Mitochondria-targeted ubiquinone derivatives as oxidative stress reducing antioxidants, pharmaceutical composition, method of production and method of treatment

FIELD: chemistry.

SUBSTANCE: invention relates to a novel chemically stable antioxidant compound which contains a lipophilic cationic moiety linked by a linking moiety to an antioxidant molecule and an anionic component for said cationic moiety, where the antioxidant compound is a mitoquinone, selected from: 10-(6'-ubiquinone)propyltriphenylphosphonium, 10-(6'-ubiquinonyl)pentyltriphenylphosphonium, 10-(6'-ubiquinonyl)decyltriphenylphosphonium, and 10-(6'-ubiquinonyl)pentadecyltriphenylphosphonium, having general formula I: or quinol form thereof, where R1, R2 and R3 denote CH3, the C atom in (C)n is saturated and n equals 3, 5, 10 or 15, and Z denotes an anionic component which is selected from a group consisting of methanesulphonate and ethanesulphonate. The invention also relates to a pharmaceutical composition for reducing oxidative stress in a cell, containing said compound and optionally containing β-cyclodextrin, a method of reducing oxidative stress in a cell and a method of producing an antioxidant compound.

EFFECT: improved properties of compounds.

25 cl, 31 dwg, 13 tbl, 11 ex

 

The technical field to which the invention relates.

The present invention relates to amphiphilic antioxidant compounds containing lipophilic cationic group, synthesis, composition and physico-chemical properties of these compounds, which favor their use, for example, as pharmaceuticals.

Prior art

Oxidative stress contributes to a number of degenerative diseases associated with aging, such as Parkinson's and Alzheimer's, and Huntington's chorea and hereditary ataxia, and non-specific disorders that accumulate with age. It also promotes inflammation and ischemia-reperfusion tissue damage in stroke and heart attack, as well as in the process of organ transplantation and surgical treatment. To prevent disturbance caused by oxidative stress, has developed a number of antioxidant therapies. However, most of them are not targeted to the cells, and therefore are less than optimally effective. Moreover, many of these antioxidants have adverse physico-chemical properties that limit, for example, their bioavailability and their ability to penetrate into the target organ and perform therapeutic action is selected.

Mitochondria are the intracellular organelles responsible for energy metabolism. Accordingly, mitochondrial defects are damaging, especially to the nervous and muscular tissues with high energy demand. They are also the main source of free radicals and reactive oxygen-containing fragments, which cause oxidative stress within most cells. Therefore, applicants believe that the delivery of antioxidants selectively into mitochondria will be more effective than the use of non-input targeted antioxidants. Accordingly, the present invention relates to the availability of antioxidants that can be delivered to the mitochondria.

Lipophilic cations can accumulate in the mitochondrial matrix due to its positive charge (Rottenberg, 1979, Methods Enzymol., 55, 547. Chen, 1988, Ann. Rev. Cell Biol., 4,155). Such ions can be accumulated, provided that they are sufficiently lipophilic to screen the positive charge or delocalizing it on a large surface area, and provided that there is no active leaks, and cation participates in metabolism or immediately does not poison the cell.

Therefore, the present invention is directed to the development approach, with p the power which you can use the ability of mitochondria to concentrate specific lipophilic cations and fix associated antioxidants, so that the target image to send an antioxidant to the main source of free radicals and reactive oxygen-containing fragments, causing oxidative stress.

Examples of antioxidants that have shown good antioxidant capacity in vivo, and showed weak antioxidant activity in relation to the separation of the target in vivo, include Coenzyme Q (CoQ) and Idebenone. Both of these compounds have low bioavailability and should be introduced at a very high dose to be effective, and therefore have a low therapeutic efficacy in comparison with the dose.

Not wishing to be bound to any theory, the authors believe that the connection antioxidant activity in vivo or ex vivo (e.g., antioxidant activity or mitochondrial accumulation) in any way is not the only factor determining the functionality of an antioxidant and/or efficacy in vivo (such as the effectiveness of therapeutic action). Although it is indeed true that in order to be used as directed insertion in mitochondria antioxidant compounds of the present invention, the antioxidant compound to be suitable antioxidant effect in vivo or ex vivo to be effective in vivo, targeted insertion in mitochondria ant the oxidant compound must exhibit other desirable physical and chemical properties, such as, for example, a suitable bioavailability suitable localization or distribution within the mitochondria-target and/or a suitable stability.

Not wishing to be bound to any theory, the inventors believe that the directed insertion in mitochondria antioxidant compounds of the present invention are the predominant antioxidant effects, including bioavailability and/or directed introduction to mitochondria and accumulation in vivo, at least partially due to their physico-chemical properties, such as, for example, their amphiphiles, their physical structure and/or size, their hydrophobicity from low to medium and/or the distribution coefficient. Therefore, such compounds are therapeutically effective at low dosages in comparison with other antioxidant compounds.

In U.S. patent No. 6331532 in the explanatory examples of compounds methanol and mitogenome (named United as methanol/mitchison) disclosed perspective directional introduction mitochondria antioxidant molecules associated with a lipophilic cation covalently coupled with one molecule of antioxidant. This patent as an example of the connection (despite the General view of the length colophony ties) is a compound of mitogenome formula

length uglev the burly chain of 10 carbon atoms (i.e. bridge10). Its reduced form, methanol, also contains10bridge.

The inventors have found that metochion/methanol, despite excellent antiokislitelnoe action, direction, and nakoplennye in mitochondria in vitro and in vivo appeared to be somewhat unstable in the form of bromide. The inventors have also found that the physico-chemical properties of mitogenome/methanol, as disclosed in U.S. patent No. 6331532, less suitable for the pharmaceutical composition, for example, when the introduction is carried out orally or parenterally, and/or when directed introducing the compound into the mitochondria in the tissue of internal organs (e.g. brain, heart, liver, or other organs).

Examples of compounds of the present invention are suitable for use in pharmaceutical compositions. They must be in a form different from the crystal, and/or solid form, but are subject to transformation in solid form when mixed with other agents, such as, for example, carriers, fillers, complexing agents or other additives and the like, for example, cyclodextrins. Mostly these agents are pharmaceutically acceptable.

The inventors have appreciated the need to offer examples of amphiphilic, directed insertion in mitochondria antioxidant is connected to the second present invention with their positive charge in combination with a suitable anion, to ensure the connection is in the form of a General neutralized salts, including solid or crystalline products. However, the inventors have found that in the forms of such salts of some salt-forming anions are best avoided, as they are reactive to the anti-oxidant compound, for example, a molecule of antioxidant binding fragment or lipophilic cationic fragment, and/or may lead to the splitting of a molecule of antioxidant or removal from it. Other soleobrazutaya anions are considered pharmaceutically undesirable. For example, nitrate residues are considered pharmaceutical companies in General are not suitable and pharmaceutically or environmentally unacceptable, whereas bromovalerate, often used for salt formation of such compounds, possesses nucleophilic properties, which can lead to the emergence of reactionary activity towards molecules of an antioxidant, for example, the cleavage of a methyl group from a molecule of antioxidant compounds of General formula (II) and/or some General decrease of the stability of the whole connection. For example, the authors determined that Hydrobromic salt mitogenome somewhat unstable.

Therefore, the inventors believe that the form of salts, including salts as liquid, solid or crystalline form, the direction of the input but in the mitochondria of antioxidants is best associated with the anion or a similar fragment, which is not nucleophilic and/or anion, which has no reaction activity with respect to any of the fragments, including antioxidant compound or complex. Also preferably, the anion was pharmaceutically acceptable.

The purpose of the invention

Accordingly, the present invention relates to amphiphilic pharmaceutically acceptable antioxidant compounds and compositions, dosage forms and methods associated with these compounds, which can be used, for example, in the treatment of diseases or conditions associated with oxidative stress, or to provide the public with valuable choice.

Summary of the invention

In the first aspect of the present invention relates to a compound containing lipophilic cationic fragment associated connecting with the fragment of the molecule antioxidant, and anionic component for the specified cationic fragment, and cationic fragment can be directed to enter into the mitochondria antioxidant molecules, and form salts are chemically stable and/or anionic component has no reaction activity to the molecule antioxidant, cationic portion or the connecting portion.

In one embodiment, the molecule antioxidant is a quinone or China.

Another is ariante implementation molecule antioxidant selected from the group including vitamin b and derivatives of vitamin b, antioxidants circuit, including mutilateral hydroxyanisol, mutilateral hydroxytrol total acceptors radicals, including derivateservlet fullerenes, spin traps, including derivatives of 5,5-dimethylpyrrole-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

In one embodiment, a lipophilic cationic fragment is substituted or unsubstituted cation triphenylphosphane.

In one embodiment, the compound has a General formula I

and/or its hinolinol form, in which groups R1, R2and R3that may be the same or different, selected From1-C5alkyl (possibly substituted) groups or N, in which n means an integer from about 2 to about 20, and in which Z signifies directionspanel anion.

Preferably Z is selected from the group comprising alkyl - or arylsulfonate or nitrates.

Preferably each link With-in bridge (S)nis saturated.

In a preferred embodiment, the compound has the formula

and/or its hinolinol form, where Z means a non-nucleophilic anion.

More prefer is Ino the compound has the formula

In another aspect the invention relates to pharmaceutical compositions containing or comprising a compound containing lipophilic cationic fragment, a linked connection with the fragment of the molecule antioxidant, and anionic component for the specified cationic fragment, and the cationic group is able to drive in mitochondria antioxidant molecules and form salt is chemically stable and/or anionic component does not show the reaction activity with respect to the molecule antioxidant, cationic portion or the connecting portion.

In one embodiment, the molecule antioxidant is a quinone or China.

In other embodiments, the implementation of a molecule of an antioxidant selected from the group comprising vitamin E and derivatives of vitamin E, antioxidants decay chain, including mutilateral hydroxyanisol, mutilateral hydroxytrol total acceptors radicals, including derivateservlet fullerenes, spin traps, including derivatives of 5,5-dimethylpyrrole-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

In one embodiment, a lipophilic cationic group is a substituted or unsubstituted cation triphenylphosphane.

One in which the version of the implementation, the compound has the General formula I.

and/or its hinolinol form, where the group R1, R2and R3that may be the same or different, selected From1-C5alkyl (possibly substituted) groups or N, where n means an integer from about 2 to about 20, and where Z means directionspanel anion.

Preferably Z is selected from the group comprising alkyl - or arylsulfonate or nitrates.

Preferably each link carbon-carbon (C)n the bridge is full.

In an additional embodiment, the compound has the formula

and/or its hinolinol form, where Z represents a non-nucleophilic anion.

In another embodiment, the composition includes a compound having the formula II, and/or hinolinol form, where Z is not nucleophilic anion, and where the composition comprises a cyclodextrin.

In various examples, the molar ratio of the 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, from about 2:1 to about 1:2 or about 1:1, for example, the molar ratio of the compound to cyclodextrin is about 1:2.

More preferably, the composition includes a compound having the formula

where qi is latextrem is a β-cyclodextrin, more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In one embodiment, the pharmaceutical composition is formulated for oral administration.

In an additional embodiment, the pharmaceutical composition is formulated for parenteral administration.

In an additional aspect, the present invention relates to a dosage form containing or comprising a compound containing lipophilic cationic fragment associated connecting with the fragment of the molecule antioxidant, and anionic component for the specified cationic fragment, and cationic fragments is able to enter into the mitochondria antioxidant molecules and form salt is chemically stable and/or anionic component does not show the reaction activity with respect to the molecule antioxidant, cationic portion or the connecting portion, together with any pharmaceutically acceptable diluent and/or carrier and/or excipient.

In one embodiment, the molecule antioxidant is a quinone or China.

In other embodiments, the implementation of a molecule of an antioxidant selected from the group comprising vitamin E and derivatives of vitamin E, antioxidants decay chain including mutilateral hydroxyanisol, mutilateral hydrox the toluene, total acceptors radicals, including derivateservlet fullerenes, spin traps, including derivatives of 5,5-dimethylpyrrole-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

In one embodiment, a lipophilic cationic fragment represents a cation substituted or unsubstituted triphenylphosphane.

In one embodiment, the compound has a General formula I

and/or its hinolinol form, where the group R1, R2and R3that may be the same or different, selected From1-C5alkyl (possibly substituted) groups or N, where n means an integer from about 2 to about 20, and where the symbol Z indicates directionspanel anion.

Preferably, the symbol Z is selected from the group comprising alkyl - or arylsulfonate or nitrates.

Preferably each link carbon-carbon (C)n the bridge is full.

In an additional embodiment, the compound has the formula and/or its hinolinol form, where Z is not nucleophilic anion.

In an additional embodiment, dosage form contains a compound having the formula II, and/or hinolinol form, where Z is not nucleophilic anion, and where the composition of the ash is separated cyclodextrin.

In various examples, the molar ratio of the 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, from about 2:1 to about 1:1, for example, the molar ratio of the compound to cyclodextrin is about 1:2.

More preferably the dosage form contains a compound having the formula

where the cyclodextrin is a β-cyclodextrin, more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In one embodiment, dosage form suitable suitable for oral administration.

In an additional embodiment, a pharmaceutical form suitable suitable for parenteral administration.

In an additional aspect, the present invention relates to the compound or its pharmaceutically acceptable salt, compositions or dosage form of the present invention for use in the prevention or treatment of oxidative stress in mammals by introducing the compound or its salts specified mammal.

In one embodiment, the compound is a compound of formula II or its pharmaceutically acceptable salt.

In another embodiment, the specified conduct introduction at first is the tier at the dosage, exceeding the daily dose of approximately from 1,02 to about 2,0 times, followed by the introduction of compounds or salts thereof in a daily dose in the next days.

Preferably the salt is a salt methanesulfonate, and the connection is combined with a cyclodextrin.

A more preferred compound has the formula

Preferably, the cyclodextrin is β-cyclodextrin, more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In an additional aspect, the present invention relates to the compound or its pharmaceutically acceptable salt, compositions or dosage form of the present invention for use in preventing or treating symptoms of aging in mammals by introducing the compound or its salts specified mammal.

In one embodiment, the compound is a compound of formula II or its pharmaceutically acceptable salt.

In another embodiment, the specified conduct introduction on the first day in a dosage that is higher than the daily dose is about 1,02-2.0 times, followed by the introduction of compounds or salts thereof in a daily dose in the next days.

Preferably the salt is a salt methanesulfonate, and the connection is combined with a cyclodextrin.

More preferably the compound has the formula

Preferably, the cyclodextrin is β-cyclodextrin, more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In an additional aspect, the present invention relates to a stable connection, including lipophilic cationic fragment associated connecting with the fragment of the molecule antioxidant, and anionic component for the specified cationic fragment, where

cationic fragment is able to enter the mitochondria-molecule antioxidant, and

anionic component is not halogen ion, and

anionic component is not nucleophilic and/or

anionic component does not show the reaction activity relative to the cationic fragment of the connecting fragment or molecule antioxidant.

In one embodiment, the molecule antioxidant is a quinone or China.

In another embodiment, the molecule of antioxidant selected from the group comprising vitamin E or derivatives of vitamin E, an antioxidant circuit, including mutilateral hydroxyanisol, mutilateral hydroxytrol total acceptors radicals, including derivateservlet fullerenes, spin traps, including derivatives of 5,5-dim telperion-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

In one embodiment, a lipophilic cationic fragment cation is substituted or unsubstituted triphenylphosphane.

In one embodiment, the compound has a General formula I

and/or its hinolinol form, where the group R1, R2and R3that may be the same or different, selected from C1-C5alkyl (possibly substituted) groups or N, where n means an integer from about 2 to about 20, and where the symbol Z indicates directionspanel anion.

Preferably Z is selected from the group comprising alkyl - or arylsulfonate or nitrates.

Preferably each link carbon-carbon (C)n the bridge is full.

In a preferred embodiment, the compound has the formula

and/or its hinolinol form, where Z is not nucleophilic anion.

More preferably the compound has the formula

In another aspect the invention relates to pharmaceutical compositions containing or comprising a stable connection involving cationic fragment, which is a lipophilic cationic fragment related connective fragm Tom with one molecule of antioxidant, and anionic component for the specified cationic fragment, and

cationic fragment is able to enter into the mitochondria antioxidant molecules and

anionic component is not a halogen ion, and

anionic component is not nucleophilic and/or

anionic component does not show the reaction activity in relation to the cationic portion, the connecting fragment or molecule antioxidant.

In one embodiment, the molecule antioxidant is a quinone or China.

In other embodiments, the implementation of a molecule of an antioxidant selected from the group comprising vitamin E or derivatives of vitamin E, antioxidants circuit, including mutilateral hydroxyanisol, mutilateral hydroxytrol total acceptors radicals, including derivateservlet fullerenes, spin traps, including derivatives of 5,5-dimethylpyrrole-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

In one embodiment, a lipophilic cationic fragment represents a cation substituted or unsubstituted triphenylphosphane.

In one embodiment, the compound has a General formula I

and /or its hinolinol form, where the group R1, R2and R3to the which may be the same or different, selected from C1-C5alkyl (possibly substituted) groups or N, where n means an integer from about 2 to about 20, and where the symbol Z indicates directionspanel anion.

Preferably Z is selected from the group comprising alkyl - or arylsulfonate or nitrates.

Preferably each link carbon-carbon (C)n the bridge is full.

In an additional embodiment, the compound has the formula

and/or its hinolinol form, where Z is not nucleophilic anion.

In an additional embodiment, the composition includes a compound having the formula II, and/or hinolinol form, where Z is not nucleophilic anion, and where the composition includes a cyclodextrin.

In various examples, the molar ratio of the compound to cyclodextrin is from about 10:1 to 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, for example, the molar ratio of the compound to cyclodextrin is about 1:2.

More preferably, the composition includes a compound having the formula

in which the cyclodextrin is β-cyclodextrin, more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In one embodiment, the OS is enforced pharmaceutical composition for oral maintenance.

In an additional embodiment, the pharmaceutical composition is formulated for parenteral administration.

In an additional aspect, the present invention relates to a dosage form containing or including stable compound comprising a lipophilic cation fragment associated connecting with the fragment of the molecule antioxidant, and anionic component for the specified cationic fragment, together with a pharmaceutically acceptable diluent and/or carrier and/or excipient, and

cationic fragment capable of enter into the mitochondria antioxidant molecules and

anionic component is not a halogen atom, and

anionic component is not nucleophilic and/or

anionic component does not show the reaction activity in relation to the cationic portion, the connecting fragment or molecule antioxidant.

In one embodiment, the molecule antioxidant is a quinone or China.

In other embodiments, the implementation of a molecule of an antioxidant selected from the group comprising vitamin E and derivatives of vitamin E, an antioxidant circuit, including mutilateral hydroxyanisol, mutilateral hydroxytrol total acceptors radicals, including derivateservlet fullerenes, spin traps, including the production is adnie 5,5-dimethylpyrrole-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

In one embodiment, a lipophilic cationic fragment represents a cation substituted or unsubstituted triphenylphosphane.

In one embodiment, the compound has a General formula I

and/or its hinolinol form, where the group R1, R2and R3that may be the same or different, selected from C1-C5alkyl (possibly substituted) groups or N, where n means an integer from about 2 to about 20, and where the symbol Z indicates directionspanel anion.

Preferably Z is selected from the group comprising alkyl - or arylsulfonate or nitrates.

Preferably each link carbon-carbon (C)n the bridge is full.

In an additional embodiment, the compound has the formula

and/or its hinolinol form, where Z is not nucleophilic anion.

In an additional embodiment, dosage form includes a compound having the formula II, and/or hinolinol form, where Z is not nucleophilic anion, and where the composition includes a cyclodextrin.

In various examples, the molar ratio of the compound to cyclodextrin is from about 10:1 to when is Erno 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, for example, the molar ratio of the compound to cyclodextrin is about 1:2.

More preferably the dosage form includes a compound having the formula

in which the cyclodextrin is β-cyclodextrin, more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In one embodiment, dosage form suitable suitable for oral administration.

In an additional embodiment, a pharmaceutical form suitable suitable for parenteral administration.

In an additional aspect of the present invention includes a dosage form suitable for oral administration containing as active ingredient the compound according to the present invention, the connection represents or prepared as a crystalline form and/or liquid form.

In an additional aspect, the present invention relates to a dosage form suitable for parenteral administration and which includes as an active ingredient the compound according to the present invention.

In an additional aspect, the present invention relates to a pharmaceutical composition and, suitable for the treatment of patients who may get relief from the reduction of oxidative stress or reduce the symptoms of aging, which contains or includes an effective amount of the compounds of the present invention in combination with one or more pharmaceutically acceptable excipients, carriers or diluents.

In one embodiment, the compound is a compound of formula I.

In one example, the compound forms a complex with cyclodextrin.

In various examples, the molar ratio of the 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, from about 2:1 to about 1:2 or about 1:1, for example, the molar ratio of the compound to cyclodextrin is about 1:2.

More preferably the compound is a compound of formula (III), and cyclodextrin is β-cyclodextrin, and more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In an additional aspect, the invention relates to a method of reducing oxidative stress in the cell, which involves the step of contacting the specified cell with the compound of the present invention.

In one embodiment, the compound is a compound of formula 1.

In one example, the compound forms to the complex with the cyclodextrin.

In various examples, the molar ratio of the 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, from about 2:1 to about 1:2 or about 1:1, for example, the molar ratio of the compound to cyclodextrin is about 1:2.

More preferably the compound is a compound of formula (III), and the cyclodextrin is β-cyclodextrin, and more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In one embodiment, the pharmaceutical composition is formulated for oral administration.

