dds c ,bc ]^ · 17 % cell growth dmbl 100.00 ppm dmbl 33.33 ppm dmbl 11.11 ppm control dmbl 3.70...

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DDS BNCT 2 DDS T 50% 40 ppm T/B>10 100%

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Page 1: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

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Page 2: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

3

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��FGHNOP���w�����FGHxyr��$�������� �����_h¡¢£¤¥�_`r¦§

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Page 3: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

5

FFGH�iFG�t��¾¿IJKLM=�FGHNOP

ï��*�ô�ïï*�ô�tFGHgRD3¾¿IJKLM=�FGHNOP

4-Boronophenylalanine Ethylester(BPA-OEt)

4-Boronobenzylcysteine Ethylester(BBC-OEt)

NH N

HNN

NHHN O O

HO NO

O

NaOONa

O

10BHO OH

10BHOOH

NH N

HNN

NHHN O O

HO NO

O

NaOONa

O S

10BHO OH

S

10BHOOHEC-028 EC-031

10B

COOEt

HO

OH

NH2 10B

SHO

OH

COOEt

NH2

��.� P. 98B-1:FGHIJKLM=���

6

FFGH.�� Jß�]±FGH�i�*�J�­=t��¾¿IJKLM=�FGHNOP

IJKLM={O��i�J�� J�tgR¾¿FGHNOP

4-Carboran-1-ylmethoxybenzoic acid (CMBA)

OC

HC

OH

O

: nBH

NH N

HNN

NN O O

HO NOHN

O

ONa

O

ONaOONa

O

O

NaO

O C

HC

: nBH

EC-032

��.� P. 100

Page 4: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

7

��67ië�kFGHIJKLM=EC031, EC032���È�

��.� P. 103

8������������ ���

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� � ����������������

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FGHIJKLM=EC031(b�ue), EC032(red) �67wxy����N

67wFGHWX���g/108 cell)�

�>�67�U251:73.8 ppm

��.� P. 104

��.� P. 105

Page 5: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

9

FFGHNOPQRHVJ-Envelope (HVJ-E) Vector���

���?@ABr� !Í

B-2:FGHWX�DDSÏD���

ñ#$%&'()**

����Pilot plant����

���� !

�+ �,+STOijk-.gR

T 67YZ67YZ67

-bc�/�N�

04211i,���¿¼�����i¤Â23

-Nucleocapsid, -Polymerase

HVJ-E (Empty vector)Wild type HVJ HVJ-E (+BSH)HN

F

Nucleocapsid(RNA, NP)

M

HVJPolymerase

(P, L)

45V

CÆ6/�DC¡7

SH

BSHSH

BSH

��.� P.127

HVJ-E��89:ê;<=>?@A�4BCÉ(�DEêFGàH�

10

Inactivation BSHLipid mixture

HVJ-liposome

Liposome with boronated compound

HVJ

SH

BSH

HVJ-liposome�I��89:ê9JM��4BCÉ(�DEêKaneda, Y., Dzau, VJ�

Page 6: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

11

0

20

40

60

10 12 14 16

Added B (ug)Cel

l B c

once

ntra

tion

(ug

/g)

CT26 LM8

BSH delivery to cancer cells by HVJ-liposome

KL�rs67i40 ppmtMNkFGHxy�[\r!Í(HVJ-E�OPQ�

BSH�RS���TOU 0.12 – 0.18 Vg/107cells (4.8-7.2ppm)

��.� P. 131

12

W#$�X'ï'(�&()&YX%YZ�&

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Zfgf

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BSHQRHVJ-E�íØ���80 �g 10B/HVJ-E/tumor �

CT26��FGHxy

BSH �80 �g 10B��ìi�íØ��wxy:12-27 ppm

�xyê86.4 ppm(75.8 ppm at 1hr, 93.5 ppm at 1.5 hr)

