HK1196603B - 2-thiopyrimidinones - Google Patents
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- HK1196603B HK1196603B HK14109952.1A HK14109952A HK1196603B HK 1196603 B HK1196603 B HK 1196603B HK 14109952 A HK14109952 A HK 14109952A HK 1196603 B HK1196603 B HK 1196603B
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Description
Background
The present invention relates to compounds which are inhibitors of myeloperoxidase, pharmaceutical compositions comprising such inhibitors and the use of such inhibitors for the treatment of, for example, cardiovascular conditions including acute coronary syndrome.
Myeloperoxidase (MPO) is a heme-containing enzyme belonging to the peroxidase superfamily. Examples of animal peroxidases are lactoperoxidase, thyroid peroxidase, eosinophil peroxidase and myeloperoxidase. Myeloperoxidase is present in the primary granules of neutrophils and to a lesser extent in monocytes. Which catalyzes the synthesis of hypochlorous acid from chloride and hydrogen peroxide. The hypochlorous acid formed is a powerful oxidizing agent that can react with a variety of cellular substrates including heme proteins, porphyrins, thiols, iron-sulfur centers, nucleotides, DNA, unsaturated lipids, amines, and amino acids.
In addition, MPO-catalyzed reactions and their products have been found to exhibit atherogenic biological activity during the development of atherosclerosis and cardiovascular disease. For example, myeloperoxidase plasma levels are associated with the appearance of cardiovascular disease in patients suffering from unstable angina. Myeloperoxidase has been reported to contribute to the development of atherosclerosis by oxidation of lipids and proteins in LDL and HDL.
Furthermore, MPO-producing oxidants have been observed to reduce the bioavailability of nitric oxide, an important vasodilator. Thus, high MPO plasma concentrations are inversely correlated with the success of establishing reperfusion therapy of occluded blood vessels. High MPO concentrations are also associated with decreased survival from congestive heart failure. Furthermore, MPO has been shown to play a role in plaque instability (which leads to plaque rupture and myocardial infarction).
Thus, MPO is considered to play a role in several cardiovascular disease-causing processes, including 1) reduction of cholesterol transport and progression of atherosclerotic plaques toward unstable states, 2) instability and plaque rupture of atherosclerotic plaques, 3) consumption of nitric oxide leading to impaired endothelial function and flow, and 4) pathological tissue damage following ischemia contributing to atrial fibrillation and adverse cardiac remodeling with left ventricular hypertrophy leading to congestive heart failure. Inhibitors of MPO activity have therefore been proposed to offer significant medical advantages in the prevention and treatment of cardiovascular disease.
Although MPO has been widely implicated in the etiology and progression of cardiovascular disease, biologically safe and non-toxic inhibitors of MPO have yet to be developed. Thus, there remains a need for agents that have myeloperoxidase inhibitory activity and that can be used to treat, prevent or reduce the manifestations of the diseases described herein.
Summary of The Invention
The invention relates to compounds of formula I
Or a pharmaceutically acceptable salt or prodrug thereof, wherein
R1Is a five to six membered aromatic ring optionally having one to three heteroatoms independently selected from nitrogen, sulfur and oxygen or a bicyclic ring consisting of two fused partially saturated, fully saturated or fully unsaturated five to six membered rings independently optionally having one to four heteroatoms independently selected from nitrogen, sulfur and oxygen; and is
The R is1Optionally, independently by cyano, halogen, hydroxy, amino, (C)1-C4) Alkyl, (C)1-C4) Alkoxy group, (C)1-C4) Alkoxy (C)1-C4) Alkyl, hydroxy (C)2-C4) Alkoxy, carbamoyl (C)1-C4) Alkoxy, amino (C)2-C4) Alkoxy, cyano (C)1-C4) Alkyl, mono-N-or di-N, N- (C)1-C4) Alkylamino, aminocarbonyl, mono-N-or di-N, N (C)1-C4) An alkylaminocarbonyl group,(C1-C4) Alkylthio, aminosulfonyl, (C)1-C4) Alkylsulfinyl (C)1-C4) Alkylsulfonyl radicals or mono-N-or di-N, N (C)1-C4) Alkylaminosulfonyl mono-, di-or tri-substituted, wherein (C)1-C4) Alkyl or (C)1-C4) Any of the alkoxy groups may be optionally mono-, di-, or tri-substituted with fluorine; or wherein R is1Optionally substituted with a five to six membered aromatic ring optionally having one to three heteroatoms independently selected from nitrogen, sulfur, and oxygen;
R2is a fully saturated, partially unsaturated, or fully unsaturated, one to fourteen membered straight carbon chain, wherein the carbons, other than the linking carbon,
a. it may be a branched chain or a branched chain,
b. optionally substituted with one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, wherein said sulfur is optionally mono-or di-substituted with oxo,
c. optionally, independently mono-, di-or tri-substituted with halogen,
d. optionally mono-substituted by hydroxy, and
e. optionally mono-substituted by oxo,
and wherein the carbon chain is optionally mono-substituted with Z;
wherein Z is a partially saturated, fully saturated or fully unsaturated three-to seven-membered ring optionally having one to three heteroatoms independently selected from oxygen, sulfur and nitrogen, or a bicyclic ring consisting of two fused partially saturated, fully saturated or fully unsaturated five-to six-membered rings independently optionally having one to four heteroatoms independently selected from nitrogen, sulfur and oxygen;
wherein said Z is optionally, independently, substituted by halogen, (C)1-C6) Alkyl, (C)1-C6) Alkylcarbonyl, aminothio, amino (C)1-C6) An alkylcarbonyl group,Hydroxy, diaminomethylene, carbamoyl or (C)1-C6) Alkoxy is mono-, di-or tri-substituted, and wherein (C) is1-C6) Alkyl or (C)1-C6) Alkoxy substituents are further optionally substituted with one to three halogens, and wherein (C) is1-C6) Alkyl or (C)2-C6) The alkoxy substituents are further optionally substituted with one to three hydroxy groups;
provided that R is1Is not phenyl, and R2Is not (C)1-C6) An alkyl group.
Yet another aspect of this invention is directed to a method of treating a cardiovascular condition (condition) in a mammal (including a human, male or female) by administering to a mammal in need of such treatment a therapeutically effective amount of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug.
Also provided herein are compositions comprising a pharmaceutically effective amount of one or more compounds described herein and a pharmaceutically acceptable carrier, excipient (vehicle) or diluent.
The present invention also relates to a pharmaceutical combination composition (combination composition) comprising: a therapeutically effective amount of a composition comprising:
a first compound which is a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug;
a second compound that is an angiotensin converting enzyme inhibitor, an HMG-CoA reductase inhibitor, a non-steroidal anti-inflammatory drug, a factor Xa inhibitor, or warfarin; and/or optionally
A pharmaceutically acceptable carrier, excipient (vehicle) or diluent.
The present invention relates to compounds of formula IA,
or a pharmaceutically acceptable salt or prodrug thereof
Wherein
R1Is a five to six membered aromatic ring optionally having one to three heteroatoms independently selected from nitrogen, sulfur and oxygen or a bicyclic ring consisting of two fused partially saturated, fully saturated or fully unsaturated five to six membered rings independently optionally having one to four heteroatoms independently selected from nitrogen, sulfur and oxygen; and is
The R is1Optionally, independently by cyano, halogen, hydroxy, amino, (C)1-C4) Alkyl, (C)1-C4) Alkoxy group, (C)1-C4) Alkoxy (C)1-C4) Alkyl, hydroxy (C)2-C4) Alkoxy, carbamoyl (C)1-C4) Alkoxy, amino (C)2-C4) Alkoxy, cyano (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonyloxy (C)1-C4) Alkyl, amino (C)1-C4) Alkylcarbonyloxy (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonyloxy (C)1-C4) Alkoxy, amino (C)1-C4) Alkylcarbonyloxy (C)1-C4) Alkoxy, mono-N-or di-N, N- (C)1-C4) Alkylamino, aminocarbonyl, mono-N-or di-N, N (C)1-C4) Alkylaminocarbonyl, (C)1-C4) Alkylthio, aminosulfonyl, (C)1-C4) Alkylsulfinyl (C)1-C4) Alkylsulfonyl radicals or mono-N-or di-N, N (C)1-C4) Alkylaminosulfonyl mono-, di-or tri-substituted, wherein (C)1-C4) Alkyl or (C)1-C4) Any of the alkoxy groups may be optionally mono-, di-, or tri-substituted with fluorine; or wherein R is1Optionally is optionally provided with one toFive to six membered aromatic ring substitution of three heteroatoms independently selected from nitrogen, sulfur and oxygen;
R2is a fully saturated, partially unsaturated, or fully unsaturated, one to fourteen membered straight carbon chain, wherein the carbons, other than the linking carbon,
a. it may be a branched chain or a branched chain,
b. optionally substituted with one or two heteroatoms independently selected from oxygen and sulfur, and optionally substituted with one to four nitrogens, wherein said sulfur is optionally mono-or di-substituted with oxo,
c. optionally, independently mono-, di-or tri-substituted with halogen,
d. optionally mono-substituted by hydroxy, and
e. optionally mono-substituted by oxo,
and wherein the carbon chain is optionally mono-substituted with Z;
wherein Z is a partially saturated, fully saturated or fully unsaturated three-to seven-membered ring optionally having one to three heteroatoms independently selected from oxygen, sulfur and nitrogen, or a bicyclic ring consisting of two fused partially saturated, fully saturated or fully unsaturated five-to six-membered rings independently optionally having one to four heteroatoms independently selected from nitrogen, sulfur and oxygen;
wherein said Z is optionally, independently, substituted by amino, halogen, (C)1-C6) Alkyl, (C)1-C6) Alkylcarbonyl, aminothio, amino (C)1-C6) Alkylcarbonyl, hydroxy, diaminomethylene, carbamoyl or (C)1-C6) Alkoxy is mono-, di-or tri-substituted, and wherein (C) is1-C6) Alkyl or (C)1-C6) Alkoxy substituents are further optionally substituted with one to three halogens, and wherein (C) is1-C6) Alkyl or (C)2-C6) The alkoxy substituents are further optionally substituted with one to three hydroxy groups;
provided that R is1Is not unsubstituted phenyl, and R2Not unsubstituted (C)1-C6) An alkyl group.
The present invention also relates to methods of treating cardiovascular events and conditions comprising administering to a mammal in need of such treatment a therapeutically effective amount of a compound of formula IA or a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug, wherein the cardiovascular condition or event is heart failure, congestive heart failure, peripheral artery disease, pulmonary hypertension, vasculitis, primary or secondary myocardial infarction, ischemia reperfusion injury, atrial fibrillation or coronary artery bypass graft surgery (CABG).
The present invention also relates to methods of treating a condition comprising administering to a mammal in need of such treatment a therapeutically effective amount of a compound of formula IA or a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug, wherein the condition is diabetes, renal insufficiency, dialysis, delayed recovery of graft function, transplant organ rejection, or nephropathy caused by a contrast agent.
Also provided herein are compositions comprising a pharmaceutically effective amount of one or more compounds of formula IA as described herein, and a pharmaceutically acceptable carrier, excipient (vehicle) or diluent.
The present invention also relates to a pharmaceutical combination composition comprising: a therapeutically effective amount of a composition comprising:
a first compound which is a compound of formula IA, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or of said prodrug;
a second compound that is an angiotensin converting enzyme inhibitor, an HMG-CoA reductase inhibitor, a non-steroidal anti-inflammatory drug, a factor Xa inhibitor, or warfarin; and/or optionally
A pharmaceutically acceptable carrier, excipient (vehicle) or diluent.
All patents and patent applications mentioned herein are incorporated herein by reference.
Additional features and advantages of the invention will be set forth in the description which follows, and in the claims which follow, which illustrate the invention.
Brief Description of Drawings
FIG. 1 is a characteristic X-ray powder diffraction pattern (vertical axis: intensity (CPS); horizontal axis: 2 θ (degrees)) showing the crystalline form of example 1.
FIG. 2 is a characteristic X-ray powder diffraction pattern (vertical axis: intensity (CPS); horizontal axis: 2 θ (degrees)) showing the crystalline form of example 2.
Detailed Description
A preferred group of compounds (designated as group A) contains those compounds having the formula I shown above and wherein R is1Is phenyl, naphthyl, furyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, quinolyl, isoquinolyl, pyrazolyl, imidazolinyl, cyclopentyl, cyclohexyl, pyrrolyl, indolyl, benzo [ b ]]Thienyl, benzothiazolyl, benzo [ b ]]Furyl or thienyl; and wherein said R1Independently by cyano group, (C)1-C4) Alkyl, (C)1-C4) Alkoxy, hydroxy (C)2-C4) Alkoxy, trifluoro (C)1-C4) Alkyl, trifluoro (C)1-C4) Alkoxy or halogen mono-, di-or tri-substituted.
A preferred group of compounds from group A is designated group B, which contains compounds wherein R is2Is a fully saturated, partially unsaturated, or fully unsaturated, one to fourteen membered straight carbon chain, wherein the carbons, other than the linking carbon,
a. it may be a branched chain or a branched chain,
b. optionally substituted with one or two heteroatoms independently selected from oxygen, sulfur and nitrogen, wherein said sulfur is optionally mono-or di-substituted with oxo,
c. optionally, independently mono-, di-or tri-substituted with halogen,
d. optionally mono-substituted by hydroxy, and
e. optionally mono-substituted with oxo; or
R2Is furyl (C)1-C4) Alkyl, triazolyl (C)1-C4) Alkyl, pyridyl (C)1-C4) Alkyl, pyrazinyl (C)1-C4) Alkyl, pyridazinyl (C)1-C4) Alkyl, pyrimidinyl (C)1-C4) Alkyl, imidazolyl (C)1-C4) Alkyl or pyrrolidinyl (C)1-C4) Alkyl radical, said R2The ring is optionally substituted with (C)1-C4) Alkyl, (C)1-C4) Alkoxy or halogen mono-, di-or tri-substituted.
One preferred group of compounds from group B of compounds is designated group C, which contains compounds wherein R is1Is phenyl, naphthyl, pyridyl, quinolyl, isoquinolyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, imidazolinyl, furyl, cyclopentyl, cyclohexyl, pyrrolyl, indolyl, benzo [ b ]]Thienyl, benzothiazolyl, benzo [ b ]]Furyl or thienyl; wherein said R1Independently quilt (C)1-C4) Alkyl, (C)1-C4) Alkoxy, hydroxy (C)2-C4) Alkoxy, cyano, trifluoromethyl, trifluoromethoxy or halo mono-, di-or tri-substituted; and is
R2Is (C)1-C4) Alkoxy (C)1-C4) Alkyl, carboxyl (C)1-C4) Alkyl, mono-or di-hydroxy (C)2-C6) Alkyl, amino (C)2-C4) Alkyl, diamino methylene amino (C)2-C4) Alkyl, mono-N-or di-N, N (C)1-C4) Alkylamino radical (C)2-C4) Alkyl, (C)1-C4) Alkylcarbonyloxy (C)1-C4) Alkyl, (C)1-C4) Alkoxycarbonyl (C)1-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, carbamoylamino (C)2-C4) Alkyl, mono-N-or di-N, N (C)1-C4) Alkylcarbamoyl (C)1-C4) Alkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, (C)1-C4) Alkoxycarbonylamino (C)2-C4) Alkyl, (C)1-C4) Alkylsulfonylamino (C)2-C4) Alkyl, (C)1-C4) Alkylaminosulfonyl (C)1-C4) Alkyl, aminosulfonyl (C)1-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl or (C)1-C4) Alkylthio alkyl (C)1-C4)。
One preferred group of compounds from group C, designated as group D, contains compounds wherein R is1Is phenyl and said R1Independently mono-, di-or tri-substituted with hydroxyethoxy, methyl, methoxy, fluoro or chloro; and is
R2Is diaminomethyleneamino (C)2-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, hydroxy (C)2-C4) Alkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl or amino (C)2-C4) An alkyl group.
One preferred group of compounds from group B of compounds is designated group E, which contains compounds wherein R is1Is phenyl, naphthyl, pyridyl, quinolyl, isoquinolyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, imidazolinyl, furyl, cyclopentyl, cyclohexyl, pyrrolyl, indolyl, benzo [ b ]]Thienyl, benzothiazolyl, benzo [ b ]]Furyl or thienyl; wherein said R1Independently quilt (C)1-C4) Alkyl, (C)1-C4) Alkoxy, hydroxy (C)2-C4) Alkoxy, cyano, trifluoromethyl, trifluoromethoxy or halo mono-, di-or tri-substituted; and is
R2Is triazolyl (C)1-C4) Alkyl, pyridyl (C)1-C4) Alkyl, pyrazinyl (C)1-C4) Alkyl, pyridazinyl (C)1-C4) Alkyl, pyrimidinyl (C)1-C4) Alkyl, imidazolyl (C)1-C4) Alkyl or pyrrolidinyl (C)1-C4) Alkyl radical, said R2The ring is optionally substituted with (C)1-C4) Alkyl, (C)1-C4) Alkoxy or halogen mono-, di-or tri-substituted.
