Omefibrates for Treating Dyslipidemia and Cardiovascular Disease

Abstract

The present invention relates to the fibric acid derivatives of omega-3 fatty acids and their use in treating Type2 diabetes, obesity, hypertriglyceridemia, cardiovascular diseases, metabolic syndrome, cancer, Alzheimer's disease; and their use for modulating activity of peroxisome proliferator-activated receptors (PPARs).

Claims

1. A method for treating hypertrilyceridemia, cardiovascular disease, metabolic syndrome, Type2 diabetes, obesity, cancer, renal anemia, or Alzheimer's disease in a arson in need of such treatment by administering an effective amount of a formulation wherein the active ingredient is a compound of Formula (I): ##STR00034## wherein: R.sub.1 is —H, —C.sub.2H.sub.5, —C.sub.3H.sub.7, —CH(CH.sub.3).sub.3, —C(CH.sub.3).sub.3, or —C(C.sub.2H.sub.5)CH.sub.3).sub.2 or when R.sub.1 is —H and when it is converted to its metformin salt, then R.sub.1 is a metformin cation of the formula ##STR00035## R is joined from the methylene moiety formed by reduction of the carboxylic acid of one of the following polyunsaturated fatty acids (PUFAs): cis,cis,cis-7,10,13-hexadecatrienoic acid (HTA), cis,cis,cis-9,12,15-octadecatrienoic acid (ALA), cis,cis,cis,cis-6,9,12,15-octadecatetraenoic acid (SDA), cis,cis,cis-11,14,17-eicosatrienoic acid (ETE), cis,cis,cis,cis-8,11,14,17-eicosatetraenoic acid (ETA); cis,cis,cis,cis-5,8,11,14,17-eicosapentanenoic acid (EPA), cis,cis,cis,cis-6,12,15,18-heneicosapentaenoic acid (HPA), cis,cis,cis,cis-7,10,13,16,19-docosapentaenoic acid (DPA), cis,cis,cis,cis,cis-4,7,10,13,16,19-docosahexaenoic acid (DHA), cis,cis,cis,cis,cis-9,12,15,18,21-tetracosapentaeonic acid (TPA) or cis,cis,cis,cis,cis,cis-6,9,12,15,18,21-tetracosahexaeonic acid (THA); including its pharmaceutically-acceptable salts, with adjuvant, binders, desiccants, diluents and excipients.

2. The method of claim 1 wherein R.sub.1 is the metformin salt of Formula (I) wherein R.sub.1 is —H; as shown by Formula (II): ##STR00036## wherein R is defined as in claim 1.

3. The method of claim 1 wherein the compound is ≧90% chemical purity.

4. The method of claim 1 having as its active ingredient one or more compounds of Formula (I) as defined in claim 1.

5. The method of claim 1 wherein the active ingredient is in a formulation in the form of a solution for injection, ampoule, hard or soft gelatin capsule or tablet, or as a sustained release formulation.

6. The method of claim 1 to treat hypertriglyceridemia, Type2 diabetes, or metabolic syndrome in persons needing such treatment by administering to such persons an effective amount of the active ingredient in the formulation.

7. The method of claim 6 to treat metabolic syndrome wherein R.sub.1 in Formula (I) is the metformin cation.

8. The method of claim 6, wherein the triglycerides levels in such persons needing treatment are in a range of from >100 mg/dl, to >500 mg/dl.

9. The method of claim 1 for the treatment of early stages of Alzheimer's disease in persons needing such treatment by administering to such person an effective amount of the active ingredient of the formulation.

10. The method of claim 6, wherein the effective amount is from about 0.05 to about 5 g/day administered as 1-4 doses/day.

11. (canceled)

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0019] FIG. 1 is a graph in color showing the results from testing of 3 compounds of the present invention (JIVA-0018 green dots. JIVA-0019 pink dots; JIVA-0020 orange dots) compared to 2 standards (bezafibrate red dots; GW7647 black dots) in a PPARα Agonist Assay where the normalized luciferase activity is plotted vs. the log of the concentration (M) of the tested compound. JIVA-0018 and bezafibrate are at the base line.

[0020] FIG. 2 is a graph in color showing the results from testing of 3 compounds of the present invention (JIVA-0018 green dots, JIVA-0019 pink dots; JIVA-0020 orange dots) compared to 1 standard (GW7647 red dots) in a PPARδ Agonist Assay where the normalized luciferase activity is plotted vs. the log of the concentration (M) of the tested compound. JIVA-0018 and JIVA-0019 are at the base line.

[0021] FIG. 3 is a graph in color showing the results from testing of 4 compounds of the present invention (JIVA-0013 blue dots. JIVA-0018 green dots, JIVA-0019 pink dots; JIVA-0020 orange dots) of the tested compound compared to 1 standard (bezafibrate red dots) in a PPARγ Agonist Assay where the normalized luciferase activity is plotted vs. the log of the concentration (M) of the tested compound.

DETAILED DESCRIPTION OF THE INVENTION

[0022] It is understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. As used in this specification, the singular forms “a”, “an”, and “the” include plural referents unless the content clearly indicates otherwise. The following terms in the Glossary as used in this application are to be defined as stated below and for these terms, the singular includes the plural.

[0023] Various headings are present to aid the reader, but are not the exclusive location of all aspects of that referenced subject matter and are not to be construed as limiting the location of such discussion.

[0024] Also, certain US patents and PCT published applications have been incorporated by reference. However, the text of such patents is only incorporated by reference to the extent that no conflict exists between such text and other statements set forth herein. In the event of such conflict, then any such conflicting text in such incorporated by reference US patent or PCT application is specifically not so incorporated in this patent.

