KR20000065090A - 3-alkoxypyridine-2-carboxylic acid amide ester, preparation method thereof and its use as a pharmaceutical - Google Patents

Abstract

The present invention relates to 3-alkoxypyridine-2-carboxylic acid amide esters of formula (I) and physiologically effective salts thereof, methods for their preparation and their use for inhibiting collagen biosynthesis and their use as agents for treating fibrotic diseases. .

Formula I

In the above formula,

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is (C ₁ -C ₆ ) alkyl,

A is a -CH ₂ group wherein one hydrogen may be substituted with a methyl group,

B is -GO ₂ G, where G is a radical of GOH alcohol.

Description

3-alkoxypyridine-2-carboxylic acid amide ester, its preparation method and its use as medicament

The present invention relates to 3-alkoxypyridine-2-carboxamidoesters, methods for their preparation, and their use for inhibiting collagen biosynthesis and their use as medicaments for the treatment of fibrotic disorders.

Compounds that inhibit the enzymes prolyl and lysyl hydroxylase very selectively inhibit collagen biosynthesis by acting on collagen specific hydroxylation reactions. In this process, protein bound proline or lysine is hydroxylated by the enzyme prolyl or lysyl hydroxylase. When this reaction is inhibited by an inhibitor, a nonfunctional, insufficiently hydroxylated collagen molecule can be produced that can only be released into the extracellular space by the cell. Insufficiently hydroxylated collagen also cannot be incorporated into the collagen matrix and thus proteolyses very easily. As a result of this effect, the amount of extracellularly attached collagen is reduced overall.

Thus, inhibitors of prolyl hydroxylase are suitable materials for treating diseases in which the adhesion of collagen is critically contributing to the clinical picture. This includes especially fibrosis of the lungs, liver and skin, such as scleroderma and scars after burns, injuries and surgical operations, and also atherosclerosis.

The enzyme prolyl hydroxylase is known to be effectively inhibited by pyridine-2,4- and -2,5-dicarboxylic acids. See K. Majamaa et al., Eur. J. Biochem. 138 (1984) 239-245). However, in cell culture these compounds are active as inhibitors only at very high concentrations. Tschank, G. et al., Biochem. J. 238 (1987) 625-633.

Also known are prodrugs of pyridine-2,4 (5) -dicarboxylates. These are described in patent applications EP-A 0 590 520, DE-A 42 38 506 and EP-A 0 562 512.

N-oxalylglycine as an inhibitor of prolyl-4-hydroxylase is described by J. Med. Chem. 1992, 35, 2652-2658 (Cunliffe et al.) And EP-A 0 457 163.

In addition, 3-hydroxypyridine-2-carboxylic acid (glycosyl ethyl ester) amide is also described in J. Org. Chem. 31, 636-638 (1966).

It is therefore an object of the present invention to provide compounds which are distinguished by their high activity in particular in vivo and / or in vitro, in particular as their systemic and / or topical use.

Surprisingly, it has now been found that the 3-alcoholpyridine-2-carboxamidoester of formula I is a particularly potent inhibitor of collagen biosynthesis. The compounds of the present invention are selected from the compounds described in EP-A 0 650 960. When compared to the compounds mentioned in the European Patent Application, they are distinguished by high activity in vivo and in vitro against fibrotic diseases, in particular as systemic and / or topical. This includes, for example, fibrosis of the lungs, liver, kidneys, heart, eyes and skin, and atherosclerosis.

Compounds of formula (I) include organ culture of the most diverse cells (e.g., normal human dermal fibroblasts, primary fat storing cells between rats, epithelial cells between rats, and calvaria). Water) to potentially inhibit collagen biosynthesis.

In particular they are suitable for topical use as collagen biosynthesis inhibitors and antifibrotic active compounds, and for topical use in the form of diseases caused by increased formation of connective tissue (collagen). Thus, the compounds of formula (I) are suitable for topical use, for example for avoiding / reducing scars after surgical operations on the body, and for the postoperative treatment of ophthalmic diseases (eg glaucoma). It is suitable for topical use and in particular for topical use in radiation-induced fibrosis or chemotherapy-induced fibrosis.

The compounds according to the invention are ester prodrugs of the corresponding carboxylic acids of formula I, wherein B is a carboxyl group.

Compounds of formula (I) are degraded in living bodies (in vivo) and in cell culture (in vitro) to produce compounds of formula (I) or salts thereof, wherein B is a carboxyl group.

The compound of formula (I), after administration, forms a compound of formula (I) or a salt thereof, wherein B is a carboxyl group, thereby inhibiting collagen biosynthesis observed in vivo and in vitro. These compounds inhibit prolyl-4-hydroxylase to inhibit the biosynthesis of collagen.

The compounds according to the invention are the compounds of formula (I) and their physiologically active salts.

In the above formula,

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is (C ₁ -C ₆ ) -alkyl,

A is a -CH ₂ group wherein one hydrogen may be substituted with a methyl group,

B is —CO ₂ G, where G is a radical of alcohol GOH.

