WO1997043268A1 - Derives de benzodizocine a substitution 8-hydrocarbyle, leur preparation et leur utilisation comme modulateurs de la proteine kinase c (pkc) - Google Patents

Derives de benzodizocine a substitution 8-hydrocarbyle, leur preparation et leur utilisation comme modulateurs de la proteine kinase c (pkc) Download PDF

Info

Publication number
WO1997043268A1
WO1997043268A1 PCT/US1997/008141 US9708141W WO9743268A1 WO 1997043268 A1 WO1997043268 A1 WO 1997043268A1 US 9708141 W US9708141 W US 9708141W WO 9743268 A1 WO9743268 A1 WO 9743268A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
compound
mixture
pharmaceutically acceptable
formula
Prior art date
Application number
PCT/US1997/008141
Other languages
English (en)
Inventor
Alan P. Kozikowski
Dawei Ma
Robert I. Glazer
Original Assignee
Georgetown University
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 Georgetown University filed Critical Georgetown University
Priority to EP97925564A priority Critical patent/EP0918757A1/fr
Priority to AU30666/97A priority patent/AU3066697A/en
Priority to JP09541093A priority patent/JP2000510149A/ja
Publication of WO1997043268A1 publication Critical patent/WO1997043268A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D245/00—Heterocyclic compounds containing rings of more than seven members having two nitrogen atoms as the only ring hetero atoms
    • C07D245/04—Heterocyclic compounds containing rings of more than seven members having two nitrogen atoms as the only ring hetero atoms condensed with carbocyclic rings or ring systems
    • C07D245/06—Heterocyclic compounds containing rings of more than seven members having two nitrogen atoms as the only ring hetero atoms condensed with carbocyclic rings or ring systems condensed with one six-membered ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • PKC protein kinase C
  • the PKC gene family consists presently of 1 1 genes which are divided into four subgroups: 1) classical PKC ⁇ , ⁇ ,, ⁇ 2 ( ⁇ , and ⁇ 2 are alternately spliced forms ofthe same gene) and ⁇ , 2) novel PKC ⁇ , e, ⁇ , and ⁇ , 3) atypical PKC ⁇ , ⁇ , ⁇ and i and 4) PKC ⁇ .
  • PKC ⁇ resembles the novel PKC isoforms but 5 differs by having a putative transmembrane domain (reviewed in Blobe et al., Cancer Metast. Rev.. 13, 41 1 (1994)); Hug et al., Biochem J.. 291. 329 (1993); Kikkawa et al., Ann. Rev. Biochem.
  • the ⁇ , ⁇ character ⁇ 2 and ⁇ isoforms are Ca 2+ , phospholipid- and diacylglycerol-dependent and represent the classical isoforms of PKC, whereas the other isoforms are activated by 0 phospholipid and diacylglycerol but are not dependent on Ca 2 '. All isoforms encompass 5 variable (VI -V5) regions and the a, ⁇ and ⁇ isoforms contain four (C1-C4) structural domains which are highly conserved.
  • All isoforms except PKC ⁇ , ⁇ , and ⁇ lack the C2 domain, and the ⁇ , ⁇ and i isoforms also lack one of two cysteine-rich zinc finger domains in Cl to which diacylglycerol binds.
  • the Cl domain also contains the pseudo substrate sequence which is highly conserved among all isoforms, and which serves an autoregulatory function by blocking the substrate-binding site to produce an inactive conformation ofthe enzyme (House et al. Science. 238. 1726 ( 1987)).
  • PKC isoforms are thought to have highly specialized roles in signal transduction in response to physiological stimuli (Nishizuka, Cancer. 10. 1892 (1989)), as well as in neoplastic transformation and differentiation (Glazer, Protein Kinase C. J. F. Kuo, ed., Oxford U. Press (1994) at pages 171-198).
  • PKC has served as a focal point for the design of anticancer drugs (Gescher, Brit. J. Cancer. 66. 10 (1992)).
  • Antisense expression of either the PKC ⁇ cDNA (Ahmad et al., Neurosurgery. 35- 904 (1994)) or a phosphorothioate oligodeoxynucleotide (S- oligo) for PKC ⁇ has shown the efficacy of targeting PKC to inhibit the proliferation of A549 lung carcinoma cells (Dean et al., J. Biol. Chem.. 269. 16416 (1994)) and U-87 glioblastoma cells.
  • Selective cancer cell killing may be achieved either through the targeting of those isoforms found to be overexpressed in the cancer cells, or through the synergistic interaction of a cytotoxic drug like 1- ⁇ -D-arabinofuranosylcytosine with an appropriate PKC- based signaling interceptor.
  • Teleocidin was first isolated from the mycelia of Streptomyces mediocidicus as a mixture of highly toxic compounds by Takahashi et al., Bull. Agr. Chem. Soc. Japan. 24. 647 (1960). The structure of one of these metabolites was assigned by Hirata as shown by Figure 2, formula 1.
  • the lyngbyatoxin series can be obtained together with the teleocidin B group from Streptomyces mediocidicus as disclosed by S. Sakai et al., Tetrahedron Lett.. 27. 5219 (1986). Therefore, as depicted in Figure 2, they were named as teleocidin A-l (2a) and A-2 (2b) by Sakai. Indolactam V (3, ILV), which contains the basic ring structure ofthe teleocidins, is the simplest member ofthe family, and is produced in large quantities by actinomycetes strain NA34- 17 ( Figure 2).
  • TPA 12-O-tetradecanoylphorbol-13-acetate
  • R is H, (C,-C 5 )alkyl, OR", SR a , N(R a )(R b ), halo, NO 2 , NHC(O)[(C,-C 4 )alkyl] or NHOH;
  • R 2 is a (C 5 -C 22 ) hydrocarbyl group, optionally comprising 1 -3 double bonds, 1-2 triple bonds or a mixture thereof, or (C 6 -C 12 )aryl(C 2 -C 10 )alkyl, wherein the alkyl moiety optionally comprises 1-2 double bonds, 1-2 triple bonds or a mixture thereof; wherein said (C 5 -C 22 ) hydrocarbyl group or said (C 6 - C 12 )aryl(C 2 -C 10 )alkyl may optionally be substituted with 1 or 2 substituents independently selected from the group consisting of halo, hydroxy, cyano, nitro, (C,-C 5 )alkyl, (C,-C 5 )alkoxy, trifluoromethyl, trifluoromethoxy, -C(-O)O(C,-
