This presentation includes both protien and gene drug delivery. i think it will b helpful for both pg and ug students.

1. PRESENTED BY
SUBODH S SATHEESH
MPHARM
PHARMACEUTICS
ECPS
1protien drug delivery

2.  Rise as a potential drug
 Treatment of many diseases
 Recombinant technology
 protein market
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3.  Primary
 Secondary
 Tertiary
 quaternary
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4.  Classification by functions of proteins:
 Enzymes : DNA and RNA polymerase
 Hormones : Endorphine and encephalin
 Transport proteins : Cytochrome C, Albumin, Haemoglobin.
 Antibodies : Interferon, Fibrin.
 Structural proteins : Collagen, Elastin.
 Motor proteins : Actin, Myosin.
 Signalling proteins : GTPase.
 Storage proteins Egg albumin, milk casein.
 Classification of proteins by location in the living cell:
Membrane proteins Internal proteins External proteins Virus proteins
 Classification of proteins by post translational modification:
Native protein Glyco protein
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5.  Solubility
 Partition coefficient
 Self aggregation
 Hydrogen bonding
 Association
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6.  1.Denaturation
 2. Adsorption
 3. Aggregation and Precipitation .
 Chemical Instability:
 1. Deamidation .
 2.Oxidation and Reduction .
 3. Proteolysis
 4.Disulfide exchange
 5. Racemisation
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7.  Enzymatic barrier Limits absorption of protein drugs fr
om G.I. tract.
 Intestinal epithelial barrier Involved in the transport of
protein drugs across the intestinal epithelium
 Capillary endothelial barrier Involved in transport of pr
otein drugs across the capillary endothelium.
 Blood brain barrier
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8.  Parenteral systemic delivery:
 Non‐parenteral systemic delivery:
 a. Oral route
 b. Nasal route
 c. Buccal route
 d. Ocular routee.
 e. Rectal route
 f. Transdermal route
 g. Pulmonary route
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9.  Most effective method of delivery
 intravenous(IV), intramuscular(IM), subcutaneous(SC
 Biomedical applications
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10.  Insulin
 Lente semilente ultra lente
 Needles and infusion pumps
 Vasopressin
 Covering a section of microporous
polypropylene(Accurel) tubing with collodion
 a long lasting and constant in‐vitro release
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11.  a. Oral route
 b. Nasal route
 c. Buccal route
 d. Ocular routee.
 e. Rectal route
 f. Transdermal route
 g. Pulmonary route
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12.  Patient complaince
 Strategies
 Promote adsorption
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13.  Bonding of (PEG) and alkyl groups fatty acid
radicals to produce desired amphiphilic oligomers
 oligomers are conjugated to proteins or peptides to obtain desi
red amphiphilic products
 can resist excessive degradation of protein or peptide drugs
 technology reduces self‐association, increases penetration and
increases compatibility with formulation ingredients than pa
rent drug
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14.  protection against the metabolic barrier in GIT
 absence of a carrier system for absorption of peptides
with more than three amino acids
 Proteins are labile due to susceptibility of the pepti
de backbone to proteolytic cleavage
 Prodrug approach
 olefenic substitution, d‐amino acid substitution,
dehydro amino acid substitution, carboxyl reduction
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15.  Pulmonary protein delivery offers both local targeting for the
treatment of respiratory diseases and increasingly appears
to be a viable option for the delivery of proteins systemi
cally
 The lung is easy to access, has decreased proteolytic acti
vity compared with the gut
 Careful choice of carrier and device can facilitate deliver
y to a specific area of the lungs.
 The only protein for inhalation currently available on the
market is Dnase
 Inhaled insulin leuprolide and gamma‐interferon are in
trials.
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16.  Insulin: The nasal absorption of insulin is increased by co
administration of bile salts
 inhibition of respiratory virus infection was studied
by intranasal administration of human leukocyte
interferon.
