Polyherbal composition for optimum vascular activity and cardiac health

Abstract

The present invention relates to the polyherbal composition for optimum vascular activity and cardiac health. The Invention also provides its method of preparation. The composition exhibits beneficial effects in regulating dyslipidaemia, controlling and reducing arterial fat deposition along with reducing elevated levels of serum cholesterol, triglycerides and associated atherosclerosis. The composition also regulates lipid levels and promotes cardiovascular health, as well as improves overall cardiac function.

Description

[0001]B5156-00007 POLYHERBAL COMPOSITION FOR OPTIMUM VASCULAR ACTIVITY AND CARDIAC HEALTH FIELD OF INVENTION: The present invention relates to the polyherbal composition for optimum vascular activity and cardiac health. The Invention also provides its method of preparation. BACKGROUND OF THE INVENTION: The following background discussion includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication expressly or implicitly referenced is prior art. Atherosclerosis refers to the thickening / narrowing of blood vessels, resulting in reduction in blood flow and flexibility. This condition can lead to coronary atherosclerotic heart disease, also known as ischemic heart disease, where the heart experiences inadequate blood supply due to hardened coronary arteries. Symptoms such as chest pain (anginal syndrome), myocardial infarction, irregular heartbeat, and heart failure may or may not occur in association with coronary atherosclerosis. According to research from various sources, coronary heart disease is commonly attributed to the buildup of fatty deposits in the coronary artery and blood platelet aggregation in the connective tissues of these arteries. This accumulation leads to the development of atherosclerotic plaque, ultimately restricting blood flow to the heart and other organs. Additionally, smooth muscle spasms can contribute to the onset of coronary heart disease and stroke. Risk factors such as, high levels of total cholesterol and low-density lipoprotein (LDL), along with hypertension, smoking and work-related stress also significantly increases susceptibility for developing coronary heart disease. Atherosclerosis is a complex genetic disease in mammals marked by the build-up / deposition of lipid plaques in the walls of arteries such as the aorta, coronary arteries and carotid. As the condition advances these plaques can become calcified to varying degrees and are linked to the accumulation of cholesterol esters mainly in fatty deposits within the arteries. As cholesterol gathers in foam cells within arterial walls, it leads to narrowing of blood vessels and reduced blood flow. This process also involves thickening of arterial walls through smooth muscle hypertrophy, foam cell formation, and fibrous tissue buildup. Consequently, high levels of B5156-00007 cholesterol can lead to severe cardiovascular conditions like peripheral vascular disease, circulatory failure, cerebral vascular accidents, heart attack, and stroke with sudden death. Hypertension is a significant risk factor for atherosclerosis. Studies have shown that individuals with hypertension are at a higher risk of developing atherosclerosis, mainly because of high blood pressure increases the pressure on the arterial walls, leading to structural changes that predispose to atherosclerosis. Additionally, hypertension may promote the expression of several pro-inflammatory molecules, increasing oxidative stress and aggravate further atherosclerosis. Coronary atherosclerotic heart disease is typically managed with a vasodilator to widen the blood vessels and clear obstructions in the coronary arteries. Nonetheless, this approach is linked to numerous adverse effects including flushed face, headache, dizziness, nausea, fatigue, and specific allergic responses. In more serious instances, coronary bypass surgery or angioplasty may be employed / required. In the field of pharmaceutical chemistry, it is essential to note that it involves substances like the one listed below to control these issues. Statins are recognized for their ability to decrease the production of LDL in the liver by obstructing HMG CoA reductase. Despite this, they have been linked to liver damage and muscle tissue injury. Furthermore, they only