MXPA06002114A - Mitoquinone derivatives used as mitochondrially targeted antioxidants - Google Patents

Abstract

This invention relates to pharmaceutically acceptable amphiphilic antioxidant compounds, compositions and dosage forms comprising said compounds, and methods and uses reliant on said compounds.The exemplified compounds are all mitoquinone derivatives, being methoxyphenyl alkyl triphenylphosphonium or methoxy dioxocyclohexadiene alkyl triphenylphosphonium derivatives. The compounds, compositions, dosage forms, uses and methods are useful in, for example, the treatment of diseases or conditions associated with oxidative stress.

Description

M1TOQUINONE DERIVATIVES USED AS ANTIOXIDANTS DIRECTED TO THE MITOCHONDRIA FIELD OF THE INVENTION The invention relates to amphiphilic antioxidant compounds having a lipophilic cationic group, and to the synthesis, formulation and physicochemical properties of said compounds that favor their use, for example as drugs.

BACKGROUND OF THE INVENTION Oxidative stress contributes to several human degenerative diseases associated with aging, such as Parkinson's disease and Alzheimer's disease, as well as Huntington's chorea and Friedreich's ataxia, and non-specific damages that accumulate with age. It also contributes to inflammation and injury of ischemic-reperfused tissue in stroke and heart attack and also during organ transplantation and surgery. To prevent the damage caused by oxidative stress several antioxidant therapies have been developed. However, most of these are not directed to the interior of the cells and therefore their efficacy is not optimal. In addition, many of these antioxidants have unfavorable physicochemical properties that limit, for example, their bioavailability and their ability to penetrate the target organ to exert a therapeutic effect. Mitochondria are intracellular organelles responsible for energy metabolism. Consequently, mitochondrial defects are particularly harmful for neutral and muscular tissues that have high energy demands. They are also the main source of free radicals and reactive oxygen species that cause oxidative stress within most cells. Therefore, applicants believe that the selective supply of antioxidants to the mitochondria will be more effective than using non-targeted antioxidants. Accordingly, the present invention is directed to the provision of antioxidants that can be directed to the mitochondria. The lipophilic cations can accumulate in the mitochondrial matrix due to their positive charge (Rottenberg, 1979, Methods Enzymol 55, 547, Chen, 1988, Ann Rev Cell Biol 4, 155). These ions accumulate as long as they are sufficiently lipophilic to cover the positive charge or delocalize it over a large surface area, also provided that there is no active effusion route and the cation is not metabolized or immediately toxic to a cell. The approach of the invention is, therefore, an approach by which it is possible to use the ability of the mitochondria to concentrate specific lipophilic cations to take bound antioxidants, in order to direct the antioxidant to the main source of free radicals and the Reactive oxygen species that cause oxidative stress. Examples of antioxidant compounds that show good antioxidant activity in vivo, but which exhibit deficient antioxidant functionality with respect to the target compartment in vivo, include coenzyme Q (CoQ) and debone. These two compounds have low bioavailability and should be administered at very high dosages to be effective, and therefore have low therapeutic efficacy with respect to the dose administered. The present authors consider, without being limited to any theory, that for an antioxidant compound its in vitro or ex vivo activity (such as for example antioxidant activity or mitochondrial accumulation), in no way is the sole determinant of functionality or antioxidant efficacy in vivo (such as for example therapeutic efficacy). Although it is true that to be useful as antioxidant compounds directed to the mltochondria of the present invention, an antioxidant compound must exhibit adequate antioxidant activity in vitro or ex vivo, to be effective in vivo the antioxidant compound targeted to the mltochondria must exhibit other properties desirable physicochemicals, such as for example suitable bioavailability, localization or adequate distribution within the objective mitochondria or adequate stability. The present authors believe, without wishing to be bound by any theory, that the anti-oxidant compounds of the present invention directed to the mitochondria exhibit advantageous antioxidant functionality, including bioavailability, or mitochondrial targeting and accumulation in vivo, at least in part by virtue of their physicochemical properties, such as for example its amphiphilic character, its structure or physical dimensions, or its hydrophobic character or low or moderate partition coefficient. Said compounds are therefore therapeutically effective at low dosages compared to other antioxidant compounds. In the patent of E.U. No. 6331532, by reference to exemplifications of the compounds mitoquinol and mitoquinone (collectively referred to herein as mitoquinone / mitoquinol), the prospectus of mitochondrial targeting of an antioxidant portion based on a lipophilic cation covalently coupled to the antioxidant portion is described. The compound exemplified there (despite the generalization of bridge length) is the compound mitoquinone of the formula: with a carbon bridge length of 10 (that is, bridge of Cío). In its reduced form, mitochonol also has the Cio-bridge. It has been found that mitochonon / mitoquinol, despite having an excellent antioxidant activity and targeting and accumulation in the mitochondria in vitro and in vivo, is a bit unstable as the bromide salt. The present authors have also found that the physicochemical properties of mitochonon / mitoquinol, described in the US patent. No. 6,331, 532, are unsuitable for a pharmaceutical formulation, for example when administration is oral or parenteral, or when it is sought to direct the compound to the mitochondria in tissues of internal organs (eg brain, heart, liver or other organs). Examples of compounds of the present invention are suitable for pharmaceutical formulation. They may be in a different form from a crystalline or solid form, but are suscepfible from forming a solid form by mixing with other agents, such as for example vehicles, excipients, complexing agents or other additives and the like, such as by example cyclodextrins. Advantageously, said agents are pharmaceutically acceptable. The present authors have determined the desirability of offering examples of the amphiphilic antioxidant compounds of the present invention directed to the mitochondria, with their positive charge in association with a suitable anion to thereby provide the compound as a neutral general salt form, which includes products solid or crystalline. However, the present authors have found that in such salt forms it is better to avoid some salt-forming anions, since they exhibit reactivity against the antioxidant compound, for example against the antioxidant portion, the binding portion, or the lipophilic cationic portion, or they produce a breakdown of the antioxidant portion. Other salt-forming anions are considered pharmaceutically undesirable. For example, nitrate portions are generally considered inappropriate by pharmaceutical companies for being pharmaceutically or environmentally unacceptable, while the present inventors have found that hydrogen bromide, frequently used in the formation of salt of said compounds, has properties nucleophilic which can lead to a reactivity with the antioxidant portion, for example a breakdown of a methyl group of an antioxidant portion of the compound of general formula (II) hereof, or some general reduction in the stability of the compound as a whole. For example, the present authors have determined that the mitochonon bromide salt is a little unstable. Therefore, the present authors consider that salt forms of anti-oxidants directed to mitochondria, including salt forms in liquid, solid or crystalline form, are better associated with an anion or similar portion that is not nucleophilic, or which exhibits no reactivity against any of the portions comprising the antioxidant compound or complex. It is also preferable that the anion be pharmaceutically acceptable.

OBJECT OF THE INVENTION Accordingly, an object of the present invention is to provide pharmaceutically acceptable amphiphilic antioxidant compounds, and compositions, dosage forms and methods based on said compounds, which are useful for example in the treatment of diseases or conditions associated with oxidative stress, or for provide the public with a useful choice.

BRIEF DESCRIPTION OF THE INVENTION In a first aspect, the present invention consists of a compound comprising a lipophilic cationic moiety bound by a binding portion to an antioxidant portion, and an anionic complement to said cationic moiety, wherein the cationic species are capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement does not exhibit reactivity against the antioxidant portion, the cationic portion or the binding portion. In one embodiment, the antioxidant portion is a quinone or a quinol. In other embodiments, the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives, chain-breaking antioxidants, including butylated hydroxyanisole, butylated hydroxytoluene, radical scavengers in general, including modified fullerenes, spin traps that They include derivatives of 5,5-dimethylpyrroline N-oxide, fer-butylnitrosobenzene, ter-nitrosobenzene, a-phenyl-fer-butylnitrone, and related compounds.

In one embodiment, the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation. In one embodiment, the compound has the general formula I: or its quinol form, wherein Ri, R2 and R3, which may be equal or different, are selected from alkyl portions of C1 to C5 (optionally substituted) or H, and wherein n is an integer from about 2 to about 20, and where Z is a non-reactive anion. Preferably, Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. Preferably, each C of the bridge (C) n is saturated. In a preferred embodiment, the compound has the formula: II or its quinol form, wherein Z is a non-nucleophilic anion. Most preferably, the compound has the formula: (lll) In another aspect, the invention provides a pharmaceutical composition comprising or including a compound comprising a lipophilic cationic moiety bound by a binding moiety to an antioxidant moiety, and an anionic moiety for said cationic moiety, where the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion, or the binding portion. In one embodiment, the antioxidant portion is a quinone or a quinol. In other embodiments, the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives, chain breaking antioxidants including butylated hydroxyanisole, butylated hydroxytoluene, radical scavengers in general including modified fullerenes, spin traps including derivatives of 5,5-dimethylpyrroline N-oxide, fer-butylnitrosobenzene, urea-nitrosobenzene, α-phenyl-fer-butylnitrone, and related compounds. In one embodiment, the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation. In one embodiment, the compound has the general formula l: or its quinol form, wherein Ri, R2 and 3, which may be the same or different, are selected from alkyl portions of C1 to C5 (optionally substituted) or H, and wherein n is an integer from 2 to 20, approximately , and where Z is a non-reactive anion. Preferably, Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. Preferably, each C of the bridge (C) n is saturated. In a further embodiment, the compound has the formula: or its quinol form, where Z is a non-nucleophilic anion. In a further embodiment, the composition comprises a compound having the formula II or its quinol form, wherein Z is a non-nucleophilic anion and wherein the composition comprises cyclodextrin. In several examples, the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10, from about 5: 1 to about 1: 5, from about 4: 1 to about 1: 4, of about 2: 1 at about 1: 2, or about 1: 1; for example, the molar ratio of compound to cyclodextrin is about 1: 2. Most preferably, the composition comprises a compound having the formula: wherein the cyclodextrin is β-cyclodextrin, and preferably the molar ratio of compound to cyclodextrin is about 1: 2. In one embodiment, the pharmaceutical composition is formulated for oral administration. In a further embodiment, the pharmaceutical composition is formulated for parenteral administration. In a further aspect, the present invention provides a unit dose comprising or including a compound comprising a lipophilic cationic moiety bound by a binding portion to a portion of antioxidant, and an anionic complement to said cationic moiety, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion or the binding portion, together with any diluent, carrier or pharmaceutically acceptable excipient. In one embodiment, the antioxidant portion is a quinone or a quinol. In other embodiments, the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives; chain-breaking antioxidants including butylated hydroxyanisole, butylated hydroxytoluene; radical scavengers in general including modified fullerenes, spin traps including derivatives of 5,5-dimethyryl pyrroline N-oxide, fer-butylnitrosobenzene, ione-nitrosobenzene, α-phenyl-ér-butylnitrone; and related compounds. In one embodiment, the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation. In one embodiment, the compound has the general formula I: or its quinol form, wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of C1 to C5 (optionally substituted) or H, and wherein n is an integer from about 2 to about 20 , and where Z is a non-reactive anion. Preferably, Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. Preferably, each C of the bridge (C) n is saturated. In a further embodiment, the compound has the following formula, or its quinol form, wherein Z is a non-nucleophilic anion.

In a further embodiment, the unit dose comprises a compound having the formula II or its quinol form, wherein Z is a non-nucleophilic anion and wherein the composition comprises cyclodextrin. In several examples, the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1:10, from about 5: 1 to about 1: 5, from about 4: 1 to about 1: 4, of about 2: 1. at about 1: 2, or about 1: 1; for example, the molar ratio of compound to cyclodextrin is about 1: 2. Most preferably, the unit dose comprises a compound having the formula: (ffl) wherein the cyclodextrin is β-cyclodextrin, and preferably the molar ratio of compound to cyclodextrin is about 1: 2. In one embodiment, the unit dose is suitable for oral administration. In a further embodiment, the unit dose is suitable for parenteral administration. In a further aspect, the present invention provides a compound or a pharmaceutically acceptable salt thereof, a composition, or a dosage form of the present invention, for use in the prophylaxis or treatment of oxidative stress in a mammal, by administering the compound or its salt to said mammal.

In one embodiment, the compound is a compound of formula II or a pharmaceutically acceptable salt thereof. In another embodiment, said administration is, on the first day, at a dose of about 1.02 to about 2.0 times the daily maintenance dose, followed by administration of the compound or its salt at the daily maintenance dose on subsequent days. Preferably, the salt is methanesulfonate and the compound is combined with cyclodextrin. Most preferably, the compound has the formula: G? Preferably, the cyclodextrin is β-cyclodextrin and preferably the molar ratio of compound to cyclodextrin is about 1: 2. In a further aspect, the present invention provides a compound or a pharmaceutically acceptable salt thereof, a composition, or a dosage form of the present invention, for use in the prophylaxis or treatment of symptoms of aging in a mammal, by administration of the compound or its salt to said mammal. In one embodiment, the compound is a compound of formula II or a pharmaceutically acceptable salt thereof. In another embodiment, said administration is, on the first day, at a dose of about 1.02 to about 2.0 times the daily maintenance dose, followed by administration of the compound or its salt at the daily maintenance dose on subsequent days.

Preferably, the salt is methanesulfonate and the compound is combined with cyclodextrin. Most preferably, the compound has the formula: (HE) Preferably, the cyclodexyrin is β-cyclodextrin and preferably the molar ratio of compound to cyclodextrin is about 1: 2. In a further aspect, the present invention consists of a stable compound comprising a lipophilic cationic moiety bound by a binding portion to an antioxidant portion, and an anionic complement to said cationic moiety, wherein: the cationic species is capable of directing the antioxidant species to the mitochondria, and the anionic complement is not a halogen ion, and the anionic complement is non-nucleophilic, or the anionic complement does not exhibit reactivity against the cationic portion, the binding portion, or the antioxidant portion. In one embodiment, the ampho-oxidant portion is a quinone or a quinol. In other embodiments, the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives; chain-breaking antioxidants including butylated hydroxyanisole, butylated hydroxytoluene; radical scavengers in general including modified fullerenes, spin traps including derivatives of 5,5-dimethylpyroline, N-oxide, urea-butylnitrosobenzene, tert-nitrosobenzene, α-phenyl-fer-butylnitrone; and related compounds.

In one embodiment, the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphon cation. In one embodiment, the compound has the general formula I: or its quinolue form, wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of Ci to C5 (optionally substituted) or H, and wherein n is an integer from about 2 to about 20 , and where Z is a non-reactive anion. Preferably, Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. Preferably, each C of the bridge (C) n is saturated. In a preferred embodiment, the compound has the formula: or its quinol form, where Z is a non-nucleophilic anion. Most preferably, the compound has the formula: In another aspect, the invention provides a pharmaceutical composition comprising or including a stable compound comprising a cationic species which is a lipophilic cationic moiety bound by a binding portion to a portion of antioxidant, and an anionic complement for said cationic moiety. , where: the cationic species is capable of directing the antioxidant species to the mitochondria, and the anionic complement is not a halogen ion, and the anionic complement is not nucleophilic, or the anionic complement does not exhibit reactivity against the cationic portion, the of binding or the antioxidant portion. In one embodiment, the antioxidant portion is a quinone or a quinol. In other embodiments, the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives; chain-breaking antioxidants including butylated hydroxyanisole, butylated hydroxytoluene; radical scavengers in general which include modified fullerenes, spin traps including derivatives of 5,5-dimethylpyrroline N-oxide, urea-butylnitrosobenzene, fer-nitrosobenzene, α-phenyl-fer-butylnitrone; and related compounds. In one embodiment, the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation. In one embodiment, the compound has the general formula I: or its quinolue form, wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of Ci to C5 (optionally substituted) or H, and wherein n is an integer from about 2 to about 20 , and where Z is a non-reactive anion. Preferably, Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. Preferably, each C of the bridge (C) n is saturated. In a further embodiment, the compound has the formula: or its quinol form, where Z is a non-nucleophilic anion. In a further embodiment, the composition comprises a compound having the formula II or its quinol form, wherein Z is a nucleophilic anion and wherein the composition comprises cyclodextrin. In several examples, the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1:10, from about 5: 1 to about 1: 5, from about 4: 1 to about 1: 4, of about 2: 1. at about 1: 2, or about 1: 1; for example, the molar ratio of compound to cyclodextrin is about 1: 2. Most preferably, the composition comprises a compound having the formula: wherein the cyclodextrin is β-cyclodextrin, and preferably the molar ratio of compound to cyclodextrin is about 1: 2. In one embodiment, the pharmaceutical composition is formulated for oral administration. In a further embodiment, the pharmaceutical composition is formulated for parenteral administration. In a further aspect, the present invention consists of a unit dose comprising or including a stable compound comprising a lipophilic cationic moiety bound by a binding portion to a portion of antioxidant, and an anionic complement for said cationic moiety. , together with any pharmaceutically acceptable diluent or carrier or excipient, where: the cationic species is capable of directing the antioxidant species to the mitochondria, and the anionic complement is not a halogen ion, and the anionic complement is not nucleophilic, or the anionic complement does not exhibit reactivity against the cationic portion, the of binding or the antioxidant portion. In one embodiment, the antioxidant portion is a quinone or a quinol. In other embodiments, the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives; chain-breaking antioxidants including butylated hydroxyanisole, butylated hydroxytoluene; radical scavengers in general which include modified fullerenes, spin traps including derivatives of 5,5-dimethylpyrroline N-oxide, ér-butylnitrosobenzene, fer-nitrosobenzene, α-phenyl-tert-butylnitrone; and related compounds. In one embodiment, the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation. In one embodiment, the compound has the general formula l: 1 or its quinol form, wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of C1 to C5 (optionally substituted) or H, and wherein n is an integer from about 2 to about 20 , and where Z is a non-reactive anion. Preferably, Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. Preferably, each C of the bridge (C) n is saturated. In a further embodiment, the compound has the formula: or its quinol form, where Z is a non-nucleophilic anion. In a further embodiment, the unit dose comprises a compound having the formula II or its quinol form, wherein Z is a non-nucleophilic anion and wherein the composition comprises cyclodextrin. In several examples, the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10, from about 5: 1 to about 1: 5, from about 4: 1 to about 1: 4, of about 2: 1 at about 1: 2, or about 1: 1; for example, the molar ratio of compound to cyclodextrin is about 1: 2. Most preferably, the unit dose comprises a compound having the formula: wherein the cyclodextrin is β-cyclodextrin, and preferably the molar ratio of compound to cyclodextrin is about 1: 2.

In one embodiment, the unit dose is suitable for oral administration. In a further embodiment, the unit dose is suitable for parenteral administration. In a further aspect, the present invention consists of a unit dose suitable for oral administration, comprising as active ingredient a compound according to the present invention, the compound being of, or being formulated as, a crystalline form or a non-liquid form . In a further aspect, the present invention consists of a suitable unit dose for parenteral administration, comprising as active ingredient a compound according to the present invention. In a further aspect, the present invention provides a pharmaceutical composition suitable for the treatment of a patient, which would benefit from a reduction of oxidative stress or a reduction of aging symptoms, comprising or including an effective amount of a compound of the present invention in combination with one or more pharmaceutically acceptable vehicles, excipients or diluents. In one embodiment, the compound is a compound of formula I. In one example, the compound is complexed with cyclodextrin. In several examples, the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10, from about 5: 1 to about 1: 5, from about 4: 1 to about 1: 4, of about 2: 1 at about 1: 2, or about 1: 1; for example, the molar ratio of compound to cyclodextrin is about 1: 2. Most preferably, the compound is a compound of formula (III) and the cyclodextrin is β-cyclodextrin, and the preferred molar ratio of compound to cyclodextrin is about 1: 2. In a further aspect, the invention provides a method of reducing oxidative stress in a cell, comprising the step of contacting said cell with a compound of the present invention. In one embodiment, the compound is a compound of formula I. In one example, the compound is complexed with cyclodextrin. In several examples, the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10, from about 5: 1 to about 1: 5, from about 4: 1 to about 1: 4, of about 2: 1 at about 1: 2, or about 1: 1; for example, the molar ratio of compound to cyclodextrin is about 1: 2. Most preferably, the compound is a compound of formula (III) and the cyclodextrin is β-cyclodextrin, and the preferred molar ratio of compound to cyclodextrin is about 1: 2.

