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CHAPTER -1
INTRODUCTION:
Because the brain is tightly segregated from the circulating blood by a unique
membranous barrier, the blood-brain barrier (BBB), many pharmaceuticals cannot be
efficiently delivered to, or sustained within the brain; hence, they are ineffective in
treating cerebral diseases. Therefore, drug delivery methods that can provide brain
delivery, or eventually preferential brain delivery (i.e. brain targeting), are of
particular interest.
To achieve successful delivery, an understanding of the major structural, enzymatic, and
active transport aspects related to the BBB, and of the issues related to lipophilicity and its
role in CNS entry, is critical. During the last years, considerable effort was focused in the
field of brain-targeted drug delivery. Various more or less sophisticated approaches, such as
intracerebral delivery, intracerebroventricular delivery, intranasal delivery, BBB disruption,
nanoparticles, receptor mediated transport (vector-mediated transport or ‘chimeric’
peptides), cell-penetrating peptides, prodrugs, and chemical delivery systems, have been
attempted. These approaches may offer many intriguing possibilities for brain delivery and
targeting, but only some have reached the phase where they can provide safe and effective
human applications. Site-target indexing and the use of targeting enhancement factors can
be used to quantitatively assess the site-targeting effectiveness from a pharmacokinetic
perspective of chemical delivery systems.
Drug targeting is the delivery of drugs to receptors or organs or any other specific part of the
body to which one wishes to deliver the drug exclusively. The drug therapeutic index(TI)
as measured by its pharmacological response and safety. Drugs minimizing its interaction
with non target tissue. The desired differential distribution of drug by its targeted delivery
would spare the rest of the body and thus significantly reduce the over all toxicity while ma
intaining it's therapeutic benefits. The targeted or site specific delivery of drugs is indeed a
very attractive goal because this provides one of the most potential ways to improve the the
rapeutic index of the drug. The need for targeted delivery of drugs is best illustrated with
peptide drugs where failure in the clinic may not be due to a poor intrinsic activity, but
rather due to transport factors including widespread disposition,rapid catabolism and excre
tion, variable or in efficient extravasations, and the subsequent high dosing levels required
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to obtain a therapeutic effect (Tomlinson etcal.1986). Earlier work done between late 1960s
and the mid 1980s'stressed the need for drug;-carrier systems primarily to alter the
pharmacokinetics of the already proven drugs whose efficacy might be improved by alter-
ing the rates of metabolism in liver or clearance by the kidneys (Pozanski&Juliano, 1984).
These approaches generally were not focused to achieve site-specific or targeted delivery
such as getting a cytotoxic drug to cancerous tissue while sparing other normal, though
equally sensitive tissue (Papahadjopoulos, 1978)1.
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3. Active targeting (Ligand mediated targeting and Physical targeting)
4. Dual targeting
5. Double targeting
6. Combination targeting
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(1)First order targeting (organ compartmentalization).
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(5) PHYSICAL TARGETING (TRIGGERED RELEASE):
The selective drug delivery programmed and monitored at the external level (ex vivo) with
the help of physical means is referred to as physical targeting; In this mode of targeting,
some characteristics of the bioenvironmental are used either to direct the carrier to a
particular location or to cause selective release of its contents . The first such approach
reported is the temperature sensitive liposomes, which were developed and applied to
tumour by (Weinstein and co-workers, 1979).
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Combination targeting for the site- specific delivery of proteins and peptides these
targeting systems are equipped with carriers, polymers and homing devices of molecular
specificity that could provide a direct approach to target site. Modification of proteins and
peptides with natural polymers, such as polysaccharides, or synthetic polymers, such as
poly (ethylene glycol), may alter their physical characteristics and favour targeting the
specific compartments, organs or their tissues within the vasculature 3.
1.2.1 ENDOCYTOSIS:
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functions. It is the main cellular activity involved in the internalization of the extracellular
cargo and their vesicular coat proteins, which are subsequently processed via different
pathways to appropriate intracellular targets. Phagocytosis is the engulfment of the
endogenous and exogenous particulate materials, such as bacteria, erythrocytes, latex beads,
colloidal particles and immunoglobulin molecules. It is performed by the phagocytic cells of
the hepatic sinusoids, the tissue fixed macrophages (histocytes) and the blood macrophages
or monocytes that the fluid phase and receptor mediated pinocytosis are not separate cellular
events, but they are different facets of the same event. Non specific adsorption piocytosis is
responsible for the uptake of many non-glycosylated proteins particularly following cellular
damage or protein denaturation.
