Derivatives of pyrimidine-2,4,6-trione and pharmaceutical composition containing these compounds

FIELD: organic chemistry, biochemistry, pharmacy.

SUBSTANCE: invention relates to new compounds of the formula (I):

eliciting inhibitory activity with respect to metalloproteinases and wherein R1 means phenoxy-group wherein phenyl residue can be substituted with one or some halogen atoms, hydroxy-, (C1-C6)-alkoxy-group, (C1-C6)-alkyl, cyano- or nitro-group; R2 means pyrimidine, pyrazine or its N-oxide or phenyl substituted with -SO2NR3R4 wherein R3 and R4 can be similar or different and mean hydrogen atom, direct-chain or branch-chain (C1-C6)-alkyl that can be substituted once or some times with the group OH, N(CH3)2, or it can be broken by oxygen atom, or it represents COR5 wherein R5 means (C1-C)-alkyl group that can be substituted with NH2. Also, invention relates to a pharmaceutical composition comprising above said compounds.

EFFECT: valuable biochemical properties of compounds and composition.

5 cl, 1 sch, 1 tbl, 10 ex

 

The present invention relates to new derivatives of 5,5-disubstituted pyrimidine-2,4,6-Trion. These compounds display a marked antitumor and antimetastatic activity.

In normal tissues, there is an equilibrium between synthesis and degradation. Degradation of the extracellular matrix occurs under the action of proteases that belong to at least three groups of extracellular metalloproteinases. These include collagenase, gelatinase and stromelysin. In the normal state there are specific inhibitors for these catabolic enzymes, such as α2-macroglobulin and TIMP [tissue inhibitor of metalloproteinases (MMP)], so excessive degradation of the extracellular matrix occurs. To the group of proteases are Adamantine.

The most famous member of the group agamaliev is TACE (TNF-α -converting enzyme). It was characterized by at least 17 different and also highly homologous species DFID, including interstitial fibroblast collagenase (MMP-1, HFC), neutrophil collagenase (MMP-8, HNC), two gelatinase, stromelysin, such as the HSL-1, and HPUMP (see the recent review Birkedal-Hansen, H., Moore, W.G.I., Bodden, M.K., Windsor, L.J., Birkedal-Hansen, B., DeCarlo, A., Engler, J.A., Critical Rev.Oral Biol.Med. (1993) 4, 197-250). These proteases have a number of common structural and functional features, but there are a few who by their substrate specificity. Only HNC and HFC able to split one connection native transferline the collagens type I, II and III with the formation of fragments with a length of 3/4 and 1/4 native circuit. It lowers the melting temperature of collagen and makes it available for further attack by other enzymes, degrading the extracellular matrix.

However, uncontrolled excessive degradation of this extracellular matrix characteristic of many pathological conditions, such as, for example, manifested in the clinic as rheumatoid arthritis, osteoarthritis and multiple sclerosis, the formation of tumor metastasis, corneal ulcer, inflammatory disease and infestation, disease of bones and teeth.

However, it can be assumed that the pathogenesis of this clinical picture can favorably be influenced by the introduction of matrix metalloproteinases inhibitors of extracellular matrix. Meanwhile, from the scientific literature [see, for example, a review article D.E. Levy, A.M. Ezrin in Emerging Drugs 2, 205-230 (1997), M. Whittaker, P. Brown, Curr.Opin.Drug Discovery Dev. (1998), 1(2), 157-164] known or described in the patent literature a number of compounds, mainly with the remainder of hydroxamic acids, thiol or phosphine group as connecting the zinc group (see, for example, among other applications WO-A 9209563 filed by the company Glycomed, EP-A 497192 filed by the company Hoffmann-LaRoche, WO-A 9005719 filed by the company British Biotechnology, EP-A 489577 filed by the company Celltech, EP-A 320118, the odata firm Beecham, patent US 4595700 issued by the company Searle application WO 97/20824 filed by the company Agouron Pharmaceuticals, WO 96/15096 filed by the company Voeg Corporation).

Some of these compounds are active as inhibitors of extracellular proteases, but their suitability for oral use is extremely low. Such compounds often have a broad spectrum inhibition of metalloproteinases, which may cause undesirable side effects and toxicity.

Derivatives of pyrimidine-2,4,6-Trion have been described in EP 0869947 mainly as inhibitors of extracellular metalloproteinases. However, there is a high need for new compounds with low toxicity, does not exhibit side effects and showing significant inhibitory activity against metalloproteinases, in particular the need for inhibitors as candidates for the treatment of chronic diseases, in particular tumor growth and metastasis.

It was found that in comparison with the compounds claimed in EP 0869947 proposed new derivatives of pyrimidine-2,4,6-trione have increased activity as inhibitors of extracellular proteases, and in addition acceptable for oral administration.

The present invention relates to compounds of General formula I

in which R1denotes phenyl, phenoxy,phenylthio-, phenylsulfinyl, phenylsulfonyl, phenylamino or phenylmethylene group in which the phenyl residue may be substituted by one or more halogen atoms, hydroxy, C1-C6alkoxy, C1-C6alkyl, cyano - or nitro groups, and the preferred substitution by one or two substituents in the para and/or meta-position;

R2denotes optionally substituted aryl or retailnow group.

