Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Patent
1997-12-01
2000-03-07
Elliott, George C.
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
536 231, C07H 2104
Patent
active
060342358
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a growth inhibitor for leukemia cells comprising an antisense nucleotide derivative.
BACKGROUND ART
Wilms' tumor is a pediatric kidney tumor that occurs as a result of deactivation of both allele of the Wilms' tumor gene (WT1) located on chromosome 11p13 (Call. K. M., et al., Cell 60; 509, 1990). A non-coding upstream sequence of WT1 (C. E. Campbell, et al., Oncogene 9: 583-595, 1994) and a coding region that includes the intron (D. A. Haber, et al., Proc. Natl. Acad. Sci. USA, 88: 9618-9622, 1991) have previously been reported, and they are expected to be involved in the growth and differentiation of the tumor and so forth (D.A. Haber, et al., Proc. Natl. Acad. Sci. USA, 88; 9618-9622, 1991).
However, it was not known that WT1 is involved in the growth of leukemia cells, and that an antisense oligonucleotide derivative to WT1 suppresses and inhibits growth of leukemia cells.
DISCLOSURE OF THE INVENTION
Thus, the present invention provides a growth inhibitor for leukemia cells comprising an antisense nucleotide derivative to Wilms' tumor gene (WT1).
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a graph showing the inhibitory effects of oligonucleotide on the growth of leukemia cell line K562.
FIG. 2 is a graph showing the relationship between the concentrations of oligonucleotides SE3 and AS3 and the growth of leukemia cell line K562.
FIG. 3 is a graph showing the relationship between the concentrations of oligonucleotides SE4 and AS4 and the growth of leukemia cell line K562.
FIG. 4 is a graph showing the time-based effects of oligonucleotides SE3 and AS3 on the growth of leukemia cell line K562.
FIG. 5 is a graph showing the effects of various oligonucleotides on the growth of leukemia cell line K562.
FIG. 6 is a graph showing the inhibitory effects of various oligonucleotides on the growth of leukemia cell line HEL positive for expression of WT1.
FIG. 7 is a graph showing the inhibitory effects of various oligonucleotides on the growth of leukemia cell line THP-1 positive for expression of WT1.
FIG. 8 is a graph showing the inhibitory effects of various nucleotides on the growth of malignant lymphoma cell line U937 negative for expression of WT1.
FIG. 9 is a graph showing the effects of oligonucleotides SE3 and AS3 on the formation of leukemia cell colonies from bone marrow mononuclear cells derived from leukemia patients.
FIG. 10 is a graph showing the effects of oligonucleotides SE3 and AS3 on the formation of granulocytic macrophage colonies from bone marrow mononuclear cells derived from healthy volunteers.
FIG. 11, panel A is a photograph of the results of electrophoresis indicating a decrease in the level of WT1 protein in calls in the case of adding various WT1 antisense oligonucleotides to a culture of K562 cells.
FIG. 11, panel B indicates a decrease in the level of WT1 protein in cells in the case of adding WT1 antisense oligonucleotides to fresh leukemia cells from a patient with AML.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides A leukemia cell growth inhibitor comprising an antisense oligonucleotide derivative to WT1. The antisense oligonucleotide derivatives used in the present invention is an antisense oligonucleotide derivative to WT1, examples of which include that to the transcription capping site of WT1, gene that to the translation starting region, that to an exon or that to an intron.
For example, a nucleotide sequence of a sense DNA strand of the region containing the transcription capping site of WT1 is represented with SEQ ID NO: 9. In addition, a nucleotide sequence of a sense DNA strand of exons 1 to 10 of the region coding for WT1 is represented with SEQ ID NO: 10 to 19. The present invention uses an antisense oligonucleotide derivative to such a nucleotide sequence of the sense DNA strand of WT1. This antisense oligonucleotide derivative is an antisense oligonucleotide derivative comprising 5 to 50 continuous nucleotides and preferably 9 to 30 nucleotides of antisense DNA or RNA chain for WT1,
REFERENCES:
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Inoue Kazushi
Sugiyama Haruo
Yamagami Tamotsu
Elliott George C.
Sugiyama Haruo
Tadamitsu Kishimoto
Wang Andrew
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