DE102006008880A1 - aminopyrimidine derivatives - Google Patents

Abstract

Novel aminopyrimidine derivatives according to claim 1 are HSP90 inhibitors and can be used in the manufacture of a medicament for the treatment of diseases in which the inhibition, regulation and / or modulation of HSP90 plays a role.

Description

BACKGROUND THE INVENTION

Of the Invention was based on the object, new compounds with valuable Find properties, especially those for the production of medicines can be used.

The The present invention relates to compounds in which the inhibition, Regulation and / or modulation of HSP90 also plays a role pharmaceutical compositions containing these compounds, as well as the use of the compounds for the treatment of diseases, where HSP90 plays a role.

The proper folding and conformation of proteins in cells is done by ensured molecular chaperones and is critical for the regulation of the balance between protein synthesis and Degradation. Chaperones are important for the regulation of many central Functions of cells such as e.g. Cell proliferation and apoptosis (Jolly and Morimoto, 2000; Smith et al., 1998; Smith, 2001).

Heat shock proteins (heat shock proteins, HSPs)

The Tissues of a tissue respond to external stress, such as Heat, Hypoxia, oxidative stress, or toxins such as heavy metals or alcohols with the activation of a number of chaperones, which under the name "heat shock proteins "(HSPs) are known.

The Activation of HSPs protects the cell against injuries caused by such stressors accelerates the restoration of the physiological state and leads to a stress-tolerant condition of the cell.

Next this originally discovered by HSPs mediated protective mechanism in external Stress became more important chaperone functions over time for individual HSPs are also described under normal stress-free conditions. So For example, different HSPs regulate the correct folding, the intracellular Localization and function or the controlled removal of a series biologically important proteins of cells.

HSPs form a gene family with individual gene products whose cellular expression, Function and localization in different cells varies. The naming and classification within the family is due to their molecular weight e.g. HSP27, HSP70, and HSP90.

some Human diseases are based on incorrect protein folding (see review, e.g., Tytell et al., 2001; Smith et al., 1998). The Development of therapies resulting in the mechanism of chaperone-dependent protein folding engages, could therefore in such cases useful be. For example, lead in Alzheimer's disease, prion diseases or Huntington's Syndrome misfolded proteins to aggregate protein with neurodegenerative course. By wrong protein folding can also a loss of wild-type function arising from a dysregulated molecular and physiological Function can result.

HSPs is also considered a major importance in tumor diseases. There are e.g. Evidence that related to the expression of certain HSPs with the stage of progression of tumors (Martin et al., 2000; Conroy et al., 1996; Kawanishi et al., 1999; Jameel et al. 1992; Hoang et al., 2000; Lebeau et al., 1991).

The Fact that HSP90 is involved in several central oncogenic signaling pathways in the cell plays a role and certain natural substances with cancer-inhibiting activity Target HSP90 to the concept that inhibition of the function of HSP90 in the Treatment of tumor diseases would be useful.

One HSP90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17AAG), a derivative of geldanamycin is currently in clinical examination.

HSP90

HSP90 represents about 1-2% of the total cellular protein mass. It lies in the cell usually as a dimer and is associated with a variety of proteins, so-called co-chaperones (see, eg, Pratt, 1997). HSP90 is essential for the vitality of cells (Young et al., 2001) and plays a key role in the response to cellular stress by interacting with many proteins whose native folding has been altered by external stress, such as heat shock, to the original folding restore or prevent aggregation of the proteins (Smith et al., 1998).

It There is also evidence that HSP90 acts as a buffer against the effects of mutations has meaning, presumably through the correction incorrect protein folding caused by the mutation (Rutherford and Lindquist, 1998).

Furthermore HSP90 also has a regulatory significance. Under physiological Conditions plays HSP90, along with its homolog in endoplasmic Reticulum, GRP94, has a role in the cell balance to stabilize the conformation and maturing of various "client" key proteins to ensure. these can be divided into three groups: receptors for steroid hormones, Ser / Thr or Tyrosine kinases (e.g., ERBB2, RAF-1, CDK4, and LCK) and a library different proteins, e.g. mutant p53 or the catalytic Subunit of telomerase hTERT. Each of these proteins takes one key role in the regulation of physiological and biochemical processes of Cells.

The conserved human HSP90 family consists of four genes, the cytosolic HSP90α, the inducible HSP90β isoform (Hickey et al., 1989), GRP94 in the endoplasmic reticulum (Argon et al., 1999) and the HSP75 / TRAP1 in the mitochondrial Matrix (Felts et al., 2000). It is assumed that all members a similar one to the family Mode of action, but, depending on their location in the cell, on bind different "client" proteins. For example, ERBB2 is a specific "client" protein of GRP94 (Argon et al., 1999), while the type 1 tumor necrosis factor receptor (TNFR1) or retinoblastoma Protein (Rb) detected as "clients" of TRAP1 (Song et al., 1995; Chen et al., 1996).

HSP90 is on a series of complex interactions with a big one Number of "client" proteins and regulatory Involved proteins (Smith, 2001). Although precise molecular details still are not clear have biochemical experiments and investigations with the help of X-ray crystallography In recent years, details of the chaperone function of Decrypt HSP90 can (Prodromou et al., 1997; Stebbins et al., 1997). After that is HSP90 an ATP-dependent molecular chaperone (Prodromou et al, 1997), with dimerization important for the ATP hydrolysis is. The binding of ATP results in the formation a toroidal dimer structure, in which the two N-terminal Domains come in close contact with each other and a "switch" in the conformation cause. (Prodromou and Pearl, 2000).

Known HSP90 inhibitors

The first class of HSP90 inhibitors that were discovered were benzoquinone ansamycins with the compounds herbimycin A and geldanamycin. Originally became demonstrated with them the reversion of the malignant phenotype in fibroblasts, which had been induced by transformation with the v-Src oncogene (Uehara et al., 1985).

Later became a strong antitumoral activity in vitro (Schulte et al., 1998) and in vivo in animal models (Supko et al., 1995).

immunoprecipitation and investigations on affinity matrices then showed that the main mechanism of action of geldanamycin is a Binding to HSP90 (Whitesell et al., 1994; Schulte and Neckers, 1998). About that in addition, was determined by X-ray crystallographic Investigations revealed that geldanamycin centers around the ATP binding site competitively and inhibits the intrinsic ATPase activity of HSP90 (Prodromou et al., 1997; Panaretou et al., 1998). This is the origin of the multimeric HSP90 complex, with its characteristic as chaperone to act for "client" proteins, prevented. As a consequence, "client" proteins become via the ubiquitin-proteasome pathway reduced.

The Geldanamycin derivative 17-allylamino-17-demethoxygeldanamycin (17AAG) showed unchanged property in the inhibition of HSP90, the degradation of "client" proteins and antitumoral activity in cell cultures and in xenograft tumor models (Schulte et al, 1998; Kelland et al, 1999), but had significantly lower liver cytotoxicity than geldanamycin (Page et all 1997). 17AAG is currently in Phase I / II clinical Studies checked.

Radicicol, a macrocyclic antibiotic, also showed revision of the v-Src and v-Ha-Ras indu encoded malignant phenotype of fibroblasts (Kwon et al 1992, Zhao et al, 1995). Radicicol degrades a variety of signaling proteins as a consequence of HSP90 inhibition (Schulte et al., 1998). X-ray crystallographic studies showed that radicicol also binds to the N-terminal domain of HSP90 and inhibits intrinsic ATPase activity (Roe et al., 1998).

antibiotics The coumarin type is known to bind to the ATP binding site of the HSP90 homologs DNA gyrase in bacteria. Coumarin, Novobiocin, binds to the carboxy-terminal end of HSP90, that is, to another In HSP90, place as the benzoquinone ansamycins and radicicol which bind bind the N-terminal end of HSP90. (Marcu et al., 2000b).

The Inhibition of HSP90 by novobiocin results in degradation a big one Number of HSP90 dependent Signal proteins (Marcu et al., 2000a).

With PU3, a purine-derived HSP90 inhibitor, could cause degradation of signal proteins e.g. ERBB2, are shown. PU3 causes cell cycle arrest and differentiation in breast cancer cell lines (Chiosis et al., 2001).

HSP90 as a therapeutic target

By the involvement of HSP90 in the regulation of a large number of signaling pathways that are crucial to the phenotype of a Tumors have, and the discovery that certain natural substances their biological activity by inhibiting the activity of HSP90 HSP90 currently available as a new target for investigated the development of a tumor therapeutic (Neckers et al., 1999).

Of the Main mechanism of action of geldanamycin, 17AAG, and radicicol involves the inhibition of binding of ATP to the ATP binding site at the N-terminal End of the protein and the resulting inhibition of the intrinsic ATPase activity of HSP90 (see, e.g., Prodromou et al., 1997; Stebbins et al., 1997; Panaretou et al., 1998). Inhibition of ATPase activity of HSP90 prevents the recruitment of co-chaperones and favors the Formation of a HSP90 heterocomplex, the "client" protein via the ubiquitin-proteasome pathway the degradation supplies (See, e.g., Neckers et al., 1999; Kelland et al., 1999). The treatment of tumor cells with HSP90 inhibitors leads to selective degradation important proteins of fundamental importance for processes such as cell proliferation, Regulation of the cell cycle and apoptosis.

These Processes are common deregulated in tumors (see, e.g., Hostein et al., 2001).

A attractive rationale for The development of an inhibitor of HSP90 is that due to simultaneous degradation several proteins that interact with the transformed phenotype in the A strong tumor therapeutic effect achieved can be.

In particular, the present invention relates to compounds which inhibit, regulate and / or modulate HSP90, compositions containing these compounds and methods for their use in the treatment of HSP90-related diseases, such as tumor diseases, viral diseases such as hepatitis B (Waxman, 2002); Immunosuppression in transplantations (Bijlmakers, 2000 and Yorgin, 2000); Inflammatory Diseases (Bucci, 2000) such as Rheumatoid Arthritis, Asthma, Multiple Sclerosis, Type 1 Diabetes, Lupus Erythematosus, Psoriasis and Inflammatory Bowel Disease; Cystic fibrosis (Fuller, 2000); Diseases associated with angiogenesis (Hur, 2002 and Kurebayashi, 2001) such as diabetic retinopathy, hemangiomas, endometriosis and tumor angiogenesis; infectious diseases; Autoimmune diseases; ischemia; Promoting nerve regeneration (Rosen et al., WO 02/09696; Degranco et al., WO 99/51223; US 6,210,974 B1 ); fibrogenetic diseases such as scleroderma, polymyositis, systemic lupus, liver cirrhosis, keloid formation, interstitial nephritis and pulmonary fibrosis (Strehlow, WO 02/02123).

