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Z. Jian et al. / Bioorg. Med. Chem. Lett. 13 (2003) 2531–2534
1
1
1
2. Huang, Z. H.; Chen, S.; Huang, J. L. J. Appl. Polym. Sci.
999, 73, 1379.
3. Chen, S. Studies on Synthesis and Properties of Polymeric
Targeting Antitumor Medicines Containing 5-FU and Nitrogen
Mustard; PhD Dissertation Fudan University, Shanghai,
1
1
998.
4. Yokoyama, M.; Okano, T.; Sakurai, Y.; Kikuchi, A.;
Ohsako, N.; Nagasaki, Y.; Kataoka, K. Bioconjugate Chem.
1
1
2
1
992, 3, 275.
5. Huang, Z. H.; Lin, Z. L.; Huang, J. L. Eur. J. Med. Chem.
001, 36, 863.
6. Selected data for compound 7: 1H NMR (500 MHz,
CDCl ) d 1.87 (m, 2H, CH CH CH ), 2.30–2.40 (4H,
3
2
2
2
CH
(
2
CH
2
CH
2
), 3.40–3.80 (CH
2
2
CH O of PEG backbone), 5.64
2
br s, 2H, NH ), 6.71 (d, 2H, phenyl of chlorambucil), 4.1 (t,
4
1
1
H, CH OCO) 7.07 (d, 2H, phenyl of chlorambucil), 8.47 (q,
2
H, pyrimidyl), 8.81 (d, 2H, pyrimidyl).
7. Selected data for compound 8: 1H NMR (500 MHz,
CDCl
OCCH CH CH ), 2.55 (t, 2H, CH CH CH Ph), 3.38 (s, 3H,
3 2 2 2
) d 1.90 (m, 2H, CH CH CH ), 2.34 (t, 2H,
2
2
2
2
2
2
Figure 2. Curves of drug inhibition against tumor growth. aThe
volume of tumor was determined by the following equation:
volume=1/2ÂlengthÂwidth .
CH
CH
O), 3.50–3.80 (CH
CH
3
2
2
O of PEG backbone), 4.23 (t, 2H,
2
OCO), 6.62 (d, 2H, phenyl), 7.06 (d, 2H, phenyl).
2
1
1
8. Veronese, F. M. Biomaterials 2001, 22, 405.
9. In vitro cytotoxity of these polymer drugs was determined
by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT) method. The polymer drugs were sterilized
and then seven sample solutions with the concentration of 10,
showed higher antitumor activity against C6 human
breast cancer cells, however, no sharp difference was
found comparing with the same polymers but without
the sulfadiazine. In vivo, however, SPEG-CBL exhib-
ited higher antitumor activity for Lewis lung carcinoma
than the corresponding MPEG-CBL, that means
SPEG-CBL with sulfadiazine end group showed the
obviously directing action.
À1 À2 À3 À4 À5
1
, 10 , 10 , 10 , 10 and 10 mg/mL respectively were
prepared. The human breast cancer cells were cultured in
Dulbecco Eagle’s minimum essential medium (DMEM) sup-
plemented with 100 u/mL penicillin, 100 u/mL streptomycin
ꢀ
and 10% inactivated calf serum at 37 C in a humidified
atmosphere of 5% CO . The cells harvested from log phase
2
5
were digested by 0.25% trypsin and diluted to 1Â10 cells/mL
by DMEM culture solution containing serum. Then the cells
were seeded into a 96-well plate for 200 mL per well. The drug
Acknowledgements
solution was added to each well for 20 mL. The plate was kept
ꢀ
in CO
CO for prescribed time. Then the cultured cells in each well
2
incubator at 37 C in a humidified atmosphere of 5%
We appreciated the financial support from the Pan-Asia
Biotechnology Company.
2
were mixed with 20 mL of MTT solution of 5 mg/mL in
DMEM culture solution without serum and incubated for 4 h
ꢀ
at 37 C in a humidified atmosphere of 5% CO . The top
2
References and Notes
clear solution was got away and 150 mL dimethylsulfoxide
(
DMSO) was added to dissolve the formazan for each well.
1. Ouchi, T.; Hagihara, Y.; Takahashi, K.; Takano, Y.; Igar-
ashi, I. Drug Design Discovery 1992, 9, 93.
After shaken for 10 min by plate shaker, the OD value of each
well was measured on an ELISAspectrophotometer at 490
nm.
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1994, A31, 1019.
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20. The cytotoxic activity of chlorambucil was evaluated by
the same method as that described for the polymer drugs.
21. For the assessment of the acute toxicity, the TA1 mice
were randomly divided into five groups (20/group and female/
male=1:1), polymer drugs and chlorambucil were injected
ipÂ1 into TA1 mice at five different dose levels on day 0. Then
the behavior and death distribution of the test mice were
recorded. The highest death rate appeared on day 1 and the
condition of the survivals was good after 2 weeks. LD50 was
calculated by using Bliss method.
Compat. Polym. 1995, 10, 51.
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6
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22. Lewis lung carcinoma cells were maintained by intraper-
itioneal passage at weekly intervals in C57 mice. In experi-
8
6
ments with tumor cells, 1Â10 cells in 0.1 mL were
transplanted intracutaneously into the right forelimb of C57
mice (10 mice per group) on day 0. The polymer drugs were
sterilized and dissolved in normal saline. The mice bearing the
Lewis lung carcinoma cells were treated by tail vein injection
with a consecutive (qdÂ5) schedule at various doses from day
1. From day 6, the volume of the Lewis lung carcinoma was
measured to evaluate the in vivo antitumor activity of the
polymer drugs.
9
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(