The present invention also includes pharmaceutically acceptable salts and prodrugs based on compounds of the formula I, as well as the use of these compounds in the preparation of pharmaceutical medicines.

Aryl group specified among the values of R2consists of the phenyl ring. Under retailnow group understand the cyclic unsaturated or saturated ring system containing from 5 to 7 ring atoms, which may be selected from one or more atoms of carbon, nitrogen, oxygen or sulfur. Preferred getaline radicals with a lack of electrons, such as nitrogen-containing 6-membered rings like pyridine, pyrimidine, pyrazinium or 1,3,5-triazine or N-oxides. The most preferred getrolename radicals are pyrimidinyl or pyrazinyl. Aryl or getaline rings can be replaced by the ne or more substituents, selected from a halogen atom, hydroxyl, alkoxy, amino, dialkylamino-, cyano groups, (ness.)of alkyl, respectively (ness.)of alkyl, (ness.)alkenyl, (ness.)the quinil, (ness.)acyl, (ness.)alkylthio, (ness.)alkylsulfonyl, (ness.)alkylaminocarbonyl, aminocarbonyl, SO2NR3R4, nitro, (ness.)alkoxycarbonyl, carboxyl, where each of R3and R4that may be the same or different, denotes a hydrogen atom, remotemachine or branched C1-C6alkyl which may be substituted one or more times by a group of HE, N(CH3)2or which can be interrupted by oxygen atom, or represents a R5where R5denotes an alkyl group which may be substituted NH2. The preferred substitution of one or two of the above substituents in the para and/or meta-position.

The term "(ness.)alkyl" in the radical R2as such or in combination with other radicals, denotes C1-C6alkyl, preferably methyl, ethyl, propyl, isopropyl or tert-butyl.

The term "(ness.) alkenyl" refers to C2-C6alkenyl, preferably allyl or pentadienyl. The term "(ness.)quinil" refers to C2-C6quinil, preferably propargyl.

In all cases under (ness.)the acyl in the radical R2units is meaut-C(O)-C 1-C6alkyl or-C(O)H, preferably acetyl group.

Alkyl residues R2can, but not necessarily, be interrupted once or more heteroatoms (O, S, NH).

Under the halogen atom understand the atom of fluorine, chlorine, bromine or iodine, preferably chlorine or bromine.

If compounds of General formula I contain one or more asymmetric carbon atoms, an object of the present invention are optically active compounds of General formula I.

Compounds of General formula I can be synthesized by well known methods, and in the preferred embodiment, carried out the reaction of compounds of General formula II

in which R1have the above values, and T denotes a leaving group, such as Hal or OSO2R3. Hal denotes a chlorine atom, bromine or iodine, and R3denotes aryl or methyl group, with compounds of General formula III

in which R2has the values listed above, with an optional transformation into a pharmaceutically acceptable salt.

Compounds of General formula II can be synthesized by methods similar to known from the literature. So, for example, pyrimidine-2,4,6-Trion, brominated at the 5-position can be synthesized by the reaction of suitable dialkylamino esters of bromomalonate acid with urea [see, for example, Acta Chim.Acad.Sci.Hung. 107 (2), 139 (1981)]. The corresponding brominated or chlorinated compounds of General formula II can be obtained by the reaction of a pyrimidine-2,4,6-Trion, substituted R1the phenyl in the 5th position with bromine [similar to that presented in J.Prakt.Chemie 136, 329 (1933) or J.Chem.Soc. 1931, 1870] or sulfurylchloride (see J.Chem.Soc. 1938, 1622), or N-bromosuccinimide, or the same romirowsky agents. Such methods are also described in EP 0869947.

Amines of General formula III is technically available or generally known from the literature, or equivalent, used in the methods described in the experimental part.

Amines of General formula III, where R2pyrazin or pyrimidine N-oxide are disclosed, for example, in U.S. patents 4 082 844, USA 4 081 542 and Catto. A. et. al. Boll. Chim. Farm. 121(1982) 15-26.

Pyrimidine-2,4,6-Trion formula II in which T represents a hydrogen atom, can be obtained in accordance with known methods by reaction of esters of malonic acid with urea [see, e.g., J.Med.Chem. 10, 1078 (1967) or Helvetica Chim.Acta 34, 459 (1959), Pharmacie 38 (1), 65 (1983), or EP 0869947]. These reactions are usually carried out in alcohols, such as methanol, ethanol or butanol, in the presence of the corresponding sodium alcoholate at a temperature in the range from 40 to 100°C.

The esters of malonic acid, which is necessary to provide a pyrimidine-2,4,6-Trion known from the literature or can be obtained in accordance is known from the literature methods. The usual method of obtaining derivatives of malonic acid, in which R1have the above values, is illustrated in the following diagram:

Examples of such reactions can be found in Houben-Weyl volume E5/2 in J. Org. Chem. 46, 2999 (1981) and Arch. Pharm. 323, 579 (1990).

