WO2002032429A2 - Use of inhibitors of progesterone receptor for treating cancer - Google Patents

Use of inhibitors of progesterone receptor for treating cancer Download PDF

Info

Publication number
WO2002032429A2
WO2002032429A2 PCT/EP2001/012004 EP0112004W WO0232429A2 WO 2002032429 A2 WO2002032429 A2 WO 2002032429A2 EP 0112004 W EP0112004 W EP 0112004W WO 0232429 A2 WO0232429 A2 WO 0232429A2
Authority
WO
WIPO (PCT)
Prior art keywords
use according
tumor
cells
progesterone receptor
growth
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/EP2001/012004
Other languages
French (fr)
Other versions
WO2002032429A3 (en
Inventor
Rosemarie Lichtner
Ulrike Fuhrmann
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.)
Bayer Pharma AG
Original Assignee
Schering AG
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
Priority claimed from DE10051609A external-priority patent/DE10051609A1/en
Priority to US10/399,318 priority Critical patent/US20040157811A1/en
Priority to HK05104537.7A priority patent/HK1071697B/en
Priority to HR20030386A priority patent/HRP20030386A2/en
Priority to JP2002535667A priority patent/JP4335525B2/en
Priority to EA200300468A priority patent/EA005945B1/en
Priority to SK475-2003A priority patent/SK4752003A3/en
Priority to KR10-2003-7005358A priority patent/KR20030044006A/en
Priority to MXPA03002953A priority patent/MXPA03002953A/en
Priority to HU0301461A priority patent/HUP0301461A3/en
Priority to DK01987667T priority patent/DK1414465T3/en
Priority to EEP200300156A priority patent/EE200300156A/en
Priority to EP01987667A priority patent/EP1414465B1/en
Priority to AU2002215958A priority patent/AU2002215958B2/en
Application filed by Schering AG filed Critical Schering AG
Priority to PL01366415A priority patent/PL366415A1/en
Priority to AU1595802A priority patent/AU1595802A/en
Priority to DE60123138T priority patent/DE60123138T2/en
Priority to UA2003054403A priority patent/UA76128C2/en
Priority to CA002423013A priority patent/CA2423013A1/en
Priority to BR0114677-7A priority patent/BR0114677A/en
Priority to IL15497401A priority patent/IL154974A0/en
Priority to SI200130665T priority patent/SI1414465T1/en
Publication of WO2002032429A2 publication Critical patent/WO2002032429A2/en
Priority to IL154974A priority patent/IL154974A/en
Priority to NO20031745A priority patent/NO20031745L/en
Priority to BG107745A priority patent/BG107745A/en
Anticipated expiration legal-status Critical
Publication of WO2002032429A3 publication Critical patent/WO2002032429A3/en
Priority to CY20061101786T priority patent/CY1106289T1/en
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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • 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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/565—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol
    • A61K31/567—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol substituted in position 17 alpha, e.g. mestranol, norethandrolone
    • 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
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/138—Aryloxyalkylamines, e.g. propranolol, tamoxifen, phenoxybenzamine
    • 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
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
    • A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to the use of progesterone receptor inhibitors for inhibiting the growth factor dependency of tumor cells .
  • Estradiol and progesterone are involved in the development of breast cancer. At the time of diagnosis, however, only about 1/3 of the tumors show a steroid hormone dependency. It is assumed that in the majority of steroid hormone-resistant tumors, the proliferation control for local -acting autocrine or paracrine peptidic growth factors is taken over. In this case, invasive tumors with extremely poor prognosis that are growth-factor- receptor-positive and steroid hormone-resistant result (Elledge et al . , Semin. Onkol . 19 (1992), 244-253).
  • the growth stimulation is limited to a single cycle, however, and is followed by a growth arrest at the Gl/S-transition of the second cycle (Groshong et al . (1997), supra: Musgrove et al . , Mol . Cell. Biol. 13 (1993), 3577-3587).
  • the cells are sensitive to the proliferative action of EGF.
  • progesterone enhances the action of EGF on T47D cells by ramping up EGFR, Erb2 and Erb3 and increases the tyrosine phosphorylation of signal molecules (Lange et al., J. Biol. Chem.
  • inhibitors of the progesterone receptor e.g., 17 ⁇ -fluoroalkyl steroids
  • a subject of this invention is thus the use of an inhibitor of the progesterone receptor for the production of an agent for inhibiting the binding of growth factors to tumor cells and especially for inhibiting a proli eration of tumor cells or tumors that are produced by growth factors .
  • An inhibitor of the progesterone receptor in terms of this invention is preferably a substance that competitively inhibits the binding of progesterone to its receptor.
  • the inhibitor of the progesterone receptor is preferably selected from 17 ⁇ -fluoroalkyl steroids, as
  • R 1 stands for a methyl or ethyl group
  • R 3 stands for a free, etherified, or esterified hydroxy group
  • R 4 and R 5 each stand for a hydrogen atom, together for an additional bond or a methylene group
  • St stands for a steroidal ABC-ring system of partial formula A, B or C
  • R b means a hydrogen atom, a straight - chain C j -C 4 alkyl group or a branched C 3 -C 4 alkyl group or a halogen atom,
  • R 7 means a hydrogen atom, a straight-chain C 1 -C 4 alkyl group or a branched C 3 -C 4 alkyl group, or, if St stands for a steroidal ABC-ring system A or B, in addition R 6 and R 7 together mean an additional bond,
  • R a means a radical Y or an aryl radical that is optionally substituted with a group Y in several places, whereby Y is a hydrogen atom, a halogen atom, an -OH, - N0 2 , -N-, -CN, -NR 9a R 9b , -NHS0 2 R 9 , -C0 2 R 9 , C x -C 10 alkoxy, C.-C 10 alkanoyloxy, benzoyloxy-C.-C 10 alkanoyl, C..-C 10 hydroxyalkyl or benzoyl group, and R 9a and R 9b are the same or different and like R 9 represent a hydrogen atom or a C ⁇ C- ⁇ alkyl group, and for radicals -NR 9a R 9b , also their physiologically compatible salts with acids and for radicals -C0 2 R 9 with R 9 in the meaning of hydrogen also their physiologically compatible salts with bases.
  • inhibitors of the progesterone receptor is the compound ll ⁇ - (4-acetylphenyl) -17 ⁇ -
  • hydroxypropyl) -13 ⁇ -estra-4 , 9-dien-3-one are also suitable, however .
  • the action of the progesterone receptor inhibitors is found especially in the case of tumor cells that have a high and/or constitutive progesterone receptor expression, for example the progesterone receptor-positive breast carcinoma cell line T47D (Sartorius et al . , Cancer Res. 54 (1994), 3668-3877).
