Polyherbal compounds for managing hypertension and related disorders

Abstract

The present invention relates to a polyherbal compound based formulation for managing hypertension and related disorders, comprising Dioscin, Withanolide A, Withaferin A, Withanolide D, P-Sitosterol, Diosgenin, Arjunolic Acid, Caffeic Acid, and Kaempferol. The compounds targets multiple mechanistic pathways in hypertension, including the Renin- Angiotensin-Aldosterone System (RAAS), Sympathetic Nervous System, Vascular Resistance, Nitric Oxide, and Stress-Responsive pathways. It offers a comprehensive and synergistic approach for improving vascular health, reducing blood pressure, regulating the levels of HDL and lowering of Cholesterol, LDL and Triglycerides. The compound / s is formulated for enhanced bioavailability and efficacy, and can be used as an adjuvant with antihypertensive agents. The invention also includes methods for extracting, purifying, and standardizing the active ingredients, ensuring consistent quality. The compounds is suitable for managing hypertension, cardiovascular diseases, metabolic disorders, and stress. It can be administered in various dosage forms such as tablets, capsules, syrups, and powders.

Description

[0001] B5156-00012

[0002] POLYHERBAL COMPOUNDS FOR MANAGING HYPERTENSION AND RELATED DISORDERS FIELD OF INVENTION:

[0003] The present invention relates to field of pharmacology and herbal medicine. More particularly relates to the development of a novel polyherbal compound based formulation for the management of hypertension and related stress. The invention also provides nano-formulation for enhanced bioavailability and therapeutic efficacy.

[0004] BACKGROUND OF THE INVENTION:

[0005] 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.

[0006] Hypertension, or high blood pressure, is a common cardiovascular condition impacting millions globally. It is a major risk factor for heart disease, stroke, and kidney failure. While various pharmaceutical anti-hypertensive medications exist, long-term use often leads to adverse effects like dizziness, fatigue, and electrolyte imbalances. Additionally, patient non-compliance due to high medication costs and drug-related impediments further complicates hypertension management.

[0007] There is growing interest in natural and alternative therapies, particularly herbal formulations, for managing high blood pressure. This is due to their perceived safety, affordability, and lower side effects. However, despite the potential benefits of herbs with blood pressure lowering properties, there is a significant lack of scientifically validated, standardized, and clinically effective herbal formulations for hypertension. The absence of rigorous research, standardization, and comprehensive understanding of how herbs interact in these formulations poses a challenge in developing reliable and effective herbal-based treatments for high blood pressure.

[0008] Hypertension remains a critical global health issue, significantly impacting cardiovascular health and increasing the risk of associated diseases. 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.B5156-00012

[0009] Antihypertensive drugs are extensively employed to manage the high blood pressure. These medications can be categorized into various pharmacological classes, each exhibiting unique mechanisms of action and associated limitations.

[0010] Angiotensin-converting enzyme inhibitors, such as lisinopril and enalapril, function by impeding the conversion of angiotensin I to angiotensin II, thereby reducing blood pressure. However, these medications commonly elicit side effects including a persistent dry cough, hyperkalemia, and kidney dysfunction. Additionally, they are contraindicated in pregnant women and individuals with certain renal conditions.

[0011] Angiotensin II receptor blockers, including losartan and valsartan, also target the angiotensin system, but they do so by preventing angiotensin II from constricting blood vessels. While ARBs avoid the dry cough associated with ACE inhibitors, they can still cause hyperkalemia and dizziness, and may exhibit reduced efficacy in certain populations.

[0012] Calcium channel inhibitors, such as amlodipine and verapamil, promote vasodilation by restricting calcium influx into cardiac and vascular cells. However, these agents commonly induce side effects including ankle edema, dizziness, and, in certain instances, cardiovascular complications.

[0013] Beta-adrenergic antagonists, like metoprolol and atenolol, lower blood pressure by decreasing heart rate and myocardial contractility, yet are associated with fatigue, bradycardia, and respiratory distress in asthmatic individuals.

[0014] Diuretic medications, exemplified by hydrochlorothiazide and furosemide, facilitate the excretion of excess sodium and water, thereby reducing blood volume. Nonetheless, their utilization may precipitate electrolyte disturbances, dehydration, polyuria, and in some cases, elevated blood glucose and cholesterol levels.

[0015] Alpha-adrenergic receptor blockers like doxazosin and prazosin mediate vasodilation, but their utility is constrained by side effects including dizziness, fatigue, and orthostatic hypotension. Similarly, direct renin inhibitors such as aliskiren can lower blood pressure by directly inhibiting renin, but their use is limited by gastrointestinal and electrolyte disturbances, as well as potential renal complications.

[0016] Vasodilators like hydralazine and minoxidil work by relaxing vascular smooth muscle, yet often induce tachycardia, headaches, and fluid retention.B5156-00012

[0017] Sodium nitroprusside can cause cyanide toxicity with prolonged use or high doses, leading to symptoms like confusion, weakness, and metabolic acidosis. It may also result in hypotension, reflex tachycardia, and occasional nausea or vomiting.

[0018] Nitroglycerin commonly causes headaches due to cerebral vasodilation, along with flushing, hypotension, and reflex tachycardia. Prolonged use of nitroglycerin can lead to tolerance, reducing its efficacy.

[0019] Centrally acting antihypertensive agents, exemplified by clonidine and methyldopa, reduce blood pressure by dampening sympathetic nervous system activity, but their sedative properties and the risk of rebound hypertension upon abrupt discontinuation constrain their clinical application.

[0020] Therefore, the existing knowledge lacks a solution that addresses the above mentioned problems / issues and hence there is a need of an efficient treatment such as poly herbal compounds for the management of hypertension and related stress.

[0021] OBJECT(S) OF THE PRESENT INVENTION:

[0022] The primary objective of the present invention is to overcome the drawback associated with prior art.

[0023] An objective of the invention is to provide an innovative polyherbal compound based formulation for hypertension and vascular stress, focusing on establishing a therapeutic alternative that is safe, effective, and affordable, while overcoming the shortcomings associated with prevailing antihypertensive medications.