contribute a modest reduction of up to 5% in atheromatous plaque thickness after an 18-month course of treatment and come with a relatively high cost. Niacin: This substance reduces LVDL in the liver, leading to a decrease in blood triglyceride levels. However, it can cause severe side effects such as intense itching, skin rash, and may also result in gastric ulcer and liver damage. The new delayed-release formulations of this compound are costly. Angiotensin Converting Enzyme ACE Inhibitors: ACE inhibitors helps in reducing the vascular resistance and thus reduced cardiac load, which is helpful not only reducing the blood pressure but also reduces the chances of stroke and myocardial infarction. Aspirin: It is a known inhibitor of plaque aggregation but has the potential to cause ulcers and bleeding in the digestive tract, particularly among susceptible individuals or with prolonged use. B5156-00007 Clopidogrel: Like many other compounds used to address plaque aggregation, this medication is expensive and can lead to adverse effects. Ezetimibe inhibits the intestinal absorption of cholesterol but does not directly reduce plaque. Challenges with these medications are affordability and frequent adverse effects. Fibrates activate lipoprotein lipase, which breaks down VLDL and chylomicrons in the liver, releasing their lipid components for HDL synthesis. However, they also carry a risk of hepatotoxicity and are costly. These medications, whether used alone or in various combinations for treating different factors and mechanisms that contribute to arterial disease, have not demonstrated significant reversal of artery blockage as claimed by the current invention. This is attributed to the synergistic effect produced by a combination of different herbal extracts with minimal (no or negligible) side effects and (comparatively) low cost. Hence, there is a need of an effective herbal / natural composition and its method of preparation for Optimum Vascular Activity and Cardiac Health with minimal or no side effects. OBJECTIVE OF THE INVENTION: The primary objective of the present invention is to overcome the drawbacks associated with prior art. An object of the present invention is to provide a safe efficacious poly herbal multicomponent formulation. Another object of the present invention is to provide a composition which exhibits beneficial effects in regulating dyslipidaemia and controlling arterial fat deposition. Another object of the present invention is to provide a composition which exhibits efficacy in reducing elevated levels of serum cholesterol, triglycerides and associated conditions such as atherosclerosis. Another object of the present invention is to provide a composition which regulates lipid levels and promoting cardiovascular health, as well as improving cardiac function. Another object of the present invention is to provide a composition which regulates blood pressure levels and promotes overall cardiovascular health. B5156-00007 SUMMARY OF THE INVENTION: In an aspect the present invention provides a polyherbal composition for improving Vascular Activity and Cardiac Health, comprising: a) Zingiber officinale present in an amount ranging from 0.25 – 5.5% b) Allium sativum present in an amount ranging from 0.15- 4.75% c) Citrus limon present in an amount ranging from 0.1- 3.75% d) Terminalia arjuna present in an amount ranging from 0.3-5.2 % e) Vitis vinifera present in an amount ranging from 0.2-4.5 % f) Moringa oleifera present in an amount ranging from 0.3-5.0 % g) Punica granatum present in an amount ranging from 0.25-3.6 % h) Ammannia auriculata present in an amount ranging from 0.2-4.4 % i) Alnus nepalensis present in an amount ranging from 0.5-5.5 % j) Atalantia monophylla present in an amount ranging from 0.5 – 2.5 % k) Apium graveolens present in an amount ranging from 1.2 -4.4 % l) Saccharum officinarum present in an amount ranging from 0.4-3.0 %. The Invention also provides a process of preparing the poly herbal composition for improving Vascular Activity and Cardiac Health, comprising the steps of: a) Drying the plant material as described above, at 45-55°C till its moisture content reaches <8%; b) Milling the dried plant material and passing through mesh size 60, for extraction through pulverization, to obtain a mixture; c) Boiling the mixture in 5-10 fold