In an additional embodiment, the pharmaceutical composition is formulated for parenteral administration.

In an additional aspect, the present invention relates to pharmaceutical compositions suitable for the treatment of patients suffering from or predisposed to Parkinson's disease, Alzheimer's disease, horii's disease or hereditary ataxia ataxia, which contains or includes an effective amount of the compounds of the present invention in combination with one or more pharmaceutically acceptable excipients, carriers or diluents.

Preferably, said treatment is intended for patients suffering from or predisposed to a change in the state of frda.

In an additional aspect, the present invention relates to a method of treatment or prophylaxis of patients that could gain relief from reduced oxidative stress, which contains or includes a step of introducing a specified patient compounds of the present invention.

In one embodiment, the compound is a compound of formula I.

In one example, the compound forms a complex with cyclodextrin.

In various examples, the molar ratio of the 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, from about 2:1 to about 1:2 or about 1:1, for example, the molar ratio of the compound to cyclodextrin is about 1:2.

More preferably the compound is a compound of formula (III), and cyclodextrin allacts β-cyclodextrin, and more preferably the molar ratio of the compound to cyclodextrin is about 1:2.

In one embodiment, the specified introduction oral administration.

In another embodiment, the specified introduction parenteral administration.

In another aspect, the invention relates to a method of treatment or prophylaxis of patients who would feel relief from the reduction of oxidative stress or reduction of symptoms is torenia, which includes a step of introducing the patient connection of the present invention.

In another aspect, the invention relates to a method of treatment or prophylaxis of patients suffering from or predisposed to Parkinson's disease, Alzheimer's disease, horii's disease or hereditary ataxia ataxia, which contains or includes a step of introducing these patients compounds of the present invention.

Preferably, the method of treatment or prophylaxis relates to patients suffering from or predisposed to hereditary ataxia ataxia.

In another aspect, the invention relates to a method of reducing oxidative stress in cells, which includes a step of introducing into the cells of the compounds of the present invention.

In another aspect the invention relates to the use of compounds described above, upon receipt or production of drugs, dosage form or pharmaceutical composition effective for use in order to reduce oxidative stress in patients.

In another aspect the invention relates to the use of compounds described above, upon receipt or production of drugs, dosage form or pharmaceutical composition effective for use in order to reduce the symptoms of aging patients.

In an additional aspect, the invention Rel is referring to the use of compounds of the present invention for the production of medicines, dosage forms or pharmaceutical compositions effective for use in the treatment or prevention in a patient suffering from or predisposed to Parkinson's disease, Alzheimer's disease, horii's disease or hereditary ataxia ataxia, which contains or includes a step of introducing these patients compounds of the present invention.

Preferably the drug, dosage form or pharmaceutical composition is effective for use in the treatment or prophylaxis of patients suffering from or predisposed to ataxia of Pridraga.

In another aspect the invention relates to the use of compounds described above, upon receipt or production of drugs, dosage form or pharmaceutical composition effective for use in the reduction of oxidative stress in the cells.

Preferably specified receipt or production is carried out with the use of other material or materials, more preferably pharmaceutically acceptable diluents, excipients and/or carriers.

In an additional aspect, the present invention relates to a method of synthesis of compounds with a fragment or a fragment of the formula I

and/or its quinone form) where the group R1, R2and R3to the which may be the same or different, selected from C1-C5alkyl (possibly substituted) groups, where n means an integer from 2 to 20, this method contains or incorporates mixed with dextrin.

Preferably each link carbon-carbon (C)n the bridge is full.

In an additional aspect, the present invention relates to a method of synthesis of compounds having the formula

this method contains or includes blending cyclodextrin.

In an additional aspect, the present invention relates to a method of synthesis of compounds having the formula

essentially as described in the present description.

Any discussion of documents, acts, materials, devices, articles or the like, which is included in the present description of the invention made for the sole purpose of presenting the context of the present invention. Should not be considered a recognition that any or all of these positions, taken from a known level or which is common with known expertise in this area, important for the present invention as it existed before the priority date of each claim of this application.

Throughout this description, the word “include” or variations such as “comprises” or “comprising”should be understood as implying the criminal code of the related element, integer or stage, or group of elements, integers or steps, but not covering any other element, integer or stage, or group of elements, integers or steps.

Throughout the text of the description, the term “quinone”, used alone or in combination with another term to describe the oxidized form of the compounds should be understood as including within the scope of claims recovered form this connection, that is, hinolinol form. Similarly, the reference to the quinone, for example, when the structural description, also includes within the scope of the claims hinolinol form.

Throughout the text of the description, the term “China”, used alone or in combination with another term to describe the restored form of connection, it should be understood as including within the description of the oxidized form of this compound, that is, the form of the quinone. Similarly, the reference to China, for example, when the structural description, also includes within the scope of claims of the form of the quinone.

As used in the text of the description, the term “and/or” includes both meanings: “and”and “or” as options.

As used in the text of the description, the term “distribution coefficient and distribution coefficient (octanol : water)” refers to the distribution coefficient in the system Octan-1-ol/saline with FOSFA the tion buffer, specified at 25°C or 37°C (see Kelso, G.F., Porteous, C.M., Coulter, C.V., Hughes, G. Porteous, W.K., Ledgerwood, E.C., Smith, R.A.J. and Murphy, M.P. 2001, J. Biol. Chem., 276, 4588., Smith, R.A.J., Porteous, C.M., Coulter, C.V. and Murphy, M.P., 1999, Eu. J. Biochem., 263, 709. Smith, R.A.J., Porteous, C.M., Gane, A.M. and Murphy, M.P. 2003. Proc. Nat. Acad. Sci., 100, 5407), or the distribution coefficient in the system octanol/water, calculated using Advanced Chemistry Development (ACD) Software Solaris V4.67, as described in the publication Jauslin, M.L., Wirth, T., Meier, T., and Schoumacher, F., 2002, Hum. Mol. Genet. 11, 3055.

As used in the text, the phrase “acceptable for pharmaceutical drug” includes within its meaning not only the acceptability regarding pharmaceutical injection, but also in the composition, for example, acceptable stability, shelf life, hygroscopicity, cooking, etc.

As used in the text, “directionspanel anion represents an anion, which does not show the reaction activity as antioxidant molecules, lipophilic cation or the connecting portion. For example, if one such fragment connection includes a target of nucleophilic attack of the anion is not nucleophilic.

Though definitely in a wide scope, the invention is not limited to them, but also contains options for implementation, for which in the following description are examples.

In particular, a better understanding of the invention makes reference to Annex the proposed drawings.

Brief description of drawings

Figure 1 shows a diagram of penetration of amphiphilic antioxidant in the mitochondria, which shows the penetration of Mitogenome-C10 in a Horny mitochondrion.

Figure 2 shows the scheme for synthesis of a: Mitogenome-C3;: Mitogenome-C5; C: Mitogenome-C15.

Figure 3 shows the structure of the antioxidants number of Mitogenome and related compounds TRMR. The phospholipid shown in the same scale, aligned with antioxidants number of Mitogenome to denote the maximum possible depth of penetration originalnoy side chain in one piece phospholipid double layer. And: TRMR; IN: Makinon-C3;: Makinon-C5; D: Makinon-C10; S: Makinon-C15; F: phospholipid.

Figure 4 presents graphs showing the absorption and binding of antioxidants mitochondria, measured using ion-selective electrode. A: Makinon-C3;: Makinon-C5; C: Makinon-C10; D: Makinon-C15. The left panel presents the mitochondria (1 mg protein/ml) in the presence of rotenone, and then add antioxidants five successive additions of 1 μm (black arrows at the top) to calibrate the response of the electrode. Right panel electrodes were first calibrated five successive additions of 1 μm (black arrows at the top)and then added the mitochondria (1 mg protein/ml). In all cases, what was b added succinate to generate a membrane potential, and FCCP was added to its dissipation. Data represent typical responses to the experiment repeated at least 2-3 times.

Figure 5 presents graphs showing the antioxidant efficiency of the antioxidant. A: Mitochondria were initiated by succinate (black arrows) or by incubation with ATP regenerating system, including ATP, pyruvate of phosphoenol and pyruvate kinase (white arrows). After 30 sec pre-incubation with various analogues of Mitogenome, TRMR or media induced oxidative stress by adding 50 μm FeCl2and 300 μm N2About2. After 15 minutes incubation at 37°C set the lipid peroxidizable measurement TBAR. Results are mean ± interval of two independent experiments. Lightweight protective effect of Mitogenome C-5 from lipid peroxidation in the presence of ATP caused a certain amount of Mitogenome-C5 dobavlennogo from the core solution and in originalnoy form. In: Mitochondrial membrane potential induced by succinate or ATP regenerating system, measured by the accumulation of [3H]TPMP. Results are mean ± interval two definitions 25-minute incubation. Membrane potentials after 5 minutes of incubation were the same (data not shown). With: Measured concentration dependence prevent the oil accumulation TBAR antioxidants. All incubation was performed in the presence of succinate as described for A. the Results are expressed as % inhibition of TBARS composition, taking the value of sample subjected to FeCl2/H2O2in the absence of analogues of Mitogenome for 0% inhibition and the control sample (without adding FeCl2/H2O2) for 100%. Data shown represent typical titration results for each concentration, some three times ± SD. IC50concentration to prevent lipid peroxidation. The results indicate the mean values ± sem calculated from three independent titrations, as shown in paragraph C. Statistical significance relative to the IC50for mitogenome-C3 was determined using training test on the two sloping sections: *p<0,05; **p<0,005.

Figure 6 presents a graph showing the effect of mitogenome-C10 and Mitogenome-C3 in coronary sinus flow.

Figure 7 presents a graph showing the effect of Mitogenome-C10 and Mitogenome-C3 on diastolic pressure in the left ventricle.

On Fig presents a graph showing the effect of Mitogenome-C10 and Mitogenome-C3 cardiac rhythm.

Figure 9 presents a graph showing the change of rhythm in the left ventricle.

Figure 10 presents a graph showing the effect of Mitogenome-C10 and Mitogenesis on postischemic mitochondrial respiratory function.

Figure 11 presents graphs illustrating the stability of pure Mitogenome-C10 (series No. 3) in clean glass vials at 40°C, 75% RH; 25°C, 50% relative humidity; and 5°C over silica gel.

On Fig presents a bar graph showing the stability of Mitogenome-C10 series (No. 4) at 25°C, 50% relative humidity.

On Fig presents a bar graph showing the stability of the complex Mitogenome-C10 and β-cyclodextrin (1:1) at 4°C over silica gel, 50% relative humidity and 40°C, 75% relative humidity.

On Fig presents a bar graph showing the stability of the complex Mitogenome-C10 and β-cyclodextrin (1:2) at 4°C over silica gel, 25°C, 50% relative humidity and 40°C, 75% relative humidity.

On Fig presents a bar graph showing the stability of the complex Mitogenome-C10 and β-cyclodextrin (1:4) at 4°C over silica gel, 25°C, 50% relative humidity and 40°C, 75% relative humidity.

On Fig presents a graph showing the stability of nelfinavir Mitogenome-C10 in the water.

On Fig presents a graph showing the stability of nelfinavir Mitogenome-C10 in 0.01 M HCl.

On Fig presents a graph showing the stability of nelfinavir Mitogenome-C10 in IPB, pH 7.4.

On Fig presents a graph showing the stability of nelfinavir Mitogenome-C10 50% MeOH.

On Fig presents a graph p is practice showing stability in the solid state nelfinavir of Mitogenome-C10 at 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) in water.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) in 0.01 M HCl.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) in IPB, pH 7.4.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) 50% of the Meon.

On Fig shows a graph showing the stability in solid state complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) at 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) in water.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:1) in 0.01 M HCl.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:1) in IPB, pH 7.4.

On Fig presents a graph showing the stability of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:1) in 50% methanol.

On Fig p is edstaven graph showing stability in solid state complex mesilate of Mitogenome-C10-β-cyclodextrin (1:1) at 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel.

On Fig shows graphs of changes in the concentration of Mitogenome-C10 in plasma after a single IV (A) (10 mg/kg) and oral (B) (50 mg/kg) administration to rats of nelfinavir Mitogenome-C10 in the form of a complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) (n=5). Pharmacokinetic parameters derived from these data, presented in table 11.

Detailed description of the invention

As noted above, the main objective of the present invention is directed introduction of compounds in mitochondria, mainly for the treatment and/or prevention with the reduction of oxidative stress.

Mitochondria have significant membrane potential, which is estimated to be 180 mV for its internal membrane (negative inside). Due to this potential, membrane permeable lipophilic cations are accumulated in several hundred layers inside the mitochondrial matrix.

The inventors have found that when connecting lipophilic cations (e.g., lipophilic cation triphenylphosphane) with one molecule of antioxidant formed of amphiphilic compound may be introduced into the mitochondrial matrix is in intact cells. Antioxidant then directed enters the main place of formation of free radicals and reactive fragments of oxygen inside the cell, and is not distributed statistically.

The authors additionally found that the properties of an antioxidant, such as, for example, the nature of the antioxidant molecules, physical and chemical properties of the connecting fragment, such as, for example, the length or the lipophilicity of the connecting portion and/or the nature of the lipophilic cation affect the efficiency of the antioxidant in vivo and enhances the functional activity of antioxidant compounds. For the antioxidant compounds of the present invention efficacy in vivo may in part include appropriate bioavailability suitable stability, suitable pharmacokinetics, a suitable antioxidant activity and/or appropriate mitochondrial target and/or accumulation.

In principle, any lipophilic cation and any antioxidant, capable of transporting in and/or through the mitochondrial membrane and accumulated on or within the mitochondria of intact cells, can be used to obtain compounds of the present invention.

However, preferably, the lipophilic cation represented cation triphenylphosphine presented in this description as an example. The other is their lipophilic cations, which can be covalently linked with antioxidants in accordance with the present invention include cations of tribenzylamine and phosphonium. In some examples, the antioxidant compounds of the present invention the lipophilic cation is connected with one molecule of antioxidant rich linear carbon chain containing from 1 to about 30 carbon atoms, for example, from 2 to about 20, 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 of a linear carbon chain contains 10 carbon atoms.

Preferably the carbon chain is alkylenes group (for example, C1-C20or1-C15), although the carbon chain, which optionally comprise one or more double or triple links, also included in the scope of claims of the present invention. It also includes a carbon chain containing one or more substituents such as hydroxyl, carboxyl or amide groups and/or contain one or more side chains or branches, such as those selected from unsubstituted or substituted alkyl, alkenyl or etkinlik groups. It also includes a carbon chain, which include more than about 30 carbon atoms, but whose length is equivalent to Lin who Inoi saturated carbon chain, containing from about 1 to about 30 carbon atoms.

Specialists in this field will appreciate the fact that the fragments that differ from the linear alkilinity, can be used for covalent compounds, the antioxidant molecules with lipophilic cation, for example, substituted or branched alkyl group of the peptide bond, etc.

In some embodiments, the implementation of the lipophilic cation is connected with one molecule of antioxidant linear chain alkalinous group containing from 1 to 10 carbon atoms, such as, for example, ethylene, propylene, butylene, Panteleeva or dellanave group.

Molecules of antioxidant used in the present invention include those that require interaction with the reducing agents for the manifestation of antioxidant activity, regardless of whether it is the original activity of antioxidant or circulating antioxidant activity or both types of activity. For example, the antioxidant compounds of the present invention, which includes as an active antioxidant molecules hinolinol molecule can be introduced in the form of a quinone. To act as an antioxidant, that is, to possess antioxidant activity, quinone must be restored to chinoline forms of interaction with a reducing agent such as, e.g. the measures mitochondrial reducing agent, such as Complex II, for the manifestation of the source of antioxidant activity. The subsequent interaction of the oxidized quinone form by reducing agents can lead to the manifestation of the circulation antioxidative activity.

Other examples of molecules of antioxidants that can be used in the present invention include those that already exist in the restored form and does not require interaction with the reducing agents for the manifestation of the source of antioxidant activity. Despite this, the subsequent interaction of the oxidized form of such molecules with antioxidant mitochondrial reducing agents can lead to the manifestation of circulating antioxidant activity. For example, the antioxidant molecule of vitamin E can be entered in restored form, and therefore does not require interaction with the reducing agents for the manifestation of the source of antioxidant activity, but may then interact with reducing agents, such as, for example, endogenous quinone Fund, with circulation in the result of antioxidant activity.

Other examples of molecules of antioxidants that can be used in the present invention include those that do not give the circulation activity when interacting with mitochondrial what ostanovitesj.

Examples of molecules of antioxidants used in the present invention include vitamin E and derivatives of vitamin E, antioxidants circuit, such as mutilateral hydroxyanisol, mutilateral hydroxytoluene, hinely and total acceptors radicals, such as derivationally fullerenes. In addition, can be used spin traps, which interact with free radicals with the formation of stable free radicals. They include derivatives of 5,5-dimethylpyrrole-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

The preferred antioxidant compounds, including anti-oxidant compounds of General formula I and II, can be easily obtained, for example, by the following reaction:

The General scheme of the synthesis consists in heating a precursor containing a suitable tsepliaeva group, preferably alkylsulfonyl, bromine - or iodine-containing precursor, with more than 1 equivalent of triphenylphosphine, under argon atmosphere for several days. Postname connection is then isolated in the form of its salts. To do this, the product is triturated repeatedly with diethyl ether until then, until there is a whitish solid. It is then dissolved in chloroform and the and dichloromethane and precipitated in diethyl ether to remove excess triphenylphosphine. This is repeated up until the solid is no longer dissolved in chloroform. At this point, the product is recrystallized several times from a suitable solvent, such as chloroform, acetone, ethyl acetate, or higher alcohols.

The preferred method of synthesis, which can be used to obtain a stable preferred form, is directed insertion in mitochondria antioxidant compounds of formula III (also referred to in this description mesilate of Mitogenome-C10 or methanesulfonate of Mitogenome C-10), described below in example 1.

Also it will be clear that the anion antioxidant compounds, thus obtained, can be easily exchanged for another pharmaceutically or pharmacologically acceptable anion, if this is desirable or necessary, when using ion exchange or other technologies known in this field.

Applicants have found that the stability of the salt form of antioxidant compounds increases when the anion does not show the reaction activity as antioxidant molecules, the connecting piece or a lipophilic cationic fragment. For example, in the case of the preferred examples of the antioxidant compounds of the present invention, the anion is not nucleophilic. It is also desirable that the anion was a pharmaceutically acceptable Academy of Sciences is one. Also preferably, for the pharmacological composition of the anion showed no reaction activity in relation to other agents within the formula of composition.

Examples of non-nucleophilic anions include hexafluoroantimonate, arsenate or phosphate, or tetraphenylborate, Tetra(perftoralkil)borate or other tetrafluoroborate, triftorbyenzola, aryl - or alkyl sulphonates, such as methanesulfonate and p-toluensulfonate, and phosphates.

Examples of pharmaceutically acceptable anions include halogen ions such as fluorine ion, chlorine ion, bromine ion and iodine ion, anions of salts of inorganic acids, such as nitrate, perchlorate, sulfate, phosphate and carbonate; pharmaceutically acceptable anion salts of the lower alkylsulfonic acids, such as methanesulfonic acid, and salts ethanesulfonic acid; farmatsevticheskii acceptable anion salts arylsulfonic acids, such as salts of benzosulfimide acid, 2-naphtalenesulfonic acid and p-toluensulfonate acid; pharmaceutically acceptable anion salts of organic acids, such as salts of trichloroacetic acid, triperoxonane acid, hydroxyoctanoic acid, benzoic acid, almond acid, butyric acid, propionic acid, formic acid, fumaric acid, succinic acid, citric acid, tartaric acid, Savelev the th acid, maleic acid, acetic acid, malic acid, lactic acid and ascorbic acid; and pharmaceutically acceptable anion salts of acidic amino acids, such as salts of glutamic acid and aspartic acid.

In the case of the preferred examples of the antioxidant compounds of the present invention, the precursor anion of halogen substituted in the aryl and alkylsulfonate anions. Examples include, but not limit the scope of the claims, bansilalpet, p-toluensulfonate, 2-naphthalenesulfonate, methanesulfonate, ethanesulfonate, propanesulfonate. Especially preferred anion is the anion of methansulfonate. As described above, an example of antioxidant compounds according izobreteniya, in which the anion is methanesulfonate, is particularly preferred antioxidant compound of the formula III, called the description text of the methanesulfonate of Mitogenome-C10 or mesilate of Mitogenome-C10.

The same General technique can be used to produce a wide range of targeted insertion in mitochondria connections fragment R of different antioxidants that are attached to the fragment or fragments triphenylphosphane (or other lipophilic cationic fragment). They will include a series of derivatives of vitamin E, in which the length of the bridge linking functional group VI is Amin E fragment triphenylphosphane (or other lipophilic cationic fragment), is different. Other antioxidants that may be used as group R include antioxidants circuit, such as mutilateral hydroxyanisol, mutilateral hydroxytoluene, hinely and total acceptors radicals, such as derivationally fullerenes. In addition, spin traps, which interact with free radicals with the formation of stable free radicals, can also be synthesized. They will include derivatives of 5,5-dimethylpyrrole-N-oxide, tert-butylnitrite, tert-nitrobenzoyl, α-phenyl-tert-butylnitrone and related compounds.

It will be clear that the antioxidant compounds of the present invention, as for any drug activity in vitro in no way is the determining parameter for the functionality or effectiveness in vivo. Antioxidant activity the antioxidant compounds of the present invention can be determined by methods such as those described in this text, using, for example, isolated mitochondria and/or the selected cells. Although it is indeed true that in order to be used as directed insertion in mitochondria antioxidant compounds of the present invention, the antioxidant compound should be appropriately high antioxidant dei is a journey in such a sample, but to be effective in vivo, targeted insertion in mitochondria antioxidant compound must exhibit other desirable physical and chemical properties, for example, to have a suitable bioavailability, stability or oxidation efficiency.

