Levetiracetam is an anti-epileptic medicine used to treat seizures in epilepsy. It is used for partial-onset, myoclonic, or tonic–clonic seizures. General route of administration is by Oral and IV. But it is not recommended throughout an epileptic attack due to the risk of nausea or vomiting. The Intranasal (IN) route is a promising therapy option, as it allows medications to reach the brain directly, it is regarded an alternative to parenteral administration, and hence the effective dosage predicted via other administration routes is scheduled to be reduced using the IN route

1. Presented by
RAKTIMAVA DAS SARKAR
M.Pharm(Pharmaceutics) – 2nd Semester
Roll- 19320322015
BENGAL SCHOOL OF TECHNOLOGY,
SUGANDHA, HOOGHLY -712102

2. CONTENTS
CONTENTS
1. Introduction
2. Review of Literature
3. Materials
4. Methods
5. Results
6. Discussion
7. Conclusion
8. Reference

3. INTRODUCTION
1) To formulate surfactant-based nanovesicles of Levetiracetam
2) For use as intranasal delivery as a brain-targeted Anti-Epileptic Delivery System
3) To prepare Nanovesicles via Solvent Evaporation Technique.
4) To Optimize formulations by the Design-Expert® program
Levetiracetam is an anti-epileptic medicine used to treat seizures in epilepsy. It is used for partial-
onset, myoclonic, or tonic–clonic seizures. General route of administration is by Oral and IV. But it
is not recommended throughout an epileptic attack due to the risk of nausea or vomiting. The
Intranasal (IN) route is a promising therapy option, as it allows medications to reach the brain
directly, it is regarded an alternative to parenteral administration, and hence the effective dosage
predicted via other administration routes is scheduled to be reduced using the IN route.
Different pathways have been suggested for IN route: the olfactory and trigeminal nerve and/or the
systemic path.
Fig.1: Structure of Levetiracetam

4. REVIEW OF LITERATURE
Ren et al. has concluded in his review article that vesicles are particularly important drug vehicles
for oral delivery of various therapeutical by virtue of structural diversity, great flexibility for drug
loading, and prominent ability in absorption promotion.
Liu et al. summarised that, purified and characterized nanovesicles from Artemisia annua revealed
its therapeutic potential through a specific pathway.
Ventosa et al. developed a new type of stable pH-sensitive nanovesicles, based on quantosomes, for
delivery of miRNA.
Bezabeh et al. prepared Amphotericin B and Miltefosine nanovesicles by the ethanol injection
method. The study showed that entrapment efficiency was significantly affected by the effects of
lipids, drug-lipid proportion, ethanol-water composition and stirring rate.

5. MATERIALS AND METHODS
MATERIALS PROCURED FROM USED AS
Levetiracetam El Andalous Medical
company (Cairo, Egypt)
API for treatment of
Seizures
Commercial
Levetiracetam
(Tiratam®)
El Andalous Medical
company (Cairo, Egypt)
Commercially Available
Dosage Form
Sorbitan
tristearate
(Span® 65)
Sigma-Aldrich Company
(St. Louis, MO, USA).
Oil Soluble Surfactant for
Stability
Triethanolamine
98%
Sigma-Aldrich Company
(St. Louis, MO, USA).
pH Adjuster
Ethanol 95% El-Nasr Pharmaceutical
chemicals Co. (Cairo,
Egypt)
Solvent
Acetonitrile El-Nasr Pharmaceutical
chemicals Co. (Cairo,
Egypt)
Solvent
Chloroform El-Nasr Pharmaceutical
chemicals Co. (Cairo,
Egypt)
Solvent
MATERIALS PROCURED
FROM
USED AS
Sodium
hydroxide
El-Nasr Pharmaceutical
chemicals Co. (Cairo,
Egypt)
Buffer Component
Potassium
Dihydrogen
Phosphate
El-Nasr Pharmaceutical
chemicals Co. (Cairo,
Egypt)
Buffer Component
Cholesterol Loba Chemie (Mumbai,
India)
Vesicular Layer
Component
Carbopol 940 Athos chemicals
(Gujarat, India)
Gel Forming Agent
VISKING ®
Dialysis Tubing
molecular weight
cutoff 12,000–
14,000 pore
diameter 25 A °
SERVA Electrophoresis
(Uetersen, Germany).
Dialyser
Table 1: Chemicals used in Formulation.