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1.3.2 LUNG AS A TARGET ORGAN:
The lung possesses the next highest levels of nearly all the metabolic enzymes found in
liver an in some cases even higher specific activities in certain cell types. Additionally, in
contrast to all other tissues, the lung receives total venous return first, so it in an ideal
position to regulate the concentration of substrates in the blood before they reach the
arterial circulation hence avoiding problems that may be associated with a hepatic first pass
and permitting a more efficacious sequestration of a drug entity.
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circulation making it difficult to attain adequate drug concentration at the target organ/cell
after the compound has absorbed via the portal blood Reduction in local gastrointestinal
irritation and toxicity .An overall modulation in the rate of drug input thus providing a
sustained delivery5.
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CHAPTER-2
REVIEW OF LITERATURE:
2.1 LIPOSOMES:
Liposomes have attracted a considerable amount of intevest for potential use as a drug
delivery system owing to their suitable characteristics. They consist of one or more
concentric phospholipid bilayers enclosing an aqueous space. They are biocompatible,
biodegradable, and normally nonimmunogenic. More importantly, they are capable of
loading both hydrophilic and hydrophobic drugs in the aquesous and bilayer phase
respectively. Drugs encapsulated in liposomes are protected from enzymatic degradation
and other inactivation processes. There are basically two different modes in liposome
targeting passive and active targeting. The former takes advantage of the fact that
systemically injected liposomes are rapidly and efficiently taken up by phagocytic
cells of the reticuloendothlial system (RES) located Mainly the live and the spleen Some
of the advantage of liposome are as follows:
• Provides selective passive targeting to tumour tissue.
• Increased efficacy and therapeutic index.
• Increased stability via encapsulation
• Reduction in toxicity of the encapsulated agent.
• Site avoidance effect.
• Improved pharmacokinetic effects (reduced elimination, increased circulation life
times)
• Flexibility to couple with site-specific lignads to achieve active targeting
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medium the molecules in self assembled structures are oriented in such a way that the
polar portion of the molecule remains in contact with the polar environment and at the
same time shields the non-polar part. Among the amphiphiles used in the drug delivery
viz. soaps, detergents, polar lipids, the latter (polar lipids) are often employed to form
concentric bilayered structures. However, in aqueous mixture these molecules are able
to form various phases, some of them are stable an others remain in the metastabel
state. At high concentrations of these polar lipids, liquid-crystalline phases are formed
that upon dilution with an excess of water can be dispersed into relatively stable
colloidal particles. The macroscopic structures most often formed include lamellar,
hexagonal or cubic phases dispersed as colloidal Nanoconstructs (artificial
membranes) referred to a liposomes, hexasomes or cubosomes , respectively.The most
common natural polar phospholipids are phosphatidylcholine (PC). These are amphipathic
molecules in which a glycerol bridge links to a pair of hydrophobic acyl hydrocarbon
chains with a hydrophilic polar head group, phosphocholine explains that the fatty chains
are embedded in the hydrophobic inner region of the membrane, oriented at an angle to
the plane of the membrane surface, the hydrophilic head group, including the phosphate
portion. Points out towards the hydrophilic aqueous environment. Molecules of PC are not
soluble (rather dispersible) in aqueous media in the physical chemistry sense, as in
aqueous media they align themselves closely in planer bilaryer sheets to minimize the
unfavorable interactions between the bulk aqueous phase and long hydrocarbon fatty acyl
chain. Such interactions are completely eliminated when the sheets fold over themselves
to form closed, sealed and concentric vesicles. The large free energy change between an
aqueous and hydrophoble environment explains the most favored orientation of lipids to
assemble as concentric bilayer structures that exclude confrontation between aqueous and
hydrophobic domains.Thus the amphipathic (amphiphilic) nature of Phospholipids and
their analogues render them the ability to form closed concentric bilayers in the presence of
water. Liposomes (lipid vesicles) are formed when thin lipid films or lipid cakes (of
amphiphilic nature) are hydrated and stacks of liquid crystalline bilayers become fluid and
swell.
2.1.2 METHODS OF LIPOSOME PREPARATION:
Liposomes are manufactured in majority using various procedures in which the water
soluble (hydrophilic) materials are entrapped by using aqueous solution of these materials
as hydrating fluid or by the addition of drug/drug solution at some stage during the
manufacturing of liposomes (Ostro, 1987, 1989; Talsma and Cromunelin, 1992). The
lipid soluble (lipophilic) materials are solubilized in the organic solution of the constitutive
lipid (s) and then evaporated to a dry drug containing lipid film followed by its hydration.
