DESIGN AND DEVELOPMENT OF SELF EMULSIFYING DRUG … · Self emulsifying drug delivery systems (SEDDS) are isotropic mixtures of natural or synthetic oils, solids or liquid surfactants

www.wjpr.net Vol 8, Issue 9, 2019. 263

Rajendra et al. World Journal of Pharmaceutical Research

DESIGN AND DEVELOPMENT OF SELF EMULSIFYING DRUG

DELIVERY SYSTEM OF MARAVIROC

Patel Rajendra*, Patel Dr. Arun, Patel Shailendra, Dwivedi Neelesh and Patel Bhavesh

Department of Pharmaceutics, Shri Ram Group of Institutions Faculty of Pharmacy, Near Iti

Madhotal, Jabalpur. (M.P.).

ABSTRACT

Self emulsifying drug delivery systems (SEDDS) are isotropic

mixtures of natural or synthetic oils, solids or liquid surfactants or

alternatively, one or more hydrophilic solvents, and co-solvents or co-

surfactants Maraviroc SEDDS formulation when diluted with 250

times gives globule size of 110 nm and zeta potential of -5.18. There is

no significant difference in globule size when diluted with medias like

0.1 N HCl and Phosphate buffer of pH 6.8). Maraviroc drug release

from the SEDDS formulation was performed in 0.1 N HCl and 87.92

(± 4.78) % released within 15 minutes. In-vitro drug diffusion study in

dialysis membrane showed that 80.56 ± 2.81% maraviroc diffused

from the SEDDS formulation in the Phosphate buffer of pH 7.4 in five

hours, whereas only 42.86 (±2.12) % released from Maraviroc

suspension at the same conditions. Maraviroc SEDDS formulation can be used in the

treatment of AIDS.

KEYWORDS: AIDS, MARAVIROC, Self emulsifying drug delivery systems, HIV.

1 INTRODUCTION

Acquired immune deficiency syndrome (AIDS) is characterized by progressive loss of the

CD4C helper/inducer subset of T lymphocytes. Loss of T cells leads to severe impairment of

immune function, constitutional diseases, opportunistic infections, neurological

complications (AIDS dementia complex), and neoplasms that seldom occur in persons with

intact immune function.[1-10]

While the precise mechanisms that result in destruction of the

immune system are not completely understood, an abundance of epidemiologic, virologic,

World Journal of Pharmaceutical Research SJIF Impact Factor 8.074

Volume 8, Issue 9, 263-275. Review Article ISSN 2277– 7105

Article Received on

25 May 2019,

Revised on 16 June 2019,

Accepted on 06 July 2019

DOI: 10.20959/wjpr20199-15398

*Corresponding Author

Patel Rajendra

Department of

Pharmaceutics, Shri Ram

Group of Institutions

Faculty of Pharmacy, Near

Iti Madhotal, Jabalpur.

(M.P.).



and immunologic data support the conclusion that infection with HIV (human

immunodeficiency virus) is the underlying cause of AIDS.

HIV was originally designated human T lymphotropic virus (HTLV)-III, lymphadenopathy-

associated virus (LAV), or AIDS-associated retrovirus (ARV). AIDS is induced by the HIV

virus. Therefore, it is specifically referred to as HIV/AIDS because other factors such as

corticosteroids, cancer chemotherapy, and alkylating agents can also produce AIDS-like

symptoms

1.1 Structure of HIV

HIV is a lentivirus of the family Retroviridae, mostly known for being the causative agent of

AIDS.[13]

This virus can be seen as a biological nanostructure (around 100–150 nm). Its outer

envelope or coat is composed of a double layer of lipid envelope that bears numerous spikes.

Each spike is composed of four molecules of gp120 and the same number of gp41 embedded

in the membrane. Beneath the envelope is a layer of matrix protein that surrounds the core

(capsid). The capsid has a hollow, truncated cone shape and is composed of another protein,

p24 that contains the genetic material of the HIV virus. Two strands of RNA consisting of

about 9200 nucleotide bases, integrase, a protease, ribonuclease, and two other proteins, p6

and p7, fit inside the viral core.

