WO2016092479A1

WO2016092479A1 - A pharmaceutical formulation comprising resveratrol, benfotiamine and pyridoxamine, a process of making it and a method of treatment using it

Info

Publication number: WO2016092479A1
Application number: PCT/IB2015/059459
Authority: WO
Inventors: Subramaniam Vanangamudi Sulur; Murali Srinivasan; Madhavan Srinivasan; Sivaraman GOPALAKRISHNAN
Original assignee: Subramaniam Vanangamudi Sulur
Priority date: 2014-12-10
Filing date: 2015-12-09
Publication date: 2016-06-16

Abstract

The invention relates to a stable oral liquid formulation developed for treating negative effects of Advanced Glycation end Products (AGEs) in a clear oral liquid formulation containing Resveratrol, Benfotiamine, Pyridoxamine and one or more pharmaceutically active agents in a suitable vehicle and an effective amount of solubilizer. The solubilizer may include Povidone, Polyoxyl 40 hydrogenated castor oil but not limited to these wherein the solubilizer enhances solubility of Resveratrol in water and stabilizes the Benfotiamine and Pyridoxamine in the formulation along with Resveratrol. The process of making the formulation is unique. It may be used for treatment and prevention of conditions and diseases including many degenerative diseases, such as diabetes, atherosclerosis, chronic renal failure caused due to AGEs and formation of Advanced Lipoxidation End Products (ALEs).

Description

A PHARMACEUTICAL FORMULATION COMPRISING RESVERATROL, BENFOTIAMINE AND PYRIDOXAMINE, A PROCESS OF MAKING IT

AND A METHOD OF TREATMENT USING IT

Field of the Invention

This invention relates to a stable oral liquid formulation developed for treating negative effects of Advanced Glycation end Products (AGEs) in a clear oral liquid formulation containing Resveratrol, Benfotiamine, Pyridoxamine and one or more pharmaceutically active agents in a suitable vehicle and an effective amount of solubilizer. The solubilizer may include Povidone, Polyoxyl 40 hydrogenated castor oil but not limited to these wherein the solubilizer enhances solubility of Resveratrol in water and stabilizes the Benfotiamine and Pyridoxamine in the formulation along with Resveratrol. The process of making the formulation is unique. It may be used for treatment and prevention of conditions and diseases including many degenerative diseases, such as diabetes, atherosclerosis, chronic renal failure caused due to AGEs and formation of Advanced Lipoxidation End Products (ALEs).

Background of the Invention

In human nutrition and biology, the AGEs are substances that can be a factor in the development or worsening of many degenerative diseases, such as diabetes, atherosclerosis, chronic renal failure, and Alzheimer's disease. AGEs are formed inside the human body. They stem from glycation reaction, which refers to the addition of a carbohydrate to a protein without the involvement of an enzyme. Glucose can bind with proteins in a process called glycation, making cells stiffen, less pliable and more subject to damage leading to premature aging.

The animal and human evidence is that significant amounts of AGEs are absorbed, and that the AGEs contribute to the body's burden of AGE, and are associated with diseases such as atherosclerosis and kidney disease.

AGEs are the subject of ongoing research. Compounds that have been found to prevent negative effects and inhibit AGE formation include Benfotiamine, Pyridoxamine and Resveratrol. Studies have proved that Benfotiamine and Pyridoxamine help in inhibiting the formation of AGEs and Resveratrol can prevent the negative effects of the AGEs. These compounds have many proven therapeutic benefits other than age related conditions and are used globally. Piperine is used in the formulation which improves the Bioavailability of Resveratrol multifold. This inclusion also helps to improve the bioavailability of Benfotiamine and Pyridoxamine to a great extent. There are several patents filed globally highlighting the synergistic effect of Resveratrol by the addition of Piperine and which serves as an embodiment to the present invention of combining all the four compounds mentioned above as a single product. This combination also helps in reducing the Gentamycin-induced nephrotoxicity.

The manufacturing process deployed for this oral liquid formulation with the above three active ingredients along with one or more pharmaceutically active agents is unique which has ensured to avoid the interaction between them thus improving the therapeutic response due to the synergistic effect when these three compounds along with one or more pharmaceutically active agents are used together.

List Of Figures:

Figure 1 - formation of micro emulsion

Figure 2 - Flowchart of the process of making the formulation

Objects and advantages of the invention

It is the objective of the present invention to provide a method and formulation for reducing and preventing the negative effects of AGEs.

It is another and more specific objective of the invention to provide a stable oral liquid formulation and method for an AGEs-preventive regiment and/or therapy based on oral intake of an active agent or derivative thereof, in association with an orally acceptable vehicle and other inactive ingredients.

Some of the key features or advantages of the formulation of the invention are that the formulation:

- is sugar-free and hence it can be used extensively by diabetic patients, offers improved bioavailability of Resveratrol by formation of micro emulsion (see Figure 1);

- offers improved bioavailability by retarding the metabolism of the actives

- is an aqueous formulation and not a non-aqueous formulation which conventionally is formulated with alcohol Summary of invention

These objects are accomplished by the present invention, which provides a method for inhibiting the formation of AGEs through an oral administration of an effective amount of a combination of Benfotiamine, Pyridoxamine and Resveratrol along with one or more pharmaceutically active agents in a stable oral liquid formulation with acceptable inactive ingredients. The formulation inhibits the formation of AGEs and the damage caused by chronological aging, damage caused by photo-aging and other degenerative diseases related to the increase in AGEs. In the preferred embodiment of the invention, the combination of Benfotiamine, Pyridoxamine, and Resveratrol along with one or more pharmaceutically active agents is administered as a stable oral liquid formulation containing acceptable vehicle and other inactive ingredients to facilitate oral administration. In some cases, this combination also provides additional therapeutic effects by improving the absorption of the active ingredients. In the preferred embodiment of the invention, the combination of Benfotiamine, Pyridoxamine, and Resveratrol along with one or more pharmaceutically active agents is administered as a stable oral liquid formulation containing acceptable vehicle and other inactive ingredients to facilitate oral administration. This combination also provides highly bio-available absorption of the active ingredients.

Accordingly, the invention discloses an aqueous formulation in which Resveratrol is rendered water-soluble, surprisingly in the presence of Benfotiamine, Pyridoxamine The formulation contains Resveratrol, water, and an effective amount of a solubilizer containing Polyoxyl 40 hydrogenated castor oil and not limited to this. The solubilizer acts to render the Resveratrol water-soluble. The formulation also contains artifical sweeteners, colouring agents and flavouring agents.

The artificial sweeteners are selected from a group of sweeteners comprising neotame, mannitol, acesulfame potassium, aspartame, fructose, maltitol, xylitol, saccharin, sodium cyclamate, sucralose and sucrose, in an amount between 0.05% to 4% w/v.

Colouring agents may also be incorporated in the pharmaceutical formulation to provide an appealing color to the formulation. The colouring agents should be selected to avoid chemical incompatibilities with other ingredients in the formulation. Suitable colouring agents are well known to those skilled in the art. An example of suitable colouring agent is caramel. Colouring agents are added in an amount between 0.01% to 2% Examples of suitable flavoring agents include, but are not limited to, natural and artificial flavors such as red grape flavour or tonovin or mints (i.e., peppermint, etc.), menthol, chocolate, artificial chocolate, bubblegum, both artificial and natural fruit flavors (i.e., cherry, grape, orange, strawberry, etc.) and combinations of two or more thereof. Flavoring agents are generally provided as a minor component of the formulation in amounts effective to provide palatable flavor to the formulations. Typically, flavoring agents are present in amounts in the range of about 0 grams to about 5 grams per 100 ml of the formulation.