  • the progesterone receptor inhibitors inhibit the progesterone-induced enhancement of the expression of growth factors, especially those factors that bind to growth factors of the EGF receptor family, such as, for example, the EGF receptor.
  • the inhibitors especially preferably inhibit the binding of EGF to human breast carcinoma cells.
  • the progesterone receptor inhibitors can therefore be used for tumor therapy in mammals and preferably in humans, specifically especially to block the progression of a tumor, especially a breast carcinoma of steroid- dependent growth to growth-factor-dependent growth.
  • an effective treatment of the tumor can take place in the stage of the steroid-dependent growth, e.g. by antiestrogens, without the tumor being able to progress in the stage of the growth- factor-dependent growth, associated with a considerable worsening of the prognosis for the patient.
  • the administration of the progesterone receptor inhibitors can also produce a slowing of tumor growth in the stage of the growth-factor-dependent growth.
  • non- steroidal antiestrogens such as, e.g., tamoxifen and nafoxidine, and raloxifene and EM800
  • the two last-mentioned antiestrogens are representatives of the thus mentioned SERMs (Selective Estrogen Receptor Modulators) ; also, other compounds with the profile of action of the SERMS can be used according to the invention, e.g., the compounds that are mentioned in PCT/EP99/05093 and of the latter in turn especially the compound 5- (4- ⁇ 5- [(RS) -4,4,5,5,5-pentafluoropentyl) sulfinyl] - pentyloxy ⁇ phenyl) -6-phenyl-8 , 9-dihydro-7H-benzocyclohepten-2-ol .
  • steroidal antiestrogens comprise those that are disclosed in EP 0 348 341 A, especially Faslodex, and those that are disclosed in WO98/07740, especially ll ⁇ -fluoro-7 ⁇ - ⁇ 5- [N- methyl-N-3- (4,4,5,5, 5-pentafluoropentylthio-propylamino] -pentyl ⁇ - estra-1, 3 , 5 (10) triene-3 , 17 ⁇ -diol, or those that are described in
  • W099/33855 especially ll ⁇ -fluoro-7 ⁇ - ⁇ 5- [methyl - (7, 7, 8, 8, 9, 9, 10, 10, 10-nonafluoro-decyl) -amino] -pentyl ⁇ -estra- 1, 3 , 5 (10) triene-3 , 17 ⁇ -diol or pharmaceutically compatible derivatives or analogs thereof.
  • Aromatase inhibitors with an antiestrogenic effect such as, for example, those that are known from pages 7-8 of EP 0 495 825 Bl, can likewise be used as antiestrogens .
  • the administration of the progesterone receptor inhibitors can be carried out according to commonly used methods, for example locally, topically, subcutaneously, enterally or parenterally .
  • enteral administration especially tablets, coated tablets, capsules, pills, suspensions or solutions are suitable, which can be produced in the usual way with the additives and vehicles that are known in gallenicals.
  • vaginal suppositories or transdermal systems such as skin patches are suitable.
  • the subcutaneous administration can be carried out by injection with an oily solution.
  • the daily dose of the active compound (s) is approximately 0.1 to 400 mg, preferably approximately 10-100 mg and especially approximately 50 mg.
  • Figure 1 shows the antiproliferative action of test substances on the breast carcinoma cell line T47D.
  • Figure 2 shows the amounts of protein of progesterone receptor (PR) and estrogen receptor (ER) in breast carcinoma cell line T47D.
  • Figure 3 shows the transcriptional activity of the progesterone receptor in T47D cells.
  • Figure 4 shows a Scatchard analysis of the binding of EGF to T47D cells as a function of the presence of test substances.
  • Figure 5 shows the dependence of the binding of EGF to T47D cells on the presence of test substances .
  • the human estrogen receptor (ER) - and progesterone receptor (PR) -positive breast carcinoma cell line T47D (Freake et al . , BBRC 101 (1981) , 1131-1138) was used.
  • the tumor cells were cultivated at 5000 cells/well in 96- well plates for 6 days in RPMI medium plus 10% bovine serum, 200 nM of insulin and 0.1 nM of estradiol in the presence of the compounds that are indicated in each case, and the growth was determined by staining with crystal violet.
  • the amounts of PR and ER in cell lysates are determined with use of steroid binding assays with radiolabeled progesterone or estradiol according to methods described in Fuhrmann et al . (Contraception 54 (1996) , 243-251) .
  • R5020-Pretreated T47D cells were incubated for 2 hours with X25 I-EGF at 4°C.
  • the unspecific binding was always less than 10% of the total binding.
  • T47D cells were transiently transfixed with MTV-LUC (Cato et al . , EMBO J., 9: 2237-40) and cultivated in the absence or the presence of 1 nM of R5020.
  • MTV-LUC Cato et al . , EMBO J., 9: 2237-40
  • the transiently transfixed T47D cells were treated with R5020 and in addition with increasing concentrations of compound A or RU486. After 24 hours, a luciferase test was performed.
  • Figure 1 shows the antiproliferative action of various test substances.
  • T47D cells were cultivated in the presence (upper cross-hatching) or absence (lower cross-hatching) of 0.1 nM of E 2 plus increasing concentrations of compound A (*), onapristone ( ⁇ ) , ZK191703 (•) or 4-OH-Tam ( ⁇ ) .
  • compound A also shows a significant antiproli erative action at extremely small concentrations.
  • Figure 2 shows the amounts of PR- and ER protein in T47D cells .
  • Figure 3 shows the transcriptional activity of PR in T47D cells, whereby the respective cells were transiently transfixed with MTV-LUC and cultivated (a) in the absence of (Co) or the presence of 1 nM of R5020.
  • the transiently transfixed T47D cells were treated with 0.1 nM of R5020 and increasing concentrations of compound A or RU468 (b) .
  • FIG 4 a Scatchard analysis of the 12S I-EGF binding to T47D cells is shown.
  • the cells were cultivated for 48 hours in the presence of 20 nM of R5020 with or without 20 nM of compound A and then washed. Then, the EGF-binding over a concentration range of 0.25 to 150 ng/ml of EGF was determined by incubation for 2 hours at 4°C.
  • the insertions show the amount of bonded ligands relative to the logarithm of the free ligand concentration. It is clear that it was possible to block the increase of the EGF binding that is caused by R5020 (middle figure) relative to monitoring (upper figure) when compound A (lower figure) is added.
  • a stimulation of the T47D cells with R5020 resulted in a 2x to 3x-increased EGF-receptor expression, which was blocked by compound A.
  • the binding of EGF to the cells was increased 2- to 3 -fold and could be prevented by compound A and less efficiently by onapristone.
  • the increased EGF-binding to R5020-treated cells could be produced by an enhanced EGF- receptor expression or increased heterodimer formation between the EGF receptor and erbB2.

