FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH

RAMIPRIL AND ALPRAZOLAM”

Index table

Formulation Evaluation and characterization of the niosomes

Loaded with ramipril and alprazolam

A SYNOPSIS SUBMITTED TO INSTITUTE OF PHARMACY, BUNDELKHAND UNIVERSITY IN THE

PARTIAL FULFILLMENT FOR THE AWARD OF THE

DEGREE OF

MASTER OF PHARMACY

IN

PHARMACEUTICS

SESSION: 2019- 2020

UNDER THE SUPERVISION OF

GUIDE SUBMITTED BY

Dr. PEEYUSH BHARDWAJ HIMANSHU CHAUHAN

(Associate Professor) (B.PHARMA)

Institute of pharmacy ROLL. NO: 141251016222

Bundelkhand University, En. No.: BU/14/10213

Jhansi

INSTITUTE OF PHARMACY BUNDELKHAND UNIVERSITY, JHANSI (U.P.)

1. INTRODUCTION
A Niosome is a non-ionic surfactant-based Vesicle (biology and chemistry). Niosomes are
formed mostly by non-ionic surfactant and cholesterol incorporation as an excipient. Other
excipients can also be used. Niosomes have more penetrating capability than the previous
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
RAMIPRIL AND ALPRAZOLAM”
preparations of emulsions. They are structurally similar to liposomes in having a bilayer,
however, the materials used to prepare niosomes make them more stable. It can entrap both
hydrophilic and lipophilic drugs, either in an aqueous layer or in a vesicular membrane made of
lipid material.
Material used in the preparation of noisome

At first the surfactant cholesterol which is about 150 micro ml is poured in the

Rotary evaporator which contain 10 ml of diethyl ether (10) ml

 Then it is evaporated in the rotary evaporator

 After the evaporation in the rotary evaporator hydration is done
 Hydration cause the surface to swell up
 Then it folds to form vesicle

PROPERTIES OF NOISOME

Based on the vesicle size

Niosomes can be divided into three groups

These are small unilamellar vesicles (SUV, size=0.025-0.05 μm),

Multilamellar vesicles (MLV, size=>0.05 μm),

Large Unilamellar vesicles (LUV, size=>0.10 μm).

Methods of Preparation
Niosomes are prepared by different methods based on the sizes of the vesicles and their
distribution, number of double layers, entrapment efficiency of the aqueous phase and
permeability of vesicle membrane.

Preparation of small unilamellar vesicles

SONICATION
The aqueous phase containing drug is added to the mixture of surfactant and cholesterol in a
scintillation vial. The mixture is homogenized using a sonic probe at 60°C for 3 minutes. The
vesicles are small and uniform in size.
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RAMIPRIL AND ALPRAZOLAM”

Micro fluidization
Two fluidized streams move forward through precisely defined micro channel and interact at
ultra-high velocities within the interaction chamber. Here, a common gateway is arranged such
that the energy supplied to the system remains within the area of niosomes formation. The result
is a greater uniformity, smaller size and better reproducibility.

Preparation of multilamellar vesicles

Hand shaking method (Thin film hydration technique)
In the hand shaking method, surfactant and cholesterol are dissolved in a volatile organic solvent
such as diethyl ether, chloroform or methanol in a rotary evaporator, leaving a thin layer of solid
mixture deposited on the wall of the flask. The dried layer is hydrated with aqueous phase
containing drug at normal temperature with gentle agitation.

Trans-membrane pH gradient (inside acidic) drug uptake process (remote Loading)

Surfactant and cholesterol are dissolved in chloroform.The solvent is then evaporated under
reduced pressure to obtain a thin film on the wall of the round-bottom flask. The film is hydrated
with 300 mM citric acid (pH 4.0) by vortex mixing. The multilamellar vesicles are frozen and
thawed three times and later sonicated. To this niosomal suspension, aqueous solution containing
10 mg/ml of drug is added and vortexed. The pH of the sample is then raised to 7.0-7.2 with 1M
disodium phosphate. This mixture is later heated at 60°C for 10 minutes to produce the desired
multilamellar vesicles.

Preparation of large unilamellar vesicles

Reverse phase evaporation technique (REV)
In this method, cholesterol and surfactant are dissolved in a mixture of ether and chloroform. An
aqueous phase containing drug is added to this and the resulting two phases are sonicated at 4-
5°C. The clear gel formed is further sonicated after the addition of a small amount of phosphate
buffered saline. The organic phase is removed at 40°C under low pressure. The resulting viscous
niosome suspension is diluted with phosphate-buffered saline and heated in a water bath at 60°C
for 10 min to yield niosomes.