Preferably, the Polyoxyl 40 hydrogenated castor oil makes up between 1% and 20% of the formulation by weight; more preferably, the Polyoxyl 40 hydrogenated castor oil makes up about 5% of the formulation by weight. In the present invention, the use of Polyoxyl 40 hydrogenated castor oil as a solubilizer produces a surprisingly total solubility of Resveratrol in water. In certain preferred embodiments, the concentration of Resveratrol solubilized in the formulation is between 20 mg/ml and 30 mg/ml.

In the present invention, the stability of Benfotiamine and Pyridoxamine in the aqueous solution of Resveratrol is rendered by using an effective amount of stabilizer containing (but not limited to) Povidone. Preferably, Povidone makes up between 1% and 10% of the formulation by weight; more preferably, the Povidone makes up to about 5% of the formulation by weight.

In the present invention, the use of Povidone as a stabilizer produces a surprisingly high stability of Resveratrol, Benfotiamine and Pyridoxamine in water. In certain preferred embodiments, the concentration of Benfotiamine stabilized in the formulation is between 20 mg/ml and 30 mg/ml and the concentration of Pyridoxamine stabilized in the formulation is between 20 mg/ml and 30 mg/ml.

Accelerated stability studies conducted on the formulations prepared in accordance with the invention disclosure for assessing its shelf life period of 24 months. It suggests that without the presence of Povidone the clarity of the oral formulation is lost and the drug will be precipitated out immediately.

In the present invention, the synergistic action is achieved due to the combination of Benfotiamine and Pyridoxamine along with Resveratrol and one or more pharmaceutically active agents.

In the present invention, the bioavailability of Resveratrol is increased by the inclusion of piperine.

Preferably, the piperine makes up between 0.001% and 0.100% of the formulation by weight; more preferably, the piperine makes up about 0.05% of the formulation by weight. In the present invention, the use of piperine as a bioavailability enhancer produces a surprisingly higher bioavailability of Resveratrol, Benfotiamine and Pyridoxamine.

The formulation as described above may be used for enhanced prevention and/or therapeutic treatment of damage caused by chronological aging, damage caused by photoaging and other degenerative diseases related to the increase in AGEs.

Detailed Description of Invention

The abbreviations used herein have their conventional meaning within the chemical and biological arts, The term "pharmaceutically acceptable salts" is meant to include salts of the active compounds which are prepared with nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein.

When formulations of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent.

Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthaiic, benzenesulfonic, p- tolylsulfonic, citric, tartaric, methanesulfonic, and the like.

Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like. Certain specific formulations of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts. The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

In addition to salt forms, the present invention provides compounds, which are in a prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the formulations of the present invention.

"Resveratrol," as used herein, includes Resveratroi derived from natural sources such as grape skins, wine, or other botanical sources such as P. cuspidatum or C. quinquangulata, or produced synthetically as 98% trans-Resveratrol, available commercially from Sigma Chemical Co., St. Louis, Mo. Botanical extracts with higher concentrations may be produced by fractionation and further column chromatography until an extract may contain up to a 99% concentration of Resveratrol as a mixture of both isomers. "Trans-Resveratrol," as used herein, may also be commonly referred to as trans-3,5,4'-trihydroxystilbene, 3,4',5- stilbenetriol, (E)- 5-(pOhydroxystyryl) resorcinol), and CAS Number 501-36-0. "Benfotiamine," as used herein, includes Benfotiaminemay also be commonly referred to as S-benzoylthiamine O-monophosphate, and CAS Number 22457-89- 2 is a synthetic S-acyl derivative of thiamine.

"Pyridoxamine," as used herein, includes Pyridoxamine may also be commonly referred to as 4-(aminomethyl)-5-(hydroxymethyl)-2-methylpyridin-3-ol, and

CAS Number 85-87-0 is one form of Vitamin Be. Chemically it is based on a pyridine ring structure, with hydroxy!, methyl, aminomethyl, and hydroxymethyl substituents. It differs from pyridoxine by the substituent at the 4-position. The phenol position 3 and aminomethyl group at position 4 of its ring endow Pyridoxamine.

The term "combination therapy" or "adjunct therapy" means that the patient in need of the drug is treated or given another drug for the disease in conjunction with the formulations of the present invention. This combination therapy can be sequential therapy where the patient is treated first with one drag and then the other or the two drugs are given simultaneously. The present invention also discloses a combination therapy or adjunct therapy that uses a stable water-soluble sugar-free formulation containing Resveratrol, Benfotiamine, Pyridoxamine. The term "treating" refers to any indicia of success in the treatment or amelioration of an injury, pathology or condition, including any objective or subjective parameter such as abatement; remission, diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating, improving a patient's physical or mental well-being. The treatment or amelioration of symptoms can be based on objective or subjective parameters, including the results of a physical examination, neuropsychiatric exams, and/or a psychiatric evaluation. For example, the methods of the invention successfully treat a patient's delirium by decreasing the incidence of disturbances in consciousness or cognition.

The term "Patient" refers to a mammalian subject, including human.

The term "clear aqueous formulation" in reference to a solution containing Resveratrol means a water containing solution (e.g. a beverage) that is free of visible particles of un-dissolved Resveratrol. In some embodiments, the clear aqueous formulation not a dispersion and not a suspension, and remains clear upon sitting undisturbed for 1 hour or more. Where the clear aqueous formulation is a beverage, the clear aqueous formulation may not need to be shaken prior to consuming.

The term "cancer" refers to all types of cancer, neoplasm, or malignant tumors found in mammals, including leukemia, carcinomas and sarcomas. Exemplary cancers include cancer of the brain, breast, cervix, colon, head & neck, liver, kidney, lung, non-small cell lung, melanoma, mesothelioma, ovary, sarcoma, stomach, uterus and Medulloblastoma. Additional examples include, Hodgkin's Disease, Non-Hodgkin's Lymphoma, multiple myeloma, neuroblastoma, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, cancer, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, lymphomas, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms of the endocrine and exocrine pancreas, and prostate cancer.

The term "diabetic retinopathy" refers to an ocular pathology associated with diabetes. Diabetes can cause damage to the blood vessels that nourish the retina, and this can cause the vessels to leak or break, stimulating the growth of abnormal new blood vessels. Diabetic retinopathy is one of the leading causes of blindness in diabetics, and affects more than 4 million adults in America alone. The term, 'limiting the progression of renal disease" means to reduce or prevent decreases in renal function in those patients receiving treatment relative to diabetic patients not receiving the treatment. Such treatment thus reduces the need for kidney dialysis or transplantation in diabetic patients. The term "wet age-related macular degeneration (AMD)" refers to an eye condition or disease in which damaging new blood vessel growth and leakage occurs in the retina, and if left untreated can lead to vision loss. AMD is the leading cause of age related blindness. The term, "limiting the progression" of diabetic complications means slowing or stopping the progression of diabetic complications in those patients receiving treatment relative to diabetic patients not receiving the treatment. Thus, the methods of the invention can be used, for example, to slow or stop the progression of nephropathy, neuropathy, retinopathy, and/or symptoms due to impaired microvascular (e.g. erectile dysfunction, angina, claudication) or macrovasular (MI, CVA, amputation, etc.) complications of diabetes in diabetic patients receiving treatment relative to diabetic patients not receiving such treatment.

