Irradiation Characteristics of D2O Facility in KUR with Low-enriched Uranium Fuel (II)CO7-1
Y. Sakurai and H. Tanaka
Research Reactor Institute, Kyoto University

Introduction: In Heavy Water Neutron Irradiation Facility (D2O Facility) of KUR, the neutron beams with thevarious energy spectra are available [1]. This facility has
been utilized mainly to boron neutron capture therapy(BNCT). In February 2006 just after the suspension of
the KUR operation started, the total number of BNCT
clinical irradiation reached to 275. In May 2010, BNCT
clinical irradiation restarted concurrently with the KUR
operation restarted. After the restart, 102 BNCT irradiations have already been carried out as of May 2012.
As this facility is a larger facility, it is expected that theirradiation characteristics are influenced by the conditions of the KUR fuel. Then, we performed there-evaluation of the irradiation characteristics.

Fig. 1. Neutron energy spectra after the fuellow-enrichment for CO-0000-F, OO-0000-F andOO-0011-F modes.

Methods: The neutron energy spectra were evaluatedfor the three standard irradiation modes of epi-thermal(CO-0000-F),
mix(OO-0000-F)
andthermal
(OO-0011-F). The evaluation was performed by multi
activation foil method using gold, indium, aluminum
nickel, etc.. The unfolding was done using an unfolding
code, MAXED [2]. The nominal spectra before the fuellow-enrichment were used as the initial guesses.
Results and Discussions: Figure 1 shows the evaluation results of the neutron energy spectra at the standard
irradiation position for the three irradiation modes. The
averaged data obtained from three irradiation experiments for the respective modes are drawn. The uncertainties for the three irradiations were within plus/minus 20%
in the thermal neutron range, 15% for the epi-thermal
neutron range and 20% for the fast neutron range.
Figure 2 shows the comparison for neutron energy spectra in CO-0000-F mode between before and after the fuellow-enrichment. It is found that the spectrum after the
low-enrichment is a little softer. It is found that the neutron beam intensity after the low-enrichment is overalllarger. It was confirmed that the beam intensity increased
at 10 to 20% for the fast neutron range, 20 to 30% for the
epi-thermal neutron range.
Conclusion: It was confirmed that the neutron energy
spectra became a little softer and the neutron beam intensity increased 10 to 30% after the fuel low-enrichment.
However, it was concluded that these changes in the irradiation characteristics due to the fuel low-enrichment
were not so larger for the influence on the BNCT clinicalirradiation, from the viewpoint of the relative values.

Fig. 2. Comparison for neutron energy spectra in
CO-0000-F mode between before and after the fuellow-enrichment.

REFERENCES:[1] Y. Sakurai et al., Nucl. Instr. Meth. A 453 (2000)
569-596.
[2] M. Reginatto et al., Nucl. Instr. Meth. A 476 (2002)
242-246.

採択課題番号 23006
KUR 燃料低濃縮化後の重水中性子照射設備の特性評価（京大・原子炉）櫻井良憲、田中浩基、齊藤 毅、吉永尚生（京大院・工）藤井孝明、上石達也、内田良平

共同通常

CO7-2

Short-Term Outcome of BNCT for Human Clear Cell Sarcoma in Mouse Model

T. Fujimoto, T. Andoh1, Y. Sakurai2, M. Suzuki3,
H. Ichikawa11 and K. Ono2
Hyogo Cancer Center,
Department of Orthopaedic Surgery
1
Faculty of Pharmaceutical Sciences,
Kobe Gakuin University
2
Department of Radiation Life Science,
Research Reactor Institute, Kyoto University
3
Particle Radiation Oncology Research Center,Research Reactor Institute, Kyoto University
INTRODUCTION: Clear cell sarcoma (CCS) of tendons and aponeuroses is a rare and malignant tumor ofpoor prognosis, with a predilection for young adults [1, 2].
The standard treatment is still wide surgical resection
because chemotherapy and radiotherapy are not effective.
Clearly therefore, new therapeutic strategies are required.
Recently, boron neutron capture therapy (BNCT) with theuse of p-boronophenylalanine (BPA) for malignant
melanoma has provided good results [3]. Since BPA is
associated with the production of melanin, boron is
ultimately accumulated in tumor cells. CCS also produces melanin, and with the use of a cultured human CCS
cell line and through human CCS-bearing animal studies
we have shown that high accumulation of 10B is potentially favorable in BNCT [4, 5]. Consequently, in thisstudy, we evaluated the efficacy of BNCT for CCS withthe use of human CCS cell line-bearing nude mice andpropose a new therapeutic option for the treatment of
CCS.
EXPERIMENTS: (1) Tumor cell line: Cells of human
CCS cell line MP-CCS-SY [6] were grown in RPMI
1640 and DMEM with fetal bovine serum in a 5% CO2
humidified incubator at 37˚C.
(2) BNCT for CCS (MP-CCS-SY)-bearing animals: Allanimal experiments were carried out according to theregulations of the Animal Care and Use Committee.
MP-CCS-SY cells were subcutaneously transplanted into
the left femoral region of BALB/c nude mice. Fourweeks thereafter, the animals were divided into fourgroups of 4 each. Of nonirradiated groups A and B, saline
was administrated to group A and BPA-Fr (500mg
BPA/kg) was administrated to group B through the femoral vein, under anesthesia, on day 0. Similarly saline(group C) and BPA-Fr (500mg BPA/kg) (group D) wereadministrated to irradiated groups C and D, respectively,
and then the two groups were immediately placed in achamber for thermal neutron irradiation. Thermal neutrons (1MW) were delivered from the dorsum of themouse, in the heavy water facility at KURRI. LiF tiles
were used to shield parts of the body other than the left
leg. The size of the tumor was measured in groups A andB from day 0 to day 28, and in groups C and D from day

0 to day 23. Thin sections from formalin-fixed and paraffin-embedded samples of the tumor mass resected on thelast day of measurement were stained with hematoxylin-eosin (HE) for histological evaluation.
RESULTS: The size of the tumor decreased
time-dependently in only group D. Finally, on day 23, thetumors in 2 mice of group D had disappeared, and thetumors in the two remaining animals were excised. Although the size of the tumor increased in groups A, B andC, the growth was slightly suppressed in group C compared with that in the other two groups. There was nodifference between groups A and B. Histological examination of the sections of group D revealed that the tumor
mass had disappeared completely 2 tumors and hadchanged into granulation tissue 2 tumors — only the tumor mass was selectively destroyed with no damage tonormal surrounding tissue [Fig.1]. On the other hand,saline-administrated irradiated group C [Fig.1] showedno tumor death and no normal tissue damage.

[Fig.1] Left: CCS tumor mass resected from group C (hotcontrol) on day 23. Tumor cells are observed in the subcutaneous area. No tumor cells or skin were damaged.
Magnification (80×). Right: CCS tumor mass resectedfrom group D (BNCT) on day 28. No tumor cells areobserved. The connective tissue is replaced by granulation. There is no damage to normal tissue surrounding thetumor. Magnification (80×).
REFERENCES:[1] F.M. Enzinger, Cancer, 18(1965) 1163-1174.
[2] W. Deenik et al., Cancer, 86(1999) 969-975.
[3] Y. Mishima et al., Lancet, 12(1989) 388-389.
[4] T.Fujimoto et al., Appl. Radiat. Isot.
69(2011)1713-1716.
[5] T.Andoh et al., Appl. Radiat. Isot.
69(2011)1721-1724.
[6] H. Moritake et al., Cancer Genet. Cytogenet.,
135(2002) 48-56

採択課題番号 23009
明細胞肉腫に対するホウ素中性子捕捉療法の検討
共同通常（京大・原子炉）鈴木 実、櫻井良憲、小野公二（神戸学院大・薬）安藤 徹、市川秀喜（兵庫県立がんセンター・整形外科）藤本卓也

CO7-3

Enhanced the Effect of Boron Neutron Capture Therapy: Design of Boron-containing
Nanoparticles with Highly Tumor-accumulating Character

Y. Nagasaki1-3, T. Yaguchi1, M. Oishi1, S. Sumitani1, H.
Murotani1, Y. Horiguchi2, M. Suzuki4, K. Ono4, H.
Yanagie5
1

Institute of Materials Science, Graduate School of Pureand Applied Sciences, University of Tsukuba
2
Master’s School of Medical Sciences, Graduate Schoolof Comprehensive Human Sciences, University of Tsukuba
3
Satellite Laboratory, International Center for Materials
Nanoarchitectonics (WPI-MANA), National Institute ofMaterials Science (NIMS)
4
Radiation Oncology Research Laboratory, Research
Reactor Institute, Kyoto University
5
Department of Nuclear Engineering and Management,
School of Engineering, University of Tokyo

Introduction:

The success of BNCT is dependent on the delivery systems to accumulate a sufficient quantity of 10B to tumor
tissues. Herein, we designed and prepared core polymerized and boron-conjugated micelles (PM micelles) composed of poly(ethylene glycol)-block-poly(lactide) copolymer bearing an acetal group at PEG end and amethacryloyl group at PLA end (acetal-PEG-b-PLA-MA)
and polymerizable boron cluster (1-(4-vinylbenzyl-closocarborane): VB-carborane. This micelle enables to prolong the circulation time and deliver the boron to the tumor tissues without leakage of boron clusters in the bloodstream because of the existence of covalent bonds between boron clusters and the PLA core of the micelles.

sodium (40 mg/kg) and placed in an acrylic mouse holder.
The mice were irradiated thermal neutrons for 37 min at a
rate of 1.6-1.8 x 1012 neutrons/cm2. Figure 1 shows theneutron irradiation to tumor bearing mice at Kyoto University Research Reactor. The BNCT effects were evaluated in terms of the tumor size.