Ether injection method

The ether injection method is essentially based on slow injection of niosomal ingredients in ether
through a 14-gauge needle at the rate of approximately 0.25 ml/min into a preheated aqueous
phase maintained at 60°C. The probable reason behind the formation of larger unilamellar
vesicles is that the slow vaporization of solvent results in an ether gradient extending towards the
interface of aqueous-nonaqueous interface. The former may be responsible for the formation of
the bilayer structure. The disadvantages of this method are that a small amount of ether is
frequently present in the vesicles suspension and is difficult to remove.
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
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Miscellaneous
Multiple membrane extrusion method
A mixture of surfactant, cholesterol, and diacetyl phosphate in chloroform is made into thin film
by evaporation.The film is hydrated with aqueous drug solution and the resultant suspension
extruded through polycarbonate membranes, which are placed in a series for up to eight
passages. This is a good method for controlling niosome size.

Niosome preparation using polyoxyethylene alkyl ether

The size and number of bilayer of vesicles consisting of polyoxyethylene alkyl ether and
cholesterol can be changed using an alternative method. Temperature rise above 60°C transforms
small unilamellar vesicles to large multilamellar vesicles (>1 μm), while vigorous shaking at
room temperature shows the opposite effect, ie, transformation of multilamellar vesicles into
unilamellar ones. The transformation from unilamellar to multilamellar vesicles at higher
temperature might be the characteristic for polyoxyethylene alkyl ether (ester) surfactant, since it
is known that polyethylene glycol (PEG) and water remix at higher temperature due to
breakdown of hydrogen bonds between water and PEG moieties. Generally, free drug is removed
from the encapsulated drug by gel permeation chromatography dialysis method or centrifugation
method. Often, density differences between niosomes and the external phase are smaller than
that of liposomes, which make separation by centrifugation very difficult. Addition of protamine
to the vesicle suspension facilitates separation during centrifugation.

Emulsion method
The oil in water (o/w) emulsion is prepared from an organic solution of surfactant, cholesterol,
and an aqueous solution of the drug. The organic solvent is then evaporated, leaving niosomes
dispersed in the aqueous phase.

Lipid injection method

This method does not require expensive organic phase. Here, the mixture of lipids and surfactant
is first melted and then injected into a highly agitated heated aqueous phase containing dissolved
drug. Here, the drug can be dissolved in molten lipid and the mixture will be injected into
agitated, heated aqueous phase containing surfactant.

Niosome preparation using Micelle

Niosomes may also be formed from a mixed micellar solution by the use of enzymes. A mixed
micellar solution of C16 G2, dicalcium hydrogen phosphate, polyoxyethylene cholesteryl
sebacetate diester (PCSD) converts to a niosome dispersion when incubated with esterases.
PCSD is cleaved by the esterases to yield polyoxyethylene, sebacic acid and cholesterol.
Cholesterol in combination with C16 G2 and DCP then yields C16 G2 niosomes

Advantage of noisome preparation

FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
RAMIPRIL AND ALPRAZOLAM”
• Niosomes are osmotically active, chemically stable and have long storage
time compared to liposome’s

• Their surface formation and modification is very easy because of the

functional groups on their hydrophilic heads

• They have high compatibility with biological systems and low toxicity
because of their non-ionic nature

• They are biodegradable and non-immunogenic

• They can entrap lipophilic drugs into vesicular bilayer membranes and
hydrophilic drugs in aqueous compartments

• They can improve the therapeutic performance of the drug molecules by

protecting the drug from biological environment, resulting in better
availability and controlled drug delivery by restricting the drug effects to
target cells in targeted carriers and delaying clearance from the circulation
in sustaied drug delivery