  • R 3 is H, OH or halo;
  • R a and R b are independently H or (C,-C 5 )alkyl;
  • R c is a (C 5 -C 22 ) hydrocarbyl group;
  • R d is H or (C,-C 5 )alkyl;
  • R c and R f are independently hydrogen, (C,-C 5 )alkyl, or (C,-C 5 )alkanoyl, or together with the nitrogen to which they are attached are pyrrolidino, piperidino or morpholino; Z is H or (C,-C 5 )alkyl; and
  • Y is H or (C r C 5 )alkyl; or a pharmaceutically acceptable salt thereof.
  • compositions comprising an amount of one or more compounds of formula (I) effective to treat mammalian conditions associated with pathological cellular proliferation, particularly human cancers, such as solid tumors and leukemias, are also an embodiment ofthe invention.
  • the present invention also provides a method to inhibit the pathological proliferation of mammalian cells, such as cancer cells, by administering to a mammal afflicted with such a condition, an effective inhibitory amount of one or more ofthe compounds of formula I, preferably formulated as said pharmaceutical composition, i.e., in unit dosage form.
  • Novel intermediates and processes to prepare compounds of formula (I), as depicted in Figures 6-12 are also embodiments ofthe invention.
  • FIG. 1 shows the structural organization of the PKC gene family.
  • FIG. 2 shows the structures of telocidin B-4, A-l , A-2, and indolactam V.
  • FIG. 3 shows compounds ofthe invention.
  • FIG. 4 shows an electrophile useful for preparing compounds ofthe invention.
  • FIG. 5 shows an electrophile useful for preparing compounds ofthe invention.
  • FIG. 6 shows a scheme for preparing compounds of the invention.
  • FIG. 7 shows a scheme for preparing compounds ofthe invention.
  • FIG. 8 shows a scheme for preparing compounds ofthe invention.
  • FIG. 9 shows a scheme for preparing compounds ofthe invention.
  • FIG. 10 shows a scheme for preparing compounds ofthe invention.
  • FIG. 11 shows a scheme for preparing compounds ofthe invention.
  • FIG. 12 shows a scheme for preparing compounds ofthe invention.
  • FIG. 13 shows the cytotoxicity of compound 17 and ILV in MCF-7 and
  • FIG. 14 shows the Western blot of PKC isoform levels 24 hours after treatment with compound 17.
  • FIG. 15 shows the antitumor activity of compound 17 against the MDA-MB-231 xenograft in nude mice.
  • Halo is fluoro, chloro, bromo, or iodo.
  • alkyl encompasses branched or unbranched alkyl, cycloalkyl or (cycloalkyl)alkyl, but reference to an individual radical such as "propyl” embraces only the straight chain radical, a branched chain isomer such as "isopropyl” being specifically referred to.
  • Aryl comprises a phenyl radical, an ortho-fused bicyclic carbocyclic radical having about nine to ten ring atoms in which at least one ring is aromatic, as well as simple (C,-C 4 ) n alkylaryl wherein n is 1-3.
  • (C,-C 5 )alkyl is methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, cyclopropyl, cyclopropylmethyl, cyclobutyl, or cyclopentyl; and aryl is phenyl, methylphenyl, ethylphenyl, propylphenyl, dimethylphenyl, diethylphenyl, indenyl, methylindenyl, dimethylindenyl, naphthyl, methylnaphthyl, or dimethylnaphthyl.
  • a specific value for R is OR a , SR a , N(R a )(R b ), halo, NO 2 , NHC(O)[(C,-C 4 )alkyl] or NHOH; for R 2 is 1-decynyl or decyl; for R 3 is H; for R c is (C 5 -C 15 )alkyl; for Y is H; and for Z is methyl.
  • a more specific value for R is OR a .
  • Another specific group of compounds are compounds of formula I wherein Z is CH 3 ; Y is H; R, is OR a , SR a , N(R a )(R b ), halo, NO 2 , NHC(O)[(C,- C 4 )alkyl] or NHOH; R 2 is (C 5 -C, 5 )alkyl, optionally comprising 1-3 double bonds, 1-2 triple bonds or a mixture thereof; and R 3 is H; or a pharmaceutically acceptable salt thereof.
  • R is OR ⁇ SR a , N(R a )(R b ), halo, NO 2 , NHC(O)[(C,-C 4 )aIkyl] or NHOH; and R 2 is a hydrophobic (C 5 -C 22 ) hydrocarbyl group, optionally comprising 1-3 double bonds, 1-2 triple bonds or a mixture thereof, or (C 6 - C
  • R 2 is (C 5 -C 15 )alkyl, optionally comprising 1-3 double bonds, 1-2 triple bonds or a mixture thereof; or a pharmaceutically acceptable salt thereof.
  • R 2 is (C 6 -C ]2 )aryl(C 2 -C 10 )alkyl, wherein the alkyl moiety optionally comprises 1 -2 double bonds, 1 -2 triple bonds or a mixture thereof; or a pharmaceutically acceptable salt thereof.
  • a preferred group of compounds are compounds of formula I wherein Z is CH 3 ; Y is H; R 2 is (C 5 -C, 5 )alkyl, optionally comprising 1-3 double bonds, 1-2 triple bonds or a mixture thereof; and R 3 is H; or a pharmaceutically acceptable salt thereof.
  • Another preferred group of compounds are compounds of formula I wherein Z is CH 3 ; Y is H; R 2 is (C 5 -C, 5 )alkyl, optionally comprising 1-3 double bonds, 1-2 triple bonds or a mixture thereof; and R 3 is H; or a pharmaceutically acceptable salt thereof.
  • Another preferred group of compounds are compounds of formula I wherein Z is CH 3 ; Y is H; R 2 is (C 5 -C, 5 )alkyl, optionally comprising 1-3 double bonds, 1-2 triple bonds or a mixture thereof; and R 3 is H; or a pharmaceutically acceptable salt thereof.
  • Another preferred group of compounds are compounds of formula I wherein Z is CH 3 ; Y is H; R 2 is
  • R 2 is 1-decynyl; or a pharmaceutically acceptable salt thereof.
  • Compounds of formula I wherein R, is 1 -alkynyl can be prepared from a corresponding compound wherein R, is iodide by coupling with the requisite alkyne using a suitable catalyst, such as for example palladium. Suitable conditions for such a coupling reaction are illustrated in Example 1.