Results and discussion:

The average diameter of the PM micelles (67.3 nm,
μ2/Γ2 = 0.113) was almost similar to that of the NPM micelles (60.2 nm, μ2/Γ2 = 0.119), suggesting that the corepolymerization process does not influence to the average
diameter and size distribution of the micelles. The loading content of boron atoms in the NPM and PM micelles
was determined to be 8.5 wt% (loading efficiency:
23.5 %) and 7.7 wt% (loading efficiency: 21.5 %), respectively. Note that the significant suppression of
growth of tumor volume in mice treated the 10B-enriched
PM micelles was observed from 12 days compared to themice treated normal saline (p < 0.01). On the other hand,
the mice treated the 10B-enriched NPM micelles and free
BSH showed tumor growth similar to the control. These
results indicate that the concentration of 10B atoms intumor tissues of the mice treated the 10B-enriched NPM
micelles or free BSH was insufficient for BNCT.[1]Experimental Methods:

The PM micelle was prepared from acetal-PEG-b-PLA-MA (PEG; Mn=5,600, PLA; Mn=5,100)
micelle encapsulating VB-carborane and radical initiator(azobisisobutyronitrile: AIBN) by solvent evaporation
method. The polymerization of the core was carried out
by thermo-induced radical copolymerization at 60 °C for
24 h. For comparison, NPM micelles encapsulating
VB-carborane were prepared by the same procedure asthat for PM micelles, without the addition of AIBN andheating. Neutron irradiation was carried out at Kyoto
University Reactor of Kyoto University Research Reactor
Institute. 10B-enriched PM and NPM micelles were prepared from 10B-enriched VB-carborane and acetal-PEG-b-PLA-MA by same procedure above.
10
B-enriched PM and NPM micelles solution injected intocolon-26 tumor bearing mice (n = 5) via the tail vein at adose of 15.6 mg 10B/kg 24 h before irradiation. As comparison, 10B-enriched BSH solution was injected 1 h before irradiation via the tail vein at a dose of 30.0 mg
10B/kg. The mice were anesthetized with pentobarbital

採択課題番号 23015

Fig. 1. Neutron irradiation to tumor bearing mice at Kyoto University Research Reactor

REFERENCES:[1] S. Sumitani et al., Biomaterials, 33 (2012) 3568

高い腫瘍選択性を有するボロン含有ナノ粒子の創製と共同通常
中性子捕捉療法への展開（筑波大・数理）長崎幸夫、矢口達也、大石 基、角谷省吾、室谷憲紀、堀口諭吉、尾崎佑樹、西澤信子、吉冨 徹、（京大・原子炉）鈴木実、小野公二（東大院・工）柳衛宏宣

CO7-4Development of Novel Boron Compounds for BNCT

2-

CO2H

H2N

10

B

HO

10

SH

2Na+

B

BSH

BPA

0.1

0.01

1a
1b
1c
BPA
CONT.

10

B

S

CO2H
n

+

2Na

0.1

0.01
1a
1b
1c
BPA
CONT
0.001

0.001
0

1
2
3
4
5
12
-2
Thermal Neutron Fluence (10 cm )

0

1
2
3
4
12
-2
Thermal Neutron Fluence (10 cm )C: SAS
1

0.1

0.01

1a
1b
1c
BPA
CONT.

0.001
1
2
3
4
12
-2
Thermal Neutron Fluence (10 cm )Fig. 2. Tumor cell killing effects of boron amino acids.
A: against C6 cell. B: against B16 cell. C: against SAS
cell.

NH2

2-

Fig. 1.

1

0OH

B: B16

A: C6
1

Surviving Fraction

INTRODUCTION: Although many kinds of boron
compounds such as amino acid, nucleic acid and liposome have been reported as boron boron carrier for boron
neutron capture therapy (BNCT) [1], only two compounds, p-borono-L-phenylalanine (BPA) and mercapto-closo-undecahydrododecaborate (BSH) of them,
are clinically used in cure of cancer with BNCT.
To develop practical materials utilizing 10B carrier, we
had already designed and synthesized new boron containing amino acids [2] such as C4-BSH-AA (n=2) (1a),C5-BSH-AA (n=3) (1b), C8-BSH-AA (n=6) (1c), which
includetheundecahydro-closo-dodecaboranylthio([10B12H11S]2--) unit by boron-sulfur-carbon bond connection in the side chain of α-amino acid (Fig. 1). Here, wereport the tumor cell killing effects of BSH-amino acids
1a-c.
Material and Method: Cultures were inoculated with
1.0 x 106 cells/dish, and cells were grown for 24 h in
DMEM. The medium was replaced with the each medium containing each boron amino acid (final concentration was 2.0 mM in each case). The cells were cultured

Surviving fraction

Graduate School of Life and Environmental Sciences,
Osaka Prefecture University
1
Research Organization for University-Community
Collaborations, Osaka Prefecture University
2
Research Reactor Institute, Kyoto University

averaging two gold foils symmetrically attached to thesurface of the column-shape tube along the direction ofincidence of thermal neutrons. After thermal neutron
exposure, 600 cells were placed in three Corning 60 mm
tissue culture dishes containing 3 mL DMEM to examine
colony formation. Seven days later, the colonies were
fixed with ethanol and stained with 0.1% crystal violet
for quantitative visualization by the naked eye.
To confirm the usefulness of BSH-amino acids 1a-c for
BNCT, we examined the tumor cell killing effects of BPA
and the compounds 1a-c against three kinds of tumor cellsuch as C6 (rat glioma), B16 (mouse melanoma), andSAS (human oral squamous cell carcinoma).

Surviving Fraction

M. Kirihata, Y. Hattori1, M. Mukumoto, S. Ueda, H. Murakami, M. Suzuki2, S. Masunaga2, K. Ono2

:

10

BH

1a - c (1a: n = 2, 1b: n = 3, 1c: n = 6)
Boron compounds

for 24 h, and the medium was removed by aspiration.
The cells were washed with PBS, harvested by trypsinization, and then cell numbers were counted. After centrifugation, the tripsin was removed by aspiration, and to
the residual cells was added DMEM. The suspension of
the cell in DMEM (5.0 x 103 cells/mL, 1mL) was irradiated with thermal neutron for 0 - 90 min in column-shape
tube. The thermal neutron fluence was determined by

RESULTS: As shown in Fig. 2, 1a-c showed higher
killing effects than that of BPA in the case of each tumor
cells. These results suggest that BSH-amino acids 1a-c
are useful for 10B carrier on BNCT.
REFERENCES:[1] Soloway, A. H.; Tjarks, W.; Barnum, B. A.; Rong, F.
G.; Barth, R. F.; Codogni, I. M.; Wilson, J. G. Chem. Rev.,
1998, 98, 1515–1562.
[2] Kusaka, S.; Hattori, Y.; Uehara, K.; Asano, T.; Tanimori, S.; Kirihata, M. Appl. Radiat. Isot., 2011, 69,
1768-1770.

