Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Reexamination Certificate
1995-10-30
2002-09-24
Wang, Andrew (Department: 1635)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C536S023100, C536S023200, C536S024300, C536S024310, C536S024330, C435S006120
Reexamination Certificate
active
06455689
ABSTRACT:
The present invention is related to antisense-oligonucleotides or effective derivatives thereof hybridizing with an area of a gene coding for transforming growth factor-&bgr; (TGF-&bgr;) , oligonucleotides as nonsense control nucleotides, a pharmaceutical composition comprising at least one anti-sense-oligonucleotide or effective derivatives thereof hybridizing with an area of a gene coding for TGF-&bgr; as well as a use of antisense-oligonucleotides for the manufacturing of a pharmaceutical composition for the treatment of tumors and/or the treatment of the immunosuppressive effect of TGF-&bgr;.
The transforming growth factor-&bgr; (TGF-&bgr;) is a factor which is, for example, secreted by human glioma cells. Human gliomas such as glioblastoma are human tumors for which at present no satisfactory therapy exists. The TGF-&bgr; supports in an autocrine manner the growing of the respective tumor cells. The factor shows immunosuppressive effects and reduces (the proliferation of such cytotoxic T-lymphocytes which otherwise would be able to destroy the glioma cells.
The supression of immune responsiveness has been well documented in patients with malignant gliomas. These patients express a variety of immunological deficiencies including cutaneous anergy, depressed antibody production, diminished numbers of circulating T-cells (Brooks, W. H., Netsky, M. G., Horwitz, D. A., Normansell, D. E. Cell mediated immunity in patients with primary brain tumors, J. Exp. Med., 136: 1931-1947, 1972 and Roszman, T., Elliott, L., Brooks, W. Modulation of T-cell function by gliomas, Immunol. Today 12: 370-374, 1991). More recent studies indicate that these impairments may result from malfunctions in physiological pathways required for normal T-cell activation and from quantitative and qualitative defects in T-cell subsets.
In Proceedings of the 82nd Annual meeting of the American Association for Cancer Research, Houston Tex., USA, May 15-18, 1991, Proc AM ASSOC CANCER RES ANNU MEET 32 (O), 1991, 427 is disclosed that factor-&bgr;-antisense-oligonucleotides inhibit a human melanoma cell line under serum-enriched and stimulate under serum-free culture conditions. The results established indicate different roles of cellular TGF-&bgr;
1
in the growth regulation of HTZ-19-cells depending on the amount of serum present in the culture medium. In addition this may indicate the biological potential and possible draw-backs of exogenously administered TGF-&bgr;-antisense.
J. EXP. MED. 174 (4), 1991, 925-930, Hatzfield J. et al, “Release of early human hematopoietic progenitors from quiescene by antisense transforming growth factor &bgr;-1 or Rb oligonucleotides” discloses release of early human hematopietic progenitors from quiescence by antisense transforming growth factor &bgr;1or Rb oligonucleotides. Rb antisense TGF-&bgr; negatively regulates the cycling status of early hematopoietic progenitors through interaction with the Rb gene product.
Proceedings of the National Academy of Sciences of USA, Vo. 88, February 1991, Washington US, pages 1516-1520, Potts, J. et al., “Epithelial-mesenchymal transformation of embryonic cardiac antisense oligodeoxynucleotide to transforming growth factor beta 3′” discloses that epithelial-mesenchymal transformation of embryonic cardiac endothelial cells is inhibited by a modified antisense oligodeoxynucleotide to transforming growth factor &bgr;3. The transformation depends on the activity of a transforming growth factor &bgr; (TGF-&bgr;) molecule produced by the heart. Modified antisense oligodeoxynucleotides generated to non-conserved regions of TGF-&bgr;1, -2, -3 and -4 were prepared in order to examine the possible roles of these members in this transformation. As a result it has been shown that a specific member of the TGF-&bgr; family (TGF-&bgr;3) is essential for the epithelial-mesenchymal transformation.