  • Compounds of formula I wherein R, is 1 -alkyl can be prepared from a corresponding compound wherein R, is 1 -alkynyl by hydrogenation of the alkyne bond using a suitable catalyst, such as for example palladium on carbon. Suitable conditions for such a hydrogenation are illustrated in Example 2.
  • Compounds of formula I can generally be prepared by the reaction of 2,6-disubstituted arylmetal compounds as nucleophiles with enantiomerically pure three carbon electrophiles inco ⁇ orating the necessary amino and hydroxyl groups in protected form.
  • Electrophiles Two readily available electrophiles ( Figures 4 and 5) are suitable for preparing compounds ofthe invention, the protected aziridinemethanol 21, and the protected serine aldehyde 23 ("Enantiospecific Synthesis of D- ⁇ , ⁇ -Diaminoalkanoic Acids" Beaulieu, P. L.; Schiller, P. W. Tetrahedron Lett. 1988, 29, 2019-2022).
  • the known L-serine-derived methyl or benzyl esters 20 can be reduced to the aziridinemethanol, e.
  • N-alkylarylamines have been successfully ortho-metalated by N-lithiation, reacted with CO 2 to form the lithium carbamate, and further treated with tert-butyllithium, this procedure failed in the case of aniline ("Carbon Dioxide: A Reagent for the Simultaneous Protection of Nucleophilic Centers and the Activation of Alternative Locations to Electrophilic Attack. 17. Substitution of N-Methyl-1- and
  • N-Methyl-2-naphthylamine and Side-Chain Functionalization of o-Toluidine Katritzky, A. R.; et al. J. Org. Chem. 1991, 56, 5045-5048.
  • N-(methoxycarbonyl)-O-(methoxymethyl)-w-aminophenol has been reported to undergo directed metalation mainly in the 2-position, whereas the corresponding ⁇ -Boc derivative reacted on the opposite side ofthe nitrogen in position 4 ( "Biosynthesis of Sarubicin A. Synthesis and Inco ⁇ oration of 6-Hydroxy[ 13 CO ,5 ⁇ H 2 ]anthranilamide" Gould, S. J.; Eisenberg, R. L.
  • N-(allyloxycarbonyl) derivative 24 ( Figure 6) is a convenient starting material for preparing compounds ofthe invention. While the allyloxycarbonyl group is not bulky, it is readily removed by various nucleophiles or hydride donors in the presence of a Pd catalyst.
  • An intermediate of formula 28 is particularly useful for preparing compounds of formula I.
  • An intermediate of formula 28 can be prepared as shown in Figure 6 by reaction of a nucleophile of formula 24 and an aziridine (electrophile) of formula 21 followed by deprotection ofthe aniline nitrogen.
  • the resulting aniline 25 can be alkylated to give a compound of formula 26. Hydrogenation of 26 followed by lactam formation yields an intermediate of formula 28.
  • the methoxymethoxy substituent in intermediate 28 provides access not only to 7-hydroxy- and 7-alkoxybenzolactams but, as discussed below, to a variety of other compounds ofthe invention via aryl triflate chemistry.
  • Benzolactams containing 7-halo or 7-CF 3 substituents can conveniently be synthesized from the requsite N-(allyloxycarbonyl)-w- substituted anilines using a procedure similar to the one described above.
  • a variety of potentially suitable substrates have been reported which differ in their thermal stability as well as in the ease or difficulty with which the second substituent can be transformed into NH 2 .
  • 3-Chlorobenzonitrile and N-tert-butyl-3-chlorobenzamide undergo directed lithiation in position 2 at -70°C, and the resulting organolithiums can be trapped with an electrophile ("Heteroatom-Facilitated Lithiations" Gschwend, H. W.; Rodriguez, H. R. Org. React. 1979, 26, 1-360 (unpublished results by Rodriguez, H. R.)).
  • 3-Fluoro- and 3-chIorophenyloxazolines have been metalated in position 2 ("A New Route to 3-Hydroxyphthalides: Application to the Synthesis of Racemic f5- ,3 C]
  • Lithiated 3-fluoro- and 3-chlorobenzaldehyde dimethyl acetal 35a, b can be reacted with aldehyde 23. Separation ofthe resulting stereoisomers followed by acid hydrolysis of the aryllithium addition product 36, yields an aldehyde, which forms a hemiacetal with the benzylic hydroxyl group (37).
  • Benzylic alcohol 37 can be selectively oxidized to a lactone using, for example, MnO 2 . Acetonide protection can then be restored. The lactam can be ammonolyzed to yield 39, and the liberated benzylic hydroxyl group protected by silylation.
  • a compound of formula I wherein R 3 is hydrogen can be prepared from a corresponding compound of formula I wherein R 3 is hydroxy, such as for example a compound of formula 42 , by formation of a cyclic thionocarbonate of formula 44 followed by Barton deoxygenation ("Synthesis of Deoxysugars and Deoxynucleosides from Diol Thiocarbonates" Barton, D. H. R.; Subramanian, R. J. Chem. Soc., Chem. Commun. 1976, 867-868).
  • the intermediate aryl triflate 46 can be used to prepare compounds of formula I having a variety of 7-substituents.
  • Methoxycarbonylation of 46 ( "Palladium Catalysed Alkoxycarbonylation of Phenols to Benzoate Esters" Dolle, R. E.; et al. J. Chem. Soc., Chem. Commun. 1987, 904-905) gives the ester 47 which can be transformed into the derivatives 48 and subsequently compounds of formula 49, by selective reduction of the ester moiety, followed by cuprate alkylation of the derived triflate.
  • the ester function of a compound of formula 47 can also be used to introduce a nitrogen atom into position 7 by means of a Curtius degradation.
  • the product 50 can be alkylated in position 8 as previously illustrated in Figure 6 to obtain compound 51.
  • This intermediate can be used to prepare 7-nitro- and 7-(hydroxyamino)benzolactams 53, 54 by peracid oxidation and reduction with zinc.
  • Diazonium chemistry can be applied to 51 to prepare the 7-iodo- and 7-mercapto derivatives 55, 56.