A preferred group of compounds designated as group F comprises compounds having the formula I above wherein R is1Is phenyl and said R1Independently mono-, di-, tri-substituted with hydroxyethoxy, methyl, methoxy, fluoro or chloro.
A preferred group of compounds designated group G comprises compounds having the formula I above wherein R is2Is hydroxy (C)2-C4) Alkyl, diamino methylene amino (C)2-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl or amino (C)2-C4) An alkyl group.
A preferred group of compounds designated as group H comprises compounds having the formula I above wherein R2Independently by amino, carbamoyl, hydroxy, (C)1-C4) Alkoxy, amino (C)1-C4) Alkylcarbonylamino, amino (C)2-C4) Alkylcarbamoyl, (C)1-C4) Mono-or di-substituted (C) by alkylcarbonylamino or diaminomethyleneamino1-C4) An alkyl group.
A preferred group of compounds designated as group I comprises compounds wherein the compound is
6- (2, 4-dimethoxyphenyl) -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
1- (2-aminoethyl) -6- (2, 4-dimethoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
2- [6- (2, 5-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
2- [6- (5-chloro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
1- (2-aminoethyl) -2-thioxo-6- (2,4, 5-trimethoxyphenyl) -2, 3-dihydropyrimidin-4 (1H) -one;
1- (3-aminopropyl) -6- (2-methoxy-5-methylphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
n- {2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] ethyl } glycinamide;
2- {3- [6- (2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] propyl } guanidine;
1- [ (2S) -3-amino-2-hydroxypropyl ] -6- (5-chloro-2-methoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
1- [ (2R) -3-amino-2-hydroxypropyl ] -6- (5-chloro-2-methoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] N- (2-aminoethyl) -acetamide; or
1- (2-aminoethyl) -6- [2- (2-hydroxyethoxy) phenyl ] -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one
Or a pharmaceutically acceptable salt thereof.
Particularly preferred compounds are
2- (6- (2, 5-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide, or a pharmaceutically acceptable salt thereof.
Particularly preferably, the compound is
Another particularly preferred compound is
2- (6- (5-chloro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide, or a pharmaceutically acceptable salt thereof.
Particularly preferred compounds are
One preferred group of compounds from group C, designated as group J, contains compounds wherein R is1Is naphthyl, quinolyl, isoquinolyl, indolyl, benzo [ b ]]Thienyl, benzothiazolyl, benzo [ b ]]Furyl or thienyl and said R1Independently mono-, di-or tri-substituted by hydroxyethoxy, methyl, methoxy, fluoro or chloro(ii) a And is
R2Is diaminomethyleneamino (C)2-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, hydroxy (C)2-C4) Alkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl or amino (C)2-C4) An alkyl group.
A preferred group of compounds designated as group K comprises compounds wherein the compound is
2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
2- [6- (2-methoxy-5-methylphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
1- [ (2R) -2-aminopropyl ] -6- (2, 4-dimethoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
2- [6- (3-methoxy-2-naphthyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide; or
2- [6- (1H-indol-4-yl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide
Or a pharmaceutically acceptable salt thereof.
A preferred group of compounds designated as group L comprises compounds wherein the compound is
2- {6- [2- (2-hydroxyethoxy) -5-methoxyphenyl ] -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl } acetamide;
2- {6- [2- (2-hydroxyethoxy) -4-methoxyphenyl ] -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
6- [2- (2-hydroxyethoxy) -4-methoxyphenyl ] -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
6- [ 5-fluoro-2- (2-hydroxyethoxy) phenyl ] -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one; or
2- {6- [2- (2-hydroxyethoxy) -4-methoxyphenyl ] -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl } acetamide
Or a pharmaceutically acceptable salt thereof.
A particularly preferred compound is N- (2-aminoethyl) -2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide or a pharmaceutically acceptable salt thereof.
Particularly preferably, the compound is
Preferred cardiovascular conditions include heart failure, congestive heart failure, peripheral arterial disease, pulmonary hypertension or vasculitis.
Other preferred cardiovascular conditions include unstable angina or patients who have experienced myocardial infarction.
Pharmaceutically acceptable salts of the compounds of formula I or IA include acid addition and base salts thereof. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate/carbonate, bisulfate/sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, salicylate (hibenzate), hydrochloride/chloride, hydrobromide/bromide, hydroiodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthalenedicarboxylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/biphosphate/dihydrogen phosphate, dihydrogenphosphate, dihydrogen, Pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinofoate.
Suitable base salts are formed from bases which form non-toxic salts. Examples include aluminum, arginine, calcium, choline, diethylamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, trimethylamine, and zinc salts. Hemisalts of acids and bases may also be formed, such as hemisulfate and hemicalcium salts. For a review of the appropriate Salts, see Stahl and Wermuth, Handbook of pharmaceutical Salts: properties, Selection, and Use (Wiley-VCH, 2002).
The compounds of the present invention may exist in both unsolvated and solvated forms. The term "solvate" as used herein is intended to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules, such as ethanol. The solvent molecules are known in the pharmaceutical art to be harmless to the recipient, such as water, ethanol, and the like. Other solvents may be used as intermediate solvates in the preparation of more desirable solvates such as methanol, methyl tert-butyl ether, ethyl acetate, methyl acetate, (S) -propylene glycol, (R) -propylene glycol, 1, 4-butyne-diol, and the like. When the solvent is water, the term "hydrate" is used. Pharmaceutically acceptable solvates include hydrates and other solvates in which the solvent of crystallization may be replaced by an isotope, e.g. D2O、d6-acetone, d6-dimethyl sulfoxide. The term "hydrate" means a complex in which the solvent molecule is water. The solvates and/or hydrates preferably exist in crystalline form.
Included within the scope of the invention are complexes such as clathrates, drug-host inclusion complexes, wherein (unlike the solvates described above) the drug and host are present in stoichiometric or non-stoichiometric amounts. Also included are complexes of drugs containing two or more organic and/or inorganic components, which may be in stoichiometric or non-stoichiometric amounts. The resulting complex may be ionized, partially ionized, or non-ionized. For an overview of these complexes, see J Pharm Sci, 64(8), 1269-1288 (8/1975) by Haleblian.
The compounds of the invention include compounds of formula I or IA as defined above, polymorphs, isomers (including optical, geometric and tautomeric isomers) thereof as defined below and isotopically labeled compounds of formula I or IA.
The compounds of the present invention may be administered in prodrug form. Certain derivatives of a compound of formula I or IA which themselves have little or no pharmacological activity may be converted to a compound of formula I or IA having the desired activity when administered in or on the body, for example by hydrolytic cleavage. These derivatives are referred to as "prodrugs". [ further information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems", Vol.14, ACS Symposium Series (T Higuchi and WStella) and "Bioreversible Carriers in Drug Delivery", Pergamon Press, 1987 (ed.E. BRoche, American Pharmaceutical Association) ].
Prodrugs can be prepared, for example, by substituting certain moieties known to those skilled in the art as "pro-moieties" for appropriate functional groups present in compounds of formula I or IA, as described in h.
Some examples of such prodrugs include:
(i) when a compound of formula I or IA contains a carboxylic acid function (-COOH), an ester thereof, e.g., hydrogen, with (C)1-C8) Alkyl substitution;
(ii) when the compounds of formula I or IA contain an alcohol function (-OH), they are ethers thereof, e.g. by reacting hydrogen with (C)1-C6) Alkanoyloxymethyl substitution; and
(iii) when the compounds of formula I or IA contain primary or secondary amino functions (-NH)2or-NHR, where R.noteq.H), is an amide thereof, e.g. by reacting one or two hydrogens with (C)1-C10) And (4) alkanoyl substitution.
Furthermore, certain compounds of formula I or IA may themselves act as prodrugs of other compounds of formula I or IA.
Compounds of formula I or IA containing asymmetric carbon atoms may exist as two or more stereoisomers. When a compound of formula I or IA contains an alkenyl or alkenylene group or a cycloalkyl group, geometric cis/trans (or Z/E) isomers may be present. Tautomers may occur when a compound contains, for example, a keto or oxime group or an aromatic moiety. It follows that a single compound may exhibit more than one isomeric form. For example, the following are illustrative of tautomers of compounds of formula I or IA.
Thiouracil tautomers
The most predominant tautomer
Examples of other tautomers within the scope of the claimed compounds are guanidine tautomers of the compounds illustrated below.
Examples of guanidine tautomers and geometric isomers
Included within the scope of the claimed compounds of the present invention are all stereoisomers, geometric isomers and tautomeric forms of the compounds of formula (I), including compounds exhibiting more than one isomeric form, and mixtures of one or more thereof. Also included are acid addition or base salts in which the counterion is optically active, such as D-lactate or L-lysine; or racemic, such as DL-tartrate or DL-arginine.
The present invention includes all pharmaceutically acceptable isotopically-labelled compounds of formula (I) wherein one or more atoms are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number typically found in nature.
Examples of isotopes suitable for inclusion in compounds of the invention include isotopes of hydrogen, such as2H and3h; isotopes of carbon, such as11C、13C and14c; isotopes of chlorine, such as36Cl; isotopes of fluorine, such as18F; isotopes of iodine, such as123I and125i; isotopes of nitrogen, such as13N and15n; isotopes of oxygen, such as15O、17O and18o; isotopes of phosphorus, such as32P; and isotopes of sulfur, such as35S。
Certain isotopically-labeled compounds of formula (I), for example those into which radioactive isotopes are incorporated, are useful in drug and/or substrate tissue distribution studies. Radioactive tritium (i.e., tritium)3H) And carbon-14 (i.e.14C) Are particularly useful for this purpose because of their ease of introduction and the ease of detection methods.
With heavier isotopes such as deuterium (i.e. deuterium)2H) Substitution may then result in certain medical advantages due to greater metabolic stability, e.g., increased half-life in vivo or reduced dosage requirements, and may therefore be preferred in some circumstances.
With positron emitting isotopes such as11C、18F、15O and13n substitution can then be used in Positron Emission Tomography (PET) studies to examine the occupancy of substrate receptors.
Isotopically-labelled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples and preparations using an appropriate isotopically-labelled reagent in place of the unlabelled reagent previously used. Reference herein to "treatment" or the like includes curative, palliative or prophylactic treatment.
As used herein, "reaction-inert solvent" and "inert solvent" means a solvent or mixture thereof that does not interact with the starting materials, reagents, intermediates or products in a manner that adversely affects the yield of the desired product.
By "pharmaceutically acceptable" it is meant that the carrier, excipient (vehicle) or diluent and/or salt must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
The term "pharmaceutically effective amount" as used herein means an amount of a compound of formula I or IA (or a combination of a compound of formula I or IA and a combination) sufficient to treat, prevent or delay or reduce the symptoms and physiological manifestations of the indications described herein.
The term "room temperature" means a temperature between 18 and 25 ℃, the term "HPLC" means high pressure liquid chromatography, "MPLC" means medium pressure liquid chromatography, "TLC" means thin layer chromatography, "MS" means mass spectrometry or mass spectrometry, "NMR" means nuclear magnetic resonance spectroscopy, "DCM" means dichloromethane, "DMSO" means dimethylsulfoxide, "DME" means dimethoxyethane, "EtOAc" means ethyl acetate, "MeOH" means methanol, "Ph" means phenyl, "Pr" means propyl, "trityl" means trityl, "ACN" means acetonitrile, "DEAD" means diethyl azodicarboxylate, and "DIAD" means diisopropyl azodicarboxylate.
It is to be understood that if a carbocyclic or heterocyclic moiety can be bonded or otherwise attached to a given substrate via different ring atoms without indicating a specific point of attachment, all possible points are meant, whether via a carbon atom or, for example, a trivalent nitrogen atom. For example, the term "pyridyl" means 2-, 3-, or 4-pyridyl, the term "thienyl" means 2-or 3-thienyl, and the like. In general, the compounds of the invention can be prepared by methods that include methods analogous to those known in the chemical arts, especially in accordance with the description contained herein.
The term mono-N-or di-N, N- (C) as used herein1-Cx) Alkyl … … means when it is di-N, N- (C)1-Cx) Alkyl … …, the above (C)1-Cx) Alkyl moieties are independently employed (x means an integer).
Halogen means chlorine, bromine, iodine or fluorine.
Alkyl means a straight chain saturated hydrocarbon or a branched chain saturated hydrocarbon. Examples of such alkyl groups (assuming the specified length includes specific examples) are methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, tert-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, hexyl, isohexyl, heptyl and octyl.
Alkoxy means a straight-chain saturated alkyl group or a branched-chain saturated alkyl group bonded via an oxy group. Examples of such alkoxy groups (assuming that the specified length includes specific examples) are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentyloxy, isopentyloxy, neopentyloxy, tert-pentyloxy, hexyloxy, isohexyloxy, heptyloxy and octyloxy.
The following sections describe exemplary rings for use in the general ring description contained herein.
Exemplary five to six membered aromatic rings optionally having one to three heteroatoms independently selected from oxygen, nitrogen and sulfur include phenyl, furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrimidinyl and pyrazinyl.
Exemplary six-membered rings include 2H-pyranyl, 4H-pyranyl, pyridyl, piperidyl, 1, 2-dioxinyl, 1, 3-dioxinyl, 1, 4-dioxanyl, morpholinyl, 1, 4-dithianyl, thiomorpholinyl, pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, 1,3, 5-triazinyl, 1,2, 4-triazinyl, 1,2, 3-triazinyl, 1,3, 5-trithianyl, 4H-1, 2-oxazinyl, 2H-1, 3-oxazinyl, 6H-1, 2-oxazinyl, 1, 4-oxazinyl, 2H-1, 2-oxazinyl, 4H-1, 4-oxazinyl, 1,2, 5-oxazinyl, 2H-1, 3-oxazinyl, 2H-1, 3-oxazinyl, 2,3-, 1, 4-oxazinyl, o-isooxazinyl, p-isooxazinyl, 1,2, 5-oxathiazinyl, 1,2, 6-oxathiazinyl, 1,4, 2-oxadiazinyl, and 1,3,5, 2-oxadiazinyl.
Exemplary bicyclic rings consisting of two fused partially saturated, fully saturated or fully unsaturated five or six membered rings independently, optionally having one to three heteroatoms independently selected from nitrogen, sulfur and oxygen include indolizinyl, indolyl, isoindolyl, 3H-indolyl, 1H-isoindolyl, indolinyl, cyclopenta (b) pyridyl, pyrano (3,4-b) pyrrolyl, benzofuranyl, isobenzofuranyl, benzo (b) thienyl, benzo (c) thienyl, 1H-indazolyl, indolizinyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, purinyl, 4H-quinolizinyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 1, 8-naphthyridinyl, pteridinyl, indenyl, indolinyl, cinnolinyl, quinoxalinyl, 1, 8-naphthyridinyl, indolizinyl, cyclopentidinyl, cyclopentadiinyl, and cyclopentadiinyl, Isoindenyl, naphthyl, tetrahydronaphthyl, naphthylalkyl, 2H-1-benzopyranyl, pyrido (3,4-b) -pyridinyl, pyrido (3,2-b) -pyridinyl, pyrido (4,3-b) -pyridinyl, 2H-1, 3-benzoxazinyl, 2H-1, 4-benzoxazinyl, 1H-2, 3-benzoxazinyl, 4H-3, 1-benzoxazinyl, 2H-1, 2-benzoxazinyl and 4H-1, 4-benzoxazinyl.
Certain methods of preparing the compounds of the invention are provided as further features of the invention and are illustrated by the following reaction schemes. Other methods can be illustrated in the experimental section. Specific synthetic reaction schemes for preparing compounds of formula I or IA are outlined below.
As an initial note in the preparation of compounds of formula I or IA, it is noted that some preparation methods useful for preparing the compounds described herein may require protection of remote functional groups (e.g., primary amine, secondary amine, carboxyl groups in precursors of formula I or IA). The need for such protection will vary depending on the nature of the remote functionality and the conditions of the preparation process. The need for such protection is readily determined by one skilled in the art. The use of such protection/deprotection methods is also within the skill of the art. For a general description of protecting Groups and their use, see T.W. Greene, Protective Groups in organic Synthesis, John Wiley & Sons, New York, 1991.