Glossary

[0025] ALA means α-linolenic acid or cis,cis,cis-9,12,15-octadecatrienoic acid, having 18 carbons, 3 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (11Z,14Z,17Z)-eicosa-11,14,17-trien-2-yl)-2,2-dimethyleicosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00004##

DHA means cis,cis,cis,cis,cis,cis-4,7,10,13,16,19-docosahexaenoic acid or docosahexaenoic acid, having 22 carbons, 6 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (6Z,9Z, 12Z,15Z,18Z,21Z)tetracosa-69,12,15,18,21-hexaen-2-yl)-2,2-dimethyl-tetracosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00005##

DPA means cis,cis,cis,cis,cis-7,10,13,16,19-docosapentaenoic acid or docosapentaenoic acid, having 22 carbons, 5 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (9Z,12Z, 15Z,18Z,21Z)tetracosa-9,12,15,18,21-pentaen-2-yl)-2,2-dimethyltetracosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00006##

EPA means cis,cis,cis,cis,cis-5,8,11,14,17-eicosapentanenoic acid or eicosapentanenoic acid, having 20 carbons, 5 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (7Z,10Z,13Z,16Z,19Z)-docosa-7,10,13,16,19-pentaen-2-yl)-2,2-dimethyldocosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00007##

ETA means cis,cis,cis,cis-8,11,14,17-eicosatetranoic acid or eicosatetraenoic acid, having 20 carbons, 4 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (10Z,13Z,16Z,19Z)-docosa-10,13,16,19-tetraen-2-yl)-2,2-dimethyldocosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00008##

ETE means cis,cis,cis-11,14,17-eicosatrienoic acid or eicosatrienoic acid, having 20 carbons, 3 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (13Z,16Z,19Z)-docosa-13,16,19-triene-2-yl)-2,2-dimethyldocosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00009##

HPA means cis,cis,cis,cis,cis-6,9,12,15,18-heneicosapentaenoic acid or heneicosapentaenoic acid, having 21 carbons, 5 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (8Z 11Z,14Z, 17Z,20Z)-tricosa-8,11,14,17,20-pentaen-2-yl)-2,2-dimethyltricosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00010##

HTA means cis,cis,cis-7,10,13-hexadecatrienoic acid, having 16 carbons, 3 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (9Z,12 Z, 15Z)-octadeca-9,12,15-trien-2-yl)-2,2-dimethyloctadecanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00011##

SDA means cis,cis,cis,cis-6,9,12,15-octadecatetraenoic acid or stearidonic acid, having 18 carbons. 4 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (8Z,11Z,14Z,17Z)-eicosa-8,11,14,17-tetraen-2-yl)-2,2-dimethyleicosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00012##

THA means cis, cis, cis, cis,cis,cis-6,9,12,15,18,21-tetracosahexaeonic acid, having 24 carbons, 6 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (8Z,11 Z, 14Z, 17Z,20Z,23Z)-hexacosa-8,11,14,17,20,23-hexaen-2-yl)-2,2-dimethylhexacosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00013##

TPA means cis,cis,cis,cis,cis-9,12,15,18,21-tetracosapentaeonic acid, having 24 carbons, 5 cis double bonds, that is modified by reduction of the carboxylic acid to a methylene moiety to be R of Formula (I), (11Z,14Z,17Z,20Z,23Z)-hexacosa-11,14,17,20,23-pentaen-2-yl)-2,2-dimethylhexacosanoic acid, and its ethyl, n-propyl, isopropyl, t-butyl, and dimethylethyl esters, as shown by the formula below:

##STR00014##

Brine means a saturated solution of sodium chloride in water at room temperature (RT), typically 26% concentration w/v.
DMSO means dimethylsulfoxide
Omega-3 fatty acids means naturally occurring, straight-chain C.sub.16-C.sub.24 fatty carboxylic acids
PUFA means polyunsaturated fatty acids that are either naturally occurring omega-3 fatty acids or derivatives thereof.
DIBALH means diisobutylaluminumhydride
hr means hour(s)
LDA means lithium diisopropylamide
LAH means lithium aluminumhydride
min. means minute(s)
Omefibrates means fibric acid type derivatives of omega-3 polyunsaturated acids (fatty acids)
Ph means phenyl
RT means room temperature or ambient temperature or about 22 to about 25° C.
rt means retention time in the context of high performance liquid chromatography to determine purity of a compound
THF means tetrahydrofuran
TLC means thin layer chromatography
HRMS means high resolution mass spectroscopy
w/w means weight by weight
w/v means weight per volume

[0026] The present invention provides fibrate compounds of Formula (I) that are derived from the above polyunsaturated omega-3 fatty acids (PUFAs) as triglyceride reducing agents of the formula

##STR00015##

[0027] wherein: R.sub.1 is —H, —C.sub.2H.sub.5, —C.sub.3H.sub.7, —CH(CH.sub.3).sub.2, —C(CH.sub.3).sub.3 or —C(C.sub.2H.sub.5)(CH.sub.3).sub.2 or when R.sub.1 is —H and when it is converted to its metformin salt then R.sub.1 is a metformin cation of the formula

##STR00016##

Therefore when the metformin salt is formed from Formula (I) it is shown by Formula (II) and is used to treat metabolic syndrome.

