WO2001016124A1 - Cinnamic acid nitrile method of production and use thereof - Google Patents

Cinnamic acid nitrile method of production and use thereof Download PDF

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Publication number
WO2001016124A1
WO2001016124A1 PCT/EP2000/008419 EP0008419W WO0116124A1 WO 2001016124 A1 WO2001016124 A1 WO 2001016124A1 EP 0008419 W EP0008419 W EP 0008419W WO 0116124 A1 WO0116124 A1 WO 0116124A1
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alkyl
piperazinyl
alkoxyphenyl
general formula
alkoxy
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PCT/EP2000/008419
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German (de)
French (fr)
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Hermann Hauer
Egon Koch
Shyam Sunder Chatterjee
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Dr. Willmar Schwabe Gmbh & Co.
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Priority to AU70006/00A priority Critical patent/AU7000600A/en
Priority to EP00958512A priority patent/EP1208093A1/en
Publication of WO2001016124A1 publication Critical patent/WO2001016124A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • C07D295/182—Radicals derived from carboxylic acids
    • C07D295/185—Radicals derived from carboxylic acids from aliphatic carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/08—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms
    • C07D211/18—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D211/20—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by singly bound oxygen or sulphur atoms
    • C07D211/22—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by singly bound oxygen or sulphur atoms by oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no 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, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D211/60—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D211/62—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals attached in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D227/00—Heterocyclic compounds containing rings having one nitrogen atom as the only ring hetero atom, according to more than one of groups C07D203/00 - C07D225/00
    • C07D227/02—Heterocyclic compounds containing rings having one nitrogen atom as the only ring hetero atom, according to more than one of groups C07D203/00 - C07D225/00 with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D227/04—Heterocyclic compounds containing rings having one nitrogen atom as the only ring hetero atom, according to more than one of groups C07D203/00 - C07D225/00 with only hydrogen or carbon atoms directly attached to the ring nitrogen atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/08—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms
    • C07D295/084—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/088—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/12—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms
    • C07D295/125—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/13—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain

Definitions

  • Cinnamic acid nitriles process for their preparation and use
  • Benign prostatic hyperplasia is by far the most important urological disease in men.
  • BPH is a benign growth of epithelial and stromal parts of the prostate, with enlargement of the organ leading to urinary drainage disorders that are accompanied by symptoms such as pollakiuria and nocturia, as well as incomplete and delayed bladder emptying.
  • urinary congestion can lead to renal failure and uremia.
  • LUTS Lower Urinary Tract Symptoms
  • alpha antagonists and 5-alpha reductase inhibitors are primarily used as medicines for the treatment of BPH (see Eckert, RE, et al., Cellular basis of dynamic, infravesical obstruction in the context of a benign prostatic hyperplasia; role of alpha-receptor blockers and cyclic nucleotides, Akt. Urol. 29, 252-260 (1998); Weckermann, D. and Wawroschek, F. Medicinal therapy of benign prostatic hyperplasia syndrome. Münch. med. Wschr. 141, 54- 58 (1999); Weckermann, D. and Wawroschek, F. Interventional therapy for benign prostatic hyperplasia, Münch. Med. Wschr., 141, 59-63 (1999).
  • Alphar antagonists are based on the view that the obstruction of the urinary tract in BPH is based not only on a static component, which is caused by the enlarged prostate, but also on a dynamic component, which is caused by an increased tone of the smooth prostatic muscles.
  • a large number of studies have shown that dynamic obstruction in BPH is apparently mediated by an increased release of norepinephrine from sympathetic neurons. It is generally accepted today that in the Prostate predominantly alpha 1A adrenoceptors are expressed.
  • the clinical effectiveness of blocking alpha-adrenoceptors in the treatment of BPH was originally demonstrated using the non-specific antagonist phenoxybenzamine.
  • alpha blockers Because of the significant cardiovascular side effects, a number of so-called uroselective alpha- A blockers have been developed in recent years, which are much better tolerated. The results of clinical studies show that these substances have a statistically and clinically significant improvement in symptoms and maximum urine flow compared to placebo.
  • the advantage of alpha blockers is their rapid onset of action, but the growth potential of the prostate remains unaffected by these substances (cf. Heimbach, D. and Müller, SC. Treatment of BPH with ⁇ adrenoceptor antagonists. Urologe [A] 36, 18-34 (1997); Eckert et al., Loc. Cit .; Weckermann and Wawroschek, loc. Cit.).
  • BPH only develops in the presence of biologically active male sex hormones. BPH is practically never observed in men who had to undergo castration before the age of 40 or who have no or only insufficient androgen formation in the gonads due to an underactive pituitary gland. Likewise, in the event of a congenital defect or a lack of androgen receptors (e.g. testicular feminization), the normal development of the prostate and the formation of hyperplastic changes with increasing age do not occur. The most biologically important androgen in the prostate is dihydrotestosterone (DHT), which is produced locally under the influence of 5-alpha reductase from testosterone.
  • DHT dihydrotestosterone
  • DHT mainly stimulates the epithelial parts of BPH
  • inhibition of 5-alpha reductase leads to growth arrest or atrophy of the glandular part, but the stromal component of BPH is practically unaffected.
  • taking the 5-alpha reductase inhibitor finasteride results in a reduction in the prostatic DHT concentration of up to 85%, but the reduction in prostate size is only about 20% on average and also requires a period of up to 12 months , This effect is only clinically relevant if the prostate volume at the beginning of therapy is over 40 ml (see Geller, J. Pathogenesis and medical treatment of benign prostatic hyperplasia. Prostate (Suppl.) 2, 95-104 (1989 ); Weckermann and Wawroschek, loc.cit.).
  • epidermal growth factor [EGFJ, transforming growth factor- ⁇ , [TGF- ⁇ ], TGF-ß, basic fibroblast growth factor (bFGF), keratinocyte growth factor [KGF], nerve growth Factor [NGF], Insulin-like Growth Factor I [IGF-1] etc.) and their receptors.
  • BPH basic fibroblast growth factor
  • KGF keratinocyte growth factor
  • NGF nerve growth Factor
  • IGF-1 Insulin-like Growth Factor I
  • Prostate 28 392-405 (1996); Peehl, DM Celluiar biology of prostatic growth factors. Prostate (Suppl.) 6: 74-78 (1996); Peehl, DM and Seilers, RG Basic FGF, EGF, and PDGF modify TGFß-induction of smooth muscle cell phenotype in human prostatic stromal cells. Prostate 35, 125-134 (1998).
  • the invention is therefore based on the object of providing substances which, by inhibiting alphar adrenoceptors ( ⁇ i-antagonistic action) and by suppressing the growth factor-mediated cell proliferation (proliferation-inhibiting action) of epithelial and stromal cells, both the dynamic and the static Affect components of BPH positively and thus allow comprehensive treatment of BPH syndrome.
  • the object of the invention is also the provision of medicaments for the prophylaxis and treatment of BPH and of processes for their production and use.
  • X CN, COOR 4 , CONR 5 R 6 , COAryl, COalkyl, aryl, heteroaryl,
  • R 1 H, alkyl, arylalkyl, [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl,
  • R 2 and R 3 independently of one another H, hydroxy, alkoxy, benzyl,
  • R H, alkyl, [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl,
  • R 5 H, alkyl, alkoxyalkyl, alkoxy-oxoalkyl, dialkylaminoalkyl, amino-oxoalkyl, aryl,
  • alkyl in all cases is a straight-chain or branched alkyl radical having 1 to 8 carbon atoms, preferably having 1 to 4 carbon atoms
  • alkoxy is an alkoxy group having 1 to 4 carbon atoms Atoms, preferably with one or two carbon atoms.
  • Aryl is also to be understood as meaning any substituted aryl radicals, for example halophenyl, alkylphenyl, alkoxyphenyl,
  • the compounds according to the invention are substituted by radicals or groups which contain basic nitrogen atoms, for example cyclic amine groups such as pyrrolidine, morpholine, piperidine, piperazine, these compounds can be present either in the form of free bases or in the form of addition salts with pharmacologically acceptable acids, for example in Form of their hydrochloride or trifluoroacetate.
  • cyclic amine groups such as pyrrolidine, morpholine, piperidine, piperazine
  • Preferred cinnamic acid nitrile derivatives are those compounds of general forms I in which the at least one [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl radical is a [4- (2-methoxyphenyl) -1-piperazinyl] propyl radical is.
  • the compounds according to the invention are pharmacologically active; Their pharmacological effect is characterized by the simultaneous presence of an ⁇ T adrenoceptor-antagonistic active component and a proliferation-inhibiting active component.
  • the invention therefore also relates to medicaments which, as an active ingredient, contain, in addition to customary auxiliaries and additives, an amount of one or more of the compounds of general forms I according to the invention which is effective for the prophylaxis or treatment of BPH.
  • these medicaments can include, for example, water, vegetable oils, polyethylene glycols, glycerol esters, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, Vaseline®, preservatives, wetting agents, emulsifiers, physiologically acceptable salts, buffer substances, colorants, flavorings and flavorings contain.
  • the choice of excipients depends on the desired application form such as. B. tablets, dragees, juices, ampoules, suppositories, ointments or sprays.
  • the compounds according to the invention can also be administered in a mixture with other known active compounds.
  • the compound (E) -1 - [2-cyano-3- (3,4-dihydroxyphenyl) -1-oxo-2-propenyl] -4-phenylpiperazine is known from A. Gazit et al., J. Med Chem. 34, pp. 1896-1907 (1991) (there compound no. 68).
  • This known compound differs from the compounds of the general formula I according to the invention in that it contains a [4-phenyl-1-piperazinyl] radical, but not at least one [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl -Rest.
  • a proliferation-inhibiting effect is described for the known compound, it is not known whether this compound also has an ⁇ i-antagonistic effect.