6. MATERIALS AND METHODS
Accurately weighed amount of Span
65 and Cholesterol were dissolved in
Chloroform
Accurate amount of
Drug(Levtiracetam) was
dissolved in ethanol
The aq. Phase was added to the
organic phase and stirred
continuously 900RPM
After Completion of evaporation of
organic phase the formed
nanovesicles were cooled.
Organic
Phase
Aqueous
Phase
Mixture
Solvent
Evaporation
Fig.2: Solvent Evaporation Technique for Nanovesicle Preparation
Preparation of Nanovesicles

7. MATERIALS AND METHODS
Independent Variables
Levels
-1 0 +1
X1: Span 65 (mg) 25 50 75
X2: Cholesterol (mg) 25 50 75
Dependent Variables Constraints
Y1: Particle size (nm) Minimum
Y2: Entrapment efficiency
(%)
Maximum
Y3: Zeta potential (mV) Maximum
Formulations Drug(mg) Span 65(mg) CHL(mg)
F1 50 25 25
F2 50 50 25
F3 50 75 25
F4 50 25 50
F5 50 50 50
F6 50 75 50
F7 50 25 75
F8 50 50 75
F9 50 75 75
F(Opt.)
Using Design-Expert®
program version 10.0
50 42.954 62.419
Table 2: Independent Variables.
Table 3: Dependent Variables.
Table 4: Formulation Design.

8. MATERIALS AND METHODS
Gel code Carbopol 940
concentration
(%W/V)
Nanovesicle
content (%W/V)
G 1 1% 45.5%
G 0.75 0.75% 45.5%
G 0.375 0.375% 45.5%
Preparation of Levetiracetam Surfactant-
based Nanovesicles Gel
The specified amounts of Carbopol 940
were dispersed gradually in deionized
water using a magnetic stirrer.
The stirring was maintained at 100RPM
after the gel base had formed.
Disperse in each formula 45.5% W/V of the
selected optimized Levetiracetam surfactant-
based nano-vesicles.
followed by adjusting the pH to
approximately 6.0 using triethanolamine
Table 5: Gel Formualtion Design.

9. MATERIALS AND METHODS
Determination of percentage drug
Entrapment Efficiency (%EE)
Particle size (PS) analysis, polydispersity index
(PDI), and zeta (ζ)-potential
1 milliliter of the nano vesicular dispersion
subjected for cooling ultracentrifuged at
20,000 RPM for 30 min at 4 ᵒC
The precipitated nanovesicles were washed with
1 ml phosphate buffer (pH 6.8)
Re-centrifuged for an additional 10 min to ensure
the extraction of the unentrapped drug
EE% = [(Total amount of drug − amount of free drug)
/Total amount of drug] x 100
Nanovesicles (Lev-Nvs) dispersion was
established using dynamic light scattering
By using Malvern Zeta sizer at 25 °C at a
scattering angle of 90°
Optimization of Formulation
Design-Expert® program version 10.0, an optimized
formula F(OPT) was conducted.
This formula was further used to
prepare and evaluate the gel.

10. MATERIALS AND METHODS
Animal handling and drug injection Chromatographic conditions
Nine healthy male Wistar rats weighing between
250 and 300 g were employed.
Three groups of rats were randomly allocated. All rats
were fasted for 12 h and given access to water on an ad
libitum basis.
Group I underwent the commercial Tiratam® solution.
Group II underwent the developed nasal gel, 50-µl nasal
gel into each nostril.
The negative control group was used to provide blank
plasma.
Levetiracetam was measured utilizing an assessed
HPLC(Agilent 1200 series) for determination in plasma.
Chromatography was performed on a C18 column.
MP is 50 mmol KH2PO4 buffer and
acetonitrile(9:1v/v). NaOH (1 ml/min) was used to
adjust the pH of the mobile phase.
Statistical Analysis
The pharmacokinetic features, AUC(0–t), AUC(0-∞),
Cmax, and Tmax, were examined statistically via analysis
of variance (ANOVA). The P-value was measured from
the gathered F-value utilizing IBM SPSS statistics version
20 Microsoft software
Quantification was performed using UV detection at
205 nm.