1.2 HIV pathogenesis

After initial infection and local amplification at the mucosal site, infected cells migrate to

regional lymph nodes, leading to a mild initial viral amplification in naïve T cells (39). The

viral infection is then quickly disseminated by T cells to lymphoid organs, particularly the

gut-associated lymphoid tissues (GALT), spleen, and bone marrow, being accompanied by a

burst in the viral load (acute infection).[40]

During the acute and early stages of infection the

gastrointestinal tract is particularly affected by the virus, leading to a dramatic loss of CD4+

and CD8+ T cells which never quite recover completely and remain despite antiretroviral

treatment.[41-42]

Individuals during this acute phase pose an increased risk for sexual

transmission as result of high blood and genital viral load, with clear implications in the

prevention of HIV transmission.[43]

CD4+ cell levels recover soon after; in the case of CD8+

cells there is a rise followed by a rapid recovery of normal levels. Levels of the virus are then

down regulated in response to the intense immune response, but never completely depleted,

resulting in clinical latent and asymptomatic infection.[44]

During latency, the virus persists

particularly in extra-vascular tissues, lymph node dendritic cells and resting CD4+ memory



cells. This state can evolve to a symptomatic clinical stage (usually several years after the

initial infection), designated by AIDS, which is characterized by decreased CD4+ T cell

counts and rising viral load.[13]

With the progression of infection, HIV genetic diversity

increases noticeably due to intense error-prone reverse transcription and evolutionary

pressure to evade the immune system.[36]

This new heterogeneous population dramatically

increases the generation of viruses resistant to cellular and humoral immune response, and

represents a major challenge in the development of therapy and preventative strategies.

1.3 General principles of HIV therapy

Antiretroviral therapy (ART) of HIV-1 was initiated in 1987 with the use of zidovudine

(AZT). Although it was initially hoped that HIV- 1 could be eradicated from the body, it is

now clear that virus suppression is the best that can be achieved by continuous ART. Therapy

can fail due to low adherence or the evolution of drug-resistant HIV-1 variants, while drug-

toxicity may be another reason to stop therapy. HAART aims to prevent emergence of drug-

resistant HIV-1 variants by suppressing its replication to negligible levels. Also, it is very

difficult for the virus to acquire complex combination of mutations required for multi-drug

resistance. In cases when the virus succeeds, it usually pays great fitness costs and becomes

less aggressive within the given host and less transmissible to other hosts. A major barrier to

curing HIV infection remains the ability of HIV to integrate in the host genome and remain

latent. HAART cannot completely eradicate the virus from the organism and patients have to

commit to the therapy for life.

1.4 Limitations of available therapies

Even if HAART regimens present considerable anti-HIV activity, several factors frequently

compromise its success. To begin with, current therapy is not able to provide a cure mainly

because of HIV's ability to persist in latency state in cellular and anatomical reservoir sites.

Beside this fact, problems of current antiretroviral therapy also include prolonged treatment

periods with drugs possessing important adverse effects, poor drug-regimen compliance, drug

resistance, drug–drug interactions, poor drug pharmacokinetics, viral levels rebound after

therapy cessation, and costs.

2. SELF EMULSIFYING DRUG DELIVERY SYSTEMS

Self emulsifying drug delivery systems (SEDDS) are defined as isotropic mixtures of natural

or synthetic oils, solids or liquid surfactants or alternatively, one or more hydrophilic

solvents, and co-solvents or co-surfactants.[53]

SEDDS are able to self emulsify rapidly into



fine O/W emulsion in the gastrointestinal fluids, under gentle agitation provided by the

gastrointestinal tract. This fine O/W emulsion results in small droplets of oil dispersed in the

gastrointestinal fluids that provide a large interfacial area enhancing the activity and

minimizing the irritation due to contact of drug in the gut wall. Self Emulsifying System

(SES) can be formulated with little energy input and the shelf life is longer than conventional

emulsions (54, 55-58). SEDDS typically produce emulsion with a droplet size between 100 to

300nm, while self micro emulsifying drug delivery system (SMEDDS) form transparent

micro-emulsions with a drop let size of less than 50nm.

2.1 Advantages of SEDDS

1. SEDDS is novel approach to improve water solubility and ultimate bioavailability of

lipophilic drugs.

2. Many drugs show large inter-subject and intra-subject variation in absorption leading to

variable plasma profile. SEDDS produce reproducible plasma profile.

3. Fine oil droplets empty rapidly from the stomach and promote wide distribution of the

drug throughout the intestinal tract thus minimizing the irritation frequently encountered

with extended contact of drug and gut wall.

4. As compared to emulsions, which are sensitive and metastable dispersed forms, SEDDS

are physically stable formulations that are easy to manufacture.

5. SEDDS require very simple and economical manufacturing facility like simple mixture

with agitator and volumetric liquid filling equipment for large-scale manufacturing.

6. SEDDS/SMEDDS has potential to deliver peptides that are prone to enzymatic hydrolysis

in GastroI intestinal tract (GIT).[59]

3.1 PREFORMULATION STUDIES

3.1.1. Physico-chemical Characterization of Drug

Organoleptic evaluation revealed that the drug is a white crystalline powder.