##STR00017##

[0028] wherein: in both Formula (I) and (II): [0029] R is joined from the methylene moiety formed by reduction of the carboxylic acid of one of the following polyunsaturated fatty acids (PUFAs): cis,cis,cis-7,10,13-hexadecatrienoic acid (HTA), cis,cis,cis-9,12,15-octadecatrienoic acid (ALA), cis,cis,cis,cis-6,9,12,15-octadecatetraenoic acid (SDA), cis,cis,cis-11,14,17-eicosatrienoic acid (ETE), cis,cis,cis,cis-8,11,14,17-eicosatetraenoic acid (ETA); cis,cis,cis,cis,cis-5,8,11,14,17-eicosapentanenoic acid (EPA), cis,cis,cis,cis,cis-6,9,12,15,18-heneicosapentaenoic acid (HPA), cis,cis,cis,cis,cis-7,10,13,16,19-docosapentaenoic acid (DPA), cis,cis,cis,cis,cis,cis-4,7,10,13,16,19-docosahexaenoic acid (DHA), cis,cis,cis,cis,cis-9,12,15,18,21-tetracosapentaeonic acid (TPA) or cis,cis,cis,cis,cis,cis-6,9,12,15,18,21-tetracosahexaeonic acid (THA).

[0030] Because the omega-3 acids are already known to be mild PPARγ agonists, the present invention utilizes the formation of compounds by modifying the carboxylic acid of the PUFA and covalently joining an isobutyrate functionality and tests if these compounds have “souped-up” PPARγ activity, and/or other unique biological properties. Such compounds of Formula (I) can be used alone as a pharmaceutically-acceptable formulation, such as a tablet, hard or soft gelatin capsule, or other formulations, or in combination with metformin (Formula II) in treating metabolic syndrome, and also assuring safe cardiovascular health.

Alzheimer' Disease (AD)

[0031] The prevalence and incidence of Alzheimer's disease, and its devastating effects on the lives of patients and care giver families are well known. The health care costs to society are onerous, and will continue to grow with the aging population. Enormous strides have been made in understanding the pathology of the disease which leads to the build-up of amyloid plaques in the brain, which are aggregates of amyloid beta (Aβ) peptides. Fundamental advances have been made in discovering inhibitors of the extra-cellular and intra-cellular neuronal biochemical enzymes such as β-secretage (BACE1) or γ-secretase (GS) to stop the amyloid or intraneuronal τ-tangles build-up; and even reverse these processes through treatment with specific monoclonal antibodies. However, in spite of massive scientific research and investments in reversing the cognitive decline of AD, these have yielded scant benefits. Consensus is emerging that the best approach would be to treat patients before the disease has progressed too far, and even before disease symptoms become apparent. Multi-targeted Alzheimer's drugs, for example dual BACE/acetylcholine esterase inhibition or GSM/PPARγ active agents would offer additional benefits (Harrie J. M. Gisjen, et al., “Secretase Inhibitors and Modulators as a Disease-Modifying Approach Against Alzheimer's Disease”, Annual Reports in Medicinal Chemistry, 2012, 47, 55-69).

[0032] The presence of omega-3 fatty acids, especially DHA in the brain is ubiquitous. Clinical studies in 4 year old children support the beneficial effects of docohexaenoic acid (DHA) on cognitive function (NCT 00351624; 2006-2008; sponsored by Martek BioSciences Corporation). It would be an interesting study to follow such treated children over decades regarding the incidence of onset of symptoms of Alzheimer's disease relative to the untreated group. In the meantime, it is worth exploring in a prospective study, if the DHA fibrate, a PPARγ agonist, either alone or in combination with a gamma secretase modulator (GSM) or other prescribed clinical agents, would slow down the decline of cognitive function in early stage AD patients.

[0033] This invention will be further clarified by a consideration of the following examples of synthesis of compounds of Formula (I) which are intended to be purely exemplary of the present invention.

[0034] The procedures are based on reported literature on the synthesis of fibrates (Om P Goel, U.S. Pat. No. 7,345,190; U.S. Pat. No. 7,770,071).

Scheme 1: Synthesis Overview

[0035] The compounds of the invention were synthesized according to Scheme 1 below in good to excellent yields. The structures were all confirmed by NMR spectra, and HRMS or elemental analysis. Purities were determined by HPLC. All fibrate ethyl esters and fibrate acids are a light yellow or colorless oil. The metformin salts are light yellow to off-white waxes, soluble in chloroform, methylene chloride, DMSO, and partially soluble in diethyl ether.

##STR00018##

Example 1: EPA Alcohol, 1

[0036] ##STR00019##

[0037] An oven-dried 100 mL round bottomed flask was charged with lithium aluminum hydride (0.76 g, 20 mmol) in anhydrous THF (15 mL). The flask was then cooled to 0° C. with an ice-water bath. To this suspension was added drop-wise a solution of EPA ethyl ester (3.30 g, 10 mmol) in THF (10 mL) via syringe under argon. When the addition was complete, the mixture was stirred for 3 hr at 0° C. The reaction was monitored by TLC. After the reaction was complete, it was quenched at 0° C. by slow drop-wise addition of saturated aqueous solution of sodium sulfate (4 mL). The mixture was stirred for 0.5 hr at RT and then filtered through a Büchner funnel. The residue was rinsed with THF. The filtrate and washings were combined and concentrated under reduced pressure to obtain 2.88 g of EPA alcohol as a yellow oil, yield: 100%, and is characterized by the following spectra:

[0038] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.50-5.20 (m, 10H), 3.65 (t, J=6.5 Hz, 2H), 2.92-2.75 (m, 8H), 2.18-2.02 (m, 4H), 1.64-1.54 (m, 2H), 1.64-1.38 (m, 2H), 0.98 (t, J=7.7 Hz, 3H).

[0039] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 131.8, 129.7, 128.4, 128.2, 128.0, 127.93, 127.86, 127.81, 127.68, 126.8, 62.8, 32.3, 26.9, 25.7, 25.6, 25.5, 20.5, 14.3.

