CA2253282A1 - 3-alkoxypyridine-2-carboxylic acid amide esters, their preparation and their use as drugs - Google Patents
3-alkoxypyridine-2-carboxylic acid amide esters, their preparation and their use as drugs Download PDFInfo
- Publication number
- CA2253282A1 CA2253282A1 CA002253282A CA2253282A CA2253282A1 CA 2253282 A1 CA2253282 A1 CA 2253282A1 CA 002253282 A CA002253282 A CA 002253282A CA 2253282 A CA2253282 A CA 2253282A CA 2253282 A1 CA2253282 A1 CA 2253282A1
- Authority
- CA
- Canada
- Prior art keywords
- formula
- compound
- radical
- group
- alkyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom 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
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/81—Amides; Imides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
Landscapes
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Heart & Thoracic Surgery (AREA)
- Ophthalmology & Optometry (AREA)
- Gastroenterology & Hepatology (AREA)
- Pulmonology (AREA)
- Cardiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Pyridine Compounds (AREA)
Abstract
Described are 3-alkoxypyridine-2-carboxylic acid amide esters of formula (I), wherein R1, R2 and R3 are hydrogen, R4 is a (C1-C6) alkyl, A is a -CH2 group in which one hydrogen can be replaced by a methyl group, and B is -CO2G, where G is the radical of a GOH alcohol, including the physiologically effective salts; their preparation, their use in inhibiting collagen biosynthesis, and their use as drugs for treating fibrotic diseases.
Description
CA 022~3282 1998-10-29 3-Alkoxypyridine-2-carboxamidoesters, their preparation and their use as pharmaceuticals The invention relates to 3-alkoxypyridine-2-carboxamidoesters, their 5 preparation and their use for the inhibition of collagen biosynthesis and their use as pharmaceuticals for the treatment of fibrotic disorders.
Compounds which inhibit the enzymes prolyl and Iysyl hydroxylase cause a very selective inhibition of collagen biosynthesis by affecting the collagen-10 specific hydroxylation reactions. In the course thereof, protein-bound proline or Iysine is hydroxylated by the enzymes prolyl or Iysyl hydroxylase.
If this reaction is suppressed by inhibitors, a nonfunctional, underhydroxylated collagen molecule results, which can be released into the extracellular space only in a small amount by the cells. The 15 underhydroxylated collagen additionally cannot be incorporated into the collagen matrix and is very easily proteolytically degraded. As a result of these effects, the amount of extracellularly deposited collagen is decreased overall.
20 Inhibitors of prolyl hydroxylase are therefore suitable substances in the therapy of disorders in which the deposition of collagens decisively contributes to the clinical picture. These include, inter alia, fibroses of the lung, liver and skin (scleroderma and scars after burns, injuries and surgical interventions) and also atherosclerosis.
It is known that the enzyme prolyl hydroxylase is effectively inhibited by pyridine-2,4- and -2,5-dicarboxylic acid (K. Majamaa et al., Eur. J.
Biochem.138 (1984) 239-245). In cell culture, however, these compounds are only active as inhibitors in very high concentrations (Tschank, G. et al., 30 Biochem. J. 238 (1987) 625-633).
Prodrugs of the pyridine-2,4(5)-dicarboxylates are also known. These are described in the Patent Applications EP-A-0 590 520 and DE-A42 38 506 and EP-A-0 562 512.
35 N-Oxalylglycines as inhibitors of prolyl-4-hydroxylase are known from J.Med.Chem.1992, 35, 2652-2658 (Cunliffe et al.) and EP-A-0 457 163.
~ . . . .
CA 022~3282 1998-10-29 Furthermore, 3-hydroxypyridine-2-carboxylic acid (glycyl ethyl ester)amide is described in J. Org. Chem. 31, 636-638 (1966).
It was therefore the object to provide compounds which are distinguished 5 by a particularly high in vivo and/or in vitro activity, in particular in their systemic and/or local use.
Surprisingly, it has now been found that 3-alkoxypyridine-2-carboxamidoesters of the formula I are particularly strongly active inhibitors 10 of collagen biosynthesis. The compounds of the invention are selected from the compounds described in EP-A-0 650 960. Compared with the compounds mentioned in the European Patent Application, they are distinguished by a particularly high in vivo and in vitro activity, in particular in their systemic and/or local use against fibrotic disorders. These include, 15 for example, fibroses of the lung, the liver, the kidney, the heart, the eye and the skin and in atherosclerosis.
The compounds of the formula l lead to a potent inhibition of collagen biosynthesis in the most diverse cells (e.g. normal human dermal 20 fibroblasts, primary fat storing cells from rat liver, rat liver epithelial cells and in organ cultures of calvaria).
They are particularly suitable for local use as inhibitors of collagen biosynthesis, for local use as antifibrotic active compounds and for local 25 use in forms of disease which are caused by an increased formation of connective tissue (collagen). Thus the compounds of the formula I are suitable, for example, for local use for the avoidance/reduction of scars after surgical interventions on the human body and for local use in the postoperative treatment of ophthalmic disorders, e.g. the postoperative 30 treatment of glaucoma and in radiation-induced fibrosis or fibrosis induced by chemotherapy, in particular in the lung.
The compounds according to the invention are ester prodrugs of the corresponding carboxylic acids of the formula 1, in which B is a carboxyl 35 group.
.
CA 022~3282 1998-10-29 The compounds of the formula I are cleaved in the living body (in vivo) and in cell cultures (in vitro) to give compounds of the formula I in which B is a carboxyl group or its salts.
5 After the administration of the compounds of the formula 1, they cause the inhibition of collagen biosynthesis to be observed in vivo and in vitro, the compounds of the formula I in which B is a carboxyl group or its salts being formed. These compounds inhibit prolyl4-hydroxylase and therefore lead to an inhibition of collagen biosynthesis.
The compounds according to the invention correspond to the formula I
R2~oR4 (1) o 15 in which R1, R2 and R3 are hydrogen, R4 is (C1-C6)-alkyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is the radical of an alcohol GOH, including the physiologically active salts.
Preferred compounds of the formula I are those in which R1, R2 and R3 are hydrogen, R4 is (C1-C6)-alkyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is a branched or unbranched or cyclic (C1-C20)-alkyl radical, or a branched or unbranched or cyclic (C2-C20)-alkenyl radical, a CA 022~3282 1998-10-29 ~ WO 97/41103 PCT/EP96/01793 corresponding (C2-C20)-alkinyl radical, a corresponding (C4-C20)-alkeninyl radical or a retinyl radical, where the radicals can in each case contain one or more multiple bonds, or is a phenylalkyl radical, where the above radicals in particular contain one or more substituents selected from the group consisting of hydroxyl, halogen, cyano, trifluoromethyl, carboxyl, (C1-C12)-alkyl, (C3-C8)-cycloalkyl, (C5-C8)-cycloalkenyl, (C1-C12)-alkoxy, (C1-C12)-alkoxy-(C1-C12)-alkyl, (C1-C12)-alkoxy-(C1-C12)-alkoxy, phenyl-(C1-C4)-alkyloxy, (C1-C8)-hydroxyalkyl, (C1-C12)-alkylcarbonyloxy, (C3-C8)-cycloalkylcarbonyloxy, benzoyloxy, phenyl-(C1-C4)-alkylcarbonyloxy, including the physiologically active salts.
Preferred compounds of the formula I are furthermore those in which R1, R2 and R3 are hydrogen, R4 is (C1-C4)-alkyl, A is a -CH2 group, in which a hydrogen atom can be replaced by a methyl group, and B is -CO2G, where 20 G is a branched or unbranched or cyclic aliphatic (C1-C18)-alkyl radical, a (C3-C8)-cycloalkyl-(C1-C8)-alkyl radical, a branched or unbranched (C2-C18)-alkenyl radical, such as, for example, a geranyl, farnesyl or a retinyl radical, or a corresponding (C2-C18)-alkinyl radical, a benzyl, phenethyl, phenylpropyl or phenylbutyl radical, where the above radicals contain a substituent from the group consisting of hydroxyl, (C1-C4)-alkoxy, (C1-C6)-alkylcarbonyloxy, (C3-C8)-cycloalkylcarbonyloxy, benzoyloxy or phenyl-(C1-C4)-alkylcarbonyloxy, including the physiologically active salts.
Particularly preferred compounds of the formula I are those in which R1, R2 and R3 are hydrogen, 35 R4 is (C1-C4)-alkyl, A is a -CH2 group, in which a hydrogen atom can be replaced by a methyl group, and .. . ..
CA 022~3282 1998-10-29 B is -CO2G, where G is a branched or unbranched or cyclic aliphatic (C1-C18)-alkyl radical, a (C3-C8)-cycloalkyl-(C1-C4)-alkyl radical, a branched or unbranched (C2-C18)-alkenyl radical, a benzyl, phenethyl, phenylpropyl or phenylbutyl radical, including the physiologically active salts.
Particularly preferred compounds of the formula I are furthermore those in which R1, R2 and R3 are hydrogen, R4 is methyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and 15 B is-CO2G, where G is a branched or unbranched (C1-C18)-alkyl or (C2-C18)-alkenyl radical, including the physiologically active salts.
Particularly preferred compounds of the formula I are furthermore those in 20 which R1, R2 and R3 are hydrogen, ~ R4 is methyl, A is a -CH2 group and B is-CO2G, where G is a branched or unbranched (C1-C18)-alkyl or (C2-C18)-alkenyl radical, including the physiologically active salts.
30 In particular, preferred compounds of the formula I are those in which R1, R2 and R3 are hydrogen, R4 is methyl, 35 A is a -CH2 group, B is -CO2G, where G is a linear (C1-C18)-alkyl radical, . .
CA 022~3282 1998-10-29 including the physiologically active salts.
Particularly preferred compounds of the formula I are those in which 5 G is a linear (C1-C16)-alkyl radical, including the physiologically active salts.
The invention furthermore comprises salts of the compounds of the formula 1.
Salt formation with acidic reagents can be carried out at the pyridine N
atom.
Reagents used are, for example, toluenesulfonic acid, methanesulfonic 15 acid, HCI, H2SO4, H3PO4 and pharmaceuticals which contain an acidic group.
The invention relates to the compounds of the formula I and to the physiologically tolerable salts for use in the inhibition of collagen 20 biosynthesis, in particular in different cells.
The invention relates to the compounds of the formula I and to the physiologically tolerable salts for use in the inhibition of prolyl4-hydroxylase in vivo and in vitro.