The term, "nephropathy" refers to kidney disease, inflammation, or damage, "neuropathy" refers to a disease, inflammation, or damage to the nervous system; symptoms include numbness, tingling, pain, or muscle weakness, depending on the neives affected. In a further preferred embodiment, the methods serve to limit one or more symptoms of neuropathy selected from the group consisting of areflexia (reflexes absent), hyporeflexia (weakened reflexes), paresthesia (abnormal sensation, such as burning, pricking, or numbness), peripheral neuropathy (disease, inflammation, or damage to the peripheral nervous system), aggravated peripheral neuropathy, and sensory loss (partial or complete loss of sensory function). As used herein, "retinopathy" refers to a disease, inflammation, or damage to the retina. In the preferred embodiment of the invention, the combination of Benfotiamine, Pyridoxamine, and Resveratrol along with one or more pharmaceutically active agents is administered as a stable oral liquid formulation containing acceptable vehicle and other inactive ingredients to facilitate oral administration. In some cases, this combination also provides additional therapeutic effects by improving the absorption of the active ingredients. In the preferred embodiment of the invention, the combination of Benfotiamine, Pyridoxamine, and Resveratrol along with one or more pharmaceutically active agents is administered as a stable oral liquid formulation containing acceptable vehicle and other inactive ingredients to facilitate oral administration. This combination also provides highly bio-available absorption of the active ingredients.

Accordingly, the invention discloses an aqueous formulation in which Resveratrol is rendered water-soluble, surprisingly in the presence of Benfotiamine, Pyridoxamine The formulation contains Resveratrol, water, and an effective amount of a solubilizer containing Polyoxyl 40 hydrogenated castor oil and not limited to this. The solubilizer acts to render the Resveratrol water-soluble. The formulation also contains artifical sweeteners, colouring agents and flavouring agents. The invention also discloses methods for using such formulations in human diabetic patients, patients with renal disease.

The artificial sweeteners are selected from a group of sweeteners comprising neotame, mannitol, acesulfame potassium, aspartame, fructose, maltitol, xylitol, saccharin, sodium cyclamate, sucralose and sucrose, in an amount between 0.05% to 4%.

Colouring agents may also be incorporated in the pharmaceutical formulation to provide an appealing color to the formulation. The colouring agents should be selected to avoid chemical incompatibilities with other ingredients in the formulation. Suitable colouring agents are well known to those skilled in the art. An example of suitable colouring agent is caramel. Colouring agents are added in an amount between 0.01% to .2%

Examples of suitable flavoring agents include, but are not limited to, natural and artificial flavors such as red grape flavour or tonovin or mints (i.e., peppermint, etc.), menthol, chocolate, artificial chocolate, bubbiegum, both artificial and natural fruit flavors (i.e., cherry, grape, orange, strawberry, etc.) and combinations of two or more thereof Flavoring agents are generally provided as a minor component of the formulation in amounts effective to provide palatable flavor to the formulations. Typically, flavoring agents are present in amounts in the range of about 0 grams to about 5 grams per 100 ml of the formulation. Preferably, the Polyoxyl 40 hydrogenated castor oil makes up between 1% and 20% of the formulation by weight; more preferably, the Polyoxyl 40 hydrogenated castor oil makes up about 5% of the formulation by weight. In the present invention, the use of Polyoxyl 40 hydrogenated castor oil as a solubilizer produces a surprisingly total solubility of Resveratrol in water. In certain preferred embodiments, the concentration of Resveratrol solubilized in the formulation is between 20 mg/ml and 30 mg/ml.

Therapeutic approaches to treating diabetic nephropathy currently follow two strategies: the use of antihypertensive medications to treat hemodynamic factors, and the use of drugs to control blood glucose and the consequences of hyperglycemia (metabolic factors). It has been found that antihypertensive agents can retard the progression of diabetic nephropathy by lowering renal intra- glomerular pressure. Blockade of the renin-angiotensin system is currently the most common approach to achieve this. The angiotensin converting enzyme (ACE) inhibitor, captopril, was first approved for this indication in type 1 diabetes, but it and other ACE inhibitors are routinely also prescribed for nephropathy in type 2 diabetes. Very recently, blockade of the angiotensin 2 (type 1) receptor (ARB) has been demonstrated to have value, with losartan and irbesartan getting FDA approval for the treatment of nephropathy due to type 2 diabetes mellitus. Other modalities include use of diuretics (thiazides), beta blockers and calcium blockers. However, it is recognized that these treatments generally retard but do not prevent the progression of diabetic renal disease beyond their anti-hypertensive actions. The second approach to treatment is to treat metabolic factors associated with elevated glucose (hyperglycemia). Strict glucose control is attempted with insulin, insulin sensitizers, insulin secretalogues, metformin, inhibitors of glucose absorption from the gastrointestinal tract and similar medications. However, perfect glucose control cannot be achieved, and it is recognized that even diabetics maintaining excellent glucose control will still experience damaging fluctuations of their glucose in the blood. Other medications are being developed to halt damage from hyperglycemia, such as protein kinase C inhibitors, superoxide dismutase (SOD) and SOD mimetics, thiamine pyrophosphate and its prodrugs, transketoiase inhibitors, other AGE inhibitors that can mechanistically complement post-Arnadori-inhibitors, glucosaminoglycans, and aldose reductase inhibitors.

As used herein, "diabetic patient" encompasses both Type I and Type 2 diabetic patients and "diabetes" encompasses both Type 1 and Type 2 diabetes.

A newer approach that can be combined with all the metabolic and hemodynamic therapies is to use agents that halt the direct damage that glucose causes to proteins. Pyridoxamine, Benfotiamine, Resveratrol were the most promising of this class of compounds designed as inhibitors of the formation of toxic AGEs that contribute to diabetic complications. Pyridoxamine, Benfotiamine and Resveratrol can be used with these other medications to optimize treatments of general patient populations or with specific patient subpopulations that resist treatment by these other modalities. For example, it is recognized that not all patients tolerate ACE inhibitors or respond to them, This combination with Pyridoxamine, Benfotiamine and Resveratrol may prove to be superior to these therapies. Such co-administration of current therapeutics with Pyridoxamine, Benfotiamine and Resveratrol may also permit administration of lower dose AGEs of these other therapeutics, thus minimizing potential side effects.

Thus, in a further aspect, the present invention provides pharmaceutical formulations comprising (a) Pyridoxamine, or a pharmaceutically acceptable salt thereof: (b) Benfotiamine, or a pharmaceutically acceptable salt thereof: (c) Resveratrol, or a pharmaceutically acceptable salt thereof and (d) one or more compounds that can provide hemodynamic and/or metabolic improvement in a human patient, or pharmaceutically acceptable salts thereof.

In a preferred embodiment, such compounds are selected from the group consisting of angiotensin converting enzyme inhibitors (ACE-I), angiotensin receptor blockers (ARB), beta-blockers, aldose reductase inhibitors, calcium blockers, diuretics, glycosaminoglycans, incretinmimetics, insulin, insulin sensitizers, statins, fibrates, glucose uptake inhibitors, sulfonylureas, superoxide dismutase (SOD) and SOD mimetics, thiamine pyrophosphate and its prodrugs, transketoiase inhibitors, other AGE inhibitors that can mechanistically complement post-Amadori-inhibitors, and protein kinase C inhibitors, The combination of such compounds with Pyridoxamine, Benfotiamine and Resveratrol is demonstrated herein to be effective for limiting the progression of renal disease and diabetic complications in human diabetic patients. In a preferred embodiment of this aspect of the invention, the one or more compounds are selected from the group consisting of angiotensin converting enzyme inhibitors and angiotensin receptor blockers, or pharmaceutically acceptable salts thereof, in a pharmaceutically acceptable carrier.

