Effect of glibenclamide drug supported to chitosan and chitosangelatin matrices in blood glucose of Rattus rattus (Sprague dawley)
Rafid M. A. Hassan Wasfi : college of pharmacy kufa university, Hanaa Jaafer
jabaar AL –kabee College of Pharmacy, kufa university , Sami AL-maliki:college of
education basrah university,Salah shaker AL-Luaibie:college of scince basrah
university
Abstract
The purpose of this study is to developing controlled drug delivery
system for glibeclamide release from Chitosan and chitosan gelatin
matrices . The hypoglycemic drug glibenclamide supported to chitosan
and chitosan –gelatin matrices as drug delivery system were prepared
.The swelling rate of both matrices in simulated gastric fluid (SGF) and
intestinal fluid (SIF) were measured. The blood glucose of glibenclamide
supported to chitosan and chitosan –gelatin matrices in laboratory rats
were measured after(0,2,4,6,8,24 hr) of treatment.
The result showed that the swelling rate of both matrices was increased
specially in SGF compared with SIF and the swelling rate in chitosan gelatin matrix is higher than chitosan matrix .The results proved the
ability of glibenclamide supported to chitosan and chitosan–gelatin
matrices to significantly reduction p≤0.01 of blood glucose during the
period of treatments compare with control group.
INTRODUCTION
The ways in which chemicals or drugs are administered have gained
increasing attention in the past two decades. Normally, a chemical is
administered in a high dose at a given time only to have to repeat that
dose several hours or days later. This is not economical and sometimes
results in damaging side effects. As a consequence, increasing attention
has been focused on methods of giving drugs continually for prolonged
time periods and in a controlled fashion .The primary method of
accomplishing this controlled release has been through incorporating the
chemicals within polymers. In the pharmaceutical field, in addition to
the importance of polymers, The skin, the gastrointestinal tract, the nose
and the eye are of particular importance.The polymeric constituted
serves simply as drug carrier and is usually chosen because of its
biological inactivity. [1].biodegradable polymers have been used simply
as inert carrier vehicles. However, the effective delivery of new drug
therapies, including peptides, proteins, and genetic- and cell-based
drugs, places greater demands on the performance of the polymer plate
form. [2].Polymeric materials are essential during the system
development as matrix or carrier which can slowly release or deliver the
drug to an appropriate position. Synthetic polymers carriers chosen as
potential drug carriers must exhibit certain properties which are listed
chales et al. [3].whose suggest that the polymer should be non toxic,
must be insoluble and easy to be synthesized, it must exhibit a narrow,
definite molecular weight distribution,It should provide attachment
release sites or the possibility of the incorporation of drug–polymer
linkage, both sites and linkages must display controlled stability,The
polymer should display the ability to be direct to predetermined cell
types, either by its inherent physical-chemical properties or the
incorporation of specific residues.It should be compatible with the
biological environment.,Ideally, it should be biodegradable or
eliminated from the organism after having fulfilled its function. There
are many miscellaneous materials that can be used in controlled drug
delivery preparation such as polycarbophil, malete anhydride, , xanthan
gum, pectin, cyclodextrin and the fruit, seeds, pods of natural gums,
mucilage and chitosan polymer [4].This study suggest chitosan and
gelatin polymer and glibenclamide which is hypoglycemic drug as a
model for controlled drug delivery system. Chitosan is widely used in
medical and pharmaceutical applications such as biomaterial,
anticoagulant, antibacterial, anticholestrolemic[5], antiulcer,antiuricemic
and wounds healing[6].It is used in pharmaceutical industries as carrier
in control drug delivery systems [7; 8; 9; 10;11]. The other important
polymer in drug delivery system is Gelatin which is a heterogeneous
mixture of proteins derived from animal collagen by thermal
denaturation of collagen [12].It is not found naturally it is typically
obtained by boiling bovine, fishes, pigs and ox skin and bones with very
diluted acid it can also be extracted from fish skin in fact the ward
gelatin is derived from Latin gelatos meaning frozen or stiff [13]. It also
has a range of industrial and medical application , gelatin is an
ingredient in film coatings , medical devices such as artificial heart
valves and in specialized meshes used to repair wounds. Although
gelatin has been touted for decades as a good source of protein and great
nail and hair strengthen, for the most part there is a little hard to proof to
support the vague health claims made for gelatin products [14].
Methods
1-preparation of glibenclamide-chitosan matrices
Chitosan is natural polysaccharide was prepared by the partial
deacylation of chitin obtained from shrimps[15].Solid tablets of
glibenclamide [Daonil] Medochem Ltd, Limassol-cyprus [Europe]
Company brought from the local market. The drug was isolated accorddin
to[16]. then chitosan solution was prepared by dissolving (2% w/v) of
chitosan powder in 100 ml of 0.1N acetic acid with stirring. Then 100 mg
of glibenclamide were added with stirring for 1hr at room temperature.