  • the invention finally also relates to processes for the preparation of the compounds of the general formula I in which a benzaldehyde of the general formula II, in which R 1 , R 2 and R 3 have the meanings given above, with a substituted acetonitrile of the general formula III, in which X has the meaning given above, is implemented under conditions known for Knoevenagel condensations, ie aldol condensations.
  • the Knoevenagel condensation can be carried out with or without a catalyst.
  • Piperidine, ammonium acetate, ⁇ -alanine or glacial acetic acid serve as catalysts.
  • R 1 [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl
  • R 2 and R 3 independently of one another H, hydroxy, alkoxy with 1 - 4, preferably 1 - 2 C-
  • Atoms, benzyl are new reactive intermediates for the preparation of the compounds of general formula I, in which R 1 , R 2 and R 3 have the corresponding meanings. These intermediates are also the subject of the invention.
  • Process A The benzaldehyde of the general formula II correspondingly substituted with R 1 to R 3 , if appropriate released from its hydrochloride, 0.9 to 2.0 equivalents of the acetonitrile of the general formula III substituted with X and optionally a catalytic amount of piperidine are dissolved in ethanol, methanol , 2-propanol or TBME 2 h to 3 d at 50 ° C to boiling temperature.
  • the product is worked up by filtering off the product, by evaporation or by partitioning between ethyl acetate or chloroform and water. Purification is carried out by chromatography, recrystallization and / or salt formation using method C.
  • Process B The benzaldehyde of the general formula II substituted with R 1 to R 3 , if appropriate released from its hydrochloride, 0.9 to 1.1 equivalents of the Acetonitrile of the general formula III corresponding to X and optionally a catalytic amount of piperidine and / or glacial acetic acid are stirred in toluene for 2 to 24 hours on a water separator under reflux. The mixture is worked up by evaporating the reaction solution. Purification is carried out by chromatography, recrystallization and / or salt formation using method C.
  • Process C The base prepared by process A or B is dissolved in ethanol, 2-propanol or acetone and 1 to 3 equivalents of a solution of hydrogen chloride in 2-propanol or aqueous hydrochloric acid are added.
  • the hydrochloride is suctioned off or concentrated and purified by recrystallization.
  • Method E 2.0 to 2.4 equivalents of potassium hydroxide are added to 1.0 to 1.2 equivalents of the appropriately substituted 1- (3-chloropropyl) -4- (2-alkoxyphenyl) piperazine dihydrochloride in ethanol or 1-propanol.
  • potassium iodide 1.0 to 1.2 equivalents of potassium carbonate
  • the inorganic material is filtered off and the filtrate is spun in.
  • Working up is carried out by extraction with ethyl acetate or TBME and renewed filtration and / or by partitioning between ethyl acetate or TBME and water. It is cleaned by recrystallization or salt formation according to method C.
  • Process F 1.0 equivalent of isovanillin, 1.5 equivalents of 1-bromo-2-chloroethane or 1, 4-dibromobutane and 1.5 equivalents of potassium carbonate are stirred in 2-butanone at boiling temperature for 4 to 20 h. The inorganic material is filtered off and the filtrate is spun in. The purification is carried out by chromatography and / or recrystallization.
  • Process G The amine HNR 5 R 6 , optionally released from its hydrochloride, is reacted with 1.0 to 1.05 equivalents of methyl cyanoacetate in methanol, ethanol or without solvent for 15 min to 28 h at room temperature to 100 ° C. After cooling, the mixture is optionally rotated in and purified by recrystallization.
  • Method H The alcohol H ⁇ R 4 is heated with 5 equivalents of methyl cyanoacetate to 80 ° C. for 7 hours at a slightly reduced pressure. After cooling, it is purified by column chromatography and subsequent recrystallization.
  • a The pharmacological testing of the substances for interaction with alpha-adrenoceptors was carried out using a receptor binding assay using rat brain cell membranes.
  • male Sprague-Dawley rats 150-250 g were euthanized in CO 2 anesthesia and the brains (without the cerebellum) were removed. After removal of adhering blood and cerebral membranes, the brains were immediately removed in a 10-fold volume of ice-cold homogenization buffer (50 mM Tris-HCl, pH 7.4) and homogenized with an ice-cooled glass homogenizer.
  • the cell homogenate was centrifuged for 10 min at 50,000 g (4 ⁇ C) and the pellet resuspended in ice-cold homogenization buffer. After renewed centrifugation (10 min at 4 ° C. and 50,000 g), the cell membranes were in a 10-fold volume of binding buffer (50 mM Tris-HCl, 0.5 mM Na-EDTA, 0.01% ascorbic acid, 10 ⁇ M pargyline, pH 7, 4) and stored in portions (1 ml) at -80 ° C
  • the substances were dissolved in 150 ⁇ l binding buffer using DMSO and together with 50 ⁇ l brain cell membranes (2.5 mg / ml protein) and 50 ⁇ l 3 H-Prazos ⁇ n (300 pM, specific activity 80 Ci / mmol) in Binding buffer incubated for 45 min at room temperature. The non-specific binding was determined in the presence of 2 ⁇ M phentolamine.
  • the reaction mixture was then filtered through glass fiber filters (type GF / B) which had been pretreated overnight with polyethyleneimine (0.2% in distilled water). After washing four times with 3 ml of ice-cold binding buffer, the filters were dried at 60 ° C. for 1 h.
  • the radioactivity bound was determined after transferring the filter to 4 ml of scintillation liquid (Quickszint) in a beta counter.
  • the inhibition of the specific binding of 3 H-Prazos ⁇ n to Alpha 1 adenoceptors was calculated in percent of a solvent control examined in parallel
  • DMEM modified Eagle's Medium
  • FCS fetal calf serum
  • FCS fetal calf serum
  • FCS fetal calf serum
  • the cells were transferred to F-bottom microtiter plates in a volume of 200 ⁇ l per well and incubated for 96 h Medium (DMEM without FCS) and the substances were added 60 minutes later, the cell proliferation was restimulated by adding 10 ng / ml of recombinant human platelet-derived growth factor-BB (PDGF-BB). The cells were then again at 37 ° for 24 h C cultivated in the incubator Six hours before the cell harvest, 0.5 ⁇ Ci methyl 3 H-thyme was added per test batch after the 24-hour inku had expired During the bation period, the microtiter plates were centrifuged for 5 min at 400 g and the cell supernatants were carefully pipetted off.
  • DMEM fetal-derived growth factor-BB
  • the cells were detached from the ground with trypsin / EDTA and then harvested with a cell harvester (Inotech) on glass fiber filter (type G-10, ICH-201).
  • a cell harvester Inotech
  • the incorporation of 3 H-thymid into newly synthesized DNA was carried out with the aid of a linear analyzer (LB 2842, Berthold).
  • the degree of the anti-proactive effect of the substances was determined in each case in comparison to the solvent controls examined at the same time
  • the active ingredient is optionally ground, homogeneously mixed with the excipients and compressed in the usual manner to tablets each weighing 250 mg and 9 mm in diameter. If desired, the tablets are coated with a film.
  • the finely powdered substances are mixed homogeneously and filled into size 2 hard gelatin capsules in an amount of 200 mg per capsule.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Hydrogenated Pyridines (AREA)

Abstract

The invention relates to substituted cinnamic acid nitriles of the formula (I) wherein X = CN, COOR?4, CONR5R6¿, COaryl, COalkyl, aryl, heteroaryl, R1 = H, alkyl, arylalkyl, [4-(2-alkoxyphenyl)-1-piperazinyl]alkyl, R?2 and R3¿ = independent of each other H, hydroxy, alkoxy, benzyl, R4 = H, alkyl, [4-(2-alkoxyphenyl)-1-piperazinyl]alkyl, R5 = H, alkyl, alkoxyalkyl, alkoxy-oxoalkyl, diakylaminoalkyl, amino-oxoalkyl, aryl, arylalkyl, [4-(2-alkoxyphenyl)-1-piperazinyl]alkyl, morpholinoalkyl, furanyalkyl, thienylalkyl, R6 = H, alkyl, or NR5R6 together = cyclic amines such as pyrrolidine, morpholine, piperidine, subst. piperazine, whereby at least one of the radicals R?1, R4 or R5¿ is a [4-(2-alkoxyphenyl)-1-piperazinyl]alkyl-radical, in addition to the addition thereof with pharmacologically compatible acids. The compounds are active as prophylactic and therapeutic medicaments for the treatment of benign prostatahyperplasia. A method for the production of cinnamic acid nitrile is also disclosed; an appropriately substituted benzaldehyde is reacted with a substituted acetonitrile in a Knoevenagel-condensation reaction.

Description

Zimtsäurenitrile, Verfahren zu ihrer Herstellung und Verwendung Cinnamic acid nitriles, process for their preparation and use
Die benigne Prostatahyperplasie (BPH) ist mit Abstand die wichtigste urologische Erkrankung von Männern.Benign prostatic hyperplasia (BPH) is by far the most important urological disease in men.
Histologisch handelt es sich bei der BPH um ein benignes Wachstum von epithelialen und stromalen Anteilen der Prostata, bei dem es durch Vergrößerung des Organs zu Harnabflußstörungen kommt, die mit Symptomen wie Pollakisurie und Nykturie sowie unvollständiger und verzögerter Blasenentleerung einhergehen. In fortgeschrittenem Stadium kann es infolge der Rückstauung von Harn zu Niereninsuffizienz und Urämie kommen. Bedingt durch Sekretstauungen und Harnverhaltungen entwickeln sich außerdem häufig eine abakterielle Prostatitis, Kongestionen und rezidivierende Harnwegsinfektionen, die neben den obstruktiven Beschwerden für irritative Miktionsstörungen verantwortlich sind. Zunehmend wird deshalb auch von einem BPH-Symdrom oder "Lower Urinary Tract Symptoms (LUTS)" gesprochen. Ätiologie und Pathogenese der BPH sind noch weitgehend ungeklärt.Histologically, BPH is a benign growth of epithelial and stromal parts of the prostate, with enlargement of the organ leading to urinary drainage disorders that are accompanied by symptoms such as pollakiuria and nocturia, as well as incomplete and delayed bladder emptying. In advanced stages, urinary congestion can lead to renal failure and uremia. Due to secretion congestion and urinary retention, abacterial prostatitis, congestion and recurrent urinary tract infections often develop, which in addition to the obstructive symptoms are responsible for irritative micturition disorders. This is why there is increasing talk of a BPH symptom or "Lower Urinary Tract Symptoms (LUTS)". The aetiology and pathogenesis of BPH are still largely unknown.
