[go: up one dir, main page]

WO2011110546A2 - Composition pharmaceutique comprenant un inhibiteur de shp-2 - Google Patents

Composition pharmaceutique comprenant un inhibiteur de shp-2 Download PDF

Info

Publication number
WO2011110546A2
WO2011110546A2 PCT/EP2011/053446 EP2011053446W WO2011110546A2 WO 2011110546 A2 WO2011110546 A2 WO 2011110546A2 EP 2011053446 W EP2011053446 W EP 2011053446W WO 2011110546 A2 WO2011110546 A2 WO 2011110546A2
Authority
WO
WIPO (PCT)
Prior art keywords
shp
pharmaceutical composition
inhibitor
hbd3
egfr
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/EP2011/053446
Other languages
English (en)
Other versions
WO2011110546A3 (fr
Inventor
Bianca Bauer
Thomas F. Meyer
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.)
Max Planck Gesellschaft zur Foerderung der Wissenschaften
Original Assignee
Max Planck Gesellschaft zur Foerderung der Wissenschaften
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 Max Planck Gesellschaft zur Foerderung der Wissenschaften filed Critical Max Planck Gesellschaft zur Foerderung der Wissenschaften
Publication of WO2011110546A2 publication Critical patent/WO2011110546A2/fr
Publication of WO2011110546A3 publication Critical patent/WO2011110546A3/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • 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
    • A61K31/13—Amines
    • A61K31/15—Oximes (>C=N—O—); Hydrazines (>N—N<); Hydrazones (>N—N=) ; Imines (C—N=C)
    • 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
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • 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
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/18—Sulfonamides
    • 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
    • A61K31/28—Compounds containing heavy metals
    • A61K31/285—Arsenic compounds
    • 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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/351—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom not condensed with another ring
    • 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
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/4162—1,2-Diazoles condensed with heterocyclic ring systems
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/429—Thiazoles condensed with heterocyclic ring systems
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/4439—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. omeprazole
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • 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
    • A61K31/4985—Pyrazines or piperazines ortho- or peri-condensed with heterocyclic ring systems
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
    • 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
    • A61K31/555—Heterocyclic compounds containing heavy metals, e.g. hemin, hematin, melarsoprol
    • 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
    • A61K31/66—Phosphorus compounds
    • A61K31/662—Phosphorus acids or esters thereof having P—C bonds, e.g. foscarnet, trichlorfon
    • A61K31/663—Compounds having two or more phosphorus acid groups or esters thereof, e.g. clodronic acid, pamidronic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material
    • 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
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G01N33/5041—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects involving analysis of members of signalling pathways
    • 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
    • C12N2799/00—Uses of viruses
    • C12N2799/02—Uses of viruses as vector
    • C12N2799/021—Uses of viruses as vector for the expression of a heterologous nucleic acid
    • C12N2799/027—Uses of viruses as vector for the expression of a heterologous nucleic acid where the vector is derived from a retrovirus
    • 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
    • C12N2799/00—Uses of viruses
    • C12N2799/02—Uses of viruses as vector
    • C12N2799/06—Uses of viruses as vector in vitro
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/205—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Campylobacter (G)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/71—Assays involving receptors, cell surface antigens or cell surface determinants for growth factors; for growth regulators