amount of water 3-6 times for 30 minutes each, followed by fermenting the mixture to 24-36 hours at 30 to 350C; d) Incubating the mixture obtained in step (c) with microbes Lactobacillus spp. having CFU in the range of 2 × 104to 3 × 106and transferring the mixture into a vessel followed by fermenting the mixture to 24-36 hours at 30 to 350C, to obtain a solution; B5156-00007 e) Filtering the solution to remove fine particles followed by heating for 30 min at 60-90°C to obtain a solution; f) Concentration the solution obtained in the above step under low temperature to extract until its relative density reaches from 1.1 to 1.4; g) Mixing the concentrated extract in a blender for 15-30 min with lemon and pomegranate Juice or a mixture thereof present in an amount of 0.5 to 2.5% based on the total weight of plant material. The Invention also provides a process of preparing the polyherbal nano composition as described above for improving Vascular Activity and Cardiac Health comprising and excipient / s selected from the components as mentioned below: D-α-Tocopherol polyethylene glycol 1000 succinate, Copolyvidone, Propylene glycol monocaprylate, polyethylene glycol, lecithin and Polyoxyethylene lauryl ether. The process of preparing the polyherbal nano formulation comprises the below steps: a) Mixing the polyherbal formulation as described above along with the excipients selected from D-α-Tocopherol polyethylene glycol 1000 succinate present in an amount ranging from 0.01 - 0.02%, Copolyvidone present in an amount ranging from 0.20 - 2.00%, Propylene glycol monocaprylate present in an amount ranging from 0.50 - 2.00%, polyethylene glycol present in an amount ranging from 0.75 - 2.00%, lecithin present in an amount ranging from 0.10 - 5.00% and Polyoxyethylene lauryl ether present in an amount ranging from 0.01 - 0.10%; b) Sonicating the admixture for 25 to 30 minutes in a probe sonicator at an amplitude ranging between 20 to 45% and temperature between 25 to 45°C, to obtain a nano- sized admixture; c) Homogenising the nano-sized admixture for 5-10 minutes at 10,000 to 15,000 RPM, to obtain a synergistic nano herbal composition. DETAILED DESCRIPTION OF DRAWINGS: To further clarify advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which is illustrated in the appended drawings. It is appreciated that these drawings depict only B5156-00007 typical embodiments of the invention and are therefore not to be considered limiting of their scope. The invention will be described and explained with additional specificity and detail with the accompanying drawings in which: Fig.1: Illustrate process of the present invention, according to an embodiment. Fig.2: Illustrate Particle size distribution obtained in Composition 24213 using Process A Fig.3: Illustrate Particle size distribution obtained in Composition 24213 using Process B Fig.4: Illustrate changes in levels of Cholesterol in study 1 Fig.5: Illustrate change in levels of Triglycerides in study 1 Fig.6: Illustrate change in levels of Cholesterol in study 2 Fig.7: Illustrate change in levelsof Triglycerides in study 2 Fig.8: Illustrate histopathological changes in study 2 Fig.9A: Illustrate Histopathology of Disease Control Group Animal Aorta Showing normal tunica interna, media and externa and infiltration of adipose tissue (arrow) Fig.9B: Illustrate Histopathology of Composition 24213 Group G4 Group Animal Aorta Showing normal tunica interna, media and externa. DETAILED DESCRIPTION: For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated system, and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur to one skilled in the art to which the invention relates. It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be restrictive thereof. The terms “comprises”, “comprising”, “includes”, or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a setup, device or method that comprises a list of components or steps does not include only those components or steps but may include other B5156-00007 components or steps not expressly listed or inherent to such setup or device or method. In other words, one or more elements in a system or apparatus proceeded by “comprises... a” does not, without more constraints, preclude the existence of other elements or additional elements in the system or method. The Invention provides a safe, efficacious herbal