Polyherbal composition for the management of hypertension and related disorders and method of preparation thereof

Abstract

The present invention relates to the field of pharmacology and herbal medicine. More particularly relates to the development of a novel polyherbal composition for the management of hypertension and related stress.

Description

[0001] B5156-00009

[0002] POLYHERBAL COMPOSITION FOR THE MANAGEMENT OF HYPERTENSION AND RELATED DISORDERS AND METHOD OF PREPARATION THEREOF

[0003] FIELD OF THE INVENTION:

[0004] The present invention relates to the field of pharmacology and herbal medicine. More particularly relates to the development of a novel polyherbal composition for the management of hypertension and related stress.

[0005] BACKGROUND OF THE INVENTION:

[0006] 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 specifically or implicitly referenced is prior art.

[0007] Hypertension, or high blood pressure, affects millions globally, acting as a significant risk factor for cardiovascular diseases, stroke, and kidney failure. Despite the availability of various antihypertensive drugs, long-term usage often results in adverse effects such as dizziness, fatigue, electrolyte imbalances, and reduced patient compliance due to high costs and side effects. These challenges necessitate the development of safer, affordable, and efficacious alternatives.

[0008] Herbal formulations are gaining traction due to their perceived safety, affordability, and minimal side effects. However, the lack of standardization, rigorous scientific validation, and comprehensive understanding of interactions has hindered the development of reliable and effective herbal-based treatments for hypertension. Current pharmaceutical treatments, while effective, fail to address this gap, especially for patients seeking natural alternatives.

[0009] Although traditional pharmacological treatments yield positive results, they often come with undesired side effects, such as low blood pressure, dizziness, and potential organ damage, which may not be suitable for all individuals.

[0010] Thus, there exists an unmet need for a scientifically validated, standardized, and efficacious herbal formulation to manage hypertension and its associated stress effectively.

[0011] Particularly, there is a need of an efficient polyherbal composition for the management of hypertension and related stress.B5156-00009

[0012] OF THE INVENTION:

[0013] The primary object of the present invention is to overcome the drawbacks associated with prior art by providing a novel polyherbal composition for the management of hypertension and related stress.

[0014] Another objective of present invention is to provide an innovative polyherbal solution for hypertension, focusing on establishing a therapeutic alternative that is safe, effective, and affordable, while overcoming the shortcomings associated with prevailing antihypertensive medications. Hypertension remains a critical global health issue, significantly impacting cardiovascular health and increasing the risk of associated diseases.

[0015] Another objective of present invention is to provide a polyherbal remedy, which offers promising avenues as either complementary or alternative strategies for managing hypertension.

[0016] Another objective of present invention is to provide a polyherbal composition comprising natural ligands to target multiple mechanistic pathways like Aldosterone, ACE, and Endothelin-1, either individually or in combination, with enhanced efficacy.

[0017] An objective of present invention is to provide a process of the preparation of novel polyherbal composition for the management of hypertension and related stress.

[0018] SUMMARY OF THE INVENTION:

[0019] This summary is provided to introduce a selection of concepts, in a simplified format, that are further described in the detailed description of the invention. This summary is neither intended to identify key or essential inventive concepts of the invention and nor is it intended for determining the scope of the invention.

[0020] In an aspect, the invention provides a polyherbal composition for the management of hypertension and related stress comprising the components:

[0021] a) Boerhavia diffusa present in an amount ranging from 1.0-3.5% by weight; b) Withania somnifera present in an amount ranging from 2.0-5.5% by weight; c) Tribulus terrestris present in an amount ranging from 1.5-4.5% by weight; d) Allium sativum present in an amount ranging from 1.5-5.5% by weight;

[0022] e) Terminalia arjuna present in an amount ranging from 2.1-5.6% by weight;B5156-00009

[0023] f) Moringa oleifera present in an amount ranging from 0.5-2.5% by weight; g) Butea parviflora present in an amount ranging from 1.25-4.5% by weight; h) Acalypha fruticosa present in an amount ranging from 1.25-4.5% by weight. A process of preparing the polyherbal composition for the management of hypertension and related stress, comprising the steps of:

[0024] (a) Mixing the composition obtained as in claim 1 with one or more excipients selected from poloxamer 188, hydroxypropyl methylcellulose (HPMC), medium-chain triglycerides (MCT oil), PEG 400, phosphatidylcholine, and sorbitan monooleate; (b) Sonicating the mixture for 20 to 30 minutes at 25-50% amplitude and 30-50°C to form a nano-sized mixture;

[0025] (c) Homogenizing the nano-sized mixture for 5 to 10 minutes at 12,000 to 16,000 RPM to obtain a stable, synergistic nano herbal composition.

[0026] The Invention also provides a method of obtaining extract of the herbs for the composition as described above, which comprises the steps of:

[0027] a) Whole plant, dried leaves, or roots were mixed with 30-60% alcohol at a ratio of 1:30 (g / mL);

[0028] b) mixture was agitated at 50-500 rpm at room temperature, followed by sonication for 40-80 minutes at 25-45°C;

[0029] c) After 2 to 3 day incubation period with intermittent shaking, activated charcoal was added to remove impurities;

[0030] d) The mixture was stirred for 30-60 minutes, filtered, and the solvent was evaporated under reduced pressure (60-150 mBar) at 40-70°C.

[0031] In another aspect, the extract of the bark and woody materials is obtained from the steps of:

[0032] a) dried plant material was pulverized in 40-100 mesh, followed by extraction with a hydro-alcoholic solution comprising 20-60% alcohol;

[0033] b) filtration and solvent evaporation under similar reduced pressure and temperature conditions.B5156-00009

[0034] DETAILED DESCRIPTION OF DRAWINGS:

[0035] To further clarify the advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which are illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting in their scope. The invention will be described and explained with additional specificity and detail with the accompanying drawings in which:

[0036] Figure 1 illustrates results of DPPH inhibition of various compositions

[0037] Figure 2 illustrates results of synergy index of various compositions

[0038] Figure 3 illustrates size of the nanoparticles of final composition (i.e. Composition 1224e) Figure 4 illustrates In vitro permeability of the present invention (i.e. Composition 1224e) as compared to the conventional composition.

[0039] Figure 5 illustrates comparison of systolic pressure of Composition 1224e versus a Normal control, Disease control and Chemical control.

[0040] Figure 6 illustrates comparison of diastolic pressure of Composition 1224e versus a Normal control, Disease control and Chemical control.

[0041] Figure 7 illustrates comparison of mean arterial pressure of composition 1224e versus a Normal control, Disease control and Chemical control.

[0042] Figure 8 illustrates comparison of cholesterol (mg / dl) of composition 1224e versus a Normal control, Disease control and Chemical control.

[0043] Figure 9 illustrates comparison of Triglycerides (mg / dl) of composition 1224e versus a Normal control, Disease control and Chemical control.

[0044] Figure 10 illustrates comparison of LDL (mg / dl) of composition 1224e versus a Normal control, Disease control and Chemical control

[0045] Figure 11 illustrates comparison of systolic pressure of composition 1224e versus a Normal control, Disease control and Chemical control

[0046] Figure 12 illustrates comparison of diastolic pressure of composition 1224e versus a Normal control, Disease control and Chemical controlB5156-00009

[0047] Figure 13 illustrates comparison of mean arterial pressure of composition 1224e versus a Normal control, Disease control and Chemical control

[0048] Figure 14 illustrates comparison of cholesterol (mg / dl) of composition 1224e versus a Normal control, Disease control and Chemical control

[0049] Figure 15 illustrates comparison of HDL (mg / dl) of composition 1224e versus a Normal control, Disease control and Chemical control

[0050] Figure 16 illustrates comparison of Triglycerides (mg / dl) of composition 1224e versus a Normal control, Disease control and Chemical control

[0051] Figure 17 illustrates comparison of LDL (mg / dl) of composition 1224e versus a Normal control, Disease control and Chemical control

[0052] DETAILED DESCRIPTION:

[0053] 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.

