1. 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

2. Outline Nanomedicine Boron Containing Polmeric nanoparticle for BNCT
PLA-PEOz-B-pin
PLA-PEOz-B-cage
Boron Containing Nanodiamonds for BNCT
Boron Nanoparticles for BNCT

3. Nanomedicine: Nanotechnology to Medicine
Apply Nanotechnology:
Maintain drug activity.
Increase water solubility.
Delivery drug by nanoparticle
Increase drug efficiency
Traceable or directable drug
Nanomedicine
Use of nano-materials in medicine  unique medical effects

4. 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

5. The enhanced permeation and retention (EPR) effect
Nature Reviews Cancer 12, (January 2012)

6. 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

7. Development of BNCT Drug Deliver System Based on Polymeric Material PLA-PEOz

8. Polylactide, PLA Polyoxazoline
Biocompatible
Biodegradable
Low immunogenicity
Good mechanical properties
FDA-approved for clinical use
Hydrophobic
Pseudopolypeptides
Adjustable
Biocompatible
Low immunogenicity
Stealth
FDA-approved as a food contact agent
Hydrophilic

9. Strategy of Copolymers Synthesis

10. Synthetic Scheme of Bpin-PLA-PEOz

11. Characterizations of synthesized polymers
Table1 Characterizations of synthesized polymers
polymer
Mna
Polydispersity index (PDI)b
Yield (%)
CMC (wt %)
Bpin-PLA
7218
1.21 98
Bpin-PLA-PEOz
14247
1.24 62
7×10-4
a Estimated by 1H NMR . b Estimated by GPC.

12. Boron Content of Bpin-PLA-PEOz micelle
Synthesis of
Phenylboronic acid derivative
(PBAD)
Bpin-PLA-PEOz
6.06 ± 0.3 μg B/ml
0.06 (boron/vehicle ratio)
Bpin-PLA-PEOz/PBAD
15.7 ± 0.6 μg B/ml
0.15 (boron/vehicle ratio)
boron concentrations were determined by ICP-MS

13. 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.

14. 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).

15. Stability of Micelle in Storage Condition
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.

16. Nanomaterial strategies from the point-of-view of the cell

17. 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

18. Tumor Hypoxia Solid tumor ~ highly heterogeneous
~ exhibit oxygen tension, low pH, low glucose concentration
Kizaka-Kondoh et al. Cancer Sci. December, 2003

19. 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
Tumors
Breast carcinoma 28
Solid tumors
<2.5
Murine Fsall fibrosarcoma
<5
Cancer of cervix
<12
Head and neck cancers
<10
Soft-tissue sarcomas
Normal tissue
~ mmHg
Tumor
< mmHg

20. Nitroimdazoles derivatives
Hypoxia-Specific Tumor Imaging with 18F-Fluoroazomycin Arabinoside
J. Nucl. Med. 2005
Euro. J. Nucl. Med. 1995

21. Bioreductive Metabolism of Nitroimidazole
Hydrophilic
Can Not penetrate membrane
Stock in cell
hypoxia

22. 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
Dis-organized and leaky tumour endotelium
EPR-effect
Enhanced permeation and retention effect
Normal vessels with tight endothelium
Lymph node

23. Synthetic Scheme of Bpin-PLA-PEOz-NIm

24. Strategy of Copolymers Synthesis

25. Synthesis of (Bu4N)[B12H11O(CH2)2O(CH2)2OH]
Polymers Mw Mn
PDI
B-PLA-OH
3175
2231
1.42

28. Nanodiamonds Chemically inert Biocompatible
however, can be surface-functionalized
Biocompatible
low cytotoxicity
Detectable – green fluorescence
by confocal and flow cytometry

29. 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

30. Functionalized Nanodiamonds
Base on Organic Chemistry ~
Create New Nanodiamonds platform for Bioconjugate

31. Functionalized Nanodiamonds
Base on Organic Chemistry ~
Create New Nanodiamonds platform for Bioconjugate
1. HBr, HOAc, D
2. NaOH
3. H2SO4
BH3, THF 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
Accepted by ACS Applied Materials & Interfaces

32. XPS of Thiolated Nanodiamonds and Carbon Nanotube
Thiolated Carbon Nanotube
Accepted by ACS Applied Materials & Interfaces

33. TEM of Thiolated Nanodiamonds (100 nm) with Au-NP
Accepted by ACS Applied Materials & Interfaces

34. Boron-containing Nanodiamonds

35. Boron Cage-containing Nanodiamonds

36. Thank you very much for your attention.
18th International Congress on Neutron Capture Therapy in Taiwan in 2018
Boron Neutron Capture Therapy – Best New Concept Therapy
October 28th – November 2nd, | Taipei, Taiwan
Thank you very much for your attention.
Taiwan is looking forward to seeing you in 2018!