WO2011012713A1 - Combination of a chemotherapeutic agent and an inhibitor of the tgf-beta system - Google Patents

Combination of a chemotherapeutic agent and an inhibitor of the tgf-beta system Download PDF

Info

Publication number
WO2011012713A1
WO2011012713A1 PCT/EP2010/061152 EP2010061152W WO2011012713A1 WO 2011012713 A1 WO2011012713 A1 WO 2011012713A1 EP 2010061152 W EP2010061152 W EP 2010061152W WO 2011012713 A1 WO2011012713 A1 WO 2011012713A1
Authority
WO
WIPO (PCT)
Prior art keywords
tgf
beta
carcinoma
cancer
antisense oligonucleotide
Prior art date
Application number
PCT/EP2010/061152
Other languages
English (en)
French (fr)
Inventor
Karl-Hermann Schlingenslepen
Frank Jaschinski
Tanja Rothammer
Anneliese Schneider
Original Assignee
Antisense Pharma Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=41278403&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=WO2011012713(A1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Priority to EP10737580.0A priority Critical patent/EP2340308B1/en
Priority to US13/059,623 priority patent/US8476246B2/en
Priority to RU2012107535/15A priority patent/RU2012107535A/ru
Priority to MX2012001244A priority patent/MX2012001244A/es
Priority to CA2769618A priority patent/CA2769618A1/en
Application filed by Antisense Pharma Gmbh filed Critical Antisense Pharma Gmbh
Priority to AU2010277554A priority patent/AU2010277554B2/en
Priority to BR112012002102A priority patent/BR112012002102A2/pt
Priority to CN2010800436805A priority patent/CN102712924A/zh
Priority to JP2012522185A priority patent/JP2013500313A/ja
Priority to IN969DEN2012 priority patent/IN2012DN00969A/en
Publication of WO2011012713A1 publication Critical patent/WO2011012713A1/en
Priority to IL217821A priority patent/IL217821A0/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1136—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against growth factors, growth regulators, cytokines, lymphokines or hormones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/11—Antisense
    • C12N2310/111—Antisense spanning the whole gene, or a large part of it
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2320/00—Applications; Uses
    • C12N2320/30—Special therapeutic applications
    • C12N2320/31—Combination therapy