[0054] 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.

[0055] The term “Composition” refers to preparation of which are in such form as to permit the biological activity of the active ingredients to be unequivocally effective. The term “Formulation” or “Composition” are similar and can be used here inter-changeably

[0056] The present invention relates to the development of a novel polyherbal composition for the management of hypertension and related stress. This invention integrates molecular docking studies, in vitro assays, and in vivo efficacy evaluations to identify and validate a synergistic formulation using natural extracts. The invention also encompasses nano formulations for enhanced bioavailability and therapeutic efficacy.B5156-00009

[0057] In a preferred embodiment, the invention provides a polyherbal composition for the management of hypertension and related stress which comprises following components:

[0058] a) Boerhavia diffusa present in an amount ranging from 1.0-3.5% by weight; b) Withania somnifera present in an amount ranging from 2.0-5.5% by weight; c) Tribulus terrestris present in an amount ranging from 1.5-4.5% by weight; d) Allium sativum present in an amount ranging from 1.5-5.5% by weight; e) Terminalia arjuna present in an amount ranging from 2.1-5.6% by weight; f) Moringa oleifera present in an amount ranging from 0.5-2.5% by weight; g) Butea parviflora present in an amount ranging from 1.25-4.5% by weight; h) Acalypha fruticose present in an amount ranging from 1.25-4.5% by weight. The experiments were performed to determine the synergistic activity of the claimed composition. The results show that composition 1224e has the lowest IC50 of 17.53 mg / mL value for antioxidant activity as compared to the IC50 of the individual herb / s as mentioned in TABLE 1.

[0059] In an embodiment, the novel polyherbal nano-formulation, Formulation / Composition 1224e, was developed by combining various extracts based on their synergistic antioxidant potential, which was further screened using inhibition potential against Angiotensin Convertase Enzyme and lipase. The efficacy of this composition was then validated using well-established animal models for hypertension, such as DOCA Salt induced hypertension and L-NAME induced Hypertension. The results demonstrated that the formulation / composition 1224e exhibited beneficial effects in both of these animal models, showing promising therapeutic potential for the management of hypertension. The formulation / composition 1224e is useful in conditions such as Management of stress, Cardiovascular disease, and metabolic disorders, including dyslipidemia

[0060] It also helps in up-regulating the levels of HDL and lowering of Cholesterol, LDL and Triglycerides metabolic diseases.

[0061] In accordance of the present invention, the polyherbal composition targets multiple mechanistic pathways for effective management for hypertension and related conditions. These pathways include, but are not limited to, ACE inhibition, Endothelin-1 antagonism, Nitric oxide modulation, Anti-oxidant activity, Beta-adrenergic receptor modulation, and lipase inhibition.B5156-00009

[0062] The polyherbal composition demonstrates its antihypertensive potential through these multiple mechanism, resulting in but not limited to reduced vasoconstriction, oxidative stress and vascular resistance. The composition is designed to exhibit redundancy, ensuring that if one mechanistic pathway is inhibited, alternative pathways are activated or become predominant thereby maintaining therapeutic efficacy of hypertension management.

[0063] Further pharmacological studies and clinical findings strongly justify the compositions multitarget approach, supported by drug-likeness and ADMET analyses, which confirm strong adherence to Lipinski's Rule of Five, optimal bioavailabity and minimal toxicity of the composition. Additionally molecular studies reveal superior binding affinities for hypertension-related targets such as aldosterone, neprilysin, ACE and endothelin- 1. This redundancy and interplay among multiple pathways ensure that the composition remains effective under varying physiological conditions, supported further by safe toxicity profile. In accordance with the present invention, the claimed polyherbal composition can be used as an adjuvant in combination with a chemical and / or non-chemical antihypertensive and diuretic agent, including but not limited to Captopril, Bostean, Enalapril and Spironolactone. In an embodiment, the said combination therapeutically targets the specific receptors intended, such as the Aldosterone (Spironolactone), and ACE receptors (Enalapril), thereby modulating key pathways involved in the regulation of hypertension. Although the primary action is directed towards the said receptors, the composition may also exert off-target effects by influencing additional receptors implicated in the pathophysiology of hypertension, thereby contributing to a broader, more comprehensive therapeutic effect. The synergistic effects of these combinations result in a more potent antihypertensive action, offering a comprehensive therapeutic strategy for regulation of hypertension.