Landscapes

  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Steroid Compounds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Compounds Of Unknown Constitution (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention relates to the use of progesterone receptor inhibitors for inhibition of growth-factor-dependency of tumor cells.

Description

Inhibition of the Growth Factor Dependency of Tumor Cells
Description
The invention relates to the use of progesterone receptor inhibitors for inhibiting the growth factor dependency of tumor cells .
Estradiol and progesterone are involved in the development of breast cancer. At the time of diagnosis, however, only about 1/3 of the tumors show a steroid hormone dependency. It is assumed that in the majority of steroid hormone-resistant tumors, the proliferation control for local -acting autocrine or paracrine peptidic growth factors is taken over. In this case, invasive tumors with extremely poor prognosis that are growth-factor- receptor-positive and steroid hormone-resistant result (Elledge et al . , Semin. Onkol . 19 (1992), 244-253).
Growth factors regulate the cell growth by activation of intracellular signal transduction pathways after binding to highly affine tyrosine kinase receptors on the cell surface. More recent findings suggest that breast carcinoma cells can be sensitized by progestins for the mitogenic action of EGF (Groshong et al . , Mol . Endocrinol . 11 (1997), 1593-1607). Thus, for example, it was possible for progesterone in the human breast carcinoma cell line T47D to induce the onset of cells in the S- phase accompanied by a transient increase of the activity of cyclin ID and the cyclin-dependent kinase 4. The growth stimulation is limited to a single cycle, however, and is followed by a growth arrest at the Gl/S-transition of the second cycle (Groshong et al . (1997), supra: Musgrove et al . , Mol . Cell. Biol. 13 (1993), 3577-3587). In its condition that is stopped by progesterone, the cells are sensitive to the proliferative action of EGF. In addition, it was shown that progesterone enhances the action of EGF on T47D cells by ramping up EGFR, Erb2 and Erb3 and increases the tyrosine phosphorylation of signal molecules (Lange et al., J. Biol. Chem. 273 (1998), 31308-31316; Richer et al . , J. Biol. Chem. 273 (1998), 31317-31326). In contrast, it has not yet been possible to show an inhibition of the action of EGF on tumor cells by influencing the progesterone receptor.
Within the scope of tests leading to this invention, it has now been found, surprisingly enough, that inhibitors of the progesterone receptor, e.g., 17α-fluoroalkyl steroids, can at least partially inhibit the binding of growth factors, such as EGF, to tumor cells, especially to tumor cells that have a high and/or constitutive expression of the progesterone receptor.
A subject of this invention is thus the use of an inhibitor of the progesterone receptor for the production of an agent for inhibiting the binding of growth factors to tumor cells and especially for inhibiting a proli eration of tumor cells or tumors that are produced by growth factors . An inhibitor of the progesterone receptor in terms of this invention is preferably a substance that competitively inhibits the binding of progesterone to its receptor. In this case, the inhibitor of the progesterone receptor is preferably selected from 17α-fluoroalkyl steroids, as
they are disclosed in, e.g., W098/34947. These 17α-fluoroalkyl steroids exhibit general formula I :
Figure imgf000004_0001
in which
R1 stands for a methyl or ethyl group,
R2 stands for a radical of formula CnFmH0, whereby n = 2,
3, 4, 5 or 6, m > 1 and m + o = 2n 4- 1, R3 stands for a free, etherified, or esterified hydroxy group, R4 and R5 each stand for a hydrogen atom, together for an additional bond or a methylene group, St stands for a steroidal ABC-ring system of partial formula A, B or C
Figure imgf000005_0001
B
ich
Rb means a hydrogen atom, a straight - chain Cj -C4 alkyl group or a branched C3 -C4 alkyl group or a halogen atom,
R7 means a hydrogen atom, a straight-chain C1-C4 alkyl group or a branched C3-C4 alkyl group, or, if St stands for a steroidal ABC-ring system A or B, in addition R6 and R7 together mean an additional bond,
X means an oxygen atom, a hydroxymino grouping = N — OH or two hydrogen atoms,
Ra means a radical Y or an aryl radical that is optionally substituted with a group Y in several places, whereby Y is a hydrogen atom, a halogen atom, an -OH, - N02, -N-, -CN, -NR9aR9b, -NHS02R9, -C02R9, Cx-C10 alkoxy, C.-C10 alkanoyloxy, benzoyloxy-C.-C10 alkanoyl, C..-C10 hydroxyalkyl or benzoyl group, and R9a and R9b are the same or different and like R9 represent a hydrogen atom or a C^C-^ alkyl group, and for radicals -NR9aR9b, also their physiologically compatible salts with acids and for radicals -C02R9 with R9 in the meaning of hydrogen also their physiologically compatible salts with bases.
