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Coming to Madison

Pre-”hosting” of graduate students Princeton(Schowalter) and University of

Wisconsin (Bob Bird) The advice of Bob Dibble

(Mech E. UC Berkeley)

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The two color mimeo

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Other people

Arthur Lodge John Ferry Don Baird (and the watch)

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Here was the Bird/ERC gang

Albert Co Univ Maine

Alan Graham Los Alamos Roger Grimm

Amaco/Livermore

Richard Christianson Colo School Mines

Moshe Gottlieb BenGurion Univ.

35# Hubbard Squash

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Bob’s visit to Princeton l991, and the hike

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Then there were languages

Bob wrote out my kid’s names in Japanese when he visited Princeton in l991

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Thanks for the memories

Jean Lippert – who ran the zoo

Thanks for the mentoring, memories, and being a part of the Wisconsin tradition !

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Facile Production of Multifunctional Nanoparticles for Difficult to

Deliver Therapeutics

Robert K. Prud’homme, Lawrence Mayer1 , Kim Chan2, Prabhas Moghe3, Debrah

Laskin3, Patrick Sinko3, Zoltan Szekely3, Vasanthi Sunil3, Leo Einck4, Venkata Reddy4,

Michael Natchus5, Randy Howard5, Dennis Liotta5

Brian Johnson, Walid Saad, Ying Liu, Marian Gindy, Stephanie Budijono, Margarita Herrera- Alonso, Varun Kumar, Suzanne D’Addio, Carlos Figueroa, Nathalie Pinkerton, Adam York

Dept. Chemical Engineering Princeton University

Celator, U. Sydney, Rutgers, Sequella, Emory

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Bob Bird’s academic lineage

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Ruben Savergold’s enlistment from Madison

21 March 1864

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When he taught it made things look simple

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He gave us freedom in defining our project

I was interested in polymer foams. Bob suggested I write to academics and industrial researchers to get some input. Here is Howard Brenner’s reply:

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Next Generation Nanoparticles (NPs) Motivation

• Bioavailability: 40% of new drug compounds are hydrophobic

• EPR targeting of solid tumors • Targeting toxic API • Multiple drugs cocktails • Imaging where toxic APIs go • Aerosol drug delivery • siRNA and peptide delivery More than just small

•Size •Surface functionality •Stoichiometric encapsulation of multiple species: cocktails

•Imaging plus delivery •Targeting plus delivery

•Scaleable and manufacturable

Tumor vasculature

Nanoparticles

11 µm

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Next Generation Nanoparticles (NPs)

Tumor vasculature

Nanoparticles

11 µm

“…over the next few years some of the complex theranostic strategies published rampantly in chemistry journals will fall out of contention…For me, something that’s too difficult to make or too complex to sustain in large-scale production is not what we are interested in.”

J.Janijic C&EN Sept 26,2011.

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Outline

1. Why nanoparticles?- applications 2. How nanoparticles– impinging jet mixing

and Flash Nano Precipitation a. Mixing b. Block copolymer micellization c. Control of particle size

3. Controlled release from nanoparticles 4. Multi-functional nanoparticles -- imaging 5. Targeting with surface functionalized

nanoparticles

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Block copolymer directed rapid precipitation

Nanoparticle formation by Flash NanoPrecipitation

Non solvent Copolymer Solute Solvent

Dead Micelle

PROTECTED NANO

PARTICLE

tmix

Micellization tagg

tng

Nucleation and Growth

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Flash Precipitation Image analysis = 249 nm Light diffraction = 370 nm SSA = 22 m2/gram

“Typical” Precipitation Image analysis > 10 um Light diffraction > 10 um SSA > 0.6 m2/gram

10 µm TEM’s by Helmut Auweter (BASF)

Polymer Protected vs Unprotected Particle Growth

1 µm

200 nm

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Particle Diameter (µm)0.01 0.1 1 10

Volu

me%

in R

ange

0

2

4

6

8

10

12

14

16

18

20

Tota

l Vol

ume%

0

20

40

60

80

100

Volume Mean D[4,3] = 0.45 µm

Standard Surface Area S.S.A.= 15.4 m^2/gram

% in range % less than size

Particle Diameter (µm)0.01 0.1 1 10

Inte

nsity

% in

Ran

ge

0

2

4

6

8

10

Tota

l Int

ensi

ty%

0

20

40

60

80

100

2nd Order Cumulant D[6,5] = 0.088 µm

% in range % less than size

Change Inputs to Process

2.8 m/s, (2.6 wt% drug, 0.4 wt% Copolymer)

