Patent Publication Number: US-2022226313-A1

Title: Combination therapies that include an agent that promotes glucose oxidation and an inhibitor of pyruvate dehydrogenase kinase

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims the benefit of, and priority to, U.S. Provisional Patent Application No. 62/856,369, filed Jun. 3, 2019, the contents of which are incorporated by reference. 
    
    
     FIELD OF THE INVENTION 
     The invention relates to methods and compositions for treating conditions, such as diabetes, cancer, and cardiovascular conditions. 
     BACKGROUND 
     Millions of people suffer from conditions associated with defective mitochondrial metabolism. For example, abnormalities in mitochondrial metabolism lead to decreased cardiac efficiency in many forms of heart disease, are a causative factor in maternally inherited diabetes, and contribute to metastasis of certain types of cancer. Mitochondria produce the majority of high-energy molecules within a cell, and defects in mitochondrial metabolism result in reduced energy production. In some clinical manifestations, such as heart failure, ischemic heart disease, and diabetic cardiomyopathies, the reduction is attributable to the reliance of affected tissues on fatty acids rather glucose as a source of energy. 
     SUMMARY 
     Aspects of the invention recognize that glucose oxidation requires less oxygen than does fatty acid oxidation, so tissues that use the latter are more susceptible to damage when blood supply is diminished. Therefore, therapies that promote glucose oxidation are needed to treat or prevent a wide variety of diseases and conditions. Accordingly, an agent that shifts cellular metabolism from fatty acid oxidation to glucose oxidation, such as trimetazidine or an analog, derivative, or prodrug thereof, is beneficial for treating mitochondrial disorders. 
     The invention additionally recognizes that glucose catabolism includes sequential anaerobic (i.e., oxygen-independent) and aerobic (oxygen-requiring) processes. Consequently, the invention further recognizes that a single agent may not result in complete breakdown of glucose given the different aspects of glucose catabolism. It has been discovered that a combination therapy in which both anaerobic (i.e., oxygen-independent) and aerobic (oxygen-requiring) processes of glucose catabolism are addressed provides a highly efficient treatment for mitochondrial disorders. In that manner, the invention provides combination therapies, methods, and compositions, that direct cellular metabolism by providing a first agent that triggers cells to break down glucose rather than fatty acids and a second agent that drives the aerobic steps of glucose catabolism to achieve complete oxidation of glucose. Thus, the invention provides therapies that optimize energy production in a variety of pathological conditions. 
     In certain aspects, the invention provides combination therapies that include a compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and a pyruvate dehydrogenase kinase (PDK) inhibitor. Together, these agents address both the anaerobic and aerobic processes of glucose catabolism, thereby providing a new approach for treating mitochondrial disorders. 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be trimetazidine, etomoxir, perhexiline, a PPAR agonist, a malonyl CoA decarboxylase inhibitor, dichloroacetate, or an analog, derivative, or prodrug of any of the aforementioned agents. 
     The compound that shifts cellular metabolism from fatty acid oxidation may be represented by formula (IV): 
     
       
         
         
             
             
         
       
     
     in which: 
     R 1 , R 2 , and R 3  are independently selected from the group consisting of H and a (C 1 -C 4 )alkyl group; 
     R 4  and R 5  together are ═O, —O(CH 2 ) m O—, or —(CH 2 ) m —, wherein m=2-4, or R 4  is H and R 5  is OR 14 , SR 14 , or (CH 2 CH 2 O) n H, wherein R 14  is H or a (C 1 -C 4 )alkyl group and n=1-15; and 
     R 6  is a single or multi-ring structure optionally substituted at one or more ring positions by a heteroatom, wherein each ring position optionally comprises one or more substituents. 
     One or more ring position of R 6  may be or include a substituent that includes a compound that promotes mitochondrial respiration, such as succinate, fumarate, malate, oxaloacetate, citrate, isocitrate, α-ketoglutarate, pyruvate, acetone, acetoacetic acid, β-hydroxybutpic acid, β-ketopentanoate, or β-hydroxypentanoate. The substituent may be or include a linker, such as (CH 2 CH 2 O) x , in which x=1-15. The substituent may be or include a NAD +  precursor molecule, such as nicotinic acid, nicotinamide, and nicotinamide riboside. 
     The substituent on a ring position of R 6  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     The substituent on a ring position of R 6  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     R 6  may be 
     
       
         
         
             
             
         
       
     
     The compound of formula (IV) may have a structure represented by one of formulas (IX) and (X): 
     
       
         
         
             
             
         
       
     
     The compound that shifts cellular metabolism from fatty acid oxidation may be represented by formula (V): 
     
       
         
         
             
             
         
       
     
     in which R 1 , R 2 , and R 3  are independently H or a (C 1 -C 4 )alkyl group; R 4  and R 8  together are ═O, —O(CH 2 ) m O—, or —(CH 2 ) m —, in which m=2-4, or R 4  is H and R 8  is H, OR 14 , SR 14 , or (CH 2 CH 2 O) n H, in which R 14  is H or a (C 1 -C 4 )alkyl group and n=1-15; R 9 , R 10 , R 12 , and R 13  are independently H or (CH 2 CH 2 O) z H, in which z=1-6; and R 11  comprises a compound that promotes mitochondrial respiration. 
     The compound that promotes mitochondrial respiration may be an intermediate of the citric acid cycle or a molecule that can be metabolized to enter the citric acid cycle. For example, the compound may be succinate, fumarate, malate, oxaloacetate, citrate, isocitrate, α-ketoglutarate, pyruvate, acetone, acetoacetic acid, β-hydroxybutpic acid, β-ketopentanoate, or β-hydroxypentanoate. 
     R 11  may include a linker, such as polyethylene glycol. For example, R 11  may include (CH 2 CH 2 O) x , in which x=1-15. 
     R 11  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     R 11  may include a NAD +  precursor molecule. For example, R 11  may include nicotinic acid, nicotinamide, or nicotinamide riboside. 
     R 11  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     The compound that shifts cellular metabolism from fatty acid oxidation may be represented by formula (VI): 
     
       
         
         
             
             
         
       
     
     in which: 
     at least one of positions A, B, C, D, E, and F is substituted with —(CH 2 CH 2 O) n H and n=1-15. 
     The compound may have a substitution at position F. For example, the compound may be represented by formula (IX), as shown above. 
     The compound that shifts cellular metabolism from fatty acid oxidation may be represented by formula (VII): 
     
       
         
           
             
               
                 
                   
                     A 
                     - 
                     C 
                   
                   , 
                 
               
               
                 
                   ( 
                   VIII 
                   ) 
                 
               
             
           
         
       
     
     in which A is a molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation, and C is a NAD +  precursor molecule. A and C may be covalently linked. 
     The molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be trimetazidine, etomoxir, perhexiline, a PPAR agonist, a malonyl CoA decarboxylase inhibitor, or dichloroacetate. 
     The molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be PEGylated with an ethylene glycol moiety. The molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may have multiple ethylene glycol moieties, such as one, two three, four, five, or more ethylene glycol moieties. The ethylene glycol moiety may be represented by (CH 2 CH 2 O) x , in which x=1-15. The ethylene glycol moiety may form a covalent linkage between the molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and the NAD +  precursor molecule. The ethylene glycol moiety may be separate from a covalent linkage between the molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and the NAD +  precursor molecule. The molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be a PEGylated form of trimetazidine. 
     The NAD +  precursor molecule may be nicotinic acid, nicotinamide, or nicotinamide riboside. 
     The compound of formula (VII) may include nicotinic acid that is covalently linked to a PEGylated form of trimetazidine. The nicotinic acid may be covalently linked via the PEGylated moiety, i.e., via an ethylene glycol linkage. The nicotinic acid may be covalently linked via the trimetazidine moiety. 
     The compound of formula (VII) may have a structure represented by formula (X), as shown above. 
     The compound that shifts cellular metabolism from fatty acid oxidation may be represented by formula (VIII): 
       A-L-C  (VIII),
 
     in which A is a molecule that molecule that shifts metabolism from fatty acid oxidation to glucose oxidation, L is a linker, and C is a NAD +  precursor molecule. A may be covalently linked to L, and L may be covalently linked to C. 
     The molecule that molecule that shifts metabolism from fatty acid oxidation to glucose oxidation, the linker, and the NAD +  precursor molecule may be as described above in relation to compounds of other formulas. 
     The compound of formula (VIII) may have a structure represented by formula (X), as shown above. 
     The compound that shifts cellular metabolism from fatty acid oxidation may be represented by formula (I): 
       A-L-B  (I),
 
     in which A is a molecule that molecule that shifts metabolism from fatty acid oxidation to glucose oxidation, L is a linker, and B is a compound that promotes mitochondrial respiration. 
     The molecule that shifts metabolism from fatty acid oxidation to glucose oxidation may be trimetazidine, etomoxir, perhexiline, a PPAR agonist, a malonyl CoA decarboxylase inhibitor, or dichloroacetate. 
     The compound that promotes mitochondrial respiration may be an intermediate of the citric acid cycle or a molecule that can be metabolized to enter the citric acid cycle. For example, the compound may be succinate, fumarate, malate, oxaloacetate, citrate, isocitrate, α-ketoglutarate, pyruvate, acetone, acetoacetic acid, β-hydroxybutpic acid, β-ketopentanoate, or β-hydroxypentanoate. 
     The linker may be any suitable linker that can be cleaved in vivo. The linker may be an alkoxy group. The linker may be polyethylene glycol of any length. Preferably, the linker is represented by (CH 2 CH 2 O) x , in which x=1-15. 
     The compound may include a NAD +  precursor molecule covalently linked to another component of the compound. The NAD +  precursor molecule may be nicotinic acid, nicotinamide, or nicotinamide riboside. The NAD +  precursor molecule may be attached to the molecule that molecule that shifts metabolism, the compound that promotes mitochondrial respiration, or the linker. The NAD +  precursor molecule may be attached to another component via an additional linker. Preferably, the NAD +  precursor molecule is attached to the compound that promotes mitochondrial respiration via a 1,3-propanediol linkage. 
     The compound of formula (I) may be represented by formula (II): 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     The compound of formula (I) may be represented by formula (III): 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     Any of the compounds described above may include one or more atoms that are enriched for an isotope. For example, the compounds may have one or more hydrogen atoms replaced with deuterium or tritium. The isotopically enriched atom or atoms may be located at any position within the compound. 
     The PDK inhibitor may be (R)-3.3.3-trifluoro-2-hydroxy-2-methyl propionamide, 2-chloroproprionate, 4,5-diarylisoxazole, anilide tertiary carbinol, aromatic DCA derivative, betulinic acid, CPI-613, dichloroacetate (DCA), a DCA-loaded tertiary amine, a DCA-oxaliplatin derivative, a dihydrolipoamide mimetic, a furan carboxylic acid, a hemoglobin-DCA conjugate (e.g., fusion molecule of 1 Hgb:12 DCAs), honokiol DCA, an inositol ester (e.g., inositol hexa(N-methylnicotinate-dichloroacetate), an inositol ionic complex (e.g., tetra(dichloroacetyl) gluconate), M77976, mitaplatin, mito-DCA (e.g., fusion molecule of 1 triphenylphosphonium cation: 3 DCA), N-(2-aminoethyl)-2(3-chloro-4-((4-isopropylbenzyl)oxy)phenyl)acetamide, phenylbutyrate, pyruvate, a pyruvate analog containing a phosphinate or phosphonate group, radicicol, R-lipoic acid, a tetrahydroisoquinoline, a thiophene carboxylic acid, or VER-246608. 
     In another aspect, the invention provides methods of treating a condition in a subject by providing a compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and a pyruvate dehydrogenase kinase (PDK) inhibitor. 
     The condition may be aneurysm, angina, atherosclerosis, brain ischemia, cancer, cardiac failure, cardiomyopathy, cardiovascular disease, cataracts, cerebral apoplexy, cerebral ischemia, cerebral vascular disease, congenital heart disease, coronary artery disease, coronary heart disease, diabetes, diabetic cardiomyopathy, diabetic complications, dyslipidemia, heart attack, heart failure, high blood pressure (hypertension), hyperglycemia, hyperlactacidemia, insulin resistance syndrome, ischemic heart disease, metabolic syndrome, mitochondrial disease, mitochondrial encephalomyopathy, myocardial ischemia, nephropathy, neuropathy, obesity, pericardial disease, peripheral arterial disease, pulmonary hypertension, retinopathy, rheumatic heart disease, stroke, transient ischemic attacks, valvular heart disease, or ventricular hypertrophy. 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and the PDK inhibitor may be provided in any suitable manner. They may be provided in a single composition. Alternatively, they may be provided in separate compositions. The agents may be provided simultaneously or sequentially. The agents may be provided at different intervals, with different frequency, or in different quantities. 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may include any of the elements and have any of the structural features described above in relation to combination therapies of the invention. 
     The PDK inhibitor may include any of the elements described above in relation to combination therapies of the invention. 
     In another aspect, the invention provides pharmaceutical compositions that include compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and a pyruvate dehydrogenase kinase (PDK) inhibitor. 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may include any of the elements and have any of the structural features described above in relation to combination therapies of the invention. 
     The PDK inhibitor may include any of the elements described above in relation to combination therapies of the invention. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a table summarizing the effects of various compounds on mitochondrial function. 
         FIG. 2  is a table summarizing the effects of nicotinamide on various mitochondrial functional parameters. 
         FIG. 3  is a series of graphs showing the effects of nicotinamide on oxygen consumption rate and reserve capacity. 
         FIG. 4  is a series of graphs showing the effects of nicotinamide on extracellular acidification rate. 
         FIG. 5  is a table summarizing the effects of a combination of trimetazidine and nicotinamide on various mitochondrial functional parameters. 
         FIG. 6  is a series of graphs showing the effects of a combination of trimetazidine and nicotinamide on oxygen consumption rate and reserve capacity. 
         FIG. 7  is a series of graphs showing the effects of a combination of trimetazidine and nicotinamide on extracellular acidification rate. 
         FIG. 8  is a table summarizing the effects of succinate on various mitochondrial functional parameters. 
         FIG. 9  is a series of graphs showing the effects of succinate on oxygen consumption rate and reserve capacity. 
         FIG. 10  is a series of graphs showing the effects of succinate on extracellular acidification rate. 
         FIG. 11  is a table summarizing the effects of compound CV-8816 on various mitochondrial functional parameters. 
         FIG. 12  is a series of graphs showing the effects of compound CV-8816 on oxygen consumption rate and reserve capacity. 
         FIG. 13  is a series of graphs showing the effects of compound CV-8816 on extracellular acidification rate. 
         FIG. 14  is a table summarizing the effects of compound CV-8814 on various mitochondrial functional parameters. 
         FIG. 15  is a series of graphs showing the effects of compound CV-8814 on oxygen consumption rate and reserve capacity. 
         FIG. 16  is a series of graphs showing the effects of compound CV-8814 on extracellular acidification rate. 
         FIG. 17  is a table summarizing the effects of trimetazidine on various mitochondrial functional parameters. 
         FIG. 18  is a series of graphs showing the effects of trimetazidine on oxygen consumption rate and reserve capacity. 
         FIG. 19  is a series of graphs showing the effects of trimetazidine on extracellular acidification rate. 
         FIG. 20  is a table summarizing the effects of compound CV-8815 on various mitochondrial functional parameters. 
         FIG. 21  is a series of graphs showing the effects of compound CV-8815 on oxygen consumption rate and reserve capacity. 
         FIG. 22  is a series of graphs showing the effects of compound CV-8815 on extracellular acidification rate. 
         FIG. 23  is a table summarizing the effects of a combination of succinate, nicotinamide, and trimetazidine on various mitochondrial functional parameters. 
         FIG. 24  is a series of graphs showing the effects of a combination of succinate, nicotinamide, and trimetazidine on oxygen consumption rate and reserve capacity. 
         FIG. 25  is a series of graphs showing the effects of a combination of succinate, nicotinamide, and trimetazidine on extracellular acidification rate. 
         FIG. 26  is a table summarizing the effects of a combination of trimetazidine analog 2 and nicotinamide on various mitochondrial functional parameters. 
         FIG. 27  is a series of graphs showing the effects of a combination of trimetazidine analog 2 and nicotinamide on oxygen consumption rate and reserve capacity. 
         FIG. 28  is a series of graphs showing the effects a combination of trimetazidine analog 2 and nicotinamide on extracellular acidification rate. 
         FIG. 29  is a table summarizing the effects of a combination of trimetazidine analog 1 and nicotinamide on various mitochondrial functional parameters. 
         FIG. 30  is a series of graphs showing the effects of a combination of trimetazidine analog 1 and nicotinamide on oxygen consumption rate and reserve capacity. 
         FIG. 31  is a series of graphs showing the effects of a combination of trimetazidine analog 1 and nicotinamide on extracellular acidification rate. 
         FIG. 32  is a table summarizing the effects of a combination of trimetazidine analog 3 and nicotinamide on various mitochondrial functional parameters. 
         FIG. 33  is a series of graphs showing the effects of a combination of trimetazidine analog 3 and nicotinamide on oxygen consumption rate and reserve capacity. 
         FIG. 34  is a series of graphs showing the effects of a combination of trimetazidine analog 3 and nicotinamide on extracellular acidification rate. 
         FIG. 35  is a table summarizing the effects of a combination of succinate and nicotinamide on various mitochondrial functional parameters. 
         FIG. 36  is a series of graphs showing the effects of a combination of succinate and nicotinamide on oxygen consumption rate and reserve capacity. 
         FIG. 37  is a series of graphs showing the effects of a combination of succinate and nicotinamide on extracellular acidification rate. 
         FIG. 38  is a schematic of the ischemia-reperfusion (IR) method used to analyze the effects of selected compositions on coronary flow. 
         FIG. 39  is a graph of coronary flow of after IR. 
         FIG. 40  is graph of left ventricular developed pressure (LVDP) after IR. 
         FIG. 41  shows images of TTC-stained heart slices after IR. 
         FIG. 42  is graph of infarct size after IR. 
         FIG. 43  is a schematic of the method used to analyze the effects of selected compositions on cardiac function. 
         FIG. 44  shows hearts from mice six weeks after transverse aortic constriction. 
         FIG. 45  is of graph of heart weight relative to body weight six weeks after transverse aortic constriction. 
         FIG. 46  is graph of heart weight six weeks after transverse aortic constriction. 
         FIG. 47  shows graphs of fractional shortening (FS) and ejection fraction (EF) at indicated time points after transverse aortic constriction. 
         FIG. 48  is a graph of left ventricular end-systolic diameter at indicated time points after transverse aortic constriction. 
         FIG. 49  is a graph of intraventricular septal dimension at indicated time points after transverse aortic constriction. 
         FIG. 50  is a graph of left ventricular mass at indicated time points after transverse aortic constriction. 
         FIG. 51  is a graph of isovolumic relaxation time at indicated time points after transverse aortic constriction. 
         FIG. 52  is a graph of the ratio peak velocity flow in early diastole vs. late diastole at indicated time points after transverse aortic constriction. 
         FIG. 53  is a graph of left ventricular developed pressure at six weeks after transverse aortic constriction. 
         FIG. 54  is a graph of the rate of left ventricle pressure rise at six weeks after transverse aortic constriction. 
         FIG. 55  is a graph showing levels of CV-8814 and trimetazidine after intravenous administration of CV-8834. 
         FIG. 56  is a graph showing levels of CV-8814 and trimetazidine after oral administration of CV-8834. 
         FIG. 57  is a graph showing levels of CV-8814 and trimetazidine after oral administration of CV-8834. 
         FIG. 58  is a graph showing levels of CV-8814 and trimetazidine after oral administration of CV-8834. 
         FIG. 59  is a graph showing levels of CV-8814 and trimetazidine after oral administration of CV-8834. 
         FIG. 60  is a graph showing levels of trimetazidine after oral administration of CV-8972 or intravenous administration of trimetazidine. 
         FIG. 61  is a graph showing levels of CV-8814 after oral administration of CV-8972 or intravenous administration of CV-8814. 
         FIG. 62  is a graph showing levels of CV-8814 after intravenous administration of CV-8834 or oral administration of CV-8834. 
         FIG. 63  is a graph showing levels of CV-8814 after intravenous administration of CV-8814 or oral administration of CV-8814. 
         FIG. 64  is a graph showing the HPLC elution profile of a batch of CV-8972. 
         FIG. 65  is a graph showing analysis of molecular species present in a batch of CV-8972. 
         FIG. 66  is a pair of graphs showing HPLC elution profiles of molecular species present in a batch of CV-8972. 
         FIG. 67  is a pair of graphs showing HPLC elution profiles of molecular species present in a batch of CV-8972. 
         FIG. 68  is a graph showing X-ray powder diffraction analysis of a batch of CV-8972. 
         FIG. 69  is a graph showing X-ray powder diffraction analysis of batches of CV-8972. 
         FIG. 70  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of a batch of CV-8972. 
         FIG. 71  is a graph showing dynamic vapor sorption (DVS) of a batch of CV-8972. 
         FIG. 72  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of a batch of CV-8972. 
         FIG. 73  is a graph showing dynamic vapor sorption (DVS) of a batch of CV-8972. 
         FIG. 74  is a graph showing X-ray powder diffraction analysis of samples of CV-8972. 
         FIG. 75  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of a batch of CV-8972. 
         FIG. 76  is a graph showing X-ray powder diffraction analysis of samples of CV-8972. 
         FIG. 77  is a graph showing X-ray powder diffraction analysis of samples of CV-8972. 
         FIG. 78  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of samples containing form A of CV-8972. 
         FIG. 79  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of a sample containing form A of CV-8972. 
     
    
    
     DETAILED DESCRIPTION 
     Regulation of cellular metabolism for the production of energy is critical to the progression and management of a variety of diseases, disorders, and conditions. Glucose oxidation and fatty acid oxidation are energy-producing metabolic pathways that compete with each other for substrates. In glucose oxidation, glucose is broken down to pyruvate via glycolysis in the cytosol of the cell. Pyruvate then enters the mitochondria, where it is converted to acetyl coenzyme A (acetyl-CoA) by the pyruvate dehydrogenase complex (PDC). In beta-oxidation of fatty acids, which occurs in the mitochondria, two-carbon units from long-chain fatty acids are sequentially converted to acetyl-CoA. 
     The remaining steps in energy production from oxidation of glucose or fatty acids are common to the two pathways. Acetyl-CoA is oxidized to carbon dioxide (CO 2 ) via the citric acid cycle (also called the tricarboxylic cycle, TCA cycle, and Krebs cycle), which results in the conversion of nicotinamide adenine dinucleotide (NAD + ) to its reduced form, NADH. NADH, in turn, drives the mitochondrial electron transport chain. The electron transport chain comprises a series of four mitochondrial membrane-bound complexes that transfer electrons via redox reactions and pump protons across the membrane to create a proton gradient. The redox reactions of the electron transport chain require molecular oxygen (O 2 ). Finally, the proton gradient enables another membrane-bound enzymatic complex to form high-energy ATP molecules, the primary intracellular molecule that drives energy-requiring reactions. 
     Because the PDC links glycolysis to the citric acid cycle, regulation of its activity plays a key gate-keeping function. When PDC activity is low, conversion of pyruvate to acetyl CoA is blocked, and pyruvate is instead converted to lactate in a reaction that also regenerates (NAD + ) from NADH. Thus, down-regulation of PDC activity uncouples glycolysis from the citric acid cycle and allows energy to be derived from glucose in oxygen-independent manner. Phosphorylation of the E1 pyruvate dehydrogenase subunit of PDC by pyruvate dehydrogenase kinase (PDK, also called PDHK and PDC kinase) deactivates the complex, and dephosphorylation of the same subunit by pyruvate dehydrogenase phosphatase (PDP) activates the complex. 
     Catabolism of fatty acids and glucose differ in ways that make each energy source advantageous under certain physiological conditions. Compared to glucose, fatty acids provide twice as much ATP per mass unit and thus are more efficient than glucose as a source of stored energy. However, ATP can be produced faster from glucose than from fatty acids. In addition, fatty acid oxidation requires more oxygen than does glucose oxidation. Glucose oxidation includes both glycolysis, which yields ATP in a series of oxygen-independent reactions, and post-glycolytic reactions, i.e., pyruvate decarboxylation and the citric acid cycle, which require oxygen and generate the majority of ATP produced by glucose oxidation. Consequently, in physiological conditions in which energy is needed rapidly and/or oxygen is scarce, such as in muscles during intensive exercise or in ischemic tissue, catabolism of glucose is preferred. 
     Cellular regulation of metabolic pathways for energy production is critical in a variety of diseases and conditions. For example, in cardiovascular conditions that lead to reduced blood flow to tissues, oxygen levels are insufficient to support fatty acid oxidation. Therefore, providing agents that promote glucose oxidation can provide therapeutic benefits. For example, in cases of angina, restoration of energy production in cardiac tissue can reduce the risk of myocardial infarction. In some cases of diabetes and obesity, overexpression of PDK4, which encodes an isoform of PDK, inhibits activity of the PDC and impairs glucose oxidation. Genes encoding other PDK isoforms, such as PDK1 and PDK3, are overexpressed in certain cancers. Without wishing to be bound by any particular theory, is thought that inhibition of PDC allows tumor cells to rely exclusively on glycolysis for energy production and avoid apoptotic signals that would otherwise be generated by cells in hypoxic conditions. Survival under hypoxic conditions is a key adaptation that permits metastatic invasion of tumor cells into other tissues. 
     The invention provides combination therapies that correct metabolic defects, such as those described above, by promoting glucose oxidation. The combination therapies include an agent that shifts cellular metabolism from fatty acid oxidation to glucose oxidation, such as trimetazidine or an analog, derivative, or prodrug thereof, and an inhibitor of pyruvate dehydrogenase kinase. The first agent promotes the use of glucose as an energy source, and the second agent shunts the pyruvate produced from glycolysis into the citric acid cycle. By driving complete oxidation of glucose, the therapies ensure that the energy yield from glucose catabolism is maximized and that mitochondria produce apoptotic signals under appropriate conditions. The combination therapies may also include a NAD +  precursor molecule. The invention includes compositions containing the therapeutic combinations of the invention and methods of treating conditions using such combinations. 
     Compositions 
     The invention includes combination therapies that include a compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and an inhibitor of pyruvate dehydrogenase kinase. Compounds that shift cellular metabolism from fatty acid oxidation to glucose oxidation are described in, for example, International Patent Publication No. WO 2018/236745, the contents of which are incorporated herein by reference. 
     Compounds that Shift Cellular Metabolism from Fatty Acid Oxidation to Glucose Oxidation 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be represented by formula (I): 
     
