Nagasaki University

1
??20?? ???????? ?4? ??20?6?4?
???????????????????DDS
???? ???????????? ??????? ?????? ???
? ? ? ? Koyo Nishida, Ph.D. koyo-n_at_nagasaki-u.ac.j
p
Nagasaki University Graduate School of Biomedical
Sciences
http//en.beijing2008.cn/
2
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• ??????? ?????1986-1991
• ????????????????????(????)
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• ???? ???????????????1991-
• ??????????????????????????DDS???
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DDS (??????????)
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lt ??????????????????? (2005???) ?? gt
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24L/? ???? ?5-6 L/min ??? ?60-70?/min ?????? 1
?????????????? 40 ???????
10000 L/day
http//www.kyushin.co.jp/
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EPR??
Enhanced permeability and retention (EPR) effect
????????????????? ???????? ??????????????? ???????
????????????????????????? ??????????????
????i.v. 24 h??????
Maeda et al, Adv. Drug Deliv. Rev. 46169-185
(2001)
http//www.nanocarrier.co.jp/
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blood-brain barrier ?????????????????????????????
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????????BBB???????????????????????????????????????
(tight junction)????????????
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??osmotic opening?????????????????????????????????
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?? ????DDS ???
??????????BBB????
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???????in vitro??in vivo???
? in vivo??? ???????? ??????????? ????????????
? in situ(????) ????????? ????????? ??????? ?
?????????
Liver
Rat (in vivo)
????????
Hepatocytes
?in vitro(?????) ?????????? ????????? ????????
? in vivo????????
16
????????????
??????????
??????????????????????????????????(Induced
Hypertention ChemotherapyIHC)
Nagamitsu et al, DDS, 22, 510, 2007
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????????????? http//med2.astellas.jp/jp/
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kidney surface
i.a.(bolus, constant)
i.v.
Well-stirred model
Nishida et al, J. Drug Target., 13, 215, 2005
18
????????????
kidney surface
??? ? Applied kidney ? Non-applied kidney ?
plasma
i.v.
??????????? - Applied kidney ??? Non-applied
kidney -- plasma
Phenolsulfonphthalein????????????
Nishida et al, J. Drug Target., 13, 215, 2005
19
?????????????
i.a.(constant) 4.2 µg/min for 240 min (1mg)
i.a.(bolus) 1 mg
??????????? - Applied kidney ??? Non-applied
kidney -- plasma
???????????
Phenolsulfonphthalein????????????
Nishida et al, J. Drug Target., 13, 215, 2005
20
????????
????????????????????

• ????
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• 5-FU?, TS-1, ??????,(????)
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• 5-FU??, (??????)
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• 5-FU???????

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http//ganjoho.ncc.go.jp/public/
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????????? http//www.hgh.or.jp/
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  ????.????,???????????????????????

??????????? http//cancer-words.seesaa.net/
24
????????????

• ???? IntraDose
• 5-FU or cisplatin/epinephrine????? Lancet 4 199
  (2003)
• ???? OncoGel
• paclitaxel ???Anticancer Drugs 18 283 (2007)
• Implant
• doxorubicin ?? J. Biomed. Mater. Res. 81A 205
  (2007)
• ?????
• 5-FU etc ?? Lung Cancer, in press (2008)
• ??????
• cucurbitacin ??? Int. J. Pharm. 347 118 (2008)
• ??????
• doxorubicin ??? Eur. J. Pharm. Biopharm. 65 300
  (2007)

??????? ?????? ????????? ???????? ??????
J. Pharm. Sci., 97 1681 (2008)
25
??????????????

1. ?????????
2. ?????????
3. ?????????
4. ?????????
5. ????????????

???? ? ???????????????????
Cancer Sci. 98 11 (2007)
26
??????????
Int. J. Gynecol. Cancer, 17 1 (2007)
Clin. J. Oncol. Nursing, 11 881 (2007)
27
?????????????
Int. J. Gynecol. Cancer, 17 1 (2007)
Cancer Res. 49 3380 (1989)
28
????????????
ASCO 2006 ????? ?????????????????????????????????
???????????5???????????? ??????III??????????????(
SCOG)?? SGC3?
Int. J. Gynecol. Cancer, 17 1 (2007)
http//www.medical-tribune.jp/congress/asco2006/
29
?????DDS?????