採択課題番号 23017
ホウ素中性子捕捉療法の新規ホウ素薬剤の開発研究
共同通常（阪府大院・生命環境）切畑 光統、椋本 麻里、上田 さおり、上田 晋平、村上 広和 （京大・原子炉）小野 公二、増永 慎一郎、鈴木 実 （阪府大・地域連携）服部 能英Fluorescence-labeled closo-dodecaborate Lipid: Synthesis and its Liposome Formation for in 
CO7-5
vivo Imaging Targeting to Tumor on BNCT
H. Nakamura, N. Ueda, H. S. Ban, M. Ueno, S.
Tachikawa, M. Suzuki1, S. Matsunaga1 and K. Ono1
Faculty of Science, Gakushuin University
1
Research Reactor Institute, Kyoto University
INTRODUCTION: Recently much attention has been
focused on the liposomal drug delivery system (DDS)
using drugs-embedded nano carriers including liposomes,
micelles,proteinaceousorpolymer-conjugatedmacromolecules, and lipid particles.[1] This nanocarrier-based DDS is based on the enhanced permiability
and retention (EPR) effect discovered by Maeda andMatsumura. Abnormal architectures and impaired
functional regulation of newly formed tumoral bloodvessels lead to abnormal molecular and fluid transport
dynamics especially for macromolecular species. Thus
nano particles extravasate through these tumoral bloodvassels and accumulate in tumor.
The cytotoxic effect of boron neutron capture therapy
BNCT is due to the nuclear reaction of two essentiallynontoxic species, boron-10 and thermal neutrons. It has
been considered that the nano carrier-based DDS is an
attractive approach for efficient boron delivery system(BDS) to tumor on BNCT. We were the first to synthesize
boron cluster lipid. Although the first generation boron
cluster lipid showed acute toxicity against tumor bearing
mice,[2] the second generation boron cluster liposomes,
such as DSBL that possess the B12H11S moiety as thehydrophilic function, did not display the acute toxicity up
to 30 mg boron per kg body in mice.[3] In BDS, it is very
important to know detailed boron biodistribution intissues because the distrubution of tumor cells are often
heterogeneous and liposomes may spread in a certain
distance from tumoral blood vessels. In this paper, we
reportdevelopemntoffluorescent-labeled
closo-dodecaborane lipid (FL-SBL) and its liposome
formation.
EXPERIMENTS: PEGylated DSPC-liposomes wereprepared from DSPC, CH, and DSPE-PEG-OMe(1:1:0.11, molar ratio) by the reverse-phase evaporation(REV) method. Briefly, a mixture of DSPC, CH, and
DSPE-PEG-OMe were dissolved in 10 ml of chloroform/diisopropylether mixture (1:1, v/v) in around-bottom flask. To the lipid solution was added 5 mLof water, forming an emulsion. The emulsion wassonicated for 1 min, and then the organic solvent wasremoved under reduced pressure in a rotary evaporator at
37 °C, generating a suspension of liposomes. The liposomes obtained were subjected to extrusion using an extruder device. Purification was accomplished by ultra-

採択課題番号 23018

centrifuging at 200,000 g for 20 min. FL-SBL was dissolved in normal saline solution and the FL-SBL solution
were mixed with DSPC liposome solution. The mixture
was maintained at 25°C for 5 min and free FL-SBL wasremoved by ultracentrifugation at 200,000 g. The obtained FL-SBL-labeled liposomes were resuspended in
PBS. Liposome size and Zeta potential were measured
with an electrophoretic light scattering spectrophotometer(Nano-ZS, Sysmex, Japan). The compositions of FL-SBL
and DSPC in liposomes were calculated from data obtained by the simultaneous measurement of boron andphosphorus concentrations by inductively coupled plasma
atomic emission spectroscopy (ICP-AES, HORIBA, Japan). The fluorescence intensity of each sample wasmeasured at a range of emission-wave length from 475 to
650 nm using an excitation wavelength of 460 nm.
RESULTS: The results are summarized in Table 1. The
PEGylated DSPC liposomes displayed 97.22 nm in diameter with -29.3 mV of Zeta potential. With increase ofvolumes of the additive FL-SBL solution, the boron concentration in liposomes increased. The boron/phosphorus
concentration ratio increased roughly in proportion from
1.9 to 7.1% w/w without affecting their particle size.
However, addition of the FL-SBL solution increased thezeta potential of each liposome from -29.3 to -20.0 mV.
Table 1. Boron and phosphorus concentrations, particle size,
and zeta potential of the FL-SBL-labeled liposomes
Volumeof
FL-SBL
(L)
0
10
20
30
40

Boron Phosphorus
conc. conc. (ppm)
(ppm)

B/P
ratio
(%)

Particle
size
(nm)

Zeta
potential
(mV)

41.3
73.8
112.6
146.7

0
1.9
3.4
5.4
7.1

97.2±0.15
97.6±0.47
97.4±0.20
96.2±1.38
97.8±1.15－4.10±0.35－23.2±2.35－23.3±3.04－21.1±1.70－20.0±3.52

1876.22153.42169.22066.12054.5Preliminary in vivo imaging study of the boron
liposomes using colon 26 tumor-bearing mice showedthat the boron liposomes are delivered to the tumor
tissue but not distributed to hypoxic tissue which islocated far from blood supply in tumor. [4]
REFERENCES:[1] D. F. Emerich et al., Biomol. Eng, 23, 2006, 171.
[2] (a) H. Nakamura et al., Chem. Commun. 2004, 1910.
(b) Y. Miyajima et al., Bioconjugate Chem. 17, 2006,1314.
[3] (a) J.-D. Lee et al., Org. Lett. 9, 2007, 323. (b) Ueno,
M. et al., Bioorg. Med.Chem. 18, 2010, 3059.
[4] H. Nakamura et al., Org. Biomol. Chem. 10, 2012,1374.

中性子捕捉治療用新規ホウ素薬剤の開発（学習院大・理）中村 浩之、上田記子、潘 鉉承、上野 学、立川 将士（京大・原子炉）鈴木 実、増永 慎一郎、小野 公二

共同通常

CO7-6

Basic Investigation of BNCT with Novel Boron Compound for 0alignant 3leural
Mesothelioma Cells

C.M. Lee1, 2, N. Sougawa1, M. Sasai3, M. Suzuki4, S.
Masunaga4 and Y. Sakurai4
Graduate School of Medicine, Osaka University
1
Department of Surgery
2
Department of Health Economics and Industrial Policy
3Medical Center for Translational Research4
Research Reactor Institute, Kyoto University
INTRODUCTION: Malignant pleural mesothelioma(MPM) is an aggressive and refractory cancer of thepleural linings of the lungs and chest wall caused by asbestos exposure. The therapy of MPM is required themultidisciplinary techniques: surgery, chemotherapy , andradiotherapy. However, surgical therapy, especiallyextrapleural pneumonectomy is limited to the locally advanced MPM. The chemotherapeutic regimens have resulted in an improved tumor response, but they have notbeen breakthrough tools yet. Radiotherapy is limited utility because the extensiveness of the tumor requires large
fields and it is impossible to administer tumoricidal dose
without injuring the adjacent lung and mediastinal organs[1, 2].
Boron neutron capture therapy (BNCT) could be abreakthrough strategy to treat MPM, because it is suitable
for the treatment of diffuse and invasive tumor. However,the success of BNCT depends upon the selective deliveryof 10B-atoms to tumor cells to complement the attenuation of thermal neutron. BNCT is done against the tumorsthat occupied near the body surface area efficiently.
Concerning BNCT for deep site tumor (MPM), novelcompound that possesses high affinity to tumor cells thanconventional boron compounds are craved for BNCT for
deep site tumors [3]. We developed novel boron compound that bound with MPM cells preferentially, and
BNCT with this novel compound showed significant efficiency compared with other boron compound including
conventional boron compound. We examined in vitro
experiments: boron neutron capture reaction with thisboron compound at KUR.
EXPERIMENTS: The murine malignant pleural mesothelioma cells were seeded into 35 mm dish, and novel
boron compound(HA-BND-S) or BSH was administeredinto cells for 24 hrs or 1 hr. The concentration of boron
compounds was 0.003 ppm per cell. After discarding boron compounds and washing with PBS, each group ofcells was set into Eppendorf tubes, and neutron irradiation was performed at JRR4 for 23 min at a rate of 1.8 x
1012 neutrons/cm2. After neutron irradiation, the cells

採択課題番号 23025

were washed with PBS, and seeded into 96 well plates atthe concentration of 2,000 cells/200 L/well. The cytotoxicity of BNCT with born compounds for MPM cells
was measured with Cell Cycling-8 kit using tetrazolium
assay.
RESULTS: As shown in Fig. 1, HA-BNS-S had thehighest cytotoxicity compared with the other boron
compounds including no discard of BSH group, despite
discarding before neutron irradiation.

Fig. 1. Cytotoxicity of BNCT with each boron
compound for MPM cells. Neutron irradiation was
done at a rate of 1.8 x 1012 neutrons/cm2 at JRR-4.
The cytotoxicity was
REFERENCES:[1] D. C. Rice et al., Int J Radiat Oncol Biol Phys, 69,(2007) 350-357.
[2] A. M. Allen et al., Int J Radiat Oncol Biol Phys, 65,(2006) 640-645.
[3] H. Fujii et al., Radiat. Oncol., 20, (2011)

深部腫瘍性病変に対するホウ素中性子捕捉療法の共同通常
治療効果改善のための新規ホウソ製材の開発（阪大・医）李 千萬、寒川延子、笹井雅夫（京大・原子炉）鈴木 実、増永慎一郎、櫻井良憲

