Ligand mediated Targeting

Targeting components which have been studied and exploited are pilot molecules themselves
(bioconjugatres) or anxhored as kigands on some delivery vehicle (rug-carrier system). All the carrier
systems, explored so far, in general, are colloidal in nature. They can be specifically functionalized
using various biologically relevant molecular ligands including antibodies, polypeptides,
oligosaccharides (carbohydrates), viral proteins and fusogenic residues. The ligands afford specific
avidity to drug carrier. The engineered carrier constructs selectively deliver the drug to the cell or
group of cells generally referred to as target. The cascade of events involved in ligand negotiated
specific drug delivery is termed as ligand driven receptor mediated targeting.

Ligand mediated active targeting could be achieved using specific uptake mechanisms such
as receptor dependent uptake of natural low density lipoproteins (LDL) particles and synthetic lipid
microemulsions of partially reconstituted LDL particles coated with the apoproteins. The apoprotein
coat serves as a ligand for the LDL receptors expressed in the body. The ability of an immunoglobulin
coated carrier to promote its accelerated interception by the liver and spleen, possibly via pathways
involving Fc or C3B receptor mediated uptake, is one of the approaches that exemplifies and signifies
active targeting. Recently, Balley and co-workers, 1997 described active targeting of the protein-
coated liposomes using different targeting approaches: a two-step targeting approach and a direct
approach. The study was based upon the specificity of biotin binding to avidin r streptavidin.
Biotinylated molecules can be targeted in complex mixtures by using the appropriate avidin or
streptavidin conjugates with carrier system. In the two step targeting approach, the first step
prelabels target cells with biotinylated antibodies specific for a ligand on the target cell. At a
predetermined time that was sufficient was allow for plasma elimination of free antibody,
streptividin conjugated liposomes specific for biotin on the antibody prelabelled cells were
administered. The active targeting is referred to as two step targeting approach for the fact that it
requires the administration of the both the intermediate protein and liposomes separately, the
former o allow accumulation and binding to target cells, And the latter to bind with the intermediate
protein labeled target cells. The direct targeting approach exploits specificity by using liposomes
linked with targeting ligands via a biotin-avidin bridge.

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 tumor by Weinstein and co-workers, 1979. The
release of drug from temperature sensitive liposomes in the vicinity of a tumour (temperature status
higher or equal to the phase transition temperature of constitutive lipids) is brought by serum
components mostly the lipoproteins, which at phase transition induce released of the entrapped
drug.

It has been suggested that weakly anionic drugs, e.g. methotrexate, be released from
liposomes preferentially at low pH regions of tumours. Yatwin and co-workers, 1980 have reported
pH sensitive liposomes for selective release of content at low pH. This approach further modified
with antitumour antibodyanchoredas a site directing ligand with pH sensitive components of the
liposomes (pH sensitive immunoliposomes). The approach was found exceptional for tumour
targeting as well as cytosolic delivery of entrapped drugs or genetic materials (fig.2-4). In another
approach, the application of external magnetic field has been suggested for localization of magno-
responsive liposomes and microsphers within a preselected capillary bed.

Dual Targeting

This classical approach of the drug targeting employs carrier molecules, which have their wn
intrinsic antiviral effect thus synergies the antiviral effect of the loaded active drug. Based on this
approach, drug conjugates can be prepared with fortified activity pH sensitive Cytosolic Delivery
contents.

Double targeting

For a new future trend, drug targeting may be combined with another methodology, other than
passive and active targeting for drug delivery systems. The combination is made between spatial
control and temporal control of drug delivery. This combination is represented in fiugre2-5.

The temporal control of drug delivery has been developed in terms of control drug release prior to
development of drug targeting. If spatial targeting s combined with temporal control release results
in an improved therapeutic index by the following two effects. First, if drug release or activation is
occurred locally at therapeutic sites, selectively is increased by multiplication of the spatial selectivity
with the local release/activation. Second, the improvement in the therapeutic index by a
combination of a spatially selective delivery and a preferable release pattern for a drug, such aas
zero order release for a longer time period of the drugs. As shown in figure 2-5 when these two
methodologies are combined, it may be called “ Double Targeting”.

In order to achieve a double targeting effect site specifically of the drug, by virtue of
targeting moiety, a high specificity module (mainly a photosensitizer) is linked to antibodies. Mew
and co-workers in a series of studies (1983, 1985) on such double getting systems reported
haematoporphyrin (Hp)-anti-M-1 antidy jugated for the suppression tumour following cent light
exposure. Further, p-monoclonal antibody conjugates against a leukemia –associated antigen was
shown to have selective phototoxicity. Conjugates of monoclonal antibody and photosensitizers
were prepared using a spacer arm in order to circumvent t coupling drug d antibodycules (Oserofft
al., 1986).

Similar double targeting systems were reported different combinations of photosensitizer and
antibody specific either to a particular antigen or to cell lines expressing cell specific receptors.

Combination Targeting

Petit and Gombtz, 1998 have suggested the term combination targeting for the site-specific delivery
of proteins and peptides. These targeting systems are equippes 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 glycil), may alter their physical characteistics and favour targeting the specific
compartments , an or their tissues within the vasculature. Further vectorization of these modified
proteins and peptides into vesicular or microparticulate carriers may take advantage of the intrinsic
or inherited (through homing devices) properties of carrier to achieve site specific active targeting of
encapsulated contents. Some of the strategies are depicted in figure 2-6.

Problems associated with targeted delivery systems

Several problems have been identified which require alterations in targeting strategies particularly,
in vivo. These include:

 Rapid clearance of targeted systems specially antibody targeted carriers

 Immune reaction against intravenous administered carrier systems
 Target sue heterogeneity
 Problems of insufficient localization of targeted systems into tumour cells
 Down regulation and sloughing of surface epitopes
 Diffusion and redistribution of released drug leading to no-specific accumulation.