Non-limiting examples of angiotensin converting enzyme inhibitors for use in the present invention include benazepril, benazeprilat, captopril, delapril, fentiapril, fosinopril, libenzapril, moexipril, pentopril, perindopril, pivopril, quinapril, quinapril at, ramipril, spirapril, spiraprilat, zofenopril, ceronapril, enalapril, indolapril, lisinopril, alacepril, and cilazapril, or pharmaceutically acceptable salts thereof. Non-limiting examples of angiotensin receptor blockers for use in the present invention include losartan, candesartan, irbesartan, olmesartan, valsartan, telmi sartan, eprosartan, and tasosartan.

The progression of renal disease can be measured in various ways, including the following:

(a) Proteinuria (i.e. : increased loss of protein into the urine; often assessed by measurement of albumin levels (i.e. : "albuminuria"));

(b) Impaired glomerular filtration (i.e. : kidney function to clear substances from blood; can be measured, for example, by creatinine (i.e. : "impaired creatinine clearance"), inulin, or urea clearance),

(c) Increased levels of serum creatinine, and

(d) Increased levels of urinary transforming growth factor beta (TGF-β).

Thus, the methods of the invention can be used, for example, to limit the increase in one or more of proteinuria, albuminuria, serum creatinine levels, and urinary TGF-β levels, and/or to limit the impairment of glomemlar filtration and/or creatinine clearance in a diabetic patient being treated with Pyridoxamine, a pharmaceutically acceptable salt thereof, or one of the pharmaceutical formulations of the invention relative to a diabetic patient not receiving such treatment. As will be understood by those of skill in the art, a favorable effect of the methods of the invention on any one or more of these measures of renal disease constitutes limiting the progression of renal disease. In a further embodiment of the methods of the invention, the methods further comprise administering the Pyridoxamine, or a pharmaceutically acceptable salt thereof, Benfotiamine, or a pharmaceutically acceptable salt thereof and Resveratrol, or a pharmaceutically acceptable salt thereof in combination with a further therapeutic to limit the progression of renal disease in a human diabetic patient. Such therapeutics include, but are not limited to, angiotensin converting enzyme inhibitors (ACE- 1), angiotensin receptor blockers (ARB), beta-blockers, aldose reductase inhibitors, calcium blockers, diuretics, glycosaminoglycans, incretinmimetics, insulin, insulin sensitizers, statins, fibrates, glucose uptake inhibitors, sulfonylureas, superoxide dismutase (SOD) and SOD mimetics, thiamine pyrophosphate and its prodrugs, transketolase inhibitors, other AGE inhibitors that can mechanistically complement post-Amadori-inhibitors, and protein kinase C inhibitors. The further therapeutic can be administered together as a single formulation with or separately from the Pyridoxamine, or a pharmaceutically acceptable salt thereof. In a preferred embodiment, the one or more further therapeutics comprise ACE-I and/or ARBs.

In a further preferred embodiment, the human diabetic patient is one that has failed to adequately respond to treatment with ACE-I and/or ARBs. As used herein, "failed to respond adequately" means that one or more measures of the progression of renal disease (proteinuria, albuminuria, serum creatinine levels, impaired glomerular fi ltration, impaired creatinine clearance) continue to increase despite treatment with the ACE-I and/or ARBs.

It is known that formulations containing Pyridoxamine helps human diabetic patients having elevated blood lipid levels, including hyperlipidemia, hypertriglyceridemia, and/or hypercholesterolemia. Such patients tend to have accelerated progression of renal disease relative to other diabetic patients, and the data presented herein demonstrate that treatment of these patients with Pyridoxamine is more effective than treatment with the current standard of care for diabetic kidney disease.

While not being bound by any specific mechanism of action, it is believed that the beneficial effects of the methods of the invention may be due to the inhibitory effect of Pyridoxamine, Benfotiamine and Resveratrol on the formation of AGEs.

The experiments detailed below provide a striking demonstration of the efficacy of Pyridoxamine hydrochloride, Benfotiamine and Resveratrol on limi ting renal disease progression in human diabetic patients compared to patients treated with a placebo. It should be noted that the placebo included the current standard of care for human diabetic patients: Antihypertensive care including the use of ACE-I or ARB treatment and treatment of hyperglycemia and hyperlipidemia, while Pyridoxamine hydrochloride, Benfotiamine and Resveratrol was coadministered with ACE-I and/or ARB. Therefore, the beneficial effect of Pyridoxamine hydrochloride, Benfotiamine and Resveratrol is in addition to any benefit the patient would receive via administration of the current standard of care.

Process Innovation to formulate a stable dear water-soluble sugar-free formulation

It has been discovered that non-ionic surfactants may be used to increase the solubility and/or bioavailability of Resveratrolmetabolites, and salts thereof (e.g. a pharmaceutically acceptable salt) in the water-soluble sugar-free formulations containing Resveratrol, Benfotiamine and Pyridoxamine, a metabolite, or salt thereof. It has been discovered that Resveratrol as an aqueous water-soluble sugar-free formulation along with non-ionic surfactants was stabilized by forming microemulsions using propylene glycol and glycerin under high speed homogenizing to stabilize water-soluble sugar-free formulations containing Resveratrol, Benfotiamine and Pyridoxamine, a metabolite, or salt thereof.

It has been discovered that incorporation of clear aqueous formulation of pipeline under high speed homogenizing into the water-soluble sugar-free formulations containing Resveratrol, Benfotiamine and Pyridoxamine, a metabolite, or salt thereof will improve the bioavailability of Resveratrol, Benfotiamine and Pyridoxamine to achieve the enhanced efficacy. It has been discovered that the solubility of Benfotiamine is improved in presence of alkalinizing agents. The alkalinizing agents is preferably sodium hydroxide but not limited to it.

The stability of the Benfotiamine in the water-soluble sugar-free formulations containing Resveratrol, Benfotiamine and Pyridoxamine, a metabolite, or salt thereof is improved by the presence of (but not limited to) Propylene glycol, Glycerin and non-ionic surfactant like Polyoxyl 40 Hydrogenated Castor Oil.

The stability of the water-soluble sugar-free formulation is further increased by the presence of water-soluble polymer preferable polyvinylpyrollidone made from monomer N-vinylpyrrolidone but not limited to it.

In some embodiments, the water or visible macro-micelles (micelles visible to the naked eye) in water. In other embodiments, the water-soluble sugar-free formulation does not include an alcohol (e.g. the daig is not first dissolved in alcohol and then added to water). A "non-ionic surfactant," as used herein, is a surface active agent that tends to be non-ionized (i.e. uncharged) in neutral solutions (e.g. neutral aqueous solutions). Useful non- ionic surfactants include, for example, non-ionic water-soluble mono- , di-, and tri- glycerides; non-ionic water-soluble mono- and di- fatty acid esters of polyethvelene glycol; non-ionic water-soluble sorbitan fatty acid esters (e.g. sorbitanrnonooleates such as SPAN 80 and TWEEN 20 (polyoxyethylene 20 sorbitanmonooleate)); poiyglycolyzed glycerides, non- ionic water-soluble triblock copolymers (e.g. poly(ethyleneoxide)/poly-(propyleneoxide)/ poly(ethyleneoxide) triblock copolymers such as POLOXAMER 406 (PLURONIC F- 127), and derivatives thereof.