In one embodiment, the pharmaceutical composition is formulated for oral administration. In a further embodiment, the pharmaceutical composition is formulated for parenteral administration. In a further aspect, the present invention provides a pharmaceutical composition suitable for the treatment of a patient suffering from or predisposed to suffering from Parkinson's disease, Alzheimer's disease, Huntington's chorea, or Friedreich's ataxia, which comprises or includes a effective amount of a compound of the present invention, in combination with one or more pharmaceutically acceptable vehicles, excipients or diluents. Preferably, said treatment is for a patient suffering or predisposed to undergoing Friedreich's ataxia. In a further aspect, the invention provides a method of therapy or prophylaxis for a patient who would benefit from a reduction of oxidative stress, comprising or including the step of administering to said patient a compound of the present invention. In one embodiment, the compound is a compound of formula I. In one example, the compound is complexed with cyclodextrin. In several examples, the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10, from about 5: 1 to about 1: 5, from about 4: 1 to about 1: 4, of about 2: 1 at about 1: 2, or about 1: 1; for example, the molar ratio of compound to cyclodextrin is about 1: 2. Most preferably, the compound is a compound of formula (III) and the cyclodextrin is β-cyclodextrin, and the preferred molar ratio of compound to cyclodextrin is about 1: 2. In one embodiment, said administration is oral administration. In another embodiment, said administration is parenteral administration. In another aspect, the invention provides a method of therapy or prophylaxis for a patient who would benefit from a reduction of oxidative stress or a reduction in the symptoms of aging, comprising the step of administering to the patient a compound of the present invention. . In another aspect, the present invention provides a method of therapy or prophylaxis for a patient suffering from or predisposed to suffering from Parkinson's disease, Alzheimer's disease, Huntington's chorea, or Friedreich's ataxia, which comprises or includes the passage of administering to said patient a compound of the present invention. Preferably, the method of therapy or prophylaxis is for a patient suffering or predisposed to undergoing Friedreich's ataxia. In another aspect, the invention provides a method for reducing oxidative stress in a cell, comprising the step of administering to the cell a compound of the present invention. In another aspect, the invention provides the use of a compound as described above, in the preparation or manufacture of a medicament, unit dose or pharmaceutical composition effective to reduce oxidizing stress in a patient. In another aspect, the invention provides the use of a compound as described above, in the preparation or manufacture of a medicament, unit dose or pharmaceutical composition effective to reduce the symptoms of aging in a patient. In a further aspect, the invention provides the use of a compound of the present invention in the preparation or manufacture of a medicament, unit dose or pharmaceutical composition effective for the treatment or prophylaxis of a patient, who suffers from or is predisposed to suffer the disease of Parkinson's disease, Alzheimer's disease, Huntington's chorea, or Friedreich's ataxia, which comprises or includes the step of administering to said patient a compound of the present invention. Preferably, the medicament, unit dose or pharmaceutical composition is effective for use in the treatment or prophylaxis of a patient suffering from or predisposed to undergoing Friedrich's ataxia. In another aspect, the invention provides for the use of a compound as described above, in the preparation or manufacture of a medicament, unit dose or pharmaceutical composition, effective for use in the reduction of oxidative stress in a cell.

Preferably, said preparation or manufacture is with one or more other materials, preferably pharmaceutically acceptable diluents, excipients or vehicles. In a further aspect, the present invention consists of a method of synthesizing a compound with a portion or portion of formula I: (or its quinone form), wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of Ci to Cs (optionally substituted), and wherein n is an integer from 2 to 20, said method comprising or comprising mixing with cyclodextrin. Preferably, each C of the bridge (C) n is saturated. In a further aspect, the present invention consists of a method of synthesis of a compound having the formula: said method comprising or comprising mixing with cyclodextrin. In a further aspect, the present invention consists of a method of synthesizing a compound having the formula: essentially as described here. Any disclosure of documents, records, materials, devices, articles or the like, which has been included in the present specification, is only intended to provide a context for the present invention. It is not considered an admission that any or all of these matters form part of the basis of the prior art, nor that they are of general knowledge common in the field relevant to the present invention, as existing prior to the priority date of each claim of this request. In all this specification, it will be understood that the word "comprises" or variations such as "comprising", implies the inclusion of an element, integer or referred step, or group of elements, integers or steps referred to, but not the exclusion of some other element, integer or step, or group of elements, integers or steps. In all this specification, it shall be understood that the term "quinone", used alone or in front of another term to describe the oxidized form of a compound, includes within its scope the reduced form of that compound, that is, the form of quinol. Similarly, the reference to a quinone, for example by structural representation, also includes within its scope the quinol form. Throughout this specification, it will be understood that the term "quinol", used alone or prefixed to another term to describe the reduced form of a compound, includes within its scope the oxidized form of that compound, that is, the quinone form. Similarly, the reference to a quinol, for example by structural representation, also includes within its scope the quinone form. As used herein, the term "or" includes both "and" and "or" as options.

As used herein, the terms "partition coefficient" and "partition coefficient (octanol / water)" refer to the partition coefficient in 1-octanol / phosphate buffer, determined at 25 ° C or 37 ° C ( see Kelso, GF, Porteous, CM, Coulter, CV, Hughes, G. Porteous, WK, Ledgerwood, E. C, Smith, RAJ and Murphy, MP, 2001 J Biol Chem 276, 4588, Smith, RAJ, Porteous, CM , Coulter, CV and Murphy, MP 1999, Eu. J Biochem 263, 709, Smith, RAJ, Porteous, CM, Win, AM and Murphy, MP, 2003, Proc Nat Acad Sci 100, 5407), or the partition coefficient octanol / water calculated using the Advanced Chemistry Development (ACD) Solaris V4.67 software, as described by Jauslin, ML, Wirth, T., Meier, T., and Schoumacher, F., 2002, Hum Mol Genet 11, 3055. As used herein, the phrase "acceptable for pharmaceutical preparation" includes within its meaning not only acceptance as to its pharmaceutical administration, but also with respect to the formulation, for example stability, shelf life, hygroscopicity, preparation and the like acceptable. As used herein, a "non-reactive anion" is an anion that exhibits no reactivity against the antioxidant portion, the lipophilic cation, or the binding portion. For example, if one such portion of the compound comprises a nucleophilic attack target, the anion is non-nucleophilic. Although it was broadly defined above, the invention is not limited to that, but also consists of the modalities whose examples are given below.

In particular, a better understanding of the invention will be obtained with reference to the attached drawings.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 represents the incorporation of amphiphilic antioxidant compounds in the mitochondria, where the incorporation of mitochondrion C10 into an energized mitochondria is shown schematically. Figures 2A-2C represent the synthetic routes for: A, mitoquinone C3; B, Mitoquinone C5, C: C15 mitoquinone. Figures 3A-3F depict the structure of antioxidant compounds of mitochonon and the related compound TPMP. A phospholipid drawn to the same scale is aligned with the anti-oxidant compounds of mitoquinone, to indicate the maximum potential depths of penetration of the side chain of ubiquinol into a flake of a phospholipid blot. A: TPMP. B: Mitoquinone C3. C: mitoquinone C5. D: C10 mitoquinone. E: C15 mitoquinone. F: Phospholipid. Figures 4A-4D present graphs showing the incorporation and binding of antioxidant compounds in the mitochondria, measured using a selective ion electrode. A: Mitoquinone C3. B: Mitoquinone C5. C: C10 mitoquinone. D: mitoquinone C15. In the panels on the left, mitochondria (1 mg protein / ml) were present in the presence of rotenone and then the antioxidant compounds were added, such as five sequential additions of 1 μM (black arrowheads) to calibrate the electrode response. For the panels on the right, the electrodes were first calibrated with five sequential additions of 1 μM (black arrowheads) and then mitochondria (1 mg protein / ml) were added. In all cases succinate was added to generate a membrane potential, and FCCP was added to dissipate it. The data are typical traces of repeated experiments at least 2-3 times. Figures 5A-5D depict graphs showing antioxidant efficacy of antioxidant compounds. A: The mitochondria were energized with succinate (black bars) or by incubation with an ATP regeneration system consisting of ATP, phosphoenol pyruvate and pyruvate kinase (white bars). After a preincubation of 30 seconds with the various analogues of mitochonon, TPMP or vehicle, oxidative stress was induced by adding 50 μM of FeCl2 and 300 μM of H2O2. After 15 minutes of incubation at 37 ° C, lipid peroxidation was estimated by measuring TBARs. The data are means ± amplitude of two independent experiments. The slight protective effect of mitochonon C5 on lipid peroxidation in the presence of ATP is due to the fact that a part of the C5 mitoquinone added from the supply solution was in the form of ubiquinol. B: The mitochondrial membrane potential induced with succinate or with the ATP regeneration system was measured from the accumulation of [3H] TPMP. The data are means ± amplitude of duplicate determinations of a 25 minute incubation. The membrane potentials after a 5 minute incubation were the same (data not shown). C: The concentration dependence of the prevention of accumulation of TBARs with the antioxidant compounds was measured. All incubations were done in the presence of succinate as described for A. The results are expressed as the percentage of inhibition of TBARS formation, taking the value of a sample exposed to FeCl2 / H202 in the absence of mitochonon analogs as 0% of inhibition, and a control sample (without adding FeCl2 / H202) as 100%. The data shown is a typical titration with each concentration determined in triplicate ± SD. D: Cl5o concentrations of prevention of lipid peroxidation. The data are means ± sem, estimated from three independent titrations as shown in C. The statistical significance with respect to the Cl50 of the C3 mitoquinone was determined using the two-tailed Student's t-test: * p < 0.05; ** p < 0.005. Figure 6 presents a graph showing the effect of C10 mitoquinone and C3 mitoquinone on coronary sinus flow. Figure 7 presents a graph showing the effect of mitochondrone C10 and mitochondon C3 on left ventricular diastolic pressure. Figure 8 presents a graph showing the effect of mitochondrone C10 and mitoquinone C3 on heart rate.

Figures 9A and 9B present graphs showing the velocity of left ventricular change. Figures 10A and 10B show graphs showing the effect of mitochondrial C10 and mitochondrone C3 on mitochondrial respiratory function after ischemia. Figure 11 is a graph depicting the stability of pure C10 mitochonone (lot # 3) in clear glass containers at 40 ° C, 75% RH; 25 ° C, 50% RH; and 5 ° C on silica gel. Figure 12 is a graph representing the stability of the C10 mitoquinone (batch No. 4) at 25 ° C, 50% RH. Figure 13 is a graph depicting the stability of the C10 mltoquinone complex and β-cyclodextrin (1: 1) at 4 ° C on silica; at 25 ° C, 50% RH; and at 40 ° C, 75% RH. Figure 14 is a graph depicting the stability of the complex of mitochondone C10 and β-cyclodextrin (1: 2) at 4 ° C on silica; at 25 ° C, 50% RH; and at 40 ° C, 75% RH. Figure 15 is a graph depicting the stability of the complex of mitochondone C10 and β-cyclodextrin (1: 4) at 4 ° C on silica; at 25 ° C, 50% RH; and at 40 ° C, 75% RH. Figure 16 is a graph showing the stability of the mesylate of C10 mitoquinone in water. Figure 17 presents a graph showing the stability of the mitochonone mesylate C10 in 0.01 M HCl.

Figure 18 presents a graph showing the stability of the mitochonone mesylate C10 in IPB, pH 7.4. Figure 19 presents a graph showing the stability of the mitochonone mesylate C10 in 50% MeOH. Figure 20 presents a graph showing the solid state stability of mitochondone mesylate C10 at 40 ° C, 75% RH; at 25 ° C, 50% RH; and at 4 ° C on blue silica gel. Figure 21 presents a graph showing the stability of the mitochondone meslate complex C10 and β-cyclodextrin (1: 2) in water. Figure 22 presents a graph showing the stability of the mitoxulon mesylate complex C10 and β-cyclodexyrin (1: 2) in 0.01 M HCl. Figure 23 presents a graph showing the stability of the cycloquinone mesylate complex C10 and β -cyclodextrin (1: 2) in IPB, pH 7.4. Figure 24 presents a graph showing the stability of the complex of mitochondone mesylate C10 and β-cyclodextrin (1: 2) in 50% MeOH. Figure 25 presents a graph showing the solid state stability of the mitochondone mesylate complex C10 and β-cyclodextrin (1: 2), at 40 ° C, 75% RH; at 25 ° C, 59% RH; and at 4 ° C on blue silica gel. Figure 26 presents a graph showing the stability of the mitochondone mesylate complex C10 and β-cyclodextrin (1: 1) in water.

Figure 27 presents a graph showing the stability of the mitochondrion mesylate complex C10 and β-cyclodextrin (1: 1) in 0.01 M HCl. Figure 28 presents a graph showing the stability of the mitochondrion mesylate complex C10 and β -cyclodextrin (1: 1) in IPB, pH 7.4. Figure 29 presents a graph showing the stability of the complex of mesylate of mitochonon C10 and β-cyclodextrin (1: 1) in 50% methanol. Figure 30 shows a graph showing the solid state stability of the mitochondone mesylate complex C10 and β-cyclodextrin (1: 1) at 40 ° C, 75% RH; at 25 ° C, 50% RH; and at 4 ° C on blue silica gel. Figures 31A and 31B present graphs of the concentration profiles of mitochondrone C10 in the rat plasma with respect to the time, after a single administration via i.v. (A; 10 mg / kg) and oral (B; 50 mg / kg) of mitochondone mesylate C10 in the mitochondone mesylate complex C10 and β-cyclodextrin (1: 2) (n = 5). The pharmacokinetic parameters derived from these data are given in table 11.

DETAILED DESCRIPTION OF THE INVENTION As indicated above, this invention focuses on directing compounds to the mitochondria, mainly for therapy or prophylaxis to reduce oxidative stress. The mitochondrion has a substantial membrane potential of up to 180 mV through its internal membrane (negative inside). Because of this potential, membrane-permeable lipophilic cations accumulate several hundred times within the mitochondrial matrix. Applicants have found that by coupling lipophilic cations (for example the lipophilic triphenylphosphonium cation) with an antioxidant portion, the resulting amphiphilic compound can be delivered to the mitochondrial matrix within intact cells. The antioxidant is then directed to a site of primary production of free radicals and reactive oxygen species within the cell, rather than being randomly dispersed. Now, the applicants have also determined that the properties of the antioxidant compound, such as for example the nature of the antioxidant portion, the physical and chemical characteristics of the binding portion, such as the length or the lipophilic nature of the binding portion , or the nature of the lipophilic cation, contribute to the efficacy of the antioxidant compound in vivo and contribute to the antioxidant functionality of the compound. For the antioxidant compounds of the present invention, the in vivo efficacy may comprise in part suitable bioavailability, adequate stability, adequate pharmacokinetics, suitable antioxidant activity, or proper mitochondrial targeting or accumulation.

In principle, in the formation of the compounds of the invention, any lipophilic cation and any antioxidant, capable of being transported to and through the mitochondrial membrane, and accumulated in or within the mitochondria of intact cells can be used. However, it is preferred that the lipophilic cation is the triphenylphosphonium cation exemplified herein. Other lipophilic cations that can be covalently coupled with the antioxidants according to the present invention include tribencylammonium cations and phosphonium. In some examples of antioxidant compounds of the present invention, the lipophilic cation is coupled to the antioxidant portion by means of a saturated linear carbon chain having from 1 to about 30 carbon atoms, for example from 2 to about 20 atoms of carbon, from about 2 to about 15, from about 3 to about 10, or from about 5 to about 10 carbon atoms. In a particularly preferred example, the linear carbon chain comprises carbon atom. Preferably, the carbon chain is an alkylene group (e.g., CrC20, or CrC15), however, carbon chains which can optionally include one or more double or triple bonds are also within the scope of the invention. Also included are carbon chains that include one or more substituents (such as hydroxyl, carboxylic acid or amide groups), or include one or more side chains or branches, such as those selected from alkyl, alkenyl or alkynyl groups, substituted or not replaced. Also included are carbon chains that comprise more than about 30 carbon atoms, but whose length is equivalent to a saturated linear carbon chain having from 1 to about 30 carbon atoms. It will be appreciated by those skilled in the art that different portions of straight alkylene can be used to covalently couple the amphoteric portion with the lipophilic cation, for example substituted or branched alkyl groups, peptide bonds, and the like. In some embodiments, the lipophilic cation is linked to the antioxidant portion by means of a straight-chain alkylene group having from 1 to 10 carbon atoms, such as for example an ethylene, propylene, butylene, pentylene, or decylene group. Antioxidant portions useful in the present invention include those that require interaction with reducing agents to have antioxidant activity, either for the initial antioxidant activity or to recycle the antioxidant activity, or both. For example, the antioxidant compounds of the present invention which comprise as an active antioxidant portion a quinol portion, can be administered in the form of quinone. To function as an antioxidant, that is, to have antioxidant activity, the quinone must be reduced to the quinol form by interaction with a reducer, such as for example a mitochondrial reducer such as complex II, for the initial antioxidant activity. The subsequent interaction of the oxidized form of the quinone with reducing agents can lead to the recycling of the antioxidant activity.

Other examples of antioxidant portions useful in the present invention include those that already exist as the reduced form and do not require interaction with reducing agents to have an initial antioxidant activity. However, the subsequent interaction of the oxidized form of such amphotericin portions with mitochondria reducers can lead to the recycling of the antioxidant activity. For example, the antioxidant portion vitamin E can be administered in reduced form and thus does not require interaction with reducers to have initial antioxidant activity, but subsequently interacts with reducers such as for example the endogenous deposit of quinone, to thereby recycle the antioxidant activity. Additional examples of antioxidant portions useful in the present invention include those that are not recycled by interaction with mitochondrial reductants. Examples of antioxidant portions useful in the present invention include vitamin E and vitamin E derivativeschain-breaking antioxidants, such as butylated hydroxyanisole, butylated hydroxytoluene, quinols and radical scavengers in general, such as modified fullerenes. In addition, spin traps that react with free radicals to generate stable free radicals can also be used. These will include derivatives of 5,5-dimethylpyrroline N-oxide, tert-butylnitrosobenzene, fer-nitrosobenzene, a-f-enyl-butylnitrone, and related compounds.