       
         
           
             
               
                 
                   
                     A 
                     - 
                     L 
                     - 
                     B 
                   
                   , 
                 
               
               
                 
                   ( 
                   I 
                   ) 
                 
               
             
           
         
       
     
     in which A is a molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation, L is a linker, and B is a compound that promotes mitochondrial respiration. 
     Component A may be any suitable molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation. Such compounds can be classified based on their mechanism of action. See Fillmore, N., et al., Mitochondrial fatty acid oxidation alterations in heart failure, ischemic heart disease and diabetic cardiomyopathy, Brit. J. Pharmacol. 171:2080-2090 (2014), incorporated herein by reference. 
     One class of glucose-shifting compounds includes compounds that inhibit fatty acid oxidation directly. Compounds in this class include inhibitors of malonyl CoA decarboxylase (MCD), carnitine palmitoyl transferase 1 (CPT-1), or mitochondrial fatty acid oxidation. Mitochondrial fatty acid oxidation inhibitors include trimetazidine and other compounds described in International Patent Publication No. WO 2002/064,576, the contents of which is incorporated herein by reference. Trimetazidine binds to distinct sites on the inner and outer mitochondrial membranes and affects both ion permeability and metabolic function of mitochondria. Morin, D., et al., Evidence for the existence of [ 3 H]-trimetazidine binding sites involved in the regulation of the mitochondrial permeability transition pore, Brit. J. Pharmacol. 123:1385-1394 (1998), incorporated herein by reference. MCD inhibitors include CBM-301106, CBM-300864, CBM-301940, 5-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)-4,5-dihydroisoxazole-3-carboxamides, methyl 5-(N-(4-(1,1,1,3,3,3-hexafluoro-2-hydroxypropan-2-yl)phenyl)morpholine-4-carboxamido)pentanoate, and other compounds described in Chung, J. F., et al., Discovery of Potent and Orally Available Malonyl-CoA Decarboxylase Inhibitors as Cardioprotective Agents, J. Med. Chem. 49:4055-4058 (2006); Cheng J. F. et al., Synthesis and structure-activity relationship of small-molecule malonyl coenzyme A decarboxylase inhibitors, J. Med. Chem. 49:1517-1525 (2006); U.S. Patent Publication No. 2004/0082564; and International Patent Publication No. WO 2002/058,698, the contents of which are incorporated herein by reference. CPT-1 inhibitors include oxfenicine, perhexiline, etomoxir, and other compounds described in International Patent Publication Nos. WO 2015/018,660; WO 2008/109,991; WO 2009/015,485; and WO 2009/156479; and U.S. Patent Publication No. 2011/0212072, the contents of which are incorporated herein by reference. 
     Another class of glucose-shifting compounds includes compounds that stimulate glucose oxidation directly. Examples of such compounds are described in U.S. Patent Publication No. 2003/0191182; International Patent Publication No. WO 2006/117,686; U.S. Pat. No. 8,202,901, the content of each of which are incorporated herein by reference. 
     Another class of glucose-shifting compounds includes compounds that decrease the level of circulating fatty acids that supply the heart. Examples of such compounds include agonists of PPARα and PPARγ, including fibrate drugs, such as clofibrate, gemfibrozil, ciprofibrate, bezafibrate, and fenofibrate, and thiazolidinediones, GW-9662, and other compounds described in U.S. Pat. No. 9,096,538, which is incorporated herein by reference. 
     Component L may be any suitable linker. Preferably, the linker can be cleaved in vivo to release components A and B. The linker may be an alkoxy group. The linker may be polyethylene glycol of any length. The linker may be represented by (CH 2 CH 2 O) x , in which x=1-15 or (CH 2 CH 2 O) x , in which x=1-3. Other suitable linkers include 1,3-propanediol, diazo linkers, phosphoramidite linkers, disulfide linkers, cleavable peptides, iminodiacetic acid linkers, thioether linkers, and other linkers described in Leriche, G., et al., Cleavable linkers in chemical biology, Bioorg. Med. Chem. 20:571-582 (2012); International Patent Publication No. WO 1995/000,165; and U.S. Pat. No. 8,461,117, the contents of which are incorporated herein by reference. 
     Component B may be any compound that promotes mitochondrial respiration. For example, component B may be an intermediate of the citric acid cycle or a molecule that can be metabolized to enter the citric acid cycle, such as succinate, fumarate, malate, oxaloacetate, citrate, isocitrate, α-ketoglutarate, pyruvate, acetone, acetoacetic acid, β-hydroxybutpic acid, β-ketopentanoate, or β-hydroxypentanoate. Intermediates of the citric acid cycle may become depleted if these molecules are used for biosynthetic purposes, resulting in inefficient generation of ATP from the citric acid cycle. However, due to the anaplerotic effect, providing one intermediate of the citric acid cycle leads to restoration of all intermediates as the cycle turns. Thus, intermediates of the citric acid cycle can promote mitochondrial respiration. 
     The compound may include a NAD +  precursor molecule. NAD +  is an important oxidizing agent that acts as a coenzyme in multiple reactions of the citric acid cycle. In these reactions, NAD +  is reduced to NADH. Conversely, NADH is oxidized back to NAD +  when it donates electrons to mitochondrial electron transport chain. In humans, NAD +  can be synthesized de novo from tryptophan, but not in quantities sufficient to meet metabolic demands. Consequently, NAD +  is also synthesized via a salvage pathway, which uses precursors that must be supplied from the diet. Among the precursors used by the salvage pathway for NAD +  synthesis are nicotinic acid, nicotinamide, and nicotinamide riboside. By providing a NAD +  precursor, such as nicotinic acid, nicotinamide, or nicotinamide riboside, the compound facilitates NAD +  synthesis. 
     The inclusion of a NAD +  precursor allows the compounds to stimulate energy production in cardiac mitochondria in multiple ways. First, component A shifts cellular metabolism from fatty acid oxidation to glucose oxidation, which is inherently more efficient. Next, component B ensures that the intermediates of the citric acid cycle are present at adequate levels and do not become depleted or limiting. As a result, glucose-derived acetyl CoA is efficiently oxidized. Finally, the NAD +  precursor provides an essential coenzyme that cycles between oxidized and reduced forms to promote respiration. In the oxidized form, NAD +  drives reactions of the citric acid cycle. In the reduced form, NADH promotes electron transport to create a proton gradient that enables ATP synthesis. Consequently, the chemical potential resulting from oxidation of acetyl CoA is efficiently converted to ATP that can be used for various cellular functions. 
     The NAD +  precursor molecule may be covalently attached to the compound in any suitable manner. For example, it may be linked to A, L, or B, and it may be attached directly or via another linker. Preferably, it is attached via a linker that can be cleaved in vivo. The NAD +  precursor molecule may be attached via a 1,3-propanediol linkage. 
     The compound may be covalently attached to one or more molecules of polyethylene glycol (PEG), i.e., the compound may be PEGylated. In many instances, PEGylation of molecules reduces their immunogenicity, which prevents the molecules from being cleared from the body and allows them to remain in circulation longer. The compound may contain a PEG polymer of any size. For example, the PEG polymer may have from 1-500 (CH 2 CH 2 O) units. The PEG polymer may have any suitable geometry, such as a straight chain, branched chain, star configuration, or comb configuration. The compound may be PEGylated at any site. For example, the compound may be PEGylated on component A, component B, component L, or, if present, the NAD +  precursor. The compound may be PEGylated at multiple sites. For a compound PEGylated at multiple sites, the various PEG polymers may be of the same or different size and of the same or different configuration. 
     The compound may be a PEGylated form of trimetazidine. For example, the compound may be represented by formula (VI): 
     
       
         
         
             
             
         
       
     
     in which one or more of the carbon atoms at positions A, B, C, D, and E and/or the nitrogen atom at position F are substituted with —(CH 2 CH 2 O) n H and n=1-15. The carbon atoms at positions A, B, C, D, and E may have two PEG substituents. In molecules that have multiple PEG chains, the different PEG chains may have the same or different length. 
     The compounds of formula (I) may be represented by formula (II): 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     The compounds of formula (I) may be represented by formula (III): 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be represented by formula (IV): 
     
       
         
         
             
             
         
       
     
     in which R 1 , R 2 , and R 3  are independently H or a (C 1 -C 4 )alkyl group; R 4  and R 5  together are ═O, —O(CH 2 ) m O—, or —(CH 2 ) m —, in which m=2-4, or R 4  is H and R 5  is OR 14 , SR 14 , or (CH 2 CH 2 O) n H, in which R 14  is H or a (C 1 -C 4 )alkyl group and n=1-15; and R 6  is a single or multi-ring structure optionally substituted at one or more ring positions by a heteroatom, in which each ring position optionally comprises one or more substituents. 
     R 6  may be a single or multi-ring structure of any size. For example, the structure may contain 3-22 atoms, not including hydrogen atoms bonded to atoms in ring positions. The structure may include one or more alkyl, alkenyl, or aromatic rings. The structure may include one or more heteroatoms, i.e., atoms other than carbon. For example, the heteroatom may be oxygen, nitrogen, or sulfur, or phosphorus. 
     One or more ring position of R 6  may include a substituent that includes a compound that promotes mitochondrial respiration, as described above in relation to component B of formula (I). The substituent may include a linker, as described above in relation to component L of formula (I). The substituent may include a NAD +  precursor molecule, as described above in relation to compounds of formula (I). 
     The substituent on a ring position of R 6  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     The substituent on a ring position of R 6  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     R 6  may be 
     
       
         
         
             
             
         
       
     
     For some compounds that include trimetazidine prodrugs, analogs, derivatives, it is advantageous to have the trimetazidine moiety substituted with a single ethylene glycol moiety. Thus, compositions of the invention may include compounds of formulas (I) and (VIII) that contain linkers in which x=1, compounds of formulas (II) and (III) in which y=1, compounds of formula (V) in which z=1, compounds of formula (VI) in which n=1, and compounds of formula (VII) in which A is linked to C via a single ethylene glycol moiety. Without wishing to be bound by theory, the attachment of a single ethylene glycol moiety to the trimetazidine moiety may improve the bioavailability of trimetazidine. 
     The compound of formula (IV) may have a structure represented by formula (IX) or formula (X): 
     
       
         
         
             
             
         
       
     
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be represented by formula (V): 
     
       
         
         
             
             
         
       
     
     in which R 1 , R 2 , and R 3  are independently H or a (C 1 -C 4 )alkyl group; R 4  and R 8  together are ═O, —O(CH 2 ) m O—, or —(CH 2 ) m —, in which m=2-4, or R 4  is H and R 8  is H, OR 14 , SR 14 , or (CH 2 CH 2 O) n H, in which R 14  is H or a (C 1 -C 4 )alkyl group and n=1-15; R 9 , R 10 , R 12 , and R 13  are independently H or (CH 2 CH 2 O) z H, in which z=1-15; and R 11  comprises a compound that promotes mitochondrial respiration, as described above in relation to component B of formula (I). R 11  may include a linker, as described above in relation to component L of formula (I). R 11  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3.
 
R 11  may include a NAD +  precursor molecule, as described above in relation to compounds of formula (I).
 
     R 11  may be 
     
       
         
         
             
             
         
       
     
     in which y=1-3. 
     In some embodiments described above, the compound includes multiple active agents joined by linkers in a single molecule. It may be advantageous to deliver multiple active agents as components of a single molecule. Without wishing to be bound by a particular theory, there are several reasons why co-delivery of active agents in a single molecule may be advantageous. One possibility is that a single large molecule may have reduced side effects compared to the component agents. Free trimetazidine causes symptoms similar to those in Parkinson&#39;s disease in a fraction of patients. However, when trimetazidine is derivatized to include other components, such as succinate, the molecule is bulkier and may not be able to access sites where free trimetazidine can causes unintended effects. Trimetazidine derivatized as described above is also more hydrophilic and thus may be less likely to cross the blood-brain barrier to cause neurological effects. Another possibility is that modification of trimetazidine may alter its pharmacokinetic properties. Because the derivatized molecule is metabolized to produce the active agent, the active agent is released gradually. Consequently, levels of the active agent in the body may not reach peaks as high as when a comparable amount is administered in a single bolus. Another possibility is that less of each active agent, such as trimetazidine, is required because the compositions of the invention may include compounds that have multiple active agents. For example, trimetazidine shifts metabolism from fatty acid oxidation to glucose oxidation, and succinate improves mitochondrial respiration generally. Thus, a compound that provides both agents stimulates a larger increase in glucose-driven ATP production for a given amount of trimetazidine than does a compound that delivers trimetazidine alone. 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be represented by formula (VII): 
       A-C  (VII),
 
     in which A is a molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation, and C is a NAD +  precursor molecule. A and C may be covalently linked. 
     The molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be PEGylated with an ethylene glycol moiety. The molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may have multiple ethylene glycol moieties, such as one, two three, four, five, or more ethylene glycol moieties. The ethylene glycol moiety may be represented by (CH 2 CH 2 O) x , in which x=1-15. The ethylene glycol moiety may form a covalent linkage between the molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and the NAD +  precursor molecule. The ethylene glycol moiety may be separate from a covalent linkage between the molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and the NAD +  precursor molecule. 
     The compound of formula (VII) may include nicotinic acid that is covalently linked to a PEGylated form of trimetazidine. The nicotinic acid may be covalently linked via a PEGylated moiety, i.e., via an ethylene glycol linkage. The nicotinic acid may be covalently linked via the trimetazidine moiety. 
     The compound that shifts cellular metabolism from fatty acid oxidation to glucose oxidation may be represented by formula (VIII): 
       A-L-C  (VIII),
 
     in which A is a molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation, L is a linker, and C is a NAD +  precursor molecule. A may be covalently linked to L, and L may be covalently linked to C. 
     The molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation, the linker, and the NAD +  precursor molecule may be as described above in relation to compounds of other formulas. 
     The compounds may be provided as co-crystals with other compounds. Co-crystals are crystalline materials composed of two or more different molecules in the same crystal lattice. The different molecules may be neutral and interact non-ionically within the lattice. Co-crystals may include one or more of the compounds described above and one or more other molecules that stimulate mitochondrial respiration or serve as NAD  + precursors. For example, a co-crystal may include any of the following combinations: (1) a molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and (2) a NAD +  precursor molecule; (1) a compound that promotes mitochondrial respiration and (2) a NAD +  precursor molecule; (1) a molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and (2) a compound that promotes mitochondrial respiration; (1) a molecule comprising a molecule that shifts cellular metabolism from fatty acid oxidation to glucose oxidation covalently linked to a compound that promotes mitochondrial respiration and (2) a NAD +  precursor molecule. In specific embodiments, a co-crystal may include (1) a compound of formula (I), (III), (IV), or (V) and (2) nicotinic acid, nicotinamide, or nicotinamide riboside. 
     The compounds may include one or more atoms that are enriched for an isotope. For example, the compounds may have one or more hydrogen atoms replaced with deuterium or tritium. Isotopic substitution or enrichment may occur at carbon, sulfur, or phosphorus, or other atoms. The compounds may be isotopically substituted or enriched for a given atom at one or more positions within the compound, or the compounds may be isotopically substituted or enriched at all instances of a given atom within the compound. 
     Inhibitors of PDK 
     The PDK inhibitor may be any agent that inhibits one or more isoforms of pyruvate dehydrogenase kinase. The PDK inhibitor may be any suitable class of molecule. For example and without limitation, the PDK inhibitor may be a small molecule, protein, peptide, polypeptide, nucleic acid (e.g., RNA, siRNA, shRNA, miRNA, mRNA, DNA, nucleic acid with one or more modified nucleotides, etc.), or combination thereof. For example and without limitation, the PDK inhibitor may be (R)-3.3.3-trifluoro-2-hydroxy-2-methyl propionamide, 2-chloroproprionate, 4,5-diarylisoxazole, anilide tertiary carbinol, aromatic DCA derivative, betulinic acid, CPI-613, dichloroacetate (DCA), a DCA-loaded tertiary amine, a DCA-oxaliplatin derivative, a dihydrolipoamide mimetic, a furan carboxylic acid, a hemoglobin-DCA conjugate (e.g., fusion molecule of 1 Hgb:12 DCAs), honokiol DCA, an inositol ester (e.g., inositol hexa(N-methylnicotinate-dichloroacetate), an inositol ionic complex (e.g., tetra(dichloroacetyl) gluconate), M77976, mitaplatin, mito-DCA (e.g., fusion molecule of 1 triphenylphosphonium cation: 3 DCA), N-(2-aminoethyl)-2(3-chloro-4-(4-isopropylbenzyl)oxy)phenyl)acetamide, phenylbutyrate, pyruvate, a pyruvate analog containing a phosphinate or phosphonate group, radicicol, R-lipoic acid, a tetrahydroisoquinoline, a thiophene carboxylic acid, or VER-246608. PDK inhibitors are known in the art and described in, for example, U.S. Patent Publication No. 2017/0001958; U.S. Pat. No. 8,871,934; International Patent Publication Nos. WO 2015/040,424 and WO 2017/167,676; and Peter W. Stacpoole, Therapeutic Targeting of the Pyruvate Dehydrogenase Complex/Pyruvate Dehydrogenase Kinase (PDC/PDK) Axis in Cancer, JNCI: Journal of the National Cancer Institute, Volume 109, Issue 11, 1 Nov. 2017, djx071, doi: 10.1093/jnci/djx071, the contents of each of which are incorporated herein by reference. 
     Formulations 
     The invention provides pharmaceutical compositions containing one or more of the compounds described above. A pharmaceutical composition containing the compounds may be in a form suitable for oral use, for example, as tablets, troches, lozenges, fast-melts, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs. Compositions intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the compounds in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration in the stomach and absorption lower down in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. They may also be coated to form osmotic therapeutic tablets for control release by the techniques described in U.S. Pat. Nos. 4,256,108, 4,166,452 and 4,265,874, the contents of which are incorporated herein by reference. Preparation and administration of compounds is discussed in U.S. Pat. No. 6,214,841 and U.S. Pub. 2003/0232877, the contents of which are incorporated by reference herein in their entirety. 
     Formulations for oral use may also be presented as hard gelatin capsules in which the compounds are mixed with an inert solid diluent, for example calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the compounds are mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil. 
     An alternative oral formulation, where control of gastrointestinal tract hydrolysis of the compound is sought, can be achieved using a controlled-release formulation, where a compound is encapsulated in an enteric coating. 
     Aqueous suspensions may contain the compounds in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents such as a naturally occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such a polyoxyethylene with partial esters derived from fatty acids and hexitol anhydrides, for example polyoxyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin. 
     Oily suspensions may be formulated by suspending the compounds in a vegetable oil, for example, arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid. 
     Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the compounds in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified, for example sweetening, flavoring and coloring agents, may also be present. 
     The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally occurring phosphatides, for example soya bean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents. 
     Syrups and elixirs may be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, and agents for flavoring and/or coloring. The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be in a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer&#39;s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. 
     The compositions of the invention are useful for improving cardiac efficiency. A variety of definitions of cardiac efficiency exist in the medical literature. See, e.g.. Schipke, J. D. Cardiac efficiency, Basic Res. Cardiol. 89:207-40 (1994); and Gibbs, C. L. and Barclay, C. J. Cardiac efficiency, Cardiovasc. Res. 30:627-634 (1995), incorporated herein by reference. One definition of cardiac mechanical efficiency is the ratio of external cardiac power to cardiac energy expenditure by the left ventricle. See Lopaschuk G. D., et al., Myocardial Fatty Acid Metabolism in Health and Disease, Phys. Rev. 90:207-258 (2010), incorporated herein by reference. Another definition is the ratio between stroke work and oxygen consumption, which ranges from 20-25% in the normal human heart. Visser, F., Measuring cardiac efficiency: is it useful? Hear Metab. 39:3-4 (2008), incorporated herein by reference. Another definition is the ratio of the stroke volume to mean arterial blood pressure. Any suitable definition of cardiac efficiency may be used to measure the effects of compositions of the invention 
     The compositions of the invention may contain an agent that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and an inhibitor of PDK in a single formulation. Alternatively, the compositions of the invention may contain an agent that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and an inhibitor of PDK in separate formulations. The compositions may contain a NAD +  precursor molecule in a formulation that contains an agent that shifts cellular metabolism from fatty acid oxidation to glucose oxidation and/or an inhibitor of PDK, or the NAD +  precursor molecule may be provided in a separate formulation. 
     Methods of Treating Diseases, Disorders, and Conditions 
     The invention also provides methods of treating diseases, disorders, and conditions using the combination therapies described herein. The combination therapies are useful for treating any disease, disorder, or condition for which a shift in cellular metabolism from fatty acid oxidation to glucose oxidation would be advantageous. 
     The combination therapies are useful for treating or preventing diseases relating to glucose utilization, such as diabetes (e.g., type 1 diabetes, type 2 diabetes), insulin resistance syndrome, metabolic syndrome, hyperglycemia, and hyperlactacidemia. The combination therapies may also be used to treat complications of the aforementioned disorders, such as neuropathy, retinopathy, nephropathy, and cataracts. 
     The combination therapies may be used to treat or prevent diseases, disorders, or conditions associated with a limited supply of energy substrates to tissues, such as cardiac failure, cardiomyopathy, myocardial ischemia, dyslipidemia, atherosclerosis, and cerebral ischemia. 
     The combination therapies may be used to treat or prevent diseases, disorders, or conditions associated with mitochondrial dysfunction, such as mitochondrial disease and mitochondrial encephalomyopathy. 
     Other categories of diseases that can be treated or prevented with combination therapies of the invention include cardiovascular disease, cancer, and pulmonary hypertension. 
     For example and without limitation, the combination therapies may be used to treat or prevent aneurysm, angina, atherosclerosis, brain ischemia, cancer, cardiac failure, cardiomyopathy, cardiovascular disease, cataracts, cerebral apoplexy, cerebral ischemia, cerebral vascular disease, congenital heart disease, coronary artery disease, coronary heart disease, diabetes, diabetic cardiomyopathy, diabetic complications, dyslipidemia, heart attack, heart failure, high blood pressure (hypertension), hyperglycemia, hyperlactacidemia, insulin resistance syndrome, ischemic heart disease, metabolic syndrome, mitochondrial disease, mitochondrial encephalomyopathy, myocardial ischemia, nephropathy, neuropathy, obesity, pericardial disease, peripheral arterial disease, pulmonary hypertension, retinopathy, rheumatic heart disease, stroke, transient ischemic attacks, valvular heart disease, or ventricular hypertrophy. 
     The compositions may be provided by any suitable route of administration. For example and without limitation, the compositions may be administered buccally, by injection, dermally, enterally, intraarterially, intravenously, nasally, orally, parenterally, pulmonarily, rectally, subcutaneously, topically, transdermally, or with or on an implantable medical device (e.g., stent or drug-eluting stent or balloon equivalents). 
     EXAMPLES 
     Protocol 
     The effects of selected compounds on mitochondrial function were analyzed. HepG2 cells were dosed with test compound and in real time the extracellular oxygen levels and pH were measured using the XFe96 flux analyzer (Seahorse Biosciences). XFe Technology uses solid-state sensors to simultaneously measure both oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) to determine effects on oxidative phosphorylation (OXPHOS) and glycolysis simultaneously. The cells were then subjected to sequential exposure to various inhibitors of mitochondrial function to assess cellular metabolism. 
     Data Interpretation 
     A compound was identified as positive mitochondrial-active compound when it caused a change in oxygen consumption rate (OCR) or extracellular acidification rate (ECAR) in the absence of cytotoxicity. Cytotoxicity was determined when both OXPHOS (OCR) and glycolysis (ECAR) were inhibited. 
     Definition of Mitochondrial Parameters 
     Oxygen consumption rate (OCR) is a measurement of oxygen content in extracellular media. Changes in OCR indicate effects on mitochondrial function and can be bi-directional. A decrease is due to an inhibition of mitochondrial respiration, while an increase may indicate an uncoupler, in which respiration is not linked to energy production. 
     
       
         
           
             OCR 
             = 
             
               
                 
                   compound 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
                 - 
                 
                   non 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   mitochondrial 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
               
               
                 
                   basal 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
                 - 
                 
                   non 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   mitochondrial 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
               
             
           
         
       
     
     Extracellular acidification rate (ECAR) is the measurement of extracellular proton concentration (pH). An increase in signal means an increase in rate in number of pH ions (thus decreasing pH value) and seen as an increase in glycolysis. ECAR is expressed as a fraction of basal control (rate prior to addition of compound). 
     
       
         
           
             
               reserve 
               ⁢ 
               
                   
               
               ⁢ 
               capacity 
             
             = 
             
               
                 
                   FCCP 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
                 - 
                 
                   non 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   mitochondrial 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
               
               
                 
                   basal 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
                 - 
                 
                   non 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   mitochondrial 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   OCR 
                 
               
             
           
         
       
     
     Reserve capacity is the measured ability of cells to respond to an increase in energy demand. A reduction indicates mitochondrial dysfunction. This measurement demonstrates how close to the bioenergetic limit the cell is. 
     