multicomponent formulation, developed by synergistic integration of various plant extracts in specific manner. The composition has been tested for its effectiveness through various studies to evaluate antioxidant, lipase inhibition, angiotensin-converting enzyme (ACE) inhibition potential. The efficacy of the composition was also validated using well-established animal models of hyperlipidaemia and associated atherosclerosis. The results demonstrated that Composition exhibits beneficial effects in regulating dyslipidaemia and controlling arterial fat accumulation. The composition demonstrates various advantages like: • Efficacy in reducing elevated levels of serum cholesterol, triglycerides and associated atherosclerosis. • Regulating lipid levels and promoting cardiovascular health, as well as improving cardiac function. • Helps to regulate blood pressure levels. • Promotes overall cardiovascular health. In an embodiment, the method to prepare the composition of the present invention comprises following steps: • All the collected samples of plants were cleaned (Removal of Unwanted / foreign material followed by washing) and dried in tray drier at 45-55°C till their moisture content reaches <8%; • Plant bark and seeds were milled and passed through sieve (mesh size 60) to increase their surface area for extraction using commercial pulverizer; • Mixture was boiled in 5-10 fold amount of water 3-6 times for 30 minutes each and the marc was combined. Combined marc was inoculated with microbes Lactobacillus spp. having CFU in the range of 2 × 104to 3 × 106. Following inoculation, the solution was transferred to a vessel and allowed to ferment 24-36 hours at 30-35°C. B5156-00007 • Prepared content was filtered using suitable filtration methodology to remove fine particles followed by heating for 30 min at 60-90°C. • Prepared liquid was concentrated using concentrator under low temperature to extract until its relative density reaches 1.1 to 1.4; the prepared admixture was named as extract 42. • Concentrated extract were mixed in a blender 15-30 min with lemon and pomegranate Juice or a mixture thereof (Mixture 2) in an amount of 0.5 to 2.5% based on the total weight of extract 42, for obtaining a mixture which is named as Composition 24213, in the present specification. The free radical scavenging activity was measured in terms of hydrogen donating or radical scavenging ability using the stable radical DPPH.200 µM solution of DPPH in methanol was prepared and 150µl of this solution was added to 50 µl of different test compounds at different concentrations. After 30 minutes, the absorbance was measured at 517 nm. Absorbance of control – Absorbance of test% Scavenging =X 100 Absorbance of control After studying the antioxidant activity of individual herbal extract, different compositions were prepared as under and their antioxidant activity was evaluated. B5156-00007 Composition 1 (Composition 24211) Composition 2 (Composition 24212) Composition 3 (Composition 24213) B5156-00007 Further Screening of developed compositions was carried out using methods such as Lipase inhibition action and Angiotensin convertase enzyme (ACE) inhibitory action. Lipase Assay Lipase inhibition activity was carried out using Porcine Pancreatic Lipase of the selected test compounds by the method described by Chedda et al. PNPB (para-nitrophenylbutyrate) was used as Substrate. 25µl of Test sample / Positive Control / Vehicle was incubated with 50µl of Enzyme solution (Lipase) and 100 µl phosphate buffer (pH 7.2) for 10 min at 37°C. After completion of Incubation 50µl of substrate was added to initiate the reaction. Change in absorbance was measured using Biotek multimode reader at 400 nm. The % inhibition = (1 –A / Ao) x 100 Where A is the absorbance of control and Aois the absorbance of sample. The comparative analysis of lipase inhibition by the three compositions reveals a clear hierarchy in their efficacy. Composition 3 (Composition 24213) stands out as the most potent formulation, with the lowest average IC50 and the highest consistency in its inhibitory effect. These findings indicate significant implications for the development of formulation aimed at controlling lipid metabolism. ACE inhibition Assay ACE activity was assayed by monitoring the release of HA from the substrate HHL. The assay mixture