Preferred amphoteric compounds, which include those of the general formulas I and II hereof, can be easily prepared, for example by the following reaction: The general synthesis strategy is to heat for several days a precursor containing a suitable leaving group, preferably a precursor of alkyl sulfonyl, bromine or iodine, with more than 1 equivalent of triphenylphosphine, under argon. Then, the phosphonium compound is isolated as its salt. To do this, the product is triturated repeatedly with diethyl ether until a whitish solid remains. This is then dissolved in chloroform or dichloromethane and precipitated with diethyl ether to remove excess triphenylphosphine. This is repeated until the solid is no longer dissolved in the chloroform. At this point, the product recrystallizes several times from a suitable solvent, such as chloroform, acetone, ethyl acetate or higher alcohols. A preferred synthetic method that can be used to prepare a stable form of an antioxidant compound directed to the preferred mitochondria of formula III (also referred to herein as C10 mitochonone mesylate or C10 mitochonone methanesulfonate), is as set forth in Example 1 of the present. It will also be appreciated that by using ion exchange or other known techniques the anion of the antioxidant compound thus prepared can be readily exchanged for another pharmaceutically or pharmacologically acceptable anion, if this is desirable or necessary. Applicants have determined that the stability of the salt form of the antioxidant compound improves when the anion exhibits no reactivity towards the antioxidant portion, the binding portion or the lipophilic cationic portion. For example, in the case of the preferred examples of antioxidant compounds of the invention, the anion is non-nucleophilic. It is also desirable that the anion be a pharmaceutically acceptable anion. It is also preferred that for pharmaceutical formulation, the anion does not exhibit reactivity towards any other agent comprised in the formulation. Examples of non-nucleophilic anions include hexafluoroantimonate, arsenate, or phosphate, or tetraphenylborate, tetra- (perfluorophenyl) borate or other tetrafluoroborates, trifluoromethanesulfonate, aryl, and alkyl sulfonates, such as methanesulfonate and p-toluenesulfonate, and phosphates. Examples of pharmaceutically acceptable anions include halogen ions such as fluoride ion, chloride, bromide ion and iodide ion; anions of salts of inorganic acids, such as nitrate, perchlorate, sulfate, phosphate and carbonate; pharmaceutically acceptable anions of salts of lower alkylsulfonic acid, such as the salts of methanesulfonic acid and ethanesulfonic acid; pharmaceutically acceptable anions of arylsulfonic acid salts, such as the salts of benzenesulfonic acid, 2-naphthalenesulfonic acid and p-toluenesulfonic acid; pharmaceutically acceptable anions of salts of organic acids, such as the salts of trichloroacetic acid, trifluoroacetic acid, hydroxyacetic acid, benzoic acid, mandelic acid, butyric acid, propionic acid, formic acid, fumaric acid, succinic acid, citric acid, acid, tartaric, oxalic acid, maleic acid, acetic acid, malic acid, lactic acid and ascorbic acid; and pharmaceutically acceptable anions of salts of acidic amino acids, such as the salts of glutamic acid and aspartic acid. In the case of preferred examples of antioxidant compounds of the invention, the halogen anion precursor is exchanged for aryl or alkyl sulfonate anions. Examples include, without limitation, benzenesulfonate, p-toluenesulfonate, 2-naphthylenesulfonate, methanesulfonate, ethanesulfonate, propanesulfonate. A particularly preferred anion is the methanesulfonate anion. As described above, an example of an antioxidant compound of the invention wherein the anion is methanesulfonate, is the particularly preferred antioxidant compound of formula III, referred to herein as C10 mitochonone methanesulfonate or C10 mitochonone mesylate. The same general procedure can be used to make a wide range of compounds directed to the mitochondria with different antioxidant portions R attached to the portion or portions of triphenylphosphonium (or other lipophilic cationic portions). These will include a series of vitamin E derivatives in which the length of the bridge that couples the function of vitamin E with the triphenylphosphonium portion (or another lipophilic cationic portion) is varied. Other antioxidants that can be used as R include chain breaking antioxidants, such as butylated hydroxyanisole, butylated hydroxytoluene, quinols and radical scavengers in general such as modified fullerenes. In addition, you can also synthesize spin traps, which react with free radicals to generate stable free radicals. These include derivatives of 5,5-dimethylpyrroline N-oxide, fer-butylnitrosobenzene, fer-nitrosobenzene, a-phenyl-fer-butylnitrone, and related compounds. It will be appreciated that for an antioxidant compound of the present invention, as for any drug, in vitro activity is in no way the sole determinant of functionality or efficacy in vivo. The antioxidant activity of the antioxidant compounds of the present invention can be determined by methods such as those described herein, using for example isolated mitochondria or isolated cells. Although it is true that to be useful as an antioxidant compound directed to the mitochondria of the present invention, an antioxidant compound must exhibit an adequately high antioxidant activity in said tests, to be effective in vivo, the antioxidant compound directed to the mitochondria must exhibit other desirable physico-chemical properties, for example suitable bioavailability, stability or antioxidant functionality.

Examples of antioxidant compounds that exhibit good ampholytic activity but that exhibit poor bioavailability with respect to the target compartment in vivo, include coenzyme Q (CoQ) and idebenone. These two compounds should be administered at very high dosages (eg 0.5-1.2 g) to obtain minimal clinical effects in human patients. Antioxidant compounds directed to the mitochondria exemplary of the present invention exhibit good antioxidant activity and bioavailability, and therefore are effective in vivo at low dosages. In example 11 of this description, the determination of the bioavailability of an amphiphilic antioxidant compound directed to the preferred mitochondria of the present invention, the mitochonone mesylate C10 and a cyclodextrin complex thereof is presented. The present authors consider that the antioxidant compounds of the present invention are effective in directing antioxidant activity to the mitochondria, exhibiting one or more of the additional benefits of being available in crystalline or solid form or being capable of being formulated in a solid form. , of greater stability, greater bioavailability or greater antioxidant functionality. The present authors consider, again without wishing to be limited to any theory, that the physical and chemical characteristics of the antioxidant compounds of the present invention confer to the antioxidant compounds of the present invention preferred characteristics, which thus allow their use in compositions, formulations and methods, among other applications, for which the antioxidant compounds of the prior art may be less suitable given their chemical and physical properties. In some embodiments of the invention, the antioxidant compound is a quinoline derivative of the formula II defined above. For example, a quinol derivative of the invention is the mitochonone compound C10 as defined above (of which the compound of formula III is a specific salt form). A further example of the invention is a compound of formula I in which (C) n is (CH2) s and the quinol portion is the same as that of C10 mitoquinone, referred to herein as C5 mitoquinone (see Figure 3C). A further example of the compound of the invention is a compound of formula I wherein (C) n is (CH2) 3 and the quinol portion is the same as mitochonone C10, which is referred to herein as C3 mitoquinone (see Figure 3B) . Yet another example of a compound of the invention is a compound of formula I wherein (C) n is (CH2) 5 5 and the quinol portion is the same as that of C10 mitoquinone, referred to herein as C15 mitoquinone (see figure 3E). Once the ampholydant compound of the invention is prepared in any pharmaceutically appropriate form, and optionally including pharmaceutically acceptable carriers, excipients, diluents, complexing agents or additives, it will be administered to the patient in need of therapy or prophylaxis. Once administered, the compound will direct antioxidant activity to the mitochondria within the patient's cells.

The antioxidant compounds of the present invention can be administered to patients by oral or parenteral routes of administration. The antioxidant compound must be formulated into a stable and safe pharmaceutical composition for administration to a patient. The composition can be prepared according to conventional methods, by dissolving or suspending an amount of the antioxidant compound in a diluent. The amount is between 0.1 mg and 1000 mg of the amphoteric compound per ml of diluent. A buffer of acetate, phosphate, citrate or glutamate can be added which allows the pH of the final composition to be from 5.0 to 9.5; optionally, a carbohydrate or polyhydric alcohol tonifier and a preservative selected from the group consisting of m-cresol, benzyl alcohol, mephoryl, ethyl-, propyl- and butylparabens and phenol. A sufficient amount of water for injection is used to obtain the desired concentration of the solution. If desired, additional tonicity agents such as sodium chloride, as well as other excipients may also be present. However, said excipients must maintain the overall tonicity of the antioxidant compound. The terms "buffer", "buffer" and "buffered solution", when used to refer to the concentration of the hydrogen ion or pH, refer to the ability of a system, particularly an aqueous solution, to resist a change in pH by adding acid or alkali to it, or when diluted with a solvent. It is characteristic of buffer solutions, which undergo small changes in pH by the addition of acid or base, the presence of a weak acid and a weak acid salt, or a weak base and a weak base salt. An example of the first system is acetic acid and sodium acetate. The change in pH is light as long as the amount of hydroxyl ion added does not exceed the capacity of the buffer system to neutralize it. The stability of the parenteral formulations of the present invention is increased by maintaining the pH of the formulation in the range of about 5.0 to 9.5. Other pH scales include, for example, 5.5 to 9.0, or 6.0 to 8.5, or 6.5 to 8.0, or 7.0 to 7.5. The buffer used in the practice of the present invention is selected from any of the following, for example, an acetate buffer, a phosphate buffer, or a glutamate buffer, with phosphate buffer being most preferred. Vehicles or excipients may also be used to facilitate the administration of the compound. Examples of carriers and excipients include calcium carbonate, calcium phosphate, various sugars such as lactose, glucose or sucrose, or types of starch, cellulose derivatives, gelatin, polyethylene glycols and physiologically compatible solvents. A stabilizer may be included in the present formulation but, importantly, it is not necessary. If included, however, a stabilizer useful in the practice of the present invention is a carbohydrate or a polyhydric alcohol. Polyhydric alcohols include compounds such as sorbitol, mannitol, glycerol and polyethylene glycols (PEGs). The carbohydrates include, for example, mannose, ribose, trehalose, maltose, inositol, lactose, galactose, arabinose or lactose. Suitable stabilizers include, for example, polyhydric alcohols such as sorbitol, mannitol, inositol, glycerol, xylitol, and polypropylene / efilenglycol copolymers, as well as various polyethylene glycols (PEG) of molecular weight 200, 400, 1450, 3350, 4000, 6000 and 8000). The United States Pharmacopeia (USP) states that antimicrobial agents should be added at bacteriostatic or fungistatic concentrations to preparations contained in multiple-dose containers. They must be present at a suitable concentration at the time of use to prevent the multiplication of microorganisms inadvertently introduced into the preparation, while removing a portion of the contents with a hypodermic needle and syringe or using other invasive means for delivery, such as injectors. pen. Antimicrobial agents should be evaluated to ensure compatibility with all other components of the formula, and their activity should be evaluated in the total formula to ensure that a particular agent that is effective in one formulation is not ineffective in another. It is not uncommon to find that a particular agent is effective in one formulation but not in another. In the common pharmaceutical sense, a preservative is a substance that prevents or inhibits microbial growth, and can be added to a pharmaceutical formulation for this purpose to avoid the consequent decomposition of the formulation by microorganisms.

Although the amount of the preservative is not very large, it can nevertheless affect the overall stability of the antioxidant compound. In this way, even the selection of a curator can be difficult. Although the preservative for use in the practice of the present invention may vary from 0.005 to 1.0% (w / v), the preferred scale for each preservative, alone or in combination with another, is: benzyl alcohol (0.1-1.0%) or m-cresol (0.1-0.6%), or phenol (0.1-0.8%), or a combination of methylparaben (0.05-0.25%) and etll- or propyl- or butylparaben (0.005% -0.03%). Parabens are lower alkyl esters of para-hydroxybenzoic acid. A detailed description of each conservator is given in "Remington's Pharmaceufical Sciences" and in "Pharmaceutical Dosage Forms: Parenteral Medications", Vol. 1, 1992, Avis and others. For this purpose, the crystalline dihydrochloride trientine salt can be administered parenterally (including subcutaneous injections, intravenous, intramuscular, intradermal injection, or infusion techniques) or by inhalation atomizer, in unit dose formulations containing vehicles, adjuvants and excipients conventional non-toxic and pharmaceutically acceptable. It may also be convenient to add sodium chloride or other salt to adjust the tonicity of the pharmaceutical formulation, depending on the tonifier selected. However, this is optional and depends on the particular formulation selected. Parenteral formulations should be isotonic or substantially isotonic, otherwise significant irritation and pain would occur at the site of administration.

The desired isotonicity can be achieved using sodium chloride or other pharmaceutically acceptable agents, such as dextrose, boric acid, sodium tartrate, propylene glycol, polyols (such as mannitol and sorbitol), or other inorganic or organic solutes. In general, the composition is isotonic with the subject's blood. If desired, the parenteral formulation can be thickened with a thickening agent such as methylcellulose. The formulation can be prepared in an emulsified form, either water in oil or oil in water. Any of a wide variety of pharmaceutically acceptable emulsifying agents can be employed which include, for example, acacia powder, a nonionic surfactant or an ionic surfactant. It may also be convenient to add suitable dispersing or suspending agents to the pharmaceutical formulation, and these may include for example aqueous suspensions such as synthetic and natural gums, that is, tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone or jelly. The most important vehicle for parenteral products is water. Water of adequate quality for parenteral administration must be prepared by distillation or by reverse osmosis. Only by these means it is possible to properly separate from the water various liquid, gaseous and solid contaminants. Water for injection is the preferred aqueous vehicle for use in the pharmaceutical formulation of the present invention. The water can be purged with nitrogen gas to remove any oxygen or oxygen-free radical from the water. It is possible that other ingredients may be present in the parenteral pharmaceutical formulation of the present invention. Such additional ingredients may include wetting agents, oils (for example, a vegetable oil such as sesame, peanut or olive oil), analgesic agents, emulsifiers, amphotericizers, bulking agents, tonicity modifiers, metal ions, vehicles. oleaginous, proteins (for example, human serum albumin, gelatin or proteins) and a zwitterion (for example, an amino acid such as betaine, taurine, arginine, glycine, lysine and histidine). Said additional ingredients, of course, should not adversely affect the overall stability of the pharmaceutical formulation of the present invention. The containers are also an integral part of the formulation of an injection and can be considered as a component; there is no container that is totally insoluble or that does not affect in any way the liquid that it contains, particularly if the liquid is aqueous. Therefore, the selection of a container for a particular injection should be based on a consideration of the composition of the container, as well as the solution and the treatment to which it will be subjected. To allow the insertion of a needle of a hypodermic syringe into a multiple dose bottle and to provide resealing as soon as the needle is removed, each bottle is sealed with a rubber closure held in place by an aluminum band.

Plugs for glass jars can be used, such as West 4416/50, 4416/50 (coated with Teflon) and 4406/40, Abbott 5139, or any equivalent cap. These plugs pass the plug integrity test in tests using patient use models, for example, the plug can withstand at least about 100 injections. All the components of the pharmaceutical formulation described above are known and described in "Pharmaceutical Dosage Forms: Parenteral Medications ", Vol. 1, 2nd ed., Avis et al, ed., Mercel Dekker, New YorK, N. Y. 1992, which is incorporated in its entirety as a reference herein. The manufacturing process of the above formulation includes the steps of mixing, sterile filtration and filling. The mixing process can include, for example, the dissolution of the ingredients in a specific order, for example first the preservative, followed by the stabilizer or tonicity agent, buffers, and then the antioxidant compound, or dissolving all the ingredients at the same time. which form the parenteral formulation. An example of a method of preparing a formulation for parenteral administration is the dissolution in water of the amphro-oxidant compound form, for example mitochonone mesylate C10-β-cyclodextrin (1: 2), and dissolution of the resulting mixture in a solution phosphate buffer salt. Alternatively, the parenteral formulations of the present invention are prepared by mixing the ingredients following the generally accepted procedures. For example, the selected components can be mixed in a mixer or other standard device to produce a concentrated mixture, which can then be adjusted to the final concentration and viscosity by the addition of water, a thickening agent, a buffer, 5% albumin of human serum, or an additional solute to control the tonicity. Alternatively, the antioxidant compound can be packaged as a dry powder or solid to be reconstituted with a solvent, to produce a parenteral formulation according to the present invention for use at the time of reconstitution. In addition, the manufacturing process can include any suitable sterilization process in developing the parenteral formulation of the present invention. Typical sterilization procedures include filtration, steam (moist heat), dry heat, gases (eg, ethylene oxide, formaldehyde, chlorine dioxide, propylene oxide, beta-propiolactone, ozone, chloropicrin, peracetic acid methyl bromide, and similar), radiation exposure and aseptic handling. Suitable routes of parenteral administration include intravenous, intramuscular, subcutaneous, intradermal, subdermal, intratraumatic, intrathecal, intraperitoneal, and the like. The intravenous route of administration is preferred. The mucosal supply is also admissible. The dose and dosage regimen will depend on the weight and health of the subject.

The pharmaceutically acceptable carriers, excipients, diluents, complexing agents or additives may be chosen in such a way as to, for example, increase the stability of the amphoteric compound, facilitate the synthesis or formulation of the pharmaceutical preparation, or increase the bioavailability of the amphoteric compound. . For example, it is well known that carrier molecules such as cyclodextrin and derivatives thereof have potential as complexing agents capable of altering the physicochemical attributes of the drug molecules. For example, cyclodextrins can stabilize (both thermally and oxidatively), reduce volatility and alter the solubility of the active agents with which they form the complex. Cyclodextrins are cyclic molecules composed of glucopyranose ring units that form toroidal structures. The interior of the cyclodextrin molecule is hydrophobic and the exterior is hydrophilic, making the dextrin molecule soluble in water. The degree of solubility can be altered by replacing the hydroxyl groups on the outside of the cyclodextrin. Similarly, the hydrophobic character of the interior can be altered by substitution, although generally the hydrophobic nature of the interior allows the accommodation of relatively hydrophobic host portions within the cavity. The accommodation of one molecule within another is known as complex formation and the resulting product is referred to as an inclusion complex. Examples of cyclodextrin derivatives include sulfobufilcyclodextrin, maltosylcyclodextrin, hydroxypropylcyclodextrin, and salts thereof. . Example 1 and Example 7 of the present invention describe methods of preparing a pharmaceutically acceptable composition comprising an inclusion complex of an antioxidant compound directed to the mitochondria, in this case mitochondrion C10 in complex with β-clclodextrin. In Example 9 and Example 10, methods of preparing pharmaceutically acceptable compositions comprising an inclusion complex of an antioxidant compound directed to the preferred mitochondrion, mitochonone mesylate C10 in complex with β-cyclodextrin are described. The physicochemical properties, which include for example the pharmaceutical properties of the antioxidant-cyclodextrin compound complex, can be varied, for example by varying the molar ratio of antioxidant compound to cyclodextrin, or by varying the cyclodextrin itself. For example, for the preferred antioxidant compounds of general formula I, the molar ratio of antioxidant compound to cyclodextrin (antioxidant compound: cyclodextrin) can be from about 10: 1 to about 1:10, from about 5: 1 to about 1: 5. , from about 4: 1 to about 1: 4, from about 2: 1 to about 1: 2, or about 1: 1. In a further example, the preferred molar ratio of the exemplary antioxidant compound, C10-mitoquinone, to cyclodextrin, is 1: 2, and the cyclodextrin is β-cyclodextrin. Alternatively, the pharmaceutically appropriate form of the antioxidant compound can be formulated in order to increase its stability and bioavailability. For example, enteric coatings may be applied to the tablets to prevent the release of the antioxidant compound in the stomach, either to reduce the risk of unpleasant side effects or to maintain the stability of the antioxidant compound that would otherwise be subject to exposure degradation. to the gastric medium. Most of the polymers used for this purpose are polyacids that function by virtue of the fact that their solubility in an aqueous medium is pH dependent, and require conditions with a pH higher than that normally found in the stomach. A preferred type of oral controlled release structure is the enteric coating of a solid dosage form. The enteric coatings promote that the compounds remain physically incorporated in the dosage form during a specific period when exposed to gastric juice, but are designed to disintegrate in the intestinal fluid for rapid absorption. The delay of absorption depends on the rate of transfer through the gastrointestinal tract, and thus, the speed of gastric emptying is an important factor. For some administrations, a dosage form of the multiple unit type, such as granules, may be superior to the type of individual units. Therefore, in one embodiment, the antioxidant compounds of the invention can be contained in a multi-unit, enteric-coated dosage form. In a highly preferred embodiment, the dosage form of the antioxidant compound is prepared by producing particles having an amphiphilic compound-enteric coating agent solid, in an inert core material. These granules can result in a prolonged absorption of the amphoteric compound with good bioavailability. Typical enteric coating agents include, without limitation, hydroxypropylmethylcellulose phthalate, methacrylic acid-methacrylic acid ester copolymer, polyvinyl acetalphthalate and cellulose acetate phthalate. Exemplary preferred antioxidant compounds of the present invention, or formulations or complexes thereof, exhibit advantageous pharmaceutical properties. For example, they are easily formulated, they are chemically and physically stable, they are easily soluble in water, they have low hygroscopicity, and they exhibit a good life, useful in storage. Next, the invention will be described in more detail with reference to the following non-limiting experimental section.

Example 1: Synthesis of mitochondon C10 The following discloses a preferred method of synthesis of a preferred stable salt form of the exemplary mitochondrial-directed antioxidant compound, C10-mitoquinone mesylate, and a cyclodextrin complex thereof. STAGE 1 Scheme: Dichloromethane Methanesulfonyl chloride C3% SOaCÍ P.M.338.44 P.M. 114.55 P.M.416.53 Steps: 1. Idebenone (A1, 0.25 kg, 0.74 mol) is dissolved in 2.5 L of DCM reactive grade and then the mixture is cooled to 10 ± 3 ° C under an inert atmosphere. 2. Fracylamine (0.152 kg, 1.5 mol) is added in one portion at room temperature, and the mixture is allowed to rebalance at 10 ± 3 ° C. 3. A solution of methanesulfonyl chloride (0.094 kg, 0.82 mol) in 0.5 L DCM is then gradually added at a rate such that an internal temperature of approximately 10-15 ° C is maintained (the addition ends in this scale). after 75 minutes). 4. The reaction mixture is stirred 15-30 more minutes. 5. I PC is verified with respect to termination by means of TLC (Rf 0.65, 5% ethanol / dichloromethane). 6. The mixture is then washed with water (0.85 L) and saturated aqueous sodium bicarbonate solution (0.85 L). 7. The organic layer evaporates under reduced pressure at 40-45 ° C until a red liquid remains. After drying for a further 2-4 hours at high vacuum and at room temperature, the crude product thus obtained, A2, is used directly in the next step. The yield is unknown since the solvent was trapped in the liquid.