For example, certain primary amine or carboxylic acid functional groups of certain compounds, if left unprotected, may interfere with reactions elsewhere in the molecule. Thus, these functional groups may be protected by suitable protecting groups and the protecting groups may be removed in a subsequent step. Suitable protecting groups for protecting amines and carboxylic acids include those commonly used in peptide synthesis (such as N-tert-butoxycarbonyl, benzyloxycarbonyl, and 9-fluorenylmethyleneoxycarbonyl for amine protection and lower alkyl or benzyl esters for carboxylic acid protection), which are generally unreactive under the chemical conditions described and which are generally removable without chemically altering the other functional groups of the compounds of formula I or IA.
Reaction scheme I
Those skilled in the art are aware of the various methods for preparing thiouracil, including the condensation of thiourea with various carbonyl-containing compounds, or by reacting uracil with thiohybridizing agents such as phosphorus pentasulfide or Lawesson's reagent. In forming the thiouracils of the present invention, it is useful to consider construction methods to enhance the desired compound in a regiospecific manner among the various possible isomers.
Thiouridine compounds of formula I or IA (wherein R is1And R2As described above) can be prepared from the enamines of formula III by a cyclization reaction. The enamine of formula III is cyclized to the corresponding thiouracil of formula I or IA, for example, by reaction with an isothiocyanate such as benzyl isothiocyanate, carboxyethyl isothiocyanate, or preferably (trimethylsilyl) isothiocyanate (TMSNCS). The reaction is under reaction conditions such as polarAn aprotic solvent (e.g., methyltetrahydrofuran, tetrahydrofuran, dioxane, isobutyronitrile, or in simple isothiocyanates) at about 20 ℃ to about 150 ℃, typically about 85 ℃ (by microwave or thermal heating methods) for about three hours to about forty-eight hours.
The enamine of formula III may conveniently be prepared from the β -ketoester of formula VI by reaction with the appropriate R2-NH2Amine (wherein R2As described above or wherein in R2In (A) is a suitably protected form such as O-tert-butyl carbamate, e.g., β -ketoester of formula VI is reacted with R2-NH2The amine is reacted in a weak acid such as acetic acid, in a polar solvent (e.g., methanol, ethanol, isopropanol, toluene, or simply in an amine), at about 20 ℃ to about 120 ℃ for about four hours to about seventy-two hours, typically about 80 ℃ for about 12 hours.
As described above, the β -ketoester of formula VI can be prepared, for example, from a methyl ketone of formula X, a carboxylic acid of formula XV, an aryl halide of formula XX, or other precursors known to those skilled in the art.
The beta-ketoesters of formula VI are prepared from methyl ketones of formula X by an alkoxycarbonylation (carboalkoxylation) reaction. For example, a methyl ketone of formula X is reacted with a dialkyl carbonate (preferably dimethyl carbonate) in the presence of an alkoxide base such as potassium tert-butoxide, in a polar solvent such as methyl tert-butyl ether or the corresponding alcohol for dialkyl carbonate, at about 15 ℃ to about 100 ℃, typically at room temperature for about four hours to about forty-eight hours, typically twelve hours.
The β -ketoesters of formula VI can also be prepared, for example, from carboxylic acids. For example, the β -ketoester of formula VI can be prepared from an activated carboxylic acid. The acid of formula XV is conveniently converted to the corresponding XVII acylimidazole by reaction with 1,1' -carbonyldiimidazole in a polar solvent, typically in tetrahydrofuran, at a temperature between 0 ℃ and 100 ℃, preferably at room temperature for 1 hour to twenty-four hours, preferably three hours. The conversion of the resulting solution of the acylimidazole XVII in a polar aprotic solvent such as tetrahydrofuran to the corresponding β -ketoester of formula VI is carried out by reaction with a solution of an activated acetate species such as the enolate of the acetate or preferably magnesium monoethyl malonate in a polar aprotic solvent such as tetrahydrofuran at a temperature between-80 ℃ and 100 ℃, preferably at room temperature, for one to forty-eight hours, preferably twelve hours, thereby preparing the corresponding β -ketoester of formula VI.
Those skilled in the art will recognize that there are various other methods available to prepare beta-ketoesters from acids.
It will be appreciated by those skilled in the art that the β -ketoesters of formula VI can also be prepared from esters of carboxylic acids of formula XV, such as methyl, ethyl, isopropyl or tert-butyl esters of carboxylic acids of formula XV, preferably isopropyl esters, by condensation reaction with an activated acetate species, such as the enolate of an acetate ester, preferably the enolate of isopropyl acetate, in a polar aprotic solvent such as tetrahydrofuran, dioxane or toluene, preferably tetrahydrofuran, at a temperature between-80 ℃ and 40 ℃, preferably at room temperature, for one to twenty four hours, preferably twelve hours, i.e. to prepare the corresponding β -ketoester of formula VI.
In addition, those skilled in the art are aware of various methods for converting aryl halides to β -ketoesters, including the following exemplary steps. Aryl halides (e.g., aryl bromides) of formula XX are combined with β -alkoxy acrylates of formula XXII such as ethyl 3-ethoxyacrylate by a palladium mediated coupling reaction using a palladium catalyst, typically bis (tri-tert-butylphosphine) palladium (optionally together with lithium chloride), in the presence of an amine such as N, N-dicyclohexylmethylamine under an inert atmosphere such as nitrogen at about 90 ℃ to about 140 ℃, typically at about 110 ℃, for about four hours to about forty-eight hours, typically twelve hours. The resulting enol ether of XXV is converted to the corresponding β -keto ester of formula VI by treatment with an acid such as aqueous hydrochloric acid in a polar solvent (e.g., dichloromethane, methanol, acetic acid) at about 15 ℃ to about 40 ℃, typically at about room temperature for about thirty minutes to about six hours.
Alternatively, the enamines of formula III may be prepared from the propiolates of formula XXX by the following two-step reaction.
Of formula XXXIIAlkynes with boronic acids of formula XXXV (wherein R1As described above) to produce a propiolate of formula XXX by a transition metal mediated coupling reaction. For example, let R be appropriate1Boric acid is reacted with cesium carbonate, copper iodide, silver (I) oxide and tert-butyl acrylate in a polar aprotic solvent such as dichloromethane at a temperature of about 60 ℃ to about 100 ℃, typically at about 80 ℃ for about 30 minutes to about six hours.
Passing the resulting propiolate of formula XXX through the appropriate R2-NH2Amine (wherein R2As described above) in the presence of a weak acid such as acetic acid to convert to the corresponding enamine of formula III. The reaction is carried out in a polar solvent such as ethanol or isopropanol at a temperature of about 60 ℃ to about 100 ℃, typically about 80 ℃ for about 24 hours to about 72 hours.
Reaction scheme II
Thiouracils of formula I or IA can also be prepared from 6-halothiouracils of formula LVI as shown in reaction scheme II.
The thiocarbonyl group in the halosulfururacils of formula LVI is protected, for example, by reaction with methyl iodide in the presence of a base such as diisopropylethylamine in a polar solvent (e.g., acetonitrile) at a temperature of about 15 ℃ to about 40 ℃, typically at room temperature for about eight hours to about twenty-four hours. Reacting the resulting halide of the formula LVII with the appropriate R1-metal species (wherein R1As defined above) by reaction with [1,1' -bis (diphenylphosphino) ferrocene]Dichloropalladium (II) and R1Boronic acid (where R is1As described above) with 1, 4-dioxane and aqueous sodium carbonate. The mixture is heated at a temperature of about 90 ℃ to about 150 ℃, typically by subjecting to microwave irradiation at 120 ℃ for about fifteen minutes to about one hour. The resulting compound of formula LVIII is prepared by reaction with a nucleophile, typically by reaction with ammonium sulfide in a polar solventSuch as pyridine, at a temperature of from about 60 c to about 150 c, and deprotected, typically by microwave irradiation at 75 c for about fifteen minutes to about one hour, to produce a thiouracil of formula I or IA.
Halothiouracils of formula LVI can be prepared from the corresponding thiouracils of formula LV by, for example, a two-step deprotonation/lithium-halogen exchange with iodine. Thiouracil is typically treated with a base such as lithium diisopropylamide in a polar aprotic solvent such as tetrahydrofuran at a temperature of about-20 ℃ to about-100 ℃, typically at-78 ℃. The solution is then warmed to a temperature of from about 0 ℃ to about-25 ℃, typically-10 ℃ for from about fifteen minutes to about one hour to produce the corresponding lithium intermediate, followed by cooling to a temperature of from about-60 ℃ to about-80 ℃, typically-78 ℃, after which the lithium intermediate is reacted with iodine in a suitable polar aprotic solvent for from about 5 minutes to about forty-eight hours, typically eight hours.
Reaction scheme III
Thiouridine compounds of formula IB and LXI (wherein R1As described above and R3And R4Although not specifically indicated above, meaning generally the substituents described above) can be prepared from an enamine of the formula LX as shown in reaction scheme III. Thiouracils of formula LXI (wherein R is an alkyl group such as methyl, ethyl, isopropyl or tert-butyl, preferably methyl or ethyl) can be converted to the corresponding carboxylic acid, wherein R ═ H, by various methods known to those skilled in the art such as acid or base hydrolysis, preferably with 5 equivalents of sodium hydroxide or lithium hydroxide, in a polar solvent such as water, methanol, ethanol, tetrahydrofuran or mixtures of these solvents, preferably water and ethanol, at a temperature between 0 ℃ and about 100 ℃, preferably at room temperature for one hour to twenty-four hours, preferably four hours. The resulting carboxylic acids of formula LXI can be prepared by using amide coupling reagents known to those skilled in the art, such as propanephosphonic anhydride (T3P) or (CDI), preferably propanephosphineThe anhydride is converted to the amide of formula IB in the presence of an organic base such as pyridine, triethylamine, imidazole or diisopropylethylamine, preferably diisopropylethylamine, in a polar solvent such as N, N' -dimethylformamide, dichloromethane or ethyl acetate, preferably dichloromethane, at a temperature between 0 ℃ and reflux of the solvent, preferably at room temperature for a period of from 15 minutes to forty-eight hours, preferably eighteen hours.
Thiouracil compounds of formula LXI can be prepared from enamines of formula LX by reaction with an isothiocyanate, such as N-benzyl isothiocyanate, N-carboxyethyl isothiocyanate or preferably (trimethylsilyl) isothiocyanate (TMSNCS), optionally in a polar aprotic solvent such as methyltetrahydrofuran, tetrahydrofuran, dioxane, isobutyronitrile, N-butyl acetate, N' -dimethylformamide, preferably solely in the isothiocyanate, at a temperature between 20 ℃ and 150 ℃, typically at about 85 ℃ (heated with a microwave reactor or a conventional heat source) for 15 minutes to forty-eight hours, preferably three hours.
Reaction scheme IV
Thiouropyrimidines of formula IC, LXXI and LXXII can be prepared from thiouracils of formula LXX as shown in reaction scheme IV.
Thiouropyrimidines of the formula IC (wherein R1As described above and R3-R9Although not specifically indicated above, means substituents as described generally above and wherein R is6To R9At least one of which is bonded to the corresponding guanidine nitrogen via a carbonyl moiety) can be prepared from thiouracils of the formula LXXII (which can exist in various tautomeric forms such as those shown) by reacting a guanidine of LXXII with an acylating agent such as an acid chloride or an alkyl chloroformate known to those skilled in the art in the presence of an aqueous base solution such as sodium carbonate or sodium bicarbonate in a polar aprotic solvent such as tetrahydrofuran at a temperature between 0 ℃ and reflux of the solvent, preferablyOptionally at room temperature, to obtain the corresponding guanidine of formula IC. Alternatively, thiouracil of LXXII may be reacted with a dialkyl carbonate in the presence of an alkoxide base such as sodium ethoxide in a polar solvent such as the corresponding alcohol for the dialkyl carbonate at a temperature between 15 ℃ and about 100 ℃, preferably at 50 ℃ for four hours to forty-eight hours, preferably fifteen hours, to give the corresponding guanidine of formula IC.
Thiouropyrimidines of the formula LXXII can be prepared from the corresponding thiouracils of the formula LXX by reaction with a compound containing R7R8Guanidinating agents of NCN such as benzotriazole-R7R8N-azomethine, imidazole-R7R8N-azomethine or pyrazole-R7R8N-azomethine is prepared in a polar aprotic solvent, preferably N, N' -dimethylformamide, in the presence of a base, preferably diisopropylethylamine, at a temperature between 15 ℃ and 60 ℃, preferably at room temperature, for four to seventy-two hours, preferably eighteen hours. Alternatively, amines of formula LXX can be converted to activated thiouracil LXXI by reaction with an azomethine reagent such as 1, 1-bis (1H-benzotriazol-1-yl) azomethine, 1-bis (1H-imidazol-1-yl) azomethine or 1, 1-bis (1H-pyrazol-1-yl) azomethine, wherein X is a leaving group such as benzotriazole, imidazole, pyrazole, in the presence of a base such as diisopropylethylamine in a polar aprotic solvent such as N, N' -dimethylformamide at a temperature between 15 ℃ and 100 ℃, preferably at room temperature, for four hours to forty-eight hours, preferably eighteen hours. The resulting activated thiouracil of formula LXXI may be followed by R7R8Treatment of NH in a polar aprotic solvent such as N, N' -dimethylformamide in the presence of a base such as diisopropylethylamine at a temperature between 20 ℃ and 120 ℃, preferably at 60 ℃, for one hour to 24 hours, preferably three hours, affords the guanidinethionamide of formula LXXII.
The starting materials and reagents for the compounds of formula I or IA described above are also readily available or can be readily synthesized by those skilled in the art using conventional organic synthesis methods. For example, many of the compounds used herein are related to or derived from compounds that are of great scientific interest and are in commercial need, and thus many of such compounds are commercially available or reported in the literature or can be prepared from other commonly available materials by methods reported in the literature.
The cis/trans isomers can be separated by conventional techniques well known to those skilled in the art, such as chromatography and fractional crystallization.
Mixtures of stereoisomers may be separated by conventional techniques known to those skilled in the art. [ see, for example, E LEliel, "stereoschemistry of Organic Compounds" (Wiley, New York, 1994) ].
Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from appropriate optically pure precursors.
Alternatively, the racemate (or a racemic precursor) may be reacted with an appropriate optically active compound, for example an alcohol, or, if the compound of formula (I) contains an acidic or basic moiety, an acid or base such as tartaric acid or 1-phenylethylamine. The resulting mixture of diastereomers can be separated by chromatography and/or fractional crystallization, and one or both of the diastereomers can be converted to the corresponding pure enantiomers by methods well known to those skilled in the art.
The chiral compounds of the invention (and chiral precursors thereof) can be obtained in enantiomerically-enriched form using chromatography, typically HPLC, on a resin with an asymmetric stationary phase and a mobile phase consisting of a hydrocarbon (typically heptane or hexane) containing 0 to 50% isopropanol (typically 2 to 20%) and 0-5% alkylamine (typically 0.1% diethylamine). Concentrating the eluent to obtain an enriched mixture.
Pharmaceutically acceptable salts of compounds of formula I or IA can be prepared by one or more of three exemplary methods:
(i) by reacting a compound of formula I or IA with the desired acid or base;
(ii) removal of acid-or base-labile protecting groups in suitable precursors of compounds of formula I or IA or ring opening of suitable cyclic precursors such as lactones or lactams by use of a desired acid or base; or
(iii) One salt of a compound of formula I or IA is converted to the other by reaction with an appropriate acid or base or by means of an appropriate ion exchange column.
All three reactions are usually carried out in solution. The resulting salt may precipitate out and may be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization in the resulting salt can vary from fully ionized to almost unionized.
The compounds of the present invention may also be used in combination with other agents useful in the treatment of the diseases/conditions described herein, such as anti-atherosclerotic agents and anti-thrombotic agents.
Combined agent
The compounds of the present invention may be administered alone or in combination with one or more additional therapeutic agents. By "administered in combination" or "combination therapy" is meant that the compound of the invention and one or more additional therapeutic agents are administered concurrently to the mammal to be treated. When administered in combination, each component may be administered simultaneously or sequentially in any order at different points in time. Thus, each component may be administered individually but close enough in time to provide the desired therapeutic effect. Thus, the prophylactic and therapeutic methods described herein include the use of a combination of agents.
The combination is administered to the mammal in a therapeutically effective amount. By "therapeutically effective amount" is meant an amount of a compound of the present invention that is effective to treat a desired disease/condition, e.g., a cardiovascular condition such as acute coronary syndrome, when administered to a mammal, either alone or in combination with an additional therapeutic agent.