Das Spektrum der Behandlungsmöglichkeiten der BPH reicht von einer abwartenden Haltung ("watchful waiting") bis zur offenen Prostataadenomektomie. Zunehmende Bedeutung gewinnen jedoch medikamentöse Therapieansätze. Neben Phytopharmaka, die häufig bei leichteren Krankheitsstadien eingesetzt werden, werden vor allem Alpha Antagonisten und 5-Alpha-Reduktasehemmer als Arzneimittel zur Behandlung der BPH verwendet (vgl. Eckert, R.E., et al., Zelluläre Grundlage der dynamischen, infravesikalen Obstruktion im Rahmen einer benignen Prostatahyperplasie; Rolle von Alpharezeptorenblockem und zyklischen Nukleotiden, Akt. Urol. 29, 252-260 (1998); Weckermann, D. und Wawroschek, F. Medikamentöse Therapie des benignen Prostatahyperplasie-Syndroms. Münch. med. Wschr. 141 , 54-58 (1999); Weckermann, D. und Wawroschek, F. Interventionelle Therapie der benignen Prostatahyperplasie. Münch. med. Wschr., 141 , 59-63 (1999).The spectrum of treatment options for BPH ranges from a watchful waiting to open prostate adenomectomy. However, drug therapy approaches are becoming increasingly important. In addition to phytopharmaceuticals, which are often used in milder stages of the disease, alpha antagonists and 5-alpha reductase inhibitors are primarily used as medicines for the treatment of BPH (see Eckert, RE, et al., Cellular basis of dynamic, infravesical obstruction in the context of a benign prostatic hyperplasia; role of alpha-receptor blockers and cyclic nucleotides, Akt. Urol. 29, 252-260 (1998); Weckermann, D. and Wawroschek, F. Medicinal therapy of benign prostatic hyperplasia syndrome. Münch. med. Wschr. 141, 54- 58 (1999); Weckermann, D. and Wawroschek, F. Interventional therapy for benign prostatic hyperplasia, Münch. Med. Wschr., 141, 59-63 (1999).
Der Einsatz von AlpharAntagonisten beruht auf der Ansicht, daß die Obstruktion der ableitenden Harnwege bei der BPH neben einer statischen Komponente, die durch die vergrößerte Prostata hervorgerufen wird, zusätzlich auf einer dynamischen Komponente beruht, die durch einem erhöhten Tonus der glatten prostatischen Muskulatur hervorgerufen wird. In einer Vielzahl von Studien wurde gezeigt, daß die dynamische Obstruktion bei der BPH offenbar durch eine gesteigerte Freisetzung von Noradrenalin aus sympathischen Neuronen vermittelt wird. Es ist heute generell akzeptiert, daß in der Prostata vorwiegend Alpha1A-Adrenozeptoren exprimiert werden. Die klinische Effektivität der Blockade von Alpha-Adrenozeptoren bei der Behandlung der BPH wurde ursprünglich durch Verwendung des unspezifischen Antagonisten Phenoxybenzamin demonstriert. Wegen der erheblichen kardiovaskulären Nebenwirkungen wurden in den vergangenen Jahren eine Reihe von sogenannten uroselektiven AlphaιA-Blockern entwickelt, die wesentlich besser verträglich sind. Die Ergebnisse von klinischen Studien zeigen, daß diese Substanzen im Vergleich zu Placebo eine statistisch und klinisch signifikante Verbesserung der Symptome und des maximalen Harnflusses bewirken. Vorteil der Alphablocker ist ihr rascher Wirkungseintritt, die Wachstumpotenz der Prostata bleibt durch diese Substanzen aber unbeeinflußt (vgl. Heimbach, D. und Müller, S.C. Die Behandlung der BPH mit α Adrenozeptorantagonisten. Urologe [A] 36, 18-34 (1997); Eckert et al., loc. cit.; Weckermann und Wawroschek, loc. cit.).The use of Alphar antagonists is based on the view that the obstruction of the urinary tract in BPH is based not only on a static component, which is caused by the enlarged prostate, but also on a dynamic component, which is caused by an increased tone of the smooth prostatic muscles. A large number of studies have shown that dynamic obstruction in BPH is apparently mediated by an increased release of norepinephrine from sympathetic neurons. It is generally accepted today that in the Prostate predominantly alpha 1A adrenoceptors are expressed. The clinical effectiveness of blocking alpha-adrenoceptors in the treatment of BPH was originally demonstrated using the non-specific antagonist phenoxybenzamine. Because of the significant cardiovascular side effects, a number of so-called uroselective alpha- A blockers have been developed in recent years, which are much better tolerated. The results of clinical studies show that these substances have a statistically and clinically significant improvement in symptoms and maximum urine flow compared to placebo. The advantage of alpha blockers is their rapid onset of action, but the growth potential of the prostate remains unaffected by these substances (cf. Heimbach, D. and Müller, SC. Treatment of BPH with α adrenoceptor antagonists. Urologe [A] 36, 18-34 (1997); Eckert et al., Loc. Cit .; Weckermann and Wawroschek, loc. Cit.).
Grundsätzlich entwickelt sich eine BPH nur in Anwesenheit biologisch aktiver männlicher Geschlechtshormone. Bei Männern, die sich vor dem 40. Lebensjahr einer Kastration unterziehen mußten oder bei denen aufgrund einer Unterfunktion der Hypophyse keine bzw. nur eine unzureichende Androgenbildung in den Gonaden erfolgt, wird praktisch nie eine BPH beobachtet. Ebenso unterbleibt bei einem angeborenen Defekt oder einem Fehlen von Androgenrezeptoren (z.B. testikuläre Feminisierung) die normale Entwicklung der Prostata und die Ausbildung hyperplastischer Veränderungen mit zunehmendem Alter. Das biologisch wichtigste Androgen in der Prostata ist Dihydrotestosteron (DHT), das lokal unter dem Einfluß der 5-Alpha-Reduktase aus Testosteron gebildet wird. Da DHT vor allem die epithelialen Anteile der BPH stimuliert, gelingt es durch Hemmung der 5-Alpha- Reduktase einen Wachstumsstillstand bzw. eine Atrophie des Drüsenanteils zu erreichen, die stromale Komponente der BPH wird aber praktisch nicht beeinflußt. Zum Beispiel kommt es bei der Einnahme des 5-Alpha-Reduktasehemmers Finasterid zu einer Reduzierung der prostatischen DHT-Konzentration von bis zu 85 %, die Verringerung der Prostatagröße beträgt durchschnittlich aber nur ca. 20 % und benötigt außerdem einen Zeitraum von bis zu 12 Monaten. Klinisch relevant ist dieser Effekt im wesentlichen nur dann, wenn das Prostatavolumen zu Beginn der Therapie bei über 40 ml liegt (vgl. Geller, J. Pathogenesis and medical treatment of benign prostatic hyperplasia. Prostate (Suppl.) 2, 95-104 (1989); Weckermann und Wawroschek, loc. cit.).Basically, BPH only develops in the presence of biologically active male sex hormones. BPH is practically never observed in men who had to undergo castration before the age of 40 or who have no or only insufficient androgen formation in the gonads due to an underactive pituitary gland. Likewise, in the event of a congenital defect or a lack of androgen receptors (e.g. testicular feminization), the normal development of the prostate and the formation of hyperplastic changes with increasing age do not occur. The most biologically important androgen in the prostate is dihydrotestosterone (DHT), which is produced locally under the influence of 5-alpha reductase from testosterone. Since DHT mainly stimulates the epithelial parts of BPH, inhibition of 5-alpha reductase leads to growth arrest or atrophy of the glandular part, but the stromal component of BPH is practically unaffected. For example, taking the 5-alpha reductase inhibitor finasteride results in a reduction in the prostatic DHT concentration of up to 85%, but the reduction in prostate size is only about 20% on average and also requires a period of up to 12 months , This effect is only clinically relevant if the prostate volume at the beginning of therapy is over 40 ml (see Geller, J. Pathogenesis and medical treatment of benign prostatic hyperplasia. Prostate (Suppl.) 2, 95-104 (1989 ); Weckermann and Wawroschek, loc.cit.).
