[go: up one dir, main page]

WO2012110425A1 - Procédés pour surveiller la réponse au traitement et pour traiter le cancer colorectal - Google Patents

Procédés pour surveiller la réponse au traitement et pour traiter le cancer colorectal Download PDF

Info

Publication number
WO2012110425A1
WO2012110425A1 PCT/EP2012/052345 EP2012052345W WO2012110425A1 WO 2012110425 A1 WO2012110425 A1 WO 2012110425A1 EP 2012052345 W EP2012052345 W EP 2012052345W WO 2012110425 A1 WO2012110425 A1 WO 2012110425A1
Authority
WO
WIPO (PCT)
Prior art keywords
calpain
cancer
cells
inhibitor
expression
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/EP2012/052345
Other languages
English (en)
Inventor
Jean-François PEYRON
Patricia LAGADEC
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Institut National de la Sante et de la Recherche Medicale INSERM
Original Assignee
Institut National de la Sante et de la Recherche Medicale INSERM
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
Application filed by Institut National de la Sante et de la Recherche Medicale INSERM filed Critical Institut National de la Sante et de la Recherche Medicale INSERM
Publication of WO2012110425A1 publication Critical patent/WO2012110425A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
    • 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
    • A61K31/65—Tetracyclines
    • 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
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/06—Tripeptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/07—Tetrapeptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/55—Protease inhibitors
    • A61K38/57—Protease inhibitors from animals; from humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • the present invention relates to methods for monitoring the response to treatment and for treating colorectal cancer.
  • CRC Colorectal cancer
  • CPT-l l/SN-38 is a semisynthetic analog of camptothecin. It interacts with cellular topoisomerase I-DNA complexes (Topo I) which results in induction of a S -phase- specific cytotoxicity (5).
  • CPT-11 is a pro-drug which is converted into the active molecule SN-38 by carboxylesterases and a clear relationship between carboxylesterase level and the chemosensitivity of human lung cancer cells has been demonstrated in vitro.
  • the mechanisms of CPT-l l/SN-38 resistance have not been fully understood [reviewed in (6)]. It has been reported that the efflux of CPT-11 or SN-38 is due to the overexpression of the ATP-binding cassette (ABC) transmembrane transporters such as P-glycoprotein (P-gp) (7) and the breast cancer protein (BCRP) (8).
  • ABSC ATP-binding cassette
  • P-gp P-glycoprotein
  • BCRP breast cancer protein
  • tumor cells may escape irinotecan cytotoxic effects by reducing the level of Topo I expression (9).
  • NF-KB nuclear factor kappa B
  • IKK ⁇ kinase
  • IKK2/B NEMO (NF- ⁇ Essential Modulator)/IKKy, form the IKK complex that integrates signals for NF- ⁇ activation.
  • the invention relates to a method for delaying or inhibiting the secondary resistance to an anti-cancer agent, particularly CPT-l l/SN-38, in a patient suffering from colorectal cancer, comprising the administration of a calpain inhibitor to said patient.
  • the present invention also relates to a calpain inhibitor for use for delaying or inhibiting the secondary resistance to an anti-cancer agent, particularly CPT-l l/SN-38, in a patient suffering from colorectal cancer.
  • a product containing an anti-cancer agent, particularly CPT-l l/SN-38, and a calpain inhibitor as a combined preparation for simultaneous, separate or sequential use in colorectal cancer therapy.
  • the present invention also relates to a method for monitoring the response to a treatment of a patient suffering from a cancer chosen from colorectal cancer, gastric cancer, esophageal cancer, non-small-cell and small-cell lung cancers, leukemia, lymphomas, central nervous system maligant gliomas and ovarian carcinomas, comprising:
  • said patient is suffering from colorectal cancer.
  • calpain inhibitor refers to compounds which inhibit signalling through calpain 1 (calpain 1 inhibitors) or calpain 2 (calpain 2 inhibitors), as well as compounds which inhibit the expression of the calpain 1 or calpain 2 genes.
  • Compounds which inhibit signalling through calpain 1 or calpain 2 are called calpain antagonists. They include compounds which inhibit the activity of calpain 1 or calpain 2, by binding to said calpain, or by inhibiting calpain 1 or calpain 2 signalling by other mechanisms.
  • Said calpain antagonists may be chosen from specific calpain 1 antagonists and specific calpain 2 antagonists.
  • specific or “selective” it is meant that the affinity of the antagonist for said calpain (i.e. calpain 1 or calpain 2) is at least 10-fold, preferably 25- fold, more preferably 100-fold, still preferably 500-fold higher than the affinity for the other calpains.
  • Said calpain antagonists may also be chosen from specific calpain 1/2 antagonists.
  • specific calpain 1/2 antagonists it is meant that the affinity of the antagonist for said calpains (i.e. calpain 1 and calpain 2) is at least 10-fold, preferably 25-fold, more preferably 100-fold, still preferably 500-fold higher than the affinity for the other calpains.
  • calpain antagonists are synthetic compounds such as ALLN (calpain inhibitor I) and ALLM (calpain inhibitor II) peptides developed by Peptides International.
  • ALLN is Acetyl-L-Leucyl-L-Leucyl-L-Norleucinal (N-Ac-Leu-Leu-Nle-CHO (aldehyde)), and inhibits preferentially calpain 1, but also calpain 2 to a less extent. It is also a proteasome and cathepsin inhibitor.
  • ALLM is Acetyl-L-Leucyl-L-Leucyl-L-Methioninal (N-Ac-Leu-Leu-Met-CHO (aldehyde)), and inhibits preferentially calpain 1, but also calpain 2 to a less extent. It is also a cathepsin inhibitor.
  • Calpain antagonists also include:
  • calpastatin which is encoded by the CAST gene and which is the natural antagonist of calpains 1 and 2 (Paquet-Durand F et al., Journal of neurochemistry, 115:930-940, 2010);
  • calpeptin also known as benzyloxycarbonyldipeptidyl aldehyde
  • MDL28170 and SJA6017 and A-705253 (N-( l -ben/y!-2-carbamoyl- 2-oxoethyl )-2-(H-2-(4-cliethylaminomethylp enyl )ethen- 1 -v! )ben/amicle).
  • SNJ- 1945 is also known as 2-(2-methoxyethoxy)ethyl N-[(1S)-1-[[(2S)-1- (cyclopropylcarbamoyl)-l-oxo-3-phenyl-propan-2-yl]carbamoyl]-3-methyl- butyl] carbamate, which is an orally available calpain inhibitor.
  • SJA6017 is also known as N-(4-Fluorophenylsulfonyl)-L-Valyl-L-Leucinal ; AK295, also known as Cbz-Leu-aminobutyrate-CONH(CH 2 )3 morpholine; and tetracyclines, like chlortetracycline (CTC) and demeclocycline (DMC).
  • CTC chlortetracycline
  • DMC demeclocycline
  • Calpain inhibitors also include compounds which inhibit the expression of the calpain 1 or calpain 2 genes; said compounds are called inhibitors of calpain gene expression.
  • An "inhibitor of gene expression" refers to a natural or synthetic compound that has a biological effect to inhibit or significantly reduce the expression of said gene. Consequently an inhibitor of calpain 1 or calpain 2 gene expression refers to a natural or synthetic compound that has a biological effect to inhibit or significantly reduce the expression of the calpain 1 or calpain 2 genes respectively.
  • the inhibitors of calpain gene expression include, but are not limited to, antisense oligonucleotides, siRNAs, shRNAs, ribozymes and DNAzymes.
  • said inhibitor of calpain gene expression is chosen from siRNAs.
  • colon cancer refers to the pathological condition in mammals that is typically characterized by unregulated cell growth in the colon, rectum and appendix. It is also called colon cancer or large bowel cancer.
  • a patient denotes a mammal, such as a rodent, a feline, a canine, and a primate.
  • a patient according to the invention is a human.
  • treating means reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or reversing, alleviating, inhibiting the progress of, or preventing one or more symptoms of the disorder or condition to which such term applies.