contained 35µl of 0.05 M sodium borate buffer of pH 8.2 containing 0.3 M NaCl, and 35 µl of test sample then add 10µl of ACE enzyme (1miliUnit) incubate for 10 min. at 37°C. Later add 20µl of 5 mM HHL again incubate at 37°C for 30 min. The reaction was arrested by the addition of 50 µl of 1 M HCl. Later Pyridine 100µl was added followed by 50µl of BSC and mixed by inversion for 1 min and cooled on ice. The yellow colour developed was measured at 410 nm in a spectrophotometer. B5156-00007 ^^^^^^^^^^^^^^^^^^^^ ^^^^ ^^^^^^^^^^^^^^ − ^^^^^^^^^^^^^^^^^^^^ ^^^^ ^^^^^^^^% ^^^^ℎ^^^^^^^^^^^^^^ ^^^^^^^^^^ =^^ 100 ^^^^^^^^^^^^^^^^^^^^ ^^^^ ^^^^^^^^^^^^^^ Composition 3 (Composition 24213) emerged as the most effective inhibitor, followed by Composition 2 (Composition 24212), and then Composition 1 (Composition 24211). The nano formulation was prepared for composition 24213, through the process as mentioned below: NANO-FORMULATION: Process A: The solubility and compatibility of Composition 24213 were assessed using various excipients such as D-α-Tocopherol polyethylene glycol 1000 succinate, Copolyvidone, Propylene glycol monocaprylate, and Poloxamer 124, in water to ensure optimal performance. The finalised formulation was then subjected to sonication for 15 to 35 minutes in a probe sonicator at an amplitude of 20-40%, resulting in a nano-sized admixture. This nano-sized admixture was subsequently homogenised for 3 to 5 minutes at 8,000 to 9,000 RPM, thereby forming final nano-emulsion. Process B The solubility and compatibility of Composition 24213 were evaluated using different excipients, including D-α-Tocopherol polyethylene glycol 1000 succinate, Copolyvidone, Propylene glycol monocaprylate, polyethylene glycol, lecithin and Polyoxyethylene lauryl ether. The admixture was sonicated for 25 to 30 minutes in a probe sonicator at anamplitude ranging between 20 to 45% and a temperature between 25 to 45°C, resulting in a nano-sized admixture. The nano-sized admixture was then homogenised for 5-10 minutes at 10,000 to 15,000 RPM, forming a synergistic nano herbal composition ready for use The nanoparticles synthesized through Process A exhibited a broader size distribution, with an particle diameter in the range of 925 - 1250 nm. This considerable variance in size suggests B5156-00007 that Process A may be less consistent in controlling the stages of particle formation. Conversely, Process B demonstrated remarkable precision in nanoparticle generation. The average particle size was significantly smaller and more uniform, measuring in the range of upto 200 nm. Moreover, the standard deviation associated with Process B was notably lower, indicating a higher level of reproducibility and control over the particle synthesis process. In conclusion, Process B stands out as the superior method for preparation of nanoparticles with optimum size achievement and minimal deviation. PREPARATION OF PHARMACEUTICAL COMPOSITION: In order to make a pharmaceutical composition acceptable, excipients which were selected from a range of additives, including gums, sweeteners, coatings, binders, lubricants, disintegration agents, suspending agents, granulating agents, solvents, colorants, glidants, anti-adherents, anti-static agents, surfactants, plasticizers, emulsifying agents, flavouring agents, viscosity enhancers were added to Compositions. Thus developed formula can be used to prepare various suitable dosage forms, including powders, pastes, granules, capsules, tablets, liquids, lozenges, emulsions, suspensions, syrups, elixirs, oral drops, jellies, phytopharmaceuticals, food supplements, and nutraceuticals. In silico SCREENING OF SELECTED INGREDIENTS FROM COMPOSITION 24213: Inhibiting key enzymes and receptors in the cardiovascular system is essential for treating cardiovascular disorders. APOB1 inhibition reduces LDL cholesterol, lowering atherosclerosis risk. Angiotensin II is targeted by ACE inhibitors and ARBs to manage hypertension and heart failure. β-blockers inhibit β-adrenergic receptors to control heart rate and reduce cardiac workload. MAPK pathway inhibitors help prevent pathological cardiac remodelling and hypertrophy. Statins inhibit HMG-CoA reductase, significantly reducing LDL cholesterol and preventing coronary artery disease. PPAR-γ modulation improves insulin sensitivity and reduces inflammation, benefiting