Definitions

  • composition comprising an inhibitor of SHP-2
  • Helicobacter pylori inhibits EGF receptor to shut down expression of antimicrobial defensin hBD3,
  • Defensins are short peptides that are crucial for the first line of innate immune responses in mammals.
  • Several defensins are known to be induced after infection with the human gastric pathogen Helicobacter pylori, including the human ⁇ -defensin hBD3.
  • H. pylori escapes from the highly potent antimicrobial effect of hBD3.
  • H. pylori disables hBD3 expression by directly inhibiting transactivation of the epidermal growth factor receptor (EGFR).
  • EGFR blockage is mediated by the translocated bacterial effector protein CagA and the tyrosine phosphatase SHP-2.
  • Our study provides a tantalizing mechanistic explanation of how the pathogenicity factor CagA enables H. pylori to persist in the human stomach over decades.
  • the highly adapted human gastric pathogen H. pylori colonizes at least half of the worlds population and elicits a tremendous socio-economic burden because of its etiological association with peptic ulcer disease and gastric cancer.
  • Infections with strains, expressing a functional type IV secretion system (TFSS) result in the translocation of the CagA effector protein into host epithelial cells, thereby enhancing the pathology of infection (Hatakeyama, 2004).
  • TFSS functional type IV secretion system
  • CagA targets the oncogenic tyrosine phosphatase SHP-2 to manipulate host cell signaling (Higashi et al., 2002).
  • defensins are potent antimicrobial peptides, referred to as defensins, during the initial stages of infection with different bacteria, including H. pylori (Lee and Josenhans, 2005;Menendez and Brett, 2007).
  • Defensins are short peptides and comprise key elements of the initial innate immune response against infection, exerting their antimicrobial impact via charge-mediated binding to bacterial membranes. Following membrane integration, defensins form pores resulting in bacterial lysis (Selsted and Ouellette, 2005). Recent work has shown that Nodl-dependent NF- ⁇ signaling mediates H.
  • HBD2 human ⁇ -defensin 2
  • EGFR epidermal growth factor receptor
  • H. pylori stimulates synthesis and secretion of antimicrobial hBD3 in gastric epithelial cells.
  • hBD3 has a known potent antimicrobial impact on a variety bacterial pathogens (Moranta et al., 2010), we wanted to assess its role during H. pylori infection in greater detail.
  • PI human gastric epithelial cells
  • Fig. 1A Infection of human gastric epithelial cells (AGS) with all three strains increased expression levels of hBD3 after 4 h (Fig. 1A).
  • AGS human gastric epithelial cells
  • Fig. 1A the viability of all tested H. pylori strains was drastically reduced in comparison to untreated controls, following incubation for 1 h with different concentrations of hBD3 peptides (Fig. IB).
  • hBD3 To delineate the host cell signaling pathways inducing expression of hBD3 ; we treated H. pylori infected cells with chemical inhibitors targeting EGFR and the MAP kinase MEK. Interestingly, hBD3 mRNA and protein expression levels were substantially reduced upon application of two different EGFR-specific inhibitors (AG1478, Gefitinib; Fig. 2A and fig. SI). Similarly, hBD3 protein expression was markedly diminished in infected cells treated with U0126, a common inhibitor of MEK (Fig. 2B). This data demonstrates a key role of the EGFR - MEK signaling cascade in the H. pylori dependent up-regulation of hBD3.
  • the EGFR is known to be activated by H. pylori during the early stages of infection (Keates et al. ( 2005;Wallasch et al., 2002), however, its activation status after prolonged infection periods in human cells remains unknown. Since initial hBD3 up-regulation is EGFR dependent (Fig. 2A and fig. SI), we examined EGFR transactivation during the course of an infection. Upon binding to its receptor, the EGFR-specific ligand EGF induces receptor dimerization and subsequent receptor autophosphorylation, referred to as transactivation, which can be monitored by immunoblotting. Therefore, we analyzed EGFR transactivation in non-infected and PI infected AGS cells after treatment with EGF for five minutes (Fig.
  • Plasmids encoding for either constitutive active EGFR (EGFR L858R) or EGFR wt (Uchida et al., 2007) were transiently expressed in non-infected and infected cells.
  • non- infected cells expressing EGFR L858R also produced hBD3, thus confirming the EGFR-dependent induction of hBD3 (Fig. 4D).
  • hBD3 mRNA levels were substantially higher in infected cells expressing EGFR L858R in comparison to non-infected cells. This finding is consistent with our previous observation that prolonged infection with H. pylori leads to enhanced surface exposition of EGFR (Bauer et al., 2009).
  • CagA dependent EGFR blockage is induced by the tyrosine phosphatase SHP-2.
  • CagA is known to activate the tyrosine phosphatase SHP-2 (Higashi et al., 2002). Further, SHP-2 is known to regulate EGFR signaling by mediating either direct EGFR dephosphorylation or dephosphorylation of downstream signaling molecules like GTPase activating proteins (Neel et al., 2003). We speculated blockage of EGFR could be the result of constant dephosphorylation; we therefore investigated the role of SHP-2 in the EGFR-mediated reduction of hBD3. Both chemical inhibition (ortho-vanadate) and siRNA-mediated depletion of SHP-2 resulted in the rescue of EGFR transactivation in infected cells (Fig. 5A and fig. S7).
  • SHP-2 mitigates the antimicrobial impact of hBD3 on bacterial viability.
  • H. pylori selectively disrupts SHP-2 dependent EGFR signaling to evade the antimicrobial impact of the human beta defensin hBD3
  • Endogenous antimicrobial peptides have been identified as key elements of innate host defence against infection.
  • defensins exhibit a variety of microbicidal activities against Gram-positive and -negative bacteria, mycobacteria, fungi and certain viruses.
