EP3174541A1 - Dérivés de la thymine et de la chinazoline-dione pour inhiber hsp27 - Google Patents

Dérivés de la thymine et de la chinazoline-dione pour inhiber hsp27

Info

Publication number
EP3174541A1
EP3174541A1 EP15744197.3A EP15744197A EP3174541A1 EP 3174541 A1 EP3174541 A1 EP 3174541A1 EP 15744197 A EP15744197 A EP 15744197A EP 3174541 A1 EP3174541 A1 EP 3174541A1
Authority
EP
European Patent Office
Prior art keywords
general formula
hsp27
radical
vii
thymine
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP15744197.3A
Other languages
German (de)
English (en)
Inventor
Michael Prof. Dr. SCHROEDER
Yixin Zhang
Jörg-Christian HEINRICH
Joachim Haupt
Sainitin DONAKONDA
Petra Dr. Lennig
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Technische Universitaet Dresden
Original Assignee
Technische Universitaet Dresden
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Technische Universitaet Dresden filed Critical Technische Universitaet Dresden
Publication of EP3174541A1 publication Critical patent/EP3174541A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D279/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one sulfur atom as the only ring hetero atoms
    • C07D279/10—1,4-Thiazines; Hydrogenated 1,4-thiazines
    • C07D279/14—1,4-Thiazines; Hydrogenated 1,4-thiazines condensed with carbocyclic rings or ring systems
    • C07D279/18—[b, e]-condensed with two six-membered rings
    • C07D279/22—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/52—Purines, e.g. adenine
    • A61K31/522—Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
    • 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
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/5415—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame ortho- or peri-condensed with carbocyclic ring systems, e.g. phenothiazine, chlorpromazine, piroxicam
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7076—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid
    • A61K31/708—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid having oxo groups directly attached to the purine ring system, e.g. guanosine, guanylic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/52—Two oxygen atoms
    • C07D239/54—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals
    • C07D239/545—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/553—Two oxygen atoms as doubly bound oxygen atoms or as unsubstituted hydroxy radicals with other hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms with halogen atoms or nitro radicals directly attached to ring carbon atoms, e.g. fluorouracil
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
    • C07D239/72—Quinazolines; Hydrogenated quinazolines
    • C07D239/95—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in positions 2 and 4
    • C07D239/96—Two oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D279/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one sulfur atom as the only ring hetero atoms
    • C07D279/10—1,4-Thiazines; Hydrogenated 1,4-thiazines
    • C07D279/14—1,4-Thiazines; Hydrogenated 1,4-thiazines condensed with carbocyclic rings or ring systems
    • C07D279/18—[b, e]-condensed with two six-membered rings
    • C07D279/22—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom
    • C07D279/24—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom with hydrocarbon radicals, substituted by amino radicals, attached to the ring nitrogen atom
    • C07D279/26—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom with hydrocarbon radicals, substituted by amino radicals, attached to the ring nitrogen atom without other substituents attached to the ring system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D473/00—Heterocyclic compounds containing purine ring systems
    • C07D473/26—Heterocyclic compounds containing purine ring systems with an oxygen, sulphur, or nitrogen atom directly attached in position 2 or 6, but not in both
    • C07D473/28—Oxygen atom
    • C07D473/30—Oxygen atom attached in position 6, e.g. hypoxanthine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
    • C07F9/65616—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings containing the ring system having three or more than three double bonds between ring members or between ring members and non-ring members, e.g. purine or analogs
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • C07H19/073—Pyrimidine radicals with 2-deoxyribosyl as the saccharide radical