Example 2: EPA Bromide, 2

[0040] ##STR00020##

[0041] To a solution of EPA alcohol (2.88 g, 10 mmol, prepared as in Example 1) and carbon tetrabromide (3.65 g, 11 mmol) in anhydrous methylene chloride (20 mL) was added triphenylphosphine (2.89 g, 11 mmol) in 4 portions with an interval of 15 min. in between each portion at 0° C. The resulting reaction mixture was stirred at 0° C. The reaction was monitored by TLC. After 4 hr the reaction mixture was concentrated under reduced pressure. Hexanes (30 mL) were added and the mixture was cooled and filtered to remove triphenylphosphine oxide. The filtrate and washings were concentrated under reduced pressure to give crude product as a yellow oil. Purified by silica gel column chromatography (1% ethyl acetate/heptane) provided EPA bromide 2 as colorless oil (3.27 g), yield: 93% and is characterized by the following spectra:

[0042] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.45-5.22 (m, 10H), 3.41 (t, J=6.8 Hz, 2H), 2.90-2.70 (m, 8H), 2.15-2.00 (m, 4H), 1.95-1.81 (m, 2H), 1.58-1.42 (m, 2H), 0.98 (t, J=7.7 Hz, 3H).

[0043] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 131.8, 129.2, 128.4, 128.2, 128.0, 127.9, 127.7, 126.8, 33.6, 32.3, 28.0, 26.3, 25.6, 25.5, 20.5, 14.3.

Example 3: EPA fibrate ethyl ester, 3 (JIVA-0015)

[0044] ##STR00021##

[0045] Lithium diisopropylamide solution in THF/Heptane/ethylbenzene (11.0 mL, 2M, 22 mmol) was added drop-wise to a solution of ethyl isobutyrate (2.55 g, 21.9 mmol) in dry THF (15 mL) at −78° C. The resulting light yellow solution was stirred at −78° C. for 1 hr, a solution of EPA bromide (2.57 g, 7.31 mmol, prepared as in Example 2) in anhydrous THF (5 mL) was added drop-wise. Then the reaction mixture was stirred and warmed to RT overnight under argon. The reaction mixture was quenched with ice (10 g), and 1N HCl (5 mL), and diluted with ethyl acetate (30 mL). The phases were separated and the aqueous phase was extracted with ethyl acetate (10 mL×2). The organic layers were combined, washed with water (10 mL), brine (10 mL), dried over MgSO.sub.4, and concentrated under reduced pressure. Purification by silica gel column chromatography (2% EtOAc/heptane) provided the desired EPA fibrate ethyl ester 3 as colorless oil (1.92 g, 68% yield) and is characterized by the following data and spectra:

[0046] Chemical Formula: C.sub.26H.sub.42O.sub.2

[0047] Molecular Weight: 386.61

[0048] Chromatographic purity (HPLC): 98.7% (rt=7.797 min, 75-100% MeOH/H.sub.2O over 15 min., Luna C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0049] HRMS (DART-ESI-MS): Calculated for C.sub.26H.sub.46NO.sub.2 (M+NH.sub.4).sup.+: 404.3523; found: 404.3534.

[0050] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.45-5.22 (m, 10H), 4.11 (q, J=7.0 Hz, 2H), 2.90-2.75 (m, 8H), 2.13-2.00 (m, 4H), 1.55-1.46 (m, 2H), 1.38-1.18 (m, 4H), 1.24 (t, J=7.1 Hz, 3H), 1.15 (s, 6H), 0.98 (t, J=7.7 Hz, 3H).

[0051] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 177.7, 131.8, 129.9, 128.34, 128.26, 127.98, 127.95, 127.74, 127.68, 127.55, 126.8, 60.1, 42.1, 40.5, 30.1, 27.1, 25.6, 25.5, 25.1, 24.6, 20.5, 14.2.

Example 4: EPA Fibrate or EPA-Fibric Acid, 4 (JIVA-0018)

[0052] ##STR00022##

[0053] A solution of EPA fibrate ester 3 (1.48 g, 3.83 mmol, prepared as in Example 3) and potassium hydroxide (85%, 0.65 g, 9.85 mmol) in ethanol (10 mL) and water (4 mL) was heated to reflux for 20 hr under argon. The ethanol was evaporated under reduced pressure and the remaining mixture was diluted with water (15 mL). After acidification with aqueous 1N HCl to pH=3, the suspension was extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (10 mL), dried over MgSO.sub.4, and concentrated to give a yellow oil. Purification by silica gel flash chromatography (5% ethyl acetate/heptane) provided desired EPA fibrate 4 (1.30 g, 94% yield) as light yellow oil and is characterized by the following data and spectra:

[0054] Chemical Formula: C24H.sub.38O.sub.2

[0055] Molecular Weight: 358.56

[0056] Chromatographic purity (HPLC): 99.6% (rt=13.067 min, 83-100% MeOH/H.sub.2O over 15 min., Luna C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0057] Elemental analysis: Calculated for C.sub.24H.sub.38O.sub.2: C, 80.39; H, 10.68; found: C, 80.66; H, 10.81.

[0058] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.42-5.20 (m, 10H), 2.90-2.72 (m, 8H), 2.13-2.00 (m, 4H), 1.58-1.50 (m, 2H), 1.41-1.22 (m, 4H), 1.19 (s, 6H), 0.97 (t, J=7.5 Hz, 3H).

[0059] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 184.1, 131.8, 129.9, 128.33, 128.25, 127.97, 127.96, 127.76, 127.69, 127.60, 126.8, 42.0, 40.3, 30.0, 27.0, 25.6, 25.5, 24.9, 24.5, 20.5, 14.2.