  • the human beta defensin 3 (hBD3) was shown to have antimicrobial potential: it efficiently killed the human gastric pathogen Helicobacter pylori after short contact in vitro.
  • H. pylori type I strains persistently colonize the human stomach, thereby causing severe diseases like ulcer and gastric cancer. Whereas most bacteria are free swimming organisms in the gastric mucosa, a small percentage adheres to epithelial cells.
  • H. pylori pathogenic type I strains translocate the effector protein CagA into the host cell, which dramatically interferes with cellular signaling pathways like apoptosis and cytoskeletal rearrangements.
  • the mechanisms how H. pylori establishes a protective biological niche remain poorly understood. Since H. ry/on ' -mediated pathogenesis is a long-term process, this particular question is an important scientific issue. Therefore, we wanted to investigate if and how H. pylori escapes from the antimicrobial impact of hBD3 to enable persistent colonization. Intriguingly, we found that initially induced hBD3 expression vanishes over time in order to provide extracellular survival of H. pylori during longer infection.
  • H. pylori selectively blocks EGFR endocytosis via the nonreceptor kinase c-Abl and CagA. Cell Microbiol 11: 156-169.
  • pathogenicity island of Helicobacter pylori encodes type l-specific and disease-associated virulence factors. Proc Natl Acad Sci U S A 93: 14648-14653.
  • Type I H. pylori wild type strain PI, P12 and G27 are a clinical isolates, that has been described previously (Censini et al., 1996;Corthesy-Theulaz et al., 1996;Schmitt and Haas, 1994). Construction of the isogenic knock out mutants PlAcagA and PlAvirBll have been described elsewhere (Backert et al., 2001). Wildtype strains were cultivated on horse serum agar plates supplemented with vancomycin (10 ug/ml). Mutants were cultivated on selective chloramphenicol (4 ug/ml) agar plates. Bacteria were incubated 1 day before infection at 37°C in a humidified anaerobic atmosphere containing 5% C0 2 , 5% 0 2 and gasmixture of 9:1 N 2 /H 2 (Thermo Fischer).
  • lxlO 6 cells were infected with H. pylori for 24 h by using a MOI of 50.
  • Supernatant was diluted in BHI medium in a final volume of 1 ml (10 "1 , 10 '2 , 10 "3 and 10 "4 ) and 20 ⁇ of each dilution was plated on agar plates and incubated by 37°C (see bacterial cultivation). Cells with attached bacteria were washed 3 times with RPMI medium. 700 ⁇ 0.5% Saponin/BHI was added and incubated for 10 min by 37°C.
  • Lysed cells were homogenized thoroughly by pipetting and bacterial dilutions were prepared in a final volume of 1 ml (It) "1 , 10 "2 , 10 "3 and 10 “4 ). 20 ⁇ of each dilution was plated on agar plates (see bacterial cultivation) and incubated by 37°C. CFU (colony forming units) of attached (PE) and free swimming bacteria in the supernatant (SN) were calculated 3 days later.
  • It 1 ml
  • 1x10 s bacteria were incubated for 1 h by 37°C with or without indicated concentrations of hBD3. Bacteria were diluted subsequently in BHI medium in a final volume of 1 ml 10 "2 , 10 " 3 and 10 "4 ). 20 ⁇ of each bacterial suspension was plated on agar plates and incubated for 3 days (see bacterial cultivation). CFU (colony forming units) were counted and normalized to untreated bacterial samples.
  • the human gastric adenocarcinoma cell line AGS was grown in PMI 1640 medium containing 4 mM glutamine (Invitrogen) and 10% fetal calf serum (Gibco). Cells were incubated in a humidified atmosphere with 5% C0 2 . 2 days before infection cells were seeded in tissue culture plates. EGF and HB-EGF stimulations were performed after serum starvation for 20 h by adding 100 ng/ml EGF (Sigma) or 100 ng/ml HB-EGF (Sigma) for the indicated length of time.
  • Bacteria were collected from plates and resuspended in PBS. To avoid BHI components derived from the agar plates during infection, bacteria were washed once with PBS. After centrifugation for 3 min by 4000 g, bacteria were resuspended again in PBS. The amount of bacterial suspension required for indicated MOI (Multiplicity of infection) was calculated by measuring optical densities (OD 550 ) and by calculating respective volumes using a standard curve. Eucaryotic cells were infected under serum starved conditions and incubated at 37°C in a 5% C0 2 incubator for indicated time points. For long term infections (1 to 3 days) cells were grown in FCS containing medium to a confluency of 20% and infected with a MOI of 20 or 100. Infection of fallopian tubes was performed by adding 1 x 10 8 bacteria directly on the epithelial surface from tubular tissue.
  • AGS cells were transiently transfected with DNA or siRNA respectively.
  • Lipofectamine 2000 was used following the manufacturer's instructions (Invitrogen).
  • RNA transfections were performed by using RNAifect (Invitrogen).
  • DNA transfected cells were incubated at 37°C and 5% C0 2 24 h prior infection.
  • siRNA transfected cells were incubated under the same conditions for 24 h, subsequently splitted and further incubated for additional 48 h.
  • the plasmid encoding the constitutive active EGFR (EGFR L858R) and EGFR wildtype (EGFR WT) were a generous gift from Prof. Katsuyuki Kiura and Dr. Akiko Uchida (Uchida et al., 2007).
  • the plasmid encoding for SHP-2 was kindly provided by Dr. Paul Crocker (Avril et al., 2004).
  • SHP-2 (2r) AAGGUGGUUUCAUGGACAUCU; SHP-2 (1) GAAUAUGGCGUCAUGCGUGTT; SHP-2 (2) CCUUAACACGACUCGUAUAAA; SHP-2 (3) GCGUCAUGCGUGUUAGGAACG.
  • Controls were performed under the application of siRNA oligos targeting Luciferase (AACUUACGCUGAGUACUUCGA), which have no specific target in the genome.
  • shRNAs for SHP-2 constructs were stably transduced into AGS cells using a lentiviral based expression system (Wiznerowicz and Trono, 2003). Design of shRNAs was performed with the BLOCK-iTTM RNAi Designer from Invitrogen and cloned into the vector pLVTHM. 293T cells were transfected using calcium phosphate transfection together with the packaging vectors psPAX2 und pMD2G. Viruses were harvested from the supernatant and used for infection of HeLa cells in the presence of polybrene (5 ⁇ g/ml). GFP-positive cells were selected 7 days p.i. and sorted into 96-well plates using flow cytometry. Target sequences for SHP-2 constructs: SHP2_1: GCCTCATATGTTGAATCATCC; SHP2_4: G G G ATCAACTTTC ACAC ATAG;