  • anti-cancer agent or “chemotherapeutic agent” refers to compounds which are used in the treatment of colorectal cancer.
  • Anti-cancer agents include but are not limited to fludarabine, gemcitabine, capecitabine, methotrexate, taxol, taxotere, mercaptopurine, thioguanine, hydroxyurea, cytarabine, cyclophosphamide, ifosfamide, nitrosoureas, platinum complexes such as cisplatin, carboplatin and oxaliplatin, mitomycin, dacarbazine, procarbizine, etoposide, teniposide, campathecins, bleomycin, doxorubicin, idarubicin, daunorubicin, dactinomycin, plicamycin, mitoxantrone, L-asparaginase, doxorubicin, epimbicm, 5- fluorouracil, taxanes such as docetaxel and paclitaxel, leucovorin, levamisole, irinote
  • the anti-cancer agent is irinotecan or SN-38.
  • the expression “secondary resistance to an anti-cancer agent”, or “acquired resistance to an anti-cancer agent”, refers to the resistance to both apoptosis and antiproliferative effect that occurs during the treatment, and that is induced by said anti-cancer agent in a patient suffering from colorectal cancer, said patient being an initial responder (sensitive) to said treatment.
  • the secondary resistance appears in almost all patients who initially responded to the first line chemotherapy with said anticancer agent, particularly with CPT-11 or SN-38; the colorectal cancerous cells of the patient are then less or no more sensitive to the anti-cancer agent.
  • Secondary resistance to an anti-cancer agent has to be distinguished from primary resistance (or innate resistance), which is the resistance to both apoptosis and antiproliferative effect that occurs in a patient suffering from colorectal cancer at the beginning of the treatment.
  • primary resistance or innate resistance
  • said patient is an initial non responder (not sensitive) to said treatment.
  • the present invention relates to a calpain inhibitor for use for delaying or inhibiting the secondary resistance to an anti-cancer agent in a patient suffering from colorectal cancer.
  • the invention also relates to a method for delaying or inhibiting the secondary resistance to an anti-cancer agent in a patient suffering from colorectal cancer, comprising the administration of a calpain inhibitor to said patient.
  • the invention relates to the use of a calpain inhibitor for the manufacture of a medicament for delaying or inhibiting the secondary resistance to an anti-cancer agent in a patient suffering from colorectal cancer.
  • the calpain inhibitor is used for delaying or inhibiting the secondary resistance to irinotecan or SN-38 in a patient suffering from colorectal cancer.
  • Said calpain inhibitor is preferably a calpain 2 inhibitor.
  • delaying the secondary resistance to an anti-cancer agent it is meant postponing the onset of secondary resistance to said anti-cancer agent in a patient.
  • inhibiting the secondary resistance to an anti-cancer agent it is meant preventing the onset of secondary resistance to said anti-cancer agent in a patient.
  • the calpain inhibitor is used in combination with chemotherapy, i.e. in combination with an anti-cancer agent.
  • the anti-cancer agent is selected from 5-fluorouracil, bevacizumab, irinotecan, SN38, bortezomib, oxaliplatin, cetuximab, panitumumab, leucovorine and capecitabine.
  • the anti-cancer agent is selected from irinotecan and SN38.
  • the present invention thus also relates to a product containing an anti-cancer agent and a calpain inhibitor as a combined preparation for simultaneous, separate or sequential use in colorectal cancer therapy.
  • the present invention relates to a method for treating colorectal cancer in a secondary resistant patient comprising administering a therapeutically effective amount of a calpain inhibitor and a therapeutically effective amount of an anti-cancer agent.
  • medicaments according to the invention comprise a calpain inhibitor, and optionally an anti-cancer agent, together with a pharmaceutically-acceptable carrier.
  • suitable carriers Suitable formulations for administration by any desired route may be prepared by standard methods, for example by reference to well-known text such as Remington; The Science and Practice of Pharmacy.
  • HT-29R resistant tumor
  • high levels of expression of said genes in the colon cancer cells of a patient are indicative of a secondary resistance to treatment with an anti-cancer agent, particularly CPT-l l/SN- 38, of said patient compared to patients who do not express or express low levels of said genes in said colon cancer cells.
  • an anti-cancer agent particularly CPT-l l/SN- 38
  • cancers like gastric cancer, esophageal cancer, non- small-cell and small-cell lung cancers, leukemia, lymphomas, central nervous system maligant gliomas and ovarian carcinomas, may be treated with CPT-11.
  • This compound has indeed been shown as clinically active against these cancers (Rothenberg ML et al., 2001; 6(1), 66- 80; and (4)).
  • the invention thus provides a method for monitoring the response to a treatment of a patient suffering from a cancer chosen from colorectal cancer, gastric cancer, esophageal cancer, non- small-cell and small-cell lung cancers, leukemia, lymphomas, central nervous system maligant gliomas and ovarian carcinomas, and preferably colorectal cancer, comprising:
  • high levels of expression of calpain, annexin A2 and S 100A10 genes, particularly of at least calpain 2 gene, in colon cancer cells is indicative of secondary resistance to treatment with an anti-cancer agent.
  • the inventors have demonstrated in the example that calpain 2, S 100A10 and annexin A2 are overexpressed in resistant colon cancer cells, and that calpain 2 overexpression is involved in secondary resistance to CPT-l l/SN-38 treatment.
  • the anti-cancer agent is as defined above.
  • the anti-cancer agent is selected from CPT-11 and SN-38.
  • Step a comprises preferably the treatment of said patient with the anti-cancer agent during at least 5 months, preferably at least 6 months.
  • step b. of the method according to the invention comprises measuring the level of expression of at least the calpain 2 gene in cancer cells of said patient.
  • step b. of the method according to the invention comprises measuring the level of expression of at least S100A10 and annexin A2 genes in cancer cells of said patient.
  • step b. of the method according to the invention comprises measuring the levels of expression of all the calpain, annexin A2 and S IOOAIO genes, in cancer cells of said patient.
  • step b. of the method according to the invention comprises measuring the levels of expression of all the calpain 2, annexin A2 and S IOOAIO genes, in cancer cells of said patient.
  • Calpain genes refer to calpain 1 gene and calpain 2 gene.
  • Annexin A2 also known as annexin II, is a protein that in humans in encoded by the ANXA2 gene. It is a member of the annexin family, and it has interactions with some ligands, including S IOOAIO.
  • S IOOAIO also known as pi 1
  • pi 1 is a protein that is encoded by the S IOOAIO gene in humans, and the S IOOAIO gene in other species. It is a member of the SlOO family of proteins containing two EF-hand calcium binding motifs, and it is not calcium- dependent.
  • the term "gene expression level” or “level of expression of a gene” refers to an amount or a concentration of a transcription product, for instance mRNA, or of a translation product, for instance a protein or polypeptide.
  • a level of mRNA expression can be expressed in units such as transcripts per cell or nanograms per microgram of tissue.
  • a level of a polypeptide can be expressed as nanograms per microgram of tissue or nanograms per milliliter of a culture medium, for example.
  • relative units can be employed to describe a gene expression level.
  • the expression of "measuring the level of expression of a gene” encompasses the step of measuring the quantity of a transcription product, preferably mRNA obtained through transcription of said gene, and/or the step of measuring the quantity of translation product, preferably the protein obtained through translation of said gene.
  • the step of measuring the expression of a gene refers to the step of measuring the quantity of mRNA obtained through transcription of said gene.