An especially preferred example of such inhibitors of the progesterone receptor is the compound llβ- (4-acetylphenyl) -17β-
hydroxy-17α- (1,1,2,2, 2-pentafluoroethyl) -estra-4 , 9-dien-3 -one (compound A below) . Moreover, other antiprogestins, for example onapristone (llβ- [p- (dimethylamino) phenyl] -17α-hydroxy-17- (3-
hydroxypropyl) -13α-estra-4 , 9-dien-3-one) are also suitable, however .
The action of the progesterone receptor inhibitors is found especially in the case of tumor cells that have a high and/or constitutive progesterone receptor expression, for example the progesterone receptor-positive breast carcinoma cell line T47D (Sartorius et al . , Cancer Res. 54 (1994), 3668-3877). The progesterone receptor inhibitors inhibit the progesterone-induced enhancement of the expression of growth factors, especially those factors that bind to growth factors of the EGF receptor family, such as, for example, the EGF receptor. The inhibitors especially preferably inhibit the binding of EGF to human breast carcinoma cells. According to this invention, the progesterone receptor inhibitors can therefore be used for tumor therapy in mammals and preferably in humans, specifically especially to block the progression of a tumor, especially a breast carcinoma of steroid- dependent growth to growth-factor-dependent growth. In this way, an effective treatment of the tumor can take place in the stage of the steroid-dependent growth, e.g. by antiestrogens, without the tumor being able to progress in the stage of the growth- factor-dependent growth, associated with a considerable worsening of the prognosis for the patient. The administration of the progesterone receptor inhibitors can also produce a slowing of tumor growth in the stage of the growth-factor-dependent growth.
For the purpose of this invention, non- steroidal antiestrogens, such as, e.g., tamoxifen and nafoxidine, and raloxifene and EM800, can be used. The two last-mentioned antiestrogens are representatives of the thus mentioned SERMs (Selective Estrogen Receptor Modulators) ; also, other compounds with the profile of action of the SERMS can be used according to the invention, e.g., the compounds that are mentioned in PCT/EP99/05093 and of the latter in turn especially the compound 5- (4-{5- [(RS) -4,4,5,5,5-pentafluoropentyl) sulfinyl] - pentyloxy}phenyl) -6-phenyl-8 , 9-dihydro-7H-benzocyclohepten-2-ol .
Examples of steroidal antiestrogens comprise those that are disclosed in EP 0 348 341 A, especially Faslodex, and those that are disclosed in WO98/07740, especially llβ-fluoro-7α- {5- [N- methyl-N-3- (4,4,5,5, 5-pentafluoropentylthio-propylamino] -pentyl} - estra-1, 3 , 5 (10) triene-3 , 17β-diol, or those that are described in
W099/33855, especially llβ-fluoro-7α- { 5- [methyl - (7, 7, 8, 8, 9, 9, 10, 10, 10-nonafluoro-decyl) -amino] -pentyl} -estra- 1, 3 , 5 (10) triene-3 , 17β-diol or pharmaceutically compatible derivatives or analogs thereof. Aromatase inhibitors with an antiestrogenic effect, such as, for example, those that are known from pages 7-8 of EP 0 495 825 Bl, can likewise be used as antiestrogens .
The administration of the progesterone receptor inhibitors can be carried out according to commonly used methods, for example locally, topically, subcutaneously, enterally or parenterally . For enteral administration, especially tablets, coated tablets, capsules, pills, suspensions or solutions are suitable, which can be produced in the usual way with the additives and vehicles that are known in gallenicals. For local or topical use, for example, vaginal suppositories or transdermal systems such as skin patches are suitable. The subcutaneous administration can be carried out by injection with an oily solution.
A dosage unit can contain, for example, 0.1 to 100 mg of active compound (s) (= inhibitor (s) of the progesterone receptor).
For administration in humans, the daily dose of the active compound (s) is approximately 0.1 to 400 mg, preferably approximately 10-100 mg and especially approximately 50 mg.
In addition, the invention is to be explained by the following examples and figures. Here: Figure 1 shows the antiproliferative action of test substances on the breast carcinoma cell line T47D. Figure 2 shows the amounts of protein of progesterone receptor (PR) and estrogen receptor (ER) in breast carcinoma cell line T47D. Figure 3 shows the transcriptional activity of the progesterone receptor in T47D cells. Figure 4 shows a Scatchard analysis of the binding of EGF to T47D cells as a function of the presence of test substances. Figure 5 shows the dependence of the binding of EGF to T47D cells on the presence of test substances .
