WO2007054717A2 - STAUROSPORINE DERIVATIVES FOR THE TREATMENT OF LEUKEMIA CHARACTERISED BY A SPTBNl -FLT3 MUTATION - Google Patents

STAUROSPORINE DERIVATIVES FOR THE TREATMENT OF LEUKEMIA CHARACTERISED BY A SPTBNl -FLT3 MUTATION Download PDF

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WO2007054717A2
WO2007054717A2 PCT/GB2006/004210 GB2006004210W WO2007054717A2 WO 2007054717 A2 WO2007054717 A2 WO 2007054717A2 GB 2006004210 W GB2006004210 W GB 2006004210W WO 2007054717 A2 WO2007054717 A2 WO 2007054717A2
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flt3
translocation
typically
hydrogen
substituted
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PCT/GB2006/004210
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French (fr)
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WO2007054717A3 (en
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Nicholas Cross
Francis Hector Grand
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University Of Southampton
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/553—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having at least one nitrogen and one oxygen as ring hetero atoms, e.g. loxapine, staurosporine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • This invention relates generally to the analytical testing of tissue samples in vitro, and more particularly to aspects of genetic mutations and translocations of SPTBN 1 with FLT3.
  • Theranostic tests are also useful to select subjects for treatments that are particularly likely to benefit from the treatment or to provide an early and objective indication of treatment efficacy in individual subjects, so that the treatment can be altered with a minimum of delay.
  • Theranostics are useful in clinical diagnosis and management of a variety of diseases and disorders, which include, but are not limited to, e.g., cardiovascular disease, cancer, infectious diseases, Alzheimer's disease and the prediction of drug toxicity or drug resistance.
  • Theranostic tests may be developed in any suitable diagnostic testing format, which include, but is not limited to, e.g., immunoassay, immunohistochemical tests, clinical chemistry, cell-based technologies, and nucleic acid tests.
  • Spectrin is composed of alpha- and beta- subunits that form antiparallel dimers that self associate, giving rise to the cytoskeletal spectrin tetramer.
  • Beta subunits such as spectrin beta nonerythrocytic 1 ("SPTBN1" OMIM accno. 182790), contain most of the spectrin binding activity (Hayes et al., J. Cell ScL 113: 2023-2034 (2000). Disruption of SPTBN1 leads to the disruption of TGF-beta (OMIM accno. 190180) signaling by Smad proteins in mice.
  • SPTBN 1 null (-/-) mice exhibit a phenotype similar to Smad2/Smad3 double heterozygous mice, with midgestational death due to gastrointestinal, liver, neural, and heart defects.
  • TGF-beta triggers phosphorylation and association of SPTBN1 with Smad3 and Smad4, followed by nuclear translocation. Tang et al. Science 299: 574-577 (2003).
  • SPTBN1 deficiency results in mislocalization of Smad3 and Smad4 and loss of TGF-beta- dependent transcriptional response.
  • FLT3 kinase is a member of the type III receptor tyrosine kinase (RTK) family.
  • FLT3 FLT3 (fms-like tyrosine kinase) is also known as FLk-2 (fetal liver kinase 2).
  • FLk-2 fetal liver kinase 2
  • AML acute myeloid leukemia
  • AML/TMDS AML with trilineage myelodysplasia
  • ALL acute lymphoblastic leukemia
  • MDS myelodysplastic syndrome
  • the most common mutation involves an in-frame duplication within the juxtamembrane domain, with an additional 5-10% of patients having a point mutation at asparagine 835. Both of these mutations are associated with constitutive activation of the tyrosine kinase activity of FLT3, and result in proliferation and viability signals in the absence of ligand. Patients expressing the mutant form of the receptor have been shown to have a decreased chance for cure. Thus, there is accumulating evidence for a role for hyperactivated (mutated) FLT3 kinase activity in human cancers.
  • the invention provides for the use of a FLT3 modulating agent in the manufacture of a medicament for the treatment of cancer in a selected patient population.
  • the patient population is selected on the basis of the genotype of the patients SPTBN 1- FLT3 genetic translocation indicative of efficacy of the FLT3-modulating agent in treating cancer.
  • the cancer can be atypical chronic myeloid leukemia.
  • the invention further provides a method for treating cancer in a subject.
  • the genotype or haplotype of a subject is obtained to identify patients with a translocation between the SPTBN1-FLT3 gene loci, so that the genotype and/or haplotype is indicative of a propensity of the cancer to respond to the drug.
  • the anti-cancer therapy is administered to the subject.
  • FIG. 1 panels 1A-1 D provides the full length cDNA (SEQ ID NO:1) sequence of the SPTBN1-FLT3 gene fusion and its encoded 570 aa polypeptide (SEQ ID NO:2).
  • Leukemia is a progressive, malignant disease of the blood-forming organs, characterized by distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is classified as acute or chronic, which refer to the degree of cell differentiation, and as myelogenous or lymphocytic, which refer to the predominant type of cell involved.
  • Myeloid leukemia is a variety of leukemia in which some types of white blood cells, originating in the myeloid tissue of the bone marrow, proliferate and suppress healthy red and white blood cells.
  • the present invention relates to the efficacy of staurosporine derivatives in the treatment of atypical chronic myeloid leukemia containing a SPTBN1-FLT3 translocation.
  • the present invention relates to the use of staurosporine derivatives of formula A, B, C, D, I 1 II, III, IV, V, Vl and VII: (hereinafter: "STAUROSPORINE DERIVATIVES”) for the preparation of a drug for the treatment of diseases involving a SPTBN1-FLT3 translocation and associated signalling pathways, especially for the curative and/or prophylactic treatment of atypical chronic myeloid leukemia, and to a method of treating diseases involving SPTBN1-FLT3 translocation-associated signalling pathways.
  • STAUROSPORINE DERIVATIVES for the preparation of a drug for the treatment of diseases involving a SPTBN1-FLT3 translocation and associated signalling pathways, especially for the curative and/or prophylactic treatment of atypical chronic myeloid leukemia, and to a method of treating diseases involving SPTBN1-FLT3 translocation-associated signalling pathways.
  • the invention relates to the use of staurosporine derivatives of formula
  • R 5 is hydrogen, an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms in each case, or a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms in each case, and in each case up to 9 heteroatoms, or acyl with up to 30 carbon atoms;
  • X stands for 2 hydrogen atoms; for 1 hydrogen atom and hydroxy; for O; or for hydrogen and lower alkoxy;
  • Q and Q' are independently a pharmaceutically acceptable organic bone or hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N- mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N- mono- or N,N-di-substituted aminosulfonyl; or a salt thereof, if at least one salt-forming group is present, or hydrogenated derivative thereof, for the preparation of a pharmaceutical composition for the treatment of diseases involving SPTBN1-FLT3 translocation associated signalling pathways.
  • organic bone refers to a pharmacologically acceptable organic chemical structure, such as but not limited to hydrocarbyl radical or an acyl radical Ac, which radicals preferably have a maximum of 30 carbon atoms.
  • the hydrocarbyl radical is an acyclic (aliphatic), , carbocyclic or carbocyclic-acyclic hydrocarbon radical having a maximum total number of carbon atoms of preferably 30 and, especially, 18, which may be saturated or unsaturated, unsubstituted or substituted.
  • heterocyclic radical heterocyclyl radical
  • heterocyclic-acyclic radical a heterocyclic radical
  • Unsaturated radicals are those that contain one or more, especially conjugated and/or isolated, multiple bonds (double and/or triple bonds).
  • cyclic radicals also includes aromatic radicals, for example those in which at least one 6- membered carbocyclic ring or one 5- to 8-membered heterocyclic ring contains the maximum number of non-cumulated double bonds.
  • Carbocyclic radicals in which at least one ring is in the form of a 6-membered aromatic ring (that is to say a benzene ring) are referred to as aryl radicals.
  • An acyclic unsubstituted hydrocarbon radical is especially a straight or branched lower alkyl, lower alkenyl, lower alkadienyl or lower alkynyl radical.
  • the double bond is located especially in a position higher than the alpha-position to the free valency.
  • a carbocyclic hydrocarbon radical is especially a mono-, bi- or polycyclic cycloalkyl, cycloalkenyl or cycloalkadienyl radical, or a corresponding aryl radical.
  • Preferred are radicals having a maximum of 14, especially 12, ring carbon atoms and having 3- to 8- membered, preferably 5- to 7-membered, especially 6-membered, rings; they may also carry one or more, for example two, acyclic radicals, for example those mentioned above, and especially lower alkyl radicals, or other carbocyclic radicals.
  • Carbocyclic-acyclic radicals are those in which an acyclic radical, especially one having a maximum of 10, preferably a maximum of 6, carbon atoms, such as, especially, methyl, ethyl or vinyl, carries one or more of the carbocyclic, optionally aromatic radicals defined above. Mention is made especially of cycloalkyl-lower alkyl and aryl-lower alkyl radicals, and also analogues thereof unsaturated in the ring and/or chain, that carry the ring at the terminal carbon atom of the chain.
  • Linkers between the acyclic (aliphatic) or carbocyclic radicals may be selected from, but not limited to, straight or branched lower alkyl, lower alkenyl, lower alkadienyl or lower alkynyl radical, etherified or esterified hydroxy, amino, -O-, -S-, carbonyl, -NO-, -SO-, carbonyldioxy, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl.
  • leukemia refers to the clonal proliferation in bone marrow or blood of one or more of the haematopoietic lineages (World Health Organization Classification of Tumours, 2001).
  • Myeloid leukemia is the clonal proliferation of one or more of the myeloid lineages.
  • Chronic myeloid leukemia (“CML”) is a myeloproliferative disease that originates in an abnormal bone marrow stem cell and is consistently associated with the Philadelphia (Ph) chromosome and/or the BCR-ABL fusion gene.
  • the disease is bi- or triphasic: an initial indolent chronic phase (CP-CML) is followed by one or both of the aggressive transformed stages, accelerated phase (CML-AP) or blast phase (CML-BP).
  • CML-AP accelerated phase
  • CML-BP blast phase
  • aCML is a clonal leukemic disorder with myeloproliferative features like CML that does not have the Philadelphia chromosome or the BCR-ABL fusion gene.
  • leukemia means preferably atypical chronic myeloid leukemia.
  • the invention relates in particular to the use of staurosporines derivatives of formula
  • Z stands for hydrogen or lower alkyl; and either the two bonds characterised by wavy lines are absent in ring A and replaced by 4 hydrogen atoms, and the two wavy lines in ring B each, together with the respective parallel bond, signify a double bond; or the two bonds characterised by wavy lines are absent in ring B and replaced by a total of
  • Lower alkyl is especially methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, and also pentyl, hexyl, or heptyl.
  • Unsubstituted or substituted alkyl is preferably Ci-C 20 alkyl, especially lower alkyl, typically methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, which is unsubstituted or substituted especially by halogen, such as fluorine, chlorine, bromine, or iodine, C 6 -C 14 aryl, such as phenyl or naphthyl, hydroxy, etherified hydroxy, such as lower alkoxy, phenyl-lower alkoxy or phenyloxy, esterified hydroxy, such as lower alkanoyloxy or benzoyloxy, amino, mono- or disubstituted amino, such as lower alkylamino, lower alkanoylamino, phenyl-lower alkylamino, N, N-di-lower alkylamino
  • Halogen is preferably fluorine, chlorine, bromine, or iodine, especially fluorine or chlorine.
  • Etherified hydroxy is especially lower alkoxy, C 6 -C 14 aryloxy, such as phenyloxy, or C 6 -C 14 aryl-lower alkoxy, such as benzyloxy.
  • Esterified hydroxy is preferably lower alkanoyloxy or C 6 -C 14 arylcarbonyloxy, such as benzoyloxy.
  • Mono- or disubstituted amino is especially amino monosubstituted or disubstituted by lower alkyl, C e -Ci 4 aryl-lower alkyl, lower alkanoyl, or C 6 -Ci 2 arylcarbonyl • )
  • Substituted mercapto is especially lower alkylthio, C 6 -C 14 arylthio, C 6 -C 14 aryl- lower alkylthio, lower alkanoylthio, or C 6 -Ci 4 aryl-lower alkanoylthio.
  • Esterified carboxy is especially lower alkoxycarbonyl, C 6 -C 14 aryl-lower alkoxy- carbonyl or
  • N-Mono- or N,N-disubstituted carbamoyl is especially carbamoyl N- monosubstituted or N,N-disubstituted by lower alkyl, C 6 -C 14 aryl or C 6 -C 14 aryl-lower alkyl.
  • Substituted sulfonyl is especially C 6 -Ci 4 arylsulfonyl, such as toluenesulfonyl,
  • N-Mono- or N,N-disubstituted aminosulfonyl is especially aminosulfonyl N- monosubstituted or N,N-disubstituted by lower alkyl, C 6 -C 14 aryl or C 6 -Ci 4 aryl-lower alkyl.
  • C 6 -C 14 Aryl is an aryl radical with 6 to 14 carbon atoms in the ring system, such as phenyl, naphthyl, fluorenyl, or indenyl, which is unsubstituted or is substituted especially by halogen, such as fluorine, chlorine, bromine, or iodine, phenyl or naphthyl, hydroxy, lower alkoxy, phenyl-lower alkoxy, phenyloxy, lower alkanoyloxy, benzoyloxy, amino, lower alkylamino, lower alkanoylamino, phenyl-lower alkylamino, N,N-di-lower alkylamino, N,N-di- (phenyl-lower alkyl)amino, cyano, mercapto, lower alkylthio, carboxy, lower alkoxycarbonyl, carbamoyl, N-lower alkylcarbamo
  • indices n and m are in each case preferably 1 , 2 or especially 0.
  • compounds of formula I in which n and m are in each case 0 (zero) are especially preferred.
  • An aliphatic carbohydrate radical with up to 29 carbon atoms R 3 , R 4 ,. R 8 or Ri 0 which is substituted by acyclic substituents and preferably has a maximum of 18, especially a maximum of 12, and as a rule not more than 7 carbon atoms, may be saturated or unsaturated and is especially an unsubstituted or a straight-chain or branched lower alkyl, lower alkenyl, lower alkadienyl, or lower alkinyl radical substituted by acyclic substituents.
  • Lower alkyl is, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, and also n-pentyl, isopentyl, n-hexyl, isohexyl and n-heptyl; lower alkenyl is, for example, allyl, propenyl, isopropenyl, 2- or 3-methallyl and 2- or 3-butenyl; lower alkadienyl is, for example, 1-penta-2,4-dienyl; lower alkinyl is, for example, propargyl or 2-butinyl.
  • the double bond is especially located in a position higher than the ⁇ -position in relation to the free valency.
  • Substituents are especially the acyl radicals defined hereinbelow as substituents of R 0 , preferably free or esterified carboxy, such as carboxy or lower alkoxycarbonyl, cyano or di-lower alkylamino.
  • a carbocyclic or carbocyclic-aliphatic radical R 3 , R 4 , R 8 or R 10 with up to 29 carbon atoms in each case is especially an aromatic, a cycloaliphatic, a cycloaliphatic- aliphatic, or an aromatic-aliphatic radical which is either present in unsubstituted form or substituted by radicals referred to hereinbelow as substituents of R 0 .
  • aromatic radical (aryl radical) R 3 or R 4 is most especially a phenyl, also a naphthyl, such as 1- or 2-naphthyl, a biphenylyl, such as especially 4-biphenylyl, and also an anthryl, fluorenyl and azulenyl, as well as their aromatic analogues with one or more saturated rings, which is either present in unsubstituted form or substituted by radicals referred to hereinbelow as substituents of R 0 .
  • Preferred aromatic-aliphatic radicals are aryl-lower alkyl- and aryl-lower alkenyl radicals, e.g.
  • phenyl-lower alkyl or phenyl-lower alkenyl with a terminal phenyl radical such as for example benzyl, phenethyl, 1-, 2-, or 3-phenylpropyl, diphenylmethyl (benzhydryl), trityl, and cinnamyl, and also 1- or 2-naphthylmethyl.
  • aryl radicals carrying acyclic radicals such as lower alkyl, special mention is made of o-, m- and p_-tolyl and xylyl radicals with variously situated methyl radicals.
  • a cycloaliphatic radical R 3 , R 4 , R 8 or R 10 with up to 29 carbon atoms is especially a substituted or preferably unsubstituted mono-, bi-, or polycyclic cycloalkyl-, cycloalkenyl-, or cycloalkadienyl radical.
  • Preference is for radicals with a maximum of 14, especially 12, ring-carbon atoms and 3- to 8-, preferably 5- to 7-, and most especially 6- member rings which can also carry one or more, for example two, aliphatic hydrocarbon radicals, for example those named above, especially the lower alkyl radicals, or other cycloaliphatic radicals.
  • Preferred substituents are the acyclic substituents named hereinbelow for R 0 .
  • a cycloaliphatic-aliphatic radical R 3 , R 4 , R 8 or Ri 0 with up to 29 carbon atoms is a radical in which an acyclic radical, especially one with a maximum of 7, preferably a maximum of 4 carbon atoms, such as especially methyl, ethyl, and vinyl, carries one or more cycloaliphatic radicals as defined hereinabove.
  • an acyclic radical especially one with a maximum of 7, preferably a maximum of 4 carbon atoms, such as especially methyl, ethyl, and vinyl
  • Preferred substituents are the acyclic substituents named herein below for R 0 .
  • Heterocyclic radicals R 3 , R 4 , R 8 or R 10 with up to 20 carbon atoms each and up to 9 heteroatoms each are especially monocyclic, but also bi- or polycyclic, aza-, thia-, oxa-, thiaza-, oxaza-, diaza-, triaza-, or tetrazacyclic radicals of an aromatic character, as well as corresponding heterocyclic radicals of this type which are partly or most especially wholly saturated, these radicals - if need be - possibly carrying further acyclic, carbocyclic, or heterocyclic radicals and/or possibly mono-, di-, or polysubstituted by functional groups, preferably those named hereinabove as substituents of aliphatic hydrocarbon radicals.
  • pyrryl for example 2-pyrryl or 3-pyrryl
  • pyridyl for example 2-, 3-, or 4-pyridyl
  • thienyl for example 2- or 3-thienyl
  • furyl for example 2-furyl
  • analogous bicyclic radicals with an oxygen, sulfur, or nitrogen atom are, for example, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, or benzothienyl, typically 2- or 3- benzothienyl
  • preferred monocyclic and bicyclic radicals with several heteroatoms are, for example, imidazolyl, typically 2-pyrryl or 3-pyrryl
  • pyridyl for example 2-, 3-, or 4-pyridyl
  • thienyl for example 2- or 3-thienyl
  • furyl for example 2-fury
  • radicals may also be considered, such as 2-tetrahydrofuryl, 2- or 3-pyrrolidinyl, 2-, 3-, or 4-piperidyl, and also 2-or 3-morpholinyl, 2- or 3-thiomorpholinyl, 2-piperazinyl and N-mono- or N,N'-bis-lower alkyl-2-piperazinyl radicals.
  • These radicals may also carry one or more acyclic, carbocyclic, or heterocyclic radicals, especially those mentioned hereinabove.
  • the free valency of the heterocyclic radicals R 3 or R 4 must emanate from one of their carbon atoms.
  • Heterocyclyl may be unsubstituted or substituted by one or more, preferably one or two, of the substituents named hereinbelow for R 0 .
  • Heterocyclic-aliphatic radicals R 3 , R 4 , R 8 or R 10 especially lower alkyl radicals, especially with a maximum of 7, preferably a maximum of 4 carbon atoms, for example those named hereinabove, which carry one, two, or more heterocyclic radicals, for example those named in the preceding paragraph, the heterocyclic ring possibly being linked to the aliphatic chain also by one of its nitrogen atoms.
  • a preferred heterocyclic-aliphatic radical R-i is, for example, imidazol-1-ylmethyl, 4-methylpiperazin-1 ⁇ ylmethyl, piperazin-1-ylmethyl, 2- (morpholin-4-yl)ethyl and also pyrid-3-ylmethyl.
  • Heterocyclyl may be unsubstituted or substituted by one or more, preferably one or two, of the substituents named hereinbelow for R 0 .
  • a heteroaliphatic radical R 3 , R 4 , R 8 or Ri 0 with up to 20 carbon atoms each and up to 10 heteroatoms each is an aliphatic radical which, instead of one, two, or more carbon atoms, contains identical or different heteroatoms, such as especially oxygen, sulfur, and nitrogen.
  • R 3 , R 4 , R 8 or Ri 0 apart from acyl, is lower alkyl, particlularly methyl or ethyl; lower alkoxycarbonyl-lower alkyl, especially methoxycarbonylmethyl or 2-(tert-butoxycarbonyl)ethyl; carboxy-lower alkyl, especially carboxymethyl or 2-carboxyethyl; or cyano-lower alkyl, especially 2-cyanoethyl.
  • An acyl radical R 3 , R 4 , R 6 , R 7 , Rs, R9, or R 10 with up to 30 carbon atoms derives from a carboxylic acid, functionally modified if need be, an organic sulfonic acid, or a phosphoric acid, such as pyro- or orthophosphoric acid, esterified if need be.
  • the hydrocarbyl (hydrocarbon radical) R 0 is an acyclic (aliphatic), carbocyclic, or carbocyclic-acyclic hydrocarbon radical, with up to 29 carbon atoms each, especially up to 18, and preferably up to 12 carbon atoms, and is saturated or unsaturated, unsubstituted or substituted. Instead of one, two, or more carbon atoms, it may contain identical or different heteroatoms, such as especially oxygen, sulfur, and nitrogen in the acyclic and/or cyclic part; in the latter case, it is described as a heterocyclic radical (heterocyclyl radical) or a hetero- cyclic-acyclic radical.
  • Unsaturated radicals are those, which contain one or more, especially conjugated and/or isolated, multiple bonds (double or triple bonds).
  • cyclic radicals includes also aromatic and non-aromatic radicals with conjugated double bonds, for example those wherein at least one 6-member carbocyclic or a 5- to 8-member heterocyclic ring contains the maximum number of non-cumulative double bonds.
  • Carbocyclic radicals, wherein at least one ring is present as a 6-member aromatic ring (i.e. a benzene ring), are defined as aryl radicals.
  • An acyclic unsubstituted hydrocarbon radical R 0 is especially a straight- chained or branched lower alkyl-, lower alkenyl-, lower alkadienyl-, or lower alkinyl radical.
  • Lower alkyl R 0 is, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, and also n-pentyl, isopentyl, n-hexyl, isohexyl and n-heptyl; lower alkenyl is, for example, allyl, propenyl, isopropenyl, 2- or 3-methallyl and 2- or 3-butenyl; lower alkadienyl is, for example, 1-penta-2,4-dienyl; lower alkinyl is, for example, propargyl or 2-butinyl.
  • the double bond is especially located in a position higher than the ⁇ -position in relation to the free valency.
  • a carbocyclic hydrocarbon radical R 0 is especially a mono-, bi-, or polycyclic cycloalkyl-, cycloalkenyl-, or cycloalkadienyl radical, or a corresponding aryl radical.
  • Preference is for radicals with a maximum of 14, especially 12, ring-carbon atoms and 3- to 8-, preferably 5- to 7-, and most especially 6-member rings which can also carry one or more, for example two, acyclic radicals, for example those named above, especially the lower alkyl radicals, or other carbocyclic radicals.
  • Carbocyclic-acyclic radicals are those in which an acyclic radical, especially one with a maximum of 7, preferably a maximum of 4 carbon atoms, such as especially methyl, ethyl and vinyl, carries one or more carbocyclic, if need be aromatic radicals of the above definition. Special mention is made of cycloalkyl- lower and aryl-lower alkyl radicals, as well as their analogues which are unsaturated in the ring and/or chain, and which carry the ring at the terminal carbon atom of the chain.
  • Cycloalkyl R 0 has most especially from 3 up to and including 10 carbon atoms and is, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, as well as bicyclo[2,2,2]octyl, 2-bicyclo[2,2,1]heptyl, and adamantyl, which may also be substituted by 1 , 2, or more, for example lower, alkyl radicals, especially methyl radicals; cycloalkenyl is for example one of the monocyclic cycloalkyl radicals already named which carries a double bond in the 1-, 2-, or 3 position.