Definitions

  • the present invention relates to novel thymine derivatives and their use as drugs for the selective inhibition of the heat shock protein HSP27 (HSPB1).
  • HSP27 heat shock protein
  • Some types of cancer are known to be poorly responsive to treatment with cytotoxic drugs from the outset. But even in patients with actually treatable cancers, the therapy may fail after some time. One possible reason is resistance, which makes the tumor cells insensitive to cytostatics. Some underlying biological processes are known.
  • heat shock proteins are evolutionarily highly conserved proteins, which are formed by the cell under sublethal stress conditions. Heat shock proteins are chaperones whose natural task is to restore proteins that have been denatured by heat or chemicals to their proper state or to prevent the denaturation of proteins. Due to their mode of operation, heat shock proteins are overexpressed in a variety of cancer cells. In recent years, therefore, heat shock proteins have emerged as promising targets for cancer therapy.
  • HSP27 The heat shock protein HSP27 (HSPB1) is involved in a variety of cellular processes, such as apoptosis (programmed cell death), DNA repair and recombination. HSP27 is overexpressed in the cell in a wide range of cancers, including prostate cancer, colorectal cancer, liver and breast cancer, and influences the course of the disease. It was found that an increased expression of HSP27 with an increased resistance to cytostatics, such as gemcitabine (2 ', 2'-Difluordesoxycytidin) or bortezomib (Ve- Icade), accompanied by chemotherapy. Due to the negative influence of HSP27 on chemotherapy, HSP27 is considered a promising additional target for cancer therapy, especially for the suppression of chemoresistance.
  • cytostatics such as gemcitabine (2 ', 2'-Difluordesoxycytidin) or bortezomib (Ve- Icade
  • JP 2010 215669 A or US 2012/294846 A1 is based on the suppression of gene expression of HSP27 by the direct intervention in the translation or transcription by means of intracellular application of antisense oligonucleotides or double-stranded small interfering (si) RNA. Tie it the nucleotide-based inhibitors to the DNA or RNA and thereby specifically prevent the formation of HSP27 in the cell.
  • WO 2013 114339 A1 discloses a combination therapy for the treatment of cancer on the basis of the synergetic effect of a nucleotide inhibitor for inhibiting the expression of HSP27 in conjunction with an inhibitor against the protein EGFR (epidermal growth factor receptor), such as erlotinib, or antifolates like pemetrexed.
  • the nucleotide inhibitor is preferably an antisense oligonucleotide or a double-stranded siRNA.
  • nucleotide-based inhibitors are their excess of negative charges and the associated high polarity of the drug molecules, whereby the bioavailability is considerably minimized. Even more problematic is the enormous chemical instability of nucleotide-based inhibitors, in particular of RNA-based inhibitors.
  • short nucleotide-based inhibitors have the disadvantage of being potentially immunogenic, whereby they also act, for example, by secretion on non-transfected cells (eg macrophages), thereby inducing a strong and harmful to the patient immune response in the organism.
  • nucleotide-based inhibitors are low molecular weight, organic compounds, since these usually have a high stability in a biological system and at the same time a good bioavailability. Especially in the case of highly specific compounds, the "off-target effects" typical of other methods (for example due to the otherwise necessary transfection) do not occur.
  • DE 10 2008 035 299 A1 describes a method herein, after which the low molecular weight organic compound (E) -5- (2-Bromovinyl) -2 '-Deoxyuridine (BVDU) interacts directly with the protein HSP27 and thereby inhibits the function of the HSP27.
  • BVDU as a weak HSP27 inhibitor thus increases the sensitivity to cytostatic-induced apoptosis so that pancreatic cancer patients receive a survival advantage by oral administration of BVDU.
  • Clinical studies in late-stage pancreatic cancer patients have shown that BVDU is safe and effective at a dose of 500 mg / day. However, BVDU with an administered elevated dose of 760 mg / day was found to be detrimental to lightweight patients.
  • the above-described compound BVDU and its derivatives in DE 10 2008 035 299 A1 are the only known low molecular weight inhibitors of HSP27.
  • the active ingredients present have a considerable potential for improvement in terms of binding affinity.
  • WO 2009/156182 A2 describes uracil derivatives of the general formula I.
  • the object of the present invention is therefore to provide chemical compounds which inhibit the heat shock protein HSP27 as selectively and potently as possible, in particular for use in cancer therapy.
  • Purin derivatives according to the general formula (I) or (II) for use as medicaments, in particular for use for the treatment of carcinomas or cystic fibrosis, are specified for achieving the object:
  • R n is selected from hydrogen (H)
  • U is selected from oxygen (O), an optionally branched and / or OH-functionalized to C 4 -alkyl radical and optionally substituted vinyl radical, in particular substituted by halogen, particularly preferably F, Cl, I, preferably UO or is an optionally OH-functionalized d to C 4 -alkyl radical,
  • G is CH 2 , -CH 2 O- or O,
  • Y is O, S, NR, carboxyl, carbonyl or amide
  • R is an H or an optionally OH-functionalized C 1 to C 8 -alkyl radical
  • the substituent R y is H, OH, an optionally substituted to C 7 alkyl radical, an optionally substituted cyclic or polycyclic aryl radical or an optionally polycyclic nitrogen heterocycle, in particular an optionally substituted phenyl, naphthalene, bisphenyl, phenanthrene, benzopyrene, pyridine, diazine, triazole, piperidine, bipiperidine, piperazine, xanthene, carbazole, 9,10-dihydrophenanthrene, phenothiazines, triphenylmethane A radical or a combination thereof, wherein the substituents are selected from -F, -NH 2 , R and the group of the six-membered nitrogen heterocycle, in particular pyridine, 1, 2-diazine, 1, 3-diazine, piperidine, bipiperidine or Piperazine or a sequence of two to three in the para position to each other bridged above-mentioned substituents
  • the linker L is preferably used to restrict conformational flexibility (i.e.
  • Entropy reduction is selected from an optionally substituted phenyl, benzyl, pyridine radical, in particular substituted by -CH 3 , -CF 3 , halogen, especially preferably substituted by -CF 3 and F,
  • G is CH 2 , -CH 2 O- or O;
  • the linker L is selected from optionally substituted C1 to C6 -
  • Y is O, S, NR, carboxyl, carbonyl or amide
  • R is an H or an optionally substituted and / or branched C 1 to C 8 -alkyl radical
  • the substituent R y is as defined above or below H, OH, an optionally substituted cyclic or polycyclic aryl radical or an optionally substituted oxygen or nitrogen heterocycle, wherein the substituents are selected from -F, -NH 2, R and the group the six-membered nitrogen heterocycle, in particular pyridine, 1, 2-diazine, 1, 3-diazine, piperidine, bipiperidine or piperazine or a sequence of two to three above-mentioned substituents bridged in the para position relative to one another;
  • R a , R b and R c is H and at least one of the radicals is a cyclic radical which is preferably as selected above;
  • the substituent R N in formula (IV) is H or a benzoyl radical.
  • the substituent R y on the purine derivative of the formula (I) or (II) or the thymine derivative of the formula (VI), (VI '), (VII) or (VIII) taking into account the sterik of - adapted to the drug binding pocket, that the substituent R y represents a planar system.
  • the planar system does not necessarily have to be a conjugated system.
  • the substituent R y on the purine derivative of the formula (I) or (II) or the thymine derivative of the formula (VI), (VI '), (VII) or (VIII) is an optionally substituted cyclic or polycyclic aryl radical or a nitrogen heterocycle, in particular selected from among optionally substituted phenyl, naphthalene, bisphenyl, phenanthrene, benzopyrene, pyridine, diazine, xanthene, carbazole, phenothiazine, triphenylmethane, piperidine , Bipiperidine, piperazine residues and combinations thereof.
  • the substituents of the cyclic or polycyclic aryl radical or of the nitrogen heterocycle preferably have an -I and / or M effect, taking into account the sterics, as a result of which the purine derivatives of the formula (I) or (II) and the thymine -Derivat of formula (VI), (VI '), (VII) or (VIII) advantageously have a higher binding affinity for the drug binding pocket of the HSP27 protein.
  • the substituents of the cyclic or polycyclic aryl radical or the nitrogen heterocycle are selected from -halo, -NO 2 , -CN, -N (R) 2 , -SR, -OR, -COOR, -COR, R, where R is as above defined H or C1 to C8 alkyl.
  • the substituents of the cyclic or polycyclic aryl radical or of the nitrogen heterocycle are particularly preferably independently of one another substituents which have an -I and / or -M effect selected from H, -NO 2 , -CF 3 , -F, -Cl, - Br, -OH, -COOH, -OCH 3 and -COR, most preferably the substituents are independently selected from H, -NO 2 , -CF 3 , -OH, -COOH and F.
  • K is preferably selected independently of R y from the same radicals as mentioned above for R y . This is especially true when R y is H or OH.
  • the purine derivative of the formula (I) or (II) preferably has at least one optionally substituted cyclic or polycyclic radical aryl radical or an optionally substituted nitrogen heterocycle in R n or R y in addition to the purine structure.
  • the thymine derivative of the formula (VI), (VI '), (VII) or (VIII) preferably has, in addition to the thymine basic structure, at least one optionally substituted alliphatic cyclic or polycyclic radical (preferably in L) or an optionally substituted cyclic radical or polycyclic aryl radical or one or an optionally substituted oxygen or nitrogen heterocycle (preferably in K or R y ).
  • the nitrogen heterocycle is further substituted.
  • the nitrogen heterocycle is a triazole ring, which is preferably attached to Y in one position or to -CH in CH, and is preferably substituted in position 4 of the triazole ring.
  • Preferred substituents have the formula -R T -E with R T selected from a single bond and C 1 to C 5 alkyl and E selected from cyclic radicals.
  • the cyclic radicals in E are preferably monocyclic Radicals, particularly preferably selected from cyclopentyl, pyridines and phenyl and mono- or polysubstituted halogen (preferably Br, Cl or F) substituted phenyl.
  • the linker L is an optionally substituted phenyl, benzyl, pyridine radical, in particular substituted by -CH 3 , -CF 3 , halogen, particularly preferably substituted by -CF 3 and -F,
  • conformational flexibility i.e., entropy reduction
  • a "polycyclic aryl radical” is to be understood as meaning a conjugated system which contains at least two arylene groups which are directly linked to one another, ie in conjugation with one another. CH) are directly connected to one another and thus bring about the planarity of the system, such as, for example, xanthenes, carbazoles, phenothiazines or triphenylmethane.
  • purine derivatives according to the general formula (I) or (II) and thymine derivatives of the formula (VI), (VI '), (VII) or (VIII) inhibit the HSP27 protein by a specific interaction with the Drug binding pocket of the HSP27 protein.
  • the advantage of the purine derivatives according to the general formula (I) or (II) and the thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII) for use in cancer therapy is that these compounds have at least 50-fold higher activity for the selective inhibition of HSP27 than currently known compounds.
  • the purine derivatives of the general formula (I) or (II) and thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII) already have an activity in the lower one compared to HSP27 micromolar or submicromolar drug concentration range.
  • HSP2 means for the purposes of the present invention that the HSP27 by interaction with a low molecular weight compound, preferably the purine derivative of the general formula (I) or (II), the thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII) or phenothiazine derivatives according to the formula (V) loses the ability to interact with cancer-causing binding partners (for example proteins) and at the same time advantageously induces the initiation of apoptosis (for example by stimulated activity of Caspas).