Examples of the antioxidant compounds that show good antioxidant activity and also show poor bioavailability relative to the Department's target in vivo, include Coenzyme Q (CoQ) and Idebenone. Both of these compounds should be introduced at a very high dosage (for example, 0.5-1.2 g) to achieve minimal clinical effects in patients with people.

Examples of directionally injected into the mitochondria antioxidant compounds of the present invention exhibit good antioxidant activity and bioavailability and are therefore effective in vivo at low doses. Determination of the bioavailability of the preferred amphiphilic, directed insertion in mitochondria antioxidant compounds of the present invention, nelfinavir of Mitogenome-C10 and its complex with a cyclodextrin, presented in the description in example 11. The authors suggest that the antioxidant compounds of the present invention are effective in the direction of introduction into the mitochondria antioxidant activity, displaying one or more mainly what usest, based on the availability in the crystalline or solid form and use for the preparation of compounds in solid form, have increased stability, improved bioavailability and/or improved anti-oxidation effect. Physical and chemical properties of the antioxidant compounds of the present invention, as the authors suggest, is not tied to any theory, belong to the preferred properties of the antioxidant compounds of the present invention, and allow them to be used in the compositions, combinations and methods in those applications in which the antioxidant compounds known level may be less suitable because of their chemical and physical properties.

In some embodiments the invention, the antioxidant compound is chinoline derivative of the formula II as defined above. For example, chinoline derivative according to the invention is a compound of Makinon-C10 (specific salt which is a compound of formula (III)as defined above. Another example of compounds of the present invention is a compound of formula I, in which (C)nrepresents (CH2)5and hinely fragment is the same as Mitogenome-C10 mentioned in the description Makinana-C5 (see figs). Another Supplement is inim example compounds of the present invention is a compound of formula I, in which (C)nrepresents (CH2)3, hinely fragment is the same as Mitogenome-C10, which is called in the description Makinana-C3 (see figv). Another example of compounds of the present invention is a compound of formula I, in which (C)nrepresents (CH2)15and hinely fragment is the same as Mitogenome-C10 mentioned in the description Makinana-C15 (see figv).

Once received, the antioxidant compound of the present invention in any pharmaceutically suitable form and may include pharmaceutically acceptable carriers, fillers, solvents, complexing agents or additives, is injected into a patient in need of treatment and/or prevention. After the introduction of the connection will be directed to create antioxidant activity in the mitochondria inside the cells of the patient.

Antioxidant compounds of the present invention can be administered to the patient orally and/or parenterally.

Antioxidant compound must be made in the form of a stable, safe pharmaceutical composition for administration to a patient. The composition may be obtained according to known methods by dissolving or suspendirovanie amount of antioxidant ingredient in a diluent. The number of antioxidant compounds accounted for the ing from 0.1 mg to 1000 mg per ml of diluent. Can be added acetate, phosphate, citrate or glutamate buffer to bring the pH of the final composition to a value of from 5.0 to 9; may carbohydrates controller isotonicity or regulator isotonicity on the basis of a polyhydric alcohol and a preservative selected from the group comprising m-cresol, benzyl alcohol, methyl, ethyl, propyl and butylparaben, and can also be added to the phenol. To obtain the desired concentration of the solution using a sufficient quantity of water for injection. If desired, the composition may include additional controllers isotonicity such as sodium chloride, as well as other fillers. However, such fillers must support the overall toychest antioxidant compounds.

The term buffer, buffer solution or a solution with a buffer when used with respect to the hydrogen ion concentration or pH, refers to the ability of systems, especially aqueous solution to resist change in pH upon addition of acid or base, or upon dilution with a solvent. A distinctive feature of the solutions with buffers that are undergoing a slight change of pH by adding acid or base, is the presence of either a weak acid and a salt of a weak acid or weak base and the salt of the weak base. An example of the first mentioned system is we are acetic acid and sodium acetate. The change in pH slightly, as the number of hydroxyl ions does not exceed the buffer capacity of the system to neutralize them.

Stability of parenteral composition of the present invention is increased while maintaining the pH of the composition in the range of approximately from 5.0 to 9.5. Other intervals of pH include, for example, from 5.5 to 9.0, or from 6.0 to 8.5, or from about 6.5 to 8.0, or from 7.0 to 7.5.

The buffer used in the practical implementation of the present invention, any selected from among the following, for example, acetate buffer, phosphate buffer or glutamate buffer, the preferred buffer is phosphate buffer.

Fillers or carriers can also be used to accelerate the introduction of a connection. 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.

The stabilizer may be included in the composition of the present invention, but, importantly, not necessarily. However, if it is included in the formulation of the composition, the stabilizer used in the practical implementation of the invention is a carbohydrate or polyhydric alcohol. Polyhydric alcohols include compounds such as sorbitol, Mann is t, glycerin and polyethylene glycol (Page). Carbohydrates include, for example, mannose, ribose, trehalose, maltose, Inositol, lactose, galactose, arabinose, lactose-free.

Suitable stabilizers include, for example, polyhydric alcohols such as sorbitol, mannitol, Inositol, glycerol, xylitol and copolymer polypropylene/ethylene glycol, and various glycols (Page) molecular weight 200, 400, 1450, 3350, 4000, 6000 and 8000).

The United States Pharmacopeia (USP) argues that the drugs contained in the containers for multiple doses should be added antimicrobial agents in bacteriostatic or fungistatic concentrations. They must be present in sufficient concentration at the point of use to prevent the growth of microorganisms, inevitably falling into the drug when removing part of the contents of the hypodermic needle and syringe, or using other invasive tools for delivery, such as pen injectors. Antimicrobial agents should be evaluated for compatibility with all other components of the formulation, and their activity should be evaluated in the General formula, to make sure that the specific agent that is effective in one recipe, is not effective in another. Do not rarely be able to detect that a particular agent will be effective in one recipe, but not effective in other Rotz is round.

Preservative, in General, the pharmaceutical sense, is a substance that prevents or inhibits the growth of microbes and can be added to the pharmaceutical composition for this purpose, to avoid further damage to the structure of microorganisms. Although the amount of preservative is small, however, it can affect the overall stability of the antioxidant compounds. Thus, even the choice of preservative may be difficult.

Although the preservative for use in the practice of the present invention may be 0.005 to 1.0% (m/o), the preferred interval for each of a preservative, one or in combination with others, is: benzyl alcohol (0.1 to 1.0%), or m-cresol (0.1 to 0.6%), or phenol (0,1-0,8%),or a combination of methyl- (0,05-0,25%), ethyl-, propyl - or butyl (0,005%-0,03%) parabens. Parabens are complex lower alkalemia esters of para-hydroxybenzoic acid.

A detailed description of each preservative presented in Remington''s Pharmaceutical Science”, as well as Pharmaceutical Dosage Forms: Parenteral Medications, Vol.1, 1992, Avis et al. For these purposes crystalline chloromethane salt trientine can be entered parenterale (including subcutaneous injections, intravenous, intramuscular, intradermal injection or infusny methods of injection or inhalation sputtered medicinal substances in the composition of the dosage form containing the traditional non-toxic, pharmaceutically acceptable carriers, auxiliaries and fillers.

Also, it may be desirable to add sodium chloride or other salt to regulate toychest pharmaceutical composition, depending on the selected controller isotonicity. However, this does not necessarily depend on the specific composition. Parenteral formulations must be isotonic or substantially isotonic, otherwise there is a strong irritation or have any pain at the injection site.

Desirable isotonicity can be achieved through the use of sodium chloride or other pharmaceutically acceptable agents such as dextrose, boric acid, sodium tartrate, propylene glycol, polyols (such as lures or sorbitol) or other inorganic or organic soluble substances. Typically, the composition is isotonic with the blood of the patient.

If desired, the parenteral composition can be thickened with a thickener such as methyl cellulose. The composition can be prepared in the form of emulsions of the type water-in-oil or oil-in-water. Can be used any of a wide variety of pharmaceutically acceptable emulsifying agents, including, for example, powder Arabian gum, nonionic surfactant or an ionic surfactant.

Can have what I also want to add in a pharmaceutical composition suitable dispersant or suspendisse agent, these may include, for example, aqueous suspensions, such as synthetic and natural gums, i.e. tragakant, Arabian gum, alginate, dextran, carboxymethylcellulose sodium, methylcellulose, polyvinylpyrrolidone or gelatin.

Filling the very large values for pharmaceutical products is water. Water suitable for parenteral administration should be obtained either by distillation or reverse osmosis. Only such means it is possible to separate different liquid, gas and solid contaminant from the water. Water for injection is the preferred aqueous media for use in the pharmaceutical compositions of the present invention. Water can be purged with gaseous nitrogen to remove any traces of oxygen or free radicals of oxygen from the water.

It is also possible that other ingredients were present in parenteral pharmaceutical compositions of the present invention. Such additional ingredients may include wetting agents, oils (for example, vegetable oil, like sesame oil, peanut oil or olive oil), analgesics, emulsifiers, antioxidants, fillers, regulators isotonicity, metal ions, oily fillers, proteins (e.g. human serum albumin, gelatin or proteins) and Zvi who cerioni (for example, such amino acids as betaine, taurine, arginine, glycine, lysine and histidine). Such additional ingredients, of course, must not adversely affect the overall stability of the pharmaceutical composition of the present invention.

Containers are also a part of the composition for injection and can be considered as a component, as there is no container, which would have been completely insoluble or not was influenced to some extent by the liquid which it contains, especially if the liquid is water. Therefore, the choice of container for a specific injection should be based taking into account the composition in the container, as well as solution and treatment for which it is intended.

In order to ensure the introduction of a needle from a hypodermic syringe into the vial with multiple dose and re-sealing after removal of the needle, each vial is closed with a rubber stopper attached to the place of the aluminum plate.

Caps for glass bubbles, such as West 4416/50, 4416/50 (with Teflon top) and 4406/40, Abbot 5139, or any equivalent tube can be used for closing the bubbles with a dosage form of the drug. These caps are tested on the integrity of the cover when tested using the scheme of treatment of the patient, for example, the cover must withstand at least about 100 injections.

Each is th component of the pharmaceutical composition, as described above, known in the field and described in the book of Pharmaceutical Dosage Forms: Parenteral Medications, Vol.1, 2nded., Avis et al., Mercel Dekker, New York, N.Y. 1992, which is incorporated into this description by reference in its entirety.

The process of obtaining the above composition includes stage compounding, sterile filtration and filling. The technique of compounding, for example, to enable the dissolution of ingredients in a specific order, for example, first, a preservative, then the stabilizer/regulator isotonicity, buffers, and then antioxidant compound, or the dissolution of all ingredients, forming a parenteral composition, at the same time. An example of one way of receiving parenteral composition for injection is the dissolution of antioxidant compounds, for example, complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2)in water and diluting the resulting mixture in a physiological solution with phosphate buffer.

In an alternative embodiment, the parenteral compositions of the present invention is produced by mixing ingredients according to standard techniques. For example, the selected components may be mixed in the mixer, or other standard device to produce a concentrated mixture, which can then be brought to the final concentration and viscosity by the addition of water, a thickening agent, a buffer, 5%-gallumine human serum or more soluble substances to regulate isotonicity.

Alternatively, the antioxidant compound can be packaged as a dry substance and/or powder, the recovered solvent to form a parenteral composition according to the present invention for use in the time of recovery.

In addition, the preparation process may include any suitable sterilization method in the development of parenteral composition of the present invention. Typical sterilization processes include filtration, process steam (moist heat), dry heat, gases (e.g., ethylene oxide, formaldehyde, chlorine dioxide, propylene oxide, beta-propiolactone, ozone, chlorpicrin, peracetic acid, hydrogen bromide and the like), radiation exposure and aseptic handling.

Suitable route of parenteral administration include intravenous, intramuscular, subcutaneous, intradermal, subdermal, intra-articular, intrathecal, intraperitoneal, etc. introduction. Intravenous route of administration is preferred. Introduction through the mucous membrane is also valid. The dosage and dosage regimen will depend on the weight and health of the patient.

Pharmaceutically acceptable excipients, carriers, diluents, complexing agents or additives may be selected, for example, to increase the stability of the antioxidant is to be placed, acceleration of synthesis or composition of the pharmaceutical composition and/or to increase the bioavailability of the antioxidant compounds.

For example, the molecules of the medium, such as cyclodextrin and its derivatives, are well known in this area because of their potential to act as complexing agents that can change the physico-chemical properties of drug molecules. For example, cyclodextrins can stabilize (as from thermal effects, and oxidation effects), to reduce volatility and to change the solubility of active agents with whom they are connected in the complexes. Cyclodextrins are cyclic molecules consisting of an annular links glucopyranose, which form a toroidal structure. The inner part of the molecule of cyclodextrin is hydrophobic, while the external part is hydrophilic, thus ensuring the solubility of the cyclodextrin molecules. The degree of solubility can be changed by substitution of hydroxyl groups on the outside of the cyclodextrin. Similarly, the hydrophobicity of the inner part can be modified by substitution, although it is generally hydrophobic nature of the inner part allows to accumulate relatively hydrophobic molecules guests inside the cavity. The accumulation of one molecule inside the other izvestniak complexation, and the resulting product is called the adduct. Examples of cyclodextrin derivatives include sulfosalicylate, multiselection, hydroxypropylcellulose and their salts.

Methods of obtaining pharmaceutically acceptable composition comprising the adduct directionally injected into the mitochondria antioxidant compounds, in this case Mitogenome-C10 in complex with β-cyclodextrin, disclosed in the present description in example 1 and example 7. Methods of obtaining pharmaceutically acceptable compositions containing the preferred adduct is directed insertion in mitochondria antioxidant compounds nelfinavir of Mitogenome-C10 in complex with β-cyclodextrin, disclosed in the present description in example 9 and example 10.

Physico-chemical properties, including, for example, the pharmaceutical properties of complex antioxidant compound-cyclodextrin, can be modified, for example, by changing the molar ratio of the antioxidant compound to cyclodextrin or change him. For example, for the preferred antioxidant compounds of General formula I to the molar ratio of the antioxidant compound to cyclodextrin (antioxidant compound:cyclodextrin) may 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:ili 1:1. In another example, the preferred molar ratio of the explanatory antioxidant compounds Mitogenome-C10 to cyclodextrin is 1:2, and the cyclodextrin is β-cyclodextrin.

Alternatively, pharmaceutically suitable form of antioxidant compounds can be obtained, so as to increase the stability and bioavailability of antioxidant compounds. For example, tablets can be coated with enteric coatings prevent release of antioxidant compounds in the stomach, or to reduce the risk of unpleasant side effects or maintain the stability of the antioxidant compounds, which otherwise may be subject to destruction under the influence of the gastric environment. Most polymers used for this circuit are a polyacid, which act by virtue of the fact that their solubility in water depends on pH and demanding conditions with a pH higher than normal values existing in the stomach.

One preferred type of oral structures with the regulated allocation is enteric coated solid dosage forms. Enteric coatings facilitate the introduction of compounds that are physically remaining in the dosage form over a certain period of time when exposed to gastric juice, while Ki is Ichnya coating designed to dissolve in the intestinal fluid and be ready to absorb. Delay absorption depends on the rate of flow through the gastrointestinal tract, and therefore the rate of release of the stomach is an important factor. For some routes of administration dosage form multiple types, such as granules, it may be preferable to disposable type. Therefore, in one embodiment of the invention antioxidant compounds according to the invention may be contained in the dosage form multiple type with enteric coating. In a more preferred embodiment, the medicinal form of antioxidant compounds are prepared by obtaining particles containing antioxidant compound agent of enteric coating on the inert core material. These granules can lead to prolonged absorption of antioxidant compounds with good bioavailability.

Typical agents of enteric coatings include, but not limit the scope of the claims, the phthalate of hydroxypropylmethylcellulose, the copolymer of methacrylic acid and a complex of methacrylic acid, polyvinyl acetate-phthalate and acetate-phthalate cellulose.

The preferred examples of the antioxidant compounds of the present invention and/or compositions and/or complexes are predominant pharmaceutical properties. For example, one of them easy to prepare the compositions, they are chemically and physically stable, is egco soluble in water, have low hygroscopicity and have good shelf life.

Hereinafter the invention will be described in more detail with reference to the following, not limiting the scope of the claims of the experimental part.

Example 1. Synthesis of Mitogenome-C10

The following describes a preferred method for the synthesis of preferred stable salt explanatory directed insertion in mitochondria antioxidant compounds Mitogenome-C10, nelfinavir of Mitogenome-C10 and its complex with the cyclodextrin.

Stage 1

Scheme:

Stage:

1. Idebenone (A1, 0.25 kg of 0.74 mol) is dissolved in 2.5 l DHM chemically pure and the mixture is then cooled to 10±3°C in inert atmosphere.

2. The triethylamine (0,152 kg, 1.5 mol) is added in one portion at ambient temperature and the mixture advocate to equilibrium at 10±3°C.

3. Then slowly add a solution of methanesulfonamide (0.094 kg, 0.82 mol) in 0.5 l DHM such a rate as to maintain an internal temperature of about 10-15°C. (Under these conditions, adding expires after 75 minutes).

4. The reaction mixture was stirred additionally for 15-30 minutes.

5. IPC checked for completion by TLC (Rf0,65, 5% ethanol/dichloromethane).

6. The mixture is then washed with water (0,85 l) and saturated aqueous solution b is sodium carbonate (0,85 l).

7. The organic layer is evaporated to obtain the red liquid under reduced pressure at 40-45°C. After drying for an additional 2-4 hours under high vacuum at ambient temperature the resulting crude A2 are used directly in the next stage. The output is not known, as the fluid captures the solvent.

Stage 2

Scheme:

Stage:

1. Mesilate idebenone (A2, provided 100% of the output from the last stage, 0.31 kg, of 0.74 mol) is dissolved in 2 l of methanol and the mixture is then cooled to 0-5°C under inert atmosphere.

2. Parts add sodium borohydride (0.03 kg, of 0.79 mol) with such speed that the internal temperature did not exceed 15°C. Completion of the reaction will be accompanied by a color change: red→yellow (In these conditions, the addition is completed in 20 minutes).

3. The reaction mixture was stirred additionally for 10-30 minutes.

4. IPC checked for completion by TLC (A3 Rf0,60, 5% ethanol/dichloromethane, A2 Rf0,65).

5. Then the reaction being removed by adding 2 l of a 2M solution of hydrochloric acid and three times extracted with 1.2 l of dichloromethane.

6. The combined organic phases are then washed once with 1.2 l of water and dried over anhydrous magnesium sulfate (0.24 kg).

7. Then the organic phase is evaporated and onigen the m pressure get syrup yellow-brown color at a temperature of 40-45°C. After drying for an additional 2-8 hours under high vacuum at ambient temperature the resulting crude product A3, 0,304 kg, 98% yield, are used directly in the next stage.

Stage 3

Scheme:

Stage:

1. Pieces of triphenylphosphine (0,383 kg of 1.46 mol) are added to the mesilate of idebenone (A3, 0,304 kg, 0.73 mol) of the appropriate size round bottom flask.

2. Then the flask attached to a rotary evaporator and its contents are heated under vacuum to a temperature bath at 80-85°C.

3. At this temperature, the mixture should form a homogeneous melt. As soon as the melt is formed, the generation of gas is not observed, replace vacuum inert atmosphere and the mixture is gently rotate in a bath whose temperature is set at 80-85°C for approximately 3 days.

4. IPC checked for completeness methods1H and31P NMR. Requires a minimum conversion of 95% for further processing of the product.

5. Then the mixture is cooled approximately to room temperature and dissolved in 0.8 l of dichloromethane.

6. Then add parts to 3.2 l of ethyl acetate at low heat until precipitation of the desired product from the excess triphenylphosphine.

7. A small amount of solvent is removed by evaporation under reduced pressure (to remove DHM) and the remaining with the offer then cooled to about ambient temperature and decanted.

8. The remaining sticky residue is then subjected to the same washing procedure twice more, and then finally dried under high vacuum to constant weight and get a yellow-brown foam product 0,441 kg, 89% yield (note: the product still contains some solvent, see NMR). Thus obtained A4 are used directly in the next stage.

Stage 4

Scheme:

Stage:

1. Crude salt nelfinavir methanol (0,44 kg, about to 0.65 mol) is dissolved in 6 l of anhydrous DHM and the flask is rinsed with oxygen.

2. The contents of the flask intensively stirred under oxygen atmosphere for 30 minutes to ensure saturation of the solvent gas.

3. 0.1 l of a solution of 0.65 M NO2in a dry DMH (2 mol % NO2) just add one portion and the mixture is intensively stirred under oxygen atmosphere for 4-8 hours at ambient temperature.

4. Then spend IPC check on the degree of completion (method1H NMR and possible31P NMR).

5. If the oxidation is not complete, then add another 2 mol % of NO2in the form of a solution in DHM. This should bring the reaction to completion. IPC is performed as described above. In these conditions for completion of the reaction requires 8 mol% of NO2in the form of a solution in DHM.

6. Then dissolve the al is removed by evaporation under reduced pressure and get syrupy residue of red. This residue is dissolved in 2 l of dichloromethane at 40-45°C.

7. Then add parts to 3.2 l of ethyl acetate at low heat until precipitation of the desired product. A small amount of solvent is removed by evaporation under reduced pressure (to remove DHM) and the remaining mixture was then cooled to about ambient temperature and decanted.