• Access to raw materials is convenient

RATIONALE FOR COMBINATION THERAPY

When hypertensive patients do not achieve adequate control of their blood pressure, the
options to try and achieve required treatment goals are to increase the dose of monotherapy or
to use drug combinations with minimum side effects. In order to avoid complications, it is
important to start treatment as soon as possible, achieve the goals in the shortest time possible
and ensure treatment adherence.
The mechanisms that lead to a blood pressure increase in a patient are diverse – monotherapy
acts on one or at best two of these mechanisms, while the use of combinations of drugs allows
for action on several different hypertensive mechanisms. By combining two drugs with different
mechanisms of action, an antihypertensive effect of two to five times greater than that
obtained by monotherapy is possible. Increasing the dose of monotherapy reduces coronary
events by 29% and cerebrovascular events by 40%, while combining two antihypertensive
agents with a different mechanism of action reduces coronary events by 40% and
cerebrovascular events by 54%. Thus, the use of combination therapy provides greater
protection to a target organ than increasing the dose of monotherapy.
In summary, combination therapy looks like a better option than increasing the dose.
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
RAMIPRIL AND ALPRAZOLAM”
BLOOD PRESSURE A BRIEF INTRODUCTION
Many psychotherapeutic and CNS-active agents exhibit hypotensive effects, especially during
initiation of therapy and dose escalation. Co-administration with anti-hypertensives and other
hypotensive agents, in particular vasodilators and alpha-blockers, may result in additive effects
on blood pressure and orthostasis.

ABOUT BLOOD PRESSURE

Blood pressure is the force of your blood pushing against the walls of your arteries. Each time
your heart beats, it pumps blood into the arteries. Your blood pressure is highest when your
heart beats, pumping the blood. This is called systolic pressure. When your heart is at rest,
between beats, your blood pressure falls. This is called diastolic pressure.Your blood pressure
reading uses these two numbers. Usually the systolic number comes before or above the
diastolic number. For example, 120/80 means a systolic of 120 and a diastolic of 80. High blood
pressure usually has no symptoms. So the only way to find out if you have it is to get regular blood
pressure checks from your health care provider. Your provider will use a gauge, a stethoscope or
electronic sensor, and a blood pressure cuff. He or she will take two or more readings at separate
appointments before making a diagnosis.

ANTI- HYPERTENSIVE MEDICINE

On giving medicine as the combined therapy the blood pressure may have positive effect on the
body, and therefore the alprazolam and ramapril combination is Important to give a patience as
they have positive effect on the blood pressure regulation.

ACE inhibitors may cause bone marrow suppression, rarely in uncomplicated individuals but
more frequently in patients with renal impairment, especially if they also have a collagen-
vascular disease such as systemic lupus erythematosus or scleroderma. Neutropenia,
agranulocytosis, aplastic anemia, hemolytic anemia, eosinophilia and thrombocytopenia have
been reported, mostly with captopril. Therapy with ACE inhibitors should be administered
cautiously in patients with pre-existing blood dyscrasias or complications that may increase the
risk of bone marrow depression during ACE inhibitor therapy. Monitoring of blood counts,
particularly white blood cells, should be considered.

ANTI-HYPERTENSIVE MEDICATION
DIURETICS

Diuretics help the kidneys eliminate excess salt and water from the body's tissues
and blood.
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RAMIPRIL AND ALPRAZOLAM”
CALCIUM CHANNEL BLOCKERS
Calcium channel blockers block the entry of calcium into muscle cells in artery
walls. And thus help in the maintenance of the blood pressure and this is one of
the effective way of medication which helps in the blood pressure regulation.

ACE INHIBITORS
ACE inhibitors inhibit the activity of angiotensin-converting enzyme (ACE), an enzyme
responsible for the conversion of angiotensin I into angiotensin, a potent vasoconstrictor.

ADRENERGIC RECEPTOR ANTAGONISTS

Beta blockers are used for the maintaining of the blood pressure and they play an important
role in the management of that blood pressure
VASODILATOR
Vasodilators act directly on the smooth muscle of arteries to relax their walls so blood can
move more easily through them; they are only used in hypertensive emergencies or when other
drugs have failed, and even so are rarely given alone.

Sodium nitroprusside, a very potent, short-acting vasodilator, is most commonly used for the
quick, temporary reduction of blood pressure in emergencies (such as malignant
hypertension or aortic dissection). Hydralazine and its derivatives are also used in the
treatment of severe hypertension, although they should be avoided in emergencies. They are
no longer indicated as first-line therapy for high blood pressure due to side effects and safety
concerns, but hydralazine remains a drug of choice in gestational hypertension.

