WO2011117894A1

WO2011117894A1 - Pharmaceutical technology of pharmaceutical composition in novel/sequential drug delivery system containing nitric oxide donor

Info

Publication number: WO2011117894A1
Application number: PCT/IN2011/000211
Authority: WO
Inventors: Jegannathan Srinivas
Original assignee: Kausalya, Srinivas
Priority date: 2010-03-26
Filing date: 2011-03-28
Publication date: 2011-09-29

Abstract

The present invention relates to pharmaceutical technology of pharmaceutical composition in Novel/Sequential Drug Delivery System containing Nitric Oxide Donors, more particularly, selective Endothelial Nitric Oxide Synthase Enhancers and in synergistic combination with Nitic Oxide Scavengers, Lipid Lowering Agents, as nanonized particles, from amongst, HMG Co-A Reductase Inhibitors, Ezetimibe and Fibrates and additionally other pharmaceutical substances that prevent/treat Atherosclerosis and provide round-the-clock cardio-protection. Following are the specific objectives of the present invention: a) Provide optimum benefits of Nitric Oxide and make it devoid of its adverse effects, through our unique technology and thus make it relatively safe for long-term usage. b) Provide synergistic combination of Lipid Lowering Agents with Nitric Oxide to achieve the dual objectives of a) Vasodilatation and b) Improve Dyslipidemia. c) Improve Quality of Life in patients with Ischemic Heart Disease.

Description

Title: Pharmaceutical technology of pharmaceutical composition in Novel/Sequential Drug Delivery System containing Nitric Oxide Donor. Technical Field

The invention pertains to pharmaceutical drug research that involves novel methods such as: a) Drugs in sequential release pattern and b) Drugs in nanonized form for improved bioabsorption and bioavailability.

BACKGROUND ART

The present invention relates to pharmaceutical technology of pharmaceutical composition in Novel/Sequential Drug Delivery System containing Nitric Oxide Donors, more particularly, selective Endothelial Nitric Oxide Synthase Enhancers and in synergistic combination with Nitic Oxide Scavengers, Lipid Lowering Agents, as nanonized particles, from amongst, HMG Co-A Reductase Inhibitors, Ezetimibe and Fibrates and additionally other pharmaceutical substances that prevent/treat Atherosclerosis and provide round-the-clock cardio-protection.

The relevant prior art methods, which teach adaptation of diverse delivery systems of the active, are as follows:

• Indian Patent Application No. : 1438/DELJ2005, DIRECTOR GENERAL, DRDO.

Nitric Oxide Donor based respiratory formulation for management of pulmonary hypertension and other hypoxic and a process for the preparation thereof.

• USPTO Patent Application 200801 9097 Beta-agonist compounds comprising nitric oxide donor groups and reactive oxygen species scavenger groups and their use in the treatment of respiratory disorders: The present invention relates to multifunctional β-agonist compounds that are capable of acting both as nitric oxide donors and as scavengers of reactive oxygen species and which are useful in the treatment of respiratory disorders. The invention further relates to methods of using such compounds in the treatment of respiratory disorders such as asthma.

• United States Patent 6103769.

The present invention concerns a pharmaceutical composition of a sterile, injectable physiologically acceptable solution containing nitric oxide (NO), a method for its preparation and a detection method for physiologically present NO. Physiologically present nitric oxide (NO), through a variety of biological functions, can influence essential key processes in the development of arteriosclerosis, such as relaxation of smooth vascular muscles, inhibition of adhesion of thrombocytes, granulocytes and monocytes to the vascular wall, inhibition of proliferation of secretory smooth muscle cells and direct effects on endothelial metabolism.

Nitroso Compounds or Nitric Oxide Releasing Compounds with Nitric Oxide Scavengers of Statins

• United States Patent 7166638

Statin nitroderivatives having improved pharmacological activity and enhanced tolerability are described. They can be employed for treating and/or preventing several diseases, in particular coronary syndromes, neurodegenerative disorders as well as for reducing cholesterol levels.