       
         
           
             ECAR 
             = 
             
               
                 compound 
                 ⁢ 
                 
                     
                 
                 ⁢ 
                 ECAR 
               
               
                 basal 
                 ⁢ 
                 
                     
                 
                 ⁢ 
                 ECAR 
               
             
           
         
       
     
     Mitochondrial Stress Test 
     A series of compounds were added sequentially to the cells to assess a bioenergetics profile, effects of test compounds on parameters such as proton leak, and reserve capacity. This can be used to assist in understanding potential mechanisms of mitochondrial toxicity. The following compounds were added in order: (1) oligomycin, (2) FCCP, and (3) rotenone and antimycin A. 
     Oligomycin is a known inhibitor of ATP synthase and prevents the formation of ATP. Oligomycin treatment provides a measurement of the amount of oxygen consumption related to ATP production and ATP turnover. The addition of oligomycin results in a decrease in OCR under normal conditions, and residual OCR is related to the natural proton leak. 
     FCCP is a protonophore and is a known uncoupler of oxygen consumption from ATP production. FCCP treatment allows the maximum achievable transfer of electrons and oxygen consumption rate and provides a measurement of reserve capacity. 
     Rotenone and antimycin A are known inhibitors of complex I and III of the electron transport chain, respectively. Treatment with these compounds inhibits electron transport completely, and any residual oxygen consumption is due to non-mitochondrial activity via oxygen requiring enzymes. 
     Definition of Mechanisms 
     An electron transport chain inhibitor is an inhibitor of mitochondrial respiration that causes an increase in glycolysis as an adaptive response (e.g. decrease OCR and increase in ECAR). 
     The inhibition of oxygen consumption may also be due to reduced substrate availability (e.g. glucose, fatty acids, glutamine, pyruvate), for example, via transporter inhibition. Compounds that reduce the availability of substrates are substrate inhibitors. A substrate inhibitor does not result in an increase in glycolysis (e.g. OCR decrease, no response in ECAR). 
     Compounds that inhibit the coupling of the oxidation process from ATP production are known as uncouplers. These result in an increase in mitochondrial respiration (OCR) but inhibition of ATP production. 
       FIG. 1  is a table summarizing the effects of various compounds on mitochondrial function. 
       FIG. 2  is a table summarizing the effects of nicotinamide on various mitochondrial functional parameters. 
       FIG. 3  is a series of graphs showing the effects of nicotinamide on oxygen consumption rate and reserve capacity. 
       FIG. 4  is a series of graphs showing the effects of nicotinamide on extracellular acidification rate. 
       FIG. 5  is a table summarizing the effects of a combination of trimetazidine and nicotinamide on various mitochondrial functional parameters. 
       FIG. 6  is a series of graphs showing the effects of a combination of trimetazidine and nicotinamide on oxygen consumption rate and reserve capacity. 
       FIG. 7  is a series of graphs showing the effects of a combination of trimetazidine and nicotinamide on extracellular acidification rate. 
       FIG. 8  is a table summarizing the effects of succinate on various mitochondrial functional parameters. 
       FIG. 9  is a series of graphs showing the effects of succinate on oxygen consumption rate and reserve capacity. 
       FIG. 10  is a series of graphs showing the effects of succinate on extracellular acidification rate. 
       FIG. 11  is a table summarizing the effects of compound CV-8816 on various mitochondrial functional parameters. 
       FIG. 12  is a series of graphs showing the effects of compound CV-8816 on oxygen consumption rate and reserve capacity. 
       FIG. 13  is a series of graphs showing the effects of compound CV-8816 on extracellular acidification rate. 
       FIG. 14  is a table summarizing the effects of compound CV-8814 on various mitochondrial functional parameters. 
       FIG. 15  is a series of graphs showing the effects of compound CV-8814 on oxygen consumption rate and reserve capacity. 
       FIG. 16  is a series of graphs showing the effects of compound CV-8814 on extracellular acidification rate. 
       FIG. 17  is a table summarizing the effects of trimetazidine on various mitochondrial functional parameters. 
       FIG. 18  is a series of graphs showing the effects of trimetazidine on oxygen consumption rate and reserve capacity. 
       FIG. 19  is a series of graphs showing the effects of trimetazidine on extracellular acidification rate. 
       FIG. 20  is a table summarizing the effects of compound CV-8815 on various mitochondrial functional parameters. 
       FIG. 21  is a series of graphs showing the effects of compound CV-8815 on oxygen consumption rate and reserve capacity. 
       FIG. 22  is a series of graphs showing the effects of compound CV-8815 on extracellular acidification rate. 
       FIG. 23  is a table summarizing the effects of a combination of succinate, nicotinamide, and trimetazidine on various mitochondrial functional parameters. 
       FIG. 24  is a series of graphs showing the effects of a combination of succinate, nicotinamide, and trimetazidine on oxygen consumption rate and reserve capacity. 
       FIG. 25  is a series of graphs showing the effects of a combination of succinate, nicotinamide, and trimetazidine on extracellular acidification rate. 
       FIG. 26  is a table summarizing the effects of a combination of trimetazidine analog 2 and nicotinamide on various mitochondrial functional parameters. 
       FIG. 27  is a series of graphs showing the effects of a combination of trimetazidine analog 2 and nicotinamide on oxygen consumption rate and reserve capacity. 
       FIG. 28  is a series of graphs showing the effects a combination of trimetazidine analog 2 and nicotinamide on extracellular acidification rate. 
       FIG. 29  is a table summarizing the effects of a combination of trimetazidine analog 1 and nicotinamide on various mitochondrial functional parameters. 
       FIG. 30  is a series of graphs showing the effects of a combination of trimetazidine analog 1 and nicotinamide on oxygen consumption rate and reserve capacity. 
       FIG. 31  is a series of graphs showing the effects of a combination of trimetazidine analog 1 and nicotinamide on extracellular acidification rate. 
       FIG. 32  is a table summarizing the effects of a combination of trimetazidine analog 3 and nicotinamide on various mitochondrial functional parameters. 
       FIG. 33  is a series of graphs showing the effects of a combination of trimetazidine analog 3 and nicotinamide on oxygen consumption rate and reserve capacity. 
       FIG. 34  is a series of graphs showing the effects of a combination of trimetazidine analog 3 and nicotinamide on extracellular acidification rate. 
       FIG. 35  is a table summarizing the effects of a combination of succinate and nicotinamide on various mitochondrial functional parameters. 
       FIG. 36  is a series of graphs showing the effects of a combination of succinate and nicotinamide on oxygen consumption rate and reserve capacity. 
       FIG. 37  is a series of graphs showing the effects of a combination of succinate and nicotinamide on extracellular acidification rate. 
     Effect of Compositions on Coronary Flow, Cardiac Function, and Infarct Size 
     The effect of compositions on the coronary flow, cardiac function, and infarct size was analyzed. 
       FIG. 38  is a schematic of the ischemia-reperfusion (IR) method used to analyze the effects of selected compositions on coronary flow, cardiac function, and infarct size. At time 0, mice were given (1) 20 μM trimetazidine (TMZ), (2) 2 μM each of trimetazidine, nicotinamide, and succinate (TNF), (3) 20 μM each of trimetazidine, nicotinamide, and succinate (TNS), or (4) the delivery vehicle (CON). At 20 minutes, ischemia was induced, and coronary flow was analyzed. At 50 minutes, reperfusion was initiated to restore blood flow. At 170 minutes, coronary flow and cardiac function was analyzed, and then the hearts were preserved, sectioned, and infarct size was measured by triphenyltetrazolium chloride (TTC) staining. 
       FIG. 39  is a graph of coronary flow of after IR. Data is expressed as ratio cardiac flow at 170 minutes to cardiac flow at 20 minutes. TNS treatment preserved coronary flow after IR. Raw data is provided in Tables 1-2. 
     
       
         
           
               
               
               
               
               
             
               
                   
                 TABLE 1 
               
               
                   
                   
               
               
                   
                   
                 CF20  
                 CF170 
                 CF170/CF20 
               
               
                   
                   
                 (ml/min) 
                 (ml/min) 
                 (ul/ml) 
               
               
                   
                   
               
             
            
               
                   
                 CON11 
                 2.31E+00 
                 1.11E−01 
                 4.81E+01 
               
               
                   
                 CON13 
                 1.07E+00 
                 4.80E−02 
                 4.48E+01 
               
               
                   
                 CON14 
                 8.28E−01 
                 4.50E−02 
                 5.43E+01 
               
               
                   
                 CON9 
                 2.11E+00 
                 6.96E−02 
                 3.30E+01 
               
               
                   
                 CON10 
                 1.85E+00 
                 4.92E−02 
                 2.66E+01 
               
               
                   
                 CON7 
                 1.57E+00 
                 5.40E−02 
                 3.44E+01 
               
               
                   
                 CON8 
                 3.22E+00 
                 6.78E−02 
                 2.11E+01 
               
               
                   
                 CON5 
                 2.18E+00 
                 6.60E−02 
                 3.03E+01 
               
               
                   
                 CON3 
                 2.24E+00 
                 7.92E−02 
                 3.53E+01 
               
               
                   
                 CON4 
                 2.22E+00 
                 7.84E−02 
                 3.53E+01 
               
               
                   
                 CON2 
                 1.68E+00 
                 5.12E−02 
                 3.05E+01 
               
               
                   
                 MEAN 
                 1.93E+00 
                 6.54E−02 
                 3.58E+01 
               
               
                   
                 SD 
                 6.50E−01 
                 1.94E−02 
                 9.72E+00 
               
               
                   
                 SE 
                 1.96E−01 
                 5.86E−03 
                 2.93E+00 
               
               
                   
                 TTEST 
                   
                   
                   
               
               
                   
                 TMZ4 
                 2.13E+00 
                 5.16E−02 
                 2.42E+01 
               
               
                   
                 TMZ3 
                 1.70E+00 
                 1.00E−01 
                 5.87E+01 
               
               
                   
                 TMZ1 
                 2.18E+00 
                 7.78E−02 
                 3.57E+01 
               
               
                   
                 TMZ2 
                 3.83E+00 
                 1.29E−01 
                 3.37E+01 
               
               
                   
                 TMZ7 
                 1.72E+00 
                 8.98E−02 
                 5.21E+01 
               
               
                   
                 TMZ8 
                 2.40E+00 
                 6.56E−02 
                 2.73E+01 
               
               
                   
                 TMZ5 
                 2.14E+00 
                 5.56E−02 
                 2.60E+01 
               
               
                   
                 TMZ9 
                 2.03E+00 
                 1.30E−01 
                 6.39E+01 
               
               
                   
                 MEAN 
                 2.27E+00 
                 8.74E−02 
                 4.02E+01 
               
               
                   
                 SD 
                 6.75E−01 
                 3.06E−02 
                 1.57E+01 
               
               
                   
                 SE 
                 2.39E−01 
                 1.08E−02 
                 5.56E+00 
               
               
                   
                 TTEST 
                   
                   
                   
               
               
                   
                 TNF1 
                 2.24E+00 
                 4.80E−02 
                 2.14E+01 
               
               
                   
                 TNF2 
                 2.24E+00 
                 3.80E−02 
                 1.69E+01 
               
               
                   
                 TNF3 
                 7.32E−01 
                 4.80E−02 
                 6.56E+01 
               
               
                   
                 TNF4 
                 8.20E−01 
                 4.90E−02 
                 5.98E+01 
               
               
                   
                 TNF5 
                 1.09E+00 
                 2.70E−02 
                 2.48E+01 
               
               
                   
                 TNF6 
                 9.48E−01 
                 1.50E−01 
                 1.58E+02 
               
               
                   
                 TNF7 
                 8.08E−01 
                 3.70E−02 
                 4.58E+01 
               
               
                   
                 TNF8 
                 1.20E+00 
                 4.60E−02 
                 3.83E+01 
               
               
                   
                 TNF9 
                 1.45E+00 
                 1.21E−01 
                 8.33E+01 
               
               
                   
                 TNF10 
                 1.20E+00 
                 1.52E−02 
                 1.27E+01 
               
               
                   
                 MEAN 
                 1.27E+00 
                 5.79E−02 
                 5.27E+01 
               
               
                   
                 SD 
                 5.56E−01 
                 4.28E−02 
                 4.37E+01 
               
               
                   
                 SE 
                 1.76E−01 
                 1.35E−02 
                 1.38E+01 
               
               
                   
                 TTEST 
                 2.21E−02 
                 6.06E−01 
                 2.26E−01 
               
               
                   
                 TNS1 
                 1.52E+00 
                 4.70E−02 
                 3.08E+01 
               
               
                   
                 TNS2 
                 9.30E−01 
                 2.90E−02 
                 3.12E+01 
               
               
                   
                 TNS3 
                 2.24E+00 
                 1.67E−01 
                 7.46E+01 
               
               
                   
                 TNS5 
                 5.64E−01 
                 5.00E−02 
                 8.87E+01 
               
               
                   
                 TNS6 
                 6.28E−01 
                 4.40E−02 
                 7.01E+01 
               
               
                   
                 TNS7 
                 1.08E+00 
                 6.40E−02 
                 5.95E+01 
               
               
                   
                 TNS8 
                 8.72E−01 
                 2.30E−02 
                 2.64E+01 
               
               
                   
                 TNS9 
                 1.18E+00 
                 8.50E−02 
                 7.23E+01 
               
               
                   
                 TNS10 
                 1.70E+00 
                 1.84E−01 
                 1.08E+02 
               
               
                   
                 MEAN 
                 1.19E+00 
                 7.70E−02 
                 6.24E+01 
               
               
                   
                 SD 
                 5.43E−01 
                 5.89E−02 
                 2.82E+01 
               
               
                   
                 SE 
                 1.81E−01 
                 1.96E−02 
                 9.42E+00 
               
               
                   
                 TTEST 
                 1.35E−02 
                 5.45E−01 
                 8.80E−03 
               
               
                   
                 vs TMZ 
                   
                   
                 6.82E−02 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
             
               
                 TABLE 2 
               
               
                   
               
               
                   
                 CON 
                 TMZ 
                 TNF 
                 TNS 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 MEAN 
                 36 
                 40 
                 53 
                 62 
               
               
                 SD 
                 10 
                 16 
                 44 
                 28 
               
               
                 SE 
                 3 
                 6 
                 14 
                 9 
               
               
                   
               
            
           
         
       
     
       FIG. 40  is graph of left ventricular developed pressure (LVDP) after IR. Blue bars indicate LVDP at 20 minutes, and orange bars indicate LVDP at 170 minutes. TMZ, TNS, and TNF treatment prevented a decline in cardiac function after IR. Raw data is provided in Tables 3-6. 
     
       
         
           
               
               
               
               
               
               
               
             
               
                 TABLE 3 
               
               
                   
               
               
                   
                 pre- 
                   
                   
                   
                   
                   
               
               
                   
                 ischemia 
                 LVESP 
                 LVEDP 
                 HR 
                 LVDP 
                 LVDP × HR 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 5-18-CN 
                 CON11 
                 6.61E+01 
                 6.20E+00 
                 3.28E+02 
                 5.99E+01 
                 1.97E+04 
               
               
                   
                 CON12 
                 8.15E+01 
                 3.73E+00 
                 3.56E+02 
                 7.78E+01 
                 2.77E+04 
               
               
                 6-10-CN 
                 CON13 
                 8.00E+01 
                 −3.74E+00 
                 1.37E+02 
                 8.37E+01 
                 1.15E+04 
               
               
                   
                 CON14 
                 7.28E+01 
                 6.12E+00 
                 4.54E+02 
                 6.67E+01 
                 3.03E+04 
               
               
                 5-15-CN 
                 CON9 
                 8.07E+01 
                 5.00E+00 
                 1.42E+02 
                 7.57E+01 
                 1.08E+04 
               
               
                   
                 CON10 
                 4.91E+01 
                 1.15E+00 
                 3.21E+02 
                 4.80E+01 
                 1.54E+04 
               
               
                 5-12-CN 
                 CON7 
                 8.55E+01 
                 6.35E+00 
                 3.05E+02 
                 7.91E+01 
                 2.42E+04 
               
               
                   
                 CON8 
                 5.06E+01 
                 1.68E+00 
                 3.04E+02 
                 4.90E+01 
                 1.49E+04 
               
               
                 5-9-CN 
                 CON5 
                 5.45E+01 
                 5.63E+00 
                 2.75E+02 
                 4.89E+01 
                 1.35E+04 
               
               
                   
                 CON6 
                 6.37E+01 
                 4.31E+00 
                 3.08E+02 
                 5.94E+01 
                 1.83E+04 
               
               
                 5-7-CN 
                 CON3 
                 7.32E+01 
                 2.70E+00 
                 2.40E+02 
                 7.05E+01 
                 1.69E+04 
               
               
                   
                 CON4 
                 4.91E+01 
                 1.65E−01 
                 3.14E+02 
                 4.89E+01 
                 1.54E+04 
               
               
                 5-5-CN 
                 CON1 
                 9.48E+01 
                 7.96E+00 
                 3.04E+02 
                 8.68E+01 
                 2.64E+04 
               
               
                   
                 CON2 
                 4.69E+01 
                 1.64E−01 
                 4.02E+02 
                 4.67E+01 
                 1.88E+04 
               
               
                   
                 MEAN 
                 6.77E+01 
                 3.39E+00 
                 2.99E+02 
                 6.44E+01 
                 1.88E+04 
               
               
                   
                 SD 
                 1.58E+01 
                 3.21E+00 
                 8.52E+01 
                 1.46E+01 
                 6.12E+03 
               
               
                   
                 SE 
                 4.21E+00 
                 8.57E−01 
                 2.28E+01 
                 3.91E+00 
                 1.63E+03 
               
               
                   
                 TTEST 
                   
                   
                   
                 2.42E−04 
                   
               
               
                 5-14-TMZ 
                 TMZ3 
                 7.58E+01 
                 6.53E+00 
                 2.63E+02 
                 6.93E+01 
                 1.83E+04 
               
               
                   
                 TMZ4 
                 8.44E+01 
                 5.43E+00 
                 2.93E+02 
                 7.90E+01 
                 2.31E+04 
               
               
                 5-11-TMZ 
                 TMZ1 
                 7.15E+01 
                 6.76E+00 
                 1.66E+02 
                 6.48E+01 
                 1.08E+04 
               
               
                   
                 TMZ2 
                 5.47E+01 
                 1.74E+00 
                 3.35E+02 
                 5.30E+01 
                 1.77E+04 
               
               
                 5-8-TMZ 
                 TMZ7 
                 6.87E+01 
                 3.58E+00 
                 3.58E+02 
                 6.51E+01 
                 2.33E+04 
               
               
                   
                 TMZ8 
                 4.27E+01 
                 4.71E+00 
                 3.33E+02 
                 3.80E+01 
                 1.26E+04 
               
               
                 5-6-TMZ 
                 TMZ5 
                 3.30E+01 
                 4.77E+00 
                 3.48E+02 
                 2.82E+01 
                 9.82E+03 
               
               
                   
                 TMZ6 
                 3.30E+01 
                 1.46E+00 
                 3.21E+02 
                 3.15E+01 
                 1.01E+04 
               
               
                 5-4-TMZ 
                 TMZ9 
                 6.60E+01 
                 7.25E+00 
                 2.67E+02 
                 5.87E+01 
                 1.57E+04 
               
               
                   
                 TMZ10 
                 7.38E+01 
                 2.70E+00 
                 3.32E+02 
                 7.11E+01 
                 2.36E+04 
               
               
                   
                 MEAN 
                 6.03E+01 
                 4.49E+00 
                 3.02E+02 
                 5.59E+01 
                 1.68E+04 
               
               
                   
                 SD 
                 1.85E+01 
                 2.07E+00 
                 5.75E+01 
                 1.77E+01 
                 5.56E+03 
               
               
                   
                 SE 
                 5.84E+00 
                 6.56E−01 
                 1.82E+01 
                 5.58E+00 
                 1.76E+03 
               
               
                   
                   
                   
                   
                   
                 3.85E−01 
                   
               
               
                 5-19-TNF 
                 TNF1 
                 5.02E+01 
                 3.04E+00 
                 4.09E+02 
                 4.72E+01 
                 1.93E+04 
               
               
                   
                 TNF2 
                 4.65E+01 
                 1.76E−01 
                 2.76E+02 
                 4.63E+01 
                 1.28E+04 
               
               
                 6-8-TNF 
                 TNF3 
                 7.13E+01 
                 1.53E+00 
                 6.48E+01 
                 6.97E+01 
                 4.52E+03 
               
               
                   
                 TNF4 
                 9.97E+01 
                 4.15E+00 
                 1.54E+02 
                 9.55E+01 
                 1.47E+04 
               
               
                 6-12-TNF 
                 TNF5 
                 7.14E+01 
                 −3.42E+00 
                 2.77E+02 
                 7.49E+01 
                 2.07E+04 
               
               
                   
                 TNF6 
                 8.98E+01 
                 8.85E+00 
                 3.10E+02 
                 8.09E+01 
                 2.51E+04 
               
               
                 6-14-TNF 
                 TNF7 
                 6.58E+01 
                 7.01E+00 
                 3.98E+02 
                 5.88E+01 
                 2.34E+04 
               
               
                   
                 TNF8 
                 5.99E+01 
                 1.02E+00 
                 2.28E+02 
                 5.89E+01 
                 1.34E+04 
               
               
                 6-15-TNF 
                 TNF9 
                 7.89E+01 
                 2.37E−01 
                 2.71E+02 
                 7.87E+01 
                 2.13E+04 
               
               
                   
                 TNF10 
                 4.01E+01 
                 1.88E+00 
                 3.14E+02 
                 3.82E+01 
                 1.20E+04 
               
               
                   
                 MEAN 
                 6.74E+01 
                 2.45E+00 
                 2.70E+02 
                 6.49E+01 
                 1.67E+04 
               
               
                   
                 SD 
                 1.90E+01 
                 3.54E+00 
                 1.04E+02 
                 1.81E+01 
                 6.32E+03 
               
               
                   
                 SE 
                 6.00E+00 
                 1.12E+00 
                 3.28E+01 
                 5.73E+00 
                 2.00E+03 
               
               
                   
                   
                   
                   
                   
                 1.38E−01 
                   
               
               
                 5-20-TNS 
                 TNS1 
                 5.59E+01 
                 5.23E+00 
                 3.33E+02 
                 5.07E+01 
                 1.69E+04 
               
               
                   
                 TNS2 
                 5.54E+01 
                 −1.83E+00 
                 1.24E+02 
                 5.72E+01 
                 7.09E+03 
               
               
                 6-7-TNS 
                 TNS3 
                 8.78E+01 
                 1.53E+00 
                 1.64E+02 
                 8.63E+01 
                 1.42E+04 
               
               
                   
                 TNS4 
                 1.07E+02 
                 9.86E+00 
                 2.41E+02 
                 9.74E+01 
                 2.35E+04 
               
               
                 6-9-TNS 
                 TNS5 
                 8.97E+01 
                 2.34E+00 
                 8.35E+01 
                 8.74E+01 
                 7.29E+03 
               
               
                   
                 TNS6 
                 6.17E+01 
                 6.21E+00 
                 1.85E+02 
                 5.55E+01 
                 1.03E+04 
               
               
                 6-13-TNS 
                 TNS7 
                 6.62E+01 
                 4.14E+00 
                 3.36E+02 
                 6.21E+01 
                 2.09E+04 
               
               
                   
                 TNS8 
                 6.54E+01 
                 1.22E+01 
                 1.22E+02 
                 5.32E+01 
                 6.47E+03 
               
               
                 6-15-TNS 
                 TNS9 
                 6.16E+01 
                 3.64E+00 
                 3.45E+02 
                 5.80E+01 
                 2.00E+04 
               
               
                   
                 TNS10 
                 5.44E+01 
                 2.47E+00 
                 4.12E+02 
                 5.20E+01 
                 2.14E+04 
               
               
                   
                 MEAN 
                 7.05E+01 
                 4.58E+00 
                 2.35E+02 
                 6.60E+01 
                 1.48E+04 
               
               
                   
                 SD 
                 1.80E+01 
                 4.09E+00 
                 1.15E+02 
                 1.74E+01 
                 6.61E+03 
               
               
                   
                 SE 
                 5.69E+00 
                 1.29E+00 
                 3.63E+01 
                 5.49E+00 
                 2.09E+03 
               
               
                   
                   
                   
                   
                   
                 7.89E−02 
                   
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
               
               
             
               
                 TABLE 4 
               
               
                   
               
               
                   
                 after 2 h 
                   
                   
                   
                   
                   
               
               
                   
                 reperfusion 
                 LVESP 
                 LVEDP 
                 HR 
                 LVDP 
                 LVDP × HR 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 5-18-CN 
                 CON11 
                 7.78E+01 
                 3.68E+01 
                 1.18E+02 
                 4.10E+01 
                 4.82E+03 
               
               
                   
                 CON12 
                 7.07E+01 
                 2.23E+01 
                 9.23E+01 
                 4.84E+01 
                 4.47E+03 
               
               
                 6-10-CN 
                 CON13 
                 6.48E+01 
                 5.54E+01 
                 5.72E+02 
                 9.39E+00 
                 5.38E+03 
               
               
                   
                 CON14 
                 9.54E+01 
                 5.64E+01 
                 2.08E+02 
                 3.90E+01 
                 8.12E+03 
               
               
                 5-15-CN 
                 CON9 
                 5.18E+01 
                 2.71E+01 
                 1.75E+02 
                 2.47E+01 
                 4.33E+03 
               
               
                   
                 CON10 
                 1.10E+02 
                 3.13E+01 
                 5.76E+01 
                 7.84E+01 
                 4.51E+03 
               