Example
1. Materials and Methods
Materials :
125I-EGF (100 mCi/mmol) was obtained by Amersham Buchler. Compound A, hydrotamoxifen (4-0H-Tam) , ZM182780 and estradiol were synthesized in the Institut fur Arzneimittelchemie [Institute for Pharmaceutical Agent Chemistry] of the Schering AG according to known methods.
Cell lines :
The human estrogen receptor (ER) - and progesterone receptor (PR) -positive breast carcinoma cell line T47D (Freake et al . , BBRC 101 (1981) , 1131-1138) was used.
Growth studies :
The tumor cells were cultivated at 5000 cells/well in 96- well plates for 6 days in RPMI medium plus 10% bovine serum, 200 nM of insulin and 0.1 nM of estradiol in the presence of the compounds that are indicated in each case, and the growth was determined by staining with crystal violet.
Amount of PR and ER protein:
The amounts of PR and ER in cell lysates are determined with use of steroid binding assays with radiolabeled progesterone or estradiol according to methods described in Fuhrmann et al . (Contraception 54 (1996) , 243-251) .
Binding of α25I-EGF to tumor cells:
R5020-Pretreated T47D cells were incubated for 2 hours with X25I-EGF at 4°C. The unspecific binding was always less than 10% of the total binding.
Transactivation assay:
T47D cells were transiently transfixed with MTV-LUC (Cato et al . , EMBO J., 9: 2237-40) and cultivated in the absence or the presence of 1 nM of R5020. In the test on a PR-mediated antagonism, the transiently transfixed T47D cells were treated with R5020 and in addition with increasing concentrations of compound A or RU486. After 24 hours, a luciferase test was performed.
2. Results
Figure 1 shows the antiproliferative action of various test substances. T47D cells were cultivated in the presence (upper cross-hatching) or absence (lower cross-hatching) of 0.1 nM of E2 plus increasing concentrations of compound A (*), onapristone (■) , ZK191703 (•) or 4-OH-Tam (♦) . In the case of T47D cells, compound A also shows a significant antiproli erative action at extremely small concentrations. Figure 2 shows the amounts of PR- and ER protein in T47D cells .
Figure 3 shows the transcriptional activity of PR in T47D cells, whereby the respective cells were transiently transfixed with MTV-LUC and cultivated (a) in the absence of (Co) or the presence of 1 nM of R5020. In the test for a PR-mediated antagonism, the transiently transfixed T47D cells were treated with 0.1 nM of R5020 and increasing concentrations of compound A or RU468 (b) .
In Figure 4, a Scatchard analysis of the 12SI-EGF binding to T47D cells is shown. The cells were cultivated for 48 hours in the presence of 20 nM of R5020 with or without 20 nM of compound A and then washed. Then, the EGF-binding over a concentration range of 0.25 to 150 ng/ml of EGF was determined by incubation for 2 hours at 4°C. The insertions show the amount of bonded ligands relative to the logarithm of the free ligand concentration. It is clear that it was possible to block the increase of the EGF binding that is caused by R5020 (middle figure) relative to monitoring (upper figure) when compound A (lower figure) is added.
In figure 5, the binding of 125I-EGF to intact T47D cells is shown. For this purpose, the cells were treated for 48 hours with 2 or 20 nM of R5020 plus compound A or- onapristone or compound A alone. It can also be seen here that compound A blocks the increase of the EGF-binding to T47D cells caused by R5020. A similar -- although considerably weaker effect -- is also found for onapristone. 3. Discussion
The above results show that the estradiol-stimulated growth of T47D cells with high and constitutive PR contact was effectively blocked by compound A.
By transactivation assays, it was possible to show that the PR was transcriptionally active in the T47D cells and could be blocked by compound 1.
A stimulation of the T47D cells with R5020 resulted in a 2x to 3x-increased EGF-receptor expression, which was blocked by compound A. At the same time, the binding of EGF to the cells was increased 2- to 3 -fold and could be prevented by compound A and less efficiently by onapristone. The increased EGF-binding to R5020-treated cells could be produced by an enhanced EGF- receptor expression or increased heterodimer formation between the EGF receptor and erbB2.
These results show the interactions between PR- and growth factor-signal systems in human breast carcinoma cells. By use of antiprogestins, the progression of tumor cells from steroid- dependent growth is inhibited or prevented for growth-factor- dependent growth.