Example 5: EPA Fibrate Metformin Salt, 5 (JIVA-0021)

[0060] ##STR00023##

[0061] To a solution of starting EPA fibric acid (0.60 g, 1.67 mmol, prepared as in Example 4) in absolute ethanol (3 mL) was added a solution of metformin (0.24 g, 1.84 mmol) in absolute ethanol (2 mL) at RT under argon. The resulting solution was stirred at RT for 2 hr. The ethanol was removed under reduced pressure under 30° C. The residue was dissolved in ether (3 mL) and cooled at −10° C. overnight. Filtration provided desired salt 5 (0.45 g, 55% yield) as yellow wax and is characterized by the following data and spectra:

[0062] Chemical Formula: C.sub.28H.sub.49N.sub.5O.sub.2

[0063] Molecular Weight: 487.72

[0064] Chromatographic purity (HPLC): 98.9% (rt=14.667 and 19.819 min, 83-100% MeOH/H.sub.2O over 15 min., Alltima C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0065] Elemental analysis: Calculated for C.sub.28H.sub.49N.sub.5O.sub.20.85H.sub.2O: C, 66.86; H, 10.16; N; 13.92;

[0066] found: C, 65.94; H, 10.24; N; 14.87.

[0067] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.80-4.80 (m, 15H), 3.02 (s, 6H), 2.90-2.65 (m, 8H), 2.20-1.95 (m, 4H), 1.50-1.38 (m, 2H), 1.38-1.20 (m, 5H), 1.08 (s, 6H), 0.97 (t, J=7.5 Hz, 3H).

[0068] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 185.5, 160.4, 158.5, 131.9, 130.5, 128.5, 128.4, 128.1, 127.8, 127.3, 126.9, 43.1, 41.7, 37.6, 30.6, 27.4, 26.3, 25.7, 25.1, 20.6, 14.2.

Example 6: ALA Alcohol, 6

[0069] ##STR00024##

[0070] An oven-dried 100 mL round bottomed flask was charged with lithium aluminum hydride (2.04 g, 53.87 mmol) in anhydrous THF (20 mL). The mixture was cooled to 0° C. with an ice-water bath. To this suspension was added drop-wise a solution of ALA (5.00 g, 17.96 mmol) in THF (20 mL) via syringe under argon. When the addition was complete, the mixture was allowed to warm to RT and stirred for 4 hr. The reaction mixture was then quenched at 0° C. by slow drop-wise addition of a saturated aqueous solution of sodium sulfate (10 mL). The mixture was then allowed to stir for 0.5 hr at RT and then filtered through a Büchner funnel. The residue was rinsed with THF. The filtrate and washings were combined and concentrated under reduced pressure to obtain 4.75 g of ALA alcohol as colorless oil (100% yield) and is characterized by the following spectra:

[0071] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.45-5.25 (m, 6H), 3.63 (t, J=6.6 Hz, 2H), 2.81 (t, J=5.7 Hz, 4H), 2.15-2.00 (m, 4H), 1.62-1.45 (m, 3H), 1.42-1.20 (m, 10H), 0.98 (t, J=7.7 Hz, 3H).

[0072] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 131.7, 130.1, 128.1, 127.5, 126.9, 62.9, 32.7, 29.6, 29.5, 29.4, 29.2, 27.2, 25.7, 25.6, 25.5, 20.5, 14.3.

Example 7: ALA Bromide, 7

[0073] ##STR00025##

[0074] To a solution of ALA alcohol (4.75 g, 17.96 mmol) and carbon tetrabromide (6.55 g, 19.76 mmol) in anhydrous methylene chloride (30 mL) was added triphenylphosphine (5.18 g, 19.76 mmol) in 4 portions with an interval of 15 min. in between each portion at 0° C. The resulting reaction mixture was stirred at 0° C. The reaction was monitored by TLC. After 4 hr, the reaction mixture was concentrated under reduced pressure. Hexanes (50 mL) were added and the mixture was cooled and filtered to remove triphenylphosphine oxide. The filtrate and washings were concentrated under reduced pressure to give crude product as yellow oil. Purified by silica gel column chromatography (1% ethyl acetate/heptane) to provide ALA bromide 7 as colorless oil (5.54 g, 94% yield) and is characterized by the following spectra:

[0075] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ5.45-5.25 (m, 6H), 3.40 (t, J=6.9 Hz, 2H), 2.81 (t, J=5.6 Hz, 4H), 2.14-2.00 (m, 4H), 1.95-1.80 (m, 2H), 1.48-1.15 (m, 10H), 0.98 (t, J=7.7 Hz, 3H).

[0076] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 131.7, 130.0, 128.1, 128.0, 127.5, 126.9, 33.9, 32.8, 29.5, 29.3, 29.1, 28.7, 28.1, 27.2, 25.6, 25.5, 20.5, 14.3.

Example 8: ALA Fibrate Ethyl Ester, 8 (JIVA-0016)

[0077] ##STR00026##

[0078] Lithium diisopropylamide solution in THF/Heptane/ethylbenzene (25.3 mL, 2M, 50.6 mmol) was added drop-wise to a solution of ethyl isobutyrate (5.88 g, 50.59 mmol) in dry THF (25 mL) at −78° C. The resulting light yellow solution was stirred at −78° C. for 1 hr, and a solution of ALA bromide (5.52 g, 16.86 mmol) in anhydrous THF (10 mL) was added drop-wise. Then the reaction mixture stirred and warmed to RT overnight under argon. The reaction mixture was quenched with ice (20 g), and 1N HCl (20 mL), and diluted with ethyl acetate (50 mL). The phases were separated and the aqueous phase was extracted with ethyl acetate (20 mL×2). The organic layers were combined, washed with water (20 mL), brine (20 mL), dried over MgSO.sub.4, and concentrated in vacuo. Purification by silica gel column chromatography (2% ethyl acetate/heptane) provided the desired ALA fibrate ethyl ester 8 as light yellow oil (5.90 g, 96% yield) and is characterized by the following data and spectra:

[0079] Chemical Formula: C.sub.24H.sub.42O.sub.2

[0080] Molecular Weight: 362.59

[0081] Chromatographic purity (HPLC): 98.9% (rt=8.619 min, 93-100% MeOH/H.sub.2O over 15 min., Luna C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0082] HRMS (DART-ESI-MS): Calculated for C.sub.24H.sub.46NO.sub.2 (M+NH.sub.4)+: 380.3523; found: 380.3515.