The invention furthermore relates to the compounds of the formula I andthe physiologically tolerable salts for use in fibrotic disorders of the lung, the liver, the kidney, the heart, the eye and the skin. The compounds can also be used in atherosclerosis. In this connection, systemic and/or local 30 applications are used.
In particular, the invention relates to compounds of the formula I and to the physiologically tolerable salts for local use, in particular as inhibitors of collagen biosynthesis, as antifibrotic active compounds and in forms of 35 disease which are caused by an increased formation of connective tissue (collagen). These include the uses for the avoidance/reduction of scars after surgical interventions on the human body and in the postoperative treatment of eye operations, e.g. in glaucoma operations and in radiation-CA 022~3282 1998-10-29 induced fibrosis or fibrosis induced by chemotherapy, in particular in the lung.
Finally, the invention relates to the compounds of the formula I for use as pharmaceuticals.
In particular, the invention relates to compounds of the formula I for local use in fibrosis of the skin, the lung and the eye, in particular for the postoperative treatment of glaucoma.
The invention furthermore relates to a process for the preparation of compounds of the formula 1.
The compounds of the formula 1, in which A is a -CH2- group in which a hydrogen can be replaced by a methyl group and B is CO2G, are prepared by i1.) reacting pyridine-2-carboxylic acids of the formula ll (R5 = H) with the amino esters of the formula lll or their salts to give the amido esters of the formula l; or i2.) reacting pyridine-2-carboxylic acid esters of the formula ll (R5 = (C1-C16-alkyl) under the conditions of aminolysis to give the compounds of the formula l; or 25 ii) esterifying the compounds of the formula IV with an alcohol GOH, or iii) alkylating the compounds of the formula V with R4X, in which X is a leaving group, in particular halogen, -OSO2Me, -OSO2phenyl, etc.
CA 022~3282 1998-10-29 Scheme 1 R2~, oR4 H2N-A-C02G R2~" oR4 R C02R i2 ) R3 N ~NH-A-C02G
~ I ~
R ~ R X iii ) R2 ~N H-A-C02H R2 ~,OH
R3 N o R3 --~NH-A-C02G
IV V ~
5 Suitable processes for amide formation (reaction i1 ) are the methods of carbonyl activation and the condensation reactions known from peptide chemistry.
Reagents used for carboxylic acid activation can be the substances known 10 to the person skilled in the art, such as thionyl chloride, oxalyl chloride, pivaloyl chloride, chloroformic acid ester derivatives or N,N'-carbonyldimidazole. The activated derivatives of the compounds of the formula ll are reacted in situ with the amide derivatives of the formula lll after preparation.
A suitable condensing agent is, for example, the combination of N, N'-dicyclohexylcarbodiimide/N-hydroxy-1 H-benzotriazole and N-ethylmorpholine.
20 Suitable solvents are dichloromethane, tetrachloromethane, butyl acetate, ethyl acetate, toluene, tetrahydrofuran, dimethoxyethane, 1,4-dioxane, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, nitromethane and/or pyridine.
....... ..
CA 022~3282 1998-10-29 The compounds of the formula I according to the invention have valuablepharmacological properties and exhibit, in particular, antifibrotic activity.
The antifibrotic action can be determined in the following animal models:
formation of a connective-tissue capsule around a subcutaneously implanted osmotic minipump, in: Unemori, E.N. et al., J. Invest. Dermatol., 101: 280-5,1993, 10 collagen content of subcutaneously implanted "cotton pellets" or polyvinyl sponges, in: Boyle, E., Mangan, F.R., Br. J. Ep. Pathnol. 61: 351-60,1980;
and collagen content of the lung after radiation-induced or chemically induced 15 fibrosis, in: 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 cultures:
normal human dermal fibroblasts (NHDF), rat liver epithelial cells (Ref.1) 20 and fat storing cells from the liver (Ref. 2) are used as a cell model for substance testing. To do this, the cells are cultured in the presence of inhibitors. Simultaneously, the collagen synthesized de novo this time is metabolically labeled by 4-3H-L-proline and 14C-proline. The effect of the test substances on the degree of hydroxylation of the collagen is then 25 determined according to the method of Chojkier et al. (Ref. 3). The reference substance employed 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: 3541, 2.: Blomhoff, R., Berg T.1990. Isolation and cultivation of 30 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 [4-3H]:[14C]proline in collagenase digests. Anal. Biochem. 108: 385-393).
35 Inhibition of prolyl-4-hydroxylase in vitro (in chicken embryo calvaria) according to Biochem. J. (1994) 300, 525-300:
CA 022~3282 1998-10-29 1. Metabolic labeling Calvaria are dissected from 15-day-old chicken embryos and washed for 3 min at 37~C in Hanks' balanced salt solution minimum essential medium Eagle (HMEM, BioWhittaker, Walkersville, MD, USA). 4 calvaria each are 5 incubated at 37~C for 2.5 h in glass vessels with 1.5 ml of HMEM with addition of 2 mM glutamine and 1 ~Ci [lacuna] [U-14C]proline and various inhibitor concentrations. The incubation is ended by inserting the sample vessels in ice. The medium is removed and the calvaria are briefly washed with 1 ml of double-distilled H2O. The calvaria are then treated with 3 ml of 10 0.5 M acetic acid and 18 1l9 of phenylmethanesulfonyl fluoride and extracted for 16 h. The extraction solution is dialyzed against 0.5 M acetic acid at 4~C to remove the free [U-14C]proline and then freeze-dried in aliquots.
15 2. Hydroxyproline - analysis An aliquot of the freeze-dried sample is dissolved in 2 ml of 6 N HCI and hydrolyzed at 105~C for 24 h. The hydrochloric acid is then evaporated.
The residue is resuspended in 300 1ll of double-distilled H20, transferred to Eppendorf vessels and dried again. The residual hydrochlorides are 20 neutralized by dissolving in 60 ~11 of an ethanol/H20/triethylamine 2:2:1 (v:v:v) [lacuna] and repeated drying. The amino acids are then derivatized for 20 min at room temperature in a solution of ethanol/triethylamine/phenyl isothiocyanate/H2O 7:1:1:1 (v:v:v:v) and dried again. For analysis using HPLC, the samples are dissolved in 150 ~11 of a phosphate buffer (5 mM
25 Na2HPO4, pH 7.4/acetonitrile 95:5 (v:v)) and centrifuged at 10,000 9 for 5 min. 50 ~11 are used for analysis. The HPLC chromatography is carried out on a C18 reverse phase column Ultrasphere ODS 3 llm, 4.6 nim x 7.5 cm (Beckman) at 50~C using the following gradient system:
Time Buffer A Buffer B
0 min 100%
0-9 min 100-90% 0-10%
9-11 min 90-0% 10-100%
11-12 min 0% 100%
12-14 min 0-100% 100-0%
1 4-19 min 100%
CA 022~3282 1998-10-29 Soln. A: 70 mM Na acetate pH 6.14/0.1% acetonitrile Soln. B: acetonitrile/methanol/water45:15:40 (v:v:v) 3. SDS - polyacrylamide - gel electrophoresis An aliquot of the freeze-dried samples from 1. is dissolved for the SDS
polyacrylamide - gel electrophoresis using 10 mM tris/HCI, pH 8/1 mM
EDTA/1 % SDS and 5% mercaptoethanol and denatured at 95~C for 5 min.
The electrophoresis is carried out on a linear gradient gel (5-15%). After 10 fixing in methanol/glacial acetic acid/water 3:1:6 (v:v:v), the gels are dried and used for the exposure of Kodak X-Omat film at -70~C.
In the following, the surprising advantages of the compounds according to the invention compared with compounds from EP-A-0 650 960 are 15 illustrated with the aid of comparison experiments by means of the inhibition of prolyl4-hydroxylase (collagen biosynthesis) in rat liver cells:
Table 1:
Compounds from Inhibition of collagen biosynthesis in Example No. rat liver cells of EP-A-0 650 960 rat liver fat storing cells 27 -79%: 100,um 151 -65%: 25 llm 154 40%: 25,um 155 -59%: 50 ,um 239 41%: 25 ~m of EP-A-0 650 960 rat liver epithelial cells 290 -86%: 25 ~lm 288 -72%: 25 ~lm CA 022~3282 1998-10-29 Table 2:
Compounds from Inhibition of collagen biosynthesis in Example No. rat liver cells of the present application rat liver fast storing cells 2 -50%: 1 .5 + 0. 1 ~m 3 -50%: 0.9 + 0.1 ~m 14 -50%: 10.0 ~lm of the present application rat liver epithelial cells 2 -50%: 7,um 14 -50%: 10 llm Table 3:
Compounds from Example No. in calvaria of the present application 2 -50%: 0.032 llm 14 -50%: 0.41 ~lm in normal human fibrobla .ts (NHDF) -50%: 0.1 ,um 2 -50%: 0.2 llm 3 -50%: 0.5 ~lm The comparisons show that compounds according to the invention of the present selection invention, compared to those of EP-A-0 650 960, exhibit 10 high inhibition rates (-50%) even at surprisingly low concentrations (s 10 ~lm, depending on the experimental model).
The compounds of the formula I can be used as medicaments in the form of pharmaceutical preparations which contain them, optionally with 15 tolerable pharmaceutical vehicles. The compounds can be used as therapeutics, e.g. in the form of pharmaceutical preparations, which contain these compounds as a mixture with a pharmaceutical, organic or inorganic excipient suitable for enteral, percutaneous or parenteral administration, or administration by inhalation, such as, for example, water, CA 022~3282 1998-10-29 gum arabic, gelatin, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, petroleum jelly etc.
For this purpose, they can be administered orally in doses from 0.1 to 25 5 mg/kg/day, preferably 1 to 5 mg/kg/day, or parenterally in doses of 0.01 to 5 mg/kg/day, preferably 0.01 to 2.5 mg/kg/day, in particular 0.5 to 1.0 mg/kg/day. In severe cases, the dose can also be increased. In many cases, however, lower doses are sufficient. These details relate to an adult approximately 75 kg in weight.
3-Methoxypyridine-2-carboxylic acid is known from J. Org. Chem. 36, 2002 (1971) by E.V. Brown, M.B. Shambhu (cf. formula ll; R1-R3 = H, R4 = Me, R5 = H).
15 Furthermore, methyl 3-methoxypyridine-2-carboxylate (cf. formula ll, R5 =
CH3) is known from Acta Chem. Scand. 23, 1791 (1969).