These methods involve the loading of the entrapped agents before or during the
manufacturing procedure (passive loading ) However, certain types of compounds with
ionizable groups,
then membrane phospholipids are disrupted, they can reassemble themselves into tiny
spheres, smaller than a normal cell, either as bilayers or monolayers. These are liposomes.
The lipids in the plasma membrane are chiefly phospholipids like phosphatidyl
ethanolamine and cholesterol. Phospholipids are amphiphilic with the hydrocarbon tail of
the molecule being hydrophobic; its polar head hydrophilic. As the plasma membrane faces
watery solutions on both sides, its phospholipids accommodate this by forming a
phospholipid bilayer with the hydrophobic tails facing each other.
Liposomes can be composed of naturally-derived phospholipids with mixed lipid chains
(like egg phosphatidylethanolamine), or of pure surfactant components like DOPE
(dioleoylphosphatidylethanolamine). Liposomes, usually but not by definition, contain a
core of aqueous solution; lipid spheres that contain no aqueous material are called micelles,
however, reverse micelles can be made to encompass an aqueous environment.
Various chemical analysis methods used for quantitative and qualitative tests of liposomal
components prior to and after the preparation are critical characteristics of liposomes
(Barenholz and Cromellin 1994). These methods become more essential to characterize
liposomes, which require lipid stability cropping up from oxidatin, lipid peroxidation,
hydrolysis and degradation ni various environment used in their manufacturing6.
1. Chemical Characterization
Phospholipid concentration Lipid phosphorus content using Barlett assay
Stewart assay, HPLC
Cholesterol concentration Cholesterol oxidase assay and HPLC
Phospholipid peroxidation UV absorbance. TBA ( for endoperoxidase),
iodometric (for hydroperoxidase) and GLC.
Phospholipid hydrolysis HPLC and TLC and fatty acid concentration
Cholesterol auto-oxidation HPLC and TLC
Anti- oxidatant degradation HPLC and TLC
pH pH meterr
Osmolarity Osmometer
2. Physical Characterization
Vesicle shape and surface morphology Transmission electron microscopy , freeze-fricture
electron microscopy
Vesicle size and size distribution Dynamic light scattering, transmission electron
Submicron range microscopy, zetasizer Transmission electron
Micron range microscopy,
2.2 NANOPARTICLES:
The colloidal earners based on biodegradable and biocompatible polymeric systems have
largely influenced the controlled and targeted drug delivery concepts. It was realized that
the nanoparticles loaded bioactives could not only deliver drug (s) to specific organs within
the body but delivery rate in addition could be controlled as being by standers, burst,
controlled, , pulsatile or modulated. The possibilities and potentials further prompted the
work and as a result a great deal of related information covering preparation
methodologies, characterization. Engineering, bio-fate and toxicology has been gathered.
The understanding that relates to the biodistribution in particular has propelled and
motivated the development of functionally designed nonoparticulates6.
The selection of the appropriate method for the preparation'of nanoparticles depends on
the physicoehemical characteristics of the polymer. Various Proteins and Polysaccharides
used for the prepartion of Nano particles:
Proteins Polysaccharides
Gelatin Alginate
Albumin Dextran
Lectins Chitosan
Legunin Agarose
Vicilin Pullulan
The drug to loaded. on the contrary, the preparation techniques largely determine the inner
structure, in vitro release profile and the biological fate of these polymeric delivery systems
.Two types of systems with different inner structure are apparently possible matrix type
system consisting of entanglement of oligomeror polymer units
(nanoparticles/nanoshpheres). A reservoir type of system comprised of an oily core
surrounded by an embryonic polymeric shell (Nanocapsules). The Polymers are strictly
structured to a nanometric size range particle (s) using appropriate methodologies.
FIG-2.3:NANOPARTICLE
2.2.2 CHARACTERIZATION OF NANOPARTICLES:
The nanoparticles are generally characterized for size, density, electrophoresis mobility,
angle of contact and specific surface area.