  • cancer-causing binding partners for example proteins
  • Caspases are the most important enzymes of apoptosis in animals (programmed cell death).
  • the present invention is based on the recent finding that HSP27, which is overexpressed in many cancers and adversely affects the course of the disease, has a drug binding pocket for low molecular weight, organic compounds, which binds to the functionality (eg binding to client proteins and probably also the oligomerization) of the HSP27 protein and its activity is central.
  • the functionality of the drug binding pocket is based on two phenylalanine residues at positions 29 and 33 (Phe29 and Phe33) in the amino acid sequence of the protein.
  • the drug binding pocket of the HSP27 protein is a localized region (local tertiary structure) formed by the spatial arrangement of the amino acid sequence and having phenylalanine residues Phe29 and Phe33 (relative to the primary structure of HSP27).
  • the drug binding pocket of HSP27 is capable of interacting with a low molecular weight organic compound, preferably via coordinative interaction, such that the HSP27 protein loses its functionality (ability to oligomerize and interact with client proteins).
  • a client protein is a substrate, preferably a protein, on which HSP27 acts as a chaperone.
  • the binding affinity is a measure of the binding strength between binding partners (here the low molecular weight, organic compound) and a target protein.
  • the inhibitory concentration (K,) of the binding partner is the concentration at which complete inhibition of the target protein is present. It is understood that the lower K, the higher the binding affinity.
  • organic compounds can be pre-determined via a virtual screening, which interact much more specifically with the drug binding pocket of HSP27 than known compounds.
  • virtual screening encompasses a drug discovery process known to those skilled in the art in which new drug molecules are to be found by in silico methods (i.e., in computer experiments).
  • the binding affinity (Kj) of these potential inhibitors is not measured by wet-chemical experiments, as in a classical screening, but predicted by computer-based methods.
  • the structure of the HSP27 protein may undergo changes. Often these changes are limited to side-chain conformation, but whole groups of amino acids can move with their peptide backbone.
  • a few low molecular weight organic compounds must subsequently be experimentally probed by in vitro (ie, outside a living organism), in situ (ie, in cells, preferably in cell cultures), and / or in vivo (ie, in the living organism) be tested for their physicochemical affinity for the drug binding pocket of HSP27, which increases the efficiency for the detection of specific HSP27 inhibitors while reducing costs.
  • Such in vitro tests for determining the affinity towards the drug binding pocket of HSP27 are carried out, for example, via binding assays in which the association of two unlabelled binding partners are measured (eg, bio-layer interferometry).
  • the function of the target protein or its inhibition can also be quantified in aggregation assays.
  • the purine derivatives according to the invention of the general formula (I) or (II) and thymine derivatives of the formula (VI), (VI '), (VII) or (VIII) compared to HSP27 protein have a dissociation constant (K D ) in the range of 10 nM / L to 1000 ⁇ / L, preferably in the range of 100 nM / L to 800 ⁇ / L have.
  • the inhibitory effect of the pre-determined low molecular weight organic compounds on the functionality of the HSP27 protein is preferably measured in cells expressing HSP27.
  • the in situ methods are preferably carried out with the aid of conventional, readily cultivable cell lines in which the binding partners are expressed. Suitable cell lines are U937 and RPMI-8226 as examples of cancer cell lines and CCD-186Sk as an example of a cystic fibrosis cell line.
  • the detection of the activity of the low molecular weight, organic compounds is preferably carried out here by measuring the development of resistance to a cytostatic or else determining the interaction between HSP27 and its binding partners (for example via the measurement of caspase 3 activity).
  • low molecular weight organic compounds are understood as meaning molecular compounds which are composed primarily of the elements carbon, hydrogen, oxygen and nitrogen and preferably have a molecular weight in the range from 100 to 900 g / mol, in particular from 150 to 750 g / mol , especially 200 to 600 g / mol.
  • Low molecular weight organic compounds have the significant advantage over large molecules (e.g., nucleotide-based inhibitors such as antisense oligonucleotides or RNA oligonucleotides) of having much better handleability and stability under physiological conditions. Furthermore, low molecular weight, organic Compounds usually cell-like and therefore easier to apply in the cell or an organism.
  • the substituent R y is an optionally substituted cyclic or polycyclic aryl radical or an optionally polycyclic and / or substituted nitrogen heterocycle, which is in particular selected from a phenyl, naphthalene, bisphenyl, phenanthrene, benzopyrene, pyridine, Diazine, triazole, piperidine, bipiperidine, piperazine, xanthene, carbazole, phenothiazines, 9,10-Dihydrophenanthren-, triphenylmethyl radical or a combination thereof.
  • the six-membered nitrogen heterocycle is covalently bonded via a nitrogen atom to the general formula (I) or (II).
  • the substituent R y on the thymine derivative according to the general formula (VI), (VI '), (VII) or (VIII) is selected from a phenyl, triazole, piperazine , Diazine and triphenylmethyl radical, which may be substituted in particular with a six-membered nitrogen heterocycle, in particular pyridine, 1, 2-diazine, 1, 3-diazine, piperidine, bipiperidine or piperazine or a sequence of two to three in para position to each other bridged above-mentioned substituents.
  • the substituent R y on the thymine derivative according to the general formula (VI) or (VII) is H or OH, ie a sterically unbranched radical, where A 3 is particularly preferably -CHK-, ie a sterically demanding radical, wherein K is an optionally substituted five-membered nitrogen heterocycle, in particular triazole, diazole or imidazole, wherein the five-membered nitrogen heterocycle is preferably covalently bound to A 3 via a nitrogen atom.
  • the substituent on the five-membered nitrogen heterocycle is in particular selected from a phenyl, pyridine and diazine radical.
  • X is N or CH
  • R x is an optionally substituted and / or branched to C 4 -alkyl radical
  • R, R 2 , R 3 and R 4 independently of one another are -H, -halogen, -NO 2 , -CN, -NR 2 , and -SR, -OR, -COOR, -COR, -R or an optionally substituted one to C 4 vinyl radical or the optionally substituted aryl radical, where R is as defined above H or to C 8 alkyl radical.
  • R y is selected from -CR a R b R c wherein R a , Rb and R c are independently selected from H and cyclic radicals, preferably selected from optionally substituted cyclic or polycyclic aryl radicals and optionally substituted heterocycles, more preferably H, Phenyl, napthylene, biphenyl. At least one of the radicals R a , R b and R c is preferably H and at least one of the radicals is a cyclic radical which is preferably selected as above.
  • the purine derivatives according to the general formula (I) and (II) or thymine derivatives of the formula (VI), (VI '), (VII) and (VIII) particularly advantageous activity in the lower micromolar or submicromolar, more preferably in the nanomolar concentration range.
  • the substituents R 1 to R 4 taking into account the steric an -I and / or -M effect, whereby the purine derivatives according to the general formula (I) or (II) or Thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII) advantageously have a high binding affinity for the drug binding pocket of the HSP27 protein (ie low K, and K D ).
  • the substituents R 1 to R 4 are preferably, independently of one another, substituents which have an -I and / or -M- Having effect selected from H, -NO 2 , -CF 3 , -F, -Cl, -Br, -OH, -COOH, -OCH 3 and -COR, particularly preferably the substituents are independently selected from H, - N0 2 , -CF 3 , -OH, -COOH and F. According to a very particularly preferred embodiment of the present invention, only two, more preferably only one of the substituents R 1 to R 4 have a different substituent from H.
  • one of the substituents R 1 to R 4 is an optionally substituted aryl radical which has a pronounced + M effect, preferably a phenyl radical or a six-membered nitrogen heterocycle, selected from pyridine, pyridazine, pyrimidine or pyrazine.
  • one of the substituents R 1 to R 4 is an optionally substituted six-membered nitrogen heterocycle.
  • R is preferably n a sterically less demanding, less substituent selected from H, an optionally branched C r C 4 alkyl or ether radical, wherein VH or OH, an optionally substituted five- or six-membered ring-oxygen Hetrozyklus.
  • such purine derivatives of the general formula (I) and (II) wherein R n is as defined above have a K, in the range of 10 nmol / L to 5 mmol / L, more preferably in the range of 50, compared to HSP27 nmol / L to 5 mmol / L. It may alternatively be provided that the substituent R n is essentially made up of two parts, the part 1 U and W and the part 2 A comprises.
  • the part is 1 (U and W) is an optionally branched C r C 4 alkyl or ether radical
  • the portion 2 (V) is a substituted six-membered nitrogen heterocycle such as pyridine, diazine, piperidine or piperazine
  • such purine derivatives of the general formula (I) and (II) in which R n is as defined above have a Kj in the at least the lower micromolar or submicromolar range compared to HSP27. rich, more preferably in the nanomolar range, most preferably in the range of 10 nmol / L to 200 nmol / L on.
  • V is an amino acid AS which is covalently bonded to W via its carboxyl group.
  • R 1 , R 3 and optionally R 4 of the substituent R y to purine derivatives according to the general formula (I) or (II) or thymine derivatives according to the general formula (VI) H are particularly preferred.
  • variations of purine derivatives according to the general formula (I), (II) and thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII) showed particularly high binding affinity for the drug-binding pocket of the HSP27 protein in which the phenyl ring according to formula (IV) has only the substituent R 2 in the meia position and where R 1 and R 3 are H.
  • R 1 and R 3 H and R 2 is an optionally substituted phenyl radical.
  • purine derivatives according to the general formula (I) and (II) or thymine derivatives according to the general formula (VI), (VI '), (VII) and (VIII) have only the substituent R in orffto position on the phenyl ring according to the formula (IV) and / or R 3 in the para position, where R 2 in the mefa position is H.
  • R 1 and R 2 are H and R 3 is an optionally substituted phenyl radical.
  • R 2 , R 2 and R 3 are independently of each other R or an optionally substituted C 4 to C 4 vinyl radical, where R is H or an optionally OH- functionalized C 1 -C 5 -alkyl radical.
  • R 1 , R 3 and R 4 are H and R 2 is H, -halo, -COR or an optionally substituted phenyl radical.
  • HP27 inhibitors in particular selected from the group of purine derivatives of the general formula (I) or (II), the thymine derivatives according to the general formula (VI) and / or phenothiazine derivatives of the general formula (V) for the suppression of chemoresistance formation in cancer therapy and for use in the treatment of cystic fibrosis.
  • Cystic fibrosis is a hereditary disease in which, in most cases of cystic fibrosis, the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) at position 508, due to a deletion of three nucleotides in the CFTR-encoding DNA sequence, is the amino acid phenylalanine (Phe508). is missing.
  • the CFTR protein is a transmembrane protein that regulates water and salt transport in the plasma membrane of epithelial cells.
  • the resulting protein of the deletion mutant AF508 is not folded correctly during its formation (translation). Therefore, it is not transported via the endoplasmic reticulum to the cell membrane, but encapsulated with the involvement of HSP27 and fed to the degradation.
  • the purine derivatives of the general formula (I) and (II) or thymine derivatives according to the general formula (VI), (VI '), (VII) and (VIII) are suitable.