[0064] In another embodiment, the invention provides a process of preparing the polyherbal composition for the management of hypertension and related stress. The process comprises following steps:

[0065] (a) Mixing the composition obtained as in claim 1 with one or more excipients selected from poloxamer 188, hydroxypropyl methylcellulose (HPMC), medium-chain triglycerides (MCT oil), PEG 400, phosphatidylcholine, and sorbitan monooleate; (b) Sonicating the mixture for 20 to 30 minutes at 25-50% amplitude and 30-50°C to form a nano-sized mixture;B5156-00009

[0066] (c) Homogenizing the nano-sized mixture for 5 to 10 minutes at 12,000 to 16,000 RPM to obtain a stable, synergistic nano herbal composition.

[0067] In another embodiment, the invention provides a method of obtaining extract of the herbs for the composition as described above, which comprises the steps of:

[0068] a) Whole plant, dried leaves, or roots were mixed with 30-60% alcohol at a ratio of 1:30 (g / mL);

[0069] b) mixture was agitated at 50-500 rpm at room temperature, followed by sonication for 40-80 minutes at 25-45°C;

[0070] c) After 2 to 3 day incubation period with intermittent shaking, activated charcoal was added to remove impurities;

[0071] d) The mixture was stirred for 30-60 minutes, filtered, and the solvent was evaporated under reduced pressure (60-150 mBar) at 40-70°C.

[0072] In another embodiment, the extract of the bark and woody materials is obtained from the steps of:

[0073] a) dried plant material was pulverized in 40-100 mesh, followed by extraction with a hydro-alcoholic solution comprising 20-60% alcohol;

[0074] b) filtration and solvent evaporation under similar reduced pressure and temperature conditions.

[0075] The invention is described with the help of various experiments as explained below:

[0076] METHOD OF PREPARATION:

[0077] Extraction process:

[0078] Method A: Whole plant, dried leaves, or roots were mixed with 30-60% alcohol at a ratio of 1:30 (g / mL). The mixture was agitated at 50-500 rpm at room temperature, followed by sonication for 40-80 minutes at 25-45°C. After a 2-3 day incubation period with intermittent shaking, activated charcoal was added to remove impurities. The mixture was stirred for 30-60 minutes, filtered, and the solvent was evaporated under reduced pressure (60-150 mBar) at 40-70°C.

[0079] Method B: For bark and woody materials, dried plant material was pulverized (40-100 mesh). Extraction was performed with a hydro -alcoholic solution (20-60% alcohol), followed by filtration and solvent evaporation under similar reduced pressure and temperature conditions.B5156-00009

[0080] Efficacy evaluation

[0081] In vitro

[0082] Respective extracts were screened for antioxidant potential using DPPH based screening assay.

[0083] Method: Free radical scavenging activity was assessed using the DPPH assay. A 200 pM DPPH solution in methanol was mixed with test compounds in varying concentrations. After 30 minutes, absorbance was measured at 517 nm.