Compounds of General formula I can contain one or more chiral centers and in these cases can be in racemic or in optically active form. In accordance with known methods of the racemates can be separated into the enantiomers. Preferred diastereoisomeric salts which can be separated by crystallization, obtained from racemic mixtures by reaction with an optically active acid, such as, for example, D-or L-tartaric acid, mandelic acid, malic acid, lactic acid or camphorsulfonic acid, or optically active amine, such as, for example, D - or L-α -phenylethylamine, ephedrine, quinidine or cinchonidine. As pharmaceutically acceptable salts are used mainly alkaline salts, alkaline earth metal, similar to the salts of CA or Mg, ammonium salts, acetates or hydrochloride who receive the usual way, for example by titration of inorganic compounds or organic bases or mineral acids such as, for example, sodium hydroxide, potassium hydroxide, aqueous ammonia, 1-C4the bonds alkylamines, such as, for example, triethylamine or hydrochloric acid. Typically, such salts are purified by precipitation from water/acetone.

Proposed in accordance with the invention the new compounds of formula I and their salts can be introduced into the body interline or parenterally in liquid or solid form. In this regard, take into account all of the usual form of introduction into the organism, such as, for example, tablets, capsules, coated tablets, syrups, solutions, suspensions, etc. In a preferred embodiment, in the form of medium for injection use water which contains the additives, such as stabilizers, soljubilizatory and buffers, which is normal in solutions for injection.

Such additives are, for example, tartrate and citrate buffers, ethanol, complex-forming agents (such as ethylenediaminetetraacetic acid and its non-toxic salt), high-molecular polymers (such as liquid polyethylene oxide) for regulating the viscosity. Liquid substances-carriers for the preparation of solutions for injection should be sterile, therefore, in a preferred variant, they stariway in ampoules. Solid carriers are, for example, starch, lactose, mannitol, methylcellulose, talc, highly dispersed silicic acid, high molecular weight fatty acids (such as stearic acid), gelatin, agar-agar, F. the SFAT calcium, magnesium stearate, animal and vegetable fats, solid high molecular weight polymers (such as polyethylene glycols). Acceptable for oral administration, the preparations may also, but not necessarily, include corrigentov and sweeteners.

The dose depends on various factors, such as the introduction into the organism, a species of the patient, age and/or individual state of health. It is necessary to incorporate a daily dose is from about 10 to 1000 mg for humans, preferably from 100 to 500 mg for a person, and you can enter them once in the form of a dose at one time or several times in fractional doses.

Prodrugs are compounds according to the invention, which in vivo are converted into pharmacologically active compounds. The most common prodrugs are esters of carboxylic acids.

Example 1

5-[4-(4-chlorophenoxy)phenyl]-5-(4-(pyrimidine-2-reperation)pyrimidine-2,4,6-Trion

A) 1-[4-(4-chlorophenoxy)phenyl]alanon

24.4 g of 4-fortetienne dissolved in 180 ml of dimethylformamide and added to 22.8 g of 4-chlorophenol and 29.5 g of potassium carbonate. The mixture is refluxed with stirring for 7 hours After cooling, the mixture is diluted with water and extracted with methylene chloride. The organic phase is washed with water, dried and evaporated to obtain 38 g of crystalline TV is Gogo substances, tPL: 66-68° C.

B) 2-[4-(4-chlorophenoxy)phenyl]morpholine-4-retention

12.4 g of the product obtained according to the stages outlined above, is mixed with 4 g of sulfur and 8.8 ml of the research. The mixture was kept at 150°C for 2 h, cooled in an ice bath and treated for 30 min with 20 ml of ethanol. Precipitated crystals are collected and recrystallized from ethanol to obtain 13 g specified in the title compounds. tPL: 104-105° C.

C) [4-(4-chlorophenoxy)phenyl]acetic acid

10.4 g of the compound obtained in stage B, incubated with 200 ml of 50%sulfuric acid at 130° within 8 hours After cooling to room temperature the reaction mixture is diluted with 300 ml of water and extracted with ethyl acetate. The organic phase is washed with water and further extracted with 2 N. solution of sodium carbonate. The aqueous phase is acidified with diluted hydrochloric acid, add ethyl acetate, the organic phase is separated, dried and evaporated to obtain a 5.1 g of a brownish residue, tPL: 98-100° C.

D) methyl ester [4-(4-chlorophenoxy)phenyl]acetic acid

to 5.1 g of the product from stage dissolved in 50 ml of methanol. The solution is cooled to -10° and treated with 3 ml of thionyl chloride, and then refluxed for 1 h, the Reaction mixture is evaporated and the residue is dissolved in diethyl ether. Efiro the phase washed with water, dried and evaporated to obtain a 5.1 g of the product as a reddish brown oil.

D) dimethyl ester 2-[4-(4-chlorophenoxy)phenyl]malonic acid

A suspension of 350 mg of sodium hydride in 10 ml of dimethylcarbonate at room temperature is treated with the product obtained at stage, the Mixture is maintained at 90° C for 1 h, cooled, poured into ice water and extracted with methylene chloride. The extract is dried and evaporated to produce in the form of oil 5.7 g specified in the title compound.

E) 5-[4-(4-chlorophenoxy)phenyl]pyrimidine-2,4,6-Trion

800 mg of sodium are dissolved in 80 ml of ethanol. To this solution was added 1.65 g of urea and a solution of 5.5 g obtained by the above compounds in ethanol. The mixture is refluxed for 3 h, cooled to room temperature, treated with 100 ml of ice water and acidified with diluted hydrochloric acid. The precipitate is collected, washed with water and dried to obtain 5 g specified in the title compounds. tPL: 257-258° C.