CO7-7 Pilot Application of Gadrinium-Platinum Nanomicelles to Gd- Neutron Capture Therapy
H. Yanagie, N. Dewi, N. Iyomoto, Y. Sakurai1,
M. Umano2, T. Uryuda2, T. Nagasaki2, M. Suzuki3,
Y. Sakurai3, S. Masunaga3and K. Ono3
Department of Nuclear Engineering & Management, The
University of Tokyo
1
Cooperative Unit of Medicine & Engineering, The University of Tokyo Hospital2
Department of Bioengineering, Osaka City University
3
Research Reactor Institute, Kyoto University
SUMMARY : Considering the benefits that Gd-NCT
offers, especially its possibility of improving patient’s
quality of life by reducing irradiation time, and also
the prospective combination of chemotherapy andradiotherapy to increase treatment effectivity. Our
purpose in this research is to evaluate the feasibility ofgadolinium-platinum nanomicelles as Gd-NCT agent
by performing in vivo experiment using Colon-26
bearing mice.
INTRODUCTION: The use of gadolinium as neutron
capture therapy (NCT) agent has been getting attention
because of its highest neutron cross section (255 000
barns) and the fact that secondary particle produced after
capturing neutron have total kinetic energy about 3 timesof that produced by 10B. On our previous experiment wehave shown that gadolinium-entrapped liposome is apotential agent for Gd-NCT. In this current study, ourobjective is to evaluate the possibility of using gadolinium-platinum nanomicelles developed by Kaida et. al.1)
as Gd-NCT agent considering that there is a promisingcombined effect between gadolinium as NCT agent andplatinum-based drugs as chemotherapy agent. It is expected that we could enhance tumor-cell killing effectand reduce negative side effect of the treatment, which
eventually give the possibility to improve patient’s quality of life.
EXPERIMENTS: In this experiment, we preparedcolon26-bearing mice for in vivo experiment and performed thermal neutron irradiation at Kyoto University
Research Reactor Institute with a collimated neutron
beam. Two hundred microliter of Gd-Pt nanomicelles was

採択課題番号 23034

injected into each mouse and qualitative analysis was
performed by using MRI facility in Kashiwa campus, The
University of Tokyo, while quantitative analysis was
performed by using ICP-MS at Juntendo University.
Thermal neutron irradiation was performed with 1 x
10E12 n/cm2 thermal neutron fluence and then antitumor
effect was evaluated on the basis of the change in tumour
growth and survival rate of the mice.
RESULTS: Gd-Pt nanomicelles-injected mice revealed
about three times higher tumor growth suppression compared to non-treated group as shown in Fig.1. Some results of tumor growth suppression revealed that toxicity
is existed after neutron irradiation in treated groups with
Gd-Pt nanomicelles. There is possibility of platinum
photoactivation caused by reaction between secondary
particles produced after Gd-NCR such as gamma rays
and x-ray with energy above K-edge energy of platinum.
Moreover, as the nanomicelles compound bind gadolinium and platinum together very close, the possibility of
secondary reaction to occur is higher since the effective
solid angle is very large. The effectivity of Gd-Pt
nanomicelles as Gd-NCT agent is quite promising andfurther investigation is necessary to determine optimum
combination between Gd-Pt concentration and neutron
fluence applied for the treatment.

Fig. 1. Comparison of tumor growth suppression between Gd-Pt nanomicelles-injected group andnon-treated (control) group
REFERENCES:[1] Kaida et al.: Cancer Res 70: 7031-7041 (2010).
[2] De Stasio et al: Cancer Res, 61, 4272-7, (2001).

中性子捕捉療法法の一般外科領域癌への展開に向けた
基礎的・臨床的研究

共同通常（東大・原子力国際専攻）柳衛宏宣、伊豫本直子、Novriana Dewi（東大・獣医）柳川将志、飯塚智也（東大・心臓外科）櫻井由里子、毛利きくえ（宏仁会メディカルシティ東部病院）東 秀史、
瀬口浩司、太田嘉一（大阪市大・バイオ）長崎 健、馬野正幸、瓜生田貴聡（京大・原子炉）小野公二、増永慎一郎、鈴木 実、櫻井良憲

CO7-8Boron Neutron Capture Therapy for Advanced Head and Neck Carcinoma

T. Aihara, M.D.1 J. Hiratsuka, M.D.2
N. Morita, M.D.1 M. Uno, M.D.1 A.
Maruhashi, Ph.D3 K. Ono, M.D.3DQGT. Harada,
M.D.1
1

Departments of Otolaryngology Head and Neck Surgery,and 2Radiation Oncology, Kawasaki Medical School,
Kurashiki, Japan
3
Radiation Oncology Research Laboratory, Research
Reactor Institute, Kyoto University, Osaka, Japan

ation at the KUR with a reactor power of 5 MW. The
irradiation field was large enough to cover the target area
for the neutron beam (10 cm×10 cm). 3) Neutron flux
measurement using gold wire 15 min. after the start ofirradiation. 4) Optimization of the neutron dose based on
the measured blood 10B concentration and neutron flux.
The tumor dose and normal tissue dose calculated ranged
from 20.0 to 30.0Gy-Eq and from Less than 15Gy-Eq,
respectively. The median duration of observation is 6.5
months after BNCT (4-10 months).

INTRODUCTION: Advanced head and neck carcinoma (AHNC) and recurrent head and neck cancer(RHNC) are often radio-/chemo-resistant and show extensive growth, requiring a wide resection including surrounding normal tissues. To avoid severe impairmentof head and neck structures, it is necessary to explore
new treatment for AHNC. Mishima first proposed employing boron neutron capture therapy (BNCT) for malignant melanomas utilizing the specific melanin synthesis activity of melanoma cells1). Kato et al.2) began
BNCT using both BSH (Na2B12H11SH) and BPA (para-boronophenylalanine) for recurrent parotid gland carcinoma for the first time and reported excellent preliminary results. On the basis of the encouraging results of
their pioneering clinical trial, our many years’ experience
with melanoma BNCT and the trend toward emphasizing
the quality of life after treatment, we also started treating
our patients with BNCT using BPA alone3)4). In thisreport, we summarize our clinical results of AHNC andRHNC cases treated by BNCT using BPA alone in 2011.

RESULTS: Six patients demonstrated regional partial
remission (PR), and two patients were no change (NC).
One patient out of NC cases was operated on after irradiation. Six patients are living with disease. Two patients were death of carcinoma. All patients had no acuteand/or chronic severe complications such as skin ulcers,
xerostomia, and palsy of the cervical spinal cord.

MATERIALS and METHODS: Between April 2011
and September 2011, Two patients with AHNC (Onemaxillary salivary duct carcinoma, one left jugular leiomyosarcoma), 5 patients with RHNC (3 recurrent SCC,
one parotid gland adenoidcystic carcinoma, one cervicalamelanotic melanoma), and one patient with skin melanoma received BNCT using BPA alone in the condition
that a tumor/normal tissue boron concentration ratio (T/Nratio) exceeded 2.5 based on 18F-BPA PET studies. Acceptable criteria is followings: (1) With head and neckT3/T4 tumors that surgical treatment is not indicated, (2)The depth of tumor, less than 6cm, and without distantmetastases, (3) PS≦ 2, (4)T/N ratio ≧2.5 using
18
F-BPA・PET, (5)Consent to perform BNCT, (6)With theapproval of our Medical Ethics Committee. The procedures for BNCT using BPA were as follows: 1) Intravenous administration of BPA-fructose complex(500mg/Kg.BW) for 2.5 to 3 hour and blood sampling at
the time of just finished BPA-drip and just before irradiation. The 10B concentration in the blood was measured byprompt γ-ray spectrometry. 2) Epithermal neutron irradi-

REFERENCES:[1] Y Mishima. Neutron capture treatment of malignant
melanoma using 10B-chlorpromazine. Pigment Cell Res.
1:215-221,1973[2] I Kato, et.al. Effectiveness of BNCT for recurrent
head and neck malignancies. Applied Radiation and Isotopes 61:1069-1073,2004[3] T.Aihara, et al. First clinical case of boron neutron
capture therapy for head and neck malignancies utilizing
18F-BPA PET. Head and Neck 28 p850-855; 2006
[4] T.Aihara, et al. BNCT for advanced or recurrent headand neck cancer. New Challenges in Neutron capture
therapy.：25-29 2010

CONCLUSION: Our results validate the efficacy of
BNCT in the treatment of patients with AHNC and
RHNC. Although this is a clinical report of only eightpatients, additional long-term follow-up should be required to assess this treatment. We have estimated T/Nboron ratio using 18F-BPA-PET in every cases. The T/Nratios measured are the values of BPA alone. If T/Nratio was more than 2.5, according to our adaptation, it isthought that therapy effect is good. We believe that headand neck tumors are suitable for BNCT and that suchexcellent results will have a great impact on patients inthe near future.

採択課題番号 23035
中性子捕捉療法適応癌腫の治療プロトコルの確率
共同通常（川崎医大）平塚純一、原田 保、粟飯原輝人、宇野雅子、牧野栄一、笹岡俊輔、森田倫正
小西圭、神谷伸彦（京大・原子炉）小野公二、丸橋 晃

CO7-9Carboranyl Glucose and Ribose Derivatives for BNCT

M. Takagaki, S. Masunaga1, I. Snajdr2, R. Satapathy2 andNS. Hosmane2
College of Nursing, Aino Univ
1
Research Reactor Institute, Kyoto University2
Dept Chem & Biochem, Northern Illinois Univ

Glucose and/or ribose attached carborane and theirisomers has been investigated for boronated agentsof BNCT［1］.
Although carborane attached 5-thio-d-glucose andcarborane attached 5-thio-d-glucopyranose werehighly toxic, toxicity could be decreased in 78% by
a replacement form of carborane attached5-thio-D-glucose with ribose, carboranyl ribose(fig.1).