Additional therapeutic agents include anticoagulant or coagulation inhibitors, antiplatelet or platelet inhibitors, thrombin inhibitors, thrombolytic or fibrinolytic agents, antiarrhythmic agents, antihypertensive agents, calcium channel blockers (L-and T-forms), cardiac glycosides, diuretics (diruetics), mineralocorticoid receptor antagonists, NO-supplying agents such as organic nitrates, NO-promoting agents such as phosphodiesterase inhibitors, cholesterol/lipid lowering agents and lipid change therapies (lipid profile therapies), antidiabetic agents, antidepressant agents, anti-inflammatory agents (steroids and non-steroids), anti-osteoporosis agents, hormone replacement therapies, oral contraceptives, antiobesity agents, anxiolytic agents, antiproliferative agents, antineoplastic agents, antiulcer and gastroesophageal reflux agents, growth hormones and/or growth hormone secretagogues, thyroid mimetics (including thyroid hormone receptor antagonists), Anti-infective, antiviral, antibacterial and antifungal agents.
Including those used in the ICU region, such as dobutamine, dopamine, epinephrine (dpinepherine), nitroglycerin, nitroprusside (nitroprusside), and the like.
Including combinations useful in the treatment of vasculitis such as azathioprine, cyclophosphamide, mycophenolate mofetil, rituximab, and the like.
In another embodiment, the invention provides a combination wherein the second agent is at least one agent selected from the group consisting of a factor Xa inhibitor, an anticoagulant, an antiplatelet agent, a thrombin inhibitor, a thrombolytic agent, and a fibrinolytic agent.
Exemplary factor Xa inhibitors include apixaban and rivaroxaban.
Examples of suitable anticoagulants for use in combination with the compounds of the invention include heparin (e.g., unfractionated and low molecular weight heparins such as enoxaparin and dalteparin).
In another preferred embodiment, the second agent is at least one agent selected from warfarin, unfractionated heparin, low molecular weight heparin, synthetic pentasaccharide, hirudin, argatroban (argatroban), aspirin, ibuprofen, naproxen, sulindac, indomethacin, mefenamic acid (mefenamate), droxyxicam, diclofenac, sulpirenone, piroxicam, ticlopidine, clopidogrel, tirofiban, eptin, abciximab), melagatran, hirudin (disulfatohirudin), tissue plasminogen activator, modified tissue plasminogen activator, anistreplase, urokinase, and streptokinase.
The preferred second agent is at least one antiplatelet agent. Particularly preferred antiplatelet agents are aspirin and clopidogrel.
The term antiplatelet agent (or platelet inhibitor) as used herein means an agent that can inhibit platelet function, for example, by inhibiting platelet aggregation, adhesion, or granule secretion. Medicaments include, but are not limited to, various known non-steroidal anti-inflammatory drugs (NSAIDS) such as aspirin, ibuprofen, naproxen, sulindac, indomethacin, mefenamic acid (mefenamate), droxicam, diclofenac, sulpirenone, piroxicam, and pharmaceutically acceptable salts or prodrugs thereof. Of the non-steroidal anti-inflammatory drugs (NSAIDS), aspirin (acetylsalicylic acid or ASA) and COX-2 inhibitors such as celecoxib or piroxicam are preferred. Other suitable platelet inhibitors include IIb/IIIa antagonists (e.g., tirofiban, eptifibatide, and abciximab), thromboxane-a 2-receptor antagonists (e.g., ifetroban), thromboxane-a 2-synthetase inhibitors, PDE-III inhibitors (e.g., chaperone, dipyridamole), and pharmaceutically acceptable salts or prodrugs thereof.
The term antiplatelet agent (or platelet inhibitor), as used herein, is also intended to include ADP (adenosine diphosphate) receptor antagonists, preferably purinergic receptor P2Y1And P2Y12Antagonist of (1), with P2Y12The antagonist of (3) is more preferred. Preferred P2Y12The receptor antagonist comprises ticagrelor, prasugrel, ticlopidine and clopidogrel, and comprises pharmaceutically acceptable salts or prodrugs thereof. Clopidogrel is a more preferred agent. Ticlopidine and clopidogrel are also preferred compounds because they are known to be gentle to the gastrointestinal tract when used.
The term thrombin inhibitor (or antithrombin agent) as used herein means an inhibitor of the serine protease thrombin. By inhibiting thrombin, various thrombin-mediated processes such as thrombin-mediated platelet activation (i.e., for example, platelet aggregation, and/or granular secretion of plasminogen activator inhibitor-1 and/or serotonin) and/or fibrin formation are interrupted. Some thrombin inhibitors are known to those skilled in the art and such inhibitors are contemplated for use in combination with the present compounds. Such inhibitors include, but are not limited to, norarginine derivatives, boropeptides, heparin, hirudin, argatroban, and melagatran, including pharmaceutically acceptable salts and prodrugs thereof. Boroarginine derivatives and boropeptides include the N-acetyl group of boronic acids and peptide derivatives such as the C-terminal alpha-amino boronic acid derivatives of lysine, ornithine, arginine, homoarginine and their corresponding isothiouronium analogs. The term hirudin as used herein includes suitable derivatives or analogues of hirudin, referred to herein as hirudins, such as hirudins (disturbithiodins). The term thrombolytic or fibrinolytic agent as used herein means an agent that dissolves blood clots (thrombi). These agents include tissue plasminogen activator (natural or recombinant) and modified forms thereof, anistreplase, urokinase, streptokinase, Tenecteplase (TNK), lanoteplase (nPA), factor vila inhibitors, PAI-1 inhibitors (i.e., inactivators of tissue plasminogen activator inhibitors), alpha 2-antifibrinolytic enzyme inhibitors, and anisylated plasminogen streptokinase activator complex, including pharmaceutically acceptable salts or prodrugs thereof. The term anistreplase as used herein means an anisylated plasminogen streptokinase activator complex as described, for example, in EP028,489, the disclosure of which is incorporated herein by reference. The term urokinase as used herein is intended to mean both double-and single-chain urokinase, the latter also being referred to herein as prourokinase.
Examples of suitable antiarrhythmic agents include: class I agents (such as propafenone); class II agents (such as metoprolol, atenolol, carvaciolo, and propranolol); class III agents (such as sotalol, dofetilide, amiodarone, azimilide, and ibutilide); class IV agents (such as diltiazem and verapamil); k+Channel openers such as IAchInhibitors and IKurInhibitors (e.g. ofCompounds such as those described in WO 01/40231).
The compounds of the present invention may be used in combination with antihypertensive agents and such antihypertensive activity can be readily determined by those skilled in the art according to standard assays (e.g., blood pressure measurement). Examples of suitable antihypertensive agents include alpha-adrenergic blockers; a beta-adrenergic blocker; calcium channel blockers (e.g., diltiazem, verapamil, nifedipine, and amlodipine); vasodilators (e.g., hydralazine); diuretics (e.g., chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide, bendroflumethiazide, methylchlorothiazide, trichlorthiazide, polythiazide, benthiazide, ethacrynic acid, tennic acid (tricrynafen), chlorthalidone, torasemide, furosemide, misomine (musolimine), bumetanide, triamterin (triamtreene), amiloride, spironolactone); a renin inhibitor; ACE inhibitors (e.g., captopril, zofenopril, fosinopril, enalapril, cilazapril (ceranopril), cilazapril (cilazopril), delapril, pentopril, quinapril, ramipril, and lisinopril); AT-1 receptor antagonists (e.g., losartan, irbesartan, valsartan); ET receptor antagonists (e.g., sitaxsentan (sitaxsentan), atrasentan (atrsentan), and the compounds disclosed in U.S. patent nos. 5,612,359 and 6,043,265); dual ET/AII antagonists (e.g. compounds disclosed in WO 00/01389); neutral Endopeptidase (NEP) inhibitors; vasopeptidase inhibitors (dual NEP-ACE inhibitors) (e.g., gimeracil and nitrate)). An exemplary anti-angina agent is ivabradine.
Suitable calcium channel blockers (either L-or T-form) include diltiazem, verapamil, nifedipine and amlodipine and mibefradil (mybefradil).
Examples of suitable cardiac glycosides include digitalis and umbilicin.
In one embodiment, a compound of formula I or IA may be co-administered with one or more diuretics. Examples of suitable diuretics include (a) loop diuretics such as furosemide (such as LASIX)TM) Torasemide(s)Such as DEMADEXTM) Bumetanide (e.g., BUMEX)TM) And ethacrynic acid (such as EDECRIN)TM) (ii) a (b) Thiazide diuretics such as chlorothiazide (such as DIURIL)TM、ESIDRIXTMOr HYDRODIRILTM) Hydrochlorothiazide (such as MICROZIDE)TMOr ORETICTM) Benzylthiazine, hydrofluorothiazine (such as salouron)TM) Bendroflumethiazide, methylchlorothiazide, polythiazide, trichlorthiazide and indapamide such as LOZOLTM) (ii) a (c) Phthalimidine-type diuretics such as chlorthalidone (such as HYGROTON)TM) And metolazone (such as ZAROXOLYN)TM) (ii) a (d) Quinazoline-type diuretics such as quinethazone; and (e) potassium sparing diuretics such as triamterin (such as DYRENIUM)TM) And amiloride (such as MIDAMOR)TMOr MODURETICTM)。
In another embodiment, a compound of formula I or IA may be co-administered with a loop diuretic. In another embodiment, the loop diuretic is selected from furosemide and torasemide. In another embodiment, one or more compounds of formula I or IA may be co-administered with furosemide. In another embodiment, one or more compounds of formula I or IA may be co-administered with torasemide (which may optionally be controlled release or modified release torasemide).
In another embodiment, a compound of formula I or IA may be co-administered with a thiazide-type diuretic. In another embodiment, the thiazide-type diuretic is selected from chlorothiazide and hydrochlorothiazide. In another embodiment, one or more compounds of formula I or IA may be co-administered with chlorothiazide. In another embodiment, one or more compounds of formula I or IA may be co-administered with hydrochlorothiazide.
In another embodiment, one or more compounds of formula I or IA may be co-administered with a phthalimidine-type diuretic. In another embodiment, the phthalimidine-type diuretic is chlorthalidone.
Suitable combined mineralocorticoid receptor antagonists include spironolactone and eplerenone.
Examples of suitable combinations of phosphodiesterase inhibitors include: PDE III inhibitors (such as cilostazol); and PDE V inhibitors (such as sildenafil).
The compounds of the present invention may be used in combination with cholesterol-regulating agents (including cholesterol-lowering agents) such as lipase inhibitors, HMG-CoA reductase inhibitors, HMG-CoA synthase inhibitors, HMG-CoA reductase gene expression inhibitors, HMG-CoA synthase gene expression inhibitors, MTP/Apo B secretion inhibitors, CETP inhibitors, bile acid absorption inhibitors, cholesterol synthesis inhibitors, squalene synthase inhibitors, squalene epoxidase inhibitors, squalene cyclase inhibitors, combined squalene epoxidase/squalene cyclase inhibitors, fibric acids (fibrates), nicotinic acid, ion exchange resins, antioxidants, ACAT inhibitors or bile acid sequestrants or agents such as meprobamate.
Suitable cholesterol/lipid lowering agents and lipid change therapies include: HMG-CoA reductase inhibitors (e.g., pravastatin, lovastatin, atorvastatin, simvastatin, fluvastatin, NK-104 (also known as itavastatin or nivastatin (nisvastatin) or nivastatin (nisstatin)) and ZD-4522 (also known as rosuvastatin or atorvastatin (atavastatin) or visastatin (visstatin))); squalene synthetase inhibitors, fibric acids (fibrates), bile acid sequestrants (such as cholestyramine); an ACAT inhibitor; an MTP inhibitor; (ii) a lipoxygenase inhibitor; cholesterol absorption inhibitors; and cholesteryl ester transfer protein inhibitors.
Anti-inflammatory agents also include sPLA2 and lpPLA2 inhibitors (such as daradadine), 5LO inhibitors (such as adrenalon (atreluton)) and IL-1r antagonists (such as canakinumab).
Other anti-atherosclerotic agents include agents that modulate the action of PCSK 9.
Cardiovascular complications of type 2 diabetes are associated with detrimental concentrations of MPO, and thus the compounds of the present invention may be useful againstExamples of suitable antidiabetic agents include, for example, insulin, metformin (metfomin), DPPIV inhibitors, GLP-1 agonists, analogues and mimetics, SGLT1 and SGLT2 inhibitors, suitable antidiabetic agents include acetyl-CoA carboxylase- (ACC) inhibitors such as those described in WO2009144554, WO2003072197, WO 20091555 and WO2008065508, diacylglycerol O-acyltransferase 1(DGAT-1) inhibitors such as those described in WO09016462 and WO2010086820, AZD7687 or LCQ glycopeptide, diacylglycerol O-acyltransferase 2(DGAT-2) inhibitors, monoacylglycerol O-acyltransferase inhibitors, Phosphodiesterase (PDE) -10 inhibitors, AMPK activators, sulfonylureas (e.g., hexylurea, chlorpropamide, trypticamide, prolide, glibenclamide, pyrazine, glipizide (glipizide), gliclazide-10 inhibitors, gliclazide agonists such as gliclazide-3, gliclazide-mesylate, gliclazide (e), gliclazide-reductase inhibitors such as gliclazide-3, gliclazide (e), gliclazide-3), gliclazide (e.g., gliclazide) inhibitors, gliclazide (e.g. gliclazide), gliclazide) inhibitors, gliclazide (e.g. gliclazide), gliclazide) inhibitors, gliclazide (e.g. gliclazide) inhibitors, gliclazide), gliclazide (e.g. gliclazide) inhibitors, gliclazide (e.g. insulin-7, gliclazide) inhibitors, gliclazide (e.g. insulin-7, gliclazide (e.g. insulin-1-7-8, gliclazide), gliclazide (e.g. gliclazide), gliclazide (e.g. insulin-gliclazide), gliclazide (e.g. insulin-gliclazide), gliclazide (e.g. insulin-gliclazideAlbelutide, linatide, dolaglutide, semaglutide, NN-9924, TTP-054, inhibitors of protein tyrosine phosphatase-1B (PTP-1B) (e.g., Quinumine, ceterourone (Hyrtiosal) extract and the peptide fragment discovery today, as disclosed in Zhang, S., et al, Drug discovery today, 12(9/10),373-381 (2007)), SIRT-1 inhibitors (e.g., resveratrol, GSK2245840 or GSK184072), dipeptidyl peptidase IV (DPP-IV) inhibitors (e.g., those in WO2005116014, sitagliptin, vildagliptin, alogliptin, dulagliptin, linagliptin and saxagliptin), insulin secretagogues, fatty acid oxidation inhibitors, a2 antagonists, c-jun amino terminal kinase (JNK) inhibitors, glucokinase activators (GKa) such as those described in WO2010103437, WO2010103438, WO2010013161, WO2007122482, TTP-399, TTP-355, TTP-547, AZD1656, arjun 403, MK-0599, TAK-329, AZD5658 or GKM-001, insulin, glycogen phosphorylase inhibitors (e.g. GSK 136282885), agonists of the receptor agonists such as described in sang-35. c.35,539) in WO 2007122432, those of 551-5,559 (month 7 2010) include dapagliflozin, canagliflozin, empagliflozin, tolfogliflozin (CSG 452), ASP-1941, THR1474, TS-071, ISIS388626 and LX4211 as well as those described in WO2010023594, glucagon receptor modulators such as those described in Demong, d.e.et al.anunalhealorts in Medicinal Chemistry2008, 43, 119-137, GPR119 modulators, especially agonists, such as those described in WO 0140092, WO2010128425, WO2010128414, WO 201010med6457, Jones, r.m.et al, pharmaceutical Chemistry2009, 149, such as those described in MBX-170, psnkx-22682, CSG 12982, CSG 12924, and gpseik 52, as agonists, especially gpseik receptor agonists, gpseik # 3652, gpseq.10 receptor modulators such as those described in gpseik r-5932, gpus r-369, gpus-55, pg7-55, pg14, gpus-55-20-k-WO-55, gpus-9, gpus-r-9, gpus-r-k-9, gpus-k-9, gpus-g-9-g-a-r-9-r-a-k-9, gpus-k-g-k-9, gpus-9-k-9-k-, 386-396 and INT777, GPR40 agonists such as those described in Medina, j.c., Annual Reports in Medicinal Chemistry2008, 43, 75-85, including but not limited to TAK875, GPR120 modulators, in particular agonists, high affinity nicotinic acid receptor (HM74A) activators, and SGLT1 inhibitors such as GSK 1614235. A further list of representative antidiabetic agents which may be combined with the compounds of the present invention may be found, for example, on page 28 line 35 of WO2011005611To page 30, line 19 preferred antidiabetic agents are metformin and DPP-IV inhibitors (e.g., sitagliptin, vildagliptin, alogliptin, dulagliptin, linagliptin and saxagliptin.) other antidiabetic agents may include inhibitors or modulators of carnitine palmitoyl transferase, inhibitors of fructose 1, 6-bisphosphatase, inhibitors of aldose reductase, mineralocorticoid receptor inhibitors, inhibitors of tofgarc 3, inhibitors of CCR2 and/or CCR5, inhibitors of PKC isoforms (e.g., PKC α, PKC β, PKC γ), inhibitors of fatty acid synthase, inhibitors of serine palmitoyl transferase, inhibitors of GPR81, GPR39, 43, GPR41, GPR105, kv1.3, retinol binding protein 4, glucocorticosteroid receptor, somatostatin receptor (e.g., SSTR1, antidiabetic r2, SSTR3 and SSTR5), modulators of sstk 582 or trp 5, tph 2, tph 5, rpk, rp.