G) 5-bromo-5-[4-(4-chlorophenoxy)phenyl]pyrimidine-2,4,6-Trion

A suspension of 6.3 g of compound obtained in stage E, 4.1 g of N-bromosuccinimide and 100 mg dibenzoylperoxide in 120 ml of carbon tetrachloride is stirred for 3 h at room temperature. The mixture is evaporated and the residue extracted with ethyl acetate. The organic phase is dried and evaporated to obtain is in the form of a thick oil 7.5 g specified in the title compound.

C) 5-[4-(4-chlorophenoxy)phenyl]-5-(4-(pyrimidine-2-reperation)pyrimidine-2,4,6-Trion

A solution of 410 mg of the compound from step G in 5 ml of methanol is treated with 330 mg of N-(pyrimidine-2-yl)piperazine. The mixture is stirred for 24 hours Obtained after evaporation of the reaction mixture the residue chromatographic on silica gel using methylene chloride/5% methanol as eluent. By combining the relevant fractions in the form of amorphous solids obtain 410 mg specified in the title compound, which identify mass spectroscopy: m/e 492.

Example 2

5-[4-(4-chlorophenoxy)phenyl]-5-(2,3,5,6-tetrahydro[1,2']bipyridinyl-4-yl)pyrimidine-2,4,6-Trion

Specified in the title compound get similar stage C of example 1 using 330 mg of 1-(pyrazin-2-yl)piperazine instead of N-(pyrimidine-2-yl)piperazine with output in the form of amorphous solids 460 mg mentioned in the title compound, which identify mass spectrometry: m/e 492.

Example 3

Analogously to example 1 by replacing 4-chlorophenol corresponding phenols obtain the following compounds. The final products identify mass spectrometry.

Example 4

4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}-N-(2-hydroxyethyl)benzosulfimide

A) N-(2-hydroxyethyl)-4-Pieper is Zin-1-albenzaalbenza

4-forbindelsesfaneblad dissolved in 20 ml of dichloromethane and treated with a solution of 1.2 ml of ethanolamine in 10 ml of dichloromethane. The mixture is stirred for 1 h and extracted twice with 50 ml water. The aqueous phase is saturated with sodium chloride and extracted twice with ethyl acetate. The combined organic phases are dried with magnesium sulfate and evaporated. 1.4 g of the obtained 4-fluoro-N-gidroxiatilenzellulozu dissolved in 15 ml of water and treated with 2.6 g of piperazine. The mixture is refluxed for 6 h and incubated at room temperature for 24 hours, the Residue is collected, washed with a small amount of water and dried to obtain 1.6 g specified in the title compound, which identify mass spectrometry (APCI [M+H]=286).

B) 4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}-N-(2-hydroxyethyl)benzosulfimide

A solution of 230 mg of the compound from step G of example 1 in 5 ml of methanol is treated with 330 mg of N-(2-hydroxyethyl)-4-piperazine-1-albenzaalbenza (see above). The mixture is stirred for 24 hours of the Residue obtained after evaporation of the reaction mixture, chromatographic on silica gel using methylene chloride/15% methanol as eluent. By combining the relevant fractions in the form of amorphous solids obtain 186 mg specified in the title compound, to the which identify mass spectrometry: APCI[M+1]=614.

Example 5

Using the methods of example 1, replacing, when necessary, 4-chlorophenol corresponding phenols obtain the following compounds. Data pieperazinove derivatives gain in accordance with stage a of example 4 and substitution of ethanolamine appropriate amine. The final products identify mass spectrometry.

Example 6

N-(2-oxo[1,3]dioxolane-4-ylmethyl)-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide

120 mg product No. 11 of example 5 dissolved in a mixture of 5 ml dichloromethane and 5 ml of tetrahydrofuran. The solution is treated with 65 mg of N,N'-carbonyldiimidazole and stirred at room temperature for 4 hours the Solvent is evaporated and the residue chromatographic on silica gel using dichloromethane/methanol in the ratio 9:1 as the eluting solvent. In the evaporation contains the product fractions are given 60 mg specified in the title compound, mass-spectrogram: APCI[M+H]=634, [M-N=636.

Example 7

N-(4-aminobutyryl)-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide

A) 4-(4-benzylpiperazine-1-yl)benzosulfimide

25 g of 4-forbindelsesfaneblad dissolved in 250 ml of di is loretana and treated at 0° With 50 ml of 25%aqueous ammonia solution. The mixture is stirred for 2 h at low temperature and overnight at room temperature. The reaction mixture is acidified and evaporated organic solvent. The residue is extracted with ethyl acetate to obtain 20 g of 4-forbindelsesfaneblad which is dissolved in 300 ml of water, treated with 102 g of 1-benzylpiperazine and refluxed for 24 hours, the Reaction mixture is filtered to obtain 26 g specified in the title compound (mass spectrogram: APCI [M+H]=332).

B) tert-butyl ether 4-[4-(piperazine-1-yl)benzosulfimide]-4-oxobutanamide acid

of 3.05 g of 4-(N-tert-butoxycarbonyl)aminobutyric acid dissolved in 30 ml of tetrahydrofuran and treated with 2.5 g of N,N'-carbonyldiimidazole.