3-Methoxypyridine-2-carboxylic acid, which was used as starting material in the examples below, was obtained starting from 4-chloro-3-methoxy-2-20 methylpyridine, which was obtained from maltol (cf. EP-A-0 208 452 and EP-A-0 304 732).
Under the examples described below, the compounds of the formula I
according to the invention are designated as substituted pyridine-2-25 carboxylic acid (glycyl ester)amides.
This manner of notation is understood as meaning substituted pyridine-2-carboxylic acid N-((alkoxycarbonyl)methyl)amides.
Classification as substituted N-(pyridyl-2-carbonyl)glycines is a further possibility.
Example 1 N-(((1 -Butyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide a) 3-Methoxypyridine-2-carboxylic acid hydrochloride a1.) 4-Chloro-3-methoxy-2-methylpyridine 1-oxide 11.2 9 (80.5 mmol) of 3-methoxy-2-methyl4(1H)-pyridone in 100 ml of phosphorus oxychloride were heated under reflux for 10 hours.
CA 022~3282 1998-10-29 The mixture was subsequently concentrated, admixed 2 ml with 30 ml of toluene each time and concentrated again, and the residue was taken up in 150 ml of water, adjusted to pH 11 using K2CO3 and extracted with dichloromethane, and the organic phase was washed with water, dried and freed of solvent. The light-brown oil (9 g) gave, using m-chloroperbenzoic acid in dichloromethane under standard conditions, 8 g of the product, m.p. 88 to 89~C (from petroleum ether).
10 a2.) 4-Chloro-2-hydroxymethyl-3-methoxypyridine 30 g (173 mmol) of 4-chloro-3-methoxy-2-methylpyridine N-oxide were dissolved in 100 ml of glacial acetic acid and admixed dropwise with stirring at 80~C with 150 ml of acetic anhydride, and the mixture was stirred at 110~C for 2 hours. The mixture was then cooled to 80~C, 200 ml of methanol were added dropwise, the mixture was heated to the boil for 15 minutes and, after cooling, concentrated under reduced pressure and the residue was taken up in methanol and poured into 300 ml of 1.5 N methanolic sodium hydroxide solution, stirred at 20~C for 30 minutes and concentrated under reduced pressure, the residue was taken up in water and extracted three times with dichloromethane, the organic phase was dried and concentrated and the residue was crystallized with petroleum ether. This gave 23 g of product, m.p. 64 to 66~C.
25 a3.) 4-Chloro-3-methoxypyridine-2-carboxylic acid At 20~C, 52.2 g (0.3 mol) of the above alcohol were added to 24 g of KOH dissolved in 900 ml of water, and the mixture was admixed a little at a time, in each case until discoloration, with 70 g (0.44 mol) of potassium permanganate at 50 to 60~C. After 1 hour of stirring at 50~C, the hot mixture was filtered off with suction, the filter cake was rinsed three times with hot water and the filtrate was concenllated under reduced pressure to 300 ml and, with cooling, adjusted to pH
1 using concentrated HCI. Filtration with suction and drying gave 50 g of product, m.p., 121~C (n. decomp.).
a4.) 29 g (0.15 mol) of the above 4-chloropyridinecarboxylic acid in 500 ml of methanol/tetrahydrofuran (1:1) were hydrogenated in a hydrogenation apparatus with Pd/C (10%). After 3.1 l of hydrogen CA 022~3282 1998-10-29 ~O 97141103 PCT/EP96/01793 had been taken up, the catalyst was removed by filtration with suction, the filtrate was concentrated and the crystalline residue was treated with ethyl acetate, filtered offwith suction and dried; yield 29 9, m.p. 170~C (with decomp.).
b) 3.8 9 (20 mmol) of 3-methoxypyridine-2-carboxylic acid hydrochloride were suspended in 500 ml of anhydrous dichloromethane and, at 20~C and with stirring, admixed with 6.1 9 (20 mmol) of glycine 1-butyl ester tosylate (prepared from glycine, 1-butanol, p-toluenesulfonic acid on a water separator with toluene), then with 7.5 ml (60 mmol) of N-ethylmorpholine, 3 9 (22.5 mmol) of 1-hydroxy-1H-benzotriazole and 8.5 9 (20 mmol) of N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide metho-p-toluenesulfonate (CMC) and stirred at 20~C for 24 hours. Undissolved particles were then filtered off with suction and the filtrate was extracted successively with aqueous Na bicarbonate solution, with 1N aqueous hydrochloric acid and with water. The organic phase was dried and concentrated under reduced pressure and the oily residue was crystallized by scratching. This gave 1.76 9 of the colorless title compound, m.p. 60 to 62~C.
Example 2 N-(((1 -Octyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide 25 2.5 9 (13 mmol) of 3-methoxypyridine-2-carboxylic acid hydrochloride, 5.5 ml (45 mmol) of N-ethylmorpholine, 2 9 (15 mmol) of 1-hydroxy-1H-benzotriazole, 4.7 9 (13 mmol) of glycine octyl ester tosylate (prepared on a water separator with toluene from glycine, 1-octanol and p-toluenesulfonic acid) and 6.3 9 (15 mmol) of CMC (cf. Example 1) were 30 stirred in 350 [lacuna] of anhydrous dichloromethane for 48 hours. After work-up by the method of Example 1), the crude product was chromatographed over silica gel using dichloromethane (with an addition of up to 2.5% of methanol in the course of the elution). This gave 3.6 9 of the colorless oily title compound, 1H NMR (CDCI3): ~ = 4.26 (d, CH2-glycine).
CA 022~3282 1998-10-29 Example 3 N-(((1 -Dodecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide By the method of Example 1, 2.8 g (15 mmol) of 3-methoxypyridine-2-5 carboxylic acid hydrochloride in 400 ml of anhydrous dichloromethane were stirred at 20~C with 6.2 9 (15 mmol) of glycine 1-dodecyl ester tosylate (prepared on a water separator from 38 g (0.5 mol) of glycine, 93.2 9 (0.5 mol) of 1-dodecanol and 115 9 (0.6 mol) of p-toluenesulfonic acid; 140 9 of product were crystallized with diethyl ether, m.p. about 106~C, sintered at 80~C), 8.2 ml (60 mmol) of N-ethylmorpholine, 2 9 (15 mmol) of 1-hydroxy-1H-benzotriazole and 6.3 9 (15 mmol) of CMC for 24 hours. Owing to a strong emulsion formation, for work-up, similarly to Example 1, dichloromethane was distilled off under reduced pressure and the residue was taken up in diethyl ether. The organic phase was dried 15 with magnesium sulfate and concentrated under reduced pressure, and 4.7 9 of the resulting yellow oil were then chromatographed over silica gel using ethyl acetate. This gave 4 9 of a light-yellow oil which crystallized after standing for 2 days after scratching with a glass rod. 3.6 9 of the colorless title compound, m.p. 47 to 49~C, were isolated.
Example 4 N-(((1 -Pentyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 5 25 N-(((1-Hexyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 6 N-(((1 -Heptyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
30 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.
CA 022~3282 1998-10-29 Example 10 N-(((1 -Octadecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 11 5 N-(((1-Propyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 12 N-(((2-Propyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
10 Example 13 N-(((1 -Tridecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 14 N-(((1 -Hexadecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
5.7 9 (30 mmol) of 4-chloro-3-methoxypyridine-2-carboxylic acid and 14.2 g (30 mmol) of glycine hexadecyl ester tosylate (m.p. about 90~C, prepared from glycine,1-hexadecanol, p-toluenesulfonic acid on a water separator using toluene) in 300 ml of dichloromethane were admixed by the method 20 of Example 1) with 7.7 [lacuna] (60 mmol) of N-ethylmorpholine, 4.5 g (33 mmol) of 1-hydroxy-1 H-benzotriazole and 12.8 9 (30 mmol) of N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide metho-p-toluenesulfonate (CMC) and stirred for 24 hours. Undissolved particles were then filtered off with suction and the filtrate was extracted with aqueous Na bicarbonate 25 solution, with water and with aqueous hydrochloric acid, the organic phase was concentrated and the residue (14 9) was dissolved in 500 ml of tetrahydrofuran/methanol (1:1), admixed with Pd/C (10%) and hydrogenated in a hydrogenation apparatus.
After the hydrogen uptake had ended, the catalyst was filtered off with 30 suction, the filtrate was concentrated and the residue was chromatographed over silica gel using ethyl acetate. The appropriate fractions were concentrated and the residue was crystallized using diisopropyl ether. This gave 2.1 9 of the title compound as a colorless substance, m.p. 63 to 65~C.
CA 022~32X2 1998-10-29 Example 15 N-((Ethyloxycarbonyl)methyl)-3-methoxypyridine-2-carboxamide hydrochloride 5 a) N-((Ethyloxycarbonyl)methyl)-4-chloro-3-methoxypyridine-2-carboxamide For the preparation, 4.7 9 (25 mmol) of 4-chloro-3-methoxypyridine-2-carboxylic acid were suspended in 200 ml of anhydrous dichloromethane and, at 20~C and with stirring, admixed successively with 3.5 9 (25 mmol) of glycine ethyl ester hydrochloride, 6.4 ml (50 mmol) of N-ethylmorpholine, 3.8 9 (28 mmol) of 1-hydroxy-(1 H)-benzotriazole and 5.15 9 (25 mmol) of N,N'-dicyclohexylcarbodiimide and stirred at 20~C for 20 hours.
Undissolved particles were then filtered off, the organic phase was shaken with saturated aqueous sodium carbonate solution, dried and concentrated under reduced pressure and the residue (6 9 of oil) was chromatographed over silica gel using ethyl acetate, giving 5.4 9 of an oily product.
20 B) The title compound was obtained by hydrogenating the above substance with Pd/C (10%) in a hydrogenation apparatus, m.p. 141 - to 142~C (with evolution of gas, from diethyl ether).
Example 16 25 N-(((2-Nonyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide racemate .
2.5 9 (10 mmol) of N-(carboxymethyl)-3-methoxypyridine-2-carboxamide hydrochloride (m.p. 157~C, with evolution of gas) were suspended in 30 100 ml of anhydrous tetrahydrofuran and admixed with 1.6 ml (12 mmol) of triethylamine and then dropwise with stirring with 2.4 9 of pivaloyl chloride dissolved in a little tetrahydrofuran (the temperature rises to 35-40~C).