Definitions

  • This invention refers to a pharmaceutical composition
  • a pharmaceutical composition comprising a chemotherapeutic agent and an antisense oligonucleotide, preferably a TGF-beta antisense oligonucleotide, or any inhibitor of the TGF-beta system, wherein the antisense oligonucleotide or the inhibitor reduces the IC 5 O of the chemotherapeutic agent ' s cytotoxicity, and thus, increases the efficiency of the chemotherapeutic agent.
  • the invention is further directed to the use of the pharmaceutical composition for the preparation of a medicament for treating a neoplastic disease such as cancer, preferably pancreatic cancer, bladder cancer, glioma, astrocytoma, melanoma, renal carcinoma, lung cancer, breast cancer, ovary cancer, prostate cancer, colorectal cancer, gastric cancer, endometrial cancer, and osteosarcoma as well as to methods for production of such pharmaceutical composition.
  • a neoplastic disease such as cancer, preferably pancreatic cancer, bladder cancer, glioma, astrocytoma, melanoma, renal carcinoma, lung cancer, breast cancer, ovary cancer, prostate cancer, colorectal cancer, gastric cancer, endometrial cancer, and osteosarcoma as well as to methods for production of such pharmaceutical composition.
  • a neoplastic disease such as cancer, preferably pancreatic cancer, bladder cancer, glioma, astrocytoma, melanoma, renal carcinoma, lung
  • chemotherapeutic agent for the preparation of a medicament or radiation is the most common means, beside surgery, for the treatment of neoplastic diseases.
  • a chemotherapeutic agent is for example an alkylating agent, an antimetabolite or an alkaloid derived from a plant.
  • the effect of these chemotherapeutic agents and radiation is the unspecific inhibition of the cell proliferation and the unspecific induction of cell death, respectively, leading to numerous severe side effects.
  • a chemotherapeutic agent or radiation inhibits for example proliferation of rapidly growing cells, other than tumor cells, such as hair follicle, colon mucosa cells or immune cells, e.g., T-lymphocytes, B-lymphocytes, natural killer cells, granulocytes, macrophages, microglia cells as well as the respective precursor cells of the bone marrow.
  • T-lymphocytes e.g., T-lymphocytes, B-lymphocytes, natural killer cells, granulocytes, macrophages, microglia cells as well as the respective precursor cells of the bone marrow.
  • chemotherapeutic agents for the preparation of a medicament for treating cancer and/or the use of radiation do not lead to a sufficient result in the prolongation of the survival of a patient, in particular the median survival, which is sometimes enforced by the severe side effects of the chemotherapeutic agent and/or radiation.
  • WO 2005/059133 A2 refers to pharmaceutical compositions comprising an antineoplastic chemotherapeutic agent and a stimulator of the immune system, which led to an increased cytocytoxicity of lymphokine activated killer cells (LAK cells) on glioma cells in comparison to a stimulator of the immune system alone.
  • LAK cells lymphokine activated killer cells
  • Paz-Ares et al., 2006 disclose the use of a combination of gemcitabine, cisplatin, and a protein kinase C-alpha antisense oligonucleotide for the preparation of a medicament for treating non-small-cell lung cancer.
  • the use of the combination of these chemotherapeutic agents with the antisense oligonucleotide did not enhance the survival or show any other positive effect for the patient suffering from non-small-cell lung cancer.
  • WO 02/17852 A2 describes the combination of a bcl-2 antisense oligonucleotide with a chemoagent in specific administration doses, wherein the bcl-2 antisense oligonucleotide is administered to the patient at high doses for a short period of time, i.e., 14 days.
  • Bcl-2 is an inhibitor of apoptosis since the chemotherapeutic agents described are inducers of apoptosis. Tumor cells become resistant to the chemotherapeutic agents by upregulating bcl-2.
  • the objective problem underlying the present invention is improvement of efficiency of chemotherapeutic agents and pharmaceutical compositions comprising such chemotherapeutic agent, respectively, in the treatment of neoplastic diseases, in particular in non-resistant cells.
  • the present invention refers in one embodiment to a pharmaceutical composition
  • a pharmaceutical composition comprising a chemotherapeutic agent and an inhibitor of the TGF-beta system, preferably an antisense oligonucleotide, e.g., an antitumoral antisense oligonucleotide, wherein the antisense oligonucleotide surprisingly leads to a reduction of the IC 50 of the cytotoxicity of the chemotherapeutic agent in a dose-dependent manner on the chemotherapeutic agent ' s cytotoxicity.
  • an antisense oligonucleotide e.g., an antitumoral antisense oligonucleotide
  • a preferred antisense oligonucleotide is a TGF-beta antisense oligonucleotide like a TGF-beta 1, TGF-beta 2, or TGF-beta 3 antisense oligonucleotide, or combinations thereof, which hybridise with an area of the messenger RNA (m-RNA) and/or DNA encoding TGF-beta 1, -2 and/or -3, or hybridise with m-RNA and/or DNA encoding a TGF-beta 1, -2, and/or -3 receptor.
  • m-RNA messenger RNA
  • the inhibitor of the TGF-beta system is for example selected from the group consisting of TGF-beta binding proteins that are no antibodies, TGF-beta binding receptors, parts of TGF-beta binding receptors, TGF-beta specific peptides and low molecular substances binding TGF-beta or any of their proteins, receptors, part of receptor protein or low molecular substance inhibiting the function of TGF-beta.
  • the inhibitor of the TGF-beta system e.g., the antisense oligonucleotide results in a reduction of the IC 50 of the chemotherapeutic agent ' s cytotoxicity in a resistant or a non- resistant cell, i.e., a cell, which is resistant or non-resistant to the chemotherapeutic agent.
  • the pharmaceutical composition of this invention is in particular for treating neoplastic diseases like cancer and for the treatment of autoimmune diseases.
  • the pharmaceutical composition is used for the preparation of a medicament for treating a neoplastic disease, preferably cancer.
  • the pharmaceutical composition and/or its compounds are prepared in any dosage form and are administered in any route of administration known in the art.
  • the pharmaceutical composition is suitable as a first line treatment of a neoplastic disease like cancer, or as a second, third etc. line treatment before, after or in combination with therapeutic treatments such as radiation.
  • the chemotherapeutic agent and the inhibitor of the TGF-beta system of the pharmaceutical composition are administered either separately or together in one formulation. If more than one chemotherapeutic agent and/or more than one inhibitor of the TGF-beta system, e.g., an antisense oligonucleotide is administered, the chemotherapeutic agent and/or inhibitors of the TGF-beta system such as an antisense oligonucleotide are administered separately or together in one formulation.
  • the administration of the inhibitor of the TGF-beta system such as an antisense oligonucleotide, follows or preceeds the administration of the chemotherapeutic agent, or the inhibitor of the TGF-beta system and the chemotherapeutic are administered concurrently.
  • the effectivity of the chemotherapeutic agent is increased, and in a preferred embodiment the amount and dose, respectively, of the chemotherapeutic agent is reduced resulting advantageously in reduced severe negative side effects of the chemotherapeutic agent.
  • the combination of the chemotherapeutic agent and the inhibitor of the TGF-beta system, for example the antisense oligonucleotide, in the present pharmaceutical composition leads to an advantageous extension of the patient ' s life time based on the supraadditive and synergistic, respectively, antitumoral effect of the compounds.
  • the chemotherapeutic agent does not negatively effect the interaction of the inhibitor of the TGF-beta system, e.g., an antisense oligonucleotide with its target.
  • the present invention further relates to methods for the production of the pharmaceutical composition.
  • Figure 1 presents the reductive effect of a TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30, on the IC 50 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 ⁇ M, 2 ⁇ M, 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 ⁇ M ( ⁇ ), 5 ⁇ M ( ⁇ ), or 10 ⁇ M (V).
  • TGF- beta 2 antisense oligonucleotide reduces the IC 50 of gemcitabine about 4 to 5 x in comparison to 0 ⁇ M TGF-beta 2 antisense oligonucleotide.
  • Figure 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 ⁇ M, 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 ( ⁇ ) 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 ⁇ M (control), 1 ⁇ M, 2.5 ⁇ M, 5 ⁇ M, 10 ⁇ M, 20 ⁇ M, 40 ⁇ M, 60 ⁇ M, or 80 ⁇ M. No gemcitabine was added to the cells.
  • Figure 4 demonstrates that gemcitabine does not affect the interaction of the TGF-beta 2 antisense oligonucleotide with its target, and thus, does not affect the inhibitory effect of the antisense oligonucleotide on its target.
  • the antisense oligonucleotide inhibited TGF-beta 2 secretion in a dose dependent manner (0 ⁇ M ( ⁇ ), 5 ⁇ M ( ⁇ ), or 10 ⁇ M (•) TGF-beta 2 antisense oligonucleotide) in the presence of gemcitabine (2 ⁇ M, 800 nM, 320 nM, 128 nM, 51.2 nM, 20.5 nM, 8.2 nM).
  • FIG. 5 shows the reductive effect of a TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30, on the IC 50 regarding temozolomide ' s cytotoxicity in a dose dependent manner after data normalization.
  • Temozolomide was added to MEL-Juso cells, a melanoma cell line, in concentrations of 200 ⁇ M, 100 ⁇ M, 50 ⁇ M, 25 ⁇ M, 12.5 ⁇ M, 6.25 ⁇ M, 3.125 ⁇ M, or 0 ⁇ M in combination with the TGF-beta 2 antisense oligonucleotide in concentrations of 0 ⁇ M ( ⁇ ), 5 ⁇ M (A), or 10 ⁇ M (T).
  • Figure 6 demonstrates the effect of a TGF-beta 2 antisense oligonucleotide on the cytotoxicity of temozolomide.
  • Temozolomide was administered to A-172 cells, a glioma cell line, in concentrations of 200 ⁇ M and 800 ⁇ M, respectively, either alone or in combination with 10 ⁇ M of a TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30.
  • the combination of temozolomide with the TGF-beta 2 antisense oligonucleotide increased the cytotoxic effect of temozolomide significalty, about 2 to 3 x in comparison to temozolomide alone.
  • 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.
  • 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.
  • a chemotherapeutic agent 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.
  • 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.
  • 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.
  • the one or more chemotherapeutic agents and the one or more inhibitors such as an antisense oligonucleotide are separate in one
  • the pharmaceutical composition comprises each of these components in a pure form or together with a pharmaceutically acceptable carrier, filler, lubricant, diluent, excipient, disintegrate, and/or adjuvant, wherein the pharmaceutically acceptable carrier, lubricant, diluent, excipient, disintegrate and/or adjuvant.
  • the antisense oligonucleotide is preferably any TGF-beta antisense oligonucleotide, which reduces the IC 50 Of the cytotoxicity of the chemotherapeutic agent, and thus, increases the sensitivity of the cell, tissue, and/or organ to the chemotherapeutic agent in vitro, ex vivo, or in vivo.
  • Such antisense oligonucleotides are for example directed against prostaglandine E2 (PGE, e.g., SEQ ID NO. 79-89), VEGF (e.g., SEQ ID NO.
  • TGF-beta 1 e.g., SEQ ID NO. 1-21
  • TGF-beta 2 e.g., SEQ ID NO. 22-48
  • TGF-beta 3 e.g., SEQ ID NO. 49-78.
  • the antitumoral antisense oligonucleotide is an oligonucleotide affecting a tumor, wherein the antitumoral antisense oligonucleotide affects a tumor directly or indirectly.
  • the antisense oligonucleotide blocks the transcription and expression, respectively, of a protein or peptide, which is for example a biological factor in the tumor, e.g., the production of TGF-beta, in particular of TGF beta 2.
  • the antisense oligonucleotide affects the transcription and/or expression of a protein or peptide, for example a factor inducing the function of an immune cell and/or the immune system and in consequence reducing or inhibiting the tumor cell growth and/or inducing cell death of a cancer cell.
  • the inhibitor of the TGF-beta system such as an antisense oligonucleotide, in particluar the antitumoral antisense oligonucleotide, influences the signal transduction, i.e., leads to an increase or decrease of the signal transduction, of factors involved in tumor formation and/or persistance such as capillary formation in the tumor.
  • Immune cells are for example lymphoid cells, such as T cells, B cells, NK cells (natural killer cells), NK T cells (natural killer T cells), granulocytes, such as neutrophils, eosinophils, basophils, and mononuclear cells such as monocytes, macrophages, dendritic cells and mast cells.
  • lymphoid cells such as T cells, B cells, NK cells (natural killer cells), NK T cells (natural killer T cells), granulocytes, such as neutrophils, eosinophils, basophils, and mononuclear cells such as monocytes, macrophages, dendritic cells and mast cells.
  • a TGF-beta inhibitor is any substance, e.g., a protein, peptide, small molelcule, inhibiting the function of TGF-beta in that any effect that is induced by TG F- beta is inhibited.
  • a TGF-beta inhibitor is a substance inhibiting the production of TGF-beta, a substance binding TGF-beta and/or a substance inhibiting the function of TGF- beta downstream its activation cascade.
  • TGF-beta antagonists see also Wojtowicz-Praga (2003).
  • an inhibitor of the TGF-beta system is any substance able to inhibit the expression or function of TGF-beta, in particular TGF-beta 1, -2, and/or -3.
  • the inhibitor is for example selected from the group consisting of TGF-beta binding proteins that are no antibodies, TGF-beta antibodies, TGF-beta binding receptors, parts of TGF-beta binding receptors, TGF-beta specific peptides and low molecular substances binding TGF-beta or any of their proteins, receptors, part of receptor protein or low molecular substance inhibiting the expression and/or the function of TGF-beta.
  • an inhibitor of the TGF-beta system has a molecular weight of less than about 10 kDa and more than about 1 Da of organic or inorganic origin inhibiting the TGF-beta system.
  • the substance inhibiting the production of TGF-beta is a peptide, a peptide of less than 100 kDa, peptides being part of TGF-beta, a protein, a protein that is not an antibody, and/or a small molecule, e.g. tranilast (N-[3,4-dimethoxycinnamoyl]- anthranilic acid) (Wilkenson, K. A. 2000).
  • tranilast N-[3,4-dimethoxycinnamoyl]- anthranilic acid
  • the peptides being part of TGF-beta are sequences of those given in example 9.
  • Example 9 presents the amino acid sequences of TGF-beta 1, TGF-beta 2 and TGF-beta 3 also published in Mittl (1996).
  • peptides comprise the 112 amino acids counted from the end of the TGF-beta 1, TGF-beta 2 or TGF-beta 3 peptide as described in example 9.
