Formulation and in Vitroevaluation of Microspheres of Metoprolol Succinate

FORMULATION AND IN VITROEVALUATION OF MICROSPHERES OF METOPROLOL SUCCINATE

M.Pharm dissertation protocol submitted to

Rajiv Gandhi University of Health Sciences, Karnataka

Bangalore – 560041

By

Mr. DINESH KUMAR .A B.Pharm

Under the Guidance of

Dr. DIVAKAR GOLI M.Pharm, Ph.D

Professor Principal

Dept.of Pharmaceutics.

Department of Pharmaceutics

Acharya & B.M.Reddy College of Pharmacy

Soladevanahalli,Chikkabanavara (Post)

Hesarghatta main road, Bangalore – 560 090

2011– 2013

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE.

ANNNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1 / Name and address of candidate / Mr. Dinesh Kumar.A
D.no: 4-3-75/3,
Bhuktapur,
Near Anand bhavan,
Adilabad -533104
Adilabad Dt.,
ANDHRA PRADESH.
2 / Name of institution / ACHARYA & B.M. REDDY COLLEGE OF
PHARMACY.
Soladevanahalli, Hesarghatta Main Road,
Chikkabanavara Post.
Bangalore-560090
3 / Course of study and subject / M. Pharm
(Pharmaceutics)
4 / Date of admission /

