BTM NPs - Research at Carolina

Report
BTM Nanocapsules for Formulation
of Drugs and Vaccines and Imaging Agents
Russell J. Mumper, Ph.D.
Center for Nanotechnology in Drug Delivery
Division of Molecular Pharmaceutics
UNC Eshelman School of Pharmacy
University of North Carolina at Chapel Hill
Chapel Hill, North Carolina
Carolina Seeds of Innovation
November 4, 2010
1
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


2
Nanotemplate Engineering
 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
Oil
Add, Heat & Mix
at 50-65oC
Surfactant(s)
Oil
Drug (Rx)
Surfactant(s)
Water
+ Water
Step 2
at 50-65oC
Step 1
Y
oil droplet
nanotemplate
Ligand
PEG
+
Clear, Stable
Oil-in-Water
Microemulsion
“Nanotemplate”
Rx Rx
Rx
Rx Rx
Rx
Rx Rx
Rx Rx
RXN
-SH
-NH2
-COOH
Step 3
= surfactant
_
HO
Cool to 25oC
oil droplets
Ni
Nanoparticles
or Nanocapsules
PEG
OH
3
E78 Nanoparticles vs. BTM Nanocapsules
Solid Lipid E78 Nanoparticles
Cetyl Alcohol
(m.p. 49oC)
Oil Phase
Oil-Filled BTM Nanocapsules
Miglyol® 812
Caprylic/Capric Triglyceride
C8 (50-65%); C10 (30-45%)
CH3 (CH2)14CH2OH
E78 NPs
BTM NPs
+
Surfactants
Vitamin E TPGS
(d-Alpha Tocopheryl Polyethylene
Glycol 1000 Succinate)
Brij 78
(Polyoxyethylene 20 stearyl ether)
CH3 (CH2)17 (OCH2CH2)20OH
4
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
Lyophilized BTM NPs
15 s after
rehydration
Dong et al. Eur. J. Pharm. Biopharm. (2009)
5
In-Vivo Efficacy Study in Nude Mice Bearing
P-gp+ Resistant Human Ovarian Tumors
Saline
Empty BTM NPs (4.5 mg/kg)
Taxol (4.5 mg/kg)
1200
Taxol (20 mg/kg)
Empty BTM NPs + Taxol (4.5 mg/kg)
Tumor size (mm3)
1000
PX BTM NPs (4.5 mg/kg)
800
600
#
400
200
*
*
*
*
*
*
*
0
0
5
10
15
20
25
30
Day
Mice (n=6/group) were dosed i.v. with PX (4.5 mg/kg) on day 0, 7, 14, and 21
6
Retreatment of Mice
Taxol-failed mice can be salvaged
with PX BTM NP treatment
Previously PX BTM NP treated
mice respond to new course of
PX BTM NP treatment
1000
1000
900
Tumor size (mm3)
800
800
700
600
600
542
500
559
519
400
*
300
*
*
364
303
200
*
509
400
*
*
420
383
340
200
100
0
0
0
Day 31 of
Study #2
3
6
9
12
15
18
21
24
27
30
Day
PX BTM dose
0
1
Day 49 of
Study #2
2
3
4
5
6
7
8
9
10 11
Day
4.5 mg/kg
7.5 mg/kg
7
Enhancement of Tumor MRI Image with
BTM-DTPA-Gd NPs
Control
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
8
NIH-NIAID R01 AI058842
Nano-based Subunit HIV Vaccines
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)
DiOC18 NPs in DC
NP benefits:
Increased DC uptake/processing/
presentation
Dose sparing
Enhance MHC1 processing
Enhance Th1-type responses
Enhance (neutralizing) antibodies
Co-delivery of antigen/adjuvant
Dendritic Cell
Toll-like Receptor
(TLR-9)
Tat (1-72)
MHC I
Receptor
PEG
Adjuvant
(PRL)
DC targeting
Ligand
J. Biomed. Nanotech. (2007)
Pharmaceutical Research (2007)
Vaccine (2004, 2006)
HIV/AIDS (2009)
Gag p24
Tat & Gag antigens: conserved; critical; CTLs detected in LTNPs
9
p41 Immunization; BTM-Ni vs. E78-Ni NPs
4.0
*
3.5
1,000
2,000
P41 specific IgG
O.D. at 450 nm
3.0
5,000
2.5
10,000
*
2.0
1.5
*
*
1.0
#
#
0.5
#
0.0
BTM-Ni
(0.1 µg)
BTM-Ni
(0.5 µg)
BTM-Ni
(1 µg)
E78-Ni
(1 µg)
Alum
(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
Naive
10
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
11
Russell J. Mumper, Ph.D.
Center for Nanotechnology in Drug Delivery
Division of Molecular Pharmaceutics
UNC Eshelman School of Pharmacy
University of North Carolina at Chapel Hill
Chapel Hill, North Carolina
E-mail: [email protected]
12

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