H20

PROTECTED NANOPARTICLE

THF PS-b-PEO β-carotene

τmix τagg

τng

Size

% in

Ran

ge

Si

ze%

in R

ange

Cum

ulat

ive

%

Cum

ulat

ive

%

“FLASH” Nanoparticles Precipitation Size Control

D= 450 nm

D= 88 nm

•High throughput, > 2 wt% active

•High NP loading

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Active Compounds for Encapsulation

β-carotene

Insoluble

Estradiol (hormone)

Vitamin E

Paclitaxel (anti-cancer agent)

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Block-copolymers used for nanoparticle formation

Poly (ethylene glycol)-b-poly (styrene)

Poly (acrylic acid)-b-poly (butyl acrylate)

Poly (ethylene glycol)-b-poly (caprolactone)

Poly (ethylene glycol)-b-poly (lactic acid)

3k-b-1k HO

O

PSn

PEOm

5k-b-4.2k

5k-b-2.9k

7.59k-b-7.56k

OO

H2C

CH3

O

OH

PAA PBA

3

mn

O

OO

mmPEG PCL

n

O

mPEG PLAn

OO

O CH3

O CH3m

Hydrophilic block Hydrophobic block

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Syringe Pump Mixing Heads: d= 0.25 to 1 mm ∆ = 4.8 to 19

∆ = I/d

Z

H

d

I = D

H = 0.8DZ = 1.2D

δ

CONFINED IMPINGING JETS (CIJ) MIXER

Scaleup: • 3mg lab samples • lab system produces 3.5 kg/day at 17 psi •Scales Q~ ∆P⋅R3

•∆P → 100 psi •R =250 µm → 1 mm: Q= 64X

•Scales to 1,000 kg/day

Johnson, et al. , AIChE J, 49,2264 (2003) Liu et al. ,AIChE J 52 731-744 (2006)

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POWER DENSITY

mVP3ρ

ε =

Mixing Energy Density d

1 2

3

∆ = I/d = 4.76 I

POWER - Isothermal mechanical energy balance (BSL 1960)

+=

2

1113

12

18 m

mudPρπ

3x

2d dIV yym ∆=∝

πMESOMIXING VOLUME - Define via dimensional analysis

+

∆=

2

113

13

141

mm

dyu

ρ

ρε

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TURBULENCE THEORY - Mixing Two Fluids

5.12

Re4

−∝==Ddiffusionmix

λττ

A B

Molecular Diffusion

4/13

=

ενλ

Mixing Time

Kolmogorov Length Scale

B. Johnson, R.Prud’homme AIChE J (2003)

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(FAST) (SLOW)

Competitive Reactions as a “Chemical Ruler”

D A

Products B Eq 1.00

A Eq 1.05

→ =

+ 8 10 1 k

Products

B D

Eq 1.00 →

< +

10 2 k

Competitive(Parallel) Reactions

Characteristic Reaction Time

A

B A

B D

D

rxnmix ττ ≥ rxnmix ττ

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0.001

0.010

0.100

1.000

10 100 1000 10000

317 mSec - Dimethoxypropane181 mSec61 mSec28 mSec16.7 mSec9.5 mSec6.5 mSec4.8 mSec343 mSec - Ethylchloroacetate0.04 breakpointsSlope = -3/2

500A-Y2Xd = 0.50 mm∆ = 4.76δ = 2d

Bor Ck2

1=τ

Equivalent Mixing Time at X = 0.04

X =

Con

vers

ion

of S

low

Reac

tion

(DM

P)

Jet Reynolds Number

Selectivity vs Reynolds Number

CBo = 65 mol/m3

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0.001

0.010

0.100

1.000

0.01 0.1 1 10 100 1000 10000

317 mSec-DMP Hydrolysis181 mSec61 mSec28 mSec16.7 mSec9.5 mSec343 mSec - EthylchloroacetateSlope=1

Onset of "Turbulent Like"

Motion atRe = 90

500A-Y2Xd = 0.50 mm∆ = 4.76δ = 2d

X =

Con

vers

ion

of S

low

Rea

ctio

n

constantx Re 2/3

2 Bo

rxn

mix CkDa ==ττ

Bor Ck2

1=τ

Selectivity vs. Damkohler Number: X ∝ Re-3/2

X = 0.04

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Competitive Micellization and Precipitation Kinetics

Micellization τagg

Stabilization

τng Nucleation

and Growth

Non solvent Copolymer Solute Solvent

Dead Micelle

PROTECTED NANOPARTICLE

τmix

Process: Competitive Time Scales

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• Control super -saturation by changing solvent quality or solute concentration.