  • step b. of measuring the gene expression level is performed by the following method: a) obtaining a biological sample comprising cancer cells, preferably colon cancer cells, from said patient,
  • a biological sample in case of monitoring the response to a treatment of a patient suffering from colorectal cancer, may be a sample of the colorectal tumor tissue or colon cancer cells obtained from the patient according to methods known in the art. Said biological sample is for example a biopsy.
  • step b. of measuring the gene expression level may be performed according to the routine techniques, well known of the person skilled in the art.
  • the measurement comprises contacting the cancer cells of the biological sample with selective reagents such as probes, primers, ligands or antibodies, and thereby detecting the presence of nucleic acids or proteins of interest originally in the sample.
  • selective reagents such as probes, primers, ligands or antibodies
  • the expression may be measured by measuring the level of mRNA.
  • nucleic acid contained in the samples e.g., isolated cancer cells prepared from the patient, like those included in biopsies
  • the extracted mRNA is then detected by hybridization (e. g., Northern blot analysis) and/or amplification (e.g., RT- PCR).
  • the expression of the calpain, annexin A2 or S 100A10 is measured by RT-PCR, preferably quantitative or semi-quantitative RT- PCR, even more preferably real-time quantitative or semi-quantitative RT-PCR.
  • nucleic acids having at least 10 nucleotides and exhibiting sequence complementarity or homology to the mRNA of interest herein find utility as hybridization probes or amplification primers. It is understood that such nucleic acids need not be identical, but are typically at least about 80% identical to the homologous region of comparable size, more preferably 85% identical and even more preferably 90-95% identical. In certain embodiments, it will be advantageous to use nucleic acids in combination with appropriate means, such as a detectable label, for detecting hybridization. A wide variety of appropriate indicators are known in the art including, fluorescent, radioactive, enzymatic or other ligands (e. g. avidin/biotin).
  • Probes typically comprise single- stranded nucleic acids of between 10 to 1000 nucleotides in length, for instance of between 10 and 800, more preferably of between 15 and 700, typically of between 20 and 500.
  • Primers typically are shorter single- stranded nucleic acids, of between 10 to 25 nucleotides in length, designed to perfectly or almost perfectly match a nucleic acid of interest, to be amplified.
  • the probes and primers are "specific" to the nucleic acids they hybridize to, i.e. they preferably hybridize under high stringency hybridization conditions (corresponding to the highest melting temperature Tm, e.g., 50 % formamide, 5x or 6x SCC.
  • SCC is a 0.15 M NaCl, 0.015 M Na-citrate).
  • the nucleic acid primers or probes used in the above amplification and detection method may be assembled as a kit.
  • a kit includes consensus primers and molecular probes.
  • a preferred kit also includes the components necessary to determine if amplification has occurred.
  • the kit may also include, for example, PCR buffers and enzymes; positive control sequences, reaction control primers; and instructions for amplifying and detecting the specific sequences.
  • the methods of the invention comprise contacting the cancer cells of the biological sample with a binding partner capable of selectively interacting with the calpain, annexin A2 or S 100A10 proteins present in the biological sample.
  • the binding partner may be an antibody that may be polyclonal or monoclonal, preferably monoclonal.
  • the binding partner may be an aptamer.
  • Polyclonal antibodies of the invention or a fragment thereof can be raised according to known methods by administering the appropriate antigen or epitope to a host animal selected, e.g., from pigs, cows, horses, rabbits, goats, sheep, and mice, among others.
  • Various adjuvants known in the art can be used to enhance antibody production.
  • antibodies useful in practicing the invention can be polyclonal, monoclonal antibodies are preferred.
  • Monoclonal antibodies of the invention or a fragment thereof can be prepared and isolated using any technique that provides for the production of antibody molecules by continuous cell lines in culture.
  • Techniques for production and isolation include but are not limited to the hybridoma technique originally described by Kohler and Milstein (1975); the human B-cell hybridoma technique (Cote et al., 1983); and the EBV- hybridoma technique (Cole et al. 1985).
  • Antibodies useful in practicing the present invention also include anti-calpain fragments and anti-annexin A2 fragments and anti- S 100A10 fragments including but not limited to F(ab')2 fragments, which can be generated by pepsin digestion of an intact antibody molecule, and Fab fragments, which can be generated by reducing the disulfide bridges of the F(ab')2 fragments.
  • Fab and/or scFv expression libraries can be constructed to allow rapid identification of fragments having the desired specificity to calpain, annexin A2 or S 100A10.
  • phage display of antibodies may be used.
  • single-chain Fv (scFv) or Fab fragments are expressed on the surface of a suitable bacteriophage, e. g., M13.
  • spleen cells of a suitable host e. g., mouse
  • the coding regions of the VL and VH chains are obtained from those cells that are producing the desired antibody against the protein. These coding regions are then fused to a terminus of a phage sequence.
  • the phage displays the antibody fragment.
  • Phage display of antibodies may also be provided by combinatorial methods known to those skilled in the art.
  • Antibody fragments displayed by a phage may then be used as part of an immunoassay.
  • the binding partner may be an aptamer.
  • Aptamers are a class of molecule that represents an alternative to antibodies in term of molecular recognition.
  • Aptamers are oligonucleotide or oligopeptide sequences with the capacity to recognize virtually any class of target molecules with high affinity and specificity.
  • Such ligands may be isolated through Systematic Evolution of Ligands by Exponential enrichment (SELEX) of a random sequence library, as described in Tuerk C. 1997.
  • the random sequence library is obtainable by combinatorial chemical synthesis of DNA. In this library, each member is a linear oligomer, eventually chemically modified, of a unique sequence.
  • Peptide aptamers consist of conformationally constrained antibody variable regions displayed by a platform protein, such as E. coli Thioredoxin A, that are selected from combinatorial libraries by two hybrid methods (Colas et al., 1996).
  • binding partners of the invention such as antibodies or aptamers, may be labelled with a detectable molecule or substance, such as a fluorescent molecule, a radioactive molecule or any others labels known in the art.
  • a detectable molecule or substance such as a fluorescent molecule, a radioactive molecule or any others labels known in the art.
  • Labels are known in the art that generally provide (either directly or indirectly) a signal.
  • the term "labelled", with regard to the antibody is intended to encompass direct labelling of the antibody or aptamer by coupling (i.e., physically linking) a detectable substance, such as a radioactive agent or a fluorophore (e.g. fluorescein isothiocyanate (FITC) or phycoerythrin (PE) or Indocyanine (Cy5)) to the antibody or aptamer, as well as indirect labelling of the probe or antibody by reactivity with a detectable substance.
  • a detectable substance such as a radioactive agent or a fluorophore (e.g. fluorescein isothiocyanate (FITC) or phycoerythrin (PE) or Indocyanine (Cy5))
  • FITC fluorescein isothiocyanate
  • PE phycoerythrin
  • Indocyanine Indocyanine
  • An antibody or aptamer of the invention may be labelled with a radioactive molecule by any method known in the art.