[0024] Another objective of the invention is to provide a novel therapeutic polyherbal compound based nano-formulation for better permeability, enhance bioavailability and efficacy that acts via multiple mechanistic pathways.

[0025] Another objective of the invention is to provide a polyherbal compound based formulation that helps in up- regulating the levels of HDL and lowering levels of Cholesterol, LDL and Triglycerides.

[0026] Another objective of the invention is to provide a polyherbal compound based formulation for the treatment of metabolic diseases and conditions like atherosclerosis and pulmonary arterial hypertension.B5156-00012

[0027] SUMMARY OF THE INVENTION:

[0028] An object of the invention is to provide a polyherbal compound based formulation for the management of hypertension and other related disorders comprising the following constituents of phyto compounds:

[0029] a) Dioscin present in an amount ranging from 0.16 to 0.70%;

[0030] b) Withanolide A present in an amount ranging from 1.20 to 6.00%;

[0031] c) Withaferin A present in an amount ranging from 0.5 to 3.6%;

[0032] d) Withanolide D present in an amount ranging from 0.51 to 2.40%;

[0033] e) P-Sitosterol present in an amount ranging from 1.2 to 3.70%;

[0034] f) Diosgenin present in an amount ranging from 1.30 to 6.10%;

[0035] g) Arjunolic Acid present in an amount ranging from 0.15 to 1.20%;

[0036] h) Caffeic Acid present in an amount ranging from 0.02 to 1.40%;

[0037] i) Kaempferol present in an amount ranging from 0.20 to 1.30%.

[0038] BRIEF DESCRIPTION OF DRAWINGS:

[0039] The accompanying drawings, which are incorporated in and constitute a part of this disclosure, illustrate exemplary embodiments and, together with the description, explain the disclosed principles. The reference numbers are used throughout the figures to describe the features and components. Some embodiments of system and / or methods in accordance with embodiments of the present subject matter are now described, by way of example only, and regarding the accompanying figures, in which:

[0040] Figure 1: illustrates the method for extraction and standardization.

[0041] Figure 2: Illustrate the column chromatography method for PEc.

[0042] Figure 3: illustrates the method of preparation of PEc based solid dosage form.

[0043] Figure 4: illustrates the preparation of PEc based liquid dosage form.

[0044] Figure 5a: illustrates impact on systolic pressure by the compounds of present invention as compared to a control.

[0045] Figure 5b: illustrates impact on Diastolic pressure by the compounds of present invention as compared to a control.B5156-00012

[0046] Figure 5c: illustrates impact on mean arterial pressure by the compounds of present invention as compared to a control.

[0047] Figure 5d: illustrates impact on Cholesterol (mg / dl) by the compounds of present invention as compared to a control.

[0048] Figure 5e: illustrates impact on Triglycerides (mg / dl) by the compounds of present invention as compared to a control.

[0049] Figure 5f: illustrates impact on HDL (mg / dl) by the compounds of present invention as compared to a control.

[0050] Figure 5g: illustrates impact on LDL (mg / dl) by the compounds of present invention as compared to a control.

[0051] Figure 6a: illustrates impact on systolic pressure by the compounds of present invention as compared to a control.

[0052] Figure 6b: illustrates impact on Diastolic pressure by the compounds of present invention as compared to a control.

[0053] Figure 6c: illustrates impact on mean arterial pressure by the compounds of present invention as compared to a control.

[0054] Figure 6d: illustrates impact on Cholesterol (mg / dl) by the compounds of present invention as compared to a control.

[0055] Figure 6e: illustrates impact on HDL (mg / dl) by the compounds of present invention as compared to a control.

[0056] Figure 6f: illustrates impact on Triglycerides (mg / dl) by the compounds of present invention as compared to a control.

[0057] Figure 6g: illustrates impact on LDL (mg / dl) by the compounds of present invention as compared to a control.

[0058] Figure 7: illustrates the chromatographic separation of bands of compounds present in the formulation along with bands of standard marker compounds for Gallic acid and Ferulic acid.B5156-00012

[0059] DETAILED DESCRIPTION OF THE INVENTION:

[0060] In the present document, the word "exemplary" is used herein to mean "serving as an example, instance, or illustration." Any embodiment or implementation of the present subject matter described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments.

[0061] While the disclosure is susceptible to various modifications and alternative forms, specific embodiment thereof has been shown by way of example, in the drawings and will be described in detail below. It should be understood, however, that it is not intended to limit the disclosure to the specific forms disclosed, but on the contrary, the disclosure is to cover all modifications, equivalents, and alternative falling within the spirit and the scope of the disclosure.

[0062] 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 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.

[0063] The present invention provides an innovative polyherbal compound based formulation for hypertension, focusing on establishing a therapeutic alternative that is safe, effective, and affordable, while overcoming the shortcomings associated with prevailing antihypertensive medications.

[0064] The invention provides a scientifically supported and regulatory-compliant polyherbal compound based formulation that provides sustainable and comprehensive management of hypertension, enhances patient adherence, and meets the increasing interest in natural and integrative healthcare options.

[0065] This invention provides a novel therapeutic approach utilizing natural ligands to target multiple mechanistic pathways associated with hypertension and related disorders. These includes, but are not limited to, Renin-Angiotensin-Aldosterone System (RAAS), targeting ACE (angiotensin-converting enzyme), Angiotensin II receptors and Aldosterone; Sympathetic Nervous System pathways, including P-Adrenergic Receptors (P-AR) and Calcium Channel, Voltage-Dependent, Beta 1 Subunit (CACNB1); Vascular Resistance Pathways, includingB5156-00012

[0066] Endothelin-1 and Soluble Guanylate Cyclase (sGC); Nitric Oxide Pathways, involving Nitric Oxide Synthase 3 (NOS3) and Asymmetric Dimethylarginine (ADMA); Neprilysin and Stress-Responsive Pathways, including Sestrin-2 (SESN2), to improve vascular health and reduce blood pressure.