  • the corresponding chlorides and bromides may be obtained from 51 by action of tert-butyl nitrite and the anhydrous copper(II) halides ("Alkyl Nitrite-Metal Halide Deamination Reactions. 2. Substitutive Deamination of Arylamines by Alkyl Nitrites and Copper(II) Halides. A Direct and Remarkably Efficient Conversion of Arylamines to Aryl Halides" Doyle, M. P.; et al. J. Org. Chem. 1977, 42, 2426-2431).
  • the corresponding fluoride may be obtained from 51 using a procedure similar to that described in "A Mild and Efficient Method of Aromatic Fluorination" Rosenfeld, M. N.; Widdowson, D. A. J Chem. Soc, Chem. Commun. 1979, 914-916.
  • Compounds of formula I wherein R, and R 2 together are -
  • Iodophenol 58 and the iodoaniline 59 can be alkylated with the allylic halide 60, and the resulting intermediates cyclized under Pd catalysis to obtain the benzofuran 63 ("Synthesis of Benzofurans, Tetrahydrobenzopyrans, and Related Cyclic Ethers via Cyclic Carbopalladation” Negishi, E.; et al. Heterocycles 1989, 28, 55-58) and the indole 64 ("Conversion of 2-Halo-N-allylanilines to Indoles via Palladium(O) Oxidative Addition-Insertion Reactions" Odle, R.; et al. J. Org. Chem.
  • An intermediate of formula 28 can alternatively be prepared as illustrated in Figure 10.
  • 1 ,3-Cyclohexanedione 69 can be alkylated with aziridine 68 obtained by coupling the serine and valine building blocks. Removal ofthe N-protective group yields a compound which can close to the eight-membered lactam ring under conditions of enamine formation (high dilution), to give enaminone 73. Aromatization of enaminone 73, using standard conditions, gives compound 75, which can be protected to give an intermediate of formula 28.
  • Compounds of formula I can also be prepared using a sequence similar to that described in the previous paragraph. As shown in Figure 10, alkylation ofthe dianion of 69 gives the substituted cyclohexanedione 70.
  • salts are organic acid addition salts formed with acids which form a physiological acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate, ⁇ -ketoglutarate, and ⁇ -glycerophosphate.
  • Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts.
  • salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion.
  • a sufficiently basic compound such as an amine
  • a suitable acid affording a physiologically acceptable anion.
  • Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example calcium) salts of carboxylic acids can also be made.
  • the compounds of formula I can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes.
  • a mammalian host such as a human patient
  • the present compounds may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be inco ⁇ orated directly with the food ofthe patient's diet.
  • the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like.
  • Such compositions and preparations should contain at least 0.1% of active compound.
  • the percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% ofthe weight of a given unit dosage form.
  • the amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.
  • the tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added.
  • a liquid carrier such as a vegetable oil or a polyethylene glycol.
  • any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed.
  • the active compound may be inco ⁇ orated into sustained-release preparations and devices.
  • the active compound may also be administered intravenously or intraperitoneally by infusion or injection.
  • Solutions ofthe active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • the pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes.
  • the liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof.
  • the proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged abso ⁇ tion ofthe injectable compositions can be brought about by the use in the compositions of agents delaying abso ⁇ tion, for example, aluminum monostearate and gelatin.
  • Sterile injectable solutions are prepared by inco ⁇ orating the active compound in the required amount in the appropriate solvent with various ofthe other ingredients enumerated above, as required, followed by filter sterilization.
  • the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.
  • the present compounds may be applied in pure form, i.e., when they are liquids. However, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid.
  • Useful dosages of the compounds of formula I can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949.
  • the concentration ofthe compound(s) of formula I in a liquid composition will be from about 0.1-25 wt-%, preferably from about 0.5-10 wt-%.
  • the concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5 wt-%, preferably about 0.5-2.5 wt- %.
  • Single dosages for injection, infusion or ingestion will generally vary between 50-1500 mg, and may be administered, i.e., 1-3 times daily, to yield levels of about 0.5 - 50 mg/kg, for adults.
  • the invention includes a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of formula I as described hereinabove; or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable diluent or carrier.
  • the isozyme selectivity of representative compounds of the invention and ILV (3) was determined by investigating their ability to displace [ 3 H]PDBU binding to recombinant PKC isozymes expressed in the baculovirus system, as described by Kazanietz, M. G.; et al. Characterization of ligand and substrate specificity for the calcium-dependent and calcium-independent PKC isozymes. Mol. Pharmacol. 1993, 44, 298-307.
  • the compound of Example 1 (17) showed a higher affinity for the ⁇ and ⁇ isozymes, in comparison to ⁇ , ⁇ , and ⁇ , with approximately a ten-fold difference in affinity between PKC ⁇ and ⁇ . It is apparent that the ectronic effect of the acetylenic group influences isozyme selectivity, since the saturated alkyl compound of Example 2 (18) shows only a four-fold difference in the K ( for ⁇ versus ⁇ .
