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BTM Nanocapsules for Formulationof Drugs and Vaccines and Imaging Agents

Carolina Seeds of InnovationNovember 4, 2010

Russell J. Mumper, Ph.D.Center for Nanotechnology in Drug Delivery

Division of Molecular PharmaceuticsUNC Eshelman School of Pharmacy

University of North Carolina at Chapel HillChapel Hill, North Carolina

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Nanotemplate Engineering

Focus Areas

Materials (Handbook of Pharmaceutical Excipients)

Engineering & Characterization

Cell Interaction / Uptake

Biofate & Biometabolism

Bio- and Hemocompatibility (toxicological aspects of NPs)

Cell and Tissue Targeting (therapeutics)

Therapeutic (and Imaging) Areas

– Addressing resistance in human cancer using nanotechnology – Subunit (protein) vaccines for HIV

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OilDrug (Rx)

Surfactant(s)

Oil

Surfactant(s) Water

Clear, Stable Oil-in-Water

Microemulsion“Nanotemplate”

at 50-65oC

+ Water

Cool to 25oC

Enables manufacturing of stable NPs <200 nm using a reproducible and scalable process Manufacturing process is as few as 3-steps and is completed in one vessel Overcomes the limitations of commonly used methods to make sub-micron sized particles

Step 3Step 2Step 1

Add, Heat & Mix at 50-65oC

Nanoparticles or Nanocapsules

Nanotemplate Engineering

oil droplets

+_

Ni

OH

PEG-SH-NH2

-COOH

RXN

Y

Ligand

HO

RxPEG Rx

RxRx

Rx

Rx

Rx

Rx

Rx Rx

oil droplet nanotemplate

= surfactant

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Vitamin E TPGS(d-Alpha Tocopheryl Polyethylene Glycol 1000 Succinate)

CH3 (CH2)17 (OCH2CH2)20OHBrij 78(Polyoxyethylene 20 stearyl ether)

E78 Nanoparticles vs. BTM Nanocapsules

Cetyl Alcohol (m.p. 49oC)

CH3 (CH2)14CH2OH

Miglyol® 812 Caprylic/Capric TriglycerideC8 (50-65%); C10 (30-45%)Oil Phase

Surfactants

+

Solid Lipid E78 Nanoparticles Oil-Filled BTM Nanocapsules

E78 NPs

BTM NPs

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A New Nanocapsule Formulation - “BTM NPs”

First Generation BTM NPs identified by Sequential Simplex Optimization

Composition: liquid tri-glyceride core with two surfactants Scalable, one vessel manufacturing process Lipid/drug ~ 20-30:1 w/w; [drug] up to 1.5 mg/ml Sustained-release of drug Can be pegylated to make ‘pegBTM NPs’ Easily sterile filtered Can be lyophilized with no cryoprotectant Very stable in suspension or lyophilized ‘Plug & Play’ platform based on oil properties Very well tolerated, repeated i.v. injection up to 750 mg/kg

Dong et al. Eur. J. Pharm. Biopharm. (2009)

Lyophilized BTM NPs

15 s after rehydration

6Mice (n=6/group) were dosed i.v. with PX (4.5 mg/kg) on day 0, 7, 14, and 21

0

200

400

600

800

1000

1200

0 5 10 15 20 25 30

Day

Tum

or

siz

e (

mm

3 )

* * * * * * *

#

Saline

Empty BTM NPs (4.5 mg/kg)

Taxol (4.5 mg/kg)

Taxol (20 mg/kg)

Empty BTM NPs + Taxol (4.5 mg/kg)

PX BTM NPs (4.5 mg/kg)

In-Vivo Efficacy Study in Nude Mice Bearing P-gp+ Resistant Human Ovarian Tumors

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Retreatment of Mice

0

100

200

300

400

500

600

700

800

900

1000

0 3 6 9 12 15 18 21 24 27 30

0

200

400

600

800

1000

0 1 2 3 4 5 6 7 8 9 10 11

542

303

519

364 340

509559

420383**

***

*

Taxol-failed mice can be salvagedwith PX BTM NP treatment

Previously PX BTM NP treated mice respond to new course of PX

BTM NP treatment

Day Day

4.5 mg/kg

7.5 mg/kg

PX BTM dose

Tum

or

siz

e (

mm

3 )

Day 31 ofStudy #2

Day 49 ofStudy #2

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Enhancement of Tumor MRI Image with BTM-DTPA-Gd NPs

5 hr after i.v. injection in A549 s.c. xenograft tumors ~50-70 mm3

Compliments of Dr. Michael Jay in collaboration with SAICF at UNC-BRIC using 9.4T Micro-MRI

Control BTM-DTPA-Gd NPs

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J. Biomed. Nanotech. (2007) Pharmaceutical Research

(2007) Vaccine (2004, 2006)

HIV/AIDS (2009)

Nano-based Subunit HIV Vaccines

Dendritic Cell

Toll-like Receptor (TLR-9)

ReceptorMHC I

MHC II

DC targetingLigand

PEG

Tat (1-72)

Gag p24

Adjuvant (PRL)

A DC-targeted nanoparticle with conserved proteins Tat (1-72) and Gag p24 to generate protective Th1, CTL, and neutralizing antibody responses that may be further enhanced by co-

delivery of Adjuvants (PRLs)

Tat & Gag antigens: conserved; critical; CTLs detected in LTNPs

NIH-NIAID R01 AI058842

NP benefits: Increased DC uptake/processing/

presentation Dose sparing Enhance MHC1 processing Enhance Th1-type responses Enhance (neutralizing) antibodies Co-delivery of antigen/adjuvant

DiOC18 NPs in DC

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Alum(1 µg)

Naive0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0P

41

sp

ec

ific

Ig

GO

.D.

at

45

0 n

m

1,000

2,000

5,000

10,000

E78-Ni(1 µg)

BTM-Ni (1 µg)

BALB/c mice (n=8/group) were dosed by s.c. injection on day 0 and 14; ELISA day 28 Doses: 0.1, 0.5, or 1 mg p41 His-tag p41 provided by Dr. Robert Seder, NIH-NIAID Vaccine Research Center

p41 Immunization; BTM-Ni vs. E78-Ni NPs

BTM-Ni(0.5 µg)

*

BTM-Ni(0.1 µg)

#

*

*

*#

#

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Concluding Remarks

“Nanoparticle Compositions Comprising Liquid Oil Cores” (PCT /US2009/060593 )

“Translational Nanotechnology” – all required elements

Nanotemplate Engineering

simple, one-vessel process, reproducible, scalable, cost-effective keys: 1) physical chemistry/pharmacy 2) excipient selection

Some ‘GRAS’ or USP/NF excipients may be ‘biological modifiers’

Nano-based Drug Delivery Systems

Improve drug solubility / bioavailability Address MDR in cancer to improve outcomes Can be used for imaging Co-delivery of antigens / adjuvants for improved vaccines

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Russell J. Mumper, Ph.D.Center for Nanotechnology in Drug Delivery

Division of Molecular PharmaceuticsUNC Eshelman School of Pharmacy

University of North Carolina at Chapel HillChapel Hill, North Carolina

E-mail: mumper@email.unc.edu