  • the aforementioned assays generally involve the binding of the binding partner (ie. antibody or aptamer) to a solid support.
  • Solid supports which can be used in the practice of the invention include substrates such as nitrocellulose (e. g., in membrane or microtiter well form); polyvinylchloride (e. g., sheets or microtiter wells); polystyrene latex (e.g., beads or microtiter plates); polyvinylidine fluoride; diazotized paper; nylon membranes; activated beads, magnetically responsive beads, and the like.
  • the gene expression level of the calpain, annexin A2 or SIOOAIO protein in cancer cells may be measured by using standard immunodiagnostic techniques, including immunoassays such as competition, direct reaction, or sandwich type assays.
  • immunoassays such as competition, direct reaction, or sandwich type assays.
  • cancers cells are purified from the isolated biological sample.
  • assays include, but are not limited to, agglutination tests; enzyme-labelled and mediated immunoassays, such as ELISAs; biotin/avidin type assays; radioimmunoassays; Immunoelectrophoresis ; immunoprecipitation.
  • an ELISA method can be used, wherein the wells of a microtiter plate are coated with a set of antibodies against the target (ie calpain, annexin A2 or S IOOAIO).
  • the cancer cells of the biological sample that are suspected of containing calpain, annexin A2 or S IOOAIO, are then added to the coated wells.
  • the plate(s) can be washed to remove unbound moieties and a detectably labelled secondary binding molecule added.
  • the secondary binding molecule is allowed to react with any captured sample marker protein, the plate washed and the presence of the secondary binding molecule detected using methods well known in the art.
  • A-B Study of HT-29 and HT-29R viability.
  • HT-29 and HT-9R cells were incubated for 5 days with AS602868, SN-38, both compounds simultaneously (A) or several concentrations of 5-FU or etoposide (B). Cytotoxicity was evaluated using the MTT assay. Data are expressed as mean + SD of quadruplicates of one representative experiment out of 5 (A) or 3 (B). Statistically significant differences between HT-29 and HT-29R cells are indicated on the figure (***p ⁇ 0.001).
  • HT-29 and HT-29R tumor-bearing mice received respectively CPT-11 i.p. injections (20 mg/ml) twice a week daily with (black squares) (black circles) or without (grey lozenges) (grey triangles) oral injections of AS602868 (20 mg/ml), 5 days a week. Data are the mean + SD of tumor measurements using 5 mice/group and are representative of 2 experiments. Statistically significant differences between CPT-11 treated HT-29 and HT-29R tumor-bearing mice and CPT-11 + AS602868 treated HT-29 and HT-29R tumor-bearing mice on the 10 th week are indicated on the figure.
  • A-B NF- ⁇ activation was visualized by EMS A.
  • A HT-29 and HT29R cells were treated with AS602868 30 min before stimulation with SN-38 for lh.
  • B Sensitive tumors (ST) were harvested from mice at the end of the first course of bitherapy (i.e. 10 th week). Resistant tumors (RT) were harvested at the end of the second course of treatment (i.e. 22 nd week). These results correspond to one representative experiment from 3.
  • C NF-KB activation was measured by reporter gene assay. Luciferase (RLU), Bgal activities and the protein concentration were measured in cell extracts. The luciferase activity was normalized and adjusted to 1 ⁇ g protein.
  • A Diagram showing the number of genes that are diferentially expressed between sensitive and resistant HT-29 cells and their expected involvement in the regulation of various biological processes through Ingenuity pathway program analysis.
  • B HT-29 and HT-29R (R) cells were analyzed for their RNA level of S 100A10, annexin A2 and calpain 2 by RT-PCR.
  • B actin served as an invariant loading control.
  • C HT-29, HT-29R, RAS, RSN and RAS+SN cells were analyzed for S 100A10, annexin A2 and calpain 2 expression by western blotting.
  • HSP60 served as an invariant loading control.
  • Sensitive and resistant tumors were analyzed for S100A10, annexin A2 and calpain 2 expression by western blotting. HSP60 served as an invariant loading control.
  • S 100A10, annexin A2 and calpain 2 expression was analyzed by western blotting after transient transfection of respective siRNA in HT-29R cells. Protein expression was observed after 3, 5 or 7 days of siRNA tranfection.
  • HT-29R cells transfected with non relevant siRNA served as negative control.
  • HSP60 served as an invariant loading control.
  • HT-29R and RSN cells transfected with non relevant siRNA served as negative control.
  • HSP60 served as an invariant loading control.
  • HT-29 and HT-29R viability were incubated for 5 days with growing concentrations of AS602868 or SN-38. Cytotoxicity was evaluated using the MTT assay. Data are expressed as mean + SD of quadruplicates of one representative experiment out of 5.
  • HSP60 served as an invariant loading control.
  • NF- ⁇ activation was visualized by EMSA.
  • HT-29R and RSN cells were treated with 10 ⁇ ALLN lh before harvesting.
  • AS602868 is an anilino-pyrimidine derivative and ATP competitor selected for its inhibitory effect in vitro on IKKee, a constitutively active version of IKK2.
  • AS602868 in sterile cyclodextrin solution as well as CPT-11 were supplied by Merck-Serono S.A. (Geneva, Switzerland).
  • SN-38 was a kind gift from Dr. J.L. Fischel from Antoine Lacassagne oncology center (Nice, France).
  • Etoposide and 5-fluorouracil were respectively obtained from Sigma Aldrich (Saint Quentin Fallavier, France) and MERCK generiques (Lyon, France).
  • Anti-procaspase 3 was purchased from Medical & Biological laboratories (Woburn, MA); anti-phospho- ⁇ from Cell Signaling (Beverly, MA); anti-HSP 60, anti- ⁇ , anti-Annexin A2 and anti-Calpain 2 from Santa Cruz Biotechnology (Santa Cruz, CA); anti S 100A10 from BD Biosciences (San Jose, CA).
  • the human colon cancer cell line HT-29 was obtained from the ATCC (Bethesda, MD).
  • the HT-29R cell line was established in our laboratory from a HT-29 xenograft resistant to CPT-11 plus AS602868 treatment.
  • RAS, RSN and RAS+SN cell lines were obtained by adding increasing concentrations of either AS602868 or SN-38 or both in the medium for 6 months.
  • RAS cells were grown in the presence of 3 ⁇ AS602868, RSN cells with 10 nM SN-38 and RAS+SN-38 cells with both.
  • Cytotoxic studies were carried out using a MTT assay (18), representing the percentage of viability inhibition induced by treatments.
  • Five hundred or one thousand HT-29 or HT-29R or RSN cells were respectively plated per well in 96-well plates with medium and various concentrations of AS602868 + SN-38 for 5 days.
  • Nucleic extracts of cells and tumors were prepared according to the method described by Dignam et al. (19). Nucleic extracts and EMSA were performed as described previously (10, 11). The density of bands was quantified using the ImageJ software (NIH, USA).
  • HT-29 and HT-29R cells were transfected using FuGENE (Roche) and 2 ⁇ g of a luciferase reporter gene controlled by a minimal thymidine kinase promoter with three reiterated ⁇ sites ( ⁇ 3 thymidine kinase luc).
  • FuGENE FuGENE
  • Detailed protocol has been previously published (11) Generation of stably tranfected cell lines
  • HT-29, HT-29R or RSN cells were transfected with 1 ⁇ g of mutated ⁇ - ⁇ cDNA (pcDNA3-Myc-huS3236AI-KB-a) a construct from our group, or empty vectors and 0.1 ⁇ g of the pBABE-puro plasmid (Addgene N°1764) which confers puromycin resistance using FuGENE (Roche) in 6-well plates. Cells were then grown in DMEM medium containing 0.5 ⁇ g/ml puromycin permanently. Western blot
  • RNA from cells or tumors was prepared in 2-4 ml of Trizol reagent (Invitrogen, Amsterdam, The Netherlands) according to Chomczynski and Sacchi (20). A total of 1 ⁇ g RNA was reverse transcribed using Superscript II reverse transcriptase (Invitrogen) following manufacturer's instructions and resuspended in 12 ⁇ final volume.
  • PCR polymerase chain reaction