• Higher super saturation leads to smaller particles

Control of particle size

0 100 200 300 400 500 60050

100

150

200

250

300 1:1 H2O:THF 2:1 H2O:THF 3:1 H2O:THF 5:1 H2O:THF

Parti

cle D

iam

eter

(nm

)

Average Re

1. Mixing intensity

2. Super- saturation

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Outline

1. Why nanoparticles?- applications 2. How nanoparticles– impinging jet mixing

and Flash Nano Precipitation a. Mixing b. Block copolymer micellization c. Control of particle size

3. Controlled release from nanoparticles 4. Multi-functional nanoparticles -- imaging 5. Targeting with surface functionalized

nanoparticles

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NO prodrugs: anti cancer and inflamation

With Larry Keefer (NIH): diaziniumdiolates Glutathione releases NO from prodrug

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NO prodrugs: anti cancer and inflamation

With Larry Keefer (NIH): diaziniumdiolates Glutathione releases NO from prodrug

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siRNA for gene silencing

•siRNA complex cleaves mRNA and stops protein expression

•Problem is delivery

+

buffer siRNA

ethanol cationic lipid

structural lipid

+

+ + +

+ + +

+

+

•Successful prep of 100 nm PEG protected siRNA

•Stable and transfect efficiently

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Secondary crystallization, Ostwald ripening

Drug Instability

Drug + Hydrophobic “anchor” - Increased drug hydrophobicity - Protect from enzymatic hydrolysis - Decreased partitioning from the nanoparticle - Release dependent on prodrug cleavage kinetics (water concentration in the particle core) - Allows “cassette” formulation of mixed drug formulations

Drug Linker Hydrophobe

time

Stable Drug Nanoparticles by Conjugation as Prodrugs

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Drug OH

Drug O SiRR

RDrug O

O

R

R

Drug OO

R

C

O

NH

HC

HC

OH

C

O

O

H3C

HOO

O

O

CH3OHOOC

O

H3C

CH3

C OC CH3O

HCH3

HOH OH

OOH

NH

CH3 CH3 CH3

CH3

O

O

CH3H3CO

OO

H3C

OCH3

OH

OH

N

H

N

NCH3

OHCH3

HO

Paclitaxel Rifampicin

Estradiol

R = hydrophobic molecule

Hydroxyl Protecting Groups

Prodrug Chemistry

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Paclitaxel conjugate release rate

Ansel,Mayer,Saad, Prud’homme J Med. Chem (2008)

Conjugation of drug to paclitaxel

Measure circulating H3 paclitaxel in Foxn1nu mice serum

Conjugating hydrophobic “anchors”

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Outline

1. Why nanoparticles?- applications 2. How nanoparticles– impinging jet mixing

and Flash Nano Precipitation a. Mixing b. Block copolymer micellization c. Control of particle size

3. Controlled release from nanoparticles 4. Multi-functional nanoparticles -- imaging 5. Targeting with surface functionalized

nanoparticles

PrincetonUniversity

Coumarin Nanoparticles for Imaging

O

CH3

CH3

CH3 CH3 CH3

OCH3

CH3

CH3

CH3

O O

NHO

THF (12ml/min)

H2O (120ml/min)

O O

O O

O

O H O

n m

Coumarin-VitE

mPEG(5k)-b-PCL(6k) Uptake of 80nm LHRH-PEG protected nanoparticles into MS578T breast cancer cells.