29-12-2011

5 / Title of the project / FORMULATION AND IN VITRO EVALUATION OF MICROSPHERES OFMETOPROLOL SUCCINATE
6
6.1 / BRIEF RESUME OF INTENDED WORK
NEED FOR THE STUDY:
Hypertension or high blood pressure is a cardiac chronic medical condition in which the systemic arterial blood pressure is elevated. It is the opposite of hypotension. Hypertension is classified as either primary (essential) hypertension or secondary hypertension; About 90–95% of cases are categorized as "primary hypertension," which means high blood pressure with no obvious medical cause. The remaining 5–10% of cases (Secondary hypertension) is caused by other conditions that affect the kidneys, arteries, heart or endocrine system.
Persistent hypertension is one of the risk factors for stroke, myocardial infarction, heart failure and arterial aneurysm, and is a leading cause of chronic kidney failure1, 2, 3.
Anti-hypertensive are class of drugs which are used to manage hypertension. There are various classes of drugs which are used as antihypertensive like beta blockers, calcium channel blockers; ACE inhibitors etc., among all these, beta blockers are still being used as first line agents used for management of hypertension4.
Metoprolol succinate is a beta1-selective (cardio selective) adrenergic receptor blocking agent. This preferential effect is not absolute, however, at higher plasma concentrations, Metoprolol succinate also inhibits beta2-adrenoreceptors, chiefly located in the bronchial and vascular musculature. Metoprolol succinate has no intrinsic sympathomimetic activity and membrane-stabilizing activity is detectable only at plasma concentrations much greater than required for beta-blockade. Because of these desired pharmacodynamic properties, Metoprolol succinate is used popularly for management of hypertension.
Metoprolol succinate belongs to class I category in BCS classification system, i.e. freely soluble & highly permeable. Because of good solubility and permeability, its bioavailability and half life is less. This results in multiple doses of Metoprolol succinateevery day. Hence, continuous efforts are being made whereby number of doses of Metoprolol succinate can be minimized. This has resulted in formulation of Metoprolol succinate in to extended release form. Literature is flooded with reference in which various approaches are used for the formulation of extended release form of Metoprolol. These methods suffer from one or the other disadvantages. Hence, in the present work an attempt will be made in which formulation of Metoprolol succinate will be designed for extended release and it will be validated2.
Microsphereis small spherical particles, with diameters in themicrometerrange (typically 1μm to 1000μm (1mm)). Microspheres are sometimes referred to as microparticles4.
Microspheres are a class of multiple-unit dosage forms and offer the advantage of combining various types of subunits in a single system and offer much diversity in achieving dissolution profiles5.
The advantagesof oral controlled-release products over traditional systems are as follows: decreased fluctuation of serum concentrations resulting in reduced toxicity and sustained efficacy; and decreased frequency of dosing resulting in improved patient compliance, reduced patient care time, and possibly reduced total amount of drug used6.
In present study solvent evaporation method is used so its brief information is given.Solvent evaporation method is the most popular technique of preparing micro particles. The solvent evaporation technique was fully developed at the end of the 1970s. The technique of emulsion solvent evaporation offers several advantages; it is preferable to other preparation methods such as spray drying, sonication and homogenization, etc, because it requires only mild conditions such as ambient temperature and constant stirring. Thus, a stable emulsion can be formed without compromising the activity of the drug7.
6.2
6.3 / REVIEW OF LITERATURE:-
Literature survey was carried out on the proposed topic with the facility of internet and helinet and referring scientific journal. The survey reveals that, no work has undertaken on the proposed topic and some related research work are coated below.
Patro MN et al., prepared microspheres of 5- fluorouracil for colon targeting, usingEudragit S 100 and L 100 were used to prepare microspheres by a simple oil /water emulsification process. Process parameters were analyzed in order to optimize the drug loading and release profiles. In further attempts mixtures with Eudragit S100 and L100 were prepared to prolong drug release. Eudragit S100, pure or in mixture, was found to retain drug release at pH 4.5 lower than 41% within 6 h. At pH 7.4, nearly immediate release (within 30 min) was observed for pure S100, while mixtures enabled to prolong the release slightly. However, the formulation proved its applicability in-vitro as a promising device for pH-dependent colon delivery of 5-fluorouracil. The study was aimed at develop the porous microspheres, which can control the drug release up to 6 h8.
Lakshmana PS et al.,Developed sustained release microspheres of rosin containing aceclofenac, using rosin by o/w emulsion solvent evaporation technique. The effect of three formulation variables including the drug: polymer ratio, emulsifier (polyvinyl alcohol) concentration and organic solvent (dichloromethane) volume were examined. The prepared batches were characterized for microspheres particle size distribution, encapsulation efficiency and in vitro release behaviour. The study reveals that drug: polymer ratio had a considerable effect on the entrapment efficiency. Drug, polymer concentrations were varied to obtain optimum release profile for sustaining the action of thedrug9.
Dortunc B et al.,and co other workers Formulatedsolvent evaporation method using acetone/liquid paraffin system. The influence of formulation factors (stirring speed, polymer: drug ratio, type of polymer, ratio of the combination of polymers) on particle size, encapsulation efficiency and in vitro release characteristics of the microspheres were investigated. The yields of preparation and the encapsulation efficiencies were high for all formulations the microspheres were obtained. Although acetazolamide release rates from Eudragit RS microspheres were very slow and incomplete for all formulations, they were fast from Eudragit RL microspheres. When Eudragit RS was added to Eudragit RL microsphere formulations, release rates slowed down and achieved the release profile suitable for per oral administration10.