Preferred compounds of formula I and their physiologically active salts are

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is (C ₁ -C ₆ ) -alkyl,

A is a —CH ₂ group wherein one hydrogen may be substituted with a methyl group,

B is a —CO ₂ G [where G is a branched, straight or cyclic (C ₁ -C ₂₀ ) -alkyl radical or is a branched, straight or cyclic (C ₂ -C ₂₀ ) -alkenyl radical, corresponding ( A C ₂ -C ₂₀ ) -alkynyl radical, a corresponding (C ₄ -C ₂₀ ) -alkeninyl radical or a retinyl radical, where each radical comprises at least one multiple bond in each case, or phenyl Alkyl radicals, wherein the radicals are in particular hydroxyl, halogen, cyano, trifluoromethyl, carboxyl, (C ₁ -C ₁₂ ) -alkyl, (C ₃ -C ₈ ) -cycloalkyl, (C ₅ -C ₈ ) -cycloalkenyl, (C ₁ -C ₁₂ ) -alkoxy, (C ₁ -C ₁₂ ) -alkoxy- (C ₁ -C ₁₂ ) -alkyl, (C ₁ -C ₁₂ ) -alkoxy- (C ₁ -C ₁₂ ) -alkoxy, phenyl- (C ₁ -C ₄ ) -alkyloxy, (C ₁ -C ₈ ) -hydroxyalkyl, (C ₁ -C ₁₂ ) -alkylcarbonyloxy, (C ₃ -C ₈ ) -cycloalkylcarbonyloxy, benzoyloxy and phenyl- (C ₁ -C ₄ ) -alkylcarbonyloxy One or more substituents), and physiologically active salts thereof.

Further preferred compounds of formula I and their physiologically active salts are

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is (C ₁ -C ₄ ) -alkyl,

A is a —CH ₂ group wherein one hydrogen atom may be substituted with a methyl group,

B is —CO ₂ G, wherein G is a branched, straight or cycloaliphatic (C ₁ -C ₁₈ ) -alkyl radical, (C ₃ -C ₈ ) -cycloalkyl- (C ₁ -C ₈ ) -alkyl radical or , Branched or straight chain (C ₂ -C ₁₈ ) -alkenyl radicals (eg geranyl, farnesyl or retinyl radicals) or the corresponding (C ₂ -C ₁₈ ) -alkynyl radicals, benzyl, phenethyl, Phenylpropyl or phenylbutyl radicals wherein the radicals are hydroxyl, (C ₁ -C ₄ ) -alkoxy, (C ₁ -C ₆ ) -alkylcarbonyloxy, (C ₃ -C ₈ ) -cycloalkylcarbonyloxy , Benzoyloxy or phenyl- (C ₁ -C ₄ ) -alkylcarbonyloxy); and a physiologically active salt thereof.

Particularly preferred compounds of formula I and their physiologically active salts are

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is (C ₁ -C ₄ ) -alkyl,

A is a —CH ₂ group wherein one hydrogen atom may be substituted with a methyl group,

B is a —CO ₂ G wherein G is a branched, straight or alicyclic (C ₁ -C ₁₈ ) -alkyl radical, (C ₃ -C ₈ ) -cycloalkyl- (C ₁ -C ₄ ) -alkyl radical, Branched or straight chain (C ₂ -C ₁₈ ) -alkenyl radicals, benzyl, phenethyl, phenylpropyl or phenylbutyl radicals) and physiologically active salts thereof.

Furthermore, particularly preferred compounds of formula I and their physiologically active salts are

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is methyl,

A is a —CH ₂ group wherein one hydrogen atom may be substituted with a methyl group,

B is a -CO ₂ G wherein G is a branched or straight chain (C ₁ -C ₁₈ ) -alkyl or (C ₂ -C ₁₈ ) -alkenyl radical) and a physiologically active salt thereof.

Furthermore, particularly preferred compounds of formula I and their physiologically active salts are

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is methyl,

A is a -CH ₂ group,

B is a -CO ₂ G wherein G is a branched or straight chain (C ₁ -C ₁₈ ) -alkyl or (C ₂ -C ₁₈ ) -alkenyl radical) and a physiologically active salt thereof.

Particularly preferred compounds of formula I and their physiologically active salts are

R ¹ , R ² and R ³ are hydrogen,

R ⁴ is methyl,

A is a -CH ₂ group,

B is a -CO ₂ G wherein G is a linear (C ₁ -C ₁₈ ) -alkyl radical and a physiologically active salt thereof.

Particularly preferred compounds of formula I and their physiologically active salts are compounds wherein G is a linear (C ₁ -C ₁₆ ) -alkyl radical and physiologically active salts thereof.

The present invention also includes salts of compounds of formula (I).

Forming salts with acidic reagents can be carried out at the N atom of pyridine.

Reagents used are, for example, drugs containing toluenesulfonic acid, methanesulfonic acid, HCl, H ₂ SO ₄ , H ₃ PO ₄ and acidic groups.

The present invention relates to compounds of formula (I) and physiologically acceptable salts thereof for use in inhibiting collagen biosynthesis, in particular in collagen biosynthesis in different cells.

The present invention relates to compounds of formula (I) and physiologically acceptable salts thereof for use in inhibiting prolyl-4-hydroxylase in vivo and in vitro.