Fig. 1.

Carboranyl ribose.

However IC50 was still high, 5.77 x 10-2 mM. And,furthermore, uptake of them into SCCVII tumor
cells was very low and No any in-vitro BNCT
effect were observed even even after oreincubationof B-loading in maximum tolerable B concentration.
(Fig. 2)
Recently, toxicity could be decreased to 1/100 by
modifying carboranyl-thio-D-glucose and cesium
salt form of carboranyl ribose. The uptake of theminto SCCVII tumor cell was observed. In-vitro
BNCT and bio distribution study has been underinvestigation at KUR (#2457).
Temporally Summary
1. A glucose conjugated carborane; carborane
attached
5-Thio-D-Glucopyranose,wasextremely toxic.
2. A form of ribose conjugated carborane
reduced its toxicity in 78%, but still highly
toxic.
3. The mechanism of toxicity is unknown, but it
might be caused by a carborane-related
chemical toxicity.
4. In-vitro BNCT has been under investigation ina new protocol using newly synthesized novel
models of glucose and/or ribose conjugatedcarborane.
［1］Khan, FA., Mangeswara, GH., Satapathy, R., K.
Parasuraman.: Serendipitously discovered diazomethanemediated novel molecular rearrangements of norbornyl
-ketohemiketals. Org Lett, Vol. 9(8), 1581-1584, 2007.

Fig.2. Survival fraction for glucose conjugated carborane.

採択課題番号 23036
悪性脳腫瘍のための熱外中性子捕捉療法の基礎的研究（藍野大学）高垣政雄（東邦大学・医）東丸貴信（京大炉）増永慎一郎

共同通常CO7-10

BNCT Case Report of Malignant Brain Tumor 2011

T. Tchizawa, M. Takagaki1, H. Tanaka2, Y. Sakurai2, M.
Suzuki2, A. Maruhashi2, N. Kondo2, S. Masunaga2 andK. Ono2
Department of Neurosurgery, Kanto Rosai Hospital
College of Nursing, Aino University
2
Research Reactor Institute, Kyoto University
1

Case Report: Grade-III glioma 56y female wastreated via BPA-based BNCT on Oct. 6, 2011.
The patient initially diagnosed as Grade III glioma
for her temporally dysarthria and numbness of her
upper extremity and face. Those symptoms
disappeared for 10 minutes. However the patient
had the same symptoms on the next morning andconsulted a doctor.
The patients showed no any neurological deficit
except dysarthria and NIHSS=1.
The magnetic resonance imaging showed airregular round shaped tumor in less than 3cm
diameter in the superficial convexity of the rightfrontal lobe (Fig. 1).

2 days after BNCT, the patient showed conscious
dist. (GCS:E2-3, V4, M6, somnolence) and left
hemiparesis (MMT UE 1/5, LE 2/5) for the rapid
enlargement of the tumor cystic cavity andperipheral edema shifting the mid-line structures
after BNCT. The patient emergently received cyst
drainage via Ommaya reservoir and condition
drastically improved.
The patient received TMZ, 120mg/day 12 days
after BNCT, and also received a booster radiation
10Gy (Cyber Knife) on the deepest lesion which
dose was less than 30Gy. Then the cytologicalfindings of the repeatedly removed fluid of thecystic cavity improved Class IV to Class II.
The current KPS was stable 70 at home. The cystic
cavity was still enhanced in stable configuration 7
Mo after BNCT (Fig. 2).

Fig. 2. Magnetic resonance Gd-DTPA enhanced image
7Mo after BNCT. Tumor remained with ring-likeenhancement lesion, although the cyto-hystologicalserial findings the cystic fluid changed into benign.

Fig.1. Magnetic resonance image of the patient’s tumor before
BNCT. The tumor located superficially in the parietal
convexity with peripheral low density rim.

The spatial radiation measurement of thepost BNCT head of this patient: Post BNCT
spatial γ-ray was serially measured by Aloka
NaI(Tl) scintillation survey meter. The
background γ-ray dose in the word was
0.07-0.08 µSv/h. The figures shows the treatment
and/or nursing close to the patient’s head should bemanaged within minimum duration for 2 days after
BNCT.

The open biopsy proved Grade-III glioma initiallybut the definite diagnosis was changed to be
Grade-IV glioma via precise inspection of thebiopsy specimen.
1 month after the onset, the patient gradually
showed aggravation of the numbness and then thepatient was treated by BNCT under the maximum
tolerance normal brain dose, T/N=2.5, av. 20ppm
10
B.

Table 3 Serial measurement of the special γ-ray
radiation dose of the post BNCT head.

採択課題番号 23037	
 	
 	
  	
 悪性脳腫瘍のための熱外中性子捕捉療法の臨床的研究	
 	
  	
  	
  	
  共同通常	 （藍野学院）高垣政雄（関東労災病院）立澤孝幸（シミズ病院）大山憲治	
 （京大・原子炉）小野公二	
 CO7-11

Hyaluronan-Conjugated Liposomes as Carrier for Oral Squamous Cell Carcinoma
in Boron Neutron Capture Therapy

Y. Ito, Y. Ariyoshi, Y. Kimura, Y. Nakajima, Y. Takei,
M. Shimahara, T. Ueno and M. Suzuki1
Department of Dentistry & Oral Surgery, Division of
Medicine for Function & Morphology of Sensory Organs,
Osaka Medical College
1
Research Reactor Institute, Kyoto University
Introduction
In Boron neutron capture therapy (BNCT), selective andhighly concentrated boron accumulation in tumor cells is
important. A study was carried out in our division using
various liposomes ［1,2］.
Hyaluronan is a ligand of CD44 and RHAMM, which isexcessively expressed in tumor cells, and Hyaluronanconjugated PEG Liposome (HA PEG Liposome) is anactive targeting candidate for tumor cells.
In this study, BSH transport using HA PEG Liposomes
was investigated for oral squamous cancer cells, and theeffect of neutron irradiation was investigated by colony
formation assay.
Material and Methods
・Experiment on boron transport to tumor cells over time.
For oral squamous cancer cells, SAS was used. SAS wascultured in a serum-free media, and divided into groups
using Bare-BSH, PEG Liposomal BSH, and HAPEG
Liposomal BSH. The media was prepared so that eachgroup would contain a boron concentration of 30 ppm,
and each group was exposed to boron for 3 hours, 6hours, 12 hours and 24 hours. After the specified time,the media was removed, and cells were collected to count.
Then, nitric acid was added and ICP-AES was used tomeasure boron concentration.
・Experiment on neutron irradiation effect.
SAS was cultured in a serum-free medium, and divided
into groups using Bare-BSH, PEG Liposomal BSH andHA PEG Liposomal BSH, which were then adjusted to aboron concentration of 30 ppm. After exposure to boronfor 6 hours, cells were collected. Media which did notcontain boron was used to dilute the cells to 1×105
cells/mL, and set as samples of neutron irradiation. The
neutrons were irradiated for the pre-determined times.
After the neutrons were irradiated, cells were adjusted to
200 cells/petri dish and cultured for 7 days, and then anassay was carried out.

Results
・Boron transport to tumor cells over time
In the HA PEG Liposome group, boron concentration in
the cells increased over time. Moreover, increase in cellconcentration was hardly observed in the Bare-BSH andPEG Liposome groups. When the PEG Liposome and
HA PEG Liposome groups were compared, a significant
difference was observed at 3 hours (Fig. 1).
・Neutron irradiation effect
In the experimental results of the colony formation assay(Fig.2), the lowest number of surviving tumor cells wasobserved in the HA PEG Liposome group.
These experiment results suggest that for SAS, liposomes
combined with HA significantly transported more boron
into the cell, in comparison with Bare BSH. In the future,
we plan to investigate the effect of HA PEG Liposomes
on other oral squamous cell cancers and the possibility of
their use.