11. With the aid of Design-Expert® program version 10.0, an optimized formula F(OPT) was conducted.
RESULTS
Fig.3: TEM images of optimized levetiracetam surfactant-based
nanovesicles formulation.
Form. E. E.% Particle size
(nm)
PDI Zeta
potential
(mV)
F1 81.31±2.3 219.13±30.03 1 -32.1
F2 83.19±2.0
3
266.03±22.56 1 -40.5
F3 65.19±2.3 137.26±15.50 1 -28
F4 66.46±0.5 362.16±10.68 0.695 -19.9
F5 88.4±0.53 372.46±93.41 0.73 -24.9
F6 67.35±0.5
6
381.46±20.16 0.818 -24.3
F7 74.42±0.5
2
373.83±143.2 0.759 -28.6
F8 76.02±0.9
7
329±39.5 0.789 -28.5
F9 77.35±0.5
6
516.2±65.03 0.532 -25.1
F(Opt.) 87.9±1.06 206.7±20.43 0.979 -34.1
PK parameters Commercial Oral
Solution
Nasal Gel
𝐶𝑚𝑎𝑥(ng/ml) 10.421±0.295 21.26±1.615
AUC(h*ng/ml) 23.135±0.43 58.715±7.74
𝑇𝑚𝑎𝑥(h) 0.917±0.144 0.5±0
𝑇1/2(h) 2.18±0.714 3.467±0.601
MRT(h) 2.186±0.025 2.88±0.17
𝐾𝑒(1/h) 0.345±0.124 0.204±0.037
Table 7. PK Parameters Prepared using PKSolver,
Table 6: Physical Parameters.

12. FTIR of the drug, crabapol, span 65, cholesterol, and physical mixture
RESULTS
Fig.4: FTIR of the drug, carbapol, span 65, cholesterol, and
physical mixture
Fig.6: Levetiracetam plasma conc.-time profile for oral
solution and nasal gel. All data showed as mean±SD (n=3)
Fig.5: In vitro release of levetiracetam drug from solution, levetiracetam
nanovesicle formulation and (1, 0.75 and 0.375%) Carbopol nanovesicle
gels.
Fig.7: Illustration of size and zeta potential of levetiracetam surfactant-based
nanovesicles (Lev-Nvs) formulation

13. FTIR of the drug, crabapol, span 65, cholesterol, and physical mixture
DISCUSSION
ϴ The F(opt) was the best formulation amongst all the 10 formulations, by means of Entrapment Efficiency,
Particle Size, Zeta Potential and Porodispersity Index.
ϴ Figure 6. of the Plasma Drug Concentration vs Time graph, it is clearly visible that the Nano-vesicular
Nasal Gel Formulation has a better plot in terms of various Pharmacokinetic parameters.
ϴ Solvent Evaporation technique proved to be a suitable technique for preparing Nanovesicles.
ϴ Figure 4. represented that Levetiracetam FT-IR spectra have shown carbonyl peak at 1680 cm1, NH
stretching absorption 3500–3200 cm-1, 2989 cm-1 CH stretching, CN stretching at 1083 cm-1. FT-IR Study
confirmed that there are no physical interactions among the excipients.
ϴ TEM study shows that the Nanovesicles were evenly formed as a spherical structure which eases drug
delivery.

14.  FTIR of the drug, crabapol, span 65, cholesterol, and physical mixture
CONCLUSION
 Levetiracetam was successfully incorporated into (cholesterol/span 65) nanovesicle which is intended
for intra-nasal delivery.
 Using Design-Expert®, 𝐹𝑂𝑃𝑇 was selected as the best formulation with an
1. entrapment efficiency of 87.9±1.06%,
2. 206.7±20.43 nm particle size,
3. -34.1mV zeta potential and
4. 0.979 Porodispersity Index.
 Further preparation of nanovesicle nasal gels. G (0.375) nasal gel formulation indicated a favourable
(87.36 %) drug release in a span of 12 h.
 In vivo release study showed a prolonged release of the nanovesicle nasal gel and a higher relative
bioavailability (293.85%) when contrasted to a similar dose of oral solution.
 Thus, nanovesicle nasal gels are superior to oral solutions for antiepileptic drug administration and
patient compliance.

15.  FTIR of the drug, crabapol, span 65, cholesterol, and physical mixture
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