[0084] Absorbance of control - Absorbance of test

[0085] % Scavenging = X 100 Absorbance of control

[0086] TABLE 1: IC50 Values for DPPH Scavenging Activity of Individual Herb Extracts

[0087]

[0088] Formulation Development

[0089] Dried extracts were blended in varying ratios with excipients, i.e., pharmacologically inactive components, such as but not limited to binders, dis-integrant, stabilizers, etc. which are generally recognized as safe, non-toxic, and acceptable for human pharmaceutical use. These auxiliary materials aid in the development of multiple dosage forms, including powders, tablets, capsules, and syrups. For example, the binders include, but are not limited to, cellulose, starch, gum acacia, and pectin; the dis-integrant include, but are not limited to, croscarmellose sodium and sodium starch glycolate; and the stabilizers include, but are not limited to, tocopherols, squalene, tartaric acid, and glycerin. These excipients play crucial roles in ensuring the stability, integrity, and effective delivery of the active ingredients in the polyherbal composition.

[0090] Synergistic activity was explored using combinations. Composition 1224e showed the lowest IC50 (17.53 mg / mL) value for antioxidant activity.B5156-00009

[0091] TABLE 2: Composition 1

[0092]

[0093] TABLE 3: Composition 2

[0094]

[0095] TABLE 4: Composition 1224e

[0096]

[0097] TABLE 5: DPPH Activity IC50 Values for Different Compositions

[0098]

[0099] FIGURE 1 shows composition 1224e exhibits synergism when compared to the %inhibition observed with composition 1 & 2 which indicates a greater inhibitory effect from the composition 1224e than from individual extracts alone.

[0100] The composition 1224e produces a synergistic enhancement in DPPH radical scavenging activity, as evidenced by the significant reduction in IC50 to 17.53 mg / mL. This suggests thatB5156-00009

[0101] the mixtures, when combined, provide greater antioxidant potential than anticipated, highlighting the importance of exploring synergistic effects in the development of novel antioxidant formulations for therapeutic or nutraceutical applications.

[0102] FIGURE 2 show that Composition 1224e exhibits the most potent antioxidant activity when compared to the individual samples and other compositions, indicating a synergistic effect. Further Screening of developed compositions was carried out using methods such as Lipase inhibition action and Angiotensin convertase enzyme inhibitory action.

[0103] Lipase Assay: Porcine pancreatic lipase was used, with para-nitrophenylbutyrate (PNPB) as the substrate. Test samples were incubated with enzyme solution and phosphate buffer (pH 7.2) at 37°C. After substrate addition, absorbance was measured at 400 nm.

[0104] TABLE 6: Lipase Inhibition IC50 Values for Different Compositions

[0105]

[0106] ACE inhibition Assay: Angiotensin-converting enzyme (ACE) activity was measured by monitoring the release of HA from the substrate HHL. The yellow color developed was quantified at 410 nm.

[0107] TABLE 7: ACE Inhibition IC50 Values for Different Compositions

[0108]

[0109] These results suggest that Composition 1224e is the most promising candidate for further development of formulation for the management of vascular stress NANOFORMULATION

[0110] The solubility and compatibility of Composition 1224e were evaluated using various excipients such as, including D Poloxamer 188, Hydroxypropyl methylcellulose (HPMC), Medium-chainB5156-00009

[0111] triglycerides (MCT oil), PEG 400, Phosphatidylcholine, and Sorbitan monooleate. The admixture was sonicated for 20 to 30 minutes at 25-50% amplitude and 30-50°C, resulting in a nano-sized mixture. It was then homogenized for 5 to 10 minutes at 12,000 to 16,000 RPM to form a stable, synergistic nano herbal composition.

[0112] Figure 3 shows the average nano particle size obtained was in the range of 215-546nm. COMPARATIVE IMPROVEMENT IN PERMEABILITY:

[0113] Permeability studies were performed using Everted Gut Sac method. Ileum was cut into 10-12 cm long pieces and gently everted. Gut Sac were prepared and filled with 5 ml of ringer solution. Prepared Gut Sacs were placed in beaker containing 50 ml of 1000 ppm Formula 1224e under constant agitation and aeration at 35-37°C for 90 min.

[0114] After 90 minutes gut sacs were removed and outer surface was washed thoroughly. Sacs were opened and ringer solution was collected in pre-labelled tubes. HPTLC based Densitometry quantification was carried out.