The particle size is one of the most important parameters of nanoparticles. Particles size
and sizing of sub-optical particulates is a different procedure, as it involves not only
procedural variability, but some of the surface associated properties may even change
during sizing procedure. Two main techniques are being used to determine the particle size
distribution of nanopartitcles and include photon correlation spectroscopy (PCS) and
electron microscopy. The latter (TEM) and freeze -fracture techniques. The size evaluation
of nanoparticale dispersion demonstrates better results with freeze-fracturing microscopy
and photon correlation spectroscopy as quantitative methods. The freeze-fracturing with
poly (methyl methacrylate) is confronted with and interrupted by in process
particles aggregation which only yields a few discrete particles for size measurement of
analysis. The electron microscopy however, could be adopted as an alternative option,
which measures individual particles for size and its distribution. It is relatively less time
consuming. Additionally, freeze fracturing of particles allows for morphological
determination of freeze fracture procedures, TEM permits differentiation among
nanocapsules, nanoparticles and emulsion droplets. Similarly, scanning electron
microscopy is much less time consuming. However, since particles are based on organic
and non-conductive material, they require from 30-50 nm. Thus determined size should be
denoted as gold- coated particle size rather than as particle size.
Table2.2: Different Parameters and Characterization Methods for
Nanoparticles:
Parameter. Characterization Method (S)
Particle size and size distribution Photon correlation Spectroscopy (PCS)
laser defractometnnry transmission electron microscopy
scannming Electron Microscopy.
The use of small and perfectly round Microspheres with exactly the same size circumvents
all of the disadvantages that are encountered while using powders and granulates. These
Microspheres are free-flowing and roll with practically no friction, that means there is no
abrasion, guaranteeing a dust-free environment.
In this work, computational fluid dynamics (CFD) techniques are utilized to study
invasive drug delivery in multi-dimensional brain geometries with the consideration
of the chemical interactions that the drug undergoes while it diffuses into the brain
tissue. A challenge is to accurately reconstruct the three-dimensional structure of the
human brain. We are able to resolve very accurately the brain geometry and render
physiologically consistent the distribution of the complex brain inner organization.
We distinguish between gray and white matter and assign transport properties of
relevance according to the data obtained by MR images or histological data. We will
quantify with numerical simulations the diffusive and convective transport
phenomena in the porous brain tissues and the effectiveness of the drug release to a
desired region. This approach will help to evaluate precisely the penetration depth of
the drug and the concentration profiles need to surpass set thresholds in order to
ensure proper efficacy of the drug. We rigorously examine the variables that influence
CED and pose constraints to the treatment. These include effect of infusate (bulk)
flow rate, concentration of the infusate, drug diffusivity, effect of molecular weight of
the drug, and effect of white matter anisotropy, infusate leak-back by considering
metabolic uptake by the parenchyma cells and re-absorption of the bulk fluid [2].
Understanding the parameters that could possibly influence the convective delivery of
drugs in the CNS is very important because, it will improve the current medical
approaches. The proposed methodology will provide a systematic approach to
optimally choose catheter dimensions, infusion rates, drug concentrations etc. The
information obtained from these accurate simulations could be used to model inverse
kinetic problems capable of predicting the mass diffusivity of the drug and the kind of
metabolism that actually takes place.
The term transcranial route means the brain targeted transfer of drug molecules across
the cranium through the layers of the skin and skin appendages of the head, arteries
and veins of the skin of the head, the cranial bones along with the diploe, the cranial
bone sutures, the meninges and specifically through the emissary veins. The
administration of drugs through the scalp in ayurvedic system for the diseases
associated with the brain was evaluated with a view to develop a novel targeted route
for central nervous system drugs. It is expected to circumvent the systemic side
effects of oral route. Diazepam was dissolved in an oil medium and applied on scalp
as practiced in the ayurvedic system. Thirty rats were tested on the rotating rotarod for
muscle relaxant effect of diazepam. Five groups of rats tested were the control,
diazepam i.v. injected (280 µg/0.1 ml) group, two groups treated with transcranial
diazepam oil solution (1.5 mg/0.2 ml) and the transcranial blank vehicle treated
groups. Holding time in triplicate for each rat on the rotating rotarod was measured.
The holding times following each treatment was statistically compared (one-way
ANOVA). The pooled average times for the control, diazepam i.v. injected, diazepam
oil solution transcranial treated two groups and the blank vehicle treated groups were
35.45, 4.73, 16.5, 15.39 and 33.23 seconds respectively. The two groups subjected to
the brain targeted transcranial route showed a statistically significant decrease (50%
drop) in the holding time against the control group indicating the centrally acting
muscle relaxant effect due to absorption of diazepam into the brain through the
proposed route.