The mixture is stirred at room temperature for 15 min, refluxed for 15 min and stirred for 1 h at room temperature. Added to 3.3 g of the product from step a and the mixture is stirred over night. The solvent is evaporated and the residue is mixed with dichloromethane and water. The organic phase is separated, dried and the solvent is evaporated. The remainder chromatographic on silica gel using dichloromethane/methanol in the ratio 9:1 as the eluting solvent. The product is subjected to catalytic hydrogen the emission in methanol, using Pd on charcoal, to obtain 2.5 g specified in the title compound (mass spectrogram: APCI[M-N]=425).

C) N-(4-aminobutyryl)-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide

Similarly to the method of stage 3 of example 1, the product obtained at stage B, enter into an interaction with 5-bromo-5-[4-(phenoxy)phenyl]pyrimidine-2,4,6-Trion. This is the last connection receive similarly to the methods described in example 1, substituting p-chlorophenol and phenol. To remove the protecting SIDE (tert-butoxycarbonyl) group 290 mg of the product is dissolved in 4 BC Hcl solution in hexane. After 1 h of aging at room temperature, the solution is decanted and the residue is ground to powder with diethyl ether, receiving 180 mg specified in the title compound (mass spectrogram: APCI[M+H]=621).

Example 8

Using the methods of example 7 with the substitution of 4-(N-tert-butoxycarbonyl)aminobutyric acid corresponding protected N-tert-butoxycarbonyl amino acids have the following connections. The final products identify mass spectrometry.

No.Chemical nameThe results of MS APCI [M+H]
1N-aminoacetyl-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide 593
2N-(5-aminopentanoic)-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide635

3N-(5-aminopentanoic)-4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}benzosulfimide669
4N-(4-aminobutyryl)-4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl} benzosulfimide655

Example 9

4-methoxybenzyloxy ester 2-oxo-2-{4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide}ethyl) - carbamino acid

140 mg of product No. 1 of example 5 is dissolved in 10 ml of dichloromethane, mixed with of 0.14 ml of triethylamine and treated with 4-methoxyphenylacetamide. The mixture is stirred for 90 min at room temperature and evaporated. The remainder chromatographic on silica gel using dichloromethane/methanol (ratio 9:1) as eluent. By combining the appropriate fractions receive 90 mg specified in the title compound (mass spectrogram: APCI[M+H]=743).

Example 10

To determine the inhibition of MMP, in particular HNC (MMP-8), catalytic domain [isolation and purification, see, e.g. the, in the work Schnierer, S., Kleine T., Gote, T., Hillemann A., Knauper V, Tschesche H., Biochem. Biophys. Res.Commun. (1993) 191, 319-326] were incubated with inhibitors, taken at various concentrations. Then according to the method similar to that described Grams F. and others [FEBS 335 (1993) 76-80], determined the initial reaction rate for the conversion of standard substrates.

The results were evaluated by the graphical dependence of the inverse of the reaction rate on the concentration of the inhibitor. The inhibition constant (Ki) was determined by a graphical method M. Dixon, Biochem. J. (1953) 55, 170-202 as a negative piece on the x-axis.

Synthetic substrate for collagenase was heptapeptide, With the end of which was chemically attached group DNP (dinitrophenol). Due to steric hindrance specified DNF residue quench the fluorescence of the neighboring residue of tryptophan in heptapeptide. After splitting Tripeptide containing DNP group, the fluorescence of tryptophan increases. Thus, proteolytic cleavage of the substrate can be determined by the value of the fluorescence.

a) the First method

The analysis was carried out at 25° in freshly prepared 50 mm Tris-buffer (pH 8.0)to remove heavy metals processed by dithizone. Added 4 mm CaCl2and the buffer was saturated with argon. Alternate solutions Adamantine II were prepared by centrifugation of the protein suspension in ammonium sulfate and after the ith its dissolution in the buffer for analysis. Alternate solutions collagenase was diluted with buffer for analysis. The concentration of the enzyme was determined by absorption in ultraviolet rays (ε280=2,8· 10-4M-1·cm-1that ε288=2,2· 10-4M-1·cm-1and spare the solution was kept cold. This solution was diluted 100-fold to a final concentration of 16 nm. Fluorogenic substrate DNP-Pro-Leu-Gly-Leu-Trp-Ala-D-Arg-NH2with Km52 μm was used at a concentration of 21,4 μm; To determineialso used a solution with a concentration of 12.8 μm. The fluorescence of the substrate was determined at wavelengths of excitation and emission, respectively λ =320 and 420 nm on an (Perkin Elmer Model 650-40), holder equipped with a thermostatted cell. For the hydrolysis of the substrate was observed within 10 min immediately after adding the enzyme. All experiments were performed at least three repetitions. Values Forifor the inhibitors was calculated by the point of intersection of the straight lines obtained from the graphical dependencies vo/vithe concentration of inhibitor, while values IR50expected from graphic dependences vi/vothe concentration of inhibitor which data were processed by the method of nonlinear regression with simple normal weighing.

b) the Second method

The buffer on the I analysis:

50 mm Tris/Hcl with a pH of 7.6 [Tris refers to Tris(hydroxymethyl)aminomethane]

100 mm NaCl/10 mm CaCl2/5% MeOH (if necessary)

Enzyme: 8 nm catalytic domain (Met80-Gly242) collagenase human neutrophils (MMP-8)

Substrate: 10 μm DNP-Pro-Leu-Gly-Leu-Trp-Ala-D-Arg-NH2

Total volume for analysis: 1 ml

The solution of enzyme and inhibitor were prepared in buffer for analysis (25°). The reaction was initiated by introducing into the substrate solution. For splitting fluorogenic substrate was monitored using fluorescence spectroscopy, using the wavelength of excitation and emission, respectively 280 and 350 nm. Values IR50was calculated as the concentration of inhibitor required to reduce the reaction rate by half in comparison with the rate of the reaction without inhibitor.