The present invention also relates to compounds of formula (I) and physiologically acceptable salts thereof for use in fibrotic diseases of the lungs, liver, kidneys, heart, eyes and skin. The compounds can also be used for atherosclerosis. In this regard, systemic and / or topical applications are used.

In particular, the present invention is intended for topical use, in particular for use as a collagen biosynthesis inhibitor, an antifibrotic active compound, and for topical use in a form of disease caused by increased formation of connective tissue (collagen). It relates to compounds and physiologically acceptable salts thereof. It is used for avoiding / reducing scars after surgical surgery on the body, and for postoperative treatment of eye diseases (e.g. glaucoma) and for fibrosis caused by fibrosis or chemotherapy caused by radiation in the lung in particular. Use is included.

Finally, the present invention relates to a compound of formula (I) for use as a medicament.

In particular, the present invention relates to a compound of formula (I) for topical use in fibrosis of the skin, lungs and eyes, and in particular for topical treatment after surgery in glaucoma.

The present invention also relates to a process for the preparation of the compound of formula (I).

A compound of Formula I wherein A is a -CH ₂ -group wherein one hydrogen atom may be substituted with a methyl group and B is CO ₂ G

Iii) pyridine-2-carboxylic acid of Compound II (R ⁵ = H) with an amino ester of Compound III or a salt thereof to give an amidoester of Formula I, or

Ⅰ2) compound _{^{Ⅱ (R 5 = (C 1}} -C 16 - alkyl) by reacting the 2-carboxylic acid ester of a) under aminolysis conditions or to obtain a compound of formula Ⅰ,

Ii) esterification of compound IV with alcohol GOH, or

Iii) Compound V is prepared by alkylation with R ⁴ X, wherein X is a leaving group, in particular halogen, -OSO ₂ Me, -OSO ₂ phenyl and the like.

Suitable methods for the amide formation (reaction # 1) are the carbonyl activation and condensation reaction methods known in peptide chemistry.

Reagents used for carboxylic acid activation may be substances known to those skilled in the art, for example thionyl chloride, oxalyl chloride, pivaloyl chloride, chloroformic acid ester derivative or N, N'-carbonyldiimidazole. The activated derivative of compound II is prepared and reacts in situ with the amide derivative of compound III.

Suitable condensing agents are, for example, mixtures of N, N'-dicyclohexylcarbodiimide / N-hydroxy-1H-benzotriazole and N-ethylmorpholine.

Suitable solvents are dichloromethane, tetrachloromethane, butyl acetate, ethyl acetate, toluene, tetrahydrofuran, dimethoxyethane, 1,4-dioxane, acetonitrile, N, N-dimethylformamide, N, N-dimethylacet Amide, dimethyl sulfoxide, nitromethane and / or pyridine.

The compounds of formula (I) according to the invention have valuable pharmacological properties and in particular exhibit antifibrotic activity.

Antifibrotic activity can be measured in the following animal models:

Connective tissue capsule formation around an osmotic minipump implanted subcutaneously [Unemori, E.N. et al., J. Invest. Dermatol., 101: 280-5, 1993;

Collagen content of "cotton pellets" or polyvinyl sponges implanted subcutaneously (Boyle, E., Mangan, F.R., Br. J. Ep. Pathnol. 61: 351-60, 1980; and

Collagen content of the lungs after radiation or chemically induced fibrosis [Ward, H.E et al., Res., 136, 15-21, 1993 and also Santana, A. et al., Am. J. Respir. Cell. Mol. Biol. 13: 34-44, 1995].

Inhibition of prolyl-4-hydroxylase in cell culture:

Normal human dermal fibroblasts (NHDF), rat liver epidermal cells (Ref. 1) and hepatic fat storage cells (Ref. 2) are used as cell models for material testing. To do this, the cells are cultured in the presence of inhibitors. At the same time, the newly synthesized collagen is metabolically labeled by 4- ³ HL-proline and ¹⁴ C-proline. The effect of the test substance on the degree of hydroxylation of collagen is then measured according to the method of Kojkier et al. (Ref. 3). The reference material used is 2,2'-dipyridyl [1: Schrode, W., Mecke, D., Gebhard, R. 1990. Induction of glutamine synthetase in periportal hepatocytes by co-cultivation with a liver epithelial cell line. Eur. J. Cell. Biol. 53: 35-41, 2 .: Blomhoff, R., Berg T. 1990. Isolation and cultivation of rat liver stellate cells. Methods Enzymol. 190: 59-71 and 3 .: Chojkier, M. Peterkofsky, B., Bateman, J. 1980. A new method for determining the extent of proline hydroxylation by measuring changes in the ration of [4- ³ H]: [ ¹⁴ C] proline in collagenase digests. Anal. Biochem. 108: 385-393.