WO-A 92/17206 discloses a composition for use in the treatment of wounds to inhibit scar tissue formation during healing comprising an effective activity-inhibitor amount of a growth factor neutralising agent or agents specific against only fibrotic growth factors together with a pharmaceutically acceptable carrier. The method of preparation of said composition and method of administering the composition to a host suffering from tissue wounding is also disclosed.
WO-A 90/09180 discloses methods useful in autologous bone marrow transplantation and cancer therapy. Bone marrow cells from a patient having cancer are treated with selected antisense oligonucleotides in order to deplete the bone marrow of malignant cells prior to infusion back into the bone marrow donor.
It is an object of the present invention to provide a method for the treatment of cancer cells which are correlated with an immunosuppression. Another object of the present invention is to provide an effective agent which inhibits the growth of tumor cells which are related to an immunosuppression.
According to the invention antisense-oligonucleotides or effective derivatives thereof which hybridizes with an area of gene region coding for transforming growth factor-&bgr; (TGF-&bgr;) comprising the following nucleic acid sequences identified in the sequence listing under SEQ ID NO. 1-56 and 137 or comprising the following nucleic acid sequences identified in the sequence listing under SEQ ID NO. 57 to 136 each of the nucleic acids having a DNA- or RNA-type structure are able to solve the problems addressed above. Preferably, the antisense-oligonucleotides hybridize with an area of a gene region coding for growth factor-&bgr;
1
, -&bgr;
2
and/or &bgr;
3
. The anti-sense-oligonucleotide is either able to hybridize with areas of a gene region coding for TGF-&bgr; and/or areas of a gene region coding and non coding for TGF-&bgr;. For example, some nucleotides of the antisense-oligonucleotide sequence hybridizing with an area of a gene region coding for transforming growth factor-&bgr; is hybridizing with an area which does not code for the transforming growth factor whereas, the other part of the respective sequence does hybridize with a gene region coding for TGF-&bgr;. Of course, it is also in the scope of the present invention that the antisense-oligo-nucleotide hybridizes with an area of a gene region just coding for growth factor-&bgr;. It is also understood by the skilled person that fragments having subsequences of the antisense-oligonucleotide works according to the invention so long as production of TGF-&bgr; is reduced or inhibited.
In a preferred embodiment of the present invention the antisense-oligonucleotide or effective derivative thereof is a phosphorothioate-oligodeoxynucleotide.
According to the invention the antisense-oligonucleotides are obtainable by solid phase synthesis using phosphite triester chemistry by growing the nucleotide chain in 3′-5′ direction in that the respective nucleotide is coupled to the first nucleotide which is covalently attached to the solid phase comprising the steps of
cleaving 5′DMT protecting group of the previous nucleotide,
adding the respective nucleotide for chain propagation,
modifying the phosphite group subsequently cap unreacted 5′-hydroxyl groups and
cleaving the oligonucleotide from the solid support,
followed by working up the synthesis product.
The chemical structures of oligodeoxy-ribonucleotides are given in
FIG. 1
as well as the respective structures of antisense oligo-ribonucleotides are given in FIG.
2
. The oligonucleotide chain is to be understood as a detail out of a longer nucleotide chain.
In
FIG. 1
lit. B means an organic base such as adenine (A), guanin (G), cytosin (C) and thymin (T) which are coupled via N9(A,G) or N1(D,T) to the desoxyribose. The sequence of the bases is the reverse complement of the genetic target sequence (mRNA-sequence). The modifications used are
1. Oligodeoxy-ribonucleotides where all R
1
are substituted by
1.1 R
1
=O
1.2 R
1
=S
1.3 R
1
=F
1.4 R
1
=CH
3
1.5 R
1
=OEt
2. Oligodeoxy-ribonucleotides where R
1
is varied at the internucleotide p
Bogdahn Ulrich
Brysch Wolfgang
Schlingensiepen Georg-Ferdinand
Schlingensiepen Karl-Hermann
Schlingensiepen Reimar
Biognostik Gesellschaft für Biomolekulare Diagnostik mbH
Jacobson & Holman PLLC
Lacourciere Karen
Wang Andrew
LandOfFree
Antisense-oligonucleotides for transforming growth... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Antisense-oligonucleotides for transforming growth..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Antisense-oligonucleotides for transforming growth... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2878640