Man entertained a special care in all matters relating to the head because the head
housed the brain. The effects of a bath are remarkably different from that of a body
wash. A head injury, however trivial, is considered a precarious situation and a pimple
on the face may be fatal. The apprehensions mentioned above and the drug delivery
route that is being undertaken in the present study is incidental to a special anatomical
feature of the skull. The emissary veins draining blood from extracranial sites into the
intracranial sinuses pierce a series of foramina present in the cranial bones. Seven
major sinuses within the skull are interconnected by a number of anastomosing veins,
which finally drain intracranially into the jugular veins giving ample scope for the
diffusion of the drug molecules into the nerve tissue of the brain. There are thirteen
emissary veins connecting extracranial sites of the head with the intracranial
sinuses[1]. The emissary veins are present in all higher animals starting with aves[2]
and their presence in the horse is well established.
The arteries of the scalp send small twigs to the underlying bones of the scalp. The
spongy diploe within the flat skull bones is also well supplied by numerous small
diploic branches from arteries both on external and internal surfaces of the skull[3].
These anatomical arrangements of the vascular system are made use of in the
investigations to develop the brain targeted transcranial route (abbreviated TCR) of
drug delivery.
In the ayurvedic system there are five methods, namely Shirodara, Shiroabyanga,
Shiropichu, Shirovasthi and Shiropralepa in which drugs are delivered by the
transcranial route[4]. Most of these ayurvedic preparations are oil based. There are
many household ayurvedic medicinal head oils for minor ailments such as headaches,
sinusitis, vertigo and migraine. An important dosage design feature in this study is the
use of the essentially non polar active diazepam drug moiety dissolved in an oil
medium as against the use of polar salt forms in aqueous media that are popular with
the modern formulations. Diazepam was selected as the screening agent since it is a
prototypical benzodiazepine acting in the central nervous system. Diazepam has
central depressant and centrally acting skeletal muscle relaxant effects[5]. The skin
area of the head which drains venous blood through emissary veins into the
intracranial locations is probably the region lying above the circular contour drawn
through the angles of the mouth and the ears. Therefore the venous blood draining the
eyes, ears and the nose are also drained by this route.
The proposed transcranial route is intended to circumvent the side effects encountered
during treatment by the oral route. Some of the central nervous system diseases such
as epilepsy need long term therapy. There are prospects of adopting the transcranial
route for several groups of drugs. They include antiepileptics, antipsychotics,
tranquilizers, analeptics, antiparkinsonian drugs, those acting on the endocrine glands
located in the brain, drugs related to diseases of the labyrinth, glaucoma,
anticoagulants and those employed in the treatment of drug addiction. CNS side
effects of other drugs could be counteracted by administering the specific antagonists
through transcranial route. There is a good prospect in adopting the transcranial route
in veterinary practice as well. Development of subscalpal injections is another
possibility. The proposed brain targeted transcranial route of drug delivery could be
viewed as a parallel drug delivery system to that of metered dose inhalers in diseases
of the respiratory system.
Ethical clearance for animal experiments was granted by the Ethical Review
Committee of the Medical Faculty, Colombo. Both male and female inbred Sprague-
Dawley rats weighing between 165-230 g were used. They were divided into five
groups of six rats including three from either sex. In the animals that were subjected
to the treatment by transcranial delivery route the hair of the scalp was trimmed with
scissors without injuring the skin. It was done within the trapezoidal area of the head
bound by the pair of eyes and ears, closer to the line joining the ears than the eyes.
They were kept at room temperature (around 30°) and fed with adult rat meal pellets
and water without any restriction. They were appropriately numbered with picric acid.
The equipment was devised by having an electrically driven horizontal circular rod 18
mm in diameter, 29 cm in length rotating constantly at 15 rpm, mounted on two side
panels 43 cm above the base. The rod was covered with no. 300 silicon carbide
abrasive paper to provide roughness for the rats to grip the rod firmly.
Blank oil vehicle 0.2 ml for TCR administration was prepared by dissolving 1 ml of
isopropyl alcohol in 3 ml of sesame oil. Disposable insulin syringes of 100 units/1 ml
capacity with 29 gauge needle were used for rat dorsal tail vein i.v. injection as the
positive control and also to deliver the oil based formulation drop wise onto the scalp
of the animals.
The animals were divided into the following five groups. Group 1, untreated animals
(blank). Group 2, reformulated diazepam tail vein i.v. injected animals (positive
control). Group 3, transcranial diazepam oil solution treated animals tested 15 minutes
after drug application (test group A). Group 4, same as Group 3 but tested 45 minutes
after drug application (test group B). Group 5, transcranial blank vehicle treated
animals tested 15 minutes after the application (control group).
One animal at a time was placed on the rotating rod. Rats fell off the rod when the
grip was lost. The holding time on the rod in seconds was observed for each animal.
Each rat was subjected to three rotarod trials, five minutes apart.