cardiovascular health. Method: The ligands such as APOB1, Angiotensin, HMG-COA, β-adrenergic receptors; MAPK and PPAR-γ were selected due to their involvement in CVS / Hypertension. Three- dimensional structures of these proteins were obtained from the PDB databases which were than processed to generate 3D structure with least energy after removal of heteroatoms etc. Active site identification was carried out using online server Deepsite. ADMET and Drug B5156-00007 likeness studies were carried out pkCSM tool. The ligand and receptor interaction were done by using the Autodock 4.5.6 tool which are summarized in table below B5156-00007 * mM#nM Docking with multiple ligands can predict synergistic or antagonistic effects between ligands that can bind to the same receptor. This approach will be also helpful in deciphering the rational design of combination therapies or multicomponent formulations, in order to achieve synergistic effects or target multiple pathways involved in complex diseases. Table below is showing the fold change in inhibition constant after multiple ligands docking The combination of SDG, Apigenin, and Piperine with APOB1 resulted in a significant decrease in binding energy, suggesting a strong synergistic effect between these compounds when interacting with the protein. The concentration of this multi-compound complex was notably lower at 26.17 µM in comparison to 12.16 mM. The interaction of Angiotensin with Chlorogenic acid and Allicin, both individually and in combination, presents intriguing possibilities for modulating cardiovascular function. B5156-00007 Chlorogenic acid appears to have a significant impact on Angiotensin's binding affinity, which could be harnessed in therapeutic applications. Combination of Kaempferol and 6-Gingerol showed a synergistic effect, with an enhanced binding energy of -6.71 kcal / mol and a significantly lower Ki of 9.99 µM. This suggests that when used together, these compounds may offer a more potent inhibitory effect on HMG-CoA reductase. Data from in-silico studies showed that selected phytochemicals have interaction with the ligands / enzymes which plays important role in Fat metabolism and CVS management. They also showed 1.25 to 464.6 fold changes in the inhibition constant. QUALITY CONTROL PARAMETERS OF DEVELOPED FORMULATION Quantification of secondary compounds was conducted for the Composition 24213 to ensure consistency and potency across different batches, thus aiding in achieving therapeutic goals. IN VIVO EFFICACY STUDIES Effect of Developed herbal formulation on management of Cholesterol and Triglyceride levels were carried out using established animal models. STUDY 1 (TRITON INDUCED MODEL OF HYPERLIPIDEMIA) Thirty Two male Wistar rats were divided in to four groups of eight each. The rats from groups 1 and 2 received water once a day orally at a dose of 10 ml / kg b.w. and were served as normal and disease controls respectively. Rats from groups 3 received Chemical control at a dose of 30 mg / kg, b.w. While the animals of Group 4 treated with Composition 24213 (3 ml / kg b.w.). All the rats received the respective assigned treatment for 14 days. On 15th day, All the animals except group 1 was treated with Triton WR 1339 emulsion at a dose of 250mg / kg via i.p. injection, after 24 hours the blood was collected from the retro-orbital plexus under mild anaesthesia and subjected for the estimation of Lipid markers. B5156-00007 N=8 animals per group STUDY 2 (TRITON INDUCED MODEL OF HYPERLIPIDEMIA) Thirty Two male Wistar rats were divided in to four groups of eight each. The rats from groups 1 and 2 received water once a day orally at a dose of 10 ml / kg b.w. and were served as normal and disease controls respectively. Rats from groups 3 received Positive control at a dose of 30 mg / kg, b.w. While the animals of Group 4 treated with Composition 24213 (3 ml / kg b.w.). All the rats received the respective assigned treatment for 21 days. On 21stday, All the animals except group 1 was treated with Triton X-100 emulsion at a dose of 100mg / kg via i.p. injection, after 24 hours the blood was collected from the retro-orbital plexus under mild anaesthesia and subjected for the estimation of Lipid markers. N=8 animals per group Microscopic examination of aorta from animal’s studies showed accumulation of Perivascular