  • the start of those peptides is after the RXXR motif, ending 113 amino acids before the end of the TGF- beta 1, TGF-beta 2 or TGF-beta 3 peptide, in which R is the amino acid Arginin and XX represents any amino acid or is even no amino acid.
  • peptides being part of TGF-beta are parts of the sequences presented in example 9 comprising one to all amino acids of this peptide, in other embodiments preferred peptides comprise about 1-100 amino acids, about 2-50 amino acids, about 3-30 amino acids or about 5-20 amino acids of those peptides.
  • preferred amino acids are those presented in example 7 for TGF- beta 1, TGF-beta 2 and TGF-beta 3 with the respective numbers 1-78.
  • amino acids which are described above comprising or consisting of about 1-50 amino acids, about 1-40, about 2-30, about 3-25, about 4-18, about 5-15 or about 6-12 amino acids.
  • At least one of the basic amino acid selected from the group of Histidin (H), Lysin (K) and/or Arginin (R) is substituted by another basic amino acid selected from this group without loosing its TGF-beta antagonizing effects.
  • At least one of the acid amino acid selected from the group of glutaminic acid (E) and/or asparaginic acid (D) is substituted by its counterpart of this group without loosing its TGF-beta antagonizing effects.
  • TGF-beta The peptides that are part of TGF-beta wherein some amino acids are replaced conservatively compared to their sequences presented in example 9 are also referred to as analogs of TGF-beta 1, TGf-beta 2 and/or TGF-beta 3.
  • TGF-beta 2 and TGF-beta 3 about 1 % to about 30 %, about 2% to about 20%, about 3 % to about 15%, 4 % to about 12 % or about 5 % to about 10 % of the amino acids are replaced conservatively.
  • Amino acid replaced conservatively also referred to as conservative analogs or active derivatives of peptides in the context of this invention means replacing at least one amino acid of a peptide or protein.
  • At least one acid amino acid is replaced by the respective other acid amino acid
  • at least one basic amino acid is replaced by another basic amino acid
  • at least one amino acid with a polar group is replaced by another amino acid with a polar group
  • amino acids with pure carbon side chains are replaced by another amino acid with pure carbone side chain.
  • Peptides and/or proteins conservatively replaced with amino acids are still in the scope of this invention.
  • the peptides described above are single and not in the combination with a chemotherapeutic agent. In yet another embodiment these peptides are used for preparing a pharmaceutical composition with a pharmaceutically acceptable carrier. In yet another embodiment these peptides are comprised by a pharmaceutical composition for the treatment of neoplastic diseases and in yet another embodiment these peptides are used for a method treating neoplastic diseases or used for the preparation of a medicament for treating a neoplastic disease according to this invention.
  • the neoplastic disease is in particular a cancer or a tumor such as pancreatic cancer, bladder cancer, brain tumor, melanoma, renal carcinoma, lung cancer, breast cancer, ovary cancer, prostate cancer, colorectal cancer, gastric cancer, endometrial cancer, osteosarcoma, Myosarcoma, blood born tumors, leukemias, tumor metastasis, hemangiomas, acoustic neuromas, neurofibromas, trachomas, pyogenic, granulomas, psoriasis, astracytoma, acoustic neuroma, blastoma, Ewing ' s tumor, craniopharyngioma, ependymoma, medulloblastoma, glioma, hemangloblastoma, Hodgkins-lymphoma, medullablastoma, leukaemia, mesothelioma, neuroblastoma, neurofibroma
  • the brain tumor is in particular an oligodendroglioma, an anaplastic oligoastrozytoma, a glioblastoma, a brain metastasis, a myeloma, a plasmocytoma, a glioma, or an astrocytoma.
  • TGF-beta inhibitors are receptors and/or parts of it, or an antibody and/or parts of it binding TGF-beta, or a protein and/or peptide binding to TGF- beta and in that way inhibiting the function of TGF-beta.
  • the antibodies are for example commercially available, see e.g. R & D Systems, Inc. The production of those antibodies is well known in the art. Animals such as e.g. chicken, mice, rabbits, goats, are immunized with purified human TGF-beta, and the animals produce an antibody against TGF-beta.
  • the antibodies e.g., IgY
  • affinity chromatography as described for example by Cooper, H. M.
  • TGF-beta antibodies are humanized antibodies as described for example by Carrington (1998).
  • Preferred embodiments of the peptides are e.g. Latency- assosciated peptides, which inhibit one or more or all three isoforms of TGF-beta (TGF-beta 1, TGF-beta 2 and TGF-beta 3).
  • the TGF-beta inhibitor is a protein, peptide or a small molecule inhibiting the function of the TGF-beta receptor, acting extracellularly or intracellular ⁇ .
  • Peptides and proteins are for example produced according to classical methods of peptide and protein synthesis such as Merrifield synthesis or Fmoc synthesis.
  • an antisense oligonucleotide preferably an antitumoral antisense oligonucleotide hybridizes with its target messenger RNA (mRNA) and/or DNA.
  • mRNA messenger RNA
  • the target mRNA and/or DNA is any mRNA and/or DNA that is directly or indirectly involved in the formation of a neoplastic disease such as cancer.
  • the antisense oligonucleotide is a TGF-beta antisense oligonucleotide, for example a TGF-beta 1, TGF-beta 2, and/or TGF-beta 3, or its derivative, which hybridises with an area of the mRNA of TGF-beta and/or the DNA encoding TGF-beta, and thus, inhibit the production of TGF- beta.
  • TGF-beta antisense oligonucleotide for example a TGF-beta 1, TGF-beta 2, and/or TGF-beta 3, or its derivative, which hybridises with an area of the mRNA of TGF-beta and/or the DNA encoding TGF-beta, and thus, inhibit the production of TGF- beta.
  • nucleic acid and “oligonucleotide” refer to multiple nucleotides (i.e. molecules comprising a sugar, e.g. ribose or deoxyribose) linked to a phosphate group and to a variable organic base, which is either a substituted pyrimidine, e.g. cytosine (C), thymine (T) or uracil (U) or a substituted purine, e.g. adenine (A) or guanine (G) or a modification thereof.
  • C cytosine
  • T thymine
  • U uracil
  • purine e.g. adenine (A) or guanine (G) or a modification thereof.
  • the terms refer to oligoribonucleotides as well as oligodeoxyribonucleotides.
  • the terms shall also include oligonucleosides (i.e., a oligonucleotide without the phosphate) and any other organic base-containing polymer.
  • the nucleic acids are double-stranded or single-stranded. Double-stranded molecules are more stable in vivo, while single-stranded molecules have increased activity.
  • the nucleotides have lengths between about 6 and about 100 nucleotides in yet another embodiment the nucleotides have lengths of about 8 to about 40 nucleotides respectively from about 12 to about 32 nucleotides.
  • linked units of a nucleic acid means two entities are bound to one another by any physicochemical means. Any linkage known to those of ordinary skill in the art, covalent or noncovalent, is embraced. Natural linkages, which are those ordinarily found in nature connecting the individual units of a nucleic acid, are most common. The individual units of a nucleic acid are linked, however, by synthetic or modified linkages.
  • the respective ends of this linear polymeric structure are further joined to form a circular structure.
  • open linear structures are generally preferred.
  • the phosphate groups are commonly referred to as forming the internucleoside backbone of the oligonucleotide.
  • the normal linkage or backbone of RNA and DNA is a 3 ' to 5 ' phosphodiester linkage.
  • Antisense oligonucleotides or antitumoral antisense oligonucleotides include oligonucleotides having non-naturally occurring portions with similar function. Such modified or substituted oligonucleotides are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target (e.g. protein), altered intracellular localization and increased stability in the presence of nucleases. Modifications of the oligonucleotides as used herein comprise any chemical modifications of the sugar, the base moiety and/or the internucleoside linkage.
  • oligonucleotides or antitumoral antisense oligonucleotides with a covalently modified base and/or sugar include for example oligonucleotides having backbone sugars which are covalently attached to low molecular weight organic groups other than a hydroxyl group at the 3' and/or 2 ' position and other than a phosphate group at the 5' position.
  • modified oligonucleotides include for example a 2'-0-alkylated ribose group.
  • modified oligonucleotides include sugars such as arabinose instead of ribose.
  • the antisense oligonucleotide in particular the antitumoral antisense oligonucleotide, is heterogeneous in the backbone composition comprising or containing any possible combination of polymer units linked together such as peptide-nucleic acids (which have amino acid backbone with nucleic acid bases).
  • the oligonucleotides are homogeneous in the backbone composition.
  • the substituted purines and pyrimidines of the oligonucleotides include standard purines and pyrimidines such as cytosine as well as base analogs such as substituted bases (Wagner et al. 1996).
  • Purines and pyrimidines include, but are not limited to adenine, cytosine, guanine, thymine, 5-methylcytosine, 2-aminopurine, 2-amino-6-chloropurine, 2,6-diaminopurine, hypoxanthine, and other naturally and non-naturally occurring nucleobases, substituted and unsubstituted aromatic moieties.
  • the single nucleotides in each oligonucleotide or polynucleotide polymer contain the same modifications, contain combinations of these modifications, or combine these modifications with phosphodiester linkages.
  • Methods of rendering oligonucleotide or polynucleotide polymers nuclease resistant include, but are not limited to, covalently modifying the purine or pyrimidine bases.
  • bases are methylated, hydroxymethylated, or otherwise substituted (e.g., glycosylated) such that the oligonucleotides or polynucleotides are rendered substantially acid and nuclease resistant.
  • At least one end-block on the oligonucleotide is a biotin, biotin analog, avidin, or avidin analog. These molecules have the ability to block the degradation of the protected oligonucleotide or polynucleotide and provide means for high affinity attachment of the modified oligonucleotides to the solid support.
  • Avidin and biotin derivatives which are for example used to prepare the reagents of this invention include streptavidin, succinylated avidin, monomeric avidin, biocytin (biotin-epsilon-N-lysine), biocytin hydrazide, amine or sulfhydryl derivatives of 2-iminobiotin and biotinyl-epsilon- aminocaproic acid hydrazide.
  • biotin derivatives such as biotin-N- hydroxysuccinimide ester, biotinyl-epsilon-aminocaproic acid-N-hydroxysuccinimide ester, sulfosuccinimidyl 6-(biotin amido)hexanoate, N-hydroxysuccinimideiminobiotin, biotinbromoacetylhydrazide, p-diazobenzoyl biocytin and 3-(N-maleimidopropionyl)biocytin, can also be used as end-blocking groups on the polynucleotides of the present invention.
  • the ring structure of the ribose group of the nucleotides in the modified oligonucleotide has an oxygen in the ring structure substituted with N-H, N-R (with R being an alkyl or aryl substituent), S and/or methylene.
  • the base units are maintained for hybridization with an appropriate nucleic acid target compound.
  • an oligomeric compound an oligonucleotide mimetic that has been shown to have excellent hybridization properties, is for example a peptide nucleic acid (PNA).
  • PNA peptide nucleic acid
  • the sugar-backbone of an oligonucleotide is replaced with an amide containing backbone, in particular an aminoethylglycine backbone.
  • the nucleobases are bound directly or indirectly to aza nitrogen atoms of the amide portion of the backbone.
  • Representative United States patents that teach the preparation of PNA compounds include, but are not limited to, U.S. Pat. Nos. : 5,539,082; 5,714,331; and 5,719,262. Further teaching of PNA compounds can be found in Nielsen et al. (1991).
  • oligonucleotide backbones include, for example, phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphorotriesters, aminoalkylphosphorotriesters, methyl- and other alky-phosphonates including 3 ' -alkylene phophonates and chiral phosphonates, phosphinates, phosphoramidates, including 3 ' -aminophosphoramidate and aminoalkylphosphoramidates, thionophosphoramidates, thionoa Iky I phosphonates, thionoalkylphosphotriesters, and boranophosphates having norm 3 ' -5 ' linkages, 2 ' -5 ' linked analogs of these, and those having inverted polarity wherein the adjacent pairs of nucleoside units are linked 3 ' -5 ' to 5 ' -3 ' or 2 ' -5 ' to 5 ' -2 ' .
  • Various salts
  • At least one nucleotide of an oligonucleotide is modified as described in one of the modifications above.
  • the modifications cover either the oligonucleotide continuously or irregularly.
  • At least two modifications as described above are combined within one oligonucleotide.
  • the 1 to about 12 or 1 to about 8 or 1 to about 4 or 1 to about 2 oligonucleotides and/or nucleotide linkages at the 3 ' and/or 5 ' end of the oligonucleotide are modified as described above.
  • the antisense oligonucleotides of this invention are hybridizing with a target, e.g. TGF-beta or its subtypes TGF-beta 1, TGF-beta 2, TGF-beta 3, or VEGF, IL-IO, or PGE.
  • a target e.g. TGF-beta or its subtypes TGF-beta 1, TGF-beta 2, TGF-beta 3, or VEGF, IL-IO, or PGE.
  • Antisense oligonucleotides of the sequence listing that comprise additional nucleotides for example about 1 to about 1000 nucleotides, from about 1 to about 500, from about 1 to about 100, from about 1 to about 50, from about 1 to about 20, from about 1 to about 10, from about 1 to about 5 or from about 1 to about 2 nucleotides bound to at least one of the 3 ' and/or 5 ' end, in a preferred embodiment on at least one of the 2 ' and/or 5 ' end, are still within the scope of this invention.
  • the antisense oligonucleotides are synthesized de novo using any of a number of procedures well known in the art resulting in "synthetic antisense oligonucleotides". Such procedures are for example, the b-cyanoethyl phosphoramidite method (Beaucage et al. 1981), or the nucleoside H-phosphonate method (Garegg et al. 1986, Froehler et al. 1986, Garegg et al. 1986, Gaffney et al. 1988). These antisense oligonucleotides are performed by a variety of automated oligonucleotide synthesizers available on the market.
  • antisense oligonucleotides are produced in a large scale in plasmids, (see, e.g., Sambrook, et al. 1989) and separated into smaller pieces or administered as a whole.
  • Antisense oligonucleotides are prepared from existing nucleic acid sequences (e.g., genomic or cDNA) using known techniques, such as those employing restriction enzymes, exonucleases or endonucleases. Antisense oligonucleotides prepared in this manner are referred to as isolated nucleic acids.
  • Antisense oligonucleotides having a modified backbone e.g., phosphorothioate bonds