Inhibition of prolyl-4-hydroxylase in vitro (chicken fetal cranial tube) according to literature [Biochem. J. (1994) 300 525-300]:

1. Metabolic Labeling

Cut the cranial canal of a 15-day-old chicken fetus and for 3 minutes at 37 ° C. in Hanks' balanced salt solution minimum essential medium Eagle, HMEM, BioWhittaker, Walkersville, MD, USA. Wash. Four cranial tubes each were incubated for 2.5 hours at 37 ° C. in a glass tube with 2 mM glutamine, 1 μCi of [U- ¹⁴ C] proline and 1.5 ml of HMEM added with various concentrations of inhibitor. The incubation is terminated by inserting the sample tube into the ice. The medium is removed and the cranial tube is briefly washed with 1 ml of secondary distilled H ₂ O. The cranial tubes were treated with 3 ml of 0.5 M acetic acid and 18 μg of phenylmethanesulfonyl fluoride and then extracted for 16 hours. The extraction solution is dialyzed at 4 ° C. against 0.5 M acetic acid to remove free [U- ¹⁴ C] proline, and then freeze dried in aliquots.

2. Hydroxyproline-Analysis

An aliquot of the lyophilized sample is dissolved in 2 ml of 6N HCl and hydrolyzed at 105 ° C. for 24 hours. Hydrochloric acid is then evaporated. The residue is resuspended in 300 μl double distilled H ₂ O, transferred to an Eppendorf tube and dried again. The remaining hydrochloric acid is dissolved in 60 µl of ethanol / H ₂ O / triethylamine 2: 2: 1 (v: v: v :), neutralized and dried repeatedly. The amino acid is then derived for 20 minutes at room temperature in a 7: 1: 1: 1 (v: v: v: v) solution of ethanol / triethylamine / phenyl isothiocyanate / H ₂ O and dried again. For analysis by HPLC, the samples are dissolved in 150 μl of phosphate buffer (5 mM Na ₂ HPO ₄ (pH 7.4) / acetonitrile 95: 5 (v: v)) and centrifuged at 10,000 g for 5 minutes. 50 μl is used for the analysis. HPLC chromatography is performed on a C18 reverse phase column ultrasphere ODS (Ultrasphere ODS, 4.6 min × 7.5 cm, Beckman) 3 μm at 50 ° C. using the following gradient system.

Time Buffer A Buffer B

0 minutes 100%

0 to 9 minutes 100 to 90% 0 to 10%

9 to 11 minutes 90 to 0% 10 to 100%

11 to 12 minutes 0% 100%

12 to 14 minutes 0 to 100% 100 to 0%

14-19 minutes 100%

Solution A: 70 mM sodium acetate (pH = 6.14) / 0.1% acetonitrile

Solution B: acetonitrile / methanol / water 45:15:40 (v: v: v)

3. SDS polyacrylamide-gel electrophoresis

Lyophilized sample aliquots from item 1 above were dissolved using 10 mM Tris / HCl (pH 8) / 1 mM EDTA / 1% SDS and 5% mercaptoethanol for SDS polyacrylamide-gel electrophoresis, 95 Denature at 5 ° C. Electrophoresis is performed on linear gradient gels (5-15%). After fixing in 3: 1: 6 (v: v: v) in methanol / glacial acetic acid / water, the gel is dried and exposed to Kodak X-Omat film at −70 ° C.

The following is a comparison by inhibition of prolyl-4-hydroxylase (collagen biosynthesis) in liver cells of rats that the compounds of the present invention have surprising advantages when compared to the compounds of EP-A 0 650 960. Experiment proved.

Example Compound Inhibition rate of collagen biosynthesis in rat liver cells Compounds of the Examples of EP-A 0 650 960 Fat storage cells between rats 27151154155239 -79%: 100㎛-65%: 25㎛-40%: 25㎛-59%: 50㎛-41%: 25㎛ Compounds of the Examples of EP-A 0 650 960 Epidermal Cells Between Rats 290288 -86%: 25㎛ -72%: 25㎛

Example Compound Inhibition rate of collagen biosynthesis in rat liver cells Compounds of the Examples of the Invention Fat storage cells between rats 2 -50%: 1.5 + 0.1㎛ 3 -50%: 0.9 + 0.1㎛ 14 -50%: 10.0㎛ Compounds of the Examples of the Invention Epidermal Cells Between Rats 214 -50%: 7㎛-50%: 10.0㎛

Compounds of the Examples of the Invention Cranial tube 2 -50%: 0.032㎛ 14 -50%: 0.41㎛ Normal human fibroblasts (NHDF) 123 -50%: 0.1㎛-50%: 0.2㎛-50%: 0.5㎛

The results of the comparison show that the compounds according to the invention of the present invention show high inhibition (-50%) even at surprisingly low concentrations (depending on the test model, 10 μm or less) when compared to the compounds of EP-A 0 650 960 present.

The compounds of formula (I) may be used as medicaments in the form of pharmaceutical preparations which optionally contain an acceptable pharmaceutical vehicle. The compounds can be used as therapeutic agents, for example, pharmaceutical organic or inorganic excipients suitable for enteric, transdermal or parenteral administration or for inhalation administration (e.g. water, gum arabic, gelatin, lactose, starch, magnesium stearate, Talc, vegetable oils, polyalkylene glycols, petroleum jelly, and the like), and as pharmaceutical preparations containing these compounds.