Claims

Claims
1. Use of an inhibitor of the progesterone receptor for the production of an agent for the inhibition of the binding of growth factors to tumor cells and/or to a tumor.
2. Use according to claim 1, characterized in that a proliferation of the tumor cells and/or the tumor produced by growth factors is inhibited.
3. Use according to claim 1 or 2 , wherein the inhibitor of the progesterone receptor is selected from 17α-fluoroalkyl steroids of general formula I
Figure imgf000014_0001
I in which
R1 stands for a methyl or ethyl group,
R2 stands for a radical of formula CnFmH0, whereby n is 2,
3, 4, 5 or 6, m > 1 and m + o = 2n + 1, R3 stands for a free, etherified, or esterified hydroxy group, R4 and R5 each stand for a hydrogen atom, together for an additional bond or a methylene group, St stands for a steroidal ABC-ring system of partial formula A, B or C
Figure imgf000015_0001
A
ich
R means a hydrogen atom, a straight-chain C1-C4 alkyl group or a branched C3-C4 alkyl group or a halogen atom, R7 means a hydrogen atom, a straight-chain Cx-C4 alkyl group or a branched C3-C4 alkyl group, or, if St stands for a steroidal ABC-ring system A or B, in addition R6 and R7 together mean an additional bond,
X means an oxygen atom, a hydroxymino grouping = N — OH or two hydrogen atoms,
R8 means a radical Y or an aryl radical that is optionally substituted with a group Y in several places, whereby Y is a hydrogen atom, a halogen atom, an -OH, -N02, -N3,
-CN, -NR9aR9b, -NHS02R9, -C02R9, C--C10 alkoxy, C1-C1 alkanoyloxy, benzoyloxy-Cα-C10 alkanoyl, C1-C10 hydroxyalkyl or benzoyl group, and Ra and R9b are the same or different and like R9 represent a hydrogen atom or a C^C^ alkyl group, and for radicals -NR9aR9b, also their physiologically compatible salts with acids and for radicals -C02R9 with R9 in the meaning of hydrogen also their physiologically compatible salts with bases.
4. Use according to claim 3, wherein the inhibitor of the progesterone receptor is the compound llβ- (4-acetylphenyl) -17β-
hydroxy-17α- (1,1,2,2, 2-pentafluoroethyl) estra-4 , 9-dien-3-one .
5. Use according to one of claims 1 to 4 , wherein the tumor cells have a high and/or constitutive progesterone receptor expression.
6. Use according to one of claims 1 to 5, wherein the tumor cells are breast carcinoma cells.
7. Use according to one of claims 1 to 6, wherein the binding of EGF and/or other factors, which bind to the EGF receptor, is inhibited in tumor cells.
8. Use according to one of claims 1 to 7, wherein the formation of heterodimers between the EGF receptor and erbB2 is inhibited .
9. Use according to one of claims 1 to 8 for tumor therapy.
10. Use according to claim 9 to inhibit the progression of a tumor of steroid-dependent growth to growth-factor-dependent growth .
PCT/EP2001/012004 2000-10-18 2001-10-17 Use of inhibitors of progesterone receptor for treating cancer Ceased WO2002032429A2 (en)

Priority Applications (25)

Application Number Priority Date Filing Date Title
DE60123138T DE60123138T2 (en) 2000-10-18 2001-10-17 USE OF PROGESTERONE RECEPTOR TREATMENT FOR THE TREATMENT OF STEROID-RESISTANT BREAST CANCER
PL01366415A PL366415A1 (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
HR20030386A HRP20030386A2 (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
JP2002535667A JP4335525B2 (en) 2000-10-18 2001-10-17 Growth factor-dependent inhibition of tumor cells
EA200300468A EA005945B1 (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
SK475-2003A SK4752003A3 (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
KR10-2003-7005358A KR20030044006A (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
MXPA03002953A MXPA03002953A (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells.
HU0301461A HUP0301461A3 (en) 2000-10-18 2001-10-17 Use of 17 alpha-fluoroalkyl steroids for producing pharmaceutical compositions for inhibition of the growth factor dependency of tumor cells
DK01987667T DK1414465T3 (en) 2000-10-18 2001-10-17 Inhibition of tumor cell growth factor dependence
EEP200300156A EE200300156A (en) 2000-10-18 2001-10-17 Inhibition of tumor cell growth factor dependence
EP01987667A EP1414465B1 (en) 2000-10-18 2001-10-17 Use of inhibitors of progesterone receptor for treating steroid resistant breast cancer
AU2002215958A AU2002215958B2 (en) 2000-10-18 2001-10-17 Use of inhibitors of progesterone receptor for treating cancer
US10/399,318 US20040157811A1 (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
AU1595802A AU1595802A (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
IL15497401A IL154974A0 (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
SI200130665T SI1414465T1 (en) 2000-10-18 2001-10-17 Use of inhibitors of progesterone receptor for treating steroid resistant breast cancer
UA2003054403A UA76128C2 (en) 2000-10-18 2001-10-17 Use of progesterone receptor inhibitors for treating steroid-resistant breast cancer
CA002423013A CA2423013A1 (en) 2000-10-18 2001-10-17 Use of inhibitors of progesteron receptor for treating cancer
BR0114677-7A BR0114677A (en) 2000-10-18 2001-10-17 Inhibition of tumor cell growth factor dependence
HK05104537.7A HK1071697B (en) 2000-10-18 2001-10-17 Inhibition of the growth factor dependency of tumor cells
IL154974A IL154974A (en) 2000-10-18 2003-03-18 Inhibition of the growth factor dependency of tumor cells
NO20031745A NO20031745L (en) 2000-10-18 2003-04-15 Inhibition of growth factor dependence on tumor cells
BG107745A BG107745A (en) 2000-10-18 2003-04-18 Inhibition of the growth factor dependency of tumor cells
CY20061101786T CY1106289T1 (en) 2000-10-18 2006-12-12 USE OF PROGESTEPONE RECEPTOR INHIBITORS IN THE TREATMENT OF STEPID-RESISTANT BREAST CANCER

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US24101000P 2000-10-18 2000-10-18
DE10051609A DE10051609A1 (en) 2000-10-18 2000-10-18 Use of an inhibitor of the progesterone receptor for the production of an agent for the inhibition of the binding of growth factors to tumor cells or to a tumor
US60/241,010 2000-10-18
DE10051609.2 2000-10-18

Publications (2)

Publication Number Publication Date
WO2002032429A2 true WO2002032429A2 (en) 2002-04-25
WO2002032429A3 WO2002032429A3 (en) 2004-02-19

