Description:TECHNICAL FIELD

[0001] The present invention relates to the holistic approach to managing lipid levels. More specifically, the inventions relate to an herbal approach as an alternative treatment option for drug therapy for treatment of hyperlipidemia.

BACKGROUND ART

[0002] Coronary heart diseases (CHD) are the main cause of death in western countries and Asia. Among CHDs, ischemic heart disease (IHD) leads to the highest mortality rate. The number of heart patients suffering from IHD worldwide is gradually increasing. In recent years much prominence has been given to the association of abnormal levels or values of lipid profile (e.g., total cholesterol, LDL, VLDL and triglycerides) with atherosclerosis and ischemic heart disease (IHD).

[0003] Treatment of hyperlipidemia is preferably by dietary factors accompanied by other natural regimes. Drug therapy is reserved for more intractable conditions. Thus, foremost in the development and management of atherosclerosis is the reduction of serum cholesterol levels. Individuals (men or women) with 33 to 44 years of age, the total cholesterol levels of 256 mg/dL or over have a 5 times greater risk of developing coronary artery disease than those whose levels are below 220 mg/dL.

[0004] While drug therapy for hyperlipidemia has proven to be effective in reducing cholesterol and triglyceride levels, it also comes with some disadvantages and potential drawbacks. Some of the main disadvantages of drug therapy in the treatment of hyperlipidemia are side effect, drug interaction, liver toxicity, muscle toxicity, cost, adherence challenge and limited effect

[0005] Thus, herbal medicines are being used by about 80% of the world population primarily in the developing countries for primary health care. Herbal medicine has stood the test of time for their safety, efficacy, cultural acceptability and lesser side effects. The chemical constituents present in them are a part of the physiological functions of living flora and hence they are believed to have better compatibility with the human body.

[0006] In order to overcome the aforementioned drawbacks, there is a need in the art to develop plant-based preparations of the herb to treat various health conditions, including hyperlipidemia.
OBJECTS OF THE INVENTION

[0007] The principal object of the present invention is to overcome the disadvantages of the prior art.
[0008] An object of the present invention is to develop an herbal formulation as therapy for hyperlipidemia.
[0009] Another object of the present invention is to provide a composition demonstrating lipid-lowering properties.
[0010] Yet another object of the present invention is to employ a cost-effective and eco-friendly composition.
[0011] The foregoing and other objects of the present invention will become readily apparent upon further review of the following detailed description of the embodiments as illustrated in the accompanying drawings.
SUMMARY OF THE INVENTION

[0012] The present invention relates to herbal composition comprising Salvadora oleoides that exhibit enhanced overall hyperlipidemia treatment outcome.
[0013] According to an embodiment of the present invention, the method for preparation of the composition comprises various steps. The method may include a step of collecting and cleaning Salvadora oleoides to remove dirt or debris, followed by drying to obtain a dried plant material. Further, the method may include a step of grinding the dried plant material using a mortar and pestle or a mechanical grinder to obtain a powder. It may include a step of soaking the powder in a solvent such as methanol, petroleum ether, chloroform, ethyl-acetate and butanol for a certain period to obtain an extract. It may include further a step of filtering the extract, followed by evaporation by rotary evaporator to obtain the herbal composition.
[0014] While the invention has been described and shown with reference to the preferred embodiment, it will be apparent that variations might be possible that would fall within the scope of the present invention.
BRIEF DESCRIPTION OF DRAWINGS

[0015] So that the manner in which the above-recited features of the present invention can be understood in detail, a more particular description of the invention, briefly summarized above, may have been referred by embodiments, some of which are illustrated in the appended drawings. It is to be noted, however, that the appended drawings illustrate only typical embodiments of this invention and are therefore not to be considered limiting of its scope, for the invention may admit to other equally effective embodiments.
[0016] These and other features, benefits, and advantages of the present invention will become apparent by reference to the following text figure, with like reference numbers referring to like structures across the views, wherein:
[0017] Figure 1 illustrates a flowchart for preparation of the herbal composition; and
[0018] Figure 2 illustrates a flowchart for column chromatography of butanol extraction.
DETAILED DESCRIPTION OF THE INVENTION