For this purpose, they are orally 0.1 to 25 mg / kg per day, preferably 1 to 5 mg / kg or parenterally 0.01 to 5 mg / kg per day, preferably 0.01 to 2.5 mg per day. / Kg, in particular 0.5 to 1.0mg / kg per day. In severe cases, the dose may also be increased. In many cases, however, lower doses are sufficient. The above description relates to an adult weighing about 75 kg.

3-methoxypyridine-2-carboxylic acid (Compound II; R ¹ to R ³ = H, R ⁴ = Me, R ⁵ = H) is known in the literature. EV Brown, MB Shambhu, J. Org . Chem. 36, 2002 (1971).

In addition, methyl 3-methoxypyridine-2-carboxylate (Compound II; R ⁵ = CH ₃ ) is known in the literature. Acta Chem. Scand. 23, 1791 (1969).

3-methoxypyridine-2-carboxylic acid used as starting material in the following examples is obtained starting from 4-chloro-3-methoxy-2-methylpyridine obtained from maltol. A 0 208 452 and EP-A 0 304 732].

In the examples to be described below, the compounds of formula (I) according to the invention are referred to as substituted pyridine-2-carboxylic acid (glycyl ester) amides.

This notation method is understood to mean substituted pyridine-2-carboxylic acid N-((alkoxycarbonyl) methyl) amide.

It is also possible to classify into substituted N- (pyridyl-2-carbonyl) glycines.

Example 1

N-(((1-butyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

a) 3-methoxypyridine-2-carboxylic acid hydrochloride

a.1) 4-Chloro-3-methoxy-2-methylpyridine 1-oxide

11.2 g (80.5 mmol) of 3-methoxy-2-methyl-4 (1H) -pyridone in 100 mL of phosphorus oxychloride is heated under reflux for 10 hours.

The mixture is subsequently concentrated, 2 ml each is mixed with 30 ml of toluene and concentrated again, the residue is dissolved in water and the pH is adjusted to 11 with K ₂ CO ₃ and extracted with dichloromethane and the organic phase is extracted. Wash with water and dry to evaporate the solvent. The resulting pale brown oil (9 g) is used to obtain 8 g of product (m .: 88-89 ° C., from petroleum ether) using m-chloroperbenzoic acid in dichloromethane under standard conditions.

a.2) 4-chloro-2-hydroxymethyl-3-methoxypyridine

30 g (173 mmol) of 4-chloro-3-methoxy-2-methylpyridine N-oxide are dissolved in 100 ml of ice-cold acetic acid, mixed with 150 ml of acetic anhydride dropwise with stirring at 80 캜, and the mixture is mixed at 110 캜. Stir for 2 hours. After the mixture was cooled to 80 ° C., 200 ml of methanol was added dropwise, the mixture was heated for 15 minutes to boil and cooled, then concentrated under reduced pressure, the residue was dissolved in methanol and dissolved in 300 ml of 1.5N methanolic sodium hydroxide solution. Pour, stir at 20 ° C. for 30 min and concentrate under reduced pressure, dissolve the residue in water and extract three times with dichloromethane, dry and concentrate the organic phase and crystallize the residue with petroleum ether. This gives 23 g of product (melting point: 64 to 66 ° C.).

a.3) 4-chloro-3-methoxypyridine-2-carboxylic acid

At 20 ° C., 52.2 g (0.3 mol) of the alcohol is added to 24 g of KOH dissolved in 900 ml of water, and the mixture is briefly mixed with 70 g (0.44 mol) of potassium permanganate, respectively, at once at 50 to 60 ° C. until discolored. After stirring for 1 h at 50 ° C., the hot mixture is filtered off with suction, the filter cake is washed three times with hot water, concentrated to 300 ml under reduced pressure, and the pH is adjusted to 1 with concentrated HCl while cooling. . It is suction filtered and dried to give 50 g of product (melting point: 121 ° C., decomposed).

a.4) 29 g (0.15 mol) of the 4-chloropyridinecarboxylic acid in 500 ml of methanol / tetrahydrofuran (1: 1) are hydrogenated to Pd / C (10%) in a hydrogenation apparatus. 3.1 L of hydrogen is absorbed, the catalyst is removed by suction filtration, the filtrate is concentrated, the crystalline residue is treated with ethyl acetate, suction filtered and dried to give 29 g of product (melting point: 170 ° C., decomposed).

b) 3.8 g (20 mmol) of 3-methoxypyridine-2-carboxylic acid hydrochloride are suspended in 500 mL of anhydrous dichloromethane and glycine 1-butyl ester tosylate (toluene with water to glycine with stirring at 20 ° C.) , Prepared from 1-butanol and p-toluenesulfonic acid) 6.1 g (20 mmol), followed by 7.5 ml (60 mmol) of N-ethylmorpholine, 3 g (22.5 m) 1-hydroxy-1H-benzotriazole. Mol) and N-cyclohexyl-N '-(2-morpholinoethyl) carbodiimide meto-p-toluenesulfonate (CMC) 8.5 g (20 mmol) are mixed and stirred at 20 ° C for 24 hours. . Undissolved particles are suction filtered and the filtrate is extracted successively with aqueous sodium bicarbonate solution, 1N aqueous hydrochloric acid and water. The organic phase is dried and concentrated under reduced pressure, and the oily residue is scraped off to crystallize. This affords 1.76 g of a colorless title compound (melting point: 60-62 ° C.).