  • phenothiazine derivatives of the general formula (V) as medicaments for use in cancer therapy, in particular for the suppression of chemoresistance formation, and in the treatment of cystic fibrosis.
  • the invention also provides the use of these compounds for the preparation of a medicament, preferably for use in cancer therapy, in particular for the suppression of chemoresistance formation, and in the treatment of cystic fibrosis.
  • Also of the present invention are compounds such as pharmacologically acceptable salts, prodrugs, enantiomers, diastereomers, racemic mixtures, crystalline forms, amorphous forms and solvates, comprising a purine derivative according to the general formula (I) and (II) or a thymine A derivative of general formula (VI), (VI 1 ), (VII) and (VIII), for use as a medicament in cancer therapy, in particular for suppressing chemoresistance formation, and the treatment of cystic fibrosis.
  • a pharmacologically acceptable salt includes chemical compounds of the purine derivative according to the general formula (I) and (II) or of the thymine derivative of the formula (VI), (VI '), (VII) and ( VIII) from positively and negatively charged ions, which can be obtained, for example, by treatment with a weak, pharmaceutically acceptable acid or base, depending on the substituents of the purine derivative.
  • a prodrug is an inactive or poorly active pharmacological substance which is first converted by a chemical modification under physiological conditions (ie by metabolism or metabolization) in the organism into the pharmacologically active form of a purine derivative according to the general formula (I) or (II ) or a thymine derivative according to the general formula (VI), (VI '), (VII) and (VIII) is converted. It can also be provided that the prodrug by chemical or biochemical methods in an ex vivo environment into the pharmacologically active form of a purine derivative according to the general formula (I), (II) or a thymine derivative according to the general formula (VI), (VI 4 ), (VII) or (VIII) is transferred.
  • a particularly advantageous property of the purine derivatives according to the general formula (I) or (II) and the thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII) compared to conventional nucleotide based inhibitors is that they have as low molecular weight, organic compounds in the context of the invention, for example, a largely independent of the gastric pH bioavailability.
  • compounds containing a purine derivative according to the general formula (I) and / or (II) and / or a thymine derivative of the formula (VI) or (VII), an organism by administered orally or parenterally and resorbed via the mucous membranes are suitable, for example, in the form of a tablet, capsule or liquid or rectally in the form of suppositories.
  • Examples of organisms for the purposes of the present invention are preferably selected from the group of mammals, wherein the purine derivative according to the general formula (I) or (II) and / or thymine derivative of the formula (VI), (VI ') , (VII) or (VIII) can very particularly preferably be applied to a human.
  • Another object of the invention relates to a pharmaceutical formulation containing at least one purine derivative according to the general formula (I) or (II) and / or thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII), for use in cancer therapy or the treatment of cystic fibrosis.
  • the purine derivatives according to the general formula (I) or (II) and / or the thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII) in the pharmaceutical formulation be used as the only pharmacologically active ingredient or in combination with at least one cytostatic, such as to broaden the spectrum of action or to prevent development of resistance. In many cases this results in additive or synergistic effects, i. the effectiveness of the mixture is greater than the effectiveness of the individual components.
  • the pharmaceutical formulation according to the invention only an active ingredient in the form of the purine derivative according to the general formula (I) or (II) or the thymine derivative of the general formula (VI) or (VII).
  • a pharmaceutical formulation according to the invention comprising the purine derivative according to the general formula (I) or (II) and / or the thymine derivative of the general formula (VI), (VI '), (VII) or (VIII) in combination with at least one cytostatic.
  • a pharmaceutical formulation according to the invention thus offers the particular advantage that the formation of resistance in the case of a cytostatic treatment is prevented or at least significantly delayed.
  • cytostatic agents are: folic acid antagonists (eg methotrexate, pemetrexed), pyrimidine analogs (for example 5-fluorouracil, gemcitabine), purine analogs (for example pentostatin, azathioprine) and N- or C-terminally protected oligopeptides (for example bortezomib).
  • folic acid antagonists eg methotrexate, pemetrexed
  • pyrimidine analogs for example 5-fluorouracil, gemcitabine
  • purine analogs for example pentostatin, azathioprine
  • N- or C-terminally protected oligopeptides for example bortezomib
  • the purine derivative of the general formula (I) or (II) and / or the thymine derivative of the general formula (VI), (VI '), (VII) or (VIII) can be administered orally in the form of tablets, capsules, liquids or syrups or rectally in the form of suppositories.
  • R 5 and R 6 independently of one another are H, an optionally substituted piperazine, phenyl (-C 6 H 5 ), hydroxyphenyl (-OC 6 H 5 ), phenylene radical (-C 6 H 4 -),
  • phenothiazine derivatives are also able to selectively bind to the drug binding pocket of the HSP27 protein and thereby inhibit the functionality of the HSP27 protein so that phenothiazine derivatives are suitable for use in cancer therapy or the treatment of cystic fibrosis.
  • Phenothiazine derivatives according to the general formula (V) are used in pharmacy in a known manner as antipsychotics, preferably in a concentration of 1, 0 to 3.0 mg per kg body weight of an organism.
  • the combination of a purine derivative according to the general formula (I) or (II) and / or a thymine derivative of the general formula (VI), (VP), (VII) or (VIII) and a phenothiazine derivative according to the general formula (V) in the selective inhibition of the HSP27 protein advantageously lead to superadditive ("synergistic") effects, for example the following effects are possible: lent beyond the actual expected effects: reduced use concentration and / or extended spectrum of activity and / or increased efficacy of the individual compounds (ie active ingredients) and pharmaceutical formulations.
  • the pharmaceutical formulation according to the invention therefore comprises the purine derivative according to the general formula (I) or (II) and / or the thymine derivative of the general formula (VI), (VI '), (VII ) or (VIII) in combination with one or more phenothiazine derivatives according to the general formula (V).
  • Chlorpromazine showed only a low affinity for interaction with HSP27 in wet-chemical experiments and is therefore less preferred as a representative of the phenothiazine derivatives of the general formula (V) for use according to the invention in cancer therapy or the treatment of cystic fibrosis.
  • phenothiazine derivatives of the general formula (V) are used singly or as a mixture thereof as medicaments for use in cancer therapy or the treatment of cystic fibrosis.
  • the invention therefore also relates to the use of the phenothiazine derivatives of the general formula (V) or a mixture thereof for the preparation of a pharmaceutical formulation for use in cancer therapy or the treatment of cystic fibrosis.
  • the phenothiazine derivatives of general formula (V) are administered in an amount of 0.1 to 50 mg / kg, preferably about 0.1 to 30 mg / kg of body weight per day, and ideally in an amount of 0.5 to 20 mg / kg, more preferably 0.5 to 15 mg / kg or 0.3 ⁇ / kg to 150 mol / kg and ideally 1, 5 pmol / kg to 60 pmol / kg of body weight per day.
  • the correct dosage can be determined both by examining the efficacy of the compound in the cell proliferation assays and by determining the toxicity in animal (and ultimately human) studies.
  • the present invention further encompasses a method for reducing the functionality of the HSP27 protein in a cell or an organism, comprising administering a purine derivative according to the general formula (I) or (II) or a thymine derivative of the general formula (VI), (VI '), (VII) or (VIII) in a physiological effective concentration for inhibiting the HSP27 protein, wherein the purine derivative interacts with the drug binding pocket of the HSP27 protein.
  • the invention also includes the use of a purine derivative according to the general formula (I), (II) or a thymine derivative of the general formula (VI), (VI '), (VII) or (VIII) and / or Phenothiazine derivatives of general formula (V) for the treatment of conditions associated with increased HSP27 signaling, particularly in cancer as well as cystic fibrosis.
  • the invention also includes a method for the treatment of cystic fibrosis or prevention of chemoresistance formation in cancer therapy by administration of an effective dose of an HSP27 inhibitor, in particular a purine derivative according to the general formula (I), (II) or a thymine derivative of the general formula (VI) or (VII) (VI), (VI '), (VII) or (VIII) and / or phenothiazine derivatives of the general formula (V).
  • an HSP27 inhibitor in particular a purine derivative according to the general formula (I), (II) or a thymine derivative of the general formula (VI) or (VII) (VI), (VI '), (VII) or (VIII) and / or phenothiazine derivatives of the general formula (V).
  • the method of treatment comprises the administration of a pharmaceutical formulation containing at least one purine derivative according to the general formula (I), (II), a thymine derivative of the formula (VI), (VI '), (VII) or (VIII) and / or a phenothiazine derivative of the general formula (V) in a physiologically effective concentration.
  • the pharmaceutical formulation for the application is solid or liquid, for example. In the form of a tablet, capsule or liquid or rectally in the form of suppositories such that the pharmaceutical formulation is administered to an organism by oral or parenteral administration and absorbed via the mucous membranes.
  • the purine derivatives according to the general formula (I), (II), thymine derivatives of the formula (VI), (VI '), (VII) or (VIII) and / or a phenothiazine derivatives of the general formula (V) in an amount of about 0.1 to 30 mg / kg of body weight per day, and ideally in an amount of 0.5 to 15 mg / kg of body weight per day.
  • mammals such as humans, into consideration.
  • the invention also purine derivatives according to the general formula (I) and / or (II), a thymine derivative of the formula (VI), (VI '), (VII) or (VIII) and / or a phenothiazine Derivatives of the general formula (V) or of a mixture for use as medicaments, in particular for use in the treatment of cancer or the treatment of cystic fibrosis.
  • the dosage of the phenothiazine derivatives of the general formula (V) and / or the purine derivatives of the general formula (I) or (II) and / or the thymine derivatives of the formula (VI), (VI '), (VII ) or (VIII) depends on various factors, for example the method of administration, age, weight and health, including the type of organism to be treated.
  • the compounds having as low a K as possible are particularly preferred. Preference is given to the compounds having a K of not more than 100 ⁇ / L, in particular not more than 10 ⁇ / L, particularly preferably not more than 500 nmol / L.
  • thymine derivatives according to the general formula (VI), (VI '), (VII) or (VIII): (with the respective calculated inhibitory concentration based on HSP27-K,):
  • the compounds with the least possible amount are particularly preferred. Preference is given to the compounds having a K of not more than 100 ⁇ / L, in particular not more than 10 pmol / L, particularly preferably not more than 500 nmol / L.
  • the present invention also encompasses the use of a purine derivative according to the general formula (I) and / or (II), a thymine derivative of the formula (VI), (VI '), (VII) or (VIII) and / or a phenothiazine derivative of the general formula (V) for the preparation of a pharmaceutical formulation, in particular for use in the treatment of cancer or the treatment of cystic fibrosis.