               
                 5-12-CN 
                 CON7 
                 3.93E+01 
                 1.42E+01 
                 9.11E+01 
                 2.51E+01 
                 2.29E+03 
               
               
                   
                 CON8 
                 5.29E+01 
                 9.48E+00 
                 6.07E+01 
                 4.34E+01 
                 2.64E+03 
               
               
                 5-9-CN 
                 CON5 
                 6.56E+01 
                 4.89E+01 
                 6.50E+01 
                 1.67E+01 
                 1.09E+03 
               
               
                   
                 CON6 
                 7.44E+01 
                 6.56E+01 
                 3.78E+01 
                 8.81E+00 
                 3.33E+02 
               
               
                 5-7-CN 
                 CON3 
                 6.35E+01 
                 9.99E+00 
                 1.15E+02 
                 5.35E+01 
                 6.18E+03 
               
               
                   
                 CON4 
                 8.76E+01 
                 5.34E+01 
                 1.06E+02 
                 3.43E+01 
                 3.65E+03 
               
               
                 5-5-CN 
                 CON1 
                 9.29E+01 
                 4.38E+01 
                 2.61E+02 
                 4.91E+01 
                 1.28E+04 
               
               
                   
                 CON2 
                 5.18E+01 
                 4.43E+00 
                 2.57E+02 
                 4.74E+01 
                 1.22E+04 
               
               
                   
                 MEAN 
                 7.13E+01 
                 3.42E+01 
                 1.58E+02 
                 3.71E+01 
                 5.20E+03 
               
               
                   
                 SD 
                 1.98E+01 
                 2.02E+01 
                 1.39E+02 
                 1.90E+01 
                 3.68E+03 
               
               
                   
                 SE 
                 5.29E+00 
                 5.40E+00 
                 3.72E+01 
                 5.08E+00 
                 9.83E+02 
               
               
                   
                 TTEST 
                   
                   
                   
                   
                   
               
               
                 5-14-TMZ 
                 TMZ3 
                 5.07E+01 
                 2.93E+01 
                 1.18E+02 
                 2.14E+01 
                 2.52E+03 
               
               
                   
                 TMZ4 
                 7.66E+01 
                 3.31E+01 
                 1.19E+02 
                 4.34E+01 
                 5.15E+03 
               
               
                 5-11-TMZ 
                 TMZ1 
                 9.19E+01 
                 3.96E+01 
                 1.01E+02 
                 5.22E+01 
                 5.28E+03 
               
               
                   
                 TMZ2 
                 4.77E+01 
                 1.80E+01 
                 1.51E+02 
                 2.97E+01 
                 4.49E+03 
               
               
                 5-8-TMZ 
                 TMZ7 
                 5.18E+01 
                 3.36E+00 
                 6.70E+01 
                 4.84E+01 
                 3.24E+03 
               
               
                   
                 TMZ8 
                 4.86E+01 
                 1.87E+00 
                 9.22E+01 
                 4.67E+01 
                 4.31E+03 
               
               
                 5-6-TMZ 
                 TMZ5 
                 6.09E+01 
                 1.99E+01 
                 2.22E+02 
                 4.10E+01 
                 9.11E+03 
               
               
                   
                 TMZ6 
                 1.09E+02 
                 3.21E+01 
                 1.70E+02 
                 7.65E+01 
                 1.30E+04 
               
               
                 5-4-TMZ 
                 TMZ9 
                 7.38E+01 
                 1.84E+01 
                 1.16E+02 
                 5.53E+01 
                 6.44E+03 
               
               
                   
                 TMZ10 
                 7.61E+01 
                 1.77E+00 
                 2.38E+02 
                 7.43E+01 
                 1.77E+04 
               
               
                   
                 MEAN 
                 6.86E+01 
                 1.97E+01 
                 1.39E+02 
                 4.89E+01 
                 6.82E+03 
               
               
                   
                 SD 
                 2.05E+01 
                 1.39E+01 
                 5.58E+01 
                 1.73E+01 
                 4.82E+03 
               
               
                   
                 SE 
                 6.49E+00 
                 4.39E+00 
                 1.77E+01 
                 5.46E+00 
                 1.52E+03 
               
               
                 5-19-TNF 
                 TNF1 
                 8.37E+01 
                 6.66E+01 
                 1.53E+02 
                 1.71E+01 
                 2.62E+03 
               
               
                   
                 TNF2 
                 6.19E+00 
                 5.54E+00 
                 2.13E+03 
                 6.48E−01 
                 1.38E+03 
               
               
                 6-8-TNF 
                 TNF3 
                 8.99E+01 
                 1.88E+01 
                 1.05E+01 
                 7.11E+01 
                 7.49E+02 
               
               
                   
                 TNF4 
                 6.06E+01 
                 1.34E+01 
                 8.10E+01 
                 4.72E+01 
                 3.82E+03 
               
               
                 6-12-TNF 
                 TNF5 
                 1.54E+02 
                 4.15E+01 
                 2.20E+01 
                 1.13E+02 
                 2.48E+03 
               
               
                   
                 TNF6 
                 1.30E+02 
                 4.25E+01 
                 3.33E+01 
                 8.77E+01 
                 2.92E+03 
               
               
                 6-14-TNF 
                 TNF7 
                 5.70E+01 
                 4.00E+01 
                 4.00E+01 
                 1.70E+01 
                 6.80E+02 
               
               
                   
                 TNF8 
                 3.76E+01 
                 1.87E+01 
                 5.36E+01 
                 1.88E+01 
                 1.01E+03 
               
               
                 6-15-TNF 
                 TNF9 
                 6.23E+01 
                 3.38E+01 
                 1.97E+02 
                 2.85E+01 
                 5.59E+03 
               
               
                   
                 TNF10 
                 7.85E+01 
                 2.75E+01 
                 7.85E+01 
                 5.10E+01 
                 4.00E+03 
               
               
                   
                 MEAN 
                 7.60E+01 
                 3.09E+01 
                 2.80E+02 
                 4.52E+01 
                 2.53E+03 
               
               
                   
                 SD 
                 4.28E+01 
                 1.79E+01 
                 6.54E+02 
                 3.59E+01 
                 1.62E+03 
               
               
                   
                 SE 
                 1.35E+01 
                 5.65E+00 
                 2.07E+02 
                 1.14E+01 
                 5.12E+02 
               
               
                 5-20-TNS 
                 TNS1 
                 6.47E+01 
                 1.78E+01 
                 1.04E+02 
                 4.69E+01 
                 4.88E+03 
               
               
                   
                 TNS2 
                 8.95E+01 
                 3.03E+01 
                 5.55E+01 
                 5.92E+01 
                 3.29E+03 
               
               
                 6-7-TNS 
                 TNS3 
                 7.79E+01 
                 6.34E+01 
                 1.28E+02 
                 1.45E+01 
                 1.85E+03 
               
               
                   
                 TNS4 
                 7.74E+01 
                 2.73E+01 
                 1.02E+02 
                 5.01E+01 
                 5.09E+03 
               
               
                 6-9-TNS 
                 TNS5 
                 1.37E+02 
                 5.63E+01 
                 1.63E+01 
                 8.08E+01 
                 1.32E+03 
               
               
                   
                 TNS6 
                 8.59E+01 
                 1.23E+01 
                 1.06E+02 
                 7.36E+01 
                 7.79E+03 
               
               
                 6-13-TNS 
                 TNS7 
                 5.76E+01 
                 5.16E+01 
                 1.35E+02 
                 6.00E+00 
                 8.07E+02 
               
               
                   
                 TNS8 
                 4.96E+01 
                 1.53E+01 
                 1.22E+02 
                 3.43E+01 
                 4.20E+03 
               
               
                 6-15-TNS 
                 TNS9 
                 9.97E+01 
                 3.00E+01 
                 7.46E+01 
                 6.98E+01 
                 5.21E+03 
               
               
                   
                 TNS10 
                 4.32E+01 
                 −4.32E+00 
                 7.20E+01 
                 4.75E+01 
                 3.42E+03 
               
               
                   
                 MEAN 
                 7.83E+01 
                 3.00E+01 
                 9.15E+01 
                 4.83E+01 
                 3.79E+03 
               
               
                   
                 SD 
                 2.74E+01 
                 2.14E+01 
                 3.69E+01 
                 2.45E+01 
                 2.11E+03 
               
               
                   
                 SE 
                 8.67E+00 
                 6.78E+00 
                 1.17E+01 
                 7.75E+00 
                 6.69E+02 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
               
               
             
               
                 TABLE 5 
               
               
                   
               
               
                   
                 pre- 
                   
                   
                 after 2 h 
                   
                   
               
               
                   
                 ischemia 
                 +dp/dtm 
                 −dp/dtm 
                 reperfusion 
                 +dp/dtm 
                 −dp/dtm 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 5-18-CN 
                 CON11 
                 2.60E+03 
                 −1.82E+03 
                 CON11 
                 1.44E+03 
                 −8.67E+02 
               
               
                   
                 CON12 
                 2.95E+03 
                 −2.58E+03 
                 CON12 
                 1.63E+03 
                 −1.07E+03 
               
               
                 6-10-CN 
                 CON13 
                 3.10E+03 
                 −2.42E+03 
                 CON13 
                 2.25E+02 
                 −2.22E+02 
               
               
                   
                 CON14 
                 3.08E+03 
                 −2.10E+03 
                 CON14 
                 3.44E+02 
                 −2.87E+02 
               
               
                 5-15-CN 
                 CON9 
                 2.28E+03 
                 −1.38E+03 
                 CON9 
                 9.45E+02 
                 −5.54E+02 
               
               
                   
                 CON10 
                 2.06E+03 
                 −1.50E+03 
                 CON10 
                 2.29E+03 
                 −1.75E+03 
               
               
                 5-12-CN 
                 CON7 
                 2.71E+03 
                 −2.10E+03 
                 CON7 
                 2.51E+02 
                 −2.55E+02 
               
               
                   
                 CON8 
                 1.58E+03 
                 −1.10E+03 
                 CON8 
                 3.63E+02 
                 −3.05E+02 
               
               
                 5-9-CN 
                 CON5 
                 2.17E+03 
                 −1.50E+03 
                 CON5 
                 2.39E+02 
                 −2.41E+02 
               
               
                   
                 CON6 
                 2.25E+03 
                 −1.62E+03 
                 CON6 
                 1.47E+02 
                 −1.49E+02 
               
               
                 5-7-CN 
                 CON3 
                 2.63E+03 
                 −2.06E+03 
                 CON3 
                 1.63E+03 
                 −1.06E+03 
               
               
                   
                 CON4 
                 2.05E+03 
                 −1.38E+03 
                 CON4 
                 1.10E+03 
                 −7.03E+02 
               
               
                 5-5-CN 
                 CON1 
                 3.17E+03 
                 −2.37E+03 
                 CON1 
                 1.03E+03 
                 −1.12E+03 
               
               
                   
                 CON2 
                 2.10E+03 
                 −1.50E+03 
                 CON2 
                 1.75E+03 
                 −1.27E+03 
               
               
                   
                 MEAN 
                 2.48E+03 
                 −1.82E+03 
                 MEAN 
                 9.56E+02 
                 −7.04E+02 
               
               
                   
                 SD 
                 4.84E+02 
                 4.56E+02 
                 SD 
                 7.08E+02 
                 4.95E+02 
               
               
                   
                 SE 
                 1.29E+02 
                 1.22E+02 
                 SE 
                 1.89E+02 
                 1.32E+02 
               
               
                   
                 TTEST 
                   
                   
                 TTEST 
                   
                   
               
               
                 5-14-TMZ 
                 TMZ3 
                 2.41E+03 
                 −1.69E+03 
                 TMZ3 
                 4.14E+02 
                 −3.57E+02 
               
               
                   
                 TMZ4 
                 2.77E+03 
                 −2.26E+03 
                 TMZ4 
                 1.48E+03 
                 −1.15E+03 
               
               
                 5-11-TMZ 
                 TMZ1 
                 1.80E+03 
                 −1.59E+03 
                 TMZ1 
                 1.38E+03 
                 −7.45E+02 
               
               
                   
                 TMZ2 
                 2.15E+03 
                 −1.80E+03 
                 TMZ2 
                 1.06E+03 
                 −6.85E+02 
               
               
                 5-8-TMZ 
                 TMZ7 
                 3.40E+03 
                 −2.59E+03 
                 TMZ7 
                 3.44E+02 
                 −3.39E+02 
               
               
                   
                 TMZ8 
                 1.75E+03 
                 −1.20E+03 
                 TMZ8 
                 7.36E+02 
                 −4.28E+02 
               
               
                 5-6-TMZ 
                 TMZ5 
                 1.27E+03 
                 −8.82E+02 
                 TMZ5 
                 1.28E+03 
                 −8.38E+02 
               
               
                   
                 TMZ6 
                 1.24E+03 
                 −6.59E+02 
                 TMZ6 
                 1.85E+03 
                 −1.06E+03 
               
               
                 5-4-TMZ 
                 TMZ9 
                 1.98E+03 
                 −1.41E+03 
                 TMZ9 
                 1.13E+03 
                 −6.38E+02 
               
               
                   
                 TMZ10 
                 2.02E+03 
                 −1.56E+03 
                 TMZ10 
                 1.62E+03 
                 −9.83E+02 
               
               
                   
                 MEAN 
                 2.08E+03 
                 −1.56E+03 
                 MEAN 
                 1.13E+03 
                 −7.22E+02 
               
               
                   
                 SD 
                 6.58E+02 
                 5.81E+02 
                 SD 
                 5.01E+02 
                 2.90E+02 
               
               
                   
                 SE 
                 2.08E+02 
                 1.84E+02 
                 SE 
                 1.58E+02 
                 9.16E+01 
               
               
                   
                   
                   
                   
                   
                 5.16E−01 
                 9.18E−01 
               
               
                 5-19-TNF 
                 TNF1 
                 2.67E+03 
                 −1.49E+03 
                 TNF1 
                 3.86E+02 
                 −3.75E+02 
               
               
                   
                 TNF2 
                 2.85E+03 
                 −1.44E+03 
                 TNF2 
                 1.46E+02 
                 −1.43E+02 
               
               
                 6-8-TNF 
                 TNF3 
                 1.53E+03 
                 −7.24E+02 
                 TNF3 
                 2.28E+02 
                 −2.34E+02 
               
               
                   
                 TNF4 
                 3.86E+03 
                 −2.59E+03 
                 TNF4 
                 2.84E+02 
                 −2.40E+02 
               
               
                 6-12-TNF 
                 TNF5 
                 3.29E+03 
                 −2.34E+03 
                 TNF5 
                 2.92E+03 
                 −2.08E+03 
               
               
                   
                 TNF6 
                 3.03E+03 
                 −1.90E+03 
                 TNF6 
                 2.48E+03 
                 −1.84E+03 
               
               
                 6-14-TNF 
                 TNF7 
                 3.22E+03 
                 −1.62E+03 
                 TNF7 
                 2.53E+02 
                 −2.48E+02 
               
               
                   
                 TNF8 
                 1.74E+03 
                 −1.12E+03 
                 TNF8 
                 1.53E+02 
                 −1.52E+02 
               
               
                 6-15-TNF 
                 TNF9 
                 2.14E+03 
                 −2.33E+03 
                 TNF9 
                 1.04E+03 
                 −6.31E+02 
               
               
                   
                 TNF10 
                 1.86E+03 
                 −9.97E+02 
                 TNF10 
                 2.04E+03 
                 −1.34E+03 
               
               
                   
                 MEAN 
                 2.62E+03 
                 −1.65E+03 
                 MEAN 
                 9.93E+02 
                 −7.29E+02 
               
               
                   
                 SD 
                 7.71E+02 
                 6.26E+02 
                 SD 
                 1.08E+03 
                 7.43E+02 
               
               
                   
                 SE 
                 2.44E+02 
                 1.98E+02 
                 SE 
                 3.41E+02 
                 2.35E+02 
               
               
                   
                   
                   
                   
                   
                 1.09E−03 
                 7.48E−03 
               
               
                 5-20-TNS 
                 TNS1 
                 2.37E+03 
                 −1.60E+03 
                 TNS1 
                 1.79E+03 
                 −1.12E+03 
               
               
                   
                 TNS2 
                 2.87E+03 
                 −2.53E+03 
                 TNS2 
                 1.84E+03 
                 −1.30E+03 
               
               
                 6-7-TNS 
                 TNS3 
                 4.00E+03 
                 −2.67E+03 
                 TNS3 
                 2.91E+02 
                 −3.02E+02 
               
               
                   
                 TNS4 
                 3.32E+03 
                 −2.63E+03 
                 TNS4 
                 1.62E+03 
                 −1.30E+03 
               
               
                 6-9-TNS 
                 TNS5 
                 3.36E+03 
                 −2.21E+03 
                 TNS5 
                 2.43E+02 
                 −2.46E+02 
               
               
                   
                 TNS6 
                 2.53E+03 
                 −1.89E+03 
                 TNS6 
                 2.36E+03 
                 −1.74E+03 
               
               
                 6-13-TNS 
                 TNS7 
                 2.92E+03 
                 −1.75E+03 
                 TNS7 
                 2.49E+02 
                 −2.47E+02 
               
               
                   
                 TNS8 
                 1.12E+03 
                 −7.42E+02 
                 TNS8 
                 1.29E+03 
                 −8.50E+02 
               
               
                 6-15-TNS 
                 TNS9 
                 2.29E+03 
                 −1.75E+03 
                 TNS9 
                 2.06E+03 
                 −1.59E+03 
               
               
                   
                 TNS10 
                 2.11E+03 
                 −1.58E+03 
                 TNS10 
                 1.26E+03 
                 −1.10E+03 
               
               
                   
                 MEAN 
                 2.69E+03 
                 −1.94E+03 
                 MEAN 
                 1.30E+03 
                 −9.80E+02 
               
               
                   
                 SD 
                 7.99E+02 
                 5.95E+02 
                 SD 
                 7.86E+02 
                 5.52E+02 
               
               
                   
                 SE 
                 2.53E+02 
                 1.88E+02 
                 SE 
                 2.49E+02 
                 1.75E+02 
               
               
                   
                   
                 9.96E−04 
                 1.55E−03 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
               
               
               
             
               
                   
                 TABLE 6 
               
               
                   
                   
               
               
                   
                   
                   
                 CON 
                 TMZ 
                 TNF 
                 TNS 
               
               
                   
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                 T20 
                 Mean 
                 64.36 
                 55.86 
                 64.90 
                 65.96 
               
               
                   
                 T20 
                 SE 
                 3.91 
                 5.58 
                 5.73 
                 5.49 
               
               
                   
                 T170 
                 Mean 
                 37.09 
                 48.91 
                 45.16 
                 48.27 
               
               
                   
                 T170 
                 SE 
                 5.08 
                 5.46 
                 11.36 
                 7.75 
               
               
                   
                   
               
            
           
         
       
     
       FIG. 41  shows images of TTC-stained heart slices after IR. TMZ and TNS treatment decreased infarct size after IR. 
       FIG. 42  is graph of infarct size after IR. TMZ and TNS treatment decreased infarct size after IR. Raw data is provided in Tables 7-55. 
     
       
         
           
               
             
               
                 TABLE 7 
               
               
                   
               
               
                 CN11 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide11.jpg 
                 1649 
                   
               
               
                   
                  2 
                 Slide11.jpg 
                 10 
                 0.06 
               
               
                   
                  3 
                 Slide11.jpg 
                 1385 
                 8.40 
               
               
                   
                  4 
                 Slide11.jpg 
                 2808 
                   
               
               
                   
                  5 
                 Slide11.jpg 
                 104 
                 0.81 
               
               
                   
                  6 
                 Slide11.jpg 
                 2525 
                 19.78 
               
               
                   
                  7 
                 Slide11.jpg 
                 3807 
                   
               
               
                   
                  8 
                 Slide11.jpg 
                 1014 
                 7.99 
               
               
                   
                  9 
                 Slide11.jpg 
                 2207 
                 17.39 
               
               
                   
                 10 
                 Slide11.jpg 
                 3952 
                   
               
               
                   
                 11 
                 Slide11.jpg 
                 15 
                 0.08 
               
               
                   
                 12 
                 Slide11.jpg 
                 3300 
                 17.54 
               
               
                   
                 13 
                 Slide11.jpg 
                 3376 
                   
               
               
                   
                 14 
                 Slide11.jpg 
                 103 
                 0.92 
               
               
                   
                 15 
                 Slide11.jpg 
                 2816 
                 25.02 
               
               
                   
                 16 
                 Slide11.jpg 
                 1616 
                   
               
               
                   
                 17 
                 Slide11.jpg 
                 975 
                 6.03 
               
               
                   
                 18 
                 Slide11.jpg 
                 409 
                 2.53 
               
               
                   
                 19 
                 Slide11.jpg 
                 2805 
                   
               
               
                   
                 20 
                 Slide11.jpg 
                 819 
                 6.42 
               
               
                   
                 21 
                 Slide11.jpg 
                 1496 
                 11.73 
               
               
                   
                 22 
                 Slide11.jpg 
                 3973 
                   
               
               
                   
                 23 
                 Slide11.jpg 
                 1047 
                 7.91 
               
               
                   
                 24 
                 Slide11.jpg 
                 2465 
                 18.61 
               
               
                   
                 25 
                 Slide11.jpg 
                 3971 
                   
               
               
                   
                 26 
                 Slide11.jpg 
                 1102 
                 5.83 
               
               
                   
                 27 
                 Slide11.jpg 
                 2430 
                 12.85 
               
               
                   
                 28 
                 Slide11.jpg 
                 3516 
                   
               
               
                   
                 29 
                 Slide11.jpg 
                 1919 
                 16.37 
               
               
                   
                 30 
                 Slide11.jpg 
                 920 
                 7.85 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
               
                   
               
               
                 CN11 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 26.21 
               
               
                   
                 IS 
                 70.86 
               
               
                   
                 LV 
                 97.07 
               
               
                   
                 IS/LV 
                 73% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
               
                   
               
               
                 CN12 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide12.jpg 
                 1562 
                   
               
               
                   
                  2 
                 Slide12.jpg 
                 1059 
                 8.81 
               
               
                   
                  3 
                 Slide12.jpg 
                 485 
                 4.04 
               
               
                   
                  4 
                 Slide12.jpg 
                 2925 
                   
               
               
                   
                  5 
                 Slide12.jpg 
                 260 
                 1.78 
               
               
                   
                  6 
                 Slide12.jpg 
                 2159 
                 14.76 
               
               
                   
                  7 
                 Slide12.jpg 
                 3492 
                   
               
               
                   
                  8 
                 Slide12.jpg 
                 263 
                 1.88 
               
               
                   
                  9 
                 Slide12.jpg 
                 2886 
                 20.66 
               
               
                   
                 10 
                 Slide12.jpg 
                 4855 
                   
               
               
                   
                 11 
                 Slide12.jpg 
                 1992 
                 16.00 
               
               
                   
                 12 
                 Slide12.jpg 
                 2292 
                 18.41 
               
               
                   
                 13 
                 Slide12.jpg 
                 2934 
                   
               
               
                   
                 14 
                 Slide12.jpg 
                 1405 
                 6.70 
               
               
                   
                 15 
                 Slide12.jpg 
                 914 
                 4.36 
               
               
                   
                 16 
                 Slide12.jpg 
                 2061 
                   
               
               
                   
                 17 
                 Slide12.jpg 
                 81 
                 0.51 
               
               
                   
                 18 
                 Slide12.jpg 
                 1704 
                 10.75 
               
               
                   
                 19 
                 Slide12.jpg 
                 2966 
                   
               
               
                   
                 20 
                 Slide12.jpg 
                 105 
                 0.71 
               
               
                   
                 21 
                 Slide12.jpg 
                 2810 
                 18.95 
               
               
                   
                 22 
                 Slide12.jpg 
                 4099 
                   
               
               
                   
                 23 
                 Slide12.jpg 
                 823 
                 5.02 
               
               
                   
                 24 
                 Slide12.jpg 
                 2350 
                 14.33 
               
               
                   
                 25 
                 Slide12.jpg 
                 3979 
                   
               
               
                   
                 26 
                 Slide12.jpg 
                 357 
                 3.50 
               
               
                   
                 27 
                 Slide12.jpg 
                 2787 
                 27.32 
               
               
                   
                 28 
                 Slide12.jpg 
                 2974 
                   
               
               
                   
                 29 
                 Slide12.jpg 
                 490 
                 2.31 
               
               
                   
                 30 
                 Slide12.jpg 
                 2112 
                 9.94 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
               
                   
               
               
                 CN12 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 23.61 
               
               
                   
                 IS 
                 71.76 
               
               
                   
                 LV 
                 95.37 
               
               
                   
                 IS/LV 
                 75% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
               
                   
               
               
                 TNS1 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide15.jpg 
                 1857 
                   
               
               
                   
                  2 
                 Slide15.jpg 
                 58 
                 0.28 
               
               
                   
                  3 
                 Slide15.jpg 
                 1672 
                 8.10 
               
               
                   
                  4 
                 Slide15.jpg 
                 3383 
                   
               
               
                   
                  5 
                 Slide15.jpg 
                 901 
                 4.53 
               
               
                   
                  6 
                 Slide15.jpg 
                 1873 
                 9.41 
               
               
                   
                  7 
                 Slide15.jpg 
                 3460 
                   
               
               
                   
                  8 
                 Slide15.jpg 
                 1452 
                 13.43 
               
               
                   
                  9 
                 Slide15.jpg 
                 2272 
                 21.01 
               
               
                   
                 10 
                 Slide15.jpg 
                 3712 
                   
               
               
                   
                 11 
                 Slide15.jpg 
                 772 
                 8.32 
               
               
                   
                 12 
                 Slide15.jpg 
                 2422 
                 26.10 
               
               
                   
                 13 
                 Slide15.jpg 
                 3088 
                   
               
               