  • Cycloalkyl-lower alkyl or -lower alkenyl is for example a -methyl, -1- or -2-ethyl, -1- or -2-vinyl, -1-, -2-, or -3-propyl or -allyl substituted by one of the above-named cycloalkyl radicals, those substituted at the end of the linear chain being preferred.
  • An aryl radical R 0 is most especially a phenyl, also a naphthyl, such as 1- or
  • 2-naphthyl a biphenylyl, such as especially 4-biphenylyl, and also an anthryl, fluorenyl and azulenyl, as well as their aromatic analogues with one or more saturated rings.
  • Preferred aryl-lower alkyl and -lower alkenyl radicals are, for example, phenyl-lower alkyl or phenyl- lower alkenyl with a terminal phenyl radical, such as for example benzyl, phenethyl, 1-, 2-, or 3-phenylpropyl, diphenylmethyl (benzhydryl), trityl, and cinnamyl, and also 1- or 2-naphthyl- methyl.
  • Aryl may be unsubstituted or substituted.
  • Heterocyclic radicals including heterocyclic-acyclic radicals, are especially monocyclic, but also bi- or polycyclic, aza-, thia-, oxa-, thiaza-, oxaza-, diaza-, triaza-, or tetrazacyclic radicals of an aromatic character, as well as corresponding heterocyclic radicals of this type which are partly or most especially wholly saturated; if need be, for example as in the case of the above-mentioned carbocyclic or aryl radicals, these radicals may carry further acyclic, carbocyclic, or heterocyclic radicals and/or may be mono-, di-, or polysubstituted by functional groups.
  • heterocyclic-acyclic radicals has for example the meaning indicated for the corresponding carbocyclic-acyclic radicals.
  • they are unsubstituted or substituted monocyclic radicals with a nitrogen, oxygen, or sulfur atom, such as 2-aziridinyl, and especially aromatic radicals of this type, such as pyrrolyl, for example 2-pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3-, or 4-pyridyl, and also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl; analogous bicyclic radicals with an oxygen, sulfur, or nitrogen atom are, for example, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, or benzothi
  • radicals may also be considered, such as 2-tetrahydrofuryl, 4-tetrahydrofuryl, 2- or 3-pyrrolidyl, 2-, 3-, or 4-piperidyl, and also 2-or 3-morpholinyl, 2- or 3-thiomorpholinyl, 2- piperazinyl, and N,N'-bis-lower alkyl-2-piperazinyl radicals.
  • These radicals may also carry one or more acyclic, carbocyclic, or heterocyclic radicals, especially those mentioned hereinabove.
  • Heterocyclic-acyclic radicals are especially derived from acyclic radicals with a maximum of 7, preferably a maximum of 4 carbon atoms, for example those named hereinabove, and may carry one, two, or more heterocyclic radicals, for example those named hereinabove, the ring possibly being linked to the aliphatic chain also by one of its nitrogen atoms.
  • a hydrocarbyl may be substituted by one, two, or more identical or different substituents (functional groups); one or more of the following substituents may be considered: lower alkyl; free, etherified and esterified hydroxyl groups; carboxy groups and esterified carboxy groups; mercapto- and lower alkylthio- and, if need be, substituted phenylthio groups; halogen atoms, typically chlorine and fluorine, but also bromine and iodine; halogen-lower alky) groups; oxo groups which are present in the form of formyl (i.e.
  • aldehydo aldehydo
  • keto groups also as corresponding acetals or ketals; azido groups; nitro groups; cyano groups; primary, secondary and preferably tertiary amino groups, amino-lower alkyl, mono- or disubstituted amino-lower alkyl, primary or secondary amino groups protected by conventional protecting groups (especially lower alkoxycarbonyl, typically tert-butoxycarbony!) lower alkylenedioxy, and also free or functionally modified sulfo groups, typically sulfamoyl or sulfo groups present in free form or as salts.
  • the hydrocarbyl radical may also carry carbamoyl, ureido, or guanidino groups, which are free or which carry one or two substituents, and cyano groups.
  • groups are taken to imply also an individual group.
  • Halogen-lower alkyl contains preferably 1 to 3 halogen atoms; preferred is trifluoromethyl or chloromethyl.
  • An etherified hydroxyl group present in the hydrocarbyl as substituent is, for example, a lower alkoxy group, typically the methoxy-, ethoxy-, propoxy-, isopropoxy-, butoxy-, and tert-butoxy group, which may also be substituted, especially by (i) heterocyclyl, whereby heterocyclyl can have preferably 4 to 12 ring atoms, may be unsaturated, or partially or wholly saturated, is mono- or bicyclic, and may contain up to three heteroatoms selected from nitrogen, oxygen, and sulfur, and is most especially pyrrolyl, for example 2- pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, and also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl,
  • Such etherified hydroxyl groups are also unsubstituted or substituted phenoxy radicals and phenyl-lower alkoxy radicals, such as especially benzyloxy, benzhydryloxy, and triphenylmethoxy (trityloxy), as well as heterocyclyloxy radicals, wherein heterocyclyl can have preferably 4 to 12 ring atoms, may be unsaturated, or partially or wholly saturated, is mono- or bicyclic, and may contain up to three heteroatoms selected from nitrogen, oxygen, and sulfur, and is most especially pyrrolyl, for example 2-pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, and also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinoly
  • Etherified hydroxyl groups in this context are taken to include silylated hydroxy! groups, typically for example tri-lower alkylsilyloxy, typically tri methyls ilyloxy and dimethyl-tert-butylsilyloxy, or phenyldi-lower alkylsilyloxy and lower alkyl-diphenylsilyloxy.
  • An esterified hydroxyl group present in the hydrocarbyl as a substituent is, for example, lower alkanoyloxy.
  • a carboxyl group present in the hydrocarbyl as a substituent is one in which the hydrogen atom is replaced by one of the hydrogen radicals characterised hereinabove, preferably a lower alkyl- or phenyl-lower alkyl radical; an example of an esterified carboxyl group is lower alkoxycarbonyl or phenyl-lower alkoxycarbonyl substituted if need be in the phenyl part, especially the methoxy, ethoxy, tert-butoxy, and benzyloxycarbonyl group, as well as a lactonised carboxyl group.
  • a primary amino group -NH 2 as substituent of the hydrocarbyls may also be present in a form protected by a conventional protecting group.
  • a secondary amino group carries, instead of one of the two hydrogen atoms, a hydrocarbyl radical, preferably an unsubstituted one, typically one of the above-named, especially lower alkyl, and may also be present in protected form.
  • a tertiary amino group present in the hydrocarbyl as substituent carries 2 different or, preferably, identical hydrocarbyl radicals (including the heterocyclic radicals), such as the unsubstituted hydrocarbyl radicals characterised hereinabove, especially lower alkyl.
  • a preferred amino group is one with the formula R 1I (Ri 2 )N-, wherein R 11 and
  • R 12 are independently in each case hydrogen, unsubstituted acyclic C r C 7 -hydrocarbyl (such as especially C-i-C 4 alkyl or C 2 -C 4 alkenyl) or monocyclic aryl, aralkyl, or aralkenyl, substituted if necessary by C r C 4 -alkyl, C r C 4 -alkoxy, halogen, and/or nitro, and having a maximum of 10 carbon atoms, where the carbon-containing radicals may be interlinked through a carbon- carbon bond or an oxygen atom, a sulfur atom, or a nitrogen atom substituted if necessary by hydrocarbyl.
  • acyclic C r C 7 -hydrocarbyl such as especially C-i-C 4 alkyl or C 2 -C 4 alkenyl
  • monocyclic aryl, aralkyl, or aralkenyl substituted if necessary by C r C 4 -alkyl
  • di-lower alkylamino typically dimethylamino or diethylamino, pyrrolidino, imidazol-1-yl, piperidino, piperazino, 4-lower alkylpiperazino, morpholino, thiomorpholino and piperazino or 4-methylpiperazino, as well as diphenylamino and dibenzylamino substituted if need be, especially in the phenyl part, for example by lower- alkyl, lower-alkoxy, halogen, and/or nitro; of the protected groups, especially lower alkoxy- carbonylamino, typically tert-butoxycarbonylamino, phenyl-lower alkoxycarbonylamino, typically 4-methoxybenzyloxycarbonylamino, and 9-fluorenyl
  • Mono- or disubstituted- amino-lower alkyl is amino-lower alkyl substituted by one or two radicals, wherein amino-lower alkyl is most especially substituted by amino in the 1 -position of the lower alkyl chain and is especially aminomethyl; the amino substituents here are preferably (if 2 substituents are present in the respective amino group independently of one another) from the group comprising lower alkyl, such as especially methyl, ethyl or n-propyl, hydroxy-lower alkyl, typically 2-hydroxyethyl, C 3 -C 8 cycloalkyl, especially cyclohexyl, amino-lower alkyl, typically 3-aminopropyl or 4-aminobutyl, N-mono- or N,N-di(lower alkyl)-amino-lower alkyl, typically 3-(N,N-dimethylamino)propyl, amino, N-mono- or N,N-di
  • Disubstituted amino-lower alkyl is also a 5 or 6-membered, saturated or unsaturated heterocyclyl bonded to lower alkyl via a nitrogen atom (preferably in the 1- position) and having 0 to 2, especially 0 or 1 , other heteroatoms selected from oxygen, nitrogen, and sulfur, which is unsubstituted or substituted, especially by one or two radicals from the group comprising lower alkyl, typically methyl, and also oxo.
  • Preferred here is pyrrolidino (1-pyrrolidinyl), piperidino (1-piperidinyl), piperazino (1-piperazinyl), 4-lower alkylpiperazino, typically 4-methylpiperazino, imidazolino (1-imidazolyl), morpholino (4-morpho- linyl), or also thiomorpholino, S-oxo-thiomorpholino, or S,S ⁇ dioxothiomorpholino.
  • Lower alkylenedioxy is especially methylenedioxy.
  • a carbamoyl group carrying one or two substituents is especially amino- carbonyl (carbamoyl) which is substitiuted by one or two radicals at the nitrogen; the amino substituents here are preferably (if 2 substituents are present in the respective amino group independently of one another) from the group comprising lower alkyl, such as especially methyl, ethyl or n-propyl, hydroxy-lower alkyl, typically 2-hydroxyethyl, C 3 -C 8 cycloalkyl, especially cyclohexyl, amino-lower afkyl, typically 3-aminopropyl or 4-aminobutyl, N-mono- or N,N-di(lower alkyl)-amino-lower alkyl, typically 3-(N,N-dimethylamino)propyl, amino, N-mono- or N,N-di-lower alkylamino and N-mono- or N,N-di
  • Preferred here is pyrrolidino (1-pyrrolidinyl), piperidino (1-piperidinyl), piperazino (1-piperazinyl), 4-lower al- kylpiperazino, typically 4-methylpiperazino, imidazolino (1-imidazolyl), morpholino (4-morpho- linyl), or also thiomorpholino, S-oxo-thiomorpholino, or S,S-dioxothiomorpholino.
  • An acyl derived from an organic sulfonic acid which is designated Ac 2
  • Ac 2 is especially one with the subformula R°-SO 2 -, wherein R 0 is a hydrocarbyl as defined above in the general and specific meanings, the latter also being generally preferred here.
  • R 0 is a hydrocarbyl as defined above in the general and specific meanings, the latter also being generally preferred here.
  • Especially preferred is lower alkylphenylsulfonyl, especially 4-toluenesulfonyl.
  • Preferred compounds according to the invention are, for example, those wherein R 0 has the following preferred meanings: lower alkyl, especially methyl or ethyl, amino-lower alkyl, wherein the amino group is unprotected or is protected by a conventional amino protecting group - especially by lower alkoxycarbonyl, typically tert-lower alkoxy- carbonyl, for example tert-butoxycarbonyl - e.g.
  • Preferred acyl radicals Ac 1 are acyl radicals of a carboxylic acid which are characterised by the subformula R°-CO-, wherein R 0 has one of the above general and preferred meanings of the hydrocarbyl radical R 0 .
  • Especially preferred radicals R 0 here are lower alkyl, especially methyl or ethyl, amino-lower alkyl, wherein the amino group is unprotected or protected by a conventional amino protecting group, especially by lower al- koxycarbonyl, typically tert-lower alkoxycarbonyl, for example tert-butoxycarbonyl, e.g.
  • a further preferred Acyl Ac 1 is derived from monoesters of carbonic acid and is characterised by the subformula R°-O-CO-.
  • the lower alkyl radicals, especially tert-butyl, are especially preferred hydrocarbyl radicals R 0 in these derivatives.
  • Z is especially lower alkyl, most especially methyl or hydrogen.
  • the compounds of the invention may also be present in the form of pharmaceutically, i.e. physiologically, acceptable salts, provided they contain salt- forming groups.
  • pharmaceutically unacceptable salts may also be used.
  • pharmaceutically acceptable salts are used, and these salts are preferred.
  • compounds of formula I having free acid groups may exist as a salt, preferably as a physiologically acceptable salt with a salt-forming basic component.
  • a salt-forming basic component may be primarily metal or ammonium salts, such as alkali metal or alkaline earth metal salts, for example sodium, potassium, magnesium or calcium salts, or ammonium salts with ammonia or suitable organic amines, especially tertiary monoamines and heterocyclic bases, for example triethylamine, tri-(2-hydroxyethyl)-amine, N-ethylpiperidine or N.N'-dimethylpiperazine.
  • Compounds of the invention having a basic character may also exist as addition salts, especially as acid addition salts with inorganic and organic acids, but also as quaternary salts.
  • compounds which have a basic group, such as an amino group, as a substituent may form acid addition salts with common acids.
  • Suitable acids are, for example, hydrohalic acids, e.g.
  • hydrochloric and hydrobromic acid sulfuric acid, phosphoric acid, nitric acid or perchloric acid, or aliphatic, alicyclic, aromatic or heterocyclic carboxylic or sulfonic acids, such as formic, acetic, propionic, succinic, glycolic, lactic, malic, tartaric, citric, fumaric, maleic, hydroxymaleic, oxalic, pyruvic, phenylacetic, benzoic, p-aminobenzoic, anthranilic, p-hydroxybenzoic, salicylic, p-aminosalicylic acid, pamoic acid, methanesulfonic, ethanesulfonic, hydroxyethanesulfonic, ethylenedisulfonic, halobenzenesulfonic, toluenesulfonic, naphthalenesulfonic acids or sulfanilic acid, and also methionine, tryptophan,
  • any reference hereinbefore and hereinafter to the free compounds is to be understood as referring also to the corresponding salts, and the solvates thereof, for example hydrates, as appropriate and expedient.
  • R- I and R 2 independently of each other are lower alkyl, lower alkyl substituted by halogen, C 6 -C 14 aryl, hydroxy, lower alkoxy, phenyl-lower alkoxy, phenyloxy, lower alkanoyloxy, benzoyloxy, amino, lower alkylamino, lower alkanoylamino, phenyl-lower alkylamino, N,N-di-lower alkylamino, N,N-di-(phenyl-lower alkyl)amino, cyano, mercapto, lower alkylthio, carboxy, lower alkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, N,N-di- lower alkylcarbamoyl, sulfo, lower alkanesulfonyl, lower alkoxysulfonyl, aminosulfonyl, N- lower alkylaminosul
  • Re, R I Q are independently of each other hydrogen, lower alkyl, lower alkenyl or lower alkadienyl, which are each unsubstituted or monosubstituted or polysubsituted, preferably monosubstituted or disubstituted by a substituent independently selected from lower alkyl; hydroxy; lower alkoxy, which may be unsubstituted or mono-, di-, or trisubstituted by (i) heterocyclyl with 4 to 12 ring atoms, which may be unsaturated, wholly saturated, or partly saturated, is monocyclic or bicyclic and may contain up to three heteroatoms selected from nitrogen, oxygen and sulfur, and is most especially pyrrolyl, for example 2-pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, or in a broader sense also thienyl, for example 2- or 3-thienyl, or furyl, for example
  • R 4 is absent for compounds of formula II, hydrogen or CH 3 for compounds of formula I, and
  • R 5 is hydrogen or lower alkyl, especially hydrogen, X stands for 2 hydrogen atoms, for O, or for 1 hydrogen atom and hydroxy; or for 1 hydrogen atom and lower alkoxy;
  • Z is hydrogen or especially lower alkyl, most especially methyl; and for compounds for formula II, either the two bonds characterised by wavy lines are preferably absent in ring A and replaced by 4 hydrogen atoms, and the two wavy lines in ring B each, together with the respective parallel bond, signify a double bond; or also the two bonds characterised by wavy lines are absent in ring B and replaced by a total of 4 hydrogen atoms, and the two wavy lines in ring A each, together with the respective parallel bond, signify a double bond; or both in ring A and in ring B all of the 4 wavy bonds are absent and are replaced by a total of 8 hydrogen atoms; or a salt thereof, if at least one salt-forming group is present.
  • R 3 and R 4 are independently of each other: hydrogen, lower alkyl unsubstituted or mono- or disubstituted, especially monosubstituted, by radicals selected independently of one another from carboxy; lower alkoxycarbonyl; and cyano; or
  • R 4 is hydrogen or -CH 3 , and R 3 is as defined above or preferably R 3 is, acyl of the subformula R°-CO, wherein R 0 is lower alkyl; amino-lower alkyl, wherein the amino group is present in unprotected form or is protected by lower alkoxycarbonyl; tetrahydropyranyloxy-lower alkyl; phenyl; imidazolyl-lower alkoxyphenyl; carboxyphenyl; lower alkoxycarbonylphenyl; halogen-lower alkylphenyl; imidazol-1-ylphenyl; pyrrolidino-lower alkylphenyl; piperazino-lower alkylphenyl; (4-lower alkylpiperazinomethyl)phenyl; morpholino-lower alkylphenyl; piperazinocarbonylphenyl; or (4-lower alkylpiperazino)phenyl; or is
  • R 3 and R 4 are independently of each other hydrogen, lower alkyl unsubstituted or mono- or disubstituted, especially monosubstituted, by radicals selected independently of one another from carboxy; lower alkoxycarbonyl; and cyano; whereby R 4 may also be absent; or
  • R 4 is absent
  • R 3 is acyl from the subformula R°-CO, wherein R 0 is lower alkyl, especially methyl or ethyl; amino-lower alkyl, wherein the amino group is unprotected or protected by lower alkoxycarbonyl, typically tert-lower alkoxycarbonyl, for example tert-butoxycarbonyl, e.g.
  • R 5 is hydrogen or lower alkyl, especially hydrogen, X stands for 2 hydrogen atoms or for O; Z is methyl or hydrogen; and either the two bonds characterised by wavy lines are preferably absent in ring A and replaced by 4 hydrogen atoms, and the two wavy lines in ring B each, together with the respective parallel bond, signify a double bond; or also the two bonds characterised by wavy lines are absent in ring B and replaced by a total of 4 hydrogen atoms, and the two wavy lines in ring A each, together with the respective parallel bond, signify a double bond; or both in ring A and in ring B all of the 4 wavy bonds are absent and are replaced by a total of 8 hydrogen atoms; or a salt thereof, if at least one salt-forming group is present.
  • X 1 hydrogen and 1 hydroxy atom
  • R 1 , R 2 , Rs H
  • R 3 CH 3
  • Z CH 3
  • R 4 is selected from -(CH 2 ) 2 OH; -CH 2 CH(OH)CH 2 OH; -CO(CH 2 ) 2 CO 2 Na; -(CH 2 ) 3 CO 2 H; - COCH 2 N(CHa) 2 ;
  • R 4 is selected from N-[0-(tetrahydropyran-4-yl )-D-lactoyl]; N-[2-(tetrahydro-pyran-4-yloxy)-acetyl)]
  • X 1 hydrogen and 1 hydroxy atom ;
  • R 1 , R 2 , R 5 H;
  • R 3 CH 3 ;
  • Z CH 3 ;
  • R 4 is selected from N-[0-(tetrahydropyran-4-yl )-D-lactoyl]; N-[2-(tetrahydro-pyran-4-yloxy)-acetyl)]
  • CAS means the CHEMICAL ABSTRACTS registry number.
  • MIDOSTAURIN is a derivative of the naturally occurring alkaloid staurosporine, and has been specifically described in the European patent No. 0 296 110 published on December 21 , 1988, as well as in US patent No. 5;093,330 published on March
  • MIDOSTAURIN possesses therapeutic properties, which render it particularly useful as a modulator of transformed cells containing at least one SPTBN 1-FLT3 translocation, and especially in the treatment and prophylaxis of myeloid leukemias, especially atypical chronic myeloid leukemia.
  • This compound shows an unexpected high potency toward treating cancer patients harboring a
  • SPTBN 1-FLT3 translocation which appears to be an activating mutation, as shown in Ba/F3 cell lines harbouring the SPTBN 1-FLT3 translocation construct.
  • STAUROSPORINE DERIVATIVES e.g. MIDOSTAURIN were originally identified as inhibitor of protein kinase C (PKC) (Meyer T, Regenass U, Fabbro D, et al: lnt J
  • FLT3 translocation-mediated signalling pathways and especially in the treatment and prophylaxis of diseases mediated by SPTBN1-FLT3 translocation-associated signalling pathways, especially atypical chronic myeloid leukemia.
  • This compound shows an unexpected high potency toward cell lines harboring SPTBN1-FLT3 translocation, an activating mutation especially in atypical chronic myeloid leukemia.
  • DERIVATIVES for the preparation of a drug for inhibiting SPTBN 1-FLT3 translocation protein product and downstream effects (e.g., mediated by FGFR3, ras, c-fos and/or TGF- beta).
  • the present invention more particularly concerns the use of
  • STAUROSPORINE DERIVATIVES for the preparation of a drug for the treatment of diseases involving SPTBN 1 or Flt3 signalling pathways or SPTBN 1-FLT3 translocation- modulated pathways or SPTBN1-FLT3 translocation gene product.
  • Preferred diseases are myeloid leukemias, including especially atypical chronic myeloid leukemias.
  • the present invention more particularly concerns the use of
  • STAUROSPORINE DERIVATIVES for the preparation of a drug for the treatment of myeloid leukemias, including especially atypical chronic myeloid leukemias.
  • the instant invention provides a method for treating diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product comprising administering to a mammal in need of such treatment a therapeutically effective amount of one or more STAUROSPORINE
  • the instant invention provides a method for treating mammals especially humans suffering from diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product comprising administering to a mammal in need of such treatment a FLT3 receptor tyrosine kinase activity inhibiting amount of ⁇ /-[(9S, 10R, 11 R, 13R)-2,3, 10,11 ,12,13-hexahydro-10-methoxy-9- methyl-1-oxo-9J3-epoxy-1H f 9H-diindolo[1 ,2 > 3-gh:3 ⁇ 2 ⁇ 1 I -lm]pyrrolo[3,4-j][1 ,7]benzodiazonin-
  • the instant invention also concerns a method wherein the therapeutically effective amount of the compound of formula VII is administered to a mammal subject 7 to 4 times a week or about 100 % to about 50% of the days in the time period, for a period of from one to six weeks, followed by a period of one to three weeks, wherein the agent is not administered and this cycle being repeated for from 1 to several cycles.
  • this method is used for treating myeloid leukemia, including especially atypical chronic myeloid leukemia.
  • the instant invention relates to the use of
  • STAUROSPORINE DERIVATIVES for the preparation of a pharmaceutical composition for use in treating diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product.
  • STAUROSPORINE DERIVATIVES have useful pharmacological properties.
  • the activity of the STAUROSPORINE DERIVATIVES especially compounds of formula I or II can be demonstrated, for example, in a single oral administration per day to animals at doses in the range of 5 to 300 or 100 to 200 mg/kg of body weight per day.
  • the STAUROSPORINE DERIVATIVES are therefore very highly suitable for the treatment of diseases, which respond to inhibition of the FLT3 or TGF-beta signalling pathways and / or SPTBN1-FLT3 translocation gene product transcription, e.g., atypical chronic myeloid leukemia.
  • treatment includes both prophylactic or preventative treatment as well as curative or disease suppressive treatment, including treatment of patients at risk of contracting the disease or suspected to have contracted the disease as well as ill patients. This term further includes the treatment for the delay of progression of the disease.
  • curative means efficacy in treating ongoing episodes involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or
  • prophylactic means the prevention of the onset or recurrence of diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or
  • SPTBN 1-FLT3 translocation gene product including the polypeptide encoded by a mutated gene comprising a SPTBN 1-FLT3 translocation.