Example 2

N-(((1-octyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

2.5 g (13 mmol) of 3-methoxypyridine-2-carboxylic acid hydrochloride, 5.5 ml (45 mmol) of N-ethylmorpholine, 2 g (15 mmol) of 1-hydroxy-1H-benzotriazole, glycine octyl ester tosyl 4.7 g (13 mmol) of rate (prepared on a water separator using glycine, 1-octanol and p-toluenesulfonic acid) and 6.3 g (15 mmol) of CMC (see Example 1) in 350 ml of anhydrous dichloromethane Stir for 48 hours. After working up by the method of Example 1, the crude product is chromatographed on silica gel with dichloromethane (up to 2.5% methanol is added during elution). This gives 3.6 g of a colorless oily title compound. ¹ H NMR (CDCl ₃ ): δ = 4.26 (d, CH ₂ -glycine).

Example 3

N-(((1-1-decyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

By the method of Example 1, 2.8 g (15 mmol) of 3-methoxypyridine-2-carboxylic acid hydrochloride in 400 ml of anhydrous dichloromethane was converted to glycine 1-dodecyl ester tosylate (38 g (0.5 mol) of glycine, 1-). Prepared from 93.2 g (0.5 mol) of dodecanol and 115 g (0.6 mmol) of p-toluenesulfonic acid, 140 g of the product is crystallized from diethyl ether and sintered at 80 ° C. Melting point: about 106 ° C. 6.2 g (15 mmol) ), 8.2 g (60 mmol) N-ethylmorpholine, 2 g (15 mmol) 1-hydroxy-1H-benzotriazole and 6.3 g (15 mmol) CMC are stirred at 20 ° C. for 24 hours. Since it forms a strong emulsion, dichloromethane is distilled off under reduced pressure and the residue is dissolved in diethyl ether for post-treatment similar to Example 1. The organic phase is dried over magnesium sulfate and concentrated under reduced pressure, and then 4.7 g of the resulting yellow oil is chromatographed on silica gel using ethyl acetate. This gives 4 g of pale yellow oil, which is scraped off with a glass rod and left to stand for 2 days before crystallization. 3.6 g of the colorless title compound are isolated (melting point: 47-49 ° C.).

Example 4

N-(((1-pentyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 5

N-(((1-hexyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 6

N-(((1-heptyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 7

N-(((1-nonyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 8

N-(((1-decyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 9

N-(((1-tetradecyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 10

N-(((1-octadecyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 11

N-(((1-propyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 12

N-(((2-propyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 13

N-(((1-tridecyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 14

N-(((1-1-hexadecyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

5.7 g (30 mmol) of 4-chloro-3-methoxypyridine-2-carboxylic acid and glycine hexadecyl ester tosylate (melting point; approx. 90 ° C., glycine, 1-hexa on a water separator using toluene in 300 ml of dichloromethane) 14.2 g (30 mmol) of N-ethylmorpholine, 4.5 g of 1-hydroxy-1H-benzotriazole, was prepared according to the method of Example 1. (33 mmol) and N-cyclohexyl-N '-(2-morpholinoethyl) carbodiimide meto-p-toluenesulfonate (CMC) are mixed together with 12.8 g (30 mmol) and stirred for 24 hours. . The undissolved particles are suction filtered, the filtrate is extracted with aqueous sodium bicarbonate solution, water and aqueous hydrochloric acid, the organic phase is concentrated, the residue (14 g) is dissolved in tetrahydrofuran / methanol (1: 1) and Pd / Mix with C (10%) and hydrogenate in hydrogenation apparatus.

After terminating the hydrogen uptake, the catalyst is suction filtered, the filtrate is concentrated and the residue is chromatographed on silica gel with ethyl acetate. The appropriate fractions are concentrated and the residue is crystallized using diisopropyl ether. This affords 2.1 g of the title compound as a colorless substance (melting point: 63 to 65 ° C).

Example 15

N-((ethyloxycarbonyl) methyl) -3-methoxypyridine-2-carboxamide hydrochloride

a) N-((ethyloxycarbonyl) methyl) -4-chloro-3-methoxypyridine-2-carboxamide

For preparation, 4.7 g (25 mmol) of 4-chloro-3-methoxypyridine-2-carboxylic acid are suspended in 200 ml of dichloromethane anhydride, 3.5 g (25 mmol) of glycine ethyl ester hydrochloride with stirring at 20 ° C., Continuous with 6.4 mL (50 mmol) of N-ethylmorpholine, 3.8 g (28 mmol) of 1-hydroxy- (1H) -benzotriazole and 5.15 g (25 mmol) of N, N'-dicyclohexylcarbodiimide Mixed and stirred at 20 ° C. for 20 hours. The undissolved particles are filtered and the organic phase is shaken with saturated aqueous sodium bicarbonate solution, dried under reduced pressure and concentrated, and the residue (6 g of oil) is chromatographed on silica gel with ethyl acetate to give 5.4 g of an oily product. .