STAGE 2 Scheme: Methanol Sodium borohydride C8H3207S NáBH_j C20H3 O S P.M.416.53 P.M.37.83 P.M.418.55 Steps: 1. Dissolve idebenone mesylate (A2, assume 100% yield from the last step, 0.31 kg, 0.74 mol) in 2L of methanol and then the mixture is cooled to 0-5 ° C under an inert atmosphere. 2. Sodium borohydride (0.03 kg, 0.79 mol) is added in portions, at a suitable speed to ensure that the internal temperature does not exceed 15 ° C. The termination of the reaction will be accompanied by a change of color: red? yellow (on this scale, the addition is completed after 20 minutes). 3. The reaction mixture is stirred for a further 10-30 minutes. 4. CPI for completion is verified by means of TLC (A3, Rf 0.60, ethanol 5% / dichloromethane, A2 Rf 0.65). 5. Then, the mixture is inactivated with 2 L of 2M hydrochloric acid solution and extracted three times with 1.2 L of dichloromethane. 6. Then, the combined organic phase is washed once with 1.2 L of water and dried over anhydrous magnesium sulfate (0.24 kg). 7. The organic phase is then evaporated under reduced pressure at 40-45 ° C, until leaving a yellow / brown syrup. After drying an additional 2-8 hours under high vacuum at room temperature, the crude product thus obtained, A3, 0.304 kg, 98% yield, is used directly in the next step.

STAGE 3 Scheme: Triphenylphosphine C3sH4907PS P.M.418.55 P.M.680.83 AS A4 Steps: 1. Pieces of triphenylphosphine (0.383 kg 1.46 mol) are added to idebenol mesylate (A3, 0.304 kg, 0.73 mol) in a properly sized round bottom flask. 2. Next, the flask is coupled with a rotary evaporator and the contents are heated under vacuum to a bath temperature of 80-85 ° C. 3. The mixture should form a homogeneous molten suspension at this temperature. Once the molten suspension is formed and the degassing is no longer evident, the vacuum is displaced with an inert atmosphere and the mixture is gently rotated in a bath at 80-85 ° C for about 3 days. 4. Verify IPC termination by means of 1 H and 31 P NMR. A minimum of 95% conversion is required before doing the treatment. 5. The mixture is then cooled to almost room temperature and dissolved in 0.8 L of dichloromethane. 6. Then 3.2 L of ethyl acetate is added in portions with gentle heating to precipitate the desired product from excess triphenylphosphine. 7. A small volume of solvent is removed by evaporation under reduced pressure (to remove the DCM) and then the remaining mixture is cooled almost to room temperature and decanted. 8. After, the remaining viscous residue is subjected twice more to the same washing procedure and finally dried at high vacuum to a constant weight, to produce a cinnamon foam, 0.441 kg, 89% yield (note: the product still contained something of solvent, see NMR). The A4 product thus obtained is used directly in the next step.

STAGE 4 Scheme: P.M.680.83 P.M.678.82 A.4 A5 Steps: 1. The crude mitoquinol mesylate salt (0.44 kg, assume 0.65 mol) is dissolved in 6 L of anhydrous DCM and the flask is purged with oxygen. 2. The contents of the flask are stirred vigorously under an atmosphere of oxygen for 30 minutes to ensure saturation of the solvent with the gas. 3. 0.1 L of 0.65 M N02 solution in dry DCM (2 mol% N02) is rapidly added in one portion; The mixture is stirred vigorously under an oxygen atmosphere for 4-8 hours at room temperature. 4. IPC is then verified for termination (by means of 1 H NMR and optionally 3 P NMR).

. If the oxidation is incomplete, add 2 mol% more of N02 as a solution in DCM. This would take the reaction to completion. Verify IPC as above. In this scale, 8 mol% of N02 was required as a solution in DCM for the reaction to reach completion. 6. Then, the solvent is removed by evaporation under reduced pressure to produce a red viscous residue. This residue is dissolved in 2 L of dichloromethane at 40-45 ° C. 7. Then add 3.2 L of ethyl acetate in portions with gentle heating to precipitate the desired product. A small volume of the solvent is removed by evaporation under reduced pressure (to remove the DCM), and the remaining mixture is then cooled to almost room temperature and decanted. 8. Finally, the oily residue is dried under high vacuum to a constant weight to produce a red crystal (419 g, 94% yield). The product A5 thus obtained is used directly in the next step.

STAGE 5 Scheme: Steps: 1. The crude mitoquinone mesylate salt (A5, 0.419 kg) is dissolved in 6 L of water with gentle heating at 40-43 ° C. 2. The beta-cyclodextrin, 1.23 kg, is dissolved separately in 20 L of water, with heating at 60 ° C. 3. These two solutions are cooled to approximately room temperature and combined to form a homogeneous mixture. This solution should be saved to < 5 ° C. 4. This orange solution is then frozen at -20 ° C and lyophilized in batches at constant weight (at least 48 hours). 5. The resulting solid is then crushed gently to form a uniformly free yellow / orange fluid powder (1433 kg). An alternative synthetic method has been performed wherein the oxidation step 3 of step 4 of the synthetic method described above, is made by bubbling oxygen through the solution, indicating that the oxidation reaction can be handled substantially to completion by oxidative means different from oxidation with N02.

EXAMPLE 2: Synthesis of the antioxidant compounds directed to the mitochondria The chemical synthesis of C3 mitoquinone, C5 mitoquinone and C15 mitoquinone is outlined in Figures 2A-2C and described below. Nuclear magnetic resonance spectra were acquired using a Varian 300 MHz instrument. For 1 H-NMR, the internal standard was tetramethylsilane in CDCl 3. For 3 P-NMR, the internal standard was 85% phosphoric acid. Chemical shifts (d) are in ppm with respect to the standard. The elementary analyzes were done by the Campbell Microanalytical Laboraíory, of the University of Otago. Electrospray mass spectrometry was performed using a Shimadzu LCMS-QP800X liquid chromatography mass spectrometer. Absolute ethanol supply solutions were prepared and stored at -20 ° C in the dark. Mitoquinone C3 (6). The synthetic pathway of mitochonon C3 is shown in Figure 2A. The starting material, 2,3,4,5-tetramethoxytoluene (1) (Lipshutz, BH, Kim, SK, Mollard, P. and Stevens, KL (1998) Tetrahedron 54, 1241-1253), was prepared by reducing 2, 3-dimethoxy-5-methyl-1,4-benzoquinone (C0Q0), to hydroquinol (Carpino, LA, Triolo, SA and Berglund, RA (1989), J. Org. Chem. 54, 3303-3310), followed by methylation to give 1 (Lipshutz, BH, Kim, S.-k "Mollard, P. and Stevens, K. L (1998) Tetrahedron 54, 1241-1253). A solution of 1 (6.35 g, 29.9 mmol) in dry hexane (80 mL) and N, N, NSN'-tetramethylethylenediamine (8.6 mL) was placed under nitrogen with a flame-dried stir bar in a dried Schlenk tube. to the flame. A solution in n-butyllithium hexane (1.6 M, 26.2 mL) was slowly added at room temperature, and the mixture was cooled and stirred at 0 ° C for one hour. After cooling to -78 ° C, dry tetrahydrofuran (THF) was added; 250 mL), and a small aliquot of the reaction mixture was removed, quenched with D20 and examined by 1 H-NMR, to ensure complete metalation. The yellow suspension was then transferred to a second Schlenk tube dried to the flame containing CuCN (0.54 g, 6.03 mmol), under nitrogen and at -78 ° C. The mixture was heated at 0 ° C for 10 minutes and then cooled to -78 ° C and allyl bromide (3.62 mL) was added; the reaction was stirred overnight (19 hours) and allowed to warm to room temperature. The reaction was quenched with 10% aqueous NH 4 Cl (75 mL), and extracted with ether (2 x 200 mL). The combined ether extract was washed with H20 (2 x 150 mL), 10% aqueous NH 4 OH (200 mL) and saturated aqueous NaCl (200 mL). The organic solvent was dried over MgSO4; it was filtered and the solvent was removed by rotary evaporation under vacuum, to give a crude product (7.25 g). Column chromatography on silica gel and elution with 20% ether in hexane, gave pure 1,2,3,4-tetramethoxy-5-methyl-6- (2-propenyl) benzene (2) (Yoshioka, T. , Nishi, T., Kanai, T., Aizawa, Y., Wada, K., Fujita, T. and Horíkoshi, H. (1993), European patent application EP 549366 A1) (6.05 g, 83.5%). 1 H NMR d 5.84-5.98 (1H, m, -CH = C), 4.88-5.03 (2H, m, = CH2), 3.78, 3.80, 3.90, 3.92 (12H, s, OMe), 3.38 (2H, d, J = 7.0Hz, Ar-CH2), 2.14 (3H, s, Ar-Me) ppm. A solution of 2 (8.0 g, 33.05 mmol) in dry THF (45 mL) was added dropwise over 20 minutes, under argon, to a stirred suspension of 9-borabicyclo [3.3.1] nonane in THF (79 mL , 39.67 mmol, 0.5 M) at 25 ° C. The resulting solution was stirred overnight at room temperature and for a further 2 hours at 65 ° C under argon. Then, the mixture was cooled to 0 ° C and 3 M NaOH (53 mL) was added dropwise, followed by 30% aqueous H202 (53 mL). After 30 minutes of stirring at room temperature, the water phase was saturated with NaCl and extracted 3 times with THF. The combined organic fraction was washed with saturated aqueous NaCl, dried (Na2SO4), filtered and evaporated, to give an oily residue (11.5 g) which was purified by column chromatography on silica gel (200 g, packed with ether / hexane 1: 9). An elution with ether / hexane 1: 4 gave pure 3- (2,3,4,5-tetramethoxy-6-methyl-phenyl) -propan-1-o1 (3) as a colorless viscous oil (6.85 g, 80 %). 1 H NMR d 3.91, 3.90, 3.84, 3.79 (12H, s, OMe), 3.56 (2H, t, J = 7.0Hz, -CH2-OH), 2.72 (2H, t, J = 7.0 Hz, Ar-CH2) , 2.17 (3H, s, Ar-Me), 1.74 (2H, quintet, J = 7.0 Hz, -CH2-) ppm. Anal. cale, for C14H2205: C, 62.2; H, 8.2. Found: C, 62.2; H, 8.4%. A solution of 3 (3.88 g, 15 mmol) and triethylamine (3.0 g, 30 mmol, 4.2 mL) in CH2Cl2 (50 mL) was stirred at room temperature for 10 minutes. Methanesulfonyl chloride (1.8 g, 1.20 mL, 15.75 mmol) in CH2Cl2 (50 mL) was added dropwise over 20 minutes, and the reaction mixture was stirred at room temperature for 1 hour. Then, the mixture was diluted with CH2Cl2 (50 mL) and the organic layer was washed with H20 (5 x 100 mL) and 10% aqueous NaHCO3 (100 mL); dried (MgSO4) and filtered, and the solvent was removed under vacuum by rotary evaporation to yield 1- (3-methanesulfonyloxypropyl) -2-methyl-3,4,5,6-tetramethoxybenzene (4), as a liquid ( 4.8 g, 95%). 1 H NMR d 4.27 (2H, t, J = 7.0 Hz, -CH2-0-S02-Me), 3.91, 3.89, 3.82, 3.78 (12H, s, OMe), 3.03 (3H, s, -0-S02- Me), 2.70 (2H, t, J = 7.0 Hz, Ar-CH2-), 2.17 (3H, s, Ar-Me), 1.9 (2H, m, -CH2-) ppm. The crude methanesulfonate 4 (3.30 g, 9.8 mmol) was used directly in the next reaction by mixing with a freshly ground mixture of triphenylphosphine (4.08 g, 15.6 mmol) and Nal (7.78 g, 51.9 mmol) in a Kimax tube, and sealed under argon. Then, the mixture was maintained at 70-74 ° C with magnetic stirring for 3 hours, after which the mixture changed from a thick melted liquid to a crystalline solid. The tube was cooled to room temperature and the residue was stirred with CH2Cl2 (30 mL). Then, the suspension was filtered and the filtrate was evaporated in vacuo. The residue was dissolved in a minimum amount of CH2Cl2 and triturated with an excess of ether (250 mL) to precipitate the white solid. The solid leaked, washed with ether and dried under vacuum, to give pure [3- (2,3,4,5-tetramethoxy-6-methyl-fenll) propyl] -triphenylphosphonium (5) iodide (5.69 g, 90 %). 1 H NMR d 7.82-7.65 (15H, m, Ar-H), 3.88, 3.86, 3.74, 3.73 (12H, s, OMe), 3.76-3.88 (2H, m, CH2-P +), 2.98 (2H, t, J = 7.0 Hz, CH2-Ar), 2.13 (3H, s, Ar-Me), 1.92-1.78 (2H, m, -CH2-) ppm. 31P NMR (121.4 MHz) d 25.32 ppm. Anal. cale, for C32H36l05P: C, 59.8; H, 5.7; P, 4.8; Found: C, 59.8; H, 5.8; P, 4.5%. A solution of the iodide 5 (4.963 g, 7.8 mmol) in CH2Cl2 (80 mL) was stirred with 10% aqueous NaN03 (50 mL) in a separatory funnel for 5 minutes. The organic layer was separated, dried (Na2SO4), filtered and evaporated in vacuo to give the nitrate salt of 5 (4.5 g, 7.8 mmol, 100%), which was dissolved in a mixture of CH3CN and H20 ( 7: 3, 38 mL) and stirred at 0 ° C in an ice bath. Then pyridine-2,6-dicarboxylic acid (6.4 g, 39 mmol) was added, followed by the dropwise addition of a solution of ceric ammonium nitrate (21.0 g, 39 mmol) in CH3CN / H20 (1: 1). , 77 mL) for 5 minutes. The reaction mixture was stirred 20 minutes at 0 ° C and then 10 minutes more at room temperature. The reaction mixture was then poured into H 0 (200 mL) and extracted with CH 2 Cl 2 (200 mL); dried (Na2SO4), filtered and evaporated in vacuo to give a [3- (4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl) propyl nitrate. ] crude triphosphonium (6). The total product was dissolved in CH2Cl2 (100 mL) and stirred 10 minutes with 20% aqueous KBr (50 mL). The organic layer was separated, dried and evaporated in vacuo to give the bromide salt of 6 (4.1 g, 93.6%). 1 H NMR d 7.90-7.65 (15H, m, Ar-H), 4.15-4.05 (2H, m, CH2-P +), 3.96, 3.95, (6H, s, OMe), 2.93 (2H, t, J = 7.0 Hz, CH2-Ar), 2.15 (3H, s, Ar-Me), 1.85-1.70 (2H, m, -CH2-) ppm. 31P NMR d 25.29 ppm. A solution of the bromide of 6 (3.65 g, 6.5 mmol) in CH2Cl2 (75 mL) was stirred with a 10% w / v aqueous solution of sodium methanesulfonate (100 mL) in a separatory funnel for 5 minutes. The CH2Cl2 layer was separated; dried (Na2SO4), filtered and evaporated in vacuo to give the methanesulfonate salt of [3- (4,5-dimethoxy-2-methylene-3,6-dioxo-1,4-cyclohexadiene-1 -yl) -propyltriphenylphosphonium (6) (3.7 g, 98%). 1 H NMR 7.88-7.60 (15H, m, Ar-H), 3.93, 3.92, (6H, s, OMe), 3.90-3.78 (2H, m, CH2-P +), 2.85 (2H, t, J = 7.0 Hz, CH2-Ar), 2.70 (3H, s, OS02CH3), 2.09 (3H, s, Ar-Me), 1.82-1.68 (2H, m, -CH2-) ppm. 3IP NMR (121.4 MHz) d 25.26 ppm. Anal. cale, for C31H33O7PS: C, 64.1; H, 5.7; P, 5.3; S, 5.5. Found: C, 63.8; H, 5.9; S, 5.3; P, 5.2%. Mitoquinone C5 (14). The synthetic route of the mitoquinone C5 is shown in Figure 2B. Dihydropyran (46.83 g, 0.55 mol) was added to 2,3-dimethoxy-5-methyl-1,4-benzohydroquinone (C0Q0) (50 g, 0.275 mol) dissolved in acetic acid (500 mL), and it was stirred at room temperature for 10 minutes. To this solution was added BF3.Et20 (38.57 g, 0.271 mol). The resulting solution was stirred 18 hours at room temperature. After this time, the crude reaction mixture was poured into ice water (500 mL) and extracted with chloroform (1000 mL). The organic extract was washed with brine (500 mL) and dried (MgSO4). The solvent was removed in vacuo to give 2,3-d-methoxy-5-methyl-6- (tetrahydro-pyrn-2-yl) -4- (tetrahydro-pyrn-2). -Ioxy) -phenol (7) crude, as a red oil (115 g), which was used without further purification. A solution of crude 7 (110 g) in a mixture of acetic acid / perchloric acid (97.5: 2.5, 500 mL) was hydrogenated on 5% palladium / carbon (5.42 g) at atmospheric pressure and room temperature until it was finished the incorporation of hydrogen (three days). Then, the reaction mixture was filtered through a pad of Celite and the solid residue was washed with ethanol (500 mL). The combined filtrate was divided into three equal portions and each portion was added to distilled water (1000 mL) and extracted with CH2Cl2 (2 X 200 mL). The combined organic extract was washed with brine (500 mL), saturated sodium bicarbonate solution (500 mL), brine (300 mL) and then dried (MgSO4). The mixture was then filtered and the solvents were removed under vacuum to give crude 4-acetoxy-3- (5-acetoxy-pentyl) -5,6-dimethoxy-2-methyl-phenyl acetate (8), as a red oil (110 g), which was used in the next step without further purification. 1 H NMR d 4.0-4.15 (2H, m, -CH2-0), 3.86 (6H, s, 2x OMe), 2.58 (2H, t, J = 7.0Hz, -CH2-Ar), 2.12 (3H, s, Ar-Me), 2.06 (6H, s, 2x CH3-C = 0), 2.02 (3H, s, CH3-C = 0), 1.35-1.70 (6H , m, -CH2CH2CH2-) ppm. Lithium aluminum hydride (8.0 g, 0.21 mol) was added to dry THF (500 mL) in a 1 L round bottom flask, equipped with a magnetic stirrer, reflux condenser and surrounded by a water bath at room temperature. A solution of crude 8 (74 g) in freshly distilled dry THF (100 mL) was added dropwise to the THF / LiAIH4 mixture over a period of 25-30 minutes. More dry THF (200 mL) was added to facilitate stirring and the reaction was allowed to stir 3 hours at room temperature. Then, the reaction was quenched by the dropwise addition of 3M HCl (20 mL), followed by the slow addition of distilled water (70 mL). Then, the reaction mixture was filtered and the filtrate was washed with brine (2 x 300 mL), dried (MgSO 4), filtered and the solvent removed in vacuo. The green residue remaining in the filter funnel was dissolved in 15% HCl (500 mL) and extracted with CH 2 Cl 2 (1 x 300 mL, 2 x 200 mL). The organic fractions were combined and the product was washed with brine (400 ml), dried (MgSO), filtered and evaporated in vacuo. This extract was combined with the filtrate treatment material to give crude 2- (5-hydroxypentyl) -5,6-dimethoxy-3-methyl-benzene-1,4-diol (9) (68.3 g) as a red oil . This product 9 was purified using column chromatography on silica gel (600 g, packed in 10% ether / CH 2 Cl 2). Elution with 10% ether / CH2Cl2 gave some of 8 unreacted and 2,3-dimethoxy-5-methyl-1,4-benzohydroquinone starting material. Elution with 20% ether / CH2Cl2 gave a mixture of 9 and quinone 10 (14.14 g, 19% from 2,3-dimethoxy-5-methyl-1,4-benzoquinol). Compound 9 was slowly converted to quinone 10 by allowing it to stand in air, and a satisfactory elemental analysis could not be obtained. 1 H NMR d 5.41 (1 H, s, Ar-OH), 5.38 (1 H, s, Ar-OH), 4.88 (6 H, s, 2 x Ar-OMe), 3.65 (2 H, t, J = 6.3 Hz, CH2-OH), 2.61 (2H, t, J = 6.4 Hz, Ar-CH2), 2.14 (3H, s, Ar-Me), 1.42-1.68 (6H, m, 3x-CH2-) ppm. A solution of quinol 9 (7.5 g, 27.7 mmol) in CH2Cl2 (150 mL) was saturated with oxygen gas at atmospheric pressure and a solution of N02 in CH2Cl2 (1 mL, 1.32 M) was added. The reaction was stirred at room temperature under an atmosphere of oxygen for 18 hours, after which a TLC (40% ether / CH2Cl2) showed that the formation of the quinone 2- (5-hydroxypentyl) -5,6-dimethoxy -3-methyl- [1,4] benzoquinone (10) was complete. The solvent was then removed under vacuum to give the product (10) (Yu, C.A. and Yu, L. (1982), Biochemistry 21, 4096-4101) (7.40 g) as a red oil. 1 H NMR d 3.99 (6H, s, 2 x Ar-OMe), 3.65 (2H, t, J = 6.3 Hz, CH2-OH), 2.47 (2H, t, J = 6.3 Hz, Ar-CH2), 2.01 ( 3H, s, Ar-Me), 1.52-1.60 (2H, m, -CH2-), 1.37-1.43 (4H, m, -CH2CH2-) ppm. A solution of 10 (7.40 g, 27.3 mmol) in CH2Cl2 (150 mL) and triethylamine (5.46 g, 5.46 mmol) was prepared and a solution of methanesulfonyl chloride (2.48 g, 30 mL) was added with stirring over 10 minutes. mmol) in CH2Cl2 (50 mL). After stirring an additional 1.5 hours at room temperature, the reaction mixture was washed with distilled water (5 x 100 mL) and a saturated solution of sodium bicarbonate (150 mL), and dried (MgSO4). The mixture was filtered and the solvent was removed in vacuo to give the crude methanol sulfonate (9.03 g) as a red oil. 1H NMR d 4.19 (2H, t, J = 7.5Hz, -CH2-OMs), 3.95 (6H, s, 2x Ar-OMe), 2.98 (3H, s, OS02CH3), 2.44 (2H, t, J = 7.5 Hz, Ar-CHa-), 1.98 (3H, s, Ar-Me), 1.75 (2H, quintet, J = 7.5Hz, -CH2-), 1.38-1.48 (4H, m, -CH2-CH2-) ppm . The methanesulfonate was dissolved in 10% Nal (w / v) in acetone (100 mL) and stirred 44 hours at room temperature. Then, the mixture was concentrated in vacuo and H20 (100 mL) was added to the residue. The mixture was extracted with CH2CI2 (3 x 70 mL) and the combined organic extract was washed with brine, dried (MgSO4), filtered, and the solvent was removed under vacuum, to give 2- (5-iodopentyl) -5Crude 6-dimethoxy-3-methyl- [1,4] benzoquinone (11). This product was purified by column chromatography on silica gel (150 g). Elution with CH2Cl2 and 10% ether / CH2Cl2 gave pure 11 (7.05 g, 69%) as a red oil. 1 H NMR d 3.99 (6H, s, 2 x Ar-OMe), 3.18 (2H, t, J = 6.9 Hz, CH2-I), 2.47 (2H, t, J = 7.2 Hz, Ar-CH2), 2.02 ( 3H, s, Ar-Me), 1.85 (2H, quintet, J = 7.5Hz, -CH2-), 1.38-1.48 (4H, m, -CH2-CH2-) ppm. Anal. cale, for C14H1 I0: C, 44.5; H, 5.1; I, 33.6. Found: C, 44.6; H, 5.1; I, 33.4%. A solution of 11 (1.14 g, 2.87 mmol) in methanol (20 ml) was treated with NaBH 4 (0.16 g, 4.3 mmol) and the mixture became colorless in 1 minute.