  • the invention also provides the use of a purine derivative according to the general formula (I), (II), a thymine derivative of the formula (VI), (VI '), (VII) or (VIII) and / or a Phenothiazine derivative of the general formula (V) in combination with chemotherapy, radiation therapy and / or cancer immunotherapy for use in the treatment of cancers, wherein advantageously reduces the resistance (ie resistance of a tumor or tumor cells compared to the applied treatment method) to the treatment methods , is suppressed in particular.
  • Cancer immunotherapy methods differentiate between active and passive immunization.
  • active immunization the patient is given substances that are supposed to trigger an immune response in his immune system.
  • Passive immunization uses antibodies or antibody fragments.
  • leukocytes are removed from the patient, cultured ex vivo, and then injected back into the patient.
  • the further treatment of cancers is made much more difficult by resistance formation.
  • a purine derivative according to the general formula (I), (II), a thymine derivative of the formula (VI), (VI '), (VII) or (VIII) and / or a phenothiazine derivative of the general formula ( V) thus offers the particular advantage that resistance formation in chemotherapy, radiation therapy or cancer immunotherapy, in particular cytostatic treatment, is prevented or at least significantly delayed.
  • cytostatic agents are: folic acid antagonists (eg methotrexate, pemetrexed), pyrimidine analogs (for example 5-fluorouracil, gemcitabine), purine analogs (for example pentostatin, azathioprine) and N- or C-terminally protected oligopeptides (for example bortezomib).
  • the purine derivative, thymine derivative and / or phenothiazine derivative is used prior to the initiation of chemotherapy, radiotherapy and / or cancer immunotherapy and the administration of the purine derivative, thymine derivative and / or phenothiazine derivative during the said cytotoxic therapies (chemotherapy, radiotherapy and / or cancer immunotherapy) continued to suppress the formation of resistance.
  • the administration of the purine derivative, thymine derivative and / or phenothiazine derivative is preferably started 15 minutes to 4 hours before the beginning of the cytotoxic therapy.
  • Fig. 1 a spectrophotometric protein aggregation assay at 43 ° C, a wavelength of 500 nm and a concentration of proteins: 1, 44 ⁇ / ⁇ _ citrate synthase (CS), 481 nmol / L HSP27 (HSP) in a 40th mmol / L Hepes buffer (pH 7.4) in the treatment with 14 or 28 mol / L 2- (3-trichlorovinyl-phenylamino) -1, 9-dihydro-purin-6-ones (CS + HSP + test substance No. 2), with 750 ⁇ / L BVDU (CS + HSP + BVDU) and without (CS + HSP; reference);
  • FIG. 2 shows the cell numbers over the number of days of treatment of U937 cells treated with the cytostatic bortezomib (0.1 to 0.5 ng / ml), in each case in combination with the HSP27 inhibitors acepromazine (+1 ⁇ M ACE) and BVDU (+30 BVDU) and without HSP27 inhibitor.
  • FIG. 3 the cell numbers over the number of days of treatment of U937 cells treated with the cytostatic bortezomib (0.2 to 0.4 ng / ml), in each case in combination with the HSP27 inhibitors acepromazine (+1 ⁇ ACE) and BVDU (+30 ⁇ BVDU) and without HSP27 inhibitor.
  • Fig. 4 shows the inhibition of resistance formation of the tumor cell line RPMI-8226 by acepromazine.
  • Figure 4a shows RPMI 8226 cells treated with cytostatic bortezomib 0.2-0.4 ng / ml (A), combination treatment with bortezomib plus 0.75 ⁇ acepromazine (B) and combination treatment with bortezomib plus 30 ⁇ BVDU (C).
  • Fig. 4b shows the control without cytostatic: RPMI 8226 cells untreated (A), 0.75 ⁇ acepromazine alone (B) and 30 ⁇ BVDU alone (C).
  • FIG. 5 shows the inhibition of resistance formation of the tumor cell line RPMI-8226 by chlorpromazine.
  • Figure 5a shows RPMI 8226 cells treated with cytostatic bortezomib 0.2-0.4 ng / ml (A), combination treatment with bortezomib plus 0.5 ⁇ chlorpromazine (B) and combination treatment with bortezomib plus 30 ⁇ BVDU (C).
  • Fig. 5b shows the control without cytostatic: RPMI 8226 cells untreated (A), 0.5 ⁇ chlorpromazine alone (B) and 30 ⁇ BVDU alone (C).
  • Figure 6 shows the inhibition of resistance formation of the tumor cell line RPMI-8226 by 2- (4-butylphenylamino) -9- (2-hydroxyethoxymethyl) -1,9-dihydro-purin-6-one.
  • 6a shows RPMI 8226 cells treated with cytostatic bortezomib 0.1-0.3 ng / ml (A), combination treatment with bortezomib plus 1 ⁇ M 2- (4-butylphenylamino) -9- (2-hydroxy-ethoxymethyl ) -1, 9-dihydro-purin-6-one (B) and combination treatment with bortezomib plus 30 ⁇ BVDU (C).
  • 6b shows the control without cytostatic: RPMI 8226 cells untreated (A), 1 ⁇ 2- (4-butylphenylamino) -9- (2-hydroxy-ethoxymethyl) -1,9-dihydro-purin-6 one alone (B) and 30 ⁇ BVDU alone (C).
  • FIG. 7 shows the inhibition of resistance formation of the tumor cell line RPMI-8226 by 2- (3-trichlorovinyl-phenylamino) -1, 9-dihydro-purin-6-ones.
  • FIG. 7a shows RPMI 8226 cells treated with cytostatic bortezomib 0.1-0.3 ng / ml (A), combination treatment with bortezomib plus 1 ⁇ M 2- (3-trichlorovinylphenylamino) -1,9-dihydro-purine 6-one (B) and combination treatment with bortezomib plus 30 ⁇ BVDU (C).
  • A cytostatic bortezomib 0.1-0.3 ng / ml
  • B 2- (3-trichlorovinylphenylamino) -1,9-dihydro-purine 6-one
  • C combination treatment with bortezomib plus 30 ⁇ BVDU
  • 7b shows the control without cytostatic: RPMI 8226 cells untreated (A), 1 ⁇ 2- (3-trichlorovinyl-phenylamino) -1, 9-dihydro-purin-6-one alone (B) and 30 ⁇ BVDU alone (C).
  • FIG. 8 shows the inhibition of the resistance formation of the tumor cell line RPMI-8226 by 9H-xanthene-9-carboxylic acid [4- (5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl). but-2-enyl] amide.
  • Fig. 8 shows the inhibition of the resistance formation of the tumor cell line RPMI-8226 by 9H-xanthene-9-carboxylic acid [4- (5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl). but-2-enyl] amide.
  • FIG. 8a shows RPMI 8226 cells treated with cytostatic bortezomib 0.075-0.275 ng / ml (A), combination treatment with bortezomib plus 1 ⁇ M 9H-xanthene-9-carboxylic acid [4- (5-methyl-2,4-dioxo-3 , 4-dihydro-2H-pyrimidin-1-yl) -but-2-enyl] -amide (B) and combination treatment with bortezomib plus 30 ⁇ M BVDU (C).
  • A cytostatic bortezomib 0.075-0.275 ng / ml
  • B 4-dihydro-2H-pyrimidin-1-yl
  • C combination treatment with bortezomib plus 30 ⁇ M BVDU
  • FIG. 8b shows the control without cyto- Assay: RPMI 8226 cells untreated (A), 1 ⁇ M 9H-Xanthene-9-carboxylic acid [4- (5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl) -but -2-enyl] -amide alone (B) and 30 ⁇ BVDU alone (C).
  • FIG. 9 shows the inhibition of the resistance formation of the tumor cell line RPMI-8226 by 2-biphenyl-4-yl-N- [4- (5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidine-1 - yl) -but-2-enyl] -acetamide.
  • 9a shows RPMI 8226 cells treated with cytostatic bortezomib 0.075-0.275 ng / ml (A), combination treatment with bortezomib plus 1 ⁇ 2-biphenyl-4-yl-N- [4- (5-methyl-2,4-) dioxo-3,4-dihydro-2H-pyrimidin-1-yl) -but-2-enyl] -acetamide (B) and combination treatment with bortezomib plus 30 ⁇ M BVDU (C).
  • 9b shows the control without cytostatic: RPMI 8226 cells untreated (A), 1 ⁇ M 2-biphenyl-4-yl-N- [4- (5-methyl-2,4-dioxo-3,4-dihydroxy) 2H-pyrimidin-1-yl) -but-2-enyl] -acetamide alone (B) and 30 ⁇ BVDU alone (C).
  • FIG. 10 shows the inhibition of the resistance formation of the tumor cell line U-937 by acepromazine.
  • 10a shows U-937 cells treated with cytostatic bortezomib 0, 1-0.5 ng / ml (A), combination treatment with bortezomib plus 0.75 ⁇ acepromazine (B) and combined treatment with bortezomib plus 30 ⁇ BVDU (C)
  • Fig. 11b shows the control without cytostatic: RPMI 8226 cells untreated (A), 0.75 ⁇ acepromazine alone (B) and 30 ⁇ BVDU alone (C).
  • FIG. 11 shows the inhibition of the resistance formation of the tumor cell line U-937 by 2- (4-butyl-phenylamino) -9- (2-hydroxy-ethoxymethyl) -1,9-dihydro-purin-6-one.
  • 1 1 a shows U-937 cells treated with cytostatic bortezomib 0.1-0.5 ng / ml (A), combination treatment with bortezomib plus 0.75 ⁇ 2- (4-butylphenylamino) -9- ( 2-hydroxy-ethoxymethyl) -1,9-dihydro-purin-6-one (B) and combination treatment with bortezomib plus 30 ⁇ M BVDU (C).
  • 11b shows the control without cytostatic: RPMI 8226 cells untreated (A), 0.75 ⁇ M 2- (4-butylphenylamino) -9- (2-hydroxy-ethoxymethyl) -1,9-dihydro-purine -6-one alone (B) and 30 ⁇ BVDU alone (C).
  • the method for detecting molecular bonds using gel filtration was described as "SpeedScreen” in 2004 [Muckenschnabel et al., 2004] and adapted according to the requirements of the invention
  • the separation method is that larger molecules, such as proteins, form a matrix of Sephadex G-25 happen while low-molecular-weight compounds (small molecules) penetrate into the matrix and do not pass through it.
  • PD SpinTraps G-25 from GE Healthcare was used. These columns allow molecules of molecular weight greater than or equal to 5000 g / mole to pass through the 2-minute centrifugation at 800 xg (as directed), while retaining smaller molecules in the matrix.
  • the target protein HSP27 has a molecular weight of 27000 g / mol and thus passes through the matrix.
  • the HSP27 inhibitors of the general formula (I) described according to the invention are low molecular weight compounds which can only pass through the Sephadex G-25 matrix if they have previously bound to the target protein HSP27 and are transported by it through the matrix. Unbound low molecular weight compounds are retained by the matrix. Molecules that have passed through the matrix can then be detected by mass spectroscopy. This allows the distinction between "binders" and "non-binders”.
  • acepromacin (as acepromacin maleate) was tested as a representative of phenothiazine derivatives using SpeedScreen positive.
  • the method for detecting molecular bonds using gel filtration was described as "SpeedScreen” in 2004 [Muckenschnabel et al., 2004] and adapted according to the requirements of the invention in that larger molecules, such as proteins, can pass through a matrix of Sephadex G-25, while low-molecular-weight compounds (small molecules) penetrate into the matrix and do not pass through it.
  • PD SpinTraps G-25 from GE Healthcare was used. These columns allow molecules of molecular weight greater than or equal to 5000 g / mole to pass through the 2-minute centrifugation at 800 xg (as directed), while retaining smaller molecules in the matrix.
  • the target protein HSP27 has a molecular weight of 27,000 g / mole and thus passes through the matrix.
  • the HSP27 inhibitors of the general formula (I) described according to the invention are low molecular weight compounds which can only pass through the Sephadex G-25 matrix if they have previously bound to the target protein HSP27 and are transported by it through the matrix. Unbound low molecular weight compounds are retained by the matrix. Molecules that have passed through the matrix can then be detected by mass spectroscopy. This allows the distinction between "binders" and "non-binders”.
  • Both thymine derivatives pass through the Sephadex G-25 matrix and are detected after passing through the medium.
  • Bio-Layer Interferometry is a technique for detecting biomolecular interactions. It is an analytical technique that measures the optical interference of white light reflected from two surfaces: a surface consists of a layer of the immobilized target protein on the tip of the biosensor and an internal reference surface. The binding of ligands to the target protein alters the interference pattern and can be measured in real time. A ForteBio Octet Red 384 instrument was used for the measurements. The target protein HSP27 was biotinylated and bound to the corresponding sensors (SSA). A big advantage of this analysis method is that the analytes do not have to be immobilized or labeled. The purine derivatives of the general formula (I) or (II) pre-determined by means of in silico screening are used for these measurements in a concentration range (K D ) between 333 / mol / L and 10 nmol / L.
  • K D concentration range
  • the citrate syntase denatures. This heat denaturation is prevented or delayed by the presence of HSP27 [Jakob U, 1993].
  • FIG. 1 shows that the protein citrate synthase (graph CS alone) is denatured by increasing the temperature to 43 ° C. and forms aggregates. The aggregation is measured in the spectrometer at 500 nm (light scattering by protein aggregates). The presence of HSP27 (HSP) counteracts aggregation (see Graph, CS + HSP). Since the addition of the test substances (BVDU and 2- (3-trichlorovinyl-phenylamino) -1, 9-dihydro-purine 6-on) inhibit the chaperone function of HSP27, the activity of the respective test substance can be determined by measuring the aggregation behavior of the CS.