Obwohl Androgenen eine zentrale Rolle sowohl bei der normalen Entwicklung und Funktion der Prostata als auch bei der Ausbildung einer BPH zukommt, sind männliche Geschlechtshormone für sich alleine nicht ausreichend, um ein Wachstum von Prostatazellen auszulösen. Zahlreiche experimentelle Untersuchungen lassen vermuten, daß die wachstumsstimulierenden Effekte von Androgenen in vivo über die lokale Synthese von Wachstumsfaktoren vermittelt werden, und daß Störungen der para- und autokrinen Mechanismen der Wachstumskontrolle innerhalb der epithelialen und stromalen Anteile der Prostata maßgeblich an der Entstehung der BPH beteiligt sind. Tatsächlich wurden in der Prostata eine Vielzahl von Wachstumsfaktoren (z.B. Epidermal Growth Factor [EGFJ, Transforming Growth Factor-α, [TGF-α], TGF-ß, basic Fibroblast Growth Factor (bFGF), Keratinocyte Growth Factor [KGF], Nerve Growth Factor [NGF], Insulin-like Growth Factor I [IGF-1] etc.) und deren Rezeptoren nachgewiesen. Da die BPH sehr häufig mit entzündlichen Reaktionen einhergeht, denen eine wesentliche Rolle bei der Pathogenese beigemessen wird, kommt Platelet-derived Growth Factor (PDGF), der z.B. von Fibroblasten, Thrombozyten und Leukozyten freigesetzt wird, vermutlich eine besondere Bedeutung für die Proliferation von Prostatazellen zu (vgl. Vlahos, C.J., et al., Platelet- derived growth factor induces proliferation of hyperplastic human prostatic stromal cells. J. Cell. Biochem. 52, 404-413 (1993); Desgrandchamps, F. und Teillac, P. The role of growth factors in the pathogenesis of benign prostatic hyperplasia. Biomed. Pharmacother. 48 (Suppl. 1), 19s-23s (1994); Sciarra, F., et al., Regional distribution of epidermal growth factor, testosterone and dihydrotestosterone in benign prostatic hyperplasia tissue. Urol. Res. 23, 387-390 (1995); Claesson-Welsh, L. Mechanism of action of platelet-derived growth factor. Int. J. Biochem. Cell Biol. 28, 373-385 (1996); Culig, Z., et al., Regulation of prostatic growth and function by peptide growth factors. Prostate 28, 392-405 (1996); Peehl, D.M. Celluiar biology of prostatic growth factors. Prostate (Suppl.) 6, 74-78 (1996); Peehl, D.M. und Seilers, R.G. Basic FGF, EGF, and PDGF modify TGFß-induction of smooth muscle cell phenotype in human prostatic stromal cells. Prostate 35, 125-134 (1998).Although androgens play a central role in the normal development and function of the prostate as well as in the formation of BPH, male sex hormones alone are not sufficient to trigger the growth of prostate cells. Numerous experimental investigations suggest that the growth-stimulating effects of androgens are mediated in vivo via the local synthesis of growth factors, and that disorders of the para- and autocrine mechanisms of growth control within the epithelial and stromal parts of the prostate are significantly involved in the development of BPH , In fact, a large number of growth factors (e.g. epidermal growth factor [EGFJ, transforming growth factor-α, [TGF-α], TGF-ß, basic fibroblast growth factor (bFGF), keratinocyte growth factor [KGF], nerve growth Factor [NGF], Insulin-like Growth Factor I [IGF-1] etc.) and their receptors. Since BPH is very often associated with inflammatory reactions, which are considered to play an important role in pathogenesis, platelet-derived growth factor (PDGF), which is released by fibroblasts, thrombocytes and leukocytes, is probably of particular importance for the proliferation of prostate cells to (see Vlahos, CJ, et al., Platelet-derived growth factor induces proliferation of hyperplastic human prostatic stromal cells. J. Cell. Biochem. 52, 404-413 (1993); Desgrandchamps, F. and Teillac, P. The role of growth factors in the pathogenesis of benign prostatic hyperplasia. Biomed. Pharmacother. 48 (Suppl. 1), 19s-23s (1994); Sciarra, F., et al., Regional distribution of epidermal growth factor, testosterone and dihydrotestosterone in benign prostatic hyperplasia tissue. Urol. Res. 23, 387-390 (1995); Claesson-Welsh, L. Mechanism of action of platelet-derived growth factor. Int. J. Biochem. Cell Biol. 28, 373-385 ( 1996); Culig, Z., et al., Regulation of pro static growth and function by peptide growth factors. Prostate 28: 392-405 (1996); Peehl, DM Celluiar biology of prostatic growth factors. Prostate (Suppl.) 6: 74-78 (1996); Peehl, DM and Seilers, RG Basic FGF, EGF, and PDGF modify TGFß-induction of smooth muscle cell phenotype in human prostatic stromal cells. Prostate 35, 125-134 (1998).
Wachstumsfaktoren vermitteln ihre biologische Wirkung durch Bindung an spezifische Rezeptoren an der Zelloberfläche, die über intrinsische Tyrosinkinase-Aktivität verfügen. Nach Bindung des Liganden kommt es zur Phosphorylierung von Tyrosinresten an der intrazellulären Rezeptordomäne und anderer Zielproteine, die anschließend eine Kaskade von intrazellulären Reaktionen auslöst. Dazu gehören die Stimulation der Protein- und DNA-Synthese sowie die Aktivierung der Zellproliferation. Inhibitoren von Rezeptor- Tyrosinkinase werden deshalb als vielversprechende Substanzen zur Entwicklung neuer Medikamente zur Behandlung von Erkrankungen angesehen, die mit einer erhöhten Zellproliferation einhergehen (z.B. Krebs, Atherosklerose, Psoriasis) (vgl. Levitzki, A. und Gazit, A. Tyrosine kinase Inhibition: An approach to drug development. Science 267, 1782- 1788 (1995); Traxler, P. und Lydon, N. Recent advances in protein tyrosine kinase inhibitors. Drugs Fut. 20, 1261-1274 (1995)). Wenig Aufmerksamkeit wurde diesem Wirkprinzip bisher aber bei der Therapie der BPH entgegengebracht.Growth factors mediate their biological effect by binding to specific receptors on the cell surface that have intrinsic tyrosine kinase activity. After binding of the ligand, tyrosine residues on the intracellular receptor domain and other target proteins are phosphorylated, which subsequently triggers a cascade of intracellular reactions. This includes stimulating protein and DNA synthesis and activating cell proliferation. Inhibitors of receptor tyrosine kinase are therefore seen as promising substances for the development of new drugs for the treatment of diseases that are associated with increased cell proliferation (e.g. cancer, atherosclerosis, psoriasis) (see Levitzki, A. and Gazit, A. Tyrosine kinase inhibition: An approach to drug development. Science 267, 1782-1788 (1995); Traxler, P. and Lydon, N. Recent advances in protein tyrosine kinase inhibitors. Drugs Fut. 20, 1261-1274 (1995)). So far, little attention has been paid to this principle of action in the treatment of BPH.
Der Erfindung liegt deshalb die Aufgabe zugrunde, Substanzen zur Verfügung zu stellen, die durch Hemmung von AlpharAdrenozeptoren (αi-antagonistische Wirkung) und durch Unterdrückung derWachstumsfaktor-vermittelten Zellproliferation (proliferations-hemmende Wirkung) von Epithel- und Stromazellen sowohl die dynamische als auch die statische Komponente der BPH positiv beeinflussen und damit eine umfassende Behandlung des BPH-Syndroms erlauben. Aufgabe der Erfindung ist ferner die Bereitstellung von Arzneimitteln zur Prophylaxe und Behandlung der BPH und von Verfahren zu deren Herstellung und Verwendung.The invention is therefore based on the object of providing substances which, by inhibiting alphar adrenoceptors (αi-antagonistic action) and by suppressing the growth factor-mediated cell proliferation (proliferation-inhibiting action) of epithelial and stromal cells, both the dynamic and the static Affect components of BPH positively and thus allow comprehensive treatment of BPH syndrome. The object of the invention is also the provision of medicaments for the prophylaxis and treatment of BPH and of processes for their production and use.
Diese Aufgabe wird erfindungsgemäß gelöst durch substituierte Zimtsäurenitrile der allgemeinen Formel I,
Figure imgf000006_0001
According to the invention, this object is achieved by substituted cinnamic acid nitriles of the general formula I,
Figure imgf000006_0001
worin die Substituenten folgende Bedeutungen besitzen:where the substituents have the following meanings:
X = CN, COOR4, CONR5R6, COAryl, COAlkyl, Aryl, Heteroaryl,X = CN, COOR 4 , CONR 5 R 6 , COAryl, COalkyl, aryl, heteroaryl,
R1 = H, Alkyl, Arylalkyl, [4-(2-Alkoxyphenyl)-1-piperazinyl]alkyl,R 1 = H, alkyl, arylalkyl, [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl,
R2 und R3 = unabhängig voneinander H, Hydroxy, Alkoxy, Benzyl,R 2 and R 3 = independently of one another H, hydroxy, alkoxy, benzyl,
R = H, Alkyl, [4-(2-Alkoxyphenyl)-1-piperazinyl]alkyl,R = H, alkyl, [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl,
R5 = H, Alkyl, Alkoxyalkyl, Alkoxy-oxoalkyl, Dialkylaminoalkyl, Amino-oxoalkyl, Aryl,R 5 = H, alkyl, alkoxyalkyl, alkoxy-oxoalkyl, dialkylaminoalkyl, amino-oxoalkyl, aryl,
Arylalkyl, [4-(2-Alkoxyphenyl)-1-piperazinyl]alkyl, Morpholinoalkyl, Furanylalkyl,Arylalkyl, [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl, morpholinoalkyl, furanylalkyl,
Thienylalkyl, R6 = H, Alkyl oder NR5R6 gemeinsam = cyclisches Amin wie Pyrrolidin, Morpholin, Piperidin, subst. Piperazin, wobei mindestens einer der Reste R1 , R4 oder R5 ein [4-(2-Alkoxyphenyl)-1- piperazinyl]alkyl-Rest ist.Thienylalkyl, R 6 = H, alkyl or NR 5 R 6 together = cyclic amine such as pyrrolidine, morpholine, piperidine, subst. Piperazine, where at least one of the radicals R 1 , R 4 or R 5 is a [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl radical.
Bei den vorstehenden Definitionen der allgemeinen Formel I ist unter "Alkyl" in allen Fällen ein geradkettiger oder verzweigter Alkylrest mit 1 - 8 C-Atomen, vorzugsweise mit 1 - 4 C- Atomen, und unter "Alkoxy" eine Alkoxygruppe mit 1 - 4 C-Atomen, vorzugsweise mit einem oder zwei C-Atomen, zu verstehen. Unter „Aryl" sind auch ggf. substituierte Arylreste zu verstehen, beispielsweise Halogenphenyl, Alkylphenyl, Alkoxyphenyl,In the above definitions of the general formula I, "alkyl" in all cases is a straight-chain or branched alkyl radical having 1 to 8 carbon atoms, preferably having 1 to 4 carbon atoms, and "alkoxy" is an alkoxy group having 1 to 4 carbon atoms Atoms, preferably with one or two carbon atoms. “Aryl” is also to be understood as meaning any substituted aryl radicals, for example halophenyl, alkylphenyl, alkoxyphenyl,
Trifluormethylphenyl. Dies gilt sinngemäß auch für „COAryl", „Arylalkyl", „Heteroaryl" (= Arylrest, worin mindestens ein Ring-C-Atom durch ein Heteroatom, z. B. Stickstoff im Falle des Pyridins, ersetzt ist), etc.Trifluoromethylphenyl. This also applies analogously to "COAryl", "Arylalkyl", "Heteroaryl" (= aryl radical, in which at least one ring carbon atom is replaced by a heteroatom, for example nitrogen in the case of pyridine), etc.