Fig.1 Boron Uptake

Fig.2 Surviving Fraction
REFERENCES:[1] T. Shimahara et al., Bulletin of the Osaka Medical
College. 55(2009): 21-29[2] Y. Kimura et al., Bulletin of the Osaka Medical College. 56(2010): 65-72

採択課題番号 23042
口腔癌に対する硼素中性子補足療法の基礎的研究
共同通常（京大・原子炉）鈴木 実 （大阪医科大学・口腔外科）伊藤雄一、島原政司、有吉靖則、
木村吉宏、中島世市郎、武井祐子

CO7-12

Boron Neutron Capture Therapy for Malignant Pleuritis

K. Ono, M. Suzuki, S. Masunaga, N. Kondo, M.
Narabayashi, Y. Sakurai, H. Tanaka, Y. Kinashi and A.
Maruhashi,
Research Reactor Institute, Kyoto University
INTRODUCTION: In2005, we have treated a patientwith left malignant plural mesothelioma (MPM) with
BNCT [1]. This attempt of BNCT for MPM was the firstcase in the world. Relief of clinical symptom was experienced at a few days after BNCT and regression of thetumor was observed on the computed tomography (CT) at
six months after BNCT. Although the patient died of local
tumor progression 10 months after BNCT, the survival
time was far over the expectation (less than 3 months) of
the attending physician. Based on this clinical experience
we are doing mul-ti-institutional clinical trials of BNCT
for MPM. It is prohibited to show a result of the trial in
the midway. But another type of malignant disease of thepleura, a malignant pleuritis of lung cancer, was treatedaccording to the similar protocol this year. Here we report
this clinical case.
2. METHOD and PATIENT
2-1.Treatment protocol
The whole ipsilateral lung should be regarded as clinical target volume(CTV). Since the maximum diameter ofcollimator available in BNCT was 25cm, the whole lung
was irradiated with posterior-upper and posterior-lower
beams in the first BNCT and with anterior-upper and anterior-lower beams in the second BNCT. The two BNCTs
were performed at 5-week interval. BPA(500 mg/kg)was
administered intravenously at the speed of200 mg/h in twohours just before irradiation and 100 mg/h during irradiation.
The Simulation Environment for Radiotherapy Applications (SERA) system, a currently available BNCT treatment planning system, was used for the calculation ofneutron fluence in each beam to deliver 5.0 Gy-eq to normal lung tissues in the ipsilateral lung.
2-2.Case report: A55-year-old man with right malignant
pleuritis from lung cancer was referred to our center for
further treatment with BNCT. He had received the surgery,chemotherapy of primary lung cancer (adenocarcinoma)
in 2003, and thereafter radiotherapy was applied to thesolitary metastasis in the lung in 2009. Limited pleurallesion and brain metastasis appeared in early 2010 and2011, respectively. They were treated by stereotactic radiotherapies. Brain lesion was controlled well by radiotherapy, but pleural lesion was gradually spreading to whole
pleura.
The patient received 18F-BPA PET, and 18F radioactivity

ratio between tumor and blood is > 2.0 (Tmax=5443Bq/ml
vs. Bmean=2719Bq/ml, Bmin.=2432Bq/ml). Based on
this examination data, we concluded BNCT is applicable.
First BNCT was performed on Dec.22 in 2011. Two anterior neutron beams irradiated the upper and lower portions of the ipsilateral lung. In second BNCT, the dorsal
portion of the ipsilateral lung was irradiated with two
posterior neutron beams on Jan.26 in2012.
Figures 1 show a B-10 level in the blood before andduring BNCT. The B-10 level was very stable at 22.5-24
ppm during neutron irradiation.
Fig.1

B-10
(ppm)

Time after the start of BPA administration (h)Dose volume histograms of the tumor, normal lung andliver were presented in figure 2 and 3. They show the firstand second BNCT, respectively. Large dose difference
between the tumor and normal organs, which is notachievable by X-ray therapy, was successfully obtained.
Fig. 2

Fig.3

The tumors in the pleura decreased in size and the condition of the patient is quite well. He has no respirationdifficulty, ex. short breath, cough and sputum. There is nochange in the normal lung by medical image examination
at present.
This new clinical experience in this year demonstrated
that if high enough accumulation ratio of BPA is expectedby 18F-BPA PET general malignant pleuritis is also agood indication of BNCT.
REFERENCES[1]M.Suzuki et al.Radiotherapy and Oncology88(2008)192–195.

採択課題番号 23052
中性子捕捉療法の臨床的研究
共同通常（京大・原子炉）小野公二、鈴木実、増永慎一郎、近藤夏子、木梨友子、櫻井良憲、田中浩基、丸橋晃

Intracellular Distribution of BSH-Appended Polyamine at Tumor Tissue ofColon 26 Carcinoma-Bearing Mice

CO7-13

M. Umano, Y. Zhu, J. Li, H. Azuma, T. Nagasaki, H.
Yanagie1, Y. Hattori2, M. Kirihata2, K. Ono3, S. Masunaga3 and Y. Sakurai3
Graduate School of Engineering, Osaka City University,
1
Graduate School of Engineering, The University of Tokyo, 2Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 3Research Reactor
Institute, Kyoto University
INTRODUCTION: Recently, we reported the evaluation of a polyion complex nanoparticle composed of
mercaptoundecahydrododecaborane-appendedhighmolecular weight polyamine (BPP, Fig. 1) as a novel
BNCT drug [1]. Since this polymeric 10B carrier has
anionic zeta-potential, polyion complexes with cationicpolymer (nEG-PLL, Fig.1) afford nanoparticles suitable
for safe and effective delivery into tumor tissues due toEnhanced Permeability and Retention (EPR) effect.
The neutron irradiation experiment by using polyion
complex containing BPP showed suppressive effect for
the tumor growth in colon 26 carcinoma-bearing mice.
Although the boron concentration at tumor tissue wasless than 10 ppm, the sufficient BNCT effect was observed. Here, we report the intracellular distribution offluorescently labeled BPP and BSH residues introduced
into the polymeric carrier using an immunostaining with
anti-BSH antibody.
EXPERIMENTS: For visualization of BPP, an aminogroup of BPP was covalently modified with
5-(and-6)-carboxy-X-rhodamine
succinimidyl
ester(Molecular Probes). Colon 26 cells (8 x 105 cells) weretransplanted into a left thigh of mice (4 weeks old
BALB/c, male). After 10 days of transplantation, therhodamine-labeled BPP complex solution was injected
O
S
N
O

O

H

C

N

OH

O
O

9

30

NH2

O

H

NH

N

O

HO

OH

N

C

H

O

C
NH2

OH

30-n

BSH-PEG-PLL (BPP)
NH2

OH

NH2
HO

C

N

O

H

O

30

OH

O

N

C

n

H

O

OH

30

nEG-PLL (n=9, 23)

Fig. 1. BSH-appended polyamine and based polyamines.

採択課題番号 23056

via tail vein at a dose of 4.0 mg 10B/kg (400 ppm of 10B
concentration; 200 µL). Twelve hours after injection, themice were euthanized. After the tumors were removed
and fixed in 4% formaldehyde, the tissues were freezed
in liquid nitrogen. Ten µm section were cut and treatedwith slow fade Gold antifade reagent with DAPI(Molecular Probes). Immunohistochemical characterization of BSH was carried out with pre-fluorescently labeled anti-BSH antibody by using Zenon mouse IgG Alexa Fluor 488-labeling kit (Molecular Probes).

Fig. 2. In vivo cellular uptake of BSH-modified polyamine by transplanted colon carcinoma. DAPI stained
nucleus (A), Rhodamine-labeled polyamine (B), andimmunostained BSH (C) were observed with confocal
fluorescent microscopy.
RESULTS AND DISCUSSION: BNCT using the
BPP complex was previously carried out with colon
26-bearing mice at left thigh. Tumor growth in micetreated with the BPP complex was obviously suppressed
after neutron irradiation. Under same conditions, injected
BPP-modified polyamine was fluorescently observed intumor cells. The colocalization of polyamine and BSH
residue indicates that BSH modification is stable duringin vivo experiment. Interestingly, perinuclear localization
was confirmed. It was reported that among proteins traveling between the cytoplasm and the nucleus, small proteins (up to 9 nm in diameter) can freely diffuse throughnuclear pore complex (NPC) [2]. As average particle sizeof BPP complex was estimated to be 91 nm, BPP complex could not pass NPC. Therefore, they remain at perinuclear regions. Morrison et al. reported that nuclear
localization of boron drug increased tumor-killing effectby BNCT [3]. The intracellular distribution in which
BSH is close to genome DNA might contribute higher
BNCT efficacy by BPP complex.
REFERENCE:[1] M. Umano et al., Appl. Radiat. Isot., 69 (2011)1765-1767.
[2] O. Keminer et al., Biophys J, 77 (1999) 217-228.
[3] G.H. Morrison et al., Biol. Cell, 74 (1992) 105-108.

ホウ素クラスター修飾ポリアミンの腫瘍集積性および中性子捕捉反応効率評価

共同通常（阪市大院・工）長﨑 健、東 秀紀、李 家暐、朱 宇翔、鹿子嶋祐太、林高一郎、河崎 陸（阪府大院・生命環境科）切畑光統、服部能英、門野尚之（京大・原子炉）小野公二、増永慎一郎

CO7-14 Boron Neutron Capture Therapy for Malignant Brain Tumors Using Epithermal Neutron

S. Miyatake, S. Kawabata, R. Hiranatsu, Y. Hirota T.
Kuroiwa, Y. Sakurai1, H. Tanaka1, A. Maruhashi 1, M.
Suzuki1, N. Kondo, S.. Masunaga1, Y. Kinashi1 and K.
Ono1.

Fig. 1.