  • Table 1- K values ⁇ SEM for the inhibition of [ ⁇ ]PBDU binding by the compounds tested.
  • Representative compounds of the invention were also tested for antiproliferafive activity against breast carcinoma cell lines MCF-7 and MDA- MB-231 (Yu, G.; et al. Transfection with protein kinase C ⁇ confers increased multidrug resistance to MCF-7 cells expressing P-glycoprotein. Cancer Commun. 1991, 3, 181-189). Exposure of MCF-7 and MDA-MB-231 cells to the compound of Example 1 (17) for four days resulted in IC 50 values of 20 and 30 ⁇ M, respectively, whereas ILV (3) was inactive ( Figure 13).
  • PKC isozyme levels were determined in MCF-7 and MDA-MB- 231 cells exposed to the compound of Example 1 (17) for 24 hours ( Figure 14).
  • MCF-7 cells PKC ⁇ and PKC ⁇ were virtually eliminated, PKC ⁇ was reduced to a lesser extent, and PKC ⁇ was unchanged.
  • MDA-MB-231 cells exhibited a similar reduction in PKC ⁇ , whereas PKC ⁇ and ⁇ were slightly reduced, while PKC ⁇ and PKC ⁇ remained unchanged.
  • the invention includes a method for modulating PKC in a mammal comprising administering to said mammal a pharmaceutically effective dose of a compound of formula I; or a pharmaceutically acceptable salt thereof.
  • the invention also includes a method for the treatment of cancer in a mammal comprising administering to said mammal a pharmaceutically effective dose of a compound of formula I; or a pharmaceutically acceptable salt thereof.
  • Compounds of formula (I) are generally effective to treat mammalian conditions associated with pathological cellular proliferation. In addition to the utilities described above, they may also be useful to treat conditions which include restenosis, atherosclerosis, coronary heart disease, thrombosis, myocardial infarction, stroke, uterine fibroid or fibroma, and obliterative disease of vascular grafts and transplanted organs.
  • the invention will now be illustrated by way ofthe following non-limiting Examples.
  • the intermediate 16 was prepared as follows ( Figure 1 1). a. (5)-0-Acetyl-2-[(ethoxycarbonyl)amino]-3-phenyl-l-propanol ( Figure 1 1).
  • this product was directly 5 dissolved in 10 mL of dried CH 2 C1 2 and the solution was cooled to 0 °C before 10 mL of CF 3 COOH was added. The mixture was stirred at 0 °C for 2 hours, and the volatiles were removed in vacuo below 30 °C. The residue was dissolved in 100 mL of EtOAc, and 5 mL of saturated aqueous NaHCO, solution was added. The mixture was heated at 80 °C for 24 hours with vigorous stirring. After 0 cooling to room temperature, 40 mL of water was added. The organic layer was separated, and the aqueous layer was extracted with EtOAc (4 x 20 mL).
  • this product was directly dissolved in 10 mL of dried methylene chloride, and the resulting solution was cooled at 0 °C prior to the addition of 10 mL of trifluoroacetic acid.
  • the reaction mixture was stirred at 0 °C for 2 hours, and the trifluoroactic acid was removed in vacuo at below 30 °C.
  • the residue was dissolved in 100 mL of ethyl acetate and 10 mL of a saturated aqueous NaHCO 3 solution. The mixture was heated at 80 °C for 24 hours with vigorous stirring.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Composés de la formule (I) et leurs sels pharmaceutiquement acceptable. Dans ladite formule, R1 est H, alkyle(C1-C5), OR?a, SRa, N(Ra)(Rb¿), halo, NO¿2?, NHC(O)[(C1-C4)alkyle] ou NHOH; R2 est un groupe (C5-C22) hydrocarbyle comprenant éventuellement 1-3 liaisons doubles, 1-2 liaisons triples ou un mélange de ces deux configurations, ou un aryl(C6-C12))alkyle (C2-C10), la fraction alkyle comprenant éventuellement 1-2 liaisons doubles, 1-2 liaisons triples ou un mélange de ces configurations; ce groupe hydrocarbyle (C5-C22) ou ce groupe aryl(C6-C12)alkyle (C2-C10) pouvant éventuellement être substitué par 1 ou 2 substituts choisis indépendamment dans le groupe comprenant halo, hydroxy, cyano, nitro, alkyle (C1-C5), alcoxy (C1-C5), trifluorométhyle, trifluorométhoxy, -C(=O)O alkyle (C1-C5) et N(R?e)(Rf); R¿1 et R2 ensemble constituent -CH(Rc)-CH2-C(O)-N(Rd)-, -C(Rc)=CH-C(O)N(Rd)-, -C(Rc)=CH-N(Rd)- ou -C(Rc)=CH-O-; R¿3? est H, OH ou halo; R?a et Rb¿ sont indépendamment H ou alkyle (C¿1?-C5); R?c¿ est un groupe hydrocarbyle (C¿5?-C22); R?d¿ est H ou alkyle (C¿1?-C5); R?e et Rf¿ sont indépendamment hydrogène, alkyle (C¿1?-C5) ou alcanoyle (C1-C5), ou constituent avec l'azote auquel ils sont fixés pyrrolidino, pipéridino ou morpholino; Z est H ou alkyle (C1-C5); et Y est H ou alkyle (C1-C5). Ces substances sont des modulateurs de PKC et sont utiles au traitement, notamment, du cancer chez les mammifères. Sont également décrits des compositions pharmaceutiques constituées de composés de la formule (I), des procédés de préparation de composés de la formule (I), et des intermédiaires utiles dans la préparation de composés de formule (I).
PCT/US1997/008141 1996-05-10 1997-05-09 Derives de benzodizocine a substitution 8-hydrocarbyle, leur preparation et leur utilisation comme modulateurs de la proteine kinase c (pkc) WO1997043268A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP97925564A EP0918757A1 (fr) 1996-05-10 1997-05-09 Derives de benzodizocine a substitution 8-hydrocarbyle, leur preparation et leur utilisation comme modulateurs de la proteine kinase c (pkc)
AU30666/97A AU3066697A (en) 1996-05-10 1997-05-09 8-hydrocarbyl substituted benzodizocine derivatives, their preparation and the ir use as protein kinase c (=pkc) modulators
JP09541093A JP2000510149A (ja) 1996-05-10 1997-05-09 ８―炭化水素基置換化ベンゾジゾシン誘導体、その製造及びタンパク質キナーゼｃ（＝ｐｋｃ）調節因子としてのその利用