RENIN INHIBITORS
Renin comes one level higher than angiotensin converting enzyme (ACE) in the renin–
angiotensin system. Inhibitors of renin can therefore effectively reduce
hypertension. Aliskiren (developed by Novartis) is a renin inhibitor which has been approved by
the U.S. FDA for the treatment of hypertension

CURRENTLY AVAILABLE MEDICINE FOR TREATMENT OF BLOOD PRESSURE

The treatment of HTN in 2014 predominantly involved older medications in 5 major classes of
drugs: ACEIs, thiazide diuretics, ARBs, CCBs, and beta-blockers. Selection of antihypertensive
agents showed limited variation by age, gender, race, and insurance type. Although 58% of
treated patients had SBP <140, 24% had poorly controlled HTN with SBP ≥150, indicating the
need for improved treatment.
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
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SELECTION OF DRUG
Hear we have selected the drug for the purpose to treat hypertension, and combination
therapy would be better for the treatment of the hypertension and therefore the two of drug
are Ramipril and alprazolm

ABOUT RAMIPRIL

Ramipril is used to treat high blood pressure (hypertension). Lowering high blood pressure helps prevent
strokes, heart attacks, and kidney problems. Ramipril is also used to improve survival after a heart
attack. It may also be used in high risk patients (such as patients with heart disease/diabetes) to help
prevent heart attacks and strokes. Ramipril may also be used to treat heart failure in patients who have
had a recent heart attack.

Ramipril is an ACE inhibitor and works by relaxing blood vessels so that blood can flow more
easily.Ramipril is available under the following different brand names: Altace. drug

Drug Profile

Generic name Altace

Molecular formula C23H32N2O5
Molecular wt. 416.5g/mol
Structure

Chemical name (2S,3aS,6aS)-1[(S)-N [(S)-1-Carboxy-3-phenylpropyl]

alanyl]
IUPAC name (2S,3aS,6aS)-1-[(2S)-2-[[(2S)-1-ethoxy-1-oxo-4-
phenylbutan-2-yl]amino]propanoyl]-3,3a,4,5,6,6a-
hexahydro-2H-cyclopenta[b]pyrrole-2-carboxylic acid
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
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Dosage form Tablet

Potency 1.5 mg, 2.5 mg,
Melting point
Solubility Soluble in water
Half life 2 to 4 hour
Indications Ramipril (Altace) is an ACE inhibitor. ACE stands for
angiotensin converting enzyme.

Ramipril is used to treat high blood pressure

(hypertension) or congestive heart failure, and to
improve survival after a heart attack.

Ramipril may also be used for purposes not listed in

this medication guide.

Mechanism of action Ramipril inhibits the RAAS system by binding to and

inhibiting ACE thereby preventing the conversion of
angiotensin I to angiotensin II. As plasma levels of
angiotensin II fall, less activation of the G-protein coupled
receptors angiotensin receptor I (AT1R) and angiotensin
receptor II (AT2R) occurs
Absorption 50% to 60% is absorbed after oral administration.
Protein binding 73 %

Drug profile

ABOUT ALPRAZOLAM

Alprazolam is used to treat anxiety and panic disorders. It belongs to a class

of medications called benzodiazepines which act on the brain and nerves (central nervous
system) to produce a calming effect. It works by enhancing the effects of a certain natural
chemical in the body (GABA).
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
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Take this medication by mouth as directed by your doctor. Dosage is based on your medical
condition, age, and response to treatment. Your dose may be gradually increased until the
drug starts working well. Follow your doctor's instructions closely to reduce the risk of side
effects.

If you suddenly stop using this medication, you may have withdrawal symptoms (such
as seizures). To help prevent withdrawal, your doctor may lower your dose slowly.
Withdrawal is more likely if you have used alprazolam for a long time or in high doses. Tell
your doctor or pharmacist right away if you have withdrawal.

Though it helps many people, this medication may sometimes cause addiction. This risk may
be higher if you have a substance use disorder (such as overuse of or addiction to
drugs/alcohol). Take this medication exactly as prescribed to lower the risk of addiction. Ask
your doctor or pharmacist for more details.

When this medication is used for a long time, it may not work as well. Talk with your doctor if this
medication stops working well.