[0115] FIGURE 4 shows in vitro permeability of the formulation as compared to a conventional composition.

[0116] QUALITY CONTROL PARAMETERS OF DEVELOPED FORMULATION

[0117] Quantification of secondary compounds was conducted for the Composition 1224e to ensure consistency and potency across different batches, thus aiding in achieving therapeutic goals.

[0118] TABLE 8: Quality Control Parameters of Composition 1224e

[0119]

[0120] MULTIPLE MECHANISTIC PATHWAYS AND DRUG-LIKENESS PROFILING: The polyherbal formulation targets multiple pathways involved in hypertension:

[0121] ACE Inhibition: Reduces angiotensin II production, lowering vasoconstriction and blood pressure.

[0122] Endothelin-1 Antagonism: Mitigates vasoconstriction and vascular resistance.

[0123] Antioxidant Activity: Combats oxidative stress, a key contributor to endothelial dysfunction.B5156-00009

[0124] Nitric Oxide Modulation: Enhances vasodilation by upregulating nitric oxide synthase (NOS3) activity.

[0125] Beta-Adrenergic Receptor Modulation: Reduces heart rate and myocardial contractility.

[0126] The drug-likeness, efficacy, physicochemical, and pharmacokinetic properties of the identified phytochemicals were analyzed using computational tools like SwissADME, pkCSM, and SwissDock. Key evaluations were based on Lipinski's Rule of Five (RO5) and ADMET criteria (Absorption, Distribution, Metabolism, Excretion, and Toxicity).

[0127] Most compounds adhered to RO5, demonstrating favorable physicochemical properties such as an acceptable range of molecular weight (MW), hydrogen bond acceptors (HBA), hydrogen bond donors (HBD), and logP (lipophilicity). For instance:

[0128] Caffeic acid (MW: 180.16, LogP: 1.15, HBA: 4, HBD: 3, Bioavailability Score: 0.56) met all parameters without any violations.

[0129] Kaempferol (MW: 286.23, LogP: 2.28, HBA: 6, HBD: 4, Bioavailability Score: 0.55) adhered to RO5 with no violations.

[0130] Gallic acid (MW: 170.12, LogP: 0.70, HBA: 5, HBD: 4, Bioavailability Score: 0.56) also fulfilled all the criteria.

[0131] Eupalitin (MW: 330.29, LogP: 2.82, HBA: 7, HBD: 3, Bioavailability Score: 0.55) demonstrated excellent compliance with the rules.

[0132] These examples highlight the promising drug-like properties of these compounds, with no violations detected in key parameters, thereby supporting their potential as therapeutic candidates. This systematic profiling strengthens the case for their further pharmacological investigation

[0133] TABLE 9: Drug Likeness Properties and Bioavailability Score of Selected Ligands

[0134]

[0135] B5156-00009

[0136]

[0137] In terms of safety, the phytochemicals were evaluated for their LD50, hepatotoxicity, carcinogenicity, and mutagenicity. The data highlights compounds with favourable safety profiles, such as:

[0138] 5 Chlorogenic acid (LD50: 5000 mg / kg) with no hepatotoxicity, carcinogenicity, or mutagenicity.

[0139] Kaempferol (LD50: 3919 mg / kg), Niazirin (LD50: 3750 mg / kg), and Niaziridin (LD50: 4000 mg / kg) with no toxicity, carcinogenicity, or mutagenicity.

[0140] Allicin (LD50: 874 mg / kg), Quercetin (LD50: 159 mg / kg), and Oleanolic acid (LD50: 2000 10 mg / kg) also show no mutagenicity, with negligible demonstrating carcinogenic or hepatotoxic activity.

[0141] Some compounds did show concerns in specific areas, such as:

[0142] Quercetin (LD50: 159 mg / kg) with both carcinogenic and mutagenic activity.

