Abstract:
Provided are stable liquid and solid formulations of oxidized and reduced mitochondria-targeted antioxidants, and methods of their preparation and use.

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This application is the national phase under 35 U.S.C. §371 of International Application No. PCT/US2012/040711, entitled “Oral Formulations of Mitochondrially-Targeted Antioxidants and Their Preparation and Use,” filed on Jun. 4, 2012, which claims priority to and the benefit of U.S. Provisional Patent application Ser. No. 61/492,940 entitled “Oral Formulations of Mitochondrially-Targeted Antioxidants and Their Medical Use” which was filed Jun. 3, 2011. The entirety of the aforementioned applications are herein incorporated by reference. 
    
    
     FIELD OF THE INVENTION 
     This disclosure is in the fields of cell biology, pharmacology and medicine, and in particular, inflammation, diabetes, septic shock, wound healing, and coronary heart disease. 
     BACKGROUND 
     Promising therapeutical properties of mitochondria-targeted antioxidants (MTAs) have been described (see, e.g., US2008176929; Skulachev et al. (2009),  Biochim. Biophys. Acta,  1787:437-61). The experiments performed which revealed these properties were done with freshly prepared solutions of MTAs and made by dissolving of ethanol stock solutions preserved at −80° C. shortly before administration of the preparation to animals. Such method of preparation and administration is not suitable or realistic for preparation of pharmaceuticals as it is extremely inconvenient if not impossible for industrial manufacturing, logistics, and use by patients. Attempts to develop a pharmaceutical composition (for oral administration or injection) with acceptable stability revealed that MTAs are not stable in most types of oral or injectable compositions. Stable pharmaceutical composition containing MTAs possessing acceptable stability have not been described up to now. Accordingly, improved liquid formulations with stability are still needed. 
     SUMMARY 
     The present disclosure provides stabilized liquid and solid formulations comprising MTAs suitable for oral, nasal, and intravenous and injectable administration, and methods of preparation of such formulations. The invention also provides methods of treatment and prophylaxis of diseases and conditions relating to mitochondria using such formulations. 
     In one aspect, the disclosure provides a stabilized pharmaceutical formulation comprising a compound of Formula I in oxidized and/or reduced form. 
     The compound of Formula I is: 
                                
wherein:
 
     A is an antioxidant of Formula II: 
                                
and/or reduced form thereof, wherein m comprises an integer from 1 to 3;
 
     Y is independently selected from the group consisting of: lower alkyl, lower alkoxy, or two adjacent Y groups, together with carbon atoms to which they are attached, form a following structure of Formula III: 
                                
and/or reduced form thereof, wherein:
 
     R1 and R2 are the same or different and are each independently lower alkyl or lower alkoxy; 
     L is a linker group, comprising: a) a straight or branched hydrocarbon chain optionally substituted by one or more double or triple bond, or ether bond, or ester bond, or C—S, or S—S, or peptide bond; and which is optionally substituted by one or more substituents preferably selected from alkyl, alkoxy, halogen, keto group, amino group; or b) a natural isoprene chain; 
     n is an integer from 1 to 20; and 
     B is a targeting group comprising: a) a Skulachev-ion Sk (Sk + Z − ) wherein: Sk is a lipophillic cation or a lipophillic metalloporphyrin, and Z is a pharmaceutically acceptable anion; or b) an amphiphillic zwitterion, 
     with the proviso that in compound of Formula I, A is not ubiquinone (e.g., 2-methyl-4,5-dimethoxy-3,6-dioxo-1,4-cyclohexadienyl) or tocopherol or a mimetic of superoxide dismutase or ebselen; when L is divalent decyl, divalent pentyl, or divalent propyl radical; and when B is triphenylphosphonium cation. 
     In a particular embodiment, the composition is reduced or is oxidized. In some embodiments, the formulation is in liquid form, and in other embodiments, the formulation is in solid form. 
     In some embodiments the liquid formulation comprises a compound of Formula I in 10% to 100% glycerol, from about 10% to about 100% glycol, (e.g., 1,2-propylene glycol) or from about 1% to about 100% (absolute) ethanol. In one particular embodiment, the composition of Formula I is in about 50% 1,2-propylene glycol. 
     The disclosure also provides stabilized solid pharmaceutical formulations comprising a compound of Formula I in oxidized or reduced form, with the proviso that in compound of Formula I, A is not ubiquinone (e.g., 2-methyl-4,5-dimethoxy-3,6-dioxo-1,4-cyclohexadienyl) or tocopherol or a mimetic of superoxide dismutase or ebselen; when L is divalent decyl, divalent pentyl, or divalent propyl radical; and when B is triphenylphosphonium cation. 
     In one embodiment, the formulation also comprises 1 molar equivalent to 200 molar equivalents of an antioxidation agent that reduces the oxidized form of the compound of Formula 1, and a pharmaceutically acceptable carrier. 
     In some embodiments, the antioxidation agent is ascorbic acid. 
     In some embodiments, the pharmaceutically acceptable carrier comprises sorbite, glucose, and/or magnesium stearate. 
     In certain embodiments, the pharmaceutical formulation is SkQ1 or SkQ1H 2 . In other embodiments, the compound is SkQR1 or SkQR1H 2 . In yet other embodiments, the compound is SkQ3 or SkQ3H 2 . In still other embodiments, the compound is SkQRB or SkQRBH 2 . In other embodiments, the compound is SkQB1 or SkQB1H 2 . In yet other embodiments, the compound is SkQBP1 or SkQBP1H 2 . 
     In other aspects, the disclosure provides methods of treating and preventing diabetes type I and II, inflammation, septic shock, arthritis, and coronary heart disease, and methods of aiding in wound healing. In these methods, a therapeutically effective amount of a formulation comprising a stabilized compound of Formula I in liquid or solid form is administered to a patient, with the proviso that in compound of Formula I, A is not ubiquinone (e.g., 2-methyl-4,5-dimethoxy-3,6-dioxo-1,4-cyclohexadienyl) or tocopherol or a mimetic of superoxide dismutase or ebselen; when L is divalent decyl, divalent pentyl, or divalent propyl radical, and when B is triphenylphosphonium cation. 
     In some embodiments of the method, the formulation comprises glycerol, glycol, and/or ethanol. In some embodiments, the formulation comprises SkQ1, SkQ1H 2 , SkQR1, SkQR1H 2 , SkQ3, SkQ3H 2 , SkQBP1, SkQBP1H 2 , SkQRB, or SkQRBH 2 . 
     In some embodiments, the liquid formulation is administered orally or by injection. In other embodiments, the solid formulation is administered orally, anally, or vaginally. In some embodiments the formulation is a solid and comprises ascorbic acid. In particular embodiments, the formulation also comprises a pharmaceutically acceptable carrier. 
     In some embodiments, diabetes type I or II is treated with SkQ1 or SkQ1H 2  in 20% glycerol. 
     In certain embodiments, arthritis is treated with a formulation comprising SkQ1 or SkQ1H 2  in 20% glycerol. In yet other embodiments, arthritis is treated with a formulation comprising SkQ1 and ascorbic acid. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The foregoing and other objects of the present disclosure, the various features thereof, as well as the invention itself may be more fully understood from the following description, when read together with the accompanying drawings. 
         FIG. 1  is a graphic representation of the effect of SkQ1 on blood glucose level of diabetic animal model (alloxan-treated mice); 
         FIG. 2  is a graphic representation of the effect of SkQ1 on liver damage of db/db diabetic mice; 
         FIG. 3   a  is a graphic representation illustrating the effect of SkQ1 on epithelization of diabetic wounds; 
         FIG. 3   b  is a graphic representation illustrating the effect of SkQ1 on the amount of neutrophils in diabetic wounds; 
         FIG. 3   c  is a graphic representation illustrating the effect of SkQ1 on vessel density in diabetic wounds; 
         FIG. 4  is a graphic representation of the effect of SkQ1 on survival of mice subjected to septic shock; 
         FIG. 5  is a graphic representation demonstrating the anti-inflammatory effect of SkQ1 in collagen-induced arthritis in rats; 
         FIG. 6  is a graphic representation demonstrating the anti-inflammatory effect of SkQ1 and SkQR1 rescuing endothelial cells from death induced by proinflammatory cytokine TNF-alpha; 
         FIG. 7   a  is a graphic representation demonstrating the ability of SkQ1 to inhibit inflammation in vitro by lowering expression of pro-inflammatory cytokines; and 
         FIG. 7   b  is a graphic representation demonstrating the ability of SkQ1 to inhibit inflammation in vivo by lowering expression of pro-inflammatory cytokines as measured by relative ICAM-1 mRNA expression in mice. 
     