After 30 min, the mixture was concentrated under reduced pressure and the reddish residue was taken up in 100 ml of anhydrous tetrahydrofuran 35 and admixed with 1.6 ml of triethylamine, and the mixture was then added at 20~C to a solution of Na 2-nonanolate in 2-nonanol (prepared from 30 ml of 2-nonanol and 0.8 9 (20 mmol) of NaH). After 1 h, the mixture was concentrated under reduced pressure, the residue was admixed with .. .. . ..
CA 022~32X2 1998-10-29 dichloromethane and extracted with 2N of aqueous ammonium chloride solution, the organic phase was dried and concentrated under reduced pressure and the residue (9 g) was chromatographed over silica gel using ethyl acetate. This gave 1.1 9 of the oily colorless title compound, 5 1H NMR (DMSO): ~ = 3.95 (d, CH2-glycine).
Example 17 N-(((4-Heptyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide 10 2.5 g (10 mmol) of N-(carboxymethyl)-3-methoxypyridine-2-carboxamide hydrochloride were treated as in Example 16 and then admixed at 20~C
with 140 ml of a solution of Na 4-heptanolate in 4-heptanol (prepared from 140 ml of 4-heptanol and 0.6 g (25 mmol) of sodium, ultrasonic bath). After 30 minutes, the mixture was heated at 70 to 80~C for 1 hour and, after -15 cooling, concentrated under reduced pressure, the residue was taken up inwater and extracted with dichloromethane and the organic phase was concentrated under reduced pressure and dried at an oil pump. The oily crude product crystallized after approximately 15 hours, m.p. 75 to 78~C.
20 Example 18 N-(((3-Pentyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide Example 19 N-(((1 -(cis-9-Octadecenyl)oxy)carbonyl)methyl)-3-methoxypyridine-2-25 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 35 Example 22 N-((Methyloxycarbonyl)methyl)-3-methoxypyridine-2-carboxamide . .
Compounds which inhibit the enzymes prolyl and Iysyl hydroxylase cause a very selective inhibition of collagen biosynthesis by affecting the collagen-10 specific hydroxylation reactions. In the course thereof, protein-bound proline or Iysine is hydroxylated by the enzymes prolyl or Iysyl hydroxylase.
If this reaction is suppressed by inhibitors, a nonfunctional, underhydroxylated collagen molecule results, which can be released into the extracellular space only in a small amount by the cells. The 15 underhydroxylated collagen additionally cannot be incorporated into the collagen matrix and is very easily proteolytically degraded. As a result of these effects, the amount of extracellularly deposited collagen is decreased overall.
20 Inhibitors of prolyl hydroxylase are therefore suitable substances in the therapy of disorders in which the deposition of collagens decisively contributes to the clinical picture. These include, inter alia, fibroses of the lung, liver and skin (scleroderma and scars after burns, injuries and surgical interventions) and also atherosclerosis.
It is known that the enzyme prolyl hydroxylase is effectively inhibited by pyridine-2,4- and -2,5-dicarboxylic acid (K. Majamaa et al., Eur. J.
Biochem.138 (1984) 239-245). In cell culture, however, these compounds are only active as inhibitors in very high concentrations (Tschank, G. et al., 30 Biochem. J. 238 (1987) 625-633).
Prodrugs of the pyridine-2,4(5)-dicarboxylates are also known. These are described in the Patent Applications EP-A-0 590 520 and DE-A42 38 506 and EP-A-0 562 512.
35 N-Oxalylglycines as inhibitors of prolyl-4-hydroxylase are known from J.Med.Chem.1992, 35, 2652-2658 (Cunliffe et al.) and EP-A-0 457 163.
~ . . . .
CA 022~3282 1998-10-29 Furthermore, 3-hydroxypyridine-2-carboxylic acid (glycyl ethyl ester)amide is described in J. Org. Chem. 31, 636-638 (1966).
It was therefore the object to provide compounds which are distinguished 5 by a particularly high in vivo and/or in vitro activity, in particular in their systemic and/or local use.
Surprisingly, it has now been found that 3-alkoxypyridine-2-carboxamidoesters of the formula I are particularly strongly active inhibitors 10 of collagen biosynthesis. The compounds of the invention are selected from the compounds described in EP-A-0 650 960. Compared with the compounds mentioned in the European Patent Application, they are distinguished by a particularly high in vivo and in vitro activity, in particular in their systemic and/or local use against fibrotic disorders. These include, 15 for example, fibroses of the lung, the liver, the kidney, the heart, the eye and the skin and in atherosclerosis.
The compounds of the formula l lead to a potent inhibition of collagen biosynthesis in the most diverse cells (e.g. normal human dermal 20 fibroblasts, primary fat storing cells from rat liver, rat liver epithelial cells and in organ cultures of calvaria).
They are particularly suitable for local use as inhibitors of collagen biosynthesis, for local use as antifibrotic active compounds and for local 25 use in forms of disease which are caused by an increased formation of connective tissue (collagen). Thus the compounds of the formula I are suitable, for example, for local use for the avoidance/reduction of scars after surgical interventions on the human body and for local use in the postoperative treatment of ophthalmic disorders, e.g. the postoperative 30 treatment of glaucoma and in radiation-induced fibrosis or fibrosis induced by chemotherapy, in particular in the lung.
The compounds according to the invention are ester prodrugs of the corresponding carboxylic acids of the formula 1, in which B is a carboxyl 35 group.
.
CA 022~3282 1998-10-29 The compounds of the formula I are cleaved in the living body (in vivo) and in cell cultures (in vitro) to give compounds of the formula I in which B is a carboxyl group or its salts.
5 After the administration of the compounds of the formula 1, they cause the inhibition of collagen biosynthesis to be observed in vivo and in vitro, the compounds of the formula I in which B is a carboxyl group or its salts being formed. These compounds inhibit prolyl4-hydroxylase and therefore lead to an inhibition of collagen biosynthesis.
The compounds according to the invention correspond to the formula I
R2~oR4 (1) o 15 in which R1, R2 and R3 are hydrogen, R4 is (C1-C6)-alkyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is the radical of an alcohol GOH, including the physiologically active salts.
Preferred compounds of the formula I are those in which R1, R2 and R3 are hydrogen, R4 is (C1-C6)-alkyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is a branched or unbranched or cyclic (C1-C20)-alkyl radical, or a branched or unbranched or cyclic (C2-C20)-alkenyl radical, a CA 022~3282 1998-10-29 ~ WO 97/41103 PCT/EP96/01793 corresponding (C2-C20)-alkinyl radical, a corresponding (C4-C20)-alkeninyl radical or a retinyl radical, where the radicals can in each case contain one or more multiple bonds, or is a phenylalkyl radical, where the above radicals in particular contain one or more substituents selected from the group consisting of hydroxyl, halogen, cyano, trifluoromethyl, carboxyl, (C1-C12)-alkyl, (C3-C8)-cycloalkyl, (C5-C8)-cycloalkenyl, (C1-C12)-alkoxy, (C1-C12)-alkoxy-(C1-C12)-alkyl, (C1-C12)-alkoxy-(C1-C12)-alkoxy, phenyl-(C1-C4)-alkyloxy, (C1-C8)-hydroxyalkyl, (C1-C12)-alkylcarbonyloxy, (C3-C8)-cycloalkylcarbonyloxy, benzoyloxy, phenyl-(C1-C4)-alkylcarbonyloxy, including the physiologically active salts.
Preferred compounds of the formula I are furthermore those in which R1, R2 and R3 are hydrogen, R4 is (C1-C4)-alkyl, A is a -CH2 group, in which a hydrogen atom can be replaced by a methyl group, and B is -CO2G, where 20 G is a branched or unbranched or cyclic aliphatic (C1-C18)-alkyl radical, a (C3-C8)-cycloalkyl-(C1-C8)-alkyl radical, a branched or unbranched (C2-C18)-alkenyl radical, such as, for example, a geranyl, farnesyl or a retinyl radical, or a corresponding (C2-C18)-alkinyl radical, a benzyl, phenethyl, phenylpropyl or phenylbutyl radical, where the above radicals contain a substituent from the group consisting of hydroxyl, (C1-C4)-alkoxy, (C1-C6)-alkylcarbonyloxy, (C3-C8)-cycloalkylcarbonyloxy, benzoyloxy or phenyl-(C1-C4)-alkylcarbonyloxy, including the physiologically active salts.
Particularly preferred compounds of the formula I are those in which R1, R2 and R3 are hydrogen, 35 R4 is (C1-C4)-alkyl, A is a -CH2 group, in which a hydrogen atom can be replaced by a methyl group, and .. . ..
CA 022~3282 1998-10-29 B is -CO2G, where G is a branched or unbranched or cyclic aliphatic (C1-C18)-alkyl radical, a (C3-C8)-cycloalkyl-(C1-C4)-alkyl radical, a branched or unbranched (C2-C18)-alkenyl radical, a benzyl, phenethyl, phenylpropyl or phenylbutyl radical, including the physiologically active salts.
Particularly preferred compounds of the formula I are furthermore those in which R1, R2 and R3 are hydrogen, R4 is methyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and 15 B is-CO2G, where G is a branched or unbranched (C1-C18)-alkyl or (C2-C18)-alkenyl radical, including the physiologically active salts.
Particularly preferred compounds of the formula I are furthermore those in 20 which R1, R2 and R3 are hydrogen, ~ R4 is methyl, A is a -CH2 group and B is-CO2G, where G is a branched or unbranched (C1-C18)-alkyl or (C2-C18)-alkenyl radical, including the physiologically active salts.
30 In particular, preferred compounds of the formula I are those in which R1, R2 and R3 are hydrogen, R4 is methyl, 35 A is a -CH2 group, B is -CO2G, where G is a linear (C1-C18)-alkyl radical, . .
CA 022~3282 1998-10-29 including the physiologically active salts.
Particularly preferred compounds of the formula I are those in which 5 G is a linear (C1-C16)-alkyl radical, including the physiologically active salts.
The invention furthermore comprises salts of the compounds of the formula 1.
Salt formation with acidic reagents can be carried out at the pyridine N
atom.
Reagents used are, for example, toluenesulfonic acid, methanesulfonic 15 acid, HCI, H2SO4, H3PO4 and pharmaceuticals which contain an acidic group.
The invention relates to the compounds of the formula I and to the physiologically tolerable salts for use in the inhibition of collagen 20 biosynthesis, in particular in different cells.
The invention relates to the compounds of the formula I and to the physiologically tolerable salts for use in the inhibition of prolyl4-hydroxylase in vivo and in vitro.