Examples of non-ionic water-soluble mono-, di-, and tri- glycerides include propylene glycol dicarpyiate/dicaprate (e.g. MIGLYOL 840), medium chain mono- and diglycerides (e.g. CAPMUL and IMWITOR 72), medium-chain triglycerides (e.g. caprylic and capric triglycerides such as LAVRAFAC, MIGLYOL 810 or 812, CRODAMOL GTCC- PN, and SOFTISON 378), long chain monoglycerides (e.g. glyceryl nionooieates such as PECEOL, and glyceryl monolinoleates such as MAISINE), Polyoxyl castor oil (e.g. macrogolglycerolricinoleate, macrogolglycerolhydroxystearate, maerogoieetostearyl ether), and derivatives thereof.

Non-ionic water-soluble mono- and di- fatty acid esters of polyethvelene glycol include d-a-tocopherylpolyethyleneglycol 1000 succinate (TPGS), poyethyleneglycol 660 12-hydroxystearate (SOLUTOL HS 15), Polyoxyloleate and stearate (e.g. PEG 400 monostearate and PEG 1750 monostearate), and derivatives thereof. Polyglycoiyzed glycerides include poiyoxyethylated oleic glycerides, polyoxyethylated linoleic glycerides, polyoxyethylatedcaprylic/capric glycerides, and derivatives thereof. Specific examples include LABRAFIL M-1944CS, LABRAFIL - 2125CS, LABRASOL, SOFTIGEN, and GELUCIRE. In some embodiments, the water-soluble sugar-free formulations includes the Resveratrol, Benfotiamine, Pyridoxamine, metabolite or salt thereof, and Polyoxyl castor oil to form a transparent water-soluble sugar-free formulation. A "transparent water-soluble sugar-free formulation," as disclosed herein, refers to a formulation that can be clearly seen through with the naked eye and is optionally colored.

In some embodiments, the transparent water-soluble sugar-free formulations do not contain particles (e.g. particles of undissolved drug) visible to the naked eye. In certain embodiments, light may be transmitted through the transparent water- soluble sugar-free formulations without diffusion or scattering. Thus, in some embodiments, the transparent water-soluble sugar-free formulations are not opaque, cloudy or milky-white. Transparent water-soluble sugar-free formulations disclosed herein do not include milky-white emulsions or suspensions in vegetable oil such as corn oil. It is important to note that transparent water-soluble sugar-free formulations are also typically not formed by first dissolving the drug in alcohol, and then mixed with water, In some embodiments, the water-soluble sugar-free formulation is a non-alcoholic formulation. A "non-alcoholic" formulation, as used herein, is a formulation that does not include (or includes only in trace amounts) methanol, ethanol, propanol or butanol. In other embodiments, the formulation does not include (or includes only in trace amounts) ethanol.

The water-soluble sugar-free formulation of the present invention includes formulations dissolved in water (i.e. aqueous formulations).

In some embodiments, the water-soluble sugarfree formulation forms a transparent water soluble sugar-free formulation when added to water.

In some embodiments, the water-soluble sugar-free formulation consists essentially of Resveratrol (e.g. trans-Resveratrol), Benfotiamine, Pyridoxamine, metabolite or salt thereof, and a non-ionic surfactant, alkalinizing agent, water- soluble polymer and co-solvents to improve the stability. Optionally additional components widely known in the art to be useful in neutraceuticalor drug formulations, such as preservatives, taste enhancers, buffers, water, etc, A water soluble sugar-free formulation that "consists essentially of Resveratrol (e.g. trans- Resveratrol), Benfotiamine and Pyridoxamine, metabolite or salt thereof does not include components that would destroy the novelty and inventiveness of the formulation. Process of making the formulation:

Trans-Resveratrol was dissolved in a mixture of Propylene glycol and Polyoxyl 40 Hydrogenated Castor Oil using high shear homogenizer. To the above solution glycerol and water was added under high speed homogenizing to stabilize the trans-Resveratrol by forming microemulsion. In this microemulsion glycerol and propylene glycol acts as cosolvents. Piperine was dissolved in the water and added to the above solution while homogenizing. Benfotiamine has a poor aqueous solubility but its solubility increase with increase in pH of the aqueous solution. Benfotiamine was dissolved in aqueous alkaline solution of sodium hydroxide with pH between 8 to 9. The above Benfotiamine solution was added to the mixture of Propylene glycol and Polyoxyl 40 Hydrogenated Castor Oil followed by addition of Glycerol. This helps to stabilize the Benfotiamine in the aqueous solution. The above Benfotiamine solution was mixed with the trans-Resveratrol solution slowly under continuous high speed homogenizer. This allows forming a stable molecular dispersion of Benfotiamine and trans-Resveratrol in aqueous solution. The above molecular dispersion of Benfotiamine and trans-Resveratrol was transferred to the aqueous solution of Povidone. The presence of Povidone helps to stabilize the molecular dispersion of Benfotiamine and trans-Resveratrol and reduce the interaction between them in the aqueous solution. The above solution was transferred to sorbitol solution which acts as a base. Pyridoxamine Di-hydrochloride was dissolved in water and transferred to the final formulation.

The resulting formulation may be visually inspected for colloidal particles to determine the degree of solubility of Resveratrol, Benfotiamine and Pyridoxamine along with one or more pharmaceutically active agents.

Referring to Figure 2, the process of making the formulation of the invention comprises the following steps:

a. mixing together purified water and Povidone by stirring at 50 RPM and adding the mixture to a first tank and stirring it to form a homogeneous solution,

b. mixing PGL and Piperine and adding the mixture to a to a second tank and stirring at 50 RPM to form a homogeneous solution,

c. adding in sequence, a. a mixture of Resveratrol and PGL, b. Polyoxyl 40 hydrogenated caster oil, and c. glycerin, to said second tank and stirring it at 50 RPM to form a homogeneous solution,

d. transferring the homogeneous solution of step c and purified water to a third tank and stirring the mixture at 600 RPM to form a homogeneous solution, e. adding a mixture of purified water and sodium hydroxide stored in a fourth tank to a fifth tank and stirring it at 50 RPM to form a homogeneous solution, adding in sequence a. a mixture of benfotiamine and water, b. a mixture of PGL and Polyoxyl 40 hydrogenated caster oil, and c. glycerine to said fifth tank and stirring the contents at 50 RPM to form a homogeneous solution, transferring the mixture of step f along with purified water to said third tank and stirring the contents of said third tank and stirring the mixture at 600 RPM to form a homogeneous solution,

transferring the contents of said third tank to said first tank and stirring the contents of said first tank to form a homogeneous solution,

transferring the contents of said first tank to a sixth tank and stirring the mixture at 900 RPM to form a homogeneous solution,

adding to said sixth tank in sequence, a. sorbitol solution (70%) N- crystalising, b. a mixture of purified water and sodium benzoate, c. a mixture of hot purified water and neotame, d. a mixture of purified water and citric acid monohydrate, and stirring the mixture at 400 RPM to form a

homogeneous solution,

further adding to said sixth tank in sequence a. mixture of purified water and pyridoxamine hydrochloride, b. a mixture of purified water, a first colour and a second colour, c. a mixture of purified water, FAC, manittol, and cyanocobalamin, d. a mixture of a first flavour, PGL, Polyoxyl 40 hydrogenated caster oil, and glycerin, and stirring the mixture at 900 RPM to form a homogeneous solution,

Further adding to said sixth tank tonovin and stirring the contents of said sixth tank at 900 RPM to form a homogeneous solution, m. passing the contents of said sixth tank through a press filter and mix the filtered liquid with a final volume of purified water and stirring the mixture at 1000 RPM to form a homogeneous solution.