  • delay of progression means administration of the active compound to patients being in a pre-stage or in an early phase of the disease to be treated, in which patients for example a pre-form of the corresponding disease is diagnosed or which patients are in a condition, e.g. during a medical treatment or a condition resulting from an accident, under which it is likely that a corresponding disease will develop.
  • STAUROSPORINE DERIVATIVES The precise dosage of STAUROSPORINE DERIVATIVES to be employed for inhibiting SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1- FLT3 translocation gene product transcription depends upon several factors including the host, the nature and the severity of the condition being treated, the mode of administration.
  • STAUROSPORINE DERIVATIVES is administered (check preferred administration) parenterally, e.g., intraperitoneally, intravenously, intramuscularly, subcutaneously, intratumorally, or rectally, or enterally, e.g., orally, preferably intravenously or orally, intravenously at a daily dosage of 1-300 mg/kg body weight or, for most larger primates, a daily dosage of 50-5000, preferably 500-3000 mg, in human trials a total dose of 225 mg/day was most presumably the Maximum Tolerated Dose (MTD).
  • MTD Maximum Tolerated Dose
  • a preferred intravenous daily dosage is 1-75 mg/kg body weight or, for most larger primates, a daily dosage of 50-1500 mg.
  • a typical intravenous dosage is 20 mg/kg, three to five times a week.
  • MlDOST AURlN are administered orally, by dosage forms such as microemulsions, soft gels or solid dispersions in dosages up to 150 mg/day, administered in one, two or three times.
  • dosage forms such as microemulsions, soft gels or solid dispersions in dosages up to 150 mg/day, administered in one, two or three times.
  • a small dose is administered initially and the dosage is gradually increased until the optimal dosage for the host under treatment is determined.
  • the upper limit of dosage is that imposed by side effects and can be determined by trial for the host being treated.
  • the STAUROSPORINE DERIVATIVES may be combined with one or more pharmaceutically acceptable carriers and, optionally, one or more other conventional pharmaceutical adjuvants and administered enterally, e.g. orally, in the form of tablets, capsules, caplets, etc. or parenterally, e.g., intraperitoneally or intravenously, in the form of sterile injectable solutions or suspensions.
  • enteral and parenteral compositions may be prepared by conventional means.
  • the infusion solutions according to the present invention are preferably sterile. This may be readily accomplished, e.g. by filtration through sterile filtration membranes.
  • the STAUROSPORINE DERIVATIVES may be formulated into enteral and parenteral pharmaceutical compositions containing an amount of the active substance that is effective for inhibiting SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product, such compositions in unit dosage form and such compositions comprising a pharmaceutically acceptable carrier.
  • the STAUROSPORINE DERIVATIVES can be used alone or combined with at least one other pharmaceutically active compound for use in these pathologies.
  • These active compounds can be combined in the same pharmaceutical preparation or in the form of combined preparations "kit of parts" in the sense that the combination partners can be dosed independently or by use of different fixed combinations with distinguished amounts of the combination partners, i.e., simultaneously or at different time points.
  • the parts of the kit of parts can then, e.g., be administered simultaneously or chronologically staggered, that is at different time points and with equal or different time intervals for any part of the kit of parts.
  • Non-limiting examples of compounds which can be cited for use in combination with STAUROSPORINE DERIVATIVES are cytotoxic chemotherapy drugs, such as cytosine arabinoside, daunorubicin, doxorubicin, cyclophosphamide, VP-16, etc. Further, STAUROSPORINE DERIVATIVES could be combined with other inhibitors of signal transduction or other oncogene-targeted drugs with the expectation that significant synergy would result.
  • compositions comprise a solution or dispersion of compounds of formula I such as MIDOSTAURIN in a saturated polyalkylene glycol glyceride, in which the glycol glyceride is a mixture of glyceryl and polyethylene glycol esters of one or more C8-C18 saturated fatty acids.
  • Composition A [00142] Gelucire 44/14 (82 parts) is melted by heating to 60 DEG C. Powdered
  • MIDOSTAURIN (18 parts) is added to the molten material.
  • the resulting mixture is homogenised and the dispersion obtained is introduced into hard gelatin capsules of different size, so that some contain a 25mg dosage and others a 75mg dosage of the MIDOSTAURIN.
  • the resulting capsules are suitable for oral administration.
  • Composition B is a composition of Composition B:
  • MIDOSTAURIN 14 parts is added to the molten material.
  • the mixture is homogenised and the dispersion obtained is introduced into hard gelatin capsules of different size, so that some contain a 25mg dosage and others a 75mg dosage of the MIDOSTAURIN.
  • the resulting capsules are suitable for oral administration.
  • Gelucire 44/14 available commercially from Gattefosse; is a mixture of esters of C8-C18 saturated fatty acids with glycerol and a polyethylene glycol having a molecular weight of about 1500, the specifications for the composition of the fatty acid component being, by weight, 4-10% capryllc acid, 3-9% capric acid, 40-50% lauric acid, 14-24% myristic acid, 4-14% palmitic acid and 5-15% stearic acid.
  • Gelucire formulation consists of:
  • MIDOSTAURIN 3.0gfilled into a 60 mL Twist off flask
  • a preferred example of soft gel will contain the following Microemulsion:
  • this invention relates to use or method as described herein, wherein the daily effective amount of the compound of formula VII, is 100 to 300 mg, preferably 125 mg to 250 mg most preferably 220 to 230 mg, preferably 225 mg.
  • the compound of formula VII is administered once, two or three times a day, for a total dose of 100 to 300 mg daily.
  • the compound of formula VII is administered three times a day, for a total dose of 220 to 230 preferably 225 mg daily, and preferably at a dose per administration of 70 to 80 mg, preferably 75 mg.
  • this invention relates to an article of manufacture comprising packaging material, and ⁇ /-[(9S,10/?,11R13R)-2,3,10,11 ,12,13-hexahydro-10- methoxy-9-methyl-1 -oxo-9, 13-epoxy-1 W,9/-/-diindolo[1 ⁇ .S-gtrS 1 ⁇ 1 , 1 '-lm]pyrrolo[3,4- j][1,7]benzodiazonin-11-yl]-/V-methylbenzamide of the formula (VII) or a pharmaceutically acceptable salts thereof, contained within said packaging material, wherein said packaging material comprises label directions which indicate that said compound of formula (VII), or said pharmaceutically-acceptable salt, is to be administered to mammals suffering from diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product, in an amount from 50 to 500 mg, preferably
  • an article of manufacture wherein the compound of formula VII is administered three times a day, for a total dose of 220 to 230mg, preferably 225 mg daily, and preferably a dose of 70 to 80 mg most preferably 75 mg per administration for treating myeloid leukemia, including especially atypical chronic myeloid leukemia.
  • a preferred embodiment relates to an article of manufacture comprising softgel capsules containing 25 mg of the compound of formula VII.
  • the application of involving SPTBN 1-FLT3 translocation-associated genotypes and/or haplotypes that correlate with efficacious drug responses will be used to select patients for therapy of existing diseases.
  • the application of genotypes and/or haplotypes that correlate with a predisposition for disease will likewise be used to select patients for preventative therapy.
  • a further aspect of the invention is a method for determining whether a mammal, especially a human, has a disease involving SPTBN1-FLT3 translocation, and preferably whether atypical chronic myeloid leukemia cells in such a mammal have a SPTBN 1-FLT3 translocation.
  • Methods for detecting such SPTBN 1-FLT3 translocation include, but are not limited, to the following molecular genetic techniques: reverse- transcriptase (RT) and genomic polymerase chain reaction (PCR) using primer sequences derived from the SPTBN1-FLT3 translocation sequence encoding the SPTBN1-FLT3 fusion polypeptide (see FIG. 1 , panels 1A-1D).
  • the forward primer sequence is preferably derived from exons 1-3 of SPTBN1 and reverse primer is preferably derived from sequences downstream from exon 12 in FLT3.
  • the translocation genomic breakpoint can be amplified by PCR with forward and reverse primers derived from exon 3 of SPTBN 1 and FLT3 exon 13.
  • SPTBN1 and FLT3 rearrangement may also be identified through fluorescent in situ hybridisation (FISH), using fosmid or bacterial artificial chromosome clones (BAC) around the breakpoint of the FLT3 and SPTBN 1 genes. Suitable clones and labelling techniques will be apparent to those skilled in the art.
  • the SPTBN 1-FLT3 translocation may be detected at the protein level, for example by determining that the molecular size of the FLT3 is modified in the affected cells or that FLT3 protein may be isolated using antibodies, or other specific binding entities, specific to SPTBN1.
  • a further aspect of the invention is a polynucleotide encoding a gene product comprising a fusion of the SPTBN1 and FLT3 genes, wherein the polynucleotide comprises genomic sequence of the fused SPTBN1-FLT3 translocation, and the gene product thereof, wherein the gene product is the RNA or its respective cDNA comprising the transcribed fusion gene sequence or the translated polypeptide encoded by the transcribed fusion gene sequence, and their usage as reagents in diagnostic assays for SPTBN1-FLT3 translocations and screening assays for agents capable of inhibiting the function of SPTBN 1- FLT3 gene fusion products or determining the effect of such gene products.
  • an isolated nucleic acid molecule of the invention is at least 6 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the SPTBN1-FLT3 gene fusion nucleotide sequence.
  • the nucleic acid is at least 10, 25, 50, 100, 250, 500, 1000, 2000 or 2165 nucleotides in length.
  • an isolated nucleic acid molecule of the invention hybridizes to the coding region of the SPTBN1-FLT3 fusion gene.
  • an isolated nucleic acid molecule of the invention hybridizes to the messenger RNA or corresponding cDNA.
  • hybridizes under stringent conditions refers to conditions for hybridization and washing under which nucleotide sequences at least 70% homologous to each other typically remain hybridized to each other.
  • stringent hybridization conditions refers to conditions under which a probe, primer or oligonucleotide will hybridize to its target sequence, but to no other sequences. Stringent conditions are sequence-dependent and selection of appropriate hybridization conditions are within the skill of one skilled in the art. Longer sequences hybridize specifically at higher temperatures than shorter sequences. Generally, stringent conditions are selected to be about 5 0 C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH.
  • Tm thermal melting point
  • the Tm is the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50% of the probes complementary to the target sequence hybridize to the target sequence at equilibrium.
  • Stringent conditions are known to those skilled in the art and can be found in Ausubel et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, N.Y., 6.3.1-6.3.6 (1989).
  • the conditions are such that sequences at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other.
  • Typical stringent conditions include, but are not limited to, e.g., a salt concentration of less than about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0 to 8.3 and wherein the temperature is at least about 3O 0 C for short probes, primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about 60 0 C for longer probes, primers and oligonucleotides.
  • Stringent conditions may also be achieved with the addition of destabilizing agents, such as formamide, using standard procedures known to one skilled in the art.
  • stringent hybridization conditions is hybridization in a high salt buffer comprising 6xSSC, 50 mM Tris-HCI (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/ml denatured salmon sperm DNA at 65 0 C, followed by one or more washes in 0.2xSSC, 0.01% BSA at 5O 0 C.
  • Other conditions of high stringency, moderate stringency and low stringency that may be used are well-known in the art. See, e.g., Ausubel et al.
  • Naturally-occurring allelic variants of the SPTBN1 and FLT3 gene sequence may exist in the population, and may translocate to generate the SPTBN1-FLT3 fusion gene. Such changes can be introduced into the SPTBN1-FLT3 fusion gene nucleotide sequence of SEQ ID NO: 1 , on occasion leading to changes in the amino acid sequence of the encoded SPTBN1-FLT3 fusion protein, without altering the functional ability of the SPTBN1-FLT3 fusion protein. Nucleotide substitutions leading to amino acid substitutions at "non-essential" amino acid residues can be made in SEQ ID NO:1 and inserted into a plasmid or vector known in the art.
  • the isolated nucleic acid molecule comprises a nucleotide sequence encoding a SPTBN1-FLT3 fusion protein, wherein the protein comprises an amino acid sequence at least about 85% identical, 90% identical, 95% identical, up to 99 % identical to the SPTBN 1-FLT3 fusion polypeptide sequence of SEQ ID NO:2.
  • the protein encoded by the nucleic acid molecule is at least about 85% identical to the SPTBN 1-FLT3 fusion polypeptide, more preferably at least about 90% homologous, and most preferably at least about 95% homologous to the given SPTBN 1-FLT3 fusion polypeptide.
  • modified SPTBN1-FLT3 fusion sequences can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of the SPTBN 1-FLT3 gene fusion, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein.
  • Mutations can be introduced into SEQ ID NO:1 by standard techniques, e.g., site-directed mutagenesis and PCR-mediated mutagenesis.
  • conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues.
  • a "conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art.
  • amino acids with basic side chains e.g., K, R or H, using the one amino acid code
  • acidic side chains e.g., D, E
  • uncharged polar side chains e.g., F, N, Q, S, T, Y, C
  • nonpolar side chains e.g., A, V, L, I, P, F, M, W
  • beta-branched side chains e.g., T, V, I
  • aromatic side chains e.g., Y, F, W, H.
  • a predicted nonessential amino acid residue in SPTBN1-FLT3 fusion polypeptide is replaced with another amino acid residue from the same side chain family.
  • additional contemplated conserved amino acid substitutions include, e.g., replacement of one amino acid in a group with any other amino acid listed in the same group, wherein exemplary conserved groups include but are not limited to: FYW, HY, MILV, MILF, NDEQ, NEQK 1 NHQK 1 QHRK and STA.
  • mutations can be introduced randomly along all or part of a SPTBN1-FLT3 gene fusion coding sequence, e.g., by saturation mutagenesis, and the resultant mutants can be screened for biological activity to identify mutants that retain the activity of the SPTBN1-FLT3 fusion polypeptide of SEQ ID NO:2.
  • the encoded modified SPTBN 1-FLT3 fusion polypeptide can be expressed by any recombinant technology known in the art and the activity of the protein can be determined.
  • the modified SPTBN1-FLT3 fusion polypeptide can be assayed for the ability of the modified SPTBN 1-FLT3 fusion polypeptide to transform cell lines, such as the Ba/F3 cells in the Examples.
  • the DNA sequence homology referred to above may be determined as the degree of identity between two sequences indicating a derivation of the first sequence from the second.
  • the homology may suitably be determined by means of computer programs known in the art, such as GAP provided in the GCG program package. See, e.g., Needleman and Wunsch, J MoI Biol 1970 48: 443-453. Homology may also be determined by computer analysis such as, without limitations, the ClustalX computer alignment program. See, e.g., Thompson et al. Nucleic Acids Res. 1997, 25 (24): 4876-82.
  • sequence identity refers to the degree to which two polynucleotide sequences are identical on a nucleotide-by-nucleotide basis over a particular region of comparison.
  • percentage of sequence identity is calculated by comparing two optimally aligned sequences over that region of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the region of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity.
  • substantially identical denotes a characteristic of a polynucleotide sequence, wherein the polynucleotide comprises a sequence that has at least 80 percent sequence identity, preferably at least 85 percent identity and often 90 to 95 percent sequence identity, more usually at least 99 percent sequence identity as compared to a reference sequence over a comparison region.
  • SPTBN 1-FLT3 polypeptides or polypeptide fragments of the invention are used to produce SPTBN1-FLT3 fusion-specific antibodies.
  • a " SPTBN1- FLT3 fusion-specific antibody is an antibody, e.g., a polyclonal antibody or a monoclonal antibody, that is immunoreactive to a SPTBN 1-FLT3 fusion polypeptide or polypeptide fragment, or that binds with specificity to an epitopes of a SPTBN1-FLT3 fusion polypeptides.
  • polyclonal and monoclonal antibodies are well known in the art.
  • Polyclonal antibodies may in particular be obtained as described by, e.g., Green et al.,: “Production of Polyclonal Antisera” in IMMUNOCHEMICAL PROTOCOLS (Manson, Ed.); Humana Press, 1992, pages 1-5; by Coligan et al. in CURRENT PROTOCOLS IN IMMUNOLOGY, 1992, Section 2.4.1, and by Harlow and Lane (Eds.) in "ANTIBODIES; A LABORATORY MANUAL” Cold Spring Harbor Lab. Press 1988. These protocols are hereby incorporated by reference.
  • Monoclonal antibodies may in particular be obtained as described by, e.g., Kohler & Milstein, Nature. 1975, 256:495; Coligan et al., in CURRENT PROTOCOLS IN IMMUNOLOGY, 1992, Sections 2.5.1-2.6.7; and Harlow et al., in ANTIBODIES: A LABORATORY MANUAL; Cold Spring Harbor, Pub., 1988, page 726; which protocols are hereby incorporated by reference.
  • Antibodies that bind to the SPTBN 1-FLT3 fusion polypeptide can be prepared using an intact polypeptide or fragments containing small peptides of interest as the immunizing antigen.
  • the polypeptide used to immunize an animal may be obtained by recombinant DNA techniques or by chemical synthesis, and may optionally be conjugated to a carrier protein.
  • Commonly used carrier proteins which are chemically coupled to the peptide include keyhole limpet hemocyanin (KLH), thyroglobulin, bovine serum albumin (BSA), and tetanus toxoid.
  • KLH keyhole limpet hemocyanin
  • BSA bovine serum albumin
  • the coupled peptide may then be used to immunize the animal, which may in particular be a mouse, a rat, a hamster or a rabbit.
  • EXAMPLE 1 EXAMPLE 1 :
  • Sequences used to amplify the fusion by RT-PCR as a single step include forward primer SPTBN1 1F (5'-aga aga aga cct tea cca agt-3')(SEQ ID NO:3) and FLT3 1R (5'-tga ace agg cat get gga at-3')(SEQ ID NO:4).
  • translocation genomic breakpoint was amplified by PCR with forward and reverse primers derived from exon 3 of SPTBN1 (5'-cca tga cat cgt gga tgg aaa cc-3')(SEQ ID NO:5) and FLT3 exon 13 (5'-tgt ace ate tgt age tgg ctt tc-3')(SEQ ID NO:6).
  • the patient was initially treated with hydroxyurea and subsequently underwent an unrelated donor bone marrow transplant. She relapsed cytogenetically at 4 years but responded to donor lymphocyte infusion (DLI), achieving sustained cytogenetic and molecular (nested RT-PCR) remission.
  • DLI donor lymphocyte infusion
  • the SPTBN1-FLT3 fusion gene is predicted to be translated into a 570 amino acid chimeric protein that retains two coiled-coil domains from SPTBN 1 and 424 amino acids from FLT3, including the entire tyrosine kinase domain. Since the t(2; ' 13) is readily visible by cytogenetic analysis but has not been reported previously it seems likely that SPTBN1-FLT3 is uncommon.

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Abstract

The present invention relates to the use of STAUROSPORINES DERIVATIVES for the preparation of a drug for the treatment of diseases involving SPTBN1-FLT3 translocation- modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product, especially for the curative and/or prophylactic treatment of myeloid leukemia, including especially atypical chronic myeloid leukemia, and to a method of treating diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product.

Description

TRANSLOCATION AND MODULATORS
FIELD OF THE INVENTION
[001] This invention relates generally to the analytical testing of tissue samples in vitro, and more particularly to aspects of genetic mutations and translocations of SPTBN 1 with FLT3.
BACKGROUND OF THE INVENTION
[002] Conventional medical approaches to diagnosis and treatment of disease is based on clinical data alone, or made in conjunction with a diagnostic test. Such traditional practices often lead to therapeutic choices that are not optimal for the efficacy of the prescribed drug therapy or to minimize the likelihood of side effects for an individual subject. Therapy specific diagnostics (a.k.a., theranostics) is an emerging medical technology field, which provides tests useful to diagnose a disease, choose the correct treatment regime and monitor a subject's response. That is, theranostics are useful to predict and assess drug response in individual subjects, i.e., individualized medicine. Theranostic tests are also useful to select subjects for treatments that are particularly likely to benefit from the treatment or to provide an early and objective indication of treatment efficacy in individual subjects, so that the treatment can be altered with a minimum of delay. Theranostics are useful in clinical diagnosis and management of a variety of diseases and disorders, which include, but are not limited to, e.g., cardiovascular disease, cancer, infectious diseases, Alzheimer's disease and the prediction of drug toxicity or drug resistance. Theranostic tests may be developed in any suitable diagnostic testing format, which include, but is not limited to, e.g., immunoassay, immunohistochemical tests, clinical chemistry, cell-based technologies, and nucleic acid tests.
[003] Progress in pharmacogenomics and pharmacogenetics, which establishes correlations between responses to specific drugs and the genetic profile of individual patients and/or their tumours, is the foundation to the development of new theranostic approaches. As such, there is a need in the art for the evaluation of patient-to-patient variations and tumour mutations in gene sequence and gene expression. A common form of genetic profiling relies on the identification of DNA sequence variations that lead to patient- to-patient variation in individual drug response. It is well established in the art that acquired DNA changes (mutations) are responsible, alone or in part, for pathological processes. It follows that, there is a need art to identify and characterize genetic mutations that are useful in identifying the genotypes of subjects and their tumours associated with drug responsiveness, side effects, or optimal dose.
[004] Spectrin is composed of alpha- and beta- subunits that form antiparallel dimers that self associate, giving rise to the cytoskeletal spectrin tetramer. Beta subunits, such as spectrin beta nonerythrocytic 1 ("SPTBN1" OMIM accno. 182790), contain most of the spectrin binding activity (Hayes et al., J. Cell ScL 113: 2023-2034 (2000). Disruption of SPTBN1 leads to the disruption of TGF-beta (OMIM accno. 190180) signaling by Smad proteins in mice. SPTBN 1 null (-/-) mice exhibit a phenotype similar to Smad2/Smad3 double heterozygous mice, with midgestational death due to gastrointestinal, liver, neural, and heart defects. TGF-beta triggers phosphorylation and association of SPTBN1 with Smad3 and Smad4, followed by nuclear translocation. Tang et al. Science 299: 574-577 (2003). SPTBN1 deficiency results in mislocalization of Smad3 and Smad4 and loss of TGF-beta- dependent transcriptional response. The SPTBN1 deficient phenotype could be rescued by overexpression of the carboxy-terminal region of SPTBN1 , which suggests an unexpected molecular link between SPTBN1 and the TGF-beta signaling pathway. Tang et al. (2003). [005] FLT3 kinase is a member of the type III receptor tyrosine kinase (RTK) family.
FLT3 (fms-like tyrosine kinase) is also known as FLk-2 (fetal liver kinase 2). Aberrant expression of the FLT3 gene has been documented in both adult and childhood leukemias including acute myeloid leukemia (AML), AML with trilineage myelodysplasia (AML/TMDS), acute lymphoblastic leukemia (ALL), and myelodysplastic syndrome (MDS). [006] Activating mutations of the FLT3 receptor have been found in about 35% of patients with acute myeloblasts leukemia (AML), and are associated with a poor prognosis. The most common mutation involves an in-frame duplication within the juxtamembrane domain, with an additional 5-10% of patients having a point mutation at asparagine 835. Both of these mutations are associated with constitutive activation of the tyrosine kinase activity of FLT3, and result in proliferation and viability signals in the absence of ligand. Patients expressing the mutant form of the receptor have been shown to have a decreased chance for cure. Thus, there is accumulating evidence for a role for hyperactivated (mutated) FLT3 kinase activity in human cancers.
[007] Accordingly, there is a need in the art for additional information about the relationship between SPTBN 1-FLT3 gene translocations and cancer. SUMMARY OF THE INVENTION
[008] The invention provides for the use of a FLT3 modulating agent in the manufacture of a medicament for the treatment of cancer in a selected patient population. The patient population is selected on the basis of the genotype of the patients SPTBN 1- FLT3 genetic translocation indicative of efficacy of the FLT3-modulating agent in treating cancer. In one embodiment, the cancer can be atypical chronic myeloid leukemia. [009] The invention further provides a method for treating cancer in a subject. The genotype or haplotype of a subject is obtained to identify patients with a translocation between the SPTBN1-FLT3 gene loci, so that the genotype and/or haplotype is indicative of a propensity of the cancer to respond to the drug. Then, the anti-cancer therapy is administered to the subject.
BRIEF DECRIPTION OF DRAWINGS
[0010] FIG. 1 (panels 1A-1 D) provides the full length cDNA (SEQ ID NO:1) sequence of the SPTBN1-FLT3 gene fusion and its encoded 570 aa polypeptide (SEQ ID NO:2).