The invention furthermore relates to the compounds of the formula I andthe physiologically tolerable salts for use in fibrotic disorders of the lung, the liver, the kidney, the heart, the eye and the skin. The compounds can also be used in atherosclerosis. In this connection, systemic and/or local 30 applications are used.
In particular, the invention relates to compounds of the formula I and to the physiologically tolerable salts for local use, in particular as inhibitors of collagen biosynthesis, as antifibrotic active compounds and in forms of 35 disease which are caused by an increased formation of connective tissue (collagen). These include the uses for the avoidance/reduction of scars after surgical interventions on the human body and in the postoperative treatment of eye operations, e.g. in glaucoma operations and in radiation-CA 022~3282 1998-10-29 induced fibrosis or fibrosis induced by chemotherapy, in particular in the lung.
Finally, the invention relates to the compounds of the formula I for use as pharmaceuticals.
In particular, the invention relates to compounds of the formula I for local use in fibrosis of the skin, the lung and the eye, in particular for the postoperative treatment of glaucoma.
The invention furthermore relates to a process for the preparation of compounds of the formula 1.
The compounds of the formula 1, in which A is a -CH2- group in which a hydrogen can be replaced by a methyl group and B is CO2G, are prepared by i1.) reacting pyridine-2-carboxylic acids of the formula ll (R5 = H) with the amino esters of the formula lll or their salts to give the amido esters of the formula l; or i2.) reacting pyridine-2-carboxylic acid esters of the formula ll (R5 = (C1-C16-alkyl) under the conditions of aminolysis to give the compounds of the formula l; or 25 ii) esterifying the compounds of the formula IV with an alcohol GOH, or iii) alkylating the compounds of the formula V with R4X, in which X is a leaving group, in particular halogen, -OSO2Me, -OSO2phenyl, etc.
CA 022~3282 1998-10-29 Scheme 1 R2~, oR4 H2N-A-C02G R2~" oR4 R C02R i2 ) R3 N ~NH-A-C02G
~ I ~
R ~ R X iii ) R2 ~N H-A-C02H R2 ~,OH
R3 N o R3 --~NH-A-C02G
IV V ~
5 Suitable processes for amide formation (reaction i1 ) are the methods of carbonyl activation and the condensation reactions known from peptide chemistry.
Reagents used for carboxylic acid activation can be the substances known 10 to the person skilled in the art, such as thionyl chloride, oxalyl chloride, pivaloyl chloride, chloroformic acid ester derivatives or N,N'-carbonyldimidazole. The activated derivatives of the compounds of the formula ll are reacted in situ with the amide derivatives of the formula lll after preparation.
A suitable condensing agent is, for example, the combination of N, N'-dicyclohexylcarbodiimide/N-hydroxy-1 H-benzotriazole and N-ethylmorpholine.
20 Suitable solvents are dichloromethane, tetrachloromethane, butyl acetate, ethyl acetate, toluene, tetrahydrofuran, dimethoxyethane, 1,4-dioxane, acetonitrile, N,N-dimethylformamide, N,N-dimethylacetamide, dimethyl sulfoxide, nitromethane and/or pyridine.
....... ..
CA 022~3282 1998-10-29 The compounds of the formula I according to the invention have valuablepharmacological properties and exhibit, in particular, antifibrotic activity.
The antifibrotic action can be determined in the following animal models:
formation of a connective-tissue capsule around a subcutaneously implanted osmotic minipump, in: Unemori, E.N. et al., J. Invest. Dermatol., 101: 280-5,1993, 10 collagen content of subcutaneously implanted "cotton pellets" or polyvinyl sponges, in: Boyle, E., Mangan, F.R., Br. J. Ep. Pathnol. 61: 351-60,1980;
and collagen content of the lung after radiation-induced or chemically induced 15 fibrosis, in: 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 cultures:
normal human dermal fibroblasts (NHDF), rat liver epithelial cells (Ref.1) 20 and fat storing cells from the liver (Ref. 2) are used as a cell model for substance testing. To do this, the cells are cultured in the presence of inhibitors. Simultaneously, the collagen synthesized de novo this time is metabolically labeled by 4-3H-L-proline and 14C-proline. The effect of the test substances on the degree of hydroxylation of the collagen is then 25 determined according to the method of Chojkier et al. (Ref. 3). The reference substance employed 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: 3541, 2.: Blomhoff, R., Berg T.1990. Isolation and cultivation of 30 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 [4-3H]:[14C]proline in collagenase digests. Anal. Biochem. 108: 385-393).
35 Inhibition of prolyl-4-hydroxylase in vitro (in chicken embryo calvaria) according to Biochem. J. (1994) 300, 525-300:
CA 022~3282 1998-10-29 1. Metabolic labeling Calvaria are dissected from 15-day-old chicken embryos and washed for 3 min at 37~C in Hanks' balanced salt solution minimum essential medium Eagle (HMEM, BioWhittaker, Walkersville, MD, USA). 4 calvaria each are 5 incubated at 37~C for 2.5 h in glass vessels with 1.5 ml of HMEM with addition of 2 mM glutamine and 1 ~Ci [lacuna] [U-14C]proline and various inhibitor concentrations. The incubation is ended by inserting the sample vessels in ice. The medium is removed and the calvaria are briefly washed with 1 ml of double-distilled H2O. The calvaria are then treated with 3 ml of 10 0.5 M acetic acid and 18 1l9 of phenylmethanesulfonyl fluoride and extracted for 16 h. The extraction solution is dialyzed against 0.5 M acetic acid at 4~C to remove the free [U-14C]proline and then freeze-dried in aliquots.
15 2. Hydroxyproline - analysis An aliquot of the freeze-dried sample is dissolved in 2 ml of 6 N HCI and hydrolyzed at 105~C for 24 h. The hydrochloric acid is then evaporated.
The residue is resuspended in 300 1ll of double-distilled H20, transferred to Eppendorf vessels and dried again. The residual hydrochlorides are 20 neutralized by dissolving in 60 ~11 of an ethanol/H20/triethylamine 2:2:1 (v:v:v) [lacuna] and repeated drying. The amino acids are then derivatized for 20 min at room temperature in a solution of ethanol/triethylamine/phenyl isothiocyanate/H2O 7:1:1:1 (v:v:v:v) and dried again. For analysis using HPLC, the samples are dissolved in 150 ~11 of a phosphate buffer (5 mM
25 Na2HPO4, pH 7.4/acetonitrile 95:5 (v:v)) and centrifuged at 10,000 9 for 5 min. 50 ~11 are used for analysis. The HPLC chromatography is carried out on a C18 reverse phase column Ultrasphere ODS 3 llm, 4.6 nim x 7.5 cm (Beckman) at 50~C using the following gradient system:
Time Buffer A Buffer B
0 min 100%
0-9 min 100-90% 0-10%
9-11 min 90-0% 10-100%
11-12 min 0% 100%
12-14 min 0-100% 100-0%
1 4-19 min 100%
CA 022~3282 1998-10-29 Soln. A: 70 mM Na acetate pH 6.14/0.1% acetonitrile Soln. B: acetonitrile/methanol/water45:15:40 (v:v:v) 3. SDS - polyacrylamide - gel electrophoresis An aliquot of the freeze-dried samples from 1. is dissolved for the SDS
polyacrylamide - gel electrophoresis using 10 mM tris/HCI, pH 8/1 mM
EDTA/1 % SDS and 5% mercaptoethanol and denatured at 95~C for 5 min.
The electrophoresis is carried out on a linear gradient gel (5-15%). After 10 fixing in methanol/glacial acetic acid/water 3:1:6 (v:v:v), the gels are dried and used for the exposure of Kodak X-Omat film at -70~C.
In the following, the surprising advantages of the compounds according to the invention compared with compounds from EP-A-0 650 960 are 15 illustrated with the aid of comparison experiments by means of the inhibition of prolyl4-hydroxylase (collagen biosynthesis) in rat liver cells:
Table 1:
Compounds from Inhibition of collagen biosynthesis in Example No. rat liver cells of EP-A-0 650 960 rat liver fat storing cells 27 -79%: 100,um 151 -65%: 25 llm 154 40%: 25,um 155 -59%: 50 ,um 239 41%: 25 ~m of EP-A-0 650 960 rat liver epithelial cells 290 -86%: 25 ~lm 288 -72%: 25 ~lm CA 022~3282 1998-10-29 Table 2:
Compounds from Inhibition of collagen biosynthesis in Example No. rat liver cells of the present application rat liver fast storing cells 2 -50%: 1 .5 + 0. 1 ~m 3 -50%: 0.9 + 0.1 ~m 14 -50%: 10.0 ~lm of the present application rat liver epithelial cells 2 -50%: 7,um 14 -50%: 10 llm Table 3:
Compounds from Example No. in calvaria of the present application 2 -50%: 0.032 llm 14 -50%: 0.41 ~lm in normal human fibrobla .ts (NHDF) -50%: 0.1 ,um 2 -50%: 0.2 llm 3 -50%: 0.5 ~lm The comparisons show that compounds according to the invention of the present selection invention, compared to those of EP-A-0 650 960, exhibit 10 high inhibition rates (-50%) even at surprisingly low concentrations (s 10 ~lm, depending on the experimental model).
The compounds of the formula I can be used as medicaments in the form of pharmaceutical preparations which contain them, optionally with 15 tolerable pharmaceutical vehicles. The compounds can be used as therapeutics, e.g. in the form of pharmaceutical preparations, which contain these compounds as a mixture with a pharmaceutical, organic or inorganic excipient suitable for enteral, percutaneous or parenteral administration, or administration by inhalation, such as, for example, water, CA 022~3282 1998-10-29 gum arabic, gelatin, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, petroleum jelly etc.
For this purpose, they can be administered orally in doses from 0.1 to 25 5 mg/kg/day, preferably 1 to 5 mg/kg/day, or parenterally in doses of 0.01 to 5 mg/kg/day, preferably 0.01 to 2.5 mg/kg/day, in particular 0.5 to 1.0 mg/kg/day. In severe cases, the dose can also be increased. In many cases, however, lower doses are sufficient. These details relate to an adult approximately 75 kg in weight.
3-Methoxypyridine-2-carboxylic acid is known from J. Org. Chem. 36, 2002 (1971) by E.V. Brown, M.B. Shambhu (cf. formula ll; R1-R3 = H, R4 = Me, R5 = H).
15 Furthermore, methyl 3-methoxypyridine-2-carboxylate (cf. formula ll, R5 =
CH3) is known from Acta Chem. Scand. 23, 1791 (1969).