Group 2 was tested on the rotarod 15 minutes after the i.v. injection. Group 3 (test
group A) and Group 4 (test group B) were tested as follows. Each rat was treated with
0.2 ml of the transcranial diazepam oil formulation containing 1.5 mg of diazepam
using insulin syringe. The oil solution in 1/3 quantities were delivered drop wise on to
the hair trimmed area of the scalp leaving a gap between the needle end and the skin
in three stages as follows. First application at 00:00 time, 2nd application at 00:15
minutes and 3rd application at 00:45 minutes. The first application was followed by
gentle rubbing on the scalp with a gloved finger previously smeared in the diazepam
oil formulation by stroking ten times in a cranial to caudal direction. This was to
facilitate dispelling any air pockets and to bring the oil solution into intimate contact
with the skin of the scalp.
Each animal was tested thrice 5 minutes apart on the rotating rotarod starting 15
minutes after the 3rd application in Group 3 and starting 45 minutes after the 3rd
application in Group 4, respectively. Accordingly animals were tested one hour after
the first application in Group 3 and one and a half hours after the first application in
Group 4. In testing Group 5 the animals were treated with blank oil solution similar to
Group 3 and tested 15 minutes after the 3rd application of the vehicle. Statistical data
analysis was done using SPSS software package. One-way ANOVA and Dennett T 3
Post Hoc test was done to compare mean holding times between the groups.
Holding time on the rotating rotarod:
Mean value of three trials were calculated for each animal in all five groups. The
individual means of time in seconds on the rotating rotarod of six animals in each
group were pooled to get the mean holding time on the rod for each group [Table - 1]
The mean holding times on the rotarod for group 1 (untreated) and the transcranial
blank vehicle treated group (Group 5) being not significantly different (35.45 and
33.23 s, respectively) suggests that there are no effects of either isopropyl alcohol or
sesame oil or scalp stroking on the holding time and therefore on the muscle
tone/grip. The diazepam i.v. injected group 2 as expected had a significant effect on
the mean holding time, bringing it down to 4.73 seconds, further proving the known
muscle relaxant effect of diazepam.
The two groups subjected to transcranial route treatment with diazepam showed
statistically significant reduction in mean holding times from that of the untreated and
the vehicle treated groups and this reduction was almost by 50% [Figure - 1]. These
results suggest that the diazepam drug molecules have been conveyed transcranially
in to the nerve tissue of the rat brain under the experimental conditions described
here. However the mean holding times of the transcranially treated groups were
longer than in the i.v. treated group and these differences were statistically significant,
suggesting that the amount of diazepam delivered by the transcranial route to the CNS
is significantly lower than that delivered by the i.v. route. This difference in diazepam
delivery may be due to the novel experimental route and the properties of the oily
formulation of diazepam prepared for transcranial application.
The results of the experiments further indicate that after a certain point irrespective of
the concentration of the drug in the oil solution, the volume applied and the time
allowed before testing the response to the drug tend to even out unlike in the
conventional oral or parenteral routes. This is evident by the fact that the holding
times are nearly the same for two transcranially treated groups despite the
substantially longer time allowed for the group 4 to effect the diffusion of the drug.
There appear to be a wide therapeutic window for diazepam administered by the
transcranial route.
Scientists began to study targeted drug delivery, because the traditional drug delivery
system had many disadvantages, such as high toxic effect and high minimum effective
dose. In traditional drug delivery system, after the patient takes some drugs, the drugs
will distributed throughout his body through the systemic blood circulation. Only a
small amount of drugs can reach the affected organ which it needs to act on. Since
many drugs have some toxicity, they can kill some helpful bacteria or normal cells in
some normal organs.
The targeted drug delivery system can overcome these shortcomings and deliver the
drugs right to the specific organ, without having any adverse effects on other healthy
organs and tissues. Actually, the targeted drug delivery can be used to cure many
diseases, such as the cardiovascular diseases and diabetes. However, the most
important application of the targeted drug delivery is to treat the cancerous tumor.
There are two kinds of targeted drug delivery. The first one is active targeted drug
delivery, such as some antibody drugs. The second one is passive targeted drug
delivery, such as the Enhanced Permeability and Retention effect (EFR-effect). Some
Important applications are given below.