[0067] The Invention provides a novel polyherbal compound based formulation (also referred as C 1224c hereinafter) formulated using in vitro method to target ACE inhibition, Endothalin-1 regulation, stress-responsive protein Sestrin-2 (SESN2), protease (Neprilysin). The efficacy of the formulation is also validated using well-established animal models for hypertension, such as DOCA Salt induced hypertension and L-NAME induced Hypertension. The results demonstrated that the compounds C 1224c exhibited beneficial effects in both of these animal models, showing promising therapeutic potential for the management of hypertension. The compounds C 1224c is also useful in following conditions such as Management of stress and cardiovascular disease

[0068] It also helps in up regulating the levels of HDL and lowering of Cholesterol, LDL and Triglycerides.

[0069] It can be useful in metabolic diseases and conditions like atherosclerosis and pulmonary arterial hypertension.

[0070] In an embodiment, the compounds comprises:

[0071] a) Dioscin in an amount of 0.16 to 0.70% by weight;

[0072] b) Withanolide A in an amount of 1.20 to 6.00% by weight;

[0073] c) Withaferin A in an amount of 0.5 to 3.6% by weight;

[0074] d) Withanolide D in an amount of 0.51 to 2.40% by weight;

[0075] e) P-Sitosterol in an amount of 1.2 to 3.70% by weight;

[0076] f) Diosgenin in an amount of 1.30 to 6.10% by weight;

[0077] g) Arjunolic Acid in an amount of 0.15 to 1.20% by weight;

[0078] h) Caffeic Acid in an amount of 0.02 to 1.40% by weight; and

[0079] i) Kaempferol in an amount of 0.20 to 1.30% by weight.

[0080] The compounds are effective in regulating stress and managing hypertension.B5156-00012

[0081] In an embodiment, the compounds may comprise herbal component with function similar to Dioscin present in an amount of 0.16 to 0.70% by weight.

[0082] In an embodiment, the compounds may comprise herbal component with function similar to Withanolide A in an amount of 1.20 to 6.00% by weight.

[0083] In an embodiment, the compounds may comprise herbal component with function similar to Withaferin A in an amount of 0.5 to 3.6% by weight.

[0084] In an embodiment, the compounds may comprise herbal component with function similar to Withanolide D in an amount of 0.51 to 2.40% by weight.

[0085] In an embodiment, the compounds may comprise herbal component with function similar to P-Sitosterol in an amount of 1.2 to 3.70% by weight.

[0086] In an embodiment, the compounds may comprise herbal component with function similar to Diosgenin in an amount of 1.30 to 6.10% by weight.

[0087] In an embodiment, the compounds may comprise herbal component with function similar to Arjunolic Acid in an amount of 0.15 to 1.20% by weight.

[0088] In an embodiment, the compounds may comprise herbal component with function similar to Caffeic Acid in an amount of 0.02 to 1.40% by weight.

[0089] In an embodiment, the compounds may comprise herbal component with function similar to Kaempferol in an amount of 0.20 to 1.30% by weight.

[0090] The compounds are in orally consumable form selected from the group consisting of granules, tablets, pills, capsules, pellets, syrups, powders, emulsions, and dispersions.

[0091] The compounds are useful in a therapeutic or prophylactic method for treating a disease on hypertension, obesity, arteriolosclerosis, atherosclerosis, and / or diabetes mellitus.

[0092] The compounds are useful for the treatment of diseases like metabolic disorders, obesity, hyperlipidemia, dyslipidemia, hypertriglyceridemia, hypoalphalipoproteinemia, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy diabetic cataracts, hypertension, coronary artery disease, heart failure, atherosclerosis and pulmonary arterial hypertension. The polyherbal compound based formulation of the present invention can be effectively utilized as an adjuvant in combination with both chemical and non-chemical antihypertensive and diuretic agents, such as, but not limited to, Captopril, Bostean, Enalapril, and Spironolactone. In an embodiment, the said combinations target specific receptors, for exampleB5156-00012

[0093] Aldosterone (Spironolactone) and ACE (Bostean) receptors, modulating key pathways involved in regulating hypertension. The synergistic effects of the said combinations not only enhance the primary receptor-targeting action but also contribute to broader off-target effects by influencing additional receptors implicated in hypertension’s pathophysiology. This multitarget approach results in a more comprehensive and potent therapeutic strategy for managing hypertension.

[0094] The active compounds in the claimed polyherbal compound based formulation may be extracted from various plant sources, including: Dioscin which can be derived from Dioscorea species such as, but not limited to, D. opposita, D. villosa, D. zingiberensis, D. batatas, D. esculenta; Withanolides which can be derived from Withania species such as, but not limited to, W. somnifera, W. coagulans, W. aristata, W. obtusifolia; P-Sitosterol which can be derived from plant sources such as, but not limited to, Zea mays, Glycine max, Trigonella foenum-graecum, Cucurbita pepo, Corylus avellana, Pinus species; Diosgenin which can be derived from plant sources such as, but not limited to, D. zingiberensis, D. villosa, D. alata, D. esculenta, Trigonella foenum-graecum; Arjunolic Acid which can be derived from Terminalia species such as, but not limited to, T. arjuna, T. bellerica, T. chebula, Carya ovate; Caffeic Acid which can be derived from plant sources such as, but not limited to, Coffea arabica, Echinacea purpurea, Cichorium intybus, Ocimum basilicum, Salvia officinalis, Lamiaceae family; and Kaempferol which can be derived from plant sources such as, but not limited to, Camellia sinensis, Ginkgo biloba, Brassica oleracea, Allium cepa, Apteryx mantelli, Passiflora incarnata, Spinacia oleracea. These compounds may be extracted using standard processes such as Solvent Extraction or Ultrasonic-Assisted Extraction, or may be commercially procured. In an embodiment, the process of obtaining purified extract of the polyherbal compounds for the management of hypertension and related disorders comprising the steps of:

[0095] Step 1: The active ingredients are extracted from herbal sources using a standard extraction process such as, but not limited to, Solvent Extraction or Ultrasonic-Assisted Extraction, wherein the extraction is performed by the hydro-alcoholic solvent comprising 50-80 % alcohol such as, but not limited to, ethanol, methanol, isopropanol, etc. with 50-20% water, by soaking the grinded powder sieve through mesh # 40-60 of the herbs in the hydro-alcoholic solvent for 48-72 hours with occasional stirring at 50-80 RPM for 15-30 minutes and sonicated at 30-40°C for 20-40 minutes; filtration followed by, solvent evaporation using suitable method such as, but not limited to rotary evaporator, spray dryer, tray dryer, etc.B5156-00012

[0096] Step 2: The extracts are then purified to remove impurities and non-active compounds, through techniques such as, but not limited to, liquid chromatography, flash chromatography, column chromatography or recrystallization.

[0097] Step 3: The purified extracts are standardized to a specific concentration and, confirmed through analytical methods such as, but not limited to, HPLC, HPTLC UV-Vis spectrophotometry, gas-chromatography or mass spectrometry, etc. This ensures that the final purified extract contains desired active compounds within the specified ranges.

[0098] In another embodiment, a process of separation and collection of purified extract fractions / compounds (PEc) using column chromatography, comprising the steps of:

[0099] Step 1: Column Preparation

[0100] • Packing the column of appropriate size based on the amount of sample and desired resolution with bubble free stationary phase such as, but not limited to, silica gel, alumina, or other specialized materials either manually or using a slurry technique. Step 2: Sample Preparation

[0101] • Dissolving sample in a small amount of an appropriate solvent(s) with suitable polarity that should be compatible with the mobile phase and filter it to remove any insoluble impurities using a suitable filtration techniques.

[0102] Step 3: Conditioning the Column

[0103] • Run a small volume of the mobile phase through the column to equilibrate the stationary phase and ensure that column is free from air bubbles for uniform flow. The said process is done at temperature ranging between 20-35°C at relative humidity of 35 to 65.

[0104] Step 4: Loading the sample / extract

[0105] • Carefully loading the dissolve and filter sample / extract onto the top of the column without disturbing the stationary phase, and wait for some time to completely absorb the sample / extract in to the stationary phase for efficient separation.

[0106] Step 5: Elution

[0107] Carefully addition of the mobile phase (solvent mixture with suitable polarity) at the top of the column depending on the polarity and properties of the compounds being separated and by maintaining optimum flow rate of mobile phase (solvent) with suitableB5156-00012

[0108] techniques such as, but not limited to, gravity or a pump for uniform flow through the column, to get better separation of the Purified Extract compounds (PEc).

[0109] Step 6: Collection of Fraction / Compounds

[0110] • Collect Purified Extract Fractions / Compounds (PEc) by carefully monitoring the elution by checking for color changes or by using suitable techniques such as, but not limited to, Peak / band identification, Continuous fraction collection, Gradient elution with fraction collection, Manual fraction collection, Elution with constant volume, Elution with constant flow rate, etc. As the mobile phase moves down the column collect eluted fractions of compounds in the separate clean collecting vessels, such as but not limited to, vials, test tubes, beakers, etc. and label them appropriately evaporate the solvent using different techniques, such as but not limited to, rotary evaporator, vacuum dryer, spray dryer, nitrogen evaporator, etc.

[0111] Step 7: Identification of Fractions / Compounds

[0112] • Identification of Fractions / Compounds (PEc) using High Performance Thin Layer Chromatography (HPTLC), HPLC, MS, GC, UV spectrophotometry for the presence of separated compounds / fractions.

[0113] Step 8: Mixing and Homogenization:

[0114] • Mixing of separated / isolated compounds / fractions (PEc) by combining them in above mentioned ranges and homogenization.

[0115] In another embodiment, the process of preparation the polyherbal compound based solid formulation for the management of hypertension and related disorders comprises the steps of: Step 1: Pass the purified extract of desired active compounds (PEc) and diluents separately through a sieve with a mesh size of #40-60 and mix uniformly for 5-15 minutes at 50-150 rpm in mixing vessel such as, but not limited to mass mixer, blenders, tumblers, etc.

[0116] Step 2: Prepare the granulating fluid by dissolving preservatives and binders as mentioned in below embodiments into a suitable solvent under stirring to form a lump-free solution.

[0117] Step 3: Add the granulating fluid from Step 2 to the mixture from Step 1 to obtain a wet mass with suitable consistency.

[0118] Step 4: Dry the wet mass at a temperature between 65-70°C, with intermediate racking, until the loss on drying (LOD) is between 3.00-5.00% w / w.B5156-00012

[0119] Step 5: Pass the dried material through a sieve with a mesh size of #20-40 and mix it uniformly with a suitable diluent as mentioned in below embodiments at 20-50 rpm in a mixing vessels such as, but not limited to, mass mixer, various types of blenders, etc.

[0120] Step 6: As per requirement of different oral dosage forms this mixture can be used to formulate capsule, compressed into tablet or powder for suspension, emulsion, etc. as per known art. In an embodiment, a process for preparing poly herbal compound based liquid formulations, comprising the steps of:

[0121] STEP 1: Weighing the PEc and dispersing them into water at 60-70°C, stirring to form a uniform decoction, then cooling and filtering through a #80-100 mesh.

[0122] STEP 2: Dissolving preservatives as mentioned in below embodiments into purified water, heating the mixture to 80-90°C, and allowing it to cool to room temperature.

[0123] STEP 3: Combining the cooled decoction and preservatives dissolved purified water in a mixing vessel and mix thoroughly to get homogenized uniform mixture.

[0124] STEP 4: Adding additional ingredients such as taste enhancers and solvents as mentioned in below embodiments and then mixing well.

[0125] STEP 5: Adjusting the volume by adding purified water and ensuring the final mixture is uniform.