                   
                 14 
                 Slide15.jpg 
                 498 
                 3.87 
               
               
                   
                 15 
                 Slide15.jpg 
                 1733 
                 13.47 
               
               
                   
                 16 
                 Slide15.jpg 
                 1762 
                   
               
               
                   
                 17 
                 Slide15.jpg 
                 65 
                 0.33 
               
               
                   
                 18 
                 Slide15.jpg 
                 1626 
                 8.31 
               
               
                   
                 19 
                 Slide15.jpg 
                 3532 
                   
               
               
                   
                 20 
                 Slide15.jpg 
                 2034 
                 9.79 
               
               
                   
                 21 
                 Slide15.jpg 
                 1206 
                 5.80 
               
               
                   
                 22 
                 Slide15.jpg 
                 3411 
                   
               
               
                   
                 23 
                 Slide15.jpg 
                 1752 
                 16.44 
               
               
                   
                 24 
                 Slide15.jpg 
                 1006 
                 9.44 
               
               
                   
                 25 
                 Slide15.jpg 
                 4241 
                   
               
               
                   
                 26 
                 Slide15.jpg 
                 2148 
                 20.26 
               
               
                   
                 27 
                 Slide15.jpg 
                 1101 
                 10.38 
               
               
                   
                 28 
                 Slide15.jpg 
                 3440 
                   
               
               
                   
                 29 
                 Slide15.jpg 
                 2307 
                 16.10 
               
               
                   
                 30 
                 Slide15.jpg 
                 165 
                 1.15 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 12 
               
               
                   
               
               
                 TNS1 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 46.67 
               
               
                   
                 IS 
                 56.59 
               
               
                   
                 LV 
                 103.26 
               
               
                   
                 IS/LV 
                 55% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 13 
               
               
                   
               
               
                 TNS2 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide16.jpg 
                 1565 
                   
               
               
                   
                  2 
                 Slide16.jpg 
                 1058 
                 7.44 
               
               
                   
                  3 
                 Slide16.jpg 
                 145 
                 1.02 
               
               
                   
                  4 
                 Slide16.jpg 
                 2654 
                   
               
               
                   
                  5 
                 Slide16.jpg 
                 431 
                 3.90 
               
               
                   
                  6 
                 Slide16.jpg 
                 2043 
                 18.47 
               
               
                   
                  7 
                 Slide16.jpg 
                 3247 
                   
               
               
                   
                  8 
                 Slide16.jpg 
                 1053 
                 8.43 
               
               
                   
                  9 
                 Slide16.jpg 
                 1584 
                 12.68 
               
               
                   
                 10 
                 Slide16.jpg 
                 3892 
                   
               
               
                   
                 11 
                 Slide16.jpg 
                 2391 
                 22.73 
               
               
                   
                 12 
                 Slide16.jpg 
                 863 
                 8.20 
               
               
                   
                 13 
                 Slide16.jpg 
                 2505 
                   
               
               
                   
                 14 
                 Slide16.jpg 
                 1488 
                 14.85 
               
               
                   
                 15 
                 Slide16.jpg 
                 363 
                 3.62 
               
               
                   
                 16 
                 Slide16.jpg 
                 1526 
                   
               
               
                   
                 17 
                 Slide16.jpg 
                 9 
                 0.06 
               
               
                   
                 18 
                 Slide16.jpg 
                 1357 
                 9.78 
               
               
                   
                 19 
                 Slide16.jpg 
                 2337 
                   
               
               
                   
                 20 
                 Slide16.jpg 
                 16 
                 0.16 
               
               
                   
                 21 
                 Slide16.jpg 
                 1899 
                 19.50 
               
               
                   
                 22 
                 Slide16.jpg 
                 3558 
                   
               
               
                   
                 23 
                 Slide16.jpg 
                 1453 
                 10.62 
               
               
                   
                 24 
                 Slide16.jpg 
                 1504 
                 10.99 
               
               
                   
                 25 
                 Slide16.jpg 
                 4041 
                   
               
               
                   
                 26 
                 Slide16.jpg 
                 517 
                 4.73 
               
               
                   
                 27 
                 Slide16.jpg 
                 2763 
                 25.30 
               
               
                   
                 28 
                 Slide16.jpg 
                 2946 
                   
               
               
                   
                 29 
                 Slide16.jpg 
                 631 
                 5.35 
               
               
                   
                 30 
                 Slide16.jpg 
                 1326 
                 11.25 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 14 
               
               
                   
               
               
                 TNS2 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 39.14 
               
               
                   
                 IS 
                 60.41 
               
               
                   
                 LV 
                 99.56 
               
               
                   
                 IS/LV 
                 61% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 15 
               
               
                   
               
               
                 TNF1 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide17.jpg 
                 1326 
                   
               
               
                   
                  2 
                 Slide17.jpg 
                 63 
                 0.24 
               
               
                   
                  3 
                 Slide17.jpg 
                 1183 
                 4.46 
               
               
                   
                  4 
                 Slide17.jpg 
                 3158 
                   
               
               
                   
                  5 
                 Slide17.jpg 
                 825 
                 5.49 
               
               
                   
                  6 
                 Slide17.jpg 
                 2014 
                 13.39 
               
               
                   
                  7 
                 Slide17.jpg 
                 4805 
                   
               
               
                   
                  8 
                 Slide17.jpg 
                 1774 
                 12.92 
               
               
                   
                  9 
                 Slide17.jpg 
                 1722 
                 12.54 
               
               
                   
                 10 
                 Slide17.jpg 
                 4675 
                   
               
               
                   
                 11 
                 Slide17.jpg 
                 1984 
                 15.28 
               
               
                   
                 12 
                 Slide17.jpg 
                 2470 
                 19.02 
               
               
                   
                 13 
                 Slide17.jpg 
                 2754 
                   
               
               
                   
                 14 
                 Slide17.jpg 
                 269 
                 2.05 
               
               
                   
                 15 
                 Slide17.jpg 
                 1377 
                 10.50 
               
               
                   
                 16 
                 Slide17.jpg 
                 1373 
                   
               
               
                   
                 17 
                 Slide17.jpg 
                 1067 
                 3.89 
               
               
                   
                 18 
                 Slide17.jpg 
                 43 
                 0.16 
               
               
                   
                 19 
                 Slide17.jpg 
                 3113 
                   
               
               
                   
                 20 
                 Slide17.jpg 
                 803 
                 5.42 
               
               
                   
                 21 
                 Slide17.jpg 
                 2008 
                 13.55 
               
               
                   
                 22 
                 Slide17.jpg 
                 4657 
                   
               
               
                   
                 23 
                 Slide17.jpg 
                 1189 
                 8.94 
               
               
                   
                 24 
                 Slide17.jpg 
                 2398 
                 18.02 
               
               
                   
                 25 
                 Slide17.jpg 
                 4607 
                   
               
               
                   
                 26 
                 Slide17.jpg 
                 1256 
                 9.81 
               
               
                   
                 27 
                 Slide17.jpg 
                 1978 
                 15.46 
               
               
                   
                 28 
                 Slide17.jpg 
                 2769 
                   
               
               
                   
                 29 
                 Slide17.jpg 
                 2115 
                 16.04 
               
               
                   
                 30 
                 Slide17.jpg 
                 72 
                 0.55 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 16 
               
               
                   
               
               
                 TNF1 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 40.03 
               
               
                   
                 IS 
                 53.82 
               
               
                   
                 LV 
                 93.86 
               
               
                   
                 IS/LV 
                 57% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 17 
               
               
                   
               
               
                 TNF2 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide18.jpg 
                 2133 
                   
               
               
                   
                  2 
                 Slide18.jpg 
                 1861 
                 12.21 
               
               
                   
                  3 
                 Slide18.jpg 
                 239 
                 1.57 
               
               
                   
                  4 
                 Slide18.jpg 
                 4037 
                   
               
               
                   
                  5 
                 Slide18.jpg 
                 753 
                 5.60 
               
               
                   
                  6 
                 Slide18.jpg 
                 2304 
                 17.12 
               
               
                   
                  7 
                 Slide18.jpg 
                 4663 
                   
               
               
                   
                  8 
                 Slide18.jpg 
                 1548 
                 10.62 
               
               
                   
                  9 
                 Slide18.jpg 
                 2917 
                 20.02 
               
               
                   
                 10 
                 Slide18.jpg 
                 5017 
                   
               
               
                   
                 11 
                 Slide18.jpg 
                 2648 
                 20.06 
               
               
                   
                 12 
                 Slide18.jpg 
                 2480 
                 18.78 
               
               
                   
                 13 
                 Slide18.jpg 
                 3629 
                   
               
               
                   
                 14 
                 Slide18.jpg 
                 1698 
                 13.10 
               
               
                   
                 15 
                 Slide18.jpg 
                 348 
                 2.69 
               
               
                   
                 16 
                 Slide18.jpg 
                 2130 
                   
               
               
                   
                 17 
                 Slide18.jpg 
                 4 
                 0.03 
               
               
                   
                 18 
                 Slide18.jpg 
                 1988 
                 13.07 
               
               
                   
                 19 
                 Slide18.jpg 
                 4108 
                   
               
               
                   
                 20 
                 Slide18.jpg 
                 253 
                 1.85 
               
               
                   
                 21 
                 Slide18.jpg 
                 3796 
                 27.72 
               
               
                   
                 22 
                 Slide18.jpg 
                 4612 
                   
               
               
                   
                 23 
                 Slide18.jpg 
                 815 
                 5.65 
               
               
                   
                 24 
                 Slide18.jpg 
                 2427 
                 16.84 
               
               
                   
                 25 
                 Slide18.jpg 
                 4880 
                   
               
               
                   
                 26 
                 Slide18.jpg 
                 562 
                 4.38 
               
               
                   
                 27 
                 Slide18.jpg 
                 3535 
                 27.53 
               
               
                   
                 28 
                 Slide18.jpg 
                 3507 
                   
               
               
                   
                 29 
                 Slide18.jpg 
                 497 
                 3.97 
               
               
                   
                 30 
                 Slide18.jpg 
                 1837 
                 14.67 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 18 
               
               
                   
               
               
                 TNF2 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 38.73 
               
               
                   
                 IS 
                 80.00 
               
               
                   
                 LV 
                 118.73 
               
               
                   
                 IS/LV 
                 73% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 19 
               
               
                   
               
               
                 TNS3 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide19.jpg 
                 1484 
                   
               
               
                   
                  2 
                 Slide19.jpg 
                 923 
                 4.98 
               
               
                   
                  3 
                 Slide19.jpg 
                 714 
                 3.85 
               
               
                   
                  4 
                 Slide19.jpg 
                 3124 
                   
               
               
                   
                  5 
                 Slide19.jpg 
                 990 
                 6.65 
               
               
                   
                  6 
                 Slide19.jpg 
                 1845 
                 12.40 
               
               
                   
                  7 
                 Slide19.jpg 
                 3414 
                   
               
               
                   
                  8 
                 Slide19.jpg 
                 1282 
                 13.89 
               
               
                   
                  9 
                 Slide19.jpg 
                 1833 
                 19.87 
               
               
                   
                 10 
                 Slide19.jpg 
                 3380 
                   
               
               
                   
                 11 
                 Slide19.jpg 
                 2123 
                 16.33 
               
               
                   
                 12 
                 Slide19.jpg 
                 1042 
                 8.02 
               
               
                   
                 13 
                 Slide19.jpg 
                 2105 
                   
               
               
                   
                 14 
                 Slide19.jpg 
                 957 
                 7.73 
               
               
                   
                 15 
                 Slide19.jpg 
                 308 
                 2.49 
               
               
                   
                 16 
                 Slide19.jpg 
                 1524 
                   
               
               
                   
                 17 
                 Slide19.jpg 
                 10 
                 0.05 
               
               
                   
                 18 
                 Slide19.jpg 
                 1530 
                 8.03 
               
               
                   
                 19 
                 Slide19.jpg 
                 2860 
                   
               
               
                   
                 20 
                 Slide19.jpg 
                 13 
                 0.10 
               
               
                   
                 21 
                 Slide19.jpg 
                 2293 
                 16.84 
               
               
                   
                 22 
                 Slide19.jpg 
                 3358 
                   
               
               
                   
                 23 
                 Slide19.jpg 
                 960 
                 10.58 
               
               
                   
                 24 
                 Slide19.jpg 
                 2639 
                 29.08 
               
               
                   
                 25 
                 Slide19.jpg 
                 2538 
                   
               
               
                   
                 26 
                 Slide19.jpg 
                 296 
                 3.03 
               
               
                   
                 27 
                 Slide19.jpg 
                 1797 
                 18.41 
               
               
                   
                 28 
                 Slide19.jpg 
                 1992 
                   
               
               
                   
                 29 
                 Slide19.jpg 
                 1105 
                 9.43 
               
               
                   
                 30 
                 Slide19.jpg 
                 401 
                 3.42 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 20 
               
               
                   
               
               
                 TNS3 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 36.39 
               
               
                   
                 IS 
                 61.20 
               
               
                   
                 LV 
                 97.58 
               
               
                   
                 IS/LV 
                 63% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 21 
               
               
                   
               
               
                 TNS4 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide20.jpg 
                 1524 
                   
               
               
                   
                  2 
                 Slide20.jpg 
                 47 
                 0.28 
               
               
                   
                  3 
                 Slide20.jpg 
                 1417 
                 8.37 
               
               
                   
                  4 
                 Slide20.jpg 
                 2478 
                   
               
               
                   
                  5 
                 Slide20.jpg 
                 582 
                 5.17 
               
               
                   
                  6 
                 Slide20.jpg 
                 1617 
                 14.36 
               
               
                   
                  7 
                 Slide20.jpg 
                 3284 
                   
               
               
                   
                  8 
                 Slide20.jpg 
                 1226 
                 11.20 
               
               
                   
                  9 
                 Slide20.jpg 
                 2072 
                 18.93 
               
               
                   
                 10 
                 Slide20.jpg 
                 3639 
                   
               
               
                   
                 11 
                 Slide20.jpg 
                 771 
                 7.20 
               
               
                   
                 12 
                 Slide20.jpg 
                 2177 
                 20.34 
               
               
                   
                 13 
                 Slide20.jpg 
                 3114 
                   
               
               
                   
                 14 
                 Slide20.jpg 
                 491 
                 5.36 
               
               
                   
                 15 
                 Slide20.jpg 
                 2189 
                 23.90 
               
               
                   
                 16 
                 Slide20.jpg 
                 1648 
                   
               
               
                   
                 17 
                 Slide20.jpg 
                 1244 
                 6.79 
               
               
                   
                 18 
                 Slide20.jpg 
                 94 
                 0.51 
               
               
                   
                 19 
                 Slide20.jpg 
                 2912 
                   
               
               
                   
                 20 
                 Slide20.jpg 
                 1446 
                 10.92 
               
               
                   
                 21 
                 Slide20.jpg 
                 1262 
                 9.53 
               
               
                   
                 22 
                 Slide20.jpg 
                 4073 
                   
               
               
                   
                 23 
                 Slide20.jpg 
                 2350 
                 17.31 
               
               
                   
                 24 
                 Slide20.jpg 
                 1049 
                 7.73 
               
               
                   
                 25 
                 Slide20.jpg 
                 3470 
                   
               
               
                   
                 26 
                 Slide20.jpg 
                 2445 
                 23.96 
               
               
                   
                 27 
                 Slide20.jpg 
                 1052 
                 10.31 
               
               
                   
                 28 
                 Slide20.jpg 
                 3219 
                   
               
               
                   
                 29 
                 Slide20.jpg 
                 2120 
                 22.39 
               
               
                   
                 30 
                 Slide20.jpg 
                 32 
                 0.34 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 22 
               
               
                   
               
               
                 TNS4 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 55.29 
               
               
                   
                 IS 
                 57.16 
               
               
                   
                 LV 
                 112.45 
               
               
                   
                 IS/LV 
                 51% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 23 
               
               
                   
               
               
                 TNF3 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide21.jpg 
                 1551 
                   
               
               
                   
                  2 
                 Slide21.jpg 
                 3 
                 0.02 
               
               
                   
                  3 
                 Slide21.jpg 
                 1502 
                 10.65 
               
               
                   
                  4 
                 Slide21.jpg 
                 3054 
                   
               
               
                   
                  5 
                 Slide21.jpg 
                 922 
                 6.34 
               
               
                   
                  6 
                 Slide21.jpg 
                 2049 
                 14.09 
               
               
                   
                  7 
                 Slide21.jpg 
                 3374 
                   
               
               
                   
                  8 
                 Slide21.jpg 
                 1280 
                 12.52 
               
               
                   
                  9 
                 Slide21.jpg 
                 1566 
                 15.32 
               
               
                   
                 10 
                 Slide21.jpg 
                 2799 
                   
               
               
                   
                 11 
                 Slide21.jpg 
                 1476 
                 14.77 
               
               
                   
                 12 
                 Slide21.jpg 
                 1061 
                 10.61 
               
               
                   
                 13 
                 Slide21.jpg 
                 2330 
                   
               
               
                   
                 14 
                 Slide21.jpg 
                 398 
                 3.25 
               
               
                   
                 15 
                 Slide21.jpg 
                 1012 
                 8.25 
               
               
                   
                 16 
                 Slide21.jpg 
                 1689 
                   
               
               
                   
                 17 
                 Slide21.jpg 
                 7 
                 0.05 
               
               
                   
                 18 
                 Slide21.jpg 
                 1544 
                 10.06 
               
               
                   
                 19 
                 Slide21.jpg 
                 2894 
                   
               
               
                   
                 20 
                 Slide21.jpg 
                 361 
                 2.62 
               
               
                   
                 21 
                 Slide21.jpg 
                 1925 
                 13.97 
               
               
                   
                 22 
                 Slide21.jpg 
                 3254 
                   
               
               
                   
                 23 
                 Slide21.jpg 
                 1137 
                 11.53 
               
               
                   
                 24 
                 Slide21.jpg 
                 1267 
                 12.85 
               
               
                   
                 25 
                 Slide21.jpg 
                 2814 
                   
               
               
                   
                 26 
                 Slide21.jpg 
                 1272 
                 12.66 
               
               
                   
                 27 
                 Slide21.jpg 
                 1113 
                 11.07 
               
               
                   
                 28 
                 Slide21.jpg 
                 2821 
                   
               
               
                   
                 29 
                 Slide21.jpg 
                 1438 
                 9.69 
               
               
                   
                 30 
                 Slide21.jpg 
                 174 
                 1.17 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 24 
               
               
                   
               
               
                 TNF3 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 36.71 
               
               
                   
                 IS 
                 54.02 
               
               
                   
                 LV 
                 90.74 
               
               
                   
                 IS/LV 
                 60% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 25 
               
               
                   
               
               
                 TNF4 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide22.jpg 
                 1354 
                   
               
               
                   
                  2 
                 Slide22.jpg 
                 72 
                 0.37 
               
               
                   
                  3 
                 Slide22.jpg 
                 1335 
                 6.90 
               
               
                   
                  4 
                 Slide22.jpg 
                 2892 
                   
               
               
                   
                  5 
                 Slide22.jpg 
                 672 
                 3.95 
               
               
                   
                  6 
                 Slide22.jpg 
                 2093 
                 12.30 
               
               
                   
                  7 
                 Slide22.jpg 
                 3414 
                   
               
               
                   
                  8 
                 Slide22.jpg 
                 1342 
                 9.83 
               
               
                   
                  9 
                 Slide22.jpg 
                 2213 
                 16.21 
               
               
                   
                 10 
                 Slide22.jpg 
                 3698 
                   
               
               
                   
                 11 
                 Slide22.jpg 
                 1168 
                 10.11 
               
               
                   
                 12 
                 Slide22.jpg 
                 2317 
                 20.05 
               
               
                   
                 13 
                 Slide22.jpg 
                 2565 
                   
               
               
                   
                 14 
                 Slide22.jpg 
                 243 
                 2.94 
               
               
                   
                 15 
                 Slide22.jpg 
                 1398 
                 16.90 
               
               
                   
                 16 
                 Slide22.jpg 
                 1486 
                   
               
               
                   
                 17 
                 Slide22.jpg 
                 638 
                 3.01 
               
               
                   
                 18 
                 Slide22.jpg 
                 583 
                 2.75 
               
               
                   
                 19 
                 Slide22.jpg 
                 2719 
                   
               
               
                   
                 20 
                 Slide22.jpg 
                 26 
                 0.16 
               
               
                   
                 21 
                 Slide22.jpg 
                 2164 
                 13.53 
               
               
                   
                 22 
                 Slide22.jpg 
                 3514 
                   
               
               
                   
                 23 
                 Slide22.jpg 
                 568 
                 4.04 
               
               
                   
                 24 
                 Slide22.jpg 
                 2361 
                 16.80 
               
               
                   
                 25 
                 Slide22.jpg 
                 3908 
                   
               
               
                   
                 26 
                 Slide22.jpg 
                 1498 
                 12.27 
               
               
                   
                 27 
                 Slide22.jpg 
                 1805 
                 14.78 
               
               
                   
                 28 
                 Slide22.jpg 
                 2946 
                   
               
               
                   
                 29 
                 Slide22.jpg 
                 16 
                 0.17 
               
               
                   
                 30 
                 Slide22.jpg 
                 1969 
                 20.72 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 26 
               
               
                   
               
               
                 TNF4 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 23.42 
               
               
                   
                 IS 
                 70.46 
               
               
                   
                 LV 
                 93.88 
               
               
                   
                 IS/LV 
                 75% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 27 
               
               
                   
               
               
                 TNS5 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide23.jpg 
                 1615 
                   
               
               
                   
                  2 
                 Slide23.jpg 
                 8 
                 0.04 
               
               
                   
                  3 
                 Slide23.jpg 
                 1571 
                 8.75 
               
               
                   
                  4 
                 Slide23.jpg 
                 2789 
                   
               
               
                   
                  5 
                 Slide23.jpg 
                 1477 
                 11.65 
               
               
                   
                  6 
                 Slide23.jpg 
                 1042 
                 8.22 
               
               
                   
                  7 
                 Slide23.jpg 
                 3558 
                   
               
               
                   
                  8 
                 Slide23.jpg 
                 2026 
                 22.21 
               
               
                   
                  9 
                 Slide23.jpg 
                 1327 
                 14.55 
               
               
                   
                 10 
                 Slide23.jpg 
                 3822 
                   
               
               
                   
                 11 
                 Slide23.jpg 
                 1044 
                 8.74 
               
               
                   
                 12 
                 Slide23.jpg 
                 1590 
                 13.31 
               
               
                   
                 13 
                 Slide23.jpg 
                 3246 
                   
               
               
                   
                 14 
                 Slide23.jpg 
                 1224 
                 8.67 
               
               
                   
                 15 
                 Slide23.jpg 
                 705 
                 5.00 
               
               
                   
                 16 
                 Slide23.jpg 
                 1445 
                   
               
               
                   
                 17 
                 Slide23.jpg 
                 1228 
                 7.65 
               
               
                   
                 18 
                 Slide23.jpg 
                 200 
                 1.25 
               
               
                   
                 19 
                 Slide23.jpg 
                 2732 
                   
               
               
                   
                 20 
                 Slide23.jpg 
                 1951 
                 15.71 
               
               
                   
                 21 
                 Slide23.jpg 
                 782 
                 6.30 
               
               
                   
                 22 
                 Slide23.jpg 
                 3858 
                   
               
               
                   
                 23 
                 Slide23.jpg 
                 3039 
                 30.72 
               
               
                   
                 24 
                 Slide23.jpg 
                 400 
                 4.04 
               
               
                   
                 25 
                 Slide23.jpg 
                 3697 
                   
               
               
                   
                 26 
                 Slide23.jpg 
                 2609 
                 22.58 
               
               
                   
                 27 
                 Slide23.jpg 
                 943 
                 8.16 
               
               
                   
                 28 
                 Slide23.jpg 
                 3358 
                   
               
               
                   
                 29 
                 Slide23.jpg 
                 1492 
                 10.22 
               
               
                   
                 30 
                 Slide23.jpg 
                 583 
                 3.99 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 28 
               
               
                   
               
               
                 TNS5 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 69.10 
               
               
                   
                 IS 
                 36.78 
               
               
                   
                 LV 
                 105.88 
               
               
                   
                 IS/LV 
                 35% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 29 
               
               
                   
               
               
                 TNS6 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide24.jpg 
                 1216 
                   
               
               
                   
                  2 
                 Slide24.jpg 
                 258 
                 1.49 
               
               
                   
                  3 
                 Slide24.jpg 
                 770 
                 4.43 
               
               
                   
                  4 
                 Slide24.jpg 
                 3079 
                   
               
               
                   
                  5 
                 Slide24.jpg 
                 1436 
                 10.26 
               
               
                   
                  6 
                 Slide24.jpg 
                 1417 
                 10.12 
               
               
                   
                  7 
                 Slide24.jpg 
                 3677 
                   
               
               
                   
                  8 
                 Slide24.jpg 
                 2085 
                 11.34 
               
               
                   
                  9 
                 Slide24.jpg 
                 1122 
                 6.10 
               
               
                   
                 10 
                 Slide24.jpg 
                 3908 
                   
               
               
                   
                 11 
                 Slide24.jpg 
                 2151 
                 15.96 
               
               
                   
                 12 
                 Slide24.jpg 
                 1415 
                 10.50 
               
               
                   
                 13 
                 Slide24.jpg 
                 2371 
                   
               
               
                   
                 14 
                 Slide24.jpg 
                 1651 
                 14.62 
               
               
                   
                 15 
                 Slide24.jpg 
                 495 
                 4.38 
               
               
                   
                 16 
                 Slide24.jpg 
                 1123 
                   
               
               
                   
                 17 
                 Slide24.jpg 
                 879 
                 5.48 
               
               
                   