3-Methoxypyridine-2-carboxylic acid, which was used as starting material in the examples below, was obtained starting from 4-chloro-3-methoxy-2-20 methylpyridine, which was obtained from maltol (cf. EP-A-0 208 452 and EP-A-0 304 732).
Under the examples described below, the compounds of the formula I
according to the invention are designated as substituted pyridine-2-25 carboxylic acid (glycyl ester)amides.
This manner of notation is understood as meaning substituted pyridine-2-carboxylic acid N-((alkoxycarbonyl)methyl)amides.
Classification as substituted N-(pyridyl-2-carbonyl)glycines is a further possibility.
Example 1 N-(((1 -Butyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide a) 3-Methoxypyridine-2-carboxylic acid hydrochloride a1.) 4-Chloro-3-methoxy-2-methylpyridine 1-oxide 11.2 9 (80.5 mmol) of 3-methoxy-2-methyl4(1H)-pyridone in 100 ml of phosphorus oxychloride were heated under reflux for 10 hours.
CA 022~3282 1998-10-29 The mixture was subsequently concentrated, admixed 2 ml with 30 ml of toluene each time and concentrated again, and the residue was taken up in 150 ml of water, adjusted to pH 11 using K2CO3 and extracted with dichloromethane, and the organic phase was washed with water, dried and freed of solvent. The light-brown oil (9 g) gave, using m-chloroperbenzoic acid in dichloromethane under standard conditions, 8 g of the product, m.p. 88 to 89~C (from petroleum ether).
10 a2.) 4-Chloro-2-hydroxymethyl-3-methoxypyridine 30 g (173 mmol) of 4-chloro-3-methoxy-2-methylpyridine N-oxide were dissolved in 100 ml of glacial acetic acid and admixed dropwise with stirring at 80~C with 150 ml of acetic anhydride, and the mixture was stirred at 110~C for 2 hours. The mixture was then cooled to 80~C, 200 ml of methanol were added dropwise, the mixture was heated to the boil for 15 minutes and, after cooling, concentrated under reduced pressure and the residue was taken up in methanol and poured into 300 ml of 1.5 N methanolic sodium hydroxide solution, stirred at 20~C for 30 minutes and concentrated under reduced pressure, the residue was taken up in water and extracted three times with dichloromethane, the organic phase was dried and concentrated and the residue was crystallized with petroleum ether. This gave 23 g of product, m.p. 64 to 66~C.
25 a3.) 4-Chloro-3-methoxypyridine-2-carboxylic acid At 20~C, 52.2 g (0.3 mol) of the above alcohol were added to 24 g of KOH dissolved in 900 ml of water, and the mixture was admixed a little at a time, in each case until discoloration, with 70 g (0.44 mol) of potassium permanganate at 50 to 60~C. After 1 hour of stirring at 50~C, the hot mixture was filtered off with suction, the filter cake was rinsed three times with hot water and the filtrate was concenllated under reduced pressure to 300 ml and, with cooling, adjusted to pH
1 using concentrated HCI. Filtration with suction and drying gave 50 g of product, m.p., 121~C (n. decomp.).
a4.) 29 g (0.15 mol) of the above 4-chloropyridinecarboxylic acid in 500 ml of methanol/tetrahydrofuran (1:1) were hydrogenated in a hydrogenation apparatus with Pd/C (10%). After 3.1 l of hydrogen CA 022~3282 1998-10-29 ~O 97141103 PCT/EP96/01793 had been taken up, the catalyst was removed by filtration with suction, the filtrate was concentrated and the crystalline residue was treated with ethyl acetate, filtered offwith suction and dried; yield 29 9, m.p. 170~C (with decomp.).
b) 3.8 9 (20 mmol) of 3-methoxypyridine-2-carboxylic acid hydrochloride were suspended in 500 ml of anhydrous dichloromethane and, at 20~C and with stirring, admixed with 6.1 9 (20 mmol) of glycine 1-butyl ester tosylate (prepared from glycine, 1-butanol, p-toluenesulfonic acid on a water separator with toluene), then with 7.5 ml (60 mmol) of N-ethylmorpholine, 3 9 (22.5 mmol) of 1-hydroxy-1H-benzotriazole and 8.5 9 (20 mmol) of N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide metho-p-toluenesulfonate (CMC) and stirred at 20~C for 24 hours. Undissolved particles were then filtered off with suction and the filtrate was extracted successively with aqueous Na bicarbonate solution, with 1N aqueous hydrochloric acid and with water. The organic phase was dried and concentrated under reduced pressure and the oily residue was crystallized by scratching. This gave 1.76 9 of the colorless title compound, m.p. 60 to 62~C.
Example 2 N-(((1 -Octyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide 25 2.5 9 (13 mmol) of 3-methoxypyridine-2-carboxylic acid hydrochloride, 5.5 ml (45 mmol) of N-ethylmorpholine, 2 9 (15 mmol) of 1-hydroxy-1H-benzotriazole, 4.7 9 (13 mmol) of glycine octyl ester tosylate (prepared on a water separator with toluene from glycine, 1-octanol and p-toluenesulfonic acid) and 6.3 9 (15 mmol) of CMC (cf. Example 1) were 30 stirred in 350 [lacuna] of anhydrous dichloromethane for 48 hours. After work-up by the method of Example 1), the crude product was chromatographed over silica gel using dichloromethane (with an addition of up to 2.5% of methanol in the course of the elution). This gave 3.6 9 of the colorless oily title compound, 1H NMR (CDCI3): ~ = 4.26 (d, CH2-glycine).
CA 022~3282 1998-10-29 Example 3 N-(((1 -Dodecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide By the method of Example 1, 2.8 g (15 mmol) of 3-methoxypyridine-2-5 carboxylic acid hydrochloride in 400 ml of anhydrous dichloromethane were stirred at 20~C with 6.2 9 (15 mmol) of glycine 1-dodecyl ester tosylate (prepared on a water separator from 38 g (0.5 mol) of glycine, 93.2 9 (0.5 mol) of 1-dodecanol and 115 9 (0.6 mol) of p-toluenesulfonic acid; 140 9 of product were crystallized with diethyl ether, m.p. about 106~C, sintered at 80~C), 8.2 ml (60 mmol) of N-ethylmorpholine, 2 9 (15 mmol) of 1-hydroxy-1H-benzotriazole and 6.3 9 (15 mmol) of CMC for 24 hours. Owing to a strong emulsion formation, for work-up, similarly to Example 1, dichloromethane was distilled off under reduced pressure and the residue was taken up in diethyl ether. The organic phase was dried 15 with magnesium sulfate and concentrated under reduced pressure, and 4.7 9 of the resulting yellow oil were then chromatographed over silica gel using ethyl acetate. This gave 4 9 of a light-yellow oil which crystallized after standing for 2 days after scratching with a glass rod. 3.6 9 of the colorless title compound, m.p. 47 to 49~C, were isolated.
Example 4 N-(((1 -Pentyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 5 25 N-(((1-Hexyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 6 N-(((1 -Heptyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
30 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.
CA 022~3282 1998-10-29 Example 10 N-(((1 -Octadecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 11 5 N-(((1-Propyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 12 N-(((2-Propyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
10 Example 13 N-(((1 -Tridecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
Example 14 N-(((1 -Hexadecyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide.
5.7 9 (30 mmol) of 4-chloro-3-methoxypyridine-2-carboxylic acid and 14.2 g (30 mmol) of glycine hexadecyl ester tosylate (m.p. about 90~C, prepared from glycine,1-hexadecanol, p-toluenesulfonic acid on a water separator using toluene) in 300 ml of dichloromethane were admixed by the method 20 of Example 1) with 7.7 [lacuna] (60 mmol) of N-ethylmorpholine, 4.5 g (33 mmol) of 1-hydroxy-1 H-benzotriazole and 12.8 9 (30 mmol) of N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide metho-p-toluenesulfonate (CMC) and stirred for 24 hours. Undissolved particles were then filtered off with suction and the filtrate was extracted with aqueous Na bicarbonate 25 solution, with water and with aqueous hydrochloric acid, the organic phase was concentrated and the residue (14 9) was dissolved in 500 ml of tetrahydrofuran/methanol (1:1), admixed with Pd/C (10%) and hydrogenated in a hydrogenation apparatus.
After the hydrogen uptake had ended, the catalyst was filtered off with 30 suction, the filtrate was concentrated and the residue was chromatographed over silica gel using ethyl acetate. The appropriate fractions were concentrated and the residue was crystallized using diisopropyl ether. This gave 2.1 9 of the title compound as a colorless substance, m.p. 63 to 65~C.
CA 022~32X2 1998-10-29 Example 15 N-((Ethyloxycarbonyl)methyl)-3-methoxypyridine-2-carboxamide hydrochloride 5 a) N-((Ethyloxycarbonyl)methyl)-4-chloro-3-methoxypyridine-2-carboxamide For the preparation, 4.7 9 (25 mmol) of 4-chloro-3-methoxypyridine-2-carboxylic acid were suspended in 200 ml of anhydrous dichloromethane and, at 20~C and with stirring, admixed successively with 3.5 9 (25 mmol) of glycine ethyl ester hydrochloride, 6.4 ml (50 mmol) of N-ethylmorpholine, 3.8 9 (28 mmol) of 1-hydroxy-(1 H)-benzotriazole and 5.15 9 (25 mmol) of N,N'-dicyclohexylcarbodiimide and stirred at 20~C for 20 hours.
Undissolved particles were then filtered off, the organic phase was shaken with saturated aqueous sodium carbonate solution, dried and concentrated under reduced pressure and the residue (6 9 of oil) was chromatographed over silica gel using ethyl acetate, giving 5.4 9 of an oily product.
20 B) The title compound was obtained by hydrogenating the above substance with Pd/C (10%) in a hydrogenation apparatus, m.p. 141 - to 142~C (with evolution of gas, from diethyl ether).
Example 16 25 N-(((2-Nonyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide racemate .
2.5 9 (10 mmol) of N-(carboxymethyl)-3-methoxypyridine-2-carboxamide hydrochloride (m.p. 157~C, with evolution of gas) were suspended in 30 100 ml of anhydrous tetrahydrofuran and admixed with 1.6 ml (12 mmol) of triethylamine and then dropwise with stirring with 2.4 9 of pivaloyl chloride dissolved in a little tetrahydrofuran (the temperature rises to 35-40~C).