[0126] In an embodiment, the diluents used in the preparation process of the said invention are natural diluents such as, but not limited to, microcrystalline cellulose, corn starch, anhydrous lactose, Dextrates, lactose monohydrate, Dextrose Dextrin, Maltodextrin and Xylitol.

[0127] In an embodiment, the preservatives used in the preparation process of the claimed invention are natural preservatives such as, but not limited to, Tocopherol, Benzyl Alcohol, Citric Acid, Ginger Extract and Garlic Extract.

[0128] In an embodiment, the binders used in the preparation process of the claimed invention are natural binders such as, but not limited to, Starch, Sucrose, Acacia Gum, Guar Gum, Xanthan Gum and Pectin.

[0129] In an embodiment, the lubricants used in the preparation process of the claimed invention are natural lubricants such as Stearic Acid, Fumaric acid, Glyceryl behenate, and Glyceryl palmito stearate.B5156-00012

[0130] In an embodiment, the glidants used in the preparation process of the claimed invention are natural glidants such as, but not limited to, Silicon Dioxide, Rice Starch and Talc.

[0131] In an embodiment, the taste enhancers used in the preparation process of the claimed invention are natural taste enhancers such as, but not limited to, Vanillin, Mint, Rose, Honey, Saffron, etc.

[0132] In an embodiment, the solvent used in the preparation process of the claimed invention includes, but not limited to, aqueous, non-aqueous, polar, non-polar, alcoholic and hydroalcoholic solvents, wherein the alcohol ranges from 20% -70% in hydro-alcoholic solvent. The Invention is further described with the help of non-limiting experiments, as described below:

[0133] Example 1:

[0134] Extraction and Formulation

[0135] The extraction of the desired phytochemicals from the processed plant materials (Root, leaf, Bark etc.) was performed using hydro alcoholic solvent system.

[0136] The purified extract (PEc) obtained from the above extraction was used to develop the formulation C 1224c using the standard process known in art.

[0137] TABLE 1: Compounds of Formula C1224c:

[0138]

[0139] B5156-00012

[0140] MULTIPLE MECHANISTIC PATHWAY OF THE COMPOUNDS:

[0141] Hypertension is driven by a combination of overactive RAAS, sympathetic nervous system (P-AR), impaired vasodilation (NO and sGC pathways), increased vascular resistance (Endothelin-1), and fluid retention. No single pathway accounts for all aspects of high blood pressure, so inhibiting multiple mechanisms provides more comprehensive control.

[0142] ACE (Angiotensin-Converting Enzyme): Converts angiotensin I to angiotensin II, a potent vasoconstrictor that raises blood pressure. ACE inhibitors are common medications used to treat hypertension.

[0143] Angiotensin II receptor type 1 (ATI): This receptor binds angiotensin II, leading to vasoconstriction and aldosterone release, which increases blood pressure. Angiotensin II receptor blockers (ARBs) are used to block this pathway and lower blood pressure.

[0144] P-AR (Beta-Adrenergic Receptors): These receptors, when stimulated by adrenaline, increase heart rate and contractility, leading to higher blood pressure. Beta-blockers are used to reduce these effects and treat hypertension.

[0145] CACNB1 (Calcium Channel, Voltage-Dependent, Beta 1 Subunit): This gene encodes a subunit of voltage-gated calcium channels, involved in regulating calcium influx in cardiac and smooth muscle cells. Calcium channel blockers reduce blood pressure by inhibiting calcium entry.

[0146] Aldosterone: A hormone that increases sodium and water retention, which raises blood volume and blood pressure. Drugs that block aldosterone, such as aldosterone antagonists, are used to manage hypertension.

[0147] Endothelin-1: A potent vasoconstrictor that plays a significant role in blood vessel constriction and elevated blood pressure. Endothelin receptor antagonists are sometimes used in hypertension management.

[0148] Neprilysin: An enzyme that degrades natriuretic peptides, which are involved in lowering blood pressure. Inhibitors of neprilysin, in combination with ARBs, are used to treat hypertension and heart failure (e.g., sacubitril / valsartan).

[0149] NOS3 (Nitric Oxide Synthase 3): Produces nitric oxide, a molecule that causes vasodilation, reducing blood pressure. Impaired nitric oxide production is associated with hypertension. Renin: An enzyme that converts angiotensinogen to angiotensin I, initiating the RAAS pathway that increases blood pressure. Renin inhibitors, like aliskiren, are used to treat hypertension.B5156-00012

[0150] SESN2 (Sestrin 2): A stress-inducible protein that plays a role in regulating cellular metabolism and oxidative stress. Its role in hypertension is being studied, potentially related to oxidative stress management in cardiovascular diseases.

[0151] sGC (Soluble Guanylate Cyclase): Produces cyclic GMP (cGMP), which induces vasodilation. sGC stimulators and activators can reduce blood pressure in conditions like pulmonary hypertension.

[0152] ADMA (Asymmetric Dimethylarginine): An endogenous inhibitor of nitric oxide synthase, contributing to reduced nitric oxide production and thus vasoconstriction, which can increase blood pressure. Elevated ADMA levels are associated with hypertension.

[0153] By inhibiting several pathways at once, the therapeutic effects are often synergistic, resulting in greater blood pressure reduction than targeting a single pathway. For instance, combining RAAS inhibition (ACE or ARBs) with P-blockers and calcium channel blockers provides better outcomes in many patients with complex hypertension.

[0154] The body compensates for the inhibition of one pathway by activating others. For example, blocking ACE can lead to increased renin activity, which might negate some benefits. By simultaneously inhibiting renin or aldosterone, you prevent these compensatory mechanisms, improving treatment efficacy.

[0155] Long-term hypertension leads to damage to the heart, kidneys, and blood vessels. Multi-target therapy not only lowers blood pressure but also reduces end-organ damage by addressing multiple harmful mechanisms, such as oxidative stress (SESN2) and vascular remodelling (endothelin-1).

[0156] TABLE 2: Binding Affinity & Inhibition Constant of the individual active compounds in the Poly herbal Compounds.