Da die erfindungsgemäßen Verbindungen durch Reste oder Gruppen substituiert sind, die basische Stickstoffatome enthalten, beispielsweise cyclische Amingruppen wie Pyrrolidin, Morpholin, Piperidin, Piperazin, können diese Verbindungen entweder in Form freier Basen oder aber in Form von Additionssalzen mit pharmakologisch verträglichen Säuren vorliegen, beispielsweise in Form ihrer Hydrochloride oder Trifluoroacetate.Since the compounds according to the invention are substituted by radicals or groups which contain basic nitrogen atoms, for example cyclic amine groups such as pyrrolidine, morpholine, piperidine, piperazine, these compounds can be present either in the form of free bases or in the form of addition salts with pharmacologically acceptable acids, for example in Form of their hydrochloride or trifluoroacetate.
Bevorzugte Zimtsäurenitril-Derivate sind diejenigen Verbindungen der allgemeinen Formen I, bei denen der mindestens eine [4-(2-Alkoxyphenyl)-1-piperazinyl]alkyl-Rest ein [4-(2- Methoxyphenyl)-1-piperazinyl]propyl-Rest ist.Preferred cinnamic acid nitrile derivatives are those compounds of general forms I in which the at least one [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl radical is a [4- (2-methoxyphenyl) -1-piperazinyl] propyl radical is.
Die erfindungsgemäßen Verbindungen sind pharmakologisch wirksam; ihre pharmakologische Wirkung ist gekennzeichnet durch das gleichzeitige Vorhandensein einer αTAdrenozeptoren-antagonistischen Wirkungskomponente und einer proliferationshemmenden Wirkungskomponente. Gegenstand der Erfindung sind deshalb auch Arzneimittel, die als Wirkstoff, neben üblichen Hilfs- und Zusatzstoffen, eine für die Prophylaxe oder Behandlung der BPH wirksame Menge an einer oder mehreren der erfindungsgemäßen Verbindungen der allgemeinen Formen I enthalten.The compounds according to the invention are pharmacologically active; Their pharmacological effect is characterized by the simultaneous presence of an α T adrenoceptor-antagonistic active component and a proliferation-inhibiting active component. The invention therefore also relates to medicaments which, as an active ingredient, contain, in addition to customary auxiliaries and additives, an amount of one or more of the compounds of general forms I according to the invention which is effective for the prophylaxis or treatment of BPH.
Als pharmakologisch inerte Hilfsstoffe können diese Arzneimittel zum Beispiel Wasser, pflanzliche Öle, Polyethylenglykole, Glycerinester, Gelatine, Kohlenhydrate wie Lactose oder Stärke, Magnesiumstearat, Talk, Vaseline®, Konservierungsmittel, Netzmittel, Emulgatoren, physiologisch unbedenkliche Salze, Puffersubstanzen, Farbstoffe, Geschmacksstoffe und Aromastoffe enthalten. Die Wahl der Hilfsstoffe hängt von der gewünschten Applikationsform wie z. B. Tabletten, Dragees, Säfte, Ampullen, Suppositorien, Salben oder Sprays ab. Die erfindungsgemäßen Verbindungen können auch im Gemisch mit anderen bekannten Wirkstoffen verabreicht werden.As pharmacologically inert adjuvants, these medicaments can include, for example, water, vegetable oils, polyethylene glycols, glycerol esters, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, Vaseline®, preservatives, wetting agents, emulsifiers, physiologically acceptable salts, buffer substances, colorants, flavorings and flavorings contain. The choice of excipients depends on the desired application form such as. B. tablets, dragees, juices, ampoules, suppositories, ointments or sprays. The compounds according to the invention can also be administered in a mixture with other known active compounds.
Die Verbindung (E)-1 -[2-Cyano-3-(3,4-dihydroxyphenyl)-1 -oxo-2-propenyl]-4-phenyl- piperazin ist bekannt aus A. Gazit et al., J. Med. Chem. 34, S. 1896-1907 (1991) (dort Verbindung Nr. 68). Diese bekannte Verbindung unterscheidet sich von den erfindungsgemäßen Verbindungen der allgemeinen Formel I dadurch, daß sie zwar einen [4-Phenyl-1-piperazinyl]-Rest enthält, nicht aber mindestens einen [4-(2-Alkoxyphenyl)-1 - piperazinyl]alkyl-Rest. Für die bekannte Verbindung wird zwar eine proliferations- hemmende Wirkung beschrieben, es ist jedoch nicht bekannt, ob diese Verbindung auch eine αi-antagonistische Wirkung besitzt.The compound (E) -1 - [2-cyano-3- (3,4-dihydroxyphenyl) -1-oxo-2-propenyl] -4-phenylpiperazine is known from A. Gazit et al., J. Med Chem. 34, pp. 1896-1907 (1991) (there compound no. 68). This known compound differs from the compounds of the general formula I according to the invention in that it contains a [4-phenyl-1-piperazinyl] radical, but not at least one [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl -Rest. Although a proliferation-inhibiting effect is described for the known compound, it is not known whether this compound also has an αi-antagonistic effect.
Gegenstand der Erfindung sind schließlich auch Verfahren zur Herstellung der Verbindungen der allgemeinen Formel I, bei denen ein Benzaldehyd der allgemeinen Formel II, worin R1, R2 und R3 die oben angegebenen Bedeutungen besitzen, mit einem substituierten Acetonitril der allgemeinen Formel III, worin X die oben angegebene Bedeutung besitzt, unter Bedingungen umgesetzt wird, die für Knoevenagel- Kondensationen, d.h. Aldolkondensationen, bekannt sind.The invention finally also relates to processes for the preparation of the compounds of the general formula I in which a benzaldehyde of the general formula II, in which R 1 , R 2 and R 3 have the meanings given above, with a substituted acetonitrile of the general formula III, in which X has the meaning given above, is implemented under conditions known for Knoevenagel condensations, ie aldol condensations.
Figure imgf000007_0001
Figure imgf000007_0001
( II ) ( III ) - H2O(II) (III) - H 2 O
Dabei kann die Knoevenagel-Kondensation mit oder ohne Katalysator durchgeführt werden. Als Katalysatoren dienen beispielsweise Piperidin, Ammoniumacetat, ß-Alanin oder Eisessig.The Knoevenagel condensation can be carried out with or without a catalyst. Piperidine, ammonium acetate, β-alanine or glacial acetic acid serve as catalysts.
Die Verbindungen der allgemeinen Formel II werden, soweit sie nicht bekannt und / oder kommerziell erhältlich sind, durch Alkylierung des entsprechend mit R1, R2 und R3 substituierten Benzaldehyds, worin R1 = H ist und R2 und R3 die oben angegebenen Bedeutungen haben, mit einem Halogenid R1Hal oder einem Sulfonsäureester R1OSO2Alkyl bzw. R1OSO2Aryl hergestellt.Insofar as they are not known and / or commercially available, the compounds of the general formula II are obtained by alkylation with R 1 , R 2 and R 3 Substituted benzaldehyde, in which R 1 = H and R 2 and R 3 have the meanings given above, with a halide R 1 Hal or a sulfonic ester R 1 OSO 2 alkyl or R 1 OSO 2 aryl.
Diejenigen Verbindungen der allgemeinen Formel II, worinThose compounds of the general formula II in which
R1 = [4-(2-Alkoxyphenyl)-1-piperazinyl]alkylR 1 = [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl
R2 und R3 = unabhängig voneinander H, Hydroxy, Alkoxy mit 1 - 4, vorzugsweise 1 - 2 C-R 2 and R 3 = independently of one another H, hydroxy, alkoxy with 1 - 4, preferably 1 - 2 C-
Atomen, Benzyl, bedeuten, sind neue reaktive Zwischenprodukte zur Herstellung der Verbindungen der allgemeinen Formel I, worin R1, R2 und R3 die entsprechenden Bedeutungen besitzen. Diese Zwischenprodukte sind ebenfalls Gegenstand der Erfindung.Atoms, benzyl, are new reactive intermediates for the preparation of the compounds of general formula I, in which R 1 , R 2 and R 3 have the corresponding meanings. These intermediates are also the subject of the invention.
Verbindungen der allgemeinen Formel III mit X = CONR5R6 werden durch Umsetzung eines Esters der allgemeinen Formel III mit X = COOR4, worin R4 Methyl oder Ethyl bedeutet, mit dem entsprechenden Amin HNR5R6 hergestellt. Ester der allgemeinen Formel III mit X = COOR4 mit R4 = [4-(2-Alkoxyphenyl)-1-piperazinyl]alkyl erhält man durch Umesterung des entsprechenden Methylesters mit dem Alkohol R4OH.Compounds of the general formula III with X = CONR 5 R 6 are prepared by reacting an ester of the general formula III with X = COOR 4 , in which R 4 is methyl or ethyl, with the corresponding amine HNR 5 R 6 . Esters of the general formula III with X = COOR 4 with R 4 = [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl are obtained by transesterification of the corresponding methyl ester with the alcohol R 4 OH.
Die Erfindung wird nachfolgend anhand von Beispielen und pharmakologischen Untersuchungen näher erläutert.The invention is explained in more detail below with the aid of examples and pharmacological studies.