After 5 minutes at room temperature, 5% aqueous HCl (100 mL) was added and the solution was extracted with CH2Cl2 (2 x 50 mL). The organic fractions were combined and the product was dried (gS04), filtered and the solvent was removed in vacuo to give 12 (1.15 g, 100%) as a yellow oil sensitive to oxygen, which was used without further delay. 1 H NMR 5.36, 5.31 (2H, s, Ar-OH), 3.89 (6H, s, 2x Ar-OMe), 3.20 (2H, t, J = 7.5Hz, -CH2-I), 2.62 (2H, t , J = 7.5Hz, -CH2-Ar), 2.15 (3H, s, Me), 1.82-1.92 (2H, m, -CH2-), 1.45-1.55 (4H, m, -CH2-CH2-) ppm. A mixture of 12 (1.15 g, 2.87 mmol) and triphenylphosphine (1.2 g, 4.31 mmol) was placed in a Kimax tube with a stir bar. The tube was flushed with argon, sealed tightly and heated and stirred for 14 hours at 70 ° C. A dark solid formed which was dissolved in CH2Cl2 (10 mL) and triturated in ether (200 mL); the white precipitate formed was quickly filtered. The precipitate, which became sticky from exposure to air, was redissolved in CH2Cl2 and evaporated in vacuo to give the crude product, [5- (2,5-dihydroxy-3,4-dimethoxy-6-methyl) iodide. phenyl) -pentyl] -triphenylphosphonium (13) (2.07 g, 115%), as a brown oil. The material is not stable in storage for extended periods and was used as quickly as was practical for subsequent reactions. 1 H NMR d 7.84-7.68 (15H, m, Ar-H), 5.45 (1H, s, Ar-OH), 5.35 (1 H, s, Ar-OH), 3.89 (3H, s, Ar-OMe), 3.87 (3H, s, Ar-OMe), 3.65 (2H, m, -CH2- + PPh3), 2.54 (2H, t, J = 7.0Hz, Ar-CH2), 2.08 (3H, s, Ar-Me) , 1.65-1.75 (2H, m, -CH2-), 1.45-1.55 (4H, m, -CH2CH2-) ppm. 3 P NMR d 25.43 ppm.

A solution of 13 (2.07 g) in CH2Cl2 (50 mL) was saturated with gaseous oxygen and a solution of N02 in CH2Cl2 (0.5 mL, 1.32 M) was added. The reaction was then stirred at room temperature under an atmosphere of oxygen for 18 hours. The solvent was removed in vacuo to give the crude product, [5- (4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl) pentyl] triphenylphosphonium iodide (14). , like a red oil. This residue was redissolved in CH2Cl2 (10 mL) and triturated in ether (200 mL) to give an initial yellow precipitate which froze, forming a red oil in a few minutes. The solvents were decanted and the precipitate was dissolved in CH2Cl2 and the solvent was removed in vacuo to give the product (14) (1866 g) as a red oil. An aliquot (0.880 g) of 14 was purified by column chromatography on silica gel (20 g). The elution with CH2CI2 gave some unidentified purple material. Elution with 5% ethanol / CH2Cl2, gave the pure iodide product 14 (0.606 g) as a red oil. 1 H NMR d 7.84-7.68 (15 H, m, Ar-H) 3.98 (6H, s, 2 x Ar-OMe), 3.65 (2H, m, CH2-P +), 2.40 (2H, t, J = 7.5 Hz , Ar-CH2), 2.00 (3H, s, Ar-Me), 1.71 (4H, m, -CHjr), 1.43 (2H, m, -CH2-) ppm. 31P NMR (121.4 MHz) d 25.47 ppm. Anal, cale, for C32H36IO4P: C, 59.8; H, 5.7; I, 19.8; P, 4.8; Found: C, 60.0; H, 5.3; I, 19.7; P, 4.7%. Mitoquinone C15 (16). The synthetic route of the mitochonone C15 is shown in Figure 2C. A solution of K2S208 (0.450 g, 1.66 mmol) in H20 (25 mL) was added dropwise over 2.5 hours to a stirred suspension of AgN0 (0.262 g, 1.54 mmol), 16-hydroxyhexadecanoic acid (0.408 g, 1.50 mmol) , and 2,3-dimethoxy-5-methyl-1,4-benzoquinone (0.271 g, 1.49 mmol) in H20: CH3CN (1: 1, 36 mL), maintained at 75 ° C. After stirring 30 minutes, the mixture was cooled and extracted with ether (4 x 30 mL). The combined organic phase was washed with H20 (2 x 100 mL), NaHCO3 (1 M, 2 x 50 mL) and saturated NaCl solution (2 x 50 mL). The organic phase was dried (Na2SO4), filtered and concentrated in vacuo to give a red oil (0.444 g). Column chromatography of the crude oil (silica gel, 15 g) and elution with mixtures of CH 2 Cl 2 and ether (0%, 5%, 20%), gave 2- (15-hydroxypentadecyl) -5,6-dimethoxy- 3-methyl- [1,4] benzoquinone (15) (0.192 g, 33%) as a red oil. 1H NMR d 3.99, 3.98 (6H, s, OMe), 3.64 (2H, t, J = 6.5Hz, -CH2OH), 2.45 (2H, t, J = 7.5Hz, -CH2-ring), 1.4-1.2 ( 26H, m, - (CH2) 13-). Anal. cale, for C24H4o05: C, 70.6; H, 9.9. Found: C, 70.5; H, 9.8%. A mixture of triphenylphosphine (0.066 g, 0.25 mmol), Ph3PHBr (0.086 g, 0.25 mmol) and 15 (0.101 g, 0.25 mmol) was stirred under argon in a sealed Kimax tube at 70 ° C for 24 hours, after which had turned into a viscous red oil. The residue was dissolved in a minimum amount of CH2Cl2 (0.5 mL) and emptied into ether (10 mL) to produce a red oily precipitate. The solvents were then decanted and the residue was dissolved in CH3OH (0.5 mL) and diluted in H20 (10 mL) containing 48% HBr (1 drop). A red precipitate formed and after the precipitate settled, the supernatant was emptied and the residue was washed with H20 (5 mL). Then, the residue was dissolved in ethanol (5 mL) and the solvent was removed in vacuo. The residue was redissolved in CH2CI2 (0.5 mL), diluted with ether (5 mL) and the solvent decanted; the residue was placed in a vacuum system (0.1 mbar) for 24 hours to give [15- (4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl) bromide) pentadecyl] triphenylphosphonium (16) (0.111 g, 61%), as a yellow foam that turned into a red oil upon contact with air. 1 H NMR (299 MHz) d 7.6-8.0 (15H, m, Ar-H), 3.89 (6H, s, OMe), 3.9 (2H, m, -CH2-P), 2.6 (2H, m, -CH2- ring), 1.7-1.1 (26H, m, - (CH2) 13-) ppm. 31P NMR (121.4 MHz) d 25.71 ppm. Electrospray mass spectrometry: found (M +) 653, calculated for C42H54? 4P + 653. The analytical combustion results were not satisfactory due to inconsistent levels of solvent inclusion.

Example 3: Properties of antioxidant compounds directed to the mitochondria The present invention recognizes that to be suitable in a wide variety of applications, for example in the formulation of dosage forms such as tablets, it is advantageous to be able to form a crystalline or solid form of the mitochondria-targeting compound. Similarly, it is considered, without wishing to be bound by any theory, that the antioxidant functionality of the compounds of the present invention is determined, at least in part, by their physicochemical properties. Table 1 shows the partition coefficients of a variety of antioxidant compounds. The partition coefficients 1-octanol / PBS were determined by adding 400 nmol of the compound to 2 ml of 1-octanol saturated with PBS, and mixing for 30 minutes at 37 ° C with 2 ml of PBS saturated with 1-octanol. The concentrations of the compound in the two phases were measured by UV absorption at 268 nm, and were quantified from standard curves of the compound in PBS saturated with 1-octanol, or 1-octanol saturated with PBS (Kelso, GF, Porteous, CM, Coulter, CV, Hughes, G., Porteus, WK, Ledgerwood, EC, Smith, RAJ, and Murphy, MP, 2001, J Biol Chem 276, 4588, Smith, RAJ, Porteous, CM, Coulter, CV, and Murphy, MP 1999 Eur J Biochem 263, 709). Compound solutions were prepared in absolute ethanol and stored at -20 ° C in the dark. The [3H] TPMP was from American Radiolabelled Chemicals Inc, (Missouri, E.U.). Particularly notable is the low partition coefficient of the compounds with few carbon atoms of the bridge of the antioxidant portion with the phosphonium. For example, a compound within the present invention, referred to herein as C3-mitoquinone, having a 3-carbon bridge, has a partition coefficient approximately 50 times lower than that observed for the related compound, mitochonon C10 (Table 1).

Table 1: Partitioning coefficients of antioxidants and related compounds The data at "c are partition coefficients of 1-octanol / phosphate buffer, determined at 25 ° C or 37 ° C as described above, or octanol / water partition coefficients calculated using Advanced Chemistry Development (ACD) Solaris V4.67 software, as described in Jauslin, ML, Wirth, T., Meier, T._ and Schoumacher, F., 2002, Hum Mol Genet 11, 3055. to Kelso, GF, Porteus, CM, Coulter, CV, Hughes, G., Porteus, WK, Ledgerwood, E. C, Smith, RAJ, and Murphy, MP, 2001, J Biol Chem 276, 4588. b Smith, RAJ, Porteous, CM, Coulter, CV, and Murphy , MP 1999, Eur J Biochem 263, 709. 0 Smith, RAJ, Porteous, CM, Gane, AM, and Murphy, MP, 2003, Proc Nat Acad Sci 100, 9, 5407. From their partition coefficients in 1-octanol / PBS is evident that mitochonon C3, mitoquinone C5, mitoquinone C10 and mitoquinone C15 encompass a wide scale of hydrophobicity.The one of mitoquinone C3 is similar to the simple cation TPMP relatively luble in water, while that of mitochonon C15 indicates that it has very low solubility in water. It is reported that alkyltrifenifosphonium cations, such as mitoquinone, are adsorbed on phospholipid bilayers with the cation in the carboxylic acid groups, while the hldrophobic alkyl group penetrates the hydrophobic core of the membrane. It is considered that the larger the methylene bridge the antioxidant ubiquinol will penetrate deeper into the hydrophobic core of the membrane. The present authors believe that the maximum penetration in a leaflet of the membrane will occur for these compounds as illustrated in Figures 3A-3F, which show the variants of mitoquinone aligned with a typical phospholipid. These models indicate that the ubiquinol portion of the C3 mitoquinone only penetrates near the membrane surface, while that of C10 and C10 mitochonone penetrates near the nucleus of the phospholipid layer.

The present authors synthesized a series of antioxidant compounds with a range of hydrophobicity and penetration depths in the phospholipid layer.

EXAMPLE 4: Mitochondrial incorporation of compounds directed to the mitochondria To demonstrate that mitochondrial targeting is effective, incorporation into the mitochondria was determined in response to the membrane potential of the exemplary anti-oxidant compounds mitoquinone C3, mitoquinone C5, mitochonone C10, and mitoquinone C15. To measure the incorporation of antioxidant compounds into energized mitochondria, an ion-selective electrode was constructed (Smith, RA, Kelso, GF, James, AM and Murphy, MP (2004), Met. Enzymol. 382, 45-67; , GP, Tipton, KF and Murphy, MP (1992), Biochem J. 288, 439-443, Kamo, N., Muratsugu, M., Hongoh, R. and Kobatake, Y. (1979) J. Membr. Biol. 49, 104-121). The electrode and an Ag / AgCI reference electrode were inserted through the air-tight Perspex lid of a 3 ml incubation chamber, shaken and thermally adjusted at 30 ° C, provided with an injection orifice for the addition of substrates. To measure the incorporation of the antioxidant compound, mitochondria from rat liver (1 mg protein / ml) were incubated at 30 ° C in KCl medium (120 mM KCl, 10 mM HEPES, pH 7.2, 1 mM EGTA) and nigericin was added. (1 μg / ml) and rotenone (8 μg / ml). Succinate (10 mM) and FCCP (500 nM) were added where indicated. The output of the ion selective electrode was passed to a PowerLab Data data acquisition system by means of a front-end pH amplifier and analyzed using the Chart software, all from ADInstuments. Rat liver mitochondria were prepared by homogenization, followed by differential centrifugation in ice-cooled buffer containing 250 mM sucrose, 5 mM Tris-HCl, 1 mM EGTA, pH 7.4 (Chappell, JB and Hansford, RG (1972 ) in: "Subcellular components: Preparation and fractionation", pp. 77-91 (Birnie, GD, ed.) Butterworths, London). The protein concentration was determined by means of the biuret test using BSA as a standard (Gornall, A.G., Bardawill, C.J., and David, M.M. (1949), J. Biol. Chem. 177, 751-766). The mitochondrial membrane potential was measured by adding 500 nM of TPMP supplemented with 50 nCi of [3 H] TPMP to mitochondria suspended in KCl medium (120 mM KCl, 10 mM HEPES, pH 7.2, 1 mM EGTA) at 25 ° C (Brand, MD (1995) in: "Bioenergetics -a practical approach", pp. 39-62 (Brown, GC and Cooper, CE., Eds.) IRL, Oxford). After incubation, the mitochondria were pelleted by centrifugation and the amount of [3H] TPMP in the supernatant and in the pellets was quantified by scintillation counting, and the membrane potential was calculated assuming a mitochondrial volume of 0.5 μl / mg of mitochondrial protein and a correction for TPMP binding of 0.4 (Brown GC and Brand, MD (1985), Biochem. J. 225, 399-405).

The present authors constructed elective electrodes to measure their constant state concentrations (Smith, RA, Kelso, GF, James, AM and Murphy, MP (2004) Meth, Enzymol, 382, 45-67, Davey, GP, Tlpton, KF and Murphy, MP (1992) Biochem J. 288, 439-443; Kamo, N., Muratsugu, M., Hongoh, R. And Kobatake, Y. (1979), J. Membr. Biol 49, 105-121) . The response of these electrodes to simple triphenylphosphonium cations such as TPMP is Nernstiana, with a linear response of electrode voltage at log.o [cation concentration] and a slope of ~ 60mV at 30 ° C (Davey, GP, Tipton , KF and Murphy, MP (1992) Biochem, J. 288, 439-443; Kamo, N., Muratsugu, M., Hongoh, R. and Kobatake, Y. (1979) J. Membr. Biol. 49, 105 -121). The more hydrophilic compound, mitoquinone C3, also gave a Nernstiana electrode response with a slope close to 60 mV at concentrations above 10 μM. This is illustrated in FIG. 4A, on the right hand side, by the logarithmic electrode response to the sequential additions of 1 pM of C3 mltoquinone in the absence of mitochondria. For C5 mitoquinone, C10 mitoquinone and C15 mitoquinone, the electrode also responded rapidly and stably to sequential additions in the absence of mitochondria (Figures 4B, 4C and 4D, respectively, panels on the right side). However, in these cases the electrode responses were not Nernstianas; the authors consider that this was due to the greater hydrophobicity of these compounds. Even so, for the four antioxidant compounds, the ion selective electrode allowed the measurement of the free concentrations of the compounds and therefore their incorporation into the mitochondria in real time. To measure the incorporation of the antioxidant compound, mitochondria were added to the electrode chamber in the presence of rotenone to prevent the formation of a membrane potential (left side of Figures 4A-4D). Then, five sequential additions of 1 μM of antioxidant compound were made to calibrate the electrode response, followed by the succinate respiratory substrate to generate a membrane potential. Mitochondrial energlization leads to the rapid incorporation of all variants of antioxidant compounds by the mitochondria, and the subsequent addition of the FCCP uncoupler suppressed the membrane potential and led to its rapid release from the mitochondria (Figures 4A-4D, left side ). These experiments clearly show the membrane-dependent uptake of mitochonon C3, mitoquinone C5, and mitochonon C10. Although mitochondrial C15 was also taken by the mitochondrion by induction of a membrane potential, the electrode response to mitochondrial C15 in the presence of mitochondria was weaker, more noisy and more susceptible to drift. This contrasts with the electrode response to mitochondrial C15 in the absence of mitochondria (see panels on the right side), and is due to its low free concentrations in the presence of mitochondria. The magnitude of the antioxidant compound binding to de-energize the mitochondria was then determined (Figures 4A-4D, right side). For these experiments the antioxidant compound variants were first added to the electrode chamber and then the mitochondria were added in the presence of rotenone to prevent the formation of a membrane potential. The reduction of the concentration of antioxidant compound when adding the mitochondria is due to the union of the antioxidant compound with the denergized mitochondria. The subsequent addition of succinate to generate a membrane potential indicates the incorporation of the compounds dependent on the membrane potential, which is then reversed with the addition of FCCP to suppress the membrane potential. The concentration of free C3 mitochonon was not affected by the addition of mitochondria, indicating that negligible amounts of mitochondrone C3 bound to the de-energized mitochondria (Figure 4A right side). The FCCP-sensitive incorporation of mitochondrone C3 by energization with succinate was approximately 3.7 nmol of C3 mitoquinone / mg protein, corresponding to an accumulation ratio of -2x103. This is consistent with what was expected from the Nernst equation and a mitochondrial membrane potential of approximately 180 mV, allowing corrections for the intramitochondrial junction. For mitochondon C5 there was some binding of the compound to the de-energized mitochondria (~ 0.6 nmol / mg protein); however, this was negligible in comparison to its subsequent incorporation by succinate energization, of approximately 2.8 nmol of C5 mitochonon / mg protein, which corresponds to an accumulation ratio of approximately 1.4x103 (Figure 4B, right side).