[0019] While the present invention is described herein by way of example using embodiments and illustrative drawings, those skilled in the art will recognize that the invention is not limited to the embodiments of drawing or drawings described and are not intended to represent the scale of the various components. Further, some components that may form a part of the invention may not be illustrated in certain figures, for ease of illustration, and such omissions do not limit the embodiments outlined in any way. It should be understood that the drawings and the detailed description thereto are not intended to limit the invention to the particular form disclosed, but on the contrary, the invention is to cover all modifications, equivalents, and alternatives falling within the scope of the present invention as defined by the appended claim.
[0020] As used throughout this description, the word "may" is used in a permissive sense (i.e. meaning having the potential to), rather than the mandatory sense, (i.e. meaning must). Further, the words "a" or "an" mean "at least one” and the word “plurality” means “one or more” unless otherwise mentioned. Furthermore, the terminology and phraseology used herein are solely used for descriptive purposes and should not be construed as limiting in scope. Language such as "including," "comprising," "having," "containing," or "involving," and variations thereof, is intended to be broad and encompass the subject matter listed thereafter, equivalents, and additional subject matter not recited, and is not intended to exclude other additives, components, integers, or steps. Likewise, the term "comprising" is considered synonymous with the terms "including" or "containing" for applicable legal purposes. Any discussion of documents, acts, materials, devices, articles, and the like are included in the specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention.
[0021] In this disclosure, whenever a composition or an element or a group of elements is preceded with the transitional phrase “comprising”, it is understood that we also contemplate the same composition, element, or group of elements with transitional phrases “consisting of”, “consisting”, “selected from the group of consisting of, “including”, or “is” preceding the recitation of the composition, element or group of elements and vice versa.
[0022] The present invention is described hereinafter by various embodiments with reference to the accompanying drawing, wherein reference numerals used in the accompanying drawing correspond to the like elements throughout the description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiment set forth herein. Rather, the embodiment is provided so that this disclosure will be thorough and complete and will fully convey the scope of the invention to those skilled in the art. In the following detailed description, numeric values and ranges are provided for various aspects of the implementations described. These values and ranges are to be treated as examples only and are not intended to limit the scope of the claims. In addition, several materials are identified as suitable for various facets of the implementations. These materials are to be treated as exemplary and are not intended to limit the scope of the invention.
[0023] The present invention pertains to an herbal therapy for treatment of hyperlipidemia that exhibits minimal side effects and interactions with other medications, thereby replacing conventional medical treatments.
[0024] In one embodiment, the herbal composition comprises i) Salvadora oleoides and ii) solvent. The solvent may include but not limited butanol, ethanol, methanol, petroleum ether, chloroform, ethyl-acetate and alike.
[0025] In another embodiment, the herbal composition may comprise of additives selected from a group of an emulsifying agent, surface active agent, antimicrobial, antibacterial agent, essential oil and alike.
[0026] In one embodiment, the method for preparing the herbal composition having anti-hyperlipidemic activity is conducted via cold percolation. The method comprises various steps. The method may include a step of collecting and cleaning Salvadora oleoides to remove dirt or debris, followed by drying to obtain a dried plant material. The method further may comprise a step of grinding the dried plant material using a mortar and pestle or a mechanical grinder to obtain a powder. The method may include a step of soaking the powder in a solvent for a certain period to obtain an extract. Further, the method may include a step of filtering the extract, followed by evaporation by rotary evaporators to obtain the herbal composition.
[0027] For better understanding, the developed composition comprising solvent extract of the Salvadora oleoides was further characterized.
Acute toxicity studies
[0028] Acute toxicity studies were carried out on albino mice. The composition administered orally/intraperitoneally in graduated doses to several groups of experimental animals, whereas one dose is being used per group. The compound administered once, orally or parenterally, to rats that have been fasted for 18 hrs.
[0029] Albino mice of 10 animals per group and weighing 20-25 g were administered graded dose (100-3000 mg/kg body weight, i.p.) of the total methanol extract and its fractions of Salvadora oleoides (decne).
[0030] After administration of the different fractions the mice were observed for toxic effects after 48hr of treatment. The toxicological effects were observed in terms of mortality expressed as LD50. The number of animals dying during a period was noted. The LD50 of the fractions was determined by OECD 401 guidelines.
[0031] It was concluded that Salvadora oleoides methanol, its fractions and compound SOD2 did not produce any toxic symptoms and mortality when administered orally to mice at a dose of 3000 mg/kg body weight in albino mice.
Column chromatography
[0032] Column chromatography is a widely used separation technique in chemistry and biochemistry to purify and separate components of a mixture based on their differential affinities for a stationary phase and a mobile phase. It is a form of liquid chromatography, where the mobile phase is a liquid solvent that flows through a column packed with a solid stationary phase.