After 30 min, the mixture was concentrated under reduced pressure and the reddish residue was taken up in 100 ml of anhydrous tetrahydrofuran 35 and admixed with 1.6 ml of triethylamine, and the mixture was then added at 20~C to a solution of Na 2-nonanolate in 2-nonanol (prepared from 30 ml of 2-nonanol and 0.8 9 (20 mmol) of NaH). After 1 h, the mixture was concentrated under reduced pressure, the residue was admixed with .. .. . ..
CA 022~32X2 1998-10-29 dichloromethane and extracted with 2N of aqueous ammonium chloride solution, the organic phase was dried and concentrated under reduced pressure and the residue (9 g) was chromatographed over silica gel using ethyl acetate. This gave 1.1 9 of the oily colorless title compound, 5 1H NMR (DMSO): ~ = 3.95 (d, CH2-glycine).
Example 17 N-(((4-Heptyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide 10 2.5 g (10 mmol) of N-(carboxymethyl)-3-methoxypyridine-2-carboxamide hydrochloride were treated as in Example 16 and then admixed at 20~C
with 140 ml of a solution of Na 4-heptanolate in 4-heptanol (prepared from 140 ml of 4-heptanol and 0.6 g (25 mmol) of sodium, ultrasonic bath). After 30 minutes, the mixture was heated at 70 to 80~C for 1 hour and, after -15 cooling, concentrated under reduced pressure, the residue was taken up inwater and extracted with dichloromethane and the organic phase was concentrated under reduced pressure and dried at an oil pump. The oily crude product crystallized after approximately 15 hours, m.p. 75 to 78~C.
20 Example 18 N-(((3-Pentyloxy)carbonyl)methyl)-3-methoxypyridine-2-carboxamide Example 19 N-(((1 -(cis-9-Octadecenyl)oxy)carbonyl)methyl)-3-methoxypyridine-2-25 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 35 Example 22 N-((Methyloxycarbonyl)methyl)-3-methoxypyridine-2-carboxamide . .
Claims (22)
1. A compound of the formula I
in which R1, R2 and R3 are hydrogen, R4 is (C1-C6)-alkyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is the radical of an alcohol GOH, including the physiologically active salts.
in which R1, R2 and R3 are hydrogen, R4 is (C1-C6)-alkyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is the radical of an alcohol GOH, including the physiologically active salts.
2. The compound of the formula I as claimed in claim 1, in which R1, R2 and R3 are hydrogen, R4 is (C1-C6)-alkyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is a branched or unbranched or cyclic (C1-C20)-alkyl radical, or a branched or unbranched or cyclic (C2-C20)-alkenyl radical, a corresponding (C2-C20)-alkinyl radical, a corresponding (C4-C20)-alkeninyl radical or a retinyl radical, where the radicals can in each case contain one or more multiple bonds, or is a phenylalkyl radical, where the above radicals in particular contain one or more substituents selected from the group consisting of hydroxyl, halogen, cyano, trifluoromethyl, carboxyl, (C1-C12)-alkyl, (C3-C8)-cycloalkyl, (C5-C8)-cycloalkenyl, (C1-C12)-alkoxy, (C1-C12)-alkoxy-(C1-C12)-alkyl, (C1-C12)-alkoxy-(C1-C12)-alkoxy, phenyl-(C1-C4)-alkyloxy, (C1-C8)-hydroxyalkyl, (C1-C12)-alkylcarbonyloxy, (C3-C8)-cycloalkylcarbonyloxy, benzoyloxy, phenyl-(C1-C4)-alkylcarbonyloxy, including the physiologically active salts.
3. The compound of the formula I as claimed in claim 1 or 2, in which R1, R2 and R3 are hydrogen, R4 is (C1-C4)-alkyl, A is a -CH2 group, in which a hydrogen atom can be replaced by a methyl group, and B is -CO2G, where G is a branched or unbranched or cyclic aliphatic (C1-C18)-alkyl radical, a (C3-C8)-cycloalkyl-(C1-C8)-alkyl radical, a branched or unbranched (C2-C18)-alkenyl radical, such as, for example, a geranyl, farnesyl or a retinyl radical, or a corresponding (C2-C18)-alkinyl radical, a benzyl, phenethyl, phenylpropyl or phenylbutyl radical, where the above radicals contain a substituent from the group consisting of hydroxyl, (C1-C4)-alkoxy, (C1-C6)-alkylcarbonyl oxy, (C3-C8)-cycloalkylcarbonyloxy, benzoyloxy or phenyl-(C1-C4)-alkylcarbonyloxy, including the physiologically active salts.
4. The compound of the formula I as claimed in claims 1 to 3, in which R1, R2 and R3 are hydrogen, R4 is (C1-C4)-alkyl, A is a -CH2 group, in which a hydrogen atom can be replaced by a methyl group, and B is -CO2G, where G is a branched or unbranched or cyclic aliphatic (C1-C18)-alkyl radical, a (C3-C8)-cycloalkyl-(C1-C4)-alkyl radical, a branched or unbranched (C2-C18)-alkenyl radical, a benzyl, phenethyl, phenylpropyl or phenylbutyl radical, including the physiologically active salts.
5. The compound of the formula I as claimed in claims 1 to 4, in which R1, R2 and R3 are hydrogen, R4 is methyl, A is a -CH2 group, in which a hydrogen can be replaced by a methyl group, and B is -CO2G, where G is a branched or unbranched (C1-C18)-alkyl or (C2-C18)-alkenyl radical, including the physiologically active salts.
6. The compound of the formula I as claimed in claims 1 to 5, in which R1, R2 and R3 are hydrogen, R4 is methyl, A is a -CH2 group and B is -CO2G, where G is a branched or unbranched (C1-C18)-alkyl or (C2-C18)-alkenyl radical, including the physiologically active salts.
7. The compound of the formula I as claimed in claims 1 to 6, in which R1, R2 and R3 are hydrogen, R4 is methyl, A is a -CH2 group, B is -CO2G, where G is a linear (C1-C18)-alkyl radical, including the physiologically active salts.
8. A process for preparing compounds of the formula I as claimed in claims 1 to 7, in which A is a -CH2 group in which a hydrogen can be replaced by a methyl group, and B is CO2G, which comprises i1.) reacting pyridine-2-carboxylic acids of the formula II (R5=H) with the amino esters [lacuna] formula III or their salts to give the amido esters of the formula I; or i2.) reacting pyridine-2-carboxylic acid esters of the formula II
(R5 = (C1-C16)-alkyl under the conditions of aminolysis to give the compounds of the formula I; or ii) esterifying the compounds of the formula IV with an alcohol GOH; or iii) alkylating the compounds of the formula V with R4X, in which X is a leaving group, in particular halogen, -OSO2Me, -OSO2phenyl, etc
(R5 = (C1-C16)-alkyl under the conditions of aminolysis to give the compounds of the formula I; or ii) esterifying the compounds of the formula IV with an alcohol GOH; or iii) alkylating the compounds of the formula V with R4X, in which X is a leaving group, in particular halogen, -OSO2Me, -OSO2phenyl, etc
9. A compound of the formula I as claimed in claims 1 to 7 for inhibiting prolyl-4-hydroxylase in vivo.
10. A compound of the formula I as claimed in claims 1 to 7 for inhibiting collagen biosynthesis.
11. A compound of the formula I as claimed in claims 1 to 7 for avoiding/reducing the deposits of connective tissue (collagens) in the diseased human organs (fibrosis).
12. A compound of the formula I as claimed in claims 1 to 7 for use in fibrotic disorders.
13. A compound of the formula I as claimed in claim 12 for use in fibrotic disorders of the lung, the liver, the kidney, the heart, the eye and the skin and in artherosclerosis.
14. A compound of the formula I as claimed in claims 12 and 13, where use takes place locally and/or systemically.
15. A compound of the formula I as claimed in claim 14 for local use for avoiding/reducing scars after surgical interventions on the human body.
16. The compound of the formula I as claimed in claims 9 to 14 for local use in the case of increased formation of connective tissue (fibrosis) in the skin.
17. The compound of the formula I as claimed in claims 9 to 14 for local use (by inhalation) in the case of increased formation of connective tissue (fibrosis) in the lung.
18. The compound of the formula I as claimed in claims 9 to 14 for local use in the case of increased formation of connective tissue (fibrosis) in the eye.
19. The compound of the formula I as claimed in claims 9 to 14 for local use in the eye for postoperative treatment of glaucoma.
20. The compound of the formula I as claimed in claims 9 to 14 for local use in radiation-induced fibrosis or fibrosis induced by chemotherapy, in particular fibroses of the lung.