Family

ID=26007397

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2001/012004 Ceased WO2002032429A2 (en) 2000-10-18 2001-10-17 Use of inhibitors of progesterone receptor for treating cancer

Country Status (25)

Country Link
EP (1) EP1414465B1 (en)
JP (1) JP4335525B2 (en)
KR (1) KR20030044006A (en)
CN (1) CN1311833C (en)
AT (1) ATE339208T1 (en)
AU (2) AU1595802A (en)
BG (1) BG107745A (en)
BR (1) BR0114677A (en)
CA (1) CA2423013A1 (en)
CY (1) CY1106289T1 (en)
CZ (1) CZ299945B6 (en)
DE (1) DE60123138T2 (en)
DK (1) DK1414465T3 (en)
EA (1) EA005945B1 (en)
EE (1) EE200300156A (en)
ES (1) ES2272561T3 (en)
HR (1) HRP20030386A2 (en)
HU (1) HUP0301461A3 (en)
IL (2) IL154974A0 (en)
MX (1) MXPA03002953A (en)
NO (1) NO20031745L (en)
NZ (1) NZ545965A (en)
PL (1) PL366415A1 (en)
SK (1) SK4752003A3 (en)
WO (1) WO2002032429A2 (en)

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8053426B2 (en) * 2006-11-15 2011-11-08 Bayer Schering Pharma Ag Progesterone receptor antagonists
US8278469B2 (en) 2009-07-20 2012-10-02 Bayer Pharma Aktiengesellschaft 17-hydroxy-17-pentafluoroethyl-estra-4,9(10)-dien-11-aryl derivatives, method of production thereof and use thereof for the treatment of diseases
US9085603B2 (en) 2010-02-10 2015-07-21 Bayer Intellectual Property Gmbh Progesterone receptor antagonists
US9096639B2 (en) 2009-07-20 2015-08-04 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluorethyl-estra-4,9(10)-dien-11-acyloxyalkylene phenyl derivatives, methods for the production thereof and use thereof for treating diseases
US9096640B2 (en) 2009-07-20 2015-08-04 Bayer Intellectual Property 17-hydroxy-17-pentafluoroethyl-estra-4,9(10)-diene-11-methylene oxyalkylene aryl derivatives, process for preparation thereof, and use thereof for treatment of diseases
US9102701B2 (en) 2009-07-21 2015-08-11 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluoroethyl-estra-4,9(10)-diene-11-ethynylphenyl derivatives, methods for the production thereof and use thereof for treating diseases
US9109004B2 (en) 2010-02-10 2015-08-18 Bayer Intellectual Property Gmbh Progesterone receptor antagonists
US9156877B2 (en) 2009-07-20 2015-10-13 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluorethyl-estra-4,9(10)-dien-11-benzylidene derivatives, methods for the production thereof and use thereof for treating diseases
US9206219B2 (en) 2009-07-21 2015-12-08 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluorethyl-estra-4,9(10)-dien-11-aryl derivatives, methods for the production thereof and the use thereof for treating diseases

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0466315A3 (en) * 1990-05-30 1992-07-01 Larrian Gillespie Compositions containing xylan sulphates for affecting growth factor function and for inhibiting fibroblast proliferation
US5238950A (en) * 1991-12-17 1993-08-24 Schering Corporation Inhibitors of platelet-derived growth factor
DE19706061A1 (en) * 1997-02-07 1998-08-13 Schering Ag Anti-gestagen effective steroids with fluorinated 17alpha alkyl chain
AR015500A1 (en) * 1997-12-23 2001-05-02 Schering Ag 11 BETA-HALOGEN-STRATRIENS REPLACED IN 7 ALPHA, PROCEDURE FOR PREPARING PHARMACEUTICAL PREPARATIONS THAT CONTAIN SUCH 11 BETA-HALOGEN-STRATRIENS REPLACED IN 7 ALPHA, AS WELL AS USED IN THE PREPARATION OF MEDICINES.
FR2789393B1 (en) * 1999-02-05 2001-10-12 Centre Nat Rech Scient STEROID LACTOL

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8053426B2 (en) * 2006-11-15 2011-11-08 Bayer Schering Pharma Ag Progesterone receptor antagonists
US8278469B2 (en) 2009-07-20 2012-10-02 Bayer Pharma Aktiengesellschaft 17-hydroxy-17-pentafluoroethyl-estra-4,9(10)-dien-11-aryl derivatives, method of production thereof and use thereof for the treatment of diseases
US9096639B2 (en) 2009-07-20 2015-08-04 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluorethyl-estra-4,9(10)-dien-11-acyloxyalkylene phenyl derivatives, methods for the production thereof and use thereof for treating diseases
US9096640B2 (en) 2009-07-20 2015-08-04 Bayer Intellectual Property 17-hydroxy-17-pentafluoroethyl-estra-4,9(10)-diene-11-methylene oxyalkylene aryl derivatives, process for preparation thereof, and use thereof for treatment of diseases
US9156877B2 (en) 2009-07-20 2015-10-13 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluorethyl-estra-4,9(10)-dien-11-benzylidene derivatives, methods for the production thereof and use thereof for treating diseases
US10155004B2 (en) 2009-07-20 2018-12-18 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluoroethyl-estra-4,9(10)-dien-11-aryl derivatives, method of production thereof and use thereof for the treatment of diseases
US9102701B2 (en) 2009-07-21 2015-08-11 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluoroethyl-estra-4,9(10)-diene-11-ethynylphenyl derivatives, methods for the production thereof and use thereof for treating diseases
US9206219B2 (en) 2009-07-21 2015-12-08 Bayer Intellectual Property Gmbh 17-hydroxy-17-pentafluorethyl-estra-4,9(10)-dien-11-aryl derivatives, methods for the production thereof and the use thereof for treating diseases
US9085603B2 (en) 2010-02-10 2015-07-21 Bayer Intellectual Property Gmbh Progesterone receptor antagonists
US9109004B2 (en) 2010-02-10 2015-08-18 Bayer Intellectual Property Gmbh Progesterone receptor antagonists