Department of Neurosurgery, Osaka Medical College
1
Research Reactor Institute, Kyoto University

INTRODUCTION:
Between March 2010 and
Februarry 2011, we applied BNCT using epithermal
neutron for1) newly diagnosed GBM (8 cases),2) recurrent malignant gliomas (7 cases)
3) malignant meninigiomas (MM)(4 cases).
These BNCT were appied in KUR (15 cases) and JRR4 (4cases).
Methods
BNCT using epithermal beam were applied in 3 different
categories of maliganant brain tumors as menitioned
above.
1) Newly diagnosed GBM patients were treated as Phase
II Study of Boron Neutron Capture Therapy Followed By
Radiotherapy and Concurrent and Adjuvant
Temozolomide in Patients With Newly Diagnosed
Glioblastoma Multiforme (TRIBRAIN0902).
2) All recurrent malignant gliomas treated by BNCT were
TMZ-refractory cases.
3) All malignant meningiomas were recuurent cases after
different radiation modalities.
RESULTS and Discussion1)TRIBRAIN 0902 is now in progress, Thereafter
results will be opened a cupple of years later..
2) For recurrent malignant gliomas, 2 cases were dead by
CSF dissemination and 5 cases are still alive. A
representative case of maliognnant gliomas is depicted inFig. 1 and 2.. Radiation necrosis should be a serious
problem in the recurrent cases treated by BNCT. We
applied bevacizumab for the treatement of symptomaticradiation necrosis. {1,2}3) We depicted the representative case of MM in Fig.3.
Original tumors are controlled well, but unfortunatedly a
new lesion was noticed in the right frontal base (out ofradiation field).
REFERENCES:.
[1] Furuse M, Kawabata S, Kuroiwa T, Miyatake SI. :Repeated treatments with bevacizumab for recurrent
radiation necrosis in patients with malignant brain
tumors: a report of 2 cases.
J Neurooncol. 2011
May;102(3):471-5.

採択課題番号 23060

Fig. 2

Fig. 3[2] Nonoguchi ＮMiyatake SI, Fukumoto M, Furuse M,
Hiramatsu R, Kawabata S, Kuroiwa T, and Ono K.: The
distributionofvascularendothelial
growthfactor-producing cells in clinical radiation necrosis of thebrain. - Pathological consideration of their potential roles-” in press in J Neuro-Oncol

熱外中性子を用いた悪性脳腫瘍に対する非開頭中性子捕捉療法の
共同通常
臨床的研究
（大医大・脳外）宮武伸一、黒岩敏彦、川端信司、平松亮（京大原子炉）小野公二、増永慎一郎、鈴木 実、
近藤夏子、田中浩基、櫻井良憲, 丸橋 晃

CO7-15High LET-radiation Can Overcome Radioresistance of Glioma Stem-Like Cells
Which are Radioresistant to Low LET-radiation

Y. Hirota, S. Masunaga1, N. Kondo1, S. Kawabata2,
A. Fujimori 3, K. Ono1, T. Kuroiwa2, S. Miyatake2
Graduate School of Neurosurgery, Osaka Medical College
1
Research Reactor Institute, Kyoto University
2
Department of Neurosurgery, Osaka Medical College
3
National Institute of Radiological science, Heavy-ion
Radiobiology Research group, Research Center for
Charged Particle Therapy
PURPOSE:
Ionizing radiation is applied as the standard treatment forglioblastoma multiforme (GBM). However, radiotherapy
remains only palliative because of existence of glioma
stem cells (GSC), which are regarded as highly radio-resistant to low linear energy transfer (LET) photons[1, 2]. On the other hand, we have applied boron
neutron capture therapy (BNCT) for GBM. This is aunique tumor-selective particle radiotherapy using neutron irradiation, especially thermal neutron irradiation.
Boron-10 (10B) releases alpha (4He) and 7Li particles by
10
B(n,α)7Li reaction. The key players of anti-tumor effects in BNCT are these high linear energy transfer (LET)
particles. With BNCT, good results have been achievedfor patients with GBM and recurrent malignant glioma[3,4]. Here we analyzed whether high LET particles canovercome the radio-resistance of GSC or not.
MATERIALS AND METHODS:
Glioma stem-like cells (GSLCs) were induced from
GBM cell lines A172 and U251 MG in stem cell-culture
medium[3]. The phenotype of these GSLCs and wild typecell lines were confirmed by western blot analysis using
stem cell markers. These cells were irradiated with 60Co
gamma rays or neutron beams. Radio-sensitivity wasassessed in terms of clonogenicity using colony forming
assay[4] and the number of DNA double strand breaks(DSBs) using histone gamma-H2AX foci detection assay[5, 6].

RESULTS:
In stem cell-cultured medium, GSLCs could form neurosphere-like cell and expressed neural stem cell markers
more frequently. GSLCs were radio-resistant to gamma-rays in comparison with parental cultured cell lines,
but neutron beams could overcome the resistance. Twenty-four hours after irradiation with gamma-rays, thenumber of gamma-H2AX foci in GSLCs was significantly less than that of parental cells, while there was noapparent difference in the number of these foci between
GSLCs and parental cultured cell lines following neutron
beam irradiation.
CONCLUSION:
Neutron beams can induce elastic scattering and nitrogenneutron capture reaction, and produce proton particle(H+). This particle is high LET radiation and it could
overcome radioresistance of GSLCs with unrepairable
DSBs. Therefore, High LET radiation therapy has apromising potential to overcome X-ray resistance of glioblastoma in clinical setting.

REFERENCES:[1]S. Bao et al, Nature 444 (2006) 756-760.
[2]C. Charn-jung et al, Biochemical and Biophysical Research Communications, 380 (2009) 236-242.
[3]S. Miyatake et al, Journal of Neuro-oncology, 91(2009) 199-206[4]S. Kawabata et al, Journal of Radiation Research, 50(2009) 51-60[5]Q. Lei et al, Cancer Letters 279 (2009) 13-21[6]A. Maria et al, International Journal of Radiation Oncology Biology Physics, 79 (2011) 262-268[7]A. Osma et al, International Journal of Radiation Oncology Biology Physics, 75 (2009) 1216-1225[8]Y. Kinashi et al, Radiation Oncology, (2011) 6:106

採択課題番号 23061
新規硼素化合物の有用性と腫瘍幹細胞の克服（大阪医大）宮武伸一、弘田祐己、川端信司、平松亮、米田隆（岡山大）松井秀樹、道上宏之、森亜希子、秋田直樹（熊本大）富澤一仁（京大原子炉）小野公二

共同通常

CO7-16

The 7umor &ontrol (ffect by the Sonoporation in BNCT

N. Yamamoto, Y. Yura, I. Kato, Y. Fujita1, M. Okamoto2, K. Ono3, A. Maruhasi3, M. Suzuki3 , S. Masunaga3
Y. Sakurai3 and H.Tanaka3
Graduate School of dentistry, Osaka University
1
Dentistry-Stomatology, Rinku General Medical Center2
Graduate School of Pharmacy, Musasino University
3
Research Reactor Institute, Kyoto University

irradiation, the tumor control effect was seen as compared with control. In the group which carried out neutron irradiation after injected BPA and performed
sonoporation、the individual for which a tumor disappears existed (Fig. 1).The average survival period of thegroup extended.

INTRODUCTION: The sonoporation which is theultrasonic irradiation of a low output is one of the methods of introducing a medicine and a gene into a cell. Useof the microbubble which is an ultrasonic contrast agent
will raise introductory efficiency.[1,2] We have reported
that the boron concentration in tumor cells is increase byuseing sonoporation. We experimented by carrying out
nude-mouse use for the purpose of curative effect verification of BNCT in human oral squamous cell carcinoma(SAS).


EXPERIMENTS: We transplanted SAS of the thighsubcutaneous part of the nude mouse (bulb/c 4week), andthe major axis used what became not less than 5 mm. We
carried out intraperitoneal injection of the BPA, and injected 0.1 ml of microbubble into the tumor 2 hours afterward. We performed ultrasonic irradiation immediately and irradiated with neutron ray at KUR.

Fig.1. Change of tumor volume by BNCT (S: sonoporation
with microbubble, N:neutron irradiated).
REFERENCES:[1]R.Suzuki et al.,J Cnotrol Release.,125(2008)137-144[2]Y.Watanabe et al.,Cancer Sci.,99(2008)2525-31


RESULTS: Into the group which carried out neutron

採択課題番号23063

口腔悪性腫瘍におけるホウ素中性子捕捉療法に関する基礎研究

共同通常（京大・原子炉） 小野公二、丸橋晃、鈴木実、増永慎一郎、櫻井良憲、田中浩基（阪大院・歯）加藤逸郎、由良義明、山本直典、（武蔵野大・薬）岡本正人（泉佐野病院・口外）藤田祐生
共同通常CO7-17

Clinical Studies on BNCT for 4 Cases of Head and Neck Cancer

I. Kato1, N. Yamamoto2, Y. Fujita3, M. Ohmae4, M.
Suzuki1, S. Masunaga1, M. Nakazawa, S. Iwai, A.
Maruhashi1, Y. Imahori6, M. Kirihata7 and K. Ono1
1