  • Fallopian tubes were obtained from women undergoing operative procedures for tubal ligation or hysterectomy. Material was transported on ice within 2 hours after operation. Extra connective tissue was removed by dissection under continuous washing with PBS. Tubular lumen was identified and rinsed with PBS by using a syringe. Transversal sections (approx. 1-2 mm) were generated by using a scalpel. Tissue slices were transferred to small petri dishes in MEM Earl medium+10 % FCS and incubated at 37 °C prior to infection. 1 h post infection tissue was fixed by incubating samples 24 h in 4% PFA. Samples were stored in PBS at 4°C prior to paraffin embedding.
  • Paraffin embedding and immunofluorescence labelling was performed as described by (Robertson et al., 2008). Polyclonal anti Urease-B antibody was used to stain H. pylori, DraQ 5 (Alexis) was applied to stain nuclei and monoclonal anti-hBD3 antibody (Abnova) was used to visualize hBD3. All secondary antibodies were purchased from Jackson ImmunoResearch. Samples were analyzed by confocal laser scanning microscopy using a Leica TCS SP microscope.
  • Lysates were incubated with 1 ⁇ monoclonal anti c-Met antibody (Upstate 05-237) or anti SHP-2 (Santa Cruz) antibody by 4 °C overnight under subsequent addition of protein G-sepharose beads for 2 h by 4 °C. Precipitates were analyzed by SDS-PAGE and immunoblotting.
  • Immunoblot quantification of band intensities were performed from digital pictures which were processed using the software Photoshop CS3 (Adobe). Pictures were inverted and region of interest was defined. Histogram of selected region displayed median of intensity and region size. All bands were normalized to background regions and respective loading control band intensities.
  • hBD3 FW 5 " TCTGCCTTACCATTGGGTTC 3 ' ; hBD3 RW: 5 ' CACGCTGAGACTGGATGAAA 3 " ; GAPDH FW: 5 * GGTATCGTGGAAGGACTCATGAC 3 ' ; GAPDH RW: 5 ' ATG CC AGTG AG CTTCCCGTTCAG 3 '
  • Figure 1 Infection with H. pylori induce expression of antimicrobial hBD3 in gastric epithelial cells.
  • AGS cells were infected with PI, P12 and G27 for indicated time points using an MOI (multiplicity of infection) of 100.
  • MOI multiplicity of infection
  • Nl Infected and non-infected cells were lysed and analyzed for hBD3 expression by immunoblot. Actin was used as a loading control.
  • B Antimicrobial activity assay of hBD3 with three different H. pylori wildtype strains (PI, P12, G27). Bacteria (lxlO 5 ) were incubated for 1 h with indicated peptide concentrations. Colony forming units (CFU) were quantified three days later. Data are mean ⁇ SD of three independent experiments.
  • FIG. 2 hBD3 expression is mediated by EGFR and MAP kinases.
  • AGS cells were infected with PI using an MOI of 100. Before and during infection cells were treated with inhibitors against EGFR (AG1478, Gefitinib) or against B) MAP kinases (U0126).
  • A RT-PCR analysis of hBD3 mRNA expression in PI infected and non-infected (Nl) cells after 8 h of infection. Inhibitor added at 10 ⁇ ). DMSO was used as control in infected cells. Error bars indicate mean + SD of three independent experiments.
  • B hBD3 expression of infected and non-infected cells treated with (left panel) or without (right panel) U0126 was analyzed by immunoblot. Inhibitor used at 100 nM. Actin was used as a loading control.
  • Figure 3 Initially induced hBD3 expression decreases during prolonged infections.
  • A AGS cells were infected with PI using the indicated MOI. To compare hBD3 expression between prolonged and short infections, cells were infected for 2 h on different days (1,2 and 3 days). Final hBD3 expression was analyzed by immunoblotting. Actin was used as a loading control.
  • B, D RT-PCR analysis of hBD3 mRNA expression in infected and non-infected (Nl) AGS cells (MOI 100). Error bars indicate mean ⁇ SD of three independent experiments.
  • B Cells were infected with PI for indicated time points.
  • C Immunofluorescence staining of PI infected AGS cells 1 day (Id) and 2 days (2d) after infection.
  • hBD3 was stained by using an anti hBD3 antibody (red), bacteria are visualized with an anti CagA antibody (green).
  • Nuclei are stained with DraQ5 (blue). Scale bar represents 10 ⁇ . Analysis was performed by using confocal microscopy.
  • Figure 4 Reduction of hBD3 is caused by selective inhibition of EGFR in a CagA-dependent manner.
  • AGS cells were infected with PI using an MOI of 100. After infection cells were treated with 100 ng/ml EGF for 5 min.
  • A EGFR phosphorylation was measured by immunoblotting using an anti-phosphotyrosine antibody. Phosphorylation intensities were quantified and normalized to non-infected unstimulated cells. Error bars indicate mean ⁇ SD of three independent experiments.
  • B Immunoblot analysis of EGFR phosphorylation at specific tyrosines in infected (PI) and non-infected (Nl) cells. Antibodies against pY845, pY1045, pY992 and pY1068 were used. EGFR was used as a loading control.
  • FIG. 5 EGFR and hBD3 expression are inhibited via SHP-2.
  • AGS cells were infected with PI (MOI 100). Cells were analyzed by immunoblotting 24 h after infection. For EGFR phosphorylation analysis infected and non-infected (Nl) cells were stimulated with EGF (100 ng/ml) for 5 min. EGFR phosphorylation was analyzed by immunoblottin using an anti-phosphotyrosine antibody. Actin was used as a loading control. Error bars indicate mean + SD of three independent experiments. (A, B, C) AGS cells were transfected with siRNAs targeting SHP-2 or luciferase as control.
  • A, B Three days post transfection cells were infected and analyzed by immunoblot (A, B) or RT-PCR (C).
  • A Before lysis cells were treated with EGF. Phosphorylation intensities were quantified and normalized to non-infected unstimulated cells.
  • B Immunoblotting analysis of hBD3 protein expression. Knockdown efficiency of SHP-2 was monitored using an anti SHP-2 antibody.