[0083] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.55-5.25 (m, 6H), 4.11 (q, J=7.2 Hz, 2H), 2.85-2.75 (m, 4H), 2.13-2.00 (m, 4H), 1.52-1.45 (m, 2H), 1.40-1.05 (m, 15H), 1.15 (s, 6H), 0.98 (t, J=7.7 Hz, 3H).

[0084] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 177.8, 131.7, 130.2, 128.1, 127.4, 126.9, 60.0, 42.1, 40.7, 30.1, 29.6, 29.4, 29.3, 27.2, 25.6, 25.5, 25.1, 24.9, 20.5, 14.2.

Example 9: ALA Fibrate, 9 (JIVA-0019)

[0085] ##STR00027##

[0086] A solution of starting ALA fibrate ester 8 (3.50 g, 9.65 mmol, prepared as in Example 8) and potassium hydroxide (85%, 1.82 g, 27.5 mmol) in ethanol (30 mL) and water (12 mL) was heated to reflux for 20 hr under argon. The ethanol was evaporated under reduced pressure and the remaining mixture was diluted with water (45 mL). After acidification with aqueous 1N HCl to pH=3, the formed suspension was extracted with ethyl acetate (3×30 mL). The combined organic phase was washed with brine (20 mL), dried over MgSO.sub.4, and concentrated to give yellow oil. Purification by silica gel flash chromatography (5% ethyl acetate/heptane) provided desired ALA fibrate 9 (3.04 g, 94% yield) as light yellow oil and is characterized by the following data and spectra:

[0087] Chemical Formula: C.sub.22H.sub.38O.sub.2

[0088] Molecular Weight: 334.54

[0089] Chromatographic purity (HPLC): 96.0% (rt=11.179 min, 97-100% MeOH/H.sub.2O over 8 min., Alltima C18, 5μ, 4.6×250 mm, 1.0 mL/min, 2 μL injection, 40° C., UV detection, 210 nm)

[0090] Elemental analysis: Calculated for C.sub.22H.sub.38O.sub.2: C, 78.99; H, 11.45; found: C, 79.04; H, 11.30.

[0091] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.45-5.26 (m, 6H), 2.86-2.76 (m, 4H), 2.12-2.00 (m, 4H), 1.58-1.48 (m, 2H), 1.41-1.10 (m, 13H), 1.19 (s, 6H), 0.98 (t, J=7.5 Hz, 3H).

[0092] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 184.4, 131.7, 130.2, 128.1, 128.0, 127.4, 126.9, 42.1, 40.5, 30.1, 29.6, 29.5, 29.4, 29.3, 27.2, 25.6, 25.5, 24.9, 24.8, 20.5, 14.3.

Example 10: ALA Fibrate Metformin Salt, 10 (JIVA-0022)

[0093] ##STR00028##

[0094] To a solution of ALA fibrate (1.90 g, 5.68 mmol, prepared as in Example 9) in absolute ethanol (5 mL) was added a solution of metformin (0.74 g, 5.68 mmol) in absolute ethanol (3 mL) at RT under argon. The resulting solution was stirred at RT for 2 hr. The ethanol was removed under reduced pressure under 30° C. The residue was dissolved in ether (8 mL) and cooled at −10° C. overnight. Filtration provided desired salt 10 (1.61 g, 61% yield) as off-white wax and is characterized by the following data and spectra:

[0095] Chemical Formula: C.sub.26H.sub.49N.sub.5O.sub.2

[0096] Molecular Weight: 463.70

[0097] Chromatographic purity (HPLC): 99.8% (rt=13.067 and 23.339 min, 72-100% MeOH/H.sub.2O over 15 min., Alltima C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0098] Elemental analysis: Calculated for C.sub.26H.sub.49N.sub.5O.sub.20.5H.sub.2O: C, 66.06; H, 10.66; N; 14.82;

[0099] found: C, 66.25; H, 10.49; N; 14.52.

[0100] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.80-4.80 (m, 15H), 3.02 (s, 6H), 2.90-2.65 (m, 8H), 2.20-1.95 (m, 4H), 1.50-1.38 (m, 2H), 1.38-1.20 (m, 5H), 1.08 (s, 6H), 0.97 (t, J=7.5 Hz, 3H).

[0101] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 185.5, 160.4, 158.5, 131.9, 130.5, 128.5, 128.4, 128.1, 127.8, 127.3, 126.9, 43.1, 41.7, 37.6, 30.6, 27.4, 26.3, 25.7, 25.1, 20.6, 14.2.

Example 11: DHA Alcohol, 11

[0102] ##STR00029##

[0103] An oven-dried 100 mL round bottomed flask was charged with lithium aluminum hydride (1.06 g, 28 mmol) in anhydrous THF (15 mL). The flask was cooled to 0° C. with an ice-water bath. To this suspension was added drop-wise a solution of DHA ethyl ester (5.00 g, 14 mmol) in THF (10 mL) via syringe under argon. When the addition was complete, the mixture was allowed to stir for 3 hr at 0° C. The reaction was monitored by TLC. After the reaction was complete, it was quenched at 0° C. by slow drop-wise addition of a saturated aqueous solution of sodium sulfate (5.5 mL). The mixture was then allowed to stir for 0.5 hr at RT and then filtered through a Büchner funnel. The residue was rinsed with THF. The filtrate and washings were combined and concentrated under reduced pressure to obtain 4.41 g of ALA alcohol as light yellow oil (˜100% yield) and is characterized by the following spectra:

[0104] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ5.50-5.25 (m, 12H), 3.66 (t, J=6.5 Hz, 2H), 2.95-2.72 (m, 10H), 2.22-2.05 (m, 4H), 1.70-1.55 (m, 2H), 0.98 (t, J=7.7 Hz, 3H).