The The invention also relates to the use of the compounds according to the invention to protect normal cells against toxicity caused by chemo causing, as well as use in diseases, wherein protein misfolding or aggregation is a major causative factor, such as Scrapie, Creutzfeldt-Jakob Disease, Huntington or Alzheimer (Sittler, Hum. Mol. Genet., 10, 1307, 2001; Tratzelt et al., Proc. Nat. Acad. Sci., 92, 2944, 1995; Winklhofer et al., J. Biol. Chem., 276, 45160, 2001). A. Kamal et al. describe in Trends in Molecular Medicine, Vol. 6 June 2004, therapeutic and diagnostic applications of HSP90 activation, i.a. for treatment diseases of the central nervous system and cardiovascular diseases.

The Identification of small compounds that specifically inhibit HSP90, regulate and / or modulate is therefore desirable and an objective of present invention.

It was found that the Compounds of the invention and their salts with good compatibility possess very valuable pharmacological properties.

Especially show inhibitory properties of HSP90.

object The present invention therefore relates to the compounds of the invention as drugs and / or drugs in the treatment and / or prophylaxis of said diseases and use the compounds of the invention for the preparation of a pharmaceutical for the treatment and / or prophylaxis the said diseases as well as a method for the treatment of mentioned disorders comprising the administration of one or more inventive compounds to a patient in need of such administration.

Of the The host or patient may be of any mammalian species, e.g. A primate species, especially humans; Rodents, including mice, rats and hamsters; Rabbits; Horses, cattle, dogs, cats, etc. Animal models are for experimental studies of interest, where they are a model to treat a human disease.

WAS STANDING OF THE TECHNIQUE

Other Heteroaromatic compounds are described in WO 01/62233 A2 as adenosine receptor modulators.

In WO 01/72779 purine compounds are described, as well as their Use for the treatment of GRP94 (homolog or paralog to HSP90) -induced Diseases, such as tumors, where the cancerous tissue is a sarcoma or carcinoma selected from the group consisting of fibrosarcoma, myxosarcoma, liposarcoma, Chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, Lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing tumor, leiosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, Breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, Adenocarcinoma, sweat gland carcinoma, Sebaceous gland carcinoma, Papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, Bone marrow carcinoma, bronchogenic carcinoma, renal cell carcinoma, Hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonic Carcinoma, Wilms' tumor, Cervix cancer, Testicular tumor, Lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, Astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, Retinoblastoma, leukemia, Lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia and Heavy chain disease.

In WO 01/72779 is furthermore the use of the mentioned therein Discloses compounds for the treatment of viral diseases, wherein the viral pathogen is selected is from the group consisting of hepatitis type A, hepatitis type B, hepatitis type C, influenza, varicella, adenovirus, herpes simplex Type I (HSV-I), Herpes Simplex Type II (HSV-II), Rinderpest, Rhinovirus, Echovirus, Rotavirus, respiratory syncytial virus (RSV), papillomavirus, Papovavirus, cytomegalovirus, echinovirus, arbovirus, huntavirus, Coxsackie virus, mumps virus, measles virus, rubella virus, poliovirus, human immunodeficiency virus Type I (HIV-I) and Human Immunodeficiency Virus Type II (HIV-II).

In WO 01/72779 is also the use of the compounds mentioned there described for GRP94 modulation, wherein the modulated biological GRP94 activity an immune reaction in an individual, endoplasmic protein transport Reticulum, recovery from hypoxic / anoxic stress, recovery from malnutrition, Recovery from heat stress, or combinations thereof, causes, and / or wherein the disorder a type of cancer is an infectious disease, a disorder that with a disturbed Protein transport from the endoplasmic reticulum, a disorder that with ischemia / reperfusion or combinations thereof, those with ischemia / reperfusion accompanying disorder a consequence of cardiac arrest, asystole and delayed ventricular arrhythmias, Heart surgery, cardiopulmonary Bypass surgery, organ transplantation, spinal cord injury, head trauma, Stroke, thromboembolic stroke, haemorrhagic stroke, cerebral vasospasm, hypotension, hypoglycemia, status epilepticus, an epileptic Seizure, anxiety, schizophrenia, a neurodegenerative disorder, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS) or stress when newborn is.

In WO 01/72779 is finally the use of an effective amount of a GRP94 protein modulator for the manufacture of a medicament, for modifying a subsequent cellular Reaction to an ischemic Condition at a tissue site in an individual, by treatment of cells at the tissue site with the GRP94 protein modulator, thus the GRP94 activity in cells is so intensified, that a subsequent cellular reaction to an ischemic Condition changed is, with the subsequent ischemic Condition preferably the consequence of cardiac arrest, asystole and delayed ventricular Arrhythmias, heart surgery, cardiopulmonary bypass surgery, organ transplantation, spinal cord injury, Head trauma, stroke, thromboembolic stroke, hemorrhagic stroke, cerebral vasospasm, hypotension, hypoglycemia, status epilepticus, one epileptic seizure, anxiety, schizophrenia, a neurodegenerative disorder Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS) or stress in the newborn, or where the tissue site is the donor tissue for one Transplantation is.