��.� P. 133

Page 7: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

13

ï*��c'j�X�&'(�&()&YX%YZ�&

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BSH�íØ��!�Ý�xyê0.6 – 1.1 ppm2000 �g 10B/body �lmw��24�n!�Ý�xyê0.6 ppm

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14

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Page 8: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

15

FFGHMIUB����

double tail

H. Nakamura, Y. Miyajima, T. Takei, S. Kasaoka, K. Maruyama, Chem. Commun. (Hot paper) 2004, 1910.

hydrophilic part

rs��J�­=tu�À��XÌO4qvæwm濼xyai�æ ÓÊz¿FGH@­�AB�

ý{E5�| Ó¿rh�ö÷�| Óf�}¿~ý{E5�| Ó¿rh�ö÷�| Óf�}¿~FGHxy�FGHxy�7.2 mg/kg7.2 mg/kgr�����Æhr�����Æh14.4mg/kg14.4mg/kg������r| Ó¿~������r| Ó¿~

NaH

O OOH

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16

n = 16: Stearoyln = 14: Parmitoyln = 12: Myristoyl

O NH

O

OO

O

H3C(H2C)n

O

H3C(H2C)n

S

2-O

M=�B����R

DMCBL (n = 12)DPCBL (n = 14)DSCBL (n = 16)

OS

O

OO

O

H3C(H2C)n

O

H3C(H2C)n

2-

DMBL (n = 12)DPBL (n = 14)DSBL (n = 16)

O Y

OS

2-

BC (Y = CH2)BCC (Y = NHCOCH2)

����FGHMIUB��$���

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BB1212HH1212SH (BSH)SH (BSH)

SH

2

--

Liposomes consist of phospholipids and cholesterol

(9�ê2006-186907h2006-275367h2006-259290�

��.� P. 108

Page 9: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

17

% c

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h

DM

BL

100.

00 p

pmD

MB

L 33

.33

ppm

DM

BL

11.1

1 pp

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MB

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3.70

ppm

DM

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1.23

ppm

cont

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.33

ppm

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L 11

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70 p

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1.23

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DSB

L

100.

00 p

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33

.33

ppm

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70 p

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ppm

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.70

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BL

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.00

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BL

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3.33

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1.23

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DSC

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100

.00

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DSC

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3

3.33

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3.70

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ppm

BSH

100.

00 p

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SH

33

.33

ppm

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11.1

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SH

3.70

ppm

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1.

23 p

pm

B(O

H)3

100.

00 p

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(OH

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.33

ppm

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11.1

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70 p

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23 p

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BC

100.

00 p

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��.� P.110

18

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Mice died.

Tumor disappeared!

control

DPBL(50%) Liposomes(5 mg10B/kg)

BSH (57 mg10B/kg)

PEGPEG--DPBL(50%)DPBL(50%)FGHMIUB��FGHMIUB������jÐ�xy�������rö÷jÐ�xy�������rö÷

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Page 10: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

19

FGHMIUB��ÙG�ilmk��ëjù����n!���wFGH���úxy

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20

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à3�È�hXE�È�hàá�È��{5h25% or 50% DSBL (10-20 mg B/kg)tin vivo� ¡mkq3i¢3

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DSBLê 50% 25%wê13.4ppm 22.6ppmÝÞ�ê0.76ppm 37.6ppm

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Page 11: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

21

,f,

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+,,f,

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Tumor shrinkage

PEG

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22

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10mg10B/kg >42 days

0³´{ 8 days

5mg10B/kg

DSBL-FGHMIUB��XE�hHVJ-FGHMIUB��]^E5�¡D

��11��FGHMIUB�����FGHMIUB���ý{E5ý{E5�É~�É~�ö÷�| Óf�}¿~�ö÷�| Óf�}¿~

FGHxy�FGHxy�7.2 mg/kg7.2 mg/kgr�����Æhr�����Æh14.4mg/kg14.4mg/kg������r| Ó¿~������r| Ó¿~

Bioconjugate Chem. 2006, 17, 1310.