  • 20 ⁇ reactions containing 0.5 ⁇ sense and antisense primers (Eurogentec, Angers, France) ; 0.6 ⁇ dNTP (20 mM) ; 2 ⁇ 1 of Taq polymerase (New England Biolabs, Herts, UK) at 5000 u/ ⁇ of commercial buffer for a total of 22 or 28 cycles consisting of 94°C for 40s, 57°C for 40s and 73°C for 60s for actin, S 100A10 and calpain 2.
  • the Hybridization temperature for annexin A2 was of 64°C.
  • Ten microliters amplification products were analyzed by electrophoresis in ethidium bromide- stained agarose gels. Primer sequences are available upon request.
  • HT-29R cells were forward transfected with control siRNA or a pool of 3 siRNAs (Invitrogen, stealth RNAi) directed against either S 100A10 (HSS 143791,143792, 143793), annexin A2 (HSS 179172, 179173, 179174) or calpain 2 (HSS 101347, 101348, 188705) using the lipofectamine RNAimax (Invitrogen) protocol.
  • RNAs were extracted using the Rneasy kit Mini (Qiagen) and quantified by nanodrop spectrophotometry. RNA quality was evaluated using the Agilent Bioanalyser 2100 and Lab-on -Chip Nano 6000 chip (ratio of the 28S / 18S RNA > 1.5).
  • Microarrays dedicated for study of cancer 2728 genes linked to proliferation, cell death, cell signalling, invasion, migration and inflammation) were printed using a selection of oligonucleotides from the human Reseau National des Genopoles/Medical Research Council oligonucleotides collection (21). The list of the 2728 probes spotted on the microarray is available on http://www. microarray.
  • RNA were labeled and hybridized as previously described (22).
  • Experimental data and associated microarray designs have been deposited in the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) (http://www.ncbi.nlm.nih.gov/geo/) under series GSE23433 and platform record GPL4719.
  • NCBI National Center for Biotechnology Information
  • GEO Gene Expression Omnibus
  • Ontologies attached to each modulated gene were then used to classified theme according to main biological themes.
  • Ingenuity software http://www.ingeniiity.com/.
  • the data were analyzed for themes using the gene ontology cellular compartment, molecular function, and biological process provided by Ingenuity.
  • Ingenuity software and Mediante 24
  • an information system containing diverse information about the probes set and the data sets http://www.microarray.fr
  • mice Fifty mice were inoculated s.c. with HT-29 human colon tumor cells. Treatment started when the mean tumor volume was 150 + 38 mm 3 ( Figure 1). After 7 weeks, no significant differences in tumor size were observed between control mice and mice treated with AS602868. These mice had to be euthanized for ethical reasons. After 10 weeks of treatment (first course), CPT-11 delayed tumor development (p ⁇ 0.0058) and as previously shown (10), the addition of AS602868 significantly improved CPT-11 antitumor effect (p ⁇ 0.0356). At that time, 10 out of 20 mice of the group which was treated with the bitherapy were euthanized, their tumors harvested, frozen and named "sensitive tumors". For the 10 remaining mice, the treatment was interrupted for 2 weeks.
  • mice received a second course of bitherapy for 10 weeks. From week 11 till 15, the tumors grew very quickly. From week 15 till 19, the bitherapy was efficient and tumor growth decreased. However, from week 19 till 22 (end of the second course of bitherapy), tumors escaped the treatment ( Figure 1). At the 22 nd week, the 10 mice were killed, their tumors removed and named "resistant tumors". From one of those tumors, a cell line was established, cultured and named HT-29R for resistant HT- 29 cells. The carcino-embryonic antigen (CEA) was similarly present on HT-29R and the HT-29 parental cell line showing the absence of contaminating cells (not shown).
  • CEA carcino-embryonic antigen
  • Figure 2A shows that in vitro HT-29R cells were not resistant to 3 ⁇ of the NF-KB inhibitor AS602868 but rather appeared slightly more sensitive. By contrast, HT-29R cells were significantly (p ⁇ 0.001) less sensitive to the cytotoxic effect of 10 nM CPT- l l/SN-38 associated or not to AS602868. These results have been confirmed in dose- response experiments using concentrations of AS602868 ranging from 0.3 to 10 ⁇ and from 3 to 100 nM for SN-38 (supplemental figure SI). The resistance of HT-29R cells to CPT-1 l/SN-38 appeared rather specific since HT-29R cells were not resistant to the cytotoxic effect of etoposide or 5-Fluorouracil (Figure 2B).
  • Unstimulated HT-29 cells displayed a weak constitutive activation of NF- ⁇ (Figure 3A, lane 1) that could be decreased by AS602868 (3 ⁇ , lane 2).
  • SN-38 (10 nM) induced a strong NF- ⁇ activation (1.8 fold, lane 3) that was affected by AS602868 (lane 4).
  • Similar results were observed in HT-29R cells except that the cells displayed a higher (1.8 x fold) constitutive NF- ⁇ activation (lane compared to lane 1) than in HT-29 cells.
  • a very high level of activated NF- ⁇ was also observed ex-vivo (Figure 3B) in 2 HT-29 resistant tumors compared to 2 HT-29 sensitive tumors.
  • HT-29R and RSN cells were stably transfected with a plasmid coding a super-repressor form of ⁇ - ⁇ which resulted in the down regulation of NF- ⁇ activation (Figure 4C).
  • inhibition of NF- ⁇ abrogated nearly completely the resistance of HT-29R cells and partially that of RSN cells to SN-38.
  • HT-29 resistant cells and tumors overexpress the potential NF- ⁇ inducers S100A10, Annexin A2 and Calpain 2
  • Figure 6B showed that the knockdown of SIOOAIO or annexin A2 by siRNA transfection for 3 days, had no effect on NF- ⁇ activation nor had that of S IOOAIO and annexin A2 (not shown).
  • knockdown of calpain 2 resulted in a sharp decrease in NF-KB activation in HT-29R and RSN resistant cells.
  • calpain 2 has been demonstrated to induce NF- ⁇ activation via the degradation of ⁇ - ⁇ (26, 28), Western blotting experiments studying ⁇ - ⁇ expression were performed.
  • HT-29R cells were then incubated or not with control or siCalpain 2 and the effect of increasing concentrations of CPT-l l/SN-38 on their viability was measured (Figure 6D). Calpain 2 knockown was able to significantly resensitize HT-29R cells to CPT-l l/SN-38 at all concentrations tested though calpain 2 knockdown could not completely restore the sensitivity of HT-29R to the level of the HT-29 parental cell line. Similar results were obtained in the resistant RSN cell line.
  • silencing S IOOAIO and/or Annexin A2 using siS lOOAlO and/or siAnnexin A2 had no effect on the sensitivity of HT-29R to CPT-l l/SN-38 (supplemental figure S6).
  • Vanhoefer U., Harstrick, A., Achterrath, W., Cao, S., Seeber, S., and Rustum, Y. M.
  • topoisomerase I messenger RNA, protein, and catalytic activity in human tumors demonstration of tumor-type specificity and implications for cancer chemotherapy. Cancer Res, 54: 539-546, 1994.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Cell Biology (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Food Science & Technology (AREA)
  • Biotechnology (AREA)
  • Hospice & Palliative Care (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Organic Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Zoology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne un inhibiteur de calpaïne pour utilisation pour retarder ou inhiber la résistance secondaire à un agent anticancéreux chez un patient souffrant de cancer colorectal. L'invention concerne en outre un procédé pour surveiller la réponse à un traitement d'un patient souffrant d'un cancer choisi parmi le cancer colorectal, le cancer gastrique, le cancer de l'œsophage, le cancer du poumon à grandes cellules et à petites cellules, la leucémie, des lymphomes, des gliomes malins du système nerveux central et des carcinomes ovariens, et de préférence le cancer colorectal, comprenant : a. le traitement dudit patient avec un agent anticancéreux pendant une durée d'au moins 5 mois ; puis b. la mesure du niveau d'expression d'au moins un gène choisi dans le groupe constitué des gènes de la calpaïne, l'annexine A2 et S100A10, dans des cellules cancéreuses dudit patient.
PCT/EP2012/052345 2011-02-14 2012-02-10 Procédés pour surveiller la réponse au traitement et pour traiter le cancer colorectal Ceased WO2012110425A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP11305153 2011-02-14
EP11305153.6 2011-02-14