• Argon Y and Simen BB. 1999 "Grp94, an ER chaperone with protein and peptide binding properties", Semin. Cell Dev. Biol., Vol. 10, pp. 495–505.
• Bijlmakers M-JJE, Marsh M. 2000 "Hsp90 is essential for the synthesis and subsequent membrane association, but not the maintenance, of the Src-kinase p56lck", Mol. Biol. Cell, Vol. 11(5), pp. 1585–1595.
• Bucci M; Roviezzo F; Cicala C; Sessa WC, Cirino G. 2000 "Geldanamycin, an inhibitor of heat shock protein 90 (Hsp90) mediated signal transduction has anti-inflammatory effects and interacts with glucocorticoid receptor in vivo", Brit. J. Pharmacol., Vol 131(1), pp. 13–16.
• Carreras CW, Schirmer A, Zhong Z, Santi VS. 2003 "Filter binding assay for the geldanamycin-heat shock protein 90 interaction", Analytical Biochem., Vol 317, pp 40–46.
• Chen C-F, Chen Y, Dai KD, Chen P-L, Riley DJ and Lee W-H. 1996 "A new member of the hsp90 family of molecular chaperones interacts with the retinoblastoma protein during mitosis and after heat shock", Mol. Cell. Biol., Vol. 16, pp. 4691–4699.
• Chiosis G, Timaul MN, Lucas B, Munster PN, Zheng FF, Sepp-Lozenzino L and Rosen N. 2001 "A small molecule designed to bind to the adenine nucleotide pocket of HSP90 causes Her2 degradation and the growth arrest and differentiation of breast cancer cells", Chem. Biol., Vol. 8, pp. 289–299.
• Chiosis G, Lucas B, Shtil A, Huezo H, Rosen N 2002 "Development of a purine-scaffold novel class of HSP90 binders that inhibit the proliferation of cancer cells and induce the degradation of her2 tyrosine kinase". Bioorganic Med. Chem, Vol 10, pp 3555–3564.
• Conroy SE and Latchman DS. 1996 "Do heat shock proteins have a role in breast cancer?", Brit. J. Cancer, Vol. 74, pp. 717–721.
• Felts SJ, Owen BAL, Nguyen P, Trepel J, Donner DB and Toft DO. 2000 "The HSP90-related protein TRAP1 is a mitochondrial protein with distinct functional properties", J. Biol. Chem, Vol. 5, pp. 3305–3312.
• Fuller W, Cuthbert AW. 2000 "Post-translational disruption of the delta F508 cystic fibrosis transmembrane conductance regulator (CFTR)-molecular Chaperone complex with geldanamycin stabilizes delta F508 CFTR in the rabbit reticulocyte lysate", J. Biol. Chem, Vol. 275(48), pp. 37462–37468.
• Hickey E, Brandon SE, Smale G, Lloyd D and Weber LA. 1999 "Sequence and regulation of a gene encoding a human 89-kilodalton heat shock protein", Mol. Cell. Biol., Vol. 9, pp. 2615–2626.
• Hoang AT, Huang J, Rudra-Gonguly N, Zheng J, Powell WC, Rabindron SK, Wu C and Roy-Burman P. 2000 "A novel association between the human heat shock transcription factor 1 (HSF1) and prostate adenocarcinoma, Am. J. Pathol., Vol. 156, pp. 857–864.
• Hostein I, Robertson D, Di Stefano F, Workman P and Clarke PA. 2001 "Inhibition of signal transduction by the HSP90 inhibitor 17-allylamino-17-demethoxygeldanamycin results in cytostasis and apoptosis", Cancer Res., Vol. 61, pp. 4003–4009.
• Hur E, Kim H-H, Choi SM, Kim JH, Yim S, Kwon HJ, Choi Y, Kim DK, Lee M-O, Park H. 2002 "Reduction of hypoxia-induced transcription through the repression of hypoxia-inducible factor-1α/aryl hydrocarbon receptor nuclear translocator DNA binding by the 90-kDa heat-shock protein inhibitor radicicol", Mol. Pharmacol., Vol 62(5), pp. 975–982.
• Jameel A, Skilton RA, Campbell TA, Chander SK, Coombes RC and Luqmani YA. 1992 "Clinical
• Jolly C and Morimoto RI. 2000 "Role of the heat shock response and molecular chaperones in oncogenesis and cell death", J. Natl. Cancer Inst., Vol. 92, pp. 1564–1572.
• Kawanishi K, Shiozaki H, Doki Y, Sakita I, Inoue M, Yano M, Tsujinata T, Shamma A and Monden M. 1999 "Prognostic significance of heat shock proteins 27 and 70 in patients with squamous cell carcinoma of the esophagus", Cancer, Vol. 85, pp. 1649–1657.
• Kelland LR, Abel G, McKeage MJ, Jones M, Goddard PM, Valenti M, Murrer BA, and Harrap KR. 1993 "Preclinical antitumour evaluation of bis-acetalo-amino-dichloro-cyclohexylamine platinum (IV): an orally active platinum drug", Cancer Research, Vol. 53, pp. 2581–2586.
• Kelland LR, Sharp SY, Rogers PM, Myers TG and Workman P. 1999 "DT-diaphorase expression and tumor cell sensitivity to 17-allylamino, 17-demethoxygeldanamycin, an inhibitor of heat shock protein 90", J. Natl. Cancer Inst., Vol. 91, pp. 1940–1949.
• Kurebayashi J, Otsuki T, Kurosumi M, Soga S, Akinaga S, Sonoo, H. 2001 "A radicicol derivative, KF58333, inhibits expression of hypoxia-inducible factor-1α and vascular endothelial growth factor, angiogenesis and growth of human breast cancer xenografts", Jap. J. Cancer Res., Vol. 92(12), 1342–1351.
• Kwon HJ, Yoshida M, Abe K, Horinouchi S and Bepple T. 1992 "Radicicol, an agent inducing the reversal of transformed phentoype of src-transformed fibroblasts, Biosci., Biotechnol., Biochem., Vol. 56, pp. 538–539.
• Lebeau J, Le Cholony C, Prosperi MT and Goubin G. 1991 "Constitutive overexpression of 89 kDa heat shock protein gene in the HBL100 mammary cell line converted to a tumorigenic phenotype by the EJE24 Harvey-ras oncogene", Oncogene, Vol. 6, pp. 1125–1132.
• Marcu MG, Chadli A, Bouhouche I, Catelli M and Neckers L. 2000a "The heat shock protein 90 antagonist novobiocin interacts with a previously unrecognized ATP-binding domain in the carboxyl terminus of the chaperone", J. Biol. Chem., Vol. 275, pp. 37181–37186.
• Marcu MG, Schulte TW and Neckers L. 2000b "Novobiocin and related coumarins and depletion of heat shock protein 90-dependent signaling proteins", J. Natl. Cancer Inst., Vol. 92, pp. 242–248.
• Martin KJ, Kritzman BM, Price LM, Koh B, Kwan CP, Zhang X, MacKay A, O'Hare MJ, Kaelin CM, Mutter GL, Pardee AB and Sager R. 2000 "Linking gene expression patterns to therapeutic groups in breast cancer", Cancer Res., Vol. 60, pp. 2232–2238.
• Neckers L, Schulte TW and Momnaaugh E. 1999 "Geldanamycin as a potential anti-cancer agent: its molecular target and biochemical activity", Invest. New Druqs, Vol. 17, pp. 361–373.
• Page J, Heath J, Fulton R, Yalkowsky E, Tabibi E, Tomaszewski J, Smith A and Rodman L. 1997 "Comparison of geldanamycin (NSC-122750) and 17-allylaminogeldanamycin (NSC-330507D) toxicity in rats", Proc. Am. Assoc. Cancer Res., Vol. 38, pp. 308.
• Panaretou B, Prodromou C, Roe SM, OBrien R, Ladbury JE, Piper PW and Pearl LH. 1998 "ATP binding and hydrolysis are essential to the function of the HSP90 molecular chaperone in vivo", EMBO J., Vol. 17, pp. 4829–4836.
• Pratt WB. 1997 "The role of the HSP90-based chaperone system in signal transduction by nuclear receptors and receptors signalling via MAP kinase", Annu. Rev. Pharmacol. Toxicol., Vol. 37, pp. 297–326.
• Prodromou C, Roe SM, O'Brien R, Ladbury JE, Piper PW and Pearl LH. 1997 "Identification and structural characterization of the ATP/ADP-binding site in the HSP90 molecular chaperone", Cell, Vol. 90, pp. 65–75.
• Prodromou C, Panaretou B, Chohan S, Siligardi G, O'Brien R, Ladbury JE, Roe SM, Piper PW and Pearl LH. 2000 "The ATPase cycle of HSP90 drives a molecular "clamp" via transient dimerization of the N-terminal domains", EMBO J., Vol. 19, pp. 4383–4392.
• Roe SM, Prodromou C, O'Brien R, Ladbury JE, Piper PW and Pearl LH. 1999 "Structural basis for inhibition of the HSP90 molecular chaperone by the antitumour antibiotics radicicol and geldanamycin", J. Med. Chem., Vol. 42, pp. 260–266.
• Rutherford SL and Lindquist S. 1998 "HSP90 as a capacitor for morphological evolution. Nature, Vol. 396, pp. 336–342.
• Schulte TW, Akinaga S, Murakata T, Agatsuma T, Sugimoto S, Nakano H, Lee YS, Simen BB, Argon Y, Felts S, Toft DO, Neckers LM and Sharma SV. 1999 "Interaction of radicicol with members of the heat shock protein 90 family of molecular chaperones", Mol. Endocrinoloqy, Vol. 13, pp. 1435–1448.
• Schulte TW, Akinaga S, Soga S, Sullivan W, Sensgard B, Toft D and Neckers LM. 1998 "Antibiotic radicicol binds to the N-terminal domain of HSP90 and shares important biologic activities with geldanamcyin", Cell Stress and Chaperones, Vol. 3, pp. 100–108.
• Schulte TW and Neckers LM. 1998 "The benzoquinone ansamycin 17-allylamino-17-demethoxygeldanamcyin binds to HSP90 and shares important biologic activities with geldanamycin", Cancer Chemother. Pharmacol., Vol. 42, pp. 273–279.
• Smith DF. 2001 "Chaperones in signal transduction", in: Molecular chaperones in the cell (P Lund, ed.; Oxford University Press, Oxford and NY), pp. 165–178.
• Smith DF, Whitesell L and Katsanis E. 1998 "Molecular chaperones: Biology and prospects for pharmacological intervention", Pharmacological Reviews, Vol. 50, pp. 493–513.
• Song HY, Dunbar JD, Zhang YX, Guo D and Donner DB. 1995 "Identification of a protein with homology to hsp90 that binds the type 1 tumour necrosis factor receptor", J. Biol. Chem., Vol. 270, pp. 3574–3581.
• Stebbins CE, Russo A, Schneider C, Rosen N, Hartl FU and Pavletich NP. 1997 "Crystal structure of an HSP90-geldanamcyin complex: targeting of a protein chaperone by an antitumor agent", Cell, Vol. 89, pp. 239–250.
• Supko JG, Hickman RL, Grever MR and Malspeis L. 1995 "Preclinical pharmacologic evaluation of geldanamycin as an antitumour agent", Cancer Chemother. Pharmacol., Vol. 36, pp. 305–315.
• Tytell M and Hooper PL. 2001 "Heat shock proteins: new keys to the development of cytoprotective therapies", Emerging Therapeutic Targets, Vol. 5, pp. 267–287.
• Uehara U, Hori M, Takeuchi T and Umezawa H. 1986 "Phenotypic change from transformed to normal induced by benzoquinoid ansamycins accompanies inactivation of p6Osrc in rat kidney cells infected with Rous sarcoma virus", Mol. Cell. Biol., Vol. 6, pp. 2198–2206.
• Waxman, Lloyd H. Inhibiting hepatitis C virus processing and replication. (Merck & Co., Inc., USA). PCT Int. Appl. (2002), WO 0207761 Whitesell L, Mimnaugh EG, De Costa B, Myers CE and Neckers LM. 1994 "Inhibition of heat shock protein HSP90-pp60v-src heteroprotein complex formation by benzoquinone ansamycins: essential role for stress proteins in oncogenic transformation", Proc. Natl. Acad. Sci. USA., Vol. 91, pp. 8324–8328.
• Yorgin et al. 2000 "Effects of geldanamycin, a heat-shock protein 90-binding agent, on T cell function and T cell nonreceptor protein tyrosine kinases", J. Immunol., Vol 164(6), pp. 2915–2923.
• Young JC, Moarefi I and Hartl FU. 2001 "HSP90: a specialized but essential protein-folding tool", J. Cell. Biol., Vol. 154, pp. 267–273.
• Zhao JF, Nakano H and Sharma S. 1995 "Suppression of RAS and MOS transformation by radicicol", Oncoqene, Vol. 11, pp. 161–173.

Further literature:

• Argon Y and Simen BB. 1999 "Grp94, to ER chaperones with protein and peptide binding properties", Semin. Cell Dev. Biol., Vol. 10, pp. 495-505.
• Bijlmakers M-JJE, Marsh M 2000 "Hsp90 is essential for the synthesis and subsequent membrane association, but not the maintenance, of the Src kinase p56lck", Mol. Biol. Cell, Vol. 11 (5), pp. From 1585 to 1595.
• Bucci M; Roviezzo F; Cicala C; Sessa WC, Cirino G. 2000 "Geldanamycin, inhibitor of heat shock protein 90 (Hsp90) mediated signal transduction has anti-inflammatory effects and interact with glucocorticoid receptor in vivo", Brit. J. Pharmacol., Vol 131 (1), pp. 13-16.
• Carreras CW, Schirmer A, Zhong Z, Santi VS. 2003 "Filter binding assay for the geldanamycin-heat shock protein 90 interaction", Analytical Biochem., Vol. 317, pp. 40-46.
• Chen CF, Chen Y, Dai KD, Chen PL, Riley DJ and Lee WH. 1996 "A new member of the hsp90 family of molecular chaperones interacts with the retinoblastoma protein during mitosis and after heat shock", Mol. Cell. Biol., Vol. 16, pp. 4,691 to 4,699.
• Chiosis G, Timaul MN, Lucas B, Munster PN, Zheng FF, Sepp-Lozenzino L and Rosen N. 2001 "A small molecule designed to bind to the adenine nucleotide pocket of HSP90 causes her2 degradation and the growth arrest and differentiation of breast cancer cells ", Chem. Biol., Vol. 8, pp. 289-299.
• Chiosis G, Lucas B, Shtil A, Huezo H, Rosen N 2002 "Development of a purine-scaffold novel class of HSP90 binders that inhibit the proliferation of cancer cells and induce the degradation of her2 tyrosine kinase". Bioorganic Med. Chem, Vol. 10, pp 3555-3564.
• Conroy SE and Latchman DS. 1996 "Do heat shock proteins have a role in breast cancer?", Brit. J. Cancer, Vol. 74, pp. 717-721.
• Felts SJ, Owen BAL, Nguyen P, Trepel J, Thunder DB and Toft DO. 2000 "The HSP90-related protein TRAP1 is a mitochondrial protein with distinct functional properties", J. Biol. Chem., Vol. 5, pp. 3305-3312.
• Fuller W, Cuthbert AW. 2000 "Post-translational disruption of the delta F508 cystic fibrosis transmembrane conductance regulator (CFTR) -molecular chaperone complex with geldanamycin-stabilizes delta F508 CFTR in the rabbit reticulocyte lysate", J. Biol. Chem., Vol. 275 (48), pp. From 37,462 to 37,468.
• Hickey E, Brandon SE, Smale G, Lloyd D and Weber LA. 1999 "Sequence and regulation of a gene encoding a human 89-kilodalton heat shock protein", Mol. Cell. Biol., Vol. 9, pp. 2615-2626.
• Hoang AT, Huang J, Rudra-Gonguly N, Zheng J, Powell WC, Rabindron SK, Wu C and Roy-Burman P. 2000 "A novel association between the human heat shock transcription factor 1 (HSF1) and prostate adenocarcinoma, Am. J. Pathol., Vol. 156, pp. 857-864.
• Hostein I, Robertson D, Di Stefano F, Workman P and Clarke PA. 2001 "Inhibition of signal transduction by the HSP90 inhibitor 17-allylamino-17-demethoxygeldanamycin results in cytostasis and apoptosis", Cancer Res., Vol. 61, pp. 4003 to 4009.
• Hur E, Kim HH, Choi SM, Kim JH, Yim S, Kwon HJ, Choi Y, Kim DK, Lee MO, Park H. 2002 "Reduction of hypoxia-induced transcription through the repression of hypoxia-inducible factor-1α / aryl hydrocarbon receptor nuclear translocator DNA binding by the 90-kDa heat-shock protein inhibitor radicicol ", Mol. Pharmacol., Vol. 62 (5), pp. 975-982.
• Jameel A, Skilton RA, Campbell TA, Chander SK, Coombes RC and Luqmani YA. 1992 "Clinical
• Jolly C and Morimoto RI. 2000 "Role of the heat shock response and molecular chaperones in oncogenesis and cell death", J. Natl. Cancer Inst., Vol. 92, pp. From 1564 to 1572.
• Kawanishi K, Shiozaki H, Doki Y, Sakita I, Inoue M, Yano M, Tsujinata T, Shamma A and Monden M 1999 "Prognostic significance of heat shock proteins 27 and 70 in patients with squamous cell carcinoma of the esophagus", Cancer , Vol. 85, pp. 1649-1657.
• Kelland LR, Abel G, McKeage MJ, Jones M, Goddard PM, Valenti M, Murrer BA, and Harrap KR. 1993 "Preclinical antitumour evaluation of bis-aceto-amino-dichloro-cyclohexylamine platinum (IV): on orally active platinum drug", Cancer Research, Vol. 53, pp. 2581-2586.
• Kelland LR, Sharp SY, Rogers PM, Myers TG and Workman P. 1999 "DT-diaphorase expression and tumor cell sensitivity to 17-allylamino, 17-demethoxygeldanamycin, to inhibitor of heat shock protein 90", J. Natl. Cancer Inst., Vol. 91, pp. 1940-1949.
• Kurebayashi J, Otsuki T, Kurosumi M, Soga S, Akinaga S, Sonoo, H., 2001. A radicicol derivative, KF58333, inhibits expression of hypoxia-inducible factor-1α and vascular endothelial growth factor, angiogenesis and growth of human breast cancer xenografts ", Jap. J. Cancer Res., Vol. 92 (12), 1342-1351.
• Kwon HJ, Yoshida M, Abe K, Horinouchi S and Bepple T. 1992 "Radicicol, to the inducer of the reversal of transformed phentoype of src-transformed fibroblasts, Biosci., Biotechnol., Biochem., Vol. 56, pp. 538- 539th
• Lebeau J, Le Cholony C, Prosperi MT and Goubin G. 1991 "Constitutive overexpression of 89 kDa heat shock protein gene in the HBL100 mammary cell line transformed into a tumorigenic phenotype by the EJE24 Harvey-ras oncogene", Oncogene, Vol. pp. From 1125 to 1132.
• Marcu MG, Chadli A, Bouhouche I, Catelli M and Necker's L. 2000a "The heat shock protein 90 antagonist novobiocin interacts with a previously unrecognized ATP-binding domain in the carboxyl terminus of the chaperone", J. Biol. Chem., Vol 275, pp. 37181-37186.
• Marcu MG, Schulte TW and Neckers L 2000b "Novobiocin and related coumarin and depletion of heat shock protein 90-dependent signaling proteins", J. Natl. Cancer Inst., Vol. 92, pp. 242-248.
• Martin KJ, Kritzman BM, Price LM, Koh B, Kwan CP, Zhang X, MacKay A, O'Hare MJ, Kaelin CM, Mother GL, Pardee AB and Sager R. 2000 "Linking gene expression patterns to therapeutic groups in breast cancer , Cancer Res., Vol. 60, pp. From 2232 to 2238.
• Neckers L, Schulte TW and Momnaaugh E. 1999 "Geldanamycin as a potential anti-cancer agent: its molecular target and biochemical activity", Invest. New Druqs, Vol. 17, pp. 361-373.
• 1997, "Comparison of geldanamycin (NSC-122750) and 17-allylaminogeldanamycin (NSC-330507D) toxicity in rats," Proc. J, Heath J, Fulton R, Yalkowsky E, Tabibi E, Tomaszewski J, Smith A, and Rodman L. At the. Assoc. Cancer Res., Vol. 38, pp. 308th
• Panaretou B, Prodromou C, Roe SM, OBrien R, Ladbury JE, Piper PW and Pearl LH. 1998 "ATP binding and hydrolysis are essential to the function of the HSP90 molecular chaperone in vivo", EMBO J., Vol. 17, pp. 4829 to 4836.
• Pratt WB. 1997 "The role of the HSP90-based chaperone system in signal transduction by nuclear receptors and receptors signaling via MAP kinase", Annu. Rev. Pharmacol. Toxicol., Vol. 37, pp. 297-326.
• Prodromou C, Roe SM, O'Brien R, Ladbury JE, Piper PW and Pearl LH. 1997 "Identification and structural characterization of the ATP / ADP binding site in the HSP90 molecular chaperone", Cell, Vol. 90, pp. 65-75.
• Prodromou C, Panaretou B, Chohan S, Siligardi G, O'Brien R, Ladbury JE, Roe SM, Piper PW and Pearl LH. 2000 "The ATPase cycle of HSP90 drives a molecular" clamp "via transient dimerization of the N-terminal domains", EMBO J., Vol. 19, pp. From 4383 to 4392.
• Roe SM, Prodromou C, O'Brien R, Ladbury JE, Piper PW and Pearl LH. 1999 "Structural base for inhibition of the HSP90 molecular chaperone by the antitumour antibiotics radicicol and geldanamycin", J. Med. Chem., Vol. 42, pp. 260-266.
• Rutherford SL and Lindquist S. 1998 "HSP90 as a Condenser for Morphological Evolution. Nature, Vol. 396, pp. 336-342.
• Schulte TW, Akinaga S, Murakata T, Agatsuma T, Sugimoto S, Nakano H, Lee YS, Simen BB, Argon Y, Felts S, Toft DO, Neckers LM and Sharma SV. 1999 "Interaction of radicicol with members of the heat shock protein 90 family of molecular chaperones", Mol. Endocrinoloy, Vol. 13, pp. 1435-1448.
• Schulte TW, Akinaga S, Soga S, Sullivan W, Sensgard B, Toft D and Neckers LM. 1998 "Antibiotic radicicol binds to the N-terminal domain of HSP90 and shares important biological activities with geldanamcyin", Cell Stress and Chaperones, Vol. 3, pp. 100-108.
• Schulte TW and Neckers LM. 1998 "The benzoquinone ansamycin 17-allylamino-17-demethoxygeldanamcyin binds to HSP90 and shares important biologic activities with geldanamycin", Cancer Chemother. Pharmacol., Vol. 42, pp. 273-279.
• Smith DF. 2001 "Chaperones in signal transduction", in: Molecular chaperones in the cell (P Lund, ed., Oxford University Press, Oxford and NY), pp. 165-178.
• Smith DF, Whitesell L and Katsanis E. 1998 "Molecular chaperones: Biology and Prospects for Pharmacological Intervention", Pharmacological Reviews, Vol. 493-513.
• Song HY, Dunbar JD, Zhang YX, Guo D and Thunder DB. 1995 "Identification of a protein with homology to hsp90 that binds the type 1 tumor necrosis factor receptor", J. Biol. Chem., Vol. 270, pp. 3574 to 3581.
• Stebbins CE, Russo A, Schneider C, Rosen N, Hartl FU and Pavletich NP. 1997 "Crystal structure of HSP90 geldanamcyin complex: targeting a protein chaperone by an antitumor agent", Cell, Vol. 89, pp. 239-250.
• Supko JG, Hickman RL, Grever MR and Malspeis L. 1995 "Preclinical pharmacologic evaluation of geldanamycin as an antitumour agent", Cancer Chemother. Pharmacol., Vol. 36, pp. 305-315.
• Tytell M and Hooper PL. 2001 "Heat shock proteins: new keys to the development of cytoprotective therapies", Emerging Therapeutic Targets, Vol. 5, pp. 267-287.
• Uehara U, Hori M, Takeuchi T and Umezawa H. 1986 "Phenotypic change from normal to normal induced by benzoquinoid ansamycin accompanies inactivation of p6Osrc in rat kidney cells infected with Rous sarcoma virus", Mol. Cell. Biol., Vol. 6, pp. From 2198 to 2206.
• Waxman, Lloyd H. Inhibiting hepatitis C virus processing and replication. (Merck & Co., Inc., USA). PCT Int. Appl. (2002), WO 0207761 Whitesell L, Mimnaugh EG, De Costa B, Myers CE and Neckers LM. 1994 "Inhibition of heat shock protein HSP90-pp60v-src heteroprotein complex formation by benzoquinone ansamycin: essential role for stress proteins in oncogenic transformation", Proc. Natl. Acad. Sci. USA., Vol. 91, pp. From 8324 to 8328.
• Yorgin et al. 2000 "Effects of geldanamycin, a heat-shock protein 90-binding agent, on cell function and T cell non-receptor protein tyrosine kinases", J. Immunol., Vol 164 (6), pp. 2915-2923.
• Young JC, Moarefi I and Hartl FU. 2001 "HSP90: a specialized but essential protein-folding tool", J. Cell. Biol., Vol. 154, pp. 267-273.
• Zhao JF, Nakano H and Sharma p. 1995 "Suppression of RAS and MOS transformation by radicicol", Oncoqene, Vol. 11, pp. 161-173.

SUMMARY THE INVENTION

sowie ihre pharmazeutisch verwendbaren Derivate, Salze, Solvate, Tautomere und Stereoisomere, einschließlich deren Mischungen in allen Verhältnissen.The invention relates to compounds selected from the group

and their pharmaceutically usable derivatives, salts, solvates, tautomers and stereoisomers, including mixtures thereof in all ratios.

object The invention also relates to the hydrates and solvates of these compounds. Among solvates of the compounds are additions of inert solvent molecules to the Compounds understood because of their mutual attraction form. Solvates are e.g. Mono or dihydrate or alcoholates.

Under pharmaceutically usable derivatives are understood e.g. the salts the compounds of the invention as well as so-called prodrug compounds.

Under Prodrug derivatives are understood with z. B. alkyl or acyl groups, Sugars or oligopeptides modified inventive compounds, in the organism rapidly to the effective compounds of the invention be split.

For this belong also biodegradable polymer derivatives of the compounds according to the invention, as this z. In Int. J. Pharm. 115, 61-67 (1995).

Of the Expression "effective Quantity "means the amount of a drug or pharmaceutical agent, the one biological or medical answer in a tissue, System, animal or human, e.g. from a researcher or doctors are sought or desired.

Moreover, the term "therapeutically effective amount" means an amount that, as compared to a corresponding subject who has not received that amount, results in:
improved curative treatment, cure, prevention or elimination of a disease, a clinical picture, a disease state, a condition, a disorder or side effects or even a reduction in the progression of a disease, a condition or a disorder.

