Combination of a chemotherapeutic agent and an inhibitor of the TGF-beta system

#This application is the national phase of PCT application PCT/EP2010/061152 having an international filing date of 30 Jul. 2010, which claims benefit of European application No. 09166893.9 filed 30 Jul. 2009. The contents of the above patent applications are incorporated by reference herein in their entirety.

#The entire content of the following electronic submission of the sequence listing via the USPTO EFS-WEB server, as authorized and set forth in MPEP §1730 II.B.2(a)(C), is incorporated herein by reference in its entirety for all purposes. The sequence listing is identified on the electronically filed text file as follows:

#File Name Date of Creation Size (bytes) 313632011600Seqlist.txt Apr. 11, 2011 91,236 bytes

#FIG. 1 presents the reductive effect of a TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30, on the IC50 regarding gemcitabine's cytotoxicity in a dose dependent manner after data normalization. Gemcitabine was added to Hup-T3 cells, a pancreatic carcinoma cell line in concentrations of 5 μM, 2 μM, 800 nM, 320 nM, 128 nM, 51.2 nM, 20.5 nM, 8.2 nM, or 3.3 nM in combination with the TGF-beta 2 antisense oligonucleotide in concentrations of 0 μM (▪), 5 μM (Δ), or 10 μM (∇). 10 μM of the TGF-beta 2 antisense oligonucleotide reduce the IC50 of gemcitabine about 4 to 5× in comparison to 0 μM TGF-beta 2 antisense oligonucleotide.

#FIG. 2 shows the effect of gemcitabine on proliferation and viability, respectively, of Hup-T3 cells and the secretion of TGF-beta 2 after data normalization. Gemcitabine at 5 μM, 500 nM, 50 nM, 5 nM, 0.5 nM, or 0.05 nM has no specific influence, in particular no specific inhibitory and/or stimulatory effect, on TGF-beta 2 secretion. The decrease of TGF-beta 2 secretion (▪) correlates to the proliferation and viability, respectively, of the cells, which decreases at higher gemcitabine concentrations (A) due to the cytotoxic effect of gemcitabine.

#FIG. 3 presents the dose dependent inhibitory effect of TGF-beta 2 antisense oligonucleotide on TGF-beta 2 secretion. TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30, was administered to Hup-T3 cells in concentrations of 0 μM (control), 1 μM, 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM, 60 μM, or 80 μM. No gemcitabine was added to the cells.

#In the present invention, an inhibitor of the TGF-beta system such as an antisense oligonucleotide, in particular a TGF-beta 1 antisense oligonucleotide, TGF-beta 2 antisense oligonucleotide, and/or TGF-beta 3 antisense oligonucleotide, increases the efficiency of a chemotherapeutic agent in a cell, a tissue, and/or an organ of a subject. In a preferred embodiment, such antisense oligonucleotide is part of a pharmaceutical composition together with a chemotherapeutic agent, wherein the chemotherpeutic is for example gemcitabine, 5-fluorouracil, temozolomide, dacarbacine, docetaxel, cisplatin, oxaliplatin, tamoxifen, or irinotecan.

#The pharmaceutical composition of the present invention is applicable for treating a neoplastic disease in any mammal. Examples of mammal include laboratory animals, including rodents such as mice, rats and guinea pigs; farm animals such as cows, sheep, pigs and oats; pet animals such as dogs and cats; and primates such as monkeys, apes and humans. The pharmaceutical composition is most preferably applied in human clinical situations, particularly for treating neoplastic diseases.

#In one embodiment of this invention one or more chemotherapeutic agents and one or more antisense oligonucleotides and/or one or more inhibitors of the TGF-beta system inhibiting the cell proliferation, form a mixture comprising at least two of these components, wherein the components are either in a pure form or together with a pharmaceutically acceptable carrier, filler, lubricant, diluent, excipient, disintegrate, and/or adjuvant.