Table 1 shows the values IR50in comparison with the same values for the compounds of example 26 and the preferred connection No. 118, cited in EP 0869947.

Table 1

Values IR50for MMP inhibitor (anti-MMP-8 catalytic domain)
Connection EP 0869947IR50[nm]
preferred No. 11860
Example 2615
Connections really hard is retenu IR50[nm]
Example 110
Example 24
Example 3-No. 14
Example 3-No. 22
Example 3-No. 154
Example 3-No. 154
Example 410
Example 5-No. 62,8
Example 5-No. 713
Example 5-No. 912
Example 5-No. 109
Example 5-No. 114,5
Example 5-number 125,5
Example 5-No. 136
Example 5-number 1813
Example 5-No. 199
Example 69

1. The compounds of formula

in which

R1denotes fenoxaprop, in which the phenyl residue may be substituted by one or more halogen atoms, hydroxy, C1-C6alkoxy, C1-C6alkyl, cyano - or nitro group, and

R2denotes a pyrimidine, pyrazin or its N-oxide or phenyl, substituted-SO2NR3R4,

where R3and R4that may be the same or different and, represent a hydrogen atom, remotemachine or branched C1-C6alkyl which may be substituted one or more times by a group of HE, N(CH3)2or which can be interrupted by oxygen atom, or represents a COR5where R5represents C1-C4is an alkyl group which may be substituted NH2.

2. Compounds according to claim 1 of formula I,

in which R1denotes peroxyradical, substituted one or more times by chlorine atoms, bromine, stands or tert-bootrom.

3. Compounds according to claim 1 of formula I, in which R3denotes a hydrogen atom, and R4denotes a hydrogen atom, -CH2CH2HE; -CH2CH2-N(CH3)2; -CH2-CH(OH)-CH2OH; -CH-(CH2OH)2; -CH2-CH2-O-CH2CH2-O-CH2CH2HE or-C(CH2OH)3.

4. Compounds according to claim 1 of formula I, selected from a range including:

5-[4-(4-chlorophenoxy)phenyl]-5-(4-(pyrimidine-2-reperation)pyrimidine-2,4,6-Trion,

5-[4-(4-chlorophenoxy)phenyl]-5-(2,3,5,6-tetrahydro[1,2']bipyridinyl-4-yl)pyrimidine-2,4,6-Trion,

5-[4-(3,4-dichlorophenoxy)phenyl]-5-(4-(pyrimidine-2-reparation-1-yl)pyrimidine-2,4,6-Trion,

5-[4-(3,4-dichlorophenoxy)phenyl]-5-(2,3,5,6-tetrahydro[1,2']bipyridinyl-4-yl)pyrimidine-2,4,6-Trion,

5-[4-(4-bromophenoxy)phenyl]-5-(4-(pyrimidine-2-reparation--yl)pyrimidine-2,4,6-Trion,

4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}-N-(2-hydroxyethyl)benzosulfimide,

4-{4-(5-[4-(4-bromophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}benzosulfimide,

4-{4-(5-[4-(4-bromophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}-N-(2-dimethylaminoethyl)benzosulfimide,

4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}benzosulfimide,

4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}-N,N-bis(2-hydroxyethyl)benzosulfimide,

N-(2,3-dihydroxypropyl)-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide,

N-(2-hydroxy-1-hydroxymethylene)-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide,

N-2-[2-(2-hydroxyethoxy)ethoxy]ethyl-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide,

N-(2-hydroxy-1,1-bishydroxyethyl)-4-(4-[2,4,6-trioxo-5-(4-phenoxyphenyl)hexahydropirimidine series-5-yl]piperazine-1-yl)benzosulfimide,

4-{4-(5-[4-(4-chlorophenoxy)phenyl]-2,4,6-trioxalatoferrate-5-yl)piperazine-1-yl}-N-(2-hydroxy-1,1-bishydroxyethyl)benzosulfimide.

5. The pharmaceutical composition exhibiting and guruuuu activity against metalloproteinases, including a connection according to claims 1 to 4 in a mixture with pharmaceutically acceptable excipients or diluents.



 

Same patents:

FIELD: organic chemistry, heterocyclic compounds, medicine, pharmacy.

SUBSTANCE: invention relates to nitrogen-containing heterocyclic derivatives of the formula (I): A-B-D-E (I) wherein A means 5- or 6-membered heteroaryl comprising one or two nitrogen atoms in ring; B means ethenylene; D mean phenylene; E means group -N(COR)-SO2-G wherein G means phenyl; R means 5- or 6-membered heteroaryl or heteroarylmethyl comprising one or two nitrogen atoms in ring, or group -(CH2)n-N(R5)R6 wherein n means a whole number from 1 to 5; R5 and R6 are similar or different and mean: hydrogen atom, (C1-C6)-alkyl, hydroxyalkyl, aminoalkyl; or R5 and R6 in common with nitrogen atom can form 5-7-membered cyclic amino-group -N(R5)R6 that can comprise, except for nitrogen atom, also oxygen, sulfur or nitrogen atom as a component forming the ring, or their N-oxides. Compounds of the formula (I) elicit anticancer activity and can be used in medicine.