An example of the formulation prepared by the process described above provided in the following Table 1 :

IP - Indian Pharmacopoeia

USP - Unites States Pharmacopoeia

IHS - In House Specification

Pharmacological Summary

Methods of treatment using the formulation:

In another aspect, the present invention provides a method of treating cancer, obesity, diabetes, cardiovascular disease, dyslipidaemia, age-related macular degeneration (e.g. vision loss associated with age-related macular degeneration), high cholesterol, or retinopathy (e.g. diabetic retinopathy) in subject in need of such treatment. The method includes administering to the subject an effective amount of the water soluble sugar-free formulations disclosed herein. The term "cancer" is defined in detail above.

In some embodiments, a method of lowering cholesterol in a subject in need of cholesterol lowering therapy is provided. The method includes administering to the subject an effective amount of the water soluble sugar- free formulations disclosed herein. The cholesterol lowering may be total cholesterol lowering or low density lipoprotein (LDL) lowering.

In another aspect, the present invention provides a method of treating a VEGF- mediated disease state in a subject in need of such treatment. The method includes administering to the subject an effective amount of the water soluble sugar- free formulations disclosed herein.

In some embodiments, a method is provided for reducing VEGF-mediated vascular permeability and/or abnormal blood vessel growth in the retina of a subject in need of such treatment. The method includes administering to the subject an effective amount of the water soluble sugar-free formulations disclosed herein. In other embodiments, a method is provided for treating age-related macular degeneration in a subject in need of such treatment. The method includes administering to the subject an effective amount of the water soluble sugar- free formulations disclosed herein.

In still other embodiments, a method is provided for treating diabetic macular edema in a subject in need of such treatment. The method includes administering to the subject an effective amount of the water soluble sugar- free formulations disclosed herein.

Vascular endothelial growth factor (VEGF) is a diffusible protein that is specific to vascular endothelial cells and plays a major role in the regulation of physiological and pathological growth of blood vessels. VEGF promotes the growth of vascular endothelial cells that reside in arteries, veins, and lymphatics, but also has the ability to induce vascular leakage. This permeability enhancing activity is a connecting link between this molecule and other pathological states. For example, VEGF is expressed in the majority of human tumors and plays a critical role in tumor angiogenesis and metastasis. In addition, VEGF is directly involved in the pathological process that leads to the cancer, vision loss associated with age- related macular degeneration (including wet age-related macular degeneration), and retinopathies (such as diabetic retinopathy/diabetic macular edema). Therefore, in some embodiments, a method of reducing the activity of VEGF is provided. The method may be conducted in vitro or in situ for research purposes by contacting VEGF with the water soluble sugar-free formulation of the present invention. Alternatively, the activity of VEGF may be reduced in a subject by administering to the subject an effective amount of the water soluble sugar- free formulation of the present invention.

VEGF inhibition can be measured in- vitro in a suitable cell line such as KOP2.16 endothelial cells, or using other techniques such as the Miles assay.

In another aspect, the present invention provides a method for enhancing the bioavailability of Resveratrol, Benfotiamine and Pyridoxamine, metabolite in a subject in presence of pipeline. The method includes combining Resveratrol, Benfotiamine and Pyridoxamine, metabolite or salt thereof and a non-ionic surfactant to form a surfactant-drug mixture along with polyvinylpyrollidone and co solvents including piperine. The final mixture may be administered to the subject thereby enhancing the bioavailability of the Resveratrol, Benfotiamine and Pyridoxamine, metabolite and salt thereof. The bioavailability is enhanced compared to the bioavailability of Resveratrol, Benfotiamine and Pyridoxaminein the absence of piperine. A subject is an organism that is treated using one of the methods of the present invention. In some embodiment, the subject is a mammalian subject, such as a human or domestic animal. An effective amount of the water soluble sugar-free formulation of the present invention is an amount sufficient to achieve the intended purpose of a method of the present invention, such as treating a particular disease state in a subject (e.g. a human subject). Dosages and Dosage Forms

The amount of Resveratrol, Benfotiamine, Pyridoxamine adequate to treat a disease (e.g. through modulation of VEGF, COX, cell proliferation), is defined as a "therapeutically effective dose". The dosage schedule and amounts effective for this use, i.e., the "dosing regimen," will depend upon a variety of factors, including the stage of the disease or condition, the severity of the disease or condition, the general state of the patient's health, the patient's physical status, age and the like. In calculating the dosage regimen for a patient, the mode of administration also is taken into consideration. The dosage regimen also takes into consideration pharmacokinetics parameters well known in the art, i.e., the rate of absorption, bioavailability, metabolism, clearance, and the like. The state of the art allows the clinician to determine the dosage regimen for each individual patient and disease or condition treated. Single or multiple administrations of Resveratrol, Benfotiamine and Pyridoxamine formulations can be administered depending on the dosage and frequency as required and tolerated by the patient. The formulations should provide a sufficient quantity of active ingredient to effectively treat the disease state.

In some embodiments, the Resveratrol is present in the water-soluble sugar-free formulation at a concentration of at least 0.01%, 0.05%, 0.1%, 0.2%, 0.4%, 0.5%, 0.7%, 0.8%, 0.9%, or 1% by weight, the Benfotiamine is present in the water- soluble sugar-free formulation at a concentration of at least 0.02%, 0.1%, 0.2%, 0.4%, 0.8%, 1%, 1.4%, 1.6%, 1.8%, or 2%> by weight and the Pyridoxamine is present in the water-soluble sugar-free formulation at a concentration of at least 0.01%, 0.05%, 0.1%, 0.2%, 0.4%, 0.5%, 0.7%, 0.8%, 0.9%, or 1% by weight.

In some embodiments, the water soluble sugar-free formulation is in the form of a pharmaceutical formulation. The pharmaceutical formulation may include Resveratrol, Benfotiamine, Pyridoxamine, metabolite, a non-ionic surfactant, water-soluble polymer, co solvents and a pharmaceutically acceptable excipient. After a pharmaceutical formulation including Resveratrol, Benfotiamine, Pyridoxamine of the invention has been formulated in an acceptable carrier, it can be placed in an appropriate container and labeled for treatment of an indicated condition. Unique Advantage Of The Product For Its Inventiveness:

Sugar-free hence it can be used extensively by diabetic patients.

- Improved bioavailability by formation of micro emulsion (Figure 1)

- Improved bioavailability by retarding the metabolism of Resveratrol.

It is evident from the following discussion that the present invention comprises a number of embodiments:

1. An oral aqueous pharmaceutical formulation characterized in that said formulation is sugar-free and comprises effective amounts of each of Resveratrol, Benfotiamine, and Pyridoxamine, or their pharmaceutically accepted salts, a bioavailability enhancer, and a hemodynamic and/or metabolic improvement compounds.

2. An oral aqueous pharmaceutical formulation as disclosed in embodiment 1, characterized in that said formulation has water as a vehicle, water miscible co-solvents and an effective amount of solubilizer, said solubilisers selected from a group Povidone and Polyoxyl 40,

hydrogenated castor oil, and the like.

3. An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 2, characterized in that said Polyoxyl 40 hydrogenated castor oil is present in an amount between 1% and 20% (w/w).

4. An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 3, characterized in that said Polyoxyl 40 hydrogenated castor oil is present in an amount between 1% and 5% (w/w). An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 4, characterized in that said Resveratrol is present in an amount between 20 mg/ml and 30 mg/ml (w/v).

An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 5, characterized in that said Povidone is present in an amount between 4% and 8% (w/w).

