Download formulation and evaluation of in-situ ophthalmic gel of

“FORMULATION AND EVALUATION OF IN-SITU OPHTHALMIC GEL OF
MOXIFLOXACIN AND KETOROLAC TROMETHAMINE COMBINATION”
MASTER OF PHARMACY DISSERTATION PROTOCOL
SUBMITTED TO THE
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA,
BANGALORE
BY
SOUDHA BEGUM
M.PHARM - I
Under the Guidance
Of
DR. HARISH.N.M.M.PHARM ,Ph.D
ASSOCIATE PROFESSOR
DEPARTMENT OF INDUSTRIAL PHARMACY
SRINIVAS COLLEGE OF PHARMACY,
VALACHIL, MANGALORE-574143.
2013-2014
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
BANGALORE,KARNATAKA
ANNEXURE - II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1.
Name of the Candidate and Address:
MISS. SOUDHA BEGUM
1st YEAR M.PHARM,
DEPARTMENT OF INDUSTRIAL PHARMACY
SRINIVAS COLLEGE OF PHARMACY,
VALACHIL, MANGALORE-574143.
2.
Name of the Institution:
SRINIVAS COLLEGE OF PHARMACY,
VALACHIL, FARANGIPETE POST,
MANGALORE-574143.
3.
Course of Study and Subject:
MASTER OF PHARMACY
(INDUSTRIAL PHARMACY)
4.
Date of Admission:
5.
Title of the Project:
25/07/2013
“FORMULATION AND EVALUATION OF IN-SITU OPHTHALMIC
GEL OF MOXIFLOXACIN AND KETOROLAC TROMETHAMINE
COMBINATION”
6.0
Brief resume of the intended work
6.1 Need of the study
Eye is the most vital organ of the body. To achieve effective ocular therapy, an adequate
amount of active ingredients must be delivered and maintain at the site of action within the eye. The
anatomical structure and the protective physiological process of the eye exert a formidable defense
against ophthalmic drug delivery, leads to poor precorneal drug loss results in poor ocular
bioavailability and ultimately poor ocular therapy. To improve ophthalmic drug bioavailability, there
are considerable efforts directed towards newer drug delivery systems for ophthalmic administration.
Since Conventional delivery systems often result in poor bioavailability and therapeutic response
because of high tear fluids turn over and dynamics cause rapid elimination of the drug from the eyes.
Newer research in ophthalmic drug delivery systems is directed towards a amalgamation of several
drug delivery technologies, that includes to build up systems which is not only extend the contact time
of the vehicle at the ocular surface, but which at the same time slow down the removal of the drug.
There are various new dosage forms like in-situ gel, collagen shield, minidisc, ocular film, ocusert,
nanosuspension, nanoparticulate system, liposomes, niosomes, dendrimers, ocular iontophoresis etc.
So, to overcome bioavailability problems, ophthalmic in-situ gels were developed. This deals with the
study of a novel in-situ gel approach as a means to localize and prolong drug activity at its site of
action. These are solutions, instilled as drops into the eye and undergo a sol to gel transition in the culde-sac, improved ocular bioavailability by increasing the duration of contact with corneal tissue,
thereby reducing the frequency of administration required in case of conventional ophthalmic
solutions, thus optimizing ocular therapy.1
Eye-drops are the conventional dosage forms that account for 90% of currently accessible
ophthalmic formulations. Despite the excellent acceptance by patients, one of the major problems
encountered is rapid precorneal drug loss. To improve ocular drug bioavailability, there is a significant
effort directed towards new drug delivery systems for ophthalmic administration. This chapter will
focus on three representative areas of ophthalmic drug delivery systems: polymeric gels, colloidal
systems, cyclodextrins and collagen shields. Hydrogels generally offer a moderate improvement of
ocular drug bioavailability with the disadvantage of blurring of vision. In-situ activated gel-forming
systems are preferred as they can be delivered in drop form with sustained release properties. Colloidal
systems including liposomes and nanoparticles have the convenience of a drop, which is able to
maintain drug activity at its site of action and is suitable for poorly water-soluble drugs. It seems that
new tendency of research in ophthalmic drug delivery systems is directed towards a combination of
several drug delivery technologies. There is a tendency to develop systems which not only prolong the
contact time of the vehicle at the ocular surface, but which at the same time slow down the elimination
of the drug. Combination of drug delivery systems could open a new directive for improving results
and the therapeutic response of non-efficacious systems. One of the ways to optimize ocular drug
delivery is to prolong the precorneal drug residence time. This focus on recent findings on the
formulation effects in ocular drug bioavailability, employing polymers for the preparation of