• US2005009888

This invention deals with the combination of the antihypertensive calcium channel blocker amlodipine and lipid- lowering agent atorvastatin inhibits free cholesterol crystallization in atherosclerotic-like membranes. In addition, it is claimed that treatment with a combination of amlodipine and atorvastatin results in a synergistic effect on the release of Nitric Oxide from aorta endothelial cells.

• Application No. : 4788/DELNP/2006, NOXILIZER, INC.

NITRIC OXIDE-RELEASING MOLECULE: This invention relates to compositions comprising carbon-based diazeniumdiolates that release nitric oxide (NO). The carbon- based diazeniumdiolated molecules release NO spontaneously under physiological conditions without subsequent nitrosamine formation. The present invention also relates to methods of preparing the carbon-based diazeniumdiolated molecules, compositions comprising such molecules, methods of using such compositions, and devices employing such molecule compositions.

• Application No. : 1759-1763/CHE/2006, MR. SRINIVAS J EGA NATHAN A therapeutic formulation comprising of sulfonylureas as active agents namely, Glibenclamide, Gliclazide, Glimepiride, Glipizide and Gliquidone, characterized in that said Sulfonylureas are combined with Metformin and combined or complexated with NO Scavenger to prevent Diabetic Complications arising out of Nitrosative stress.

• Indian Patent Application No. : 2856/DELNP/2004, THE UNIVERSITY OF QUEENSLAND

"METHODS AND COMPOSITONS COMPRISING NITRIC OXIDE DONORS AND OPIOID ANALGESICS" Compositions and methods that induce, promote or otherwise facilitate pain relief are disclosed. These compositions and methods comprise a nitric oxide donor which either directly or indirectly prevents, attenuates or reverses the development of reduced opioid sensitivity, together with a compound which activates the opioid receptor that is the subject of the reduced opioid sensitivity. The compositions and methods prevent or alleviate pain, especially in neuropathic conditions and even more especially in peripheral neuropathic conditions such as painful diabetic neuropathy (PDN). The preferred nitric oxide donor is L-arginine whilst the preferred compounds which activate the opioid receptor are morphine and oxycodone. Conjugate compounds comprising the nitric oxide donor and an opioid analgesic are also disclosed. · Indian Patent Application No. : 2496/MUMNP/2008, THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

COMPOSITIONS AND METHODS FOR THE DELIVERY OF NITRIC OXIDE

H-NOX proteins are mutated to exhibit improved or optimal kinetic and thermodynamic properties for blood gas NO delivery. The engineered H-NOX proteins comprise mutations that impart altered NO or 0<sub>2 </sub>ligand-binding relative to the corresponding wild-type H-NOX domain, and are operative as physiologically compatible mammalian blood NO gas carriers. The invention also provides pharmaceutical compositions, kits, and methods that use wild-type or mutant H-NOX proteins for the treatment of any condition for which delivery of NO is beneficial.

Pharmaco-therapeutic issues: Heart Disease is the N0.1 killer disease, with nearly 25% incidence, in the most productive age-group (25 to 69). As per the latest WHO Report, Heart Disease happens to be the No.1 killer disease world over and in India as well (almost 25%). It's reported that altered lipid profile (Dyslipidemia) is the most important risk factor and is normally corrected with Hypolipidemic Drugs such as Statins. Indeed, these drugs have certainly revolutionized the treatment of IHD (Ischemic Heart Disease), but may not be adequate enough by themselves to provide overall protection of the blood vessels. Physicians usually rely on Vasodilators such as Nitrates (NO), but are mostly restricted to short-term treatment to prevent acute attacks. Although NO (Nitric Oxide) has immense value in IHD, it's primarily used for short-term, since NO is regarded as the most potent Free Radical and none of the body's own antioxidant enzymes can scavenge them effectively.