    
    
     DESCRIPTION 
     Throughout the text of a description of the invention various documents are cited. Each document cited here (including all patents, patent applications, scientific publications, specifications and manufacturer&#39;s instructions etc.), above or below, is introduced in full in this invention by reference. 
     Prior to the detailed description of the invention follows, one should understand that the invention is not limited to the particular methodology, protocols, and reagents described here, as they are subject to change. In addition, it should be understood that in the present invention, the terminology is used to describe particular embodiments only and does not limit the scope of the present invention which will be limited only by the appended claims. Unless otherwise specified, all technical and scientific terms used here have the same meanings that are understandable to those skilled in the art. 
     It was unexpectedly found that many effective MTAs are not stable enough in usual liquid and solid pharmaceutical formulations suitable for their administration by injection, or by oral, IV, nasal, topical, or enteral administration. This feature limits clinical application of pharmaceuticals based on MTA as active compounds. 
     I. Stabilized Formulations 
     The present disclosure provides stable, liquid, MTA-based pharmaceutical compositions applicable in clinical practice. A useful MTA is a compound of Formula I in oxidized and/or reduced form. 
     The compound of Formula I is: 
                                
wherein:
 
     A is an antioxidant of Formula II: 
                                
and/or reduced form thereof, wherein m comprises an integer from 1 to 3;
 
     Y is independently selected from the group consisting of: lower alkyl, lower alkoxy, or two adjacent Y groups, together with carbon atoms to which they are attached, form a following structure of Formula III: 
                                
and/or reduced form thereof, wherein:
 
     R1 and R2 are the same or different and are each independently lower alkyl or lower alkoxy; 
     L is a linker group, comprising: a) a straight or branched hydrocarbon chain optionally substituted by one or more double or triple bond, or ether bond, or ester bond, or C—S, or S—S, or peptide bond; and which is optionally substituted by one or more substituents preferably selected from alkyl, alkoxy, halogen, keto group, amino group; or b) a natural isoprene chain; 
     n is an integer from 1 to 20; and 
     B is a targeting group comprising: a) a Skulachev-ion Sk: (Sk + Z − ), wherein: Sk is a lipophillic cation or a lipophillic metalloporphyrin, and Z is a pharmaceutically acceptable anion; or b) an amphiphillic zwitterion, with the proviso that in compound of Formula I, A is not ubiquinone (e.g., 2-methyl-4,5-dimethoxy-3,6-dioxo-1,4-cyclohexadienyl) or tocopherol or a mimetic of superoxide dismutase or ebselen; when L is divalent decyl, divalent pentyl, or divalent propyl radical; and when B is triphenylphosphonium cation, with the proviso that in compound of Formula I, A is not ubiquinone (e.g., 2-methyl-4,5-dimethoxy-3,6-dioxo-1,4-cyclohexadienyl) or tocopherol or a mimetic of superoxide dismutase or ebselen; when L is divalent decyl, divalent pentyl, or divalent propyl radical; and when B is triphenylphosphonium cation. 
     Specific useful MTAs include, but are not limited to, the SkQ1 and SkQR1: 
                                
and their reduced (quinole) forms SkQ1H 2  and SkQR1H 2 , respectively. These MTAs have been described in PCT/RU2006/000394.
 
     Other useful MTA variants include, but are not limited to SkQ3: 
                                
and its reduced (quinole) form SkQ3H 2 ;
 
     to SkQRB: 
                                
and its oxydized (quinone) form SkQRB;
 
     to SkQB1: 
                                
and its reduced (quinole) form, SkQB1H 2 ; and
 
     to SkQBP1: 
                                
and its reduced (quinole) form SkQBP1H 2 .
 
     These MTAs are formulated for oral administration as liquid solutions and as solid formulations. 
     Liquid solutions are also useful for aerosol delivery via injection, for IV administration, nasal administration, topical administration, or enteral administration. 
     Such stable liquid formulations include one or more solvents or soluble components into which the MTAs are placed. Useful solvents include glycerol, ethanol, propyleneglycol, and analogous compounds. For example, useful stable formulations contain at least 10% 1,2-propylene glycol, at least 1% or at least 10% ethanol, at least 10% glycerol, or mixtures thereof, which may also include water, glycerol, ethanol, and/or 1,2-propylene to make up the difference. For example, representative stabilizing solutions of 1 nM to 1 mM SkQ1, SkQ1H 2 , SkQR1, SkQR1H 2 , SKQ3, SkQ3H 2 , SKQRB, SkQRBH 2 , SKQB1, SkQB1H 2 , SKQBP1 and/or SkQBP1H 2 , contain 10% to 50%, 50% to 100%, 10% to 20%, 20% to 30%, 30% to 40%, 40% to 50%, 50% to 60%, 60% to 70%, 70% to 80%, 80% to 90%, 10% to 100%, 20% to 80%, and 90% to 100% 1,2-propylene glycol, glycerol, or ethanol. Other useful percentages of such solvents include 15%, 20%, 25%, 30%, 35%, 40%, 45%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, and 95%. Other pharmaceutically acceptable carriers may also be components of such formulations. 
     Because MTAs are not shelf-stable for long periods of time, various compounds were tested to determine their ability to stabilize SkQ1 and SkQR1 as representative MTAs in dry form. 
     Beta-cyclodextrin, gun-arabic, fruit fibers, and sodium chloride did not provide suitable stabilization levels (degradation rate, %/d was 0.8 to 8.1). 
     Liquid solvents were also tested for their ability to stabilize representative MTAs SkQ1 and SkQR1. The solvents tested were water solutions of glycerol (10% to 100%), 50% lactulose, and 1,2-propylene glycol (10% to 100%, at 60° C.). Some representative results are shown below (Table 1). 
     
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                   
                   
                   
                 Degradation 
               
               
                   
                 Concentration, 
                 Stabilizing 
                 rate, percent 
               
               
                 MTA 
                 mkM 
                 Solvent 
                 per day 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 SkQ1 
                 400 
                  50% lactulose 
                 9.01 
               
               
                 SkQ1 
                 400 
                  10% 1,2-propylene glycol 
                 0.47 
               
               
                 SkQ1 
                 400 
                  50% 1,2-propylene glycol 
                 0.06 
               
               
                 SkQ1 
                 400 
                 100% 1,2-propylene glycol 
                 0.18 
               
               
                 SkQ1 
                 400 
                  10% Glycerol 
                 0.61 
               
               
                 SkQ1 
                 400 
                  20% Glycerol 
                 0.51 
               
               
                 SkQ1 
                 400 
                  30% Glycerol 
                 0.53 
               
               
                 SkQ1 
                 400 
                  40% Glycerol 
                 0.91 
               
               
                 SkQ1 
                 400 
                  50% Glycerol 
                 1.54 
               
               
                 SkQ1 
                 400 
                  60% Glycerol 
                 1.92 
               
               
                 SkQ1 
                 400 
                  70% Glycerol 
                 2.4 
               
               
                 SkQ1 
                 400 
                  80% Glycerol 
                 3.2 
               
               
                 SkQ1 
                 400 
                  90% Glycerol 
                 4.18 
               
               
                 SkQRB 
                 200 
                  50% Glycerol 
                 0.4 
               
               
                 SkQR1 
                 140 
                  50% Glycerol 
                 0.7 
               
               
                 SkQBP1 
                 400 
                  50% Glycerol 
                 0.08 
               
               
                 SkQR1 
                 100 
                  10% 1,2-propylene glycol 
                 6.19 
               
               
                 SkQR1 
                 100 
                  20% 1,2-propylene glycol 
                 0.34 
               
               
                 SkQR1 
                 100 
                  30% 1,2-propylene glycol 
                 0.32 
               
               
                 SkQR1 
                 100 
                  40% 1,2-propylene glycol 
                 0.06 
               
               
                 SkQR1 
                 100 
                  50% 1,2-propylene glycol 
                 &lt;0.01 
               
               
                 SkQR1 
                 100 
                  60% 1,2-propylene glycol 
                 &lt;0.01 
               
               
                 SkQR1 
                 100 
                  70% 1,2-propylene glycol 
                 0.05 
               