Akbulut et al. Adv. Fn Mat. (2009)

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X-ray CT Imaging

Accounts for 80% of all diagnostic imaging

High absorption coefficient; 2.7x greater contrast than iodine

Reduced bone interference & enhanced soft tissue absorption

Prolonged circulation Biological safety Combined therapy and imaging

Gold nanoparticles as X-ray contrast agents

Block copolymer stabilized

gold nanoparticles

5 nm

Transmission Electron Microscopy

Au

DIW

DIW

THF THF

PEG-b-PCL

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Composite Rifampicin NPs

Simultaneous Organic/Inorganic Active Loading

Au CNPs (a)

Rifampicin + Au CNPs (b)

C1 2- Au 0. 01 6 wt%

0. 01 6 wt%

Rifampicin 0 wt% 0. 4 wt% PEG- b- PCL 0. 4 wt% 0. 4 wt% Diameter ~95 nm ~300 nm

OH OH

OOH

NH

CH3 CH3 CH3

CH3

O

O

CH3H3CO

OO

H3C

OCH3

OH

OH

N

H

N

NCH3

5 nm

a

with Y. Liu, PhD Thesis, Princeton 2007

100 nm

b

Anti-tuberculosis drug

[1] S. Torquato et al. Phys. Rev. Lett. 2000, 84, 2064-2067

•X-ray CT Imaging •Multifunctional imaging and delivery

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Magnetic CNPs for MRI Contrast Enhancement

Clinical MR Image

T2 relaxivity (r2) comparable to current FDA approved iron oxide MRI contrast agents

Additionally demonstrated co-encapsulation of drugs without adverse effect on measured relaxivity

Feridex ®

15 nm cobalt-ferrite colloids (C. Rinaldi)

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Outline

1. Why nanoparticles?- applications 2. How nanoparticles– impinging jet mixing

and Flash Nano Precipitation a. Mixing b. Block copolymer micellization c. Control of particle size

3. Controlled release from nanoparticles 4. Multi-functional nanoparticles -- imaging 5. Targeting with surface functionalized

nanoparticles

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Active targeting

Over-expression of specific receptors at surfaces of diseased cells

Allows for targeting of metastatic tumor cells, macrophages, inflamation, etc.

NP surfaces functionalized with ligands Small molecules (folate, mannose) Peptides and proteins (LHRH, VEGF) Antibodies (2C5)

Key research questions Density of ligand attachment Preservation of ligand integrity

(functionality) PEG Mw Presentation of PEG vs steric layer

Nature Reviews 2006, 5, 147-159

Gindy, Biomaterials (2008) Akbulut, Adv. Fn. Mat. (2009)

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ML3.9 scFv Anti-HER2 (ErbB2) 1 x 10-9 M affinity

A5 scFv Anti-HER3 (ErbB3) 1.6 x 10-7 M affinity

VL

VL

VH

VH

Renal Cutoff ~65kDa

Nanoparticle targeting sweet spot: Cost 10-3 of antibodies Kynamro (Genzyme) $176,000/yr Can surface presentation on NPs increase binding avidity? One NP carries 200,000 drug molecules

Targeting alternatives to antibodies

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Variable domain antibody targeting

9

ELN

– N

anop

artic

les 2

013

- 000

48

Blank Cells

W26A-NPs (300 ug/mL)

•Centyrin scaffold 10K (Janssen Pharma)

•Pre functionalize BCP •Assemble NP

Control (PEG NP)

5% ligand targeted

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Targeting of Macrophages for TB

1. Engineer a range of different surface-coated and sized NPs

2. Explore the effects of temperature, incubation time, NP size, surface coating properties on uptake by macrophages

3. Coat NPs with ligands to enhance NP uptake by MACs

Nucleus

Surface Adherence

Receptor Mediated Endocytosis

Phagocytosis

Size?

Surface Coating?

Temp? Time?

Note: also doing folate targeting (U Mich, click chemistry), VEGF (Sibtech, maleimide), antibody (V. Torchilin, pNP), LHRH

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Drugs: • INH – current frontline drug

• SQ641 – novel anti-TB drug (Reddy et al. Antimicrob. Agents Chemother. 2008)

•Cyclosporine A (CsA) - Pgp efflux pump blocker

•Protocol: • Mouse macrophages were infected with MTB

• 1 day incubation at 2 x MIC • RLU determined on Day 7

In Vitro anti-TB efficacy

Dual drug delivery from nanoparticles

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OHOHO

OHOH

ON3

OAcOAcO

OAcOAc

OAc69 %

OO

m nOOH

m n NaH

THF+ Br

53 %

+CuBr, TBTA

THF

OHOHO

OHOH

ON

N N

O PEG b PS

24 %

2

3

1

Chernyak, A. Y.; Sharma, G. V. M.; Kononov, L. O.; et al. Carbohydr. Res. 1992, 223, 303. Sanki, A. K., Mahal, L. K. Synlett 2006, 3, 455. Geng, J.; Mantovani, G.; Tao, L. J. Am. Chem. Soc. 2007, 129, 15163.