3.2. Analytical method development

3.2.1. Development of spectrophotometric method for the detection of Maraviroc

Experimentally working wavelength (λ) in different medium such as methanol, 0.1N HCl,

Phosphate buffer pH 7.4 was determined & found to be 210 nm. The standard calibration

curves for Maraviroc in above mentioned medias with regression value (R2) are shown in

figure 1. The coefficient of correlation showed excellent correlation between concentration

and absorbance with the coefficient of variance (CV) in the range of 0.999 to 1. The relation



between drug concentration and absorbance is linear and the curves obeys Beer-Lambert’s

law within the concentration range of 2-12 µg/ml of Maraviroc. Calibration curve of

methanol, 0.1N HCl, and Phosphate buffer pH 7.4 was used for solubility analysis of drug in

various oils & surfactants in-vitro dissolution studies and in-vitro permeation study

respectively.

Table 1: Concentration and absorbance of standard drug solutions in different media.

Sr. No Concentration

(µg/ml)

Absorbance

Methanol 0.1 N HCl PB pH 7.4

1 2 0.037 0.034 0.029

2 4 0.075 0.064 0.058

3 6 0.108 0.097 0.089

4 8 0.145 0.130 0.119

5 10 0.180 0.158 0.148

6 12 0.212 0.191 0.178

Figure 1: Regressed calibration curve of Maraviroc in methanol.

Figure 2: Regressed calibration curve of Maraviroc in 0.1N HCl.



Figure 3: Regressed calibration curve of Maraviroc in Phosphate buffer pH 7.4.

Formulation of self emulsifying drug delivery system (SEDDS)

3.3.1 Solubility Studies of Maraviroc in various oils and surfactants

Table 2: - Solubility of Maraviroc in various oils.

SN Oils Solubility of Maraviroc (mg/mL)

1 Peceol 92±4.2

2 Miasine 35-1 85±3.6

3 Labrafil 1944 CS 62±5.3

4 Labrafil 2125 CS 65±4.8

5 Soya bean oil 40±3.1

6 Olive oil 38±3.9

Figure 4: Solubility of Maraviroc in various oils.

Table 3: Solubility of Maraviroc in various Surfactants and co-surfactants.

SN Surfactant/

Co-surfactant Solubility mg/mL

1 Transcutol HP 185±6.86

2 Labrasol 78±3.64

3 Span 80 60±2.9

4 Tween 80 42±3.7

5 Cremophor EL 34±4.1

6 PEG 400 46±3.2



Figure 5: Solubility of Maraviroc in various Surfactants.

Solubility Studies of Maraviroc in various ratios of surfactant & co-surfactant

Table 4:- Solubility Study of Maraviroc in various ratio of Transcutol HP: Labrasol.

Sr. No. Transcutol :Labrasol Amount in (mg/ml)

1 1:1.1 128.53

2 1:1.2 116.74

3 1:1.3 104.43

3.3.3 Development of pseudo ternary phase diagram

Three separate pseudo ternary phase diagrams (Figure 6) were prepared using water and

Peceol. Third phase was selected from either of Labrasol, transcutol HP and combination of

Labrasol and Transcutol HP., The gray area in the figure (Figure 7) indicates clear

microemulsion region in the system. As shown in the figures, the area meant for the

composition of o/w microemulsion increased with increasing surfactants/co-surfactant ratio.

The result confirms that the amount of surfactants, co-surfactants play an important role for

formulation of self emulsifying system for the selected oil.

Figure 6: Pseudo ternary phase diagram.



4. Preparation of liquid SEDDS

Oils, surfactant, surfactant/co-surfactant ratio were selected on the basis of Maraviroc

solubility, Table 11 shows list of ingredients with their chemical name & solubility selected

for the optimized liquid SEDDS formulation. Table 5 provides optimized formula of liquid

SEDDS.

Table 5: Oil surfactant selected for preparation of self emulsifying system.

Oils/Surfactant (Chemical name) Solubility* (mg /ml)

Peceol (Glyceryl monooleate) 92.60±4.2

Transcutol HP (Diethylene glycol monoethyl ether) 185.37±6.86

Labrasol (Caprylocaproyl macrogol-8 glycerides) 78.44±3.64 * Mean ±SD (n=3)

Table 6: Optimized formula of liquid self emulsifying system (SES).

Sr.No Ingredients Amount (mg) Quantity (%)

1 Maraviroc 100 10.16

2 Peceol 95 9.64

3 Labrasol 450 45.68

4 Transcutol HP 340 34.52

5 Total 985 100

5. Evaluation of SEDDS formulation

5.1.1 Particle size measurement

Self emulsifying system was diluted up to 250 times individually with distilled water, 0.1 N

HCl, and Phosphate buffer pH 6.8. The droplet size of the resultant emulsion was determined

by Zeta sizer (nano ZS90, Malvern, UK). The mean droplet size and polydispersity index of

the liquid SEDDS are shown in table 7. Emulsion droplet size distribution curve is shown in

figure 8.