All major risk factors for atherosclerosis such as hyperlipidemia, diabetes, hypertension, and smoking are associated with endothelial dysfunction. Impaired endothelial function has also been observed in animal models of atherosclerosis. Although the underlining mechanisms of endothelial dysfunction are multifactorial, the major cause is an impairment of the eNOS/Nitric Oxide pathway, which includes the reduced Nitric Oxide production by eNOS, increased degradation of Nitric Oxide by reaction with superoxide, and decreased sensitivity to Nitric Oxide¹. eNOS - Derived NO is an Anti-Atherosclerotic Principle¹

In the blood vessels, Nitric Oxide is produced from the endothelium mainly by the constitutively expressed eNOS, which is activated by shear-stress of the flowing blood or agonists such as bradykinin and acetylcholine. Besides its role as endothelium-derived relaxing factor (EDRF), Nitric Oxide protects blood vessels from thrombosis by inhibiting platelet aggregation and adhesion. In addition, endothelial Nitric Oxide possesses multiple anti-atherosclerotic properties, which include (i) prevention of leukocyte adhesion to vascular endothelium and leukocyte migration into the vascular wall; (ii) decreased endothelial permeability, reduced influx of lipoproteins into the vascular wall and inhibition of low-density lipoprotein (LDL) oxidation; and (iii) inhibition of DNA synthesis, mitogenesis, and proliferation of vascular smooth muscle cells.

Nitric Oxide is one of the few gaseous signaling molecules known and is additionally exceptional due to the fact that it is a radical gas. It is a key vertebrate biological messenger, playing a role in a variety of biological processes. Nitric oxide, known as the 'endothelium-derived relaxing factor', or 'EDRF', is biosynthesized endogenously from L-arginine, oxygen and NADPH by various nitric oxide synthase (NOS) enzymes. Reduction of inorganic nitrate may also serve to make nitric oxide. The endothelium (inner lining) of blood vessels uses nitric oxide to signal the surrounding smooth muscle to relax, thus resulting in vasodilation and increasing blood flow. Nitric oxide is highly reactive (having a lifetime of a few seconds), yet diffuses freely across membranes. These attributes make nitric oxide ideal for a transient paracrine (between adjacent cells) and autocrine (within a single cell) signaling molecule².

Studies have confirmed that non-selective Nitric Oxide Enhancers and Nitric Oxide Donors that spontaneously release Nitric Oxide can cause more harm than good due to the high likely-hood of potent oxidative damage with Nitric Oxide Free Radicals. Pharmaco-therapeutic solutions:

Thus researchers had been trying to find and investigate drugs that can selectively enhance Endothelial Nitric Oxide Synthases, and prevent the eNOS uncoupling. It's well known that eNOS uncoupling potentially causes release of dangerous peroxynitrite³.

Scientists have discovered number of small molecules that have the potential ^'to prevent eNOS uncoupling and, at the same time, enhance eNOS expression. These include the protein kinase C inhibitor midostaurin, the pentacyclic triterpenoids ursolic acid and betulinic acid, the eNOS enhancing compounds AVE9488 and AVE3085, and the polyphenolic phytoalexin trans-resveratrol. Such compounds enhance NO production from eNOS also under pathophysiological conditions and may thus have therapeutic potential⁴.

The oxidative stress in patients with IHD, Syndrome-X and Diabetes is generally quite high. The body has no known antioxidant enzyme that can scavenge deadly free radicals like Peroxynitrite, Nitric Oxide, etc. and thus the treatment warrants a good Nitric Oxide Scavenger, such as Rutin Hydrate. Cholesterol Reducers such as Statins have become the mainstay in preventing or treating atherogenic conditions. However, treating with Statins alone may not provide comprehensive endothelial protection. Patients with Hyperlipidemia do benefit a lot by taking Ezetemibe and Fibrates.