One skilled in the art will recognize that the compounds of the present invention may also be used in conjunction with other cardiovascular or cerebrovascular therapies including PCI, stent, drug-coated stent, stem cell therapy, and medical devices such as implanted pacemakers, defibrillators, or cardiac resynchronization therapy.
Myeloperoxidase activity has been demonstrated in neuroinflammatory conditions, and therefore, the compounds of the present invention can be used in combination with neuroinflammatory and neurodegenerative agents in mammals. Examples of additional neuroinflammatory and neurodegenerative agents include antidepressants, antipsychotics, anti-pain agents, anti-alzheimer's disease agents, and anxiolytics. Examples of specific classes of antidepressants that may be used in combination with the compounds of the present invention include norepinephrine reuptake inhibitors, Selective Serotonin Reuptake Inhibitors (SSRIs), NK-1 receptor antagonists, monoamine oxidase inhibitors (MAOIs), reversible inhibitors of monoamine oxidase (RIMAs), Serotonin and Norepinephrine Reuptake Inhibitors (SNRIs), Corticotropin Releasing Factor (CRF) antagonists, and atypical antidepressants. Suitable norepinephrine reuptake inhibitors include tertiary amine tricyclic antidepressants (tricyclics) and secondary amine tricyclic antidepressants. Examples of suitable tertiary amine tricyclic antidepressants and secondary amine tricyclic antidepressants include amitriptyline, clomipramine, doxepin, imipramine, trimipramine, dothiepin, butiline, nortriptyline, protriptyline, amoxapine, desipramine, and maprotiline. Examples of suitable Selective Serotonin Reuptake Inhibitors (SSRIs) include fluoxetine, fluvoxamine, paroxetine and sertraline. Examples of suitable monoamine oxidase inhibitors include isocarboxazid, phenelzine and strong endocarp (cyclopramine). Examples of suitable reversible inhibitors of monoamine oxidase include moclobemide. Examples of suitable Serotonin and Norepinephrine Reuptake Inhibitors (SNRIs) for use in the present invention include venlafaxine. Examples of suitable atypical antidepressants include bupropion, lithium, trazodone, and viloxazine. Examples of anti-alzheimer's disease agents include NMDA receptor antagonists such as memantine; and cholinesterase inhibitors such as donepezil and galantamine. Examples of suitable classes of anxiolytic agents that may be used in combination with the compounds of the present invention include benzodiazepines and serotonin 1A receptor (5-HT1A) agonists and CRF antagonists. Suitable benzodiazepines include alprazolam, chlordiazepoxide, clonazepam, chloramine butyrate, diazepam, lorazepam, oxazepam and pramipepam. Suitable 5-HT1A receptor agonists include buspirone and ixabepilone. Suitable Corticotropin Releasing Factor (CRF) antagonists include verrucifont. Suitable atypical antipsychotics include paliperidone, ziprasidone, risperidone, aripiprazole, olanzapine, and quetiapine. Suitable nicotinic acetylcholine agonists include CP-601927 and Vancklan. Anti-pain agents include pregabalin), gabapentin, clonidine, neostigmine, baclofen, midazolam, ketamine, and ziconotide.
Especially when provided in single dosage units, there are potentially chemical interactions between the active ingredients of the combination. For this reason, when the compound of formula I or IA and the second therapeutic agent are combined in a single dosage unit, they are formulated such that, although the active ingredients are combined in a single dosage unit, physical contact between the active ingredients is minimized (i.e., reduced). For example, one active ingredient may be enterically coated. By enteric coating one of the active ingredients, it is possible not only to minimize contact between the combined active ingredients, but also to control the release of one of the components in the gastrointestinal tract such that one of the components is not released in the stomach but in the intestine. One of the active ingredients may also be coated with a material that can affect sustained release throughout the gastrointestinal tract and also serve to minimize physical contact between the combined active ingredients. Furthermore, the sustained release component may be further enteric coated so that release of the component occurs only in the intestine. Another approach involves the formation of a combination product in which one component is coated with a slow release and/or enteric release polymer and the other component is also coated with a polymer such as low viscosity grade hydroxypropyl methylcellulose (HPMC) or other materials known in the art to further separate the active ingredients. The polymer coating is used to form an additional barrier to interaction with other components.
There are also other ways of minimising contact between the components of the combination product of the invention, whether administered in a single dosage form or in separate forms but simultaneously administered in the same manner, which will be readily apparent to those skilled in the art once having the present specification.
In the treatment of combination therapy, the compounds of the present invention and other pharmaceutical treatments are administered to a mammal (e.g., human, male or female) by conventional methods.
The compounds of formula I or IA of the present invention, prodrugs thereof, and salts of the compounds and prodrugs are all suitable for medical use as agents for inhibiting myeloperoxidase in mammals, particularly humans, and are useful for treating various conditions (e.g., as described herein) in which such an effect is implicated.
Myeloperoxidase is believed to be involved in the pathological oxidation of proteins, lipids and nucleic acids and contributes to abnormal cholesterol metabolism, tissue destruction and organ dysfunction and may induce or contribute to the development of cardiovascular disease and associated adverse outcomes.
Diseases/conditions that may be treated according to the present invention include, but are not limited to, cardiovascular conditions, diabetes (e.g. type II diabetes) and diabetic complications, vascular conditions, neuroinflammatory conditions, neurodegenerative conditions, pain, cancer, sepsis, NASH (steatohepatitis), lung injury and hypertension, kidney disease and vasculitis syndromes, particularly with respect to ANCA (anti-neutrophil cytoplasmic antibodies), and the like.
Given the positive correlation between myeloperoxidase activation and the development of cardiovascular and related diseases/conditions, the compounds of formula I or IA of the present invention, prodrugs thereof, and salts of said compounds and prodrugs are useful for preventing, arresting and/or regressing atherosclerosis and its related disease states due to their pharmacological actions.
Myeloperoxidase (MPO) is believed to exhibit atherogenic biological activity during the progression of cardiovascular disease. Furthermore, MPO-producing oxidants have been observed to reduce the bioavailability of nitric oxide, an important vasodilator. Furthermore, MPO has been shown to play a role in plaque instability by causing activation of metalloproteinases, leading to weakening of the plaque fibrous cap and subsequent plaque instability and rupture. Given these broad range effects of MPO, MPO has been implicated in a wide variety of cardiovascular diseases.
Cardiovascular diseases include, but are not limited to, coronary heart disease, acute coronary syndrome, ischemic heart disease, first and recurrent myocardial infarction, secondary myocardial infarction, non-ST-segment elevated myocardial infarction or ST-segment elevated myocardial infarction, sudden ischemic death, transient ischemic attack, peripheral occlusive arterial disease, angina pectoris, atherosclerosis, hypertension, heart failure (such as congestive heart failure), diastolic dysfunction (such as left ventricular diastolic dysfunction, diastolic heart failure, and impaired diastolic filling), systolic dysfunction (such as systolic heart failure with decreased ejection fraction), atrial fibrillation, arrhythmia (ventricular), ischemia, hypertrophic cardiomyopathy, sudden cardiac death, myocardial and vascular fibrosis, impaired arterial compliance, myocardial necrotic lesions, vascular injury, left ventricular hypertrophy, myocardial infarction, myocardial, Decreased ejection fraction, heart damage, vessel wall hypertrophy, endothelial thickening, fibrinoid necrosis of coronary arteries, adverse remodeling, stroke, and the like. Also included are venous thrombosis, deep vein thrombosis, thrombophlebitis, arterial embolism, coronary thrombosis, cerebral arterial thrombosis, cerebral embolism, renal embolism, pulmonary embolism, and thrombosis resulting from (a) prosthetic valves or other implants, (b) indwelling catheters, (c) stents, (d) cardiopulmonary bypass, (e) hemodialysis, or (f) other steps that may contribute to thrombus that expose blood to an artificial surface. It is noted that thrombosis includes occlusion (e.g., after bypass surgery) and reocclusion (e.g., during or after percutaneous transluminal coronary angioplasty).
Cardiovascular complications of type 2 diabetes are associated with detrimental concentrations of MPO, and thus the compounds of the present invention may be used to treat diabetes and diabetic complications such as macrovascular disease, hyperglycemia, metabolic syndrome, impaired glucose tolerance, hyperuricemia, diabetes, cataracts, diabetic neuropathy, diabetic nephropathy, diabetic retinopathy, obesity, dyslipidemia (dysidimia), hypertension, hyperinsulinemia and insulin resistance syndrome.
Furthermore, the link between myeloperoxidase activity and disease has been demonstrated in neuroinflammatory and neurodegenerative conditions. Accordingly, the compounds of the present invention are particularly useful in the treatment of neuroinflammatory and neurodegenerative conditions (i.e., disorders or diseases) such as multiple sclerosis in mammals, including humans; migraine headache; epilepsy; alzheimer's disease; parkinson's disease; brain damage; stroke; cerebrovascular diseases (including cerebral arteriosclerosis, cerebral amyloid angiopathy, hereditary cerebral hemorrhage and hypoxic ischemia of the brain); cognitive disorders (including amnesia, senile dementia, HIV-associated dementia, dementia associated with alzheimer's disease, dementia associated with huntington's disease, dementia with lewy bodies, vascular dementia, dementia associated with drugs, delirium and mild cognitive disorders); psychological deficits (including down syndrome and fragile X syndrome); sleep disorders (including hypersomnia, circadian rhythm sleep disorders, insomnia, parasomnia and sleep deprivation) and psychiatric disorders (such as anxiety disorders (including acute stress disorder, generalized anxiety disorder, social anxiety disorder, panic disorder, post-traumatic stress disorder and obsessive compulsive disorder), invasive disorders (including acute hallucinogenic mania), impulse control disorders (including compulsive gambling and intermittent manic disorder), mood disorders (including bipolar I disorder, bipolar II disorder, mania, mixed affective state, major depression, chronic depression, seasonal depression, psychotic depression and postpartum depression), psychomotor disorders, psychotic disorders (including schizophrenia, schizoaffective disorder, schizophreniform and delusional disorder), drug dependencies (including narcotic dependence, alcoholism, depression, seasonal depression, psychotic depression and postpartum depression), Amphetamine dependence, cocaine addiction, nicotine dependence, and drug withdrawal syndrome); eating disorders (including anorexia, bulimia, binge eating disorder, hyperphagia, and binge eating); and pediatric psychoses (including attention deficit disorder, attention deficit/hyperactivity disorder, conduct disorder, and autism) comprising administering to the mammal a therapeutically effective amount of a compound of formula I or IA or a pharmaceutically acceptable salt thereof.
Other inflammatory diseases or disorders such as asthma, chronic obstructive pulmonary disease, cystic fibrosis, idiopathic pulmonary fibrosis, acute respiratory distress syndrome, sinusitis, rhinitis, psoriasis, dermatitis, uveitis, gingivitis, atherosclerosis, inflammatory bowel disease, glomerular injury, liver fibrosis, sepsis, proctitis, rheumatoid arthritis, and inflammation associated with reperfusion injury, spinal cord injury, and tissue injury/scarring/adhesion/rejection.
The term "renal disorder caused by a contrast agent" includes contrast-agent-induced renal disorders after the step of using a contrast agent includes the steps of cardiac surgery, non-cardiac surgery, and transplant surgery. The renal disorders caused by contrast agents also include those in which patients are at risk of primary or secondary myocardial infarction, and which are caused by the use of contrast agents with enhanced imaging.
The use of the compounds of formula I or IA of the present invention, their prodrugs, and the salts of such compounds and prodrugs, as pharmaceutical agents for treating the above-mentioned diseases/conditions in mammals (e.g., humans, males, or females) is evidenced by the activity of the compounds of the present invention in conventional in vitro and in vivo assays described below. In vivo assays, including appropriate modifications within the skill of the art, can be used to determine the activity of other agents as well as the compounds of the invention. This assay also provides a means to compare the activities of the compounds of formula I or IA of this invention, their prodrugs, and the salts of such compounds and prodrugs (or other agents described herein) with each other and with the activities of other known compounds. These comparisons can be used to determine dosage values for treatment of these diseases in mammals, including humans.
The following protocols may of course be varied by the person skilled in the art.
MPO Amplex Red Activity assay
Metal Peroxidase (MPO) activity was measured by monitoring the formation of resorufin produced by the oxidation of Amplex Red (10-acetyl-3, 7-dihydroxyphenoxazine) (Invitrogen, Carlsbad, CA) by MPO (Gomes, Fernandes et al 2005). The assay mixture (100 μ L total volume) contained 50mM NaPi pH7.4, 150mM NaCl, 1mM DTPA (diethylene triamine pentaacetic acid), 2% DMSO, 2 μ M H2O2, 30 μ M Amplex Red and the reaction was started by the addition of 100pMMPO (purified from human multinucleated white blood cells and purchased from Calbiochem/EMD Biosciences, Gibbstown, NJ). All analyses were performed in 96 well, half area, black, unbound surface, polystyrene disks (Corning) and production of resorufin (excitation 530nm, emission 580nm) was monitored every 20 seconds on a Spectramax M2 microdisk luminometer (Molecular Devices, Palo Alto, CA) equipped with Softmax Pro software (Molecular Devices, Palo Alto, CA). The reaction used to determine the background response rate consisted of all assay components and 4. mu.L of 500 units/mL bovine catalase (Sigma) in 50mM KPi pH 7.0. The background rate was subtracted from each reaction progress curve. All data were analyzed in microsoft Excel and Kaleidagraph (Synergy software) using nonlinear regression analysis.
The efficacy of inhibitors on MPO is to be determined (k)inact/Ki) Curve fitting the initial 600s reaction to equation 1, where V0Initial rate (in RFU/s) and t time (in seconds) to obtain the first order rate constant (k) of enzyme inactivation at each inhibitor concentrationobs)。
Equation 1 is a variation of the standard equation for slow binding inhibition, where the steady state velocity (V)s) Is set to zero. Each kobsThe value is obtained by subtracting the k of the uninhibited reactionobsThe value was corrected for automatic inactivation of the enzyme. Then correcting kobsValue versus inhibitor concentration ([ I ]]) Plotting and fitting into equation 2
Wherein k isinactIs the maximum rate of deactivation and KiTo achieve inhibitor concentration half the maximum inactivation rate (Copeland 2005).
Tables 1 and (1A) below provide myeloperoxidase inhibitory activities obtained in the following examples according to the above-described analysis.