Magnetic drug targeting allows the concentration of drugs at a defined target site
generally and importantly, away from the reticular endothelial system (RES) with
the aid of a magnetic field. Site-directed drug targeting is one way of local or
regional antitumor treatment. The drug & an appropriate Ferro fluid are formulated
into a pharmaceutically stable formulation which is usually injected through the
artery that supplies the target organ or tumor in the presence of an external
magnetic field. Prolonged retentions of the magnetic drug carrier at the target site
alleviate or delay the RES clearance & facilitates extra vascular uptake. For
effective retaining of magnetic drug carrier, the magnetic forces must be high
enough to counteract liner flow rates within the organ or tumor tissue (between 10
& 0.05 cm/s depending on vessel size & branching pattern . There is increase in
drug concentration in the target tissue after administration of the drug dose has
been observed.The efficiency of chemotherapy treatment may be enhanced to a
great extent by magnetically assisted delivery of cytotoxic agent to the specific
site. There are a large number of magnetic carrier systems which demonstrates
increasing drug concentration efficiency at the tumor site.
Magnetism can play very important role in cancer treatment. The first clinical
cancer therapy trials using magnetic microspheres were performed by Lubbe et al.
in Germany for the treatment of advanced solid tumor while current preclinical
research is investigating use of magnetic particles loaded with different
chemotherapeutic drugs such as mitoxantrone, paclitaxel. Non invasive permanent
magnetic field for one hour way found to induces lethal effects on several rodent &
human cancers. Anticancer drugs reversibly bound to magnetic fluids & could be
concentrated in locally advanced tumors by magnetic field that or arranged at
tumor surface outside of the subject.
A magnetic fluid has been reported to which the drugs, cytokines & other molecule
can be chemically bound to enable that agent to be directed within subject under
the influence of high energy magnet. In one of such examples magnetic
doxorubicin in liposome, significant anticancer effect in nude mice bearing colon
cancer .
(2) Magnetic bioseparation:
Heat treatment of organs or tissues, such that the temperature is increased to 42–46
C and the viability of cancerous cells reduces, is known as hyperthermia. It is
based on the fact that tumor cells are more sensitive to temperature than normal
cells. In hyperthermia it is essential to establish a heat delivery system, such that
the tumor cells are heated up or inactivated while the surrounding tissues (normal)
are unaffected.
a)Intracellular hyperthermia:The alternative approach is to use fine particles as
heat mediators instead of needles or rods such that hyperthermia becomes
noninvasive. When fluids containing submicron-sized magnetic particles(typically
1–100nm) are injected, These particles are easily incorporated into the cells, since
their diameters are in the nanometer range. These magnetic particles selectively
heat up tissues by coupling AC magnetic field to targeted magnetic nano particles.
As a result, the whole tumor can be heated up uniformly This is called intracellular
hyperthermia. It has been shown that malignant cells take up nine times more
magnetic nano particles than normal cells. Therefore the heat generated in
malignant cells is more than in normal cells. Also, as blood supply in the cancerous
tissues is not normal, the heat dissipation is much slower. Hence, the temperature
rise in the region of tumor is higher than in the surrounding normal tissues. It is
therefore expected that this therapy is much more concentrated and localized.
c) Combination therapy: There also exists the combination therapy which would
induce hyperthermia treatment followed by chemotherapy or gene therapy. A
combination of chemotherapy or radiation therapy with hyperthermia is found
much more effective than hyperthermia itself. The approach involves use of
magnetic carriers containing a drug to cause hyperthermia using the standard
procedure, followed by the release of encapsulated drug that will act on the injured
cells. It is anticipated that the combined treatment might be very efficient in
treating solid tumor. Several reasons are given for the enhanced effect. Tumors are
poorly vascularised and it can be hard for therapeutic agents to reach their target.
Heat increases the perfusion of a tumor and therefore drugs are transported more
effectively into the target tissues. In addition, heat makes blood vessels more
permeable to drugs. This occurs preferentially in tumors where blood vessels tend
to be structurally incomplete. On the other hand, normal blood vessels are
surrounded by a basement membrane and other perivascular cells and not
significantly affected by heat. It has recently been reported that hyperthermia
increases the rate of liposome leakage into tumors by a factor of 2–5 depending on
the type of tumor. In normal tissues however, enhancement of liposome leakage is
not reported.
Malignant brain tumours can arise in one of two ways. On the one hand, astrocytes
undergo genetic changes accompanied by upregulation of certain receptors, such as
the platelet-derived growth factor (PDGF), endothelial growth factor receptor (EGFR)
or vascular endothelial growth factor (VEGF). These progressive changes culminate
in the formation of a glioblastoma. On the other hand, most primary glioblastomas
arise de novo, without the need for gradual progression from an astrocytoma to a
high-grade astrocytoma to a glioblastoma multiforme.