Sundar RS et al.,and co other researchers prepared Carprofen microspheres by emulsion solvent evaporation technique using cellulose acetate phthalate as the polymer. The prepared microspheres were performed in vitro drug release studies. The surface morphology and uniform coating of microspheres were characterized by scanning electron microscope. The drug content and drug entrapping efficiency of microspheres were also determined. The dissolution data registered a release of 88.39 % which complies with Indian pharmacopoeia official requirement11.
Barhate SD et al., developed prepared floating drug delivery system of ketorolactrometamolby emulsion solventdiffusion method by using ethyl cellulose, HPMC K4M, Eudragit R 100, Eudragit S 100 polymers in varying concentration. Formulations were evaluated for percent yield, particle size, entrapment efficiency, in vitro buoyancy and in vitro release studies. The optimized formulation shows good buoyancy and invitro controlled release of ketorolac trometamol12.
ChinnaGB et al.,formulatedmicrospheres by solvent evaporation method using an acetone / liquid paraffinsystem and Phase separation co-acervation method using petroleum ether and coconut oil as dispersion and continuous phase systems. Magnesium stearate was used as the droplet stabilizer and n-hexane was added to harden the microspheres. The prepared microspheres were evaluated for their micromeritic properties, drug content and encapsulation efficiency and characterized by Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). The in vitro release studies was performed by buffer change method to mimic Gas Intestine Tract(GIT) environment in pH 1.2, carbonate buffer (acidic) and pH 7.4, phosphate buffer
(Alkaline) 13.
Bhagwat DA et al., prepared microspheres of isosorbidedinitrate using the non-aqueous emulsification solvent evaporation method. The impacts of different factors such as stirring rate, concentration of acrycoat S 100 as matrix polymer on the characteristics of the microspheres were investigated. The morphology of microspheres was studied using optical and scanning electron microscopy and it was shown that microspheres had a spherical shape and smooth surface. The particle size of microspheres analyzed by optical microscopic method was affected by stirring rate and concentrations of acrycoat S 100. As the stirring rate increased the particle size decreases and as concentration of acrycoat increase the particle size also increases. Larger microspheres showed greater drug loading and smaller microspheres showed a faster drug release14.
Najmuddin Met al.,and co other workersprepared floating microspheres of ketoprofen using Eudragit S 100 and Eudragit L 100 as polymer. Floating microspheres of ketoprofen were prepared by emulsion solvent diffusion method using Eudragit S 100 and Eudragit L 100 as polymer. The floatingmicrospheres was evaluated such as micromeritic properties, particle size, percentage yield, in vitro buoyancy, incorporation efficiency, drug polymer compatibility (IR study ), scanning electron microscopy and drug release of microspheres. Results show that as increase in drug:polymer ratio affects the particle size, percentage yield, in vitro buoyancy and drug release of microspheres15.
Belgamwar V et al.,developedoral mucoadhesivemultiparticulate system containing metoprololtartarate: An In Vitro – Ex Vivo Characterization. Prepare mucoadhesivemultiparticulate system for oral drug delivery using ionic gelation technique. Microspheres composed of various mucoadhesive polymers including HPMC of various grades like K4M, K15M, K100M, E50LV, Carbopol of grades 971P, 974P and polycarbophil were prepared. In this technique cross linking of sodium alginate with calcium chloride was done which retarded the release of drug from the mucoadhesive polymer. The preliminary mucoadhesive strength studies performedfor various polymers using rotating cylindrical method showed that HPMC had greater mucoadhesive properties than carbopol and polycarbophil16.
Patrick BO et al.,and co workers formulated microspheres by the solvent evaporation technique using preparation of microspheres and microcapsules based on biodegradable polymers and copolymers of hydroxyl acids. The properties of biodegradable microspheres of poly(lactic acid) (PLA) and poly (lactic- glycolic acid) (PLGA) have been extensively investigated. The encapsulation of highly water soluble compounds including proteins and peptides presents formidable challenges to the researcher. The successful encapsulation of such entities requires high drug loading in the microspheres, prevention of protein degradation by the encapsulation method, and predictable release of the drug compound from the microspheres17.
 Philip PM et al., preparednelfinavirmesylate microcapsules, using cellulose acetate were developed as sustained release dosage form and release kinetics were studied.The desired microencapsulation was achieved by solvent evaporation method using cellulose acetate, performed by size, shape, entrapment efficiency, infrared spectroscopy and in vitro drug release analysis. The prepared microcapsules were free flowing, spherical in shape, with particle size in the range 80‐1000μm. FCA‐5 had maximum entrapment efficiency of 95.03%. The in vitro release profile of FCA‐5 was found to give 81.68% release of the drug which was more than the release of drug in FCA‐1, FCA‐2, FCA‐3 and FCA‐418.
Patel J et al., co workers developedpropanolol hydrochloride-loaded carbopol-934P/ethyl cellulose mucoadhesive microspheres,containing carbopol-934P as mucoadhesive polymer and ethyl cellulose as carrier polymer, were prepared by an emulsion-solvent evaporation technique. Results of preliminary trials indicated that the quantity of emulsifying agent, time for stirring, drug-to-polymers ratio, and speed of rotation affected various characteristics of microspheres. In-vivo evaluation studies on propranolol hydrochloride mucoadhesive microspheres and propranolol hydrochloride powder were performed on normal healthy rabbits. The prolonged gastrointestinal residence time and slow release of propranolol hydrochloride resulting from the mucoadhesive microspheres, could contribute to the provision of a sustained anti-hypertensive effect19.
OBJECTIVE OF THE STUDY:
The objectives of the present study are following:-
To carry out authentication and pre-formulation studies of drug metoprolol succinate.
To carry out drugs and polymer interaction studies.
Formulationofmicrospheres of metoprolol succinate.
Characterization of microspheres for various parameters..