 Nasaldeliveryof oligopeptides:
 Examples:
Dipeptides: 1‐tyrosyl‐1‐tyrosine and its methyl esters
 Tripeptides: Thyrotropin‐releasing hormone(TRH)
 Pentapeptides: Leucin‐enkephalin, met‐enkephamide
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17.  Advantages of nasal route:
 Convenient,simple,practicalway of drug administration
 The high vascularization permits better absorption.
 First pass metabolism can be avoided.
 Rapid onset of action.
 Disadvantages of nasal route:
Long term use may lead to toxicity to mucosa.
During disease states (e.g. common cold) some altera
tion in the nasal environment may occur
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18.  it offers excellent accessibility and avoids degradation
of proteins and peptides.
 Various types of polymers like sodium CMC, hydroxypropyl
methyl cellulose, PVP, acacia,calcium carbophil are used for
delivery of proteins or peptides via buccal route.
 1)Adhesive tablets: e.g. Adhesive tablet based on hydroxy
propylcellulose.
 2)Adhesive gels: e.g. By using polyacrylic acid and poly
methacrylate as gel forming polymers.
 3)Adhesive patches: e.g. Protirelin in HEC patches and buserel
in
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19.  It is robust, much less sensitive to irreversible irrit
ation even on long term treatment.
 Absence of enzymatic barrier.
 Well acceptable to the patients.
 Easy accessibility administration as dosage forms. It is
attached or removed without any pain or discomfort
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20.  The ocular route holds immense potential for peptides or prote
ins intended for pathological ophthalmologic conditions
 The ocularroute is the site of choice for the localized delivery
of opthalmologically active peptides and proteins for the
treatment of ocular disease that affect the anterior segment
tissues of eye.
 The use of nanoparticles, liposomes, gels, ocular inserts, bioad
hesives or surfactants are necessary to enhance ocular absorpt
ion of proteins or peptides
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21.  Affect aqueous humor dynamics:
Calcitonin gene related factors, LHRH, vasopressin
 Immunomodulating activities: Cyclosporine, interfer
ons.
 Act on inflammation : Substance P, enkephalins.
 Affect wound healing:Epidermal growth factor, fibro
nectin
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22.  It is one of the recent ideas
 The coadministration of an absorption promoting adjuvant
s such as sodium glycocholate, has been reported to enhan
ce the rectal absorption of insulin.
 Bile salts, such as sodium salts of cholic, deoxycholic and glyc
ocholic acids, have also been shown to enhance the rectal ab
sorption of insulin
 Vasopressin and its analogs,pentagastrin and gastrin, ca
lcitonin analogs
and human albumin have been investigated for rectal de
livery of protein or peptide based pharmaceuticals
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23.  It is highly vascularized.
 It avoids first pass or presystemic metabolism.
 Drug can be targeted to the lymphic system.
 It suitable for drugs that cause
nausea/vomiting and irritate GI mucosa on oral admini
stration.
 A large dose of drugs can be administered.
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24.  Transdermal delivery has attracted considerable interest
as a route for administering peptides and proteins.
 The small peptides such as thyrotropin releasing hormone (
TRH) vasopressin, have great difficulty in permeating the ski
n barrier.
 Percutaneous absorption of elastin peptides has shown better
distribution.
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25.  Iontophoresis
It is use of electric current to drive charged drug molecules
into skin by placing them under an electrodeof like charged
 DC iontophoretic device, as the power source for direct current an
d were able to deliver insulin transdermally to diabetic hairless
rats, with attainment of a reduction in hyperglycemia
 Pulse DC iontophoresis: By delivering a pulse current with a 20
% duty cycle (4µsec), followed by an 80% depolarizing period(16µs
ec), a β‐blockers was successfully delivered systemically human
subjects without polarization induce skin irritation.
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26.  In this method ultrasound is applied via a cupling conta
ct agent to the skin.