Publications (1)

Publication Number Publication Date
WO2012110425A1 true WO2012110425A1 (fr) 2012-08-23

Family

ID=43770645

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2012/052345 Ceased WO2012110425A1 (fr) 2011-02-14 2012-02-10 Procédés pour surveiller la réponse au traitement et pour traiter le cancer colorectal

Country Status (1)

Country Link
WO (1) WO2012110425A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015066212A3 (fr) * 2013-10-31 2015-12-30 University Of Hawaii Inhibiteurs de la calpaïne destinés au traitement de la maladie inflammatoire chronique de l'intestin et du cancer colorectal

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4946778A (en) 1987-09-21 1990-08-07 Genex Corporation Single polypeptide chain binding molecules
WO2004010937A2 (fr) * 2002-07-26 2004-02-05 Advanced Research & Technology Institute At Indiana University Methode de traitement du cancer
WO2005066371A2 (fr) * 2003-12-31 2005-07-21 The Penn State Research Foundation Procedes permettant de prevoir et de surmonter la resistance a la chimiotherapie dans le cancer de l'ovaire et de prevoir l'apparition du cancer du colon
EP2241334A1 (fr) * 2008-01-31 2010-10-20 Keio University Procédé de détermination de la sensibilité à un agent anticancéreux

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4946778A (en) 1987-09-21 1990-08-07 Genex Corporation Single polypeptide chain binding molecules
WO2004010937A2 (fr) * 2002-07-26 2004-02-05 Advanced Research & Technology Institute At Indiana University Methode de traitement du cancer
WO2005066371A2 (fr) * 2003-12-31 2005-07-21 The Penn State Research Foundation Procedes permettant de prevoir et de surmonter la resistance a la chimiotherapie dans le cancer de l'ovaire et de prevoir l'apparition du cancer du colon
EP2241334A1 (fr) * 2008-01-31 2010-10-20 Keio University Procédé de détermination de la sensibilité à un agent anticancéreux

Non-Patent Citations (32)