Gluteraldelyde were then added to the mixture in the ratio 1ml/100ml as
cross linkage agent [17]. 100ml of 0.1 M of sodium hydroxide was added
to the mixture . The mixture was filtered and washed in distilled water
until pH changed to 7 then dried using air dried technique [18].
2-Preparation of glibenclamide- chitosan- Gelatin matrices
Chitosan 1.6 gm were dissolved in 100 ml of 0.1 N acetic acid.Then
gelatin polymer 0.4 gm (Applihem company) were added to the mixture
with stirring. 100 mg of glibenclamide were added with stirring until the
mixture mixed very well for 1hr at room temperature. Gluteraldelyde
1/100ml was added. Then 100 ml of 0.1N sodium hydroxide was added to
the mixture . The mixture was filtered and washed in distilled water until
pH changed to 7 then dried using air dried technique. Simulated gastric
and intestinal fluid was prepared according to [19]
3- Determination of the Swelling Rate of Chitosan and ChitosanGelatin Matrices in SGF and SIF:The Swelling rate of chitosan and chitosan- gelatin matrices were
measured by Swollen 200 mg and 10 mm diameter disk in 50ml of
simulated gastric fluid and. Simulated Intestinal fluid with out adding
digestive enzyme. The weight of the disk was determined at time interval
for 4 hrs. The Swelling rate were Calculated according to [20]. As the
following
Q= WS- WD / WD
Q=swollen rate
Ws=swollen weight
WD= dry weight
4-Experimental Investigation of Oral Administration of
Glibenclamide Supported to Chitosan and Chitosan- gelatin
matrices .
Forty eight Adult male rats weighting 200-225gm 12- 16 weeks old are
used in this experiment. The animals were randomly divided to four
groups each of 8 rats as the following:-
1-The First group was treated with 0.5gm/kg of glibenclamide supported
to chitosan matrix
2- The second group was treated with 0.5 gm / kg of chitosan Matrix as
control group.
3-The third group treated with 0.5 gm /kg of glibenclamide supported to
chitosan-gelatin matrix.
4- The fourth treated with chitosan –gelatin matrix as control group
5- The fifth group was treated with 0.5 ml of distilled water served as
control group.
6-The last group was treated with 2.5 mg glibenclamide drug as blind
Control group.
5- Collection of Blood Sample:Blood samples were collected at zero [as fasting and 2, 4,6,24 hrs after
oral administration] and Prior to killing. The animal must be food
deprived and drinking water was then replaced by glucose solution 20%
to preveut hypoglycemia [21].Blood glucose levels were assayed by using
enzymatic colorimetric test [GOD-PAP] using a standard kit [22]
Result and discussion
1- Determination of swelling rate of chitosan and chitosangelatin matrices in SGF and SIF buffer:
It is evident from figures( 1,2) show that chitosan and chitosan gelatin
matrices have the ability to swollen this property may be due to the
ability of chitosan to from the hydrogel compound which is three
dimensional polymeric material characterized by high ability to absorbed
a large a mount of water in the solutions.[23].The hydrogel compound
usually have a hydrophilic homogenous group, usually non soluble in
water because of the presence of chemical linkage by covalent or ionic or
physical by [crystallites] or hydrogen bounds between the chain of the
matrix [24]. As it appears from the present result that the swelling rate of
both matrices in SGF is higher than the swelling rate in SIF media The
differences in the swelling rate indicate thot bath matrices have Ph
sensitivity these result agreed with [18]. who founds that chitosan have a
higher swelling rate in the acidic media. The swelling activity may also
be due to the porosity of the network of both matrices. chitosn and
chitosn- geltin matrices did swell extensively in SGF media due to the
cationic properties of chitosn polymer. It appears from the present results
that chitosan- gelatin matrix has swollen rate more than chitosan matrix
and this may be due to the presence of gelatin in the matix which has the
hydrogel ability of gelatin to for NH3 ions in the acidic media which lead
to increase bounding with water by hydrogen bound which in turn
increased the water uptake of the matrix [25].