TABLE 1 MPO k of the examplesinact/Ki
| Example # | MPO kinact/Ki(1/s/M) |
| 1 | 11600 |
| 2 | 12500 |
| 3 | 29300 |
| 4 | 8790 |
| 5 | 1280 |
| 6 | 12900 |
| 7 | 10900 |
| 8 | 12100 |
| 9 | 15000 |
| 10 | 619 |
| 11 | 6880 |
| 12 | 5010 |
| 13 | 745 |
| 14 | 5310 |
| 15 | 5420 |
| 16 | 6510 |
| 17 | 4460 |
| Example # | MPO kinact/Ki(1/s/M) |
| 18 | 8280 |
| 19 | 1960 |
| 20 | 1330 |
| 21 | 4530 |
| 22 | 6340 |
| 23 | 991 |
| 24 | 7640 |
| 25 | 9210 |
| 26 | 3180 |
| 27 | 15800 |
| 28 | 13700 |
| 29 | 14900 |
| 30 | 5870 |
| 31 | 7680 |
| 32 | 2120 |
| 33 | 4520 |
| 34 | 13100 |
| 35 | 4110 |
| 36 | 5980 |
| 37 | 6460 |
| 38 | 5240 |
| 39 | 4520 |
| 40 | 6430 |
| 41 | 5000 |
| 42 | 6070 |
| 43 | 34600 |
| 44 | 3440 |
| 45 | 4000 |
| 46 | 1110 |
| 47 | 2340 |
| 48 | 6570 |
| 49 | 2230 |
| 50 | 2610 |
| Example # | MPO kinact/Ki(1/s/M) |
| 51 | 6180 |
| 52 | 33.4 |
| 53 | 3290 |
| 54 | 7070 |
| 55 | 7740 |
| 56 | 4770 |
| 57 | 13100 |
| 58 | 11700 |
| 59 | 8480 |
| 60 | 3470 |
| 61 | 3530 |
| 62 | 6930 |
| 63 | 12200 |
| 64 | 22500 |
| 65 | 7940 |
| 66 | 1580 |
| 67 | 7520 |
| 68 | 1160 |
| 69 | 4250 |
| 70 | 1590 |
| 71 | 3570 |
| 72 | 3580 |
| 73 | 9870 |
| 74 | 14400 |
| 75 | 2040 |
| 76 | 4190 |
| 77 | 6660 |
| 78 | 9730 |
| 79 | 1580 |
| 80 | 4130 |
| 81 | 24300 |
| 82 | 3390 |
| 83 | 3510 |
| Example # | MPO kinact/Ki(1/s/M) |
| 84 | 6630 |
| 85 | 10700 |
| 86 | 3960 |
| 87 | 15400 |
| 88 | 898 |
| 89 | 276 |
| 90 | 11600 |
| 91 | 9360 |
| 92 | 22100 |
| 93 | 5120 |
| 94 | 6930 |
| 95 | 15400 |
| 96 | 2200 |
| 97 | 6310 |
| 98 | 1870 |
| 99 | 2920 |
| 100 | 16100 |
| 101 | 4140 |
| 102 | 4200 |
| 103 | 41800 |
| 104 | 1210 |
| 105 | 27300 |
| 106 | 10500 |
| 107 | 1280 |
| 108 | 5800 |
| 109 | 914 |
| 110 | 864 |
| 111 | 10300 |
| 112 | 14900 |
| 113 | 25900 |
| 114 | 9770 |
| 115 | 4380 |
| 116 | 9920 |
| Example # | MPO kinact/Ki(1/s/M) |
| 117 | 11400 |
| 118 | 5500 |
| 119 | 25600 |
| 120 | 9720 |
| 121 | 15800 |
| 122 | 9310 |
| 123 | 3780 |
| 124 | 4610 |
| 125 | 10600 |
| 126 | 18200 |
| 127 | 5810 |
| 128 | 7680 |
| 129 | 18700 |
| 130 | 4830 |
| 131 | 14500 |
| 132 | 3840 |
| 133 | 15300 |
| 134 | 5350 |
| 135 | 6750 |
| 136 | 1920 |
| 137 | 701 |
| 138 | 4530 |
| 139 | 2890 |
| 140 | 10400 |
| 141 | 10500 |
| 142 | 4210 |
| 143 | 8110 |
| 144 | 6010 |
| 145 | 5080 |
| 146 | 8950 |
| 147 | 6500 |
| 148 | 6690 |
| 149 | 9770 |
| Example # | MPO kinact/Ki(1/s/M) |
| 150 | 8970 |
| 151 | 3740 |
| 152 | 4770 |
| 153 | 2200 |
| 154 | 1070 |
| 155 | 8090 |
| 156 | 16800 |
| 157 | 7320 |
| 158 | 1750 |
| 159 | 11400 |
| 160 | 7540 |
| 161 | 29600 |
| 162 | 8950 |
| 163 | 8090 |
| 164 | 14900 |
| 165 | 1280 |
| 166 | 8920 |
| 167 | 20300 |
| 168 | 8890 |
| 169 | 17800 |
| 170 | 4030 |
| 171 | 8590 |
| 172 | 2950 |
| 173 | 2910 |
| 174 | 10500 |
| 175 | 459 |
| 176 | 2160 |
| 177 | 5130 |
| 178 | 11100 |
| 179 | 2790 |
| 180 | 6960 |
| 181 | 7160 |
| 182 | 8200 |
| Example # | MPO kinact/Ki(1/s/M) |
| 183 | 4830 |
| 184 | 5970 |
| 185 | 9740 |
| 186 | 3930 |
| 187 | 5640 |
| 188 | 2180 |
| 189 | 2210 |
| 190 | 4090 |
| 191 | 14100 |
| 192 | 10800 |
| 193 | 458 |
| 194 | 2560 |
| 195 | 5350 |
| 196 | 5640 |
| 197 | 5650 |
| 198 | 8460 |
| 199 | 9080 |
| 200 | 4930 |
| 201 | 4350 |
| 202 | 8280 |
| 203 | 3450 |
| 204 | 3900 |
| 205 | 4900 |
| 206 | 7690 |
| 207 | 2400 |
| 208 | 3760 |
| 209 | 4360 |
| 210 | 968 |
| 211 | 6090 |
| 212 | 7590 |
| 213 | 4690 |
| 214 | 10700 |
| 215 | 1920 |
| Example # | MPO kinact/Ki(1/s/M) |
| 216 | 3260 |
| 217 | 3940 |
| 218 | 14100 |
| 219 | 1970 |
| 220 | 2420 |
| 221 | 6230 |
| 222 | 9820 |
| 223 | 3000 |
| 224 | 3280 |
| 225 | 5490 |
| 226 | 6280 |
| 227 | 745 |
| 228 | 615 |
| 229 | 5900 |
| 230 | 1740 |
| 231 | 1910 |
| 232 | 4520 |
| 233 | 2510 |
| 234 | 3060 |
| 235 | 2690 |
| 236 | 5740 |
| 237 | 2360 |
| 238 | 8740 |
| 239 | 1850 |
| 240 | 7070 |
| 241 | 7060 |
| 242 | 142 |
| 243 | 952 |
| 244 | 8970 |
| 245 | 1520 |
| 246 | 246 |
| 247 | 3060 |
| 248 | 3590 |
| Example # | MPO kinact/Ki(1/s/M) |
| 249 | 1050 |
| 250 | 7510 |
| 251 | 68 |
| 252 | 2480 |
| 253 | 12700 |
| 254 | 5630 |
| 255 | 3550 |
| 256 | 6520 |
| 257 | 3700 |
| 258 | 1460 |
| 259 | 4000 |
| 260 | 19700 |
| 261 | 2280 |
| 262 | 1730 |
| 263 | 4340 |
| 264 | 3620 |
| 265 | 3730 |
| 266 | 604 |
| 267 | 3840 |
| 268 | 6640 |
| 269 | 9510 |
| 270 | 20500 |
| 271 | 2010 |
| 272 | 3160 |
| 273 | 8180 |
| 274 | 22800 |
| 275 | 4730 |
| 276 | 6710 |
| 277 | 767 |
| 278 | 1560 |
| 279 | 386 |
| 280 | 430 |
| 281 | 1060 |
| Example # | MPO kinact/Ki(1/s/M) |
| 282 | 1180 |
| 283 | 2790 |
| 284 | 1470 |
| 285 | 1750 |
| 286 | 1500 |
| 287 | 2130 |
| 288 | 4230 |
| 289 | 1580 |
| 290 | 1890 |
| 291 | 2450 |
| 292 | 1070 |
| 293 | 1810 |
| 294 | 1910 |
| 295 | 793 |
| 296 | 1570 |
| 297 | 762 |
| 298 | 1080 |
| 299 | 2060 |
| 300 | 2460 |
| 301 | 3330 |
| 302 | 3630 |
| 303 | 5270 |
| 304 | 6290 |
| 305 | 6370 |
| 306 | 6740 |
| 307 | 14400 |
| 308 | 5340 |
| 309 | 3160 |
| 310 | 3110 |
| 311 | 2080 |
| 312 | 17100 |
| 313 | 973 |
| 314 | 429 |
| Example # | MPO kinact/Ki(1/s/M) |
| 315 | 1420 |
| 316 | 3060 |
| 317 | 7380 |
| 318 | 5240 |
| 319 | 7810 |
| 320 | 2390 |
| 321 | 2480 |
| 322 | 2800 |
| 323 | 10200 |
| 324 | 11300 |
| 325 | 1160 |
| 326 | 7480 |
| 327 | 1880 |
| 328 | 4370 |
| 329 | 963 |
| 330 | 5210 |
| 331 | 6330 |
| 332 | 3270 |
| 333 | 6100 |
| 334 | 6840 |
| 335 | 9820 |
| 336 | 589 |
| 337 | 13200 |
| 338 | 1280 |
| 339 | 10400 |
| 340 | 1450 |
| 341 | 14300 |
| 342 | 817 |
| 343 | 3570 |
| 344 | 8480 |
| 345 | 946 |
| 346 | 5890 |
| 347 | 378 |
| Example # | MPO kinact/Ki(1/s/M) |
| 348 | 1400 |
Table 1A below provides the myeloperoxidase inhibitory activities obtained in the following examples according to the above assay.
TABLE 1A. MPO k of the examplesinact/Ki
| Example # | MPO kinact/Ki(1/s/M) |
| 349 | 3630 |
| 350 | 8740 |
| 351 | 7870 |
| 352 | 6720 |
| 353 | 11000 |
| 354 | 1830 |
| 355 | 1540 |
| 356 | 2910 |
| 357 | 2940 |
| 358 | 1710 |
| 359 | 2660 |
| 360 | 2280 |
| 361 | 2060 |
| 362 | 2690 |
| 363 | 9680 |
| 364 | 6580 |
| 365 | 9290 |
| 366 | 13600 |
| 367 | 1340 |
| 368 | 3270 |
| 369 | 8040 |
| 370 | 9060 |
| 371 | 4570 |
| 372 | 6250 |
| 373 | 12800 |
| 374 | 4600 |
| 375 | 11300 |
| Example # | MPO kinact/Ki(1/s/M) |
| 376 | 7870 |
| 377 | 8770 |
| 378 | 5040 |
| 379 | 7370 |
| 380 | 4470 |
| 381 | 1970 |
| 382 | 2310 |
| 383 | 5230 |
| 384 | 2930 |
| 385 | 3530 |
| 386 | 4960 |
| 387 | 4720 |
| 388 | 8690 |
| 389 | 4910 |
| 390 | 6250 |
| 391 | 3480 |
| 392 | 5830 |
| 393 | 13600 |
| 394 | 4020 |
| 395 | 6980 |
| 396 | 10900 |
| 397 | 4050 |
| 398 | 4780 |
| 399 | 4860 |
| 400 | 2650 |
| 401 | 4060 |
| 402 | 4810 |
| 403 | 13300 |
| 404 | 6200 |
| 405 | 5970 |
| 406 | 4480 |
| 407 | 18700 |
| 408 | 9890 |
| Example # | MPO kinact/Ki(1/s/M) |
| 409 | 18000 |
| 410 | 3150 |
| 411 | 15000 |
| 412 | 3980 |
| 413 | 6560 |
| 414 | 1680 |
| 415 | 3910 |
| 416 | 4480 |
| 417 | 9280 |
| 418 | 11500 |
| 419 | 1200 |
| 420 | 5210 |
| 421 | 4950 |
| 422 | 4460 |
| 423 | 3290 |
| 424 | 6870 |
| 425 | 13400 |
| 426 | 4410 |
| 427 | 5360 |
| 428 | 5890 |
| 429 | 6620 |
| 430 | 9440 |
| 431 | 3440 |
| 432 | 1410 |
| 433 | 3490 |
| 434 | 4070 |
| 435 | 2420 |
| 436 | 3710 |
| 437 | 3400 |
| 438 | 7550 |
| 439 | 9200 |
| 440 | 3310 |
| 441 | 3260 |
| Example # | MPO kinact/Ki(1/s/M) |
| 442 | 12300 |
| 443 | 7330 |
| 444 | 17400 |
| 445 | 7350 |
| 446 | 14200 |
| 447 | 17200 |
| 448 | 6490 |
| 449 | 12000 |
| 450 | 7730 |
| 451 | 16000 |
| 452 | 11600 |
| 453 | 27800 |
TPO Amplex Red Activity assay
TPO Activity 2. mu. M H by the same assay as MPO2O230 μ M Amplex Red, and to obtain 1.3g of protein from HEK293 cell membranes expressing human TPO to initiate the reaction, cDNA encoding 933 amino acids in full length human TPO was cloned into inducible expression vector pcDNA5/frt/to (InVitrogen), 293 stable clones were selected using 100ug/mL hygromycin and 15ug/mL blasticidin (BLASTICIdine) in DMEM w/10% fetal calf serum 293 when the cells reached 50-60% confluence, expression of TPO was induced in medium containing above plus 10ug/mL doxycycline (doxycycline) and 5ug/mL hemin (Sigma) when the cells reached 50-60% confluence, the membranes were isolated from HEK293hTPO by harvesting in PBS, the cells were pelleted at 1000 g speed of 1000 g at 4 ℃ for 5min, resuspended in EDTA-free protease inhibitor (Roche) buffer (1mM, 7.7 mM in HEK293hTPO, the pellet was resuspended at 1000 ℃ for 5min and then assayed in pellet buffer containing 10mM Na + 20min, the pellet buffer (Invitrogen; the pellet) was homogenized at 20min, then assayed in pellet Na + 7mM buffer, the pellet was added to remove the pellet concentration of protease, homogenized protein, homogenized at 1000 mM temperature, then assayed in Tris buffer 20min, the pellet was added to obtain pellet, homogenized protein buffer containing EDTA-60% buffer, homogenized protein, dissolved pellet, and the pellet, dissolved pellet, assayed in Tris at 1000 min, then assayed in Tris buffer at 1000 g buffer at 1000 ℃ for 5min, then assayed at 1000 min, and the pellet, dissolved in PBS, and the pellet, then the pellet, assayed at 20min, dissolved in PBS, and the pellet, dissolved in the pellet, assayed at 150 min, the pellet.
IC50Values were measured by plotting the initial rate (the first 200 seconds of each reaction progress curve), expressed as a percentage of inhibition relative to the uninhibited (dimethylsulfoxide) reaction, as a function of inhibitor concentration. Fitting data to equation 3
Wherein the IC50Is the inhibitor concentration at 50% inhibition and z is the Hill slope (the slope of the curve at its inflection point).
Reference data
Copeland，R.A.(2005)。Evaluation of Enzyme Inhibitors in Drug DiscoveryA Guide for Medicinal Chemists and Pharmacologists.Hoboken，Wiley.
Gomes，A.，E.Fernandes，et al.，(2005)。“Fluorescence probes used fordetection of reactive oxygen species.”J Biochem Biophys Methods65(2-3)：45-80.
Human whole blood assay for irreversible MPO inhibition
To measure the inhibitory effect of MPO activity in the biological System of the present invention, whole blood collected from human volunteers without medical treatment was bioanalyzed in heparin-treated tubes (APP Pharmaceuticals, LLC, cat # NDC # 63323-. The blood was divided into aliquots and treated with different concentrations of MPO inhibitor or vehicle (vehicle) controls with or without bacterial lipopolysaccharide (LPS, InVivogen, cat # tlrl-pelps) to stimulate the simultaneous production of H by blood leukocytes2O2(an essential MPO substrate) and release MPO after 4 hours of incubation at room temperature, the plasma fraction was collected by centrifugation at 2000 × g at 4 ℃.
Plasma fractions were divided in two for analysis of total and active MPO. Total MPO amounts were determined using a standard sandwich ELISA assay (capture and detection antibodies: Cell Sciences, Cat # HP9048 and Cell Sciences, Cat # HM2164, clones 266-6K1) and calculated relative to a purified MPO (myeloperoxidase, Calbiochem, Cat #475911) standard curve (which was made by dilution in plasma from autologous donors). MPO activity was measured by capturing total MPO in plasma using a capture step as described for the ELISA method. After washing of the unbound plasma material, including unreacted MPO inhibitor, MPO-reactive substrates [ hydrogen peroxide (2uM) and Amplex Red (Invitrogen, Cat # a12222) ] were added and the Vmax of Amplex Red MPO substrate catalytically converted to resorufin was measured by measuring the increase in fluorescence in a kinetic assay using a fluorescence disc reader (excitation 530nm, emission 580 nm). The MPO activity of the capture material was compared to a standard curve of purified MPO (myeloperoxidase, Calbiochem, cat #475911) prepared in plasma from autologous donors. The percent "active" myeloperoxidase for each sample was calculated from the ratio of active myeloperoxidase in the Amplex Red assay to total myeloperoxidase obtained from ELISA for each sample. The dose response curves of MPOi concentration versus MPO activity were then plotted to determine IC50 values.
The compounds of the present invention may be administered via any method that delivers the compounds of the present invention systemically or locally. These methods include oral routes, parenteral, intraduodenal routes, buccal, intranasal, and the like. Typically, the compounds of the invention are administered orally, but when oral administration is not appropriate for the target or when the patient is unable to take the drug, then a parenteral route (e.g., intravenous, intramuscular, subcutaneous, or intramedullary) may be used.