                 18 
                 Slide24.jpg 
                 262 
                 1.63 
               
               
                   
                 19 
                 Slide24.jpg 
                 3090 
                   
               
               
                   
                 20 
                 Slide24.jpg 
                 1775 
                 12.64 
               
               
                   
                 21 
                 Slide24.jpg 
                 1121 
                 7.98 
               
               
                   
                 22 
                 Slide24.jpg 
                 3470 
                   
               
               
                   
                 23 
                 Slide24.jpg 
                 2215 
                 12.77 
               
               
                   
                 24 
                 Slide24.jpg 
                 1219 
                 7.03 
               
               
                   
                 25 
                 Slide24.jpg 
                 3666 
                   
               
               
                   
                 26 
                 Slide24.jpg 
                 2524 
                 19.97 
               
               
                   
                 27 
                 Slide24.jpg 
                 1411 
                 11.16 
               
               
                   
                 28 
                 Slide24.jpg 
                 2470 
                   
               
               
                   
                 29 
                 Slide24.jpg 
                 1397 
                 11.88 
               
               
                   
                 30 
                 Slide24.jpg 
                 140 
                 1.19 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 30 
               
               
                   
               
               
                 TNS6 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 58.20 
               
               
                   
                 IS 
                 32.27 
               
               
                   
                 LV 
                 90.47 
               
               
                   
                 IS/LV 
                 36% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 31 
               
               
                   
               
               
                 CN13 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide25.jpg 
                 1010 
                   
               
               
                   
                  2 
                 Slide25.jpg 
                 4 
                 0.04 
               
               
                   
                  3 
                 Slide25.jpg 
                 1006 
                 8.96 
               
               
                   
                  4 
                 Slide25.jpg 
                 2216 
                   
               
               
                   
                  5 
                 Slide25.jpg 
                 756 
                 5.80 
               
               
                   
                  6 
                 Slide25.jpg 
                 1708 
                 13.10 
               
               
                   
                  7 
                 Slide25.jpg 
                 3122 
                   
               
               
                   
                  8 
                 Slide25.jpg 
                 744 
                 5.72 
               
               
                   
                  9 
                 Slide25.jpg 
                 1674 
                 12.87 
               
               
                   
                 10 
                 Slide25.jpg 
                 3214 
                   
               
               
                   
                 11 
                 Slide25.jpg 
                 177 
                 1.87 
               
               
                   
                 12 
                 Slide25.jpg 
                 1678 
                 17.75 
               
               
                   
                 13 
                 Slide25.jpg 
                 2504 
                   
               
               
                   
                 14 
                 Slide25.jpg 
                 371 
                 3.41 
               
               
                   
                 15 
                 Slide25.jpg 
                 770 
                 7.07 
               
               
                   
                 16 
                 Slide25.jpg 
                 940 
                   
               
               
                   
                 17 
                 Slide25.jpg 
                 3 
                 0.03 
               
               
                   
                 18 
                 Slide25.jpg 
                 902 
                 8.64 
               
               
                   
                 19 
                 Slide25.jpg 
                 1907 
                   
               
               
                   
                 20 
                 Slide25.jpg 
                 266 
                 2.37 
               
               
                   
                 21 
                 Slide25.jpg 
                 1439 
                 12.83 
               
               
                   
                 22 
                 Slide25.jpg 
                 2763 
                   
               
               
                   
                 23 
                 Slide25.jpg 
                 1036 
                 9.00 
               
               
                   
                 24 
                 Slide25.jpg 
                 1855 
                 16.11 
               
               
                   
                 25 
                 Slide25.jpg 
                 2930 
                   
               
               
                   
                 26 
                 Slide25.jpg 
                 988 
                 11.46 
               
               
                   
                 27 
                 Slide25.jpg 
                 1618 
                 18.78 
               
               
                   
                 28 
                 Slide25.jpg 
                 2498 
                   
               
               
                   
                 29 
                 Slide25.jpg 
                 280 
                 2.58 
               
               
                   
                 30 
                 Slide25.jpg 
                 1839 
                 16.93 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 32 
               
               
                   
               
               
                 CN13 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 21.14 
               
               
                   
                 IS 
                 66.52 
               
               
                   
                 LV 
                 87.66 
               
               
                   
                 IS/LV 
                 76% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 33 
               
               
                   
               
               
                 CN14 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide26.jpg 
                 1387 
                   
               
               
                   
                  2 
                 Slide26.jpg 
                 40 
                 0.23 
               
               
                   
                  3 
                 Slide26.jpg 
                 1356 
                 7.82 
               
               
                   
                  4 
                 Slide26.jpg 
                 2994 
                   
               
               
                   
                  5 
                 Slide26.jpg 
                 699 
                 4.67 
               
               
                   
                  6 
                 Slide26.jpg 
                 1620 
                 10.82 
               
               
                   
                  7 
                 Slide26.jpg 
                 3017 
                   
               
               
                   
                  8 
                 Slide26.jpg 
                 1087 
                 11.89 
               
               
                   
                  9 
                 Slide26.jpg 
                 1443 
                 15.78 
               
               
                   
                 10 
                 Slide26.jpg 
                 2871 
                   
               
               
                   
                 11 
                 Slide26.jpg 
                 2644 
                 29.47 
               
               
                   
                 12 
                 Slide26.jpg 
                 188 
                 2.10 
               
               
                   
                 13 
                 Slide26.jpg 
                 2504 
                   
               
               
                   
                 14 
                 Slide26.jpg 
                 7 
                 0.05 
               
               
                   
                 15 
                 Slide26.jpg 
                 1996 
                 13.55 
               
               
                   
                 16 
                 Slide26.jpg 
                 1424 
                   
               
               
                   
                 17 
                 Slide26.jpg 
                 490 
                 2.75 
               
               
                   
                 18 
                 Slide26.jpg 
                 931 
                 5.23 
               
               
                   
                 19 
                 Slide26.jpg 
                 2926 
                   
               
               
                   
                 20 
                 Slide26.jpg 
                 40 
                 0.27 
               
               
                   
                 21 
                 Slide26.jpg 
                 2231 
                 15.25 
               
               
                   
                 22 
                 Slide26.jpg 
                 3248 
                   
               
               
                   
                 23 
                 Slide26.jpg 
                 782 
                 7.95 
               
               
                   
                 24 
                 Slide26.jpg 
                 2137 
                 21.71 
               
               
                   
                 25 
                 Slide26.jpg 
                 3401 
                   
               
               
                   
                 26 
                 Slide26.jpg 
                 348 
                 3.27 
               
               
                   
                 27 
                 Slide26.jpg 
                 2624 
                 24.69 
               
               
                   
                 28 
                 Slide26.jpg 
                 2079 
                   
               
               
                   
                 29 
                 Slide26.jpg 
                 573 
                 4.69 
               
               
                   
                 30 
                 Slide26.jpg 
                 1042 
                 8.52 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 34 
               
               
                   
               
               
                 CN14 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 32.62 
               
               
                   
                 IS 
                 62.74 
               
               
                   
                 LV 
                 95.36 
               
               
                   
                 IS/LV 
                 66% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 35 
               
               
                   
               
               
                 TNF5 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide27.jpg 
                 1504 
                   
               
               
                   
                  2 
                 Slide27.jpg 
                 22 
                 0.13 
               
               
                   
                  3 
                 Slide27.jpg 
                 1336 
                 7.99 
               
               
                   
                  4 
                 Slide27.jpg 
                 2786 
                   
               
               
                   
                  5 
                 Slide27.jpg 
                 390 
                 3.22 
               
               
                   
                  6 
                 Slide27.jpg 
                 1956 
                 16.15 
               
               
                   
                  7 
                 Slide27.jpg 
                 3792 
                   
               
               
                   
                  8 
                 Slide27.jpg 
                 1444 
                 10.66 
               
               
                   
                  9 
                 Slide27.jpg 
                 2232 
                 16.48 
               
               
                   
                 10 
                 Slide27.jpg 
                 3470 
                   
               
               
                   
                 11 
                 Slide27.jpg 
                 587 
                 5.41 
               
               
                   
                 12 
                 Slide27.jpg 
                 2824 
                 26.04 
               
               
                   
                 13 
                 Slide27.jpg 
                 3002 
                   
               
               
                   
                 14 
                 Slide27.jpg 
                 2361 
                 16.52 
               
               
                   
                 15 
                 Slide27.jpg 
                 1329 
                 9.30 
               
               
                   
                 16 
                 Slide27.jpg 
                 1666 
                   
               
               
                   
                 17 
                 Slide27.jpg 
                 274 
                 1.48 
               
               
                   
                 18 
                 Slide27.jpg 
                 1024 
                 5.53 
               
               
                   
                 19 
                 Slide27.jpg 
                 2735 
                   
               
               
                   
                 20 
                 Slide27.jpg 
                 9 
                 0.08 
               
               
                   
                 21 
                 Slide27.jpg 
                 2897 
                 24.36 
               
               
                   
                 22 
                 Slide27.jpg 
                 3575 
                   
               
               
                   
                 23 
                 Slide27.jpg 
                 1217 
                 9.53 
               
               
                   
                 24 
                 Slide27.jpg 
                 2163 
                 16.94 
               
               
                   
                 25 
                 Slide27.jpg 
                 3350 
                   
               
               
                   
                 26 
                 Slide27.jpg 
                 997 
                 9.52 
               
               
                   
                 27 
                 Slide27.jpg 
                 1812 
                 17.31 
               
               
                   
                 28 
                 Slide27.jpg 
                 3022 
                   
               
               
                   
                 29 
                 Slide27.jpg 
                 12 
                 0.08 
               
               
                   
                 30 
                 Slide27.jpg 
                 1778 
                 12.36 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 36 
               
               
                   
               
               
                 TNF5 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 28.32 
               
               
                   
                 IS 
                 76.23 
               
               
                   
                 LV 
                 104.55 
               
               
                   
                 IS/LV 
                 73% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 37 
               
               
                   
               
               
                 TNF6 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide28.jpg 
                 1114 
                   
               
               
                   
                  2 
                 Slide28.jpg 
                 62 
                 0.45 
               
               
                   
                  3 
                 Slide28.jpg 
                 879 
                 6.31 
               
               
                   
                  4 
                 Slide28.jpg 
                 2858 
                   
               
               
                   
                  5 
                 Slide28.jpg 
                 459 
                 3.85 
               
               
                   
                  6 
                 Slide28.jpg 
                 1713 
                 14.38 
               
               
                   
                  7 
                 Slide28.jpg 
                 3625 
                   
               
               
                   
                  8 
                 Slide28.jpg 
                 369 
                 3.56 
               
               
                   
                  9 
                 Slide28.jpg 
                 2924 
                 28.23 
               
               
                   
                 10 
                 Slide28.jpg 
                 3948 
                   
               
               
                   
                 11 
                 Slide28.jpg 
                 511 
                 4.27 
               
               
                   
                 12 
                 Slide28.jpg 
                 2866 
                 23.96 
               
               
                   
                 13 
                 Slide28.jpg 
                 3135 
                   
               
               
                   
                 14 
                 Slide28.jpg 
                 386 
                 3.08 
               
               
                   
                 15 
                 Slide28.jpg 
                 1447 
                 11.54 
               
               
                   
                 16 
                 Slide28.jpg 
                 1126 
                   
               
               
                   
                 17 
                 Slide28.jpg 
                 10 
                 0.07 
               
               
                   
                 18 
                 Slide28.jpg 
                 1043 
                 7.41 
               
               
                   
                 19 
                 Slide28.jpg 
                 3156 
                   
               
               
                   
                 20 
                 Slide28.jpg 
                 160 
                 1.22 
               
               
                   
                 21 
                 Slide28.jpg 
                 3062 
                 23.29 
               
               
                   
                 22 
                 Slide28.jpg 
                 3790 
                   
               
               
                   
                 23 
                 Slide28.jpg 
                 827 
                 7.64 
               
               
                   
                 24 
                 Slide28.jpg 
                 2644 
                 24.42 
               
               
                   
                 25 
                 Slide28.jpg 
                 3618 
                   
               
               
                   
                 26 
                 Slide28.jpg 
                 1607 
                 14.66 
               
               
                   
                 27 
                 Slide28.jpg 
                 2452 
                 22.36 
               
               
                   
                 28 
                 Slide28.jpg 
                 3440 
                   
               
               
                   
                 29 
                 Slide28.jpg 
                 1023 
                 7.43 
               
               
                   
                 30 
                 Slide28.jpg 
                 1770 
                 12.86 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 38 
               
               
                   
               
               
                 TNF6 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 23.11 
               
               
                   
                 IS 
                 87.38 
               
               
                   
                 LV 
                 110.50 
               
               
                   
                 IS/LV 
                 79% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 39 
               
               
                   
               
               
                 TNS7 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide29.jpg 
                 1713 
                   
               
               
                   
                  2 
                 Slide29.jpg 
                 607 
                 4.61 
               
               
                   
                  3 
                 Slide29.jpg 
                 782 
                 5.93 
               
               
                   
                  4 
                 Slide29.jpg 
                 2484 
                   
               
               
                   
                  5 
                 Slide29.jpg 
                 195 
                 1.88 
               
               
                   
                  6 
                 Slide29.jpg 
                 1842 
                 17.80 
               
               
                   
                  7 
                 Slide29.jpg 
                 2807 
                   
               
               
                   
                  8 
                 Slide29.jpg 
                 1568 
                 12.29 
               
               
                   
                  9 
                 Slide29.jpg 
                 380 
                 2.98 
               
               
                   
                 10 
                 Slide29.jpg 
                 3271 
                   
               
               
                   
                 11 
                 Slide29.jpg 
                 2187 
                 20.06 
               
               
                   
                 12 
                 Slide29.jpg 
                 350 
                 3.21 
               
               
                   
                 13 
                 Slide29.jpg 
                 2309 
                   
               
               
                   
                 14 
                 Slide29.jpg 
                 610 
                 5.55 
               
               
                   
                 15 
                 Slide29.jpg 
                 1008 
                 9.17 
               
               
                   
                 16 
                 Slide29.jpg 
                 1923 
                   
               
               
                   
                 17 
                 Slide29.jpg 
                 865 
                 5.85 
               
               
                   
                 18 
                 Slide29.jpg 
                 631 
                 4.27 
               
               
                   
                 19 
                 Slide29.jpg 
                 3033 
                   
               
               
                   
                 20 
                 Slide29.jpg 
                 1501 
                 11.88 
               
               
                   
                 21 
                 Slide29.jpg 
                 780 
                 6.17 
               
               
                   
                 22 
                 Slide29.jpg 
                 3287 
                   
               
               
                   
                 23 
                 Slide29.jpg 
                 2214 
                 14.82 
               
               
                   
                 24 
                 Slide29.jpg 
                 456 
                 3.05 
               
               
                   
                 25 
                 Slide29.jpg 
                 3395 
                   
               
               
                   
                 26 
                 Slide29.jpg 
                 2398 
                 21.19 
               
               
                   
                 27 
                 Slide29.jpg 
                 287 
                 2.54 
               
               
                   
                 28 
                 Slide29.jpg 
                 2969 
                   
               
               
                   
                 29 
                 Slide29.jpg 
                 1647 
                 11.65 
               
               
                   
                 30 
                 Slide29.jpg 
                 67 
                 0.47 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 40 
               
               
                   
               
               
                 TNS7 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 54.88 
               
               
                   
                 IS 
                 27.79 
               
               
                   
                 LV 
                 82.68 
               
               
                   
                 IS/LV 
                 34% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 41 
               
               
                   
               
               
                 TNS8 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide30.jpg 
                 1123 
                   
               
               
                   
                  2 
                 Slide30.jpg 
                 11 
                 0.05 
               
               
                   
                  3 
                 Slide30.jpg 
                 988 
                 4.40 
               
               
                   
                  4 
                 Slide30.jpg 
                 2352 
                   
               
               
                   
                  5 
                 Slide30.jpg 
                 279 
                 2.25 
               
               
                   
                  6 
                 Slide30.jpg 
                 2001 
                 16.16 
               
               
                   
                  7 
                 Slide30.jpg 
                 3274 
                   
               
               
                   
                  8 
                 Slide30.jpg 
                 1085 
                 7.29 
               
               
                   
                  9 
                 Slide30.jpg 
                 1821 
                 12.24 
               
               
                   
                 10 
                 Slide30.jpg 
                 3333 
                   
               
               
                   
                 11 
                 Slide30.jpg 
                 2048 
                 17.20 
               
               
                   
                 12 
                 Slide30.jpg 
                 838 
                 7.04 
               
               
                   
                 13 
                 Slide30.jpg 
                 2240 
                   
               
               
                   
                 14 
                 Slide30.jpg 
                 793 
                 7.08 
               
               
                   
                 15 
                 Slide30.jpg 
                 840 
                 7.50 
               
               
                   
                 16 
                 Slide30.jpg 
                 914 
                   
               
               
                   
                 17 
                 Slide30.jpg 
                 866 
                 4.74 
               
               
                   
                 18 
                 Slide30.jpg 
                 64 
                 0.35 
               
               
                   
                 19 
                 Slide30.jpg 
                 2811 
                   
               
               
                   
                 20 
                 Slide30.jpg 
                 397 
                 2.68 
               
               
                   
                 21 
                 Slide30.jpg 
                 2135 
                 14.43 
               
               
                   
                 22 
                 Slide30.jpg 
                 3378 
                   
               
               
                   
                 23 
                 Slide30.jpg 
                 588 
                 3.83 
               
               
                   
                 24 
                 Slide30.jpg 
                 2250 
                 14.65 
               
               
                   
                 25 
                 Slide30.jpg 
                 3241 
                   
               
               
                   
                 26 
                 Slide30.jpg 
                 2671 
                 23.08 
               
               
                   
                 27 
                 Slide30.jpg 
                 287 
                 2.48 
               
               
                   
                 28 
                 Slide30.jpg 
                 2697 
                   
               
               
                   
                 29 
                 Slide30.jpg 
                 1247 
                 9.25 
               
               
                   
                 30 
                 Slide30.jpg 
                 23 
                 0.17 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 42 
               
               
                   
               
               
                 TNS8 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 38.73 
               
               
                   
                 IS 
                 39.71 
               
               
                   
                 LV 
                 78.44 
               
               
                   
                 IS/LV 
                 51% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 43 
               
               
                   
               
               
                 TNF7 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide31.jpg 
                 1733 
                   
               
               
                   
                  2 
                 Slide31.jpg 
                 15 
                 0.06 
               
               
                   
                  3 
                 Slide31.jpg 
                 1704 
                 6.88 
               
               
                   
                  4 
                 Slide31.jpg 
                 3401 
                   
               
               
                   
                  5 
                 Slide31.jpg 
                 719 
                 3.38 
               
               
                   
                  6 
                 Slide31.jpg 
                 2216 
                 10.43 
               
               
                   
                  7 
                 Slide31.jpg 
                 3789 
                   
               
               
                   
                  8 
                 Slide31.jpg 
                 917 
                 7.02 
               
               
                   
                  9 
                 Slide31.jpg 
                 2163 
                 16.56 
               
               
                   
                 10 
                 Slide31.jpg 
                 4149 
                   
               
               
                   
                 11 
                 Slide31.jpg 
                 719 
                 5.03 
               
               
                   
                 12 
                 Slide31.jpg 
                 3423 
                 23.93 
               
               
                   
                 13 
                 Slide31.jpg 
                 3309 
                   
               
               
                   
                 14 
                 Slide31.jpg 
                 1479 
                 8.49 
               
               
                   
                 15 
                 Slide31.jpg 
                 1771 
                 10.17 
               
               
                   
                 16 
                 Slide31.jpg 
                 1777 
                   
               
               
                   
                 17 
                 Slide31.jpg 
                 1049 
                 4.13 
               
               
                   
                 18 
                 Slide31.jpg 
                 678 
                 2.67 
               
               
                   
                 19 
                 Slide31.jpg 
                 3117 
                   
               
               
                   
                 20 
                 Slide31.jpg 
                 221 
                 1.13 
               
               
                   
                 21 
                 Slide31.jpg 
                 2281 
                 11.71 
               
               
                   
                 22 
                 Slide31.jpg 
                 3970 
                   
               
               
                   
                 23 
                 Slide31.jpg 
                 2416 
                 17.65 
               
               
                   
                 24 
                 Slide31.jpg 
                 796 
                 5.81 
               
               
                   
                 25 
                 Slide31.jpg 
                 4354 
                   
               
               
                   
                 26 
                 Slide31.jpg 
                 3291 
                 21.92 
               
               
                   
                 27 
                 Slide31.jpg 
                 697 
                 4.64 
               
               
                   
                 28 
                 Slide31.jpg 
                 3316 
                   
               
               
                   
                 29 
                 Slide31.jpg 
                 2414 
                 13.83 
               
               
                   
                 30 
                 Slide31.jpg 
                 62 
                 0.36 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 44 
               
               
                   
               
               
                 TNF7 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 41.32 
               
               
                   
                 IS 
                 46.57 
               
               
                   
                 LV 
                 87.90 
               
               
                   
                 IS/LV 
                 53% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 45 
               
               
                   
               
               
                 TNF8 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide32.jpg 
                 1553 
                   
               
               
                   
                  2 
                 Slide32.jpg 
                 572 
                 2.58 
               
               
                   
                  3 
                 Slide32.jpg 
                 873 
                 3.93 
               
               
                   
                  4 
                 Slide32.jpg 
                 3334 
                   
               
               
                   
                  5 
                 Slide32.jpg 
                 1084 
                 5.53 
               
               
                   
                  6 
                 Slide32.jpg 
                 1525 
                 7.78 
               
               
                   
                  7 
                 Slide32.jpg 
                 4166 
                   
               
               
                   
                  8 
                 Slide32.jpg 
                 2437 
                 12.87 
               
               
                   
                  9 
                 Slide32.jpg 
                 1557 
                 8.22 
               
               
                   
                 10 
                 Slide32.jpg 
                 4558 
                   
               
               
                   
                 11 
                 Slide32.jpg 
                 2698 
                 20.13 
               
               
                   
                 12 
                 Slide32.jpg 
                 1306 
                 9.74 
               
               
                   
                 13 
                 Slide32.jpg 
                 3405 
                   
               
               
                   
                 14 
                 Slide32.jpg 
                 2991 
                 25.47 
               
               
                   
                 15 
                 Slide32.jpg 
                 51 
                 0.43 
               
               
                   
                 16 
                 Slide32.jpg 
                 1543 
                   
               
               
                   
                 17 
                 Slide32.jpg 
                 3 
                 0.01 
               
               
                   
                 18 
                 Slide32.jpg 
                 1407 
                 6.38 
               
               
                   
                 19 
                 Slide32.jpg 
                 3359 
                   
               
               
                   
                 20 
                 Slide32.jpg 
                 581 
                 2.94 
               
               
                   
                 21 
                 Slide32.jpg 
                 2011 
                 10.18 
               
               
                   
                 22 
                 Slide32.jpg 
                 3986 
                   
               
               
                   
                 23 
                 Slide32.jpg 
                 202 
                 1.11 
               
               
                   
                 24 
                 Slide32.jpg 
                 3788 
                 20.91 
               
               
                   
                 25 
                 Slide32.jpg 
                 4684 
                   
               
               
                   
                 26 
                 Slide32.jpg 
                 425 
                 3.08 
               
               
                   
                 27 
                 Slide32.jpg 
                 3308 
                 24.01 
               
               
                   
                 28 
                 Slide32.jpg 
                 3498 
                   
               
               
                   
                 29 
                 Slide32.jpg 
                 920 
                 7.63 
               
               
                   
                 30 
                 Slide32.jpg 
                 1731 
                 14.35 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 46 
               
               
                   
               
               
                 TNF8 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 40.68 
               
               
                   
                 IS 
                 52.97 
               
               
                   
                 LV 
                 93.65 
               
               
                   
                 IS/LV 
                 57% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 47 
               
               
                   
               
               
                 TNS9 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide33.jpg 
                 2637 
                   
               
               
                   
                  2 
                 Slide33.jpg 
                 14 
                 0.06 
               
               
                   
                  3 
                 Slide33.jpg 
                 2081 
                 9.47 
               
               
                   
                  4 
                 Slide33.jpg 
                 4101 
                   
               
               
                   
                  5 
                 Slide33.jpg 
                 1571 
                 7.28 
               
               
                   
                  6 
                 Slide33.jpg 
                 1516 
                 7.02 
               
               
                   
                  7 
                 Slide33.jpg 
                 4527 
                   
               
               
                   
                  8 
                 Slide33.jpg 
                 2519 
                 18.36 
               
               
                   
                  9 
                 Slide33.jpg 
                 1555 
                 11.34 
               
               
                   
                 10 
                 Slide33.jpg 
                 3326 
                   
               
               
                   
                 11 
                 Slide33.jpg 
                 3188 
                 19.17 
               
               
                   
                 12 
                 Slide33.jpg 
                 27 
                 0.16 
               
               
                   
                 13 
                 Slide33.jpg 
                 2336 
                   
               
               
                   
                 14 
                 Slide33.jpg 
                 1885 
                 9.68 
               
               
                   
                 15 
                 Slide33.jpg 
                 240 
                 1.23 
               
               
                   
                 16 
                 Slide33.jpg 
                 2343 
                   
               
               
                   
                 17 
                 Slide33.jpg 
                 2027 
                 10.38 
               
               
                   
                 18 
                 Slide33.jpg 
                 21 
                 0.11 
               
               
                   
                 19 
                 Slide33.jpg 
                 3393 
                   
               
               
                   
                 20 
                 Slide33.jpg 
                 1928 
                 10.80 
               
               
                   
                 21 
                 Slide33.jpg 
                 945 
                 5.29 
               
               
                   
                 22 
                 Slide33.jpg 
                 4425 
                   
               
               