EFFECT: valuable medicinal properties of compounds.

10 cl, 1 tbl, 24 ex

FIELD: organic chemistry, chemical technology, medicine, pharmacy.

SUBSTANCE: invention relates to new derivatives of piperazinylalkylthiopyrimidine of the formula (I): wherein R1 represents hydrogen atom, (C1-C4)-alkyl, (C1-C4)-alkanoyl or di-(C1-C4-alkyl)-amino-(C1-C4-alkyl); R2 means hydrogen atom or benzyl substituted with 1-3 substitutes taken among the group consisting of (C1-C4)-alkyl, (C1-C4)-alkoxy-group, di-(C1-C4-alkyl)-amino-group, hydroxyl group and halogen atom; n = 2, 3 or 4, and to its pharmaceutically acceptable acid addition salt. Also, invention describes a method for preparing compounds and pharmaceutical composition based on thereof. Compounds are useful for treatment of diseases arising as result of the central nervous system injury.

EFFECT: improved preparing method, valuable medicinal properties of compounds.

14 cl, 3 tbl, 26 ex

The invention relates to organic chemistry and can find application in medicine

The invention relates to new nitrogen-containing aromatic 6-membered cyclic compounds of the formula (I) or their pharmaceutically acceptable salts, demonstrating excellent selective PDE V inhibitory activity

Thrombin inhibitors // 2221808
The invention relates to compounds of formula I, the values of the radicals defined in the claims and their pharmaceutically acceptable salts

The invention relates to substituted cyclic aminoven compounds of formula (I)

< / BR>
where Ar represents thienyl, substituted pyridine, phenyl unsubstituted or substituted with halogen, hydroxy, alkoxy, C1-C4the alkyl, phenyloxy, NO2or phenyl; R1is NHOR2where R2is hydrogen; W is one or more hydrogen atoms; Y is independently one or more members of the group consisting of hydroxy, SR3, alkoxy, NR6R7where R6and R7independently selected from hydrogen, alkyl, pyridylethyl, SO2R8, COR9or R6and R7can be combined with the formation of the ring containing the nitrogen to which they relate, formulas

< / BR>
where Y' is CH2OH , SO2; R3represents hydrogen, alkyl, aryl, benzothiazolyl, pyrazinyl, N-methylimidazole; R8represents C1-C4alkyl, phenyl; R9represents hydrogen, alkyl, phenyl; Z is hydrogen; n = 1, and its optical isomer, diastereoisomer, or enantiomer, or its pharmaceutically acceptable salt

The invention relates to compounds of formula (I)

< / BR>
in which Ar1denotes a heterocyclic group, which represents a pyrazole which may be substituted by one or more radicals R1, R2or R3; Ar2denotes phenyl, naphthyl or tetrahydronaphthyl, each of which optionally is substituted by one to three groups R2; L denotes a saturated or unsaturated, branched or unbranched carbon C1-C10chain; in which one or more methylene groups are optionally independently replaced by O, NH or S, and in which the linking group is optionally substituted by 0-2 of doxography; Q has a value selected from a range of: a) phenyl, naphthyl, pyridine, imidazole, Piran, etc. b) tetrahydropyran, morpholine, thiomorpholine, thiomorpholine and t

The invention relates to new derivatives of benzothiadiazole, benzoxazoles and benzodiazines formula I in free base form or in the form of a pharmaceutically acceptable acid salt additive that can be used as an anxiolytic drug in the treatment of any condition, which is associated with increased endogenous levels of CRF or in which violated the regulation of the hPa system (hypothalamic - pituitary), or various diseases that are caused by CRF1or the manifestation of which contributes CRF1such as arthritis, asthma, allergies, anxiety, depression, etc

The invention relates to amide derivative of the formula I

< / BR>
where R3represents (1-6C)alkyl or halogen; m is 0, 1, 2 or 3; R1represents hydroxy, halogen, trifluoromethyl, nitro, amino, (1-6C)alkyl, (2-6C)alkenyl, (2-6C)quinil, (1-6C)alkoxy, (1-6C)alkylamino, di-[(1-6C)alkyl] amino, amino-(2-6C)alkylamino, (1-6C)alkylamino-(2-6C)alkylamino etc

The invention relates to new imidazole compounds of the formula I:

where R1represents hydrogen, hydroxy, protected hydroxy, or aryl, optionally substituted with a suitable(and) substituent(s) selected from the group consisting of halogen(lower)alkyl, halogen, hydroxy, protected carboxy, carbamoyl, lower alkylenedioxy, lower alkoxy, optionally substituted aryl, and lower alkyl, optionally substituted by hydroxy or protected carboxy; R2represents hydrogen or lower alkyl; R3is hydroxy or protected hydroxy; R4represents cyano, (hydroxy)minamino(lower)alkyl, carboxy, protected carboxy, N-containing heterocyclic group, optionally substituted amino, or carbarnoyl, optionally substituted with a suitable(s) of the substituent(s) selected from the group consisting of amino, hydroxy, lower alkyl, lower alkylsulfonyl, amidoamine(lower)alkyl, optionally substituted by hydroxy; and-And - is-Q -, or-O-Q-, where Q is a single bond or lower alkylene, or its salt, provided when R2is the lowest Ala the substituent(s), the above, and also provided that the compound of formula I is not 1-(hydroxyethyl)-4-(etoxycarbonyl)imidazole or anilide 1-(2-hydroxyethyl)imidazole-4-carboxylic acid