hydrogels, bioadhesive dosage forms, in-situ gelling systems and colloidal systems including
liposomes and nanoparticles. The results observed suggested that mucoadhesion or bioadhesion played
a role in the sustained action of drugs more significantly compared to non-mucoadhesive polymers.2
In-situ hydrogels are instilled as drops into the eye and undergoes a sol to gel transition in the cul-desac, improved ocular bioavailability by increasing the duration of contact with corneal tissue, thereby
reducing the frequency of administration.
Clinical applications of Moxifloxacin hydrochloride and Ketorolac tromethamine3:Moxifloxacin hydrochloride is long acting fourth generation fluoroquinolones having high
activity against gram-positive bacteria including beta lactam/macrolide resistant ones and some
anaerobes. It is used against acute bacterial exacerbations of chronic bronchitis, Community acquired
pneumonia, sinusitis, otitis media. It shows the bactericidal effect by specifically inhibiting the DNA
gyrase (topoisomerase II) and topoisomerase IV, which is required by bacteria for DNA replication,
transcription repair and recombination.
Ketorolac tromethamine is an novel NSAID with potent analgesic and modest antiinflammatory activity. In post-operative pain it has similar efficacy as opioids and is free of opioid side
effects. Like other NSAIDs, it inhibits prostaglandin synthesis and relives the pain by peripheral
mechanism. In short-lasting pain, it has compared favourably with aspirin.
The purpose of the present work is to develop an in-situ ophthalmic gel of Moxifloxacin
hydrochloride, a fluoroquinolone antibiotic and Ketorolac tromethamine combination for the treatment
of bacterial infection and reduction of ocular pain.
6.2 Review of Literature
1) Hareesh B, Mohammed G, Narayana R4 have developed and evaluate in-situ gels of
moxifloxacin for the treatment of periodontitis using gellan gum and sodium alginate based on
the concept of ion activated systems. The system utilizes polymers that exhibit sol-to-gel phase
transition due to change in specific physico-chemical parameters. Sol-to-gel transformation
occurred in the presence of monovalent /divalent cations. It was found that increase in the
concentration of calcium ions produced stronger gels. Formulations were evaluated for gelling
capacity, drug content, clarity, viscosity, gel strength, spread ability, microbiological studies
and in vitro release. The results shown by the above characterization studies were found to be
satisfactory. Experimental part showed that viscosity of sols and gel strength was increased
with increase in the concentration of polymers and the sustained release of drug was observed.
2) Sirish V, Sadhna K, Nalini S, Sadanandam M5 have formulated and evaluated ion activated
ocular gels of ketorolac tromethamine by using ion activated gelling systems of Ketorolac
tromethamine. These gelling systems involve the use of Gelrite as polymer. The formulations
were evaluated for clarity, pH measurement, gelling capacity, drug content estimation,
rheological study, in vitro drug release, ocular irritancy studies (as per draize test) and ex-vivo
corneal permeation studies using isolated goats cornea. The developed formulations showed
sustained release of drug for upto 6 hrs. The formulations were found to be non‐irritating with
no ocular damage.
3) Shashank N, Bharani S, Thakur R6 had designed and characterized pH triggered in-situ
ophthalmic gel forming solution using a fourth generation fluoroquinolone antibiotic,
Moxifloxacin hydrochloride. Polyacrylic acid (Carbopol 934) was used as gelling agent in
combination with hydroxyl propyl methyl cellulose(k15M) as viscosifying agent.
Benzalkonium chloride in suitable concentration was used as preservative. The formulations
were sterilized by autoclaving at 1210 C for 15 min at 21 psi. The prepared formulation were
evaluated for visual appearance, clarity, pH measurement, gelling capacity, drug content and
in-vitro diffusion studies, sterility test, microbiological studies and stability studies. Under
rheological condition both solution and gel have shown pseudo plastic behaviour.
4) Hanan L, Demiana N, Shoukry M7 had developed and evaluate temperature triggered in-situ
gelation using pluronic, and pH triggered in-situ gelation using carbopol 934 The developed
formulae were evaluated regarding their gelation temperature (for PL systems), gelling
capacity,
rheological
measurements.
characteristics,
in-vitro
release
behavior
and
mucoadhesion
5) Kavitha K, Rajas N8had prepared sustained release in-situ ocular gels of levofloxacin
hemihydrates using gelrite as gel forming polymer, which is used in treatment of various