��.� P. 114

Page 12: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

23

FGHNOP�wxy3T/Bß

HVJ-E(BSH)��ìi�íØ��� 32mg 10B/kg �ÙG�{õ,ê�xyê86.4 ppm, ÝÞ�xyê0.5 ppmÙG�µ0q�,ê�xyê135 ppm, ÝÞ�xyê0.5 ppmwFGHxy40ppmpq, T/B>10tu4BSH�ÙG�{õ,ìi�íØ��wxy:12-27 ppmhÝÞ�xyê0.6-1.1 ppm

�2��FGHMIUB��ÙG�{õ, ¶Ji�áî��20mg 10B/kg ��24�n!�wxyê13.4 ppm, T/B=17.6T/B>10tu4

��?@AB�ÙG�{õ, ¶Ji���� >40ppm, T/B>10 �u4�!Í��1���FGHMIUB�(Bioconjugate Chem. 17, 1314-20, 2006)7.2mg 10B/kg ��24�n!�wxyê23.1 ppm, T/B=2.3BSHQRTf-PEG-liposome (J. Control. Release 98, 195-207, 2004)35 mg 10B/kg ��72�n!�wxyê33.1ppm, T/B=6.0BSHQRPEG-liposome (J. Control. Release 98, 197-205, 2004)35 mg 10B/kg ��72�n!�wxyê24.3ppm, T/B=2.5

24

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Page 13: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

25

�4$MIUB��Ë100 nm��XV$È�3��Ì4�ÍÎÏÐ/�VÑÒ�âÄ�IRSI� � kirrad / kcont �Ó�íÔ

q�Ô¿Ð�Ì4ßrÕO/�V�ÖÓwmæ~

XChol

X DLO

PC XDSPC

0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1

0

0.1

0.2

0.3

0.4

0.5

0.7

0.6

0.8

0.9

1

1

0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0

OO

OO O P O

O O-N+

H H

Dilinoleoyl phosphatidylcholine(DLOPC)

OO

OO O P O

O O-N+

Distearoyl phosphatidylcholine(DSPC)

HO

H

H

H

Cholesterol (Chol)

�æ¿��� ×Ô�à3

MIUB�×4�ÍØÙ

Akamatsu, K., Effect of ionizing radiation on membrane permeability of liposome: the dependency of lipid composition, concentration, and radiation dose/dose-rate, Chem. Phys. Lipids, to be published.

��.� P. 116

C-1: �����ÁÚtÀ�¾¿-,D�Ü=>«BJMMB����

26

0

20

40

60

80

100

0 5 10 15 20 25Time for incubation (h)

Effl

ux (%

)

Û�����ijkFGH��WMIUB� �DLOPC: DSPC: BC: Chol = 20: 20: 20: 40 �Ó�høùDLOPC: DSBL: Chol = 20: 20: 60 �Ü��/ M=�ÝÞßàÞ�pH 7.4�� �-

,D�J�ª­á=�/0����Ü=>«BJ�£âãhä�åæ�~MIUB�çèÞxyê),'$éh],ïxyê],,,'hh$~

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«­ê«ëì3Chol�{O�íë«+êíëî­Q�«ëìWXïð��

��.� P. 123

Page 14: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

27

MIUB�w�iïðñ3(%Xj�h*%YZ&tQRmk

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2 Na+

~ pH 11

28

ïððïQRMIUB��Û���ÑÒ�tO·v Û���K­�@�ê 1.64 x 109 n/cm2/s

v ���nê 4�n

v ��É��xyê ~ 10 mM

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Page 16: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

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Page 17: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

33

siRNA

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34

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Page 18: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

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Page 19: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

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38

Page 20: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

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Page 21: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

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Page 22: DDS C ,bc ]^ · 17 % cell growth DMBL 100.00 ppm DMBL 33.33 ppm DMBL 11.11 ppm control DMBL 3.70 ppm DMBL 1.23 ppm DPBL 100.00 ppm DPBL 33.33 ppm DPBL 11.11 ppm DPBL 3.70 ppmDPBL

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