Mannose targeting of macrophages for TB

III. Click Chemistry Conjugation

• Mannose –azide and PS-b-PEG-alkyne • Preform polymer and mix neutral and functional block copolymers

during NP formation. Vary Mw PEG, ratio of neutral/functional, end groups of neutral polymer

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Mannose Receptor (MR) Targeting

Binding and uptake of NPs is enhanced 3-fold when the corona is composed of 9% Mannose terminated

ends.

Gindy, Biomaterials (2008) Akbulut, Adv. Fn. Mat. (2009)

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Outline

1. Why nanoparticles?- applications 2. How nanoparticles– impinging jet mixing

and Flash Nano Precipitation a. Mixing b. Block copolymer micellization c. Control of particle size

3. Controlled release from nanoparticles 4. Multi-functional nanoparticles -- imaging 5. Targeting with surface functionalized

nanoparticles

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Cassette Nanoparticle Systems

Novel approach to formation of multifunctional , multicomponent nanoparticle formulations

Stabilizing

Targeting

Multiple drugs, proteins, polypeptides

Imaging

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Thanks for the memories

Jean Lippert – who ran the zoo

Thanks for the mentoring, memories, and being a part of the Wisconsin tradition !

bungee jumping at Kawarau Bridge, NZ

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Aerosol Drug Delivery with Nanoparticles

Load Dave Edward’s (Harvard) Aeroparticles with Nanoparticles

Large solid particles are

inertially deposited in the

mouth and throat

d>3µm

Small particles cannot be

captured by filtration d

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Integrated NanoParticle Process

Non solvent Copolymer Solute Solvent

τmix

Water, Sugar

Air

Prud’homme

NanoPrecipitation

Edwards

PNP Spray Drying

•BASF β-carotene business built on impingement jet process, 1400 kg/day

•Alkemes aerosol drug delivery business built on PNP spraying

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Spray drying PNAP particles

Four Stream Vortex Mixer

Spray drier (Niro Atomizer Portable Spray drier)

Spray dry a mixed stream of nanoparticles and sugar/leucine

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Rifampicin NPs redispersion

• Redissolution of PNAPs spray dried with 50% rifampicin, 50% leucine

• 87% rifampicin recovered in NP form after dissolution

• NP size: initial 170 nm • After spray drying and

dissolution 145 nm

Initial NP

After redispersion

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Particle Morphology

1 µm

Chew and Chan. Pharmaceutical Research. 1999

10 µm

Spray Drying Spray Freeze Drying

Size depends on Solute diffusion

vs Solvent drying

100⁰C

Size is fixed Solvent freezing

“traps” solute -196⁰C

Maa et al. 199

PNAPS for Aerosol Drug Delivery

Spray Drying

LN2

Spray Freeze Drying

1% 5% 2% 50%

BSA

Mannitol

Lysozyme

•Size independent of conc. •Density controlled by conc.

•50% nanoparticle w/ leucine matrix

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Aerosol Performance w/ Spray Freeze Drying

ga dd ρ=

dg = 20 ± 8 µm

Throat

Lungs

• dg is fixed in spray freeze drying • FPF is optimized through particle

density

Mannitol Lysozyme BSA

Cascade aerosol impactor Fine particle fraction: d< 3μm

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Outline

1. Why nanoparticles?- applications 2. How nanoparticles– impinging jet mixing

and Flash Nano Precipitation a. Mixing b. Block copolymer micellization c. Control of particle size

3. Controlled release from nanoparticles 4. Multi-functional nanoparticles -- imaging 5. Targeting with surface functionalized

nanoparticles (TB) 6. Post Processing and Aerosol delivery

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Activity is equivalent for NP form Cell types: •human leukemia U937 •human non-small cell lung cancer H1703

Equivalent IC50 for PEG protected NPs as that of naked drug compounds

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Amphotericin B Nanoparticles

Amphotericin antibiotic

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Outline

1. Why nanoparticles?- applications 2. How nanoparticles– impinging jet mixing

and Flash Nano Precipitation a. Mixing b. Block copolymer micellization c. Control of particle size

3. Controlled release from nanoparticles 4. Multi-functional nanoparticles -- imaging 5. Targeting with surface functionalized

nanoparticles (TB) 6. Aerosol delivery

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Coumarin Nanoparticles for Imaging

O

CH3

CH3

CH3 CH3 CH3

OCH3

CH3

CH3

CH3

O O

NHO

THF (12ml/min)

H2O (120ml/min)

O O

O O

O

O H O

n m

Coumarin-VitE

mPEG(5k)-b-PCL(6k) Uptake of 80nm PEG protected nanoparticles into MS578T breast cancer cells.

