| Project/Area Number |
04454498
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
外科・放射線系歯学
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| Research Institution | Mie University |
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Principal Investigator |
INUI Madoka Mie Univ., Fac.of Medicine, Asso.Prof, 医学部, 助教授 (70159961)
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| Co-Investigator(Kenkyū-buntansha) |
MORI Atushi Mie Univ., Univ.Hospital, Assistant, 医学部・附属病院, 助手 (00239592)
NOMURA Jouji Mie Univ., Univ.Hospital, Lecturer, 医学部・附属病院, 講師 (80172815)
紀平 浩之 三重大学, 医学部附属病院, 助手 (70177905)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1993: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1992: ¥4,900,000 (Direct Cost: ¥4,900,000)
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| Keywords | Malignant Melanoma Cell / Liposome / Cisplatin / Monoclonal Antibody / Fab' fraction / Atomic Absorption Spectrophotometer / モノクロナール抗体 |
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| Research Abstract |
A purified monoclonal antibody (IA6-C2) to a human melanoma cell line from gingiva (HMG) and its Fab' fraction were coupled on Cisplatinum (CDDP) entrapped liposome. The effect of liposomes (Lip-CDDP, Ab-Lip-CDDP, Fab'-Lip-CDDP) against HMG cell was investigated. The Pharmocokinetics of these liposomes in HMG tumor xenografted nude mice were also examined.
A MoAb was purified with protein A agarose affinity chromatography. N-hydroxysuccinimidyl-3-(pridylditio)-propionate (SPDP) was used for conjugation of MoAb to the liposome, and SH radical of MoAb was used for conjugation of Fab' to the liposome. The liposome was produced, mixing of phosphatidylcholine, cholesterol and 3-(2-pyridyldithio) propionyldipalmitoyl-phosphatidylethanolamine (molar ratio, 26/10/0.4). The encapsulated ratio of CDDP was 3% when platinum concentration was measured with atomic absorption spectrophotometer. The releasing CDDP from Lip-CDDP was tested in 37ﾟC culture medium (RPMI 1640 contained 20% FBS) and in 4ﾟC PBS for stability. The releasing ratio was 56% after 24 hrs and 72% after 48 hrs in culture medium, and 12% after 24 hrs and 14% after 48 hrs in PBS.The Lip-CDDP showed to stabilize in 4ﾟC PBS for 48 hrs.
No effect of PBS encapsulated liposome against HMG cell and normal fibloblast indicates components of liposome to be harmless for cell growth. The anti-HMG cell effects of Ab-Lip-CDDP significantly increased to 2.1-fold of Lip-CDDP and 20-fold of CDDP in vitro. The Fab'-Lip-CDDP effect was, however, almost similar with Lip-CDDP.The tissue platinum concentration of nude mice was increased to 1.4-fold of Lip-CDDP and 2-fold of CDDP in HMG tumor, and decreased to 0.7-fold of Lip-CDDP and 0.5-fold of CDDP in kidney.
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