Die im nachfolgenden Text verwendeten Abkürzungen bedeuten: TBME = tert.-Butyl- Methylether und PE = Petrolether.The abbreviations used in the following text mean: TBME = tert-butyl methyl ether and PE = petroleum ether.
I. Beispiele 1 bis 72 für Endprodukte der allgemeinen Formel II. Examples 1 to 72 for end products of the general formula I
Zur Herstellung der in den nachfolgenden Beispielen 1 bis 72 näher beschriebenen Verbindungen wurden folgende Verfahren benutzt:The following processes were used to prepare the compounds described in more detail in Examples 1 to 72 below:
Verfahren A: Der entsprechend mit R1 bis R3 substituierte Benzaldehyd der allgemeinen Formel II, gegebenenfalls aus seinem Hydrochlorid freigesetzt, 0.9 bis 2.0 Äquivalente des entsprechend mit X substituierten Acetonitrils der allgemeinen Formel III und ggf. eine katalytische Menge Piperidin werden in Ethanol, Methanol, 2-Propanol oder TBME 2 h bis 3 d bei 50 °C bis Siedetemperatur gerührt. Die Aufarbeitung erfolgt durch Abfiltrieren des Produkts, durch Eindampfen oder durch Verteilung zwischen Ethylacetat oder Chloroform und Wasser. Gereinigt wird durch Chromatographie, Umkristallisation und/oder Salzbildung nach Verfahren C.Process A: The benzaldehyde of the general formula II correspondingly substituted with R 1 to R 3 , if appropriate released from its hydrochloride, 0.9 to 2.0 equivalents of the acetonitrile of the general formula III substituted with X and optionally a catalytic amount of piperidine are dissolved in ethanol, methanol , 2-propanol or TBME 2 h to 3 d at 50 ° C to boiling temperature. The product is worked up by filtering off the product, by evaporation or by partitioning between ethyl acetate or chloroform and water. Purification is carried out by chromatography, recrystallization and / or salt formation using method C.
Verfahren B: Der entsprechend mit R1 bis R3 substituierte Benzaldehyd der allgemeinen Formel II, gegebenenfalls aus seinem Hydrochlorid freigesetzt, 0.9 bis 1.1 Äquivalente des entsprechend mit X substituierten Acetonitrils der allgemeinen Formel III und gegebenenfalls eine katalytische Menge Piperidin und/oder Eisessig werden in Toluol 2 bis 24 h am Wasserabscheider unter Rückfluß gerührt. Die Aufarbeitung erfolgt durch Eindampfen der Reaktionslösung. Gereinigt wird durch Chromatographie, Umkristallisation und/oder Salzbildung nach Verfahren C.Process B: The benzaldehyde of the general formula II substituted with R 1 to R 3 , if appropriate released from its hydrochloride, 0.9 to 1.1 equivalents of the Acetonitrile of the general formula III corresponding to X and optionally a catalytic amount of piperidine and / or glacial acetic acid are stirred in toluene for 2 to 24 hours on a water separator under reflux. The mixture is worked up by evaporating the reaction solution. Purification is carried out by chromatography, recrystallization and / or salt formation using method C.
Verfahren C: Die nach dem Verfahren A oder B hergestellte Base wird in Ethanol, 2- Propanol oder Aceton gelöst und mit 1 bis 3 Äquivalenten einer Lösung von Chlorwasserstoff in 2-Propanol oder wässriger Salzsäure versetzt. Man saugt das Hydrochlorid ab oder engt ein und reinigt durch Umkristallisation.Process C: The base prepared by process A or B is dissolved in ethanol, 2-propanol or acetone and 1 to 3 equivalents of a solution of hydrogen chloride in 2-propanol or aqueous hydrochloric acid are added. The hydrochloride is suctioned off or concentrated and purified by recrystallization.
Verfahren D: Der entsprechend mit R1 bis R3 substituierte tert.-Butylester der allgemeinen Formel I (X = COOC(CH3)3) wird in Trifluoressigsäure einige Stunden bei Raumtemperatur gerührt. Das Reaktionsgemisch wird einrotiert und der Rückstand umkristallisiert. Process D: The tert-butyl ester of the general formula I (X = COOC (CH 3 ) 3 ) correspondingly substituted by R 1 to R 3 is stirred in trifluoroacetic acid at room temperature for a few hours. The reaction mixture is spun in and the residue is recrystallized.
Figure imgf000010_0001
Figure imgf000010_0001
* *
Figure imgf000011_0001
Figure imgf000012_0001
Figure imgf000011_0001
Figure imgf000012_0001
to to
Figure imgf000013_0001
Figure imgf000013_0001
II II
Figure imgf000014_0001
Figure imgf000014_0001
M M
Figure imgf000015_0001
Figure imgf000015_0001
li li
Figure imgf000016_0001
Figure imgf000016_0001
m
Figure imgf000017_0001
m
Figure imgf000017_0001
/S / S
Figure imgf000018_0001
Figure imgf000018_0001
II. Beispiele 73 bis 83 für Vorprodukte der allgemeinen Formel IIII. Examples 73 to 83 for precursors of the general formula II
Zur Herstellung der in den nachfolgenden Beispielen 73 bis 83 näher beschriebenen Verbindungen wurde folgendes Verfahren benutzt:The following procedure was used to prepare the compounds described in more detail in Examples 73 to 83 below:
Verfahren E: Zu 1.0 bis 1.2 Äquivalenten des entsprechend substituierten 1-(3- Chlorpropyl)-4-(2-alkoxyphenyl)piperazin Dihydrochlorids in Ethanol oder 1-Propanol werden 2.0 bis 2.4 Äquivalente Kaliumhydroxid gegeben. Nach Zugabe von 0.1 bis 0.4 Äquivalenten Kaliumiodid, 1.0 bis 1.2 Äquivalenten Kaliumcarbonat und 0.9 bis 1.0 Äquivalenten des entsprechend mit R2 bis R3 substituierten Benzaldehyds (R = H) der allgemeinen Formel II wird 2 bis 24 h bei Siedetemperatur unter Rückfluß gerührt. Das anorganische Material wird abfiltriert und das Filtrat einrotiert. Die Aufarbeitung erfolgt durch Extraktion mit Ethylacetat oder TBME und erneuter Filtration und/oder durch Verteilung zwischen Ethylacetat oder TBME und Wasser. Gereinigt wird durch Umkristallisation oder Salzbildung nach Verfahren C.Method E: 2.0 to 2.4 equivalents of potassium hydroxide are added to 1.0 to 1.2 equivalents of the appropriately substituted 1- (3-chloropropyl) -4- (2-alkoxyphenyl) piperazine dihydrochloride in ethanol or 1-propanol. After adding 0.1 to 0.4 equivalents of potassium iodide, 1.0 to 1.2 equivalents of potassium carbonate and 0.9 to 1.0 equivalents of the benzaldehyde (R = H) of the general formula II substituted by R 2 to R 3 , the mixture is stirred under reflux at the boiling temperature for 2 to 24 h. The inorganic material is filtered off and the filtrate is spun in. Working up is carried out by extraction with ethyl acetate or TBME and renewed filtration and / or by partitioning between ethyl acetate or TBME and water. It is cleaned by recrystallization or salt formation according to method C.
Verfahren F: 1.0 Äqivalent Isovanillin, 1.5 Äquivalente 1-Brom-2-chlorethan bzw. 1 ,4- Dibrombutan und 1.5 Äquivalente Kaliumcarbonat werden in 2-Butanon 4 bis 20 h bei Siedetemperatur gerührt. Das anorganische Material wird abfiltriert und das Filtrat einrotiert. Die Reinigung erfolgt durch Chromatographie und/oder Umkristallisation. 1.0 Äquivalente des so hergestellten 3-(Haiogenalkoxy)-4-methoxybenzaldehyds, 1.0 Äquivalent 1-(2-Methoxyphenyl)piperazin, 1.5 bis 2.0 Äquivalente Kaliumhydrogen-carbonat und 0.1 Äquivalent Kaliumiodid werden in Dimethylformamid 9 bis 20 h bei 70 bis 80 °C gerührt Das anorganische Material wird abfiltriert und das Filtrat einrotiert Die Aufarbeitung erfolgt durch Extraktion mit Ethylacetat und erneuter Filtration oder durch Verteilung zwischen Ethylacetat und Wasser Gereinigt wird durch Chromatographie und Salzbildung nach Verfahren CProcess F: 1.0 equivalent of isovanillin, 1.5 equivalents of 1-bromo-2-chloroethane or 1, 4-dibromobutane and 1.5 equivalents of potassium carbonate are stirred in 2-butanone at boiling temperature for 4 to 20 h. The inorganic material is filtered off and the filtrate is spun in. The purification is carried out by chromatography and / or recrystallization. 1.0 equivalents of the 3- (Haiogenalkoxy) -4-methoxybenzaldehyde thus produced, 1.0 equivalent of 1- (2-methoxyphenyl) piperazine, 1.5 to 2.0 equivalents of potassium hydrogen carbonate and 0.1 equivalent of potassium iodide are in dimethylformamide at 70 to 80 ° C for 9 to 20 h The inorganic material is filtered off and the filtrate is spun in. Working up is carried out by extraction with ethyl acetate and renewed filtration or by partitioning between ethyl acetate and water. Purification is carried out by chromatography and salt formation according to process C.