               
                 SkQR1 
                 100 
                  80% 1,2-propylene glycol 
                 0.23 
               
               
                 SkQR1 
                 100 
                  90% 1,2-propylene glycol 
                 0.30 
               
               
                 SkQR1 
                 100 
                 100% 1,2-propylene glycol 
                 0.23 
               
               
                   
               
             
          
         
       
     
     These results illustrate high stability of MTAs in a pharmaceutical composition for administration in the form of solution in glycerol (from about 10% to about 100% glycerol), and about 50% 1,2-propylene glycol solution. 
     In addition, the stability of SkQ1 and SkQR1 was significantly increased in dark plastic or glass vials, indicating that these compounds are light-sensitive. Accordingly, one of the ways to further improve or increase stability of SkQ liquid compositions during storage and transportation is to protect it from light. 
     When SkQ compounds of Formula I according to the disclosure are in solid form, they may be stabilized, for example, with an antioxidation agent. Such an agent can be ascorbic acid. Useful amounts of ascorbic acid range from about 1 molar equivalent to about 200 molar equivalents. As used herein, the term “molar equivalent” refers to the number of dissolved particles, or that amount which reacts with, or supplies one mole of H +  in an acid-base reaction, or which reacts or supplies one mole of electrons in a redox reaction. Other useful components of representative stabilized MTA formulations are shown in Table 2. Such formulations may also comprise pharmaceutically acceptable carriers such as, but not limited to, sorbite, glucose, and magnesium stearate. 
     Another approach to stabilize an SkQ compound in a pharmaceutical formulation is to use its reduced (quinole) form. For example, the reduced form of SkQ1 is the quinole SkQ1H 2 : 
                                
where Z −  is pharmaceutically acceptable anion such as, but not limited to, bromide, chloride, or ascorbate. In a dry or soluble pharmaceutical composition SkQ1H 2  can be stabilized and protected from oxidation by a reducing agent such as, but not limited to, ascorbate.
 
     Yet another approach to improve stability is to place the MTA, in reduced or oxidized form, in a “softgel” formulation, which is a gelatin-based capsule with a liquid filling. Softgel formulations of MTAs provide good bioavailability as the softgel dissolves in aqueous-miscible, oily liquid carriers such as mono- and digycerides of capric/caprylic acid (Capmul MCM), Miglyol oil 8122 (medium chain triglycerides). When the softgel is released in the body, it gets emulsified and provides drug dispersion at a high surface area. 
     Mono- and digycerides of capric/caprylic acid (Capmul MCM), Miglyol oil 8122 (medium chain triglycerides) can be used. Such oily carriers as they become part of a self-emulsifying system. Other exemplary stabilizing components are vitamin E/polyethylene glycol succinate, sorbitan monooleate, labrasol, and combinations thereof. Additionally, based on its oxidation potential, tocopherol, butylayed hydroxytoluene, and/or butylated hydroxy anisole can be included in the composition as an antioxidant. 
     Another approach for increasing stabilization of MTAs in solution is to create a nanosuspension of MTA (&lt;1000 nm) stabilized with, e.g., vitamin E/polyethylene glycol succinate. Netzsch wet milling (http://www.netzsch-grinding.com) can be used to achieve this nanosuspension. 
     Additionally, ethanol solutions of reduced MTA (such as SkQ1H 2 ) can be mixed with the asorbic and acid dried to create resulting solid or powder that is stable for several months. 
     Stable formulations in the form of oral tablets can be prepared by hot melt extrusion. This melt granulation technique maintains the polymorphic stability of the drugs and significantly improve their oral bioavailability. It can be achieved by co-blending the MTAs with macrogols (e.g., polyethylene glycols 3350, 6000, polyvinyl pyrrolidone, hydroxy propyl cellulose and Vitamin E TPSG) through a hot melt extruder, and compressing the resulting granulation into tablets or encapsulating into hard gelatin capsules. 
     Representative stable liquid and solid oral SkQ1 formulations are shown below (Table 2): 
     
       
         
               
             
           
               
                 TABLE 2 
               
               
                   
               
             
             
               
                 Oxidized SkQ1 
               
               
                 Solutions: 
               
               
                 SkQ1 in 20% (wt %) glycerol, prepared with phosphate buffer 
               
               
                 SkQ1 in 50% (wt %) 1,2-propylene glycol with pyruvic acid 
               
               
                 SkQ1 in 50% (wt %) 1,2-propylene glycol with lactic acid 
               
               
                 Solid compositions: 
               
               
                 SkQ1 with PEG-4000 
               
               
                 SkQ1 with dextran 
               
               
                 SkQ1 with p-aminobenzoic acid (p-ABA) 
               
               
                 SkQ1 with dextran and p-ABA 
               
               
                 SkQ1 with myoinosite 
               
               
                 SkQ1 with pyruvic acid and Pearlitol 200 
               
               
                 SkQ1 with pyruvic acid and microcrystalline cellulose 
               
               
                 SkQ1 with pyruvic acid and F-Melt C 
               
               
                 SkQ1 with pyruvic acid and Syloid FP 
               
               
                 SkQ1 with citric (or tartaric acid, or lactic acid, or glycine) and 
               
               
                 Pearlitol 200 
               
               
                 SkQ1 with citric acid (or tartaric acid, or lactic acid, or glycine) and 
               
               
                 crocrystalline cellulose 
               
               
                 SkQ1 with citric acid (or tartaric acid, or lactic acid, or glycine) and 
               
               
                 F-Melt C 
               
               
                 SkQ1 with citric acid (or tartaric acid, or lactic acid, or glycine) and 
               
               
                 Syloid FP 
               
               
                 SkQ1H 2  (reduced form) 
               
               
                 Solutions: 
               
               
                 SkQ1H 2  (0.11M) with ascorbic acid (10 eq) in 55% EtOH 
               
               
                 SkQ1H 2  (7.4 mM) with ascorbic acid (5 eq) and sorbite (20 wt parts) in 
               
               
                 30% 1,2-propylene glycol 
               
               
                 Solid compositions: 
               
               
                 SkQ1H2 (1 eq) with ascorbic acid (&gt;2 molar eq) with PEG-4000 
               
               
                 SkQ1H2 (1 eq) with ascorbic acid (&gt;2 molar eq) with dextran 
               
               
                 SkQ1H2 (1 eq) with ascorbic acid (&gt;10 molar eq) with PEG-4000 
               
               
                 SkQ1H2 (1 eq) with ascorbic acid (&gt;10 molar eq) with dextran 
               
               
                 SkQ1H 2  (1 eq) with sorbite (30 wt parts) 
               
               
                 SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and sorbite (30 wt parts) 
               
               
                 SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and glucose (10 wt parts) 
               
               
                 SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and lactose monohydrate 
               
               
                 (10 wt parts) 
               
               
                 SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and Pearlitol 200 (30 wt parts) 
               
               
                 SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and microcrystalline cellulose 
               
               
                 (30 wt parts) 
               
               
                 SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and F-Melt C (30 wt parts) 
               
               
                 SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and Syloid FP (30 wt parts) 
               
               
                   
               
             
          
         
       
     