The invention relates to new N-heterocyclic derivatives of the formula (I):

where: A means-OR1-C(O)N(R1R2or-N(R1R21; each X, Y and Z independently represents N or C(R19); each U represents N or C(R5), provided that U is N only when X represents N, and Z and Y denote CR19; each W represents N or CH; V denotes: (1) N(R4); (2) C(R4)H; or (3) the groupdirectly related to the group -(C(R14R20)n-A,denotes a 5-6-membered N-heterocyclyl, optionally containing 6-membered ring additional heteroatom selected from oxygen, sulfur and NR6where R6denotes hydrogen, optionally substituted phenyl, 6-membered heterocyclyl containing 1-2 nitrogen atom, optionally substituted 5-membered heterocyclyl containing 1-2 nitrogen atom, aminosulfonyl, monoalkylammonium, dialkylaminoalkyl,1-6alkoxycarbonyl, acetyl, etc

The invention relates to organic chemistry and can find application in medicine

The invention relates to new selective antagonists1b-adrenergic receptors of the formula :

where a is a 2-tetrahydrofuryl, 2-furyl, substituted linear or branched alkyl containing from 1 to 5 carbon atoms, benzyloxy group or phenoxyalkanoic group, a substituted linear or branched alkyl containing from 1 to 5 carbon atoms and/or alkoxy group containing from 1 to 4 carbon atoms, or an enantiomer, diastereoisomer or pharmaceutically acceptable salt of such compounds, and to pharmaceutical compositions based on these compounds, which are able to improve the condition associated with sexual dysfunction

The invention relates to pharmaceutical industry and AA derivatives of N-(aryloxyalkyl)-heteroarylboronic and-heteroarylboronic General formula (I) used to obtain drugs with antipsychotic or analgesic activity, and a method of treating psychoses by using these derivatives

The invention relates to a derivative of erythromycin formulas (I)

in which Y denotes a hydrogen atom or fluorine; n denotes an integer from 1 to 8; Z represents a hydrogen atom or a residue of carboxylic acid, and in which pyrazol cycle substituted heteroaryl radical, which contains one nitrogen atom; and their salt adducts with tilotama

The invention relates to a method for producing a condensed 2-getreleasedate General formula

using the diamine of General formula

where A=

R=2-furyl, 2-thienyl, 2-(1-methyl)pyrrolyl, 3-(1-methyl)indolyl, and aldehydes in the presence of acetate or copper sulfate, characterized in that the interaction takes place by boiling in 50% acetic acid, followed by decomposition of the copper salt, the effect on its suspension in 50% acetic acid sodium thiosulfate in 100With

The invention relates to indole derivative of General formula I:

where R1represents hydrogen, halogen, methoxy; R2represents hydrogen, halogen, methyl, ethyl, methoxy; R3represents carboxy, tetrazolyl or CONHSO2R4in which R4represents methyl, ethyl, phenyl, 2,5-dimethylisoxazole, trifluoromethyl; T represents-CH2- or-SO2-; and ring a is 3-chlorophenyl, 4-chlorophenyl, 3-triptoreline, 3,4-dichlorophenyl, 3,4-differenl, 3-fluoro-4-chlorophenyl, 3-chloro-4-forfinal, 2,3-dichloride-5-yl; or their pharmaceutically acceptable salts or esters, as well as pharmaceutical compositions containing them

The invention relates to new nitrogen-containing aromatic 6-membered cyclic compounds of the formula (I) or their pharmaceutically acceptable salts, demonstrating excellent selective PDE V inhibitory activity

The invention relates to organic chemistry and agriculture, in particular to the derived SIM-triazine as a compound that protects against fetotoksicheskoe actions of herbicides in the cultivation of sunflower (antidote)

FIELD: organic chemistry, medicine, pharmacology.

SUBSTANCE: invention relates to new derivatives of carbamic acid esters of the general formula (I):

and their pharmaceutically acceptable salts eliciting activity with respect to metabotropic glutamate receptors mGlu of group I that can be used for treatment of acute and/or chronic neurological disorders. In the general formula (I) R1 means hydrogen atom or (C1-C7)-alkyl; R2 and R2' mean independently of one another hydrogen atom, (C1-C7)-alkyl, (C1-C7)-alkoxy-group, halogen atom or trifluoromethyl; X means oxygen (O), sulfur (S) atom or two hydrogen atoms not forming a bridge; A1/A2 mean independently of one another phenyl or 6-membered heterocycle comprising 1 or 2 nitrogen atom; B represents group of the formula:

wherein R3 means (C1-C7)-alkyl and others; Y means -O-, -S- or a bond; Z means -O- or -S-; or B means 5-membered heterocyclic group of formulae: (a) , (b) , (c) or (d) . Also, invention relates to methods for preparing compounds and to a medicinal agent based on thereof.

EFFECT: improved preparing methods, valuable medicinal properties of compounds.

22 cl, 1 tbl, 2 sch, 78 ex

Up!