2-Evaluation the hypoglycemic action of glibenclamide drug
and the drug supported to chitosan and chitosan-gelatin
matrices
It appears from the present results shown in figures (3, 4, 5) that there is a
significant decreasing effect on blood glucose level in the treated group
with glibenclamide drug and the drug supported to chitosan and chitosangelatin matrices.This decreasing effect refers to the hypoglycemic activity
of the drug because glibenclamide drug described as active hypoglycemic
drug in human [26]. the drug has Also the same ability to reduce blood
glucose level in Laboratory animal such as laboratory rats [27].Generally
the mode of action of glibenclamide drug is by the activation of receptors
on the beta cells of pancreatic islets to release stored insulin in response
to glucose level also the action of glibenclamide drug is well described
by[28] these authors have revealed that of the islet cell is freely
permeable to glucose via the Glut2 transporter, and the glucose is
phosphorylated by the high- Km glucokinase. Therefore the blood
glucose concentration determines the flux through glycolysis, , so the
drug cause the inhibition of ATP sensitive K+ channels. The drug may
also enhance insulin action on, liver, muscles and adipose tissue by
increasing insulin receptor number and by enhancing the post receptor
complex enzyme reaction mediated by insulin. The result of that is
decreased hepatic glucose output and increasing glucose uptake in muscle
[26]. It also appears from the result there are a significant decreasing
effect in blood glucose level 2hr after treatment with the drug and the
drug supported to chitosan and chitosan gelatin matrices compared with
control groups the decreasing in blood glucose explain the release of the
drug from the matrices. The releasing of the drug occur because of the
swelling activity of both matrices. It appears from the result the
hypoglycemic activity of chitosan is more than chitosan- gelatin matrix at
2hr this may due to the release ratio of the drug in chitosan matrix is
higher than the release ratio of chitosan- gelatin in at this period. While
with the continuous of the matrices swelling the releasing of the drug is
increased and the blood glucose level is decreased. [29] found that the
release ratio of hydroquinone was controlled by the polymer cross linking
density and the degree of swelling rate of hydro gel matrix. The present
results also showed a significant decreasing effect on blood glucose level
in the treated group with the drug supported to chitosan and chitosangelatin matrices in 24hr after treatment compared with control group and
the group treated with glibenclamide drug which showed an increasing in
blood glucose level at the same time i.e. 24 hr after treatment. This
increasing could be explained that the drug may eliminated from the
blood under normal elimination pathway of the drug and the body
regulate blood glucose level because the half life of glibenclamide is 10
hrs[26] while the matrices may provide a better control of the drug
release and prolong its action. This result agreed with [30;31] who found
that the drug supported to matrices may prolong the drugs action.
30
The swelling
rate
25
20
15
chitosan
chtosan gelatin
10
5
0
0
50
100
150
200
250
Time (min.)
Fig [1] The swelling rate of chitosan and chitosan -gelatin matrices in
SGF buffer
The swelling 10
rate
9
8
7
6
chitosan
5
chitosan gelatin
4
3
2
1
0
0
50
100
150
200
250
Time ( hr )
Fig(2) the swelling rate of chitosan and chitosan -gelatin matrices in
SIF buffer
Fig(3)Effect of glibenclamide supported to chitosan matrix in blood
glucose level of male rats
glucose level 200
(mg/100 ml)
150
control
100
chitosan gelatin
drug
50
0
chitosan gelatin drug
0
2
4
6
24
Time
Fig.(4) Effect of glibenclamide supported to chitosan- gelatin
matrix on blood glucose level [ mg /100 ml ] of male rats
glucose level
180
(mg/i00 ml) 160
140
120
100
80
60
40
20
0
Drug
chitosan drug
chitosan gelatin drug
0
2
4
6
24
Time (hr)
Fig (5) Comparison the effect of glibenclamide drug and the drug
supported to chitosan and chitosan- gelatin matrices on
blood glucose level of male rats.
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‫تاثيرعقار ‪ glibenclamide‬المسند على سبائك الكيتوسان والكيتوسان‪-‬جالتين في‬
‫مستوى السكرفي مصل دم الجرذان‬
‫)‪Rattus rattus (sprague dawley‬‬
‫الخالصة‪:‬‬
‫الغرض من الدراسة الحا لية هو لتطوير نظام للتحرر الدوائي المقنن لعقار ‪glibenclamide‬‬
‫الخافض للسكر من سبائك الكيتوسان والكيتوسان‪-‬جالتين ‪.‬تم قياس نسبة انتفاخ كال السبيكتين في‬
‫سائل المعدة االفتراضيي [‪ ]SGF‬وسيائل اممعياا االفتراضيي [‪. ]SIF‬كميا تيم قيياس مسيتوى‬
‫سييكر الييدم فييي الجييرذان المختبرييية المعامليية عقييار ‪ glibenclamide‬المحمييل علييى سييبائك‬
‫الكيتوسييان والكيتوسييان‪-‬جالتييين عييد [‪ ] 0,2,4,6,8,24‬سيياعة ميين المعامليية ‪.‬النييرج النتييائ ان‬
‫نسبة االنتفاخ قد ازدادج فيي كيال السيبيكتين وةاصية فيي سيائل المعيدة االفتراضيي مقارنية سيائل‬
‫اممعاا االفتراضي وان نسبة االنتفاخ في سبيكة الكيتوسان جالتين كانت اعلى من نسيبة االنتفياخ‬
‫في سبيكة الكيتوسان ‪.‬اثبتت النتائ قدرة عقار‪ glibenclamide‬المحمل على سبائك الكيتوسان‬
‫‪ p≤0.01‬لمسييتوى السييكر ف يي ال يدم ةييالف فتييراج‬
‫والكيتوسييان‪-‬جالتييين للخفييض المعنييو‬
‫المعاملة مقارنة مجموعة السيطرة‪.‬‬