[0143] Oleanolic acid (LD50: 2000 mg / kg) demonstrated both hepatotoxic and carcinogenic activity. 15 TABLE 10: Toxicity prediction of selected phytochemicals compounds

[0144]

[0145] B5156-00009

[0146]

[0147] The docking study involves evaluating the binding affinity of various ligands against key target proteins, namely Aldosterone, Neprilysin, ACE (Angiotensin-Converting Enzyme), Endothelin-1, and SESN2. Binding affinities are measured in kcal / mol, with lower (more 5 negative) values indicating stronger binding. The amino acid interactions highlight the key residues in the protein that interact with each ligand.

[0148] TABLE 11: Docking results of selected compounds against Aldosterone

[0149]

[0150] B5156-00009

[0151]

[0152] Arjugenin, Sarsasapogenin, and Arjunic acid exhibited the strongest binding affinities (-10.59, -10.22, and -9.88 kcal / mol, respectively).

[0153] These ligands form hydrogen bonds and hydrophobic interactions with residues such as PHE D:381, CYS D:450, and GLU D:310, disrupting aldosterone's activityB5156-00009

[0154] TABLE 12: Docking results of selected compounds against Neprilysin

[0155]

[0156] B5156-00009

[0157] Chlorogenic acid demonstrated binding affinity of -6.97 kcal / mol, with robust interactions involving HIS A:711, ARG A:717, and GLU A:584.

[0158] TABLE 13: Docking results of selected compounds against ACE

[0159]

[0160] Eupalitin and Niazirin (-7.2 kcal / mol) showed the strongest inhibition, interacting with residues such as GLU A:208 and LYS A:562.

[0161]

[0162] B5156-00009

[0163]

[0164] Arjugenin (-5.95 kcal / mol) and Sarsasapogenin (-5.83 kcal / mol) interact with residues including LEU A:6, MET A:7, and HIS A: 16, which are critical for Endothelin-l's functionality.

[0165] TABLE 15: Docking results of selected compounds against SESN2

[0166]

[0167] Niaziridin (-5.94 kcal / mol) binds effectively to GLU A:135 and HIS A:132, offering potential for oxidative stress regulation.

[0168] The combination of ligand with Spironolactone and Enalapril results in significantly improved binding affinities and inhibition constants, as shown below:

[0169] TABLE 16: Binding Affinities and Inhibition Constants for Ligands and Their Combinations with Spironolactone targeting Aldosterone.

[0170]

[0171] B5156-00009

[0172]

[0173] TABLE 17: Binding Affinities and Inhibition Constants for Ligands and Their Combinations with Enalapril targeting ACE

[0174]

[0175] IN VIVO EFFICACY STUDIES STUDY 1 (DOCA induced Hypertension)

[0176] The DOCA salt model is used to induce secondary hypertension in rats by injection of DOCA salt and providing 1% saline drinking water, activating the renin-angiotensin-aldosterone system (RAAS) and increasing sodium retention. The study includes four experimental groups: Normal Control, which receives standard care and serves as a baseline; Disease Control, where hypertension is induced and no treatment; Chemical Control, where hypertension induced and receives a standard antihypertensive drug to evaluate its effect on blood pressure; and Composition 1224e, which tests the efficacy test formulation in mitigating hypertension. Blood pressure (systolic, diastolic, and mean arterial pressure) was measured on Day 0 and Day 42 to assess the progression of hypertension and the efficacy of treatments.

[0177] The disease control group experienced severe and uncontrolled increases in systolic, diastolic, and mean arterial pressures over 42 days, indicating significant hypertension progression without treatment. The chemical control group demonstrated excellent efficacy in maintainingB5156-00009

[0178] stable blood pressure, with minimal increases in all parameters, making it the most effective treatment. The composition 1224e treatment group showed efficacy, which was significantly better at controlling hypertension compared to disease group and there both chemical control and test formulations (composition 1224e) were equally effective in reducing the blood pressure when compared to disease group.