While treating patients with history of Ischemic Diseases or potential to develop the same, focusing only on Hyperlipidemia or Dyslipidemia may not provide the complete protection against atherogenicity and the related diseases. The main objective of our approach is to provide a holistic approach to vascular protection than addressing only the cholesterol related vascular risk. Uniqueness of this invention:

The present research is to alleviate the handicap in the Cardiologists' armamentarium and provide a comprehensive solution to treat IHD.

The novelty in the present invention is the unique approach of protecting the vascular endothelium through several pathways, more importantly in a manner that work synergistically with each other.

This invention relates to the field of Pharmaceutical Technology. More particularly the invention principally relates to pharmaceutical composition in Novel/Sequential Drug Delivery System containing Nitric Oxide Donors from amongst Midostaurin, Ursolic acid, Trans-Resveratrol and Betulinic acid, which enhance selective Endothelial Nitric Oxide Synthase (eNOS), without causing the deleterious oxidative damage of peroxynitrite (harmful free radicals), in synergistic combination with Nitric Oxide Scavengers,

Cholesterol Lowering Agents such as, HMG-CoA Reductase Inhibitors, (from amongst Atorvastatin, Rosuvastatin, Simvastatin, Lovastatin, Pravastatin, Pitavastatin, etc.), Selective Cholesterol Absorption Inhibitors (Ezetemibe) Fibrates and additionally pharmaceutical substances that promote vascular tone of the endothelium. The subject matter of the Patent Application relates to the technology/method of developing a pharmaceutical composition that can protect the endothelial function and promote the vascular tone of the endothelium.

The invention relates to providing additional active pharmaceutical ingredients that help to promote vascular tone of the endothelium, by providing synergistic action. The invention relates to the development of the pharmaceutical composition in a unique Sequential Drug Delivery System.

BRIEF DESCRIPTION OF DRAWINGS

In-lay Tablets/Multi-layered Tablets:

Physical/Chemical Separation: It is possible to avoid the incompatibility in between active-active; excipient-excipient and active-excipients by mean of physical separation. Well known example of such an interaction is a Millard reactions occurring during tablet compression.

Multiple release profile: Such drug delivery systems are able to provide multiple release kinetics of same/different drugs of same or different physicochemical properties by application of multiple layers. Each monolith was formulated in order to parcel out the delivery of drug dose by means of different release control mechanisms.

Drug Release Kinetic Analysis

Drug release kinetics were analyzed by the following phenomenological expression 1 1 :

M t M∞ = kt n (1 ) where M t , M∞ , k, and n are the amount of drug released at time t, the initial amount of drug in a tablet, the constant, and the release exponent, respectively. The exponent n shows the linearity of release kinetics. The first release data point has been excluded in this analysis to eliminate the effect of drug burst from the tablet surface.

If drug release kinetics are controlled solely by a surface erosion process without drug diffusion, the drug release rate may be expressed as follows: dM t dt = 2 π Lk e C o (x + y) (2) where k e , C o , x, and y are the erosion rate constant, the initial drug concentration in a tablet, the outer radius of a tablet, and the inner radius of a tablet, respectively. However, x + y = r o + r i according to Equation 3: surface area at t = 2 ττ L ( x + y ) = 2 π L{(r o - a) + (r i + a)} = 2 π L(r o + r i ) where a is the radially eroded thickness of the core tablet. Thus, Equation 2 becomes: dM t dt = 2 π Lk e C o (r o + r i ) (4) Integrating Equation 4 yields:

MtM°° = 2kero-rit (5) where M∞ = T LCo(ro2-ri2). Multilayered Buccal Tablets Multilayered buccal tablets would be of significant clinical value since the therapeutic effect can be immediate, which may be essential in most of the patients with IHD. A representative diagram of the multilayered tablet has been given below.

Layer

Transdermal Drug Delivery System

Polymers are the backbone of Transdermal Drug Delivery System, which control the release of the drug from the device. Polymer matrix can be prepared by dispersion of drug in liquid or solid state synthetic polymer base.