8. Oily residue is finally dried under high vacuum to constant weight and get glassy product red (419 g, 94% yield). Thus obtained A5 are used directly in the next stage.

Stage 5

Scheme:

Stage:

1. Crude salt nelfinavir of mitogenome (A5, 0,419 kg) dissolved in 6 l of water at low heat at 40-43°C.

2. Beta-cyclodextrin, 1.24 kg, separately dissolved in 20 liters of water by heating at 60°C.

3. These two solution is cooled approximately to room temperature and combine with obtaining a homogeneous mixture. This solution should be stored at a temperature of <5°C.

4. Then this solution orange frozen at -20°C and lyophilizers portions to constant weight (at least 48 hours).

5. The resulting solid is then gently crushed with obtaining homogeneous free flowing powder yellow-orange (1,433 kg).

Conducted Alta is native synthesis, at which stage of oxidation 3 stage 4 the synthesis method described above is achieved by bubbling oxygen through the solution, which indicates the possibility of oxidation reactions on the creature until end of oxidizing means, different from the oxidation of NO2.

Example 2. Synthesis directed insertion in mitochondria antioxidant compounds

Methods of chemical synthesis of Mitogenome-C3, Mitogenome-C5 and Mitogenome-C15, reflected in figure 2 and described below. The spectra of nuclear magnetic resonance obtained from the use of the instrument Varian 300 MHz. For1H-NMR internal standard in CdCl3 ,was tetramethylsilane was. For31P NMR external standard was 85%phosphoric acid. Chemical shifts (δ) are presented in mlnd relative to the standard. Elemental analyses conducted Campbell Microanalytical Laboratory, University of Otago. Electrospray mass spectrometry is carried out using liquid chromatography mass spectrometer LCMS-QP800X. Royal solutions were prepared in absolute ethanol and kept at -20°C in the dark.

Mitchison-C3 (6). The scheme of synthesis of Mitogenome-C3 shown in figa, Source material, 2,3,4,5-tetramethoxysilane (1) (Lipshutz, B.H., Kim, S.-k., Mollard, P. And Stvens, K.L. (1998) tetrahedron 54, 1241-1253) obtained by the reduction of 2,3-dimethoxy-5-methyl-1,4-benzoquinone (CoQ) to hydrochinone (Garpino, L.A., Triolo, S.A. and Berglund, R. A. (1989) J. Org. Chem. 4, 3303-3310) c followed by methylation with obtaining 1 (Lipshutz, B.Y., Kim, S.-k., Mollard, P. And Stevens, K.L. (1998) Tetrahedron 54, 1241-1253). Solution 1 (6,35 g, and 29.9 mmol) in dry hexane (80 ml) and N,N,N',N'-tetramethylethylenediamine (8.6 ml) was placed in a cylindrical mixer with a fiery drying in the tube Slinka (Schlenk) c flame dried under a nitrogen atmosphere. A solution of n-utility (1.6 M, 26,2 ml) in hexane is added slowly at room temperature and the mixture is cooled and stirred at 0°C for 1 hour. After cooling to -78°C. add dry tetrahydrofuran (THF) and a small amount of the reaction mixture are removed, the reaction of cut D2Oand investigate method1H NMR to confirm the methylation reaction. Then the yellow suspension is transferred into a second tube Slinka with flaming drying, containing CuCN (0.54 g, 6,03 mmol)under nitrogen atmosphere at -78°C. the Mixture is heated to 00C for 10 minutes, then cooled to -78°C, add allylbromide (3,62 ml), the reaction mixture was stirred overnight (19 hours) and allow to warm it to room temperature. Reaction cut of 10% aqueous NH4Cl (75 ml) and extracted with simple ether (2×200 ml). The combined ether extracts are washed with H2About (2×150 ml), 10%aqueous NH4OH (200 ml) and saturated aqueous NaCl (200 ml). Organic solvents were dried over MgSO4, filtered and the solvent from Aleut rotary evaporation under vacuum, in the result, get the crude product (7,25 g). Column chromatography on silica gel and elution 20%mixture of simple ether/hexane gives pure 1,2,3,4-tetrametoksi-5-methyl-6-(2-propenyl)benzene (2) (Yoshioka, T., Nishi, T., Kanai, T., Aizawa, Y., Wada, K., Fujita, T. And Horikoshi, H., (1993), Eur. Pat. Appl, EP 549366 A1) (6,05 g, 83.5 per cent).1H NMR: δ of 5.84 is 5.98 (1H, m, -CH=C), 4,88-to 5.03 (2H, m, =CH2), 3,78, 3,80, 3,90, 3,92 (12H, s, OMe), to 3.38 (2H, d, J=7,0 Hz, Ar-CH2), and 2.14 (3H, s, Ar-Me) mind

A solution of 2 (8.0 g, 33,05 mmol) in dry THF (45 ml) is added dropwise over 20 minutes under argon to a stirred suspension of 9-borabicyclo[3,3,1]nonane in THF (79 ml, 39,76 mmol, 0.5 M) at 25°C. the Resulting solution is stirred over night at room temperature and for 2 hours at 65°C under argon. The mixture is then cooled to 0°C and poured dropwise 3M NaOH (53 ml), and then 30% aqueous H2About2(53 ml). After 30 minutes stirring at room temperature the aqueous phase is saturated with NaCl and extracted 3 times THF. The combined organic fractions washed with saturated aqueous NaCl, dried (Na2SO4), filtered and evaporated, resulting in getting oily residue (11.5g), which is purified column chromatography on silica gel (200 g, a compacted mixture of simple ether/hexane 1:9). Elution with a mixture of simple ether/hexane 1:4 gives pure 3-(2,3,4 .5-tetrametoksi-6-were)propan-1-ol (3) as a viscous colorless oil (6.5 g, 80%).1H NMR: δ 3,91, 3,90, 3,84, 3,79 (12H, s, OMe), of 3.56 (2H, t, J=7,0 Hz, -CH2-OH), of 2.72 (2H, t, J=7,0 Hz, Ar-CH2), 2,17 (3H, s, Ar-Me), of 1.74 (2H, quintet, J=7,0 Hz, -CH2-) mind Elemental analysis: calculated for C14H23O5C 62,2; H 8,2. Found: 62,2; N 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. Added dropwise methanesulfonanilide (1.8 g , 1.20 ml of 15.75 mmol) in CH2Cl2(50 ml) for 20 minutes and the reaction mixture was stirred at room temperature for 1 hour. The mixture is then diluted with CH2Cl2(50 ml) and the organic layer was washed with H2About (5×100 ml), 10% aqueous solution of NaHCO3(100 ml), dried (MgSO4), filtered and the solvent is removed under vacuum on a rotary evaporator, resulting in a gain of 1-(3-methanesulfonylaminoethyl)-2-methyl-3,4,5,6-tetramethoxybenzene (4) in liquid form (4.8 g, 95%).1H NMR: δ 4,27 (2H, t, J=7,0 Hz, -CH2-O-SO2-Me), 3,91, 3,89, 3,82, 3,78 (12H, s, OMe), 3,03 (3H, s, -O-SO2-Me), 2,70 (2H, t, J=7,0 Hz, Ar-CH2-), 2,17 (3H, s, Ar-Me), 1,9 (2H, m, -CH2-) mind

The crude methanesulfonate 4 (3,30 g, 9.8 mmol) are used directly in the next reaction, mixing with svejeistolcheng mixture of triphenylphosphine (4,08 g, 15.6 mmol) and NaI (7.78 g, with 51.9 mmol) in the tube Kimaks (Kimax) and seal it under argon. The mixture is then kept at the tempo is the atur 70-74°C, stirring with a magnetic stirrer for 3 hours, during which time the mixture changed from molten viscous liquid to a glassy solid. The tube is cooled to room temperature and the residue is stirred with CH2Cl2(30 ml). Then the suspension is filtered and the filtrate evaporated to dryness. The residue is dissolved in a minimum amount of CH2Cl2and triturated with an excess of simple ether (250 ml) until precipitation of a white precipitate. The solid precipitate was filtered and washed with simple ether, dried in vacuum and get pure [3-(2,3,4,5-tetrametoksi-6-were)propyl]triphenylphosphonium (5) (5,69 g, 90%).1H NMR: δ 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-R+), 2,98 (2H, t, J=7,0 Hz, CH2-Ar)to 2.13 (3H, s, Ar-Me), 1,92-of 1.78 (2H, m, -CH2-) ppm,31P NMR (121,4 MHz) δ 25,32 mind Elemental analysis: calculated for C32H36IO5P: C to 59.8; H 5,7; P 4,8; found: C to 59.8; H 5,8; P 4,5%.

The solution of iodide 5 (4,963 g, 7.8 mmol) in CH2Cl2(80 ml) was shaken with 10%aqueous solution of NaNO3(50 ml) in a separating funnel for 5 minutes the Organic layer is separated, dried (Na2SO4), filtered and evaporated under vacuum, resulting in a gain of nitrate salt 5 (4.5 g, 7.8 mmol, 100%), which is dissolved in a mixture of CH3CN and H2O (7:3, 38 ml) and stirred at 0°C in an ice bath. Then add pyridine-2,6-dicarbon the new acid (6.4 g, 39 mmol), and then poured dropwise a solution of cerium ammonium nitrate (21,0 g, 39 mmol) in CH3CN/H2O (1:1, 77 ml) for 5 minutes. The reaction mixture was stirred at 0°C for 20 minutes and then at room temperature for 10 min. Then the reaction mixture was poured into H2O (200 ml) and extracted with CH2Cl2(200 ml), dried (Na2SO4), filtered and evaporated under vacuum, resulting in a gain of crude nitrate [3-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadiene-1-yl)propyl]triphenylphosphine (6). The entire product was dissolved in CH2Cl2(100 ml) and shaken for 10 min with 20% aqueous KBr solution (50 ml). The organic layer is separated, dried and evaporated under vacuum, resulting in a gain bromilow salt 6 (4.1 g, 93,6%),1H NMR δ of 7.90-7,65 (15H, m, Ar-H), 4,15-of 4.05 (2H, m, CH3-R+), 3,96, 3,95, (6H, s, OMe), with 2.93 (2H, t, J=7,0 Hz, CH2-Ar)to 2.15 (3H, s, Ar-Me), 1.85 to to 1.70 (2H, m, -CH2-) mind,31P NMR δ 25,29 mind

A solution of bromide 6 (3,65 g, 6.5 mmol) in CH2Cl2(75 ml) was shaken with 10% m/o aqueous solution of methanesulfonate sodium (100 ml) in a separating funnel for 5 minutes. CH2Cl2layer is separated, dried (Na2SO4), filtered and evaporated under vacuum, resulting in a gain methansulfonate salt [3-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadiene-1-yl)propyl]triphenylphosphine (6 (3.7 g, 98%).1H NMR: δ 7,88-of 7.60 (15H, m, Ar-H), 3,93, 3,92, (6H, s, OMe), 3,90-of 3.78 (2H, m, CH2-P+), 2,85 (2H, t, J=7,0 Hz, CH2-Ar), 2,70 (3H, s, OSO2CH3), is 2.09 (3H, s, Ar-Me), 1,82 by 1.68 (2H, m, -CH2-) mind,31P NMR (121,4 MHz) δ 25,26 mind Elemental analysis. Calculated for C31H33O7PS: C 64,1; H 5,7; P 5,3; S 5,5. Found: C To 63.8; H 5,9; S To 5.3; P 5,2%.

Mitchison-C5 (14). The scheme of synthesis of Mitogenome-C5 shown in figv. Dihydropyran (46,83 g, 0.55 mmol) are added to 2,3-dimethoxy-5-methyl-1,4-benzohydroquinone (CoQ0) (50 g, 0,275 mol)dissolved in acetic acid (500 ml), and stirred at room temperature for 10 minutes. To this solution add BF3.Et2O (38,57 g, 0,271 mol). The resulting solution was stirred at room temperature. After this time the crude reaction mixture is poured into ice water (500 ml) and extracted with chloroform (1000 ml). The organic extract was washed with saturated salt solution (500 ml) and dried (MgSO4). The solvent is removed under vacuum and get soggy 2,3-dimethoxy-5-methyl-6-(tetrahydropyran-2-yl)-4-(tetrahydropyran-2-yloxy)phenol (7) in the form of oil red (115 g), which is used without further purification. A solution of crude 7 (110 g) in a mixture of acetic acid/perchloric acid (a 97.5:2.5 a, 500 ml) hydronaut over 5% palladium/charcoal (5,42 g) at atmospheric pressure and room temperature until the absorption of hydrogen for three days). Then the reaction mixture was filtered through celiby filter and the solid residue washed with ethanol (500 ml). The combined filtrate is divided into three equal parts and each part is added to distilled water (1000 ml) and extracted with CH2Cl2(2×200 ml). The combined organic extracts washed with saturated salt solution (500 ml), saturated sodium bicarbonate solution (500 ml), saturated salt solution (300 ml)and dried (MgSO4). The mixture is then filtered and the solvents removed under vacuum, resulting in a gain of crude 4-acetoxy-3-(5-acetoxyethyl)-5,6-dimethoxy-2-methylphenylacetic (8) in the form of oil red (110 g), which is used in the subsequent step without further purification.1H NMR: δ 4,0-to 4.15 (2H, m, -CH2-O), 3,86 (6H, s, 2x OMe), 2,58 (2H, t, J=7,0 Hz, -CH2Ar), a 2.12 (3H, s, Ar-Me), to 2.06 (6H, s, 2x CH3-C=O)2,02 (3H, s, CH3-C=O), 1,35 is 1.70 (6H, m, -CH2CH2CH2-) mind

Sociallyengaged (8.0 g, 0.21 mol) is added to dry THF (500 ml) in a 1 liter round bottom flask, equipped with magnetic stirrer, reflux condenser and immersed in a water bath at room temperature. A solution of crude 8 (74 g) in dry fresh THF (100 ml) dropwise poured to a mixture of THF/LiAlH4during the period of time 25-30 minutes. Add additional dry THF (200 ml) to facilitate stirring and the reaction to shift the ü leave to mix for 5 hours at room temperature. Then the reaction being removed by adding dropwise 3M HCl (20 ml) followed by slow addition of distilled water (70 ml). Then the reaction mixture is filtered and the filtrate washed with saturated salt solution (2×300 ml), dried (MgSO4), filtered and the solvent is removed under vacuum. The green residue remaining on the filter funnel, dissolved in 15% HCl (500 ml) and CH2Cl2(1×300 ml, 2×200 ml). The organic fractions are combined and washed with saturated salt solution (400 ml), dried (MgSO4), filtered and evaporated under vacuum. This extract is combined with the material from the processing phase of the filtrate and obtain crude 2-(5-hydroxyphenyl)-5,6-dimethoxy-3-methylbenzo-1,4-diol (9) (68,3 g) in the form of oil red. This product 9 purified using column chromatography on silica gel (600 g, compacted with a 10% mixture of simple ether/ CH2Cl2). Elution with 10% mixture of simple ether/ CH2Cl2gives some amount of unreacted 8 and starting material 2,3-dimethoxy-5-methyl-1,4-benzohydroquinone. Elution with 20% mixture of simple ether/ CH2Cl2gives a mixture of 9 and quinone 10 (14,14 g, 19% of 2,3-dimethoxy-5-methyl-1,4-benzoquinol). Connection 9 slowly turns into a quinone 10 when standing on the air, and satisfactory elemental analysis could not be obtained.1H NMR: δ 5,41 (1H, s, Ar-OH), 5,38 (1H, s, Ar-OH), and 4.8 (6H, s, 2 x Ar-OMe), the 3.65 (2H, t, J=6.3 Hz, CH2-OH), 2,61 (2H, t, J=6,4 Hz, Ar-CH2), and 2.14 (3H, s, Ar-Me), 1,42 by 1.68 (6H, m, 3-CH2-) mind

The solution ginola 9 (7.5 g, 27.7 mmol) in CH2Cl2(150 ml) saturated with gaseous oxygen at atmospheric pressure and add a solution of NO2in CH2Cl2(1 ml, 1,32 M). The reaction mixture was stirred at room temperature under the atmosphere of oxygen for 18 hours, by which time TLC (40 % simple ether/CH2Cl2) shows that the formation of quinone 2-(5-hydroxyphenyl)-5,6-dimethoxy-3-methyl-[1,4]benzoquinone (10) is completed. Then the solvent is removed under vacuum and get the product 10 (Yu, C.A. and Yu, L. (1982) Biochemistry 21, 4096-4101) (7,40 g) in the form of oil red.1H NMR: δ 3,99 (6H, s, 2 x Ar-OMe), the 3.65 (2H, t, J=6.3 Hz, CH2-OH), 2,47 (2H, t, J=6.3 Hz, Ar-CH2), a 2.01 (3H, s, Ar-Me), 1,52 is 1.60 (2H, m, -CH2-), 1,37 was 1.43 (4H, m, -CH2CH2-) mind

Prepare a solution of 10 (7,40 g and 27.3 mmol) in CH2Cl2(150 ml) and triethylamine (5,46 g, 5.46 mmol) and add to it a solution of methanesulfonamide (2,48 g, 30 mmol) in CH2Cl2(50 ml) over 30 minutes under stirring. After stirring for an additional 1.5 hours at room temperature the reaction mixture is washed with distilled water (5×100 ml), saturated sodium bicarbonate solution (150 ml) and dried (MgSO4). The mixture is filtered and the solvent is removed under vacuum, in achiev is Tate which receive the crude methanesulfonate (9,03 g) in the form of oil red. 1H NMR: δ 4,19 (2H, t, J=7.5 Hz, -CH2-OMc), of 3.95 (6H, s, 2xAr-OMe), 2,98 (3H, s, OSO2CH3), is 2.44 (2H, t, J=7.5 Hz, Ar-CH2-), to 1.98 (3H, s, Ar-Me), a 1.75 (2H, quintet, J=7.5 Hz, -CH2-), to 1.38 to 1.48 (4H, m, -CH2-CH2-) mind Methanesulfonate was dissolved in 10% (m/o) of NaI in acetone (100 ml) and stirred for 44 hours at room temperature. The mixture is then concentrated under vacuum and to the residue add H2O (100 ml). The mixture is extracted with CH2Cl2(3×70 ml) and the combined organic extracts washed with a saturated solution of salt, dried (MgSO4), filtered and the solvent is removed under vacuum, resulting in a gain of crude 2-(5-iopentol)-5,6-dimethoxy-3-methyl-[1,4]benzoquinone (11). This product is purified column chromatography on silica gel (150 g). Elution CH2Cl2and a 10% mixture of simple ether/ CH2Cl2give pure 11 (7,05 g, 69%) as oil red.1H NMR: δ 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), of 1.85 (2H, quintet, J=7.5 Hz, -CH2-), to 1.38 to 1.48 (4H, m, -CH2-CH2-) mind Elemental analysis. Calculated for C14H19IO4: FROM 44.5; H 5,1; I 33,6. Found: From 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 NaBH4(0.16 g, 4.3 mmol) and the mixture becomes colorless within 1 min. After 5 minutes at room temperature add 5% aqueous HCl solution (100 ml)and the solution extracted with CH 2Cl2(2×50 ml). The organic fractions combined, dried (MgSO4), filtered and the solvent is removed under vacuum, resulting in a gain of 12 (1,15 g, 100%) as sensitive to the effects of oxygen yellow oil which is used without delay.1H NMR: δ are 5.36, 5,31 (2H, s, Ar-OH), to 3.89 (6H, s, 2x Ar-OMe), 3,20 (2H, t, J=7.5 Hz, -CH2-I), 2,62 (2H, t, J=7.5 Hz, -CH2-Ar)to 2.15 (3H, s, Me), 1,82-of 1.92 (2H, m, -CH2-), 1,45-1,55 (4H, m, -CH2-CH2-) mlnd a Mixture of 12 (1,15 g, 2,87 mmol) and triphenylphosphine (1.2 g, or 4.31 mmol) is placed in the tube Kimaks (Kimax) with a cylindrical stirrer. The tube is rinsed with argon, tightly closed, heated and stirred for 14 hours at 700C. Formed a dark solid, which was dissolved in CH2Cl2(10 ml) and triturated with simple ether (200 ml), the resulting white precipitate quickly filtered out. The precipitate, which becomes sticky under the influence of air, re-dissolved in CH2Cl2and evaporated under vacuum, resulting in getting the crude product iodide [5-(2,5-dihydroxy-3,4-dimethoxy-6-were)pentyl]triphenylphosphine (13) (2,07 g, 115%) as a brown oil. The material is not stable when stored for extended periods of time and used as soon as it becomes available for subsequent reactions.1H NMR: δ 7,84-7,68 (15H, m, Ar-H), the 5.45 (1H, s, Ar-OH), to 5.35 (1H, s, Ar-OH), to 3.89 (3H, s, Ar-OMe), 387 (3H, s, Ar-OMe), the 3.65 (2H, m, -CH2-+PPh3), to 2.54 (2H, t, J=7,0 Hz, Ar-CH2), of 2.08 (3H, s, Ar-Me), of 1.65 and 1.75 (2H, m, -CH2-), 1,45-1,55 (4H, m, -CH2CH2-) mind,31P NMR δ 25,43 mind

A solution of 13 (2,07 g) in CH2Cl2(50 ml) saturated with gaseous oxygen and add a solution of NO2in CH2Cl2(0.5 ml, 1,32 M). Then the reaction mixture was stirred at room temperature under oxygen atmosphere for 18 hours. The solvent is removed under vacuum and get the crude product iodide [5-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadiene-1-yl)pentyl]triphenylphosphine (14) in the form of oil red. The residue is re-dissolved in CH2Cl2(10 ml) and triturated with simple ether (200 ml), resulting in getting the original yellow residue, which after a few minutes thickens in the oil red. The solvent is drained and the residue dissolved in CH2Cl2the solvent is removed under vacuum and get the product (14) (1,866 g) in the form of oil red. Aliquot amount (0,880 g) 14 purified column chromatography on silica gel (20 g). Elution CH2Cl2gives a certain amount of unidentified material purple. Elution with a mixture of 5% ethanol/ CH2Cl2gives pure iodide 14 (0,606 g) in the form of oil red.1H NMR: δ 7,84-7,68 (15H, m, Ar-H) 3,98 (6H, s, 2xAr-OMe), the 3.65 (2H, m, CH2-P ), is 2.40 (2H, t, J=7.5 Hz, Ar-CH2), from 2.00 (3H, s, Ar-Me), 1,71 (4H, m, -CH2-), was 1.43 (2H, m, -CH2-) mind,31P NMR (121,4 MHz) δ 25,47 mind Elemental analysis. Calculated for C32H36IO4P: FROM 59.8; H 5,7; I 19,8; R IS 4.8. Found: 60,0; N 5,3; I 19,7; R 4,7%.