  • oligonucleotides having a modified backbone e.g., phosphorothioate bonds
  • Aryl- and alkyl-phosphonates are made, e.g., as described in U.S. Pat. No. 4,469,863.
  • Alkyiphosphotriesters, in which the charged oxygen moiety is alkylated as described in U.S. Pat. No. 5,023,243 and European Patent No. 092,574, are prepared by automated solid phase synthesis using commercially available reagents.
  • neoplastic disease refers to a proliferative disorder caused or characterized by the proliferation of cells, which have lost susceptibility to normal growth control.
  • cancer includes benigne and maligne tumors and any other proliferative disorders for example the formation of metastasis. Cancers of the same tissue type in general originate from the same tissue, and are for example divided into different subtypes based on their biological characteristics. Four general categories of cancers are carcinoma, sarcoma, leukemia, and lymphoma. Over 200 different types of cancers are known, and every organ or tissue of the body can be affected.
  • astrocytoma WHO I astrocytoma WHO II, astrocytoma WHO III, blastoma, breast cancer, chordoma, craniopharyngioma, endometrial cancer, ependymoma, Ewing ' s tumor, gastric cancer, germinoma, glioma, glioblastoma, hemangioblastoma, hemangioperycatioma, Hodgkins lymphoma, medulloblastoma, leukaemia, mesothelioma, neuroblastoma, non-Hodgkins lymphoma, pinealoma, retinoblastoma, sarcoma (including angiosarcoma, chondrosarcoma, endothelial sarcoma, fibrosarcoma, leiomyosarcoma, liposarcoma, lymphangioandotheliosarcoma, lyphangiosar
  • metastasis refers to the formation of liver, lung, brain, lymphoma node and/or visceral metastasis.
  • metastasis is treatable by use of the pharmaceutical composition of the presentinvention.
  • a chemotherapeutic agent according to the present invention is a substance inhibiting cell proliferation and/or inducing cell death and in a preferred embodiment further inhibits the formation of metastases.
  • the term chemotherapeutic agent comprises, but is not limited to a chemotherapeutic agent, chemotherapeutic agent supplementary potentiating agents and radioactive agents. Examples for this group are given herein.
  • a chemotherapeutic agent is selected from the group of gemcitabine, telozolomid, nitrosoureas, Vinca alkaloids, antagonists of purine and pyrimidines bases, cytostatic antibiotics, camphotecine derivatives, anti-estrogenes, anti-androgens and analogs of gonadotropin releasing hormon.
  • the group of nitrosoureas comprises ACNU, BCNU, CCNU, and/or HCNU.
  • the antineoplastic chemotherapeutic agent agent is selected from the group of nitrosoureas, e.g. ACNU, BCNU, HCNU and/or CCNU, cytotoxic active antibiotics, e.g. doxorubicin, pegylated liposomal doxorubicin (Caelyx ® ), 5- fluorodeoxyuridine, 5-fluorouracil, 5-fluorouridine, gemcitabine, procarbazine, taxol, taxotere, temozolomide, vinblastine, vincristine.
  • ACNU 3-[(-4-Amino-2- methyl-5-pyrimidinyl)methyl]-l-(2-chloroethyl)-l-nitrosourea hydrochloride, CS-439 HCI, Nidran hydrochloride, Nimustine Hydrochloride, NSC-245382.
  • BCNU is Bischloroethylnitrosourea, the chemical name is N, N ' -bis(2-chlorethyl)-N-nitroso-urea, other names are BiCNU, carmustine.
  • CCNU is l-(2-Chloroethy)-3-cyclohexyl-l-nitrosourea.
  • Synonyms are N-(2-chloroethyl)-N ' -cyclohexyl-N-nitroso-urea, Belustine, Cee NU, Chloroethylcyclohexylnitrosourea, ICIG 1109, Lomustine, NSC 79037.
  • One chemical name for temozolomide is 3,4-dihydro-3-methyl-4-oxoimidazo->5,ld ' 1,2,3, 4-tetrazin-8- carboximide.
  • Other names for temozolomide are Temodal, Temodar, methazolastone, CCRG81045, SCH52365, NSC362856, M&B39836.
  • Synonyms for teniposide are 4 ' -Demethylepipodophyllotoxin, 9-(4,6-O-2-thenylidene-b-D- glucopyranoside), Epipodophyllotoxin, EPT, Teniposide VM-26, VM 26, 5,8,8a,9-Tetrahydro- 5-(4-hydroxy-3,5-dimethoxyphenyl)-9- ⁇ [4,6-O-(2-thienylmethylene)-b-D- glucopyranosyl]oxy ⁇ furo[3 ' ,4 ' :6,7]naphtho[2,3-d]-l,3-dioxol-6(5a/-/)-one.
  • Vinca alkaloids comprise vincristine, vinblastine, vindesine and their active derivatives.
  • the antagonist of the purine and pyrimidine bases is selected from the group of 5-fluorouracile, 5-fluorodeoxiuridine, cytarabine and gemcitabine.
  • the chemotherapeutic agent is selected from the group of doxorubicine and liposomal PEGylated doxorubicin
  • the camphthotecine derivative is selected from the group of irinotecane and topotecane
  • the anti estrogenes are selected from the group of tamoxifen, exemestane, anastrozole and fulvestrant
  • the antiandrogens are selected from the group of flutamide and bicalutamide
  • the antprogesterons are selected from the group of mifepriston
  • the analogs of gonadotropin releasing hormon are selected from the group of leuprolide and gosereline.
  • the at least one antisense oligonucleotide preferably an antisense oligonucleotide selected from the group consisting of TGF-beta 1, TGF-beta 2, and TGF-beta 3, and/or inhibitor if the TGF-beta inhibitor is combined with at least one chemotherapeutic selected from the following group: Acivicin; Aclarubicin; Acodazole Hydrochloride; Acronine; Adozelesin; Adriamycin; Aldesleukin; Altretamine; Ambomycin; Ametantrone Acetate; Aminoglutethimide; Amsacrine; Anastrozole; Anthramycin; Asparaginase; Asperlin; Avastin; Azacitidine; Azetepa; Azotomycin; Batimastat; Benzodepa; Bicalutamide; Bisantrene Hydrochloride; Bisnafide Dimesylate; Bizelesin; Bleo
  • chemotherapeutic agents include: 20-epi-l,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara- CDP-DL-PTBA; arginine
  • Chemotherapeutic agent supplementary potentiating agents are for example tricyclic antidepressant drugs (e.g., imipramine, desipramine, amitryptyline, clomipramine, trimipramine, doxepin, nortriptyline, protriptyline, amoxapine and maprotiline); non-tricyclic anti-depressant drugs (e.g., sertraline, trazodone and citalopram); Ca.sup.++ antagonists (e.g., verapamil, nifedipine, nitrendipine and caroverine); Calmodulin inhibitors (e.g., prenylamine, trifluoroperazine and clomipramine); Amphotericin B; Triparanol analogues (e.g., tamoxifen); antiarrhythmic drugs (e.g., quinidine); antihypertensive drugs (e.g., reserpine); Thio
  • Radioactive agents are Fibrinogen I 125; Fludeoxyglucose F 18 ; Fluorodopa F 18; Insulin I 125; Insulin I 131; Iobenguane I 123; Iodipamide Sodium I 131; Iodoantipyrine I 131; Iodocholesterol I 131; Iodohippurate Sodium I 123; Iodohippurate Sodium I 125; Iodohippurate Sodium I 131; Iodopyracet I 125; Iodopyracet I 131; Iofetamine Hydrochloride I 123; Iomethin I 125; Iomethin I 131; Iothalamate Sodium I 125 Iothalamate Sodium I 131; Iotyrosine I 131; Liothyronine I 125; Liothyronine I 131; Merisoprol Acetate Hg 197; Merisoprol Acetate Hg 203; Meris
  • the pharmaceutical composition of the present invention comprises further compounds to decrease side effects of the chemotherapeutic agent or the inhibitor of the TGF-beta system.
  • Radiation is applied in dosages of about 1 Gy to about 100 Gy, more preferred from about 20 to about 80 Gy and most preferred, e.g. for the treatment of astrocytomas, glioblastomas and gliomas from about 40 to about 60 Gy.
  • the dosage in preferred embodiments is fractionated which means that, from about 0.1 to about 10 Gy or from about 1 Gy to about 5 Gy or from about 1 Gy to about 2 Gy are applied in one session which is repeated several times during about 1 to about 20 weeks, about 2 to about 10 weeks or 4 to about 8 weeks.
  • the chemotherapeutic agent and/or the inhibitor of the TGF-beta system e.g., an antisense oligonucleotide is administered before, after or together with the radiation.
  • One cycle of radiation therapy as well as several cycles of radiation are possible, dependent of the reduction of tumor size.
  • the radiation usually is performed with 60 Co. Radiation with neutrons, protons, negative pi- mesones or neutrone capture is applicable as well. It is clear to someone skilled in the art that the dosage is further dependant on the size of the tumor, the build of the patient and the kind of radiation applied. In special embodiments the dosage is about 2 to about 100 fold higher or lower as described above also dependant from the number of fractions the dosage is applied with.
  • the pharmaceutical composition comprising at least one chemotherapeutic agent and at least one inhibitor of the TGF-beta system such as an antisense oligonucleotide is used in combination with other procedures for the treatment of diseases.
  • a tumor may be treated conventionally with surgery and/or radiation and then the composition comprising the chemotherapeutic agent and at least one antisense oligonucleotide and/or at least one inhibitor of the TGF-beta system according to this invention is subsequently administered to the patient to extend the dormancy of micrometastases and to stabilize respectively reduce any residual neoplastic disease, i.e., a tumor.
  • the pharmaceutical composition comprising at least one chemotherapeutic agent and at least one inhibitor of the TGF-beta system, for example an antisesne oligonucleotide is administered to a site likely to harbor a metastatic lesion (that may or may not be clinically discernible at the time).
  • a sustained release formulation implanted specifically at the site (or the tissue) where the metastatic lesion is likely to be is suitable in these latter instances.
  • the embodiments of the pharmaceutical composition comprising at least one chemotherapeutic agent and at least one inhibitor of the TGF-beta system such as an antisense oligonucleotide, for example a TGF-beta 2 antisense oligonucleotide such as SEQ ID No. 30 administered in an effective amount.
  • an antisense oligonucleotide for example a TGF-beta 2 antisense oligonucleotide such as SEQ ID No. 30 administered in an effective amount.
  • the term "effective amount" of a pharmaceutical composition, a chemotherapeutic agent, and an inhibitor of the TGF-beta system, respectively refers to the amount necessary or sufficient to realize a desired biologic effect. This depends amongst others on the mode of delivery (e.g., local or systemic), time period of the dosage, age, weight, general health, sex and diet of the subject receiving the pharmaceutical composition.
  • the effective amount is that amount that reduces the rate or inhibits altogether formation of neoplastis diseases.
  • an effective amount is that amount which decreases or eliminates the neoplastic disease.
  • an effective amount may be that amount which prevents an increase or causes a decrease in new neoplastic diseases.
  • the effective amount varies depending upon whether the composition is used in single or multiple dosages. Dosages given in this writing are for adults. It is quite clear to someone skilled in the art that these dosages have to be adapted if the human being is a child, a person stressed by a further illness or other circumstances.
  • subject doses of the compounds described herein typically range from about 0.1 ⁇ g to about 10 mg per administration, which depending on the application could be given hourly, daily, weekly, or monthly and any other amount of time therebetween.
  • the doses range from about 10 ⁇ g to about 5 mg per administration or from about 100 ⁇ g to about 1 mg, with 1-10 administrations being spaced hours, days or weeks apart.
  • doses may be used in a range even 2 to 100 fold higher or lower than the typical doses described above. These doses are mainly referring to the treatment of adults; in case of the treatment of a child, the doses have to be reduced as known by a skilled person.
  • the effect of a compound is indicated for example by its IC 50 , the half maximal inhibitory concentration, which represents the concentration of an inhibitor that is required for 50 % inhibition of its target, i.e., it measures how much of a particular substance/molecule is needed to inhibit some biological process by 50 %.
  • the IC 50 of the chemotherapeutic agent describes the concentration of the chemotherapeutic agent that results in 50 % cytotoxicity.
  • the IC 50 describes the efficiency of a compound, the lower the IC 50 of a compound, the more effective the compound.
  • the antisense oligonucleotide in particular the antitumoral antisense oligonucleotide such as TGF-beta 1, -2, or -3 leads to a 1.5x, 2x, 2.5x, 5x, 5.5x, 6x, 6.5x, 7x, 7.5x, 8x, 8.5x, 9x, 9.5x, 1Ox, 15x, 2Ox, 25x, 3Ox, 35x, 4Ox, 45x, 5Ox, 55x, 6Ox, 65x, 7Ox, 75x, 8Ox, 85x, 9Ox, 95x, or 99x reduction of the IC 50 of the chemotherapeutic agent, preferably gemcitabine.
  • the IC 50 of the chemotherapeutic agent such as gemcitabine or temozolomide is 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% reduced by the antisense oligonuclotide, e.g., a TGF-beta 1, -2, and/or -3 antisense oligonucleotide, and/or an inhibitor of the TGF-beta system in comparison to gemcitabine without an antisense oligonucleotide, particularly a TGF-beta 1, -2, and/or -3 antisense oligonucleotide.
  • the antisense oligonuclotide e.g., a TGF-beta 1, -2, and/or -3 antisense oligonucleotide
  • the inhibitor of the TGF-beta system reduces the IC 50 of the chemotherapeutic agent in a dose dependent manner.
  • the chemotherapeutic agent does neither influence the expression or activity of the target of the inhibitor of the TGF-beta system such as the antisense oligonucleotide, which lead to the IC 50 reduction of the chemotherapeutic agent, nor the interaction of the the inhibitor of the TGF-beta system, e.g., the antisense oligonucleotide with the target.
  • the chemotherapeutic agent even supports and increases the interaction of the inhibitor of the TGF-beta system, e.g., the antisense oligonucleotide, with the target.
  • a preferred target of the antisense oligonucleotide is TGF-beta 1, -2, and/or -3.
  • the at least one inhibitor of the TGF-beta system in particular the TGF-beta 1, -2, or -3 antisense oligonucleotide is administered in a dose range from about 1 ⁇ g/kg/day to about 100 mg/kg/day or from about 10 ⁇ g/kg/day to about 10 mg/kg/day or from about 100 ⁇ g/kg/day to about 1 mg/kg/day.
  • the pharmaceutical composition is administered with a catheter directly into the tumor.
  • the concentrations of the antinsense oligonucleotides are from about 0.1 ⁇ M/L to about 1 M/L, more preferred from about 1 ⁇ M/L to about 500 ⁇ M/L and even more preferred from about 10 to about 200 ⁇ M/L or from about 50 ⁇ M/L to about 150 ⁇ M/L in a steril aqueous solution.
  • this solution is administered with a flow of about 0.1 ⁇ L/min to about 50 ⁇ L/min or about 2 ⁇ L/min to about 12 ⁇ L/min or about 3 ⁇ L/min to about 10 ⁇ L/min into the tumor.
  • the at least one chemotherapeutic agent is selected from the group of nitrosourea, more preferred BCNU, CCNU and/or ACNU in combination with at least an inhibitor of the TGF-beta system, e.g., an antisense oligonucleotide such as TGF-betal, - beta2- or -beta3 antisense oligonucleotide.
  • an antisense oligonucleotide such as TGF-betal, - beta2- or -beta3 antisense oligonucleotide.