5.1.2 Zeta potential

Optimized SEDDS was diluted up to 250 times with distilled water. Zeta potential of the

diluted emulsion was determined by Zeta sizer (nano ZS90, Malvern, UK).

Zeta potential of the optimized formulation was found to be -5.18.



Figure 7:- Zeta potential in water.

5.1.3. Dilution Potential

The prepared formulation was diluted 250 times with continuous media (distilled water, 0.1

N HCl, Phosphate buffer pH 6.8). There was no significant difference in the globule size of

diluted SEDDS formulation after dilution by different medium at the beginning and after 8

hour of dilution.

Table 7: Globule size of diluted SEDDS formulation.

Dilution medium Distilled water 0.1 N HCl Phosphate buffer pH 6.8

Size (nm)

0 hour 108.5 110.7 112.1

8 hour 120.8 119.0 124.7

Figure 8:- Zeta potential in water and phosphate buffer pH 6.8.



5.1.4. In-vitro drug release study

In-vitro drug release of the SEDDS formulations was carried out using USP type I dissolution

apparatus (100 rpm; 37 ºC ± 0.5 ºC) in 0.1N HCl. 53.34 (± 3.12) % drug was released within

5 min and 87.92 (± 4.78) % in 15 min After 15 min, drug release rate became very slow and

rest amount of drug released within 60 minutes.

Table 8: Cumulative percentage drug release (n = 3).

SN Time (minutes) Cumulative % drug release

1 0 0

2 5 53.34±3.12

3 15 84.92±3.48

4 30 90.13±3.78

5 45 92.49±3.98

6 60 94.12±4.12

Figure 9: Cumulative percentage drug release of Maraviroc SEDDS.

In-vitro drug diffusion study

In-vitro diffusion study was performed using dialysis membrane to compare permeation of

drug between developed Maraviroc SEDDS and Maraviroc suspension through the dialysis

membrane. Comparative drug release profile is shown in figure. Nearly 45.54 ± 2.03% of

drug was released from Maraviroc SEDDS at very first hour against only 16.12 (±1.13) %

drug release from Maraviroc suspension at the same time. At the end of five hour period,

almost all the drug (80.56 ± 2.81%) diffused from the SEDDS formulation, whereas 42.86

(±2.12) % only released at the same time period from Maraviroc suspension.



Table 9: Cumulative percent drug diffused (n=3).

SN Time

(minute)

Cumulative % drug diffused

from Maraviroc SEDDS

Cumulative % drug diffused

from Maraviroc suspension

1 0 0 0

2 15 14.34±1.23 5.1±0.27

3 30 26.78±1.67 9.8±0.76

4 60 45.54±2.03 16.12±1.13

5 120 61.12±2.32 24.69±1.12

6 180 70.23±2.65 30.75±1.45

7 240 76.42±2.78 36.81±1.86

8 300 80.56±2.81 42.86±2.12

Figure 10: Cumulative percent drug diffused.

Accelerated stability studies

Developed SEDDS formulation was exposed to accelerated stability condition (400

C &75%

RH) to confirm its stability. The study was carried out for 2 months and the results are given

in Table 16.

Table 10: Evaluation of SEDDS exposed to accelerated stability condition.

Storage condition 40 ºC and 75% RH

SEDDS evaluation Sampling Schedule

Initial First month Second month

Color Clear Clear Clear

Globule size (nm) 108.5 114.1 115.5

Zeta potential (mV) -5.18 -5.36 -5.95

Assay (%) 99.067±1.11 98.5± 0.33 98.28±0.35

The stability testing results showed that the formulation was stable upto 2 months without

any change in physical property of the SEDDS formulation, including its color, globule size,

zeta potential, assay of drug.



CONCLUSION

Analytical method of Maraviroc by UV Visible spectrophotometer was developed at

wavelength () 210nm. The developed UV method was used for determination of Maraviroc

solubility in different excipients, in-vitro drug release and in-vitro drug diffusion study of

formulation. Oils, surfactants and co-surfactants were selected based on solubility of drug and

triglycerides of longer carbon chain length along with surfactant and co-surfactants with the

self emulsification potential. Peceol (solubility 92±4.2 mg/mL) was selected as oil while

Labrasol (solubility 78±3.64 mg/mL) and Transcutol HP (solubility 185±6.86 mg/mL) as

surfactant and co-surfactant respectively. Pseudo-ternary phase diagram was developed for

different ratios of oils, surfactants co-surfactants and Smix for microemulsion area. The

different ratios of excipients for microemulsion region in phase diagram were determined to

draw conclusion for self emulsifying system formulation

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