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8/13/2019 Bme07 Putnam
1/27
The Putnam Lab
Delivery of Nucleic
Acid-Based
Therapeutics
Biomaterial Design
and
Synthesis
Project Dist r ibut ions
Outer Membrane
Vesicle
Engineering
siRNA
plasmid DNA
Controlled Release
Bioadhesives
Biolubricants
Vaccines
Adjuvants
8/13/2019 Bme07 Putnam
2/27
Approachpolymeric libraries with serial changes in composition
side chain length/compositionbackbone composition(hydrophilicity/hydrophobicity)
molecular weight
side chain density
side chain terminicomposition/ratios
side chain length/compositionbackbone composition(hydrophilicity/hydrophobicity)
molecular weight
side chain density
side chain terminicomposition/ratios
Multifactorial biomaterial
design and synthesis
Formulation number
20
40
60
80
100
5 00 1 00 0 1 50 0 2 00 0 2 50 0 3 00 0 3 5 0 0 4 00 0 4 50 0
Refolding
yield
Formulation number
20
40
60
80
100
5 00 1 00 0 1 50 0 2 00 0 2 50 0 3 00 0 3 5 0 0 4 00 0 4 50 0
Refolding
yield
Quantify efficacy of
each unique structure
Endosome(pH ~ 5)
Nucleus
EndosomalEscape
Nuclear transport
1. Transcription
2. Translation
Protein
Biophysical and subcellular
characteristics
Goal: Quantitative, mechanistic understanding of transfection
8/13/2019 Bme07 Putnam
3/27
The Putnam Lab
Delivery of Nucleic
Acid-Based
Therapeutics
Biomaterial Design
and
Synthesis
Project Dist r ibut ions
Outer Membrane
Vesicle
Engineering
siRNA
plasmid DNA
Controlled Release
Bioadhesives
Biolubricants
Vaccines
Adjuvants
8/13/2019 Bme07 Putnam
4/27
Motivationthe human body is a monomer factory
New polymeric biomaterialsfrom metabolic synthons Investigate metabolic pathways
Identify interesting monomers
Akin to PLGA polyesters
http://www.science.gmu.edu/~gsudama/csi803s97/met2.gif
Rational/targeted selection
Engineered polymer properties
Synthetically challenging
8/13/2019 Bme07 Putnam
5/27
Dihydroxyacetone (DHA)
Scheme 1: Glycolysis pathway
Glucolytic metabolite
OH OH
O
Glucose
Pyruvic acid
DHA
8/13/2019 Bme07 Putnam
6/27
Dihydroxyacetone (DHA)
FDA approved for use in oral
and topical administration (the
active ingredient in sunless
tanning lotions).
Glucolytic metabolite
http://www.premiersalonsystems.com/
http://www.procyte.com/products/brand/asp/titanfoaming.shtml
Scheme 1: Glycolysis pathway
Glucose
Pyruvic acid
DHA
8/13/2019 Bme07 Putnam
7/27
O
O
O
II
O O
OH OH
OHOH I
III
O O
O O IV
O+
H3CO HV
n
O
O
O O
O
m
VI
O
O O
mVII
O
HO
HO
OH
OH
O
H3CO nO
H3CO nO
a
b
c
d
Zawaneh, P.; Doody, A.; Zelikin, A.; Putnam, D. Biomacromolecules (2006)
PEG-pDHASynthesis, characterization and application
O
O O
m
O
H3CO nO
Water soluble
block
Water insoluble
block
Postoperative adhesions
Seroma closure
Fistula blockade
Chemo-emboli
8/13/2019 Bme07 Putnam
8/27
DHA-based lipidsSynthesis
HYPOTHESISRelease slower with
increasing lipid
chain length
8/13/2019 Bme07 Putnam
9/27
DHA-based lipidsRelease kinetics
C8 C10
C12 C14
C16
8/13/2019 Bme07 Putnam
10/27
Career path and researchwhich have led you
Delivery of Nucleic
Acid-Based
Therapeutics
Biomaterial Design
and
Synthesis
Project Dist r ibut ions
Outer Membrane
Vesicle
Engineering
siRNA
plasmid DNA
Controlled Release
Bioadhesives
Biolubricants
Vaccines
Adjuvants
8/13/2019 Bme07 Putnam
11/27
Outer membrane vesicles (OMVs)natural vesicles for transfer of proteins and DNA
http://www.molbiol.umu.se/forskning/wai/
GOALEngineered vesicles to correctly fold
and stabilize proteins
Optimize antigen presentation to APCs
APPLICATIONSExpression/stabilization/delivery of
conformational antigens
Novel adjuvants to enhance existing
or poorly effective vaccinesOM
PG Per
Cyt
LPS
IM
Periplasmic
proteins are
entrapped within
the OMV lumen
Kuehn and Kesty (2005) Genes Dev19: 2645-55
8/13/2019 Bme07 Putnam
12/27
Section 2
Joint project with Neurological Surgery ChemoCoils and Brain Phantom
Creation and Validation of a Novel Drug Delivery Technique
Michael Shuler, Susan Pannullo, David Putnam, Jian Tan
8/13/2019 Bme07 Putnam
13/27
ChemoCoils and Brain PhantomCreation and Validation of a Novel Drug Delivery Technique
Cornell Cross-CampusNeurological Surgery/Biomedical Engineering Project
Michael Shuler, Susan Pannullo, David Putnam, Jian Tan
July 2007
8/13/2019 Bme07 Putnam
14/27
Reviewing the ProblemMalignant Gliomas
Highly aggressive braincancers
Recur locally need goodlocal control techniques
Only 1 validated/FDAapproved device:chemotherapy wafers Minimal survival benefit
Poor conformality to resection
cavity Minimal brain penetration
Submaximal dose
Only one drug (BCNU) delivered
Drug delivery poorly
understood
8/13/2019 Bme07 Putnam
15/27
Chemotherapy Coils and
Brain Phantom: The ProjectYear 1
Development of an in vitro BrainPhantombased upon MagneticResonance Imaging of humanbrain and brain tumor
Development of polymer coilswith appropriate mechanical,chemical, and drug releaseproperties.