A schematic presentation of the delivery system given below: Transdermal Patch

OBJECT OF THE INVENTION: The main objective of this present invention is to provide pharmaceutical technology of delivering cardio-protective drugs in a pre-determined sequential release pattern for round- the-clock cardio-protection. The first objective of the present invention is to provide a pharmaceutical composition that provides Nitric Oxide Donor from amongst idostaurin, Ursolic acid, Trans-Resveratrol and Betulinic acid, which enhance selective Endothelial Nitric Oxide Synthase (eNOS), without causing the deleterious oxidative damage of peroxynitrite (harmful free radicals).

The second objective of this invention is to provide Nitric Oxide Scavengers, such as Rutin Hydrate to overcome the oxidative stress in vulnerable individuals.

The third objective of this invention is to provide Cholesterol Reducing Agents as nanonized particles, in synergistic combination with eNOS selective Nitric Oxide Donor. Another objective of the present invention is to provide HMG Co-A Reductase Inhibitor from amongst Atorvastatin, Rosuvastatin, Pitavastatin, Simvastatin, Lovastatin, etc., as nanonised particles.

Another objective of this invention is to provide Selective Cholesterol Absorption Inhitor like Ezetemibe, in synergistic combination with eNOS selective Nitric Oxide Donor and HMG Co-A Reductase Inhibitors.

Another objective of the present invention could be to provide Fibrates from amongst Gemfibrosil, Fenofibrate, Besafibrate, etc.

Yet another objective of the present invention is to provide a pharmaceutical composition that promotes vascular tone of the endothelium.

Further objective of the present invention is to provide the pharmaceutical composition in injectable dosage form. Yet another objective of the present invention is to provide the pharmaceutical composition in trans-dermal dosage form.

DESCRIPTION OF THE EMBODIMENTS In order to achieve the above-mentioned objective(s), we have invented novel technology that can prevent and treat Ischemic Heart Conditions like Atherogenicity, Dyslipidemia, and Endothelial Dysfunction and thereby achieve the objective of better quality of life in patients with Vascular Diseases

The composition(s) of the invention can be as follows

• Pharmaceutical composition containing eNOS selective Nitric Oxide Donor, for example, protein kinase C inhibitor midostaurin, the pentacyclic triterpenoids ursolic acid and betulinic acid and trans-resveratrol.

• The present invention can contain eNOS selective Nitric Oxide Donor in synergistic combination with Nitric Oxide

Scavengers such as, Rutin Hydrate, to scavenge deadly Nitric Oxide free radicals.

• The present invention can contain eNOS selective Nitric Oxide Donor in synergistic combination with Cholesterol

Reducing Agents, such as HMG Co-A Reductase inhibitors, Ezetimibe and Fibrates and additional active pharmaceutical ingredients to promote vascular tone of the endothelium

Thus the invention is an attempt to offer a solution for better quality of life in patients with Vascular Diseases due to Atherogenesis, Dyslipidemia and Endothelial Dysfunction. Examples:

As per the present invention, the outermost layer would contain eNOS selective Nitric Oxide Donor, for example, protein kinase C inhibitor midostaurin, the pentacyclic triterpenoids ursolic acid and betulinic acid and trans- resveratrol, in the appropriate doses for the synergistic action with Lipid Lowering Agents.

The second layer would contain Nitric Oxide Scavengers such as Rutin Hydrate for improved antioxidant activity.

The third layer would contain the specific Lipid Lowering Agents as nanonized particles, for improved bioavailability.

The third layer would also contain other anti-atherogenic substances that are part of this embodiment. Materials and Methods

The present invention can be provided as multilayered buccal tablets, wherein excipients could be selected from sodium carboxymethylcellulose, Carbopol-934 as Bioadhesive polymers to impart mucoadhesion. D-Mannitol and ethyl cellulose.

The present invention can also be provided controlled release method wherein the excipients could be selected from mannitol, ethyl cellulose, polyethylene glycol (particularly PEG 6000), HPMC and. HPMCAC.