[0157]

[0158] B5156-00012

[0159]

[0160] Compounds with hydroxyl (-OH) groups, such as Withaferin A, Withanolide D and Kaempferol exhibit very low IC50 values. This suggests that hydroxyl groups may enhance binding affinity by facilitating hydrogen bonding with target proteins.

[0161] Compounds featuring polycyclic structures, such as Withaferin A and Dioscin, tend to have lower IC50 values (2.27 nM and 7.01 nM). The rigidity and spatial arrangement provided by these structures may contribute to better fit within the active sites of target proteins in comparison to structures with fewer rings such as gallic acid.

[0162] Compounds that interact strongly with key residues such as ASN, TRP, and LYS often exhibit lower IC50 values, suggesting that these interactions are crucial for effective binding Dioscin, Withanolide D, Withaferin A and Withanolide D demonstrated the strongest binding affinities, respectively, indicating their potential as effective inhibitors.

[0163] 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).

[0164] 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:

[0165] Caffeic acid (MW: 180.16, LogP: 1.15, HBA: 4, HBD: 3, Bioavailability Score: 0.56) met all parameters without any violations.

[0166] Kaempferol (MW: 286.23, LogP: 2.28, HBA: 6, HBD: 4, Bioavailability Score: 0.55) adhered to RO5 with no violations.B5156-00012

[0167] 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.

[0168] TABLE 3: Drug Likeness Properties and Bioavailability Score of Selected Ligands

[0169]

[0170] In terms of safety, the phytochemicals were evaluated for their LD50, hepatotoxicity, carcinogenicity, and mutagenicity. The data highlights compounds with favorable safety profiles, such as:

[0171] The evaluated phytochemicals exhibit diverse safety and toxicity profiles, with most demonstrating no hepatotoxicity, carcinogenicity, or mutagenicity on the basis of in vitro studies except Caffeic acid. Compounds like Kaempferol, P-Sitosterol, and Diosgenin are particularly safe, with high LD50 values (>800 mg / kg), indicating low acute toxicity. Withanolide A (LD50: 34 mg / kg) and Dioscin (LD50: 55 mg / kg) have narrow safety margin but are notable for their bioactivity and therapeutic potential. These findings support the use of these phytochemicals in formulations, balancing efficacy with safety considerations.B5156-00012

[0172] TABLE 4: Toxicity prediction of selected phytochemicals compounds

[0173]

[0174] The docking results against Aldosterone revealed P-sitosterol as the ligand with the strongest binding affinity (-11.46 kcal / mol), interacting with key residues such as PHE D: 445, CYS D: 450, and LEU D: 380. Other highly potent compounds included Withaferin A (-11.31 kcal / mol), Withanolide A (-11.18 kcal / mol), and Arjunolic Acid (-11.14 kcal / mol), which formed interactions with residues like PHE D: 487, ARG D: 384, and PHE D: 130. Diosgenin (-10.58 kcal / mol) and the standard drug Spironolactone (-10.58 kcal / mol) also exhibited strong binding. Moderate binding affinities were observed for Withanolide D (-10.65 kcal / mol) and Dioscin (-7.9 kcal / mol), while Caffeic acid and Kaempferol showed weaker binding (-6.8 and -7.2 kcal / mol, respectively). Residues such as CYS D: 450, PHE D: 487, and ARG D: 384 were consistently involved, indicating their importance in ligand binding. These findings suggest P-sitosterol and Withaferin A as highly promising candidates for Aldosterone receptor inhibition.B5156-00012

[0175] TABLE 5: Docking results of selected compounds against Aldosterone

[0176]

[0177] B5156-00012

[0178]

[0179] The docking study against Neprilysin identified Dioscin as the compound with the highest binding affinity (-11.2 kcal / mol), interacting with residues such as SER A: 547, GLN A: 521, and VAL A: 710. Withanolide A (-10.41 kcal / mol) and Withaferin A (-10.08 kcal / mol) also exhibited strong binding, forming interactions with critical residues like HIS A: 711, TRP A: 693, and PHE A: 106. Withanolide D (-9.61 kcal / mol) and P-sitosterol (-9.54 kcal / mol) showed moderate binding, while the standard drug Sacubitril had a binding affinity of -8.03 kcal / mol. Diosgenin (-5.28 kcal / mol) and Kaempferol (-5.9 kcal / mol) demonstrated weaker binding. Key residues such as HIS A: 711, HIS A: 583, TRP A: 693, and GLU A: 584 were commonly involved, emphasizing their importance in ligand binding. These findings suggest Dioscin, Withanolide A, and Withaferin A as potent inhibitors with potential therapeutic applications targeting Neprilysin.

[0180] TABLE 6: Docking results of selected compounds against Neprilysin

[0181]

[0182] B5156-00012

[0183]

[0184] The docking study of selected compounds against ACE revealed varied binding affinities and amino acid interactions, demonstrating their potential as ACE inhibitors. Withaferin A and Withanolide D exhibited the highest binding affinities (-11.79 kcal / mol and -11.75 kcal / mol, respectively), interacting with key residues such as ASN A: 210, LYS A: 562, and TRP A: 566. Dioscin also showed strong binding (-11.00 kcal / mol), engaging residues like PRO A: 346 and TRP A: 349. Moderate binding affinities were observed for P-sitosterol (-10.87 kcal / mol) and Withanolide A (-10.76 kcal / mol), while Caffeic acid and Captopril had the lowest binding affinities (-6.13 kcal / mol and -6.04 kcal / mol, respectively). Key residues involved across compounds included LEU A: 95, VAL A: 209, and PRO A: 565, suggesting their importance in ACE inhibition. These results highlight the potential of phytochemicals, particularly Withaferin A and Withanolide D, as promising ACE inhibitors for therapeutic applications.B5156-00012

[0185] TABLE 7: Docking results of selected compounds against ACE

[0186]

[0187] B5156-00012

[0188] The docking study of selected compounds against Endothelin- 1 demonstrated varying binding affinities and interactions with critical amino acid residues. Dioscin exhibited the highest binding affinity (-6.8 kcal / mol), interacting with key residues such as SER A: 2, CYS A: 3, and PHE A: 14. Withanolide A (-6.76 kcal / mol) and Withanolide D (-6.55 kcal / mol) also showed strong affinities, forming interactions with residues like CYS A: 3, MET A: 7, and ILE A: 19. Moderate affinities were observed for Diosgenin (-6.22 kcal / mol) and Arjunolic Acid (-6.36 kcal / mol), while standard drug Bosentan had a lower affinity (-5.08 kcal / mol). Commonly interacting residues included CYS A: 3, CYS A: 15, ILE A: 19, and HIS A: 16, indicating their crucial role in ligand binding. These findings suggest that Dioscin, Withanolide A, and Withanolide D have promising potential as Endothelin- 1 inhibitors.