  • C hBD3 mRNA quantification by RT-PCR of siRNA transfected cells.
  • D RT-PCR of hBD3 mRNA expression in infected and non-infected AGS and SHP-2 knockdown cells (SHP2_4_KD).
  • SHP-2 enables bacterial survival during prolonged infections.
  • A Immunofluorescence analysis of PI infected AGS and SHP-2 knock down cells (SHP2_4_KD). Cells were infected (MOI 100) and stained with an anti-hBD3 antibody (red) or anti-CagA antibody (blue) to visualize H. pylori. Knockdown cells express GFP (green) as a reporter construct for efficient lentiviral integration. Scale bar represents 10 ⁇ . Analysis was performed using confocal microscopy.
  • B CFU assay of free swimming (supernatant; SN) and attached (pellet; Pe) PI after infection of AGS and SHP-2 knockdown cells. Bacteria were plated after 24 h of infection.
  • C Systematic model of CagA and SHP-2-mediated blockage of hBD3 expression during later infection time points.
  • FIG. 10 hBD3 expression is mediated by EGFR.
  • AGS cells were infected with PI (MOI 100). Before and during infection cells were treated with the indicated inhibitor concentrations targeting EGFR (AG1478).
  • hBD3 expression of infected (PI) and non-infected (Nl) cells was analyzed by immunoblotting. Actin was used as a loading control.
  • Figure S2 Cellular viability of infected AGS cells.
  • AGS cells (lxlO 6 ) were infected with PI (MOI 100).
  • Cellular viability was measured 24 h after infection using a WST assay kit (company) and normalized to non-infected (Nl) cells. Error bars indicate mean ⁇ SD of three independent experiments. Cells treated for 5 min with TritonX-100 (TX) as a control.
  • TX TritonX-100
  • FIG. S3 EGFR inhibition is MOI dependent. AGS cells were infected with PI at the indicated MOI. Non- infected and infected cells were treated with EGF (100 ng/ml) 24 h after infection. Immunoblot analysis of EGFR phosphorylation was performed by using an anti phosphotyrosine antibody. Actin was used as a loading control.
  • Figure S4 EGFR Y1068 phosphorylation is inhibited during prolonged infections.
  • AGS cells were infected with PI (MOI 100).
  • Non-infected (Nl) and infected cells were stained with an antibody directed to EGFR pY1068 (red).
  • Scale bar represents 50 ⁇ . Immunofluorescence was performed by epifluorescence microscopy.
  • Figure S5 Blockage of EGFR transactivation is ligand-independent.
  • AGS cells were infected with PI (MOI 100).
  • Non-infected (Nl) and infected cells were stimulated at 24 h after infection with EGF or HB- EGF respectively for the indicated time points.
  • Immunoblotting analysis of EGFR phosphorylation was performed using an anti-phosphotyrosine antibody. Actin was used as a loading control.
  • Figure S6 H. pylori induced inhibition is selectively directed against EGFR.
  • c-Met was precipitated from infected and non-infected (Nl) cells 24 h p.i. Before lysis cells were treated with 100 ng/ml HGF. Phosphorylation of precipitated c-Met was analyzed by immunoblotting using an anti-phosphotyrosine antibody. Equal loading was confirmed with an anti c-Met antibody.
  • FIG. S7 EGFR inhibition is dependent on tyrosine phosphatases.
  • AGS cells were infected with PI (MOl 100). Non-infected (Nl) and infected cells were treated at 24 h after infection with different concentrations of activated orthovanadate. Cells were subsequently stimulated with EGF (100 ng/ml) for 5 min and cellular lysates were analyzed by immunoblot. Long and short exposure times are shown to visualize EGFR phosphorylation under different inhibitor conditions. Immunoblot analysis of EGFR phosphorylation was performed by using an anti-phosphotyrosine antibody. Actin was used as a loading control.
  • FIG. S8 SHP-2 is phosphorylated during H. pylori infection.
  • AGS cells were transfected with a plasmid encoding for SHP-2.
  • SHP-2 was precipitated using an anti SHP-2 antibody.
  • Precipitates of non-infected (Nl) and infected cells were analyzed by immunoblotting (lower panel).
  • SHP-2 phosphorylation was monitored using an anti-phosphotyrosine antibody. Equal loading was confirmed with an antibody directed to actin. Phosphorylation intensity was normalized to non-infected cells (upper panel). Error bars indicate mean ⁇ SD of three independent experiments.
  • Figure S9 SHP-2 knockdown in stable SHP2_4_KD cells.
  • AGS derived stable SHP-2 knockdown cells were lysed and analyzed by immunoblot.
  • AGS wildtype cells were used as control.
  • SHP-2 protein levels were monitored by using an anti SHP-2 antibody. Actin was used as a loading control.
  • NCI National Cancer Institute
  • NCI diversity set of compounds has led to the identification of 5 (NSC-117199) which inhibits the protein tyrosine phosphatase (PTP) Shp2 with an IC 50 of 47 ⁇ .
  • PTP protein tyrosine phosphatase
  • PH PSl is the first compound that specifically inhibits Shp2 over the closely related
  • PH PSl is as an active site-directed small molecule inhibitor of Shp2.
  • K i value of PH PSl for inhibition of Shp2 was 0.73 ( ⁇ 0.34) ⁇ , which is in agreement with the value, 0.7 ⁇
  • stibogluconate a drug used in treatment of leishmaniasis, is a potent inhibitor of PTPases Src homology PTPasel (SHP-1), SHP-2, and PTPIB but not the dual-specificity phosphatase mitogen- activated protein kinase phosphatase 1.
  • Each vial contains 200 mM Imidazole, 100 mM Sodium Fluoride, 115 mM Sodium Molybdate,
  • Non-competitive protein tyrosine phosphatase inhibitor enhances insulin signalling and insulin tolerance in ob/ob mice. Displays irreversible inhibition via catalysis of hydrogen peroxide- dependent oxidation of PTP.
  • p-Bromotetramisole is a well known inhibitor of alkaline phosphatases which was recently found to inhibit tyrosine phosphatase. This activity was identified by its ability to mimic the action of orthovanadate (a well known tyrosine phosphatase inhibitor) (1) in the potentiation of fluorouracil antiproliferative activity (2).
  • Ptp inhibitor IV ( j s(4-Trifluoromethylsulfonamidophenyl)-l,4-diisopropylbenzene);