The Term "therapeutic effective amount "also includes the amounts being effective, the normal physiological function to increase.

object The invention also relates to mixtures of the compounds according to the invention, e.g. Mixtures of two diastereomers, e.g. in the ratio 1: 1, 1: 2, 1: 3, 1: 4, 1: 5, 1:10, 1: 100 or 1: 1000.

Especially These are preferably mixtures of stereoisomeric compounds.

The compounds may possess one or more chiral centers and therefore in different ste reoisomeric forms occur.

The Compounds of the invention and also the starting materials for their preparation are in the rest methods known per se, as described in the literature (e.g., in standard works such as Houben-Weyl, Methods of Organic Chemistry, Georg Thieme Verlag, Stuttgart) are described, namely under reaction conditions suitable for the reactions mentioned are known and suitable. It can also be known by itself, not closer here mentioned Make use of variants.

The Starting materials can, if desired, also be formed in situ so that they can be removed from the reaction mixture not isolated, but immediately further to the compounds of the invention implements.

The Starting compounds are generally known. Are they new, like that can but they are prepared by methods known per se.

The Compounds "A1" - "A13" according to the invention may preferably can be obtained by e.g. Reacting chloropyrimidine derivatives with arylboronic acids (Suzuki-coupling).

The Implementation is carried out by methods known to the person skilled in the art. Lit .: J.M. Shomak and T.J. Delia, J. Org. Chem. 2001, 66, 7125-7128.

The Reaction takes place in a suitable inert solvent.

When inert solvent are suitable e.g. Hydrocarbons such as hexane, petroleum ether, benzene, Toluene or xylene; chlorinated hydrocarbons such as trichlorethylene, 1,2-dichloroethane, carbon tetrachloride, chloroform or dichloromethane; Alcohols such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; Ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran (THF) or dioxane; Glycol ethers such as ethylene glycol monomethyl or monoethyl ether (methyl glycol or ethyl glycol), ethylene glycol dimethyl ether (Diglyme); Ketones such as acetone or butanone; Amides such as acetamide, dimethylacetamide or dimethylformamide (DMF); Nitriles such as acetonitrile; sulfoxides such as dimethyl sulfoxide (DMSO); Carbon disulphide; Carboxylic acids like formic acid or acetic acid; Nitrover compounds such as nitromethane or nitrobenzene; Esters such as ethyl acetate or mixtures of the solvents mentioned.

The Response time varies depending on the conditions used a few minutes and 14 days, the reaction temperature between about -30 ° and 140 °, normally between -10 ° and 130 °, in particular between about 30 ° and about 125 °.

The compounds of the invention "A14" - "A16" can preferably be obtained by reacting 2-amino-4-hydroxy-5-cyanpyrimidinderivate with POCl ₃ under standard conditions.

Pharmaceutical salts and other forms

The abovementioned compounds according to the invention can be used in their final non-salt form. On the other hand, the present invention also encompasses the use of these compounds in the form of their pharmaceutically acceptable salts, which can be derived from various organic and inorganic acids and bases according to procedures known in the art. Pharmaceutically acceptable salt forms of the compounds of the invention are mostly prepared conventionally. If the compound according to the invention contains a carboxylic acid group, one of its suitable salts can be formed by reacting the compound with a suitable base to give the corresponding base addition salt. Such bases include, for example, alkali metal hydroxides, including potassium hydroxide, sodium hydroxide and lithium hydroxide; Alkaline earth metal hydroxides such as barium hydroxide and calcium hydroxide; Alkali metal alcoholates, eg, potassium ethanolate and sodium propanolate; and various organic bases such as piperidine, diethanolamine and N-methylglutamine. The aluminum salts of the compounds of formula I are also included. In certain compounds according to the invention, acid addition salts can be formed by reacting these compounds with pharmaceutically acceptable organic and inorganic acids, for example hydrogen halides such as hydrogen chloride, hydrogen bromide or hydrogen iodide, other mineral acids and their corresponding salts such as sulfate, nitrate or phosphate and the like, and alkyl and monoaryl sulfonates such as ethanesulfonate, toluenesulfonate and benzenesulfonate, as well as other organic acids and their corresponding salts such as acetate, trifluoroacetate, tartrate, maleate, succinate, citrate, benzoate, salicylate, ascorbate and the like. Accordingly, pharmaceutically acceptable acid addition salts of the compounds of formula I include the following: acetate, adipate, alginate, arginate, aspartate, benzoate, benzene sulfonate (besylate), bisulfate, bisulfite, bromide, butyrate, camphorate, camphorsulfonate, caprylate, chloride, chlorobenzoate, citrate, cyclopentanepropionate, digluconate, dihydrogenphosphate, dinitrobenzoate, dodecylsulfate, ethanesulfonate, fumarate, galacterate (from mucic acid), galacturonate, glucoheptanoate, gluconate , Glutamate, glycerophosphate, hemisuccinate, hemisulfate, heptanoate, hexanoate, hippurate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isethionate, isobutyrate, lactate, lactobionate, malate, maleate, malonate, mandelate, metaphosphate, methanesulfonate, methylbenzoate, monohydrogenphosphate , 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, oleate, pamoate, pectinate, persulfate, phenylacetate, 3-phenylpropionate, phosphate, phosphonate, phthalate, but this is not limiting.

Farther counting to the base salts of the compounds of the invention aluminum, Ammonium, calcium, copper, iron (III), iron (II), lithium, Magnesium, manganese (III), manganese (II), potassium, sodium and zinc salts, but no restriction should represent. Preferred among the above-mentioned salts Ammonium; the alkali metal salts sodium and potassium, and the alkaline earth metal salts Calcium and magnesium. To salts of the compounds according to the invention, derived from pharmaceutically acceptable non-toxic organic Derive bases, count Salts primary, secondary and tertiary Amines, substituted amines, including naturally occurring substituted Amines, cyclic amines and basic ion exchange resins, e.g. Arginine, betaine, caffeine, chloroprocaine, choline, N, N'-dibenzylethylenediamine (Benzathine), dicyclohexylamine, diethanolamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, iso-propylamine, Lidocaine, lysine, meglumine, N-methyl-D-glucamine, morpholine, piperazine, Piperidine, polyamine resins, procaine, purines, theobromine, triethanolamine, Triethylamine, trimethylamine, tripropylamine and tris (hydroxymethyl) methylamine (Tromethamine), which is not intended to be limiting.

Compounds of the present invention containing basic nitrogen-containing groups can be reacted with agents such as (C ₁ -C ₄ ) alkyl halides, eg, methyl, ethyl, isopropyl, and tert-butyl chloride, bromide, and iodide; Di (C ₁ -C ₄ ) alkyl sulfates, for example dimethyl, diethyl and diamyl sulfate; (C ₁₀ -C ₁₈ ) alkyl halides, for example decyl, dodecyl, lauryl, myristyl and stearyl chloride, bromide and iodide; and aryl (C ₁ -C ₄ ) alkyl halides, eg benzyl chloride and phenethyl bromide, quaternize. With such salts, both water- and oil-soluble compounds of the invention can be prepared.

To the abovementioned pharmaceutical salts, which are preferred counting Acetate, trifluoroacetate, besylate, citrate, fumarate, gluconate, hemisuccinate, Hippurate, hydrochloride, hydrobromide, isethionate, mandelate, meglumine, Nitrate, oleate, phosphonate, pivalate, sodium phosphate, stearate, sulphate, Sulfosalicylate, tartrate, thiomalate, tosylate and tromethamine, which but no limitation should represent.

The Acid addition salts basic compounds of the invention are made by that one the free base form with a sufficient amount of the desired Acid in Contact brings you on usual Way the salt represents. The free base can be brought by contacting the Salt form with a base and isolate the free base in the usual way regenerate. The free base forms differ in some way Sense of their corresponding salt forms in relation to certain physical properties such as solubility in polar solvents; in the context of the invention, however, the salts otherwise correspond to theirs respective free base forms.

As mentioned the pharmaceutically acceptable base addition salts of Compounds of the invention with metals or amines such as alkali metals and alkaline earth metals or organic amines. Preferred metals are sodium, Potassium, magnesium and calcium. Preferred organic amines are N, N'-dibenzylethylenediamine, Chloroprocaine, choline, diethanolamine, ethylenediamine, N-methyl-D-glucamine and procaine.

The Base addition salts of acidic compounds of the invention produced by the free acid form with a sufficient amount of the desired base in contact, causing the salt to usual Way. The free acid let yourself by contacting the salt form with an acid and Isolate the free acid on usual Regenerate way. The free acid forms differ In a sense, they are related to their corresponding salt forms to certain physical properties such as solubility in polar solvents; in the context of the invention, however, the salts otherwise correspond to theirs respective free acid forms.

If a compound according to the invention contains more than one group which can form such pharmaceutically acceptable salts, the invention also encompasses multiple salts. Typical multiple salt forms include, for example, bitartrate, diacetate, difumarate, dimeglumine, diphosphate, disodium and trihydro chloride, which is not intended to be limiting.

in the In view of the above, it can be seen that the term "pharmaceutically acceptable Salt "in the present Connection is to be understood as an active ingredient which is a compound of the invention in the form of one of its salts, especially if this salt form the active ingredient compared to the free form of Active ingredient or any other salt form of the active substance, the earlier used, imparts improved pharmacokinetic properties. The pharmaceutically acceptable salt form of the active ingredient may also this drug only a desired confer pharmacokinetic properties on which he did not previously has and even the pharmacodynamics of this drug to positively influence its therapeutic efficacy in the body.

Compounds of the invention can due to their molecular structure be and can be chiral Accordingly, they occur in different enantiomeric forms. You can therefore in racemic or optically active form.