Figure imgf000019_0001
Figure imgf000019_0001
I) III. Beispiele 84 bis 95 für Vorprodukte der allgemeinen Formel IIII) III. Examples 84 to 95 for precursors of the general formula III
Zur Herstellung der in den nachfolgenden Beispielen 84 bis 95 näher beschriebenen Verbindungen wurde folgendes Verfahren benutzt:The following procedure was used to prepare the compounds described in more detail in Examples 84 to 95 below:
Verfahren G: Das Amin HNR5R6, gegebenenfalls aus seinem Hydrochlorid freigesetzt, wird mit 1.0 bis 1.05 Äquivalenten Cyanessigsäuremethylester in Methanol, Ethanol oder ohne Lösungsmittel 15 min bis 28 h bei Raumtemperatur bis 100 °C umgesetzt. Nach dem Abkühlen wird gegebenenfalls einrotiert und durch Umkristallisation gereinigt.Process G: The amine HNR 5 R 6 , optionally released from its hydrochloride, is reacted with 1.0 to 1.05 equivalents of methyl cyanoacetate in methanol, ethanol or without solvent for 15 min to 28 h at room temperature to 100 ° C. After cooling, the mixture is optionally rotated in and purified by recrystallization.
Verfahren H: Der Alkohol HÖR4 wird mit 5 Äquivalenten Cyanessigsäuremethylester 7 h bei leichtem Unterdruck auf 80 °C erhitzt. Nach dem Abkühlen wird durch Säulenchromatographie und nachfolgende Umkristallisation gereinigt.Method H: The alcohol HÖR 4 is heated with 5 equivalents of methyl cyanoacetate to 80 ° C. for 7 hours at a slightly reduced pressure. After cooling, it is purified by column chromatography and subsequent recrystallization.
Figure imgf000020_0001
Figure imgf000020_0001
I* I *
Figure imgf000021_0001
Figure imgf000021_0001
Pharmakologische UntersuchungenPharmacological examinations
A Die pharmakolgische Prüfung der Substanzen auf Wechselwirkung mit Alpha^ Adrenozeptoren erfolgte mit einem Rezeptorbindungsassay unter Verwendung von Ratten-Gehimzellmembranen. Zur Praparation der Zellmembranen wurden mannliche Sprague-Dawley-Ratten (150-250 g) in CO2-Narkose euthanasiert und die Gehirne (ohne das Kleinhirn) entnommen Nach Entfernung von anhaftendem Blut und Hirnhauten wurden die Gehirne unmittelbar anschließend in einem 10fachen Volumen eiskaltem Homogenisierungspuffer (50 mM Tris-HCI, pH 7,4) aufgenommen und mit einem eisgekühlten Glashomogenisator homogenisiert. Das Zellhomogenat wurde für 10 min bei 50.000 g (4 βC) zentrifugiert und das Pellet in eiskaltem Homogenisierungspuffer resuspendiert. Nach erneuter Zentnfugation (10 min bei 4 °C und 50.000 g) wurden die Zellmembranen in einem 10fachen Volumen Bindungspuffer (50 mM Tris-HCI, 0,5 mM Na-EDTA, 0,01 % Ascorbinsaure, 10 μM Pargylin, pH 7,4) aufgenommen und in Portionen (1 ml) bei -80°C aufbewahrtA The pharmacological testing of the substances for interaction with alpha-adrenoceptors was carried out using a receptor binding assay using rat brain cell membranes. For the preparation of the cell membranes, male Sprague-Dawley rats (150-250 g) were euthanized in CO 2 anesthesia and the brains (without the cerebellum) were removed. After removal of adhering blood and cerebral membranes, the brains were immediately removed in a 10-fold volume of ice-cold homogenization buffer (50 mM Tris-HCl, pH 7.4) and homogenized with an ice-cooled glass homogenizer. The cell homogenate was centrifuged for 10 min at 50,000 g (4 β C) and the pellet resuspended in ice-cold homogenization buffer. After renewed centrifugation (10 min at 4 ° C. and 50,000 g), the cell membranes were in a 10-fold volume of binding buffer (50 mM Tris-HCl, 0.5 mM Na-EDTA, 0.01% ascorbic acid, 10 μM pargyline, pH 7, 4) and stored in portions (1 ml) at -80 ° C
Für den Rezeptorbindungsassay wurden die Substanzen unter Verwendung von DMSO in 150 μl Bindungspuffer gelost und zusammen mit 50 μl Gehirnzellmembranen (2,5 mg/ml Protein) und 50 μl 3H-Prazosιn (300 pM, spez. Aktivität 80 Ci/mmol) in Bindungspuffer für 45 min bei Raumtemperatur inkubiert. Die Bestimmung der unspezifischen Bindung erfolgte in Anwesenheit von 2 μM Phentolamin. Die Reaktionsmischung wurde anschließend durch Glasfaserfiiter (Typ GF/B), die über Nacht mit Polyethylenimine (0,2 % in Agua dest.) vorbehandelt worden waren, gefiltert. Nach viermaligem Waschen mit jeweils 3 ml eiskaltem Bindungspuffer wurden die Filter für 1 h bei 60 °C getrocknet. Die Bestimmung der gebundenen Radioaktivität erfolgte nach Überführung der Filter in 4 ml Szintillationsflussigkeit (Quickszint) in einem Beta-Counter. Die Hemmung der spezifischen Bindung von 3H-Prazosιn an Alpha1-Adenozeptoren wurde in Prozent einer parallel untersuchten Losungsmittelkontrolle kalkuliertFor the receptor binding assay, the substances were dissolved in 150 μl binding buffer using DMSO and together with 50 μl brain cell membranes (2.5 mg / ml protein) and 50 μl 3 H-Prazosιn (300 pM, specific activity 80 Ci / mmol) in Binding buffer incubated for 45 min at room temperature. The non-specific binding was determined in the presence of 2 μM phentolamine. The reaction mixture was then filtered through glass fiber filters (type GF / B) which had been pretreated overnight with polyethyleneimine (0.2% in distilled water). After washing four times with 3 ml of ice-cold binding buffer, the filters were dried at 60 ° C. for 1 h. The radioactivity bound was determined after transferring the filter to 4 ml of scintillation liquid (Quickszint) in a beta counter. The inhibition of the specific binding of 3 H-Prazosιn to Alpha 1 adenoceptors was calculated in percent of a solvent control examined in parallel
B Der Einfluß der Substanzen auf die Wachstumsfaktor-induzierte Zellproliferation wurde an NIH-3T3 Mausfibroblasten untersucht. Die Zellen wurden in Dulbecco'sB The influence of the substances on the growth factor-induced cell proliferation was investigated on NIH-3T3 mouse fibroblasts. The cells were in Dulbecco's
»3 modifiziertem Eagle's Medium (DMEM) mit Zusatz von 10 % fötalem Kalberserum (FKS), 2 mM Glutamin und Antibiotika/Antimykotika-Losung kultiviert Der Wechsel des Kulturmediums erfolgt regelmäßig zweimal wöchentlich Vier Tage nach der letzten Subkultivierung wurden die adherenten Zellen mit Trypsin/EDTA vom Boden der Zellkulturflasche abgelost und in einer Dichte von 50 000 Zellen pro ml in DMEM mit Zusatz von 0,5 % FKS resuspendiert Die Zellen wurden in einem Volumen von 200 μl pro Vertiefung in F-Boden Mikrotiterplatten überfuhrt und für 96 h inkubiert Nach Wechsel des Medium (DMEM ohne FKS) und Zugabe der Substanzen erfolgte 60 min spater die Restimulation der Zellproliferation durch Zugabe von 10 ng/ml rekombinantem humanem Platelet-derived Growth Factor-BB (PDGF-BB) Anschließend wurden die Zellen erneut für 24 h bei 37 °C im Brutschrank kultiviert Sechs Stunden vor der Zellernte wurde pro Testansatz 0,5 μCi Methyl-3H-Thymιdιn zugegeben Nach Ablauf der 24stundιgen Inkubationsperiode wurden die Mikrotiterplatten 5 min bei 400 g zentπfugiert und die Zelluberstande vorsichtig abpipettiert Die Zellen wurden mit Trypsin/EDTA vom Boden gelost und dann mit einem Zellharvester (Inotech) auf Glasfaser-Filter (Type G-10, ICH-201) geerntet Die Messung des Einbaues von 3H-Thymιdιn in neu synthetisierte DNA erfolge mit Hilfe eines Linear-Analyser (LB 2842, Berthold) Der Grad der anti-pro ferativen Wirkung der Substanzen wurde jeweils im Vergleich zu gleichzeitig untersuchten Losungsmittelkontrollen ermittelt»3 modified Eagle's Medium (DMEM) with the addition of 10% fetal calf serum (FCS), 2 mM glutamine and antibiotic / antifungal solution. The culture medium is changed twice a week. Four days after the last subcultivation, the adherent cells were trypsin / EDTA from The bottom of the cell culture bottle was detached and resuspended in a density of 50,000 cells per ml in DMEM with the addition of 0.5% FCS. The cells were transferred to F-bottom microtiter plates in a volume of 200 μl per well and incubated for 96 h Medium (DMEM without FCS) and the substances were added 60 minutes later, the cell proliferation was restimulated by adding 10 ng / ml of recombinant human platelet-derived growth factor-BB (PDGF-BB). The cells were then again at 37 ° for 24 h C cultivated in the incubator Six hours before the cell harvest, 0.5 μCi methyl 3 H-thyme was added per test batch after the 24-hour inku had expired During the bation period, the microtiter plates were centrifuged for 5 min at 400 g and the cell supernatants were carefully pipetted off. The cells were detached from the ground with trypsin / EDTA and then harvested with a cell harvester (Inotech) on glass fiber filter (type G-10, ICH-201). The measurement The incorporation of 3 H-thymid into newly synthesized DNA was carried out with the aid of a linear analyzer (LB 2842, Berthold). The degree of the anti-proactive effect of the substances was determined in each case in comparison to the solvent controls examined at the same time
Figure imgf000022_0001
Figure imgf000023_0001
Figure imgf000022_0001
Figure imgf000023_0001
Z\
Figure imgf000024_0001
Z \
Figure imgf000024_0001
Beispiele für die Herstellung pharmazeutischer Zubereitungen der erfindungsgemäßen Substanzen:Examples for the production of pharmaceutical preparations of the substances according to the invention:
A. Tabletten:A. tablets:
Zur Herstellung von Tabletten, die je nach gewünschter Wirkungsstärke 0.5 bis 50 mgFor the production of tablets, depending on the desired potency 0.5 to 50 mg
Wirkstoff enthalten, benötigt manContain active ingredient, you need
erfindungsgemäße Substanz 20 bis 2 000 gsubstance according to the invention 20 to 2,000 g
Zellulosepulver 2 000 gCellulose powder 2,000 g
Maisstärke 1 200 g kolloide Kieselsäure 80 gCorn starch 1 200 g colloidal silica 80 g
Magnesiumstearat 20 gMagnesium stearate 20 g
Milchzucker ad 10 000 gMilk sugar ad 10 000 g
Der Wirkstoff wird gegebenenfalls gemahlen, mit den Hilfsstoffen homogen vermischt und in der üblichen Weise zu Tabletten von je 250 mg Gewicht und 9 mm Durchmesser verpreßt. Falls gewünscht, werden die Tabletten mit einem Filmüberzug versehen.The active ingredient is optionally ground, homogeneously mixed with the excipients and compressed in the usual manner to tablets each weighing 250 mg and 9 mm in diameter. If desired, the tablets are coated with a film.