* Cited by examiner, † Cited by third party
Title
BRZOSKA, K.; SZUMIEL, I.: "Signalling loops and linear pathways: NF-kappaB activation in response to genotoxic stress", MUTAGENESIS, vol. 24, 2009, pages 1 - 8
CAMP, E. R.; LI, J.; MINNICH, D. J.; BRANK, A.; MOLDAWER, L. L.; MACKAY, S. L.; HOCHWALD, S. N.: "Inducible nuclear factor-kappaB activation contributes to chemotherapy resistance in gastric cancer", J AM COLL SURG, vol. 199, 2004, pages 249 - 258
CHOMCZYNSKI, P.; SACCHI, N.: "Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction", ANAL BIOCHEM, vol. 162, 1987, pages 156 - 159, XP025650376, DOI: doi:10.1016/0003-2697(87)90021-2
CHU, X. Y.; SUZUKI, H.; UEDA, K.; KATO, Y.; AKIYAMA, S.; SUGIYAMA, Y.: "Active efflux of CPT-11 and its metabolites in human KB-derived cell lines", J PHARMACOL EXP THER, vol. 288, 1999, pages 735 - 741
CUSACK JAMES C JR ET AL: "Inducible chemoresistance to 7-ethyl-10-(4-(1-piperidino)-1-piperidin o)-carbonyloxycamptothecin (CPT-11) in colorectal cancer cells and a xenograft model is overcome by inhibition of nuclear factor-kappaB activation", CANCER RESEARCH, vol. 60, no. 9, 1 May 2000 (2000-05-01), pages 2323 - 2330, XP002632456, ISSN: 0008-5472 *
DEL RIO, M.; MOLINA, F.; BASCOUL-MOLLEVI, C.; COPOIS, V.; BIBEAU, F.; CHALBOS, P.; BAREIL, C.; KRAMAR, A.; SALVETAT, N.; FRASLON,: "Gene expression signature in advanced colorectal cancer patients select drugs and response for the use of leucovorin, fluorouracil, and irinotecan", J CLIN ONCOL, vol. 25, 2007, pages 773 - 780, XP002663356, DOI: doi:10.1200/JCO.2006.07.4187
DIGNAM, J. D.; LEBOVITZ, R. M.; ROEDER, R. G.: "Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei", NUCLEIC ACIDS RES, vol. 11, 1983, pages 1475 - 1489
FRELIN, C.; IMBERT, V.; GRIESSINGER, E.; PEYRON, A. C.; ROCHET, N.; PHILIP, P.; DAGEVILLE, C.; SIRVENT, A.; HUMMELSBERGER, M.; BER: "Targeting NF-kappaB activation via pharmacologic inhibition of IKK2-induced apoptosis of human acute myeloid leukemia cells", BLOOD, vol. 105, 2005, pages 804 - 811
HAN, Y.; WEINMAN, S.; BOLDOGH, I.; WALKER, R. K.; BRASIER, A. R.: "Tumor necrosis factor-alpha-inducible IkappaBalpha proteolysis mediated by cytosolic m-calpain. A mechanism parallel to the ubiquitin-proteasome pathway for nuclear factor-kappab activation", J BIOL CHEM, vol. 274, 1999, pages 787 - 794
HUSAIN, 1.; MOHLER, J. L.; SEIGLER, H. F.; BESTERMAN, J. M.: "Elevation of topoisomerase I messenger RNA, protein, and catalytic activity in human tumors: demonstration of tumor-type specificity and implications for cancer chemotherapy", CANCER RES, vol. 54, 1994, pages 539 - 546, XP055057626
JANSSENS, S.; TSCHOPP, J.: "Signals from within: the DNA-damage-induced NF-kappaB response", CELL DEATH DIFFER, vol. 13, 2006, pages 773 - 784
JEMAL, A.; SIEGEL, R.; WARD, E.; MURRAY, T.; XU, J.; THUN, M. J.: "Cancer statistics", CA CANCER J CLIN, vol. 57, 2007, pages 43 - 66
KARIN, M.: "How NF-kappaB is activated: the role of the IkappaB kinase (IKK) complex", ONCOGENE, vol. 18, 1999, pages 6867 - 6874
LAGADEC P ET AL: "Pharmacological targeting of NF-kappaB potentiates the effect of the topoisomerase inhibitor CPT-11 on colon cancer cells.", BRITISH JOURNAL OF CANCER 29 JAN 2008 LNKD- PUBMED:18182997, vol. 98, no. 2, 29 January 2008 (2008-01-29), pages 335 - 344, XP002632455, ISSN: 0007-0920 *
LAGADEC, P.; GRIESSINGER, E.; NAWROT, M. P.; FENOUILLE, N.; COLOSETTI, P.; IMBERT, V.; MARI, M.; HOFMAN, P.; CZERUCKA, D.; ROUSSEA: "Pharmacological targeting of NF-kappaB potentiates the effect of the topoisomerase inhibitor CPT-11 on colon cancer cells", BR J CANCER, vol. 98, 2008, pages 335 - 344, XP002632455, DOI: doi:10.1038/SJ.BJC.6604082
LE BRIGAND, K.; BARBRY, P.: "Mediante: a web-based microarray data manager", BIOINFORMATICS, vol. 23, 2007, pages 1304 - 1306
LE BRIGAND, K.; RUSSELL, R.; MOREILHON, C.; ROUILLARD, J. M.; JOST, B.; AMIOT, F.; MAGNONE, V.; BOLE-FEYSOT, C.; ROSTAGNO, P.; VIR: "An open-access long oligonucleotide microarray resource for analysis of the human and mouse transcriptomes", NUCLEIC ACIDS RES, vol. 34, 2006, pages E87
LI, Q.; LAUMONNIER, Y.; SYROVETS, T.; SIMMET, T.: "Plasmin triggers cytokine induction in human monocyte-derived macrophages", ARTERIOSCLER THROMB VASE BIOL, vol. 27, 2007, pages 1383 - 1389
LIU, L. F.; DESAI, S. D.; LI, T. K.; MAO, Y.; SUN, M.; SIM, S. P.: "Mechanism of action of camptothecin", ANN N Y ACAD SCI, vol. 922, 2000, pages 1 - 10
MALIEPAARD, M.; VAN GASTELEN, M. A.; TOHGO, A.; HAUSHEER, F. H.; VAN WAARDENBURG, R. C.; DE JONG, L. A.; PLUIM, D.; BEIJNEN, J. H.: "Circumvention of breast cancer resistance protein (BCRP)-mediated resistance to camptothecins in vitro using non-substrate drugs or the BCRP inhibitor GF120918", CLIN CANCER RES, vol. 7, 2001, pages 935 - 941
MOREILHON, C.; GRAS, D.; HOLOGNE, C.; BAJOLET, 0.; COTTREZ, F.; MAGNONE, V.; MERTEN, M.; GROUX, H.; PUCHELLE, E.; BARBRY, P.: "Live Staphylococcus aureus and bacterial soluble factors induce different transcriptional responses in human airway cells", PHYSIOL GENOMICS, vol. 20, 2005, pages 244 - 255
PAHL, H. L.: "Activators and target genes of Rel/NF-kappaB transcription factors", ONCOGENE, vol. 18, 1999, pages 6853 - 6866
PAQUET-DURAND F ET AL., JOURNAL OF NEUROCHEMISTRY, vol. 115, 2010, pages 930 - 940
SRIRANGAM ANJAIAH ET AL: "Ritonavir inhibition of m-calpain blocks colon cancer cell proliferation and synergizes with celecoxib", PROCEEDINGS OF THE ANNUAL MEETING OF THE AMERICAN ASSOCIATION FOR CANCER RESEARCH, AMERICAN ASSOCIATION FOR CANCER RESEARCH, US, vol. 44, 1 July 2003 (2003-07-01), pages 1113 - 1114, XP001525470, ISSN: 0197-016X *
VAN DE LOOSDRECHT, A. A.; BEELEN, R. H.; OSSENKOPPELE, G. J.; BROEKHOVEN, M. G.; LANGENHUIJSEN, M. M.: "A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia", J IMMUNOL METHODS, vol. 174, 1994, pages 311 - 320, XP023987425, DOI: doi:10.1016/0022-1759(94)90034-5
VANHOEFER, U.; HARSTRICK, A.; ACHTERRATH, W.; CAO, S.; SEEBER, S.; RUSTUM, Y. M.: "Irinotecan in the treatment of colorectal cancer: clinical overview", J CLIN ONCOL, vol. 19, 2001, pages 1501 - 1518, XP009052899
WEAVER, K. D.; YEYEODU, S.; CUSACK, J. C., JR.; BALDWIN, A. S., JR.; EWEND, M. G.: "Potentiation of chemotherapeutic agents following antagonism of nuclear factor kappa B in human gliomas", J NEUROONCOL, vol. 61, 2003, pages 187 - 196
WETHINGTON, S. L.; WRIGHT, J. D.; HERZOG, T. J.: "Key role of topoisomerase I inhibitors in the treatment of recurrent and refractory epithelial ovarian carcinoma", EXPERT REV ANTICANCER THER, vol. 8, 2008, pages 819 - 831
WETTENHALL, J. M.; SMYTH, G. K.: "limmaGUI: a graphical user interface for linear modeling of microarray data", BIOINFORMATICS, vol. 20, 2004, pages 3705 - 3706
XU, Y.; VILLALONA-CALERO, M. A.: "Irinotecan: mechanisms of tumor resistance and novel strategies for modulating its activity", ANN ONCOL, vol. 13, 2002, pages 1841 - 1851, XP002988009, DOI: doi:10.1093/annonc/mdf337
ZHU HONGBO ET AL: "Overcoming acquired resistance to TRAIL by chemotherapeutic agents and calpain inhibitor I through distinct mechanisms.", MOLECULAR THERAPY : THE JOURNAL OF THE AMERICAN SOCIETY OF GENE THERAPY MAY 2004 LNKD- PUBMED:15120327, vol. 9, no. 5, May 2004 (2004-05-01), pages 666 - 673, XP002632454, ISSN: 1525-0016 *
ZOBIACK, N.; GERKE, V.; RESCHER, U.: "Complex formation and submembranous localization of annexin 2 and S100A10 in live HepG2 cells", FEBS LETT, vol. 500, 2001, pages 137 - 140, XP004251394, DOI: doi:10.1016/S0014-5793(01)02604-7