There the pharmaceutical activity of racemates or stereoisomers the compounds of the invention may differ, it may be desirable be to use the enantiomers. In these cases, the end product or but already the intermediates in enantiomeric compounds, by chemical or physical measures known to the person skilled in the art, separated or already used as such in the synthesis become.

in the Fall racemic amines are from the mixture by reaction with formed an optically active release agent diastereomers. As a release agent are suitable e.g. optically active acids, such as the R and S forms of tartaric acid, diacetyl tartaric dibenzoyltartaric, Mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline) or the various optically active camphorsulfonic acids. Also advantageous is a chromatographic enantiomer separation with Aid of an optically active release agent (e.g., dinitrobenzoylphenylglycine, Cellulose triacetate or other derivatives of carbohydrates or on silica gel fixed chirally derivatized methacrylate polymers). As eluents are suitable for this purpose aqueous or alcoholic solvent mixtures such as. Hexane / isopropanol / acetonitrile e.g. in the ratio 82: 15: 3.

object The invention further relates to the use of the compounds and / or their physiologically acceptable salts for the preparation of a medicament (pharmaceutical preparation), in particular by non-chemical means. Here you can together with at least one solid, liquid and / or semi-liquid carrier or Excipient and optionally in combination with one or more further active ingredients are brought into a suitable dosage form.

object The invention furthermore relates to medicaments containing at least one inventive compound and / or their pharmaceutically usable derivatives, solvates and Stereoisomers, including theirs Mixtures in all proportions, and, where appropriate, carrier and / or auxiliaries.

pharmaceutical Formulations can in the form of dosage units containing a predetermined amount of active ingredient per dosage unit are included. Such a unit For example, 0.1 mg to 3 g, preferably 1 mg to 700 mg, particularly preferably 5 mg to 100 mg of a compound according to the invention depending on the disease condition being treated, the route of administration and the age, weight and condition of the patient, or pharmaceutical Formulations can in the form of dosage units containing a predetermined amount of active ingredient per dosage unit are included. Preferred dosage unit formulations are those that give a daily or partial dose, as stated above, or a corresponding fraction of an active ingredient. Furthermore, such pharmaceutical formulations can be included one of the methods well known in the pharmaceutical art produce.

pharmaceutical Formulations may be administered by any suitable route, for example, oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) routes. Such formulations can with all methods known in the pharmaceutical art by, for example, adding the active substance to the carrier (s) or adjuvant (s) is brought together.

Pharmaceutical formulations adapted for oral administration may be presented as separate entities, such as capsules or tablets; Powder or granules; Solutions or suspensions in aqueous or non-aqueous liquids; edible foams or foam foods; or oil-in-water liquid emulsions or water-in-oil liquid emulsions.

So let yourself for example, in oral administration in the form of a tablet or capsule the drug component with an oral, non-toxic and pharmaceutically acceptable inert carrier, such as e.g. ethanol, Glycerine, water and the like combine. Powders are made by adding the compound crushed a suitable fine size and with a similar one Way crushed pharmaceutical carrier, such. an edible carbohydrate such as starch or mannitol is mixed. A flavoring, preservative, Dispersant and dye may also be present.

capsules are prepared by mixing a powder mixture as described above prepared and shaped gelatin shells are filled with it. Lubricants such as e.g. fumed silica, talc, Magnesium stearate, calcium stearate or polyethylene glycol in solid form can the powder mixture before the filling process be added. A disintegrants or solubilizers, e.g. Agar Agar, Calcium carbonate or sodium carbonate, may also be added for availability improve the drug after taking the capsule.

In addition, if required or necessary, suitable binding, lubricating and disintegrating agents as well Dyes are also incorporated into the mixture. To the suitable binders include starch, gelatin, natural Sugars, e.g. Glucose or beta-lactose, sweeteners from corn, natural and synthetic Gum, e.g. Acacia, tragacanth or sodium alginate, carboxymethylcellulose, Polyethylene glycol, waxes, etc. The lubricants used in these dosage forms include sodium oleate, Sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, Sodium chloride and the like Belonging to the explosives without limitation to be, strength, Methyl cellulose, agar, bentonite, xanthan gum and the like. The tablets are formulated for example, by preparing a powder mixture, granulated or is pressed dry, a lubricant and a disintegrant are added and the Whole compressed into tablets becomes. A powder mixture is prepared by the in suitable Way crushed compound with a diluent or a base, as described above, and optionally with a binder, such as. Carboxymethylcellulose, an alginate, gelatin or polyvinylpyrrolidone, a solution slower, such as. Paraffin, a resorption accelerator, such as one quaternary Salt and / or an absorbent, e.g. Bentonite, kaolin or dicalcium phosphate. The powder mixture can be granulated by mixing with a binder, e.g. Syrup, starch paste, Acadia mucus or solutions made of cellulose or polymer materials wetted and through a sieve pressed becomes. As an alternative to granulation, the powder mixture can be used run through a tabletting machine, wherein unevenly shaped Lumps are formed, which are broken up into granules. The granules can by adding stearic acid, a stearate salt, talc or mineral oil are greased to stick at the tablet casting molds to prevent. The greased mixture is then compressed into tablets. The Compounds of the invention can also with a free-flowing inert carrier combined and then without implementation the granulation or dry compression steps directly pressed into tablets become. A transparent or opaque protective layer, consisting from a shellac sealant, a layer of sugar or Polymer material and a gloss layer of wax, may be present be. These coatings can Dyes are added to between different dosage units to be able to distinguish.

oral Liquids, such as. Solution, Syrups and elixirs, can be prepared in the form of dosage units, so that a given quantity contains a predetermined amount of the compound. Syrups can be prepared, by the compound in an aqueous solution with appropriate taste is solved, while Elixir using a non-toxic alcoholic vehicle getting produced. Suspensions can be obtained by dispersion of the compound be formulated in a non-toxic vehicle. solubilizers and emulsifying agents, e.g. ethoxylated isostearyl alcohols and Polyoxyethylene sorbitol ethers, preservatives, flavoring additives, such as e.g. peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, etc. can also be added.

The Dosage unit formulations for oral administration may optionally enclosed in microcapsules. The formulation can be also produce so that the Release prolonged or is retarded, such as by coating or embedding of particulate Material in polymers, wax, etc.

The compounds according to the invention as well as salts, solvates and physiologically functional derivatives thereof can also be administered in the form of liposome delivery systems, such as, for example, small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be different Phospholipids such as cholesterol, stearylamine or phosphatidylcholines.

The Compounds of the invention and the salts, solvates and physiologically functional derivatives of it can also using monoclonal antibodies as individual carriers the the connecting molecules coupled, fed become. The connections can also with soluble Polymers are coupled as targeted drug carrier. Such Polymers can Polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidophenol, Polyhydroxyethylaspartamidephenol or polyethyleneoxidepolylysine, substituted with palmitoyl radicals. Furthermore, the Compounds to a class of biodegradable polymers, for the controlled release of a drug are suitable, e.g. polylactic acid, Polyepsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, Polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels, coupled.

At the transdermal administration adapted pharmaceutical formulations can as independent Plaster for longer, close contact with the epidermis of the recipient be presented. So For example, the drug from the patch by iontophoresis supplied as described in Pharmaceutical Research, 3 (6), 318 (1986) described.

At the topical administration adapted pharmaceutical compounds can as ointments, creams, suspensions, lotions, powders, solutions, Be formulated pastes, gels, sprays, aerosols or oils.

For treatments of the eye or other external tissue, e.g. Mouth and skin, the formulations are preferably as topical Ointment or cream applied. When formulated into an ointment can the active ingredient either with a paraffinic or one with water Miscible cream base can be used. Alternatively, the active ingredient to a cream with an oil-in-water cream base or a water-in-oil basis be formulated.

To the adapted to the topical application to the eye pharmaceutical formulations include eye drops, wherein the active ingredient in a suitable carrier, in particular an aqueous solvent, dissolved or is suspended.

At the topical application in the mouth adapted pharmaceutical formulations include lozenges, lozenges and mouthwashes.

At rectal administration adapted pharmaceutical formulations can in the form of suppositories or enemas be presented.

At adapted the nasal administration pharmaceutical formulations in which the vehicle is a solid, contain a coarse powder with a particle size, for example in the range from 20-500 Micrometre, recorded in the manner of snuff is administered, i. by rapid inhalation over the Nasal passages from a container held close to the nose the powder. Suitable formulations for administration as a nasal spray or nose drops with a liquid as a carrier include drug solutions in water or oil.

At administration by inhalation adapted pharmaceutical formulations include fine particulate dusts or nebulae, which are pressurized by means of various types Dosing dispensers with aerosols, nebulizers or insufflators produced can be.

At the vaginal administration adapted pharmaceutical formulations can as pessaries, tampons, creams, gels, pastes, foams or spray formulations be presented.

To the pharmaceutical formulations adapted for parenteral administration belong to watery and non-aqueous sterile Injection solutions, the antioxidants, buffers, bacteriostats and solutes, by the the formulation is made isotonic with the blood of the recipient to be treated will contain; as well as aqueous and non-aqueous sterile Suspensions which may contain suspending agents and thickeners. The Formulations can in single or multiple dose containers, e.g. sealed ampoules and vials, presented and in freeze-dried (lyophilized) state be stored so that only the addition of the sterile carrier liquid, e.g. Water for Injections, needed immediately before use. Prescribed injection solutions and suspensions can from sterile powders, granules and tablets.

It understands that the Formulations in addition to the above-mentioned components especially others usual in the field Means with respect to the respective type of formulation included can; so can for example the oral administration suitable formulations flavorings contain.

A therapeutically effective amount of a compound of the invention depends on a number of factors, including e.g. the age and weight of the human being or animal, the exact state of the disease, which the Treatment needs, as well as its severity, the condition the formulation as well as the route of administration, and ultimately will determined by the attending physician or veterinarian. However, it lies an effective amount of a compound of the invention for the treatment generally in the range of 0.1 to 100 mg / kg of body weight Recipient (Mammal) per day and more typically in the range of 1 to 10 mg / kg of body weight per day. Thus would be for one 70 kg adult mammal the actual Amount per day for usually between 70 and 700 mg, taking this amount as a single dose per day or more usual in a series of sub-doses (such as two, three, four, five, or six) can be given per day, so that the total daily dose the same is. An effective amount of a salt or solvate or a physiologically functional derivative thereof may be used as a proportion the effective amount of the compound of the invention determined per se become. It can be assume that similar Dosages for the treatment of the other, above-mentioned disease states suitable are.

object The invention further comprises medicaments containing at least one inventive compound and / or their pharmaceutically usable derivatives, solvates and Stereoisomers, including theirs Mixtures in all proportions, and at least one other drug.

When other active pharmaceutical ingredients are preferred chemotherapeutic agents, especially those that inhibit angiogenesis and thereby growth and inhibit the proliferation of tumor cells; preferred are VEGF receptor inhibitors, including robozymes and antisense, which directed to VEGF receptors, as well as angiostatin and endostatin.

Examples antineoplastic agents used in combination with the compounds of the invention can be generally include alkylating agents, antimetabolites; Epidophyllotoxin; an antineoplastic enzyme; a topoisomerase inhibitor; procarbazine; Mitoxantrone or platinum coordination complexes.

Antineoplastic agents are preferably selected from the following classes:
Anthracyclines, vinca drugs, mitomycins, bleomycins, cytotoxic nucleosides, epothilones, discodermolides, pteridines, diynes and podophyllotoxins.

