Download Normal tissues - 16th International Congress on Neutron Capture

Development of Boron Containing Nanodiamonds
for Boron Neutron Capture Therapy
Ming-Hua Hsu, Ph. D.
Nuclear Science and Technology Development Center, NTHU
June 16, 2014
@ ICNCT 16, Helsinki, Funland
Outline
1. Nanomedicine
2. Boron Containing Polmeric nanoparticle for
BNCT
 PLA-PEOz-B-pin
 PLA-PEOz-B-cage
3. Boron Containing Nanodiamonds for BNCT
4. Boron Nanoparticles for BNCT
2
Nanomedicine: Nanotechnology to Medicine
Apply Nanotechnology:
•Maintain drug activity.
• Increase water solubility.
Nanomedicine
Use of nano-materials in
medicine  unique
medical effects
• Delivery drug by nanoparticle
• Increase drug efficiency
• Traceable or directable drug
The Advantage of Nanoparticles
• High surface/volume ratio and good solubility
• Nanometer size lead to faster movement and easy entry into cell
• Nanoparticle vector can penetrate endothelial barriers to reach tumor
sites
drug
nanoparticles
The enhanced permeation and retention (EPR) effect
Nature Reviews Cancer 12, 39-50 (January
2012)
Micelle-based Drug Delivery System
The micelle must:
(i) be small enough (∼10—200 nm) to
effectively penetrate into tissue;
(ii) be unrecognizable by the mononuclear
phagocyte system (MPS) for a sufficient
time to allow accumulation in target tissue;
(iii) be eliminated from the organism either
after degradation or dissolution;
(iv) locate and interact with the target cells;
(v) have tunable stability;
(vi) improve the pharmacokinetic (PK)
profile of the encapsulated drug cargo;
(vii) possess high loading capacity; and
(viii) be synthesized in a reproducible,
facile method which is reasonably
inexpensive
Development of BNCT Drug Deliver System Based
on Polymeric Material PLA-PEOz
Polylactide, PLA
•
•
•
•
•
•
Biocompatible
Biodegradable
Low immunogenicity
Good mechanical properties
FDA-approved for clinical use
Hydrophobic
Polyoxazoline
•
•
•
•
•
•
Pseudopolypeptides
Adjustable
Biocompatible
Low immunogenicity
Stealth
FDA-approved as a food contact
agent
• Hydrophilic
Strategy of Copolymers Synthesis
Synthetic Scheme of Bpin-PLA-PEOz
Characterizations of synthesized polymers
Table1 Characterizations of synthesized polymers
polymer
Mna
Polydispersity
index (PDI)b
Yield (%)
Bpin-PLA
7218
1.21
98
Bpin-PLA-PEOz
14247
1.24
62
a Estimated
by 1H NMR . b Estimated by GPC.
CMC (wt %)
7×10-4
Boron Content of Bpin-PLA-PEOz micelle
Synthesis of
Phenylboronic acid derivative
(PBAD)
Bpin-PLA-PEOz
Bpin-PLAPEOz/PBAD
• 6.06 ± 0.3 μg B/ml
• 0.06
(boron/vehicle
ratio)
• 15.7 ± 0.6 μg B/ml
• 0.15
(boron/vehicle
ratio)
boron concentrations were determined by ICP-MS
Micelle Formation
Bpin-PLA-PEOz
PBAD (Phenyl
boronic acid
derivative)
The boron bearing diblock
copolymers, once form the
micelles, could load
additional boron
compounds, PBAD in this
research.
Cell Viability by MTT assay
Cell viability after 48 h of incubation with the Bpin, PBAD, Bpin-PLA-PEOz
and Bpin-PLA-PEOz/PBAD; error bars are mean ± SD (n=6). *Significantly
different between PBAD and Bpin-PLA-PEOz/PBAD at the indicated
concentration (p < 0.05).
200
Hydrodynamic diameter
(nm)
Hydrodynamic diameter
(nm)
Stability of Micelle in Storage Condition
150
100
50
0
0
20
40
Time (day)
60
200
150
100
50
0
0
20
40
Time (day)
Size alteration of PLA-PEOz (left) and Bpin-PLA-PEOz micelles (right)
The encapsulated micelles were deposited in 4 °C for 10days, 30days
and 50 days. Then the micelles were analyzed by DLS to investigate
the alteration of the particle size.
60
Nanomaterial strategies from the point-of-view of the cell
Hypoxia
a state of decreased O2 availability below critical thresholds, thus
restricting function of organs, tissue, or cells.