[0105] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 131.8, 129.2, 128.4, 128.3, 128.1, 128.0, 127.96, 127.89, 127.7, 126.8, 62.4, 32.4, 25.6, 25.5, 23.6, 20.5, 14.3.

Example 12: DHA Bromide, 12

[0106] ##STR00030##

[0107] To a solution of DHA alcohol (4.41 g, 14 mmol, prepared as in Example 11) and carbon tetrabromide (5.12 g, 15.4 mmol) in anhydrous methylene chloride (30 mL) was added triphenylphosphine (4.04 g, 15.4 mmol) in 4 portions with an interval of 15 min. in between each portion at 0° C. The resulting reaction mixture was stirred at 0° C. The reaction was monitored by TLC. After 4 hr, the reaction mixture was concentrated under reduced pressure. Hexanes (40 mL) were added, and the mixture was cooled and filtered to remove triphenylphosphine oxide. The filtrate and washings were concentrated under reduced pressure to give crude product as yellow oil. Purified by silica gel column chromatography (1% ethyl acetate/heptane) provide DHA bromide 12 as colorless oil (4.77 g, 90% yield) and is characterized by the following spectra:

[0108] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): 65.48-5.20 (m, 12H), 3.41 (t, J=6.6 Hz, 2H), 2.92-2.70 (m, 10H), 2.30-2.15 (m, 2H), 2.15-2.00 (m, 2H), 2.00-1.85 (m, 2H), 0.97 (t, J=7.5 Hz, 3H).

[0109] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 131.8, 129.3, 128.4, 128.1, 128.0, 127.92, 127.87, 127.75, 127.66, 126.8, 33.2, 32.4, 25.6, 25.5, 20.5, 14.3.

Example 13: DHA Fibrate Ethyl Ester, 13 (JIVA-0017)

[0110] ##STR00031##

[0111] Lithium diisopropylamide solution in THF/Heptane/ethylbenzene (4.0 mL, 2M, 8.0 mmol) was added drop-wise to a solution of ethyl isobutyrate (0.92 g, 7.95 mmol) in dry THF (6 mL) at −78° C. The resulting light yellow solution was stirred at −78° C. for 1 hr. A solution of DHA bromide, (1.00 g, 2.65 mmol) in anhydrous THF (2 mL) was added drop-wise. The reaction mixture stirred and warmed to RT overnight under argon. The reaction mixture was quenched with ice (2 g), and 1N HCl (1 mL), and diluted with ethyl acetate (20 mL). The phases were separated and the aqueous phase was extracted with ethyl acetate (10 mL×2). The organic layers were combined, washed with water (10 mL), brine (10 mL), dried over MgSO.sub.4, and concentrated in vacuo. Purification by silica gel column chromatography (2% ethyl acetate/heptane) provided the desired DHA fibrate ethyl ester 13 as light yellow oil (0.67 g, 61% yield) and is characterized by the following data and spectra:

[0112] Chemical Formula: C.sub.2H.sub.44O.sub.2

[0113] Molecular Weight: 412.65

[0114] Chromatographic purity (HPLC): 94.4% (rt=12.683 min, 93-100% MeOH/H.sub.2O over 15 min., Luna C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0115] HRMS (DART-ESI-MS): Calculated for C.sub.28H.sub.48NO.sub.2(M+NH.sub.4).sup.+: 430.3680; found: 430.3689.

[0116] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.55-5.25 (m, 6H), 4.11 (q, J=7.2 Hz, 2H), 2.85-2.75 (m, 4H), 2.13-2.00 (m, 4H), 1.52-1.45 (m, 2H), 1.40-1.05 (m, 15H), 1.15 (s, 6H), 0.98 (t, J=7.7 Hz, 3H).

[0117] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 177.8, 131.7, 130.2, 128.1, 127.4, 126.9, 60.0, 42.1, 40.7, 30.1, 29.6, 29.4, 29.3, 27.2, 25.6, 25.5, 25.1, 24.9, 20.5, 14.2.

Example 14: DHA Fibrate, 14 (JIVA-0020)

[0118] ##STR00032##

[0119] A solution of starting DHA fibrate ester 13 (1.74 g, 4.22 mmol, prepared as in Example 13) and potassium hydroxide (85%, 0.72 g, 10.82 mmol) in ethanol (10 mL) and water (4 mL) was heated to reflux for 20 hr under argon. The ethanol was evaporated under reduced pressure and the remaining mixture was diluted with water (15 mL). After acidification with aqueous 1N HCl to pH=3, the formed suspension was extracted with ethyl acetate (3×20 mL). The combined organic phase was washed with brine (10 mL), dried over MgSO.sub.4, and concentrated to give a yellow oil. Purification by silica gel flash chromatography (5% ethyl acetate/heptane) provided desired DHA fibrate 14 (1.29 g, 80% yield) as a light yellow oil and is characterized by the following data and spectra:

[0120] Chemical Formula: C.sub.26H.sub.40O.sub.2

[0121] Molecular Weight: 384.60

[0122] Chromatographic purity (HPLC): 90.7% (rt=12.096 min, 93-100% MeOH/H.sub.2O over 15 min., Alltima C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0123] Elemental analysis: Calculated for C.sub.22H.sub.38O.sub.2: C, 81.2; H, 10.48; found: C, 80.23; H, 10.38.