Test, using dyes and IMAGING,the distribution, depth ofpenetration, and duration of
chemical dyes from differentpolymer formulations
Year 2 Refinement of delivery
system/drug mixtures
Animal trials
Clinical trials
HypothesisMaintaining contact with cavity wall will
improve treatment outcomes
8/13/2019 Bme07 Putnam
16/27
Controlled Release Polymer
Incorporate both p(CPP:SA) (poly
(carboxyphenoxypropane-sebacic acid) and
polyester of -caprolactone
Diameter and porosity are controlled byelectrospinning
Wafer: 14mm in diameter and 1mm thick
Mesh: interwoven fibers (
8/13/2019 Bme07 Putnam
17/27
Pressure Model
Brain experiences around 10mmHg of
pressure in brain cavity.
For our first experiments we will use a
simple water tank to create the pressure.
14cm
8/13/2019 Bme07 Putnam
18/27
Alternate Pressure Model
Agarose Brain
Silicone
Encapsulation
Pressure
Probe
Syringe to alterpressure
8/13/2019 Bme07 Putnam
19/27
Mathematical Model
Simulate the transport of drugs from variouspolymer constructs to the brain
Assumption: transport of drug occurs bydiffusion and convection (due to edema) withelimination (e.g. internationalization)
Goal: to predict drug concentration anddeduce drug penetration in the artificialtissue, then compare with our brain phantommodel
8/13/2019 Bme07 Putnam
20/27
Section 3
Bill Olbricht Microcatheters for drug delivery to the brain
8/13/2019 Bme07 Putnam
21/27
Microcatheters for Convection-Enhanced Delivery
Diffusion only gets you so far.
Convection can get you further.
8/13/2019 Bme07 Putnam
22/27
Remodeling ECM to enhance nanoparticle delivery
Channel 1: Deliver enzymes that degrade ECM components (hyaluronan and
chondroitan sulfate proteoglycans) to increase permeability OR hyperosmolar
solutions that swell interstitium to increase permeability
Channel 2: Deliver drug-laden nanoparticles to reduce drug elimination during
transport in tissue and extend release time
5 mm
8/13/2019 Bme07 Putnam
23/27
Flexible microfluidic catheters
8/13/2019 Bme07 Putnam
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Section 4
C. C. Chu
Materials for drug delivery
8/13/2019 Bme07 Putnam
25/27
C. C. Chu
Biodegradable hydrogels as cytokine (IL-12)carriers for immunotherapy of cancer.
Estone carrier from polysaccharides. Biodegradable carriers of nitric oxide
derivatives for nitric oxide biofunctionality.
Biodegradable hydrogels and microspheres asanticancer drug (e.g., Doxorubicin, Paclitaxel)carriers.
Biodegradable hydrogels as gene carriers
8/13/2019 Bme07 Putnam
26/27
Burst release followed by sustained releaseover 3 mo. wo/ bioactivity loss.
4 factors control IL-12 release charge,
hydrophilicity, gel crosslinking density,
biomaterial biodegradation.
water
Dry
1. Cytokine carriers for immunotherapy:
Interleukin 12 impregnated within Arginine-based biodegradable hydrogels.
2. Biodegradable carriers for nitric oxide derivatives (NOD):
a. NOD conjugated with biodegradable biomaterials.
b. NOD Impregnated within biodegradable microspheres or nanofibers.
2.5
0.5
2
1
1.5
0 0dN
itroxylradicalrelease(%
)
9d
10
2
6
2d 4d
conjugated
Phenylalanine-based
poly(ester amide)
microspheres w/
10 mg NO/g PEA
-
PGA, PLA, PEA
8/13/2019 Bme07 Putnam
27/27
3. Estrone carrier from polysaccharides:
a. Starch-estrone conjugate.
b. Dextran-estrone conjugate.
Polysaccharide OCCH2- Estrone
O pH 7.4
pH 8
4.Biodegradable hydrogels and microspheres as anticancer drug carriers:a.from poly(ester amide) gel and microspheres
b. from intelligent polysaccharide-synthetic hydrogels
P l ( t id ) l I lli D h i h d l
0
10
20
30
40
50
60
70
80
0 10 20 30 40 50 60 70 80 90 100
Time (days)
Cumula
tiverelease(%)
NDF-1
NDF-3
NDF-5
Doxorubicin release