An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 6, characterized in that said Benfotiamine is present in an amount between 20 mg/ml and 30 mg/ml (w/v).

An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 7, characterized in that said Pyridoxamine is present in an amount between 20 mg/ml and 30 mg/ml (w/v).

An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 8, characterized in that said bioavailability enhancer is piperine provided in an amount between 0.001% and 0.100% (w/w).

An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 9, characterized in that said piperine is provided in an amount of about 0.05% (w/w).

An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 10, wherein said pharmaceutically accepted salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p- tolylsulfonic, citric, tartaric, methanesulfonic, and the like, and amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like.

12. An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 1 1, wherein said Resveratrol is derived from natural sources such as grape skins, wine, or other botanical sources such as P. cuspidatum or C. quinquangulata, or produced synthetically as 98% trans-Resveratrol, said trans-Resveratrol being trans-3,5,4'-trihydroxystilbene, 3,4',5-stilbenetriol, (E)- 5-(pOhydroxystyryl) resorcinol).

13. An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 12, wherein Benfoti amine is S-benzoylthi amine O-monophosphate. 14. An oral aqueous pharmaceutical formulation as disclosed in embodiments

1 to 13, wherein said hemodynamic and/or metabolic improvement compound is selected from a group comprising angiotensin converting enzyme inhibitors (ACE-I), angiotensin receptor blockers (ARB), beta- blockers, aldose reductase inhibitors, calcium blockers, diuretics, glycosaminoglycans, incretinmimetics, insulin, insulin sensitizers, statins, fibrates, glucose uptake inhibitors, sulfonylureas, superoxide dismutase (SOD) and SOD mimetics, thiamine pyrophosphate and its prodrugs, transketolase inhibitors, other AGE inhibitors that can mechanistically complement post-Amadori-inhibitors, and protein kinase C inhibitors, An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 13, wherein the one or more said compounds are selected from the group consisting of angiotensin converting enzyme inhibitors and angiotensin receptor blockers, or pharmaceutically acceptable salts thereof, in a pharmaceutically acceptable carrier. Non-limiting examples of angiotensin converting enzyme inhibitors for use in the present invention include benazepril, benazeprilat, captopril, delapril, fentiapril, fosinopril, libenzapril, moexipril, pentopril, perindopril, pivopril, quinapril, quinaprilat, ramipril, spirapril, spiraprilat, zofenopril, ceronapril, enalapril, indoiapril, lisinoprii, alacepril, and cilazapril, or pharmaceutically acceptable salts thereof.Non-limiting examples of angiotensin receptor blockers for use in the present invention include losartan, candesartan, irbesartan, olmesartan, valsartan, telmisartan, eprosartan, and tasosartan. An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 15, wherein said sweetener is selected from a group of sweeteners comprising neotame, mannitol, acesulfame potassium, aspartame, fructose, maltitol, xylitol, saccharin, sodium cyclamate, sucralose and sucrose, or a combination thereof and provided in an amount between 0.05% to 4%. An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 16, wherein said flavouring agent is selected from a group comprising natural and artificial flavors such as red grape flavour, tonovin, or mints (i.e., peppermint, etc.), menthol, chocolate, artificial chocolate, bubblegum, both artificial and natural fruit flavors (i.e., cherry, grape, orange, strawberry, etc.) or a combination thereof, and provided in an amount up to 5g per 100 mi of the formulation.

An oral aqueous pharmaceutical formulation as disclosed in embodiments 1 to 17, wherein said colouring agent is caramel and provided in an amo8nt between 0.01% to .2% w/v.

A process of making an oral aqueous pharmaceutical formulation, characterized in that the said process comprises the step of providing effective amounts of each of Resveratrol, Benfotiamine, and

Pyridoxamine, or their pharmaceutically accepted salts, a bioavailability enhancer, and a hemodynamic and/or metabolic improvement compound. A process of making an oral aqueous pharmaceutical formulation as disclosed in embodiment 19, characterized in that the said process comprises the steps of:

c. adding in sequence, a. a mixture of Resveratrol and PGL, b. Polyoxyl 40 hydrogenated caster oil, and c. glycerin, to said second tank and stirring it at 50 RPM to form a homogeneous solution, d. transferring the homogeneous solution of step c and purified water to a third tank and stirring the mixture at 600 RPM to form a homogeneous solution,

e. adding a mixture of purified water and sodium hydroxide stored in a fourth tank to a fifth tank and stirring it at 50 RPM to form a homogeneous solution,

f. adding in sequence a. a mixture of benfotiamine and water, b. a

mixture of PGL and Polyoxyl 40 hydrogenated caster oil, and c.

glycerine to said fifth tank and stirring the contents at 50 RPM to form a homogeneous solution,

g. transferring the mixture of step f along with purified water to said third tank and stirring the contents of said third tank and stirring the mixture at 600 RPM to form a homogeneous solution,

h. transferring the contents of said third tank to said first tank and stirring the contents of said first tank to form a homogeneous solution, i. transferring the contents of said first tank to a sixth tank and stirring the mixture at 900 RPM to form a homogeneous solution,

j. adding to said sixth tank in sequence, a. sorbitol solution (70%) N- crystalising, b. a mixture of purified water and sodium benzoate, c. a mixture of hot purified water and neotame, d. a mixture of purified water and citric acid monohydrate, and stirring the mixture at 400 RPM to form a homogeneous solution, k. further adding to said sixth tank in sequence a. mixture of purified water and pyridoxamine hydrochloride, b. a mixture of purified water, a first colour and a second colour, c. a mixture of purified water, FAC, manittol, and cyanocobalamin, d. a mixture of a first flavour, PGL, Polyoxyl 40 hydrogenated caster oil, and glycerin, and stirring the mixture at 900 RPM to form a homogeneous solution,

1. further adding to said sixth tank tonovin and stirring the contents of said sixth tank at 900 RPM to form a homogeneous solution, m. passing the contents of said sixth tank through a press filter and mix the filtered liquid with a final volume of purified water and stirring the mixture at 1000 RPM to form a homogeneous solution.

A method of treating diseases caused by formation of Advanced Glycation End Products or Advanced Lipoxidation End Products, characterized in that said method comprises a step of administering a pharmaceutical formulation comprising comprises effective amounts of each of Resveratrol, Benfotiamine, and Pyridoxamine, or their pharmaceutically accepted salts, a bioavailability enhancer, and a hemodynamic and/or metabolic improvement compounds.

A method as disclosed in embodiment 21, characterized in that said diseases comprise diseases and conditions including many degenerative diseases, such as diabetes, atherosclerosis, chronic renal failure, Alzheimer's and cancer. While the above description contains much specificity, these should not be construed as limitation in the scope of the invention, but rather as an

exemplification of the preferred embodiments thereof. It must be realized that modifications and variations are possible based on the disclosure given above without departing from the spirit and scope of the invention. Accordingly, the scope of the invention should be determined not by the embodiments illustrated, but by the appended claims and their legal equivalents.

Claims

An oral aqueous pharmaceutical formulation characterized in that said formulation is sugar-free and comprises effective amounts of each of Resveratrol, Benfotiamine, and Pyridoxamine, or their pharmaceutically accepted salts, a bioavailability enhancer, and a hemodynamic and/or metabolic improvement compound, a colouring agent, a flavoring agent, and a sweetener.

An oral aqueous pharmaceutical formulation as claimed in claim 1, characterized in that said formulation has water as a vehicle, water miscible co-solvents and an effective amount of solubilizer, said solubilisers selected from a group Povidone and Polyoxyl 40, hydrogenated castor oil, and the like.