adipose tissue in Disease control group when compared with normal control group. Noticeable atheromatous thickening in the intima of the aorta was not observed in Composition 24213 and Chemical control treated animals and composition 24213 was at par with the chemical control agent. Vascular edema was also reduced in the both the treatments groups i.e. Chemical control and Composition 24213. B5156-00007 Also in the efficacy studies and Safety studies carried out in rodents for Composition 24213, it was observed that the formulation was safe for oral administration and no toxicity was observed in animals when administered over a period of 3 months. Case Studies of effect of Composition 24213 Case 1: A 57-year-old male patient with Ischemic Heart Disease experienced significant improvements after using Composition 24213 as supportive medicine alongside standard conservative management. The patient's Left Ventricular Ejection Fraction (LVEF) increased from 45%-50% to more than 55%, diastolic function normalized from Grade 2 dysfunction, and symptoms of palpitation, breathlessness, and dyspnea were notably reduced. These results suggest that the Composition 24213 may effectively enhance symptom relief and cardiac function in ischemic heart disease patients. Case 2: A 76-year-old male patient with Ischemic Heart Disease and chronic diabetes experienced significant improvements after using Composition 24213 as adjuvant therapy alongside standard treatment. His Left Ventricular Ejection Fraction (LVEF) improved from 25%-30% to 45%-50%, and his heart pumping function showed significant enhancement. Additionally, symptoms such as palpitation, breathlessness, and chest discomfort were notably reduced. Case 3: The patient's stroke volume increased from 35.8 mL to 37.61 mL, and LV regurgitation fraction decreased significantly from 22.46 to 4.26. Pulmonary vascular resistance was reduced from 442 dyn·s·cm⁻⁵ to 50 dyn·s·cm⁻⁵, while pulmonary capillary pressure decreased from 18.96 mmHg to 2.6 mmHg. Measured arterial compliance improved from 0.64 mL / mmHg to 1.163 mL / mmHg, and mean systemic blood flow increased from 2.14 mL / min / 100g to 5.32 mL / min / 100g. The mitral valve orifice area expanded from 1.39 cm² to 4.3 cm², though the aortic valve orifice area observed a slight decrease from 3.1 cm² to 2.94 cm². Coronary flow reserve increased from 2.42 to 4.37, and fluid retention dropped from 30ml to 3ml. The global blood flow deficiency index improved significantly from 13 to 0, and LV ejection fraction increased from 22 % / s to 41 % / s. Case 4: The patient's treatment resulted in several significant improvements. Pulmonary capillary pressure decreased from 9.86 mmHg to 2.44 mmHg, and fluid retention dropped significantly from 60 ml to 2 ml. Pulmonary vascular resistance was reduced from 231 dyn·s·cm⁻⁵ to 67 dyn·s·cm⁻⁵, and LV regurgitation fraction decreased from 8.61% / s to 4.11 % / s. Although measured arterial compliance slightly declined from 0.354 mL / mmHg to 0.265 B5156-00007 mL / mmHg, mean systemic blood flow saw a minor reduction from 5.5 mL / min / 100g to 4.81 mL / min / 100g. There was a notable increase in the mitral valve orifice area from 2.3 cm² to 3.83 cm², while the aortic valve orifice area decreased slightly from 2.87 cm² to 2.38 cm². Coronary flow reserve significantly improved from 1.7 to 7.1, and end-systolic volume increased from 0.75 ml to 8.11 ml. additionally, the minimum LV ejection fraction rose from 44% to 63%. Case 5: The patient showed significant improvements across various cardiovascular parameters following treatment. Stroke volume increased markedly from 28.32 mL to 51.62 mL, and cardiac output rose from 2.22 to 3.54. Pulmonary vascular resistance decreased considerably from 174 dyn·s·cm⁻⁵ to 121 dyn·s·cm⁻⁵, while the LV ejection rate improved from 66 % / s to 102 % / s. Measured arterial compliance saw a notable rise from 0.764 mL / mmHg to 1.357 mL / mmHg, and mean systemic blood flow increased from 3.42 mL / min / 100g to 5.67 mL / min / 100g. The mitral valve orifice area expanded from 2.34 cm² to 3.49 cm², and the aortic valve orifice area increased significantly from 2.7 cm² to 4.35 cm². Additionally, the minimum LV ejection fraction improved from 26 % to 37 %, and the maximum LV ejection fraction improved significantly from 36% to 63%.