Mitchison-C15 (16). The scheme of synthesis of Mitogenome-C15 shown in figs. The solution of K2S2O8(0,450 g of 1.66 mmol) in N2O (25 ml) was poured dropwise over 2.5 hours to a stirred suspension AgNO3(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 N2A:CH3CN (1:1, 36 ml)maintained at a temperature of 75°C. After stirring for 10 minutes the mixture is cooled and extracted with a simple ether (4×30 ml). The combined organic phase was washed with H2About (2×100 ml), NaHCO3(1M, 2×50 ml) and saturated NaCl (2 x 50 ml). The organic phase is dried (Na2SO4), filtered and concentrated under vacuum, resulting in receive oil red (0,444 g). Column chromatography of the crude oil (silica gel, 15 g) and elution with mixtures of CH2Cl2and simple ester(0%, 5%, 20%) gives 2-(15-hydroxypentanal)-5,6-dimethoxy-3-methyl-[1,4]benzoquinone (15) (0,192 g, 33%) as oil red.1H NMR: δ 3,99, 3,98 (6H, s, OMe), to 3.64 (2H, t, J=6.5 Hz, -CH2OH), a 2.45 (2H, t, J=7.5 Hz, -CH2-ring), 1,4-1,2 (26H, m, -(CH2)13-). Element and the Alize. Calculated for C24H40About5: FROM 70.6; H 9,9. Found: 70,5; H 9,8%.

A mixture of triphenylphosphine (of 0.066 g, 0.25 mmol), Ph3PHBr (0,086 g, 0.25 mmol) and 15 (0,101g, 0.25 mmol) is stirred in an argon atmosphere in a sealed tube Kimaks at 70°C for 24 hours, by which time it turns into a viscous red oil. The residue is dissolved in a minimum amount of CH2Cl2(0.5 ml) and poured into a simple ether (10 ml) to give an oily precipitate of red. The solvent is then drained and the residue dissolved in CH3HE (0.5 ml) and diluted with N2About (10 ml)containing 48% HBr (1 drop). A red precipitate is formed after deposition of the sediment top layer is drained and the residue is washed with water (5 ml). Then the residue is dissolved in ethanol (5 ml) and the solvent is removed under vacuum. The residue re-dissolved in CH2Cl2(0.5 ml), diluted with simple ether (5 ml), the solvent is drained and the residue is placed in vacuum (0.1 mbar) for 24 hours, resulting in a gain bromide [15-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadiene-1-yl)pentadecyl]triphenylphosphine (16) (0,111 g, 61%) as a foam product is yellow, which turns into a red oil in contact with air.1H NMR: (299 MHz) δ of 7.6 to 8.0 (15H, m, Ar-H), to 3.89 (6H, s, OMe), 3,9 (2H, m, -CH2-P), and 2.6 (2H, m, -CH2-ring), 1,7-1,1 (26H, m, -(CH2)13-) mind,31P NMR (121,4 MHz) δ 25,7 mind Electrospray mass spectroscopy found (M+) 653 calculated for C42H54About4R+653. Analytical data obtained during the combustion, are unsatisfactory due to unstable levels of solvents.

Example 3. Properties explanatory directed insertion in mitochondria antioxidant compounds

The present invention recognizes that in order to be suitable for a wide range of applications, for example, for compounds of dosage forms such as tablets, there is an advantage due to the ability to form crystalline or solid form directed insertion in mitochondria antioxidant compounds. Similarly, the authors of the invention, without wishing to be bound by any theory, I believe that the antioxidative action of the compounds of the present invention is due, at least partially, their physico-chemical properties.

The distribution coefficients for various antioxidant compounds are presented in table 1. The distribution coefficients Octan-1-ol/CBE determined by adding 400 nmol connection to 2 ml saturated PBS Octan-1-ol and the mixture for 30 minutes at 37°C With 2 ml saturated octane-1-I CBE. Concentration of the compound in the two phases measured by UV absorption at the wavelength of the s 269 nm and quantitatively calculated from the standard curve of compounds in the saturated Oxton-1-I CBE or saturated PBS octane-1-Ola (Kelso, G.F., Porteous, C.M., Couler, C.V., Hughes, G., Porteus, W.K., Ledgerwood, E.C., Smith, R.A.J., and Murphy, M.P., 2001, J. Biol Chem. 276, 4588; Smith, R.A.J., Porteous, C.M., Coulter, C.V. and Murphy, M.P. 1999, Eur. J. Biochem. 263, 709). Royal solutions of the compounds prepared in absolute ethanol and stored at -20°C in the dark. [3H]TPMP obtained from American Radiolabelled Chemicals Inc. (MO, USA).

Particularly noteworthy is the low coefficient of distribution of compounds with a small number of carbon atoms in the bridge connecting the molecule antioxidant and phosphonium. For example, the connection according to the present invention, mentioned in the description as Metochion-C3, which has a bridge of 3 carbon atoms, characterized by the distribution coefficient is approximately 50 times less than that observed for related compounds, Mitogenome-C10 (table 1).

dto $ 7.91×105
Table 1
The distribution coefficients of antioxidants and related compounds
ConnectionThe distribution coefficient
Methyltriphenylphosphonium (TRMR)and0,35±0,02
Mito Vit Eb7,4±1,6
4-Bromotetradecaneb383±0,22
4-Iodocyclopentanec4,0±0,4
Mitchison-C15
Mitchison-C10and160±9
Mitchison-C513,9±1,9
Mitchison-C3c2,8±0,3
α-tocopherolb27,4±1,9
Brazillionaired310±60
Idebenonedof 3.1×103
Dellusionaldof 3.1×105
Coenzyme Q0d1,33
Coenzyme Q1d409
Coenzyme Q2dof 4.44×104
Ubiquinone (Coenzyme Q10)dis 1.82×1020
Originalda 4.53×1020
Dellusional
Idebenoldof 7.82×103

Dataa-Care the distribution coefficients in the system Octan-1-ol/saline phosphate buffer is defined at 25°C or 37°C as described above, or the distribution coefficientsdin the system octanol/water calculated by the program Advanced Chemistry Development (ACD) Software Solaris V4, 67, as described in the publication Jauslin, M.L., Wirth, T., Meier, T., and Schoumacher, F., 2002, Hum. Mol. Genet 11, 3055.

aKelso, G.F., Porteous, C.M., Coulter, C.V., Hughes, G., Porteus, W.K., Ledgerwood, E.C., Smith, R.A.J., and Murphy, M.P., 2001, J. Biol Chem. 276, 4588.

bSmith, R.A.J., Porteous, C.M., Coulter, C.V. and Murphy, M.P. 1999, Eur. J. Biochem. 263, 709

cSmith, R.A.J., Porteous, C.M., Gane, M., and Murphy, M.P. 2003, Proc. Nat. Acad. Sci., 100, 9, 5407.

Of the distribution coefficients of Oxton-1-ol/CBE clear that Metochion-C3, Mitchison-C5, Mitchison-C10 and Mitchison-C15 cover a wide range of hydrophobicity. In this Metochion-C3 is similar to simple, relatively water-soluble cation TRMR, whereas data for Mitogenome-C15 indicate that it has a very low solubility in water. As you know, the cations alkyldiphenylamine, such as Mitogenome, is absorbed on the phospholipid double layers with the cation at the level of carboxylic acid groups, while the hydrophobic alkyl group penetrates into the hydrophobic core of m is mpany. I believe that the longer a methylene bridge, the deeper originally antioxidant will penetrate into the hydrophobic core of the membrane. The authors believe that these compounds would be the maximum degree to which penetration occurs in one leaflet of the membrane, as shown in figure 3, which presents options Mitogenome, United with a typical phospholipid. These models show that the fragment of original of Mitogenome-C3 only penetrates close to the membrane surface layers, whereas for Metochion-C10 and Mitogenome-C15 depth of this penetration is close to the core of the phospholipid double layer.

The inventors have synthesized a number of antioxidant compounds with different hydrophobicity and depth of penetration into phospholipid double layer.

Example 4. Mitochondrial uptake directed input connections

To show that the direction of introduction into the mitochondria is effective, determine uptake by mitochondria as a response to the membrane potential explanatory antioxidant compounds Mitogenome-C3, Mitogenome-C5, Mitogenome-C10 and Mitogenome-C15.

To measure the absorption of antioxidant compounds excited mitochondrion designed ion-selective electrode (Smith, R.F., Kelso, G.F., James, A.M. and Murphy, M.P. (2004) Meth. Enzymol, 382, 45-67; Davey, G.P., Tipton, K.F. and Murphy, M.P. (1992) Biochem. J. 288,439-443; Kamo, N., Muratsugu, M., Hongoh, R. and Kobatake, Y. (1979) J. Membr. Biol. 49, 105-121)/ Electrode and the reference electrode Ag/AgCl put through airtight Perspex cover in 3 ml of the incubation chamber, provided with stirring and temperature control at 30°C, equipped with an injection hole for adding substrates. To measure the absorption of antioxidant compounds mitochondria rat liver (1 mg protein/ml) were incubated at 30°C in the environment KCl (120 mm KCl, 10 mm HEPES, pH of 7.2, 1 mm EGTA), nigericin (1 μg/ml) and rotenone (8 µg/ml). Added succinate (10 mm) and FCCP (500 nm)where indicated. The signal from the ion-selective electrode is held in the data collection system PowerLab Data through front power pH and analyzed using a computer program Chart, all the equipment from ADInstruments.

Mitochondria rat liver prepared by homogenization followed by differential centrifugation in ice buffer containing 250 mm sucrose, 5 mm Tris-HCl, 1 mm EGTA, pH 7,4 (Chappel, J.. and Hansford, R.G. (1972) in his book Subcellular components: preparation and fractionation, pp.77-91 (Birnie, G.D., Ed.) Butterworths, London). Determine the concentration of protein by the biuret assay using BSA as standard (Gornall, A.G., Bardawill, C.J. and David, M.M. (1949) J. Biol. Chem. 177, 751-766). Mitochondrial membrane potential was measured by adding 500 nm TRMR, supplemented with 50 nCi of[3H]TPMP, mitochondria suspended in the medium KCl (120 mm KCl, 10 mm HEPES, pH of 7.2, 1 mm EGTA) at 25°is (Brand, M.D. (1995) in the book: Bioenergetics - a practical approach, pp.39-62 (Brown, G.C. and Cooper, C.E., Eds.) IRL, Oxford). After incubation of mitochondria tabletirujut by centrifugation, and determine the number of [3H]TPMP in the upper layer and tablets scintillation counter, and calculate the membrane potential, taking mitochondrial volume of 0.5 µl/mg of mitochondrial protein and the amendment to TRMR binding of 0.4 (Brown, G.C. and Brand, M.D. (1985) Biochem. J. 225, 399-405).

The inventors have designed ion-selective electrodes for the measurement of their concentrations in the steady state (Smith, R.A., Kelso, G.F., James, A.M. and Murphy, M.P. (2004) Meth. Enzymol. 382, 45-67; Davey, G.P., Tipton, K.F. and Murphy, M.P. (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 on simple cations triphenylphosphine, such as TRMR is Nernstian with a linear dependence of the voltage on the electrode from log10[concentration of cation] and slope of ~ 60 mV at 30°C (Davey, G.P., Tipton, K.F. and Murphy, M.P. (1992) Biochem. J. 288, 439-443; Kamo, N., Muratsugu, M., Hongoh, R. and Kobatake, Y. (1979) J. Membr. Biol. 49, 105-121). The most hydrophilic compound, Mitchison-C3, also gives Nernstian dependence for the electrode with a slope close to 60 mV at concentrations above 10 mm. This is shown in figa, right, in the form of a logarithmic voltage on the electrode of the serial additions 1 PM Mitogenome-C3 in the absence of mitochondria. For Mitogenome-C5, Mituhiro the a-C10 and Mitogenome-C15 electrode also reacted quickly and stably on successive addition in the absence of mitochondria (FIGU, 4C and 4D, respectively, the chart to the right). However, in these cases, the response of the electrode were not Ninstance, as the authors suggest, due to the greater hydrophobicity of these compounds. Even if so, all four of antioxidant compounds ion-selective electrode allows to measure the free concentration of compounds and, thus, their uptake by mitochondria in real time.

To measure the absorption of antioxidant compounds mitochondria injected into the electrode chamber in the presence of rotenone to prevent membrane potential (left part of figure 4). Then make five consecutive additions of antioxidant compounds at 1 μm to calibrate the response of the electrode and succinate respiratory substrate to generate a membrane potential. Mitochondrial stimulation leads to rapid absorption of all types of antioxidant compounds mitochondria, and subsequent addition of decoupling FCCP destroys the membrane potential and leads to its rapid release from the mitochondria (figa-I, left). These experiments show clearly dependent voltage absorption mitochondrial membrane Mitogenome-C3, Mitogenome-C5 and Mitogenome-C10. And if Mitchison-C15 also absorbed by the mitochondria at the stage of induction of membrane potential, the response of the electrode n is Mitchison-15 in the presence of mitochondria was weaker, more noisy more prone to shift. This is different from the electrode response for Mitogenome-15 in the absence of mitochondria (see chart right) and due to its low free concentrations in the presence of mitochondria.

Then determine the degree of binding antioxidant compounds with unexcited mitochondrion (figure 4, right). For these experiments, different antioxidant compounds were first introduced in the electrode chamber and then injected mitochondria in the presence of rotenone to prevent membrane potential. The decrease in the concentration of antioxidant compounds with the introduction of mitochondria due to the binding of antioxidant compounds with unexcited by the mitochondria. Subsequent addition of succinate to generate a membrane potential indicates dependent membrane potential absorption of compounds, which then becomes reversed when adding FCCP for the destruction of the membrane potential.

The free concentration of Mitogenome-C3 does not change when adding the mitochondria, which indicates that trace amounts of Mitogenome-C3 contact unexcited mitochondrion (figa, right). FCCP-sensitive absorption mitogenome-C3 upon excitation by succinate amounted to approximately 3.7 nmol of mitogenome-C3/mg protein, which matched what was the accumulation rate of 2×10 3. This is consistent with the expected result calculated by the Nernst equation, and mitochondrial membrane potential of about 180 mV, allowing you to introduce amendments on intramitochondrial binding.

For Mitogenome-C5 there is a binding connection with unexcited mitochondria (~0.6 nmol/mg protein), but this is negligible compared with its subsequent absorption after excitation succinate, about 2.8 nmol of Mitogenome-C5/mg protein, corresponding to the accumulation rate of approximately 1.4×103(FIGU, right).

For Mitogenome-C10 significant binding unexcited mitochondria approximately 2.6 nmol of Mitogenome-C10, and this is followed by additional linking approximately 1 nmol/mg of protein when adding succinate (figs, right).

Almost all free Mitchison-C15 associated with unexcited mitochondria, but there is some additional absorption at the excitation succinate. Dependent on membrane potential absorption of Mitogenome-C15 evident in the left diagram fig.4D, where the electrode response is extremely sensitive to be able to measure a small amount of free Mitogenome-C15, when the electrode is calibrated in the presence of mitochondria. In contrast, the absorption of Mitogenome-C15 hard to see on the right side fig.4D where the electrode response is much less sensitive, to allow to measure Mitchison-15 in the absence of mitochondria.

These experiments show that the length of the methylene bridges of antioxidant compounds, at least partially determines their degree of adsorption on the mitochondrial membranes (on the right in figure 4). Adsorption varies from negligible to Mitogenome-C3 until almost complete binding for Mitogenome-C15. When adding Mitogenome-C15 to unexcited mitochondria almost all connection is associated, is distributed on both surfaces of the inner and outer membranes. When induced membrane potential, the authors believe that will be a significant redistribution of the connection drawn to the matrix surface of the inner membrane with the outer surface of the inner membrane and outer membrane. Summarizing, we can say that all types of antioxidant compounds are absorbed by the mitochondria under the action of membrane potential, and the longer methylene bridge, the more their adsorption on the phospholipid double layer.

Example 5. The antioxidant efficiency of explanatory connections, directed insertion in mitochondria

Compounds of the present invention is also very effective against oxidative stress. To measure the antioxidant effectiveness, assess the ability of antioxidant compounds pre order to prevent lipid peroxidation in mitochondria, measured by the accumulation of TBARS in mitochondria under the action of ferrous iron and hydrogen peroxide (figure 5).

In order to quantify the lipid peroxidation, using the TBARS assay. Mitochondria rat liver (2 mg protein/mg) incubated in 0.8 ml of medium containing 100 mm KCl, 10 mm Tris-HCl, pH of 7.6 at 37°C, with the addition of either 10 mm succinate and 8 mg/ml rotenone or ATP regenerating system of 2.5 mm ATP, 1 mm phosphoenolpyruvate and 4 Units./ml pyruvate kinase. Then mitochondria subjected to oxidative stress by adding 50 mm FeCl2/300 mm H2About2for 15 minutes at 37°C. After incubation add 64 ml of 2% (m/o) butylacetamide of hydroxytoluene in ethanol, and then 200 ml of 35% (o/o) HClO4and 200 ml of 1% (m/o) thiobarbiturate acid. Then incubate the samples for 15 minutes at 100°C, centrifuged (5 min at 12000 × g) and the upper layer is transferred into a glass test tube. After adding 3 ml of water and 3 ml of butane-1-ol samples centrifuged and allow to separate the two phases. Then analyze 200 ml sample of the organic layer in fluorometrically anode readout device (λEx= 515 nm; λEm= 553 nm) for the reaction fragments thiobarbituric acid (TBARS) and compared with a standard curve of malondialdehyde (MDA), derived from 0.01 to 5 mm 1,1,3,3-tetraethoxypropane (Kelso, G.F., Porteous, C.M., Coulter, C.V., Hughes, G., Porteous, W.K., Ledgerwood, E.C., Smith, R.A.J. andMutphy, M.P. (2001) J. Biol. Chem. 276, 4588-4596).

For mitochondria excited succinate, the background level of TBARS insignificant, but it increases to about 3.75 mmol MDA/mg protein under the influence of oxidative stress (figa; solid bars). High concentration (5 μm) of any of the antioxidant compounds in substantially prevents the accumulation of TBARS, whereas a simple cation TRMR no. This confirms the fact that originalna side chain mitogenomic antioxidant compounds responsible for antioxidant activity, and no non-specific interaction of the cation with the mitochondria.

In these experiments, succinate will maintain the membrane potential to come true absorption cation mitochondria and also to ensure the transition obikenobi form mitogenomic antioxidant compounds to the active antioxidant originalnoy form (Kelso, G.F., Porteous, C.M., Coulter, C.V., Hughes, G., Porteous, W.K., Ledgerwood, E.C., Smith, R.A.J. and Mutphy, M.P. (2001) J. Biol. Chem. 276, 4588-4596). To see whether the restoration of the respiratory chain antioxidant activity mitogenomic antioxidant compounds, the inventors incubated mitochondria in the presence of ATP and an ATP regenerating system. The hydrolysis of ATP and treatment of mitochondrial ATP synthase leads to extensive pumping of protons, which increases the meme is an early potential in the same way generated by succinate (pigv). This will lead to the same absorption makinosaka antioxidant compounds, as for mitochondria excited succinate, but in this case Methenolone antioxidant compounds will no longer turn in their active originalnye form the respiratory chain. Methenolone antioxidant compounds are ineffective to prevent the peroxidation of lipids in mitochondria, excited by the hydrolysis of ATP (figa, white bars) compared with a significant protection observed in mitochondria induced by succinate (fig.5b, black bars). Therefore, the recovery Mitogenomic antioxidant compounds respiratory chain and accumulation under the action of mitochondrial membrane potential necessary for the manifestation of antioxidant activity mitogenome antioxidant compounds.

Lower levels of lipid peroxidation observed in the control samples of mitochondria induced by succinate, compared with mitochondria, excited ATP (figa). This is due to the protective antioxidant effect of endogenous mitochondrial Fund Coenzyme Q, which remains recovered in the presence of succinate, but is oxidized in the presence of ATP (James, A.M., Smith, R.A. and Murphy, M.P. (2004) Arch. Biochem. Biophys. 423, 47-56; Ernster, ., Forsmark, P. and Nordenbrand, K. (1992) Biofactors 3, 241-8). In summary, we can say that all Methenolone antioxidant compounds require activation of the respiratory chain, in order to be effective antioxidants.

