bme07 putnam

8/13/2019 Bme07 Putnam

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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


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


3/27

The Putnam Lab

Delivery of Nucleic

Acid-Based

Therapeutics

Biomaterial Design

and

Synthesis


Outer Membrane

Vesicle

Engineering

siRNA

plasmid DNA

Controlled Release

Bioadhesives

Biolubricants

Vaccines

Adjuvants


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


5/27

Dihydroxyacetone (DHA)

Scheme 1: Glycolysis pathway

Glucolytic metabolite

OH OH

O

Glucose

Pyruvic acid

DHA


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


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


8/27

DHA-based lipidsSynthesis

HYPOTHESISRelease slower with

increasing lipid

chain length


9/27

DHA-based lipidsRelease kinetics

C8 C10

C12 C14

C16


10/27

Career path and researchwhich have led you

Delivery of Nucleic

Acid-Based

Therapeutics

Biomaterial Design

and

Synthesis


Outer Membrane

Vesicle

Engineering

siRNA

plasmid DNA

Controlled Release

Bioadhesives

Biolubricants

Vaccines

Adjuvants


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


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


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


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


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


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 (


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


18/27

Alternate Pressure Model

Agarose Brain

Silicone

Encapsulation

Pressure

Probe

Syringe to alterpressure


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


20/27

Section 3

Bill Olbricht Microcatheters for drug delivery to the brain


21/27

Microcatheters for Convection-Enhanced Delivery

Diffusion only gets you so far.

Convection can get you further.


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


23/27

Flexible microfluidic catheters


24/27

Section 4

C. C. Chu

Materials for drug delivery


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


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


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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

bme07 putnam

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