[0179] STUDY 2 (L-NAME induced Hypertension)

[0180] The L-NAME (Nco-Nitro-L-arginine methyl ester) induced hypertension model is used to simulate hypertension by inhibiting nitric oxide synthase, leading to reduced nitric oxide production, vasoconstriction, and increased blood pressure. In this study, rats are divided into four groups: Normal Control, which serves as the baseline with no treatment; Disease Control, where hypertension is induced by L-NAME treatment; Chemical Control, which receives the a standard antihypertensive drug to assess its effect on lowering blood pressure; and Composition 1224e, a novel treatment aimed at evaluating its potential antihypertensive effects. Blood pressure (systolic, diastolic, and mean arterial pressure) is measured at Day 28 to assess the efficacy of each treatment.

[0181] The Disease Control group exhibits significantly elevated cholesterol, triglycerides, and LDL, alongside reduced HDL. The Chemical control demonstrates an ability to bring these lipid markers closer to normal levels when regularly administered for 4 weeks. The composition 1224e treatment group showed efficacy, which was significantly equal at controlling hypertension compared to disease group.

Claims

B5156-00009We Claim:

1. A polyherbal composition for the management of hypertension and related disorders comprising the components:a) Boerhavia diffusa present in an amount ranging from 1.0-3.5% by weight; b) Withania somnifera present in an amount ranging from 2.0-5.5% by weight; c) Tribulus terrestris present in an amount ranging from 1.5-4.5% by weight; d) Allium sativum present in an amount ranging from 1.5-5.5% by weight; e) Terminalia arjuna present in an amount ranging from 2.1-5.6% by weight; f) Moringa oleifera present in an amount ranging from 0.5-2.5% by weight; g) Butea parviflora present in an amount ranging from 1.25-4.5% by weight; h) Acalypha fruticose present in an amount ranging from 1.25-4.5% by weight.

2. The composition as claimed in claim 1 , wherein the said composition is in dosage forms including, but not limited to, powders, tablets, capsules, syrups, and nano-formulations.

3. The composition as claimed in claim 1, wherein the said composition synergistically targets multiple mechanistic pathways.

4. The composition as claimed in claim 1, wherein the said composition comprises plurality of auxiliary materials selected from binders, dis-integrant and stabilizers.

5. The composition as claimed in claim 1, wherein the said composition is useful as an adjuvant in combination with one or more known chemical and / or non-chemical antihypertensive agents.

6. The composition as claimed in claim 1, wherein the disorders comprises hypertension, stress, cardiovascular diseases, and metabolic disorders, including dyslipidemia.

7. A process of preparing the polyherbal composition for the management of hypertension and related stress, comprising the steps of:a) Mixing the composition obtained in claim 1 with one or more excipients selected from poloxamer 188, hydroxypropyl methylcellulose, medium-chain triglycerides, PEG 400, phosphatidylcholine, and sorbitan monooleate;B5156-00009b) Sonicating the mixture for 20 to 30 minutes at 25-50% amplitude and 30-50°C to form a nano-sized mixture;c) Homogenizing the nano-sized mixture for 5 to 10 minutes at 12,000 to 16,000 RPM to obtain a stable, synergistic nano herbal composition.

8. A method of obtaining extract of the herbs for the composition as claimed in claim 1, comprising the steps of:a) Mixing the whole plant, dried leaves, or roots with 30-60% alcohol at a ratio of 1:30 (g / mL);b) Agitating of mixture at 50-500 rpm at room temperature, followed by sonication for 40-80 minutes at 25-45°C;c) Incubating for the period of 2 to 3 days with intermittent shaking, and adding activated charcoal to remove impurities;d) Stirring the mixture for 30-60 minutes, filtration, and the evaporation of solvent under reduced pressure (60-150 mBar) at 40-70°C.

9. A method of obtaining extract of the herbs for the composition as claimed in claim 1, comprising the steps of:a) Pulverization of bark and woody materials, dried plant material for 40-100 mesh, followed by extraction with a hydro -alcoholic solution comprising 20- 60% alcohol;b) Filtration and solvent evaporation under reduced pressure and temperature conditions as claimed in Claim 8.

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