     SkQ1H 2  in the from of light powder was prepared to almost a 100% yield by the reduction of SkQ1 with ascorbic acid or any other suitable reducing agent in alcohol/water mixture followed by isolation by either extraction with chloroform or any other suitable solvent, or by precipitation from water followed by centrifugal separation, or by column (silica gel) chromatography or by method HPLC RP. The isolated material was characterized by 1H NMR, LC/MC and elemental analysis data. 
     The sample was proved to have excellent stability for 1 month at RT or several months at 4° C. in darkness under inert atmosphere without any humidity access (Table 17). The sample also can be stabilized by being dissolved in any deoxygenated anhydrous and aprotic solvents. The reduced form of SkQ1H 2  quickly oxides to the original form of SkQ1 when exposed to air or wet atmosphere or dissolved in water or any protonic solvent (Table 18). 
     The stability of SkQ1H 2  in solid compositions is strongly dependent on dryness of the composition as well as dryness of excipients and other components. Humidity of ambient atmosphere and presence of air also play a crucial role in oxidation of SkQ1H 2  into SkQ1 followed by degradation of the latter. 
     II. Treatments 
     In vivo and in vitro experiments demonstrate the ability of MTAs including, but not limited to, SkQ1 and SkQR1, to prevent and treat diabetes and disorders related to diabetes (Example 2). Such in vivo and in vitro experiments also demonstrate that liquid solutions of MTAs, including but not limited to SkQ1 and SkQR1, can be used for prevention and treatment of inflammatory diseases and related conditions such as septic shock and/or systemic. For example, these MTA-based liquid formulations with acceptable stability combined with results showing efficacy in models of diabetes, inflammation, septic shock, and related disorders (Examples 2-7). 
     SkQ1 treatment also prevented disassembling of intracellular contacts and cytoskeleton reorganization caused by TNFa (data obtained by microscopy studies of VE-cadherin, beta-cathenin and F-actin). Thus, SkQ1 was shown to be effective in protecting endothelial cells against the cytokine-caused dysfunction of endothelial barrier, and thus can be used for prevention and treatment of many pathological conditions including diabetes, atherosclerosis, aging, and chronicle inflammatory diseases. 
     Additionally, SkQ1 decreases the phosphorylation and degradation of IkBa caused by TNFα. NFκB is known to be permanently active in many inflammatory diseases, such as inflammatory bowel disease, arthritis, sepsis, gastritis, asthma and atherosclerosis (Monaco et al. (2004)  PNAS.,  101:5634-9). SkQ1 was shown to prevent activation of NFκB, a key inhibitor of NFκB activity associated with elevated mortality, especially from cardiovascular diseases (Venuraju et al. (2010)  J. Am. Coll. Cardiol.,  55:2049-61). In addition, SkQ1 was shown to prevent translocation of transcription factor p65 (RelA) from the cytoplasm to the nucleus, thereby potentially decreasing pathological consequences. 
     Reference will now be made to specific examples illustrating the invention. It is to be understood that the examples are provided to illustrate certain embodiments and that no limitation to the scope of the invention is intended thereby. 
     EXAMPLES 
     Example 1 
     Stable Formulations of Reduced Form of SkQ1 (SkQ1H 2 ) 
     SkQ1H 2 , a reduced quinole form of SkQ, was prepared as follows: 10 ml SkQ1H 2  solution (with concentration 1 mg/ml) in ethanol was thoroughly mixed with 200 mg ascorbic acid and then vacuum dryed. The resulting powder contained 95% ascorbic acid and 5% SkQ1H 2 , and demonstrated acceptable stability at several storage temperatures. For example, in the accelerated decay experiment, SkQ1 purity was reduced from initial 98.7% to 95.1% after storage for 12 d at 60° C. From these results it can be calculated that storage for 1 year at 4° C. will result in approximately 3.5% loss from the initial concentration of the active compound SkQ1 which has acceptable stability. 
     Alternatively, a dry mixture of SkQ1H 2  and ascorbic acid is prepared by dissolving 10 mg SkQ1H 2  in 10 ml ascorbic acid solution (20 mg/ml) and dried under vacuum. 
     Yet another way to prepare an SkQ1-ascorbic acid mixture is to mix 5 ml SkQ1H 2  solution in ethanol (2 mg/ml), with 5 ml ascorbic acid solution in water (40 mg/ml), and vacuum dry. The reduced form of SkQH 2  is stabilized in ascorbic acid solution, eliminating the drying stage, and thus the corresponding liquid formulation. 
     Example 2 
     Effect of Liquid MTA Formulations on Diabetes 
     A. Alloxan Animal Studies 
     Alloxan is a well-known diabetogenic agent widely used to induce type 2 diabetes in animals (Viana et al. (2004)  BMC Pharmacol.,  8:4-9). 
     Induction of the alloxan diabetes was performed as follows: Two groups of laboratory rats (20 animals in each group) with free food and water access fed a 250 nM solution of SkQ1 for 10 d. The daily rat consumption was 60 ml water solution (containing 15 nmoles SkQ1). The average weight of rats was 300 g. Thus, rats consumed approximately 50 nmol/kg body weight per day. Two other groups of animals did not receive SkQ1. After 10 d, rats were subcutaneously (in the area of the thigh) injected with alloxan dissolved in isotonic salt solution of 0.9% w/v of NaCl (100 mg/kg body weight; groups “Alloxan+SkQ1” and “Alloxan.” Control animals were injected with salt solution without alloxan (groups “Control+SkQ1” and “Control”). After injection, the rats continued to drink water containing SkQ1 (250 nM) during 14 d (group “Alloxan+SkQ1”) or were kept without SkQ1 (group “Alloxan”). 
     Data on glucose blood level was measured by the glucose oxidase method (Saifer et al. (1958)  J. Lab. Clin. Med.,  51:445-460) after 2 weeks of alloxan injection. The results are presented in  FIG. 1 . All data are presented as the mean+/−SE. 
     Animals consuming SkQ1 after alloxan injection had about 2-fold lower blood glucose compared to mice without SkQ1 treatment. 
     These results demonstrate that stabilized MTAs, e.g. SkQ1, are useful for the prevention and treatment of diabetes mellitus and its complications. 
     In another experiment, 200 g to 250 g Wistar male rats (age 7 to 8 weeks) were divided into 3 groups, 12 to 15 animals each and were injected with alloxan 125 mg/kg intraperitoneally (i.p.) after overnight fasting. Control animals were injected with saline (0.9% NaCl). The stabilized formulation (1% ethanol, 5 ml/kg) and SkQ1H 2  (5 eq ascorbic acid, 30 wt parts sorbite) in a dosage of 1250 nmol/kg was administered intragastrically (i.g.) by gavage once daily for 2 weeks before and 1 week after alloxan administration. Blood samples from tail vein were collected after overnight fasting and glucose levels were measured before alloxan administration and 1 d, 2 d, 3 d, and 7 d later by the conventional glucose-oxidase method. Seven days after alloxan administration rats were subjected to a glucose tolerance test. Rats were given glucose 3 g/kg i.g. Blood glucose levels were measured before glucose injection and 15 min, 30 min, 60 min, and 90 min later. 
     The following results were obtained (Table 3): 
                                                       TABLE 3                       Saline +       Alloxan + SkQ1H 2             vehicle   Alloxan + vehicle   formulation                                    Maximum glucose   6.7   17.6   13.9       conc. in blood, mM       Integrated glucose   500   1194   947       con. in blood (area       under curve,       a.u = mM × min)                    
B. Diabetic Mouse Studies
 