 Insulin, IFN γ, erythropoietin can be delivered by this method
 Surfactants and azone have been used for topical delivery of p
eptide or proteins
 Prodrugs; Prodrug with modeled physicochemical characte
ristic permeated well across the skin. LHRH, TRH, neurotens
in can be delivered by this method.
 Penetrationenhancers: Penetration enhancers like oleic acid,
dimethylsulphoxide are used.
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27.  Advantages of transdermal route:
 Avoids the hepatic first‐pass effect and gastrointestinal breakdown.
 Provides controlled and sustained administration particularly
suitable for the treatment of chronic disease.
 Reduces side‐effects, often related to the peak concentrations of the circulating
agent;
 Enables self‐administration and improves patient compliance, due to its conve
nience and ease of use.
 Permits abrupt termination of drug effect by simply removing the delivery sys
tem from the skinsurface.
 Limitations of transdermal route:
 A low rate of permeation for most of protein drugs due to their large molecular
weight.
 High intra‐ and inter‐patient variability. Because the skin has a relatively low pr
oteolytic activity, the peptide drugs have poor skin permeability.
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28. GENE DRUG DELIVERY
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29.  successful design of a gene delivery system requires
complete understanding of the interaction mechanism
between the target cell and delivery system
 Cell targeting refers to delivery of the therapeutic agent
to a specific compartment or organelle of the cell
 Endocytosis gene therapy
 cellular release takes place to initiate DNA
transcription and translation, and to produce the related
protein
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30. 30protien drug delivery

31.  Viral drug delivery
 Non viral drug delivery
 Physical methods
 Chemical methods
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32.  It consist of viruses that are modified to be replication-
deficient which were made unable to replicate by redesigning
which can deliver the genes to the cells to provide expression
 Viral systems have advantages such as constant expression and
expression of therapeutic genes
 Limitations are use of viruses in production, immunogenicity,
toxicity and lack of optimization in large-scale production.
 current gene technologies concentrates on the use of viral
vectors that provide high transduction effectiveness and
advanced level of gene expression.
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33.  Adenoviral systems
 Retroviral systems
 Lentiviral systems
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34.  Adenoviruses (Ad) were first discovered in 1953 by
isolation from human adenoid tissue cultures.
 Commonly used as gene vectors
 adenoviruses Ad2 and Ad5 are The most widely studied
adenoviruses.
 The capsid of an adenovirus determines virus tropism.
 Adenoviruses are well-characterized, non-integrated, 26–40
kb in length, relatively large, non-enveloped, linear dsDNA
viruses coated with icosahedral particle, with a diameter of
950 Å (excluding elongated fiber proteins) and a molecular
weight of approximately 150MDa
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35.  in Figure it was reported that crystal structures of single
Ad proteins contained fiber knob, shaft, domains, penton
base, hexon, and cysteine protease.
 Ad capsid consists of 252 sub-units called capsomeres,
which contain 240 hexonproteins and 12 of the penton
base.
 In addition, the capsid contains pIIIa, pVI, PVIII, and
pIX proteins. Each of the 12 capsid corners contains
penton bases wrapped by 5 hexons.
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36.  Instead of combining its own DNA with the genomes of the
host cell, the adenovirus, remains as an episome within the
infected cell.
 Penton and fiber proteins of virus capsid interact with the
coxsackievirus-adenovirus receptor cell surface protein to
provide cell binding
 Viral capsid proteins dissociate prior to endocytosis, and the
pH value of the viral endosome decreases due to proton pumps
 For successful delivery of DNA to the nucleus, viruses must
facilitate cell-specific binding, endocytosis internalization,
propagation from endocytic vesicles to cytosol, delivery into
cytoplasm
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37.  Retroviruses are diploid, single-stranded, circular-enveloped
RNA viruses of the family Retroviridae, with a genome of 7–
11 kb, and a diameter of approximately 80–120 nm
 Retroviruses cause diseases such as AIDS, leukemia, and
cancer
 Retroviruses are viruses that integrate with host genome to
produce viral proteins (gag, pol, env) that are extracted during
gene delivery.