b) The title compound is obtained by hydrogenation of the material to Pd / C (10%) in a hydrogenation apparatus (melting point: 141 to 142 ° C., gas evolution, from diethyl ether)

Example 16

N-(((2-nonyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide racemate

2.5 g (10 mmol) of N- (carboxymethyl) -3-methoxypyridine-2-carboxamide hydrochloride (melting point: 157 DEG C, gas evolution) were suspended in 100 ml of anhydrous tetrahydrofuran, and triethylamine 1.6 After mixing with 12 ml (12 mmol), 2.4 g of pivaloyl chloride dissolved in a small amount of tetrahydrofuran is added dropwise while stirring (temperature rises to 35 to 40 ° C). After 30 minutes, the mixture was concentrated under reduced pressure, the red residue was dissolved in 100 ml of anhydrous tetrahydrofuran, mixed with 1.6 ml of triethylamine, and the mixture was stirred at 20 ° C. in sodium 2-nonanol in 2-nonanol. To a solution of latex (prepared from 30 mL 2-nonanol and 0.8 g (20 mmol) NaH). After 1 h, the mixture was concentrated under reduced pressure, the residue was mixed with dichloromethane, extracted with 2N aqueous ammonium chloride solution, the organic phase was dried under reduced pressure and concentrated, and the residue (9 g) was purified on silica gel with ethyl acetate. Chromatography. This gives 1.1 g of a colorless oily title compound. 'H NMR (DMSO): δ = 3.95 (d, CH ₂ -glycine).

Example 17

N-(((4-heptyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

2.5 g (10 mmol) of N- (carboxymethyl) -3-methoxypyridine-2-carboxamide hydrochloride were treated as in Example 16 and then sodium 4-heptanol in 4-heptanol at 20 ° C. Mix with 140 ml of a solution of Rate (prepared from 140 ml 4-heptanol and 0.6 g (25 mmol) sodium in an ultrasonic bath). After 30 minutes, the mixture is heated at 70-80 ° C. for 1 hour, cooled and concentrated under reduced pressure, the residue is dissolved in water and extracted with dichloromethane, the organic phase is concentrated under reduced pressure and dried in an oil pump. After about 15 hours the oily crude product crystallizes (melting point: 75-78 ° C.).

Example 18

N-(((3-pentyloxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 19

N-(((1- (cis-9-octadecenyl) oxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 20

N-(((1- (trans-3-hexenyl) oxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 21

N-(((1- (3-methylbutyl) oxy) carbonyl) methyl) -3-methoxypyridine-2-carboxamide

Example 22

N-((methyloxycarbonyl) methyl) -3-methoxypyridine-2-carboxamide

Claims (22)