DETAILED DESCRIPTION OF THE INVENTION
[0011] Leukemia is a progressive, malignant disease of the blood-forming organs, characterized by distorted proliferation and development of leukocytes and their precursors in the blood and bone marrow. Leukemia is classified as acute or chronic, which refer to the degree of cell differentiation, and as myelogenous or lymphocytic, which refer to the predominant type of cell involved. Myeloid leukemia is a variety of leukemia in which some types of white blood cells, originating in the myeloid tissue of the bone marrow, proliferate and suppress healthy red and white blood cells. The present invention relates to the efficacy of staurosporine derivatives in the treatment of atypical chronic myeloid leukemia containing a SPTBN1-FLT3 translocation.
[0012] Definitions. The definitions of certain genetic terms as used in this specification may be found in the glossary provided by the U.S. Department of Energy, Office of Science, Human Genome Project (http://www.ornl.gov/sci/techresources/ Human_Genome/glossary/).
FLT3 Kinase Inhibitors
[0013] The present invention relates to the use of staurosporine derivatives of formula A, B, C, D, I1 II, III, IV, V, Vl and VII: (hereinafter: "STAUROSPORINE DERIVATIVES") for the preparation of a drug for the treatment of diseases involving a SPTBN1-FLT3 translocation and associated signalling pathways, especially for the curative and/or prophylactic treatment of atypical chronic myeloid leukemia, and to a method of treating diseases involving SPTBN1-FLT3 translocation-associated signalling pathways.
[0014] The invention relates to the use of staurosporine derivatives of formula,
Figure imgf000005_0001
Figure imgf000005_0002
wherein Ri , and R2 are, independently of one another, unsubstituted or substituted alky!, hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubsti- tuted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl; n and m are, independently of one another, a number from and including 0 to and including 4;
R5 is hydrogen, an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms in each case, or a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms in each case, and in each case up to 9 heteroatoms, or acyl with up to 30 carbon atoms;
X stands for 2 hydrogen atoms; for 1 hydrogen atom and hydroxy; for O; or for hydrogen and lower alkoxy; Q and Q' are independently a pharmaceutically acceptable organic bone or hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N- mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N- mono- or N,N-di-substituted aminosulfonyl; or a salt thereof, if at least one salt-forming group is present, or hydrogenated derivative thereof, for the preparation of a pharmaceutical composition for the treatment of diseases involving SPTBN1-FLT3 translocation associated signalling pathways.
[0015] The term "organic bone" as used herein refers to a pharmacologically acceptable organic chemical structure, such as but not limited to hydrocarbyl radical or an acyl radical Ac, which radicals preferably have a maximum of 30 carbon atoms. [0016] The hydrocarbyl radical (hydrocarbon radical) is an acyclic (aliphatic), , carbocyclic or carbocyclic-acyclic hydrocarbon radical having a maximum total number of carbon atoms of preferably 30 and, especially, 18, which may be saturated or unsaturated, unsubstituted or substituted. It may also contain instead of one, two or more carbon atoms the same or different hetero atoms, such as, especially, oxygen, sulphur and nitrogen, in the acyclic and/or cyclic moiety; in the latter case it is referred to as a heterocyclic radical (heterocyclyl radical) or a heterocyclic-acyclic radical.
[0017] Unsaturated radicals are those that contain one or more, especially conjugated and/or isolated, multiple bonds (double and/or triple bonds). The term "cyclic radicals" also includes aromatic radicals, for example those in which at least one 6- membered carbocyclic ring or one 5- to 8-membered heterocyclic ring contains the maximum number of non-cumulated double bonds. Carbocyclic radicals in which at least one ring is in the form of a 6-membered aromatic ring (that is to say a benzene ring) are referred to as aryl radicals.
[0018] An acyclic unsubstituted hydrocarbon radical is especially a straight or branched lower alkyl, lower alkenyl, lower alkadienyl or lower alkynyl radical. In corresponding unsaturated radicals, the double bond is located especially in a position higher than the alpha-position to the free valency.
[0019] A carbocyclic hydrocarbon radical is especially a mono-, bi- or polycyclic cycloalkyl, cycloalkenyl or cycloalkadienyl radical, or a corresponding aryl radical. Preferred are radicals having a maximum of 14, especially 12, ring carbon atoms and having 3- to 8- membered, preferably 5- to 7-membered, especially 6-membered, rings; they may also carry one or more, for example two, acyclic radicals, for example those mentioned above, and especially lower alkyl radicals, or other carbocyclic radicals. Carbocyclic-acyclic radicals are those in which an acyclic radical, especially one having a maximum of 10, preferably a maximum of 6, carbon atoms, such as, especially, methyl, ethyl or vinyl, carries one or more of the carbocyclic, optionally aromatic radicals defined above. Mention is made especially of cycloalkyl-lower alkyl and aryl-lower alkyl radicals, and also analogues thereof unsaturated in the ring and/or chain, that carry the ring at the terminal carbon atom of the chain. [0020] Linkers between the acyclic (aliphatic) or carbocyclic radicals may be selected from, but not limited to, straight or branched lower alkyl, lower alkenyl, lower alkadienyl or lower alkynyl radical, etherified or esterified hydroxy, amino, -O-, -S-, carbonyl, -NO-, -SO-, carbonyldioxy, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl. [0021] The term "leukemia" as used herein refers to the clonal proliferation in bone marrow or blood of one or more of the haematopoietic lineages (World Health Organization Classification of Tumours, 2001). Myeloid leukemia is the clonal proliferation of one or more of the myeloid lineages. Chronic myeloid leukemia ("CML") is a myeloproliferative disease that originates in an abnormal bone marrow stem cell and is consistently associated with the Philadelphia (Ph) chromosome and/or the BCR-ABL fusion gene. The disease is bi- or triphasic: an initial indolent chronic phase (CP-CML) is followed by one or both of the aggressive transformed stages, accelerated phase (CML-AP) or blast phase (CML-BP). "Atypical chronic myeloid leukemia" ("aCML") is a clonal leukemic disorder with myeloproliferative features like CML that does not have the Philadelphia chromosome or the BCR-ABL fusion gene.
[0022] Throughout the present specification and claims "leukemia" means preferably atypical chronic myeloid leukemia.
[0023] The invention relates in particular to the use of staurosporines derivatives of formula,
Figure imgf000008_0001
(II) which is the partially hydrogenated derivative of compound (I)1
Figure imgf000008_0002
Figure imgf000008_0003
wherein Ri and R2, are, independently of one another, unsubstituted or substituted alkyl, hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubsti- tuted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl; n and m are, independently of one another, a number from and including 0 to and including 4; n' and m' are, independently of one another, a number from and including 0 to and including 4; R3, R4, Rs and R10 are, independently of one another, hydrogen, -O ", acyl with up to 30 carbon atoms, an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms in each case, a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms in each case, and in each case up to 9 heteroatoms, an acyl with up to 30 carbon atoms, wherein R4 may also be absent; or if R3 is acyl with up to 30 carbon atoms, R4 is not an acyl; p is 0 if R4 is absent, or is 1 if R3 and R4 are both present and in each case are one of the aforementioned radicals; R5 is hydrogen, an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms in each case, or a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms in each case, and in each case up to 9 heteroatoms, or acyl with up to 30 carbon atoms; R7, R6 and R9 are acyl or -(lower alkyl) -acyl, unsubstituted or substituted alkyl, hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy.carbonyl, carbonyldioxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl; X stands for 2 hydrogen atoms; for 1 hydrogen atom and hydroxy; for O; or for hydrogen and lower alkoxy;
Z stands for hydrogen or lower alkyl; and either the two bonds characterised by wavy lines are absent in ring A and replaced by 4 hydrogen atoms, and the two wavy lines in ring B each, together with the respective parallel bond, signify a double bond; or the two bonds characterised by wavy lines are absent in ring B and replaced by a total of
4 hydrogen atoms, and the two wavy lines in ring A each, together with the respective parallel bond, signify a double bond; or both in ring A and in ring B all of the 4 wavy bonds are absent and are replaced by a total of 8 hydrogen atoms; or a salt thereof, if at least one salt-forming group is present for the preparation of a pharmaceutical composition for the treatment of diseases involving SPTBN1-FLT3 translocation associated signalling pathways.
[0024] The general terms and definitions used preferably have hereinbefore and hereinafter the following meanings:
[0025] The prefix "lower" indicates that the associated radical preferably has up to and including a maximum of 7 carbon atoms, especially up to and including a maximum of 4 carbon atoms.
[0026] Lower alkyl is especially methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, and also pentyl, hexyl, or heptyl.
[0027] Unsubstituted or substituted alkyl is preferably Ci-C20alkyl, especially lower alkyl, typically methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, which is unsubstituted or substituted especially by halogen, such as fluorine, chlorine, bromine, or iodine, C6-C14aryl, such as phenyl or naphthyl, hydroxy, etherified hydroxy, such as lower alkoxy, phenyl-lower alkoxy or phenyloxy, esterified hydroxy, such as lower alkanoyloxy or benzoyloxy, amino, mono- or disubstituted amino, such as lower alkylamino, lower alkanoylamino, phenyl-lower alkylamino, N, N-di-lower alkylamino, N,N-di-(phenyl-lower alkyl)amino, cyano, mercapto, substituted mercapto, such as lower alkylthio, carboxy, esterified carboxy, such as lower alkoxycarbonyl, carbamoyl, N-mono- or N,N-disubstituted carbamoyl, such as N-lower alkylcarbamoyl or N, N-di-lower alkylcarbamoyl, sulfo, substituted sulfo, such as lower alkanesulfonyl or lower alkoxysulfonyl, aminosulfonyl or N- mono- or N,N-disubstituted aminosulfonyl, such as N-lower alkylaminosulfonyl or N, N-di- lower alkylaminosulfonyl.
[0028] Halogen is preferably fluorine, chlorine, bromine, or iodine, especially fluorine or chlorine.
[0029] Etherified hydroxy is especially lower alkoxy, C6-C14aryloxy, such as phenyloxy, or C6-C14aryl-lower alkoxy, such as benzyloxy.
[0030] Esterified hydroxy is preferably lower alkanoyloxy or C6-C14arylcarbonyloxy, such as benzoyloxy.
[0031] Mono- or disubstituted amino is especially amino monosubstituted or disubstituted by lower alkyl,
Figure imgf000010_0001
Ce-Ci4aryl-lower alkyl, lower alkanoyl, or C6-Ci 2arylcarbonyl • )
[0032] Substituted mercapto is especially lower alkylthio, C6-C14arylthio, C6-C14aryl- lower alkylthio, lower alkanoylthio, or C6-Ci4aryl-lower alkanoylthio.
[0033] Esterified carboxy is especially lower alkoxycarbonyl, C6-C14aryl-lower alkoxy- carbonyl or
Figure imgf000011_0001
[0034] N-Mono- or N,N-disubstituted carbamoyl is especially carbamoyl N- monosubstituted or N,N-disubstituted by lower alkyl, C6-C14aryl or C6-C14aryl-lower alkyl. [0035] Substituted sulfonyl is especially C6-Ci4arylsulfonyl, such as toluenesulfonyl,
C6-Ci4aryl-lower alkanesulfonyl or lower alkanesulfonyl.
[0036] N-Mono- or N,N-disubstituted aminosulfonyl is especially aminosulfonyl N- monosubstituted or N,N-disubstituted by lower alkyl, C6-C14aryl or C6-Ci4aryl-lower alkyl. [0037] C6-C14Aryl is an aryl radical with 6 to 14 carbon atoms in the ring system, such as phenyl, naphthyl, fluorenyl, or indenyl, which is unsubstituted or is substituted especially by halogen, such as fluorine, chlorine, bromine, or iodine, phenyl or naphthyl, hydroxy, lower alkoxy, phenyl-lower alkoxy, phenyloxy, lower alkanoyloxy, benzoyloxy, amino, lower alkylamino, lower alkanoylamino, phenyl-lower alkylamino, N,N-di-lower alkylamino, N,N-di- (phenyl-lower alkyl)amino, cyano, mercapto, lower alkylthio, carboxy, lower alkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, N,N-di-lower alkylcarbamoyl, sulfo, lower alkanesulfonyl, lower alkoxysulfonyl, aminosulfonyl, N-lower alkylaminosulfonyl, or N,N-di-lower alkylamino- sulfonyl.
[0038] The indices n and m are in each case preferably 1 , 2 or especially 0. In general, compounds of formula I in which n and m are in each case 0 (zero) are especially preferred.
[0039] An aliphatic carbohydrate radical with up to 29 carbon atoms R3, R4,. R8 or Ri0, which is substituted by acyclic substituents and preferably has a maximum of 18, especially a maximum of 12, and as a rule not more than 7 carbon atoms, may be saturated or unsaturated and is especially an unsubstituted or a straight-chain or branched lower alkyl, lower alkenyl, lower alkadienyl, or lower alkinyl radical substituted by acyclic substituents. Lower alkyl is, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, and also n-pentyl, isopentyl, n-hexyl, isohexyl and n-heptyl; lower alkenyl is, for example, allyl, propenyl, isopropenyl, 2- or 3-methallyl and 2- or 3-butenyl; lower alkadienyl is, for example, 1-penta-2,4-dienyl; lower alkinyl is, for example, propargyl or 2-butinyl. In corresponding unsaturated radicals, the double bond is especially located in a position higher than the α-position in relation to the free valency. Substituents are especially the acyl radicals defined hereinbelow as substituents of R0, preferably free or esterified carboxy, such as carboxy or lower alkoxycarbonyl, cyano or di-lower alkylamino. [0040] A carbocyclic or carbocyclic-aliphatic radical R3, R4, R8 or R10 with up to 29 carbon atoms in each case is especially an aromatic, a cycloaliphatic, a cycloaliphatic- aliphatic, or an aromatic-aliphatic radical which is either present in unsubstituted form or substituted by radicals referred to hereinbelow as substituents of R0. An aromatic radical (aryl radical) R3 or R4 is most especially a phenyl, also a naphthyl, such as 1- or 2-naphthyl, a biphenylyl, such as especially 4-biphenylyl, and also an anthryl, fluorenyl and azulenyl, as well as their aromatic analogues with one or more saturated rings, which is either present in unsubstituted form or substituted by radicals referred to hereinbelow as substituents of R0. Preferred aromatic-aliphatic radicals are aryl-lower alkyl- and aryl-lower alkenyl radicals, e.g. phenyl-lower alkyl or phenyl-lower alkenyl with a terminal phenyl radical, such as for example benzyl, phenethyl, 1-, 2-, or 3-phenylpropyl, diphenylmethyl (benzhydryl), trityl, and cinnamyl, and also 1- or 2-naphthylmethyl. Of aryl radicals carrying acyclic radicals, such as lower alkyl, special mention is made of o-, m- and p_-tolyl and xylyl radicals with variously situated methyl radicals.
[0041] A cycloaliphatic radical R3, R4, R8 or R10 with up to 29 carbon atoms is especially a substituted or preferably unsubstituted mono-, bi-, or polycyclic cycloalkyl-, cycloalkenyl-, or cycloalkadienyl radical. Preference is for radicals with a maximum of 14, especially 12, ring-carbon atoms and 3- to 8-, preferably 5- to 7-, and most especially 6- member rings which can also carry one or more, for example two, aliphatic hydrocarbon radicals, for example those named above, especially the lower alkyl radicals, or other cycloaliphatic radicals. Preferred substituents are the acyclic substituents named hereinbelow for R0.
[0042] A cycloaliphatic-aliphatic radical R3, R4, R8 or Ri0 with up to 29 carbon atoms is a radical in which an acyclic radical, especially one with a maximum of 7, preferably a maximum of 4 carbon atoms, such as especially methyl, ethyl, and vinyl, carries one or more cycloaliphatic radicals as defined hereinabove. Special mention is made of cycloalkyl-lower - alkyl radicals, as well as their analogues which are unsaturated in the ring and/or in the chain, but are non-aromatic, and which carry the ring at the terminal carbon atom of the chain. Preferred substituents are the acyclic substituents named herein below for R0. [0043] Heterocyclic radicals R3, R4, R8 or R10 with up to 20 carbon atoms each and up to 9 heteroatoms each are especially monocyclic, but also bi- or polycyclic, aza-, thia-, oxa-, thiaza-, oxaza-, diaza-, triaza-, or tetrazacyclic radicals of an aromatic character, as well as corresponding heterocyclic radicals of this type which are partly or most especially wholly saturated, these radicals - if need be - possibly carrying further acyclic, carbocyclic, or heterocyclic radicals and/or possibly mono-, di-, or polysubstituted by functional groups, preferably those named hereinabove as substituents of aliphatic hydrocarbon radicals. Most especially they are unsubstituted or substituted monocyclic radicals with a nitrogen, oxygen, or sulfur atom, such as 2-aziridinyl, and especially aromatic radicals of this type, such as pyrryl, for example 2-pyrryl or 3-pyrryl, pyridyl, for example 2-, 3-, or 4-pyridyl, and also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl; analogous bicyclic radicals with an oxygen, sulfur, or nitrogen atom are, for example, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, or benzothienyl, typically 2- or 3- benzothienyl; preferred monocyclic and bicyclic radicals with several heteroatoms are, for example, imidazolyl, typically 2- or 4-imidazolyl, pyrimidinyl, typically 2-or 4-pyrimidinyl, oxazolyl, typically 2-oxazolyl, isoxazolyl, typically 3-isoxazolyl, or thiazolyl, typically 2- thiazolyl, and benzimidazolyl, typically 2-benzimidazolyl, benzoxazolyl, typically 2- benzoxazolyl, or quinazolyl, typically 2-quinazolinyl. Appropriate partially or, especially, completely saturated analogous radicals may also be considered, such as 2-tetrahydrofuryl, 2- or 3-pyrrolidinyl, 2-, 3-, or 4-piperidyl, and also 2-or 3-morpholinyl, 2- or 3-thiomorpholinyl, 2-piperazinyl and N-mono- or N,N'-bis-lower alkyl-2-piperazinyl radicals. These radicals may also carry one or more acyclic, carbocyclic, or heterocyclic radicals, especially those mentioned hereinabove. The free valency of the heterocyclic radicals R3 or R4 must emanate from one of their carbon atoms. Heterocyclyl may be unsubstituted or substituted by one or more, preferably one or two, of the substituents named hereinbelow for R0. [0044] Heterocyclic-aliphatic radicals R3, R4, R8 or R10 especially lower alkyl radicals, especially with a maximum of 7, preferably a maximum of 4 carbon atoms, for example those named hereinabove, which carry one, two, or more heterocyclic radicals, for example those named in the preceding paragraph, the heterocyclic ring possibly being linked to the aliphatic chain also by one of its nitrogen atoms. A preferred heterocyclic-aliphatic radical R-i is, for example, imidazol-1-ylmethyl, 4-methylpiperazin-1~ylmethyl, piperazin-1-ylmethyl, 2- (morpholin-4-yl)ethyl and also pyrid-3-ylmethyl. Heterocyclyl may be unsubstituted or substituted by one or more, preferably one or two, of the substituents named hereinbelow for R0.
[0045] A heteroaliphatic radical R3, R4, R8 or Ri0 with up to 20 carbon atoms each and up to 10 heteroatoms each is an aliphatic radical which, instead of one, two, or more carbon atoms, contains identical or different heteroatoms, such as especially oxygen, sulfur, and nitrogen. An especially preferred arrangement of a heteroaliphatic radical Ri takes the form of oxa-alkyl radicals in which one or more carbon atoms are replaced in a preferably linear alkyl by oxygen atoms preferably separated from one another by several (especially 2) carbon atoms so that they form a repeating group, if need be multi-repeating group (O-CH2-CH2-)q, wherein q = 1 to 7.
[0046] Especially preferred as R3, R4, R8 or Ri0, apart from acyl, is lower alkyl, particlularly methyl or ethyl; lower alkoxycarbonyl-lower alkyl, especially methoxycarbonylmethyl or 2-(tert-butoxycarbonyl)ethyl; carboxy-lower alkyl, especially carboxymethyl or 2-carboxyethyl; or cyano-lower alkyl, especially 2-cyanoethyl. [0047] An acyl radical R3, R4, R6, R7, Rs, R9, or R10with up to 30 carbon atoms derives from a carboxylic acid, functionally modified if need be, an organic sulfonic acid, or a phosphoric acid, such as pyro- or orthophosphoric acid, esterified if need be. [0048] An acyl designated Ac1 and derived from a carboxylic acid, functionally modified if need be, is especially one of the subformula Y-C(=W)-, wherein W is oxygen, sulfur, or imino and Y is hydrogen, hydrocarbyl R0 with up to 29 carbon atoms, hydrocar- byloxy R°-O-, an amino group or a substituted amino group, especially one of the formula R0HN- or R0R0N- (wherein the R0 radicals may be identical or different from one another). [0049] The hydrocarbyl (hydrocarbon radical) R0 is an acyclic (aliphatic), carbocyclic, or carbocyclic-acyclic hydrocarbon radical, with up to 29 carbon atoms each, especially up to 18, and preferably up to 12 carbon atoms, and is saturated or unsaturated, unsubstituted or substituted. Instead of one, two, or more carbon atoms, it may contain identical or different heteroatoms, such as especially oxygen, sulfur, and nitrogen in the acyclic and/or cyclic part; in the latter case, it is described as a heterocyclic radical (heterocyclyl radical) or a hetero- cyclic-acyclic radical.
[0050] Unsaturated radicals are those, which contain one or more, especially conjugated and/or isolated, multiple bonds (double or triple bonds). The term cyclic radicals includes also aromatic and non-aromatic radicals with conjugated double bonds, for example those wherein at least one 6-member carbocyclic or a 5- to 8-member heterocyclic ring contains the maximum number of non-cumulative double bonds. Carbocyclic radicals, wherein at least one ring is present as a 6-member aromatic ring (i.e. a benzene ring), are defined as aryl radicals.
[0051] An acyclic unsubstituted hydrocarbon radical R0 is especially a straight- chained or branched lower alkyl-, lower alkenyl-, lower alkadienyl-, or lower alkinyl radical. Lower alkyl R0 is, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, and also n-pentyl, isopentyl, n-hexyl, isohexyl and n-heptyl; lower alkenyl is, for example, allyl, propenyl, isopropenyl, 2- or 3-methallyl and 2- or 3-butenyl; lower alkadienyl is, for example, 1-penta-2,4-dienyl; lower alkinyl is, for example, propargyl or 2-butinyl. In corresponding unsaturated radicals, the double bond is especially located in a position higher than the α-position in relation to the free valency.
[0052] A carbocyclic hydrocarbon radical R0 is especially a mono-, bi-, or polycyclic cycloalkyl-, cycloalkenyl-, or cycloalkadienyl radical, or a corresponding aryl radical. Preference is for radicals with a maximum of 14, especially 12, ring-carbon atoms and 3- to 8-, preferably 5- to 7-, and most especially 6-member rings which can also carry one or more, for example two, acyclic radicals, for example those named above, especially the lower alkyl radicals, or other carbocyclic radicals. Carbocyclic-acyclic radicals are those in which an acyclic radical, especially one with a maximum of 7, preferably a maximum of 4 carbon atoms, such as especially methyl, ethyl and vinyl, carries one or more carbocyclic, if need be aromatic radicals of the above definition. Special mention is made of cycloalkyl- lower and aryl-lower alkyl radicals, as well as their analogues which are unsaturated in the ring and/or chain, and which carry the ring at the terminal carbon atom of the chain. [0053] Cycloalkyl R0 has most especially from 3 up to and including 10 carbon atoms and is, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, as well as bicyclo[2,2,2]octyl, 2-bicyclo[2,2,1]heptyl, and adamantyl, which may also be substituted by 1 , 2, or more, for example lower, alkyl radicals, especially methyl radicals; cycloalkenyl is for example one of the monocyclic cycloalkyl radicals already named which carries a double bond in the 1-, 2-, or 3 position. Cycloalkyl-lower alkyl or -lower alkenyl is for example a -methyl, -1- or -2-ethyl, -1- or -2-vinyl, -1-, -2-, or -3-propyl or -allyl substituted by one of the above-named cycloalkyl radicals, those substituted at the end of the linear chain being preferred.