 Commonly used retroviruses are the Moloney murine
leukemia virus species, which have the capacity to deliver
exogenous genetic material up to approximately 9 kb.
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38.  An ideal retroviral vector for gene delivery should be cell-
specific, regulated, and safe.
 Retroviruses have a lipid envelope. In order to enter a host
cell, they use the interactions between cellular receptors and
virally encoded proteins, which are embedded in the
membrane
 CKRs are a family of cell-sfurface-G-proteins functioning as
receptors to stain molecules called chemokines
 Retroviruses introduce their genetic material to the host cell
genome in a stabile manner during mitotic division
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39.  Most retroviruses infect cells that can be actively divided
during mitotic division. This property protects normal tissue,
and although it naturally targets the tumor
 A retrovirus infects the target cell by providing interaction
between viral envelope protein and cell surface receptor on the
target cell
 Many types of retrovirus types require degradation of mitosis
and then the nuclear envelope for the arrival of a viral genome
within the nucleus.
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40.  Lentivirals are viral systems without small, retrovirus-like
viral proteins and no capacity for replication
 The most important advantage of lentiviruses compared with
other retroviruses is their ability for gene transfer to non-
dividing cells
 Genome of lentiviruses have a more complicated structure;
they contain accessory genes which regulate viral gene
expression, control combination of infectious particles,
modulate viral replication in infected cells, and are associated
with the continuance of infection.
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41.  HIV-1 is one of the most widely used lentiviral vectors, and
contains six accessory genes (tat, rev, vif, vpr, nef, vpu). These
proteins are involved in all steps of cell cycles, which are
termed: budding, maturation, and integration.
 Lentiviral vectors do not require degradation of the nuclear
membrane for integration.
 Lentiviruses that are encoded with the Gag matrix protein
integrase enzyme and vpr protein interact with the nuclear
import mechanism of the target cell and manage active
transport of pre-integration complex via nucleopores.
 Receptors have been defined for many retroviruses. The best-
characterized example is CD4 molecule, which serves as a
receptor for lentiviruses including HIV.
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42.  many studies that used viral vectors reported unsatisfactory
results, due to the immunologic and oncogenic adverse effects
of these vectors.
 It is overcome by NVGDS
 non-viral vectors have many advantages, such as easy of
fabrication, cell/tissue targeting, and low immune response
 biggest disadvantage of non-viral vectors in clinical use is low
transduction efficiency.
 the biggest difficulty in gene therapy is the development of
physical methods to ensure gene transfer to target cells of the
gene delivery vectors and delivered gene.
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43. 43protien drug delivery

44.  Compared to viral delivery systems, non-viral carriers are less
toxic and 450 immunogenic non-viral vectors is ease-of-
production.
 A number of barriers need to be overcome in order to increase
the effectiveness of non-viral vectors in humans. These
barriers are classified as production/formulation/storage;
extracellular barriers; and intracellular barriers
 Anatomic barriers are extracellular matrixes coating the cells,
which prevent direct transport of macromolecules to target
cells through epithelium and endothelial cell sequences
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45.  The most critical barrier to effective DNA transfection was
regarded as the transition of plasma membrane. Typically,
naked nucleic acids cannot cross cell membrane by cellular
uptake mechanisms such as endocytosis, pinocytosis, and
phagocytosis
 Physical approaches, including electroporation, gene gun,
ultrasound, and hydrodynamic delivery are based on the
application of a force to increase the permeability of the cell
membrane and promote intracellular gene transfer
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46.  Two of the most important advantages of synthetic
carriers are that they do not display immunogenicity, and
large-scale production is easy
 Non-viral vectors can trigger an inflammatory response,
since they do not provide a specific recognition
 they are much less dangerous than viral vectors in terms
of antigen specific immune response
 Non-viral vectors should be designed according to
specific cell targeting; cellular uptake and and potential
immune response should be minimized.