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US1753296P 1996-05-10 1996-05-10
US60/017,532 1996-05-10

Publications (1)

Publication Number Publication Date
WO1997043268A1 true WO1997043268A1 (fr) 1997-11-20

Family

ID=21783129

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1997/008141 WO1997043268A1 (fr) 1996-05-10 1997-05-09 Derives de benzodizocine a substitution 8-hydrocarbyle, leur preparation et leur utilisation comme modulateurs de la proteine kinase c (pkc)

Country Status (5)

Country Link
EP (1) EP0918757A1 (fr)
JP (1) JP2000510149A (fr)
AU (1) AU3066697A (fr)
CA (1) CA2253463A1 (fr)
WO (1) WO1997043268A1 (fr)

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7057046B2 (en) 2002-05-20 2006-06-06 Bristol-Myers Squibb Company Lactam glycogen phosphorylase inhibitors and method of use
US7098235B2 (en) 2002-11-14 2006-08-29 Bristol-Myers Squibb Co. Triglyceride and triglyceride-like prodrugs of glycogen phosphorylase inhibiting compounds
WO2008100450A2 (fr) 2007-02-09 2008-08-21 Blanchette Rockefeller Neurosciences Institute Effets thérapeutiques de bryostatines, de bryologues et d'autres substances apparentées sur l'altération de la mémoire induite par une ischémie/un accident vasculaire cérébral et une lésion cérébrale
EP1961447A2 (fr) 2004-05-18 2008-08-27 Blanchette Rockefeller Neurosciences Institute traitement de troubles depressifs avec des activateurs de la pkc
EP2172246A2 (fr) 2002-03-07 2010-04-07 Blanchette Rockefeller Neurosciences Institute Méthodes pour le traitement de la maladie d'Alzheimer et l'amélioration cognitive
WO2012006525A2 (fr) 2010-07-08 2012-01-12 Alkon Daniel L Activateurs de pkc et anticoagulant dans un régime pour le traitement d'un accident vasculaire cérébral
US8703812B2 (en) 2005-07-29 2014-04-22 Blanchette Rockefeller Neurosciences Institute Protein synthesis required for long-term memory is induced by PKC activation on days preceding associative learning
EP2754448A2 (fr) 2007-02-09 2014-07-16 Blanchette Rockefeller Neurosciences, Institute Effets thérapeutiques des bryostatines, de leurs analogues, et d'autres substances connexes sur l'affaiblissement de la mémoire induite par un traumatisme crânien et sur les lésions cérébrales
EP2762136A1 (fr) 2006-07-28 2014-08-06 Blanchette Rockefeller Neurosciences, Institute Procédés de stimulation de la croissance cellulaire, remodelage synaptique et consolidation de la mémoire à long terme
WO2014145316A1 (fr) 2013-03-15 2014-09-18 Alkon Daniel L Méthodes d'identification d'activateurs pkc neuroprotecteurs
US9066923B2 (en) 2002-03-07 2015-06-30 Blanchette Rockefeller Neurosciences Institute Methods for Alzheimer's disease treatment and cognitive enhancement
WO2015148975A1 (fr) 2014-03-27 2015-10-01 Alkon Daniel L Compositions et méthodes pour traiter une maladie de niemann-pick disease
WO2017099591A1 (fr) 2015-12-07 2017-06-15 Stichting Het Nederlands Kanker Instituut-Antoni van Leeuwenhoek Ziekenhuis Traitement de cancers à mutation braf résistants aux inhibiteurs