  • HSP27 HSP27
  • the inhibition of HSP27 correlates with the binding strength of a low molecular weight organic compound to the drug binding pocket of HSP27.
  • the active concentration of a HSP27-interacting low molecular weight organic compound can be estimated by the activity of BVDU in BVDU equivalents, i. equal to HSP27 inhibiting dose in mg BVDU, to be converted.
  • the test substance 2- (3-trichlorovinyl-phenylamino) -1,9-dihydro-purin-6-one has a BVDU equivalent dose of less than 1.9 mg.
  • the heat-denatured protein aggregates of the citrate synthase from Example 1 are not water-soluble, they can be completely separated from the supernatant by centrifugation at room temperature at 16,000 ⁇ g, 10 min. Using capillary electrophoresis, the compositions of the separated precipitates can be visualized and quantified. The relative amount of protein precipitated citrate synthase serves as a measure of the effectiveness of the test substances.
  • Table 1 below shows the capillary electrophoretic separation of heat-denatured citrate synthase protein aggregates after thermal treatment of the samples at 43 ° C. for 120 minutes. rel. Protein amount effectiveness in
  • the four test substances proved to be significantly more effective at the use concentration of 10 ⁇ / L than the reference substance BVDU and can consequently be dosed in the range between 53 to 79 times lower in order to achieve a comparable effect compared to the conventional BVDU.
  • the purine derivative 2- (4-butylphenylamino) -9- (2-hydroxy-ethoxymethyl) -1,9-dihydro-purin-6-one shows a 69-fold higher activity for inhibiting HSP27 protein than BVDU (BVDU equivalent dose is 1, 4 mg).
  • the purine derivative 2- (3-trichlorovinyl-phenylamino) -1,9-dihydro-purin-6-one also shows a 53-fold higher activity for BVDU to inhibit the HSP27 protein, giving a BVDU equivalent dose of less Corresponds to 1, 9 mg.
  • HSP27 favors the development of chemoresistance by e.g. interacts with apoptosis proteins and thus prevents the targeted cell death of cancer cells.
  • bortezomib a modern cytostatic agent (proteasome inhibitor)
  • HSP27-driven resistance is well documented [Chauhan D, 2004].
  • U937 cells (histiocytic lymphoma) were cultured in DMEM culture medium (+ 10% FCS) at 37 ° C, 5% C0 2 in H 2 0-saturated atmosphere. 100,000 cells / ml were sown and incubated with the cytostatic bortezomib (0.1 ng / ml) and the respective test substance (dosage in the current experiment: 1 ⁇ / L acepromazine). The reference substance, BVDU was used at a concentration of 30 ⁇ / L. The cells were passaged each time before the cell count reaches 1,000,000 cells / ml.
  • Example 6 The experiments of Example 6 were repeated accordingly, wherein 11937 cells (histiocytic lymphoma) are cultured in DMEM culture medium (+ 10% FCS) at 37 ° C, 5% C0 2 in H 2 0 saturated atmosphere. 100,000 cells / ml were seeded and incubated with an initial concentration of 0.2 ng / ml of the cytostatic bortezomib and each without inhibitor and with acepromazine as HSP27 inhibitor at a concentration of 1 ⁇ / L. The reference substance, BVDU was used at a concentration of 30 ⁇ / L. The cells were passaged each time before the cell count reaches 1,000,000 cells / ml.
  • Example 8 Capillary electrophoresis of CS protein precipitates with Thymi n derivatives
  • the heat-denatured protein aggregates of citrate synthase from Example 2 are not water-soluble, they can be completely separated from the supernatant by centrifugation at room temperature at 16,000 ⁇ g, 10 min. Using capillary electrophoresis, the compositions of the separated precipitates can be visualized and quantified. The relative amount of protein precipitated citrate synthase serves as a measure of the effectiveness of the test substances.
  • Table 2 below shows the capillary electrophoretic separation of heat-denatured citrate synthase protein aggregates after thermal treatment of the samples at 43 ° C for 120 minutes.
  • the four test substances proved to be significantly more effective than the reference substance BVDU at the use concentration of 10 ⁇ mol / L each and can consequently be dosed in the range from 49 to 79 times lower in order to achieve a comparable effect compared to the conventional BVDU.
  • the thymine derivative 2-biphenyl-4-yl-N- [4- (5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl) -but-2-enyl] acetamide shows a 61-fold higher activity to inhibit the HSP27 protein than BVDU (BVDU equivalent dose is 1.6 mg).
  • Table 3 shows the results of capillary electrophoretic separation of heat denatured citrate synthase protein aggregates after thermal treatment of the samples at 43 ° C for 70 minutes for other thymine derivatives. The experiments were otherwise performed as described above: rel. Protein efficacy in quantity precise. Compared to
  • the CCD-186Sk cystic fibrosis cell line (ATCC® CRL-1563 TM) was used.
  • the cystic fibrosis cell line CCD-186Sk homozygous carries the AF508 mutation affecting the CFTR gene.
  • the deletion of phenylalanine at position 508 of the CFTR gene is typical of this disease and affects over 70% of patients. Because of this deletion, the resulting CFTR protein is not completely correctly folded and for this reason fed to the cellular degradation processes and not transported - as the healthy protein - to the cell membrane and incorporated there as a transmembrane protein.
  • HSP27 plays a crucial role in the secretion of the Delta F508 CFTR protein towards degradation.
  • the experiments serve to demonstrate the hypothesis that Delta F508-CFTR protein, which is not secreted and degraded directly during production, is transported to the cell membrane where it regulates water and salt transport through the plasma membrane of the cells as a functioning transmembrane protein.
  • the CCD-186Sk cells are seeded for this purpose at a density of 100,000 cells / ml and with:
  • CFTR protein incubated as representatives of the purine derivatives in the respective concentration. Cells are harvested every 24 hours and it is checked if the treated cells present more CFTR on the cell surface than untreated cells.
  • an antibody is used that specifically labels the extracellular "loop" (amino acids 103-117) of the CFTR protein, which is labeled with the fluorescent dye FITC and detected by flow cytometry.
  • biotinylated HSP27 protein was first bound to super streptavidin sensors (SSA, Fortebio). Subsequently, the loaded sensors were incubated with Biocytin ("quenched"). For reference, uncharged SSA sensors were incubated with biocytin, and in the case of specific binding of the analyte to HSP27, the loaded HSP27 sensor produces a stronger signal than the response tracking reference sensor
  • the reaction buffer PBST 0.1% Tween 20 plus 3% DMSO
  • the association and then the dissociation of the respective analyte in real time are measured first. The analytes were measured in ascending concentrations: 1, 23 - 3.7 - 11, 11 - 33.33 - 100 - 300 ⁇ / ⁇ .
  • the cell line RPMI-8226 (DMSZ 402) is a "multiple myeloma" cell line and was chosen because bortezomib is used as the standard therapy for multiple myeloma. RPMI-8226 cells respond to bortezomib, express HSP27, and develop resistance to Velcade.
  • FIG. 4 a shows that the treatment with 0.75 ⁇ / kg accepromazine significantly inhibited the formation of resistance to the cytostatic bortezomib, and at the end of the experiment only 30,000 living cells per ml of medium were detectable.
  • RPMI 8226 cells treated with 30 ⁇ / L BVDU a final cell count of 160,000 cells / ml was detected, whereas the final cell count on culture without addition of an HSP27 inhibitor was 350,000 cells / ml.
  • Figure 4b At the same dosage, the test substances alone do not affect cell growth, since HSP27 is not needed in large quantities by the cancer cells without the pressure of the cytostatic.
  • Example 12 Inhibition of resistance formation of the tumor cell line RPMI-8226 by chlorpromazine
  • FIG. 5a shows that resistance to the cytostatic bortezomib was significantly inhibited by the treatment with 0.5 ⁇ mol / 1_ chlorpromazine and only 70,000 living cells per ml medium were detectable at the end of the experiment.
  • a final cell count of 160,000 cells / ml was detected for RPMI 8226 cells treated with 30 ⁇ / ⁇ _ BVDU, whereas the final cell count on culture without addition of an HSP27 inhibitor was 350,000 cells / ml.
  • RPMI 8226 cells were treated with the respective test substances alone, FIG. 5b. At the same dosage, the test substances alone do not affect cell growth.
  • Example 13 Inhibition of resistance formation of the tumor cell line RPMI-8226 by 2- (4-butylphenylamino) -9- (2-hydroxy-ethoxymethyl) -1,9-dihydro-purin-6-one
  • FIG. 6a shows that treatment with 1 ⁇ / L 2- (4-butylphenylamino) -9- (2-hydroxyethoxymethyl) -1,9-dihydro-purin-6-one results in the formation of resistance to the cytostatic bortezomib was significantly inhibited. At the end of the experiment were 240,000 living cells per ml of medium detectable. For the treatment with 30 ⁇ / L BVDU, a final cell count of 290,000 cells / ml was detected, whereas the final cell count on culture without addition of an HSP27 inhibitor amounts to 950,000 cells / ml. As a control, RPMI 8226 cells were treated with the respective test substances alone, FIG. 6b. At the same dosage, the test substances alone do not affect cell growth.
  • Example 14 Inhibition of resistance formation of the tumor cell line RPMI-8226 by 2- (3-trichlorovinyl-phenylamino) -1,9-dihydro-purin-6-ones
  • FIG. 7a shows that resistance to the cytostatic bortezomib was significantly inhibited by treatment with 1 ⁇ mol / 2 2- (3-trichlorovinyl-phenylamino) -1,9-dihydro-purin-6-one.
  • 270,000 live cells per ml of medium were detectable.
  • BVDU was detected to have a final cell count of 290,000 cells / ml, whereas the final cell count on culture without the addition of an HSP27 inhibitor was 950,000 cells / ml.
  • Figure 7b At the same dosage, the test substances alone do not affect cell growth.
  • Example 15 Inhibition of the resistance formation of the tumor cell line RPMI-8226 by 9H-xanthene-9-carboxylic acid [4- (5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl) -but- 2-enyl] -amide
  • FIG. 8a shows that treatment with 1 ⁇ mol / L 9H-xanthene-9-carboxylic acid [4- (5-methyl-2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-yl) -but -2-enyl] -amide resistance to the cytostatic bortezomib was significantly inhibited.
  • 290,000 live cells per ml of medium were detectable.
  • FIG. 9a shows that treatment with 1 ⁇ / L 2-biphenyl-4- yl-N- [4- (5-methyl-2,4-dioxo-3-dihydro-2H-pyrimidin-1-yl) -but-2-enyl] -acetam resistance to the cytostatic bortezomib was significantly inhibited. At the end of the experiment, 380,000 live cells per ml of medium were detectable.
  • the cell line U-937 is a histiocytic lymphoma (DSMZ ACC 5). This cell line also responds to treatment with bortezomib, expresses HSP27 and develops resistance to bortezomib. Bortezomib is not the standard therapy for this lymphoma but is discussed as a potential therapeutic.
  • FIG. 10a shows that resistance to the cytostatic bortezomib was significantly inhibited by the treatment with 0.75 ⁇ mol / L accepromazine. At the end of the experiment, 70,000 live cells per ml of medium were detectable. For the treatment with 30 ⁇ / L BVDU, a final cell count of 250,000 cells / ml was detected, whereas the final cell count during cultivation without the addition of an HSP27 inhibitor amounts to 450,000 cells / ml. For control U-937 cells were treated with the respective test substances alone, Figure 10b. At the same dosage, the test substances alone do not affect cell growth.
  • Example 18 Inhibition of resistance formation of the tumor cell line U-937 by 2- (4-butylphenylamino) -9- (2-hydroxy-ethoxymethyl) -1,9-dihydro-purin-6-one
  • FIG. 11a shows that treatment with 0.75 mol / L of 2- (4-butylphenylamino) -9- (2-hydroxy-ethoxymethyl) -1,9-dihydro-purin-6-one results in resistance formation the cytostatic bortezomib was significantly inhibited.
  • 30,000 live cells per ml of medium were detectable.
  • U-937 cells were treated with the respective test substances alone.
  • Figure 11 b At the same dosage, the test substances alone do not affect cell growth.