For C10 mitochonone there was significant binding to the de-energized mitochondria of approximately 2.6 nmol of C10 mitochonone, and this was followed by additional incorporation of approximately 1 nmol / mg protein with the addition of succinate (Figure 4C, right side). Almost all free C15 mitochonone bound to the de-energized mitochondria, but there was some additional incorporation by energization with succinate. The incorporation of mitochondrial C15 dependent on the membrane potential was clear in the left panel of Figure 4D, where the electrode response was very sensitive to allow measurement of the small amount of free C15 mitoquinone when the electrode was calibrated in the presence of mitochondria. In contrast, it is difficult to observe the incorporation of mitochondrial C15 on the right side of Figure 4D, where the electrode response was much less sensitive to allow the measurement of C15 mitoquinone in the absence of mitochondria. These experiments show that the length of the methylene bridges of the antioxidant compounds determines, at least in part, their magnitude of adsorption to the mitochondrial membranes (right side of Figures 4A-4D). The adsorption varies from negligible for C3 mitoquinone to almost complete binding for the C15 mltoquinone. By adding C15 mitoqulnone to the de-energized mitochondria, essentially all the compound binds, distributed through both membrane surfaces, internal and external. When a membrane potential is induced, the present authors consider that there will be a significant redistribution of the compound to the surface facing the matrix of the inner membrane, from the outer surface of the inner membrane, and from the outer membrane. In summary, all variants of the antioxidant compound are incorporated into the mitochondria driven by the membrane potential, and the larger the methylene bridge, the greater its adsorption in the phospholipid bilayers.

EXAMPLE 5: Antioxidant efficacy of the compounds directed to the mitochondria.

The compounds of the invention are also very effective against oxidative stress. To measure antioxidant efficacy, the ability of antioxidant compounds to prevent peroxidation of lipid in the mitochondria was measured from the accumulation of TBARS in mitochondria exposed to ferrous ion and hydrogen peroxide (Figures 5A-5D). To quantify lipid peroxidation, the TBARS test was used. Rat liver mitochondria (2 mg protein / ml) was incubated in 0.8 ml of medium containing 100 mM KCl, 10 mM Tris-HCl, pH 7.6, at 37 ° C, supplemented with 10 mM succinate and rotenone 8 mg / ml, or an ATP regeneration system of 2.5 mM ATP, 1 mM phosphoenolpyruvate and 4 U / ml pyruvate kinase. Then, the mitochondria were exposed to oxidative stress by adding 50 mM FeCl2 / 300 mM H202 for 15 minutes at 37 ° C. After incubation, 64 ml of 2% butylated hydroxytoluene (w / v) in ethanol was added, followed by 200 ml of 35% (v / v) HCl04 and 200 ml of 1% thiobarbituric acid (w / v). ). Then, the samples were incubated for 15 minutes at 100 ° C, centrifuged (5 min at 12,000 x g), and the supernatant was transferred to a glass tube. After the addition of 3 ml of water and 3 ml of 1-butanol, the samples were mixed with vortex and the two phases were allowed to separate. Aliquots of 200 ml of the organic layer were then analyzed in a fluorometric plate reader (? Ex = 515 nm;? Em = 553 nm), to determine the reactive species of thiobarbituric acid (TBARS) and compared with a standard curve of malondialdehyde (MDA), prepared from 0.01-5 mM of 1,1,3,3-tetraethoxypropane (Kelso, GF, Porteous CM, Coulter, CV, Hughes, G., Porteous, WK, Ledgerwood, EC, Smuth RAJ and Murphy , MP (2001) J. Biol. Chem. 276, 4588-4596). For mitochondria energized with succinate, the background level of TBARS was negligible but increased to about 3.75 nmol of MDA / rpg protein by exposure to oxidative stress (Figure 5A; black bars). High concentrations (5 μM) of any of the antioxidant compounds greatly prevented the accumulation of TBARS, while the simple TPMP cation did not. This confirms that it was the ubiquinol side chain of the mitochondrial antioxidant compounds, responsible for the antioxidant action, and not any specific interaction of the cation with the mitochondria. In these experiments, succinate maintains a membrane potential to manage the accumulation of cations in the mitochondria, and also recycles the ubiquinone form of mitochondrial antioxidant compounds to the antioxidant active form of ubiquinol (Kelso, GF, Porteous, CM, Coulter ,, CV, Hughes, G., Porteous WK, Ledgerwood, EC, Smith, RAJ and Murphy, MP (2001) J. Biol. Chem. 276 4588-4596). To see if reduction with the respiratory chain was necessary for the antioxidant efficacy of the mitochondon antioxidant compounds, the present authors incubated mitochondria in the presence of ATP and an ATP regeneration system. The hydrolysis of ATP and the reversal of mitochondrial ATP synthase produced an extensive proton pump that formed a membrane potential similar to that generated by succinate (Figure 5B). This led to the same incorporation of mitochondrial antioxidant compound as mitochondria energized by succinate, but now mitochondon amphotericin compounds are no longer recycled to their active forms of ubiquinol by the respiratory chain. Mitoquinone antioxidant compounds were ineffective in preventing lipid peroxidation of the mitochondria energized by ATP hydrolysis (Figure 5A, white bars), compared to the dramatic protection observed in the mitochondria energized by succinate (Figure 5B, black bars). Therefore, the reduction of mitochondrial antioxidant compounds by the respiratory chain, as well as the accumulation by the mitochondrial membrane potential, are necessary for the antioxidant efficacy of the mitochondrial antioxidant compounds.

Lower degrees of lipid peroxidation were observed in mitochondria control samples energized with succinate, compared to those energized with ATP (Figure 5A). This is due to the protective antioxidant effect of the endogenous mitochondrial reserve of coenzyme Q, which is reduced by the presence of succinate, but oxidized in the presence of ATP (James, AM, Smith, RA and Murphy, MP (2004) Arch. Biochem. Biophys 423, 47-56, Emster, L, Forsmark, P. and Nordenbrand, K. (1992) Biofactors 3, 241-8). In summary, all mitochondrial antioxidant compounds require activation by the respiratory chain to be effective antioxidants. For Figure 5A a single concentration of 5 pM was used for all the mitochondone antioxidant compounds. To compare their relative antioxidant efficiencies the compounds were titrated in the presence of succinate: a typical titration is shown in Figure 5C. This experiment suggests that the antioxidant efficacy of these compounds correlates with the length of the methylene bridge. To quantify this, Cl50 values were calculated for the prevention of lipid peroxidation with the four exemplary mitochondrial antioxidant compounds (Figure 4D). These experiments confirmed that the order of antioxidant efficacy was: mitoquinone C15 > C10 mitoquinone > C5 mitoquinone > Mitoquinone C3. All the mitochondrial antioxidant compounds accumulated in the mitochondria driven by the mitochondrial membrane potential. For the more hydrophobic compound, mitochondrial C15, this effect was masked mainly by extensive binding to the phospholipid bilayers. All the compounds were effective antioxidants and for a persistent antioxidant activity for 15 minutes all required the action of the respiratory chain to recycle the antioxidant compound mitoquinone to its antioxidant form acfiva, after having detoxified the lipid peroxidation intermediates.

Example 6: Effect of antioxidant compounds directed to mitochondria on cardiac hemodynamics and mitochondrial function The effect of the administration of antioxidant compounds directed to mitochondria, in particular C10 mitoquinone and C3 mitoquinone, on cardiac function, was determined using Langendorff's isolated heart perfusion model. Rats were assigned to the following four administration groups: Control (placebo), TPMP (methyltriphenylphosphonium), Mitoquinone-C10 and Mitoquinone-C3. After the treatment period, the rats were sacrificed humanely and the isolated hearts were connected to the isolated perfusion system of Langendorf. This system uses retroperfusion through the aorta to maintain the heart while measuring cardiac function. The left ventricular pressure was measured with a left ventricular balloon. The coronary flow was also measured. Figure 6 shows the coronary flow at 10 mm Hg of the left ventricular pressure for each treatment group. Coronary flow was measured before ischemia and again at zero minutes, 20 minutes, 40 minutes and 60 minutes after the induction of ischemia. A one-way ANOVA was done with a post hoc Bonferroni test. Significance against preischemic control: * P < 0.05; ** P < 0.01; *** P < 0.001. Significance against the respective time control: t P < 0.05; tt P < 0.01; ttt P < 0.001. The results show that treatment with C10 mitoquinone significantly decreases coronary flow reduction induced by ischemia. Mitoquinone C3 has a minor but still significant effect at later time points. The absence of any effect with the administration of TPMP indicates that it is the antioxidant portion of the C10 mitoquinone and the C3 mitoquinone, and not the triphenylphosphonium cation, responsible for the effects observed with the antioxidant compounds directed to the mitochondria. Figure 7 depicts the effects of treatment on left ventricular diastolic pressure at 10 mm Hg. The left ventricular diastolic pressure was measured before inducing ischemia and again at zero minutes, 20 minutes, 40 minutes and 60 minutes after the induction of ischemia. The statistical analysis was a serial ANOVA with a post hoc Dunns test. Significance against preischemic control: * P <; 0.05; t represents P < 0.05 against posischemic control at 60 min. The results show that treatment with mitochondrone C10 results in a statistically significant increase in left ventricular diastolic pressure against untreated rats, decreasing the reduction in left ventricular diastolic pressure induced by ischemia. The absence of any effect with the administration of TPMP indicates that it is the antioxidant portion of mitochondrone C10, and not the cation of triphenylphosphonium, that is responsible for the effects observed with the antioxidant compounds directed to the mitochondria. The effect of the administration of C10 mitoquinone and mitoquinone C3 on the heart rate was then determined. Figure 8 represents the heart rate for each treatment group before ischemia, and at zero minutes, 20 minutes, 40 minutes and 60 minutes after the induction of ischemia. The results shown are a one-way ANOVA, followed by a post hoc bonferroni test. *** represents P < 0.001 against ischemic control, tt represents P < 0.05 against the respective posischemic control. The results show that the treatment with mitochondone C10 significantly decreases the reduction in heart rate induced by ischemia, in comparison with the control rats. The absence of effects with the administration of TPMP indicates that it is the antioxidant portion of mitochondrone C10, and not the cation of triphenylphosphonium, that is responsible for the effects observed with the antioxidant compounds directed to the mitochondria. Cardiac function was further examined by determining the effect of the administration of antioxidant compounds directed to the mitochondria on the frequency of contraction and relaxation of the heart. Figure 9A represents the frequency of contraction in each of the four treatment groups before Ischemia, and at zero minutes, 20 minutes, 40 minutes and 60 minutes after the induction of ischemia. Figure 9B represents the relaxation frequency in each of the four treatment groups before ischemia, and at zero minutes, 20 minutes, 40 minutes and 60 minutes after induction of ischemia. In each case an ANOVA was performed in series with a post hoc Dunns test. * represents significance with P < 0.05 against the control of preischemia. t represents significance with P < 0.05 against the respective posischemic time controls, tt represents significance with P < 0.01 against the respective posischemic fime control. The results show that the administration of mitochondrone C10 has a statistically significant effect, decreasing the reduction of the frequency of contraction and relaxation of the left ventricle induced by ischemia, in comparison with the control rats. The above data clearly show the beneficial effect of the administration of antioxidant compounds directed to the mitochondria on cardiac function. To determine if the observed effects on cardiac function were due to the effect of the antioxidant compound directed to the mitochondria on mitochondrial function, preischemic and posischemic mitochondrial activity was determined for each treatment group. Figure 10A represents the respiratory function linked with NAD + of the pre- and posischemic mitochondria for each of the four treatment groups. Figure 10B shows the respiratory function linked to pre- and posischemic FAD for each of the four treatment groups. *** represents significance with P < 0.001 against preischemic control. ttt represents significance with P < 0.001 against posischemic control. These data show that mitochondrial C10 has a statistically significant beneficial effect on mitochondrial respiratory function after ischemia, in comparison with control rats. These results support the conclusion that the effects of the administration of antioxidant compounds directed to the mitochondria on cardiac function are due to a protective effect on mitochondrial function.

Example 7: Stability of C10 mitochonone complexes with ß-cyclodextrin In preformulation studies it was found that mitochonon C10 in its salt form bromide degrades over time in the solid state when stored at 25 ° C and 50% RH, and at 40 ° C and 75% RH. The objective of the present study was to establish if the stability of the mitochondrion C10 in the solid state could be improved by placing it in complex with ß-cyclodextrin. Mitoquinone C10, lot No.6, and idebenone, were provided by Industrial Research Limited (New Zealand). The β-cyclodextrin (batch No. 70P225) was purchased from ISP Technologies Inc. In addition, NaCl, NaH PU and methanol (HPLC) were purchased from BDH. Study of the stability of mitochondrion C10 in the solid state Specimens of mitochonon C10 were accurately weighed (approximately 5 mg) in clear containers and exposed to 25 ° C and 50% RH, 40 ° C and 75% RH and 4 ° C on silica. The containers were removed after 1, 2, 4, 8, 16, 32 and 64 days and their content of C10 mitoquinone was analyzed by means of a validated HPLC method, using as control mitochonone C10 stored at -20 ° C on silica . Preparation of complexes of mitochonone C10: 6-cyclodextrin Using lot 6 of mitoquinone C10, three complexes with different molar proportions were prepared (mitochondone bromide C10 : β-cyclodextrin, 1: 1, 1: 2 and 1: 4). Preparation of a solution of 3-cyclodextrin in aqua Exactly ß-cyclodextrin was weighed (1137 g equivalent to 1.0361 g after correction for moisture content), and dissolved by sonication for 10 min in double distilled water. The volume was brought to 100 ml with water. Preparation of the C10 mitoquinone complex: β-cyclodextrin (1: 1 molar ratio) An ethanolic solution of C10 mitochonone bromide (90 mg equivalent to 59.95 mg of C10 mitoquinone) was evaporated under nitrogen in a hot plate maintained at 40-50 ° C for 8 min. Β-Cyclodextrin solution (10 ml) and double distilled water (30 ml) were added to the flask, and then sonicated for 40 min. Preparation of the C10 mitoquinone complex: β-cyclodextrin (1: 2 molar ratio) An ethanolic solution of C10 mitochonone bromide (89.8 mg equivalent to 59.82 mg of C10 mitochonone), was evaporated under nitrogen in a hot plate maintained at 37-45 ° C for 10 min, followed by 3 min at 50 ° C. Β-Cyclodextrin solution (20 ml) and double distilled water (20 ml) were added to the flask, and then sonicated for 30 min. Preparation of the C10 mitoquinone complex: β-cyclodextrin (1: 4 molar ratio) An ethanolic solution of C10 mitochonone bromide (90 mg equivalent to 59.95 mg of C10 mitoquinone) was evaporated under nitrogen in a hot plate maintained at 37-50 ° C for 12 min. Β-cyclodextrin solution (40 ml) was added to the flask, and then subjected to sonication for min. The above solutions were frozen keeping them at -18 ° C overnight. The frozen solutions were freeze-dried for 2 days using the LABCONO apparatus. The lyophilized compounds were stored at -20 ° C. Differential Scanning Calorimetry of C10-Mitoquinone Complexes: β-cyclodextrin Freeze-dried Differential Scanning Calorimetry (DSC) was performed on the three freeze-dried complexes using a Perkin Elmer PYRIS-1 Differential Scanning Calorimeter. A sample of C10 mitoquinone was prepared by evaporating an ethanolic solution under nitrogen gas at 35-50 ° C for 10 min. Aluminum crucibles were used (No. 0219-0041, provided by Perkin-Elmer). The analysis was done under a nitrogen purge. Empty crucibles were used to establish the baseline. The sweep temperature scale was 50-160 ° C with an initial retention at 50 ° C for 1 min, followed by an increase of 10 ° C / min up to 160 ° C. HPLC test An HPLC method for C10 mitoquinone using methanol and 0.01 M sodium dihydrogen phosphate (85:15) was developed as the mobile phase, at a flow rate of 1 ml / min and using UV-VIS detection at 265 nm. The internal standard was idebenone. The column was Prodigy ODS3100A (Phenomenex) with a particle size of 5 μ. Subsequently, this method was modified after the arrival of a new column. The mobile phase used in the modified method was methanol and 0.01 M sodium diacid phosphate (80:20). This method was validated. The interference by β-cyclodextrin in the HPLC method was verified before analyzing the complexes of mitochonone C10: β-cyclodextrin. It was observed that β-cyclodextrin does not affect the HPLC test of C10 mitoquinone.

Stability study of the complexes of mitochondone C10: β-cyclodextrin As were three complexes of mitochonone C10 with β-cyclodextrin, the amount of mitochonone C10 in samples of 5 mg of the different complexes, was different. To expose equal amounts of C10 mitoquinone in the three complexes, different weights of the complexes were taken: 4 mg of the 1: 1 complex containing 1473 mg of C10 mitochonone; 6.5 mg of a 1: 2 complex containing 1,469 mg of C10 mitoquinone; and 11.5 mg of complex containing 1467 mg of C10 mitochonone, and were used in the stability study according to the standard operating procedure. Aliquots of HPLC water (1.5 ml) were added to each sample vessel to completely dissolve the C10 mitochonone: β-cyclodextrin complexes. Aliquots of these solutions (50 μl) were diluted to 1 ml with water. Aliquots of these diluted solutions of the mitochonone C10: β-cyclodextrin complexes (100 μl) were vortexed with 200 μl of a 10 μg / ml internal standard solution in methanol. The samples were centrifuged for 10 min at 10,000 rpm and 50 μl of the supernatants were injected into the HPLC system. A standard curve was prepared using C10 mitochonone solutions in the concentration range of 2.5 to 120 μg / ml containing 5 mg / ml solutions of β-cyclodextrin. All the compounds had a faint orange-yellow color and a very spongy appearance. The color was not uniform and was more concentrated towards the bottom of the freeze drying flasks.

The results of the DSC are given as follows: Mitochonone-C10: When a pure sample of C10 mitoquinone was analyzed, peaks above 120 ° C were observed. With a sample of C10 mitoquinone, two prominent peaks between 130 ° C and 140 ° C were observed. When another sample was analyzed, no such prominent peaks were observed but small peaks above 120 ° C were observed. After the analysis, the crucibles were cut and the samples examined. The samples were dark green to black in both cases. β-Cyclodextrin: There was a broad peak between 70 ° C and 85 ° C. Mitoquinone complex C10: β-cyclodextrin (1: 1): No significant peaks were observed. After the analysis the crucible was cut and examined. The color of the sample had undergone a slight change to light brown (it is not a significant change). Mitoquinone complex C10: β-cyclodextrin (1: 2): No significant peaks were observed. After the analysis no color change was observed in the sample. C10 mltoquinone complex: β-cyclodextrin (1: 4): No significant peaks were observed but a very small exothermic peak was observed at 120 ° C. After the analysis no color change was observed in the sample. The appearance of the peaks in the sample of pure C10 mitoquinone indicates that changes of the compound occur with temperature. However, as there were many peaks and also color changes in the sample, these could have arisen due to degradation. When a second sample of mitochonone C10 was analyzed, it gave a thermogram different from the first sample. In the case of complexes, there were no significant peaks or color changes. The results of the study of the stability of mitochonon Pure C10 (batch No. 3) in the solid state are given in Table 2 and Figure 11.