Akbulut et al. Adv. Fn Mat. (2009)

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Rigid Beads 3-10μm

Targeting to the Lung depends on size and deformability of microparticles (Pat Sinko – Rutgers)

Too Small for Accumulation

Too Large for Clearance

Chao, P.Y. et al. Anti-Cancer Drugs 2010, 21, 65

Soft Gels – 55μm 1 hour

4 days 7 days

12 hrs

Lungs kidneys

From body

To body Venous Filtration Pathway

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Heirarchical targeting and delivery

Microparticle -Scale Targeting to Lungs • Targeting to lungs using size

and modulus • RBC mimics • “Gel Microparticles” GMPs • Degradable PEG gels

0.2-2 µm

Nanoparticle -Scale Targeting to Tumor

• Targeting to tumor using triggered release of PEG gel network

• Targeting to tumor using release kinetics of pro-drug conjugates from nanoparticles

GMP

100 nm

NP microfluidics emulsification

8-40 µm

Nanoparticles Proteins

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Formation degradable GMPs

Aqueous Phase: Polymer, Nanoparticles, Crosslinking Agent

Oil Phase

Crosslinking

Stable GMPs

Anna S.L. et al. Applied Physical Letters 2003, 82, 364

Microfluidics Chemistry – Modulus and Degradation

PLA-b-PEG-b-PLA

degradable link

degradable link

Photopolymerize microfluidic drops into microgel particles

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Gel Degradation Kinetics and NP Release

3.4KL5 Hydrogel a Lysozyme b BSA c Dextran

Lu and Anseth, Macromolecules, 2000, 33, 2509-2515 PEG 400 PEG 600

UV Exposure Time

•Photopolymerize microgels • (compound concentration, wavelength)

•Gel mesh size ~ modulus

Coming to MadisonThe two color mimeoOther peopleHere was the Bird/ERC gangBob’s visit to Princeton l991, and the hikeThen there were languagesThanks for the memoriesSlide Number 14Slide Number 15Slide Number 16Slide Number 17Facile Production of Multifunctional Nanoparticles for Difficult to Deliver TherapeuticsBob Bird’s academic lineageRuben Savergold’s enlistment from MadisonWhen he taught it made things look simpleHe gave us freedom in defining our projectNext Generation Nanoparticles (NPs)Next Generation Nanoparticles (NPs)OutlineNanoparticle formation by Flash NanoPrecipitationPolymer Protected vs Unprotected Particle Growth“FLASH” Nanoparticles Precipitation Size ControlSlide Number 34Block-copolymers used for nanoparticle formationCONFINED IMPINGING JETS (CIJ) MIXERSlide Number 39Slide Number 40Slide Number 41Slide Number 42Slide Number 43Process: Competitive Time ScalesControl of particle sizeOutlineNO prodrugs: anti cancer and inflamationNO prodrugs: anti cancer and inflamationSlide Number 68Slide Number 69Slide Number 70Paclitaxel conjugate release rateOutlineSlide Number 74Slide Number 77Slide Number 78Slide Number 80OutlineActive targetingSlide Number 88Variable domain antibody targetingTargeting of Macrophages for TBIn Vitro anti-TB efficacySlide Number 94Mannose Receptor (MR) TargetingOutlineCassette Nanoparticle SystemsThanks for the memoriesSlide Number 102Slide Number 103Aerosol Drug Delivery with NanoparticlesIntegrated NanoParticle ProcessSlide Number 106Rifampicin NPs redispersionParticle MorphologyPNAPS for Aerosol Drug DeliveryAerosol Performance w/ Spray Freeze DryingOutlineSlide Number 112Slide Number 113Amphotericin B Nanoparticles�OutlineSlide Number 117Targeting to the Lung depends on size and deformability of microparticles (Pat Sinko – Rutgers)Heirarchical targeting and deliveryFormation degradable GMPsGel Degradation Kinetics and NP Release