The present invention can also be provided as Transdermal patches that contain natural or synthetic polymers or synthetic elastomers, with the drug and other components of the system such as penetration enhancers and Pressure Sensitive Adhesives. Market Potential

Heart Disease is the N0.1 killer disease, with nearly 25% incidence, in the most productive age-group (25 to 69). The major cause of death in such individuals is atherosclerosis, the diseased state of the arteries which results in heart attacks. The heart muscle like any other muscle requires an adequate blood supply, which is impeded in patients with atherosclerosis. Invariably, the arteries absorb mushy deposits of fat and cholesterol and gradually become blocked with a porridge-like substance called atheroma.

Besides, the arteries get hardened (sclerosis) and lose their tone and elasticity, leading to narrowing of the arteries

(vasoconstriction). The present invention is an attempt to achieve the dual objectives of cholesterol reduction with reliable and safe vasodilatation. Thus our innovation will be of immense value in crores of people with Heart Diseases, who may be vulnerable to Heart Attack. It would be our sincere effort to extend this benefit not only to the Indian market but also to the international markets.

The present research is to provide a comprehensive solution to patients diagnosed to be having Ischemic Heart Disease (IHD). The most important objective of our innovation is to improve the quality of life in patients with IHD by innovatively mitigating the long-term risk of acute attacks with a new comprehensive drug delivery package and sequenced drug delivery systems. The uniqueness in our final product would be that of a single simple solution to achieve the dual objectives of a) Lipid control and b) reliable vasodilatation with N.O. with reduced chances of its ill-effects. To achieve this objective, our invention is a Sequential Delivery System of N.O. Donor (selective to blood vessel), N.O. Scavenger and Hypolipidemic Drug(s), preferably in multi-layered tablets.

SUMMARY:

In view of the above circumstances, the composition of the present invention is in the form of a matrix tablet comprising the active ingredient, w-hydrophilic polymers, water-soluble and/or water dispersible diluents, pharmaceutically acceptable tablet excipients, and antibiotic adjuvant if any, for controlling the release of active ingredients. According to the present invention, the active ingredients are NO Donors, NO Scavengers and Cholesterol Reducers, or the pharmaceutically acceptable hydrates, salts or esters in a standard therapeutic dose, depending on the substance chosen References: 1. Current Pharmaceutical Design, 2009, 15, 3133-3145

2. Biochemistry, 4th Edition, pp.732

3. Byron's Wellness Resources

4. Curr Pharm Des. 2009; 15(27):3133-45.

5. Hobbs, et al. "Achievement of English National Service Framework Lipid-lowering Goals: Pooled Data from Recent Comparative Treatment Trials of ®atin at Starting Doses" J. Clin. Pract., vol. 59, 10 (2005), 1171- 1177.

Shishehbor, et al., "Statins Promote Potent Systemic Antioxidant Effects through Specific Inflammatory Pathways" Circulation. 2003; 108: 426-431. Rossiello et al., "Fibrin Down-regulates LPS- and PMA-induced Tissue Factor Expression by Blood Momonuclear Cells," Thromb Haemost 84 453-459, 2000.

Momi et al., "Prevention of Pulmonary Thromboembolism by NCX 4016, a Nitric Oxide-Releasing Asprin," European Journal of Pharmacology 397 177-185, 2000.

Emanueli et al., "Local Delivery of Human Tissue Kallikrein Gene Accelerates Spontaneous Angiogenesis in Mouse Model of Hindlimb Ischemia," Circulation 2/9 125-132, 2001.

Ongini et al., Nitric Oxide (NO)-Releasing Statin Derivatives, A Class of Drugs Showing Enhanced Antoproliferative And Antiinflammatory Properties, PNAS 101 8497-8502, 2004. Bonazzi et al., "New Nitric; Oxide (NO)-releasing 1111 Derivatives With Enhanced Anti-Inflammatory Properties," AHA Scientific Sessions 9-12, 2003.