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Wood Science & Technology (AREA)
  • Toxicology (AREA)
  • Analytical Chemistry (AREA)
  • Biotechnology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Hematology (AREA)
  • Microbiology (AREA)
  • Zoology (AREA)
  • Urology & Nephrology (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Communicable Diseases (AREA)
  • Food Science & Technology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biophysics (AREA)
  • Cell Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Oncology (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)

Abstract

La présente invention concerne une composition pharmaceutique comprenant au moins un inhibiteur de SHP-2, de façon facultative conjointement avec des véhicules, adjuvants, diluants et/ou additifs pharmaceutiquement acceptables. La présente invention concerne en outre des procédés de criblage pour l'identification d'un composé adapté en tant qu'inhibiteur de SHP-2.
PCT/EP2011/053446 2010-03-08 2011-03-08 Composition pharmaceutique comprenant un inhibiteur de shp-2 Ceased WO2011110546A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US31152910P 2010-03-08 2010-03-08
US61/311529 2010-03-08

Publications (2)

Publication Number Publication Date
WO2011110546A2 true WO2011110546A2 (fr) 2011-09-15
WO2011110546A3 WO2011110546A3 (fr) 2012-01-12

Family

ID=44486137

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2011/053446 Ceased WO2011110546A2 (fr) 2010-03-08 2011-03-08 Composition pharmaceutique comprenant un inhibiteur de shp-2

Country Status (1)

Country Link
WO (1) WO2011110546A2 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150352131A1 (en) * 2013-01-16 2015-12-10 Rhode Island Hospital Compositions and Methods for the Prevention and Treatment of Osteolysis and Osteoporosis
JP2023530089A (ja) * 2020-06-11 2023-07-13 シーエイチディーアイ ファウンデーション，インコーポレーテッド ハンチンチンタンパク質をイメージングするための複素環式化合物及びイメージング剤

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1728790A1 (fr) * 2005-06-01 2006-12-06 Max-Delbrück-Centrum Für Molekulare Medizin Inhibiteurs de Shp-2, compositions pharmaceutiques les contenant et leur utilisation pour le traitement de maladies induites par la Phosphatase
US8987474B2 (en) * 2006-04-07 2015-03-24 University Of South Florida Inhibition of Shp2/PTPN11 protein tyrosine phosphatase by NSC-87877, NSC-117199 and their analogs
WO2009135000A2 (fr) * 2008-04-30 2009-11-05 H. Lee Moffitt Cancer Center And Research Institute, Inc. Inhibition de la protéine tyrosine phosphatase shp2/ptpn11 par nsc-117199 et analogues

Non-Patent Citations (31)