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995009160A1 (fr) * 1993-09-30 1995-04-06 Shionogi & Co., Ltd. Derive de benzolactame
WO1996040614A1 (fr) * 1995-06-07 1996-12-19 Procyon Pharmaceuticals, Inc. Modulateurs .x. de proteine kinase c

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1995009160A1 (fr) * 1993-09-30 1995-04-06 Shionogi & Co., Ltd. Derive de benzolactame
EP0721945A1 (fr) * 1993-09-30 1996-07-17 Shionogi & Co., Ltd. Derive de benzolactame
WO1996040614A1 (fr) * 1995-06-07 1996-12-19 Procyon Pharmaceuticals, Inc. Modulateurs .x. de proteine kinase c

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ENDO Y. ET AL.: "Role of the hydrophobic moiety of tumor promoters. Synthesis and activity of 9-alkylated benzolactams", CHEMICAL & PHARMACEUTICAL BULLETIN (JAPAN), vol. 44, no. 5, May 1996 (1996-05-01), pages 1138 - 1140, XP002039504 *
ENDO Y. ET AL.: "Role of the hydrophobic moiety of tumor promoters. Synthesis and activity of benzolactams with alkyl substituents at various positions", CHEMICAL & PHARMACEUTICAL BULLETIN (JAPAN), vol. 45, no. 2, February 1997 (1997-02-01), pages 424 - 426, XP002039501 *
ENDO Y. ET AL.: "Synthesis, conformation, and biological activity of teleocidin mimics, benzolactams. A clarification of the conformational flexibility problem in structure-activity studies of teleocidins", JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, vol. 118, no. 8, 28 February 1996 (1996-02-28), pages 1841 - 1855, XP002039505 *
ITAI A. ET AL.: "Advanced computational docking of two teleocidin congeners to cys2 domain of protein kinase C.delta.", CHEMICAL & PHARMACEUTICAL BULLETIN (JAPAN), vol. 45, no. 3, March 1997 (1997-03-01), pages 573 - 575, XP002039502 *
KOZIKOWSKI A.P. ET AL.: "Modeling, chemistry, and biology of the benzolactam analogs of indolactam V (ILV). 2. Identification of the binding site of the benzolactams in the CRD2 activator-binding domain of PKC.delta. and discovery of an ILV analog of improved isoenzyme selectivity", JOURNAL OF MEDICINAL CHEMISTRY, vol. 40, no. 9, April 1997 (1997-04-01), pages 1316 - 1326, XP002039503 *

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9345685B2 (en) 2002-03-07 2016-05-24 Blanchette Rockefeller Neuroscience Institute Methods for Alzheimer's Disease treatment and cognitive enhancement
US9066923B2 (en) 2002-03-07 2015-06-30 Blanchette Rockefeller Neurosciences Institute Methods for Alzheimer's disease treatment and cognitive enhancement
EP2172246A2 (fr) 2002-03-07 2010-04-07 Blanchette Rockefeller Neurosciences Institute Méthodes pour le traitement de la maladie d'Alzheimer et l'amélioration cognitive
US9539235B2 (en) 2002-03-07 2017-01-10 Cognitive Research Enterprises, Inc Methods for Alzheimer's disease treatment and cognitive enhancement
US9446020B2 (en) 2002-03-07 2016-09-20 Blanchette Rockefeller Neurosciences Institute Methods for Alzheimer'S disease treatment and cognitive enhancement
US7057046B2 (en) 2002-05-20 2006-06-06 Bristol-Myers Squibb Company Lactam glycogen phosphorylase inhibitors and method of use
US7425550B2 (en) 2002-05-20 2008-09-16 Bristol-Meyers Squibb Company Lactam glycogen phosphorylase inhibitors and method of use
US7098235B2 (en) 2002-11-14 2006-08-29 Bristol-Myers Squibb Co. Triglyceride and triglyceride-like prodrugs of glycogen phosphorylase inhibiting compounds
EP1961447A2 (fr) 2004-05-18 2008-08-27 Blanchette Rockefeller Neurosciences Institute traitement de troubles depressifs avec des activateurs de la pkc
EP2570155A2 (fr) 2004-05-18 2013-03-20 Blanchette Rockefeller Neurosciences Institute Traitement des troubles dépressifs
US10010584B2 (en) 2004-05-18 2018-07-03 West Virginia University Treatment of depressive disorders
US8703812B2 (en) 2005-07-29 2014-04-22 Blanchette Rockefeller Neurosciences Institute Protein synthesis required for long-term memory is induced by PKC activation on days preceding associative learning
EP2762136A1 (fr) 2006-07-28 2014-08-06 Blanchette Rockefeller Neurosciences, Institute Procédés de stimulation de la croissance cellulaire, remodelage synaptique et consolidation de la mémoire à long terme
EP2754448A2 (fr) 2007-02-09 2014-07-16 Blanchette Rockefeller Neurosciences, Institute Effets thérapeutiques des bryostatines, de leurs analogues, et d'autres substances connexes sur l'affaiblissement de la mémoire induite par un traumatisme crânien et sur les lésions cérébrales
EP2737904A2 (fr) 2007-02-09 2014-06-04 Blanchette Rockefeller Neurosciences Institute Effets thérapeutiques de bryostatines, de bryologues et d'autres substances apparentées sur l'altération de la mémoire induite par une ischémie/un accident vasculaire cérébral et une lésion cérébrale
EP2959914A1 (fr) 2007-02-09 2015-12-30 Blanchette Rockefeller Neurosciences, Institute Effets thérapeutiques des bryologues sur les troubles de la mémoire et les lésions cérébrales suite à un traumatisme crânien
US9974832B2 (en) 2007-02-09 2018-05-22 Cognitive Research Enterprises, Inc. Therapeutic effects of bryostatins, bryologs, and other related substances on head trauma-induced memory impairment and brain injury
EP3332797A2 (fr) 2007-02-09 2018-06-13 Blanchette Rockefeller Neurosciences Institute Effets thérapeutiques des bryostatines, bryologues et autres substances apparentées sur les troubles de la mémoire et les lésions cérébrales suite à un traumatisme crânien
WO2008100450A2 (fr) 2007-02-09 2008-08-21 Blanchette Rockefeller Neurosciences Institute Effets thérapeutiques de bryostatines, de bryologues et d'autres substances apparentées sur l'altération de la mémoire induite par une ischémie/un accident vasculaire cérébral et une lésion cérébrale
WO2012006510A1 (fr) 2010-07-08 2012-01-12 Alkon Daniel L Activateurs des protéine kinases c à base d'acides gras et anticoagulant pour le traitement de l'accident vasculaire cérébral
WO2012006516A2 (fr) 2010-07-08 2012-01-12 Alkon Daniel L Activateurs de la protéine kinase c indirects et de type dag et anticoagulant pour le traitement de l'accident vasculaire cérébral
WO2012006525A2 (fr) 2010-07-08 2012-01-12 Alkon Daniel L Activateurs de pkc et anticoagulant dans un régime pour le traitement d'un accident vasculaire cérébral
WO2014145316A1 (fr) 2013-03-15 2014-09-18 Alkon Daniel L Méthodes d'identification d'activateurs pkc neuroprotecteurs
WO2015148975A1 (fr) 2014-03-27 2015-10-01 Alkon Daniel L Compositions et méthodes pour traiter une maladie de niemann-pick disease
WO2017099591A1 (fr) 2015-12-07 2017-06-15 Stichting Het Nederlands Kanker Instituut-Antoni van Leeuwenhoek Ziekenhuis Traitement de cancers à mutation braf résistants aux inhibiteurs