     Mice carrying mutation in leptin receptor gene (C57BLKS-Leprdb/J mice, or db/db mice) are known to be affected by glucose metabolic disorders. These mice are used as type II diabetes model with many of the characteristics of human disease including hyperphagia, hyperglycemia, insulin resistance, progressive obesity (Hummel et al. (1966)  Science,  153:1127-1128). 
     SkQ1 in 20% glycerol, as described below in Example 8 (250 nmol/kg per day) was orally administered to 10 to 12 week old homozygous db/db mice (n=8), while vehicle db/db (n=8) and non-diabetic control heterozygous db/++ (n=5) mice for 12 weeks. The hepatic TBA-reactive substance content (MDA) was determined by assay according to the method of Mihara et al. ((1978)  Anal. Biochem.,  86:271-278). 
     As shown in  FIG. 2 , elevated glucose levels induce oxidative stress reflected by the increased MDA levels in the liver of db/db mice. The increase of MDA level reflects stimulation of lipid peroxidation which in turn is considered responsible for the impairment of endothelial cells, capillary permeability, and fibroblast and collagen metabolism, major factors of pathologies associated with diabetes. The stabilized solution of SkQ1 significantly lowered MDA levels in the liver of diabetic db/db mice, thus indicating decreased rate of lipid peroxidation and decreased damage of the liver. 
     Example 3 
     Effect of Stabilized MTA on Wound Healing 
     Wound healing was studied in two series using 6 months old C57BLKS-Leprdb/J mice (db/db) homozygous and heterozygous C57BLKS-Leprdb/J mice (db/+) mice. These mice are used as type II diabetes model with impaired wound healing (Michaels, et al. (2007)  Wound Repair and Regeneration,  15:665-670). 
     The mice were daily administered 250 nmol/kg body weight per day with the pharmaceutical form of SkQ1 in 20% glycerol as described in Example 8) during period of time from 10 weeks to 12 weeks. Control groups of db/db and db/+ mice were not treated with SkQ1. Full-thickness dermal wounds were made under anesthesia of ketamine (80 mg/kg). Animals were kept in plastic cages under standard temperature, light, and feeding regimes. 7 days after wounding, animals were sacrificed by decapitation. The wounds were excised, fixed in 10% formalin in standard PBS buffer, histologically processed, and embedded in paraffin. Histological sections of central part of the wounds were cut and stained with hematoxylin and eosin. The sections were immunohistochemically stained for markers of endothelial cells (CD31), macrophages (f4/80), and myofibroblasts (smooth muscle α-actin). ImageJ software (National Institutes of Health (NIH) http:/rsb.info.nih.gov/ij/) was used to calculate total amount of cells, number of neutrophils, macrophages and vessel density (vessel area/granulation tissue area*100) on the microphotographs of wound sections. For each animal 100 mm 2  of section area was analyzed. Wound epithelization rate was assessed in % as ratio of epithelized wound area to total wound area on tissue section*100. For statistical analysis nonparametric Mann-Whitney U-test was used. Data are shown as means±S.E.M. 
     As shown in  FIGS. 3   a ,  3   b  and  3   c , the stabilized pharmaceutical form of SkQ1 is able to accelerate wound healing by decreasing neutrophil infiltration, increasing vascularization, and increasing the rate of epithelization in diabetic mice. 
     Example 4 
     Effect of Stabilized MTA on Inflammation and Septic Shock 
     Septic shock is known to activate numerous inflammatory pathways in an organism leading to death. The lipopolysaccharide (LPS)-induced septic shock mouse is widely accepted model in pharmacological and biological research (Villa et al. (2004)  Meth. Molec. Med.,  98:199-206). 
     Induction of the septic shock was performed as follows: 43 male BALB/c mice with free food and water access were divided onto 4 experimental groups. Group “K” got water without drugs. Groups “SkQ 50,” “SkQ 250,” and “SkQ 1250” were daily parenterally treated with pharmaceutical form of SkQ1 in water comprising 50 nmol/kg, 250 nmol/kg, and 1250 nmol/kg accordingly. After 3 weeks of SkQ1 treatment animals were intraperitoneally injected with 250 mg/kg LPS and 700 mg/kg D-galactosamine (D-GalN) inducing septic shock leading to death of 50% of untreated control animals (LD50 dose). Death of animals were registered after 4 d of septic shock induction. 
     The results of the experiment are shown on  FIG. 4 . The survival of mice following LPS/D-GalN treatment was significantly improved by SkQ1. The statistically significant effect was shown for a dose of 50 nmol/kg (p=0.03). 
     These results clearly indicate that SkQ1 acts as an anti-inflammatory agent having a therapeutic application for septic shock treatment. 
     In other studies, BALB/c mice with free food and water access are divided onto 4 experimental groups. Group “K” receive 20% glycerol without drugs. Groups “SkQ 50,” “SkQ 250,” and “SkQ 1250” are daily parenterally treated with pharmaceutical form of SkQ1 in 20% glycerol (Example 8) comprising 50 nmol/kg, 250 nmol/kg, and 1250 nmol/kg accordingly. After 3 weeks of SkQ1 treatment animals are intraperitoneally injected with 250 mg/kg LPS and 700 mg/kg D-galactosamine (D-GalN) inducing septic shock leading to death of 50% of untreated control animals (LD50 dose). Death of the animals is registered after 4 d of septic shock induction. 
     Example 5 
     Effect of Stabilized MTA on Arthritis 
     The collagen-induced arthritis (CIA) rat model was used to examine the susceptibility of rheumatoid arthritis (RA) to treatment with potential anti-arthritic agents (Griffiths et al. (2001)  Immunol. Rev.,  184:172-83). 
     Thirty Wistar rats with free food and water access were injected with complete Freund adjuvant and 250 mg type II collagen to induce CIA. Starting from 14 d and from 24 d after injection, two groups of 10 animals in each were daily fed with pharmaceutical form of SkQ1 in water comprising 250 nmol/kg body weight per day (groups “SkQ1 from day 14” and “SkQ1 from day 24”; Group “Control” received water without drugs). 
     As shown in  FIG. 5 , SkQ1 reduced the number of animals with apparent inflammation, i.e. animals with increased paw volumes measured by water manometry compared to control group. Hence, SkQ1 possesses anti-inflammatory and anti-arthritic effects. 
     In other studies, Wistar rats with free food and water access are injected with complete Freund adjuvant and 250 mg type II collagen to induce CIA. Starting from 14 d and from 24 d after injection, two groups of animals in each are daily fed with pharmaceutical form of SkQ1 in 20% glycerol (Example 8) comprising 250 nmol/kg body weight per day (groups “SkQ1 from day 14” and “SkQ1 from day 24”; Group “Control” received water without drugs). 
     Example 6 
     Effect of Stabilized MTA on Inflammation Associated with Coronary Heart Disease 
     Intense cytokine production induced by inflammation may lead to death of endothelial cells which, along with increased oxidative stress and vascular inflammation, leads to endothelial dysfunction and increases the risk for coronary artery disease. 
     Human endothelial cell line EA.hy926 (ATCC Collection; catalog number CRL-2922) was used as a model of vascular endothelium. This cell line is similar to primary HUVEC cell line (Edgell et al. (1983)  PNAS,  80(12):3734-7; Edgell et al. (1990)  In Vitro Cell Dev Biol.,  26(12):1167-72) and widely used as a relevant model for inflammation studies (Riesbeck et al. (1998)  Clin. Vaccine Immunol.,  5:5675-682). 
     Accordingly, human endothelial cells EA.hy926 were pre-incubated with 0.2 nM SkQR1 or 2 nM SkQ1 solution in Dulbecco&#39;s Modified Eagle&#39;s Medium (DMEM) supplemented with 10% of fetal serum (Example 1) for 4 d. After that the cells were incubated overnight with fresh DMEM medium with 0.2% of fetal serum. The cells were incubated 2 d with TNF-α (0.25 ng/ml to 50 ng/ml) and cell death was monitored using standard MTT test (Berridge et al. (1996)  Biochemica,  4:14-9). The data from this assay is shown as means±S.E. at least for 3 separate experiments. 
     As shown in  FIG. 6 , both SkQ1 and SkQR1 greatly reduced cell death compared to control without MTA. Thus, SkQ1 and SkQR1 were shown to be effective substance protecting endothelial cells against cytokine&#39;s inflammatory action and can be used for prevention and treatment of coronary heart disease including atherothrombosis. 
     Example 7 
     Effect of Stabilized MTA on Vascular Dysfunction 
     A. In Vitro Studies 
     Inflammatory cytokines induce expression of ICAM-1 (Inter-Cellular Adhesion Molecule 1). ICAM-1 is a key molecule functioning in intercellular adhesion process and transmigration of leukocytes across vascular endothelia during inflammatory response. Expression of ICAM-1, as well as inflammatory cytokines including IL-6 and IL-8, is elevated under many pathological conditions including diabetes, atherosclerosis, aging, and chronicle inflammatory diseases. 
     The effects of SkQ1 on ICAM-1 mRNA expression and cytokines (IL-6, IL-8) protein secretion induced by TNF-α in EAhy926 human endothelial cells (ATCC collection; catalog number CRL-2922) were examined. TNF-α is a central proinflammatory cytokine stimulating expression of cell adhesion molecules and many inflammatory cytokines. Anti-inflammatory properties of many drugs often rely on their ability to inhibit expression of pro-inflammatory cytokines induced by TNF-α using EAhy926 endothelial cells (Edgell et al. (1983)  Proc. Natl. Acad. Sci. USA,  80:3734-7; Lombardi et al. (2009)  Eur. J. Cell. Biol.,  88:731-42; Manea et al. (2010)  Cell Tissue Res.,  340:71-9). 
     300,000 cells were plated on 60 mm 2  culture dishes and after attachment were treated with an SkQ1 solution (0.2 nM in DMEM medium with 10% fetal serum) for 4 d, and then stimulated with TNF-α (0.05 ng/ml for 4 h for ICAM-1 or 5 ng/ml for 15 h for cytokines, respectively). ICAM-1 mRNA expression was determined by real-time PCR (Okada et al. (2005)  Invest. Ophtalmol. Vis. Sci.,  46:4512-8). Secretion of IL-6 and IL-8 was evaluated by ELISA (Toma et al. (2009)  Biochem. Biophys. Res. Commun.,  390:877-82; Volanti et al. (2002)  Photochem. Photobiol.,  75:36-45.) The data is shown as means±S.E. at least for 3 separate experiments. 
     The results shown in  FIG. 7   a  confirm SkQ1 to be and effective vascular anti-inflammatory substance that prevents excessive expression of inflammatory cytokines and ICAM-1. Thus, MTAs are useful for prevention and treatment of vascular pathologies including atherosclerosis. 
     B. In Vivo Studies 
     As described above in Example 7A, above, the expression of ICAM-1 is elevated under many pathological vascular conditions. SkQ1 efficacy in reducing ICAM-1 expression in vivo was tested on mice. 30 hybrid male C57Black/CBA mice were divided into 3 experimental groups (10 animals in each group) at the beginning of the experiment. The group “Young mice” included mice at the age of 6 months. Groups “Old mice” and “Old mice, SkQ1” included mice at the age of 24 months. The group “Old mice, SkQ1” had free access to drinking water with 100 nM water-dissolved SkQ1 per 1 kg of body weight for 7 months. After this period, the animals were decapitated. Aortas were excised, and total RNA was isolated using DNeasy Blood and Tissue kit (QIAGEN), reverse-transcribed into cDNA, and used for quantitative real-time PCR analysis of ICAM-1 mRNA level. For the normalization procedure the average geometry of expression levels of housekeeping genes GAPDH and RPL32 was used Data are shown as means±S.E.M. 
     As shown on  FIG. 7   b , SkQ1 significantly lowered ICAM-1 mRNA levels in treated old mice compared to the control group and approaches the level of ICAM-1 in young mice. 
     The results demonstrate that SkQ1 prevents the age-related increase of ICAM-1 expression in the vascular endothelium. Thus, SkQ1 can be used for prevention of age-related vascular pathologies including atherosclerosis. 
     In other studies, hybrid male C57Black/CBA mice are divided into 3 experimental groups, “young,” “old,” and “old mice, SkQ1,” as described above. The third group receives SkQ1 in 20% glycerol comprising 250 nmol/kg body weight per day dose up to 7 months. The “old” group is the control and receives glycerol without drugs. After this period, the animals are decapitated. Aortas are excised, and total RNA is isolated using DNeasy Blood and Tissue kit (QIAGEN), reverse-transcribed into cDNA, and used for quantitative real-time PCR analysis of ICAM-1 mRNA level. For the normalization procedure the average geometry of expression levels of housekeeping genes GAPDH and RPL32 are used. Data are calculated as means±S.E.M. 
     Example 8 
     Preparation and Stability of Oxidized SkQ1 Formulations 
     1. SkQ1 in 20% (wt %) Glycerol and Phosphate Buffer 
     Glycerol (20 g) was diluted with phosphate buffer (80 g, 0.01 M KH 2 PO 4 , pH 4.77). A sample of SkQ1 (20 mg) was placed in a dark glass vial and dissolved in propylene glycol (0.2 mL) and diluted with an aliquot (19.8 ml) of the above solvent to 1 mM. 
     The stability of SkQ1 in the prepared solution was investigated by storage at RT and at 60° C. (Table 4). 
                                           TABLE 4                   SkQ1, %/degradation   SkQ1, %/degradation       Time, days   products, % (stored at RT)   products, % (stored at 60° C.)                                0   99.34/0     99.34/0         11   99.71/0     —       13   99.76/0     —       14   99.68/0     —       17   99.62/0     —       19   99.63/0.07   95.30/4.7       21   99.52/0.20   —       24   99.57/0.08   —       61   99.49/0.51   —                    
2. SkQ1 in 50% (wt %) 1,2-Propylene Glycol with Pyruvic Acid (10 Equivalents (eq) Relative to SkQ1)
 