Especially preferred in the mentioned classes are e.g. Carminomycin, daunorubicin, Aminopterin, methotrexate, methopterine, dichloromethotrexate, mitomycin C, porfiromycin, 5-fluorouracil, 5-fluorodeoxyuridine monophosphate, Cytarabine, 5-azacytidine, thioguanine, azathioprine, adenosine, pentostatin, Erythrohydroxynonyladenine, cladribine, 6-mercaptopurine, gemcitabine, cytosine arabinoside, Podophyllotoxin or podophyllotoxin derivatives, e.g. Etoposide, Etoposide Phosphate or Teniposide, Melphalan, Vinblastine, Vinorelbine, Vincristine, Leurosidines, Vindesine, Leurosine, Docetaxel and Paclitaxel. Other preferred antineoplastic agents are selected from the group Discodermolide, Epothilone D, Estramustine, Carboplatin, Cisplatin, oxaliplatin, cyclophosphamide, bleomycin, gemcitabine, Ifosamide, melphalan, hexamethylmelamine, thiotepa, idatrexate, trimetrexate, Dacarbazine, L-asparaginase, camptothecin, CPT-11, topotecan, arabinosyl-cytosine, Bicalutamide, flutamide, leuprolide, pyridobenzoindole derivatives, interferons and interleukins.

As further active pharmaceutical ingredients antibiotics are preferred. Preferred antibiotics are selected from the group
Dactinomycin, daunorubicin, idarubicin, epirubicin, mitoxantrone, bleomycin, plicamycin, mitomycin.

When other active pharmaceutical ingredients are enzyme inhibitors preferred. Preferred enzyme inhibitors are selected from the group of histone deacetylation inhibitors (e.g., suberoylanilide hydroxamic acid [SAHA]) and tyrosine kinase inhibitors (e.g., ZD 1839 [Iressa]).

When other active pharmaceutical ingredients are nuclear export inhibitors preferred. Nuclear export inhibitors prevent the discharge of biopolymers (e.g., RNA) from the nucleus. Preferred nuclear export inhibitors are selected from the group Callystatin, Leptomycin B, Ratjadone.

As further active pharmaceutical ingredients, nuclear export inhibitors are preferred. Nuclear export Inhibito prevent the removal of biopolymers (eg RNA) from the cell nucleus. Preferred nuclear export inhibitors are selected from the group Callystatin, Leptomycin B, Ratjadone.

When other active pharmaceutical ingredients are immunosuppressants preferred. Preferred immunosuppressants are selected from the group rapamycin, CCI-779 (Wyeth), RAD001 (Novartis), AP23573 (Ariad Pharmaceuticals).

• (a) einer wirksamen Menge an einer erfindungsgemäßen Verbindung und/oder ihrer pharmazeutisch verwendbaren Derivate, Solvate und Stereoisomere, einschließlich deren Mischungen in allen Verhältnissen, und
• (b) einer wirksamen Menge eines weiteren Arzneimittelwirkstoffs.

The invention is also a set (kit), consisting of separate packages of

• (A) an effective amount of a compound of the invention and / or its pharmaceutically acceptable derivatives, solvates and stereoisomers, including mixtures thereof in all proportions, and
• (b) an effective amount of another drug.

The kit contains suitable containers, such as boxes or boxes, individual bottles, bags or ampoules. For example, the kit may contain separate ampoules each containing an effective amount of a compound of the invention and / or its pharmaceutically acceptable derivatives, solvates and stereoisomers, including mixtures thereof in all proportions,
and an effective amount of another drug ingredient is dissolved or in lyophilized form.

USE

The Present compounds are useful as pharmaceutical agents for mammals, especially for humans, in the treatment of diseases in which HSP90 plays a role.

object The invention thus relates to the use of compounds according to the invention, and their pharmaceutically usable derivatives, solvates and stereoisomers, including their mixtures in all proportions, for the manufacture of a medicament for the treatment of diseases, in which the inhibition, regulation and / or modulation of HSP90 plays a role.

The present invention comprises the use of the compounds according to the invention and / or their physiologically acceptable salts and solvates for the preparation of a medicament for the treatment of tumorous diseases, such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, Leiosarkom, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat carcinoma, sebaceous gland carcinoma, papillary carcinoma, Papillaradenokarzinomen, Cystadenokarzinomen, bone marrow carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma , Seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, epend ymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, leukemia, lymphoma, multiple myeloma, Waldenstrom's macroglobulinemia and severe chain disease; viral diseases, wherein the viral pathogen is selected from the group consisting of hepatitis type A, type B hepatitis, type C hepatitis, influenza, varicella, adenovirus, herpes simplex type I (HSV-I), herpes simplex type II (HSV -II), rinderpest, rhinovirus, echovirus, rotavirus, respiratory syncytial virus (RSV), papillomavirus, papovavirus, cytomegalovirus, echinovirus, arbovirus, huntavirus, coxsackievirus, mumps virus, measles virus, rubella virus, poliovirus, human immunodeficiency virus type I (HIV-I) and human immunodeficiency virus type II (HIV-II);
for immunosuppression in transplantations; inflammatory diseases such as rheumatoid arthritis, asthma, multiple sclerosis, type 1 diabetes, lupus erythematosus, psoriasis and inflammatory bowel disease; Cystic fibrosis; Diseases associated with angiogenesis such as diabetic retinopathy, hemangiomas, endometriosis, tumor angiogenesis; infectious diseases; Autoimmune diseases; ischemia; Promotion of nerve regeneration; fibrogenetic disorders such as scleroderma, polymyositis, systemic lupus, liver cirrhosis, keloid formation, interstitial nephritis and pulmonary fibrosis.

The Compounds of the invention can especially the growth of cancer, tumor cells and tumor metastases inhibit and are therefore for the tumor therapy suitable.

The present invention further comprises the use of the compounds according to the invention and / or their physiologically acceptable salts and solvates for the preparation of a medicament for the protection of normal cells against toxicity caused by chemotherapy and for the treatment of diseases protein misfolding or aggregation is a major causative factor such as scrapie, Creutzfeldt-Jakob disease, Huntington's or Alzheimer's.

The The invention also relates to the use of the compounds according to the invention and / or their physiologically acceptable salts and solvates Preparation of a medicament for the treatment of diseases of the Central nervous system, cardiovascular disease and cachexia.

The The invention also relates to the use in a further embodiment the compounds of the invention and / or their physiologically acceptable salts and solvates Preparation of a medicament for HSP90 modulation, wherein the modulated HSP90 biological activity an immune response in one Individual, protein transport from endoplasmic reticulum, recovery from hypoxic / anoxic stress, recovery from malnutrition, recovery of heat stress, or combinations thereof, and / or being the disorder a type of cancer is an infectious disease, a disorder that with a disturbed Protein transport from the endoplasmic reticulum, a disorder that with ischemia / reperfusion or combinations thereof, those with ischemia / reperfusion accompanying disorder a consequence of cardiac arrest, asystole and delayed ventricular arrhythmias, Cardiac surgery, cardiopulmonary bypass surgery, Organ transplantation, spinal cord injury, Head trauma, stroke, thromboembolic stroke, hemorrhagic Stroke, cerebral vasospasm, hypotension, hypoglycemia, status epileptic seizure, anxiety, schizophrenia, one neurodegenerative disorder, Alzheimer's disease, Huntington's chorea, amyotrophic lateral sclerosis (ALS) or stress in the newborn.

The The invention also relates to the use in a further embodiment the compounds of the invention and / or their physiologically acceptable salts and solvates Preparation of a medicament for treating ischemia Consequence of cardiac arrest, asystole and delayed ventricular arrhythmias, Heart surgery, cardiopulmonary Bypass surgery, organ transplantation, spinal cord injury, head trauma, Stroke, thromboembolic stroke, haemorrhagic stroke, cerebral vasospasm, hypotension, hypoglycemia, status epilepticus, one epileptic seizure, anxiety, schizophrenia, a neurodegenerative disorder Alzheimer's disease, Huntington's chorea, amyotrophic lateral sclerosis (ALS) or stress in the newborn.

Test method for measurement of HSP90 inhibitors

The Binding of geldanamycin or 17-allylamino-17-demethoxygeldanamycin (17AAG) and its competitive inhibition on HSP90 can be used about the inhibitory activity the compounds of the invention (Carreras et al., 2003, Chiosis et al., 2002). Particularly Case a radioligand filter binding test is used. As radioligand thereby tritiated 17-allylamino-geldanamycin, [3H] 17AAG, used. This filter binding test allows a targeted search after inhibitors that interfere with the ATP binding site.

material

• Recombinant human HSP90α (expressing E. coli, 95% purity);
• [3H] 17AAG (17-allylamino-geldanamycin, [allylamino-2,3- ³ H. Specific
• Activity: 1.11 x 10 ¹² Bq / mmol (Moravek, MT-1717);
• HEPES filter buffer (50 mM HEPES, pH 7.0, 5 mM MgCl 2, BSA 0.01%) Multiscreen FB (1 μm) Filter plate (Millipore, MAFBNOB 50).

method

The 96 well microtiter filter plates are first watered and with 0.1% polyethyleneimine coated.

• Reaktionstemperatur 22°C
• Reaktionszeit: 30 min., Schütteln bei 800 upm
• Testvolumen: 50 μl
• Endkonzentrationen:
• 50 mM HEPES-HCl, pH 7,0, 5 mM MgCl2, 0,01% (w/v) BSA
• HSP90: 1,5 μg/assay
• [3H]17AAG: 0,08 μM.

The test is carried out under the following conditions:

• Reaction temperature 22 ° C
• Reaction time: 30 min., Shaking at 800 rpm
• Test volume: 50 μl
• final concentrations:
• 50 mM HEPES-HCl, pH 7.0, 5 mM MgCl 2, 0.01% (w / v) BSA
• HSP90: 1.5 μg / assay
• [3H] 17AAG: 0.08 μM.

At the End of the reaction becomes the supernatant in the filter plate with the help of a vacuum manifold (Multiscreen Separation System, Millipore) and the filter washed twice.

The Filter plates are then placed in a beta counter (Microbeta, Wallac) measured with scintillator (Microscint 20, Packard).

Out the "counts per minutes "values becomes "% the control " and from this the IC-50 value of a connection is calculated.

In front- and below, all temperatures are in ° C. In the following Examples means "usual workup": one gives, if required to add water, if necessary, depending on Constitution of the final product to pH values between 2 and 10, extracted with ethyl acetate or dichloromethane, separated, dried the organic phase over Sodium sulfate, evaporated and purified by chromatography on silica gel and / or by crystallization. Rf values on silica gel; Eluent: Ethyl acetate / methanol 9: 1.

The retention times RT [min] and M + H ⁺ data MW given in the examples below are the measurement results of the HPLC and the LC-MS measurements.