An oral aqueous pharmaceutical formulation as claimed in claims 1 to 2, characterized in that said Polyoxyl 40 hydrogenated castor oil is present in an amount between 1% and 20% (w/w).

An oral aqueous pharmaceutical formulation as claimed in claims 1 to 3, characterized in that said Polyoxyl 40 hydrogenated castor oil is present in an amount between 1% and 5% (w/w).

An oral aqueous pharmaceutical formulation as claimed in claims 1 to 4, characterized in that said Resveratrol is present in an amount between 20 mg/ml and 30 mg/ml (w/v).

6. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 5, characterized in that said Povidone is present in an amount between 4%

7. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 6, characterized in that said Benfotiamine is present in an amount between 20 mg/ml and 30 mg/ml (w/v).

8. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 7, characterized in that said Pyridoxamine is present in an amount between 20 mg/ml and 30 mg/ml (w/v).

9. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 8, characterized in that said bioavailability enhancer is piperine provided in an amount between 0.001% and 0.100% (w/w).

10. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 9, characterized in that said piperine is provided in an amount of about 0,05%> (w/w).

11. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 10, wherein said pharmaceutically accepted salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohy drogencarbonic, phosphori c, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p- tolylsulfonic, citric, tartaric, methanesulfonic, and the like, and amino acids such as alginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like.

12. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 11, wherein said Resveratrol is derived from natural sources such as grape skins, wine, or other botanical sources such as P. cuspidatum or C. quinquangulata, or produced synthetically as 98% trans-Resveratrol, said trans-Resveratrol being trans-3,5,4'-trihydroxystilbene, 3,4',5-stilhenetrioL (E)- 5-(pOhydroxystyryl) resorcinol).

13. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 12, wherein Benfotiamine is S-benzoylthi amine O-monophosphate.

14. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 13, wherein said hemodynamic and/or metabolic improvement compound is selected from a group comprising angiotensin converting enzyme inhibitors (A.CE-I), angiotensin receptor blockers (ARB), beta-blockers, aldose reductase inhibitors, calcium blockers, diuretics, glycosaminoglycans, incretinmimetics, insulin, insulin sensitizers, statins, fibrates, glucose uptake inhibitors, sulfonylureas, superoxide dismutase (SOD) and SOD mimetics, thiamine pyrophosphate and its prodrugs, transketolase inhibitors, other AGE inhibitors that can mechanistically complement post-Amadori-inhibitors, and protein kinase C inhibitors.

15. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 13, wherein the one or more said compounds are selected from the group consisting of angiotensin converting enzyme inhibitors and angiotensin receptor blockers, or pharmaceutically acceptable salts thereof in a pharmaceutically acceptable carrier. Non-limiting examples of angiotensin converting enzyme inhibitors for use in the present invention include benazepril, benazeprilat, captoprii, delapril, fentiaprii, fosinoprii, libenzapril, moexipril, pentopril, perindopril, pivopril, quinapril, quinapril at, ramipril, spirapril, spiraprilat, zofenopril, ceronapril, enalapri l, indolapril, lisinopril, aiaceprii, and cilazapril, or pharmaceutically acceptable salts thereof.Non-limiting examples of angiotensin receptor blockers for use in the present invention include losartan, candesartan, irbesartan, olmesartan, valsartan, telmisartan, eprosartan, and tasosartan.

16. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 15, wherein said sweetener is selected from a group of sweeteners comprising neotame, mannitol, acesulfame potassium, aspartame, fructose, maltitol, xylitol, saccharin, sodium cyclamate, sucralose and sucrose, or a combination thereof and provided in an amount between 0.05% to 4%.

17. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 16, wherein said flavouring agent is selected from a group comprising natural and artificial flavors such as red grape flavour, tonovin, or mints (i.e. , peppermint, etc.), menthol, chocolate, artificial chocolate, bubblegum, both artificial and natural fruit flavors (i.e., cherry, grape, orange, strawberry, etc.) or a combination thereof, and provided in an amount up to 5g per 100 ml of the formulation.

18. An oral aqueous pharmaceutical formulation as claimed in claims 1 to 17, wherein said colouring agent is caramel and provided in an amo8nt between 0.01% to .2% w/v.

19. A process of making an oral aqueous pharmaceutical formulation, characterized in that the said process comprises the step of providing effective amounts of each of Resveratrol, Benfotiamine, and Pyridoxamine, or their pharmaceutically accepted salts, a bioavailability enhancer, and a hemodynamic and/or metabolic improvement compound.

20. A process of making an oral aqueous pharmaceutical formulation as claimed in claim 19, characterized in that the said process comprises the steps of:

n. mixing together purified water and Povidone by stirring at 50 RPM and adding the mixture to a first tank and stirring it to form a homogeneous solution,

o. mixing PGL and Piperine and adding the mixture to a to a second tank and stirring at 50 RPM to form a homogeneous solution,

p. adding in sequence, a. a mixture of Resveratrol and PGL, b. Polyoxyl

40 hydrogenated caster oil, and c. glycerin, to said second tank and stirring it at 50 RPM to form a homogeneous solution,

q. transferring the homogeneous solution of step c and purified water to a third tank and stirring the mixture at 600 RPM to form a homogeneous solution, r. adding a mixture of purified water and sodium hydroxide stored in a fourth tank to a fifth tank and stirring it at 50 RPM to form a homogeneous solution,

s. adding in sequence a. a mixture of benfotiamine and water, b. a mixture of PGL and Polyoxyl 40 hydrogenated caster oil, and c. glycerine to said fifth tank and stirring the contents at 50 RPM to form a homogeneous solution,

t. transferring the mixture of step f along with purified water to said third tank and stirring the contents of said third tank and stirring the mixture at 600 RPM to form a homogeneous solution,

u. transferring the contents of said third tank to said first tank and stirring the contents of said first tank to form a homogeneous solution, v. transferring the contents of said first tank to a sixth tank and stirring the mixture at 900 RPM to form a homogeneous solution,

w. adding to said sixth tank in sequence, a. sorbitol solution (70%) N- crystalising, b. a mixture of purified water and sodium benzoate, c. a mixture of hot purified water and neotame, d. a mixture of purified water and citric acid monohydrate, and stirring the mixture at 400 RPM to form a homogeneous solution,

x. further adding to said sixth tank in sequence a. mixture of purified water and pyridoxamine hydrochloride, b. a mixture of purified water, a first colour and a second colour, c. a mixture of purified water, FAC, manittol, and cyanocobalamin, d. a mixture of a first flavour, PGL, Polyoxyl 40 hydrogenated caster oil, and glycerin, and stirring the mixture at 900 RPM to form a homogeneous solution,

y. further adding to said sixth tank tonovin and stirring the contents of said sixth tank at 900 RPM to form a homogeneous solution, z. passing the contents of said sixth tank through a press filter and mix the filtered liquid with a final volume of purified water and stirring the mixture at 1000 RPM to form a homogeneous solution.

21. A method of treating diseases caused by formation of Advanced Glycation End Products or Advanced Lipoxidation End Products, characterized in that said method comprises a step of administering a pharmaceutical formulation comprising comprises effective amounts of each of Resveratrol, Benfotiamine, and Pyridoxamine, or their pharmaceutically accepted salts, a bioavailability enhancer, and a hemodynamic and/or metabolic improvement compounds.

22. A method as claimed in claim 21, characterized in that said diseases comprise diseases and conditions including many degenerative diseases, such as diabetes, atherosclerosis, chronic renal failure, Alzheimer's and cancer.