants et reducteurs nitrosyles et nitres de superoxydes WO1996039409A1 (fr)

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WO1999033823A1 (fr) * 1997-12-23 1999-07-08 Nycomed Imaging As Chelateurs liberant de l'oxyde nitrique et leur emploi a des fins therapeutiques
JP2001527072A (ja) * 1997-12-23 2001-12-25 ニユコメド・イメージング・アクシエセルカペト 一酸化窒素を放出するキレート化剤およびその治療上の使用
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US6759430B2 (en) 1998-01-22 2004-07-06 Oxon Medica Inc. Piperidine and pyrrolidine derivatives comprising a nitric oxide donor for treating stress
US7445641B1 (en) 1999-05-27 2008-11-04 Pharmacia Corporation Biomaterials modified with superoxide dismutase mimics
WO2000072893A3 (fr) * 1999-05-27 2001-08-30 Monsanto Co Biomateriaux modifies par des mimetiques de superoxyde bismuthase
WO2001032202A1 (fr) * 1999-11-03 2001-05-10 Aga Ab Utilisation de monoxyde d'azote pour le traitement d'une constriction du conduit aerien
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US7378438B2 (en) 2002-04-19 2008-05-27 Yissum Research Development Company Of The Hebrew University Of Jerusalem Beta-agonist compounds comprising nitric oxide donor groups and reactive oxygen species scavenger groups and their use in the treatment of respiratory disorders
US7432369B2 (en) 2004-03-29 2008-10-07 Inotek Pharmaceuticals Corporation Pyridyl-substituted porphyrin compounds and methods of use thereof
EP3189836A3 (fr) * 2009-08-19 2017-09-27 Mordechai Chevion Complexes desferrioxamine-métal pour le traitement de troubles liés à l'immunité
WO2013005216A1 (fr) * 2011-07-05 2013-01-10 Radikal Therapeutics Inc. Compositions et méthodes de traitement d'une lésion d'ischémie-reperfusion rénale
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