  • the chemotherapeutic agent such as gemcitabine or temozolomid is for example administerd in a dose range from about 1 mg/m 2 to about 1000 mg/m 2 , more preferred in a dose of about 50 mg/m 2 to about 500 mg/m 2 and most preferred in a single dose of about 150 mg/m 2 to 200 mg/m 2 intravenously every 6 weeks. It may be given as a single dose or divided into daily injections such as about 75 mg/m 2 to about 100 mg/m 2 on two successive days.
  • the chemotherapeutic agent is gemcitabine and is administered with at least an inhibitor of the TGF-beta system such as an antisense oligonucleotide, and/or radiation at a dosage of about 10 mg/m 2 to about 10 g/m 2 , more preferred from about 100 mg to about 5g/m 2 and most preferred from about 500 mg/m 2 to about 2000 mg/m 2 .
  • Gemcitabine is preferably administered within about 10 min to about 120 min, more preferred within about 15 min to about 60 min and most preferred within about 20 min to about 40 min. before or after administration of the antisense oligonucleotides.
  • gemcitabine is coadministered with one or more antisense oligonucleotides at the same time, wherein gemcitabine and the antisense oligonucleotide such as SEQ ID No. 30 are administered separately or in combination.
  • a single dose of the chemotherapeutic agent such as gemcitabine is administered repeatedly within about 4 to about 10 days, respectively about 5 to about 8 days and most preferred within about 7 days.
  • About 1 to about 8, more preferred about 2 to about 6 most preferred about 3 to about 4 single doses are administered within about 4 to about 10 days, respectively about 5 to about 8 days and most preferred within about 7 days.. After this a therapy free interval of about 2 to about 60 days, more preferred about 5 to about 30 days and most preferred from about 10 to about 20 days is applied. Several repetitions of these cycles are possible, e.g., 1 to 10, 2 to 10, 3 to 10, 4 to 10, 5 to 10, 6 to 10, 7 to 10, 8 to 10, or 9 or 10.
  • At least one chemotherapeutic agent is temozolomide and is administered with a total dose of about 500 to about 1200 mg/m 2 , over a period from about 2 to about 28 consecutive days, more preferable over a period of from about 4 to about 7 consecutive days, and most preferably over a period of about 5 consecutive days.
  • the total dose is to be about 1000 mg/m 2 administered over a period of about 5 days
  • the daily dose for this period is about 200 mg/m 2 /day.
  • Temozolomide is administered at least once per day. Preferably dosing regimes would be twice per day, three times per day or four times per day. After a period of about 28 to about 42 days, or about 28 to about 35 days, or more preferably 28 days, from the first day of temozolomide administration, another administration cycle may be started.
  • the temozolomide may be administered for a much longer period at reduced dosage.
  • the temozolomide is administered more than once daily for up to six weeks at a daily dosage of about 50 mg/m 2 /day to about 150 mg/m 2 , of about 50 mg/m 2 /day to about 75 mg/m 2 /day, most preferably of about 75 mg/m 2 /day. More preferred these daily doses are split about evenly into two or more doses to be administered two or more times per day.
  • vinblastin is administered at a dosage of about 0.1 mg/m 2 to about 50 mg/m 2 more preferred in a dose of about 1 mg/m 2 to about 10 mg/m 2 and even more preferred at about 4 mg/m 2 to about 8 mg/m 2 .
  • vincristin is administered at a dose of about 0.1 mg/m 2 to 10 mg/m 2 more preferred in a dose of about 0.5 mg/m 2 to about 5 mg/m 2 and more preferred at about 0.8 mg/m 2 to about 2 mg/m 2 about once a week whereas the neurotoxicity is the dosage limiting factor.
  • vincristin sulfate from about 0.1 mg/mL to about 10 mg/mL are administered with single doses of about 0.1 mg/m 2 to about 50 mg/m 2 more preferred in a dose of about 0.5 mg/m 2 to about 10 mg/m 2 and even more preferred from about 1 mg/m 2 to about 5.0 mg/m 2 .
  • a pharmaceutical composition for the treatment of pancreas carcinoma, glioblastoma and/or anaplastic astrocytoma comprises a combination of at least one antisense oligonucleotide, e.g., a TGF-beta 1, -2, and/or -3 antisense oligonucleotide, preferably a TGF-beta antisense oligonucleotide of SEQ ID NO.
  • a chemotherapeutic agent preferably selected from the group consisting of temozolomide, ACNU, BCNU, CCNU, vinblastine, vincristine, vindesine and their active derivatives, 5- fluorouracile, 5-fluorodeoxiuridine, cytarabine, gemicitabine, liposomal pegylated doxorubicine, procarbazine and vincristin.
  • the chemotherapeutic agents procarbazine, CCNU and vincristin are administered together with an antisense oligonucleotide identified in the sequence listing under SEQ ID NO. 1-127 and even more preferred in SEQ ID NO. 22-48, and/or an inhibitor of the TGF-beta system.
  • the dosage in this embodiment is about 40 mg/m 2 to about 80 mg/m 2 of procarbazine p.o. (days about 8 to about 21 from the beginning of administration), about 80 to about 120 mg/m 2 CCNU, p.o. (about day 1 of administration), vincristine from about 1.2 mg/m 2 to about 1.8 mg/m 2 p.o.
  • the antisense oligonucleotide and/or the inhibitor of the TGF-beta system is given before, with or after the administration of the chemotherapeutic agent, i.e., in general the compounds of the pharmaceutical composition of the present invention are administered at the same time, timely overlapping, or timely distinct. In another embodiment this cycle is repeated after about 6 to about 8 weeks once or several times.
  • the at least one antisense oligonucleotide even more preferred an antisense oligonucleotid of TGF-beta 1, -2, or -3, and most preferred, an antisense oligonucleotide identified in the sequence listing under SEQ ID NO. 1-127 and even more preferred the sequences with SEQ ID NO. 22-48 and telozolomide are the parts of the pharmaceutical composition.
  • the dosage of temozolomide for the treatment of a neoplastic disease, more preferred cancer such as pancreatic carcinoma, glioma, glioblastoma and/or anaplastic astrocytoma is from about 120 to about 180 mg/m 2 , p.o. on day 1 to 5 of a cycle.
  • the antisense oligonucleotide is administered from about 1 ⁇ g/kg/day to about 50 mg/kg/day. The cycle is repeated after about 3 to 5 weeks.
  • radiation is further administered according to standard schedules as described above.
  • the radiation is applied together with the administration of the combination as described above.
  • the radiation is applied before or after the administration of the pharmaceutical compositions according to this invention.
  • At least one chemotherapeutic agent inhibiting cell proliferation and/or inducing cell death is selected from the group of cisplatin, carboplatin, cyclophosphamid, docetaxel, PEG-liposomal doxorubicin, etoposid, folinicc acid, 5- fluorouracil, mitoxantrone, paclitaxel, topotecan and/or treosulfan.
  • the chemotherapeutic agents paclitaxel or carboplatin are the at least one part of a pharmaceutical composition according to this invention.
  • Paclitaxel from about 100 mg/m 2 to about 200 mg/m 2 more preferred about 175 mg/m 2 or carboplatin administered i.v. at day 1 of a cycle. This cycle is repeated after about 20 to about 30 days.
  • the at least one chemotherapeutic agent of a pharamaceutical composition according to this invention is gemcitabine.
  • Gemcitabine is administered in dosages of about 800 mg/m 2 to about 1200 mg/m 2 , more preferred about 1000 mg/m 2 i.v. within about 10 min to about 60 min, more preferred within about 12 min to about 20 min. This application is repeated for about 5 to about 10 days.
  • paclitaxel together with carboplatin docetaxel together with carboplatin, carboplatin together with cyclophosphamid, cisplatin together with treosulfan, etoposid, mitoxantron together with folin acid and 5-fluorouracil, topotect, or PEG-liposomal doxorubicin are the at least one chemotherapeutic agent of a pharamaceutical composition according to this invention for the treatment of pancreatic cancer.
  • the antisense oligonucleotide is an oligonucleotide identified in the sequence listing under SEQ ID NO. 1-127 and even more preferred the sequence with SEQ ID NO. 22-48.
  • the antisense oliogonucleotide is administered in a dose of 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 ⁇ M.
  • radiotherapy is applied according to standard schedules as described above.
  • the composition is administered systemically in a preferred embodiment.
  • the pharmaceutical composition of the present invention preferably comprises at least one chemotherapeutic agent such as gemcitabine or temozolomide and at least one inhibitor of the TGF-beta system such as a TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30, forming the components of the pharmaceutical composition.
  • These components are either in pure form mixed together, or together with a pharmaceutically acceptable carrier, lubricant, diluent, excipient, disintegrate, and/or adjuvant mixed together.
  • the components of the pharmaceutical composition are separate, either in pure form, or together with a pharmaceutically acceptable carrier, lubricant, diluent, excipient, disintegrate, and/or adjuvant.
  • the pharmaceutically acceptable carrier, lubricant, diluent, excipient, disintegrate, and/or adjuvant of the components is identical or different.
  • administering the pharmaceutical compositions of the present invention is accomplished by any means known to a person skilled in the art.
  • Routes of administration include but are not limited to oral, intranasal, intratracheal, ocular, pulmonal, vaginal, rectal, parenteral (e.g. intramuscular, intradermal, intravenous, intratumoral or subcutaneous or direct injection), depo injection, implantation, time-release mode, intracranial, intraperitoneal, intravesical, subconjunctival, topical, transdermal, or sublingual.
  • a pharmaceutical composition for the treatment of neoplastic diseases forming a tumor such as cancer
  • the combination of at least one chemotherapeutic agent and the at least one inhibitor of the TGF-beta system such as an antisense oligonucleotide are preferably delivered by means of a biodegradable, polymeric implant or implanted catheter.
  • compositions refers to compositions comprising the components in solid and/or liquid form, wherein the components are in pure form and/or together with a pharmaceutically acceptable carrier, filler, lubricant, diluent, excipient, disintegrate, and/or adjuvant.
  • Pharmaceutical acceptable carrier, filler, lubricant, diluent, excipient, disintegrate, and/or adjuvant according to the present invention is any substance suitable for administration to a subject, which are of organic or inorganic origin, natural or synthetic origin, and with which a component of the pharmaceutical composition is for example combined to facilitate the application, or to increase the efficiency of the component.
  • a carrier, filler, lubricant, diluent, excipient, disintegrate, and/or adjuvant enables the components of the pharmaceutical composition or the pharmaceutical composition to be formulated as tablet, coated tablet, evervescent tablet, granules, lozenge, powder, pill, dragee, (micro)capsule, liquid, gel, syrup, slurry, suspension, emulsion and the like, for oral ingestion by a subject to be treated.
  • the pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, coated onto microscopic gold particles or preparations with protracted release of the components, in whose preparation excipients and additives and/or auxiliaries such as disinteg rants, binders, coating agents, swelling agents, lubricants, flavorings, sweeteners or solubilizers are customarily used as described above.
  • auxiliaries such as disinteg rants, binders, coating agents, swelling agents, lubricants, flavorings, sweeteners or solubilizers are customarily used as described above.
  • pharmaceutical preparations for oral use are obtained as solid excipient, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores.
  • suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP).
  • fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol
  • cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carb
  • disintegrating agents are added, such as the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
  • the oral formulations may also be formulated in saline or buffers for neutralizing internal acid conditions.
  • dragee cores are provided with suitable coatings.
  • suitable coatings may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
  • dyestuffs or pigments are added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
  • pharmaceutical preparations which can be used orally "vegicaps” include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules containes the active ingredient in a mixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers.
  • the active compounds are dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • suitable liquids such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers may be added.
  • microspheres formulated for oral administration are used, wellknown to someone skilled in the art. The formulations for oral administration are in dosages suitable for such administration.
  • compositions take for example the form of tablets or lozenges formulated in conventional manner.
  • the compounds for use according to the present invention may be conveniently delivered in the form of an aerosol spray, from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • a suitable propellant e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
  • suitable pharmaceutical carriers are, for example, aqueous or saline solutions for inhalation, microencapsulated, encochleated, contained in liposomes, nebulized, aerosols.
  • the pharmaceutical acceptable carriers of the compounds include sterile aqueous solutions which may also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutical acceptable carriers, fillers, lubricants, diluents, excipients, disintegrates, and/or adjuvants.
  • compositions or its components are for example together with a pharmaceutical carrier, filler, lubricant, diluent, excipient, disintegrate, and/or adjuvant for parenteral administration by injection (e.g., by bolus injection or continuous infusion).
  • a pharmaceutical carrier for injection
  • the pharmaceutical compositions take such forms amongst others as suspensions, solutions or emulsions in oily or aqueous vehicles, and contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • pharmaceutical carriers, fillers, lubricants, diluents, excipients, disintegrates, and/or adjuvants for parenteral administration include aqueous solutions of the active compounds in water-soluble form.
  • a suspension of one or more components of the pharmaceutcal composition of the invention is prepared as appropriate oily injection suspension.
  • suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes.
  • Aqueous injection suspensions comprise substances, which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran.
  • the suspension may also contain suitable stabilizers or agents, which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
  • the chemotherapeutic agent and/or the inhibitor of the TFG-beta system e.g., an antisense oligonucleotide is in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use or dried onto a sharp object to be scratched into the skin.