     SkQ1 (50 mg) and pyruvic acid (71 mg, 10 eq) were placed in a dark glass vial and dissolved in 50% propylene glycol-water mixture (100 ml) to yield a 0.081 mM SkQ1 solution. 
     The stability of SkQ1 in the prepared solution was investigated by storage at 60° C. (Table 5). 
     3. SkQ1 in 50% (wt %) 1,2-Propylene Glycol with Lactic Acid (10 eq Relative to SkQ1) 
     SkQ1 (50 mg) and L(+)-lactic acid (73 mg, 10 eq) were placed in a dark glass vial and dissolved in 50% propylene glycol-water mixture (100 ml) to yield a 0.081 mM SkQ1 solution. 
     The stability of SkQ1 in the prepared solution was investigated by storage at 60° C. (Table 5). 
                                           TABLE 5               Time, days   SkQ1, %   SkQ1, %                                0   &gt;99.9   &gt;99.9       72   93.2   96.6                    
4. SkQ1 with PEG-4000
 
     A solution of 8 mg SkQ1 in 0.5 ml EtOH was mixed with 200 mg PEG-4000, and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at 4° C. in darkness (Table 6). 
                                           TABLE 6               Time, days   SkQ1, %   Degradation products, %                                18   &gt;99.9   &lt;0.01       19   99.83   0.17       20   99.80   0.20                    
5. SkQ1 with Dextran
 
     A solution of 10 mg SkQ1 in 0.75 ml EtOH was added to a solution of 100 mg dextran in 1 ml water. The mixture was vigorously stirred and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at 60° C. in darkness (Table 7). 
                                           TABLE 7               Time, days   SkQ1, %   Degradation products, %                                0   96.71   3.29       6   20.66   79.34       15   24.14   75.86       25   18.93   81.07                    
6. SkQ1 with p-Aminobenzoic Acid (p-ABA)
 
     A solution of 8 mg SkQ1 in 0.5 ml EtOH was added to a solution of 200 mg p-aminobenzoic acid (p-ABA) in 1.5 ml EtOH. The solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at RT in darkness (Table 8). 
                                           TABLE 8               Time, days   SkQ1, %   Degradation products, %                                0   100   0       30   58.42   41.58                    
7. SkQ1 with Dextran and p-ABA
 
     A solution of 10 mg SkQ1 in 0.75 ml EtOH was added to a solution of p-ABA (2 mg in 0.5 ml EtOH) and dextran (100 mg in 1 ml water). The mixture was vigorously stirred and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at 60° C. in darkness (Table 9). 
                                           TABLE 9               Time, days   SkQ1, %   Degradation products, %                                0   97.13   2.87       6   39.22   60.78       15   7.07   92.93                    
8. SkQ1 (1 eq) with Myoinosite (30 wt Parts Relative to SkQ1)
 
     45 mg myoinosite was added to a solution of 5 mg SkQ1 in 5 ml EtOH. The mixture was vigorously stirred and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at RT in darkness (Table 10). 
                                           TABLE 10               Time, days   SkQ1, %   Degradation products, %                                0   95.88   4.12       5   96.86   3.14       6   95.99   4.01       15   92.26   7.74                    
9. SkQ1 (1 eq) with Pyruvic Acid (10 eq) and Pearlitol 200 (30 wt Parts Relative to SkQ1)
 
     375 mg Pearlitol 200 was added to a solution of 12.5 mg SkQ1 and 17.8 mg (10 eq) pyruvic acid in 0.75 ml EtOH. The mixture was vigorously stirred and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at 60° C. in darkness (Table 11). 
     10. SkQ1 (1 eq) with Pyruvic Acid (10 eq) and Microcrystalline Cellulose (30 wt Parts Relative to SkQ1 
     375 mg microcrystalline cellulose was added to a solution of 12.5 mg SkQ1 and 17.8 mg (10 eq) pyruvic acid in 0.75 ml EtOH. The mixture was vigorously stirred and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at 60° C. in darkness (Table 11). 
     11. SkQ1 (1 eq) with Pyruvic Acid (10 eq) and F-Melt C (wt Parts Relative to SkQ1) 
     375 mg F-Melt C was added to a solution of 12.5 mg SkQ1 and 17.8 mg (10 eq) pyruvic acid in 0.75 ml EtOH. The mixture was vigorously stirred and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at 60° C. in darkness (Table 11). 
     12. SkQ1 (1 eq) with Pyruvic Acid (0 eq) and Syloid FP (30 wt Parts Relative to SkQ1) 
     375 mg Syloid FP was added to a solution of 12.5 mg SkQ1 and 17.8 mg (10 eq) pyruvic acid in 0.75 ml EtOH. The mixture was vigorously stirred and the solvent was evaporated to dryness. 
     The stability of SkQ1 in the prepared composition was investigated by storage at 60° C. in darkness (Table 11). 
     