• Particle size analysis.
• Percentage Yield.
• Drug content analysis.
• Percentage Entrapment efficiency.
• Scanning electron microscopy (SEM).
• Differential Scanning Calorimetry (DSC).

To carry out in vitro dissolution profile for microspheres.
To carry out stability studies on the most satisfactory formulation as per ICH guidelines.
7.0
7.1
7.2 / MATERIALS AND METHODS:
SOURCE OF DATA
1)Review of literature from:
a. Journals – such as

• Indian Journal of Pharmaceutical Sciences

• European Journal of Pharmaceutical Sciences
• International Journal of Pharmaceutics
• European journal of pharmaceutics and biopharmaceutics.
• Journal of controlled release.etc b .Reference books- such as

• Essentials of medical pharmacology, 6th edition- K D Thripathi
• Novel drug delivery systems, volume 50- Yie W. Chien etc

1. World Wide Web
2. J-Gate@Helinet, etc

MATERIALS
DRUG: Metoprolol succinate.
POLYMERS:Eudragit RS-100, Eudragit-RL 100.
EXCIPIENTS: excipients as required.etc
Method :
Formulation and evaluation of microspheres containing Antihypertensive drug by solvent evaporation method or any suitable /Developed method
METHOD OF COLLECTION OF DATA:

1. Authentication of drug:

a)Solubility studies.
b)Melting point determination.
c)Purity.
2. To carry out drugs and polymer interaction studies FT-IR studies.
3. Preparation of microsphere of Antihypertensive drug by solvent evaporation method or any
suitable /developed method.
4. Evaluation of the various properties of Microspheres.

• Percentage Yield
• Particle size analysis
• Percentage Entrapment efficiency
• Scanning electron microscopy (SEM)
• Differential Scanning Calorimetry (DSC)
• Drug content analysis.

To carry out in vitro release studies using U.S.P. dissolution test apparatus
Stability studies on the optimized formulation as per ICH guidelines at 30 ± 2°C (65 ± 5 %RH) and 40 ± 2°C (75 ± 5 %RH).
7.3