                   
                 23 
                 Slide33.jpg 
                 2984 
                 22.25 
               
               
                   
                 24 
                 Slide33.jpg 
                 637 
                 4.75 
               
               
                   
                 25 
                 Slide33.jpg 
                 3063 
                   
               
               
                   
                 26 
                 Slide33.jpg 
                 773 
                 5.05 
               
               
                   
                 27 
                 Slide33.jpg 
                 1885 
                 12.31 
               
               
                   
                 28 
                 Slide33.jpg 
                 2324 
                   
               
               
                   
                 29 
                 Slide33.jpg 
                 1390 
                 7.18 
               
               
                   
                 30 
                 Slide33.jpg 
                 9 
                 0.05 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 48 
               
               
                   
               
               
                 TNS9 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 55.11 
               
               
                   
                 IS 
                 25.86 
               
               
                   
                 LV 
                 80.97 
               
               
                   
                 IS/LV 
                 32% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 49 
               
               
                   
               
               
                 TNS10 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide34.jpg 
                 1775 
                   
               
               
                   
                  2 
                 Slide34.jpg 
                 1082 
                 4.88 
               
               
                   
                  3 
                 Slide34.jpg 
                 348 
                 1.57 
               
               
                   
                  4 
                 Slide34.jpg 
                 3607 
                   
               
               
                   
                  5 
                 Slide34.jpg 
                 1823 
                 11.12 
               
               
                   
                  6 
                 Slide34.jpg 
                 1483 
                 9.05 
               
               
                   
                  7 
                 Slide34.jpg 
                 4313 
                   
               
               
                   
                  8 
                 Slide34.jpg 
                 1087 
                 6.80 
               
               
                   
                  9 
                 Slide34.jpg 
                 2173 
                 13.60 
               
               
                   
                 10 
                 Slide34.jpg 
                 4275 
                   
               
               
                   
                 11 
                 Slide34.jpg 
                 2471 
                 15.03 
               
               
                   
                 12 
                 Slide34.jpg 
                 1734 
                 10.55 
               
               
                   
                 13 
                 Slide34.jpg 
                 2864 
                   
               
               
                   
                 14 
                 Slide34.jpg 
                 2424 
                 18.62 
               
               
                   
                 15 
                 Slide34.jpg 
                 43 
                 0.33 
               
               
                   
                 16 
                 Slide34.jpg 
                 1601 
                   
               
               
                   
                 17 
                 Slide34.jpg 
                 1600 
                 8.00 
               
               
                   
                 18 
                 Slide34.jpg 
                 16 
                 0.08 
               
               
                   
                 19 
                 Slide34.jpg 
                 3486 
                   
               
               
                   
                 20 
                 Slide34.jpg 
                 933 
                 5.89 
               
               
                   
                 21 
                 Slide34.jpg 
                 935 
                 5.90 
               
               
                   
                 22 
                 Slide34.jpg 
                 4312 
                   
               
               
                   
                 23 
                 Slide34.jpg 
                 3250 
                 20.35 
               
               
                   
                 24 
                 Slide34.jpg 
                 722 
                 4.52 
               
               
                   
                 25 
                 Slide34.jpg 
                 4178 
                   
               
               
                   
                 26 
                 Slide34.jpg 
                 3996 
                 24.87 
               
               
                   
                 27 
                 Slide34.jpg 
                 231 
                 1.44 
               
               
                   
                 28 
                 Slide34.jpg 
                 3046 
                   
               
               
                   
                 29 
                 Slide34.jpg 
                 2854 
                 20.61 
               
               
                   
                 30 
                 Slide34.jpg 
                 39 
                 0.28 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 50 
               
               
                   
               
               
                 TNS10 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 68.08 
               
               
                   
                 IS 
                 23.66 
               
               
                   
                 LV 
                 91.74 
               
               
                   
                 IS/LV 
                 26% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 51 
               
               
                   
               
               
                 TNF9 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide35.jpg 
                 1737 
                   
               
               
                   
                  2 
                 Slide35.jpg 
                 841 
                 2.91 
               
               
                   
                  3 
                 Slide35.jpg 
                 788 
                 2.72 
               
               
                   
                  4 
                 Slide35.jpg 
                 3368 
                   
               
               
                   
                  5 
                 Slide35.jpg 
                 1416 
                 7.99 
               
               
                   
                  6 
                 Slide35.jpg 
                 1230 
                 6.94 
               
               
                   
                  7 
                 Slide35.jpg 
                 4474 
                   
               
               
                   
                  8 
                 Slide35.jpg 
                 1046 
                 8.18 
               
               
                   
                  9 
                 Slide35.jpg 
                 3356 
                 26.25 
               
               
                   
                 10 
                 Slide35.jpg 
                 4877 
                   
               
               
                   
                 11 
                 Slide35.jpg 
                 1303 
                 6.68 
               
               
                   
                 12 
                 Slide35.jpg 
                 3142 
                 16.11 
               
               
                   
                 13 
                 Slide35.jpg 
                 3803 
                   
               
               
                   
                 14 
                 Slide35.jpg 
                 2906 
                 16.81 
               
               
                   
                 15 
                 Slide35.jpg 
                 15 
                 0.09 
               
               
                   
                 16 
                 Slide35.jpg 
                 1719 
                   
               
               
                   
                 17 
                 Slide35.jpg 
                 8 
                 0.03 
               
               
                   
                 18 
                 Slide35.jpg 
                 1545 
                 5.39 
               
               
                   
                 19 
                 Slide35.jpg 
                 3500 
                   
               
               
                   
                 20 
                 Slide35.jpg 
                 9 
                 0.05 
               
               
                   
                 21 
                 Slide35.jpg 
                 3382 
                 18.36 
               
               
                   
                 22 
                 Slide35.jpg 
                 4790 
                   
               
               
                   
                 23 
                 Slide35.jpg 
                 9 
                 0.07 
               
               
                   
                 24 
                 Slide35.jpg 
                 4476 
                 32.71 
               
               
                   
                 25 
                 Slide35.jpg 
                 4213 
                   
               
               
                   
                 26 
                 Slide35.jpg 
                 1798 
                 10.67 
               
               
                   
                 27 
                 Slide35.jpg 
                 2840 
                 16.85 
               
               
                   
                 28 
                 Slide35.jpg 
                 3714 
                   
               
               
                   
                 29 
                 Slide35.jpg 
                 2917 
                 17.28 
               
               
                   
                 30 
                 Slide35.jpg 
                 342 
                 2.03 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 52 
               
               
                   
               
               
                 TNF9 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 35.33 
               
               
                   
                 IS 
                 63.72 
               
               
                   
                 LV 
                 99.05 
               
               
                   
                 IS/LV 
                 64% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 53 
               
               
                   
               
               
                 TNF10 raw values 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                  1 
                 Slide36.jpg 
                 2294 
                   
               
               
                   
                  2 
                 Slide36.jpg 
                 14 
                 0.08 
               
               
                   
                  3 
                 Slide36.jpg 
                 2183 
                 12.37 
               
               
                   
                  4 
                 Slide36.jpg 
                 4093 
                   
               
               
                   
                  5 
                 Slide36.jpg 
                 189 
                 1.34 
               
               
                   
                  6 
                 Slide36.jpg 
                 3572 
                 25.31 
               
               
                   
                  7 
                 Slide36.jpg 
                 4330 
                   
               
               
                   
                  8 
                 Slide36.jpg 
                 829 
                 9.38 
               
               
                   
                  9 
                 Slide36.jpg 
                 2710 
                 30.67 
               
               
                   
                 10 
                 Slide36.jpg 
                 2189 
                   
               
               
                   
                 11 
                 Slide36.jpg 
                 185 
                 1.18 
               
               
                   
                 12 
                 Slide36.jpg 
                 1581 
                 10.11 
               
               
                   
                 13 
                 Slide36.jpg 
                 1961 
                   
               
               
                   
                 14 
                 Slide36.jpg 
                 344 
                 1.40 
               
               
                   
                 15 
                 Slide36.jpg 
                 1293 
                 5.27 
               
               
                   
                 16 
                 Slide36.jpg 
                 2188 
                   
               
               
                   
                 17 
                 Slide36.jpg 
                 1766 
                 10.49 
               
               
                   
                 18 
                 Slide36.jpg 
                 382 
                 2.27 
               
               
                   
                 19 
                 Slide36.jpg 
                 4243 
                   
               
               
                   
                 20 
                 Slide36.jpg 
                 2206 
                 15.08 
               
               
                   
                 21 
                 Slide36.jpg 
                 1246 
                 8.52 
               
               
                   
                 22 
                 Slide36.jpg 
                 4883 
                   
               
               
                   
                 23 
                 Slide36.jpg 
                 3763 
                 37.76 
               
               
                   
                 24 
                 Slide36.jpg 
                 583 
                 5.85 
               
               
                   
                 25 
                 Slide36.jpg 
                 2162 
                   
               
               
                   
                 26 
                 Slide36.jpg 
                 2025 
                 13.11 
               
               
                   
                 27 
                 Slide36.jpg 
                 18 
                 0.12 
               
               
                   
                 28 
                 Slide36.jpg 
                 2558 
                   
               
               
                   
                 29 
                 Slide36.jpg 
                 1179 
                 3.69 
               
               
                   
                 30 
                 Slide36.jpg 
                 615 
                 1.92 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 54 
               
               
                   
               
               
                 TNF10 summary 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 non-IS 
                 46.76 
               
               
                   
                 IS 
                 51.20 
               
               
                   
                 LV 
                 97.96 
               
               
                   
                 IS/LV 
                 52% 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 55 
               
             
            
               
                   
               
               
                 Composite image data 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   
                 IS/LV 
                   
                 IS/LV 
                   
                 IS/LV 
                   
                 IS/LV 
               
               
                   
               
               
                 CON7 
                 70% 
                 TMZ3 
                 64% 
                 TNF1 
                 57% 
                 TNS1 
                 55% 
               
               
                 CONS 
                 65% 
                 TMZ1 
                 68% 
                 TNF2 
                 67% 
                 TNS2 
                 61% 
               
               
                 CON6 
                 75% 
                 TMZ2 
                 60% 
                 TNF3 
                 60% 
                 TNS3 
                 63% 
               
               
                 CON4 
                 65% 
                 TMZ7 
                 43% 
                 TNF4 
                 75% 
                 TNS4 
                 51% 
               
               
                 CON3 
                 64% 
                 TMZ8 
                 51% 
                 TNF5 
                 73% 
                 TNS5 
                 35% 
               
               
                 CON1 
                 77% 
                 TMZ5 
                 58% 
                 TNF6 
                 79% 
                 TNS6 
                 36% 
               
               
                 CON2 
                 55% 
                 TMZ6 
                 49% 
                 TNF7 
                 53% 
                 TNS7 
                 34% 
               
               
                 CON8 
                 68% 
                 TMZ9 
                 44% 
                 TNF8 
                 57% 
                 TNS8 
                 51% 
               
               
                 CON9 
                 67% 
                 TMZ10 
                 49% 
                 TNF9 
                 64% 
                 TNS9 
                 31% 
               
               
                 CON10 
                 62% 
                 TMZ4 
                 71% 
                 TNF10 
                 52% 
                 TNS10 
                 26% 
               
               
                 CON11 
                 73% 
                   
                   
                   
                   
                   
                   
               
               
                 CON12 
                 75% 
                   
                   
                   
                   
                   
                   
               
               
                 CON13 
                 76% 
                   
                   
                   
                   
                   
                   
               
               
                 CON14 
                 66% 
                   
                   
                   
                   
                   
                   
               
               
                 Mean 
                 68% 
                 Mean 
                 56% 
                 Mean 
                 64% 
                 Mean 
                 44% 
               
               
                 SD 
                  6% 
                 SD 
                 10% 
                 SD 
                 10% 
                 SD 
                 13% 
               
               
                 SE 
                  2% 
                 SE 
                  3% 
                 SE 
                  3% 
                 SE 
                  4% 
               
               
                 TTEST 
                   
                   
                 8.77E−04 
                   
                 1.61E−01 
                   
                 4.79E−06 
               
               
                   
                   
                   
                   
                   
                   
                 TMZ/TNS 
                 4.00E−02 
               
               
                   
               
            
           
         
       
     
     The results show that a combination of trimetazidine, nicotinamide, and succinate at 20 μM preserved coronary flow and cardiac functional recovery and decreased infarct size in isolated hearts after ischemia-reperfusion. This combination was more effective in decreasing infarct size than TMZ alone. A combination of trimetazidine, nicotinamide, and succinate at 2 μM did not appear to decrease myocardial ischemia-reperfusion injury. 
     This study suggested that the combination of trimetazidine, nicotinamide, and succinate at 20 μM generated better protection against ischemia-reperfusion injury in Langendorff system. 
       FIG. 43  is a schematic of the method used to analyze the effects of selected compositions on cardiac function. Following transverse aortic constriction (TAC) or a sham procedure, mice were given one of the following via an osmotic mini-pump: CV8814 at 5.85 mg/kg/day (CV4); CV8814 at 5.85 mg/kg/day, nicotinic acid at 1.85 mg/kg/day, and succinate at 2.43 mg/kg/day (TV8); or saline (SA). Echocardiograms were measured immediately following TAC, three weeks after TAC, and 6 weeks after TAC. Mice were sacrificed at 6 weeks, and tissues were analyzed. 
       FIG. 44  shows hearts from mice six weeks after a sham procedure (SHAM), TAC followed by saline administration (TAC), TAC followed by CV4 administration (CV4), or TAC followed by TV8 administration. 
       FIG. 45  is of graph of heart weight relative to body weight six weeks after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 46  is graph of heart weight six weeks after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 47  shows graphs of fractional shortening (FS) and ejection fraction (EF) at indicated time points after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 48  is a graph of left ventricular end-systolic diameter at indicated time points after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 49  is a graph of intraventricular septal dimension at indicated time points after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 50  is a graph of left ventricular mass at indicated time points after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 51  is a graph of isovolumic relaxation time at indicated time points after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 52  is a graph of the ratio peak velocity flow in early diastole vs. late diastole at indicated time points after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 53  is a graph of left ventricular developed pressure at six weeks after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
       FIG. 54  is a graph of the rate of left ventricle pressure rise at six weeks after transverse aortic constriction. Treatments are as indicated in relation to  FIG. 44 . 
     Chemical Synthesis Schemes 
     Compounds that shift cellular metabolism from fatty acid oxidation to glucose oxidation include 2-(4-(2,3,4-trimethoxybenzyl)piperazin-1-yl)ethan-1-ol (referred to herein as CV8814) and 2-(4-(2,3,4-trimethoxybenzyl)piperazin-1-yl)ethyl nicotinate (referred to herein as CV-8972). These compounds may be synthesized according to the following scheme: 
     Stage 1 
     
       
         
         
             
             
         
       
     
     Stage 2 
     
       
         
         
             
             
         
       
     
     Stage 3 
     
       
         
         
             
             
         
       
     
     The product was converted to the desired polymorph by recrystallization. The percentage of water and the ratio of methanol:methyl ethyl ketone (MEK) were varied in different batches using 2.5 g of product. 
     In batch MBA 25, 5% water w/r/t total volume of solvent (23 volumes) containing 30% methano1:70% MEK was used for precipitation. The yield was 67% of monohydrate of CV-8972. Water content was determined by KF to be 3.46%. 
     In batch MBA 26, 1.33% water w/r/t total volume of solvent (30 volumes) containing 20% methano1:80% MEK was used for precipitation. The yield was 86.5% of monohydrate of CV-8972. Water content was determined by KF to be 4.0%. The product was dried under vacuum at 40° C. for 24 hours to decrease water content to 3.75%. 
     In batch MBA 27, 3% water w/r/t total volume of solvent (32 volumes) containing 22% methano1:78% MEK was used for precipitation. The yield was 87.22% of monohydrate of CV-8972. Water content was determined by KF to be 3.93% after 18 hours of drying at room temperature under vacuum. The product was further dried under vacuum at 40° C. for 24 hours to decrease water content to 3.54%. 
     In other batches, the ratio and total volume of solvent were held constant at 20% methano1:80% MEK and 30 volumes in batches using 2.5 g of product, and only the percentage of water was varied. 
     In batch MBA 29, 1.0 equivalent of water was added. Material was isolated and dried under vacuum at 40° C. for 24 hours. Water content was determined by KF to be 0.89%, showing that the monohydrate form was not forming stoichiometrically. 
     In batch MBA 30, 3% water was added. Material was isolated and dried under vacuum at 40° C. for 24 hours. Water content was determined by KF to be 3.51%, showing that monohydrate is forming with addition of excess water. 
     In batch MBA 31, 5% water was added. Material was isolated and dried under vacuum at 40° C. for 24 hours. Water content was determined by KF to be 3.30%, showing that monohydrate is forming with addition of excess water. 
     Results are summarized in Table 56. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 56 
               
               
                   
               
               
                   
                 Water 
                   
                   
                   
                   
                   
                   
                   
                   
               
               
                   
                 percentage 
                   
                 KF result 
                 Amount of 
                   
                   
                   
                   
                   
               
               
                   
                 theoretical 
                 KF  
                 (Sample 
                 Water used 
                   
                   
                   
                   
                   
               
               
                   
                 (for 
                 result 
                 after drying 
                 for reaction 
                   
                   
                 Yield 
                 Drying 
                 Drying 
               
               
                   
                 monohydrate 
                 (% of 
                 at 40° C. for 
                 (based on total 
                 Ratio of 
                 Total 
                 obtained 
                 Time 
                 temperature 
               
               
                 Sample 
                 preparation) 
                 water) 
                 24 hours) 
                 volume) 
                 MeOH:MEK 
                 Volume 
                 (%) 
                 (hr) 
                 (° C.) 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 289-MBA-25 
                 3.32% 
                 3.46 
                 — 
                   5% 
                 30-70 
                 23 vol 
                 67.6 
                 24 
                 22 
               
               
                 289-MBA-26 
                 3.32% 
                 4.00 
                 3.75 
                 1.33% 
                 20-80 
                 30 vol 
                 86.5 
                 19 
                 23 
               
               
                 289-MBA-27 
                 3.32% 
                 3.93 
                 3.54 
                   3% 
                 22-78 
                 32 vol 
                 87.22 
                 18 
                 23 
               
               
                 289-MBA-29 
                 3.32% 
                 — 
                 0.89 
                 1.0 eq based 
                 20-80 
                 30 vol 
                 84 
                 24 
                 40 
               
               
                   
                   
                   
                   
                 on input 
                   
                   
                   
                   
                   
               
               
                   
                   
                   
                   
                 weight 
                   
                   
                   
                   
                   
               
               
                 289-MBA-30 
                 3.32% 
                 — 
                 3.51 
                   3% 
                 20-80 
                 30 vol 
                 90 
                 24 
                 40 
               
               
                 289-MBA-31 
                 3.32% 
                 — 
                 3.30 
                   5% 
                 20-80 
                 30 vol 
                 81 
                 24 
                 40 
               
               
                   
               
            
           
         
       
     
     Metabolism of Compounds in Dogs 
     The metabolism of various compounds was analyzed in dogs. 
       FIG. 55  is a graph showing levels of CV-8814 (solid triangles, solid lines) and trimetazidine (open triangles, dashed lines) after intravenous administration of CV-8834 at 2.34 mg/kg. CV-8834 is a compound of formula (II) in which y=1. 
       FIG. 56  is a graph showing levels of CV-8814 (solid triangles, solid lines) and trimetazidine (open triangles, dashed lines) after oral administration of CV-8834 at 77.4 mg/kg. 
       FIG. 57  is a graph showing levels of CV-8814 (solid triangles, solid lines) and trimetazidine (open triangles, dashed lines) after oral administration of CV-8834 at 0.54 mg/kg. 
       FIG. 58  is a graph showing levels of CV-8814 (solid triangles, solid lines) and trimetazidine (open triangles, dashed lines) after oral administration of CV-8834 at 1.08 mg/kg. 
       FIG. 59  is a graph showing levels of CV-8814 (solid triangles, solid lines) and trimetazidine (open triangles, dashed lines) after oral administration of CV-8834 at 2.15 mg/kg. 
     Data from  FIGS. 55-59  is summarized in Table 57. 
     
       
         
           
               
               
               
               
               
               
               
               
             
               
                 TABLE 57 
               
               
                   
               
               
                   
                 Route of 
                 Dose 
                   
                 T max   
                 C max   
                 AUC 0-8   
                   
               
               
                 Compound 
                 admin. 
                 (mg/kg) 
                 Analyte 
                 (hours) 
                 (ng/mL) 
                 (ng × hr/mL) 
                 % F 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 CV-8834 
                 PO 
                 77.4 
                 8814 
                 0.75 
                 12100 
                 38050 
                 69 
               
               
                 CV-8834 
                 PO 
                 77.4 
                 TMZ 
                 1.67 
                 288 
                 1600 
                 72 
               
               
                 CV-8834 
                 IV 
                 2.34 
                 8814 
                 0.083 
                 974 
                 1668 
                 — 
               
               
                 CV-8834 
                 IV 
                 2.34 
                 TMZ 
                 2.67 
                 13.4 
                 66.7 
                 — 
               
               
                 CV-8834 
                 PO 
                 0.54 
                 8814 
                 0.5 
                 74.0 
                 175 
                 45 
               
               
                 CV-8834 
                 PO 
                 0.54 
                 TMZ 
                 1.17 
                 3.63 
                 17.6 
                 &gt;100 
               
               
                 CV-8834 
                 PO 
                 1.08 
                 8814 
                 0.5 
                 136 
                 335 
                 44 
               
               
                 CV-8834 
                 PO 
                 1.08 
                 TMZ 
                 0.866 
                 6.19 
                 30.4 
                 99 
               
               
                 CV-8834 
                 PO 
                 2.15 
                 8814 
                 0.583 
                 199 
                 536 
                 35 
               
               
                 CV-8834 
                 PO 
                 2.15 
                 TMZ 
                 1.17 
                 9.80 
                 51.6 
                 84 
               
               
                   
               
            
           
         
       
     
       FIG. 60  is a graph showing levels of trimetazidine after oral administration of CV-8972 at 1.5 mg/kg (triangles) or intravenous administration of trimetazidine at 2 mg/kg (squares). 
       FIG. 61  is a graph showing levels of CV-8814 after oral administration of CV-8972 at 1.5 mg/kg (triangles) or intravenous administration of CV-8814 at 2.34 mg/kg (squares). 
       FIG. 62  is a graph showing levels of CV-8814 after intravenous administration of CV-8834 at 4.3 mg/kg (squares) or oral administration of CV-8834 at 2.15 mg/kg (triangles). 
       FIG. 63  is a graph showing levels of CV-8814 after intravenous administration of CV-8814 at 2.34 mg/kg (squares) or oral administration of CV-8814 at 2.34 mg/kg (triangles). 
     Data from  FIGS. 60-63  is summarized in Table 58. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 58 
               
               
                   
               
               
                   
                 Route of 
                 Dose 
                   
                   
                   
                 T max   
                 C max   
                 AUC 0-24   
                   
               
               
                 Compound 
                 admin. 
                 (mg/kg) 
                 Vehicle 
                 Fasted 
                 Analyte 
                 (hours)  
                 (ng/mL) 
                 (ng × hr/mL) 
                 % F 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
               
            
               
                 CV-8972 
                 PO 
                 1.5 
                 — 
                 — 
                 TMZ 
                 2.0 
                 17.0 
                 117 
                 4.3% 
               
               
                 TMZ 
                 IV 
                 2 
                 0.9% NaCl 
                 8 hrs 
                 TMZ 
                 0.083 
                 1002 
                 3612 
                 — 
               
               
                 CV-8972 
                 PO 
                 1.5 
                 — 
                 — 
                 8814 
                 1.125 
                 108 
                 534 
                  27% 
               
               
                 CV-8814 
                 IV 
                 2.34 
                 0.9% NaCl 
                 8 hrs 
                 8814 
                 0.083 
                 1200 
                 3059 
                 — 
               
               
                 CV-8834 
                 PO 
                 4.3 
                 0.9% NaCl 
                 8 hrs 
                 8814 
                 1.0 
                 692 
                 2871 
                  69% 
               
               
                 CV-8834 
                 IV 
                 4.3 
                 0.9% NaCl 
                 8 hrs 
                 8814 
                 0.083 
                 1333 
                 4154 
                 — 
               
               
                 CV-8834 
                 PO 
                 4.3 
                 0.9% NaCl 
                 8 hrs 
                 8814 
                 1.0 
                 692 
                 2871 
                  51% 
               
               
                 CV-8814 
                 IV 
                 2.34 
                 0.9% NaCl 
                 8 hrs 
                 8814 
                 0.083 
                 1200 
                 3059 
                 — 
               
               
                 CV-8814 
                 PO 
                 2.34 
                 0.9% NaCl 
                 8 hrs 
                 8814 
                 0.333 
                 672 
                 1919 
                  63% 
               
               
                 CV-8814 
                 IV 
                 2.34 
                 0.9% NaCl 
                 8 hrs 
                 8814 
                 0.083 
                 1200 
                 3059 
                 — 
               
               
                   
               
            
           
         
       
     