       
         
               
               
             
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 11 
               
             
             
               
                   
               
               
                 Time, 
                 SkQ1, %/SkQ1H 2 , %, degradation products, % 
               
             
          
           
               
                 days 
                 (Sample 9) 
                 (Sample 10) 
                 (Sample 11) 
                 (Sample 12) 
               
               
                   
               
             
          
           
               
                 0 
                 &gt;99.9/&lt;0.05/ 
                 &gt;99.9/&lt;0.05/ 
                 &gt;99.9/&lt;0.05/&lt;0.05 
                 &gt;99.9/ 
               
               
                   
                 &lt;0.05 
                 &lt;0.05 
                   
                 &lt;0.05/ 
               
               
                   
                   
                   
                   
                 &lt;0.05 
               
               
                 14 
                 60.3/11.3/ 
                 50.2/25.8/24.0 
                 38.2/47.7/14.1 
                 57.9/1.4/ 
               
               
                   
                 28.4 
                   
                   
                 40.7 
               
               
                   
               
             
          
         
       
     
     The following SkQ1 preparations can also be formulated as described supra in Example 8:
         SkQ1 (1 eq) with citric (or tartaric acid, or lactic acid, or glycine, 10 eq) and Pearlitol 200 (30 wt parts in relation to SkQ1H 2 )   SkQ1 (1 eq) with citric acid (or tartaric acid, or lactic acid, or glycine, 10 eq) and microcrystalline cellulose (30 wt parts in relation to SkQ1H 2 )   SkQ1 (1 eq) with citric acid (or tartaric acid, or lactic acid, or glycine, 10 eq) and F-Melt C (30 wt parts in relation to SkQ1H 2 )   SkQ1 (1 eq) with citric acid (or tartaric acid, or lactic acid, or glycine, 10 eq) and Syloid FP (30 wt parts in relation to SkQ1H 2 )       

     Example 9 
     Preparation and Stability of Reduced SkQH 2  Formulations 
     13. SkQ1H2 (1 eq) Prepared in Situ by Reduction of SkQ1 and Ascorbic Acid (2 Molar eq) and PEG-4000 (10 wt Parts Relative to SkQ1H 2 ) 
     A solution of 10 mg SkQ1 in 0.6 ml EtOH was added to solution of 5.7 mg (2 eq) ascorbic acid in 0.1 ml water. The mixture was stirred until reduction to SkQ1H 2  completed (about 1 h). Then 100 mg PEG-4000 was added. The mixture was vigorously stirred for 30 min and the solvent evaporated to dryness. 
     The stability of SkQ1H 2  in the prepared composition was investigated by storage at 4° C. in darkness (Table 12). 
     14. SkQ1H2 (1 eq Prepared in Situ by Reduction of SkQ1 with Ascorbic Acid (2 Molar eq) and Dextran) 
     A solution of 10 mg SkQ1 in 0.6 ml EtOH was added to solution of 5.7 mg (2 eq) ascorbic acid in 0.1 ml water. The mixture was stirred until reduction to SkQ1H 2  completed (about 1 h). Then a solution of 100 mg dextran in 1 ml water was added. The mixture was vigorously stirred for 30 min and the solvent was evaporated to dryness. 
     The stability of SkQ1H 2  in the prepared composition was investigated by storage at 4° C. in darkness (Table 12). 
                                                                                         TABLE 12                       (Sample 13)   (Sample 14)                        Degra-           Degra-                   dation           dation       Time,   SkQ1,   SkQ1H 2 ,   products,   SkQ1,   SkQ1H 2 ,   products,       days   %   %   %   %   %   %                    0   14.65   85.35   &lt;0.05   3.61   96.39   &lt;0.05       1   7.72   92.28       2.80   97.20           4   59.12   40.88       98.57   1.43           6   57.53   42.47       99.55   0.45           7   54.16   45.84       99.26   0.74           10   54.22   45.78       98.93   1.07                    
15. SkQ1H2 (1 eq) Prepared in Situ by Reduction of SkQ1 with Ascorbic Acid (10 Molar eq) and Dextran (10 wt Parts Relative to SkQ1H 2 )
 
     A solution of 10 mg SkQ1 in 0.6 ml EtOH was added to solution of 28.5 mg (10 eq) ascorbic acid in 0.25 ml water. The mixture was stirred until reduction to SkQ1H 2  was completed (about 30 min). A solution of 100 mg dextran in 1 ml water was then added. The mixture was vigorously stirred for 30 min and the solvent evaporated to dryness. 
     The stability of SkQ1H 2  in the prepared composition was investigated by storage at 60° C. in darkness (Table 13). 
     16. SkQ1H2 (1 eq) (Prepared in Situ by Reduction of SkQ1 with Ascorbic Acid (&gt;10 Molar eq) with Dextran and p-ABA (10 wt Parts Relative to SkQ1H 2    
     A solution of 10 mg SkQ1 in 0.6 ml EtOH was added to solution of 28.5 mg (10 eq) ascorbic acid in 0.25 ml water. The mixture was stirred until reduction to SkQ1H 2  was completed (about 30 min). A solution of 100 mg dextran in 1 ml water and a solution of 2 mg p-ABA in 0.5 ml EtOH were then added. The mixture was vigorously stirred for 30 min and the solvent evaporated to dryness. 
     The stability of SkQ1H 2  in the prepared composition was investigated by storage at 60° C. in darkness (Table 13). 
                                                                                         TABLE 13                       (Sample 15)   (Sample 16)                        Degra-           Degra-                   dation           dation       Time,   SkQ1,   SkQ1H 2 ,   products,   SkQ1,   SkQ1H 2 ,   products,       days   %   %   %   %   %   %                    0   2.35   92.59   5.06   0.74   98.65   0.61       6   4.26   91.66   4.08   2.72   97.16   0.12       15   5.11   94.27   0.62   8.49   91.12   0.39       25   5.71   88.69   5.6   11.07   86.62   2.31                    
17. SkQ1H 2  Powder
 
     A solution of 2 g SkQ1 in 40 ml EtOH was added to a solution of 5.7 g ascorbic acid in 60 ml water. The mixture was stirred until reduction to SkQ1H 2  was completed (about 30 min). Completion of reduction can be detected as the solution becomes colorless. The solvent was then evaporated off and the residue was partitioned between water (50 ml) and CHCl 3  (150 ml). The organic layer was washed with water (2×25 ml), dried with anhydrous sodium sulfate, filtered, and evaporated. 
     The yield of SkQ1H 2  was 2 g (approx 100% yield) in the form of light powder. The stability results are shown below (Table 14 and Table 15). 
     
       
         
               
               
               
             
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 14 
               
             
             
               
                   
               
               
                   
                 Storage at RT 
                 Storage at 60° C. 
               
             
          
           
               
                   
                   
                   
                 Degra- 
                   
                   
                 Degra- 
               
               
                   
                   
                   
                 dation 
                   
                   
                 dation 
               
               
                 Time, 
                 SkQ1H 2 , 
                 SkQ1, 
                 products, 
                 SkQ1H 2 , 
                 SkQ1, 
                 products, 
               
               
                 days 
                 % 
                 % 
                 % 
                 % 
                 % 
                 % 
               
               
                   
               
             
          
           
               
                 0 
                 98.99 
                 1.01 
                 &lt;0.1 
                 99.2 
                 0.75 
                 0.05 
               
               
                 3 
                 99.34 
                 0.66 
                 &lt;0.1 
                 — 
                 — 
                 — 
               
               
                 5 
                 99.37 
                 0.63 
                 &lt;0.1 
                 99.45 
                 0.55 
                 0 
               
               
                 7 
                 99.14 
                 0.71 
                 &lt;0.1 
                 100 
                 0 
                 0 
               
               
                 11 
                 99.12 
                 0.83 
                 &lt;0.1 
                 99.76 
                 0.19 
                 0.05 
               
               
                 17 
                 99.49 
                 0.28 
                 &lt;0.3 
                 98.61 
                 1.24 
                 0.15 
               
               
                 28 
                 99.45 
                 0.50 
                 &lt;0.1 
                 88.7 
                 11.04 
                 0.26 
               
               
                   
               
             
          
         
       
     