[0033] Referring to figure 2, butanol fraction of plant was subjected to purification for isolation of active principle by column chromatography. Four sub-fractions obtained by column chromatography namely, Sub-fraction A, B, C and D based on TLC. All sub-fractions of plants were screened for antihyperlipidemic potential.
[0034] Butanol fraction of Salvadora oleoides showed significant reduction (p<0.05) in serum cholesterol (61.73 mg/dl), triglyceride (121.25 mg/dl), LDL (37.89 mg/dl), VLDL (24.98 mg/dl) and increase in HDL level (36.44 mg/dl) in comparison with standard drug fenofibrate (p<0.05).
[0035] Then active butanol fractions of the plant were subjected to column chromatography for the isolation of active principle. After purification by column chromatography four sub-fractions were obtained i.e., Sub-fraction A, B, C and D which were again evaluated for their antihyperlipidemic activity at a dose of 65 mg/kg (oral) to the Triton WR-1339 induced hyperlipidemic rats and total cholesterol, triglycerides, HDL, LDL and VLDL level in the blood were checked.
[0036] Sub-fraction D of Salvadora oleoides showed significant reduction in plasma cholesterol (73.97 mg/dl), triglyceride (94.29 mg/dl), LDL (21.38 mg/dl), VLDL (18.86 mg/dl) and increase in HDL level (33.95 mg/dl) in comparison with standard drug fenofibrate (p<0.05.
[0037] The active sub-fraction D of the plant was subjected to purification by column chromatography for the isolation compound Salvadoraside (SOD2) from Salvadora oleoides was obtained. The antihyperlipidemic potential in Salvadoraside (SOD2) from Salvadora oleoides showed significant reduction in plasma cholesterol (76.34 mg/dl), triglyceride (85.53 mg/dl), LDL (24.52 mg/dl), VLDL (17.71 mg/dl) and increase in HDL level (34.11 mg/dl) in comparison with standard drug fenofibrate. Thus, fraction D was found to possess good antihyperlipidemic potential.
Antihyperlipidemic activity
[0038] Hyperlipidemia was induced by single intraperitoneal dose of Triton WR 1339 (350mg/kg) and confirmed by elevated fasting serum Cholesterol level after 48 hrs of Triton dose. Total methanol extract and its fractions administered at a dose level of 100mg/kg body weight for 07 days. On the 7th day total methanol extract and its fractions reduced blood cholesterol level significantly (p<0.05).
[0039] Among all the fractions, butanol fraction from Salvadora oleoides caused the most significant reduction in serum cholesterol level. The active butanol fraction from Salvadora oleoides were subjected to purification by column chromatography in different solvent systems (mobile phase) in order to isolate the active component responsible for antihyperlipidemic activity in Salvadora oleoides respectively.
[0040] The purification by column chromatography led to isolation of four fraction based on TLC i.e., A, B, C, and D. They were screened for antihyperlipidemic potential at a dose of 65 mg/kg body weight in Triton WR 1339 induced hyperlipidemic rats for 07 days. After the 7th day ‘D’ of both plants showed the most significant reduction in serum cholesterol level as. Both active fractions were further purified which led to isolation of active principles.
[0041] These active constituents were named as Salvadoraside (SOD2) from Salvadora oleoides. The yield of these constituents was 5.5 % and 7.0% respectively. The isolated compound was investigated for their antihyperlipidemic effect. Isolated compounds showed significant reduction in serum cholesterol level of Triton WR 1339 induced hyperlipidemic rats at a dose of 50mg/kg.
[0042] Salvadoraside (SOD2) showed significant reduction in hyperlipidemia. The total methanol extract showed antihyperlipidemic activity at a dose level of 100mg/kg body weight and compared to that of the control group. The effect was comparable with that of standard drug fenofibrate (65 mg/kg body weight), a standard antihyperlipidemic drug.
[0043] Moreover, cholesterol and triglycerides levels were measured. Further antihyperlipidemic activity of its fraction i.e., Petroleum ether, Chloroform, Ethyl acetate and Butanol at a dose of 100mg/kg (oral) to the Triton WR-1339 induced hyperlipidemic rats were carried out.
[0044] Further, the operations need not be performed in the disclosed order, although in some examples, an order may be preferred. Also, not all functions need to be performed to achieve the desired advantages of the disclosed system and method, and therefore not all functions are required.
[0045] Various modifications to these embodiments are apparent to those skilled in the art from the description and the accompanying drawings. The principles associated with the various embodiments described herein may be applied to other embodiments. Therefore, the description is not intended to be limited to the 5 embodiments shown along with the accompanying drawings but is to be providing the broadest scope consistent with the principles and the novel and inventive features disclosed or suggested herein. Accordingly, the invention is anticipated to hold on to all other such alternatives, modifications, and variations that fall within the scope of the present invention and appended claims.
, Claims:I/We Claim:
1) An herbal composition having anti-hyperlipidemic activity comprises of
i) salvadora oleoides; and
ii) solvent.

2) The method as claimed in claim 1, wherein said solvent may include but not limited to methanol, petroleum ether, chloroform, ethyl-acetate, ethanol and butanol.

3) A method for preparing the herbal composition having anti-hyperlipidemic activity comprises the following steps:
i) Collecting and cleaning Salvadora oleoides to remove dirt or debris, followed by drying to obtain a dried plant material;
ii) Grinding said dried plant material using a mortar and pestle or a mechanical grinder to obtain a powder;
iii) Soaking said powder in methanol for a certain period to obtain an extract; and
iv) Filtering said extract, followed by evaporation, fractionation and isolation process as given below.

4) The method as claimed in claim 1, wherein said evaporation is carried out by rotary evaporators.