[0054] An aryl radical R0 is most especially a phenyl, also a naphthyl, such as 1- or
2-naphthyl, a biphenylyl, such as especially 4-biphenylyl, and also an anthryl, fluorenyl and azulenyl, as well as their aromatic analogues with one or more saturated rings. Preferred aryl-lower alkyl and -lower alkenyl radicals are, for example, phenyl-lower alkyl or phenyl- lower alkenyl with a terminal phenyl radical, such as for example benzyl, phenethyl, 1-, 2-, or 3-phenylpropyl, diphenylmethyl (benzhydryl), trityl, and cinnamyl, and also 1- or 2-naphthyl- methyl. Aryl may be unsubstituted or substituted. [0055] Heterocyclic radicals, including heterocyclic-acyclic radicals, are especially monocyclic, but also bi- or polycyclic, aza-, thia-, oxa-, thiaza-, oxaza-, diaza-, triaza-, or tetrazacyclic radicals of an aromatic character, as well as corresponding heterocyclic radicals of this type which are partly or most especially wholly saturated; if need be, for example as in the case of the above-mentioned carbocyclic or aryl radicals, these radicals may carry further acyclic, carbocyclic, or heterocyclic radicals and/or may be mono-, di-, or polysubstituted by functional groups. The acyclic part in heterocyclic-acyclic radicals has for example the meaning indicated for the corresponding carbocyclic-acyclic radicals. Most especially they are unsubstituted or substituted monocyclic radicals with a nitrogen, oxygen, or sulfur atom, such as 2-aziridinyl, and especially aromatic radicals of this type, such as pyrrolyl, for example 2-pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3-, or 4-pyridyl, and also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl; analogous bicyclic radicals with an oxygen, sulfur, or nitrogen atom are, for example, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, or benzothienyl, typically 2- or 3- benzothienyl; preferred monocyclic and bicyclic radicals with several heteroatoms are, for example, imidazolyl, typically 2-imidazolyl, pyrimidinyl, typically 2-or 4-pyrimidinyl, oxazolyl, typically 2-oxazolyl, isoxazolyl, typically 3-isoxazolyl, or thiazolyl, typically 2-thiazolyl, and benzimidazolyl, typically 2-benzimidazolyl, benzoxazolyl, typically 2-benzoxazolyl, or quinazolyl, typically 2-quinazolinyl. Appropriate partially or, especially, completely saturated analogous radicals may also be considered, such as 2-tetrahydrofuryl, 4-tetrahydrofuryl, 2- or 3-pyrrolidyl, 2-, 3-, or 4-piperidyl, and also 2-or 3-morpholinyl, 2- or 3-thiomorpholinyl, 2- piperazinyl, and N,N'-bis-lower alkyl-2-piperazinyl radicals. These radicals may also carry one or more acyclic, carbocyclic, or heterocyclic radicals, especially those mentioned hereinabove. Heterocyclic-acyclic radicals are especially derived from acyclic radicals with a maximum of 7, preferably a maximum of 4 carbon atoms, for example those named hereinabove, and may carry one, two, or more heterocyclic radicals, for example those named hereinabove, the ring possibly being linked to the aliphatic chain also by one of its nitrogen atoms.
[0056] As already mentioned, a hydrocarbyl (including a heterocyclyl) may be substituted by one, two, or more identical or different substituents (functional groups); one or more of the following substituents may be considered: lower alkyl; free, etherified and esterified hydroxyl groups; carboxy groups and esterified carboxy groups; mercapto- and lower alkylthio- and, if need be, substituted phenylthio groups; halogen atoms, typically chlorine and fluorine, but also bromine and iodine; halogen-lower alky) groups; oxo groups which are present in the form of formyl (i.e. aldehydo) and keto groups, also as corresponding acetals or ketals; azido groups; nitro groups; cyano groups; primary, secondary and preferably tertiary amino groups, amino-lower alkyl, mono- or disubstituted amino-lower alkyl, primary or secondary amino groups protected by conventional protecting groups (especially lower alkoxycarbonyl, typically tert-butoxycarbony!) lower alkylenedioxy, and also free or functionally modified sulfo groups, typically sulfamoyl or sulfo groups present in free form or as salts. The hydrocarbyl radical may also carry carbamoyl, ureido, or guanidino groups, which are free or which carry one or two substituents, and cyano groups. The above use of the word "groups" is taken to imply also an individual group. [0057] Halogen-lower alkyl contains preferably 1 to 3 halogen atoms; preferred is trifluoromethyl or chloromethyl.
[0058] An etherified hydroxyl group present in the hydrocarbyl as substituent is, for example, a lower alkoxy group, typically the methoxy-, ethoxy-, propoxy-, isopropoxy-, butoxy-, and tert-butoxy group, which may also be substituted, especially by (i) heterocyclyl, whereby heterocyclyl can have preferably 4 to 12 ring atoms, may be unsaturated, or partially or wholly saturated, is mono- or bicyclic, and may contain up to three heteroatoms selected from nitrogen, oxygen, and sulfur, and is most especially pyrrolyl, for example 2- pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, and also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, benzothienyl, typically 2- or 3-benzothienyl; imidazolyl, typically 1- or 2-imidazolyl, pyrimidinyl, typically 2-or 4-pyrimidinyl, oxazolyl, typically 2- oxazolyl, isoxazolyl, typically 3-isoxazolyl, thiazolyl, typically 2-thiazolyl, benzimidazolyl, typically 2-benzimidazolyl, benzoxazolyl, typically 2-benzoxazolyl, quinazolyl, typically 2- quinazolinyl, 2-tetrahydrofuryl, 4-tetrahydrofuryl, 2- or 4-tetrahydropyranyl, 1-, 2- or 3- pyrrolidyl, 1-, 2-, 3-, or 4-piperidyl, 1-, 2-or 3-morpholinyl, 2- or 3-thiomorpholinyl, 2- piperazinyl or N,N'-bis-lower alkyl-2-piperazinyl; and also (ii) by halogen atoms, for example mono-, di-, or polysubstituted especially in the 2-position, as in the 2,2,2-trichloroethoxy, 2- chloroethoxy, or 2-iodoethoxy radical, or (iii) by hydroxy or (iv) lower alkoxy radicals, each preferably monosubstituted, especially in the 2-position, as in the 2-methoxyethoxy radical. Such etherified hydroxyl groups are also unsubstituted or substituted phenoxy radicals and phenyl-lower alkoxy radicals, such as especially benzyloxy, benzhydryloxy, and triphenylmethoxy (trityloxy), as well as heterocyclyloxy radicals, wherein heterocyclyl can have preferably 4 to 12 ring atoms, may be unsaturated, or partially or wholly saturated, is mono- or bicyclic, and may contain up to three heteroatoms selected from nitrogen, oxygen, and sulfur, and is most especially pyrrolyl, for example 2-pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, and also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, benzothienyl, typically 2- or 3-benzothienyl; imidazolyl, typically 1- or 2-imidazolyl, pyrimidinyl, typically 2- or 4-pyrimidinyl, oxazolyl, typically 2-oxazolyl, isoxazolyl, typically 3- isoxazolyl, thiazolyl, typically 2-thiazolyl, benzimidazolyl, typically 2-benzimidazolyl, benz- oxazolyl, typically 2-benzoxazolyl, quinazolyl, typically 2-quinazolinyl, 2-tetrahydrofuryl, 4- tetrahydrofuryl, 2- or 4-tetrahydropyranyl, 1-, 2- or 3-pyrrolidyl, 1-, 2-, 3-, or 4-piperidyl, 1-, 2- or 3-morpholinyl, 2- or 3-thiomorpholinyl, 2-piperazinyl or N,N'-bis-lower alkyl-2-piperazinyl; such as especially 2- or 4-tetrahydropyranyloxy.
[0059] Etherified hydroxyl groups in this context are taken to include silylated hydroxy! groups, typically for example tri-lower alkylsilyloxy, typically tri methyls ilyloxy and dimethyl-tert-butylsilyloxy, or phenyldi-lower alkylsilyloxy and lower alkyl-diphenylsilyloxy. [0060] An esterified hydroxyl group present in the hydrocarbyl as a substituent is, for example, lower alkanoyloxy.
[0061] A carboxyl group present in the hydrocarbyl as a substituent is one in which the hydrogen atom is replaced by one of the hydrogen radicals characterised hereinabove, preferably a lower alkyl- or phenyl-lower alkyl radical; an example of an esterified carboxyl group is lower alkoxycarbonyl or phenyl-lower alkoxycarbonyl substituted if need be in the phenyl part, especially the methoxy, ethoxy, tert-butoxy, and benzyloxycarbonyl group, as well as a lactonised carboxyl group.
[0062] A primary amino group -NH2 as substituent of the hydrocarbyls may also be present in a form protected by a conventional protecting group. A secondary amino group carries, instead of one of the two hydrogen atoms, a hydrocarbyl radical, preferably an unsubstituted one, typically one of the above-named, especially lower alkyl, and may also be present in protected form.
[0063] A tertiary amino group present in the hydrocarbyl as substituent carries 2 different or, preferably, identical hydrocarbyl radicals (including the heterocyclic radicals), such as the unsubstituted hydrocarbyl radicals characterised hereinabove, especially lower alkyl. [0064] A preferred amino group is one with the formula R1I(Ri2)N-, wherein R11 and
R12 are independently in each case hydrogen, unsubstituted acyclic CrC7-hydrocarbyl (such as especially C-i-C4alkyl or C2-C4alkenyl) or monocyclic aryl, aralkyl, or aralkenyl, substituted if necessary by CrC4-alkyl, CrC4-alkoxy, halogen, and/or nitro, and having a maximum of 10 carbon atoms, where the carbon-containing radicals may be interlinked through a carbon- carbon bond or an oxygen atom, a sulfur atom, or a nitrogen atom substituted if necessary by hydrocarbyl. In such a case, they form a nitrogen-containing heterocyclic ring with the nitrogen atom of the amino group. The following are examples of especially preferred disubstituted amino groups: di-lower alkylamino, typically dimethylamino or diethylamino, pyrrolidino, imidazol-1-yl, piperidino, piperazino, 4-lower alkylpiperazino, morpholino, thiomorpholino and piperazino or 4-methylpiperazino, as well as diphenylamino and dibenzylamino substituted if need be, especially in the phenyl part, for example by lower- alkyl, lower-alkoxy, halogen, and/or nitro; of the protected groups, especially lower alkoxy- carbonylamino, typically tert-butoxycarbonylamino, phenyl-lower alkoxycarbonylamino, typically 4-methoxybenzyloxycarbonylamino, and 9-fluorenylmethoxycarbonylamino. [0065] Amino-lower alkyl is most especially substituted in the 1 -position of the lower alkyl chain by amino and is especially aminomethyl.
[0066] Mono- or disubstituted- amino-lower alkyl is amino-lower alkyl substituted by one or two radicals, wherein amino-lower alkyl is most especially substituted by amino in the 1 -position of the lower alkyl chain and is especially aminomethyl; the amino substituents here are preferably (if 2 substituents are present in the respective amino group independently of one another) from the group comprising lower alkyl, such as especially methyl, ethyl or n-propyl, hydroxy-lower alkyl, typically 2-hydroxyethyl, C3-C8cycloalkyl, especially cyclohexyl, amino-lower alkyl, typically 3-aminopropyl or 4-aminobutyl, N-mono- or N,N-di(lower alkyl)-amino-lower alkyl, typically 3-(N,N-dimethylamino)propyl, amino, N-mono- or N,N-di-lower alkylamino and N-mono- or N,N-di-(hydroxy-lower alkyl)amino. [0067] Disubstituted amino-lower alkyl is also a 5 or 6-membered, saturated or unsaturated heterocyclyl bonded to lower alkyl via a nitrogen atom (preferably in the 1- position) and having 0 to 2, especially 0 or 1 , other heteroatoms selected from oxygen, nitrogen, and sulfur, which is unsubstituted or substituted, especially by one or two radicals from the group comprising lower alkyl, typically methyl, and also oxo. Preferred here is pyrrolidino (1-pyrrolidinyl), piperidino (1-piperidinyl), piperazino (1-piperazinyl), 4-lower alkylpiperazino, typically 4-methylpiperazino, imidazolino (1-imidazolyl), morpholino (4-morpho- linyl), or also thiomorpholino, S-oxo-thiomorpholino, or S,S~dioxothiomorpholino. [0068] Lower alkylenedioxy is especially methylenedioxy.
[0069] A carbamoyl group carrying one or two substituents is especially amino- carbonyl (carbamoyl) which is substitiuted by one or two radicals at the nitrogen; the amino substituents here are preferably (if 2 substituents are present in the respective amino group independently of one another) from the group comprising lower alkyl, such as especially methyl, ethyl or n-propyl, hydroxy-lower alkyl, typically 2-hydroxyethyl, C3-C8cycloalkyl, especially cyclohexyl, amino-lower afkyl, typically 3-aminopropyl or 4-aminobutyl, N-mono- or N,N-di(lower alkyl)-amino-lower alkyl, typically 3-(N,N-dimethylamino)propyl, amino, N-mono- or N,N-di-lower alkylamino and N-mono- or N,N-di-(hydroxy-lower alkyl)amino; disubstituted amino in aminocarbamoyl is also a 5 or 6-membered, saturated or unsaturated heterocyclyl with a bonding nitrogen atom and 0 to 2, especially 0 or 1 , other heteroatoms selected from oxygen, nitrogen, and sulfur, which is unsubstituted or substituted, especially by one or two radicals from the group comprising lower alkyl, typically methyl, and also oxo. Preferred here is pyrrolidino (1-pyrrolidinyl), piperidino (1-piperidinyl), piperazino (1-piperazinyl), 4-lower al- kylpiperazino, typically 4-methylpiperazino, imidazolino (1-imidazolyl), morpholino (4-morpho- linyl), or also thiomorpholino, S-oxo-thiomorpholino, or S,S-dioxothiomorpholino. [0070] An acyl derived from an organic sulfonic acid, which is designated Ac2, is especially one with the subformula R°-SO2-, wherein R0 is a hydrocarbyl as defined above in the general and specific meanings, the latter also being generally preferred here. Especially preferred is lower alkylphenylsulfonyl, especially 4-toluenesulfonyl. [0071] An acyl derived from a phosphoric acid, esterified if necessary, which is designated Ac3 , is especially one with the subformula R°O(R°O)P(=O)-, wherein the radicals R0 are, independently of one another, as defined in the general and specific meanings indicated above.
[0072] Reduced data on substituents given hereinbefore and hereinafter are considered to be preferences.
[0073] Preferred compounds according to the invention are, for example, those wherein R0 has the following preferred meanings: lower alkyl, especially methyl or ethyl, amino-lower alkyl, wherein the amino group is unprotected or is protected by a conventional amino protecting group - especially by lower alkoxycarbonyl, typically tert-lower alkoxy- carbonyl, for example tert-butoxycarbonyl - e.g. aminomethyl, R1S-, R- or preferably S-1- aminoethyl, tert-butoxycarbonylaminomethyl or R1S-, R-, or preferably S-1-(tert-butoxy- carbonylamino)ethyl, carboxy-lower alkyl, typically 2-carboxyethyl, lower alkoxycarbonyl- lower alkyl, typically 2-(tert-butoxycarbonyl)ethyl, cyano-lower alkyl, typically 2-cyanoethyl, tetrahydropyranyloxy-lower alkyl, typically 4-(tetrahydropyranyl)-oxymethyl, morpholino-lower alky], typically 2-(morpholino)ethyl, phenyl, lower alkylphenyl, typically 4-methylphenyl, lower alkoxyphenyl, typically 4-methoxyphenyl, imidazolyl-lower alkoxyphenyl, typically 4-[2- (imidazol-1-yl)ethyl)oyxphenyl, carboxyphenyl, typically 4-carboxyphenyl, lower alkoxy- carbonylphenyl, typically 4-ethoxycarbonylphenyl or 4-methoxyphenyl, halogen-lower alkylphenyl, typically 4-chloromethylphenyl, pyrrolidinophenyl, typically 4-pyrrolidinophenyl, imidazol-1-ylphenyl, typically 4-(imidazolyl-1-yl)phenyl, piperazinophenyl, typically 4-piperazi- nophenyl, (4-lower alkylpiperazino)phenyl, typically 4-(4-methylpiperazino)phenyl, morpholinophenyl, typically 4-morpholinophenyl, pyrrolidino-lower alkylphenyl, typically A- pyrrolidinomethylphenyl, imidazol-1-yl-lower alkylphenyl, typically 4-(imidazolyl-1-yl- methyl)phenyl, piperazino-lower alkylphenyl, typically 4-piperazinomethylphenyl, (4-lower alkylpiperazinomethyO-phenyl, typically 4-(4-methylpiperazinomethyl)phenyl, morpholino- lower alkylphenyl, typically 4-morpholinomethylphenyl, piperazinocarbonylphenyl, typically A- piperazinocarbonylphenyl, or (4-lower alkyl-piperazino)phenyl, typically 4-(4-methylpiper- azino)phenyl.
[0074] Preferred acyl radicals Ac1 are acyl radicals of a carboxylic acid which are characterised by the subformula R°-CO-, wherein R0 has one of the above general and preferred meanings of the hydrocarbyl radical R0. Especially preferred radicals R0 here are lower alkyl, especially methyl or ethyl, amino-lower alkyl, wherein the amino group is unprotected or protected by a conventional amino protecting group, especially by lower al- koxycarbonyl, typically tert-lower alkoxycarbonyl, for example tert-butoxycarbonyl, e.g. aminomethyl, R1S-, R-, or preferably S-1-aminoethyl, tert-butoxycarbonylaminomethyl or R1S-, R-, or preferably S-1-(tert-butoxycarbonylamino)ethyl, carboxy-lower alkyl, typically 2- carboxyethyl, lower alkoxycarbonyl-lower alkyl, typically 2-(tert-butoxycarbonyl)ethyl, tetrahydropyranyloxy-lower alkyl, typically 4-(tetrahydropyranyl)oxymethyl, phenyl, imidazolyl- lower alkoxyphenyl, typically 4-[2-(imidazol-1-yl)ethyl]oyxphenyl, carboxyphenyl, typically 4- carboxyphenyl, lower alkoxycarbonylphenyl, typically 4-ethoxycarbonylphenyl, halogen-lower alkylphenyl, typically 4-chloromethylphenyl, imidazol-1-ylphenyl, typically 4-(imidazolyl-1-yl)- phenyl, pyrrolidino-lower alkylphenyl, typically 4-pyrrolidinomethylphenyl, piperazino-lower alkylphenyl, typically 4-piperazinomethylphenyl, (4-lower alkylpiperazinomethyl)phenyl, typically 4-(4-methylpiperazinomethyl)phenyl, morpholino-lower alkylphenyl, typically A- morpholinomethylphenyl, piperazinocarbonylphenyl, typically 4-piperazinocarbonylphenyl, or (4-lower alkylpiperazino)phenyl, typically 4-(4-methylpiperazino)phenyl. [0075] A further preferred Acyl Ac1 is derived from monoesters of carbonic acid and is characterised by the subformula R°-O-CO-. The lower alkyl radicals, especially tert-butyl, are especially preferred hydrocarbyl radicals R0 in these derivatives. [0076] Another preferred Acyl Ac1 is derived from amides of carbonic acid (or also thiocarbonic acid) and is characterised by the formula R°HN-C(=W)- or R°R°N-C(=W)-, wherein the radicals R0 are, independently of one another, as defined above and W is sulfur and especially oxygen. In particular, compounds are preferred wherein Ac1 is a radical of formula R°HN-C(=W)-, wherein W is oxygen and R0 has one of the following preferred meanings: morpholino-lower alkyl, typically 2-morpholinoethyl, phenyl, lower alkoxyphenyl, typically 4-methoxyphenyl or 4-ethoxyphenyl, carboxyphenyl, typically 4-carboxyphenyl, or lower alkoxycarbonylphenyl, typically 4-ethoxycarbonylphenyl.
[0077] A preferred acyl Ac2 of subformula R°-SO2-, wherein R0 is a hydrocarbyl as defined in the above general and specific meanings, is lower alkylphenylsulfonyl, typically 4- toluenesulfonyl.
[0078] If p is 0, the nitrogen atom bonding R3 is uncharged. If p is 1 , then R4 must also be present, and the nitrogen atom bonding R3 and R4 (quaternary nitrogen) is then positively charged.
[0079] The definitions for an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms each, or for a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms each and up to 9 heteroatoms each, or acyl with up to 30 carbon atoms each, preferably match the definitions given for the corresponding radicals R3 and R4. Especially preferred is R5 lower alkyl, especially methyl, or most especially hydrogen. [0080] Z is especially lower alkyl, most especially methyl or hydrogen.
[0081] If the two bonds indicated by wavy lines are missing in ring A, then no double bonds (tetra-hydrogenated derivatives) are present between the carbon atoms characterised in formula I by the numbers 1 , 2, 3, and 4, but only single bonds, whereas ring B is aromatic (double bonds between the carbon atoms characterised in formula I by 8 and 9 and those characterised by 10 and 11). If the two bonds indicated by wavy lines are missing in ring B1 then no double bonds (tetra-hydrogenated derivatives) are present between the carbon atoms characterised in formula I by the numbers 8, 9, 10, and 11 , but only single bonds, whereas ring A is aromatic (double bonds between the carbon atoms characterised in formula I by 1 and 2 and those characterised by 3 and 4). If the total of four bonds indicated by wavy lines are missing in rings A and B, and are replaced by a total of 8 hydrogen atoms, then no double bonds (octa-hydrogenated derivatives) are present between the carbon atoms numbered 1 , 2, 3, 4, 8, 9, 10, and 11 in formula I, but only single bonds. [0082] By their nature, the compounds of the invention may also be present in the form of pharmaceutically, i.e. physiologically, acceptable salts, provided they contain salt- forming groups. For isolation and purification, pharmaceutically unacceptable salts may also be used. For therapeutic use, only pharmaceutically acceptable salts are used, and these salts are preferred.
[0083] Thus, compounds of formula I having free acid groups, for example a free sulfo, phosphoryl or carboxyl group, may exist as a salt, preferably as a physiologically acceptable salt with a salt-forming basic component. These may be primarily metal or ammonium salts, such as alkali metal or alkaline earth metal salts, for example sodium, potassium, magnesium or calcium salts, or ammonium salts with ammonia or suitable organic amines, especially tertiary monoamines and heterocyclic bases, for example triethylamine, tri-(2-hydroxyethyl)-amine, N-ethylpiperidine or N.N'-dimethylpiperazine. [0084] Compounds of the invention having a basic character may also exist as addition salts, especially as acid addition salts with inorganic and organic acids, but also as quaternary salts. Thus, for example, compounds which have a basic group, such as an amino group, as a substituent may form acid addition salts with common acids. Suitable acids are, for example, hydrohalic acids, e.g. hydrochloric and hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid or perchloric acid, or aliphatic, alicyclic, aromatic or heterocyclic carboxylic or sulfonic acids, such as formic, acetic, propionic, succinic, glycolic, lactic, malic, tartaric, citric, fumaric, maleic, hydroxymaleic, oxalic, pyruvic, phenylacetic, benzoic, p-aminobenzoic, anthranilic, p-hydroxybenzoic, salicylic, p-aminosalicylic acid, pamoic acid, methanesulfonic, ethanesulfonic, hydroxyethanesulfonic, ethylenedisulfonic, halobenzenesulfonic, toluenesulfonic, naphthalenesulfonic acids or sulfanilic acid, and also methionine, tryptophan, lysine or arginine, as well as ascorbic acid. [0085] In view of the close relationship between the novel compounds (especially of formula I) in free form and in the form of their salts, including those salts that can be used as intermediates, for example in the purification or identification of the novel compounds, and of their solvates, any reference hereinbefore and hereinafter to the free compounds is to be understood as referring also to the corresponding salts, and the solvates thereof, for example hydrates, as appropriate and expedient.
[0086] The compounds of formula A, B, C, D, I, II, III, IV, V or Vl especially those wherein R5 is hydrogen, possess valuable pharmacological properties. [0087] In the case of the groups of radicals or compounds mentioned hereinbefore and hereinafter, genera! definitions may, insofar as appropriate and expedient, be replaced by the more specific definitions stated hereinbefore and hereinafter.