[0189] TABLE 8: Docking results of selected compounds against Endothelin 1

[0190]

[0191] B5156-00012

[0192]

[0193] The docking results against SESN2 identified Dioscin as the compound with the strongest binding affinity (-8.7 kcal / mol), interacting with residues such as ASP A: 104, GLY B: 105, and LEU A: 362. Withaferin A (-7.66 kcal / mol) and Withanolide A (-7.53 kcal / mol) also demonstrated strong binding, engaging residues like HIS A: 132, ASP A: 77, and TRP A: 94. Withanolide D (-6.94 kcal / mol) exhibited moderate binding, while P-sitosterol (-5.4 kcal / mol) and Carnosol (-5.94 kcal / mol) showed weaker affinities. Caffeic acid had the lowest binding affinity (-5.2 kcal / mol), interacting with residues such as GLU A: 135 and HIS A: 113. Common interacting residues, including HIS A: 97, HIS A: 132, and ASP A: 77, highlight their significance in SESN2 binding. These results suggest Dioscin, Withaferin A, and Withanolide A as promising candidates for SESN2 modulation.

[0194] TABLE 9: Docking results of selected compounds against SESN2

[0195]

[0196] B5156-00012

[0197]

[0198] The docking study against NOS3 identified Dioscin as the top-performing compound with the highest binding affinity (-12.1 kcal / mol) and extensive interactions with key amino acids, including GLU A: 361, ARG A: 372, CYS A: 184, and PHE A: 353, indicating strong and stable binding. Other compounds such as Withanolide A (- 11.79 kcal / mol), P-sitosterol (-11.69 kcal / mol), Diosgenin (-11.60 kcal / mol), and Arjunolic Acid (-11.38 kcal / mol) also demonstrated robust binding, engaging critical residues like CYS A: 184 and VAL A: 336, essential for NOS3 activity. Moderate binding affinities were observed for Withanolide D (-10.54 kcal / mol) and Kaempferol (-6.33 kcal / mol), while Caffeic acid showed the weakest interaction (-5.18 kcal / mol). Notably, residues such as CYS A: 184, PHE A: 353, VAL A: 336, and TRP A: 178 appeared consistently across ligands, underscoring their importance in ligand stabilization. These results highlight Dioscin and similar high- affinity compounds as promising candidates for NOS 3 -targeted therapies

[0199] TABLE 10: Docking results of selected compounds against NOS3

[0200]

[0201] B5156-00012

[0202]

[0203] TABLE 11: Binding Affinity and Inhibition Constants for Aldosterone Ligand Interactions of the compounds as Adjuvant.

[0204]

[0205] B5156-00012

[0206] Kaempferol + Spironolactone exhibit a synergistic effect, significantly improving binding affinity (from -7.2 kcal / mol to -11.25 kcal / mol) and reducing the inhibition constant (from 5.27 pM to 5.67 nM). This indicates a highly potent interaction, indicating the combination's strong potential as a therapeutic agent targeting Aldosterone-related pathways.

[0207] Example 2:

[0208] TABLE 12: Binding Affinity and Inhibition Constants for Endothelin-1 Ligand Interactions of the compounds as Adjuvant.

[0209]

[0210] Bosentan + Withanolide A demonstrate an enhanced effect compared to Bosentan alone, with improved binding affinity (from -5.08 kcal / mol to -5.63 kcal / mol) and a reduction in inhibition constant (from 187.94 pM to 74.63 pM). While the improvement is moderate, it suggests potential for optimized activity against Endothelin-1 pathways.

[0211] Example 3:

[0212] IN VIVO EFFICACY STUDIES STUDY 1 (DOCA induced Hypertension)

[0213] The DOCA salt model is employed to create secondary hypertension in rats through the oral administration of DOCA and replacing the drinking water with 1% saline. This process activates the renin-angiotensin-aldosterone system (RAAS) and leads to increased sodium retention. The study encompasses four experimental groups: Normal Control, which receives standard care and serves as a baseline; Disease Control, where hypertension is induced without any treatment; Chemical Control, which is administered a standard antihypertensive drug to assess its impact on blood pressure; and Compounds C 1224c, aimed at evaluating the effectiveness of a test formulation in reducing hypertension. Blood pressure measurements (systolic, diastolic, and mean arterial pressure) were taken on Day 0 and Day 42 to monitor theB5156-00012

[0214] progression of hypertension and assess the effectiveness of the treatments. The results are shown in Figure 5a to 5g.

[0215] The disease control group exhibited substantial and uncontrolled increases in systolic, diastolic, and mean arterial pressures over 42 days, indicating significant progression of hypertension without treatment. Conversely, the chemical control group demonstrated remarkable efficacy in stabilizing blood pressure, with only minor increases observed across all parameters. Similarly, the C 1224c treatment group showed significant effectiveness in managing hypertension, demonstrating a marked reduction in blood pressure compared to the disease control group. Notably, the chemical control and C 1224c formulations were both highly effective, showing comparable efficacy in mitigating blood pressure elevations relative to the disease control group.