* Cited by examiner, † Cited by third party
Title
ANDERSEN, L. P.; RASMUSSEN, L.: "Helicobacter pylori-coccoid forms and biofilm formation", FEMS IMMUNOL MED MICROBIOL, vol. 56, 2009, pages 112 - 115
BACKERT, S.; MOESE, S.; SELBACH, M.; BRINKMANN, V.; MEYER, T. F.: "Phosphorylation of tyrosine 972 of the Helicobacter pylori CagA protein is essential for induction of a scattering phenotype in gastric epithelial cells", MOL MICROBIOL42, 2001, pages 631 - 644
BAUER, B.; BARTFELD, S.; MEYER, T. F.: "H. pylori selectively blocks EGFR endocytosis via the non- receptor kinase c-Abl and CagA", CELL MICROBIAL, vol. 11, 2009, pages 156 - 169
BOUGHAN, P. K.; ARGENT, R. H.; BODY-MALAPEL, M.; PARK, J. H.; EWINGS, K. E.; BOWIE, A. G. ET AL.: "Nucleotide-binding oligomerization domain-1 and epidermal growth factor receptor: critical regulators of beta-defensins during Helicobacter pylori infection", J BIOL CHEM, vol. 281, 2006, pages 11637 - 11648
CENSINI, S.; LANGE, C.; XIANG, Z.; CRABTREE, J. E.; GHIARA, P.; BORODOVSKY, M. ET AL.: "cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors", PROC NATL ACAD SCI U S A, vol. 93, 1996, pages 14648 - 14653, XP002246958, DOI: doi:10.1073/pnas.93.25.14648
CHEN ET AL., J BIOL CHEM, 2001
CHEN ET AL., MOLECULAR PHARMACOLOGY, 2006
CORTHESY-THEULAZ; I., PORTA; N., PRINGAULT; E., RACINE; L., BOGDANOVA; A., KRAEHENBUHL ET AL.: "Adhesion of Helicobacter pylori to polarized T84 human intestinal cell monolayers is pH dependent", INFECT IMMUN, vol. 64, 1996, pages 3827 - 3832
GRUBMAN, A.; KAPARAKIS, M.; VIALA, J.; ALLISON, C.; BADEA, L.; KARRAR, A. ET AL.: "The innate immune molecule, NOD1, regulates direct killing of Helicobacter pylori by antimicrobial peptides", CELL MICROBIOL, 2009
HATAKEYAMA, M.: "Oncogenic mechanisms of the Helicobacter pylori CagA protein", NAT REV CANCER, vol. 4, 2004, pages 688 - 694
HELLMUTH ET AL., PROC NATL ACAD SCI, 2008
HIGASHI, H.; TSUTSUMI, R.; MUTO, S.; SUGIYAMA, T.; AZUMA, T.; ASAKA, M. ET AL.: "SHP-2 tyrosine phosphatase as an intracellular target of Helicobacter pylori CagA protein", SCIENCE, vol. 295, 2002, pages 683 - 686
HUBBARD, S. R.; MILLER, W. T.: "Receptor tyrosine kinases: mechanisms of activation and signaling", CURR OPIN CELL BIOL, vol. 19, 2007, pages 117 - 123, XP005931716, DOI: doi:10.1016/j.ceb.2007.02.010
KEATES, S.; KEATES, A. C.; NATH, S.; PEEK, R. M., JR.; KELLY, C. P.: "Transactivation of the epidermal growth factor receptor by cag+ Helicobacter pylori induces upregulation of the early growth response gene Egr-1 in gastric epithelial cells", GUT, vol. 54, 2005, pages 1363 - 1369
LAWRENCE ET AL., J MED CHEM, 2008
LEE, S. K.; JOSENHANS, C.: "Helicobacter pylori and the innate immune system", INT MED MICROBIOL, vol. 295, 2005, pages 325 - 334, XP005027076, DOI: doi:10.1016/j.ijmm.2005.06.004
MENENDEZ, A.; BRETT, F. B.: "Defensins in the immunology of bacterial infections", CURR OPIN IMMUNOL, vol. 19, 2007, pages 385 - 391, XP022208983, DOI: doi:10.1016/j.coi.2007.06.008
MORANTA, D.; REGUEIRO, V.; MARCH, C.; LLOBET, E.; MARGARETO, J.; LARRATE, E. ET AL.: "Klebsiella pneumoniae capsule polysaccharide impedes the expression of beta-defensins by airway epithelial cells", INFECT IMMUN, vol. 78, 2010, pages 1135 - 1146
NEEL, B. G.; GU, H.; PAO, L.: "The'Shp'ing news: SH2 domain-containing tyrosine phosphatases in cell signaling", TRENDS BIOCHEM SCI, vol. 28, 2003, pages 284 - 293, XP004433006, DOI: doi:10.1016/S0968-0004(03)00091-4
NÖREN-MÜLLER ET AL., PROC NATL ACAD SCI, 2006
PATHAK ET AL., THE J OF IMMUNOLOGY, 2001
ROBERTSON, D.; SAVAGE, K.; REIS-FILHO, J. S.; ISACKE, C. M.: "Multiple immunofluorescence labelling of formalin-fixed paraffin-embedded (FFPE) tissue", BMC CELL BIOL, vol. 9, 2008, pages 13, XP021032137
SCHMITT, W.; HAAS, R.: "Genetic analysis of the Helicobacter pylori vacuolating cytotoxin: structural similarities with the IgA protease type of exported protein", MOL MICROBIO/12, 1994, pages 307 - 319, XP000605827, DOI: doi:10.1111/j.1365-2958.1994.tb01019.x
SCHOENWAELDER ET AL., CURR BIOL, 2000
SELSTED, M. E.; OUELLETTE, A. J.: "Mammalian defensins in the antimicrobial immune response", NAT IMMUNOL, vol. 6, 2005, pages 551 - 557, XP002401789, DOI: doi:10.1038/ni1206
STRACK ET AL., BIOCHEMISTRY, 2002
SUN ET AL., JBC, 2003
UCHIDA, A.; HIRANO, S.; KITAO, H.; OGINO, A.; RAI, K.; TOYOOKA, S. ET AL.: "Activation of downstream epidermal growth factor receptor (EGFR) signaling provides gefitinib-resistance in cells carrying EGFR mutation", CANCER SCI, vol. 98, 2007, pages 357 - 363, XP055263322, DOI: doi:10.1111/j.1349-7006.2007.00387.x
WADA, A.; OGUSHI, K.; KIMURA, T.; HOJO, H.; MORI, N.; SUZUKI, S. ET AL.: "Helicobacter pylori-mediated transcriptional regulation of the human beta-defensin 2 gene requires NF-kappaB", CELL MICROBIOL3, 2001, pages 115 - 123
WALLASCH, C.; CRABTREE, J. E.; BEVEC, D.; ROBINSON, P. A.; WAGNER, H.; ULLRICH, A.: "Helicobacter pylori-stimulated EGF receptor transactivation requires metalloprotease cleavage of HB-EGF", BIOCHEM BIOPHYS RES COMMUN, vol. 295, 2002, pages 695 - 701, XP003003291, DOI: doi:10.1016/S0006-291X(02)00740-4
WIZNEROWICZ, M.; TRONO, D.: "Conditional suppression of cellular genes: lentivirus vector- mediated drug-inducible RNA interference", J VIROL, vol. 77, 2003, pages 8957 - 8961, XP002290538, DOI: doi:10.1128/JVI.77.16.8957-8951.2003