21. A pharmaceutical comprising one or more compounds of the formula I as claimed in claims 1 to 8.
22. The use of the compounds for inhibiting prolyl-4-hydroxylase and in fibrotic disorders.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/EP1996/001793 WO1997041103A1 (en) | 1996-04-30 | 1996-04-30 | 3-alkoxypyridine-2-carboxylic acid amide esters, their preparation and their use as drugs |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2253282A1 true CA2253282A1 (en) | 1997-11-06 |
Family
ID=8166212
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002253282A Abandoned CA2253282A1 (en) | 1996-04-30 | 1996-04-30 | 3-alkoxypyridine-2-carboxylic acid amide esters, their preparation and their use as drugs |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP0900202A1 (en) |
| JP (1) | JP2000509047A (en) |
| KR (1) | KR20000065090A (en) |
| AU (1) | AU5762896A (en) |
| CA (1) | CA2253282A1 (en) |
| NO (1) | NO984878L (en) |
| WO (1) | WO1997041103A1 (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8323671B2 (en) | 2006-06-26 | 2012-12-04 | Akebia Therapeutics Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US8865748B2 (en) | 2011-06-06 | 2014-10-21 | Akebia Therapeutics Inc. | Compounds and compositions for stabilizing hypoxia inducible factor-2 alpha as a method for treating cancer |
| US9987262B2 (en) | 2013-11-15 | 2018-06-05 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}acetic acid, compositions, and uses thereof |
| US10150734B2 (en) | 2015-01-23 | 2018-12-11 | Akebia Therapeutics, Inc. | Solid forms of 2-(5-(3-fluorophenyl)-3-hydroxypicolinamido)acetic acid, compositions, and uses thereof |
| US10246416B2 (en) | 2011-06-06 | 2019-04-02 | Akebia Therapeutics, Inc. | Process for preparing [(3-hydroxypyridine-2-carbonyl)amino] alkanoic acids, esters and amides |
| US11324734B2 (en) | 2015-04-01 | 2022-05-10 | Akebia Therapeutics, Inc. | Compositions and methods for treating anemia |
| US11524939B2 (en) | 2019-11-13 | 2022-12-13 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino} acetic acid |
| US11713298B2 (en) | 2018-05-09 | 2023-08-01 | Akebia Therapeutics, Inc. | Process for preparing 2-[[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino]acetic acid |
| US11857543B2 (en) | 2013-06-13 | 2024-01-02 | Akebia Therapeutics, Inc. | Compositions and methods for treating anemia |
Families Citing this family (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2803592A1 (en) * | 2000-01-06 | 2001-07-13 | Aventis Cropscience Sa | NOVEL DERIVATIVES OF 3-HYDROXYPICOLINIC ACID, PROCESS FOR THEIR PREPARATION AND FUNGICIDAL COMPOSITIONS CONTAINING SAME |
| AU2002241154A1 (en) | 2001-03-21 | 2002-10-03 | Isis Innovation Ltd. | Assays, methods and means relating to hypoxia inducible factor (hif) hydroxylase |
| EP3520784A1 (en) * | 2001-12-06 | 2019-08-07 | Fibrogen, Inc. | Hif prolyl hydroxylase inhibitor for treatment of anemia |
| US7588924B2 (en) | 2006-03-07 | 2009-09-15 | Procter & Gamble Company | Crystal of hypoxia inducible factor 1 alpha prolyl hydroxylase |
| HUE026109T2 (en) | 2009-11-06 | 2016-05-30 | Aerpio Therapeutics Inc | Compositions and methods for treating colitis |
| AU2013370491B2 (en) | 2012-12-28 | 2016-10-06 | Corteva Agriscience Llc | Synergistic fungicidal mixtures for fungal control in cereals |
| KR20170099930A (en) | 2014-12-30 | 2017-09-01 | 다우 아그로사이언시즈 엘엘씨 | Use of picolinamide compounds with fungicidal activity |
| CN113615696A (en) | 2014-12-30 | 2021-11-09 | 美国陶氏益农公司 | Pyridine amide compounds having fungicidal activity |
| RU2017123807A (en) * | 2014-12-30 | 2019-01-31 | ДАУ АГРОСАЙЕНСИЗ ЭлЭлСи | PICOLINAMIDES WITH FUNGICIDAL ACTIVITY |
| BR112017013767A2 (en) | 2014-12-30 | 2018-03-13 | Dow Agrosciences Llc | use of picolinamides as fungicides |
| CA2972408A1 (en) | 2014-12-30 | 2016-07-07 | Dow Agrosciences Llc | Use of picolinamide compounds with fungicidal activity |
| US20180243271A1 (en) * | 2015-05-28 | 2018-08-30 | Japan Tobacco Inc. | Method for Treating or Preventing Diabetic Nephropathy |
| KR102659552B1 (en) | 2017-05-02 | 2024-04-23 | 코르테바 애그리사이언스 엘엘씨 | Use of acyclic picolinamide compounds as fungicides against fungal diseases on turf grasses |
| TWI774761B (en) | 2017-05-02 | 2022-08-21 | 美商科迪華農業科技有限責任公司 | Synergistic mixtures for fungal control in cereals |
| TW201842851A (en) | 2017-05-02 | 2018-12-16 | 美商陶氏農業科學公司 | Synergistic mixtures for fungal control in cereals |
| BR102019004480B1 (en) | 2018-03-08 | 2023-03-28 | Dow Agrosciences Llc | PICOLINAMIDES AS FUNGICIDES |
| WO2020081382A1 (en) | 2018-10-15 | 2020-04-23 | Dow Agrosciences Llc | Methods for sythesis of oxypicolinamides |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE59401923D1 (en) * | 1993-11-02 | 1997-04-10 | Hoechst Ag | Substituted heterocyclic carboxylic acid amide esters, their preparation and their use as medicaments |
-
1996
- 1996-04-30 CA CA002253282A patent/CA2253282A1/en not_active Abandoned
- 1996-04-30 AU AU57628/96A patent/AU5762896A/en not_active Abandoned
- 1996-04-30 KR KR1019980708679A patent/KR20000065090A/en not_active Application Discontinuation
- 1996-04-30 WO PCT/EP1996/001793 patent/WO1997041103A1/en not_active Application Discontinuation
- 1996-04-30 JP JP09538479A patent/JP2000509047A/en active Pending
- 1996-04-30 EP EP96914152A patent/EP0900202A1/en not_active Withdrawn
-
1998
- 1998-10-19 NO NO984878A patent/NO984878L/en not_active Application Discontinuation
Cited By (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11426393B2 (en) | 2006-06-26 | 2022-08-30 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and methods of use |
| USRE47437E1 (en) | 2006-06-26 | 2019-06-18 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US11883386B2 (en) | 2006-06-26 | 2024-01-30 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US8722895B2 (en) | 2006-06-26 | 2014-05-13 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and method of use |
| US10729681B2 (en) | 2006-06-26 | 2020-08-04 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US8940773B2 (en) | 2006-06-26 | 2015-01-27 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US9598370B2 (en) | 2006-06-26 | 2017-03-21 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US8323671B2 (en) | 2006-06-26 | 2012-12-04 | Akebia Therapeutics Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US8343952B2 (en) | 2006-06-26 | 2013-01-01 | Akebia Therapeutics Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US8598210B2 (en) | 2006-06-26 | 2013-12-03 | Akebia Therapeutics, Inc. | Prolyl hydroxylase inhibitors and methods of use |
| US8865748B2 (en) | 2011-06-06 | 2014-10-21 | Akebia Therapeutics Inc. | Compounds and compositions for stabilizing hypoxia inducible factor-2 alpha as a method for treating cancer |
| US11267785B2 (en) | 2011-06-06 | 2022-03-08 | Akebia Therapeutics, Inc. | Process for preparing [(3-hydroxypyridine-2-carbonyl)amino]alkanoic acids, esters and amides |
| US10246416B2 (en) | 2011-06-06 | 2019-04-02 | Akebia Therapeutics, Inc. | Process for preparing [(3-hydroxypyridine-2-carbonyl)amino] alkanoic acids, esters and amides |
| US10738010B2 (en) | 2011-06-06 | 2020-08-11 | Akebia Therapeutics, Inc. | Process for preparing [(3-hydroxypyridine-2-carbonyl)amino] alkanoic acids, esters and amides |
| US11857543B2 (en) | 2013-06-13 | 2024-01-02 | Akebia Therapeutics, Inc. | Compositions and methods for treating anemia |
| US11690836B2 (en) | 2013-11-15 | 2023-07-04 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}acetic acid, compositions, and uses thereof |
| US11065237B2 (en) | 2013-11-15 | 2021-07-20 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}acetic acid, compositions, and uses thereof |
| US9987262B2 (en) | 2013-11-15 | 2018-06-05 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}acetic acid, compositions, and uses thereof |
| US10149842B2 (en) | 2013-11-15 | 2018-12-11 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}acetic acid, compositions, and uses thereof |
| US10596158B2 (en) | 2013-11-15 | 2020-03-24 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino}acetic acid, compositions, and uses thereof |
| US10150734B2 (en) | 2015-01-23 | 2018-12-11 | Akebia Therapeutics, Inc. | Solid forms of 2-(5-(3-fluorophenyl)-3-hydroxypicolinamido)acetic acid, compositions, and uses thereof |
| US11324734B2 (en) | 2015-04-01 | 2022-05-10 | Akebia Therapeutics, Inc. | Compositions and methods for treating anemia |
| US11844756B2 (en) | 2015-04-01 | 2023-12-19 | Akebia Therapeutics, Inc. | Compositions and methods for treating anemia |
| US11713298B2 (en) | 2018-05-09 | 2023-08-01 | Akebia Therapeutics, Inc. | Process for preparing 2-[[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino]acetic acid |
| US11524939B2 (en) | 2019-11-13 | 2022-12-13 | Akebia Therapeutics, Inc. | Solid forms of {[5-(3-chlorophenyl)-3-hydroxypyridine-2-carbonyl]amino} acetic acid |
Also Published As
| Publication number | Publication date |
|---|---|
| WO1997041103A1 (en) | 1997-11-06 |
| KR20000065090A (en) | 2000-11-06 |
| NO984878L (en) | 1998-12-29 |
| EP0900202A1 (en) | 1999-03-10 |
| AU5762896A (en) | 1997-11-19 |
| JP2000509047A (en) | 2000-07-18 |
| NO984878D0 (en) | 1998-10-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2253282A1 (en) | 3-alkoxypyridine-2-carboxylic acid amide esters, their preparation and their use as drugs | |
| AU743750B2 (en) | 3-hydroxypyridine-2-carboxamidoesters, their preparation and their use as pharmaceuticals | |
| US6093730A (en) | Substituted isoquinoline-3-carboxamides, their preparation and their use as pharmaceuticals | |
| EP0661269B1 (en) | Substituted heterocyclic carboxylic acid amides, their preparation and their use as medicaments | |
| US5658933A (en) | Substituted heterocyclic carboxamide esters, their preparation and their use as pharmaceuticals | |
| RU2129545C1 (en) | Sulfamido- and sulfamido-carbonylpyridine-2-carboxylic acid amides, method of their synthesis and pharmaceutical composition | |
| TW399046B (en) | Substituted heterocyclic carboxamides, their preparation and their use as pharmaceuticals | |
| US4968670A (en) | Pyridine-2,4- and 2,5-dicarboxylic acid derivatives, a process for their preparation, the use thereof, and medicaments based on these compounds | |
| IE880350L (en) | Pyridine-2,4- and -2,5-dicarboxylic acid derivatives,¹process for their preparation, the use thereof and¹medicaments based on these compounds | |
| RU2117660C1 (en) | Esters of acylsulfonamido- and sulfonamidopyridine-2-carboxylic acid, pharmaceutical composition containing thereof and a method of its preparing | |
| MXPA97009438A (en) | Ester-amidas de acido 3-hidroxi-piridina-2-carboxilico, its preparation and its use as a medicamen | |
| RU1836349C (en) | Substituted diamides of 2,4- or 2,5-pyridinedicarboxylic acid possessing lysin- or prolinehydroxylase inhibiting activity | |
| CA2063177A1 (en) | Mixed pyridine-2,4- and -2,5-dicarboxamides, a process for preparing them, the use thereof and pharmaceuticals based on these compounds |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Dead |