Drug profile
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
RAMIPRIL AND ALPRAZOLAM”

Sr. No Systematic (IUPAC) name

1 8-chloro-1-methyl-6-phenyl-4H-[1,2,4]triazolo[4,3-a][1,4]benzodiazepine

Sr.No Clinical Data

1 Trade name Xanax, Xanax

2 Other name Niravam.

3 AHFS/Drug.com
4 Medline plus

Sr.no PHARMACOKINETIC DATA

1 Bioavailability 84-91% in 1.8 hr

2 Protein binding 80 %
3 Metabolism metabolism of alprazolam is
mediated largely through the
action of CYP3As
4 Half life The half life is 16.3h in the elderly,
5.8-65.3h in patients with
alcoholic liver disease, 9.9-40.4h in
obese patients
5 Excretion

Sr. No CHEMICAL DATA

1 Formula C17H13CIN4
FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
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2 Molecular mass 308.8 g/mol

3 Density

MECHANISM OF ACTION
Alprazolam is classed as a high-potency triazolobenzodiazepine a benzodiazepine with
a triazole ring attached to its structure. As a benzodiazepine, alprazolam produces a
variety of therapeutic and adverse effects by binding to the GABA benzodiazepine
receptor site and modulating its function; GABA receptors are the most prolific inhibitory
receptor within the brain. The GABA chemical and receptor system mediates inhibitory
or calming effects of alprazolam on the nervous system. Binding of alprazolam to the
GABAA receptor, a chloride ion channel enhances the effects of GABA, a
neurotransmitter. When GABA binds the GABA receptor the channel opens and
chloride enters the cell which makes it more resistant to depolarization. Therefore,
alprazolam has a depressant effect on synaptic transmission to reduce anxiety.
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Step 1 GABA RECEPTOR MODULATOR

Step 2 ALPRAZOLAM

Step 3 INCREASES GABAAR Open Frequency

Step 4 Neuronal Hyper polarization

Step 5 Decrease CNS Excitability

Step 6 Hepatic Metabolism (CYP3A4 substrate)

Step 7 ½- life: 11-16h Renal Excretion

FORMULATION, EVALUATION AND CHARACTERIZATION OF THE NIOSOMES LOADED WITH
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Alprazolam side effects

Serious side effects

Mental problems. Symptoms can include:

o Depressed mood
o Thoughts of suicide
o Confusion
o Hallucinations (seeing or hearing things that aren’t real)
 Movement problems. Symptoms can include:
o Uncontrolled muscle movements
o Tremor
o Seizures
 Heart problems. Symptoms can include:
o Chest pain
o Abnormal heartbeat
 Liver problems. Symptoms can include:
o Jaundice (with yellowing of the skin or the whites of the eyes)
 Urinating less than usual or not at all

Alprazolam may interact with other medications

Alprazolam oral tablet can interact with other medications, vitamins, or herbs
you may be taking. An interaction is when a substance changes the way a
drug works. This can be harmful or prevent the drug from working well.
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Drugs that should not be used with alprazolam

Do not take these drugs with Xanax. When used with Xanax, these drugs
can cause dangerous effects in the body. Examples of these drugs include:

 Certain types of antifungals, such as itraconazole or ketoconazole.

When used with Xanax, these drugs can cause increased drowsiness.

Drugs that increase the risk of side effects

Taking alprazolam with certain medications raises your risk of side effects from those
drugs. If you take one of these drugs with alprazolam, you may have increased
drowsiness.

 Sedatives or hypnotics,
 Anxiolytics,
 Narcotic analgesics,
 Sedative antihistamines,
 Anesthetics,
 Antidepressants,
 Antacids,

 Oral contraceptives (birth control pills). If you take one of these drugs with
alprazolam, you may have increased drowsiness or other side effects.

Alcohol interaction

Alprazolam can cause drowsiness, dizziness, or lightheadedness. The use of drinks that contain
alcohol raises your risk of these side effects. Try to avoid drinking alcohol while you take this
drug.
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Warnings for people with certain health conditions

For people with acute narrow-angle glaucoma : This drug can make your
condition worse. Do not take this drug if you have acute narrow-angle
glaucoma.

For people with a history of alcoholism, drug abuse, or personality disorder: This
drug can lead to physical and psychological dependence (addiction). If you have a
history of these conditions, you have a greater risk of dependence on thi s drug.

For people with liver disease: It may be harder for your body to break down this drug.
This can increase the amount of the drug in your body, which can lead to more side
effects.

For people with obesity: It may be harder for your body to break down this drug. This
can increase the amount of the drug in your body, which can lead to more side effects.

For people with severe lung disease: This drug can make your condition worse. Talk
with your doctor about whether this drug is safe for you.