[0124] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.45-5.26 (m, 6H), 2.86-2.76 (m, 4H), 2.12-2.00 (m, 4H), 1.58-1.48 (m, 2H), 1.41-1.10 (m, 13H), 1.19 (s, 6H), 0.98 (t, J=7.5 Hz, 3H).

[0125] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 184.4, 131.7, 130.2, 128.1, 128.0, 127.4, 126.9, 42.1, 40.5, 30.1, 29.6, 29.5, 29.4, 29.3, 27.2, 25.6, 25.5, 24.9, 24.8, 20.5, 14.3.

##STR00033##

Example 15: DHA Fibrate Metformin Salt, 15 (JIVA-0023)

[0126] To a solution of starting DHA fibrate (0.52 g, 1.35 mmol, prepared as in Example 14) in absolute ethanol (2 mL) was added a solution of metformin (0.18 g, 1.35 mmol) in absolute ethanol (2 mL) at RT under argon. The resulting solution was stirred at RT for 2 hr. The ethanol was removed under reduced pressure under 30° C. The residue was dissolved in ether (2 mL) and cooled at −10° C. overnight. Filtration provided desired salt 15 (0.34 g, 49% yield) as light yellow wax and is characterized by the following data and spectra:

[0127] Chemical Formula: C.sub.30H.sub.51N.sub.5O.sub.2

[0128] Molecular Weight: 513.76

[0129] Chromatographic purity (HPLC): 94.8% (rt=14.400 and 22.699 min, 72-100% MeOH/H.sub.2O over 15 min., Alltima C18, 5μ, 4.6×250 mm, 1.0 mL/min, 5 μL injection, 40° C., UV detection, 210 nm)

[0130] Elemental analysis: Calculated for C.sub.30H.sub.51N.sub.5O.sub.20.8H.sub.2O: C, 68.22; H, 10.04; N; 13.26;

[0131] found: C, 68.36; H, 10.03; N; 13.13.

[0132] .sup.1H NMR (300 MHz, CDCl.sub.3/TMS): δ 5.80-4.80 (m, 15H), 3.02 (s, 6H), 2.90-2.65 (m, 8H), 2.20-1.95 (m, 4H), 1.50-1.38 (m, 2H), 1.38-1.20 (m, 5H), 1.08 (s, 6H), 0.97 (t, J=7.5 Hz, 3H).

[0133] .sup.13C NMR (75 MHz, CDCl.sub.3/TMS): δ 185.5, 160.4, 158.5, 131.9, 130.5, 128.5, 128.4, 128.1, 127.8, 127.3, 126.9, 43.1, 41.7, 37.6, 30.6, 27.4, 26.3, 25.7, 25.1, 20.6, 14.2.

Utility: In Vitro Biology

[0134] The compounds of the present invention are expected to have beneficial effects on metabolic health by activation of the transcription factor PPARα. Although the molecular details are not fully understood, activation of PPARα transcriptional activity increases fatty acid oxidation in multiple tissues, and that this results in a reduction in circulating fatty acids and triglycerides. The activity of JIVA-0018, JIVA-0019 and JIVA-0020 was determined in a PPARα-dependent transcription assay. This assay was carried out in live cells treated with these above compounds, and provides a direct measurement of a compound's ability to activate PPARα. This PPARα activity assay is a standard nuclear receptor ligand activity assay that utilizes the ligand-binding domain of the PPAR receptor fused to a heterologous GAL4 DNA binding domain. The transcriptional read-out is from a GAL4-regulated luciferase reporter. In this assay, compounds that activate the receptor cause an increase in luciferase activity measured in a luminometer. The activity of the invention compounds was compared to two well-characterized PPARα ligands: bezafibrate and GW7647.

[0135] As shown in FIG. 1, while neither the EPA-fibrate, JIVA-0018 nor the DHA-fibrate, JIVA-0020 stimulated PPARα activity, the ALA-fibrate, JIVA-0019, strongly activated PPARα. From these data the potency (EC.sub.50) for JIVA-0020 activation of PPARα can be estimated to be somewhat greater than 20 micromolar. This is more potent than benzafibrate, which has an EC.sub.50 on PPARα of 100 micromolar in this assay, although it is less potent than GW7647, which has an EC.sub.50 of 11 nanomolar.

[0136] None of the JIVA compounds were active on the related PPAR receptor PPARδ (FIG. 2), and showed only a weak activity on PPARγ (FIG. 3). Thus, it is evident from these data that the ALA-fibrate, JIVA-0019, is a potent and selective activator of PPARα.

[0137] Compounds of Formula (I) are preferably used as a pharmaceutically-acceptable formulation such as pharmaceutically-acceptable adjuvants, binders, desiccants, diluents and excipients that are well known for such purpose. Such formulations are in the form of a solution for injection, ampoule, hard or soft gelatin capsule or tablet, or sustained release formulations. These formulations are used to treat persons for Type2 diabetes, obesity, hypertriglyceridemia, cardiovascular diseases, metabolic syndrome, cancer, Alzheimer's disease: and their use for modulating activity of peroxisome proliferator-activated receptors (PPARs).

[0138] When the person's the triglycerides levels are in a range of from >100 mg/dl, to >500 mg/dl, then the person needs treatment. An effective amount of the active ingredient in the formulations is from about 0.05 to about 5 g/day administered as 1-4 doses/day.

[0139] Although the invention has been described with reference to its preferred embodiments, those of ordinary skill in the art may, upon reading and understanding this disclosure, appreciate changes and modifications which may be made which do not depart from the scope and spirit of the invention as described above or claimed hereafter.