Also Published As

Publication number Publication date
CA2253463A1 (fr) 1997-11-20
AU3066697A (en) 1997-12-05
JP2000510149A (ja) 2000-08-08
EP0918757A1 (fr) 1999-06-02

Similar Documents

Publication Publication Date Title
US11065234B2 (en) Estrogen receptor modulators
EP0708085B1 (fr) Ethers antiviraux des isostères du substrat de l'aspartate-protéase
WO1997043268A1 (fr) Derives de benzodizocine a substitution 8-hydrocarbyle, leur preparation et leur utilisation comme modulateurs de la proteine kinase c (pkc)
DK2026775T3 (en) USE OF SYK tyrosine kinase inhibitors for treating cell proliferative DISORDERS
US7678783B2 (en) Macrocyclic tertiary amine beta-secretase inhibitors for the treatment of alzheimer's disease
EA021983B1 (ru) Производные диоксабицикло[3.2.1]октан-2,3,4-триола
US10730864B2 (en) RAD51 inhibitors and uses thereof
JPH0826036B2 (ja) 生理活性物質ｋ−２５２の誘導体
KR20200012316A (ko) 히스톤 탈아세틸화효소 6 억제제로서의 1,3,4-옥사다이아졸 유도체 화합물 및 이를 포함하는 약제학적 조성물
CN105263924A (zh) Cxcr7受体调节剂
SK119997A3 (en) New cryptophycins from synthesis
US20010034365A1 (en) Halogen substituted carbamate compounds from 2-phenyl-1,2-ethanediol
US20040018995A1 (en) Derivatives of monosaccharides as cell adhesion inhibitors
US6372775B1 (en) Reversal of multidrug resistance in human colon carcinoma cells
WO2009018344A1 (fr) Agents anticancéreux
NZ198117A (en) Fluorinated alkylene diamines and pharmaceutical compositions
OGAWA et al. Synthesis and Antiarrhythmic Activity of 2, 5-Disubstituted 2, 3-Dihydro-1, 2, 5-benzothiadiazepin-4 (5H)-one 1, 1-Dioxides
US4503058A (en) Therapeutically useful 3,7a-diazacyclohepta[j,k]fluorene derivatives
JPH0222252A (ja) ５―ヒドロキシトリプタミン―１ａ―受容体結合化合物
WO1993020695A1 (fr) Dibenz (c,e) azepines polyhydroxylees utiles comme inhibiteurs de la proteine-kinase c
US20040248877A1 (en) Polycyclic diazodioxide-based Bcl-2 protein antagonists and use thereof
WO2008137476A1 (fr) Dérivés arylthioacétamide carboxylate en tant que inhibiteurs fkbp dans le traitement des maladies neurologiques
JPH0834789A (ja) ピロロインドールカルボン酸エステル誘導体及びその製造方法
JPH0834788A (ja) ピロロベンゾカルバゾール誘導体及びその製造方法
JPH08151380A (ja) ビス（トリフルオロメチルピロロインドールカルボン酸エステル）誘導体及びその製造方法

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GE GH HU IL IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK TJ TM TR TT UA UG US UZ VN YU AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH KE LS MW SD SZ UG AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF

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
ENP Entry into the national phase

Ref document number: 2253463

Country of ref document: CA

Ref country code: CA

Ref document number: 2253463

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 1997925564

Country of ref document: EP

REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

WWP Wipo information: published in national office

Ref document number: 1997925564

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1997925564

Country of ref document: EP