Annexure I

Brief description of the idea highlighting innovative element:

Development and Evaluation of Polyherbal Anti-hyperlipidaemic Oral Formulation

Containing four Indigenous Medicinal Plant Extracts

Hyperlipidemia refers to elevated lipid levels in blood. The condition is also called
hyperlipemia, dyslipidemia, lipemia, or lipidemia, and may be manifested by elevation of total
cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and total triglyceride (TG) levels
in the blood. Hyperlipidemia is one of the most complicated risk factor which ultimately
develops coronory syndrome and cardiovascular diseases (CVD) and because of its
complications like heart attack, stroke, ischemic organ disease including dementia, limb
gangrene are among the most leading causes of mortality.

According to ICMR–INDIAB Study, In India, there has been an alarming increase in the
prevalence of CVD over the past two decades so much so that accounts for 24% of all deaths
among adults aged 25–69 years. Hypercholesterolemia was found in 13.9%,
hypertriglyceridemia in 29.5%, low HDL-C in 72.3% and high LDL-C in 11.8% of the
population. Asian Indians have been found to develop CVD at a younger age than other
populations. The likely causes for the increase in the CVD rates include lifestyle changes
associated with urbanization and the epidemiologic and nutritional transitions that accompany
economic development. Dyslipidemia has been closely linked to the pathophysiology of CVD
and is a key independent modifiable risk factor for cardiovascular disease.

Proper recognition and management of dislipidemia can reduce cardiovascular and total
mortality rates. Current lipids modulating medications include bile-acid sequestrants, fibrates,
nicotinic acid, cholesterol absorption inhibitors, cholesteryl ester transfer protein inhibitors,
phytosterols, oil fish and HMG-CoA reductase inhibitors. Clinically, statins have been the most
widely prescribed drugs for hypercholesterolemia. Statins effectively lower the plasma
concentration of low density lipoprotein (LDL) cholesterol (LDL-C) and reduce mortality and
morbidity from CHD. However, many patients under statin treatment alone do not achieve the

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LDL-C goal suggested by the recent guidelines of the National Cholesterol Education Program’s

Adult Treatment Panel Ⅲ (ATP Ⅲ).

Many side effects retard the compliance of hypolipidemic drugs for treating
cardiovascular diseases. The bile acid sequestrants are not absorbed into the body so that they
don't have systemic side effects, but the most common effects include constipation, abdominal
pain, bloating, and dyspepsia. Niacin has been reported to cause severe liver toxicity and new
fibrates are known to be contraindicated in renal failure. fibrates in combination with statins
causes rhabdomyolysis.

Among the hypolipidemic drugs like bile acid sequestrants, nicotinic acid, fibric acid
derivatives, statins have lesser side effects in comparison but statins are also reported with
hepatic and renal failure. The Statins, though they are being used successfully for lowering the
lipid levels, will lose its power may be due to resistance or some other reasons in the coming
decade. It’s time to develop novel, effective and side-effect free poly-herbal formulation to treat
hyperlipidaemia.

Role of herbal drugs in the management of dyslipidaemia:

By the immense potential benefits of the medicinal plants which are being used from
years ago in curing many severe diseases may helpful to overcome the side effects of allopathic
drugs.

Traditionally, the herbal drugs like cinnamon bark, guggul, flaxseeds, fish oils,
fenugreek, garlic and tomatoes rich in phenols and flavonoids have been in use for the treatment
of hyperlipidaemia and scientifically all these crude drugs have been proved to have anti-
hyperlipidaemic activity on rat experiment models. So far, much research work has been carried
out on these herbs but no single formulation was developed or available in the market.

Based on the traditional uses and scientific work done on the herbal drugs, the crude
drugs Cinnamon bark, Arjuna bark, Fenugreek seeds and Guggul resin were selected for the
present project work.

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 The main aim of the project work is development of poly-herbal formulation and
screening of its lipid lowering activity. This work involves phytochemical screening and
biological evaluation of poly-herbal formulation.

Objectives of the experimental work to be carried out in the project work:

I. Steps involved in phytochemical screening of herbal drugs.

i) Procurement of plant materials (Cinnamon bark, Arjuna bark, Fenugreek
seeds and Guggul resin powders)
ii) Authentication of plant material by Botanist
iii) Drying of plant material
iv) Pulverization of plant material
v) Extraction of plant material using Soxhlet apparatus
vi) Preparation of crude extract.
vii) Preliminary Phytochemical screening of extract
viii) Development of poly-herbal formulations

II. The following models will be used for screening of anti-hyperlipidaemic activity of
poly-herbal formulations on rat experiment models.
i) Triton X 100 induced hyperlipidaemia model.
ii) High-fat diet-induced hyperlipidaemic study.
III. Submission of reports to the funding agency.