For figa used a concentration of 5 PM for all mitogenomic antioxidant compounds. In order to compare their relative antioxidant efficiency, the inventors was titrated compounds in the presence of succinate: a typical titration shown in figs. The experiment suggests that the antioxidant efficiency of these compounds correlates with the length of the methylene bridge. To quantify this, the authors calculated IC50 values for the prevention of peroxidation of lipids four explanatory mitogenome antioxidant compounds (fig.4D). These measurements confirm that the order of antioxidant effectiveness is: Makinon-C15 > Mitchison-C10 > Mitchison-C5 > Mitchison-C3.

All Methenolone antioxidant compounds accumulate in mitochondria driven by the mitochondrial membrane potential. For the most hydrophobic compounds, Mitogenome-C15, this effect is largely masked by extensive binding to phospholipid double layer. All compounds are effective antioxidants and stable antioxi is based activity more than 15 minutes, require the action of respiratory chain to return antioxidant compound in its active antioxidant form after detoxification intermediates of lipid peroxidation.

Example 6. The influence of the directional input in mitochondria antioxidant compounds on cardiac hemodynamics and mitochondrial function

The impact of the introduction of targeted insertion in mitochondria antioxidant compounds, in particular, Mitogenome-C10 and Mitogenome-C3, on cardiac function assessed using perfusion model Langendorf the isolated heart. Rats are divided in the following four groups introduction: the control (placebo), TRMR (methyltriphenylphosphonium), Mitchison-C10 and Mitchison-C3. After a period of treatment of rats humanely killed, and the extracted heart connect with isolated perfusion system Langendorf. In this system, used retro-perfusion through the aorta to maintain the heart in measuring cardiac activity. The pressure in the left ventricle measured by the balloon in the left ventricle. Also measure coronary flow.

Figure 6 depicts the coronary flow when the pressure in the left ventricle 10 mm Hg for each group of treatment. Coronary flow was measured in predicamental condition and again at time zero minutes, 20 minutes, 40 minutes and 60 minutes, with subsequent induction of ischemia. Spend unidirectional ANOVA and subsequent test bonferroni. Significance depending on predicamento control: *P<0,05; **P<0,01; ***P<0,001. The dependence of importance from the match is a test of time: †P< 0,05; ††P<0,01; †††P<0,001.

The results show that treatment with makinana-C10 significantly restores induced ischemia reduction in coronary flow. Mitchison-C3 has a lesser but still significant effect in the latter time points. The absence of any effect in the introduction TRMR indicates that this molecule antioxidant Mitogenome-C10 and Mitogenome-C3, and not the cation triphenylphosphine responsible for the effects observed for targeted insertion of antioxidant compounds.

7 shows the effect of treatment on diastolic pressure in the left ventricle at 10 mm Hg. Diastolic pressure in the left ventricle was measured before induction of ischemia and again at the zero moment of time, 20 minutes, 40 minutes and 60 minutes with subsequent induction of ischemia. Statistical analysis was ANOVA on ranks with subsequent Dunn test. The dependence of significance from ischemic control: *P<0,05.

† represents P<0,05 through 60 minutes after ischemic control. The results show that treatment with Makinana-C10 leads to a statistically significant increase in diastolic pressure in the left ventricle compared with rats that have not undergone treatment, recovery induced by ischemia reduction in diastolic pressure in the left ventricle. The absence of any effect in the introduction TRMR shows is that what is the molecule of antioxidant Methanone-C10, and not the cation triphenylphosphine, causes the effects observed in the case of directionally injected into the mitochondria antioxidant compounds.

Then determined the impact of the introduction of Mitogenome-C10 and Mitogenome-C3 cardiac rhythm. On Fig presents heart rate for each of the treatment groups before ischemia, at the zero point, 20 minutes, 40 minutes and 60 minutes after induction of ischemia. The presented results are a one-way ANOVA with subsequent test bonferroni. *** represents P<0.001 in comparison with predications control sample. †† represents P<0,05 in comparison with the corresponding postischemic sample. The results show that treatment with Makinana-C10 leads to a significant recovery of the induced ischemia reduction of cardiac rhythm compared with the control rats. The absence of any effect in the introduction TRMR indicates that the molecule antioxidant Mitogenome-C10, and not the cation triphenylphosphine responsible for the effects observed for targeted insertion in mitochondria antioxidant compounds.

Cardiac function was assessed additionally, determining the effect of the introduction of targeted insertion in mitochondria antioxidant compounds on the frequency of contraction and relaxation of the heart. On fightn frequency of contractions in each of the four treatment groups before ischemia, the zero moment of time, 20 minutes, 40 minutes and 60 minutes with subsequent induction of ischemia. On FIGU presents the frequency of relaxation in each of the four treatment groups before ischemia, at the zero moment of time, 20 minutes, 40 minutes and 60 minutes with subsequent induction of ischemia. In each case, conducted the statistical analysis ANOVA on ranks, followed by the test on Dunn. * represents significance with a P<0.05, depending on the respective post-ischemic time for the control samples. †† represents significance with a P<0.01 depending on the respective post-ischemic time for the control samples.

The results show that the introduction of Mitogenome-C10 has a statistically significant effect, restoring induced ischemia decrease in the frequency of contractions and relaxation of the left ventricle compared with the control rats.

The above data clearly show the positive effect from the introduction of targeted insertion in mitochondria antioxidant compounds on cardiac activity. In order to determine the cause of the observed effects on cardiac activity influence directed insertion in mitochondria antioxidant compounds on the function of mitochondria was assessed by the mitochondrial activity in the pre - and post-ischemic condition for each group of treatment. On figa is zobrazen respiratory function of mitochondria, associated with NAD+in the pre - and post-ischemic condition for each of the four treatment groups. On FIGU presents respiratory function associated with FAD in the pre - and post-ischemic condition for each of the four treatment groups. *** represents significance with a P<0,001 depending on the pre-ischemic state of the control rats. ††† is the significance with P<0,001 depending on post-ischemic condition of the control rats.

These data show that Metochion-C10 has a statistically significant positive effect on mitochondrial respiratory function after ischemia compared with control rats. These results confirm the conclusion that the effects from the introduction of targeted insertion in mitochondria antioxidant compounds on cardiac activity due to a protective effect on the function of mitochondria.

Example 7. The stability of complexes of Mitogenome-C10 with β-cyclodextrin.

Preliminary study of Mitogenome-C10 in the form of its bromide established that he will destroy in time in the solid state when stored at 25°C, 50% relative humidity and 40°C, 75% relative humidity. The purpose of this study was to clarify the possibility of increasing stability in the solid state of Mitogenome-C10 by complexation with β-cyclodextrin.

Party M is thenone-C10 No. 6 and idebenone supplied Industrial Research Limited (New Zealand). β-cyclodextrin (lot no R) was purchased from ISP technologies Inc. NaCl, NaH PU and methanol (HPLC) were purchased from BDH.

The study of stability in the solid state net Mitogenome-C10

Sample Mitogenome-C10 (approximately 5 mg) were accurately weighed into clean vials and condition at 25°C, 50% relative humidity, 40°C, 75% RH and 4°C above the silicon dioxide. Bubbles pull through 1, 2, 4, 8, 16, 32 and 64 days and examined for the content of Mitogenome-C10 graded method TSRH using Mitogenome-C10 stored at -20°C over silica gel as a control.

Obtaining complexes Mitchison-C10:β-cyclodextrin

These complexes with different molar ratio (bromide Mitogenome-C10:β-cyclodextrin 1:1, 1:2 and 1:4) was prepared using Mitogenome-C10, party No. 6.

Obtaining a solution of β-cyclodextrin in water

β-cyclodextrin (1,1397 g, corresponding 1,0361 g after adjustment for moisture content) accurately weighed and dissolved in double-distilled water, dispersive ultrasound for 10 minutes, the Volume was adjusted to 100 ml with water.

Obtaining complex Mitchison-C10: β-cyclodextrin (molar ratio 1:1)

Ethanol bromide solution Mitogenome-C10 (90 mg, corresponds 59,95 mg Mitogenome-C10) is evaporated under nitrogen atmosphere on a hot tile supported at 40-50°C, t is the increase in 8 minutes In a glass add a solution of β-cyclodextrin (10 ml) and double-distilled water (30 ml), then dispersed by ultrasound for 40 minutes

Obtaining complex Mitchison-C10: β-cyclodextrin (molar ratio 1:2)

Ethanol bromide solution Mitogenome-C10 (89,8 mg, corresponds to 59.82 mg Mitogenome-C10) is evaporated under nitrogen atmosphere on a hot tile supported at 37-45°C. for 10 minutes, and then for 3 min at 50°C. In a glass add a solution of β-cyclodextrin (20 ml) and double-distilled water (20 ml) and content is dispersed by ultrasound for 30 minutes

Obtaining complex Mitchison-C10: β-cyclodextrin (molar ratio 1:4)

Ethanol bromide solution Mitogenome-C10 (90 mg, corresponds 59,95 mg Mitogenome C-10) is evaporated under nitrogen atmosphere on a hot tile supported 37-50°C for 12 minutes In the glass add a solution of β-cyclodextrin (40 ml)and content is dispersed by ultrasound for 20 minutes

All of the above solutions is frozen and stored at -18°C during the night. Frozen solutions defrost for 2 days using a freeze-dryer LABCONO. Lyophilized solutions stored at -20°C.

Differential scanning calorimetry lyophilized complexes Mitchison-C10:β-cyclodextrin

Differencialnoe scanning calorimetry (DSC) of the three sub is imenovannykh complexes carried out using a differential scanning calorimeter PYRIS-1 firm Perkin Elmer. Sample Mitogenome-C10 prepared by evaporation of an ethanol solution under an atmosphere of nitrogen gas at 35-50°C for 10 minutes

Use aluminum tape (No. 0219-0041 supplied by the company Perkin Elmer). The analysis is performed in a stream of nitrogen. The empty cartridge is used for the baseline.

The temperature interval scan is 50-160°C at the initial temperature to be maintained at 50°C for 1 min, increasing to 160°C at a rate of 10°C /min

Assessment method TSRH

Method TSGH for Mitogenome-C10 developed based on the use of methanol and 0,01M solution of sodium dihydrophosphate (85:15) as mobile phase at a flow rate of 1 ml/min and using UV-VIS detection at 265 nm. As an internal standard using idebenone. Column is the column type Prodigy ODS3 100A (Phenomenex) with a particle size of 5 μm. Later this method was modified after receiving a new column. Mobile phase used in the modified method was methanol and 0,01M solution of sodium dihydrophosphate (80:20). This method was approved. Before analysis of the complexes Mitchison-C10:β-cyclodextrin was tested the influence of β-cyclodextrin on the method TSRH. It is shown that β-cyclodextrin does not affect the assessment of Mitogenome-C10 method TSRH.

The study of stability of complexes Mitchison-C10: β-cyclodextrin

As what was received three sets of Mitogenome-C10 with β-cyclodextrin, the number of Mitogenome-C10 samples weighing 5 mg in different complexes was different. In order to test an equal number of Mithochina-C10 all three complexes were taken of different mass complexes: 4 mg of complex 1:1 with the contents of Mitogenome-C10 1,473 mg, 6.5 mg of complex 1:2 with the contents of Mitogenome-C10 1,469 mg, 11.5 mg of complex 1:4 with the contents of Mitogenome-C10 1,467 mg, and used for stability studies according to the standard procedure (Standard Operating Process.

Volumes of water for TSGH (1.5 ml) was added into the flask with each sample for the complete dissolution of the complexes Mitchison-C10: β-cyclodextrin. Volumes (50 µl) of these solutions were brought to 1 ml with water. Volumes (100 µl) of these diluted solutions of complexes Methine-C10: β-cyclodextrin was mixed with 200 ml of the internal standard solution in methanol concentration of 10 μg/ml Samples were centrifuged for 10 minutes at a speed of 1000 rpm and inject 50 ál of the upper layers in the system TSRH. A standard curve was obtained using solutions of Mitogenome-C10 in the concentration range from 2.5 to 120 mcg/ml, containing 5 mg/ml solution of β-cyclodextrin.

All connections were slightly yellow-orange in colour and very fluffy appearance. Color was not uniform and was more intense in the direction from the bottom flasks for sublimirovanny.

The results of DSC is presented in the following is the form:

Mitchison-C10: In the analysis of a sample of pure Mitogenome-C10 peaks observed at temperatures above 120°C. For one sample of Mitogenome-C10 observed two prominent peak between 130°C and 140°C. In the analysis of another sample, no significant peaks were not observed, but small peaks observed at temperatures above 120°C. After analysis, the vessels were cut and the samples examined. The samples had color from dark green to black in both cases.

β-cyclodextrin: Observed a broad peak in the range between 70°C and 85°C.

Complex Mitchison-C10: β-cyclodextrin (1:1): No significant peaks were observed. After analysis of the cell opened and the sample examined. The color of the sample has undergone a small change to light brown (slight change).

Complex Mitchison-C10: β-cyclodextrin (1:2): No significant peaks were observed. After analysis of the color change of the sample is not observed.

Complex Mitchison-C10: β-cyclodextrin (1:4): No noticeable peaks were not observed, but a very small exothermic peak is observed at 120°C. After analysis, no color change of the sample is not observed.

The appearance of peaks in the sample of pure Mitogenome-C10 indicates that the connection changes when the temperature changes. However, since the peaks were many, and the sample changed color, it is m the Glo to occur due to degradation. In the analysis of the second sample Mitogenome-C10 he gave a thermogram, which differs from thermograms of the first sample. In the case of the complexes was not noticeable peaks or any color change.

The results of the study of stability in the solid state net Mitogenome-C10 (batch No. 3) are given in table 2 and figure 11.

18,37
Table 2
Stability in the solid state of Mitogenome-C10 (batch No. 3)
Transparent glass flaskDayDayDayDayDayDayDay
40°C, 75% relative humidity98,90101,9102,894,0783,2276,7067,25
25°C, 50% relative humidity95,1197,4695,0697,52102,840,76
5°C, the silica gel97,04102,892,9795,6798,3767,3663,70

Stability in the solid state of Mitogenome-C10 (batch No. 3) in the absence of light at 40°C, 75% RH; 25°C, 50% RH and 5°C over blue silica gel. Data represent the average of two values, expressed as a percentage of the initial content.

Due to the significant instability at 25°C, 50% relative humidity compared with the stability at 40°C, 75% relative humidity stability studies were repeated at 25°C, 50% relative humidity with Mitchenerom-C10, party No. 4. The second stability study was conducted in a transparent and darkened flasks and the results presented in table 3 and Fig.

Table 3
Stability in the solid state of Mitogenome-C10 (lot No. 4)
Time (days)124816 3264
Transparent glass flask88,2193,1992,6593,1094,4762,0557,94
Darkened brown glass flask94,8494,52100,2897,6598,0361,4858,66

Stability in the solid state of Mitogenome-C10 (lot No. 4) was measured in the absence of light at 25°C, 50% relative humidity. The results represent the average of three values, expressed as a percentage of the initial content.

Both parties (the parties No. 3 and 4) Mitchison-C10 posed Chemistry Department, showed a sudden drop of content over the 16 days. However, for party No. 4 destruction was not as high in the period from 32 to 64 days compared with the party No. 3. Also observed that the stability of Mitogenome-C10 does not depend on whether the bulb is transparent or darkened.

Mitchison-C10 delivered IRL, was used to obtain complexes Mitchison-C10: β-cyclodextrin. M is thenon-C10, put IRL, was a reddish-yellow syrup in ethyl alcohol. The stability of complexes Mitchison-C10: β-cyclodextrin are shown in table 4 and Fig, 14 and 15. Due to the small number of complexes Mitchison-C10; β-cyclodextrin, available for study, no results found for 1-day and 4-day.

Table 4
Stability in the solid state complexes Mitchison-C10: β-cyclodextrin
Complex 1:1
Time (days)28163264
4°C, silicon dioxide106,38110,97101,71101,71102,68
25°C, 50% relative humidity95,6593,00101,15101,15 108,89
40°C, 75% relative humidity129,22108,77113,48113,4989,25
Complex 1:2
Time (days)28163264
4°C, silicon dioxide105,48101,23105,08111,21101,16
25°C, 50% relative humidity108,1695,46105,41108,55efficiency of 99.78
40°C, 75% relative humidity115,99110,22114,03101,5099,44
Complex 1:4
Time (days)28163264
4°C, silicon dioxide105,10115,86to 100.25107,63107,63
25°C, 50% relative humidity111,46116,0396,6192,4092,40
40°C, 75% relative humidity108,85100,0187,3471,1371,13

Stability in the solid state complexes Mitchison-C10:β-cyclodextrin in the absence of light at 40°C, 75% RH; 25°C, 50% RH and 5°C over blue silica gel. The results represent average values of two quantities, expressed in percent.

The results show that Metochion-C10 can efficiently form complexes with β-cyclodextrin and can be stabilized by complexation with β-cyclodextrin. The results show that Metochion-C10 in complexes with β-cyclodextrin 1:1 and 1:2 was stable in various conditions. The results also indicate that the stability of Mitogenome-C10 in the complex of 1:4 was lower than the stability of Mitogenome-C10 in complexes with β-cyclodextrin 1:1 and 1:2.

Example 8. The stability studies of nelfinavir Mitogenome-C10

The stability of the solution of nelfinavir Mitogenome-C10

The stability of the solution of nelfinavir Mitogenome-C10 was determined in five solvents; water; 0,01M HCl, 0,01M NaOH, IPB (pH of 7.4) and 50% Meon at two temperatures 25°C and 40°C, under two atmospheric conditions, air and nitrogen, for 125 days according to the standard method of study of the applicant.

Prepared solutions of nelfinavir Mitogenome-C10 (100 mg/ml) in five solvents and diluting the stock solution 1 mg/ml nelfinavir of Mitogenome-C10 in the water. Solutions (5 ml) were placed in glass vials, purged with air or nitrogen, closed and placed in storage. Selected aliquot amount (0.25 ml) at 0, 1, 2, 4, 8, 16, 32, 64 and 125 day and method TSRH determined the concentration of Mitogenome-C10.

The results are presented in table 5. The stability of nelfinavir Mitogenome-C10 in 0,01M NaOH is not included in the data, as mesilate of Mitogenome-C10 decomposes in the solvent for 15 minutes. The results show that (a) the stability of the solution does not depend on the atmosphere above rest the rum and (b) temperature has little effect on the stability of Mitogenome-C10 in all solvents, with the exception of HCl.

Table 5
Stability of solutions of nelfinavir Mitogenome-C10 in 4 different solvents under different conditions
ConditionsTime (days)
1248163264125
Water, air, 25°C99,698,398,191,293,898,094,791,7
Water, air, 40°C98,195,094,6to 92.1to 92.191,558,321,8
0,01M HCl, air, 25°C103,7107,6 102,898,398,898,698,692,5
0,01M HCl, air, 40°C98,498,899,296,5100,6104,994,082,6
IPB, air, 25°C95,695,898,694,593,790,489,6100,3
IPB, air, 40°C95,795,594,192,391,2to 89.568,740,0
50% MeOH, air, 25°C97,7to 97.1106,9103,6104,598,6102,850% MeOH, air, 40°C99,298,899,498,5100,5of 101.581,9of 60.5
Water, N2, 25°C106,7USD 114.996,896,799,797,6100,8of 92.7
Water, N2, 40°C97,097,598,393,090,4of 87.359,822,0
0,01M HCl, N2, 25°C102,7112,0103,698, 101,7the 98.999,6br93.1
0,01M HCl, N2, 40°C99,399,6100,298,3to 100.4102,994,1and 88.8
IPB, N2, 25°C99,696,596,9for 95.396,691,990,997,5
IPB, N2, 40°C92,890,9of 92.791,293,3at 88.168,0of 40.3
50% MeOH, N2, 25°C101,297,3105,1104,2102,0105,0101,0100,2
50% MeOH, N2, 40°C99,8100,599,3the 98.9of 101.5103,6is 83.863,6

Data are the average of two values, expressed in percentage of the value at the zero moment of time.

The stability of the solution of nelfinavir Mitogenome-C10 four solvents also presented on Fig, 17, 18 and 19.

Stability in the solid state nelfinavir of Mitogenome-C10

Stability in the solid state nelfinavir of Mitogenome-C10 studied in the absence of light in three different conditions: 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel according to standard methods of tests of the applicant.

A known mass of nelfinavir Mitogenome-C10 placed in transparent glass bottles and stored under different conditions. Two sample taken on 1, 2, 4, 8, 16, 32, 64 and 125 day and determine the concentration of nelfinavir Mitogenome-C10 method TSRH after dissolution of the samples in water. The results are presented in table 6 and Fig.

Mesilate of Mitogenome-C10 stable decomposition of <10%) at 4°C over blue silica gel within 125 days at 25°C./50% relative humidity for 60 days.

br93.1
Table 6
Stability in the solid state nelfinavir of Mitogenome-C10 at 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel
Time (days)1248163264125
4°C over silica gel101,6103,4102,4to 108.2113,596,6of 98.296,0
25°C/50% relative humidity109,2to 110.7110,2108,1107,695,to 91.673,7
40°C/75% relative humidity98,0101,7101,398,586,182,159,9

Data represent the average of two values, expressed in percentage of the value at the zero moment of time.

Example 9. The study of the stability of the complex mesilate of Mitogenome-C10: β-cyclodextrin (1:2)

The stability of the solution of the complex mesilate of Mitogenome-C10 : β-cyclodextrin (1:2)

The stability of the solution of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) define five solvents: water, 0,01M HCl, 0,01M NaOH, IPB (pH of 7.4) and 50% MeOH at two temperatures 25°C and 40°C, in two atmospheric conditions, air and nitrogen for 64 days according to the standard methods of tests of the applicant.