B. Kapseln:B. Capsules:
Zur Herstellung von Kapseln, die je nach gewünschter Wirkuπgsstärke 0.5 bis 50 mgFor the production of capsules, depending on the desired potency 0.5 to 50 mg
Wirkstoff enthalten, benötigt manContain active ingredient, you need
erfindungsgemäße Substanz 50 bis 5 000 gsubstance according to the invention 50 to 5,000 g
Maisstärke 2 000 g kolloide Kieselsäure 300 gCorn starch 2,000 g colloidal silica 300 g
Magnesiumstearat 50 gMagnesium stearate 50 g
Zellulosepulver ad 20 000 gCellulose powder ad 20,000 g
Die feingepulverten Stoffe werden homogen gemischt und in Hartgelatinekapseln der Größe 2 in der Menge von 200 mg pro Kapsel abgefüllt.The finely powdered substances are mixed homogeneously and filled into size 2 hard gelatin capsules in an amount of 200 mg per capsule.
U U

Claims

Patentansprüche claims
1. Zimtsäurenitrile der allgemeinen Formel I,1. cinnamic acid nitriles of the general formula I,
Figure imgf000025_0001
Figure imgf000025_0001
worin die Substituenten folgende Bedeutungen besitzen:where the substituents have the following meanings:
X = CN, COOR4, CONR5R6, COAryl, COAlkyl, Aryl, Heteroaryl,X = CN, COOR 4 , CONR 5 R 6 , COAryl, COalkyl, aryl, heteroaryl,
R1 = H, Alkyl, Arylalkyl, [4-(2-Alkoxyphenyl)-1-piperazinyl]alkyl,R 1 = H, alkyl, arylalkyl, [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl,
R2 und R3 = unabhängig voneinander H, Hydroxy, Alkoxy, Benzyl,R 2 and R 3 = independently of one another H, hydroxy, alkoxy, benzyl,
R4 = H, Alkyl, [4-(2-Alkoxyphenyl)-1 -piperazinyl]alkyl,R 4 = H, alkyl, [4- (2-alkoxyphenyl) -1 -piperazinyl] alkyl,
R5 = H, Alkyl, Alkoxyalkyl, Alkoxy-oxoalkyl, Dialkylaminoalkyl, Amino-oxoalkyl, Aryl,R 5 = H, alkyl, alkoxyalkyl, alkoxy-oxoalkyl, dialkylaminoalkyl, amino-oxoalkyl, aryl,
Arylalkyl, [4-(2-Alkoxyphenyl)-1-piperazinyl]alkyl, Morpholinoalkyl, Furanylalkyl,Arylalkyl, [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl, morpholinoalkyl, furanylalkyl,
Thienylalkyl, R6 = H, Alkyl oder NR5R6 gemeinsam = cyclisches Amin wie Pyrrolidin, Morpholin, Piperidin, subst. Piperazin, wobei mindestens einer der Reste R1 , R4 oder R5 ein [4-(2-Alkoxyphenyl)-1- piperazinyl]-alkyl-Rest ist, sowie deren Additionssalze mit pharmakologisch verträglichen Säuren.Thienylalkyl, R 6 = H, alkyl or NR 5 R 6 together = cyclic amine such as pyrrolidine, morpholine, piperidine, subst. Piperazine, at least one of the radicals R 1 , R 4 or R 5 being a [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl radical, and their addition salts with pharmacologically acceptable acids.
2. Zimtsäurenitrile nach Anspruch 1 , worin der mindestens eine [4-(2-Alkoxyphenyl)-1- piperazinyl]alkyl-Rest ein [4-(2-Methoxyphenyl)-1-piperazinyl]-propyl-Rest ist.2. Cinnamic acid nitriles according to claim 1, wherein the at least one [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl radical is a [4- (2-methoxyphenyl) -1-piperazinyl] propyl radical.
3. Arzneimittel, enthaltend mindestens eine Verbindung gemäß einem der Ansprüche 1 oder 2.3. Medicament containing at least one compound according to one of claims 1 or 2.
4. Arzneimittel nach Anspruch 3, gekennzeichnet durch eine für die Prophylaxe oder Behandlung der benignen Prostatahyperplasie (BPH) wirksame Menge an mindestens einer Verbindung gemäß einem der Ansprüche 1 bis 3 neben üblichen Hilfs- und Zusatzstoffen.4. Medicament according to claim 3, characterized by an effective for the prophylaxis or treatment of benign prostatic hyperplasia (BPH) of at least one compound according to one of claims 1 to 3 in addition to conventional auxiliaries and additives.
5. Verfahren zur Herstellung der Zimtsäurenitrile gemäß einem der Ansprüche 1 oder 2, bei dem ein Benzaldehyd der allgemeinen Formel II5. A process for the preparation of cinnamonitriles according to one of claims 1 or 2, in which a benzaldehyde of the general formula II
£3
Figure imgf000026_0001
£ 3
Figure imgf000026_0001
worin R1, R2 und R3 die in Anspruch 1 angegebenen Bedeutungen besitzen, mit einem substituierten Acetonitril der allgemeinen Formel IIIwherein R 1 , R 2 and R 3 have the meanings given in claim 1, with a substituted acetonitrile of the general formula III
Figure imgf000026_0002
Figure imgf000026_0002
worin X die in Anspruch 1 angegebenen Bedeutungen besitzt, unter Wasseraustritt zu einer Verbindung der allgem. Formel Iwherein X has the meanings given in claim 1, with water leakage to a compound of general. Formula I.
Figure imgf000026_0003
Figure imgf000026_0003
kondensiert und in an sich bekannter Weise isoliert und gereinigt wird.is condensed and isolated and purified in a manner known per se.
6. Verfahren nach Anspruch 5, dadurch gekennzeichnet, daß der Benzaldehyd und das Acetonitril in einem organischen Lösemittel gelöst werden und die Kondensation bei einer Temperatur zwischen 50°C und der Siedetemperatur während 2 Stdn. bis 3 Tagen durchgeführt wird.6. The method according to claim 5, characterized in that the benzaldehyde and the acetonitrile are dissolved in an organic solvent and the condensation is carried out at a temperature between 50 ° C and the boiling temperature for 2 hours to 3 days.
7. Verfahren nach Anspruch 5 oder 6, dadurch gekennzeichnet, daß dem Reaktionsgemisch ein cyclisches Amin als Katalysator zugegeben wird.7. The method according to claim 5 or 6, characterized in that a cyclic amine is added to the reaction mixture as a catalyst.
8. Verfahren nach einem der Ansprüche 5 bis 7, dadurch gekennzeichnet, daß das Kondensationsprodukt mit einer wäßrigen Lösung einer pharmakologisch verträglichen Säure versetzt und das gebildete Säureadditionssalz abgetrennt wird.8. The method according to any one of claims 5 to 7, characterized in that the condensation product is mixed with an aqueous solution of a pharmacologically acceptable acid and the acid addition salt formed is separated off.
9. Reaktive Zwischenprodukte der allgemeinen Formel II9. Reactive intermediates of the general formula II
m
Figure imgf000027_0001
m
Figure imgf000027_0001
worinwherein
R )1 = [4-(2-Alkoxyphenyl)-1-piperazinyl]alkylR) 1 = [4- (2-alkoxyphenyl) -1-piperazinyl] alkyl
R2 und R3 = unabhängig voneinander H, Hydroxy, Alkoxy mit 1 - 4, vorzugsweise mit 1 - 2R 2 and R 3 = independently of one another H, hydroxy, alkoxy with 1-4, preferably with 1-2
C-Atomen, Benzyl, bedeuten.C atoms, benzyl.
t5 t5
PCT/EP2000/008419 1999-09-01 2000-08-29 Cinnamic acid nitrile method of production and use thereof WO2001016124A1 (en)

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US4689327A (en) * 1984-06-30 1987-08-25 Tanabe Seiyaku Co., Ltd. N-substituted-2-[2-[2-(4-phenylpiperazine-1-yl)ethoxy]phenyl]-thiazolidine-3-carboxamides useful as cardiotonic agent
EP0558245A1 (en) * 1992-02-25 1993-09-01 RECORDATI S.A. CHEMICAL and PHARMACEUTICAL COMPANY Heterobicyclic compounds as antagogists of alpha-1 adrenergic and SHT1A receptors
JPH08143557A (en) * 1994-11-24 1996-06-04 Toyobo Co Ltd New phenylpiperazine derivative, its salt and its solvated material
JPH0912563A (en) * 1995-06-30 1997-01-14 Toyobo Co Ltd New benzylalcohol derivative, its production and its use

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US4689327A (en) * 1984-06-30 1987-08-25 Tanabe Seiyaku Co., Ltd. N-substituted-2-[2-[2-(4-phenylpiperazine-1-yl)ethoxy]phenyl]-thiazolidine-3-carboxamides useful as cardiotonic agent
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