  • a suitable vehicle e.g., sterile pyrogen-free water
  • the compounds are formulated in rectal or vaginal compositions such as suppositories or retention enemas or tablets, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
  • the compounds are formulated as a depot preparation.
  • such long acting formulations are formulated with suitable polymeric or hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example as a sparingly soluble salt.
  • delivery systems include time-release, delayed release or sustained release delivery systems. Such systems can avoid repeated administrations of the compounds, increasing convenience to the subject and the physician. Many types of release delivery systems are available and known to those of ordinary skill in the art.
  • the delivery systems include polymer base systems such as poly(lactide- glycolide), copolyoxalates, polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyric acid, and polyanhydrides.
  • polymer base systems such as poly(lactide- glycolide), copolyoxalates, polycaprolactones, polyesteramides, polyorthoesters, polyhydroxybutyric acid, and polyanhydrides.
  • Microcapsules of the foregoing polymers containing drugs are described in, for example, U.S. Pat. No. 5,075,109.
  • the delivery systems include non-polymer systems that are e.g. lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono-, di- and triglycerides; hydrogel release systems; sylastic systems; peptide based systems; wax coatings; compressed tablets using conventional binders and excipients; partially fused implants; and the like.
  • lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono-, di- and triglycerides
  • hydrogel release systems e.g. lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono-, di- and triglycerides
  • hydrogel release systems e.g. lipids including sterols such as cholesterol, cholesterol esters and fatty acids or neutral fats such as mono-, di- and triglycerides
  • sylastic systems such as cholesterol, cholesterol esters and fatty acids
  • the chemotherapeutic agent and/or the antisense oligonucleotide and/or the inhibitor of the TGF-beta system is formulated with GELFOAM®, a commercial product consisting of modified collagen fibers that degrade slowly.
  • the pharmaceutical compositions also comprise for example suitable solid or gel phase carriers, fillers, lubricants, diluents, excipients, disintegrates, and/or adjuvants.
  • chemotherapeutic agent and/or the antisense oligonucleotide and/or the inhibitor of the TGF-beta system is administered neat or in the form of a pharmaceutical acceptable salt.
  • the salts have to be pharmaceutical acceptable, but non-pharmaceutical acceptable salts may conveniently be used to prepare pharmaceutical acceptable salts thereof.
  • Such salts include, but are not limited to, those prepared from the following acids: hydrochloric, hydrobromic, sulphuric, nitric, phosphoric, maleic, acetic, salicylic, p-toluene sulphonic, tartaric, citric, methane sulphonic, formic, malonic, succinic, naphthalene-2- sulphonic, and benzene sulphonic.
  • such salts can be prepared as alkaline metal or alkaline earth salts, such as sodium, potassium or calcium salts of the carboxylic acid group.
  • suitable buffering agents include but are not limited to: acetic acid and a salt (1-2% w/v); citric acid and a salt (1-3% w/v); boric acid and a salt (0.5-2.5% w/v); and phosphoric acid and a salt (0.8-2% w/v).
  • Suitable preservatives include benzalkonium chloride (0.003-0.03% w/v); chlorobutanol (0.3-0.9% w/v); parabens (0.01-0.25% w/v) and thimerosal (0.004-0.02% w/v).
  • the pharmaceutically acceptable carrier for topical administration for the at least two components of a pharmaceutical composition according to this invention include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders.
  • Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.
  • coated condoms, gloves and the like are useful.
  • the pharmaceutical compositions include penetration enhancers in order to enhance the alimentary delivery. Penetration enhancers may be classified as belonging to one of five broad categories, i.e., fatty acids, bile salts, chelating agents, surfactants and non-surfactants (Lee et al.
  • penetration enhancers from one or more of these broad categories are included.
  • Various fatty acids and their derivatives which act as penetration enhancers include, for example, oleic acid, lauric acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, recinleate, monoolein (a.k.a.
  • fatty acids examples include sodium caprate and sodium laurate, used singly or in combination at concentrations of 0.5 to 5%.
  • the physiological roles of bile include the facilitation of dispersion and absorption of lipids and fat-soluble vitamins (Brunton 1996).
  • the term "bile salt” includes any of the naturally occurring components of bile as well as any of their synthetic derivatives.
  • a presently preferred bile salt is chenodeoxycholic acid (CDCA) (Sigma Chemical Company, St. Louis, Mo.), generally used at concentrations of 0.5 to 2%.
  • CDCA chenodeoxycholic acid
  • bile salts are used in combination with fatty acids to make complex formulations.
  • Preferred combinations include CDCA combined with sodium caprate or sodium laurate (generally 0.5 to 5%).
  • chelating agents include, but are not limited to, disodium ethylenediaminetetraacetate (EDTA), citric acid, salicylates (e.g., sodium salicylate, 5-methoxysalicylate and homovanillate), N-acyl derivatives of collagen, laureth-9 and N-amino acyl derivatives of beta-diketones (enamines) (Lee et al. 1991; Muranishi 1990; Buur et al. 1990). Chelating agents have the added advantage of also serving as DNase inhibitors.
  • EDTA disodium ethylenediaminetetraacetate
  • citric acid citric acid
  • salicylates e.g., sodium salicylate, 5-methoxysalicylate and homovanillate
  • N-acyl derivatives of collagen e.g., laureth-9 and N-amino acyl derivatives of beta-diketones (enamines) (Lee et al. 1991; Muranishi 1990; Bu
  • Sufactants include, for example, sodium lauryl sulfate, polyoxyethylene-9-lauryl ether and polyoxyethylene-20- cetyl ether (Lee et al. 1991); and perfluorochemical emulsions, such as FC-43 (Takahashi et al. 1988).
  • Non-surfactants include, for example, unsaturated cyclic ureas, 1-alkyl- and 1- alkenylazacyclo-alkanone derivatives (Lee et al. 1991); and non-steroidal anti-inflammatory agents such as diclofenac sodium, indomethacin and phenylbutazone (Yamashita et al. 1987).
  • the pharmaceutical compositions of the present invention additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels.
  • the compositions may contain additional compatible pharmaceutically active materials such as, e.g., antipruritics, astringents, local anesthetics or anti-inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the composition of present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers.
  • additional compatible pharmaceutically active materials such as, e.g., antipruritics, astringents, local anesthetics or anti-inflammatory agents
  • additional materials useful in physically formulating various dosage forms of the composition of present invention such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers.
  • such materials when added, should not unduly interfere with the biological activities of the components of the compositions of the invention.
  • TGF-beta 2 antisense oligonucleotide reducing the IC 50 of gemcitabine (Fig. 1)
  • Hup- T3 human pancreatic tumor cell line
  • gemcitabine concentrations i.e., 5 ⁇ M, 2 ⁇ M, 800 nM, 320 nM, 128 nM, 51.2 nM, 20.5 nM, 8.2 nM, 3.3 nM and 0 nM gemcitabine, respectively, in combination with 0 ⁇ M ( ⁇ ), 5 ⁇ M ( ⁇ ) or 10 ⁇ M (V) TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30, for 5 h.
  • gemcitabine concentrations i.e., 5 ⁇ M, 2 ⁇ M, 800 nM, 320 nM, 128 nM, 51.2 nM, 20.5 nM, 8.2 nM, 3.3 nM and 0 nM gemcitabine, respectively, in combination with 0 ⁇ M ( ⁇ ), 5 ⁇ M ( ⁇ ) or 10 ⁇ M (V) TGF-beta 2 antisense oligonucleotide, for example S
  • the first treatment solution comprising gemcitabine and the TGF-beta 2 antisense oligonucleotide was removed and replaced by a second treatment solution containing the TGF-beta 2 antisense oligonucleotide, but no gemcitabine.
  • the treatment solution was optionally replaced after 3 days.
  • TGF-beta 2 concentration was analyzed (see Example 4, and Fig. 4).
  • the proliferation/viability of the Hup-T3 cells was analyzed using the EZ4U method according to the manufacturer's instructions (Biozol Diagnostcamaschine GmbH), and the OD was measured after an incubation time of 75 min with EZ4U solution using the plate reader "Fluostar-Optima" (BMG LABTECH GmbH).
  • the results show an unexpected increase in the inhibition of cell proliferation by gemcitabine, when gemcitabine was administered in combination with the TGF-beta 2 antisense oligonucleotide.
  • the TGF-beta 2 antisense oligonucleotide surprisingly reduced the IC 50 of gemcitabine in a dose dependent manner (Fig. 1).
  • Gemcitabine was administered to the Hup-T3 cells in following concentrations: 5 ⁇ M, 0.5 ⁇ M, 50 nM, 5 nM, 0.5 nM, 0.05 nM or 0 nM, and the cells were incubated for 5 h according to Example 1. The treatment solution was optionally replaced after 3 days.
  • TGF-beta 2 secretion ( ⁇ ) was quantified by a standard TGF-beta 2-ELISA Kit (R&D Systems, Minneapolis, USA) according to the manufacturer's instructions.
  • the proliferation/viability of the Hup-T3 cells was analyzed using the EZ4U method according to the manufacturer's instructions (Biozol Diagnostcamaschine GmbH), and the OD was measured after an incubation time of 75 min with EZ4U solution using the plate reader "Fluostar-Optima” (BMG LABTECH GmbH).
  • TGF-beta 2 secretion correlates to the proliferation and viability ( ⁇ ), respectively, of the cells, which decreases at higher gemcitabine concentrations due to the cytotoxic effect of gemcitabine. This is shown by the overlapping curves of Fig. 2.
  • gemcitabine has a cytotoxic effect, but does not specifically influence the TGF- beta 2 secretion of Hup-T3 cells ( ⁇ ) (see Fig. 2).
  • TGF-beta 2 antisense oligonucleotide for example SEQ ID No. 30, was administered to the Hup-T3 cells as described in Example 1 (0 ⁇ M, 1 ⁇ M, 2.5 ⁇ M, 5 ⁇ M, 10 ⁇ M, 20 ⁇ M, 40 ⁇ M, 60 ⁇ M, or 80 ⁇ M TGF-beta 2 antisense oligonucleotide).
  • the cells were incubated according to Example 1 and after 7 days of treatment, the cell supernatants were removed for the analysis of TGF-beta 2 concentration using the TGF-beta 2-ELISA Kit of R&D Systems.
  • the TGF-beta 2 antisense oligonucleotide inhibited the TGF-beta 2 expression and secretion, respectively, in a dose dependent manner (see Fig. 3).
  • Hup-T3 cells were incubated with different concentrations of gemcitabine (2 ⁇ M, 800 nM, 320 nM, 128 nM, 51.2 nM, 20.5 nM, 8.2 nM, and 0 nM) and TGF-beta 2 antisense oligonucleotide according to Example 1 (0 ⁇ M ( ⁇ ), 5 ⁇ M ( ⁇ ), or 10 ⁇ M (•) TGF-beta 2 antisense oligonucleotide) and the cell supernatants were removed after 7 days of treatment for the analysis of TGF-beta 2 concentration using the TGF-beta 2-ELISA Kit of R&D Systems.
  • gemcitabine 2 ⁇ M, 800 nM, 320 nM, 128 nM, 51.2 nM, 20.5 nM, 8.2 nM, and 0 nM
  • TGF-beta 2 antisense oligonucleotide according to Example 1 (0 ⁇ M ( ⁇
  • gemcitabine does not negatively influence the interaction of TGF-beta 2 antisense oligonucleotide with its TGF-beta 2 target, i.e., gemcitabine does not impair the suppression of TGF-beta 2 secretion via the TGF-beta 2 antisense oligonucleotide.
  • Example 5 TGF-beta 2 antisense oligonucleotide reducing the IC 50 of temozolomide (Fig. 5)
  • temozolomid concentrations i.e. 200 ⁇ M, 100 ⁇ M, 50 ⁇ M, 25 ⁇ M, 12.5 ⁇ M, 6.25 ⁇ M, 3.125 ⁇ M, and 0 ⁇ M temozolomide, respectively, in combination with 0 ⁇ M ( ⁇ ), 5 ⁇ M ( ⁇ ) or 10 ⁇ M (V) TGF-beta 2 antisense oligonucleotide, for example SEQ ID No. 30, for 2 days.
  • Substances (temozolomide and TGF-beta 2 antisense oligonucleotide) were prepared in aqueous solution, stored at 4°C and were used for 4 weeks.
  • the first treatment solution comprising temozolomidee and the TGF-beta 2 antisense oligonucleotide was removed and replaced by a second treatment solution containing the TGF-beta 2 antisense oligonucleotide and temozolomide in the above mentioned concentrations for further 2 days.
  • the proliferation/viability of the MEL-Juso cells was analyzed using the Cyquant method according to the manufacturer's instructions (Invitrogen), and the OD was measured after an incubation time of 60min with Cyquant solutions (detection reagent, direct nucleic acid stain, and direct nucleic acid background suppressor) using the plate reader "Fluostar- Optima" (BMG LABTECH GmbH).
  • Cyquant solutions detection reagent, direct nucleic acid stain, and direct nucleic acid background suppressor
  • TGF-beta 2 antisense oligonucleotide increasing temozolomide ' s cytotoxicity (Fig. 6)
  • A-172 cells (about 7000 cells/well) were seeded into 48-well plates, and 6 h after seeding 0 ⁇ M, 200 ⁇ M, or 800 ⁇ M temozolomide either alone (grey column) or in combination with 10 ⁇ M of a TGF-beta 2 antisense oligonuclotide (black column), for example SEQ ID No. 30, was added to the cells. After 2d of incubation, the treatment solutions were replaced and cells were incubated for additional 3d (total treatment time: 5d). Thereafter, cell supernatants containing lactate dehydrogenase (LDH) from lysed cells and of cells floating in the supernatant, as a result of treatment induced stress, were removed from the wells.
  • LDH lactate dehydrogenase
  • the cells of the supernatant were lysed by addition of lysis solution for example from the CytoToxicity Detection Kit Plus (Roche Diagnostics GmbH) and the LDH levels were determined for example according to the manual of the kit.
  • the amount of released LDH significantly increased with the combination of temozolomide and the TGF-beta 2 antisense oligonucleotide (Fig. 6).
  • Antisense m-RNA for the human TGF-beta 1 is an antisense m-RNA for the human TGF-beta 1 :
  • Antisense m-RNA of the human TGF-beta2 is an antisense m-RNA of the human TGF-beta2:
  • SB-431542 TBRI kinase inhibitor from GlaxoSmithKline (Callahan et al. 2002, Laping et al.
  • NPC30345 TBRI kinase inhibitor from Scios, Inc. (Dumont & Arteaga 2003)
  • TGF-B (Border et al. 1992).
  • Fc (RII/Fc hu IgGl fusion protein) from Biogen (Muraoka et al. 2002, Rowland- Goldsmith et al. 2001)
  • Betaglycan (recombinant soluble TBRIII) (Bandyopadhyay et al. 2002)
  • RXXR cleavage site of the mature (active) part (XX may be anything)
  • ASPC the C of this motif is the C for the intermolecular cystine bridge that links the two monomers into a functional dimer
  • mature protein of TGF-beta 1, 2 and 3 contains 112 amino acids from the end of this listing
  • TGF-beta3 YLWSSDTQHSRVLSLYNTINPEASASPCCVSQDLEPLTILYYIGKTPKIEQLSNMIVKSCKCS preferred amino acid sequences of TGF-beta3 :
  • a CACACAGT 14 1 A CACACAGT 14 1
  • GACCGTGG 18 192 GF- ⁇ -3rwk- CAAAGTTC 6
  • CAACTGGA 18 197 GF- ⁇ -3rwk- CAGAGA 11
  • ATGGATTT 17 201 GF- ⁇ -3rwk- CTTTGGCA 15
  • VEGF 90 CGGCCGCG 14 119 VEGF-98-1