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015066212A3 (fr) * 2013-10-31 2015-12-30 University Of Hawaii Inhibiteurs de la calpaïne destinés au traitement de la maladie inflammatoire chronique de l'intestin et du cancer colorectal

Similar Documents

Publication Publication Date Title
JP2021168674A (ja) 血中循環腫瘍細胞に関する方法およびアッセイ
US20120004289A1 (en) Annexin a11 and associated genes as biomarkers for cancer
US20100292303A1 (en) Gene expression profile for predicting ovarian cancer patient survival
US8772243B2 (en) Methods for predicting the response to anti-cancer treatment with an agonist of TLR7 or an agonist of TLR8
US20130224192A1 (en) Method for the prognosis of the progression of cancer
WO2010040083A2 (fr) Prédicteurs de chimiorésistance par expression génique
EP3779447B1 (fr) Procédé d'activation de la réponse antitumorale des cellules cd8+t d'un patient atteint d'un cancer
JP6587288B2 (ja) がん幹細胞の同定ならびに診断および処置のためのその使用
Wilhelm et al. Troy is expressed in human stomach mucosa and a novel putative prognostic marker of intestinal type gastric cancer
KR20170141951A (ko) Nckap1을 유효성분으로 포함하는 대장암 진단 또는 대장암 전이 예후 예측용 바이오마커 조성물
Zhang et al. Overexpressed CES2 has prognostic value in CRC and knockdown CES2 reverses L-OHP-resistance in CRC cells by inhibition of the PI3K signaling pathway
Yin et al. Ubiquitin-specific peptidase 15 regulates the TFAP4/PCGF1 axis facilitating liver metastasis of colorectal cancer and cell stemness
KR102141997B1 (ko) Pmvk를 유효성분으로 포함하는 방사선 저항성 암 진단용 또는 방사선 치료 예후 예측용 바이오마커 조성물
WO2012110425A1 (fr) Procédés pour surveiller la réponse au traitement et pour traiter le cancer colorectal
WO2017069710A1 (fr) Composition pour moduler irak1
WO2018232152A1 (fr) Procédés de traitement de cancers pharmacorésistants
US20110070245A1 (en) Imp-1 oncogene as a therapeutic target and prognostic indicator for lung cancer
KR20170129620A (ko) Lrp-1을 유효성분으로 포함하는 방사선 저항성 암 진단용 또는 방사선 치료 예후 예측용 바이오마커 조성물
Wang et al. AKIP1 promotes tumor progression by cancer-related pathways and predicts prognosis in tongue squamous cell carcinoma
US20130323231A1 (en) Treatment and prognosis of solid tumour cancers
EP2610620A1 (fr) Co-traitement d'inhibiteur de l'histone désacétylase 10 dans le cancer
WO2016026933A1 (fr) Traitement adjuvant ou néoadjuvant de sensibilisation de cellules souches cancéreuses à une chimiothérapie
US20240401146A1 (en) Methods for predicting and improving the efficacy of mcl-1 inhibitor therapy
WO2015132333A1 (fr) Procédés pour prédire la réponse au traitement et pour traiter le cancer
WO2024254217A2 (fr) Nouveaux biomarqueurs pour thérapies antitumorales et prdx6 en tant que cible thérapeutique pour le cancer

Legal Events

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

Ref document number: 12703318

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 12703318

Country of ref document: EP

Kind code of ref document: A1