• ????????? MMC ??????? Drug Delivery System 2 134
  (1987)
• ??? MMC ???? Lancet 14 339 (1992)
• ??????? carboplatin ???????? Drug Delivery System
  14 185 (1999)
• ???????? cisplatin ????? J. Control. Rel. 80 295
  (2002)
• PEG????? doxorubicin ????? Toxicol. Lett. 116
  51 (2000)

Drug Delivery System, 22 522 (2007)
Doxil??????????PEG?????
30
?????????????????(???)
????????? NCIhttp//www.cancer.gov/
NHK???????
??????(230)
??????(????ET)
?????(236)
http//www.etclinica.com/ http//www.tbs.co.jp/yum
etobi/backnumber/20070121.html
31
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32
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33
???????????1
Pingpank, ???, Alexander et al, J. Clin. Oncol.,
23, 3465, 2005
34
???????????2
Pingpank et al, J. Clin. Oncol., 23, 3465, 2005
35
???????????3
??????
Pingpank et al, J. Clin. Oncol., 23, 3465, 2005
Grover A, Alexander HR Jr. The past decade of
experience with isolated hepatic perfusion.
Oncologist, 9, 653, 2004.
36
?????????1
Scheme of two-compartment physiological model
lti.v. constant infusiongt
plasma
Vs
Cs
systemic
Qs
local
???? k0????
target organ
Vt
lti.a. constant infusiongt
Qt
Ct
Et
rest
CLs
Daemen et al, Adv. Drug Deliv. Rev., 6, 1, 1991
37
?????????2
?????????????????? ????????? ???????????????????
? ????????????
ltsystemic advantagegt
ltregional advantagegt
Daemen et al, Trends Pharmacol. Sci., 9, 138-41,
1988
38
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????, 123 681-689 (2003)
40
??????
Development of drug delivery system to achieve
site-specific delivery or prolonged retention in
the circulation has attracted attention, because
new types of drugs are expected to be created
with advance in life science and biotechnology
such as human genome project. We have tried to
develop a new administration route for drug
targeting to the liver, since normal drug
administration by intravenous and oral route have
difficulty in achieving a local site of action in
the liver. Although the direct way of
application to the liver surface should yield
local drug distribution, the drug absorption from
the liver surface had not been reported in the
literature. Therefore, we have analyzed the
absorption mechanism of several organic anions
and dextrans with different molecular weights as
model drugs, after application to the rat liver
surface in vivo, employing a cylindrical
diffusion cell. Every compound appeared
gradually in the plasma, followed by excretion
into the bile and/or urine, indicating
possibility of drug absorption from the liver
surface. A specific transport system might not
be involved in the absorption process from the
liver surface, because the effect of dose and
transport inhibitors on the absorption was not
recognized. In addition, molecular weight was
found to be a determinant factor of absorption
from the liver surface. The targeting efficacy
to the applied region in the liver was
considerably enhanced by application to the liver
surface, as compared to intravenous
administration. Moreover, we have obtained
important physicochemical and pharmaceutical
factors determining absorption rate of a drug
from the liver surface, for the clinical use.
Consequently, drug application to the liver
surface could improve availability in the desired
site of a new drug such as bioactive compound and
genome medicine, by combination with appropriate
chemical and pharmaceutical formulation
modification.
41
Schematic diagram of liver surface application in
rat
Diffusion cell
Area 0.64 cm2 I.D. 9 mm
Model compounds Phenolsulfonphthalein (PSP)
Bromphenol blue (BPB) Bromosulfonphthalein
(BSP) FITC-dextrans (FD-4, FD-10, FD-40, FD-70)
Sampling Plasma Bile Urine Liver Diffusion
cell
Liver
A cylindrical diffusion cell was attached to
the liver surface (left lateral lobe) of male
Wistar rats (250-310 g) with the biocompatible
glue Aron Alpha. Compounds (0.1 ml) were added
into the diffusion cell.
Rat
Nishida et al, J. Pharm. Pharmacol, 46, 867, 1994
42
Plasma concentration and biliary excretion
profiles of PSP, BPB and BSP after application to
rat liver surface
? free PSP ? BPB ? BSP
? PSP metabolite
Biliary excretion rate (µg/min)
Plasma concentration (µg/ml)
Time (min)
Nishida et al, J. Pharm. Pharmacol, 46, 867, 1994
??????
43
Semi-log plot of PSP remaining in diffusion cell
after application to rat liver surface
??????
Diffusion cell
ka 6.92 X 10-3 min-1
ka
(r2 0.99)
kel
Central
Remaining amount ( of dose)
k21
k12
Peripheral
Pharmacokinetic model
Nishida et al, J. Pharm. Pharmacol, 47, 227, 1995
Time (min)
44
Relationship between Mw and ka of model compounds
??????
PSP
BPB
A, B Constant Pa Partition constant
ka (min-1 X 10-3)
BSP
FD-10
FD-4
r2 0.96 Mw 71195 (ka 0)
FD-40
FD-70
Nishida et al, J. Drug Target., 4, 141, 1996
45
First-order absorption rate constant ka of
compounds having different lipophilicity
??????
Log PC
Compounds
(-1.39)
Paraben
Log Ka
(-1.09)
PSP
(-0.60)
Sulfamethizole
(0.19)
Antipyrine
(1.66)
Methylparaben
(2.47)
Propylparaben
(3.41)
Butylparaben
(4.19)
Sudan blue
(4.62)
Sudan III
0
20
40
60
80
100
120
ka ( x 10-3 min-1)
46
Scheme of epithelial cells of liver surface
membrane
A Transcellular route B Paracellular route
J.A. Nagy, Kidney Int. 50 S2 (1996)
47
Liver concentration of PSP at 30 min after
application to liver surface (LSA) or i.v.
administration in rats
Site 1 Site 2 Site 3

Site 2
Site 3
Site 1
Concentration (µg/g liver)
Nishida et al, Pharm. Res., 22, 1331, 2005
Site 1 ????(??????) Site 2 ??????????? Site 3
????