HPLC conditions

HPLC method ^a)

• Gradient: 5.5 min; Flow .: 2.75 ml / min from 90:10 to - 0: 100 H ₂ O / ACN water + TFA (0.01% vol.); Acetonitrile + TFA (0.01% by volume)
• Pillar: Chromolith SpeedROD RP 18e 50-4.6
• Wavelength: 220 nm

HPLC method ^b)

• Gradient: 5.5 min; Flow: 2.75 ml / min from 99: 1 to - 0: 100 H ₂ O / ACN water + TFA (0.01% vol.); Acetonitrile + TFA (0.01% by volume)
• Pillar: Chromolith SpeedROD RP 18e 50-4.6
• Wavelength: 220 nm

LC-MS conditions

• Hewlett Packard HP 1100 series system with the following features:
• Ion source: electrospray (positive mode); Scan: 100-1000 m / z;
• Fragmentation voltage: 60 V; Gas temperature: 300 ° C, DAD: 220 nm.
• Flow rate: 2.4 ml / min. The splitter used reduced after the DAD the flow rate for the MS to 0.75 ml / min.
• Pillar: Chromolith SpeedROD RP-18e 50-4.6
• Solvent: LiChrosolv Quality the company Merck KGaA

gradient: P1: Solvent A: H2O (0.1% TFA) Solvent B: ACN (0.1% TFA) 99% A → 100% B: running time 6 minutes / flow rate 3.0 ml / minute P2: Solvent A: H2O (0.01% TFA) Solvent B: ACN (0.01% TFA) 99% A → 100% B: run time 5 minutes / flow rate 2.75 ml / minute N: Solvent A: H2O (0.01% TFA) Solvent B: ACN (0.01% TFA) 90% A → 100% B: run time 5 minutes / flow rate 2.75 ml / minute

example 1

Preparation of 2-amino-4-chloro-6-phenyl-pyrimidine ( "A1")

To 250 mg of 2-amino-4,6-dichloropyrimidine and 186 mg of benzoic boronic acid are added 12.5 ml of ethylene glycol dimethyl ether. Then a solution of 0.5 g of Na ₂ CO ₃ in a little water is added, followed by 17 mg of palladium (II) acetate and 39 mg of triphenylphosphine. The mixture is refluxed for 12 hours. It is cooled, the solvent is removed and worked up as usual. The crude product is chromatographed on a 120 g Redisep silica gel column of a Combiflashanlage. This gives 186 mg of "A1"; RT 2.25, HPLC method ^a) ; [M + H] ⁺ 206.

Analogously, the following compounds are obtained

Example 2

• 2.1 3 ml 4-Methylbenzaldehyd und 2,33 ml Methylcyanacetat werden unter Kühlung mit 7,5 g Aluminiumoxid (90 aktiv neutral) versetzt und 10 Minuten bei Raumtemperatur gerührt. Das Gemisch wird in Ethylacetat verrührt. Man filtriert und erhält nach weiterer Aufarbeitung 4,15 g 2-Cyan-3-p-tolyl-acrylsäure-methylester ("2a").
• 2.2 Zu einer Suspension von 1,0 g "2a" und 0,475 g Guanidiniumchlorid in 10 ml 1-Butanol gibt man eine Lösung von 0,114 Natrium in 0,29 ml Ethanol. Man rührt 4 Stunden bei 100°. Man kühlt ab, gießt auf Eis und neutralisiert mit Essigsäure. Der Feststoff wird abgesaugt und mit Wasser nachgewaschen. Das Kristallisat wird mit Ethylacetat und Methanol verrieben, abgesaugt und getrocknet. Man erhält 253 mg 2-Amino-4-hydroxy-5-cyan-6-(4-methyl-phenyl)-pyrimidin ("2b").
• 2.3 50 mg "2b" werden in 0,3 ml POCl₃ suspendiert und unter Argonatmosphäre 2 Stunden unter Rückfluß gerührt. Man gießt auf Eis, trennt den ausgefallenen Feststoff ab und reinigt mittels Combiflash-Chromatographie. Man erhält 9,8 mg "A14", RT 2.13 ^a); [M+H]⁺ 245.

The preparation of 2-amino-4-chloro-5-cyano-6- (4-methyl-phenyl) -pyrimidine ("A14") is carried out analogously to the following scheme:

• 2.1 3 ml of 4-methylbenzaldehyde and 2.33 ml of methyl cyanoacetate are added with cooling with 7.5 g of aluminum oxide (90 active neutral) and stirred for 10 minutes at room temperature. The mixture is stirred in ethyl acetate. It is filtered and receives after further work-up 4.15 g of methyl 2-cyano-3-p-tolyl-acrylate ("2a").
• 2.2 To a suspension of 1.0 g of "2a" and 0.475 g of guanidinium chloride in 10 ml of 1-butanol is added a solution of 0.114 sodium in 0.29 ml of ethanol. It is stirred for 4 hours at 100 °. It is cooled, poured on ice and neutralized with acetic acid. The solid is filtered off with suction and washed with water. The crystals are triturated with ethyl acetate and methanol, filtered off with suction and dried. This gives 253 mg of 2-amino-4-hydroxy-5-cyano-6- (4-methyl-phenyl) -pyrimidine ("2b").
• 2.3 50 mg "2b" are suspended in 0.3 ml POCl ₃ and stirred under argon atmosphere for 2 hours under reflux. It is poured onto ice, the precipitated solid is separated off and purified by means of Combiflash chromatography. This gives 9.8 mg of "A14", RT 2.13 ^a) ; [M + H] ⁺ 245.

Analogously, the following compounds are obtained

Pharmacological data

Affinity to receptors Table 1

• IC ₅₀ : 10 nM - 1 μM = A 1 μM - 10 μM = B> 10 μM = C

The The following examples relate to pharmaceutical preparations:

Example A: Injection glasses

A solution of 100 g of an active ingredient according to the invention and 5 g of disodium hydrogen phosphate is distilled twice in 3 l Water with 2 N hydrochloric acid adjusted to pH 6.5, sterile filtered, filled into injection jars, under sterile conditions and lyophilized sterile. each Contains injection glass 5 mg of active ingredient.

Example B: Suppositories

you melts a mixture of 20 g of an active ingredient according to the invention with 100 g soy lecithin and 1400 g cocoa butter, pour into molds and lets cool. Each suppository contains 20 mg of active ingredient.

Example C: Solution

A solution of 1 g of an active ingredient is prepared according to the invention, 9.38 g of NaH ₂ PO ₄ · 2 H ₂ O, 28.48 g Na ₂ HPO ₄ · 12 H ₂ O and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water , Adjust to pH 6.8, make up to 1 liter and sterilize by irradiation. This solution can be used in the form of eye drops.

Example D: Ointment

you mixes 500 mg of an active ingredient according to the invention with 99.5 g Vaseline under aseptic conditions.

Example E: Tablets

One Mixture of 1 kg of active ingredient, 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate is in the usual way to tablets pressed, such that each Tablet contains 10 mg of active ingredient.

Example F: dragees

Analogous Example E tablets are pressed, which are then in the usual Way with a plating Sucrose, potato starch, Talc, tragacanth and dyestuff coated become.

Example G: Capsules

2 kg of active ingredient are in common Way filled in hard gelatin capsules, so that everyone Capsule contains 20 mg of the active ingredient.

Example H: Ampoules

A solution of 1 kg of an active ingredient according to the invention in 60 l of double distilled water is filtered sterile, in Filled ampoules, lyophilized under sterile conditions and sealed sterile. Each ampoule contains 10 mg of active ingredient.

Claims (12)

Compounds selected from the group

and their pharmaceutically usable derivatives, solvates, salts, tautomers and stereoisomers, including mixtures thereof in all ratios. Medicament containing at least one compound according to claim 1 and / or its pharmaceutically usable derivatives, Salts, solvates, tautomers and stereoisomers, including theirs Mixtures in all proportions, and optionally excipients and / or adjuvants. Use of compounds according to claim 1 and their pharmaceutically usable derivatives, salts, solvates, tautomers and stereoisomers, including their mixtures in all proportions, for the manufacture of a medicament for the treatment and / or prophylaxis of diseases involving inhibition, regulation and / or modulation of HSP90 plays a role. Use according to claim 3 of compounds according to Claim 1 and its pharmaceutically usable derivatives, salts, Solvates, tautomers and stereoisomers, including mixtures thereof in all ratios, for the manufacture of a medicament for the treatment or prevention of tumors, viral diseases, for immunosuppression in transplants, inflammatory Diseases, Cystic fibrosis, related diseases Angiogenesis, infectious Diseases, autoimmune diseases, ischaemia, fibrogenetic diseases, to advancement nerve regeneration, to inhibit the growth of cancer, tumor cells and tumor metastases, to protect normal cells against toxicity caused by Chemotherapy is caused, for the treatment of diseases, where protein misfolding or aggregation is a major causative factor is. Use according to claim 4, wherein the Tumor diseases involving fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, Lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, Leiosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, Ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, Adenocarcinoma, sweat gland carcinoma, Sebaceous gland carcinoma, Papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, Bone marrow carcinoma, bronchogenic carcinoma, renal cell carcinoma, Hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonic Carcinoma, Wilms' tumor, Cervix cancer, Testicular tumor, Lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, Astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, Hemangioblastoma, acoustic Neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma, Leukemia, lymphoma, multiple myeloma, Waldenströms macroglobulinemia and severe chain disease. Use according to claim 4, wherein the viral pathogen the viral diseases selected is from the group consisting of hepatitis type A, hepatitis type B, hepatitis type C, influenza, varicella, adenovirus, herpes simplex Type I (HSV-I), Herpes Simplex Type II (HSV-II), Rinderpest, Rhinovirus, Echovirus, rotavirus, respiratory syncytial virus (RSV), papillomavirus, Papovavirus, cytomegalovirus, echinovirus, arbovirus, huntavirus, Coxsackie virus, mumps virus, measles virus, rubella virus, poliovirus, human immunodeficiency virus Type I (HIV-I) and Human Immunodeficiency Virus Type II (HIV-II). Use according to claim 4, wherein the inflammation-related Rheumatoid arthritis, asthma, multiple sclerosis, Type 1 Diabetes, Lupus Erythematosus, Psoriasis and Inflammatory Bowel Disease trades. Use according to claim 4, wherein the Diseases related to angiogenesis for diabetic retinopathy, hemangiomas, Endometriosis and tumor angiogenesis. Use according to claim 4, wherein the fibrogenetic disorders related to scleroderma, polymyositis, systemic Lupus, liver cirrhosis, keloid formation, interstitial nephritis and pulmonary fibrosis. Use according to claim 4, wherein the Diseases in which protein misfolding or aggregation is a major causal factor is to scrapie, Creutzfeldt-Jakob disease, Huntington's or Alzheimer's. Medicaments containing at least one compound according to claim 1 and / or its pharmaceutically usable derivatives, Salts, solvates, tautomers and stereoisomers, including theirs Mixtures in all proportions, and at least one other drug. A kit consisting of separate packages of (a) an effective amount of a compound of claim 1 and / or its pharmaceutically acceptable composition derivatives, salts, solvates, tautomers and stereoisomers, including mixtures thereof in all ratios, and (b) an effective amount of another drug active ingredient.