Tissue hypoxia results from:
1. O2 tension
low O2 partial pressure~ pulmonary diseases, high altitude
2. anemic hypoxia
reduced ability of blood to carry O2 ~ anemia, CO poisoning
3. circulatory hypoxia
reduced tissue perfusion ~ generalized or local
4. diffusional hypoxia
increase diffusion distance
5. cytotoxic hypoxia
intoxication, cyanide poisoning
Tumor Hypoxia
• Solid tumor
~ highly heterogeneous
~ exhibit oxygen tension, low pH, low glucose concentration
Kizaka-Kondoh et al. Cancer Sci. December, 2003
Oxygen Tensions in Various Tissue, Tumor
O2 pressure (mmHg)
Normal tissues
Murine brain
60
Murine muscle
42
Bone marrow
40-50
Normal liver
55
Normal breast tissue
65
Normal cervix
48
Head and neck tissue
43
Normal tissue
~ 50-80 mmHg
Tumors
Breast carcinoma
Solid tumors
28
<2.5
Murine Fsall fibrosarcoma
<5
Cancer of cervix
<12
Head and neck cancers
<10
Soft-tissue sarcomas
<10
Tumor
< 10-30 mmHg
Nitroimdazoles derivatives
Hypoxia-Specific Tumor Imaging with 18F-Fluoroazomycin Arabinoside
A
C
B
O2N
N
O
18
N
F
HO
OH
FAZAB
J. Nucl. Med. 2005
NO2
N
HO
O
N
OH
N
O2N
HO
F
N
N
O
N
NO2
NO2
HO
HO
O
N
N
NO2
N
N
NO2
O2N
O
I
N
N
NO2
O
N
OH
I
Br
N
I
N
I
N
F
Euro. J. Nucl. Med. 1995
Bioreductive Metabolism of Nitroimidazole
Hydrophilic
Can Not penetrate membrane
OUT
M
E
M
B
R
A
N
E
Stock in cell
IN
.
O2-
entry
R- NO2
R- NO2
efflux
futile
cycle
1e
reduction
oxidative
.
R- NO2-
damage
.
1e
R-NO
R-NO2H
protonation
disproportionation
H+
hypoxia
decomposition
.
R
(ox)DNA+T-
DNA(-T)
O2
NO2
..
R-NO2H
H+
.
R
2e
H2 O
R-NO2
R-NHOH
2e
R-NH2
Hypoxia Targeting Nanodelivery
Size 
affects the biodistribution profile and therapeutical
bechaviour of the system
Nano-size 
penetration, cellular uptake, targeting
Normal tissue
•active
•passive
Hypoxia Tumour
tissue
Intratumoural
delivery of NPs
Defective
lymphatic
drainage
Lymph node
Dis-organized and
leaky tumour
endotelium
Normal vessels
with tight
endothelium
EPR-effect
Enhanced permeation
and retention effect
Synthetic Scheme of Bpin-PLA-PEOz-NIm
Strategy of Copolymers Synthesis
Synthesis of (Bu4N)[B12H11O(CH2)2O(CH2)2OH]
Polymers
Mw
Mn
PDI
B-PLA-OH
3175
2231
1.42
Nanodiamonds
• Chemically inert
– however, can be surface-functionalized
• Biocompatible
– low cytotoxicity
• Detectable – green fluorescence
– by confocal and flow cytometry
Applications of Diamond Materials
1. Tribology
2. Drug Delivery
3. Bioimaging
4. Tissue Engineering
5. Nanocomposites for Filler Materials
Mochalin, V. N. et al. Nat. Nanotechnol., 2011, 209, 11-23
Functionalized Nanodiamonds
Base on Organic Chemistry ~
Create New Nanodiamonds platform for Bioconjugate
Functionalized Nanodiamonds
Base on Organic Chemistry ~
Create New Nanodiamonds platform for Bioconjugate
S
O
HO
H2N
OH
O
BH3, THF
O
NH2
SH
HO
OH
1. HBr, HOAc, D
HS
SH
HO
OH
2. NaOH
3. H2SO4
HS
SH
O
O
O
OH
COOH
SH
N%
C%
H%
S%
ND
3.411
84.729
1.011
0.287
ND-COOH
3.316
82.911
1.047
0.373
ND-OH
1.721
78.254
1.080
0.023
ND-SH
2.970
79.844
0.923
3.305
Propose mechanism
NH2+
S
OH
H+
-H2O
+
H2N
S
NH2
NH2
OH H2O
SH
S
H+
- CO3- NH3
Accepted by ACS Applied Materials & Interfaces
XPS of Thiolated Nanodiamonds and Carbon Nanotube
Thiolated Nanodiamonds
C1s
-Csp3-SO2X
O1s
-Csp3-SH
OKLL
S2sS2p
1200
1000
800
600
400
200
0
B.E.(eV)
180
175
170
165
160
155
B.E.(eV)
Thiolated Carbon Nanotube
C1s
Survey CNT-SH
O1s
S2s S2p
C1s
Survey CNT-OH
O1s
S2s S2p
1200
1000
800
600
B.E. (eV)
400
200
0
Accepted by ACS Applied Materials & Interfaces
TEM of Thiolated Nanodiamonds (100 nm) with Au-NP
Accepted by ACS Applied Materials & Interfaces
Boron-containing Nanodiamonds
Boron Cage-containing Nanodiamonds
18th International Congress on Neutron Capture Therapy in Taiwan in 2018
Boron Neutron Capture Therapy – Best New Concept Therapy
October 28th – November 2nd, 2018 | Taipei, Taiwan
Thank you very much for your attention.
Taiwan is looking forward to seeing you
in 2018!