Table 2: Stability of C10 mitoquinone (batch No. 3) in solid state Stability of the C10 mitoquinone (lot No. 3) in the solid state in the absence of light at 40 ° C and 75% RH; 25 ° C and 50% RH; and 5 ° C on blue silica gel; the data are averages of two values expressed as a percentage of the original content.

Due to significant instability at 25 ° C and 50% RH compared to 40 ° C and 75% RH, the stability study was repeated at 25 ° C and 50% RH with C10 mitoquinone, batch No. 4. The second stability study was carried out in both transparent and amber containers, and the results are given in table 3 and figure 12.

Table 3: Stability of C10 mitoquinone (batch No. 4) in solid state Stability of C10 mitoquinone (lot No. 4) in the solid state in the absence of light at 25 ° C and 50% RH; the data are means of three values expressed as a percentage of the initial content.

Both batches (Nos. 3 and 4) of C10 mitochonone provided by the Chemistry Department showed a sudden drop in content after 16 days. However, in batch No. 4 the degradation was not as great after 32 to 64 days compared to batch No. 3. It was also observed that the stability of the C10 mitoquinone is independent of whether the containers are transparent or amber . The C10 mitoquinone provided by IRL was used for the preparation of the C10 mitoquinone: β-cyclodextrin complexes. The C10 mltoquinone provided by IRL was a reddish yellow syrup in ethyl alcohol. The stability of the complexes of mitochondone C10: β-cyclodextrin is given in table 4 and figures 13, 14 and 15. Due to the small amounts available for study of the complexes of mitochonon C10: β-cyclodextrin, there were no results for day 1 and day 4.

Table 4: Stability of C10 mitoquinone complexes: β-cyclodextrin in solid state Stability of the C10 mitoquinone complex: β-cyclodextrin in the solid state in the absence of light at 40 ° C and 75% RH; 25 ° C and 50% RH; and 5 ° C on blue silica gel; the data are averages of two values expressed as a percentage.

The results show that C10 mitochonon can efficiently complex with β-cyclodextrin and can be complexed with β-cyclodextrin. The results show that the C10 mitoquinone in the β-cyclodextrin complexes 1: 1 and 1: 2 are stable under various conditions. The results also show that the stability of the C10 mitoquinone in the 1: 4 complex was reduced with respect to the stability of the C10 mitoquinone in the β-cyclodextrin complexes 1: 1 and 1: 2.

Example 8: Stability studies of mitochonon mesylate C10 Stability of the C10 mitoquinone mesylate in solution The stability of the mitochonone mesylate C10 in solution was determined in five solvents: water, 0.01 M HCl, 0.01 M NaOH, IPB (pH 7.4) and 50% MeOH, at two temperatures: 25 ° C and 40 ° C, under two atmospheric conditions: air and nitrogen, for 125 days, according to the applicant's standard operating procedure. Solutions of C10 mitoquinone mesylate (100 μg / ml) were prepared in the five solvents by diluting a solution of C10 mitochonone mesylate 1 mg / ml in water. The solutions (5 ml) were placed in glass jars, flooded with air or nitrogen, sealed and stored. Aliquots (0.25 ml) were taken at 0, 1, 2, 4, 8, 16, 32, 64 and 125 days, and the concentration of C10 mitoquinone was determined by means of HPLC.

The results are given in table 5. The stability of the mitochonone mesylate C10 in 0.01 M NaOH was not included because the mitochonone mesylate C10 decomposes in this solvent in 15 minutes. The results show that (a) the stability in solution is independent of the atmosphere over the solution; and (b) the temperature has a significant effect on the stability of the C10 mitochonone in all solvents, except HCl.

Table 5: Stability of the C10 mitoquinone mesylate in solution in 4 different solvents under different conditions Table 5 (Continued) - | o The data are averages of two values expressed as a percentage of the zero time value.

The stability of the solution of the mitochonon mesylate C10 in four solvents is also shown in figures 16, 17, 18 and 19. Stability of the mitochondone mesylate C10 in the solid state 15 The stability of the mitochonone mesylate C10 in the solid state was studied in absence of light under three different conditions: 40 ° C and 75% RH; 25 ° C and 50% RH; and 4 ° C on blue silica gel, according to the applicant's standard operating procedure. A known weight of C10 mitochonone mesylate was placed in 0 clear glass containers and stored under different conditions; Samples were taken in duplicate on days 1, 2, 4, 8, 16, 32, 64 and 125, and the concentration of mitochondone mesylate C10 was determined by HPLC after dissolving the samples in water. The results are given in table 6 and figure 20. Mitoquinone mesylate C10 was stable (< 10% decomposition) at 4 ° C on silica gel for 125 days, and at 25 ° C / 50% RH for 60 days.

Table 6: stability of the mitochonone mesylate C10 in the solid state at 40 ° C and 75% RH; 25 ° C v 50% RH: v 4 ° C on that of blue silica.

The data are the means of two values expressed as a percentage of the zero time value.

Example 9: Stability studies of the mitochondone mesylate complex C10: β-cyclodextrin (1: 2) Stability of the mitochondone mesylate complex C10: ß-cyclodextrin in solution The solution stability of the mitochonone mesylate complex C10: β-cyclodextrin (1: 2) was determined in five solvents: water, 0.01 M HCl, 0.01 M NaOH, IPB (pH 7.4) and 50% MeOH, two temperatures: 25 ° C and 40 ° C, under two atmospheric conditions: air and nitrogen, for 64 days, according to the applicant's standard operating procedure. Solutions of the mitochondone mesylate complex C10: β-cyclodextrin (1: 2) (100 μg / ml as mitochonon mesylate C10) were prepared in the five solvents, by diluting a solution supplying the mitochondone mesylate complex C10: β -cyclodextrin (1: 2) (1 mg / ml as mitochonone mesylate C10), in water. The solutions (5 ml) were placed in glass jars, flooded with air or nitrogen, sealed and stored. Aliquots (0.25 ml) were taken at 0, 1, 2, 4, 8, 16, 32, 64 and 125 days, and the concentration was determined by HPLC. The results are given in table 7 and figures 21, 22, 23 and 24. The stability of the mitochonone mesylate complex C10: β-cyclodextrin (1: 2) in 0.01 M NaOH was not included, because the mesylate complex of C10 mitoquinone: ß-cyclodextrin (1: 2) decomposes in this solvent in 15 minutes. The results show that (a) the stability in solution is independent of the atmosphere over the solution; and (b) the temperature has a significant effect on the stability of the mitochonone mesylate C10 in the 1: 2 complex with β-cyclodextrin, in all solvents except HCl.

The data are the means of two values expressed as a percentage of the zero time value.

I Stability of the mitochondone mesylate complex C10: β-cyclodextrin (1: 2) in the solid state The stability of the mitochondone meslain complex C10: β-cyclodextrin (1: 2) in the solid state was studied in the absence of light under three conditions. different conditions: 40 ° C and 75% RH; 25 ° C and 50% RH; and 4 ° C on blue silica gel, according to the applicant's standard operating procedure. A known weight of the mitochonone mesylate complex C10: β-cyclodextrin (1: 2) was placed in clear glass containers that were stored under different conditions. Samples were taken in duplicate on days 1, 2, 4, 8, 16, 32, 64 and 125, and the concentration of mitochondone mesylate C10 was determined by HPLC after dissolving the samples in water. The results are given in table 8 and figure 25. The results show that the mitochonone mesylate C10 was stable in the mltoquinone mestolate complex C10: β-cyclodextrin (1: 2) at 4 ° C on silica gel blue, and at 25 ° C and 50% RH. At 40 ° C and 75% RH, 37% of the mitochonone mesylate C10 was degraded from the mitochonone mesylate complex C10: β-cyclodextrin (1: 2) by storage for 64 days.

Table 8: Solid state stability of the mitochondone mesylate complex C10: β-cyclodextrin (1: 2) at 40 ° C v 75% RH: 25 ° C v 50% RH; and 4 ° C on blue silica gel The data are the means of two values expressed as a percentage of the zero time value. * Average of two very different values (71.9 and 31.1%) Example 10: Stability studies of the mitochondone mesylate complex C10: β-cyclodextrin (1: 1) Stability in solution The solution stability of the mitochonone mesylate complex C10: β-cyclodextrin (1: 1) was determined in five solvents: water, 0.01 M HCl, 0.01 M NaOH, IPB (pH 7.4) and 50% MeOH, at two temperatures: 25 ° C and 40 ° C, under two atmospheric conditions: air and nitrogen, for 64 days, according to the standard operating procedure of the applicant. Solutions of the mitochonone mesylate complex C10: β-cyclodextrin (1: 1) (100 μg / ml as mesylate from mitochonone C10) in the five solvents, by diluting a solution of supply of the complex mitochondrion mesylate C10: ß-cyclodextrin (1: 1) (1 mg / ml as mitochonone mesylate C10), in water. The solutions (5 ml) were placed in glass jars, flooded with air or nitrogen, sealed and stored. Aliquots (0.25 ml) were taken on days 0, 1, 2, 4, 8, 16, 32, 64 and 125, and the concentration was determined by HPLC. The results are given in Table 9 and Figures 26, 27, 28 and 29. The stability of the mitochonone mesylate complex C10: β-cyclodextrin (1: 1) in 0.01 M NaOH was not included, because the mitoquinone mesylate C10 It decomposes in this solvent in 15 minutes. The results show that (a) the stability in solution is independent of the atmosphere over the solution; and (b) the temperature has a significant effect on the stability of the C10 mitoquinone mesylate in the 1: 1 complex with β-cyclodextrin in water and IPB, but not in nl HCl in 50% MeOH.

Table 9: Solution stability of the mitochondone mesylate complex C10: β-cyclodextrin (1: 1) in 4 different solvents under different conditions Table 9 (Continued) The data are the means of two values expressed as a percentage of the zero time value.

Solid state stability The stability of the mitochondone mesylate complex C10: β-cyclodextrin (1: 1) in the solid state was studied in the absence of light under three different conditions: 40 ° C and 75% RH; 25 ° C and 50% RH; and 4 ° C on blue silica gel, according to the applicant's standard operating procedure.

A known weight of the mitochonone mesylate complex C10: β-cyclodextrin (1: 1) was placed in clear glass containers that were stored under different conditions. Samples were taken in duplicate on days 1, 2, 4, 8, 16, 32, 64 and 125, and the concentration of mitochondone mesylate C10 was determined by HPLC after dissolving the samples in water. The results are given in Table 10 and Figure 30. The results show that the mitochonone mesylate C10 was stable at 4 ° C on silica gel and at 25 ° C and 50% RH; however, by storage for 125 days at 40 ° C and 75% RH, 37% of the mitochonone mesylate C10 was degraded in the mitochonone mesylate complex C10: β-cyclodextrin (1: 1).

Table 10: Solid state stability of the mitochondone mesylate complex C10: β-cyclodextrin (1: 1) at 40 ° C and 75% RH; 25 ° C and 50% RH; and 4 ° C on blue silica gel The data are the means of two values expressed as a percentage of the zero-time value.

Example 11: Pharmacokinetic study of a single dose i.v. and oral of the mitochondone mesylate complex C10: β-cyclodextrin (1: 2) in rat (P2 and P3) Based on the results of a previous pharmacokinetic study of mitochonon bromide and an acute oral toxicity study of the mitochonone meslate complex C10: β-cyclodextrin (1: 2), the doses of the mitochondone mesylate complex C10: β -cyclodextrin (1: 2) for the pharmacokinetic study, were 50 mg / kg of C10 mitoquinone mesylate for the oral dose, and 10 mg / kg of mitochonone mesylate C10 for the iv dose Forty-eight hours before the experiment, 10 female Wistar rats (average weight of approximately 236 g) were canalized with Silastic tubing in the right jugular vein, under halothane anesthesia. An aqueous solution of the complex of mitochondone mesylate C10: β-cyclodextrin (1: 2) (10 mg / ml as mitochonon mesylate C10) was prepared and administered either orally (n = 5) or via iv (n = 5). Blood samples (0.2 ml) were taken at 0, 5, 10, 20, 30, 45, 60, 90, 120, 180, 240, 360, 720 and 1440 min (24 h) after the i.v dose; and at 0, 15, 30, 60, 90, 120, 150, 180, 240, 300, 420, 540, 720 and 1440 min (24 h) after the oral dose. The blood samples were centrifuged and the plasma samples (0.1 ml) were stored in the freezer at -20 ° C. Samples of urine and faeces were also collected at 24 h.

The concentration of C10 mitochonone mesylate in the plasma was determined by ESR using LC / MS (Table 12). Pharmacokinetic analysis The pharmacokinetics of C10 mitoquinone was analyzed by non-linear, non-weighted least-squares regression using MINIM. The data of the i.v. they were adjusted using one, two and three compartment models. The model that gave the best fit was the minimum value according to the Akaike information criterion (A.I.C.). It was found that the drug concentration curves in the plasma against the time after administration were the best, and were adjusted appropriately by means of an open three compartment model described by the following equation: C = Ae-at + Be-pt + Ee-rt where C is the concentration of the drug in the plasma, A, B and E are mathematical coefficients, is the velocity constant for the distribution phase, ß is the velocity constant for an intermediate phase (distribution or elimination), and? is the rate constant for a slower terminal phase of elimination. The elimination half-life of the drug (ty2) in the terminal phase was calculated as t% = 0.693 / ?: The data for the oral dose (after 4 h) were adjusted with a one compartment model. The peak concentration (Cmax) and the time to reach the Cma? (tma?) were obtained directly from the concentration-time profile. The area under the curve (AUC) concentration in the plasma-time was estimated using the linear trapezoidal rule, with extrapolation from the last measured concentration to infinity, determined using the terminal elimination rate constant (. Plasma after intravenous (CL) and oral administration (CIJF) was estimated as CL = dose / AUC The volumes of distribution were calculated as Vß = dose / (AUC- ß) and V? = dose / (AUO). Absolute bioavailability (F) was calculated as: F = AUCora? x DosiSj.v./AUCi.v. x DosiSorai- The mean residence time (MRT) was calculated as AUMC / AUC.The apparent volume of distribution in steady state ( Vss) was calculated as dose.v. X AUMC / (AUC) 2. Results and discussion Figures 31A-31B show the mean plasma-time concentration profiles of mitochondrion mesylate C10, after iv administration and of the mitochondone mesylate complex C10: ß- cyclodextrin (1: 2), and in Table 11 the average pharmacokinetic parameters are given. Original data of the plasma concentration of mitochonon mesylate C10 are annexed (table 12).

Table 11: Pharmacokinetic parameters of mitochonone mesylate C10 administered as a complex of C10 mitoquinone mesylate: 6-cyclodextrin (1: 2) in rat, after single doses i.v. (10 mq / kq) v oral (50 mq / kq) Mytoquinone-C10 i.v. Mitochonone-oral C10 (n = 5) (n = 5) Body weight (g) 236.8 ± 21.0 236.8 ± 22.9 Cmax (ng / ml) - 35.1 tmax (min) - 30 t? / 2? (h) 1.83 ± 0.44 - t? / 2 (h) 14.3 * 13.9 ** AUC (μg min / ml) 47.3 ± 11.1 29.3 ± 2.7 AUMC (μg min / ml) 5292 ± 831 7477 ± 365 F (%) 100 12.4 CL (l / min / kg) 0.22 ± 0.04 - CIJF (l / min / kg) - 13.7 ± 1.3 Vß (l / kg) 3.33 ± 1.46 - V? (l / kg) 24.04 ± 18.3 - MRT (h) 4.2 ± 0.5 9.5 ± 2.2 Vss (l / kg) 25.2 ± 6.5 - value of faith obtained from mean concentrations at times > 4h * value of faith obtained from mean concentrations at times > 4h After i.v. administration, a very rapid distribution phase is followed by a slower initial distribution or elimination phase, followed by approximately 4 hours of a prolonged elimination phase. The concentration-time profile of the C10 mitoquinone was adjusted to a three-compartment model with a terminal half-life of 1.8 h, although the half-life based on the data considered after 4 h of the dose is 14.3 h (Table 13 ). After oral administration, the absorption of C10 mitoquinone from the Gl tract of the rat was very rapid. The peak concentration of the C10 mitochonon in plasma occurred within 1 h after oral administration and then declined slowly over time, with an elimination half-life based on data after 4 h, approximately 14 h. The estimated F value is 12.4%. t o Lp Lp Table 13: Pharmacokinetics of the mitochondone mesylate complex C10: β-cyclodextrin (1: 2) i.v. (P ") and oral (P3) 3 compartment model Code A K1 B K2 K3 r2 A.I.C. of rat SD value SD value SD value SD value SD value SD value 0.9999 69.21 final end final end final end P2 01 i.v. 22766.1 12412.7 0.7080 0.1143 378.5 23.2 0.0456 0.0047 49.6 13.2 0.0057 0.0016 P2 02 i.v. 2868.7 1502.0 0.3945 0.1855 591.0 325.4 0.0902 0.0317 105.0 26.2 0.0100 0.0024 0.9996 85.45 P2 03 i.v. 6736.1 1535.2 0.4168 0.07638 1029.7 248.0 0.0945 0.0128 72.2 11.4 0.0067 0.0013 0.9999 77.63 P2 04 i.v. 13002.7 880.7 0.3973 0.0198 591.4 96.3 0.0639 0.0092 88.5 13.7 0.0050 0.0010 1.0000 82.32 P2 05 i.v. 9353.2 943.4 0.4073 0.0257 510.0 80.0 0.0638 0.0094 50.4 14.0 0.0060 0.0020 0.9999 77.42 Medium 10945.3 3454.8 0.4648 0.0838 620.1 154.6 0.0716 0.0136 73.1 15.7 0.0067 0.0017 0.9999 78.4 SD 7573.8 5016.9 0.1362 0.0686 244.9 126.6 0.0204 0.0105 24.1 6.0 0.0020 0.0005 0.0001 6.1 N3 • ^ 1 2 compartment model Code A K1 B K2 A.I.C. of rat SD value SD value SD value SD value 0.9990 96.46 final end final P2 01 i.v. 8722.0 2340.9 0.4962 0.0583 320.2 28.9 0.0267 0.0027 P2 02 i.v. 2131.2 135.1 0.2295 0.0158 219.5 27.7 0.0199 0.0029 0.9986 97.56 P2 03 i.v. 4787.7 303.8 0.2642 0.0173 313.8 61.8 0.0289 0.0056 0.9990 108.04 P2 04 i.v. 10519.2 755.6 0.3306 0.0165 316.4 46.5 0.0203 0.0038 0.9992 117.86 P2 05 i.v. 7711.2 581.3 0.3466 0.0187 324.2 46.0 0.0317 0.0044 0.9995 100.53 Average 6774.2 823.3 0.3334 0.0253 298.8 42.2 0.0255 0.0039 0.9991 104.1 SD 3324.2 881.7 0.1028 0.0185 44.53 14.17 0.0052 0.0012 0.0003 8.9 All patents, publications, scientific articles and other documents and materials referred to or mentioned in the present are indicative of the knowledge that is held in the matter to which the invention pertains, and each document and material referred to is incorporated in this description as reference as if it were indicated individually that it is incorporated in its entirety as a reference. Applicants reserve the right to physically incorporate into this specification each and every one of the materials and information of any such patents, publications, scientific articles, websites, electronically available information and other materials or documents cited. The specific methods and compositions described herein are representative of various preferred embodiments or modalities, and are exemplary only and are not considered to be limitations on the scope of the invention. Other objects, aspects, examples and embodiments will be considered by those skilled in the art upon reading this specification, and are encompassed within the spirit of the invention defined by the scope of the claims. It will be very apparent to those skilled in the art that various substitutions and modifications to the described invention can be made without departing from the scope and spirit of the invention. The illustratively described invention can be practiced adequately in the absence of any element or elements, or limitation or limitations, which are not specifically stated as essential. Thus, for example, in each case of the present, in embodiments or examples of the present invention, any of the terms "comprising", "consisting essentially of" or "consisting of" may be replaced with any of the other two terms in the specification. Also, the terms "comprising", "including", "containing", etc., must be considered broadly and without limitation. The methods and methods described herein illustratively can be practiced appropriately in a different order of steps, since they are not necessarily restricted to the order of steps indicated herein or in the claims. Also, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references, unless the context clearly dictates otherwise. A) Yes, for example, a reference to "a host cell" includes a plurality of such host cells (e.g., a culture or population), and so forth. Under no circumstances may the patent be construed as limited to the specific examples or embodiments or methods specifically described herein. Under no circumstances may the patent be construed as limited by any statement made by any Examiner or other official or employee of the Patent and Trademark Office, unless such affirmation is expressly adopted, specifically and without qualification or reservation, in a written respondent of the applicants. The terms and expressions used are used as terms of description and not limitation, and the use of such terms and expressions is not intended to exclude any equivalent of the characteristics shown and described or portions thereof.; however, it is recognized that several modifications are possible within the scope of the claimed invention. Thus, it will be understood that while the present invention has been specifically described by preferred embodiments and optional features, those skilled in the art can modify and vary the concepts described herein, and that such modifications and variations are considered to be within the scope of this invention. defined by the appended claims. The invention has been described broadly and generically in the present. Each of the subgeneric species and sets that are within the generic description also forms part of the invention. This includes the generic description of the invention, with a negative condition or limitation to remove any matter of the genre, without considering whether the separate material is specifically cited here. Other embodiments are within the following claims. In addition, where the features or aspects of the invention are described in terms of the Markush groups, those skilled in the art will recognize that the invention is also described as such in terms of any individual member or subgroup of members of the Markush group. The compounds of the invention have application in selective antioxidant therapies in human patients to prevent mitochondrial damage. This may be to prevent elevated mitochondrial oxidative stress associated with particular diseases, such as Parkinson's disease, or diseases associated with mitochondrial DNA mutations. They could also be used in conjunction with cell transplant therapies for neurodegenerative diseases, to increase the survival of implanted cells. In addition, these compounds could be used as prophylactics to protect the organs during a transplant, or correct the ischemia-reperfusion injury that occurs during surgery. The compounds of the invention could also be used to reduce cell damage after a stroke or heart attack, or they can be given prophylactically to premature babies who are susceptible to cerebral ischemia. The methods of the invention have a major advantage over current antioxidant therapies - they will allow the antioxidants to accumulate selectively in the mitochondria, the part of the cell under the greatest oxidative stress. This will greatly increase the effectiveness of antioxidant therapies. Those skilled in the art will appreciate that the above description is given by way of example only and that different combinations of lipophilic cation / antioxidant can be employed without departing from the scope of the invention.