LSA 30 min
i.v. 30 min
p lt 0.01, p lt 0.001, vs. site 3 p lt 0.001,
vs. site 2
FD-70????????????? (?????????2 hr?)
48
Liver concentration and AUC of FD-4 after
application to liver surface or i.v.
administration in rats
Liver concentration
AUC until 360 min
,
Site 1 Site 2 Site 3
p lt 0.05, p lt 0.01, vs. i.v. p lt 0.01, vs.
site 2 p lt 0.01, vs. site 3
Concentration (µg/g liver)
AUC/absorbed amount ( 103 µg/g min mg-1)


LSA 60 min
i.v. 60 min
i.v.
LSA
??????
Nishida et al, Pharm. Res., 22, 1331, 2005
49
Physiological model for PSP distribution after
application to rat liver surface
Concentration (µg/g liver or ml)
Time (min)
??????
50
Absorption of 5-FU from diffusion cell after
application to rat liver surface
Amount in diffusion cell
Mw dependency
5-FU
50
r2 0.95
Papp (µm/min)
Amount ( of dose)
PSP
BPB
Ka 3.3 X 10-3 min-1
FD-4
BSP
FD-10
0.1
0.08
0.06
0
0.02
0.04
FD-70
Time (min)
??????
FD-40
Kodama et al, Biol. Pharm. Bull., 31 1049, 2008
51
Plasma concentration profiles and distribution in
liver of 5-FU after application to rat liver
surface
Plasma profile
Distribution in liver at 60 min
1.2
1.0
?LSA ?i.v.
0.8
Concentration (µg/g liver)
Plasma concentration (µg/ml)
0.6
N.D. not detected
0.4
0.2
N.D.
N.D.
0
Site 1
Site 2
Site 3
Time (min)
??????
Nishida et al, Pharm. Res., 22, 1331, 2005
52
Plasma concentration profiles of PSP after
application to rat liver surface with or without
viscous additives
10
p lt 0.05, p lt 0.01, vs. control
? Control ? CMC-Na 1 ? CMC-Na 3 ? PVA
15 Fitting curve
Plasma concentration (µg/ml)
8


6
Ka (min-1 X 10-3)
4

2
0
Control
PVA 15
Time (min)
CMC-Na 1
CMC-Na 3
??????
Nishida et al, Eur. J. Pharm. Biopharm., 50, 397,
2000
53
Relationship between Mw and Papp of model
compounds after application to several organ
surfaces
Kidney
Small intestine
Liver
r20.979
r20.910
r20.997
Papp (µm/min)
Papp (µm/min)
Papp (µm/min)
Cecum
Stomach
??????
r20.998
r20.999
Papp (µm/min) permeability coefficient Pa
partition coefficient B,C constant about
diffusion
Papp (µm/min)
Papp (µm/min)
Nishida et al, J. Pharm. Pharmacol., 57, 1073,
2005 Eur. J. Pharm. Biopharm., 58, 705, 2004 J.
Pharm. Pharmacol., 54, 1005, 2002 etc
54
Comparison of absorption clearance CLa of model
compounds after application to several organ
surfaces
Stomach
Aorgana) CLa(µl/min)
Route
Liver
(cm2) PSP FD-4 FD-10
Small intestine
186.3 296.2 48.8 22.0 Liver 71.7 77.9 18.4 10.9 K
idney 13.0 32.1 9.2 5.1 Stomach 16.3 20.1 4.0 1.2
Cecum 25.2 28.2 5.3 2.1
Blood
Small intestine
Cecum
Kidney
CLa Papp Aorgan
a) Aorgan from previous papers (Flessner et al).
Peritoneal cavity
Nishida et al, J. Pharm. Pharmacol., 57, 1073,
2005
55
????????????
FD??? ????
FD-4
FD-4??????????????
Control CMC-Na 1 PVA 15
56
????????????
????????????
Kawakami et al, BBRC, 294, 46, 2002
Hirayama et al, Pharm. Res., 20, 328, 2003
???????(???) ??????????????????
57
???????HP http//www.ph.nagasaki-u.ac.jp/
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??????HP http//www.ph.nagasaki-u.ac.jp/lab/dds/in
dex-j.html