Landscapes

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

Abstract

La présente invention concerne de nouveaux inhibiteurs de HSP27, en particulier des dérivés de la thymine de la formule générale (VI), (VII) ou. (VIII) et des dérivés de la phénothiazine de la formule (V) et leur utilisation comme médicament pour l'inhibition sélective de la protéine de choc thermique HSP27 (HSPB1), en particulier pour une utilisation dans le traitement de carcinomes ou la mucoviscidose ; ces inhibiteurs dirigés contre HSP27 ont de façon particulièrement avantageuse une activité dans la gamme de concentrations de principe actif micromolaire inférieure ou sous-micromolaire.
EP15744197.3A 2014-07-28 2015-07-28 Dérivés de la thymine et de la chinazoline-dione pour inhiber hsp27 Withdrawn EP3174541A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE102014214798 2014-07-28
DE102014214799 2014-07-28
PCT/EP2015/067312 WO2016016268A1 (fr) 2014-07-28 2015-07-28 Dérivés de la thymine et de la chinazoline-dione pour inhiber hsp27

Publications (1)

Publication Number Publication Date
EP3174541A1 true EP3174541A1 (fr) 2017-06-07

Family

ID=53761383

Family Applications (2)

Application Number Title Priority Date Filing Date
EP15744196.5A Withdrawn EP3174545A1 (fr) 2014-07-28 2015-07-28 Inhibition efficace de hsp27
EP15744197.3A Withdrawn EP3174541A1 (fr) 2014-07-28 2015-07-28 Dérivés de la thymine et de la chinazoline-dione pour inhiber hsp27

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP15744196.5A Withdrawn EP3174545A1 (fr) 2014-07-28 2015-07-28 Inhibition efficace de hsp27

Country Status (4)

Country Link
US (2) US10940150B2 (fr)
EP (2) EP3174545A1 (fr)
CN (2) CN107073003A (fr)
WO (2) WO2016016267A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3819006A1 (fr) 2019-11-05 2021-05-12 Technische Universität Dresden Composés ayant un squelette de thymine à utiliser en médecine
CN111234247B (zh) * 2020-01-17 2021-08-27 福建师范大学 一种电致阻变的聚索烃型晶态材料的制备及在存储器中的应用
BR112023019511A2 (pt) * 2021-03-31 2023-10-31 Duke Street Bio Ltd Composto inibidor de parp7, composto, composição farmacêutica, kit farmacêutico para o tratamento de um câncer, método de tratamento de uma doença e/ou uma condição e/ou um distúrbio, e, método de síntese de um composto
WO2024015641A1 (fr) * 2022-07-15 2024-01-18 The Regents Of The University Of California Petite molécule pour le traitement du cancer de l'appendice