Dept. of Oral &Max.fac. Surg. II, Grad. Sch. of Dent.,
Osaka Univ.,
2
Dept. of Oral &Max.fac. Surg., Saiseikai-Senri Hpsp.,
3
Dept. of Oral &Max.fac.Surg., Higashiosaka General
Hosp.,
4
Oral &Max. fac. Surg., Rinku,General Med. Center,
5
Dept. of Oral &Max.fac. Surg., Rinku General Medical Center,
6
CICS Inc., 7Graduate School of Agriculture and Life
Science, Osaka Prefecture University
INTRODUCTION: We have first reported that six patients with head and neck malignancies (HNM) had beentreated with BNCT [1]. We summarized 4 patients with
HNM who had treated with BNCT at KUR in last year in
Table 1. Moreover we also report here long term (morethan 5-year) clinical outcomes of our 26 patients withrecurrent HNM treated with BNCT [2].
PURPOSES: The purpose of this study was to estimate safety and effectiveness of BNCT for patientswith advanced/ recurrent HNM for which there wereno other treatment options.
RESULTS: All cases are advanced such as 15 out of
26 patients (58%) had developed regional lymph nodemetastases. Distant metastases were developed in 6
cases during treatment. (1)10B concentration of tumor/normal tissue ratios (T/N ratio) of FBPA-PET
studies were SCC: 1.8-5.7, sarcoma: 2.5-4.0, parotid
tumor: 2.5-3.7. (1) Regression rates were CR: 12cases(46%), PR: 10cases (39%), PD: 3cases (12%), NE (notevaluated):1case. Response rate was 85%. (2) Mean
Survival time was 33.6 months. 2-year overall survival
rate (OS) and 6-year OS were 37.0% and 31.7%, respectively. (3)BNCT improved QOL, PS and survival
periods. (4)Survival periods after BNCT were 1-84
months. (5) Adverse events were brain necrosis, osteomyelitis and transient mucositis and alopecia and soon [2]. After 5-year interval for renewal of low enriched uranium-KUR BNCT was restarted in 2010. We
had treated 4 times of BNCT for 4 patients last year.

Case 1: A 60 year-old female with op. upper gingival
carcinoma (rT0N3M0, SCC) had got maxillectomy in2009. About half and a year after the operation, as she
had developed lt-cervical metastatic lymph nodes, she
had go the neck dissection. FBPA-PET study revealed10
B concentration of T/B ratio was 2.6. Then she has
been treated with BPA (500mg/kg) mediated BNCT at
KUR in December 2011 and she has been disease freesurvival for so far 6-month.
Case 3: A 63-year old woman had been to the department of oral and maxillofacial surgery of a municipal
hospital with chief complaint of swelling and pain at
the left temporal mandible joint (TMJ) since July 2011
and she was diagnosed as temporal mandibular disorder (TMD) type I. She was prescribed indomethacin
cream and Ternelin, but the swelling of L-TMJ become worse. She had referred to our hospital. MR image revealed that the swelling was caused by masslesion which was T2-high signal and with calcification.
She was diagnosed as benign tumor at TMJ and thelesion was incised at the municipal hospital in November. Histopathological diagnosis was “Osteogenic
sarcoma, chondroblasitc type ”
We consulted diagnosis and treatment method in thiscase with Prof. Yoshikawa at the department of orthopedics, medicine, Osaka University. and he suggested
us that in this case, heavy particle therapy including
BNCT was indication but multi drug chemotherapy
was contraindication for her age. FBPA-PET study
resulted that T/N ratio=2.3、T/B ratio=2.2. She treated
BNCT in January 26, 2012. Then she has been disease
free survival for so far 5-month.
CONCLUSIONS: BNCT was effective in 22 out of
26 patients and safe in all case. BNCT represents a
new and promising treatment even for far-advanced
patients with a huge HNM.
REFERENCES:[1] Kato, I. et al., Applied Radiation and Isotopes
2004; 61:1069-1073[2] Barth R, et al., Radiation Oncology ; in press

Table 1. Treatment Summary of 4 Cases
Case
No.
1
2
3
4

Pt‘s Initial(Age)
W・Y (60)
F・M（79）
Y・Y (63)
O・Y(34)

Clinical Diag.
(Histopathol. Diag.)
Op.OKK, Lt-LN meta(SCC)
bds-mandible Ca.(ACC)
L-TMJ Tumor (Osteosarcoma)
0p.OKT (ACC)

10B-conc.
Blood(ppm
)
20.0
28.0
32.0
22.7

T/B
ratio
2.6
2.2
2.2
2.9

T-max of thermal neutron (D)Fluence
History of(E+11n/cm-2)
RT: (Gy)
18.0
63.3
―
23.0
―
25.0
15.0

Total-RBE-Dose Eq (Gy-Eq)
T-Peak Gy-Eq
T-deepest
Skin/Mu
Gy-E
cosa
25.0(4cm)
28.3（2.2 ㎝）
4.9/12.0
21.0(6cm)
42.0（2.4 ㎝）
6.2/13.0
28.0(4cm)
52.0（2.4 ㎝）
/15.0
23.0
29.0
4/14

Irradiationtime(min.)(May , 2012)% Reduction (Period)
Prognosis (Survival)

87
61
60
44

CR(5M)・Alive(5M)(4M)・Alive
CR（4M）Alive(（4M）
CR（3M）Alive(（3M）

採択課題番号 23064
頭頸部悪性腫瘍におけるホウ素中性子捕捉療法の臨床的研究
共同通常（阪大 2 口外）加藤逸郎、岩井聡一、墨哲郎、中澤光博、由良義明（阪大工）村田勲（慶大先端医研）岡本正人（りんくう医療セ）大前政利（済生会千里）道澤雅裕（東大阪総合）千足浩久（市立池田）大西徹郎（田中クリニック）田中善（京大炉）田中浩基、鈴木 実、櫻井良憲、増永慎一郎、
丸橋晃、小野公二

CO7-18
Basic
Cell Survival Analysis of New Boron and Gadolinium CompoundsIRU1HXWURQ&DSWXUHTherapy
K Nakai, M Shirakawa, Y Yamamoto, T Yamamoto, F
Yoshida, Y Uemae, E Sato, H Tanaka1, Y Sakurai1 and S.
Masunaga1
Graduate School of Comprehensive Human Science, Department of Neurosurgery, Faculty of Medicine,University of Tsukuba
1
Research Reactor Institute, Kyoto University
INTRODUCTION:In Neutron Capture Therapy(NCT) , the therapeutic effect of the boron compound isbased on alpha particles produced by the B(n,α) reaction.
On the other hand, that of gadolinium compound isgamma rays derived from the Gd (n,γ) reaction. There isno clinical trial using Gd-NCT because the long pathway
of gamma ray may cause the radiation damage of normal
surrounding tissue and the uptake and washout of
Gd-DTPA within the tumor tissue is only several minutes.
In a previous study, we analyzed Gd-containing porphyrin[1], and combined boron and gadolinium are warranted to prove the efficacy. we select and analyzed the neutron capture effect for the cancer cell line with the boronated liposome and Gadolinium containing nanoparticles
to investigate this possibility.
EXPERIMENTS: Colony assay with CT26 (undifferentiated colon carcinoma cell line) and C6 (rat glioma
cell line) were performed.
For the gadolinium targeting compounds, Gd contaioning
nanoparticle was used in this study. Gd containing nanoparticle was diluted to reach a final gadolinium concentration of 0, 20 and 40 ppm.
The boronated liposomes were prepared by thelipids (table 1). The lipid mixture prepared using
the constant ratio was dissolved in organic solvent. It was prepared by the conventional lipid-film method. The resulting liposomes were
extruded through polycarbonate membrane using
an extruder, yielding the modified liposome.

PBL5%
PBL10%
PEG

DSPC
47.5
45
47.5

Cho
47.5
45
47.5

PEG-B-Lip
5
10

Fig.1 The results of colony formation assay (Gd nanoparticle). The survival fraction was plotted as a functionof neutron irradiation time. ◆40ppm Gd/MEM, ■20ppmGd/MEM, ＊neutron only.

DSPE-PEG
5

Table1 composition of liposomes.
DSPC: α-phosphatidylcholine distearoyl Cho: cholesterol PEG: polyethylene glycol DSPE: α-distearoylphosphatidylethanolamine
Thermal neutron irradiation was performed at Kyoto
University Research Reactor (Osaka, Japan). The irradiation times were 0, 15, 30 and 45 minutes.

採択課題番号23078

RESULTS: As shown in Fig. 1, Gd nano-particle andBoronated liposomes have some enhancement effects.
further investigation of in vivo and in vitro experiments
are expected.

Fig.2 The result of colony formation assay (boronatedliposome and BSH). PBL modified 5% liposome
group (◆) showed same cytocidal effect the BSH
5% group (■) and lower cytocidal effect than the
BSH 10% group (▲).
REFERENCES:[1] A MAtsumura et al., J. Exp. Cancer Res.,24, 1, (2005)
93-98.

新規化合物および細胞内濃度変調による共同通常
中性子捕捉反応による細胞生存試験（筑波大学・人総）中井啓、山本哲哉、吉田文代、白川真、山本陽平、上前洋二、佐藤英介（京大・原子炉）田中浩基、櫻井良憲、増永慎一郎