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Epidemiology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Endocrinology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Peptides Or Proteins (AREA)
PCT/EP2010/061152 2009-07-30 2010-07-30 Combination of a chemotherapeutic agent and an inhibitor of the tgf-beta system WO2011012713A1 (en)

Priority Applications (11)

Application Number Priority Date Filing Date Title
IN969DEN2012 IN2012DN00969A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html) 2009-07-30 2010-07-30
US13/059,623 US8476246B2 (en) 2009-07-30 2010-07-30 Combination of a chemotherapeutic agent and an inhibitor of the TGF-beta system
RU2012107535/15A RU2012107535A (ru) 2009-07-30 2010-07-30 КОМБИНАЦИЯ ХИМИОТЕРАПЕВТИЧЕСКОГО СРЕДСТВА И ИНГИБИТОРА СИСТЕМЫ TGF-β
MX2012001244A MX2012001244A (es) 2009-07-30 2010-07-30 Combinacion de un agente quimioterapeutico y un inhibidor del sistema tgf-beta.
CA2769618A CA2769618A1 (en) 2009-07-30 2010-07-30 Combination of a chemotherapeutic agent and an inhibitor of the tgf-beta system
EP10737580.0A EP2340308B1 (en) 2009-07-30 2010-07-30 Combination of dacarbazine and an inhibitor of the TGF-beta system
AU2010277554A AU2010277554B2 (en) 2009-07-30 2010-07-30 Combination of a chemotherapeutic agent and an inhibitor of the TGF-beta system
BR112012002102A BR112012002102A2 (pt) 2009-07-30 2010-07-30 composição farmaceutica e metodo para preparar uma composição farmaceutica
CN2010800436805A CN102712924A (zh) 2009-07-30 2010-07-30 化疗药剂和TGF-β系统抑制剂的组合
JP2012522185A JP2013500313A (ja) 2009-07-30 2010-07-30 化学療法薬およびＴＧＦ−β系の阻害剤の組合せ
IL217821A IL217821A0 (en) 2009-07-30 2012-01-29 Combination of a chemotherapeutic agent and an inhibitor of the tgf-beta system

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP09166893 2009-07-30
EP09166893.9 2009-07-30

Publications (1)

Publication Number Publication Date
WO2011012713A1 true WO2011012713A1 (en) 2011-02-03

Family

ID=41278403

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2010/061152 WO2011012713A1 (en) 2009-07-30 2010-07-30 Combination of a chemotherapeutic agent and an inhibitor of the tgf-beta system

Country Status (17)

Country Link
US (1) US8476246B2 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
EP (1) EP2340308B1 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
JP (1) JP2013500313A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
KR (1) KR20120107456A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
CN (1) CN102712924A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
AU (1) AU2010277554B2 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
BR (1) BR112012002102A2 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
CA (1) CA2769618A1 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
CL (1) CL2012000247A1 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
CO (1) CO6511199A2 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
EC (1) ECSP12011689A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
IL (1) IL217821A0 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
IN (1) IN2012DN00969A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
MX (1) MX2012001244A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
PE (1) PE20121495A1 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
RU (1) RU2012107535A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)
WO (1) WO2011012713A1 (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014159774A1 (en) * 2013-03-14 2014-10-02 Teva Pharmaceutical Industries Ltd. Custirsen treatment with reduced toxicity
WO2017138924A1 (en) 2016-02-09 2017-08-17 Autotelic Llc Compositions and methods for treating pancreatic cancer
US9758786B2 (en) 2016-02-09 2017-09-12 Autotelic, Llc Compositions and methods for treating pancreatic cancer
JP2022535102A (ja) * 2019-06-05 2022-08-04 アイオー バイオテック エーピーエス Ｔｇｆ－ベータワクチン