For administration to a human patient, the oral daily dose of the compounds herein may range from 1mg to 5000mg, depending on, of course, the mode and frequency of administration, the disease state and age and condition of the patient, and the like. Oral daily doses in the range of 3mg to 2000mg may be administered. Further oral daily doses are in the range of 5mg to 1000 mg. For convenience, the compounds of the invention may be administered in unit dosage form. Multiple daily doses of the unit dosage form can be administered to increase the total daily dose, if desired. Unit dosage forms may be, for example, tablets or capsules containing about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 250, 500, or 1000mg of a compound of the invention. The total daily dose may be administered in a single dose or in divided doses and may fall outside the typical ranges provided herein at the discretion of the physician.
For administration to a human patient, the daily dose of the compound herein for infusion may range from 1mg to 2000mg, depending on, of course, the mode and frequency of administration, the disease state and age and condition of the patient, and the like. Further infusions were given daily doses in the range of 5mg to 1000 mg. The total daily dose may be administered in a single dose or in divided doses and may fall outside the typical ranges provided herein at the discretion of the physician.
These compounds may also be administered to animals other than humans, for example, for the indications detailed above. The precise dose of each active ingredient to be administered will vary depending on any factor including the type of animal and the type of disease state being treated, the age of the animal and the route of administration.
The dosage of the combination preparation used in combination with a compound of formula I or IA is effective for the indication to be treated. This dose can be determined by standard analysis such as those mentioned above and provided herein. The combination may be administered simultaneously or sequentially in any order.
These doses are based on an average human subject having about 60 to 70 kilograms. A physician will readily determine dosages for subjects with weights outside this range, such as infants and elderly.
The mode of administration may be adjusted to provide the most desirable desired response. For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be decreased or increased in proportion to the urgency of the medical condition. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein means physically discrete units suitable as unitary dosages for mammalian subjects; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The particulars of the dosage unit forms of the present invention are based upon and directly depend upon (a) the unique nature of the chemotherapeutic agent and the particular therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the art of synthesizing such active compounds for the treatment of susceptibility in individuals.
Thus, one skilled in the art will appreciate that, based on the description provided herein, the dosage and mode of administration are adjusted according to methods well known in the medical arts. That is, the maximum tolerable dose can be readily established, the effective amount to provide the patient perceptible medical benefit can be determined, and the time requirement for each agent to be administered to provide the patient perceptible medical benefit can also be determined. Thus, while certain dosages and modes of administration have been exemplified herein, these examples, of course, do not limit the dosages and modes of administration that can be provided to a patient in the practice of the present invention.
It is noted that dosage values may vary with the type and severity of the condition to be alleviated, and may include single or multiple doses. It will be further understood that the specific mode of administration for any particular subject must be adjusted over time according to the individual needs and the professional judgment of the person administering or supervising the administration of the compositions, and that the dosage ranges mentioned herein are exemplary only and are not intended to limit the scope or operation of the claimed compositions. For example, the dosage may be adjusted on the basis of pharmacokinetic or pharmacodynamic parameters, which include clinical effects such as toxicity profiles and/or laboratory values. Thus, the present invention encompasses the same patient dose escalation method as determined by one of skill in the art. Determining the appropriate dose and mode of administration for a chemotherapeutic agent is well known in the relevant art and, of course, is accomplished by those skilled in the art in light of the teachings disclosed herein.
The invention further comprises the use of a compound of formula I or IA as a medicament, such as a unit dose tablet or a unit dose capsule. In another embodiment, the invention encompasses the use of a compound of formula I or IA in the manufacture of a medicament (such as a unit dose tablet or unit dose capsule) for the treatment of one or more conditions previously identified in the treatment methods discussed above.
The pharmaceutical compositions of the present invention may be formulated (packaged or sold in bulk) as a single unit dose or as multiple single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition comprising a predetermined amount of active ingredient. The amount of active ingredient is generally equal to the dose to be administered to the subject or a convenient fraction of such a dose, such as half or one third of such a dose.
The compounds described herein may be administered in formulations comprising a pharmaceutically effective amount of a compound of formula I or IA, in combination with one or more pharmaceutically acceptable excipients (excipients) including carriers, excipients (vehicles) or diluents. The term "excipient" herein means any substance (not itself a therapeutic agent) that acts as a diluent, adjuvant or excipient (vehicle) for the delivery of a therapeutic agent to a subject or to be added to a pharmaceutical composition to improve its handling or storage properties or to allow or accelerate the formation of a solid dosage form such as a tablet, capsule or solution or suspension suitable for oral, parenteral, intradermal, subcutaneous or topical application. Excipients (excipients) may include, by way of illustration and not limitation, diluents, disintegrants, binders, adhesives, wetting agents, polymers, lubricants, glidants, stabilizers, substances added to mask or counteract unpleasant tastes and odors, flavors, dyes, fragrances, and substances added to improve the appearance of the composition. Acceptable excipients (excipients) include, but are not limited to, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, magnesium carbonate, talc, gelatin, acacia gum, sodium alginate, pectin, dextran, mannitol, sorbitol, lactose, sucrose, starch, gelatin, cellulosic materials such as cellulose esters and cellulose alkyl esters of alkanoic acids, low melting waxes, cocoa butter or powder, polymers such as polyvinyl-pyrrolidone, polyvinyl alcohol and polypropylene glycol, and other pharmaceutically acceptable materials. Examples of excipients (excipients) and their uses can be found in Remington's Pharmaceutical Sciences, 20th edition (Lippincott Williams & Wilkins, 2000). The choice of excipient (excipient) will depend to a large extent on factors such as the particular mode of administration, the effect of the excipient (excipient) on solubility and stability, and the nature of the dosage form.
The compounds herein can be formulated for oral, buccal, nasal, parenteral (e.g., intravenous, intramuscular, or subcutaneous), or rectal administration or in a form suitable for administration by inhalation. The compounds of the invention may also be formulated for sustained release.
Methods for preparing various pharmaceutical compositions containing certain amounts of active ingredients are known or will be apparent to those skilled in the art in light of the present specification. See Remington's pharmaceutical sciences, 20th Edition (Lippincott Williams & Wilkins, 2000).
The pharmaceutical compositions according to the invention may contain from 0.1% to 95% of the compounds according to the invention, preferably from 1% to 70%. In any event, the composition to be administered will contain an amount of a compound according to the invention effective to treat the disease/condition in the subject to be treated.
Since the present invention has an idea about treating the diseases/conditions described herein with a combination of active ingredients that can be administered individually, the present invention also relates to combining the individual pharmaceutical compositions in a kit (kit) form. The kit comprises two separate pharmaceutical compositions: a compound of formula I or IA, a prodrug thereof, or a salt of said compound or prodrug and a second compound as described above. The kit comprises means for containing the individual compositions such as containers, divided bottles, divided foil packets. Typically, the kit includes directions for administration of the individual components. The kit form is particularly advantageous when the individual components are preferably administered in different dosage forms (e.g. oral and parenteral), when administered at different dosing intervals, or when titration of the individual components of the combination is desired by the prescribing physician.
An example of such a kit is the so-called blister pack (blister pack). Blister packs are well known in the packaging industry and are widely used for the packaging of pharmaceutical unit dosage forms (tablets, capsules, etc.). Blister packs are typically composed of a sheet of relatively rigid material covered with a foil, preferably of transparent plastic material. During the packaging process, grooves are formed in the plastic foil. The recess has the size and shape of the tablet or capsule to be packaged. Thereafter, the tablet or capsule is placed in the recess and the sheet of relatively rigid material is sealed against the plastic foil on the side of the foil opposite to the direction in which the recess is formed. As a result, the tablet or capsule is sealed in the recess between the plastic foil and the sheet. Preferably, the strength of the sheet is such that the tablets or capsules can be removed from the blister pack by pressing by hand onto the grooves thereby forming openings in the sheet at the grooves. The tablet or capsule is then removed through the opening.
It may be desirable to provide memory aids on the kit, such as in the form of numbers next to the tablets or capsules that correspond to the number of days that the particular specified tablet or capsule must be taken. Another example of such a memory aid is a calendar printed on a card, such as "first week, Monday, Tuesday, etc. … … second week, Monday, Tuesday, … …", etc. Other variations of the memory aid will be readily apparent. The "daily dose" may be a single tablet or capsule or a few pills or capsules taken on a particular day. Also, the daily dose of a compound of formula I or IA may consist of one tablet or capsule, while the daily dose of a second compound may consist of several tablets or capsules and vice versa. The memory aid must react to this.
In another embodiment of the invention, a dispenser is provided that is designed to dispense daily doses one at a time for their intended use. Preferably, the dispenser is provided with memory aids to further assist compliance with the mode of administration. An example of such a memory aid is a mechanical counter, which displays the number of doses per day that have been dispensed. Another example of such a memory aid is a battery powered microchip having a liquid crystal readout or audible cue signal attached to the memory, which may, for example, read the date of the last daily dose that has been taken and/or remind of when the next dose was taken.
Furthermore, the present invention has an insight into treating the diseases/conditions described herein with a combination of active ingredients that can be co-administered, so the present invention also relates to combining individual pharmaceutical compositions in a single dosage form such as, but not limited to, a single tablet or capsule, a bi-or multi-layered tablet or capsule, or via the use of separate components or separate compartments within a tablet or capsule.
The active ingredient may be delivered in the form of a solution in an aqueous or non-aqueous vehicle (vehicle), with or without additional solvents, co-solvents, excipients (excipients) or a combination selected from pharmaceutically acceptable diluents, excipients (excipients), excipients (vehicles) or carriers.
An exemplary intravenous formulation is prepared as follows:
a solution of the above ingredients was administered intravenously to the patient at a rate of about 1mL per minute.
The active ingredient may be formulated as a solid dispersion with pharmaceutically acceptable excipients or as a Self Emulsifying Drug Delivery System (SEDDS).
The active ingredient may be formulated as immediate release or modified release tablets or capsules. In addition, the active ingredient can be delivered as the sole active ingredient within the capsule shell without additional excipients.
General Experimental procedure
All chemicals, reagents and solvents, when purchased, were purchased from commercial sources and used directly without further purification. Proton nuclear magnetic resonance spectrum (1H NMR) was recorded with a 400 and 500MHz Varian spectrometer. Chemical shifts are expressed in parts per million located in the tetramethylsilane low field. The peak shapes represent the following: s, singlet; d, double peak; t, triplet; q, quartet; m, multiplet; br s, broad singlet. Mass Spectrometry (MS) was performed by either Atmospheric Pressure Chemical Ionization (APCI) or Electron Scattering (ES) ionization sources. Unless noted otherwise, the observed masses (Obs Mass) reported in the tables correspond to the exact masses of the parent molecule plus one. Silica gel chromatography was mainly performed using medium pressure Biotage or ISCO systems, using columns pre-filled by various commercial suppliers including Biotage and ISCOThe process is carried out. Microanalysis was performed by Quantitative Technologies inc and was within 0.4% of the calculated values. The terms "concentration" and "evaporation" mean that the solvent is removed under reduced pressure on a rotary evaporator at a bath temperature of less than 60 ℃. The abbreviations "min" and "h" represent "minutes" and "hours", respectively.
Powder X-ray diffraction
Powder diffraction analysis was performed using a Bruker D8 diffractometer equipped with a copper (Cu) radiation source, a fixed slit (divergence 1.0mm, anti-scatter 0.6mm, and acceptance 0.6mm), and a scintillation counter detector data in a theta-theta goniometer at a copper wavelength K α 1 1.54056Next, collected from 2 θ angles of 3.0 to 40.0 using a diffraction step pitch (step size) of 0.040 degrees and a diffraction step time (step time) of 2.0 seconds. The X-ray tube voltage and amperage were set to 40kV and 40mA, respectively. Samples were prepared by placing in a Nickel Disk (Gasser)&Sons, inc. commac, NY) and rotated during data collection. Data were collected and analyzed using Bruker DIFFRAC Plus software (version 2.6).
I. Beta-ketoester route section
A. Carboxylic acid route moieties
Preparation example 1
3- (5-chloro-2-methoxyphenyl) -3-oxopropanoic acid ethyl ester
Magnesium ethoxide (67.46g, 589.51mmol) and tetrahydrofuran (1100mL) were charged into a 3000mL three-necked round bottom flask which had been purged with nitrogen, and the resulting mixture was stirred while ethyl hydrogen malonate (162.26g, 1.18 mol; 145.00mL diluted in 100mL tetrahydrofuran) was added, and the mixture was heated at 45 ℃ for 4 h. During this time, 5-chloro-2-methoxybenzoic acid (100g, 536mmol) and tetrahydrofuran (600mL) were charged to a 2000mL three-neck round bottom flask which had been purged with nitrogen. 1,1' -carbonyldiimidazole (95.59g, 589.5mmol) was added portionwise to this mixture being stirred at room temperature to avoid excessive foaming. After stirring at room temperature for 3h, the second solution was gradually added to the first solution. After the addition, the reaction mixture was heated to 45 ℃. After 20h, the reaction mixture was concentrated under reduced pressure, after which ethyl acetate (1L) followed by 2N hydrochloric acid (500mL) was added. After mixing, the layers were separated and the organic phase was washed successively with 2N hydrochloric acid (500mL), saturated sodium bicarbonate (500mL) and water (500 mL). The organic phase was concentrated under reduced pressure, and the residue was taken up in ethyl acetate (1000mL) and concentrated again to give the title compound (104.94 g).
MS(ES+)257.2[M+1]+。1H NMR showed the product to be a 7.5:1 mixture of keto-enols. In the case of keto tautomers:1H NMR(500MHz，CDCl3)7.85(d，J＝2.93Hz，1H)7.45(dd，J＝8.90，2.81Hz，1H)6.92(d，J＝8.78Hz，1H)4.18(q，J＝7.16Hz，2H)3.95(s，2H)3.90(s，3H)1.24(t，J＝7.07Hz，3H)。
preparation example 2
(Z) -3- ((2-amino-2-oxoethyl) amino) -3- (5-chloro-2-methoxyphenyl) propane
Ethyl enoate
Methanol (3.3L), sodium methoxide (102.4g, 1.8mol) and glycinamide hydrochloride (202g, 1.8mol) were charged to a 5L reaction vessel. The mixture was heated at 65 ℃ for 1h, then cooled to 50 ℃ and acetic acid (514.25mmol, 30.88g, 29.47mL) and ethyl 3- (5-chloro-2-methoxyphenyl) -3-oxopropanoate (300g, 1.03mol) were added. After heating to reflux for 16h, the reaction mixture was stirred while it was cooled to 10 ℃. After 30min, the resulting solid was collected by vacuum filtration, drained (pulling dry) to form a cake, which was dried in a vacuum oven (20mmHg, 65 ℃) for 14h to afford the title compound (339.4 g).
MS(ES+)313.2[M+1]+。1H NMR(500MHz，DMSO-d6)ppm8.80(t，J＝5.00Hz，1H)7.47(dd，J＝8.90，2.81Hz，1H)7.27(br.s.，1H)7.22(d，J＝2.68Hz，1H)7.14(d，J＝8.78Hz，1H)7.09(br.s.，1H)4.30(s，1H)4.03(q，J＝7.07Hz，2H)3.80(s，3H)3.56(br.s.，1H)3.45(br.s.，1H)1.18(t，J＝7.07Hz，3H)。
Example 1
2- (6- (5-chloro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide
Ethyl (Z) -3- ((2-amino-2-oxoethyl) amino) -3- (5-chloro-2-methoxyphenyl) acrylate (15g, 50.2mmol), butyl acetate (150mL), and trimethylsilyl isothiocyanate (160.7mmol, 21.1g, 22.7mL) were charged to a reaction vessel equipped with a high-efficiency stirrer, and the mixture was heated to reflux. After 15h, the mixture was cooled to 30 ℃ and treated with 1N aqueous sodium hydroxide (112.5mL, 112.5 mmol). After 30min, the organic layer was separated and extracted with another portion of 1N sodium hydroxide (37.5mL, 37.5 mmol). The combined aqueous phases were extracted twice with dichloromethane (2X 45mL), filtered and treated with 6N hydrochloric acid until a pH of 2.5 was reached. After stirring for 1h, the resulting solid was isolated by vacuum filtration, resuspended in 100mL1:1 methanol-water solution, heated at 50 ℃ for 2h with stirring, and then cooled to room temperature, after which the solid was collected by vacuum filtration, drained and dried in a vacuum oven (20mmHg, 50 ℃) for 12h to give 8.7g of the desired product as a tan solid.