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47.  Gene guns
 Electroporation
 Ultrasound
 polymers
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48.  Delivery with gene gun method is also termed ballistic DNA
delivery or DNA-coated particle bombardment, and was first
used for gene transfer to plants in 1987.
 This method is based on the principle of delivery of DNA-
coated heavy metal particles by crossing them from target
tissue at a certain speed
 Generally, gold, tungsten or silver microparticles were used as
the gene carrier
 Gene-gun-based gene transfer is a widely tested method for
intramuscular, intradermal and intratumoral genetic
immunization.
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49.  It does not use toxic chemicals or complex biological
systems
 delivery is achieved without the need for a receptor
 DNA fragments of various sizes, including large ones,
are transported, there is no need to introduce foreign
DNA or protein.
 it has high repeatability, production of heavy metal
particles is easy .
 However, in this method, gene expression is short-term
and low.
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50.  Ultrasound has many clinical advantages as a gene delivery
system, due its easy and reliable procedure
 Microbubbles or ultrasound contrast agents decrease cavitation
threshold with ultrasound energy.
 Mostly perfluoropropane-loaded albumin microbubbles were
used.
 The transfection efficiency of this system is based on
frequency, time of ultrasound treatment, the plasmid DNA
mount used, etc
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51.  Electroporation includes controlled electric application to
increase cell permeability
 Electroporation introduces foreign genes into the cell by
electric pulses. In this method, pores are formed on the
membrane surface to enable the DNA to enter the cell.
 If the molecule is smaller than the pore size , it can be
transferred to the cell cytosol through diffusion
 loaded molecules and ions can be transported from the
membrane via electrophoretic and electro-osmotic means via
the effect of electric regions
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52.  Polymers are long-chained structures composed of small spliced
molecules called monomers.
 Polymers that are composed of a repeated monomer are called
homopolymers, while those composed of two monomers are called
copolymers.
 Biodegradable polymers are non-water soluble, and undergo
chemical or physical change in biologic environment.
 Polyamides, dextran, and chitosan are examples of biodegradable
polymers
 non-biodegradable polymers are not degraded in biological
environments;
 hydrophilic polymers are hydrogels, which are non-water soluble
and swell in water, while hydrophobic polymers are non-water
soluble and do not swell.
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53.  hydrophilic hydrogel polymers include PVA, polyvinylacetate,
polyethyleneglycol, polyacrylic acid.
 hydrophobic polymers include silicones, and polyethylene
vinyl acetate
 ethyl cellulose (EC), hydroxypropyl methyl cellulose
(HPMC), cellulose acetate phthalate (CAP), and eudragit
derivatives are commonly used in controlled release systems
 For polymer selection, in addition to its physicochemical
characteristics, characterization of extensive biochemical
characteristics and preclinical tests are required to demonstrate
its reliability.
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54.  Encapsulation
 Electrostatic interaction
 Liposomes
 dendrimers
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55.  Most of the polymeric vectors presently in use form
complexes with negatively charged DNA by
electrostatic interaction
 At adequate nitrogen– phosphate ratio, the polymer and
the DNA form nanocomplexes, which allows both
cellular DNA uptake and also protects the DNA from
nuclease enzyme.
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56.  An alternative to electrostatic condensation of DNA is
encapsulation of DNA with a biodegradable polymer.
 Polymers that have an ester linkage in their structures (like
polyesters) are hydrolytically degraded to short oligomeric and
monomeric compounds, which are more easily discharged
from the body.
 The degradation mechanism and DNA release can be
controlled by changing the physicochemical characteristics
and composition of the polymer.
 DNA is protected from enzymatic degradation by
encapsulation.
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57.  Remington the science and practice of pharmacy
volume 1
 Nejm Gene therapy and novel drug delivery page 1-36
 Intechopem Gene therapy and viral and nonviral
vectors 387-402
 Ijcpr protien drug delivery volume 3 285-327
 Rhienberg Protien drug delivery 1-17
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58. 58protien drug delivery