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012130166A1 (fr) * 2011-04-01 2012-10-04 Impact Therapeutics, Inc. 1-(arylméthyl)quinazoline-2,4(1h,3h)-diones en tant qu'inhibiteurs de parp et utilisation de celles-ci
US20120294846A1 (en) * 2011-05-12 2012-11-22 Oncogenex Technologies Inc. Treatment of Pulmonary and Pleural Fibrosis Using HSP27 Inhibitors
US20130150429A1 (en) * 2011-12-09 2013-06-13 The University Of British Columbia P27kip1 as a molecular marker for suitability and efficacy of treatment with hsp27 inhibitors

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5116823A (en) * 1990-02-27 1992-05-26 Roger Williams General Hospital Drug combinations containing AZT
US6995145B1 (en) * 1999-06-04 2006-02-07 Au Jessie L-S Methods and compositions for modulating drug activity through telomere damage
GB0003960D0 (en) 2000-02-18 2000-04-12 Pfizer Ltd Purine derivatives
AU2001247759A1 (en) 2000-03-24 2001-10-08 Duke University Characterization of grp94-ligand interactions and purification, screening, and therapeutic methods relating thereto
WO2004030660A2 (fr) 2002-10-02 2004-04-15 The University Of British Columbia Compositions et procedes pour le traitement du cancer de la prostate et d'autres types de cancer
CN1878556A (zh) 2003-09-18 2006-12-13 康宾纳特克斯公司 治疗肿瘤的联合药物
US20110070192A1 (en) 2003-11-14 2011-03-24 Bruno Tse Method of preparation of novel nucleoside analogs and uses
FR2876583B1 (fr) 2004-10-15 2007-04-13 Centre Nat Rech Scient Cnrse Utilisation de derives de purines pour la fabrication de medicaments pour le traitement de la mucoviscidose et de maladies liees a un defaut d'adressage des proteines dans les cellules
GB0509573D0 (en) 2005-05-11 2005-06-15 Merck Sharp & Dohme Therapeutic compounds
DE102005037733A1 (de) 2005-08-10 2007-02-15 Merck Patent Gmbh Adeninderivate
GB0519245D0 (en) 2005-09-20 2005-10-26 Vernalis R&D Ltd Purine compounds
AU2009243094B2 (en) 2006-11-06 2015-02-12 Massachusetts Institute Of Technology Immunomodulating compositions and methods of use thereof
GB0710277D0 (en) * 2007-05-30 2007-07-11 Univ Birmingham Use of antivirals in the treatment of medical disorders
DE102008030091B4 (de) 2008-06-25 2011-03-03 Resprotect Gmbh Uracilderivate und deren Verwendung
DE102008035299A1 (de) 2008-07-29 2010-02-04 Resprotect Gmbh Konjugat aus Hsp27 und einem Nukleosid, Verfahren zur Phosphorylierung von Hsp27 und Verwendung zur Herstellung eines Arzneimittels
WO2012099630A1 (fr) 2011-01-20 2012-07-26 University Of Rochester Compositions et méthodes de traitement ou de prévention d'une infection rétrovirale
US20120195911A1 (en) * 2011-02-01 2012-08-02 Artur Martynov Method of treatment of cancer patients
CA2829123C (fr) * 2011-03-14 2016-04-12 Eternity Bioscience Inc. Quinazolinediones et leur utilisation
AU2013216361B2 (en) 2012-02-02 2017-09-07 The University Of British Columbia Combination therapy for cancer using HSP27 inhibitor and EGFR tyrosine kinase inhibitors or anti-folates

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012130166A1 (fr) * 2011-04-01 2012-10-04 Impact Therapeutics, Inc. 1-(arylméthyl)quinazoline-2,4(1h,3h)-diones en tant qu'inhibiteurs de parp et utilisation de celles-ci
US20120294846A1 (en) * 2011-05-12 2012-11-22 Oncogenex Technologies Inc. Treatment of Pulmonary and Pleural Fibrosis Using HSP27 Inhibitors
US20130150429A1 (en) * 2011-12-09 2013-06-13 The University Of British Columbia P27kip1 as a molecular marker for suitability and efficacy of treatment with hsp27 inhibitors

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ORI KALID ET AL: "Small molecule correctors of F508del-CFTR discovered by structure-based virtual screening", JOURNAL OF COMPUTER-AIDED MOLECULAR DESIGN, KLUWER ACADEMIC PUBLISHERS, DO, vol. 24, no. 12, 26 October 2010 (2010-10-26), pages 971 - 991, XP019863450, ISSN: 1573-4951, DOI: 10.1007/S10822-010-9390-0 *
See also references of WO2016016268A1 *

Also Published As

Publication number Publication date
CN107074791A (zh) 2017-08-18
US20170216297A1 (en) 2017-08-03
CN107073003A (zh) 2017-08-18
US10940150B2 (en) 2021-03-09
WO2016016268A1 (fr) 2016-02-04
EP3174545A1 (fr) 2017-06-07
WO2016016267A1 (fr) 2016-02-04
US20180207160A1 (en) 2018-07-26

Similar Documents

Publication Publication Date Title
JP6949911B2 (ja) 標的化治療薬
JP2019135268A (ja) 準最適に投与された化学化合物の治療的有効性
DE60212836T2 (de) Pyridylpyrimidin-derivate als wirksame verbindungen gegen prionen-krankheiten
TWI791507B (zh) 使用ccr3-抑制劑治療老化相關損傷之方法及組合物
EP3684353A1 (fr) Formulations topiques de cannabinoïdes et leur utilisation dans le traitement de la douleur
CN106456795A (zh) 靶向治疗学
Hong et al. Pharmacokinetic modeling optimizes inhibition of the ‘undruggable’EWS-FLI1 transcription factor in Ewing Sarcoma
EP3174541A1 (fr) Dérivés de la thymine et de la chinazoline-dione pour inhiber hsp27
US20210085649A1 (en) Methods and use of compounds that bind to rela of nf-kb
CN102885818A (zh) 治疗或预防炎性痛的方法
EP3901151A1 (fr) Hétéroaryle halogéné et autres inhibiteurs de kinase hétérocycliques et leurs utilisations
US10765638B2 (en) Particle formulation with polycation complex
CN113939273A (zh) 用于药物递送的离子液体
CN113924295A (zh) 激酶抑制剂
CN1794986B (zh) 治疗癌症的新药物
CN114025766A (zh) 用于抑制gapdh的噁噻嗪化合物
Lee et al. Amlodipine besylate and amlodipine camsylate prevent cortical neuronal cell death induced by oxidative stress
Banerjee et al. A critical review of benzimidazole: Sky-high objectives towards the lead molecule to predict the future in medicinal chemistry
DE60214828T2 (de) Kombi-moleküle, welche eine inhibierende wirkung auf die signalübertragung und dna-schädigende eigenschaften haben
EP2699240B1 (fr) Dérivés de l'acadésine, produits et compositions les comprenant, leurs utilisations thérapeutiques et leurs procédés de synthèse
CN117545479A (zh) Pi3k抑制剂、纳米制剂及其用途
Borgini et al. Synthesis of Veliparib Prodrugs and Determination of Drug-Release-Dependent PARP-1 Inhibition
JP6796581B2 (ja) ミッシングタンパク質の小分子模倣体を用いた生理機能の回復
JP2024511466A (ja) Ａｌｋ－５阻害剤及びその使用
Aregbe et al. Population pharmacokinetic analysis of 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG) in adult patients with solid tumors

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

17P Request for examination filed

Effective date: 20170228

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

AX Request for extension of the european patent

Extension state: BA ME

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

17Q First examination report despatched

Effective date: 20180814

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

REG Reference to a national code

Ref country code: DE

Ref legal event code: R079

Free format text: PREVIOUS MAIN CLASS: A61K0031541500

Ipc: A61K0045060000

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

GRAJ Information related to disapproval of communication of intention to grant by the applicant or resumption of examination proceedings by the epo deleted

Free format text: ORIGINAL CODE: EPIDOSDIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

RIC1 Information provided on ipc code assigned before grant

Ipc: A61K 31/708 20060101ALI20200603BHEP

Ipc: A61P 35/00 20060101ALI20200603BHEP

Ipc: A61K 45/06 20060101AFI20200603BHEP

Ipc: A61K 31/513 20060101ALI20200603BHEP

Ipc: A61K 31/522 20060101ALI20200603BHEP

Ipc: A61K 31/5415 20060101ALI20200603BHEP

INTG Intention to grant announced

Effective date: 20200619

INTC Intention to grant announced (deleted)
GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20200813

GRAJ Information related to disapproval of communication of intention to grant by the applicant or resumption of examination proceedings by the epo deleted

Free format text: ORIGINAL CODE: EPIDOSDIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

INTC Intention to grant announced (deleted)
GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20210723

GRAJ Information related to disapproval of communication of intention to grant by the applicant or resumption of examination proceedings by the epo deleted

Free format text: ORIGINAL CODE: EPIDOSDIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

INTC Intention to grant announced (deleted)
GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20220103

GRAJ Information related to disapproval of communication of intention to grant by the applicant or resumption of examination proceedings by the epo deleted

Free format text: ORIGINAL CODE: EPIDOSDIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

INTC Intention to grant announced (deleted)
GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20230328

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20230808