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20150352131A1 (en) * 2013-01-16 2015-12-10 Rhode Island Hospital Compositions and Methods for the Prevention and Treatment of Osteolysis and Osteoporosis
JP2023530089A (ja) * 2020-06-11 2023-07-13 シーエイチディーアイ ファウンデーション，インコーポレーテッド ハンチンチンタンパク質をイメージングするための複素環式化合物及びイメージング剤

Also Published As

Publication number Publication date
WO2011110546A3 (fr) 2012-01-12

Similar Documents

Publication Publication Date Title
Fajardo et al. The endoplasmic reticulum stress/unfolded protein response in gliomagenesis, tumor progression and as a therapeutic target in glioblastoma
Lodola et al. Store-operated Ca2+ entry is remodelled and controls in vitro angiogenesis in endothelial progenitor cells isolated from tumoral patients
Dirat et al. Inhibition of the GTPase Rac1 mediates the antimigratory effects of metformin in prostate cancer cells
Duan et al. Deficiency of ATP6V1H causes bone loss by inhibiting bone resorption and bone formation through the TGF-β1 pathway
Jamieson et al. Rac1 augments Wnt signaling by stimulating β-catenin–lymphoid enhancer factor-1 complex assembly independent of β-catenin nuclear import
US20160130663A1 (en) Method for predicting response to cancer treatment
CN105358173B (zh) 靶向癌症治疗的egfr-sglt1相互作用
Correa et al. The NLR-related protein NWD1 is associated with prostate cancer and modulates androgen receptor signaling
Wottawa et al. Knockdown of prolyl‐4‐hydroxylase domain 2 inhibits tumor growth of human breast cancer MDA‐MB‐231 cells by affecting TGF‐β1 processing
Dou et al. Sodium butyrate improves porcine host defense peptide expression and relieves the inflammatory response upon Toll-like receptor 2 activation and histone deacetylase inhibition in porcine kidney cells
Geng et al. 10-hydroxy-2-decenoic acid prevents osteoarthritis by targeting aspartyl β hydroxylase and inhibiting chondrocyte senescence in male mice preclinically
Yan et al. Molecular regulation and clinical significance of caveolin‐1 methylation in chronic lung diseases
Ye et al. NUMB maintains bone mass by promoting degradation of PTEN and GLI1 via ubiquitination in osteoblasts
Tong et al. Loss of Par3 promotes lung adenocarcinoma metastasis through 14-3-3ζ protein
Zhao et al. MicroRNA-939-5p directly targets IGF-1R to inhibit the aggressive phenotypes of osteosarcoma through deactivating the PI3K/Akt pathway
Zhang et al. SOCS6 Promotes Mitochondrial Fission and Cardiomyocyte Apoptosis and Is Negatively Regulated by Quaking‐Mediated miR‐19b
Yi et al. TMBIM6 deficiency leads to bone loss by accelerating osteoclastogenesis
US20200282015A1 (en) Targeted pharmacological therapeutics in uveal melanoma
Zhang et al. miR-802 inhibits the aggressive behaviors of non-small cell lung cancer cells by directly targeting FGFR1
Wang et al. Overexpression of myosin VI regulates gastric cancer cell progression
Sun et al. Cooperation of TRADD-and RIPK1-dependent cell death pathways in maintaining intestinal homeostasis
WO2011110546A2 (fr) Composition pharmaceutique comprenant un inhibiteur de shp-2
Akuma et al. Loss of VHL-mediated pRb regulation promotes clear cell renal cell carcinoma
Smith et al. c-JUN n-terminal kinase (JNK) signaling in autosomal dominant polycystic kidney disease
US20160362464A1 (en) Novel therapeutic targets for cancer progression

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: 11729927

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11729927

Country of ref document: EP

Kind code of ref document: A2