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE402999T1 (de) * 2002-01-17 2008-08-15 Univ British Columbia Bispezifische antisense oligonukleotide die igfbp-2 und igfbp-5 inhibieren und deren verwendung
CN103517990A (zh) 2010-10-07 2014-01-15 通用医疗公司 癌症生物标志物
TWI454281B (zh) * 2011-10-06 2014-10-01 Nat Univ Tsing Hua 硼酸於硼中子捕獲治療肝癌之用途
WO2013124869A2 (en) * 2012-02-21 2013-08-29 Amrita Vishwa Vidyapeetham University The art, method,manner process and system of fibrous bio-degradable polymeric wafers for the local delivery of therapeutic agents in combinations
CA2949574C (en) * 2014-05-23 2022-06-07 Inserm (Institut National De La Sante Et De La Recherche Medicale) Methods for determining whether a patient will achieve a response after radiation therapy
US10301624B2 (en) 2014-06-25 2019-05-28 The General Hospital Corporation Targeting human satellite II (HSATII)
US11759530B2 (en) * 2014-11-17 2023-09-19 City Of Hope TKI permeability enhancers
US20180161429A1 (en) 2015-06-26 2018-06-14 Beth Israel Deaconess Medical Center Inc. Cancer therapy targeting tetraspanin 33 (tspan33) in myeloid derived suppressor cells
WO2017138925A1 (en) 2016-02-09 2017-08-17 Autotelic Llc Antitumour combinations of antisense oligonucleotides and anticancer agents
CA3037046A1 (en) 2016-10-31 2018-05-03 University Of Massachusetts Targeting microrna-101-3p in cancer therapy
WO2018089688A1 (en) 2016-11-09 2018-05-17 Jinjun Shi Restoration of tumor suppression using mrna-based delivery system
CN112533613A (zh) 2018-02-06 2021-03-19 通用医疗公司 作为肿瘤免疫应答的生物标志物的重复rna
KR20210139237A (ko) * 2019-03-14 2021-11-22 더 리서치 파운데이션 포 더 스테이트 유니버시티 오브 뉴욕 변형된 마이크로rna 및 암 치료에 있어서의 이의 용도
CN112933238A (zh) * 2021-02-02 2021-06-11 石河子大学 抑制TGF-βR1基因表达的物质与顺铂在治疗和/或预防食管癌中的应用
CN113425855B (zh) * 2021-06-28 2023-11-07 深圳瑞吉生物科技有限公司 一种mRNA剂型的骨关节炎药物制剂及其制备方法和应用

Citations (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3854480A (en) 1969-04-01 1974-12-17 Alza Corp Drug-delivery system
EP0092574A1 (en) 1981-10-23 1983-11-02 Molecular Biosystems Inc OLIGONUCLEOTIDE THERAPEUTIC AGENT AND METHODS OF MAKING SAME.
US4452775A (en) 1982-12-03 1984-06-05 Syntex (U.S.A.) Inc. Cholesterol matrix delivery system for sustained release of macromolecules
US4469863A (en) 1980-11-12 1984-09-04 Ts O Paul O P Nonionic nucleic acid alkyl and aryl phosphonates and processes for manufacture and use thereof
US4675189A (en) 1980-11-18 1987-06-23 Syntex (U.S.A.) Inc. Microencapsulation of water soluble active polypeptides
US5023243A (en) 1981-10-23 1991-06-11 Molecular Biosystems, Inc. Oligonucleotide therapeutic agent and method of making same
US5075109A (en) 1986-10-24 1991-12-24 Southern Research Institute Method of potentiating an immune response
US5133974A (en) 1989-05-05 1992-07-28 Kv Pharmaceutical Company Extended release pharmaceutical formulations
US5407686A (en) 1991-11-27 1995-04-18 Sidmak Laboratories, Inc. Sustained release composition for oral administration of active ingredient
US5539082A (en) 1993-04-26 1996-07-23 Nielsen; Peter E. Peptide nucleic acids
US5714331A (en) 1991-05-24 1998-02-03 Buchardt, Deceased; Ole Peptide nucleic acids having enhanced binding affinity, sequence specificity and solubility
US5719262A (en) 1993-11-22 1998-02-17 Buchardt, Deceased; Ole Peptide nucleic acids having amino acid side chains
US5736152A (en) 1995-10-27 1998-04-07 Atrix Laboratories, Inc. Non-polymeric sustained release delivery system
WO2002017852A2 (en) 2000-08-25 2002-03-07 Genta Incorporated Methods of treatment of a bcl-2 disorder using bcl-2 antisense oligomers
WO2005059133A2 (en) 2003-12-19 2005-06-30 Antisense Pharma Gmbh Combination therapy associating a tgf-breta antagonist with a chemiotherapeutic agent

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0856579A1 (en) * 1997-01-31 1998-08-05 BIOGNOSTIK GESELLSCHAFT FÜR BIOMOLEKULARE DIAGNOSTIK mbH An antisense oligonucleotide preparation method
US6809118B2 (en) * 2002-07-25 2004-10-26 Yih-Lin Chung Methods for therapy of radiation cutaneous syndrome
JP4682312B2 (ja) * 2003-09-04 2011-05-11 学校法人日本大学 ＴＧＦ−β遺伝子発現抑制剤
JP2007515949A (ja) * 2003-11-13 2007-06-21 ジェネンテック・インコーポレーテッド スクリーニングアッセイ及び腫瘍治療の方法
CN1902488A (zh) * 2003-11-13 2007-01-24 健泰科生物技术公司 肿瘤治疗的筛选实验和方法
US20070155685A1 (en) * 2004-02-27 2007-07-05 Karl-Hermann Schlingensiepen Pharmaceutical composition
US20090186076A1 (en) * 2006-02-01 2009-07-23 Kazunori Kataoka Combined Use of TGF-Beta Signaling Inhibitor and Antitumor Agent
JP5322476B2 (ja) * 2008-03-31 2013-10-23 テルモ株式会社 リポソームの製造装置およびリポソームの製造方法

Patent Citations (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3854480A (en) 1969-04-01 1974-12-17 Alza Corp Drug-delivery system
US4469863A (en) 1980-11-12 1984-09-04 Ts O Paul O P Nonionic nucleic acid alkyl and aryl phosphonates and processes for manufacture and use thereof
US4675189A (en) 1980-11-18 1987-06-23 Syntex (U.S.A.) Inc. Microencapsulation of water soluble active polypeptides
EP0092574A1 (en) 1981-10-23 1983-11-02 Molecular Biosystems Inc OLIGONUCLEOTIDE THERAPEUTIC AGENT AND METHODS OF MAKING SAME.
US5023243A (en) 1981-10-23 1991-06-11 Molecular Biosystems, Inc. Oligonucleotide therapeutic agent and method of making same
US4452775A (en) 1982-12-03 1984-06-05 Syntex (U.S.A.) Inc. Cholesterol matrix delivery system for sustained release of macromolecules
US5075109A (en) 1986-10-24 1991-12-24 Southern Research Institute Method of potentiating an immune response
US5133974A (en) 1989-05-05 1992-07-28 Kv Pharmaceutical Company Extended release pharmaceutical formulations
US5714331A (en) 1991-05-24 1998-02-03 Buchardt, Deceased; Ole Peptide nucleic acids having enhanced binding affinity, sequence specificity and solubility
US5407686A (en) 1991-11-27 1995-04-18 Sidmak Laboratories, Inc. Sustained release composition for oral administration of active ingredient
US5539082A (en) 1993-04-26 1996-07-23 Nielsen; Peter E. Peptide nucleic acids
US5719262A (en) 1993-11-22 1998-02-17 Buchardt, Deceased; Ole Peptide nucleic acids having amino acid side chains
US5736152A (en) 1995-10-27 1998-04-07 Atrix Laboratories, Inc. Non-polymeric sustained release delivery system
WO2002017852A2 (en) 2000-08-25 2002-03-07 Genta Incorporated Methods of treatment of a bcl-2 disorder using bcl-2 antisense oligomers
WO2005059133A2 (en) 2003-12-19 2005-06-30 Antisense Pharma Gmbh Combination therapy associating a tgf-breta antagonist with a chemiotherapeutic agent
US20070196269A1 (en) * 2003-12-19 2007-08-23 Karl-Hermann Schlingensiepen Pharmaceutical composition

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014159774A1 (en) * 2013-03-14 2014-10-02 Teva Pharmaceutical Industries Ltd. Custirsen treatment with reduced toxicity
WO2017138924A1 (en) 2016-02-09 2017-08-17 Autotelic Llc Compositions and methods for treating pancreatic cancer
US9758786B2 (en) 2016-02-09 2017-09-12 Autotelic, Llc Compositions and methods for treating pancreatic cancer
US9963703B2 (en) 2016-02-09 2018-05-08 Autotelic Llc Compositions and methods for treating pancreatic cancer
JP2022535102A (ja) * 2019-06-05 2022-08-04 アイオー バイオテック エーピーエス Ｔｇｆ－ベータワクチン

Also Published As

Publication number Publication date
EP2340308A1 (en) 2011-07-06
US8476246B2 (en) 2013-07-02
CL2012000247A1 (es) 2013-07-05
CO6511199A2 (es) 2012-08-31
BR112012002102A2 (pt) 2017-05-02
IN2012DN00969A (GUID-C5D7CC26-194C-43D0-91A1-9AE8C70A9BFF.html) 2015-04-10
EP2340308B1 (en) 2014-12-24
MX2012001244A (es) 2012-03-26
PE20121495A1 (es) 2012-11-19
IL217821A0 (en) 2012-03-29
RU2012107535A (ru) 2013-09-10
KR20120107456A (ko) 2012-10-02
JP2013500313A (ja) 2013-01-07
US20120027873A1 (en) 2012-02-02
CN102712924A (zh) 2012-10-03
AU2010277554A1 (en) 2012-03-01
AU2010277554B2 (en) 2015-02-19
CA2769618A1 (en) 2011-02-03
ECSP12011689A (es) 2012-06-29

Similar Documents

Publication Publication Date Title
AU2010277554B2 (en) Combination of a chemotherapeutic agent and an inhibitor of the TGF-beta system
AU2004299670B2 (en) Combination therapy associating a TGF-beta antagonist with a chemotherapeutic agent
US20030055014A1 (en) Inhibition of angiogenesis by nucleic acids
US20040097463A1 (en) Use of asiatic acid or asiaticoside for treatment of cancer
US12303494B2 (en) Compositions and methods for monitoring, diagnosis, prognosis, detection, and treatment of cancer
US20090143418A1 (en) Compositions and Methods for Preventing and Treating Hair Growth Cycle-Related Conditions
CN117343965B (zh) 核酸载体、核酸药及其制备方法和应用
JP5667043B2 (ja) 阻害性オリゴヌクレオチドを送達するための組成物および方法
US7235583B1 (en) Fatty acid-anticancer conjugates and uses thereof
US20050267027A1 (en) Use of erythropoietin for treatment of cancer
JP2022506958A (ja) Ｍｉｒ－１０ｂ活性を調節するためのマイクロｒｎａ化合物及び方法
EP2248895A2 (en) Combination therapy associating a TGF-beta antagonist with a chemotherapeutic agent
US11547718B2 (en) Modulators of FOXP3 expression
WO2020160453A1 (en) Modulators of yap1 expression
US20200181711A1 (en) Compositions and methods for monitoring, diagnosis, prognosis, detection, and treatment of cancer
US20220168422A1 (en) Compositions and methods for monitoring, diagnosis, prognosis, detection, and treatment of cancer
US20220233539A1 (en) Compositions and methods for inhibiting hedgehog pathway
HK40059024A (en) Modulators of foxp3 expression

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 201080043680.5

Country of ref document: CN

WWE Wipo information: entry into national phase

Ref document number: 2010737580

Country of ref document: EP

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10737580

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 13059623

Country of ref document: US

WWE Wipo information: entry into national phase

Ref document number: 2012522185

Country of ref document: JP

Ref document number: MX/A/2012/001244

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 217821

Country of ref document: IL

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2012000247

Country of ref document: CL

Ref document number: 2769618

Country of ref document: CA

Ref document number: 000130-2012

Country of ref document: PE

WWE Wipo information: entry into national phase

Ref document number: 969/DELNP/2012

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 2010277554

Country of ref document: AU

ENP Entry into the national phase

Ref document number: 20127005337

Country of ref document: KR

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 2012107535

Country of ref document: RU

Ref document number: 12035227

Country of ref document: CO

ENP Entry into the national phase

Ref document number: 2010277554

Country of ref document: AU

Date of ref document: 20100730

Kind code of ref document: A

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112012002102

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 112012002102

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20120130