Dever et al., "Effects of No-Pravastatin on Leukocyte Adhesion and Reactive Oxygen Species Generation in Control and Apoe Knockout Mice," AHA Scientific Sessions 9-12, 2003. Rossiello et al., "Nitropravastatin (NCX6550) Exerts an Antiplatelet/ Antithrombotic Activity and Inhabits Tissue Factor Expression," AHA Scientific Sessions 9-12, 2003. Emanueli et al., "The Nitric Oxide (NO)-Releasing Pravastatin Derivative, NCX 6550, Potentiates Reparative Angiogenesis In A Mouse Model Of Peripheral Ischemia," BPS, 2003. Guzzetta et al., "Nitric Oxide (NO)-Releasing Statin?: A New Class of Drugs Combining NO and jjOn Properties," BPS, 2003.

Momi et al., "Antiplatelet and Antithrombotic Activity of NCX 6550, A Nitric Oxide (NO)-Releasing Derivative of Pravastatin," BPS, 2003.

Guzzetta et al., "Novel Nitric Oxide (NO)-releasing Derivatives of Statins Modulate Cellular NO Homeostasis Acting on eNOS and iNOS Expressions," 9th International Conference on Alzheimer's Disease and Related Disorders, 2004.

• IN/PCT/2000/00518/ OL, "NITRIC OXIDE SYNTHASE INHIBITORS", GLAXO GROUP LTD

IN/PCT/2002/1520/CHE, "AMIDINO COMPOUND AND SALTS THEREOF USEFUL AS NITRIC OXIDE SYNTHASE INHIBITORS", PHARMACIA

CORPORATION

• Indian Patent No. 225619, PHARMACIA CORPORATION: 2-AMINO-2-ALKYL-5 HEPTENOIC

AND HEPTYNOIC ACID DERIVATIVES AS NITRIC OXIDE SYNTHASE INHIBITORS

Claims

CLAIMS We claim:

1. A Pharmaceutical technology of cardiovascular drugs released in sequential pattern for round-the-clock protection in conditions like IHD, Syndrome-X and Diabetes.

2. A pharmaceutical composition in Novel/Sequential Drug Delivery System containing Nitric Oxide

Donors - from amongst Midostaurin, Ursolic acid, Trans-Resveratrol and Betulinic acid, which enhance selective Endothelial Nitric Oxide Synthase (eNOS), without causing the deleterious oxidative damage of peroxynitrite (harmful free radicals).

3. A pharmaceutical composition as per Claim 2 in synergistic combination with Nitric Oxide Scavengers such as| Rutin Hydrate to reduce oxidative.

4. A pharmaceutical composition as per Claim 2 and Claim 3 in synergistic combination with HMG-CoA Reductase Inhibitors as nanonised particles from amongst Atorvastatin, Rosuvastatin, Simvastatin, Lovastatin, Pravastatin, Fluvastatin, Pravastatin, Mevastatin and any other identical analogue.

5. A pharmaceutical composition as per Claim 2 and Claim 3 in synergistic combination with Ezetemibe or any other selective cholesterol absorption inhibitor.

6. A pharmaceutical composition as per Claim 2 and Claim 3 in synergistic combination with Fibrates from amongst Gemfibrosi!, Besafibrate, Fenofibrate, Clofibrate, Simfibrate and Ciprofibrate.

7. A pharmaceutical composition as per Claim 2 and Claim 3 in synergistic combination with cardioprotective nutrients, Folic acid and Vitamin D.

8. The Pharmaceutical composition as per Claim 2, Claim 3 in synergistic combination with composition as per Claim 4 to Claim 7 to be provided multilayered buccal delivery system and multilayered controlled delivery system.

9. The Pharmaceutical composition as per Claim 2, Claim 3 in synergistic combination with composition as per Claim 4 to Claim 7 to be provided in injectable dosage form.

10. The Pharmaceutical composition as per Claim 2, Claim 3 in synergistic combination with composition as per Claim 4 to Claim 7 to be provided in trans-dermal dosage form.