Brain tumors are one of the most lethal forms of cancer. They are extremely difficult
to treat. Although, the rate of brain tumor incidence is relatively low, the field clearly
lacks therapeutic strategies capable of overcoming barriers for effective delivery of
drugs to brain tumors. Clinical failure of many potentially effective therapeutics for
the treatment of brain tumors is usually not due to a lack of drug potency, but rather
can be attributed to shortcomings in the methods by which a drug is delivered to the
brain and into brain tumors. In response to the lack of efficacy of conventional drug
delivery methods, extensive efforts have been made to develop novel strategies to
overcome the obstacles for brain tumor drug delivery. The challenge is to design
therapeutic strategies that deliver drugs to brain tumors in a safe and effective manner.
This review provides some insight into several potential techniques that have been
developed to improve drug delivery to brain tumors, and it should be helpful to
clinicians and research scientists as well.
CHAPTER-5
CONCLUSION:
The blood brain barrier (BBB) and the systemic toxicity of conventional
chemotherapy present obstacles to the success of future blood-borne drug therapies of
brain tumors. The work with polymer-encapsulated cancer drugs suggests an
alternative and more focused treatment approach. Our experimental strategy integrates
direct intracerebral drug delivery, sustained drug release from liposomes or polymer
implants, and increased targeting of the drug either by chemically modifying the drug
or by using tumor-specific carriers. This review will present some of the recent work
on targeted drug delivery for brain cancer treatment.
Cancer is one of the most challenging diseases today, and brain cancer is one of the
most difficult malignancies to detect and treat mainly because of the difficulty in
getting imaging and therapeutic agents across the blood-brain barrier and into the
brain. Many investigators have found that nanoparticles hold promise for ferrying
such agents into the brain [20-22]. Apolipoprotein E was suggested to mediate drug
transport across the blood-brain barrier [23]. Loperamide, which does not cross the
blood-brain barrier but exerts antinociceptive effects after direct injection into the
brain, was loaded into human serum albumin nanoparticles and linked to
apolipoprotein E. Mice treated intravenously with this complex induced
antinociceptive effects in the tail-flick test. The efficacy of this drug delivery system
of course depends upon the recognition of lipoprotein receptors. Kopelman and
colleagues designed Probes Encapsulated by Biologically Localized Embedding
(PEBBLE) to carry a variety of unique agents on their surface and to perform multiple
functions [22]. One target molecule immobilized on the surface could guide the
PEBBLE to a tumor. Another agent could be used to help visualize the target using
magnetic resonance imaging, while a third agent attached to the PEBBLE could
deliver a destructive dose of drug or toxin to nearby cancer cells. All three functions
can be combined in a single tiny polymer sphere to make a potent weapon against
cancer. Another anti-cancer drug, doxorubicin, bound to polysorbate-coated
nanoparticles is able to cross the intact blood-brain barrier and be released at
therapeutic concentrations in the brain [24]. Smart superparamagnetic iron oxide
particle conjugates can be used to target and locate brain tumors earlier and more
accurately than reported methods [25]. It is known that folic acid combined with
polyethylene glycol can further enhance the targeting and intracellular uptake of the
nanoparticles. Therefore, nanomaterial holds tremendous potential as a carrier for
drugs to target cancer cells.
It is very difficult for the medicine to destruct the target organism at the point of infection
because of complex cellular network of an organism. Target delivery of drugs, as the
name suggests, is to assist the drug molecule to reach preferably to the desired site. The
inherent advantage of his technique has been the reduction in the dose and side effects of
the drug.
By virtue of their size smaller than that of blood capillaries, intravenously administered
particulate drug carriers get accumulated in the liver cells. Among the particulate drug
carriers liposome's are a potential mode of delivery for the treatment of intracellular
infections as the cells of mononuclear phagocytic system easily take these up.
Microparticles may serve as future mode of delivery for the drugs of protein nature .
Orally delivered micro particles (<5 urn in size ) are taken up by the peye's patches. This
leads to induction of immune response against the antigen released from the
microparticles. Also, the antigen is protected from the loss of activity in the Gl tract. A
major limitation is the effective uptake of these particles from the Gl tract which is even
less than 1% However, a combination of biological approach such as incorporation of
specific ligands on the surface of surface of these particles enhances their uptake.
Magnetic Vesicular systems have been realized as extremely useful carrier systems in
various scientific domains. Over the years, magnetic microcarriers have been
investigated for targeted drug delivery especially magnetic targeted chemotherapy due
to their better tumor targeting, therapeutic efficacy, lower toxicity and flexibility to be
tailored for varied desirable purposes
CHAPTER-6
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