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Methodology:

I. Preparation of ethanol and aqueous extracts of Cinnamon bark, Arjuna bark, Fenugreek

seeds and Guggul resin powders):

i) Procurement of plant materials (Cinnamon bark, Arjuna bark,

Fenugreek seeds and Guggul resin powders):
All the plant materials will be purchased from local market if available,
otherwise can be purchased from YUCCA Enterprises, which is supplier of
crude drugs.
ii) Authentication of plant material by Botanist:
After collection of plant materials, they will be identified by
Dr.P.Satyanarayana Raju, Plant taxonomist, Dept of Botany and
Microbiology, Acharya Nagarjuna University, Guntur.
iii) Drying of plant material:
The plant materials that are collected will be dried under shade to prevent
the evaporation of volatile constituents from the crude drugs.
iv) Pulverization of plant material:
After drying, plant material will be subjected to pulverization to get
powder material.
v) Extraction of plant material using Soxhlet apparatus:
The powdered material of 1 kg of each of drugs Cinnamon bark, Arjuna
bark, Fenugreek seeds and Guggul resin powders will be successively
extracted with ethanol and water by the process of continuous soxhlet
extraction for 6 hr.
vi) Preparation of crude extract:
The crude extracts will be evaporated to dryness in a rotary film evaporator to
get dried extracts of different solvent.
vii) Preliminary Phytochemical screening of extract:
Extracts obtained in the extraction will be subjected to preliminary
phytochemical screening to identify the active compounds presented in the
ethanol and aqueous extracts of all the crude drugs.

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 viii) Development of poly-herbal oral formulations:
Solid dosage forms i.e., Poly-herbal oral formulations of different doses will
be prepared from all the extracts using wet granulation techniques.

II. Screening of anti-hyperlipidaemic activity of poly-herbal oral formulations on rat

experiment models:

i) Triton X 100 and High-fat diet-induced hyperlipidaemia models:

A) Animal

Wister albino male rats weighing 150–200 g will be acclimatised for 7

days to the laboratory at temperature (25 ± 1) °C, 12 h light–dark cycles, kept
in standard plastic cages of 5 rats each, given standard rat chow and water ad
libitum.
B) Induction of hyperlipidemia
Hyperlipidemia will be induced in each group of six experimental rats by
both single intraperitoneal injection of freshly prepared solution of Triton X-
100 (100 mg/kg b.w.) in physiological saline (0.9% NaCl solution) after
overnight fasting for 18 h and high-fat diet-induced hyperlipidaemia in rats.
C) Animal treatment
. The hyperlipidemia-induced rats will be treated with standard drug
atorvastatin, or with the polyherbal formulations developed from methanolic
and aqueous extracts of three medicinal plants i.e., cinnamon bark, arjuna
bark, fenugreek seeds and guggul resin at a single dose (300, 500, 700 and
1000 mg/kg b.wt)/day for 7 days orally.
D) Collection and treatment of blood samples
Twenty- four (24) hours after the last treatment, the rats will be sacrificed
and blood will be collected by incising the jugular vein into
Ethylenediaminetetraacetic acid (EDTA) and plain sample tubes for
haematological and serum analysis respectively.

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 E) Lipid profile and haematological analysis
Serum total cholesterol (TC), triglycerides (TG), low density lipoprotein
cholesterol (LDL) and high density lipoprotein cholesterol (HDLc) will be
estimated using the procedure outlined in commercial kits.
III. Submission of products and reports to the funding agency.

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 Annexure II

Who would be the beneficiary of this innovation and why?

The present innovative research work will be used for different aged groups especially
those who are suffering from hiperlipidaemia. As the purchase of statin drugs are highly
expensive for individuals of middle class and deprived sections of society, this polyherbal
formulation will be helpful or useful to the patients in the treatment of hyperlipidaemia.

At present the drugs used for hyperlipidemia are bile acid sequestrants, nicotinic acid,
fibric acid derivatives, and 3-hydroxy-3-methylglutaryl-coenzyme A (HMG Co-A) reductase
inhibitor i.e., Statins. Among these, statins and fibrates have shown greater promise.

New fibrates are known to be contraindicated in renal failure. Fibrates in combination

with Statins causes rhabdomyolysis. As these are causing side effects, drugs from plant sources
will play a vital role in replacing the drugs of synthetic origin. The main advantage of the drugs
obtained from plant origin doesn’t cause any side effects and they help maintain the health in
good condition without altering its condition.

So, the poly-herbal oral formulation going to be developed will fulfill the needs of
patients who are suffering from hyprlipidaemia and help reduce the risk of complications like
heart attack, stroke and ischemic organ disease including dementia which are developed due to
hyprlipidaemia.