[0088] Preference is given to a compounds of formula I, II, III, IV, V, Vl wherein
R-I and R2 independently of each other are lower alkyl, lower alkyl substituted by halogen, C6-C14aryl, hydroxy, lower alkoxy, phenyl-lower alkoxy, phenyloxy, lower alkanoyloxy, benzoyloxy, amino, lower alkylamino, lower alkanoylamino, phenyl-lower alkylamino, N,N-di-lower alkylamino, N,N-di-(phenyl-lower alkyl)amino, cyano, mercapto, lower alkylthio, carboxy, lower alkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, N,N-di- lower alkylcarbamoyl, sulfo, lower alkanesulfonyl, lower alkoxysulfonyl, aminosulfonyl, N- lower alkylaminosulfonyl or N,N-di-lower alkylaminosulfonyl; halogen; lower alkoxy; C6-Ci4aryloxy; C6-Ci4aryl-lower alkoxy; lower alkanoyloxy; C6-C14arylcarbonyloxy; amino monosubstituted or disubstituted by lower alkyl, C6-C14aryl, C6-Ci4aryl-lower alkyl, lower alkanoyl or
Figure imgf000024_0001
cyano; nitro; mercapto; lower alkylthio; C6-C14arylthio; C6-Ci4aryl-lower alkylthio; lower alkanoylthio; C6-C14aryl-lower alkanoylthio; carboxy; lower alkoxycarbonyl, C6-C14aryl-lower alkoxycarbonyl; C6-C14aryloxycarbonyl; carbamoyl; carbamoyl N-mono- or N,N-disubstituted by lower alkyl, C6-Ci4aryl or C6-C14aryl-lower alkyl; sulfo; C6-C14arylsulfonyl; C6-Ci4aryl-lower alkanesulfonyl; lower -alkanesulfonyl; or aminosulfonyl N-mono- or N,N-disubstituted by lower alkyl, C6-Ci4aryl or C6-C14aryl-lower alkyl, wherein C6-C14aryl is an aryl radical with 6 to 12 carbon atoms in the ring system, which may be unsubstituted or substituted by halogen, phenyl or naphthyl, hydroxy, lower alkoxy, phenyl-lower alkoxy, phenyloxy, lower alkanoyloxy, benzoyloxy, amino, lower alkylamino, lower alkanoylamino, phenyl-lower alkylamino, N, N-di-lower alkylamino, N,N-di-(phenyl-lower alkyl)amino, cyano, mercapto, lower alkylthio, carboxy, lower alkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, N, N-di- lower alkylcarbamoyl, sulfo, lower alkanesulfonyl, lower alkoxysulfonyl, aminosulfonyl, N- lower alkylaminosulfonyl or N, N-di-lower alkylaminosulfonyl; n and m are independently of each other 0 or 1 or 2, preferably 0;
R3> R4, Re, RIQ are independently of each other hydrogen, lower alkyl, lower alkenyl or lower alkadienyl, which are each unsubstituted or monosubstituted or polysubsituted, preferably monosubstituted or disubstituted by a substituent independently selected from lower alkyl; hydroxy; lower alkoxy, which may be unsubstituted or mono-, di-, or trisubstituted by (i) heterocyclyl with 4 to 12 ring atoms, which may be unsaturated, wholly saturated, or partly saturated, is monocyclic or bicyclic and may contain up to three heteroatoms selected from nitrogen, oxygen and sulfur, and is most especially pyrrolyl, for example 2-pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, or in a broader sense also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, benzothienyl, typically 2- or 3-benzothienyl; imidazolyl, typically 1- or 2-imidazolyl, pyrimidinyl, typically 2-or 4-pyrimidinyl, oxazolyl, typically 2-oxazolyl, isoxazolyl, typically 3-isoxazolyl, thiazolyl, typically 2-thiazolyl, benzimidazolyl, typically 2-benzimidazolyl, benzoxazolyl, typically 2-benzoxazolyl, quinazolyl, typically 2-quinazolinyl, 2- tetrahydrofuryl, 4-tetrahydrofuryl, 4-tetrahydropyranyl, 1-, 2- or 3-pyrrolidyl, 1-, 2-, 3-, or 4-piperidyl, 1-, 2-or 3-morpholinyl, 2- or 3-thiomorpholinyl, 2-piperazinyl or N,N'-bis-lower alkyl-2-piperazinyl, (ii) by halogen, (iii) by hydroxy or (iv) by lower alkoxy; phenoxy; phenyl-lower alkoxy; heterocyclyloxy, wherein heterocyclyl is pyrrolyl, for example 2- pyrrolyl or 3-pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, or in a broader sense also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl, indolyl, typically 2- or 3- indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, benzothienyl, typically 2- or 3-benzothienyl; imidazolyl, typically 1- or 2-imidazolyl, pyrimidinyl, typically 2-or 4- pyrimidinyl, oxazolyl, typically 2-oxazolyl, isoxazolyl, typically 3-isoxazolyl, thiazolyl, typically 2-thiazolyl, benzimidazolyl, typically 2-benzimidazolyl, benzoxazolyl, typically 2- benzoxazolyl, quinazolyl, typically 2-quinazolinyl, 2-tetrahydrofuryl, 4-tetrahydrofuryl, 2- or 4-tetrahydropyranyl, 1-, 2- or 3-pyrrolidyl, 1-, 2-, 3-, or 4-piperidyl, 1-, 2-or 3- morpholinyl, 2- or 3-thiomorpholinyl, 2-piperazinyl or N,N'-bis-lower alkyl-2-piperazinyl, such as especially 2- or 4-tetrahydropyranyloxy; lower alkanoyloxy; carboxy; lower alkoxycarbonyl; phenyl-lower alkoxycarbonyl; mercapto; lower alkylthio; phenylthio; halogen; halogen-lower alkyl; oxo (except in the 1 -position, because otherwise acyl); azido; nitro; cyano; amino; mono-lower alkylamino; di-lower alkylamino; pyrrolidino; imidazol-1-yl; piperidino; piperazino; 4-lower alkylpiperazino; morpholino; thiomorpholino; diphenylamino or dibenzylamino unsubstituted or substituted in the phenyl part by lower alkyl, lower alkoxy, halogen and/or nitro; lower alkoxycarbonylamino; phenyl-lower alkoxycarbonylamino unsubstituted or substituted in the phenyl part by lower alkyl or lower alkoxy; fluorenylmethoxycarbonylamino; amino-lower alkyl; monosubstituted or disubstituted amino-lower alkyl, wherein the amino substituent is selected from lower alkyl, hydroxy-lower alkyl, C3-C8cycloalkyl, amino-lower alkyl, N-mono- or N,N-di(-lower -alkyl)amino-fower alkyl, amino, N-mono- or N,N-di-lower alkylamino and N-mono- or N,N-di-(hydroxy-lower alkyl)amino; pyrrolidino-lower alkyl; piperidino-lower alkyl; piperazino-lower alkyl; 4-lower alkylpiperazino-lower alkyl; imidazol-1-yl-lower alkyl; morpholino-lower alkyl; thiomorpholino-lower alkyl; S-oxo-thiomorpholino-lower alkyl; S,S-dioxothiomorpholino-lower alkyl; lower alkylendioxy; sulfamoyl; sulfo; carbamoyl; ureido; guanidino; cyano; aminocarbonyl (carbamoyl) and aminocarbonyloxy, which are substituted by one or two radicals on the nitrogen, wherein the amino substituents are selected independently of one another from the group comprising lower alkyl, hydroxy- lower alkyl, C3-C8cycloalkyl, amino-lower alkyl, N-mono- or N,N-di(-lower alkyl)amino- lower alkyl, amino, N-mono- or N,N-di-lower alkylamino and N-mono- or N,N-di-(hydroxy- lower alkyl)amino; pyrrolidinocarbonyl; piperidinocarbonyl; piperazinocarbonyl; 4-lower alkylpiperazinocarbonyl; imidazolinocarbonyl; morpholinocarbonyl; thio- morpholinocarbonyl; S-oxo-thiomorpholinocarbonyl; and S,S-dioxothiomorpholino; phenyl, naphthyl, phenyl-lower alkyl or phenyl-lower alkenyl with a terminal phenyl radical, which is unsubstituted or monosubstituted or disubstituted by the radicals named above as substituents of lower alkyl, lower alkenyl or lower alkadienyl; or heterocyclyl-lower alkyl, wherein heterocyclyl is pyrrolyl, for example 2-pyrrolyl or 3- pyrrolyl, pyridyl, for example 2-, 3- or 4-pyridyl, or in a broader sense also thienyl, for example 2- or 3-thienyl, or furyl, for example 2-furyl, indolyl, typically 2- or 3-indolyl, quinolyl, typically 2- or 4-quinolyl, isoquinolyl, typically 3- or 5-isoquinolyl, benzofuranyl, typically 2-benzofuranyl, chromenyl, typically 3-chromenyl, benzothienyl, typically 2- or 3- benzothienyl; imidazolyl, typically 1- or 2-imidazolyl, pyrimidinyl, typically 2-or 4- pyrimidinyl, oxazolyl, typically 2-oxazolyl, isoxazolyl, typically 3-isoxazolyl, thiazolyl, typically 2-thiazolyl, benzimidazolyl, typically 2-benzimidazolyl, benzoxazolyl, typically 2- benzoxazolyl, quinazolyl, typically 2-quinazolinyl, 2-tetrahydrofuryl, 4-tetrahydrofuryl, 2- or 4-tetrahydropyranyl, 1-, 2- or 3-pyrrolidyl, 1-, 2-, 3-, or 4-piperidyl, 1-, 2-or 3- morpholinyl, 2- or 3-thiomorpholinyl, 2-piperazinyl or N,N'-bis-lower alkyl-2-piperazinyl, which in each case are unsubstituted or monosubstituted or disubstituted by the radicals named above as substituents of lower alkyl, lower alkenyl, or lower alkadienyl; or acyl of the subformula Y-C(=W)-, wherein W is oxygen and Y is hydrogen, R0, R°-O-, R0HN-, or R0R0N- (wherein the radicals R0 may be the same or different), or acyl of the subformula R°-SO2-, whereby R4 may also be absent for the compound of formula II; or
R4 is absent for compounds of formula II, hydrogen or CH3 for compounds of formula I, and
R3 is acyl of the subformula Y-C(=W)-, wherein W is oxygen and Y Is hydrogen, R0, R°-O-, R0HN-, or R0R0N- (wherein the radicals R0 may be the same or different), or is acyl of the subformula R°-SO2-, wherein R0 in the said radicals has the following meanings: substituted or unsubstituted lower alkyl, especially methyl or ethyl, amino-lower alkyl hydroxy-lower alkyl, wherein the amino group is unprotected or is protected by a conventional amino protecting group - especially by lower alkoxycarbonyl, typically tert-lower alkoxycarbonyl, for example tert- butoxycarbonyl - e.g. aminomethyl, R1S-, R- or preferably S-1-aminoethyl, tert- butoxycarbonylaminomethyl or R1S-, R-, or preferably S-i-(tert-butoxycarbonylamino) ethyl, carboxy-lower alkyl, typically 2-carboxyethyl, lower alkoxycarbonyl-lower alkyl, typically 2-(tert-butoxycarbonyl)ethyl, cyano-lower alkyl, typically 2-cyanoethyl, tetrahydropyranyloxy-lower alkyl, typically 4-(tetrahydropyranyl) oxymethyl, morpholino- lower alkyl, typically 2-(morpholino)ethyl, phenyl, lower alkylphenyl, typically 4- methylphenyl, lower alkoxyphenyl, typically 4-methoxyphenyl, imidazolyl-lower alkoxyphenyl, typically 4-[2-(imidazol-1-yl)ethyl) oxyphenyl, carboxyphenyl, typically 4- carboxyphenyl, lower alkoxycarbonylphenyl, typically 4-ethoxycarbonylphenyl or 4- methoxyphenyl, halogen-lower alkylphenyl, typically 4-chloromethylphenyl, pyrrolidino- phenyl, typically 4-pyrrolidinophenyl, imidazol-1-ylphenyl, typically 4-(imidazolyl-1- yl)phenyl, piperazinophenyl, typically 4-piperazinophenyl, (4-lower alkylpiperazino)phenyl, typically 4-(4-methylpiperazino)phenyl, morpholinophenyl, typically 4-morpholinophenyl, pyrrolidino-lower alkylphenyl, typically 4-pyrrolidinomethylphenyl, imidazol-1-yl-lower alkylphenyl, typically 4-(imidazolyl-1-ylmethyl)phenyl, piperazino-lower alkylphenyl, typically 4-piperazinomethylphenyl, (4-lower alkylpiperazinomethyl)-phenyl, typically 4-(4- methylpiperazinomethyl)phenyl, morpholino-lower alkylphenyl, typically 4- morpholinomethylphenyl, piperazinocarbonylphenyl, typically 4-piperazinocarbonylphenyl, or (4-lower alkylpiperazino)phenyl, typically 4-(4-methylpiperazino)phenyl. p is 0 if R4 is absent, or is 1 if R3 and R4 are both present and in each case are one of the aforementioned radicals (for compounds of formula II);
R5 is hydrogen or lower alkyl, especially hydrogen, X stands for 2 hydrogen atoms, for O, or for 1 hydrogen atom and hydroxy; or for 1 hydrogen atom and lower alkoxy;
Z is hydrogen or especially lower alkyl, most especially methyl; and for compounds for formula II, either the two bonds characterised by wavy lines are preferably absent in ring A and replaced by 4 hydrogen atoms, and the two wavy lines in ring B each, together with the respective parallel bond, signify a double bond; or also the two bonds characterised by wavy lines are absent in ring B and replaced by a total of 4 hydrogen atoms, and the two wavy lines in ring A each, together with the respective parallel bond, signify a double bond; or both in ring A and in ring B all of the 4 wavy bonds are absent and are replaced by a total of 8 hydrogen atoms; or a salt thereof, if at least one salt-forming group is present.
[0089] Particular preference is given to a compound of formula I wherein; m and n are each 0;
R3 and R4 are independently of each other: hydrogen, lower alkyl unsubstituted or mono- or disubstituted, especially monosubstituted, by radicals selected independently of one another from carboxy; lower alkoxycarbonyl; and cyano; or
R4 is hydrogen or -CH3, and R3 is as defined above or preferably R3 is, acyl of the subformula R°-CO, wherein R0 is lower alkyl; amino-lower alkyl, wherein the amino group is present in unprotected form or is protected by lower alkoxycarbonyl; tetrahydropyranyloxy-lower alkyl; phenyl; imidazolyl-lower alkoxyphenyl; carboxyphenyl; lower alkoxycarbonylphenyl; halogen-lower alkylphenyl; imidazol-1-ylphenyl; pyrrolidino-lower alkylphenyl; piperazino-lower alkylphenyl; (4-lower alkylpiperazinomethyl)phenyl; morpholino-lower alkylphenyl; piperazinocarbonylphenyl; or (4-lower alkylpiperazino)phenyl; or is acyl of the subformula R°-O-CO-, wherein R0 is lower alkyl; or is acyl of the subformula R°HN-C(=W)-, wherein W is oxygen and R0 has the following meanings: morpholino-lower alkyl, phenyl, lower alkoxyphenyl, carboxyphenyl, or lower alkoxycarbonylphenyl; or R3 is lower alkylphenylsulfonyl, typically 4-toluenesulfonyl; wherein further specific examples of preferred R3 groups are described below for the preferred compounds of formula II, R5 is hydrogen or lower alkyl, especially hydrogen, X stands for 2 hydrogen atoms or for O; Z is methyl or hydrogen; or a salt thereof, if at least one salt-forming group is present.
[0090] Particular preference is given to a compound of formula Il wherein m and n are each 0;
R3 and R4 are independently of each other hydrogen, lower alkyl unsubstituted or mono- or disubstituted, especially monosubstituted, by radicals selected independently of one another from carboxy; lower alkoxycarbonyl; and cyano; whereby R4 may also be absent; or
R4 is absent, and
R3 is acyl from the subformula R°-CO, wherein R0 is lower alkyl, especially methyl or ethyl; amino-lower alkyl, wherein the amino group is unprotected or protected by lower alkoxycarbonyl, typically tert-lower alkoxycarbonyl, for example tert-butoxycarbonyl, e.g. aminomethyl, R1S-, R-, or preferably S-1-aminoethyl, tert-butoxycarbonylaminomethyl or R1S-, R-, or preferably S-1-(tert-butoxycarbonylamino)ethyl; tetrahydropyranyloxy-lower alkyl, typically 4-(tetrahydropyranyl)oxymethyl; phenyl; imidazolyl-lower alkoxyphenyl, typically 4-[2-(imidazol-1-yl)ethyl)oyxphenyl; carboxyphenyl, typically 4-carboxyphenyl; lower alkoxycarbonylphenyl, typically 4-methoxy- or 4-ethoxycarbonylphenyl; halogen- lower alkylphenyl, typically 4-chloromethylphenyl; imidazol-1-ylphenyl, typically 4- (imidazolyl-i-yl)phenyl; pyrrolidino-lower alkylphenyl, typically 4-pyrrolidinomethylphenyl; piperazino-lower alkylphenyl, typically 4-piperazinomethylphenyl; (4-lower alkylpiperazinomethyl)phenyl, typically 4-(4-methylpiperazinomethyl)phenyl; morpholino- lower alkylphenyl, typically 4-morpholinomethylphenyl; piperazinocarbonylphenyl, typically 4-piperazinocarbonylphenyl; or (4-lower alkylpiperazino)phenyl, typically 4-(4- methylpiperazino)phenyl; or is acyl of the subformula R°-O-CO-, wherein R0 is lower alkyl; or is acyl of the subformula R°HN-C(=W)-, wherein W is oxygen and R0 has the following preferred meanings: morpholino-lower alkyl, typically 2-morpholinoethyf, phenyl, lower alkoxyphenyl, typically 4-methoxyphenyl or 4-ethoxyphenyl, carboxyphenyl, typically 4-carboxyphenyl, or lower alkoxycarbonylphenyl, typically 4-ethoxycarbonylphenyl; or is lower alkylphenylsulfonyl, typically 4-toluenesulfonyl; p is 0 if R4 is absent, or is 1 if R3 and R4 are both present and in each case are one of the aforementioned radicals;
R5 is hydrogen or lower alkyl, especially hydrogen, X stands for 2 hydrogen atoms or for O; Z is methyl or hydrogen; and either the two bonds characterised by wavy lines are preferably absent in ring A and replaced by 4 hydrogen atoms, and the two wavy lines in ring B each, together with the respective parallel bond, signify a double bond; or also the two bonds characterised by wavy lines are absent in ring B and replaced by a total of 4 hydrogen atoms, and the two wavy lines in ring A each, together with the respective parallel bond, signify a double bond; or both in ring A and in ring B all of the 4 wavy bonds are absent and are replaced by a total of 8 hydrogen atoms; or a salt thereof, if at least one salt-forming group is present.
[0091] Most especially preferred compounds of formula Il are selected from;
8,9,10,11-Tetrahydrostaurosporine; N-[4-(4-methylpiperaziN-1-ylmethyl)benzoyl]-1 ,2,3,4- tetrahydrostaurosporine; N-(4-chloromethylbenzoyl)-1 ,2,3,4-tetrahydrostaurosporine; N-(4- (pyrrolidin-1-ylmethyl)benzoyl)-1 ,2,3,4-tetrahydrostaurosporine; N-(4-(morpholin-4- ylmethyl)benzoyl)-1 ,2,3,4-tetrahydrostaurosporine; N-(4-(piperazin-1 -ylmethyl)benzoyl)- 1 ,2,3,4-tetrahydrostaurosporine; N-ethyl-1 ,2,3,4-tetrahydrostaurosporine; N-tosyl-1 ,2,3,4- tetrahydrostaurosporine; N-triflouroacetyl-1 ,2,3,4-tetrahydrostaurosporine; N-[4-(2-imidazol- 1 -yl-ethoxy)benzoy!]-1 ,2,3,4-tetrahydrostaurosporine; N-methoxycarbonylmethyl-1 ,2,3,4- tetrahydrostaurosporine; N-carboxymethyl-1 ,2,3,4-tetrahydrostaurosporine; N- terephthaloylmethyl ester-1 ,2,3,4-tetrahydrostaurosporine; N-terephthaloyl-1 ,2,3,4- tetrahydrostaurosporine; N-(4-ethylpiperazinylcarbonylbenzoyl)-1 ,2,3,4- tetrahydrostaurosporine; N-(2-cyanoethyl)-1 ,2,3,4-tetrahydrostaurosporine; N-benzoyl- 1 ,2,3,4-tetrahydrostaurosporine; N,N-dimethyl -1 ,2,3,4-tetrahydrostaurosporinium iodide; N- BOC-glycyl-1 ,2,3,4-tetrahydrostaurosporine; N-glycyl-1,2,3,4-tetrahydrostaurosporine; N-(3- (tert-butoxycarbonyl)propyl)-1 ,2,3,4-tetrahydrostaurosporine; N-(3-carboxypropyl)-1 ,2,3,4- tetrahydrostaurosporine; N-(4-imidazol-1-yl)benzoyl]-1 ,2,3,4-tetrahydrostaurosporine; N- [(tetrahydro-2h-pyran-4-yloxy)acetyl]-1 ,2,3,4-tetrahydrostaurosporine; N-BOC-l-alanyl- 1 ,2,3,4-tetrahydrostaurosporine; N-l-alanyl-1 ,2,3,4-tetrahydrostaurosporine hydrochloride; N-methyl-1 ,2,3,4-tetrahydro-6-methylstaurosporine; N-(4-carboxyphenylaminocarbonyl)- 1 ,2,3,4-tetrahydrostaurosporine; N-(4-ethylphenylaminocarbonyl)-1 ,2,3,4- tetrahydrostaurosporine; N-(N-phenylaminocarbonyl)-1 ,2,3,4-tetrahydrostaurosporine; N- (N-[2-(1-morpholino)ethyl]aminocarbonyl)-1 ,2,3,4-tetrahydrostaurosporine; N-(N~[4- methoxyphenyl]aminocarbonyl)-1 ,2,3,4-tetrahydrostaurosporine; 1 ,2,3,4-tetrahydro-6- methylstaurosporine; N-BOC-1 ,2,3,4-tetrahydrostaurosporine; N-BOC-1 ,2,3,4-tetrahydro-6- methylstaurosporine; N-BOC-1 , 2,3,4-tetrahydro-6-methyl-7-oxo-staurosporine; 1 ,2,3,4,8,9,10,11-octahydrostaurosporine; or a pharmaceutically acceptable salt thereof, if at least one salt-forming group is present.