[0216] The Disease Control group exhibited significantly elevated levels of cholesterol, triglycerides, and LDL, accompanied by reduced HDL, indicating a high risk of cardiovascular complications. The Chemical Control group demonstrated the ability to regulate these lipid markers effectively, bringing them closer to normal levels with regular administration. The Compounds C 1224c treatment group showed equal efficacy in controlling lipid abnormalities and hypertension, achieving results comparable to the Chemical Control group and significantly superior to the Disease Control group.

[0217] Example 4:

[0218] STUDY 2 (L-NAME induced Hypertension)

[0219] 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 a standard antihypertensive drug to assess its effect on lowering blood pressure; and Compounds C1224c, a novel treatment aimed at evaluating its potential antihypertensive effects. Blood pressure (systolic, diastolic, and mean arterial pressure) is measured at Day 42 to assess the efficacy of each treatment. The results are shown in Figure 6a to 6g.

[0220] The disease control group exhibited substantial and uncontrolled increases in systolic, diastolic, and mean arterial pressures over 42 days, indicating significant progression of hypertension without treatment. Conversely, the chemical control group demonstrated remarkable efficacyB5156-00012

[0221] in stabilizing blood pressure, with only minor increases observed across all parameters. Similarly, the C 1224c treatment group showed significant effectiveness in managing hypertension, demonstrating a marked reduction in blood pressure compared to the disease control group. Notably, the chemical control and C 1224c formulations were both highly effective, showing comparable efficacy in mitigating blood pressure elevations relative to the disease control group.

[0222] The Disease Control group exhibited significantly elevated levels of cholesterol, triglycerides, and LDL, accompanied by reduced HDL, indicating a high risk of cardiovascular complications. The Chemical Control group demonstrated the ability to regulate these lipid markers effectively, bringing them closer to normal levels with regular administration. The Compounds C 1224c treatment group showed equal efficacy in controlling lipid abnormalities and hypertension, achieving results comparable to the Chemical Control group and significantly superior to the Disease Control group.

Claims

B5156-00012We Claim:

1. Polyherbal compound based formulation for the management of hypertension and related disorders comprising:a) Dioscin present in an amount ranging from 0.16 to 0.70%;b) Withanolide A present in an amount ranging from 1.20 to 6.00%;c) Withaferin A present in an amount ranging from 0.5 to 3.6%;d) Withanolide D present in an amount ranging from 0.51 to 2.40%;e) P-Sitosterol present in an amount ranging from 1.2 to 3.70%;f) Diosgenin present in an amount ranging from 1.30 to 6.10%;g) Arjunolic Acid present in an amount ranging from 0.15 to 1.20%;h) Caffeic Acid present in an amount ranging from 0.02 to 1.40%;i) Kaempferol present in an amount ranging from 0.20 to 1.30%.

2. The compounds as claimed in claim 1, wherein said disorder comprises hypertension and vascular stress, and other metabolic disorders, obesity, hyperlipidemia, dyslipidemia, hypertriglyceridemia, hypoalphalipoproteinemia, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy diabetic cataracts, coronary artery disease, heart failure, atherosclerosis and pulmonary arterial hypertension.

3. The compounds as claimed in claim 1, wherein the said compounds are useful as a nutritional supplement or as an adjuvant in combination with at least one antihypertensive agent.

4. The compounds as claimed in claim 1, wherein said compounds targets multiple mechanistic pathways5. A method of preparing a polyherbal compound based solid formulation for the management of hypertension and other disorders comprising the steps of:a) Sifting purified extract of desired active compound (PEc) and diluents through a #40-60 mesh sieve, and mixing them uniformly for 5-15 minutes at 50-150 rpm in mixing vessel.b) Preparing a granulating fluid by dissolving preservatives and binders into a suitable solvent under stirring to form a lump-free solution.B5156-00012c) Adding the granulating fluid to the mixture from Step 2 to form a wet mass with suitable consistency.d) Drying the wet mass at a temperature of 65-70°C with intermediate racking until the loss on drying (LOD) is between 3.00-5.00% w / w.e) Sifting the dried mass through a #20-40 mesh sieve and mixing it uniformly with a suitable diluent at 20-50 rpm in mixing vessels such as, but not limited to, mass mixer, various types of blenders, etc.f) The mixture further comprises, according to requirement of dosage form such as powder, granules, pellets, tablet, capsule, suspension, etc.

6. A method of preparing a poly herbal compound based liquid formulation for the management of hypertension and other disorders comprising the steps of:a) Weighing the PEc and dispersing them into water at 60-70°C, stirring to form a uniform decoction, then cooling and filtering through a #80-100 mesh. b) Dissolving preservatives into purified water, heating the mixture to 80-90°C, and allowing it to cool to room temperature.c) Combining the cooled decoction and preservatives dissolved purified water and mixing thoroughly to get homogenized uniform mixture.d) Adding additional ingredients such as taste enhancers and solvents, then mixing well.e) Adjusting the volume by adding purified water and ensuring the final mixture is uniform.

7. A method of obtaining extract of the herbs for the compounds as claimed in claim 1, comprising the steps of:a) Extracting the active ingredients from herbal sources using Solvent Extraction, or Ultrasonic-Assisted Extraction, wherein the extraction is performed by the hydro-alcoholic solvent comprising 50-80 % alcohol such as, but not limited to, ethanol, methanol, isopropanol, etc. and 50-20% water, by soaking the grinded powder sieve through mesh # 40-60 of the herbs in the hydro-alcoholic solvent for 48-72 hours with occasional stirring at 50-80 RPM for 15-30 minutes and sonicated at 30-40°C for 20-40 minutes; filtration followed by, solventB5156-00012evaporation using suitable method such as, but not limited to rotary evaporator, spray dryer, tray dryer, etc.b) Purifying the extracts through liquid chromatography, flash chromatography, column chromatography, or recrystallization to remove impurities;c) Standardizing the purified extracts to a specific concentration and confirmed through analytical methods, such as but not limited to, HPTLC, HPLC, UV-Vis spectrophotometry, or mass spectrometry, etc.

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