                                                                                         TABLE 15                       55% EtOH in water   CH 2 Cl 2                          Degra-           Degra-                   dation           dation       Time,   SkQ1H 2 ,   SkQ1,   prod-   SkQ1H 2 ,   SkQ1,   prod-       h   %   %   ucts, %   %   %   ucts, %                    0   97.79   2.21   0   97.79   2.21   0       0.5   90.79   9.21   0   95.99   4.01   0       1.48   85.24   14.76   0   94.32   5.68   0       2.8   67.43   32.57   0   93.58   6.42   0       3.44   52.17   47.83   0   94.43   5.57   0       4.37   43.43   56.57   0   92.82   7.18   0       23.23   16.55   82.39   1.06   89.61   9.30   0.97       143.45   9.63   77.23   13.14   82.11   16.53   1.36       (~6                               days)                    
18. SkQ1H 2  (1 eq) with Sorbite (30 wt Parts Relative to SkQ1H 2 )
 
     A solution of 20 mg SkQ1H 2  in 1.3 ml EtOH was added to a solution of 600 mg sorbite in 1.3 ml water. The solvent was evaporated to dryness. The residue was additionally dried with diphosphorous pentoxide (P 2 O 5 ) under reduced pressure. 
     The stability of SkQ1H 2  in the prepared composition was investigated by storage at 60° C. in darkness (Table 16). 
                                                   TABLE 16               Time, days                   (at 60° C.)   SkQ1H 2 , %   SkQ1, %   Degradation products, %                                0   99.01   0.61   0.38       4   90.8   8.7   0.5       7   90.2   9.4   0.4       11   88.8   10.7   0.5       15   89.1   10.4   0.5       28   42.9   5.3   51.8                    
19. SkQ1H 2  (1 eq) with Ascorbic Acid (0-5 eq) and Sorbite (30 wt Parts Relative to SkQ1H 2 )
 
     Method 1: 
     A solution of 20 mg SkQ1H 2  in 1.3 ml EtOH was added to a solution of 28.4 mg (5 eq) ascorbic acid and 600 mg sorbite in 1.3 ml water. The solvent was evaporated to dryness. The residue was additionally dried with P 2 O 5  under reduced pressure. 
     Method 2: 
     20 mg SkQ1H 2  and 28.4 mg (5 eq) ascorbic acid were added to sorbite (600 mg) melted in a glass vial (bath temperature 110° C.) slowly under vigorous stirring and stirring continued for 1 hr. The mixture was cooled to RT and vigorously triturated to provide a microcrystalline powder. 
     The stability of SkQ1H 2  in the compositions prepared by both methods was investigated by storage at 60° C. and 4° C. in darkness (Table 17). 
     
       
         
               
               
               
             
               
               
               
               
               
             
           
               
                   
                 TABLE 17 
               
             
             
               
                   
                   
               
               
                   
                 Degradation products, 
                   
               
               
                   
                 (total, %/number of impurities 
               
               
                   
                 with content &gt;0.5%) 
               
             
          
           
               
                   
                 SkQH 2 , % 
                 SkQ1, % 
                 20 days at 60° C. 
                 1 year at 4° C. 
               
               
                   
                   
               
               
                   
                 97.753 
                 1.209 
                 1/0 
                 0.3/0 
               
               
                   
                   
               
             
          
         
       
     
     The following SkQ1H 2  preparations in ascorbic acid are also prepared as in Example 19 supra:
         SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) with magnesium stearate (10 wt % in relation to SkQ1H 2 ) and glucose (10 wt parts in relation to SkQ1H 2 )   SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) with magnesium stearate (10 wt % in relation to SkQ1H 2 ) and lactose monohydrate (10 wt parts in relation to SkQ1H 2 )   SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and Pearlitol 200 (30 wt parts in relation to SkQ1H 2 )   SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and microcrystalline cellulose (30 wt parts in relation to SkQ1H 2 )   SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and F-Melt C (30 wt parts in relation to SkQ1H 2 )   SkQ1H 2  (1 eq) with ascorbic acid (0-5 eq) and Syloid FP (30 wt parts in relation to SkQ1H 2 )
 
20-22 and 26-30. SkQ1H 2  with Ascorbic Acid (0-5 eq) and Glucose
       

     Method 3: 
     A solution of 20 mg SkQ1H 2  in 1.3 ml EtOH was added to 2 mg magnesium stearate and solution of ascorbic acid (quantities as listed in the Table 18) and 600 mg glycose in 1.3 ml water (1.3 mL). The solvent was evaporated to dryness. The residue was additionally dried with P 2 O 5  under reduced pressure. 
     Method 4: 
     20 mg SkQ1H 2 , 2 mg magnesium stearate, ascorbic acid (quantities as listed in Table 18) and 600 mg anhydrous glycose were mixed and vigorously triturated. 
     The stability of SkQ1H 2  in compositions prepared by Methods 3 and 4 was investigated by storage at 60° C. in darkness (Table 18). 
     23.-25. SkQ1H 2  with Ascorbic Acid (0-5 eq) and Lactose Monohydrate 
     The compositions were prepared as described above in Method 3 or 4 using lactose monohydrate instead of glycose. 
     The stability of SkQ1H 2  in compositions prepared by both methods was investigated by storage at 60° C. in darkness (Table 18). 
                                                                                                     TABLE 18                       Formulation       Degradation products, total,           (stabilizers and excipients, amounts       %/number of impurities           are given in relation to SkQ1H 2         with content &gt;0.5%            Sample   Asc. acid,       L (+)-   Mg   Method of   20 d at   1 year at       No   eq   Glycose   Lactose × H 2 O   Stearate   preparation   60° C.   4° C.                    22   1   ~10 wt parts   —   10 wt %   4   &gt;30/7   ~6/2       23   3   ~10 wt parts   —   10 wt %   4   &gt;12/9    &lt;3/1       24   0.3   ~10 wt parts   —   10 wt %   4    &gt;9/7    &lt;3/1       25   1   —   ~10 wt parts   10 wt %   4   &gt;12/7   4.6/1       26   3   —   ~10 wt parts   10 wt %   4    &gt;9/6    &lt;3/2       27   0.3   —   ~10 wt parts   10 wt %   4   &gt;10/5   3.9/2       28   1   ~10 wt parts   —   10 wt %   3       ~6/3   2.8/0       29   2   ~10 wt parts   —   10 wt %   3    4.4/1   2.6/0       30   3   ~10 wt parts   —   10 wt %   3    4.2/0       2/0       31   5   ~10 wt parts   —   10 wt %   3    3.6/0   1.6/0       32   0.3   ~10 wt parts   —   10 wt %   3   3.5/3   —                               (7 d at                                   60° C.)                    
31. SkQ1H 2  with Ascorbic Acid in 55% EtOH
 
     A solution of pure SkQ1H 2  (1 g in 5 ml EtOH) was added to solution of ascorbic acid (2.85 g (10 eq) in 10 ml water). 
     The stability of SkQ1H 2  in the prepared solution was investigated by storage at RT in darkness (Table 19). 
                                                   TABLE 19               Time, h   SkQ1H 2 , %   SkQ1, %   Degradation products, %                                0     99.73   0.27   &lt;0.01       1.5   99.07   0.93   &lt;0.01        68 (~3 days)   99.05   0.59   &lt;0.4       118 (~5 days)   99.69   0.31   &lt;0.01       165 (~7 days)   99.74   0.26   &lt;0.01                    
32. SkQ1H 2  with Ascorbic Acid and Sorbite in 30% 1,2-Propylene Glycol
 
     A solution of pure SkQ1H 2  (50 mg in 1 ml 1,2-propylene glycol) was added to solution of ascorbic acid (67.4 mg (5 eq)) and sorbite (1.5 g) in 10 ml water. 
     The stability of SkQ1H 2  in the prepared solution was investigated by storage at 60° C. in darkness (Table 20). 
     
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                 TABLE 20 
               
               
                   
               
               
                 Time, days 
                 SkQ1, % 
                 SkQH 2 , % 
                 Degradation products, % 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 0 
                 0.18 
                 99.82 
                 0.00 
               
               
                 3 
                 1.03 
                 98.67 
                 0.30 
               
               
                 14 
                 28.34 
                 69.51 
                 2.15 
               
               
                 27 
                 51.9 
                 3.2 
                 44.9 
               
               
                   
               
             
          
         
       
     
     EQUIVALENTS 
     Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments described specifically herein. Such equivalents are intended to be encompassed in the scope of the following claims.