[0092] Most especially preferred is the compound of formula I designated 1 ,2,3,4- tetrahydrostaurosporine, or a (particularly pharmaceutically acceptable) salt thereof (here, m und n in formula I are 0, R3 is hydrogen, R4 is absent, provided no salt is present (p = 0), or is hydrogen if a salt is present (p = 1), R5 is hydrogen, the two bonds represented by wavy lines are absent in Ring A and are replaced by a total of 4 hydrogen atoms and the two bonds represented by wavy lines in Ring B are in each case a double bond together with the parallel bonds, X stands for 2 hydrogen atoms, and Z is methyl). [0093] Most especially preferred are the compounds of formula A wherein;
A) X= O; R1 , R2, R5 = H; Q= -(CH2)2-O-CH(CH2)OH-(CH2)2- (LY 333531 )
B) X= O; R1, R2, R5 = H; Q= -(CH2)2-O-CH(CH2N(CH3)2)-(CH2)2-
C) X= 2 hydrogen atoms; R1, R2, R5 = H; Q= H-V y>c"*
°H (MLR52 ; CAS=I 55416-34-5)
[0094] Most especially preferred are the compounds of formula I wherein;
A) X= 2 hydrogen atoms; Ri,R2, R3, R5 = H; R4= CH3; Z=CH3 (staurosporine)
B) X= 1 hydrogen and 1 hydroxy atoms in (R) or (S) isomeric form; Ri,R2, R31R5 = H; R4= CH3; Z=CH3 (UCN-01 and UCN-02)
C) X= 2 hydrogen atoms; Ri,R2, R5 = H; R4= CH3; R3,= benzoyl; Z=CH3 (CGP41251 or PKC412 or MIDOSTAURIN) D) X= O; R11R2, R5 = H; R3,= CH3; R4= ethyloxycarbonyl; Z=CH3 (NA 382 ; CAS= 143086-33- 3)
E) X= 1 hydrogen and 1 hydroxy atom; R1, R2, Rs = H; R3= CH3; Z=CH3; and R4 is selected from -(CH2)2OH; -CH2CH(OH)CH2OH; -CO(CH2)2CO2Na; -(CH2)3CO2H; - COCH2N(CHa)2;
!-COCH2-N N-CH3
Figure imgf000032_0001
F) X= 2 hydrogen atoms; R1, R2, R5 = H; R3= CH3; Z=CH3; and R4 is selected from N-[O- (tetrahydropyran-4-yl )-D-lactoyl]; N-[2-methyl-2-(tetrahydropyran-4-yloxy)-propionyl; N- [0-(tetrahydropyran-4-yl )-L-lactoyl]; N-[0-(tetrahydropyran-4-yl )-D-lactoyl]; N-[2- (tetrahydro-pyran-4-yloxy)-acetyi)]
G) X=O; R1, R2, R5 = H; R3= CH3; Z=CH3; and R4 is selected from N-[0-(tetrahydropyran-4-yl )-D-lactoyl]; N-[2-(tetrahydro-pyran-4-yloxy)-acetyl)]
H) X=1 hydrogen and 1 hydroxy atom ; R1, R2, R5 = H; R3= CH3; Z=CH3; and R4 is selected from N-[0-(tetrahydropyran-4-yl )-D-lactoyl]; N-[2-(tetrahydro-pyran-4-yloxy)-acetyl)] The abbreviation "CAS" means the CHEMICAL ABSTRACTS registry number. [0095] The most preferred compounds of formula I e.g. MIDOSTAURIN
[International Nonproprietary Name] are covered and have been specifically described by the European patent No. 0 296 110 published on December 21, 1988, as well as in US patent No. 5;093,330 published on March 3, 1992, and Japanese Patent No. 2 708 047. Other preferred compounds are covered and described by the patent applications WO 95/32974 and WO 95/32976 both published on December 7, 1995. All the compounds described in these documents are incorporated into the present application by reference. [0096] Most especially preferred are the compounds of formula III wherein;
A) X= 2 hydrogen atoms; R11R2, R5 = H; R6= CH3; R7= methyloxycarbonyl; Z=H (2- methyl K252a)
B) X= 2 hydrogen atoms; R11R2, R5, R6 = H; R7= methyloxycarbonyl; Z= H (K-252a)
C) X= 2 hydrogen atoms; R11R2, R5, R6 = H; R7= methyloxycarbonyl; Z= CH3 (KT-5720) [0097] Most especially preferred are the compounds of formula IV wherein;
A) X= O; R1, R2, R5 = H; R9= CH2-NMe2; R8= CH3 ; m'=n'=2
B) X= O; R1, R2, R5 = H; R9= CH2-NH2; R8= CH3 ; m'=2; n'=1 (Ro-31-8425; CAS=151342- 35-7)
[0098] Most especially preferred are the compounds of formula V wherein;
A) X= O; R1, R2, R5 = H; R
Figure imgf000032_0002
8= CH3; -(CH2)3-NH2; (Ro-31-7549; CAS=I 38516-31) B) X= O; R1, R21 R5 = H; R8= CH3; R10= -(CH2)3-S-(C=NH)-NH2; (Ro-31-8220 ; CAS=125314-64-9))
C)
Figure imgf000033_0001
CH3; R1CF -CH3;
[0099] Most especially preferred are the compounds of formula Vl wherein;
A) X= 2 hydrogen atoms; R11R2, R5 = H; R4= CH3; Z=CH3 ; R3 is selected from methyl or (C1-
C10)alkyl, arylmethyl, C6H2CH2- .
[00100] STAUROSPORINE DERIVATIVES and their manufacturing process have been specifically described in many prior art documents, well known by the man skilled in the art.
[00101] Compounds of formula A, B, C, D and their manufacturing process have for instance, been described in the European patents No. 0 657 458 published on June 14, 1995, in the European patents No. 0 624 586 published on November 17, 1994, in the European patents No. 0 470 490 published on February 12, 1992, in the European patents No. 0 328 026 published on August 16, 1989, in the European patents No. 0 384 349 published on August 29, 1990, as well as in many publications such as Barry M. Trost* and Weiping Tang Org. Lett, 3(21), 3409-3411.
[00102] Compounds of formula I and their manufacturing process has been specifically described in the European patents No. 0 296 110 published on December 21 , 1988, as well as in US patent No. 5;093,330 published on March 3, 1992, and Japanese Patent No. 2 708 047. Compounds of formula I having a tetrahydropyran-4-yl )-lactoyl substitution on R4 have been described in the European patent No. 0 624 590 published on November 17, 1994. Other compounds have been described in the European patent No. 0 575 955 published December 29, 1993, European patent No. 0 238 011 published on September 23, 1987 ( UCN-01), International patent application EP98/04141 published as WO99/02532 on July 03, 1998.
[00103] Compounds of formula Il and their manufacturing process has been specifically described in the European patents No. 0 296 110 published on December 21, 1988, as well as in US patent No. 5;093,330 published on March 3, 1992, and Japanese Patent No. 2 708 047.
[00104] Compounds of formula III and their manufacturing process has been specifically described in the patent applications claiming the priority of the US patent application US 920102 filed on July 24, 1992. (i.e., European patents No. 0 768 312 published on April 16, 1997, No. 1 002 534 published May 24, 2000, No. 0 651 754 published on May 10, 1995.
[00105] Compounds of formula IV and their manufacturing process has been specifically described in the patent applications claiming the priority of the British patent applications GB 9309602 and GB 9403249 respectively filed on May 10, 1993, and on February 21 , 1994. (i.e., European patents No. 0 624 586 published on November 17, 1994, No. 1 002 534 published May 24, 2000, No. 0 651 754 published on May 10, 1995. [00106] Compounds of formula V and their manufacturing process has been specifically described in the patent applications claiming the priority of the British patent applications GB 8803048, GB 8827565, GB 8904161 and GB 8928210 respectively filed on February 10, 1988, November 25, 1988, February 23, 1989 and December 13, 1989. (i.e., European patents No. 0 328 026 published on August 16, 1989, and No. 0 384 349 published August 29, 1990).
[00107] Compounds of formula Vl and their manufacturing process has been specifically described in the patent applications claiming the priority of the US patent applications 07/777,395 (Con), filed on October 10, 1991 (i.e., International patent application WO 93/07153 published on April 15, 1993).
[00108] In each case where citations of patent applications or scientific publications are given in particular for the STAUROSPORINE DERIVATIVE compounds, the subject- matter of the final products, the pharmaceutical preparations and the claims is hereby incorporated into the present application by reference to this publications. [00109] The structure of the active agents identified by code nos., generic or trade names may be taken from the actual edition of the standard compendium "The Merck Index" or from databases, e.g., Patents International (e.g., IMS World Publications). The corresponding content thereof is hereby incorporated by reference.
[00110] The preferred STAUROSPORINE DERIVATIVE according to the invention is
N-[{9S, 10R, 11 R, 13R)-2,3, 10, 11 ,12,13-hexahydro-10-methoxy-9-methyl-1 -oxo-9, 13-epoxy- IH.gH-diindoloti ^.S-ghiS'^M'-lmlpyrrolotS^-JllUlbenzodiazonin-H-y^-Λ/- methylbenzamide of the formula (VII):
Figure imgf000035_0001
or a salt thereof, (hereinafter: "Compound of formula VII or M I DOSTAU Rl N").
[00111] Compound of formula VII is also known as MIDOSTAURIN [International
Nonproprietary Name] or PKC412.
[00112] MIDOSTAURIN is a derivative of the naturally occurring alkaloid staurosporine, and has been specifically described in the European patent No. 0 296 110 published on December 21 , 1988, as well as in US patent No. 5;093,330 published on March
3, 1992, and Japanese Patent No. 2 708 047.
[00113] It has now surprisingly been found that MIDOSTAURIN possesses therapeutic properties, which render it particularly useful as a modulator of transformed cells containing at least one SPTBN 1-FLT3 translocation, and especially in the treatment and prophylaxis of myeloid leukemias, especially atypical chronic myeloid leukemia. This compound shows an unexpected high potency toward treating cancer patients harboring a
SPTBN 1-FLT3 translocation, which appears to be an activating mutation, as shown in Ba/F3 cell lines harbouring the SPTBN 1-FLT3 translocation construct.
[00114] STAUROSPORINE DERIVATIVES e.g. MIDOSTAURIN were originally identified as inhibitor of protein kinase C (PKC) (Meyer T, Regenass U, Fabbro D, et al: lnt J
Cancer 43: 851-856, 1989).
[00115] It has now surprisingly been found that STAUROSPORINE DERIVATIVES possess therapeutic properties that render it particularly useful as an inhibitor of SPTBN1-
FLT3 translocation-mediated signalling pathways and especially in the treatment and prophylaxis of diseases mediated by SPTBN1-FLT3 translocation-associated signalling pathways, especially atypical chronic myeloid leukemia. This compound shows an unexpected high potency toward cell lines harboring SPTBN1-FLT3 translocation, an activating mutation especially in atypical chronic myeloid leukemia.
[00116] The present invention thus concerns the use of STAUROSPORINE
DERIVATIVES for the preparation of a drug for inhibiting SPTBN 1-FLT3 translocation protein product and downstream effects (e.g., mediated by FGFR3, ras, c-fos and/or TGF- beta).
[00117] The present invention more particularly concerns the use of
STAUROSPORINE DERIVATIVES for the preparation of a drug for the treatment of diseases involving SPTBN 1 or Flt3 signalling pathways or SPTBN 1-FLT3 translocation- modulated pathways or SPTBN1-FLT3 translocation gene product. Preferred diseases are myeloid leukemias, including especially atypical chronic myeloid leukemias.
[00118] The present invention more particularly concerns the use of
STAUROSPORINE DERIVATIVES for the preparation of a drug for the treatment of myeloid leukemias, including especially atypical chronic myeloid leukemias.
[00119] In still another embodiment, the instant invention provides a method for treating diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product comprising administering to a mammal in need of such treatment a therapeutically effective amount of one or more STAUROSPORINE
DERIVATIVES, or a pharmaceutically acceptable salts or prodrugs thereof.
[00120] Preferably the instant invention provides a method for treating mammals especially humans suffering from diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product comprising administering to a mammal in need of such treatment a FLT3 receptor tyrosine kinase activity inhibiting amount of Λ/-[(9S, 10R, 11 R, 13R)-2,3, 10,11 ,12,13-hexahydro-10-methoxy-9- methyl-1-oxo-9J3-epoxy-1Hf9H-diindolo[1 ,2>3-gh:3\2\1I-lm]pyrrolo[3,4-j][1 ,7]benzodiazonin-
11-yl]-Λ/-methylbenzamide of the formula (VII).
[00121] The instant invention also concerns a method wherein the therapeutically effective amount of the compound of formula VII is administered to a mammal subject 7 to 4 times a week or about 100 % to about 50% of the days in the time period, for a period of from one to six weeks, followed by a period of one to three weeks, wherein the agent is not administered and this cycle being repeated for from 1 to several cycles. [00122] Preferably, this method is used for treating myeloid leukemia, including especially atypical chronic myeloid leukemia.
[00123] In another embodiment, the instant invention relates to the use of
STAUROSPORINE DERIVATIVES for the preparation of a pharmaceutical composition for use in treating diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product.
[00124] STAUROSPORINE DERIVATIVES have useful pharmacological properties.
In particular, it inhibits the activity of FLT3 and / or SPTBN1-FLT3 translocation gene product in concentrations in the range of 0.01 to 50 μM.
[00125] In vivo, the activity of the STAUROSPORINE DERIVATIVES especially compounds of formula I or II, can be demonstrated, for example, in a single oral administration per day to animals at doses in the range of 5 to 300 or 100 to 200 mg/kg of body weight per day.
[00126] The STAUROSPORINE DERIVATIVES are therefore very highly suitable for the treatment of diseases, which respond to inhibition of the FLT3 or TGF-beta signalling pathways and / or SPTBN1-FLT3 translocation gene product transcription, e.g., atypical chronic myeloid leukemia.
[00127] In the present description, the term "treatment" includes both prophylactic or preventative treatment as well as curative or disease suppressive treatment, including treatment of patients at risk of contracting the disease or suspected to have contracted the disease as well as ill patients. This term further includes the treatment for the delay of progression of the disease.
[00128] The term "curative" as used herein means efficacy in treating ongoing episodes involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or
SPTBN1-FLT3 translocation gene product.
[00129] The term "prophylactic" means the prevention of the onset or recurrence of diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or
SPTBN 1-FLT3 translocation gene product, including the polypeptide encoded by a mutated gene comprising a SPTBN 1-FLT3 translocation.
[00130] The term "delay of progression" as used herein means administration of the active compound to patients being in a pre-stage or in an early phase of the disease to be treated, in which patients for example a pre-form of the corresponding disease is diagnosed or which patients are in a condition, e.g. during a medical treatment or a condition resulting from an accident, under which it is likely that a corresponding disease will develop.
[00131] To demonstrate that STAUROSPORINE DERIVATIVES are particularly suitable for the treatment of atypical chronic myeloid leukemia with good therapeutic margin and other advantages, clinical trials can be carried out in a manner known to the skilled person.
[00132] The precise dosage of STAUROSPORINE DERIVATIVES to be employed for inhibiting SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1- FLT3 translocation gene product transcription depends upon several factors including the host, the nature and the severity of the condition being treated, the mode of administration. However, in general, satisfactory inhibition of SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product is achieved when the STAUROSPORINE DERIVATIVES is administered (check preferred administration) parenterally, e.g., intraperitoneally, intravenously, intramuscularly, subcutaneously, intratumorally, or rectally, or enterally, e.g., orally, preferably intravenously or orally, intravenously at a daily dosage of 1-300 mg/kg body weight or, for most larger primates, a daily dosage of 50-5000, preferably 500-3000 mg, in human trials a total dose of 225 mg/day was most presumably the Maximum Tolerated Dose (MTD). A preferred intravenous daily dosage is 1-75 mg/kg body weight or, for most larger primates, a daily dosage of 50-1500 mg. A typical intravenous dosage is 20 mg/kg, three to five times a week. [00133] Most preferably, the STAUROSPORINE DERIVATIVES, especially
MlDOST AURlN, are administered orally, by dosage forms such as microemulsions, soft gels or solid dispersions in dosages up to 150 mg/day, administered in one, two or three times. [00134] Usually, a small dose is administered initially and the dosage is gradually increased until the optimal dosage for the host under treatment is determined. The upper limit of dosage is that imposed by side effects and can be determined by trial for the host being treated.
[00135] The STAUROSPORINE DERIVATIVES may be combined with one or more pharmaceutically acceptable carriers and, optionally, one or more other conventional pharmaceutical adjuvants and administered enterally, e.g. orally, in the form of tablets, capsules, caplets, etc. or parenterally, e.g., intraperitoneally or intravenously, in the form of sterile injectable solutions or suspensions. The enteral and parenteral compositions may be prepared by conventional means. [00136] The infusion solutions according to the present invention are preferably sterile. This may be readily accomplished, e.g. by filtration through sterile filtration membranes. Aseptic formation of any composition in liquid form, the aseptic filling of vials and/or combining a pharmaceutical composition of the present invention with a suitable diluent under aseptic conditions are well known to the skilled addressee. [00137] The STAUROSPORINE DERIVATIVES may be formulated into enteral and parenteral pharmaceutical compositions containing an amount of the active substance that is effective for inhibiting SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product, such compositions in unit dosage form and such compositions comprising a pharmaceutically acceptable carrier.
[00138] The STAUROSPORINE DERIVATIVES can be used alone or combined with at least one other pharmaceutically active compound for use in these pathologies. These active compounds can be combined in the same pharmaceutical preparation or in the form of combined preparations "kit of parts" in the sense that the combination partners can be dosed independently or by use of different fixed combinations with distinguished amounts of the combination partners, i.e., simultaneously or at different time points. The parts of the kit of parts can then, e.g., be administered simultaneously or chronologically staggered, that is at different time points and with equal or different time intervals for any part of the kit of parts. Non-limiting examples of compounds which can be cited for use in combination with STAUROSPORINE DERIVATIVES are cytotoxic chemotherapy drugs, such as cytosine arabinoside, daunorubicin, doxorubicin, cyclophosphamide, VP-16, etc. Further, STAUROSPORINE DERIVATIVES could be combined with other inhibitors of signal transduction or other oncogene-targeted drugs with the expectation that significant synergy would result.
[00139] Examples of useful compositions are described in the European patents No. 0
296 110, No. 0 657 164, No. 0 296 110, No.O 733 372, No.O 711 556, No.O 711 557. [00140] The preferred compositions are described in the European patent No. 0 657
164 published on June 14, 1995. The described pharmaceutical compositions comprise a solution or dispersion of compounds of formula I such as MIDOSTAURIN in a saturated polyalkylene glycol glyceride, in which the glycol glyceride is a mixture of glyceryl and polyethylene glycol esters of one or more C8-C18 saturated fatty acids. [00141] Two manufacture processes of such compositions are described hereafter.
Composition A: [00142] Gelucire 44/14 (82 parts) is melted by heating to 60 DEG C. Powdered
MIDOSTAURIN (18 parts) is added to the molten material. The resulting mixture is homogenised and the dispersion obtained is introduced into hard gelatin capsules of different size, so that some contain a 25mg dosage and others a 75mg dosage of the MIDOSTAURIN. The resulting capsules are suitable for oral administration.
Composition B:
[00143] Gelucire 44/14 (86 parts) is melted by heating to 60 DEG C. Powdered
MIDOSTAURIN (14 parts) is added to the molten material. The mixture is homogenised and the dispersion obtained is introduced into hard gelatin capsules of different size, so that some contain a 25mg dosage and others a 75mg dosage of the MIDOSTAURIN. The resulting capsules are suitable for oral administration.
[00144] Gelucire 44/14 available commercially from Gattefosse; is a mixture of esters of C8-C18 saturated fatty acids with glycerol and a polyethylene glycol having a molecular weight of about 1500, the specifications for the composition of the fatty acid component being, by weight, 4-10% capryllc acid, 3-9% capric acid, 40-50% lauric acid, 14-24% myristic acid, 4-14% palmitic acid and 5-15% stearic acid.
[00145] A preferred example of Gelucire formulation consists of:
Gelucire (44/14): 47 g
MIDOSTAURIN: 3.0gfilled into a 60 mL Twist off flask [00146] A preferred example of soft gel will contain the following Microemulsion:
Cornoil glycerides 85.0 mg
Polyethylenglykol 400 128.25 mg
Cremophor RH 40 213.75 mg
MIDOSTAURIN 25.0 mg
DL alpha Tocopherol 0.5 mg
Ethanol absolute 33.9 mg
Total 486.4 mg
[00147] However, it should be clearly understood that it is for purposes of illustration only.
[00148] In a preferred embodiment this invention relates to use or method as described herein, wherein the daily effective amount of the compound of formula VII, is 100 to 300 mg, preferably 125 mg to 250 mg most preferably 220 to 230 mg, preferably 225 mg. [00149] Most preferably the compound of formula VII, is administered once, two or three times a day, for a total dose of 100 to 300 mg daily.
[00150] In a very preferred embodiment the compound of formula VII, is administered three times a day, for a total dose of 220 to 230 preferably 225 mg daily, and preferably at a dose per administration of 70 to 80 mg, preferably 75 mg.
[00151] In still another embodiment, this invention relates to an article of manufacture comprising packaging material, and Λ/-[(9S,10/?,11R13R)-2,3,10,11 ,12,13-hexahydro-10- methoxy-9-methyl-1 -oxo-9, 13-epoxy-1 W,9/-/-diindolo[1 ^.S-gtrS1^1, 1 '-lm]pyrrolo[3,4- j][1,7]benzodiazonin-11-yl]-/V-methylbenzamide of the formula (VII) or a pharmaceutically acceptable salts thereof, contained within said packaging material, wherein said packaging material comprises label directions which indicate that said compound of formula (VII), or said pharmaceutically-acceptable salt, is to be administered to mammals suffering from diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product, in an amount from 50 to 500 mg, preferably 100 to 300 mg, preferably 125 mg to 250 mg, preferably 220 to 230 mg most preferably 225 mg following a specific dosage regimen to inhibit the development of diseases involv involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product.
[00152] Preferably to an article of manufacture wherein the compound of formula VII, is administered three times a day, for a total dose of 220 to 230mg, preferably 225 mg daily, and preferably a dose of 70 to 80 mg most preferably 75 mg per administration for treating myeloid leukemia, including especially atypical chronic myeloid leukemia. A preferred embodiment relates to an article of manufacture comprising softgel capsules containing 25 mg of the compound of formula VII.
[00153] The efficacy of STAUROSPORINE DERIVATIVES for the treatment of diseases involving deregulated FLT3 receptor tyrosine kinase activity is illustrated by the results of the pharmacological tests known to those skilled in the art, including those disclosed in Examples 1 and 2. These examples illustrate the invention without in any way limiting its scope.
[00154] Patient Selection for Therapy Based Upon Polymorphisms and/or Mutations.
The application of involving SPTBN 1-FLT3 translocation-associated genotypes and/or haplotypes that correlate with efficacious drug responses will be used to select patients for therapy of existing diseases. The application of genotypes and/or haplotypes that correlate with a predisposition for disease will likewise be used to select patients for preventative therapy.
[00155] A further aspect of the invention is a method for determining whether a mammal, especially a human, has a disease involving SPTBN1-FLT3 translocation, and preferably whether atypical chronic myeloid leukemia cells in such a mammal have a SPTBN 1-FLT3 translocation. Methods for detecting such SPTBN 1-FLT3 translocation include, but are not limited, to the following molecular genetic techniques: reverse- transcriptase (RT) and genomic polymerase chain reaction (PCR) using primer sequences derived from the SPTBN1-FLT3 translocation sequence encoding the SPTBN1-FLT3 fusion polypeptide (see FIG. 1 , panels 1A-1D). The forward primer sequence is preferably derived from exons 1-3 of SPTBN1 and reverse primer is preferably derived from sequences downstream from exon 12 in FLT3. In patients with this mutation the translocation genomic breakpoint can be amplified by PCR with forward and reverse primers derived from exon 3 of SPTBN 1 and FLT3 exon 13.
[00156] SPTBN1 and FLT3 rearrangement may also be identified through fluorescent in situ hybridisation (FISH), using fosmid or bacterial artificial chromosome clones (BAC) around the breakpoint of the FLT3 and SPTBN 1 genes. Suitable clones and labelling techniques will be apparent to those skilled in the art. Similarly, the SPTBN 1-FLT3 translocation may be detected at the protein level, for example by determining that the molecular size of the FLT3 is modified in the affected cells or that FLT3 protein may be isolated using antibodies, or other specific binding entities, specific to SPTBN1. [00157] A further aspect of the invention is a polynucleotide encoding a gene product comprising a fusion of the SPTBN1 and FLT3 genes, wherein the polynucleotide comprises genomic sequence of the fused SPTBN1-FLT3 translocation, and the gene product thereof, wherein the gene product is the RNA or its respective cDNA comprising the transcribed fusion gene sequence or the translated polypeptide encoded by the transcribed fusion gene sequence, and their usage as reagents in diagnostic assays for SPTBN1-FLT3 translocations and screening assays for agents capable of inhibiting the function of SPTBN 1- FLT3 gene fusion products or determining the effect of such gene products.
[00158] Hybridization Conditions
[00159] In one embodiment, an isolated nucleic acid molecule of the invention is at least 6 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the SPTBN1-FLT3 gene fusion nucleotide sequence. In another embodiment, the nucleic acid is at least 10, 25, 50, 100, 250, 500, 1000, 2000 or 2165 nucleotides in length. In another embodiment, an isolated nucleic acid molecule of the invention hybridizes to the coding region of the SPTBN1-FLT3 fusion gene. In a further embodiment, an isolated nucleic acid molecule of the invention hybridizes to the messenger RNA or corresponding cDNA. As used herein, the term "hybridizes under stringent conditions" refers to conditions for hybridization and washing under which nucleotide sequences at least 70% homologous to each other typically remain hybridized to each other. [00160] As used herein, the phrase "stringent hybridization conditions" refers to conditions under which a probe, primer or oligonucleotide will hybridize to its target sequence, but to no other sequences. Stringent conditions are sequence-dependent and selection of appropriate hybridization conditions are within the skill of one skilled in the art. Longer sequences hybridize specifically at higher temperatures than shorter sequences. Generally, stringent conditions are selected to be about 50C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50% of the probes complementary to the target sequence hybridize to the target sequence at equilibrium. Stringent conditions are known to those skilled in the art and can be found in Ausubel et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, N.Y., 6.3.1-6.3.6 (1989). Preferably, the conditions are such that sequences at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other.