Claims (115)

1. NOVELTY OF THE INVENTION CLAIMS 1. - A compound comprising a lipophilic cationic portion bound by a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mltochondria, and the of salt is chemically stable or the anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion or the binding portion.
2. 2. A stable compound comprising a lipophilic cationic portion bound by means of a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and wherein the anionic complement is not a halogen ion, and the anionic complement is non-nucleophilic, or the anionic complement exhibits no reactivity against the cationic portion, the binding portion, or the antioxidant portion.
3. 3. The compound according to claim 1 or 2, further characterized in that the antioxidant portion is a quinone or a quinol.
4. 4. The compound according to claim 1 or 2, further characterized in that the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives, chain-breaking antioxidants, including butylated hydroxyanisole, butylated hydroxytoluene, sweeteners radicals in general, including modified fullerenes, spin traps including derivatives of 5,5-dimethylpyrroline N-oxide, ér-butylnitrosobenzene, ér-nitrosobenzene, α-phenyl-fer-butylnitrone, and related compounds.
5. 5. The compound according to any of claims 1 to 4, further characterized in that the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation.
6. 6. The compound according to claim 5, further characterized in that it has the general formula I: or its quinol form, wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of Ci to C5 (optionally substituted) or H, and wherein n is an integer from about 2 to about 20 , and where Z is a non-reactive anion.
7. 7. The compound according to claim 6, further characterized in that Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates.
8. 8. The compound according to claim 6 or claim 7, further characterized in that the C of the bridge (C) n is saturated.
9. 9. The compound according to claim 8, further characterized in that it has the formula: II or its quinol form, wherein Z is a non-nucleophilic anion.
10. 10. The compound according to claim 9, further characterized by having the formula: (lil) 11. A pharmaceutical composition comprising or including a compound comprising a lipophilic cationic moiety bound by a binding portion to an antioxidant portion, and an anionic complement for said cationic moiety, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement does not exhibit reactivity against the antioxidant portion, the cationic portion, or the binding portion. 12. A pharmaceutical composition comprising or including a stable compound comprising a lipophilic cationic moiety bound by a binding portion to an ampho-oxidant portion, and an anionic complement to said cationic moiety, wherein the cationic species is capable of directing antioxidant species to the mitochondria, and wherein the anionic complement is not a halogen ion, and the anionic complement is non-nucleophilic or the anionic complement does not exhibit reactivity against the cationic portion, the binding portion, or the antioxidant portion. 13. - The composition according to claim 11 or 12, further characterized in that the antioxidant portion is a quinone or a quinol. 14. The composition according to claim 11 or 12, further characterized in that the ampho-oxidant portion is selected from the group comprising vitamin E and vitamin E derivatives, chain-breaking ampho-oxidants, including butylated hydroxlanisol, butylated hydroxytoluene, sweeteners radicals in general, including modified fullerenes, spin traps including derivatives of 5,5-dimethylpyrroline N-oxide, ér-butylnitrosobenzene, fer-nitrosobenzene, α-phenyl-tert-butylnitrone, and related compounds. 15. The composition according to any of claims 11 to 14, further characterized in that the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation. 16. The composition according to claim 15, further characterized in that the compound has the general formula I: or its quinol form, wherein R-, R2 and R3, which may be the same or different, are selected from alkyl portions of Ci to C5 (optionally substituted) or H, and wherein n is an integer of about 2 to 20, and where Z is a non-reactive anion. 17. The composition according to claim 16, further characterized in that Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. 18. The composition according to claim 16 or claim 17, further characterized in that the C of the bridge (C) n is saturated. 19. The composition according to claim 18, further characterized in that the compound has the formula: II or its quinol form, where Z is a non-nucleophilic anion. 20. The composition according to any of claims 11 to 19, further characterized in that it comprises cyclodextrin. 21. - The composition according to claim 20, further characterized in that the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10. 22. The composition according to claim 21, further characterized in that the molar ratio of compound to cyclodextrin is from about 5: 1 to about 1: 5. 23. The composition according to claim 22, further characterized in that the molar ratio of compound to cyclodextrin is from about 4: 1 to about 1: 4. 24. The composition according to claim 23, further characterized in that the molar ratio of compound to cyclodextrin is from about 2: 1 to about 1: 2. 25. The composition according to claim 24, further characterized in that the molar ratio of compound to cyclodextrin is about 1: 1. 26. The composition according to claim 24, further characterized in that the molar ratio of compound to cyclodextrin is about 1: 2. 27.- The composition according to any of claims 20 to 26, further characterized in that the compound has the formula: (lll) 28.- The composition according to any of claims 11 to 27, further characterized in that the cyclodextrin is β-cyclodextrin. 29. The composition according to claim 28, further characterized in that the compound has the formula: (III) and the molar ratio of compound to cyclodextrin is about 1: 2. 30. The composition according to any of claims 11 to 29, further characterized in that it is formulated for oral administration. 31. The composition according to any of claims 11 to 29, further characterized because it is formulated for parenteral administration. 32.- A unit dose comprising or including a compound comprising a lipophilic cationic portion bound by a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion, or the binding portion, together with any pharmaceutically acceptable diluent, carrier or excipient. 33.- A unit dose comprising or including a stable compound comprising a bound lipophilic cationic portion by means of a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing antioxidant species to the mitochondria, and wherein the anionic complement is not a halogen ion, and the anionic complement is non-nucleophilic or the anionic complement does not exhibit reactivity against the cationic portion, the binding portion, or the antioxidant portion. 34. - The unit dose according to claim 32 or 33, further characterized in that the antioxidant portion is a quinone or a quinol. 35.- The unit dose according to claim 32 or 33, further characterized in that the antioxidant portion is selected from the group comprising vitamin E and vitamin E derivatives, chain-breaking antioxidants, including butylated hydroxyanisole, butylated hydroxytoluene, sweepers of radicals in general, including modified fullerenes, spin traps including derivatives of 5,5-dimethylpyrroline N-oxide, ér-butylnitrosobenzene, ter-nitrosobenzene, α-phenyl-tert-butylnitrone, and related compounds. 36.- The unit dose according to any of claims 32 to 35, further characterized in that the lipophilic cationic portion is a substituted or unsubstituted triphenylphosphonium cation. 37.- The unit dose according to claim 36, further characterized in that the compound has the general formula I: or its quinol form, wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of Ci to C5 (optionally substituted) or H, and wherein n is an integer of about 2 to 20 , and where Z is a non-reactive anion. 38.- The unit dose according to claim 37, further characterized in that Z is selected from the group consisting of alkyl or aryl sulfonates or nitrates. 39.- The unit dose according to claim 37 or claim 38, further characterized in that the C of the bridge (C) n is saturated. 40.- The unit dose according to claim 39, further characterized in that the compound has the formula: II or its quinol form, wherein Z is a non-nucleophilic anion. 41.- The unit dose according to any of claims 32 to 40, further characterized in that it comprises cyclodextrin. 42. - The unit dose according to claim 41, further characterized in that the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10. 43.- The unit dose according to claim 42, further characterized in that the molar ratio of compound to cyclodextrin is from about 5: 1 to about 1: 5. 44. The unit dose according to claim 43, further characterized in that the molar ratio of compound to cyclodextrin is from about 4: 1 to about 1: 4. 45.- The unit dose according to claim 44, further characterized in that the molar ratio of compound to cyclodextrin is from about 2: 1 to about 1: 2. 46.- The unit dose according to claim 45, further characterized in that the molar ratio of compound to cyclodextrin is about 1: 1. 47.- The unit dose according to claim 45, further characterized in that the molar ratio of compound to cyclodextrin is approximately 1: 2. 48. The unit dose according to any of claims 40 to 47, further characterized in that the compound has the formula: (III) 49. The unit dose according to any of claims 41 to 48, further characterized in that the Cyclodextrin is β-cyclodextrin. 50.- The unit dose according to claim 49, further characterized in that the compound has the formula: (III) and the molar ratio of compound to cyclodextrin is about 1: 2. 51.- The unit dose according to any of claims 32 to 50, further characterized because it is formulated for oral administration. 52. The unit dose according to any of claims 32 to 50, further characterized because it is formulated for parenteral administration. 53. The compound according to any of claims 1 to 10, or a pharmaceutically acceptable salt thereof, for use in the prophylaxis or treatment of oxidative stress in a mammal by administering the compound or its salt to said mammal. The compound according to any of claims 1 to 10, or a pharmaceutically acceptable salt thereof, for use in the prophylaxis or treatment of the symptoms of aging in a mammal, by administration of the compound or its salt to said mammal. The compound or pharmaceutically acceptable salt thereof according to claim 53 or 54, further characterized in that said administration is, on the first day, at a dose of about 1.02 to about 2.0 times the daily maintenance dose, followed by administration of the compound or its salt at the daily maintenance dose on subsequent days. 56. The compound or pharmaceutically acceptable salt thereof according to any of claims 53 to 55, further characterized by having the formula: or its form of quinol, where Z is a non-nucleophilic anion. The compound or pharmaceutically acceptable salt thereof according to any of claims 53 to 56, further characterized in that the salt is the methanesulfonate salt. 58.- The compound or pharmaceutically acceptable salt thereof according to any of claims 53 to 56, further characterized by combining with cyclodextrin. 59. The compound according to claim 58, further characterized in that it has the formula: (III) wherein the cyclodextrin is β-cyclodextrin and the molar ratio of compound to cyclodextrin is about 1: 2. 60.- A suitable unit dose for oral administration, comprising as active ingredient a compound as claimed in any of claims 1 to 10, the compound being or being formulated as a crystalline form or a non-liquid form. 61.- A suitable unit dose for parenteral administration comprising as active ingredient a compound as claimed in any of claims 1 to 10. 62.- A pharmaceutical composition suitable for the treatment of a patient who would benefit from a reduction of oxidative stress or of a reduction of aging symptoms, comprising or including an effective amount of a compound as claimed in any of claims 1 to 10, in combination with one or more pharmaceutically acceptable vehicles, excipients or diluents acceptable 63.- The composition according to claim 62, further characterized in that the compound is a compound of formula I: or its quinol form, wherein Ri, R2 and R3, which may be equal or different, are selected from alkyl portions of Ci to Cs (optionally substituted) or H, and wherein n is an integer of about 2 to 20, and where Z is a non-reactive anion. 64.- The composition according to claim 63, further characterized in that the compound has the formula: or its quinol form, where Z is a non-nucleophilic anion. 65. - The composition according to any of claims 62 to 64, further characterized in that it comprises cyclodextrin. 66.- The composition according to claim 65, further characterized in that the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10. 67.- The composition according to claim 66, further characterized in that the molar ratio of compound to cyclodextrin is from about 5: 1 to about 1: 5. The composition according to claim 67, further characterized in that the molar ratio of compound to cyclodextrin is from about 4: 1 to about 1: 4. 69.- The composition according to claim 68, further characterized in that the molar ratio of compound to cyclodextrin is from about 2: 1 to about 1: 2. The composition according to claim 69, further characterized in that the molar ratio of compound to cyclodextrin is about 1: 1. 71. The composition according to claim 69, further characterized in that the molar ratio of compound to cyclodextrin is about 1: 2. 72. The composition according to any of claims 65 to 71, further characterized in that the compound has the formula: (lll) 73. The composition according to any of claims 65 to 72, further characterized in that the cyclodextrin is β-cyclodextrin. 74.- The composition according to claim 73, further characterized in that the compound has the formula: (III) and the molar ratio of compound to cyclodextrin is about 1: 2. 75. The composition according to any of claims 61 to 74, further characterized in that it is formulated for oral administration. 76. The composition according to any of claims 61 to 74, further characterized because it is formulated for parenteral administration. 77.- A method for reducing oxidative stress in a cell, comprising the step of contacting said cell with a compound comprising a bound lipophilic cationic portion by means of a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion or the binding portion. 78.- A method for reducing oxidative stress in a cell, comprising the step of contacting said cell with a stable compound comprising a bound lipophilic cationic portion by means of a binding portion to an antioxidant portion, and a complement anionic for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and wherein the anionic complement is not a halogen, and the anionic complement is non-nucleophilic, or the anionic complement does not exhibit reactivity against the cationic portion, the binding portion, or the antioxidant portion. 79. - The method according to claim 77 or 78, further characterized in that the compound is a compound of formula I: or its quinol form, wherein Ri, R and R3, which may be the same or different, are selected from alkyl portions of Ci to C5 (optionally substituted) or H, and wherein n is an integer of about 2 to 20, and where Z is a non-reactive anion. 80.- The method according to claim 79, further characterized in that the compound has the formula: or its quinol form, where Z is a non-nucleophilic anion. 81. - The method according to any of claims 77 to 80, further characterized in that the compound is complexed with cyclodextrin. 82. The method according to claim 81, further characterized in that the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10. 83. The method according to claim 82, further characterized in that the molar ratio of compound to cyclodextrin is from about 5: 1 to about 1: 5. 84. The method according to claim 83, further characterized in that the molar ratio of compound to cyclodextrin is from about 4: 1 to about 1: 4. 85. The method according to claim 84, further characterized in that the molar ratio of compound to cyclodextrin is from about 2: 1 to about 1: 2. 86. The method according to claim 85, further characterized in that the molar ratio of compound to cyclodextrin is about 1: 1. 87. The method according to claim 85, further characterized in that the molar ratio of compound to cyclodextrin is about 1: 2. 88. The method according to any of claims 81 to 87, further characterized in that the compound has the formula: (lll) 89. The method according to any of claims 81 to 88, further characterized in that the cyclodextrin is β-cyclodextrin. 90. The method according to claim 89, further characterized in that the compound has the formula: (III) and the molar ratio of compound to cyclodextrin is about 1: 2. 91. - The use of a compound comprising a bound lipophilic cationic portion by means of a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion or the binding portion, in the preparation or manufacture of a drug, unit dose or pharmaceutical composition, effective for stress reduction oxidative in a patient. 92.- The use of a compound comprising a lipophilic cationic portion bound by means of a binding portion to an amphotericizing portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion or the binding portion, in the preparation or manufacture, together with one or more other materials, of a medicament, unit dose or pharmaceutical composition, effective for reducing the symptoms of aging in a patient. 93. The use claimed in claim 91, wherein the composition is a composition as claimed in any of claims 11 to 31. 94. - The use claimed in claim 91, wherein the dosage form is a dosage form as claimed in any of claims 32 to 52. 95.- The use claimed in claim 91 or 92 , wherein the antioxidant compound directed to the mitochondria is a compound of formula I: or its quinol form, wherein Ri, R2 and R3, which may be the same or different, are selected from alkyl portions of Ci to C5 (optionally substituted) or H, and wherein n is an integer of about 2 to 20 , and where Z is a non-reactive anion. 96. The use claimed in claim 95, wherein the compound is complexed with cyclodextrin. 97. The use claimed in claim 96, wherein the molar ratio of compound to cyclodextrin is from about 10: 1 to about 1: 10. 98. - The use claimed in claim 97, wherein the molar ratio of compound to cyclodextrin is from about 5: 1 to about 1: 5. 99. The use claimed in claim 98, wherein the molar ratio of compound to cyclodextrin is from about 4: 1 to about 1: 4. 100.- The use claimed in claim 99, wherein the molar ratio of compound to cyclodextrin is from about 2: 1 to about 1: 2. 101. The use claimed in claim 100, wherein the molar ratio of compound to cyclodextrin is about 1: 1. 102. The use claimed in claim 100, wherein the molar ratio of compound to cyclodextrin is about 1: 2. 103. The use claimed in any of claims 95 to 102, wherein the compound has the formula: (lll) 104. - The use claimed in any of claims 95 to 103, wherein the cyclodextrin is β-cyclodextrin. 105.- The use claimed in claim 104, wherein the compound has the formula: (III) and the molar ratio of compound to cyclodextrin is about 1: 2. 106. The use claimed in any of claims 91 to 105, wherein said composition or unit dose is formulated for oral administration. 107. The use claimed in any of claims 91 to 105, wherein said composition or unit dose is formulated for parenteral administration. 108.- The use of a compound comprising a lipophilic cationic portion bound by means of a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement exhibits no reactivity against the amphotericin portion, the cationic portion or the binding portion, in the preparation or manufacture of an effective composition for the reduction of oxidative stress in a cell. 109.- A method of synthesis of a compound of the formula I: (or its quinol form), wherein Ri, R2 and R3, which may be equal or different, are selected from alkyl portions of Ci to C5 (optionally substituted), and wherein n is an integer of about 2 to 20, said method including or comprising mixing with cyclodextrin. 110.- A method of synthesis of a compound having the formula: (III) said method including or comprising mixing with cyclodextrin. 111.- A method of synthesis of a compound that has the formula: essentially as described herein. 112.- A pharmaceutical composition suitable for the treatment of a patient suffering from or predisposed to suffering from Parkinson's disease, Alzheimer's disease, Hunfington's chorea, or Friedreich's ataxia, which comprises or includes an effective amount of a compound that comprises a lipophilic cationic moiety bound by a binding portion to an antioxidant portion, and an anionic complement to said cationic moiety, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically The stable or anionic complement exhibits no reactivity against the antioxidant portion, the cationic portion or the binding portion, in combination with one or more pharmaceutically acceptable carriers, excipients or diluents. 113.- The composition according to claim 112, characterized also because it is suitable for the treatment of a patient suffering from or predisposed to suffer from Friedreich's ataxia. 114. The use of a compound comprising a bound lipophilic cationic portion by means of a binding portion to an antioxidant portion, and an anionic complement for said cationic portion, wherein the cationic species is capable of directing the antioxidant species to the mitochondria, and the salt form is chemically stable or the anionic complement does not exhibit reactivity against the antioxidant portion, the cationic portion or the binding portion, in the preparation or manufacture of a medicament, unit dose, or pharmaceutical composition, effective for the treatment or prophylaxis of a patient suffering from or predisposed to suffering from Parkinson's disease, Alzheimer's disease, Hunfington's chorea, or Friedreich's ataxia. 115. The use claimed in claim 114, wherein the medicament, unit dose, or pharmaceutical composition is effective for the treatment or prophylaxis of a patient suffering from or predisposed to undergoing Friedreich ataxia.