Prepare solutions of complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) (100 mg/ml per mesilate of Mitogenome-C10) in five solvents and diluting the stock solution of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) (1 mg/ml per mesilate of Mitogenome-C10) in the water. Solutions (5 ml) was placed in glass vials, rinsed with air or nitrogen, cover it and put in storage. Selected aliquot amount (0.25 ml) at 0, 1, 2, 4, 8, 16, 32, 64 and 125 day and method TSRH determine concentration.

The results are presented in table 7 and Fig, 22, 23 and 24. The stability of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) in 0,01M NaH not included in the data since the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) decomposes in the solvent for 15 minutes. The results show that (a) the stability of the solution does not depend on the atmosphere above the solution and (b) temperature significantly affects the stability of nelfinavir Mitogenome-C10 in the complex is 1:2 with β-cyclodextrin in all solvents, except for HCl.

Table 7
The stability of the solution of the complex mesilate of Mitogenome-C10-β-cyclodextrin (1:2) in 4 different solvents under different conditions
ConditionsTime (days)
1248163264125
Water, air, 25°C106,5103,396,798,197,997,5100,0105,8
Water, air, 40°C 102,098,7101,3101,492,880,555,917,2
0,01M HCl, air, 25°C96,4100,599,998,6of 101.597,6104,5101,6
0,01M HCl, air, 40°C96,698,4107,1104,0to 100.496,4101,096,2
IPB, air, 25°C97,494,398,396,1107,597,397,399,5
IPB, air, 40°Cfor 95.293,2of 97.898,891,7 87,574,255,9
50% MeOH, air, 25°Cof 97.896,295,897,5102,898,399,7104,5
50% MeOH, air, 40°C99,598,1106,5106,5102,992,684,664,6
Water, N2, 25°C101,198,3102,4103,5109,396,8100,3for 91.3
Water, N2, 40°C100,6100,3/td> 104,8101,694,178,152,814,9
0,01M HCl, N2, 25°C100,1100,797,9101,0103,0100,6104,3100,0
0,01M HCl, N2, 40°C98,696,8103,7104,2100,6of 97.8100,395,14
IPB, N2, 25°C102,097,999,695,6104,693,996,998,7
IPB, N2, 40°Cto 92.193,7for 95.2br93.190,386,6of 54.8
50% MeOH, N2, 25°C105,096,094,196,1106,497,0100,0105,7
50% MeOH, N2x, 40°C98,3the 98.9104,2to 105.399,494,9and 88.864,4

Data represent the average of two values, expressed in percentage of the value at the zero moment of time.

Stability in the solid state complex mesilate of Mitogenome-C10 : β-cyclodextrin (1:2)

Stability in the solid state complex mesilate of Mitogenome-C10 : β-cyclodextrin (1:2) was studied in the absence of light in three different conditions: 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel according to standard methods of tests of the applicant.

A known weight of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) was placed in a clear glass bottles and stored under different conditions. On two samples taken at 1, 2, 4, 8,16, 32, 64 and 125 day and determine the concentration of the complex mesilate of Mitogenome-C10:β-cyclodextrin method TSRH after dissolution of the samples in water. The results are presented in table 8 and Fig. The results show that mesilate of Mitogenome-C10 stable complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) at 4°C over blue silica gel and at 25°C; 50% relative humidity. At 40°C, 75% relative humidity, 37% of the nelfinavir of Mitogenome-C10 in the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) decomposes during storage over a period of 64 days.

Table 8
Stability in the solid state complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) at 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel
Time (days)1248163264125
4°C over silica gel97,6107,6111,8106,3 106,897,796,899,9
25°C/50% relative humidity96,099,7101,0104,1102,998,198,799,6
40°C/75% relative humidity105,5109,7110,6114,3110,592,065,551,5*

Data represent the average of two values, expressed in percentage of the value at zero time. * means two very different values (71,9 and 31.1%).

Example 10. The study of the stability of the complex mesilate of Mitogenome-C10 : β-cyclodextrin (1:1)

Stability in solution

The stability of the solution of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) define five solvents: water, 0,01M HCl, 0,01M NaOH, IPB (pH of 7.4) and 50% MeOH at two temperatures 25°C and 40°C, in two atmospheric conditions, air and nitrogen for 64 days according to the standard methods of tests of the applicant.

Prepare solutions of complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) (100 mg/ml per mesilate of Mitogenome-C10) in five solvents and diluting the stock solution of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) (1 mg/ml per mesilate of Mitogenome-C10) in the water. Solutions (5 ml) was placed in glass vials, rinsed with air or nitrogen, cover it and put in storage. Selected aliquot amount (0.25 ml) at 0, 1, 2, 4, 8, 16, 32, 64 and 125 day and method TSRH determine concentration.

The results are presented in table 9 and Fig, 27, 28 and 29. The stability of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) in 0,01M NaOH is not included in the data, as mesilate of Mitogenome-C10 decomposes in the solvent for 15 minutes. The results show that (a) the stability of the solution does not depend on the atmosphere above the solution and (b) temperature significantly affects the stability of nelfinavir Mitogenome-C10 in the complex is 1:1 with β-cyclodextrin in water and IPB, but not in the HCl or 50% MeOH.

Table 9
The stability of the solution of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) 4 different solvents under different conditions
ConditionsTime is I (days)
1248163264
Water, air, 25°C101,3of 98.299,599,094,0of 92.789,8
Water, air, 40°C89,487,290,4at 88.189,883,955,2
0,01M HCl, air, 25°C103,0104,3109,4104,6of 99.1102,2100,0
0,01M HCl, air, 40°C94,988,2291,1799,7699,74108,5102,6
IPB, air, 25°C7,9 95,796,1to 97.196,1to 97.195,5
IPB, air, 40°C93,594,199,4105,493,686,975,3
50% MeOH, air, 25°C104,8103,7108,7106,497,696,998,1
50% MeOH, air, 40°C89,385,6891,0193,0br93.192,985,9
Water, N2, 25°C101,7101,3 106,4102,792,092,489,6
Water, N2, 40°C96,291,795,9101,688,084,556,5
0,01M HCl, N2, 25°C103,7106,5108,7to 108.2102,797,2100,3
0,01M HCl, N2, 40°C96,290,997,598,598,5106,7104,8
IPB, N2, 25°C100,199,2to 100.4to 97.196,4of 98.295,5
IPB, N2, 40°C98,4for 95.3102,791,487,975,7
50% MeOH, N2, 25°C101,2101,4104,5102,497,696,499,0
50% MeOH, N2, 40°C94,786,490,086,4to 92.197,487,7

Data represent the average of two values, expressed in percentage of the value at the zero moment of time.

Stability in the solid state

Stability in the solid state complex mesilate of Mitogenome-C10 : β-cyclodextrin (1:1) was studied in the absence of light in three different conditions: 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel according to standard methods of tests of the applicant.

A known weight of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) was placed in a clear glass bottles and stored under different conditions. Two sample taken on 1, 2, 4, 8, 16, 32, 64 and 125 day and determine the concentration of nelfinavir Mit is the quinone-C10 method TSRH after dissolution of the samples in water. The results are presented in table 10 and Fig. The results show that mesilate of Mitogenome-C10 stable at 4°C over blue silica gel and at 25°C; 50% relative humidity, but 37% of the nelfinavir of Mitogenome-C10 in the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) decomposes during storage within 125 days at 40°C, 75% relative humidity.

Table 10
Stability in the solid state complex mesilate of Mitogenome-C10:β-cyclodextrin (1:1) at 40°C, 75% RH; 25°C, 50% RH and 4°C over blue silica gel
Time (days)1248163264125
4°C over silica gel102,197,7100,098,5103,4101,4100,6to 102.3
25°C/50% relative humidity 99,7101,6104,2101,8102,4100,7for 95.2101,9
40°C/75% relative humidityof 98.2101,698,3of 97.898,896,087,266,7

Data represent the average of two values, expressed in percentage of the value at the zero moment of time.

Example 11. Study of the pharmacokinetics of a single IV and oral dose of complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) in rats (P2&P3)

Based on the results of previous pharmacokinetic studies bromide Mitogenome-C10 and study acute oral toxicity of complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2), the dose of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) for the pharmacokinetic study was 50 mg/kg of nelfinavir Mitogenome-C10 for oral dose and 10 mg/kg of nelfinavir Mitogenome-C10 for IV - fold dose.

Ten females of Wistar rats (average weight of approximately 236 g) was Coulibaly tube of Silastic in the right jugular vein using under anesthesia for 48 hours prior to the experiment. Prepared fresh aqueous solution of the complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) (10 mg/ml per mesilate of Mitogenome-C10) and were administered either orally (n=5), or IV by (n=5). Collected blood samples (0.2 ml) at time 0, 5, 10, 20, 30, 45, 60, 90, 120, 180, 240, 360, 720 and 1440 (24 hours) minutes after IV dose and time 0, 15, 30, 60, 90, 120, 150, 180, 240, 300, 420, 540, 720 and 1440 (24 hours) minutes after the oral dose. The blood samples were centrifuged and plasma samples (0.1 ml) were stored at -20°C in the refrigerator. Also collected samples of urine and feces within 24 hours.

The concentration of nelfinavir Mitogenome-C10 - plasma was determined by ESR method using TS/MS (table 12).

Pharmacokinetic analysis

The pharmacokinetics of Mitogenome-C10 analyzed by the method of iterative unweighted nonlinear regression analysis with the least squares using the MINIM. IV data coincided when using one-, two - and three-chamber models. The model that gives the best agreement was the model with the minimum value according to the information criterion of Akaike (A.I.C. Akaike''s information criterion). Found that curves of change of drug concentration in plasma over time after administration of the drug is the best way to adequately correspond to the three open model described by the following equation

C = Ae-αt+Be-βt+Ee-γt

which is predstavljaet the concentration of drug in plasma, A, b and E denote the mathematical coefficients, α denotes a constant velocity distribution in the plasma, β denotes the rate constant of the intermediate phase (distribution or exceptions) and γ denotes the rate constant of the final phase and a slower exception. Half the exception of the drug (t¾ ) in the final phase, was calculated as t¾ =0,693/γ. Data on oral introduction (after 4 h) are consistent with single-chamber model. Concentration peak (Cmaxand the time to reach Cmax(tmax) received directly along the curve of the concentration change in time. The area under the curve of concentration change in time (AUC) was estimated using the linear trapezoid with extrapolation from the last measured concentration to infinity, determined using the rate constants of end-exceptions (γ). General Clarence plasma after intravenous (CL) and oral administration (CL/F) maintenance was estimated as CL = dose/AUC. The volume of distribution was calculated as Vβ= dose / (AUC·β) and Vγ= dose/(AUC·γ). Absolute bioavailability (F) was calculated as F = AUCpox doseiv/AUCivx Dosepo. The mean residence time (MRT - mean residence time was calculated as AUMC/AUC. The apparent volume of distribution at steady state (Vss) was calculated as doseivx AUMC/(AUC)2.

Rezultatai discussion

Curves of changes in the average concentrations of nelfinavir Mitogenome-C10 in the plasma over time after IV and oral administration of complex mesilate of Mitogenome-C10 : β-cyclodextrin (1:2) is shown in Fig, and average pharmacokinetic parameters are listed in table 11. Data on the original content of nelfinavir Mitogenome applied (table 12).

Table 11
Pharmacokinetic parameters of nelfinavir Mitogenome-C10 entered as complex mesilate of Mitogenome-C10:β-cyclodextrin (1:2) to rats after a single IV (10 mg/kg) and oral doses
IV Mitchison-C10 (n=5)Oral dose of Mitogenome-C10 (n=5)
Body weight (g)236,8±21,0236,8±22,9
Cmax(ng/ml)-35,1
tmax(min)-30
t1/2α(min)1,6±0,3-
t1/2β(min)10,4±3,2 -
t1/2γ(h)1,83±0,44-
t1/2(h)14,3*13,9**
AUC (μg·min/ml)47,3±11,129,3±2,7
AUMC (µg·min2/ml)5292±8317477±365
F(%)10012,4
CL (l/min/kg)0,22±0,04-
CL/F (l/min/kg)-13,7±1,3
Vβ(l/kg)3,33±1,46-
Vγ(l/kg)24,04±18,3-
MRT (h)4,2±0,59,5±2,2
Vss (l/kg)25,2±6,5-
* the value of t1/2obtained according to the average concentrations at the time of > 4 hours
** value of t1/ obtained according to the average concentrations at the time of > 4 h

Table 12
The concentration of Mitogenome-C10 in the blood plasma of rats for P2-IV and R3-RO research
Time (min)01 IV (ng/ml)2 IV (ng/ml)3 IV (ng/ml)4 IV (ng/ml)5 IV (ng/ml)Time (min)Average
00,4950,34830,41,391,390
510108671550230016400,11473,4
103063915726414670,2477,2
201991862212511920,3209,8
301321111171581110,5for 125.8
4590,587,57011372,30,886,7
6062,1to 59.4of 56.472,548,71,059,8
9036,4of 37.938,957,429,41,540,0
12023,2to 25.324,2 of 54.818,82,029,3
18017,821,320,730,2223,022,4
24013,112,916,427,49,634,015,9
3607,018,8911,516,77,466,010,3
7202,83,445,664,072,19to 12.03,6
14401,491,631,471,371,9624,01,6
Urine19,352,648,5the 10.112,9
Time (min)06 PO (ng/ml)07 PO (ng/ml)08 PO (ng/ml)09 PO (ng/ml)10 PO (ng/ml)Time (min)Average
0of 0.8782,151,351,390,27901,2
518,3of 21.2of 17.518,344,80,324,0
1029,936,9 26,728540,535,1
201625,419,520,340,81,024,4
3010,7to 25.320,221,1to 25.31,520,5
4524,323,224,525,613,52,022,2
6020,123,825,726,910,42,521,4
902122,419,220,18,723, 18,3
12033,920,91222to 7.674,019,3
18022,417,41313,5the 17.35,016,7
2409,1921,85,035,2421,47,012,5
3606,2720,114,515,115,29,014,2
7208,257,31of 4.384,57between 6.08to 12.06,1
14402,12 0,4182,933,051,7124,02,0
Urine1,5435,44,42br4.6120,7

After IV injection, there is a phase of very rapid distribution, followed by a slower phase distribution or the initial phase of the exception with the subsequent phase of prolonged exclusion at time of approx. 4 o'clock the dependence of the concentration change of Mitogenome-C10 in time satisfies the three models with a finite half-life of 1.8 h, while the half-life, calculated according to the dose after 4 h is 14.3 h (table 13).

After oral administration the absorption of Mitogenome-C10 from the digestive tract of rats was rapid. The peak concentration of Mitogenome-C10 plasma is observed within 1 h after oral administration, and then decreases slowly in time with a half-life of exceptions, calculated on the data after 4 hours, about 14 hours

Set the value of F is 12.4 per cent.

Table 13

Pharmacokinetics of IV (P2) and oral (P3) doses of the complex mesilate of Mitogenome-C10:β-cyclodextrin

All patents, publications, scientific articles and other documents and materials referenced or mentioned in the description, indicate the level of specialists in this field, to which the invention relates, and every such mentioned document and the material included in the description by reference to the extent of fullness, as if it had been included as a separate link or fully described in the specification in its entirety. Applicants reserve the right to physically include in this description any or all materials and information from any such patent publications, scientific articles, web sites, information available through electronic means, and other reference materials or documents.

Specific methods and compositions described in the application are explanatory for the various embodiments or preferred options for implementation and are only examples and are not intended to limit the scope of the claims of the invention. Other objectives, aspects, examples and embodiments of will be clear to experts in this area when considering the present description and are covered by the invention, as defined by the scope of claims of the claims. Specialists in this field it will be understood that various substitutions and modifications can be made in opican the m invention without deviating from the scope of the claims and merits of the invention. The invention illustratively described in the present description, in a suitable embodiment may be carried out in the absence of any element or elements, limitation or limitations, not specifically disclosed in the description as basic. Thus, for example, in each case of embodiments and examples of the present invention any of the terms “comprising”, “consisting essentially of” or “consisting of” may be replaced by any of the other two terms in the description. In addition, the terms “comprising”, “including”, “comprising” and the like should be read in the extended sense and without restrictions. The methods and processes described in this illustrative description, an appropriate image can be carried out in different order stages, and they are not necessarily limited to the orders of the stages specified in them or in the claims. Also used in the description and the attached claims the only form of articles “a”, “an” and “the” include plural references unless the context indicates otherwise. Thus, for example, the term “host cell” includes many (for example, culture or population) of such host cells, and the like, under any circumstances, the patent may not be interpreted as limited to specific examples, implementation options or ways, Conques is to maintain open it. Under no circumstances will the patent may not be interpreted as limited to any claim made by examination or by any other official or employee of the Patent and Trade Office, if such approval is specifically and without qualification or comments made in the response of the applicant.

The terms and expressions which are used, are given in terms of description and not of limitation, and there is no intention to use such terms and expressions to exclude any equivalents of the distinctive features shown or described, or portions thereof, but it is accepted that various modifications are possible within the scope of claims of the invention as it is claimed. Thus, it should be understood that although the present invention specifically disclosed preferred variants of its implementation and possible distinctive features, the experts in this field can have recourse to modifications and changes to the concept disclosed in the description, and such modifications and changes are considered as included in the scope of claims of the present invention, as defined by the attached claims.

The invention described broadly and in General. Each of the shorter fragments and subgeneric groups that fall into the General merits of the invention, also form part of the invention. This includes shared the Scripture of the invention, when the condition or a negative constraint exclusion of any object from the genus, regardless of whether or not the excluded material specifically described.

Other options for implementation are in the scope of the claims claims. In addition, where the distinctive characteristics or aspects of the invention are described in terms of Markush groups, professionals in this field will be clear that the invention is also described in terms of any individual member or subgroup of members of the Markush group.

Compounds of the present invention find application in the selective oxidation treatment of people with the purpose of prevention of mitochondrial damage. This may be a warning of increased mitochondrial oxidative stress associated with specific diseases, such as Parkinson's disease, or diseases associated with mitochondrial DNA mutations. They can also be used for therapy of cell transplant in neurodegenerative diseases, to increase the survival of transplanted cells.

In addition, these compounds can be used as a prophylactic to protect organs during transplantation or improving ischemia-reperfusion injury, which occurs in the process of surgical treatment. Compounds of the present from which retene can also be used to reduce cell damage after a stroke and a heart attack or may be given in the preventive purposes premature children, prone to cerebral ischemia. The methods of the present invention have numerous advantages compared to existing antioxidant therapies they provide the directional accumulation of antioxidants in mitochondria, the part of cells that are under the greatest oxidative stress. This will significantly increase the efficacy of antioxidant therapies.

Specialists in this field will understand that the above description is given only examples and that various combinations of lipophilic cation/antioxidant may be used without deviating from the scope of claims of the invention.

1. Chemically stable antioxidant compound that contains
lipophilic cationic fragment associated connecting with the fragment of the molecule antioxidant; and
anionic component for the specified cationic fragment,
where the antioxidant compound is mitchison selected from
10-(6'-behenoyl)propyltrichlorosilane,
10-(6'-behenoyl)intelligentsia,
10-(6'-behenoyl)deleteperson, and
10-(6'-behenoyl)pentadecatriene having a General formula I

or hinolinol form, where R1, R2and R3represent CH3, atom ()nis saturated and n oznacza is t 3, 5, 10 or 15, and Z means the anion component is chosen from the group consisting of methanesulfonate and econsultant.

2. Chemically stable antioxidant compound containing lipophilic cationic fragment associated connecting with the fragment of the molecule antioxidant and anionic component for the specified cationic fragment, where the antioxidant compound is mitchison, representing 10-(6'-behenoyl)decyltriphenylphosphonium having the formula

and/or its hinolina form, where Z is an anionic component selected from the group consisting of methanesulfonate and econsultant.

3. The compound according to claim 1 or 2, where the anionic component is a methanesulfonate.

4. Chemically stable antioxidant compound containing lipophilic cationic fragment associated connecting with the fragment of the molecule antioxidant and anionic component for the specified cationic fragment, where the antioxidant compound is Mitchison-C10, which is 10-(6'-behenoyl)deleteperson the mesilate, having the formula

and/or its hinolina form.

5. Pharmaceutical composition for reducing oxidative stress in a cell containing an effective amount of chemically stable antioxidant is wow compound according to claim 1 or 2 and a carrier or filler.

6. Pharmaceutical composition for reducing oxidative stress in the cell containing chemically stable antioxidant compound according to claim 4 and a carrier or filler.

7. The pharmaceutical composition according to claim 5, where the anionic component is a methanesulfonate.

8. The pharmaceutical composition according to claim 5, where the antioxidant compound is Mitchison-C10, which is 10-(6'-behenoyl)deleteperson the mesilate, having the formula

and/or its hinolina form.

9. The pharmaceutical composition according to claim 6 or 8, containing β-cyclodextrin.

10. The pharmaceutical composition according to claim 9, where the antioxidant compound and β-cyclodextrin are presented in a molar ratio of compound to cyclodextrin, which is from about 10:1 to about 1:10.

11. The pharmaceutical composition according to claim 9, where the antioxidant compound and β-cyclodextrin are presented in a molar ratio of compound to cyclodextrin, which is selected from the group consisting of (i) from about 5:1 to about 1:5, (ii) from about 4:1 to about 1:4, (iii) from about 2:1 to about 1:2, (iv) about 1:1 and (v) approximately 1:2.

12. The pharmaceutical composition according to claim 9, where the connection and β-cyclodextrin are presented in a molar ratio of connection is the link to the cyclodextrin, which is approximately 1:2.