Claims

B5156-00007 We Claim:

1. A poly herbal nano composition improving Vascular Activity and Cardiac Health, comprising: a) Zingiber officinale present in an amount ranging from 0.25 – 5.5% b) Allium sativum present in an amount ranging from 0.15- 4.75% c) Citrus limon present in an amount ranging from 0.1- 3.75% d) Terminalia arjuna present in an amount ranging from 0.3-5.2 % e) Vitis vinifera present in an amount ranging from 0.2-4.5 % f) Moringa oleifera present in an amount ranging from 0.3-5.0 % g) Punica granatum present in an amount ranging from 0.25-3.6 % h) Ammannia auriculata present in an amount ranging from 0.2-4.4 % i) Alnus nepalensis present in an amount ranging from 0.5-5.5 % j) Atalantia monophylla present in an amount ranging from 0.5 – 2.5 % k) Apium graveolens present in an amount ranging from 1.2 -4.4 % l) Saccharum officinarum present in an amount ranging from 0.4-3.0 %.

2. A process for preparation of herbal composition improving Vascular Activity and Cardiac Health as claimed in claim 1, comprising: a) Drying the plant material as claimed in claim 1, at 45-55°C till its moisture content reaches <8%; b) Milling the dried plant material and passing through mesh size 60, for extraction through pulverization, to obtain a mixture; c) Boiling the mixture in 5-10 fold amount of water 3-6 times for 30 minutes each followed by fermenting the mixture to 24-36 hours at 30 to 350C; d) Incubating the mixture obtained in step (c) with microbes Lactobacillus spp. having CFU in the range of 2 × 104to 3 × 106and transferring the mixture into a vessel followed by fermenting the mixture to 24-36 hours at 30 to 350C, to obtain a solution; 1B5156-00007 e) Filtering the solution to remove fine particles followed by heating for 30 min at 60-90°C to obtain a solution; f) Concentration the solution obtained in the above step under low temperature to extract until its relative density reaches from 1.1 to 1.4; g) Mixing the concentrated extract in a blender for 15-30 min with lemon and pomegranate Juice or a mixture thereof present in an amount of 0.5 to 2.5% based on the total weight of plant material.

3. A herbal nano composition improving Vascular Activity and Cardiac Health comprising: a) Composition as claimed in claim 1; b) Excipients selected from D-α-Tocopherol polyethylene glycol 1000 succinate present in an amount ranging from 0.01 - 0.02%, Copolyvidone present in an amount ranging from 0.20 - 2.00%, Propylene glycol monocaprylate present in an amount ranging from 0.50 - 2.00%, polyethylene glycol present in an amount ranging from 0.75 - 2.00%, lecithin present in an amount ranging from 0.10 - 5.00% and Polyoxyethylene lauryl ether present in an amount ranging from 0.01 - 0.10%.

4. A process of preparing the herbal nano composition as claimed in claim 3, comprises the steps of: a) Mixing the composition as claimed in claim 1 along with the excipients selected from D-α-Tocopherol polyethylene glycol 1000 succinate present in an amount ranging from 0.01 - 0.02%, Copolyvidone present in an amount ranging from 0.20 - 2.00%, Propylene glycol monocaprylate present in an amount ranging from 0.50 - 2.00%, polyethylene glycol present in an amount ranging from 0.75 - 2.00%, lecithin present in an amount ranging from 0.10 - 5.00% and Polyoxyethylene lauryl ether present in an amount ranging from 0.01 - 0.10%; b) Sonicating the admixture for 25 to 30 minutes in a probe sonicator at an amplitude ranging between 20 to 45% and temperature between 25 to 45°C, to obtain a nano- sized admixture; c) Homogenising the nano-sized admixture for 5-10 minutes at 10,000 to 15,000 RPM, to obtain a synergistic nano herbal composition.

5. The process as claimed in claim 1, wherein the plant material comprises Plant bark and seeds. 2

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