bacterial infections. Formulations were evaluated for physical parameters like Clarity, pH,
drug content, gelation, rheological studies, sterility test, in-vitro drug release study and ocular
irritancy studies. The formulated gels were transparent, uniform in consistency and had
spreadability with a pH range of 7.1 to 7.4. Six different formulations with increasing polymer
concentrations were prepared which was found to have drug content of 72-86%. From the
preliminary studies it was observed that as the concentration of polymer was increased, the rate
of drug release decreased to produce sustained drug delivery for prolonged period of more than
8 hours. A maximum of90.2% drug release was observed in in-vitro studies. Further in-vivo
results conclude that it is be possible to formulate in-situ ocular gel containing Levofloxacin
Hemihydrate.
6) Saritha M, Amulya CH9 had developed ophthalmic drug delivery system using two different
gelling agents with different mechanisms for gelation of Moxifloxacin hydrochloride, a
fluoroquinolone antibiotic. Sodium alginate – an ion sensitive gelling agent and Poloxamer –
a temperature sensitive gelling agent were employed in the formation of in-situ hydrogel along
with HPMC K4M as viscosifying agent. The developed formulations were therapeutically
efficacious, stable, non-irritant and provided sustained release of the drug over an 8 hour.
7) Sumegha M, Surendrakumar J, Nishiprakash J10 had formulate and evaluate in-situ gelling
system of ketorolac tromethamine using Carbopol 940 as the gelling agent in combination with
HPMC K4M which acted as a viscosity enhancing agent. The prepared formulations were
evaluated for pH, clarity, gelling capacity, in-vitro drug release, drug-excipient interactions
(FTIR), and Short-term stability studies (40±2° C/75±5% RH for 3 months). IR spectroscopic
studies indicated that there are no drug-excipient interactions. drug release data of optimized
formulation was treated according to Zero, First, Korsmeyer Peppas and Higuchi kinetics to
access the mechanism of drug release. Eye irritation test using the HET-CAM test data of
optimized formulation and calculated S value. The gel provided sustained drug release over an
9 hour period and the reduced frequency of instillation resulting in better patient acceptance.
8) Basavaraj N, Manjappa S, Murthy R., Yuvaraj P11had formulate and evaluate an ophthalmic
delivery system for a nonsteroidal anti-inflammatory drug, ketorolac tromethamine, based on
the concept of pH-triggered in-situ gelation using Polyacrylic acid (Carbopol® 934) was used
as the gelling agent in combination with hydroxypropylmethylcellulose (Methocel K4M)
which acted as a viscosity enhancing agent. Compatibility studies of the drug excipients were
carried out using differential scanning calorimetry (DSC). The prepared formulations were
characterized for clarity, pH, drug content, sol-to-gel transition by scanning electron
microscopy (SEM), in-vitro and in-vivo drug release, ocular irritation and stability. The clarity,
pH and drug content of the developed formulation were found to be satisfactory. The
developed formulation was therapeutically efficacious, stable, non-irritant, and provided
sustained drug release over an 8-h period.
9) Hiremath et al12 had prepared and evaluated novel in-situ gum based ophthalmic drug delivery
system of linezolid. Hydroxypropyl guar (HPG) and xanthum (XG) were used as gum with the
combination of hydroxyethyl cellulose (HEC), carbopol (CP), and sodium alginate as viscosity
enhancing agents. Suitable concentrations of buffering agents were used to adjust the pH to
7.4. All the formulations were sterilized in an autoclave at 121°C for 15mins. The formulations
were evaluated for clarity, pH measurement, gelling capacity, drug content estimation,
rheological study, in-vitro diffusion study, antibacterial activity, isotonicity testing, eye
irritation testing. The developed formulations exhibited sustained release of drug from
formulation over a period of 6 hours thus increasing residence time of the drug.
6.3 Objectives of the study:The present research study is planed with the following objectives.
1) To formulate the in-situ ophthalmic gel of moxifloxacin hydrochloride and ketorolac
tromethamine combination.
2) To check the compatibility of moxifloxacin hydrochloride and ketorolac tromethamine with all
the polymers by using FTIR.
3) To evaluate the prepared formulation for12
a) Visual appearance
b) Clarity
c) pH measurement
d) Gelling capacity
e) Drug content
f) In-vitro diffusion studies
g) Sterility test
h) Microbiological studies
4) To carry out rheological studies.
5) To carry out stability studies as per ICH guidelines.
7
Material and Method
Materials:

Drug: Moxifloxacin hydrochloride and Ketorolac tromethamine.

Polymers: Gellan gum, sodium alginate,Gelrite, Carbomer, Cellulose acetate phthalate,
Poloxamers (Pluronic,Tetronics), cellulose derivatives (MC, HPMC), Xyloglucan
etc.

Excipients: Calcium chloride, Sodium citrate, Benzalkonium chloride etc.

Method13:
The ophthalmic gel of moxifloxacin hydrochloride and ketorolac tromethamine will be formulated by
using any one of the following method:

pH trigged method

Ion activated method

Temperature activated method
7.1 Source of Data:
Review of literature from

Journals such as,
o European Journal of Pharmaceutical Sciences.
o Journal of Drug Delivery and Therapeutics
o Indian Journal of Pharmaceutical sciences
o Asian Journal of Pharmaceutical Sciences
o World Journal of Pharmaceutical Sciences
o International Journal Pharmatech Research
o International Journal Of Pharmacy And Pharmaceutical Sciences
o International Journal Of Pharmaceutical Science Research And Review
7.2 – Method of Collection of Data:
1. An Overview of ophthalmic gel for sustained drug delivery.
2. Different approaches for achieving sustained drug delivery by using gel systems.
3. Different methods of preparing ophthalmic gel
4. Evaluation of various parameters of prepared gel formulations as follows: pH
 Drug Content
 In-vitro Drug Release
 Rheological Studies
 Stability Studies
 Microbiological studies
 Gelling capacity
 Ocular irritation studies
7.3-Does the study require any investigation to be conducted on patients or other humans
Or animals? If so, please describe briefly.
NOT APPLICABLE
7.4
Has ethical clearance been obtained from your institution in case of 7.3
NOT APPLICABLE
List of References:1. Lalit K, Ravindra S, Stuti G, Dhiraj K. In-situ Gel: A Novel System For Ocular Drug Delivery.
Int J Pharm Sci Rev Res 2011; 9(2): 83-91.
2. Sampath K, Debjit B, Shravan P, Shweta S. Recent Challenges and Advances in Ophthalmic
Drug Delivery System. The Pharm Inno 2012; 1(4): 19-31.
3. Tripathi KD, Quinolones, General consideration, Essential of Medical Pharmacology, 6th edn,
New Delhi:Jaypee Brothers Medical Publishers(P) Ltd; 2008: 687-693.
4. Hareesh B, Mohammed G, Narayana R. Development And Evaluation Of In- Situ Gels Of
Moxifloxacin For The Treatment Of Periodontitis. Indonesian J Pharm 2012; 23(3): 141 – 146.
5. Sirish V, Sadhna K, Nalini S, Sadanandam M. Formulation And Evaluation Of Ion Activated
Ocular Gels Of Ketorolac Tromethamine. Int J Curr Pharm Res 2010; 2(3): 33-38.
6. Shashank N, Bharani S, Thakur R. Formulation And Evaluation Of pH Triggered Ophthalmic
Gel Of Moxifloxacin Hydrochloride. Int J Pharmacy Pharm Sci 2012; 4(2): 452-459.
7. Hanan L, Demiana N, Shoukry M. Evaluation of two in-situ gelling systems for ocular delivery
of Moxifloxacin: In-vitro and in-vivo studies. J Chem Pharm Res 2011; 3(2): 66-79.
8. Kavitha K, Rajas N. Sustained Ophthalmic Delivery of Levofloxacin Hemihydrate from An Ion
Activated In-Situ Gelling System. Int J PharmTech Res 2011; 3(2): 702-706.
9. Saritha M, Amulya CH. Opthalmic Drug Delivery By Controlled Release In-Situ Forming
Moxifloxacin Hydrogel. Int J Adv Pharm Sci 2012; 3(5): 375-379.
10. Sumegha M, Surendrakumar J, Nishiprakash J. Formulation And Evaluation Of In-Situ
Opthalmic Gel Of Ketorolac Tromethamine. World J Pharmacy Pharm Sci 2013; 2(3): 13701384.
11. Basavaraj N, Manjappa S, Murthy R., Yuvaraj P. A novel pH-triggered in-situ gel for sustained
ophthalmic delivery of ketorolac tromethamine. Asian J Pharm Sci 2009; 4(3): 189-199.
12. Hiremath SP, Dasankoppa F, Nadaf A, Jamakandi VG, Jameel M, Sreenivas A, Hasanpasha S,
Aezazahmed, Nanjundaswamy G. Formulation and Evaluation of a Novel In-Situ Gum Based
Ophthalmic Drug Delivery System of Linezolid. Sci Pharm 2008; 76: 515–532.
13. Pandya T, Modasiya M, Patel V. Opthalmic in-situ gel sytems. IntJ Pharm Life Sci 2011; 2(5):
730-738.
9.
Signature of candidate
(SOUDHA BEGUM)
10.
Remark of the guide
The work, which is assigned to SOUDHA
BEGUM is under my guidance.
11
Name & Designation of
Dr. HARISH.N.M. M.PHARM, Ph.D
11.1 Guide
Associate Professor,
Department of Industrial Pharmacy
Srinivas College Of Pharmacy,
Valachil, Mangalore-574143.
11.2 Signature
11.3 Co-Guide (if any)
11.4 Signature
11.5 Head of the Department
Dr. HARISH.N.M. M.PHARM, Ph.D
Associate Professor
Department of Industrial Pharmacy
Srinivas College of Pharmacy,
Valachil, Mangalore-574143.
11.6 Signature
12
12.1 Remark of the Director principal
Recommended
and
forwarded
for
favourable
consideration
12.2 Signature
Dr. A. R. SHABARAYA M.PHARM, Ph.D
Principal and Director,
Srinivas College Of Pharmacy,Mangalore-574143.