[00161] Typical stringent conditions include, but are not limited to, e.g., a salt concentration of less than about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0 to 8.3 and wherein the temperature is at least about 3O0C for short probes, primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about 600C for longer probes, primers and oligonucleotides. Stringent conditions may also be achieved with the addition of destabilizing agents, such as formamide, using standard procedures known to one skilled in the art. An additional non-limiting example of stringent hybridization conditions is hybridization in a high salt buffer comprising 6xSSC, 50 mM Tris-HCI (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/ml denatured salmon sperm DNA at 650C, followed by one or more washes in 0.2xSSC, 0.01% BSA at 5O0C. Other conditions of high stringency, moderate stringency and low stringency that may be used are well-known in the art. See, e.g., Ausubel et al. (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, (1993) and Kriegler, GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY (1990).
[00162] Conservative Mutations
[00163] Naturally-occurring allelic variants of the SPTBN1 and FLT3 gene sequence may exist in the population, and may translocate to generate the SPTBN1-FLT3 fusion gene. Such changes can be introduced into the SPTBN1-FLT3 fusion gene nucleotide sequence of SEQ ID NO: 1 , on occasion leading to changes in the amino acid sequence of the encoded SPTBN1-FLT3 fusion protein, without altering the functional ability of the SPTBN1-FLT3 fusion protein. Nucleotide substitutions leading to amino acid substitutions at "non-essential" amino acid residues can be made in SEQ ID NO:1 and inserted into a plasmid or vector known in the art. A "non-essential" amino acid residue is a residue that can be altered from the sequence of SEQ ID NO:2 without altering the biological activity, whereas an "essential" amino acid residue is required for biological activity. In a specific embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence encoding a SPTBN1-FLT3 fusion protein, wherein the protein comprises an amino acid sequence at least about 85% identical, 90% identical, 95% identical, up to 99 % identical to the SPTBN 1-FLT3 fusion polypeptide sequence of SEQ ID NO:2. Preferably, the protein encoded by the nucleic acid molecule is at least about 85% identical to the SPTBN 1-FLT3 fusion polypeptide, more preferably at least about 90% homologous, and most preferably at least about 95% homologous to the given SPTBN 1-FLT3 fusion polypeptide.
[00164] Furthermore, modified SPTBN1-FLT3 fusion sequences can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of the SPTBN 1-FLT3 gene fusion, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein.
[00165] Mutations can be introduced into SEQ ID NO:1 by standard techniques, e.g., site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., K, R or H, using the one amino acid code), acidic side chains (e.g., D, E), uncharged polar side chains (e.g., F, N, Q, S, T, Y, C), nonpolar side chains (e.g., A, V, L, I, P, F, M, W), beta-branched side chains (e.g., T, V, I) and aromatic side chains (e.g., Y, F, W, H). Thus, a predicted nonessential amino acid residue in SPTBN1-FLT3 fusion polypeptide is replaced with another amino acid residue from the same side chain family. Using the one letter code for amino acids, additional contemplated conserved amino acid substitutions include, e.g., replacement of one amino acid in a group with any other amino acid listed in the same group, wherein exemplary conserved groups include but are not limited to: FYW, HY, MILV, MILF, NDEQ, NEQK1 NHQK1 QHRK and STA. [00166] Alternatively, in another embodiment, mutations can be introduced randomly along all or part of a SPTBN1-FLT3 gene fusion coding sequence, e.g., by saturation mutagenesis, and the resultant mutants can be screened for biological activity to identify mutants that retain the activity of the SPTBN1-FLT3 fusion polypeptide of SEQ ID NO:2. Following mutagenesis, the encoded modified SPTBN 1-FLT3 fusion polypeptide can be expressed by any recombinant technology known in the art and the activity of the protein can be determined.
[00167] In a related embodiment, the modified SPTBN1-FLT3 fusion polypeptide can be assayed for the ability of the modified SPTBN 1-FLT3 fusion polypeptide to transform cell lines, such as the Ba/F3 cells in the Examples.
[00168] Homology of DNA Sequences
[00169] The DNA sequence homology referred to above may be determined as the degree of identity between two sequences indicating a derivation of the first sequence from the second. The homology may suitably be determined by means of computer programs known in the art, such as GAP provided in the GCG program package. See, e.g., Needleman and Wunsch, J MoI Biol 1970 48: 443-453. Homology may also be determined by computer analysis such as, without limitations, the ClustalX computer alignment program. See, e.g., Thompson et al. Nucleic Acids Res. 1997, 25 (24): 4876-82. [00170] The term "sequence identity" refers to the degree to which two polynucleotide sequences are identical on a nucleotide-by-nucleotide basis over a particular region of comparison. The term "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over that region of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the region of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. The term "substantial identity" as used herein denotes a characteristic of a polynucleotide sequence, wherein the polynucleotide comprises a sequence that has at least 80 percent sequence identity, preferably at least 85 percent identity and often 90 to 95 percent sequence identity, more usually at least 99 percent sequence identity as compared to a reference sequence over a comparison region.
[00171] Antibodies
[00172] SPTBN 1-FLT3 polypeptides or polypeptide fragments of the invention are used to produce SPTBN1-FLT3 fusion-specific antibodies. As used herein, a " SPTBN1- FLT3 fusion-specific antibody is an antibody, e.g., a polyclonal antibody or a monoclonal antibody, that is immunoreactive to a SPTBN 1-FLT3 fusion polypeptide or polypeptide fragment, or that binds with specificity to an epitopes of a SPTBN1-FLT3 fusion polypeptides.
[00173] The preparation of polyclonal and monoclonal antibodies is well known in the art. Polyclonal antibodies may in particular be obtained as described by, e.g., Green et al.,: "Production of Polyclonal Antisera" in IMMUNOCHEMICAL PROTOCOLS (Manson, Ed.); Humana Press, 1992, pages 1-5; by Coligan et al. in CURRENT PROTOCOLS IN IMMUNOLOGY, 1992, Section 2.4.1, and by Harlow and Lane (Eds.) in "ANTIBODIES; A LABORATORY MANUAL" Cold Spring Harbor Lab. Press 1988. These protocols are hereby incorporated by reference. Monoclonal antibodies may in particular be obtained as described by, e.g., Kohler & Milstein, Nature. 1975, 256:495; Coligan et al., in CURRENT PROTOCOLS IN IMMUNOLOGY, 1992, Sections 2.5.1-2.6.7; and Harlow et al., in ANTIBODIES: A LABORATORY MANUAL; Cold Spring Harbor, Pub., 1988, page 726; which protocols are hereby incorporated by reference.
[00174] Antibodies that bind to the SPTBN 1-FLT3 fusion polypeptide can be prepared using an intact polypeptide or fragments containing small peptides of interest as the immunizing antigen. The polypeptide used to immunize an animal may be obtained by recombinant DNA techniques or by chemical synthesis, and may optionally be conjugated to a carrier protein. Commonly used carrier proteins which are chemically coupled to the peptide include keyhole limpet hemocyanin (KLH), thyroglobulin, bovine serum albumin (BSA), and tetanus toxoid. The coupled peptide may then be used to immunize the animal, which may in particular be a mouse, a rat, a hamster or a rabbit. EXAMPLE 1 :
[00175] A patient was identified with BCR-ABL negative chronic myeloid leukemia
(CML) who presented with a 46XX,t(2;13)(q3;q1),t(2;21)(p13;q11.2). Fluorescence in situ hybridization (FISH) using probes flanking the FLT3 gene at 13q12 suggested that this gene was disrupted. 5'-RACE PCR using primers to the region of FLT3 encoding the tyrosine kinase domain identified an in-frame mRNA fusion between exon 3 of SPTBN1 (spectrin, beta, non-erythrocytic 1 isoform 2, NM 178313) on chromosome 2p16 and exon 13 of FLT3 (NM 004119). Juxtaposition of SPTBN1 and FLT3 was confirmed by two color FISH and amplification of the genomic DNA breakpoint confirmed a fusion between intron 3 of SPTBN 1 and intron 12 of FLT3.
[00176] Sequences used to amplify the fusion by RT-PCR as a single step include forward primer SPTBN1 1F (5'-aga aga aga cct tea cca agt-3')(SEQ ID NO:3) and FLT3 1R (5'-tga ace agg cat get gga at-3')(SEQ ID NO:4). In patients with this mutation the translocation genomic breakpoint was amplified by PCR with forward and reverse primers derived from exon 3 of SPTBN1 (5'-cca tga cat cgt gga tgg aaa cc-3')(SEQ ID NO:5) and FLT3 exon 13 (5'-tgt ace ate tgt age tgg ctt tc-3')(SEQ ID NO:6).
[00177] The patient was initially treated with hydroxyurea and subsequently underwent an unrelated donor bone marrow transplant. She relapsed cytogenetically at 4 years but responded to donor lymphocyte infusion (DLI), achieving sustained cytogenetic and molecular (nested RT-PCR) remission. We conclude that SPTBN 1-FLT3 is a rare abnormality in BCR-ABL negative CML that is responsive to both targeted signal transduction therapy and immunotherapy by DLI.
[00178] The SPTBN1-FLT3 fusion gene is predicted to be translated into a 570 amino acid chimeric protein that retains two coiled-coil domains from SPTBN 1 and 424 amino acids from FLT3, including the entire tyrosine kinase domain. Since the t(2;'13) is readily visible by cytogenetic analysis but has not been reported previously it seems likely that SPTBN1-FLT3 is uncommon.
[00179] Forty cases were analyzed by RT-PCR to test if FLT3 might be involved more widely in BCR-ABL negative CML. Two cases were positive for the FLT3 internal tandem duplication (ITD) but mutation of residue D835 was not observed.
EXAMPLE 2:
[00180] Expression of the SPTBN1-FLT3 fusion transformed the interleukin 3 (IL-3)- dependent cell line Ba/F3 to growth factor independence and was accompanied by constitutive phosphorylation of the fusion protein and the downstream substrate ERK1/2. The growth of transformed cells was inhibited in a dose-dependent fashion by SU11657 and PKC412, but not by imatinib mesylate.
EQUIVALENTS
[00181] The present invention is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the invention. Many modifications and variations of this invention can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the invention, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present invention is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled.

Claims

Claims:
1. Use of staurosporine derivatives of formula,
Figure imgf000049_0001
wherein R1 , and R2 are, independently of one another, unsubstituted or substituted alkyl, hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl; n and m are, independently of one another, a number from and including 0 to and including 4;
R5 is hydrogen, an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms in each case, or a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms in each case, and in each case up to 9 heteroatoms, or acyl with up to 30 carbon atoms;
X stands for 2 hydrogen atoms; for 1 hydrogen atom and hydroxy; for O; or for hydrogen and lower alkoxy;
Q and Q' are independently a pharmaceutically acceptable organic bone or hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N- mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N- mono- or N,N-di-substituted aminosulfonyl; or a salt thereof, if at least one salt-forming group is present, or hydrogenated derivative thereof, for the preparation of a pharmaceutical composition for the treatment of diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product.
2. The use of a staurosporin derivative selected from the compounds of formula,
Figure imgf000050_0001
Figure imgf000050_0002
or or
Figure imgf000050_0003
wherein R1 and R2, are, independently of one another, unsubstituted or substituted alkyl, hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubsti- tuted amino, cyano, nitro, mercapto, substituted mercapto, carboxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl; n and m are, independently of one another, a number from and including zero to and including 4; n' and m' are, independently of one another, a number from and including 1 to and including 4;
R3, R4, R8 and Ri0 are, independently of one another, hydrogen, an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms in each case, a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms in each case, and in each case up to 9 heteroatoms, an acyl with up to 30 carbon atoms, wherein R4 may also be absent; or R3 is acyl with up to 30 carbon atoms and R4 not an acyl; p is 0 if R4 is absent, or is 1 if R3 and R4 are both present and in each case are one of the aforementioned radicals;
R5 is hydrogen, an aliphatic, carbocyclic, or carbocyclic-aliphatic radical with up to 29 carbon atoms in each case, or a heterocyclic or heterocyclic-aliphatic radical with up to 20 carbon atoms in each case, and in each case up to 9 heteroatoms, or acyl with up to 30 carbon atoms;
R7, R6 and R9 are acyl or -(lower alkyl) -acyl, unsubstituted or substituted alkyl, hydrogen, halogen, hydroxy, etherified or esterified hydroxy, amino, mono- or disubstituted amino, cyano, nitro, mercapto, substituted mercapto, carboxy,carbonyl, carbonyldioxy, esterified carboxy, carbamoyl, N-mono- or N,N-di-substituted carbamoyl, sulfo, substituted sulfonyl, aminosulfonyl or N-mono- or N,N-di-substituted aminosulfonyl;
X stands for 2 hydrogen atoms; for 1 hydrogen atom and hydroxy; for O; or for hydrogen and lower alkoxy;
Z stands for hydrogen or lower alkyl; and either the two bonds characterised by wavy lines are absent in ring A and replaced by 4 hydrogen atoms, and the two wavy lines in ring B each, together with the respective parallel bond, signify a double bond; or the two bonds characterised by wavy lines are absent in ring B and replaced by a total of 4 hydrogen atoms, and the two wavy lines in ring A each, together with the respective parallel bond, signify a double bond; or both in ring A and in ring B all of the 4 wavy bonds are absent and are replaced by a total of 8 hydrogen atoms; or a salt thereof, if at least one salt-forming group is present for the preparation of a pharmaceutical composition for the treatment of diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product.
3. The use of a staurosporin derivative of formula I,
Figure imgf000052_0001
wherein m and n are each 0;
R3 and R4 are independently of each other hydrogen, lower alkyl unsubstituted or mono- or disubstituted, especially monosubstituted, by radicals selected independently of one another from carboxy; lower alkoxycarbonyl; and cyano; or
R4 is hydrogen or -CH3, and R3 is acyl of the subformula R°-CO, wherein R0 is lower alkyl; amino-lower alkyl, wherein the amino group is present in unprotected form or is protected by lower alkoxycarbonyl; tetrahydropyranyloxy-lower alkyl; phenyl; imidazolyl-lower alkoxyphenyl; carboxyphenyl; lower alkoxycarbonylphenyl; halogen-lower alkylphenyl; imidazol-1-ylphenyl; pyrrolidino- lower alkylphenyl; piperazino-lower alkylphenyl; (4-lower alkylpiperazinomethyl)phenyl; morpholino-lower alkylphenyl; piperazinocarbonylphenyl; or (4-lower alkylpiperazino)phenyl; or is acyl of the subformula R°-O-CO-, wherein R0 is lower alkyl; or is acyl of the subformula R°HN-C(=W)-, wherein W is oxygen and R0 has the following meanings: morpholino-lower alkyl, phenyl, lower alkoxyphenyl, carboxyphenyl, or lower alkoxycarbonylphenyl; or R3 is lower alkylphenylsulfonyl, typically 4-toluenesulfonyl; R5 is hydrogen or lower alkyl, X stands for 2 hydrogen atoms or for O; 2 is methyl or hydrogen; or a salt thereof, if at least one salt-forming group is present for the preparation of a pharmaceutical composition for the treatment of diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN1-FLT3 translocation gene product.
4. Use according to any one of claims 1 to 3 for the treatment of atypical chronic myeloid leukemia.
5. A method for treating mammals suffering from diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product, the method comprising administering to a mammal in need of such treatment an amount of staurosporine derivatives as defined in any one of claims 1 to 3.
6. A method according to claim 5 for treating atypical chronic myeloid leukemia.
7. Use of Λ/-[(9S,10/?,11R,13/?)-2,3,10,11,12,13-hexahydro-10-methoxy-9-methyl-1- oxo-9,13-epoxy-1H,9/-/-diindolo[1 ,2,3-gh:3',2',1'-lm]pyrrolo[3,4-j][1 ,7]benzodiazonin- 11-yl]-Λ/-methylbenzamide of the formula (VII):
Figure imgf000054_0001
or a salt thereof, for the preparation of a pharmaceutical composition for the treatment of diseases involving SPTBN1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product.
8. Use according to claim 7 for the treatment of atypical chronic myeloid leukemia.
9. Pharmaceutical preparation for the treatment of atypical chronic myeloid leukemia, comprising a Λ/-[(9S, 10R, 11 R, 13R)-2,3, 10,11 ,12,13-hexahydro-10-methoxy-9-methyl- 1-oxo-9I13-epoxy-1/-/l9H-diindolo[1 ,2,3-gh:3',2l,1l-lm]pyrrolo[3,4-j][1 ,7]benzodiazonin- 11-yl]-/V-methylbenzamide of the formula (VII).
10. A method for treating mammals suffering from diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product comprising administering to a mammal in need of such treatment an inhibiting amount of Λ/-[(9S,10f?,11 R,13R)-2,3,10,11 ,12,13-hexahydro-10-methoxy-9- methyl-1-oxo-9,13-epoxy-1H,9H-diindolo[1 ,2,3-gh:3\2\1Mm]pyrrolo[3,4- j][1 ,7]benzodiazonin-11-yl]-Λ/-methylbenzamide of the formula (VII) as defined in claim 8.
11. A method according to claim 10 for treating atypical chronic myeloid leukemia.
12. A method according to any one of claims 7 to 11, wherein the therapeutically effective amount of the compound of formula VII is administered to a mammal subject 7 to 4 times a week or about 100 % to about 50% of the days in the time period, for a period of from one to six weeks, followed by a period of one to three weeks, wherein the agent is not administered and this cycle being repeated for from 1 to several cycles.
13. Use or method according to any one of claims 7 to 12, wherein the daily effective amount of the compound of formula VII, is 100 to 300 mg daily preferably 220 to 230mg, most preferably 225 mg daily.
14. Use or method according to any one of claims 7 to 13, wherein the compound of formula VII, is administered once, two or three times a day, for a total dose of 100 to 300 mg daily preferably of 220 to 230mg, most preferably 225 mg daily.
15. Use or method according to any one of claims 7 to 14, wherein the compound of formula VII, is administered three times a day, for a total dose of 220 to 230 mg, preferably 225 mg daily, and preferably a dose of 70 to 80 mg most preferably 75 mg per administration.
16. An article of manufacture comprising packaging material, and Λ/-[(9S,10R,11F?,13R)- 2,3,10,11 ,12,13-hexahydro-10-methoxy-9-methyl-1 -oxo-9, 13-epoxy-1 H,9H- diindolo[1 ,2,3-gh:3\2\1Mm]pyrrolo[3,4-j][1 ,7]benzodiazonin-11-yl]-N- methylbenzamide of the formula (VII) as defined in claim 8 or a pharmaceutically acceptable salts thereof, contained within said packaging material, wherein said packaging material comprises label directions which indicate that said compound of formula (VII), or said pharmaceutically-acceptable salt, is to be administered to mammals suffering from diseases involving SPTBN 1-FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product, in an amount from 100 to 300 mg, preferably 220 to 230mg, most preferably 225 mg following a specific dosage regimen to inhibit the development of diseases involving SPTBN1- FLT3 translocation-modulated signalling pathways and / or SPTBN 1-FLT3 translocation gene product.
17. An article of manufacture according to claim 16 wherein the compound of formula VII is administered three times a day, for a total dose of 220 to 230 mg preferably 225 mg daily, and preferably a dose of 70 to 80 mg most preferably 75 mg per administration for treating atypical chronic myeloid leukemia.
18. An isolated polynucleotide having a sequence encoding a SPTBN1-FLT3 translocation gene product.
19. A vector comprising a polynucleotide of claim 18.
20. A host cell comprising the vector of claim 18.
21. An organism containing a polynucleotide of claim 18.
22. The polynucleotide of claim 18, further comprising a polynucleotide sequence encoding a SPTBN 1-FLT3 fusion polypeptide
23. An isolated polypeptide having a sequence substantially identical to all or part of the polypeptide sequence of SEQ ID NO:2.
24. The use of a polynucleotide having a sequence encoding a SPTBN1-FLT3 fusion product as a drug target.
25. A method of detecting the presence of a SPTBN1-FLT3 translocation in a biological sample.
26. The method of claim 25 comprising generation and detection of a PCR product spanning the SPTBN 1-FLT3 translocation break point.
27. The method of claim 25, wherein FISH probes comprising genomic sequences capable of specifically hybridising to the FLT3 and SPTBN 1 genomic sequences are used to detect the presence of the SPTBN1-FLT3 translocation.
28. The method of claim 25 comprising detection of the SPTBN 1-FLT3 fusion gene product, wherein the gene product is selected from a transcribed SPTBN 1-FLT3 fusion mRNA and an encoded SPTBN 1-FLT3 fusion polypeptide.
29. The methods of claim 25-28 where the translocation is detected in a biological sample comprising at least one atypical chronic myeloid leukaemia cell.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019108492A1 (en) * 2017-11-30 2019-06-06 Albert Einstein College Of Medicine, Inc. Prevention and treatment of viral infections

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020086311A1 (en) * 1996-10-18 2002-07-04 Takara Shuzo Co., Ltd. Nucleic acid encoding receptor type protein kinase
WO2003037347A1 (en) * 2001-10-30 2003-05-08 Novartis Ag Staurosporine derivatives as inhibitors of flt3 receptor tyrosine kinase activity
WO2005014004A1 (en) * 2003-08-08 2005-02-17 Novartis Ag Combinations comprising staurosporines

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020086311A1 (en) * 1996-10-18 2002-07-04 Takara Shuzo Co., Ltd. Nucleic acid encoding receptor type protein kinase
WO2003037347A1 (en) * 2001-10-30 2003-05-08 Novartis Ag Staurosporine derivatives as inhibitors of flt3 receptor tyrosine kinase activity
WO2005014004A1 (en) * 2003-08-08 2005-02-17 Novartis Ag Combinations comprising staurosporines

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; November 2004 (2004-11), KERN WOLFGANG ET AL: "Identification of biologically distinct and clinically relevant subentities in patients with acute myeloid leukemia and normal karyotypes by use of gene expression profiling" XP002428153 Database accession no. PREV200510268168 & BLOOD, vol. 104, no. 11, Part 1, November 2004 (2004-11), page 60A, 46TH ANNUAL MEETING OF THE AMERICAN-SOCIETY-OF-HEMATOLOGY; SAN DIEGO, CA, USA; DECEMBER 04 -07, 2004 ISSN: 0006-4971 *
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; November 2005 (2005-11), GRAND FRANCIS H ET AL: "SPTBNI-FLT3 in atypical chronic myeloid leukemia transforms Ba/F3 cells to IL-3 independence and is sensitive to both tyrosine kinase inhibitors and immunotherapy" XP002428062 Database accession no. PREV200600184379 & BLOOD, vol. 106, no. 11, Part 1, November 2005 (2005-11), pages 568A-569A, 47TH ANNUAL MEETING OF THE AMERICAN-SOCIETY-OF-HEMATOLOGY; ATLANTA, GA, USA; DECEMBER 10 -13, 2005 ISSN: 0006-4971 *
STONE RICHARD M ET AL: "Patients with acute myeloid leukemia and an activating mutation in FLT3 respond to a small-molecule FLT3 tyrosine kinase inhibitor, PKC412." BLOOD 1 JAN 2005, vol. 105, no. 1, 1 January 2005 (2005-01-01), pages 54-60, XP002428058 ISSN: 0006-4971 *
WEISBERG ELLEN ET AL: "Inhibition of mutant FLT3 receptors in leukemia cells by the small molecule tyrosine kinase inhibitor PKC412." CANCER CELL JUN 2002, vol. 1, no. 5, June 2002 (2002-06), pages 433-443, XP002428059 ISSN: 1535-6108 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019108492A1 (en) * 2017-11-30 2019-06-06 Albert Einstein College Of Medicine, Inc. Prevention and treatment of viral infections
US11382919B2 (en) 2017-11-30 2022-07-12 Albert Einstein College Of Medicine Prevention and treatment of viral infections

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