     Effect of CV-8814 on Enzyme Activity 
     The effect of CV-8814 on the activity of various enzymes was analyzed in in vitro assays. Enzyme activity was assayed in the presence of 10 μM CV-8814 using conditions of time, temperature, substrate, and buffer that were optimized for each enzyme based on published literature. Inhibition of 50% or greater was not observed for any of the following enzymes: ATPase, Na + /K + , pig heart; Cholinesterase, Acetyl, ACES, human; Cyclooxygenase COX-1, human; Cyclooxygenase COX-2, human; Monoamine Oxidase MAO-A, human; Monoamine Oxidase MAO-B, human; Peptidase, Angiotensin Converting Enzyme, rabbit; Peptidase, CTSG (Cathepsin G), human; Phosphodiesterase PDE3, human; Phosphodiesterase PDE4, human; Protein Serine/Threonine Kinase, PKC, Non-selective, rat; Protein Tyrosine Kinase, Insulin Receptor, human; Protein Tyrosine Kinase, LCK, human; Adenosine A1, human; Adenosine A 2A , human; Adrenergic α 1A , rat; Adrenergic α 1B , rat; Adrenergic α 1D , human; Adrenergic α 2A , human; Adrenergic α 2B , human; Adrenergic β 1 , human; Adrenergic β 2 , human; Androgen (Testosterone), human; Angiotensin AT 1 , human; Bradykinin B 2 , human; Calcium Channel L-Type, Benzothiazepine, rat; Calcium Channel L-Type, Dihydropyridine, rat; Calcium Channel L-Type, Phenylalkylamine, rat; Calcium Channel N-Type, rat; Cannabinoid CB 1 , human; Cannabinoid CB 2 , human; Chemokine CCR1, human; Chemokine CXCR2 (IL-8R B ), human; Cholecystokinin CCK 1  (CCK A ), human; Cholecystokinin CCK 2  (CCK B ), human; Dopamine D 1 , human; Dopamine D 2L , human; Dopamine D 2S , human; Endothelin ET A , human; Estrogen ERα, human; GABA A , Chloride Channel, TBOB, rat; GABA A , Flunitrazepam, Central, rat; GABA A , Ro-15-1788, Hippocampus, rat; GABA B1A , human; Glucocorticoid, human; Glutamate, AMPA, rat; Glutamate, Kainate, rat; Glutamate, Metabotropic, mGlu5, human; Glutamate, NMDA, Agonism, rat; Glutamate, NMDA, Glycine, rat; Glutamate, NMDA, Phencyclidine, rat; Glutamate, NMDA, Polyamine, rat; Glycine, Strychnine-Sensitive, rat; Histamine H 1 , human; Histamine H 2 , human; Melanocortin MC 1 , human; Melanocortin MC 4 , human; Muscarinic M 1 , human; Muscarinic M 2 , human; Muscarinic M 3 , human; Muscarinic M 4 , human; Neuropeptide Y Y 1 , human; Nicotinic Acetylcholine, human; Nicotinic Acetylcholine α1, Bungarotoxin, human; Opiate δ 1  (OP1, DOP), human; Opiate κ (OP2, KOP), human; Opiate μ (O P3, MOP), human; Platelet Activating Factor (PAF), human; Potassium Channel [KATP], hamster; Potassium Channel hERG, human; PPARγ, human; Progesterone PR-B, human; Serotonin (5-Hydroxytryptamine) 5-HT 1A , human; Serotonin (5-Hydroxytryptamine) 5-HT 1B , human; Serotonin (5-Hydroxytryptamine) 5-HT 2A , human; Serotonin (5-Hydroxytryptamine) 5-HT 2B , human; Serotonin (5-Hydroxytryptamine) 5-HT 2C , human; Serotonin (5-Hydroxytryptamine) 5-HT 3 , human; Sodium Channel, Site 2, rat; Tachykinin NK 1 , human; Transporter, Adenosine, guinea pig; Transporter, Dopamine (DAT), human; Transporter, GABA, rat; Transporter, Norepinephrine (NET), human; Transporter, Serotonin (5-Hydroxytryptamine) (SERT), human; and Vasopressin V 1A , human. 
     Analysis of CV-8972 Batch Properties 
     CV-8972 (2-(4-(2,3,4-trimethoxybenzyl)piperazin-1-yl)ethyl nicotinate, HCl salt, monohydrate) was prepared and analyzed. The batch was determined to be 99.62% pure by HPLC. 
       FIG. 64  is a graph showing the HPLC elution profile of a batch of CV-8972. 
       FIG. 65  is a graph showing analysis of molecular species present in a batch of CV-8972. 
       FIG. 66  is a pair of graphs showing HPLC elution profiles of molecular species present in a batch of CV-8972. 
       FIG. 67  is a pair of graphs showing HPLC elution profiles of molecular species present in a batch of CV-8972. 
       FIG. 68  is a graph showing X-ray powder diffraction analysis of a batch of CV-8972. 
       FIG. 69  is a graph showing X-ray powder diffraction analysis of batches of CV-8972. Batch 289-MBA-15-A, shown in blue, contains form B of CV-8972, batch 276-MBA-172, shown in black contains form A of CV-8972, and batch 289-MBA-16, shown in red, contains a mixture of forms A and B. 
       FIG. 70  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of batch 276-MBA-172 of CV-8972. 
       FIG. 71  is a graph showing dynamic vapor sorption (DVS) of batch 276-MBA-172 of CV-8972. 
       FIG. 72  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of batch 289-MBA-15-A of CV-8972. 
       FIG. 73  is a graph showing dynamic vapor sorption (DVS) of batch 289-MBA-15-A of CV-8972. 
       FIG. 74  is a graph showing X-ray powder diffraction analysis of samples of CV-8972. A pre-DVS sample from batch 276-MBA-172 is shown in blue, a pre-DVS sample from batch 289-MBA-15-A is shown in red, and a post-DVS sample from batch 289-MBA-15-A is shown in black. 
       FIG. 75  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of batch 289-MBA-16 of CV-8972. 
       FIG. 76  is a graph showing X-ray powder diffraction analysis of samples of CV-8972. Form B is shown in green, form A is shown in blue, a sample from an ethanol slurry of batch 289-MBA-15-A is shown in red, and a sample from an ethanol slurry of batch 289-MBA-16 is shown in black. 
     The stability of CV-8972 was analyzed. 
     Samples from batch 289-MBA-15-A (containing form B) were added to various solvents, incubated under various conditions, and analyzed by X-ray powder diffraction. Results are summarized in Table 59. 
     
       
         
           
               
               
               
             
               
                 TABLE 59 
               
               
                   
               
               
                 Solvent 
                 Conditions 
                 XRPD results 
               
               
                   
               
             
            
               
                 EtOH 
                 Slurry, RT, 3 d 
                 Form A + Form B 
               
               
                 MeOH/H2O (95:5) A w  = 0.16 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 IPA/H2O (98:2) A w  = 0.26 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 MeOH/H2O (80:20) A w  = 0.48 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 EtOH/H2O (90:10) A w  = 0.52 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 IPA/H2O (90:10) A w  = 0.67 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 Acetone/H2O (90:10) A w  = 0.72 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 ACN/H2O (90:10) A w  = 0.83 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 EtOAc/H2O (97:3) A w  = 0.94) 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 MeOH 
                 Slurry, RT, 5 d 
                 Form A + Form B 
               
               
                 EtOAc 
                 Slurry, RT, 5 d 
                 Form A + Form B 
               
               
                 MEK 
                 Slurry, RT, 5 d 
                 Form A 
               
               
                 — 
                 100° C., 
                 Form B, shifted with  
               
               
                   
                 20 minutes 
                 minor Form A 
               
               
                 EtOH 
                 CC from 60° C. 
                 Form C + minor  
               
               
                   
                   
                 Form A 
               
               
                   
               
            
           
         
       
     
     Samples from batch 289-MBA-16 (containing forms A and B) were added to various solvents, incubated under various conditions, and analyzed by X-ray powder diffraction. Results are summarized in Table 60. 
     
       
         
           
               
               
               
               
             
               
                   
                 TABLE 60 
               
               
                   
                   
               
               
                   
                 Solvent 
                 Conditions 
                 XRPD results 
               
               
                   
                   
               
             
            
               
                   
                 EtOH 
                 Slurry, RT, 3 d 
                 Form A + Form B 
               
               
                   
                 MeOH 
                 Vapor diffusion w/MTBE 
                 Form A 
               
               
                   
                 EtOAc 
                 Attempted to dissolve at ~60° C.,  
                 Form A + Form B 
               
               
                   
                   
                 solids remained, cooled slowly 
                   
               
               
                   
                   
                 to RT, let stir at RT from 60° C. 
               
               
                   
                   
               
            
           
         
       
     
       FIG. 77  is a graph showing X-ray powder diffraction analysis of samples of CV-8972. A sample containing form B is shown in blue, a sample containing form A is shown in red, and a sample containing a mixture of forms A and C is shown in black. 
     The stability of CV-8972 was analyzed. Aqueous samples containing CV-8972 at different concentrations and pH were incubated for various periods and analyzed. Results are shown in Table 61. 
     
       
         
           
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 61 
               
             
            
               
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 Decrease 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 in purity 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 of CV-8972 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 between  
               
            
           
           
               
               
               
               
               
            
               
                   
                 Time 
                   
                 Retention Time 
                 time 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 Sample 
                 (hrs) 
                 pH 
                 2.2 
                 2.6 
                 4.2 
                 4.7 
                 5.6 
                 points 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 276-MBA-172 
                 0 
                 6.6 
                 3.39 
                 0.6 
                 0.23 
                 0.54 
                 95.24 
                   
               
               
                 10 mg/mL pH 6 
                 1 
                 6.8 
                 4.81 
                 0.81 
                 0.23 
                 0.73 
                 93.43 
                 1.81 
               
               
                 (Form A) 
                 4 
                 6.8 
                 5.72 
                 0.9 
                 0.21 
                 0.83 
                 91.82 
                 1.61 
               
               
                   
                 6 
                 6.7 
                 6.45 
                 0.81 
                 ND 
                 0.93 
                 91.8 
                 0.02 
               
               
                   
                 22 
                 6.7 
                 7.38 
                 1.54 
                 0.13 
                 1.11 
                 89.66 
                 2.14 
               
               
                 276-MBA-172 
                 0 
                 6.1 
                 ND 
                 ND 
                 1.29 
                 ND 
                 98.01 
                   
               
               
                 2 mg/mL pH 6 
                 1 
                 6.1 
                 1.5 
                 ND 
                 1.28 
                 ND 
                 97.22 
                 0.79 
               
               
                 (Form A) 
                 4 
                 6.1 
                 2.03 
                 ND 
                 0.95 
                 ND 
                 97.01 
                 0.21 
               
               
                   
                 6 
                 6.1 
                 2.47 
                 ND 
                 1.02 
                 ND 
                 96.51 
                 0.5 
               
               
                   
                 22 
                 6.1 
                   
                   
                   
                   
                   
                   
               
               
                 289-MBA-15-A 
                 10 
                 0 
                 6 
                 3.3 
                 0.6 
                 0.26 
                 0.48 
                 95.36 
               
               
                 mg/mL pH 6  
                 1 
                 6.1 
                 3.76 
                 0.65 
                 0.25 
                 0.53 
                 94.81 
                 0.55 
               
               
                 (Form B) 
                 4 
                 6 
                 3.97 
                 0.59 
                 0.19 
                 0.56 
                 94.69 
                 0.12 
               
               
                   
                 6 
                 5.9 
                 4.3 
                 0.54 
                 0.17 
                 0.6 
                 94.39 
                 0.3 
               
               
                   
                 22 
                 5.9 
                 4.53 
                 0.69 
                 0.19 
                 0.65 
                 93.93 
                 0.46 
               
               
                 289-MBA-15-A 
                 2 
                 0 
                 6.9 
                 1.33 
                 ND 
                 1.19 
                 ND 
                 97.48 
               
               
                 mg/mL pH 6  
                 1 
                 6.9 
                 3.73 
                 ND 
                 1.17 
                 ND 
                 95.1 
                 2.38 
               
               
                 (Form B) 
                 4 
                 6.8 
                 5.25 
                 0.67 
                 0.84 
                 0.79 
                 92.45 
                 2.65 
               
               
                   
                 6 
                 6.8 
                 6.63 
                 0.9 
                 0.83 
                 0.99 
                 90.65 
                 1.8 
               
               
                   
                 22 
                 6.7 
                 7.72 
                 1.13 
                 0.86 
                 1.14 
                 89.15 
                 1.5 
               
               
                 276-MBA-172 
                 10 
                 0 
                 7.1 
                 5.9 
                 0.94 
                 0.22 
                 0.78 
                 92.85 
               
               
                 mg/mL pH 7  
                 1 
                 7.2 
                 8.12 
                 1.45 
                 0.21 
                 1.17 
                 89.05 
                 3.8 
               
               
                 (Form A) 
                 4 
                 7.1 
                 10.14 
                 1.48 
                 0.13 
                 1.46 
                 86.8 
                 2.25 
               
               
                   
                 6 
                 7.1 
                 11.63 
                 1.78 
                 0.13 
                 1.67 
                 84.79 
                 2.01 
               
               
                   
                 22 
                 7 
                   
                   
                   
                   
                   
                   
               
               
                 276-MBA-172 
                 2 
                 0 
                 6.7 
                 1.42 
                 ND 
                 1.05 
                 ND 
                 97.53 
               
               
                 mg/mL pH 7  
                 1 
                 6.8 
                 3.31 
                 ND 
                 1.06 
                 0.57 
                 95.06 
                 2.47 
               
               
                 (Form A) 
                 4 
                 6.7 
                 4.21 
                 0.58 
                 0.82 
                 0.69 
                 93.7 
                 1.36 
               
               
                   
                 6 
                 6.7 
                 5.63 
                 0.67 
                 0.74 
                 0.85 
                 92.12 
                 1.58 
               
               
                   
                 22 
                 6.8 
                 6.26 
                 0.85 
                 0.85 
                 0.98 
                 91.07 
                 1.05 
               
               
                 289-MBA-15-A 
                 10 
                 0 
                 7.4 
                 6.2 
                 1.16 
                 0.27 
                 0.87 
                 91.5 
               
               
                 mg/mL pH 7  
                 1 
                 7.4 
                 10.47 
                 1.65 
                 0.25 
                 1.44 
                 86.18 
                 5.32 
               
               
                 (Form B) 
                 4 
                 7.4 
                 13.64 
                 1.93 
                 0.19 
                 1.89 
                 82.36 
                 3.82 
               
               
                   
                 6 
                 7.3 
                 15.66 
                 2.57 
                 0.2 
                 0.2 
                 79.37 
                 2.99 
               
               
                   
                 22 
                 7.1 
                   
                   
                   
                   
                   
                   
               
               
                 289-MBA-15-A 
                 2 
                 0 
                 6.5 
                 1.62 
                 ND 
                 0.9 
                 ND 
                 97.48 
               
               
                 mg/mL pH 7  
                 1 
                 6.6 
                 3.16 
                 ND 
                 0.89 
                 0.49 
                 95.46 
                 2.02 
               
               
                 (Form B) 
                 4 
                 6.5 
                 4.27 
                 0.53 
                 0.66 
                 0.62 
                 93.92 
                 1.54 
               
               
                   
                 6 
                 6.5 
                   
                   
                   
                   
                   
                   
               
               
                   
                 22 
                 6.5 
               
               
                   
               
            
           
         
       
     
     Samples from batch S-18-0030513 (containing form A) were added to various solvents, incubated under various conditions, and analyzed by X-ray powder diffraction. Results are summarized in Table 62. 
     
       
         
           
               
               
               
             
               
                 TABLE 62 
               
               
                   
               
               
                 Solvent 
                 Conditions 
                 XRPD results 
               
               
                   
               
             
            
               
                 CHCl3 
                 Slurry, RT 
                 Form A 
               
               
                 EtOAc 
                 Slurry, RT 
                 Form A 
               
               
                 THF 
                 Slurry, RT 
                 Form A 
               
               
                 — 
                 VO, RT 
                 Form A 
               
               
                 — 
                  80° C, 20 minutes 
                 Form A with slight peak shifting 
               
               
                 — 
                 100° C, 20 minutes 
                 Form B + Form A, shifted 
               
               
                 — 
                 97% RH Stress of 
                 Form A 
               
               
                   
                 Form A dried at 
                   
               
               
                   
                 80° C. for 20 min 
                   
               
               
                 EtOH 
                 Crash cool from 
                 Form A + Form C 
               
               
                   
                 70° C. 
                   
               
               
                 MEK/H2O 99:1 
                 Slow cool from 
                 Form A 
               
               
                   
                 70° C. 
               
               
                   
               
            
           
         
       
     
     Samples from batch 289-MBA-16 (containing forms A and B) were added to various solvents, incubated under various conditions, and analyzed by X-ray powder diffraction. Results are summarized in Table 63. 
     
       
         
           
               
               
               
               
             
               
                   
                 TABLE 63 
               
               
                   
                   
               
               
                   
                 Solvent 
                 Conditions 
                 XRPD results 
               
               
                   
                   
               
             
            
               
                   
                 EtOH 
                 Slurry, RT, 3 d 
                 Form A + Form B 
               
               
                   
                 MeOH 
                 VD w/MTBE 
                 Form A 
               
               
                   
                 EtOAc 
                 SC from 60° C. 
                 Form A + Form B 
               
               
                   
                 THF 
                 SC from 60° C. 
                 Form B 
               
               
                   
                 EtOH 
                 SC from 60° C. 
                 Form A + Form C 
               
               
                   
                 MeOH/H2O  
                 Slurry, overnight,  
                 Form A 
               
               
                   
                 (95:5) 
                 1 g scale 
               
               
                   
                   
               
            
           
         
       
     
       FIG. 78  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of samples containing form A of CV-8972. A sample from an ethanol acetate-water slurry is shown with solid lines, a sample from a methanol-water slurry is shown with regularly-dashed lines, and a sample from an ethanol-water slurry is shown with dashed-dotted lines. 
       FIG. 79  is a graph showing differential scanning calorimetry and thermal gravimetric analysis of a sample containing form A of CV-8972. Prior to analysis, the sample was dried at 100° C. for 20 minutes. 
     Samples containing form A of CV-8972 were analyzed for stability in response to humidity. Samples were incubated at 40 ° C., 75% relative humidity for various periods and analyzed. Results are shown in Table 64. 
     
       
         
           
               
               
             
               
                 TABLE 64 
               
             
            
               
                   
               
               
                 Time  
                 Retention Time 
               
            
           
           
               
               
               
               
               
            
               
                 (days) 
                 1.9 
                 3.9 
                 4.5 
                 5.4 
               
               
                   
               
               
                 0 
                 ND 
                 1.16 
                 ND 
                 98.84 
               
               
                 1 
                 ND 
                 0.68 
                 ND 
                 99.32 
               
               
                 7 
                 0.63 
                 0.14 
                 0.12 
                 99.12 
               
               
                   
               
            
           
         
       
     
     Form A of CV-8972 was analyzed for stability in aqueous solution. Aqueous samples containing CV-8972 at different concentrations and pH were incubated for various periods and analyzed. Results are shown in Table 65. 
     
       
         
           
               
               
               
               
             
               
                 TABLE 65 
               
             
            
               
                   
               
               
                   
                   
                   
                 % change 
               
               
                 Concentration  
                 Time 
                 Retention Time 
                 from t0 of 
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                 of CV-8972 
                 (hrs) 
                 1.9 
                 2.2 
                 3.9 
                 4.5 
                 5.4 
                 RT 5.4 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
            
               
                   21 mg/mL,  
                 0 
                 ND 
                 ND 
                 1.12 
                 ND 
                 98.88 
                 — 
               
               
                 Initial 
                 1 
                 1.03 
                 ND 
                 0.94 
                 ND 
                 98.03 
                 −0.86 
               
               
                 pH = 2.0 
                 2 
                 1.9 
                 ND 
                 1 
                 ND 
                 97.11 
                 −1.79 
               
               
                   
                 6 
                 5.25 
                 0.83 
                 0.96 
                 0.78 
                 92.18 
                 −6.78 
               
               
                 12.5 mg/mL,  
                 0 
                 ND 
                 ND 
                 1.79 
                 ND 
                 98.21 
                 — 
               
               
                 Initial 
                 1 
                 1.38 
                 ND 
                 1.41 
                 ND 
                 97.21 
                 −1.02 
               
               
                 pH = 2.1 
                 2 
                 2.43 
                 ND 
                 1.67 
                 ND 
                 95.9 
                 −2.35 
               
               
                   
                 6 
                 6.59 
                 1.04 
                 1.74 
                 1.04 
                 89.58 
                 −8.79 
               
               
                  4.2 mg/mL,  
                 0 
                 ND 
                 ND 
                 5.35 
                 ND 
                 94.65 
                 — 
               
               
                 Initial 
                 1 
                 ND 
                 ND 
                 4.02 
                 ND 
                 95.98 
                 1.41 
               
               
                 pH = 2.3 
                 2 
                 3.72 
                 ND 
                 5.09 
                 ND 
                 91.19 
                 −3.66 
               
               
                   
                 6 
                 9.71 
                 ND 
                 5.3 
                 ND 
                 84.99 
                 −10.21 
               
               
                   
               
            
           
         
       
     
     The amount of CV-8972 present in various dosing compositions was analyzed. Results are shown in Table 66. 
     
       
         
           
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 66 
               
               
                   
               
               
                   
                   
                   
                   
                   
                 Total 
                 pH 
                   
                   
               
               
                   
                   
                   
                   
                   
                 vol.  
                 after 
                 Vol. addl. 
                   
               
               
                 Target 
                 Vol. API 
                 Mass 
                 Initial 
                 Vol. 1N 
                 base 
                 base  
                 1N NaOH 
                 Final  
               
               
                 Dose 
                 soln. 
                 CV8972 
                 pH API 
                 NaOH 
                 soln. 
                 soln. 
                 added  
                 Dose 
               
               
                 (mg/mL) 
                 (mL) 
                 (mg) 
                 soln. 
                 (mL) 
                 (mL) 
                 addn. 
                 (mL) 
                 (mg/mL) 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 10 
                 30 
                 779.06 
                 2.0 
                 2.07 
                 30 
                 3.6 
                 0.7 
                 9.92 
               
               
                 2 
                 30 
                 157.38 
                 2.4 
                 0.19 
                 30 
                 2.8 
                 0.35 
                 2.02 
               
               
                 10 
                 50 
                 777.05 
                 2.1 
                 2.77 
                 10 
                 6.2 
                 — 
                 10.01 
               
               
                 2 
                 50 
                 142.08 
                 2.5 
                 0.99 
                 10 
                 3.0 
                 0.3 
                 1.82 
               
               
                   
               
            
           
         
       
     
     Brain-To-Plasma Ratio of Compounds In Vivo 
     The brain-to-plasma ratio of trimetazidine and CV-8814 was analyzed after intravenous administration of the compounds to rats. Dosing solutions were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Results are shown in Table 67. 
     
       
         
           
               
               
               
               
               
               
             
               
                 TABLE 67 
               
               
                   
               
               
                   
                   
                   
                   
                 Measured 
                   
               
               
                   
                   
                   
                 Nominal 
                 Dosing 
                   
               
               
                   
                   
                   
                 Dosing  
                 Solution 
                   
               
               
                 Test  
                 Route of 
                   
                 Conc. 
                 Conc. 
                 % of  
               
               
                 Article 
                 Administration 
                 Vehicle 
                 (mg/mL) 
                 (mg/mL) 
                 Nominal 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 TMZ 
                 IV 
                 Normal 
                 1.0 
                 1.14 
                 114 
               
               
                   
                   
                 Saline* 
                   
                   
                   
               
               
                 CV-8814 
                 IV 
                 Normal 
                 0.585 
                 0.668 
                 114 
               
               
                   
                   
                 Saline* 
                   
                   
                   
               
               
                   
               
            
           
         
       
     
     The concentrations of compounds in the brain and plasma were analyzed 2 hours after administering compounds at 1 mg/kg to rats. Results from trimetazidine-treated rats are shown in Table 68. Results from CV-8814-treated rats are shown in Table 69. 
     
       
         
           
               
             
               
                 TABLE 68 
               
             
            
               
                   
               
               
                 TMZ-treated rats 
               
            
           
           
               
               
               
               
            
               
                 Rat# 
                 11 
                 12 
                 13 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Brain Weight (g) 
                 1.781 
                 1.775 
                 1.883 
               
               
                 Brain Homogenate Volume (mL) 
                 8.91 
                 8.88 
                 9.42 
               
               
                 Brain Homogenate Conc. (ng/mL) 
                 7.08 
                 7.35 
                 7.90 
               
               
                 Brain Tissue Conc. (ng/g) 
                 35.4 
                 36.8 
                 39.5 
               
               
                 Plasma Conc. (ng/g) 1   
                 22.7 
                 14.0 
                 14.1 
               
               
                 B:P Ratio 
                 1.56 
                 2.63 
                 2.80 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 69 
               
             
            
               
                   
               
               
                 CV-8814-treated rats 
               
            
           
           
               
               
               
               
               
            
               
                   
                 Rat# 
                 14 
                 15 
                 16 
               
               
                   
                   
               
            
           
           
               
               
               
               
               
            
               
                   
                 Brain Weight (g) 
                 1.857 
                 1.902 
                 2.026 
               
               
                   
                 Brain Homogenate Volume  
                 9.29 
                 9.51 
                 10.1 
               
               
                   
                 (mL) 
                   
                   
                   
               
               
                   
                 Brain Homogenate Conc.  
                 4.01 
                 4.21 
                 4.74 
               
               
                   
                 (ng/mL) 
                   
                   
                   
               
               
                   
                 Brain Tissue Conc. (ng/g) 
                 20.1 
                 21.1 
                 24 
               
               
                   
                 Plasma Conc. (ng/g) 1   
                 19.3 
                 17.0 
                 14.0 
               
               
                   
                 B:P Ratio 
                 1.04 
                 1.24 
                 1.693 
               
               
                   
                   
               
            
           
         
       
     
     The average B:P ratio for trimetazidine-treated rats was 2.33 ±0.672. The average B:P ratio for trimetazidine-treated rats was 1.32 ±0.335. 
     Incorporation by Reference 
     References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made throughout this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes. 
     Equivalents 
     Various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including references to the scientific and patent literature cited herein. The subject matter herein contains important information, exemplification, and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.