13. The pharmaceutical composition according to claim 5, which is selected from the group consisting of a pharmaceutical composition formulated for oral administration and the pharmaceutical compositions which are formulated for parenteral administration.

14. The pharmaceutical composition according to claim 9, which contains β-cyclodextrin, and which is chosen from the group consisting of a pharmaceutical composition formulated for oral administration, the pharmaceutical composition formulated for parenteral administration.

15. A method of reducing oxidative stress in the cell, including
contacts mitochondria with chemically stable antioxidant compound according to claims 1, 2 or 4, thereby reducing oxidative stress in the cell.

16. The method according to clause 15, where the antioxidant compound is present in the pharmaceutical composition, which further comprises a carrier or excipient, where the specified media or filler contains β-cyclodextrin.

17. The method according to clause 16, where the antioxidant compound and β-cyclodextrin are presented in a molar ratio of compound to cyclodextrin, which is from about 10:1 to about 1:10.

18. The method according to clause 16, where the connection and β-cyclodextrin are presented in a molar ratio of the compound to cyclodextrin is, which is selected from the group consisting of (i) from about 5:1 to 1:5, (ii) from about 4:1 to 1:4, (iii) from about 2:1 to 1:2, (iv) about 1:1 and (v) approximately 1:2.

19. The method according to clause 16, where the connection and β-cyclodextrin are presented in a molar ratio of compound to cyclodextrin, which is approximately 1:2.

20. The way to obtain antioxidant compounds capable of reducing oxidative stress in a cell, comprising mixing β-cyclodextrin with an antioxidant compound according to any one of claims 1, 2 or 4.

21. The way to obtain antioxidant compounds capable of reducing oxidative stress in a cell, comprising mixing β-cyclodextrin with Makinana-C10, representing 10-(6'-behenoyl)deleteperson mesilate, having the formula

and/or its chinoline form.

22. The method of producing Mitogenome-C10, representing 10-(6'-behenoyl)deleteperson mesilate, having the formula

and/or its hinolina form, including the reaction of debenedetta with triphenylphosphine.

23. The method according to item 22, where idebenone is chemically restored to reaction with triphenylphosphine.

24. The method according to item 22, further comprising prior to the reaction of debenedetta with triphenylphosphine stage
(a) adding t is ethylamine to the solution idebenone to obtain a mixture of idebenone and triethylamine;
(b) cooling the mixture of idebenone and triethylamine stage (a); and
(c) interaction of a mixture of idebenone triethylamine solution methanesulfonanilide to obtain debenedetta.

25. The method according to paragraph 24, which includes at least one of:
(i) stage (a), where the addition of triethylamine provides for the addition of a molar excess of triethylamine in relation to idebenone,
(ii) stage (b), where cooling provides cooling to 10±3°C and
(iii) stage (C), where the interaction involves interaction at about 10-15°C.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a new β-cyclodextrine clathrate complex (an inclusion compound) with 5-hydroxy-4-aminomethyl-1-cyclohexyl(or cycloheptyl)-3-alkoxycarbonylindole derivative: β-cyclodextrine 1:1 to 1:5, preferentially at the relation of 1:1 to 1:3 of general formula (I): wherein X means - hydrogen, chlorine, iodine, n=1 or 2, R3-C1-C3 alkyl, ALK means C1-C6 alkyl group, R1, R2 are independently specified in C1-C4-alkyl, preferentially methyl, or R1 and R2 together with a nitrogen atom (i.e. group - NR1R2) means the groups described by formulas: wherein Bn is benzyl, a Ph is phenyl with the molar ratio of 5-hydroxy-4-aminomethyl-1-cyclohexyl(or cycloheptyl)-3-alkoxycarbonylindole derivative: β-cyclodextrine 1:1 to 1:5, preferentially 1:1 to 1:3, especially preferentially in the relation of 1:2. The clathrate complex may represent nanoparticles of size not less than 100 nm. There are preferential clathrate complexes wherein 5-hydroxy-4-aminomethyl-1-cyclohexyl(or cycloheptyl)-3-alkoxycarbonylindole derivative represents 1-cyclohexyl-4-aminomethyl-5-hydroxy-2-methyl-1H-indole-3-carboxylic acid ethyl ester. The new clathrate complexes possess antiviral action and exhibit high activity versus influenza viruses. The invention also involves a pharmaceutical composition and a drug based on the clathrate complexes. Besides, the invention refers to liquid-phase and solid-phase synthesis of the clathrate complexes.

EFFECT: preparing the compounds which possess antiviral action and exhibit high activity versus influenza viruses.

20 cl, 2 ex, 2 tbl, 8 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to a novel clathrate complex of β-cyclodextrin with 1-{[6-bromo-1-methyl-5-methoxy-2-phehylthiomethyl-1-H-indol-3-yl]carbonyl}-4-benzylpiperazine of formula : with molar ratio 1-{[6-bromo-1-methyl-5-methoxy-2-phehylthiomethyl-1-H-indol-3-yl]carbonyl}-4-benzylpiperazine: β-cyclodextrin from 1:1 to 1:10, synthesis method and use thereof as an antiviral agent for treating influenza. The disclosed method involves mixing solutions of β-cyclodextrin and 1-{[6-bromo-1-methyl-5-methoxy-2-phehylthiomethyl-1-H-indol-3-yl]carbonyl}-4-benzylpiperazine in molar ratio from 1:1 to 1:10 while stirring and heating to temperature not higher than 70°C and then maintaining said conditions until a homogeneous solution is obtained and extraction of the obtained complex.

EFFECT: clathrate complex is a novel effective anti-influenza virus agent which is obtained using a novel efficient method.

13 cl, 2 ex, 3 tbl, 11 dwg

FIELD: chemistry.

SUBSTANCE: method of intensifying antiaggregant activity in an experiment involves use of a chemical compound which is a conjugate of beta-cyclodextrin with acetylsalicylic acid.

EFFECT: wider range of agents for increasing antiaggregant activity.

1 tbl

FIELD: chemistry.

SUBSTANCE: method of obtaining complex with inclusion of cyclodextrin can include dry mixing of cyclodextrin and hydrocolloid for formation of dry mixture and mixing solvent and guest with dry mixture for formation of complex with inclusion of cyclodextrin. In some versions of realisation method of obtaining complex with inclusion of cyclodextrin can include mixing of cyclodextrin and hydrocolloid for formation of first mixture, mixing of first mixture with solvent for formation of second mixture and mixing of quest with second mixture for formation of third mixture.

EFFECT: elaboration of efficient method of obtaining complex with inclusion of cyclodextrin.

41 cl, 17 ex

FIELD: chemistry.

SUBSTANCE: molar ratio of cyclodextrin to the acid in the complex comprises 1:1. The complex is obtained by introduction of concentrated water cyclodextrin solution heated to boiling point to glacial acetic acid of room temperature, with further separation and drying of crystalline sediment. Complex of α- or β-cyclodextrin with acetic acid is stable in dry state, but in water solution it is decomposed into components. Obtained solution gains properties of dilated acetic acid, and therefore can be used as flavouring in food concentrates, as preservation agent, solution acidity regulator, and as buffer component in biochemistry and analytical chemistry.

EFFECT: enhanced efficiency of composition.

3 cl, 4 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to cyclodextrin-containing polymeric compounds, which are carriers for delivery of therapeutics, and pharmaceutical preparations based on them. Invention also relates to method of treating subjects with therapeutically effective quantity of said cyclodextrin-containing polymeric compound. Claimed cyclodextrin-containing polymers improve medication stability, increase its solubility and reduce toxicity of therapeutics when used in vivo. Furthermore, by selecting from a variety of linker groups and targeting ligands of said polymers it is possible to realise controlled delivery of therapeutic agents.

EFFECT: obtaining cyclodextrin-containing polymer compounds, improving medication stability, increasing its solubility and reducing toxicity of therapeutics when used in vivo.

56 cl, 13 dwg, 7 tbl, 46 ex

FIELD: chemical technology.

SUBSTANCE: invention relates to the inclusion complex of cyclodextrins with elemental sulfur. Complex can be prepared using different homologues of cyclodextrins, for example, beta- and gamma-cyclodextrins and hydroxypropylated forms of gamma- and beta-cyclodextrins. Proposed complex can be used as a biologically active compound for medicinal, veterinary and agricultural designation. Invention provides the possibility for further preparing true solutions of elemental sulfur in water in the concentration up to 250-300 mg/l.

EFFECT: improved preparing method, valuable properties of complex.

7 cl, 7 ex

FIELD: chemical technology.

SUBSTANCE: invention describes a method for preparing immobilized β-cyclodextrin. Method involves pretreatment of organic and inorganic sorbents - polyvinyl alcohol and sorbitol based on silica with glutaraldehyde for incorporation of aldehyde group molecules into sorbents, washing out with water and dimethylsulfoxide on glass porous filter, drying on glass porous filter followed by addition of prepared sorbents to dimethylsulfoxide solution containing dissolved β-cyclodextrin in the ratio sorbent : β-cyclodextrin : dimethylsulfoxide = 1.0:(0.4- 2.0);10, respectively, stirring the prepared suspension at temperature 25-70°C for 30-180 min, washing out and drying. Method provides preparing an insoluble sorbent with immobilized β-cyclodextrin used for removing cholesterol or its derivatives.

EFFECT: improved preparing method.

5 ex

FIELD: organic chemistry, medicine.

SUBSTANCE: invention describes a derivative of 6-mercaptocyclodextrin of the general formula (I):

wherein n = 0-7; n = 1-8 and m + n = 7 or 8; R represents (C1-C6)-alkylene substituted optionally with 1-3 OH-groups, or (CH2)o-phenylene-(CH2)p wherein o and p = 0-4 independently; X represents COOH, CONHR1, NHCOR2, SO2OH, PO(OH)2, O(CH2-CH2-O)q-H, OH or tetrazole-5-yl; R1 represents hydrogen atom (H) or (C1-C3)-alkyl; R2 represents carboxyphenyl; q = 1-3; or its pharmaceutically acceptable salt in mixture with pharmaceutically acceptable accessory substances. Also, invention describes a set and pharmaceutical composition for reversing drug-induced neuromuscular blocking comprising derivative of 6-mercaptocyclodextrin of the general formula (I), and a method for reversing drug-induced neuromuscular blockade in patient that involves parenteral administration to indicated patient the effective dose of 6-mercaptocyclodextrine derivative of the general formula (I) by cl. 1.

EFFECT: valuable medicinal properties of agents.

11 cl, 1 tbl, 20 ex

The invention relates to a linear cyclodextrin copolymers and oxidized cyclodextrin, which can be used as a carrier for the delivery of various therapeutic agents

FIELD: medicine.

SUBSTANCE: invention refers to phosphonium salt of pyridoxine derivatives of general formula (I) which may be used in medicine and veterinary science wherein if R1=CH3; R2=CH3; X=H; n=1; R1=CH3; R2=H; X-H; n=1; R1=CH3; R2=CH3; X-CH2P*Ph3; n=2; R,=CH3; R2=H; X=CH2P+Ph3; n=2; R,=CH3; R2=C(CH3)2; X=CH2P+Ph3; n=2.

EFFECT: there are presented new compounds with antibacterial activity on Staphylococcus aureus, including multiresistant strains, as well as low toxicity.

1 cl, 1 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula I, production and use thereof to obtain corresponding organophosphinates [Kt]z+ z[(CnHmF2n+1-m)xPCIyF6.x.y]- (I) where [Kt]z+ is an organic cation of formula (1) [NR4]' (1) where R is an optionally phenyl-substituted linear C1-4-alkyl; formula (2) [PR24]+ (2) where R2 is independently C6-14-alkyl; or formula (6) [HetN]z+ (6), where HetNz+ is a heterocyclic cation selected from a group comprising imidazolium, pyrazolium, dihydroimidazolium, pyrrolidinium, triazolium, pyridinium, pyridazinium, pyrimidinium, piperidinium, piperazinium, pyrazinium, R1,-R4, denote H or C1-10-alkyl; n=1-4, m=0 to 2n+1, x=1-4, y=1, z=1-2, under the condition that x+y<5.

EFFECT: novel compounds, a method of producing said compounds and use of said compounds to obtain valuable compounds are disclosed.

12 cl, 10 ex

FIELD: chemistry.

SUBSTANCE: method involves treating a water-based medium containing sulphate-reducing bacteria SRB in industrial aqueous systems of chemical production and oil refining. Inhibition of production of biogenic sulphide with SRB takes place as a result of synergetic action of a biocide component in a first concentration and a metabolic inhibitor in a second concentration. The biocide immediately destroys the first portion of SRB. The biocide component is selected from a group comprising aldehydes, amine-type compounds, halogenated compounds, sulphur compounds, salts of quaternary phosphonium and/or combinations thereof. The metabolic inhibitor inhibits growth of a second portion of SRB without its direct destruction. The metabolic inhibitor component is selected from a group comprising nitrite, molybdate, tungstate, selenate, anthraquinone and/or combinations thereof. Contact between the SRB and the biocide and metabolic inhibitor can take place continuously, intermittently or simultaneously.

EFFECT: method ensures efficient inhibition of production of biogenic sulphide with SRB during combined use of components in considerably lower concentrations than if the biocide or metabolic inhibitor was used separately.

25 cl, 2 dwg, 1 ex

FIELD: chemistry.

SUBSTANCE: present invention relates to 2-(carboxy-n-alkyl)ethyltriphenyl phosphonium bromides of general formula I, having bactericidal and fungicidal activity, heat resistance and surfactant resistance, which can be used in veterinary, medicine and agriculture , where R = n-C10H21 n-C12H25, n-C14H29, n-C16H33, n-C18H37.

EFFECT: obtaining novel biologically active compounds.

1 cl, 6 ex, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to a new improved method of producing onium tetrafluoroborates through reaction of an onium halide with trialkyloxonium tetrafluoroborate, trialkylsulphonium tetrafluoroborate or triphenylcarbonium tetrafluoroborate, characterised by that the halide has formula (1) [XR4]+ Hal-, where X denotes N, P, Hal denotes Cl, Br or I and R in each case independently denotes a linear alkyl having 1-8 C atoms, or the halide has formula (2) [(R1R2N)-C(=SR7)(NR3R4)]+ Hal- (2), where Hal denotes Br or I R1-R7 each independently denotes a linear alkyl having 1-8 C atoms, or the halide has formula (3) [C(NR1R2)(NR3R4)(NR5R6)]+ Hal- (3), where Hal denotes CI, Br or I and R1-R6 each independently denotes a linear alkyl having 1-8 C atoms, or the halide has formula (4) [HetN]+ Hal- , where Hal denotes CI, Br or I and HetN+ denotes a heterocyclic cation selected from a group comprising imidazolium pyrrolidinium pyridinium where each of substitutes R1' - R4' independently denotes hydrogen, CN, linear or branched alkyl having 1-8 C atoms, dialkylamine containing alkyl groups having 1-4 C atoms but which is not attached to he heteroatom of the heterocyclic ring.

EFFECT: method enables to obtain products with low content of halides with high purity and high output.

5 cl, 12 ex

FIELD: chemistry.

SUBSTANCE: invention relates to ionic liquid used in electrical energy accumulation devices and as a solvent which contains a cation of general formula where X1, X2 and X3 denote N, O, S or C; R1-R11, X1, R1, R2 and R3, X2, R6, R7 and R8, X3, R9, R10 and R11 can form ring structures; the anion is selected from [RSO3]-, [RfSO3]-, [(RfSO2)2N]-, [(RfSO2)3C]-, [(FSO2)3C]-, [ROSO3]-, [RC(O)O]-, [RfC(O)O]-, [CCl3C(O)O]-, [(CN)3C]-, [(CN)2CR]-, [(RO(O)C)2CR]-, [R2P(O)O]-, [RP(O)O2]2-, [(RO)2P(O)O]-, [(RO)P(O)O2]2-, [(RO)(R)P(O)O]-, [Rf2P(O)O]-, [RfP(O)O2]2-, [B(OR)4]-, [N(CN)2]-, [AlCl4]-, PF6-, [RfPF5]-, BF4-, [RfBF3]-, SO42-, HSO4-, NO3- I-, bis(oxalate)borate; R, R1-R11 are selected from hydrogehn, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and heterocyclyl, halogen, CN- or NO2-; the carbon in R and R1-R11 can be substituted with O-, -Si(R')2-, -C(O)-, -C(O)O-, -S-, -S(O)-, -SO2-, -SO3-, -N= -N=N-, -NH-, -NR'-, -N(R')2-, -PR'-, -P(O)R4 -P(O)R'-O-, -O-P(O)R'-O- and -P(R')2=N-; where R' denotes alkyl, fluoroalkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, phenyl or heterocyclyl; Rf denotes a fluorine-containing substitute.

EFFECT: obtaining novel ionic liquids which are stable in liquid state in a wide temperature range.

14 cl, 76 ex, 3 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to ionic liquid used in electrical energy accumulation devices and as a solvent which contains a cation of general formula where X1, X2 and X3 denote N, O, S or C; R1-R11, X1, R1, R2 and R3, X2, R6, R7 and R8, X3, R9, R10 and R11 can form ring structures; the anion is selected from [RSO3]-, [RfSO3]-, [(RfSO2)2N]-, [(RfSO2)3C]-, [(FSO2)3C]-, [ROSO3]-, [RC(O)O]-, [RfC(O)O]-, [CCl3C(O)O]-, [(CN)3C]-, [(CN)2CR]-, [(RO(O)C)2CR]-, [R2P(O)O]-, [RP(O)O2]2-, [(RO)2P(O)O]-, [(RO)P(O)O2]2-, [(RO)(R)P(O)O]-, [Rf2P(O)O]-, [RfP(O)O2]2-, [B(OR)4]-, [N(CN)2]-, [AlCl4]-, PF6-, [RfPF5]-, BF4-, [RfBF3]-, SO42-, HSO4-, NO3- I-, bis(oxalate)borate; R, R1-R11 are selected from hydrogehn, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and heterocyclyl, halogen, CN- or NO2-; the carbon in R and R1-R11 can be substituted with O-, -Si(R')2-, -C(O)-, -C(O)O-, -S-, -S(O)-, -SO2-, -SO3-, -N= -N=N-, -NH-, -NR'-, -N(R')2-, -PR'-, -P(O)R4 -P(O)R'-O-, -O-P(O)R'-O- and -P(R')2=N-; where R' denotes alkyl, fluoroalkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, phenyl or heterocyclyl; Rf denotes a fluorine-containing substitute.

EFFECT: obtaining novel ionic liquids which are stable in liquid state in a wide temperature range.

14 cl, 76 ex, 3 dwg

FIELD: chemistry.

SUBSTANCE: present invention relates to ionic liquids based on a cation of formula (1): where substituting groups R1-R9 are selected from hydrogen, alkyl; any carbon atom in R1-R9 can be substituted with a -O-, -C(O)-, -C(O)O-, -S-, -S(O)-, -SO2- or -SO3- group; X is S, O or C; R8 and R9 exist only when X is carbon; the anion is selected from [RSO3]-, [RfSO3]-, [(RfSO2)2N]-, [(FSO2)3C]-, [RCH2OSO3]-, [RC(O)O]-, [RfC(O)O]-, [CCl3C(O)O]-, [(CN)3C]-, [(CN)2CR]-, [(RO(O)C)2CR]-, [B(OR)4]-, [N(CF3)2]-, [N(CN)2]-, [AlCl4]-, PF6-, BF4-, SO42-, HSO4-, NO3-; where R is hydrogen, halogen, alkyl, alkenyl, alkynyl, cycloalkyl, Rf is a fluorine-containing substituting group.

EFFECT: obtaining new ionic liquids with improved electrochemical properties.

15 cl, 18 ex, 2 dwg

FIELD: chemistry.

SUBSTANCE: claimed invention relates to copolymers of diallylaminophosphonium salts with sulphur dioxide demonstrating antimicrobial activity with respect to a number of bacteria, as well as to yeast-like fungi and spores, and can be applied as antiseptic and disinfecting means. Claimed copolymers of diallylaminophosphonium salts with sulphur dioxide aree characterised by general formula where A=Cl- or BF4-. They are soluble in methanol, DMSO, DMFA or if A=Cl- are soluble in water. They are obtained by copolymerisation of equimolar amounts of sulphur dioxide and diallylaminophosphonium salt, selected from tris(diethylamino)diallylaminophosphonium chloride or tris(diethylamino)diallylaminophosphonium tetrafluoroborate.

EFFECT: obtaining novel efficient and low-toxic compounds which do not cause corrosion of processed metals.

2 cl, 3 tbl, 4 ex

FIELD: chemistry.

SUBSTANCE: present invention concerns the salts containing bis(trifluoromethyl)imide anions and saturated, partially or completely unsaturated heterocyclic cations, method of production and application thereof as ionic liquids.

EFFECT: production of new salts to be used as ionic liquids.

19 cl, 5 ex

FIELD: organic chemistry, analytical chemistry.

SUBSTANCE: method for concentrating hydroquinone from aqueous solutions involves extraction with an organic solvent wherein tributyl phosphate is used as an organic solvent that is applied preliminary of foam polyurethane in the mass ratio polyurethane foam : tributyl phosphate = 1:(2.5-3.0). Proposed method provides: to enhance the concentrating coefficient by 2.5-fold; to enhance extraction degree up to 96%; to exclude desalting agent from extraction system, and to reduce cost of analysis. Invention can be recommended for concentrating hydroquinone in analytical control of sewage feeding to biological treatment.

EFFECT: improved concentrating method.

1 tbl

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