Also Published As

Publication number Publication date
AU2002215958C1 (en) 2002-04-29
AU1595802A (en) 2002-04-29
HUP0301461A3 (en) 2010-01-28
IL154974A0 (en) 2003-10-31
WO2002032429A3 (en) 2004-02-19
SK4752003A3 (en) 2003-10-07
ATE339208T1 (en) 2006-10-15
EE200300156A (en) 2003-08-15
MXPA03002953A (en) 2003-08-07
AU2002215958B2 (en) 2006-08-17
JP4335525B2 (en) 2009-09-30
PL366415A1 (en) 2005-01-24
HK1071697A1 (en) 2005-07-29
CZ299945B6 (en) 2008-12-29
NO20031745D0 (en) 2003-04-15
IL154974A (en) 2007-05-15
DE60123138D1 (en) 2006-10-26
DE60123138T2 (en) 2007-09-13
CN1311833C (en) 2007-04-25
EA200300468A1 (en) 2003-10-30
JP2004513094A (en) 2004-04-30
BG107745A (en) 2004-01-30
ES2272561T3 (en) 2007-05-01
EP1414465B1 (en) 2006-09-13
CA2423013A1 (en) 2002-04-25
CY1106289T1 (en) 2011-10-12
HRP20030386A2 (en) 2005-04-30
HUP0301461A2 (en) 2003-10-28
CN1551772A (en) 2004-12-01
NO20031745L (en) 2003-04-15
NZ545965A (en) 2007-09-28
EP1414465A2 (en) 2004-05-06
KR20030044006A (en) 2003-06-02
BR0114677A (en) 2003-10-07
DK1414465T3 (en) 2007-01-29
EA005945B1 (en) 2005-08-25

Similar Documents

Publication Publication Date Title
US8735381B2 (en) Progesterone antagonists such as CDB-4124 in the treatment of endometriosis, uterine fibroids, dysmenorrhea, breast cancer, etc
Sutherland et al. Effect of medroxyprogesterone acetate on proliferation and cell cycle kinetics of human mammary carcinoma cells
Sherwin et al. Sex steroids and affect in the surgical menopause: a double-blind, cross-over study
Kopp et al. Transforming growth factor β2 (TGF-β2) levels in plasma of patients with metastatic breast cancer treated with tamoxifen
AU2007327707A1 (en) Compositions and methods for suppressing endometrial proliferation
US20040157811A1 (en) Inhibition of the growth factor dependency of tumor cells
EP1414465B1 (en) Use of inhibitors of progesterone receptor for treating steroid resistant breast cancer
AU2002215958A1 (en) Use of inhibitors of progesterone receptor for treating cancer
Quinn Hormonal treatment of endometrial cancer
Lentz Endocrine therapy of endometrial cancer
HK1071697B (en) Inhibition of the growth factor dependency of tumor cells
UA76128C2 (en) Use of progesterone receptor inhibitors for treating steroid-resistant breast cancer
Clarke et al. 1986) and calmodulin (Lam, 1984) is inhibited by Tamoxifen as a result of direct interactions. Structural parameters of the plasma
Clarke et al. influence its biological activity and that, when significantly altered, could contribute to the emergence of an antiestrogen resistant phenotype. These
Banerjee The combination of VEGF inhibitors and anti-oestrogen therapies in breast cancer

Legal Events

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

Ref document number: P-300/03

Country of ref document: YU

AK Designated states

Kind code of ref document: A2

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PH PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 154974

Country of ref document: IL

WWE Wipo information: entry into national phase

Ref document number: 2423013

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: PA/a/2003/002953

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 525173

Country of ref document: NZ

WWE Wipo information: entry into national phase

Ref document number: 381/MUMNP/2003

Country of ref document: IN

WWE Wipo information: entry into national phase

Ref document number: 2002535667

Country of ref document: JP

WWE Wipo information: entry into national phase

Ref document number: 1-2003-500261

Country of ref document: PH

WWE Wipo information: entry into national phase

Ref document number: 4752003

Country of ref document: SK

Ref document number: PV2003-1071

Country of ref document: CZ

Ref document number: 018173853

Country of ref document: CN

WWE Wipo information: entry into national phase

Ref document number: 1020037005358

Country of ref document: KR

ENP Entry into the national phase

Ref document number: 10774501

Country of ref document: BG

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 2001987667

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 200300468

Country of ref document: EA

WWE Wipo information: entry into national phase

Ref document number: P20030386A

Country of ref document: HR

WWE Wipo information: entry into national phase

Ref document number: 2003/03785

Country of ref document: ZA

Ref document number: 200303785

Country of ref document: ZA

Ref document number: 2002215958

Country of ref document: AU

Ref document number: 03040572

Country of ref document: CO

WWP Wipo information: published in national office

Ref document number: 1020037005358

Country of ref document: KR

WWP Wipo information: published in national office

Ref document number: PV2003-1071

Country of ref document: CZ

WWE Wipo information: entry into national phase

Ref document number: 10399318

Country of ref document: US

WWP Wipo information: published in national office

Ref document number: 2001987667

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 2001987667

Country of ref document: EP