Compounds of Formula I and their physiologically active salts. Formula I In the above formula, R ¹ , R ² and R ³ are hydrogen, R ⁴ is (C ₁ -C ₆ ) -alkyl, A is a -CH ₂ group wherein one hydrogen may be substituted with a methyl group, B is —CO ₂ G, where G is a radical of alcohol GOH. The method of claim 1, R ¹ , R ² and R ³ are hydrogen, R ⁴ is (C ₁ -C ₆ ) -alkyl, A is a —CH ₂ group wherein one hydrogen may be substituted with a methyl group, B is a —CO ₂ G [where G is a branched, straight or cyclic (C ₁ -C ₂₀ ) -alkyl radical or is a branched, straight or cyclic (C ₂ -C ₂₀ ) -alkenyl radical, corresponding ( A C ₂ -C ₂₀ ) -alkynyl radical, a corresponding (C ₄ -C ₂₀ ) -alkeninyl radical or a retinyl radical, where each radical comprises at least one multiple bond in each case, or phenyl Alkyl radicals, wherein the radicals are in particular hydroxyl, halogen, cyano, trifluoromethyl, carboxyl, (C ₁ -C ₁₂ ) -alkyl, (C ₃ -C ₈ ) -cycloalkyl, (C ₅ -C ₈ ) -cycloalkenyl, (C ₁ -C ₁₂ ) -alkoxy, (C ₁ -C ₁₂ ) -alkoxy- (C ₁ -C ₁₂ ) -alkyl, (C ₁ -C ₁₂ ) -alkoxy- (C ₁ -C ₁₂ ) -alkoxy, phenyl- (C ₁ -C ₄ ) -alkyloxy, (C ₁ -C ₈ ) -hydroxyalkyl, (C ₁ -C ₁₂ ) -alkylcarbonyloxy, (C ₃ -C ₈ ) -cycloalkylcarbonyloxy, benzoyloxy and phenyl- (C ₁ -C ₄ ) -alkylcarbonyloxy At least one substituent), and a physiologically active salt thereof. The method according to claim 1 or 2, R ¹ , R ² and R ³ are hydrogen, R ⁴ is (C ₁ -C ₄ ) -alkyl, A is a —CH ₂ group wherein one hydrogen atom may be substituted with a methyl group, B is —CO ₂ G, wherein G is a branched, straight or cycloaliphatic (C ₁ -C ₁₈ ) -alkyl radical, (C ₃ -C ₈ ) -cycloalkyl- (C ₁ -C ₈ ) -alkyl radical or , Branched or straight chain (C ₂ -C ₁₈ ) -alkenyl radicals (eg geranyl, farnesyl or retinyl radicals) or the corresponding (C ₂ -C ₁₈ ) -alkynyl radicals, benzyl, phenethyl, Phenylpropyl or phenylbutyl radicals, where these radicals are hydroxyl, (C ₁ -C ₄ ) -alkoxy, (C ₁ -C ₆ ) -alkylcarbonyloxy, (C ₃ -C ₈ ) -cycloalkylcarbonyl Oxy, benzoyloxy and phenyl- (C ₁ -C ₄ ) -alkylcarbonyloxy); and a physiologically active salt thereof. The method according to any one of claims 1 to 3, R ¹ , R ² and R ³ are hydrogen, R ⁴ is (C ₁ -C ₄ ) -alkyl, A is a —CH ₂ group wherein one hydrogen atom may be substituted with a methyl group, B is a —CO ₂ G wherein G is a branched, straight or alicyclic (C ₁ -C ₁₈ ) -alkyl radical, (C ₃ -C ₈ ) -cycloalkyl- (C ₁ -C ₄ ) -alkyl radical, A branched or straight chain (C ₂ -C ₁₈ ) -alkenyl radical, benzyl, phenethyl, phenylpropyl or phenylbutyl radical) and a physiologically active salt thereof. The method according to any one of claims 1 to 4, R ¹ , R ² and R ³ are hydrogen, R ⁴ is methyl, A is a —CH ₂ group wherein one hydrogen may be substituted with a methyl group, A compound of formula I and a physiologically active salt thereof, wherein B is —CO ₂ G, wherein G is a branched or straight chain (C ₁ -C ₁₈ ) -alkyl or (C ₂ -C ₁₈ ) -alkenyl radical). The method according to any one of claims 1 to 5, R ¹ , R ² and R ³ are hydrogen, R ⁴ is methyl, A is a -CH ₂ group, A compound of formula I and a physiologically active salt thereof, wherein B is —CO ₂ G, wherein G is a branched or straight chain (C ₁ -C ₁₈ ) -alkyl or (C ₂ -C ₁₈ ) -alkenyl radical). The method according to any one of claims 1 to 6, R ¹ , R ² and R ³ are hydrogen, R ⁴ is methyl, A is a -CH ₂ group, A compound of formula I and a physiologically active salt thereof, wherein B is —CO ₂ G, wherein G is a linear (C ₁ -C ₁₈ ) -alkyl radical. Reacting a pyridine-2-carboxylic acid of compound II (R ⁵ = H) with an amino ester of compound III or a salt thereof to obtain an amidoester of formula (I), or A pyridine-2-carboxylic acid ester of compound II (R ⁵ = (C ₁ -C ₁₆ -alkyl)) is reacted under aminodegradation conditions to give a compound of formula I (X2) or Esterifying compound IV with alcohol GOH (ii) or Wherein the compound V comprises alkylation with R ⁴ X, wherein X is a leaving group, in particular halogen, -OSO ₂ Me, -OSO ₂ phenyl and the like (iii), wherein A is a -CH ₂ group , Hydrogen may be substituted with a methyl group) and B is CO ₂ G. A process for the preparation of a compound of formula (I) as claimed in claim 1. In the above scheme, R ¹ , R ² , R ³ , R ⁴ , A and B are as defined in claim 1. 8. Compounds of formula I according to any one of claims 1 to 7, for inhibiting prolyl-4-hydroxylase in vivo. 8. Compounds of formula I according to any one of claims 1 to 7, for inhibiting collagen biosynthesis. 8. Compounds of formula I according to any one of claims 1 to 7, for avoiding / reducing adhesion of connective tissue (collagen) in organs (fibrosis) of the diseased human. 8. Compounds of formula I according to any one of claims 1 to 7, for use in fibrotic diseases. 13. The compound of formula I according to claim 12 for use in fibrotic diseases of the lungs, liver, kidneys, heart, eyes and skin and atherosclerosis. The compound of formula (I) according to claim 12 or 13 for use locally and / or systemically. The compound of formula (I) according to claim 14 for topical use in avoiding / reducing scars after surgical operations on the body. The compound of formula I according to any one of claims 9 to 14 for topical use in cases where the formation of connective tissue in the skin is increased (fibrosis). The compound of formula I according to any one of claims 9 to 14 for topical use (by inhalation) when the formation of connective tissue in the lungs is increased (fibrosis). The compound of formula I according to any one of claims 9 to 14 for topical use in cases where the formation of connective tissue in the eye is increased (fibrosis). The compound of formula (I) according to any one of claims 9 to 14 for topical use in the eye for the treatment after surgery of glaucoma. The compound of formula (I) according to any one of claims 9 to 14, for topical use in radiation induced fibrosis or chemotherapy induced fibrosis, in particular pulmonary fibrosis. 9. A medicament comprising at least one compound of formula (I) as claimed in any one of claims 1-8. Use of a compound for inhibiting prolyl-4-hydroxylase in fibrotic disease.