7.4 / DOES THE STUDY REQUIRE ANY INVESTIGATION TO BE CONDUCTED ON PATIENT OR OTHER HUMANS OR ANIMALS?
“ NO”
Has ethical clearance been obtained from your institution in case of 7.3?
“ not APPLIcable”
8.0 / LIST OF REFERENCES:-
1. Carretero OA, Oparil S. Essential hypertension. Part I: definition and etiology. circulation 2000; 101(3):329–35.
2.Hypertension [online]. [Cited on: 2012 May 21];
3. PierdomenicoSD, Nicola DM, Esposito AL. Prognostic value of different indices of blood pressure variability in hypertensive patients Am J Hypertens 2009; 22(8):842–7.
4. Microspheres [online]. [Cited on: 2012 May 21];
5. Lee T W, Robinson JR editor. Remington: The science and practice of pharmacy. 20th ed. Baltimore: Pharmaceutical Press; 2000.
6. Gupta PK, Robinson JR editor. Marcel Dekker: oral controlled-release delivery. Treatise on controlled drug delivery. New York: CPL Press; 1992.
7. Aftabrouchad D, Doelker E. Preparation methods for biodegradable microparticles loaded with water-soluble drugs. STP Pharma Sci 1992; 2(5):365-80.
8. Patro MN, Dhawale SC, Bankar AS. Formulation and evaluation porous microspheres of 5- fluorouracil for colon targeting. Int J PharmTech Res 2010;2(2):1112-8.
9.Lakshmana PS,Shirwaikar AA, Shirwaikar A, Kumar A. Formulation and evaluation of sustained release microspheres of rosin containing aceclofenac. Ars Pharm 2009; 50(2):51-62.
10. Dortunc B, Haznedar S. Preparation and in vitro evaluation of eudragit microspheres
containing acetazolamide.Int J Pharm 2004; 269: 131– 40.
11. Sundar RS, Murugesan A, Venkatesan P, Manavalan R. Formulation development and evaluation of carprofen microspheres. Int J PharmTech Res 2010; 2(3):1674-6.
12. Barhate SD, Rupnar YS,Sonvane RM,Pawar KR, Rahane RD. Formulation and evaluation of floating microspheres of ketorolac trometamol.Int J Pharm Res Dev 2009;1(9):1-8.
13.Chinna GB, Shyam SR,VarmaKM, Sleeva RM, Kiran SM. Formulation and evaluation of indomethacin microspheres using naturaland synthetic polymers as controlled release dosage forms. Int J Drug Discovery 2010; 2(1):8-16.
14. Bhagwat DA, Bhutkar MA, Todkar SS, Mohite SK, Gattani YS. Formulation and evaluation of controlled release microspheres of isosorbide dinitrate. Int J PharmTech Res 2009; 1(2):125-8.
15. Najmuddin M, Shelar A, Sgarali, Patel V, Khan T. Formulation and in vitro evaluation of floating microspheres of ketoprofen prepared by emulsion solvent diffusion method. Int J Applied Pharm2010; 2(1):76-82.
16.Belgamwar V, Shah VS, Surana J. Formulation and evaluation of oral mucoadhesive multiparticulate system containing metoprolol tartarate: Anin vitro – ex vivo characterization.Current Drug Delivery 2009; 6: 113-21.
17. Patrick BO, James WM. Preparation of microspheres by the solvent evaporation technique.Advanced Drug Delivery Reviews 1997;28: 25 –42.
18. Philip PM, Arya GS.Formulation andin vitroevaluation of nelfinavir mesylate microcapsules using cellulose acetate. Int J Pharma Pharm sci 2010;2(3): 169-71.
19. Patel J, Patel D, Raval J.Formulation and evaluation of propranolol hydrochloride-loaded carbopol-934P/ethyl cellulose mucoadhesive microspheres. Ira J Pharma Res 2010; 9 (3): 221- 32.
9 / Signature of the candidate:
10 / Remarks of the Guide: / The topic selected for dissertation is satisfactory. Adequate equipments and chemicals are available to carry out project work.
11 / Name and Designation of:

11.1 Institutional Guide: / Dr. Divakar Goli,
Prof&Principal,
Acharya & B.M. Reddy College of Pharmacy, Soladevanahalli,, Sarvepalli Radhakrishnan Road,Hesaraghatta Main Road, Bangalore-90.
11.2 Signature:
11.3 Co-Guide:
11.4 Signature:
11.5 Head of the Department: /

Dr. Shivanand Kalyanappa.

Professor & HOD
Dept. of Pharmaceutics
Acharya & B.M. Reddy College of Pharmacy,
Bangalore-560 090
11.6 Signature
12 / 12.1 Remarks of the Principal

Dr. Divakar Goli, Principal,

12.2 Signature Acharya& B.M Reddy college of Pharmacy

Bangalore-90

1