MS(ES+)326.0[M+1]+。1H NMR(500MHz，DMSO-d6)ppm12.85(s，1H)7.57(dd，J＝9.03，2.68Hz，1H)7.33(s，1H)7.17-7.23(m，2H)7.10(s，1H)5.89(d，J＝1.71Hz，1H)5.41(br.s，1H)3.89(br.s，1H)3.84(s，3H)。
Another preparation method of example 1
2- (6- (5-chloro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide
A slurry of ethyl (Z) -3- ((2-amino-2-oxoethyl) amino) -3- (5-chloro-2-methoxyphenyl) acrylate (20g, 63mmol) in a mixture of butyl acetate (140mL) and N, N-dimethylformamide (38mL) was treated with trimethylsilyl isothiocyanate (16.8g, 125mmol) and the mixture was heated at 115 ℃ for 5-6 h. The mixture was cooled to 0-5 ℃, butyl acetate (100mL) was added and the mixture was slurried for 8 h. The solid formed was filtered and the filter cake was washed with butyl acetate (2X 100 mL). The solid was dried in a vacuum oven at 50 ℃ for 12h to a tan solid. The solid was dissolved in a mixture of 5:1N, N-dimethylformamide and water at room temperature and additional water was added slowly to crystallize the material. The slurry was cooled to 10 ℃ and stirred for a further 8h, after which it was filtered and washed with water. The filter cake was dried in a vacuum oven at 50 ℃ for 8 h. The solid was dissolved in a mixture of 1:1 methanol and water and the slurry was heated to 50 ℃ and held at this temperature for 2 h. After cooling to 10 ℃ during 30min, the slurry was kept at this temperature for 1h, filtered and washed with water and then dried in a vacuum oven at 50 ℃ for 8h to give the title compound as a white solid.
MS(ES+)326.0[M+1]+。1H NMR(500MHz，DMSO-d6)ppm12.85(s，1H)7.57(dd，J＝9.03，2.68Hz，1H)7.33(s，1H)7.17-7.23(m，2H)7.10(s，1H)5.89(d，J＝1.71Hz，1H)5.41(br.s，1H)3.89(br.s，1H)3.84(s，3H)。
Preparation example 3
1- (2, 5-Dimethoxyphenyl) -3-ethoxy-3-oxoprop-1-en-1-ol sodium
Magnesium ethoxide (3.61 mol; 413.52g) and tetrahydrofuran (6.6L) were charged into a 20L reaction vessel, and while ethyl hydrogen malonate (7.23 mol; 888.89 mL; 994.67g, diluted with 20mL tetrahydrofuran) was added, the resulting mixture was stirred and the mixture was heated at 45 ℃ for 4 h. During this time, 2, 5-dimethoxybenzoic acid (3.29 mol; 600.00g) and tetrahydrofuran (3.6L) were charged into a 20L reactor. 1,1' -carbonyldiimidazole (3.61 mol; 585.98g) was added portionwise to the mixture which was stirring at room temperature in order to avoid excessive foaming. After stirring at room temperature for 3h, the second solution was gradually added to the first solution. After the addition, the reaction mixture was heated to 45 ℃. After 20h, the reaction mixture was concentrated under reduced pressure, after which ethyl acetate (6L) followed by 2N hydrochloric acid (3L) was added. After mixing, the layers were separated and the organic phase was washed successively with 2N hydrochloric acid (3L), saturated sodium bicarbonate (3L) and water (3L). The organic phase is concentrated under reduced pressure, the residue is taken up in ethyl acetate (6L) and concentrated again to give an oil which is transferred with 5L of ethyl acetate into a 20L reaction vessel and treated with sodium methoxide (3.45 mol; 793.00mL of 4.35M solution in methanol). After stirring for 3h at room temperature, another 6L of ethyl acetate was added and the solid collected by vacuum filtration and then dried in a vacuum oven at 40 ℃ overnight to give 661g of the title product.
MS(ES+)253.1[M+1]+。1H NMR(400MHz，DMSO-d6)ppm6.92(d，J＝3.0Hz，1H)6.84(d，J＝8.8Hz，1H)6.73(dd，J＝8.8，3.0Hz，1H)4.67(s，1H)3.88(q，J＝7.0Hz，2H)3.67(s，6H)1.12(t，J＝7.0Hz，3H)。
Preparation example 4
(Z) -3- ((2-amino-2-oxoethyl) amino) -3- (2, 5-dimethoxyphenyl) acrylic acid ethyl ester
Methanol (3.3L), sodium methoxide (102.4g, 1.8mol) and glycinamide hydrochloride (202g, 1.8mol) were charged to a 5L reaction vessel. The mixture was heated at 65 ℃ for 1h, then cooled to 50 ℃ and acetic acid (514.25mmol, 30.88g, 29.47mL) and ethyl 3- (2, 5-dimethoxyphenyl) -3-oxopropanoate (300g, 1.03mol) were added. After heating at reflux for 16h, the reaction mixture was stirred while it was cooled to 10 ℃. After 30min, the resulting solid was collected by vacuum filtration, drained to form a cake, and dried in a vacuum oven (20mmHg, 65 ℃) for 14h to afford the title compound (339.4 g).
MS(ES+)309.1[M+1]+。1H NMR(400MHz，DMSO-d6)ppm8.84(t，J＝4.7Hz，1H)7.36(s，1H)7.09(s，1H)7.02(d，J＝8.9Hz，1H)6.97(dd，J＝8.9，2.8Hz，1H)6.74(d，J＝2.8Hz，1H)4.31(s，1H)4.03(q，J＝7.1Hz，2H)3.74(s，6H)3.58(br.s.，1H)3.47(br.s.，1H)1.18(t，J＝7.1Hz，3H)。
Example 2
2- (6- (2, 5-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide
Ethyl (Z) -3- ((2-amino-2-oxoethyl) amino) -3- (2, 5-dimethoxyphenyl) acrylate (1.30 mol; 400.00g), butyl acetate (3.4L) and trimethylsilyl isothiocyanate (4.15 mol; 585.67 mL; 544.96g) were charged to a 5L reaction vessel equipped with a high-efficiency stirrer, and the mixture was heated to reflux. After 16h, the mixture was cooled to 40 ℃ and treated with 2N aqueous sodium hydroxide (1.95L). The organic layer was separated and extracted with another portion of 2N sodium hydroxide (0.325L). The combined aqueous phases were filtered, extracted twice with dichloromethane (2X 1.6L) and slowly added at room temperature to a well stirred aqueous 3N hydrochloric acid solution (1.3L). After stirring for 30min, the resulting solid was isolated by vacuum filtration, washed with water and dried by suction to give a water-wet cake (640 g). The cake was dissolved in dimethylformamide (2.4L) at 90 deg.C and stirred while water (2L) was slowly added to the solution. The mixture was gradually cooled to room temperature, and the resulting solid was isolated by vacuum filtration, washed with water, and dried by suction to give 245g of a solid. The solid was then suspended in 1.25L of methanol and stirred while 1.25L of water was added. Heating the mixture at 50 deg.C for 2h while stirring, cooling to 10 deg.C for 2h, collecting the solid by vacuum filtration, draining, and drying in vacuum oven (20mmHg, 60 deg.C) to obtain the desired product.
MS(ES+)322.2[M+1]+。1H NMR(500MHz，DMSO-d6)ppm12.80(s，1H)7.32(br.s.，1H)7.06-7.11(m，2H)7.06(br.s.，1H)6.74-6.77(m，1H)5.82(d，J＝2.20Hz，1H)5.37(br.s.，1H)3.88(br.s.，1H)3.78(s，3H)3.70(s，3H)。
B. Methyl Ketone route section
Preparation example 5
3- (2, 4-Dimethoxyphenyl) -3-oxopropanoic acid methyl ester
A solution of potassium tert-butoxide (1M in tetrahydrofuran, 108.77mL, 108.77mmol), then 2, 4-dimethoxyacetophenone (10.00g, 54.38mmol) and dimethyl carbonate (13.93mL, 163.15mmol) in methyl tert-butyl ether (50mL) was added over 1.5h via an addition funnel to a three-necked round-bottomed flask under nitrogen equipped with a mechanical stirrer. During the addition, the reaction turned from an initially cloudy yellow mixture to a thick red-orange slurry. The reaction mixture was stirred at room temperature overnight. Aqueous citric acid (0.5N, 110.95mL, 54.39mmol) was added via the addition funnel to quench the reaction. An exothermic reaction was observed during the reaction termination and the solid dissolved to give an orange mixture. The layers were separated and the aqueous layer was extracted with methyl tert-butyl ether (2X 25 mL). The combined organic extracts were concentrated to low volume. Heptane (50mL) was added and a brown solid precipitated. The resulting slurry was stirred at room temperature under nitrogen overnight. The solid was filtered and dried under nitrogen to give the title compound as a beige powder (11.05g, 85% yield).
MS(ES+)239.1[M+1]+。1H NMR(500MHz，CDCl3)7.95(d，J＝8.78Hz，1H)6.57(dd，J＝8.78，2.20Hz，1H)6.45(d，J＝2.20Hz，1H)3.94(s，2H)3.88(s，3H)3.87(s，3H)3.73(s，3H)。
Preparation example 6
(Z) -3- (2, 4-Dimethoxyphenyl) -3- ((2-hydroxyethyl) amino) acrylic acid methyl ester
Ethanolamine (0.88mL, 14.69mmol) was added to a mixture of methyl 3- (2, 4-dimethoxyphenyl) -3-oxopropanoate (3.50g, 14.69mmol) and acetic acid (0.17mL, 2.94mmol) in isopropanol (70mL) and the reaction mixture was heated to 83 ℃. Additional ethanolamine (0.88mL, 14.69mmol) was added to the reaction mixture at 2,4, and 6 h. After stirring at 80 ℃ for 48h, the reaction mixture was cooled and concentrated under reduced pressure, after which the residue was suspended under nitrogen in equal parts of saturated sodium bicarbonate solution and water. After stirring overnight, the solid was collected by vacuum filtration and dried in a vacuum oven at 30 ℃ overnight to give the title compound as a beige powder (2.72g, 63%).
1H NMR(400MHz，CDCl3)8.77(t，J＝5.37Hz，1H)7.13(d，J＝8.29Hz，1H)6.47-6.52(m，2H)4.53(s，1H)3.84(s，3H)3.82(s，3H)3.66(s，3H)3.61(td，J＝5.45，5.45Hz，2H)3.15(td，J＝5.53，5.53Hz，2H)。
Example 3
6- (2, 4-Dimethoxyphenyl) -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one
(trimethylsilyl) isothiocyanate (23.80mL, 168.79mmol) was added to a solution of methyl (Z) -3- (2, 4-dimethoxyphenyl) -3- ((2-hydroxyethyl) amino) acrylate (9.50g, 33.77mmol) in 2-methyltetrahydrofuran (100mL) and the reaction mixture was heated at 85 ℃. After stirring overnight, the reaction mixture was cooled, extracted with 1N aqueous sodium hydroxide (1X 250mL then 1X 50mL), the combined aqueous layers were washed with dichloromethane (2X 50mL), and the aqueous phase was acidified to pH4 with concentrated HCl. The resulting solid was filtered, washed with water (2 × 50mL), and dried under nitrogen overnight to give a pale yellow powder. The product was dissolved in N, N-dimethylformamide (70mL) at 90 ℃ and then water (80mL) was added to this heated solution. After allowing to cool to room temperature and stirring overnight, the solid was collected by vacuum filtration, washed with water and dried under high vacuum to give the title compound as an off-white powder (6.7g, 61%).
MS(ES+)309.1[M+1]+。1H NMR(500MHz，DMSO-d6)ppm12.68(s，1H)7.24(d，J＝8.29Hz，1H)6.69(d，J＝2.44Hz，1H)6.65(dd，J＝8.42，2.32Hz，1H)5.70(d，J＝2.20Hz，1H)4.69(t，J＝4.88Hz，1H)4.50(ddd，J＝13.42，7.07，4.15Hz，1H)3.83(s，3H)3.82(s，3H)3.59(dt，J＝13.42，7.32Hz，1H)3.46-3.55(m，1H)3.38-3.46(m，1H)。
C. Aryl halide route moieties
Preparation example 7
(Z, E) -3- (2, 6-Dimethoxypyridin-3-yl) -3-ethoxyacrylic acid ethyl ester
Bis (tri-tert-butylphosphine) palladium (47mg, 0.092mmol) and lithium chloride (292mg, 0.27mmol) were added to a flask equipped with a reflux condenser, and the apparatus was evacuated in vacuo and refilled with nitrogen several times. Degassed anhydrous 1, 4-dioxane solution (8mL) was added to the flask under nitrogen through a cannula followed by 3-bromo-2, 6-dimethoxypyridine (500mg, 2.29mmol), N-dicyclohexylmethylamine (540 μ L, 2.52mmol) and ethyl 3-ethoxyacrylate (1.0mL, 6.88mmol) and the resulting orange solution was heated to 110 ℃. After 20h, the reaction mixture was cooled to room temperature, quenched with water and diluted with ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over sodium sulfate and concentrated in vacuo. The residue was purified by chromatography on silica gel eluting with 0-50% ethyl acetate/heptane to give the title compound as an amber oil (604mg, 94%).1HNMR showed the product to be composed of a 2.5:1 mixture of E/Z isomers.
Preparation example 8
3- (2, 6-Dimethoxypyridin-3-yl) -3-oxopropanoic acid ethyl ester
3N aqueous hydrochloric acid (3.5mL) was gradually added to a solution of ethyl 3- (2, 6-dimethoxypyridin-3-yl) -3-ethoxyacrylate (600mg, 2.13mmol) in dichloromethane (18 mL). The reaction mixture was stirred at room temperature for 2h, then carefully added to a saturated sodium bicarbonate solution (30 mL). The layers were separated and the aqueous layer was extracted with dichloromethane. The combined organic extracts were passed through a cotton plug to dry and concentrated in vacuo. The resulting oil was purified by flash chromatography (0-60% ethyl acetate/heptane) to afford the title compound as a white solid (515mg, 95% yield).
1H NMR(400MHz，CDCl3)ppm8.22(d，J＝8.59Hz，1H)6.40(d，J＝8.39Hz，1H)4.20(q，J＝7.03Hz，2H)4.03(s，3H)3.99(s，3H)3.94(s，2H)1.26(t，J＝7.13Hz，3H)。
D. Deprotection routes and derivatization of amines
Preparation example 9
(Z) -3- (2, 6-Dimethoxypyridin-3-yl) -3- ((2-hydroxyethyl) amino) acrylic acid ethyl ester
2-aminoethanol (0.60mL, 9.9mmol) followed by acetic acid (0.63mL, 9.9mmol) was added to a solution of ethyl 3- (2, 6-dimethoxypyridin-3-yl) -3-oxopropanoate (500mg, 1.97mmol) in ethanol (4 mL). The reaction mixture was heated to 90 ℃ for 16h, cooled to room temperature and concentrated in vacuo. The residue was partitioned between ethyl acetate and water. The organic layer was concentrated in vacuo and the crude material was purified by chromatography on silica gel eluting with 20-80% ethyl acetate/heptane to give the title compound as a clear gum (573mg, 98%).
MS(ES+)297.3[M+1]+。1H NMR(500MHz，CDCl3)8.75(br.s.，1H)7.44(d，J＝8.05Hz，1H)6.34(d，J＝8.05Hz，1H)4.51(s，1H)4.14(q，J＝7.16Hz，2H)3.97(s，3H)3.95(s，3H)3.64(td，J＝5.53，5.53Hz，2H)3.17(td，J＝5.53，5.53Hz，2H)1.96(br.s.，1H)1.27(t，J＝7.07Hz，3H)。
Example 4
6- (2, 6-Dimethoxypyridin-3-yl) -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one
(trimethylsilyl) isothiocyanate (0.30mL, 2.0mmol) was added to a solution of ethyl (Z) -3- (2, 6-dimethoxypyridin-3-yl) -3- ((2-hydroxyethyl) amino) acrylate (100mg, 0.34mmol) in 2-methyltetrahydrofuran (1.0mL) and the reaction mixture was heated at 80 ℃ for 4 h. The cooled reaction mixture was diluted with ethyl acetate and washed with saturated aqueous sodium bicarbonate. The layers were then separated and the organic layer was concentrated in vacuo. The residue was triturated with methanol and the resulting solid was collected by vacuum filtration to give the title compound as a white solid (16mg, 16%).
MS(ES+)310.2[M+1]+。1H NMR(500MHz，CD3OD)7.60(d，J＝8.29Hz，1H)6.47(d，J＝8.05Hz，1H)5.76(s，1H)4.66-4.75(m，1H)4.01(s，3H)3.98(s，3H)3.77-3.85(m，2H)3.57-3.63(m，1H)。
Preparation example 10
(Z) -3- (2- (tert-Butoxycarbonylamino) ethylamino) -3- (2, 4-dimethoxyphenyl) acrylic acid ethyl ester
A solution of ethyl 3- (2, 4-dimethoxyphenyl) -3-oxopropanoate (41.91g, 166mmol), tert-butyl 2-aminoethylcarbamate (54.7g, 342mmol) and acetic acid (16.14g, 269mmol) in ethanol (180mL) was heated at reflux for 5.3 h. After removal of most of the solvent by rotary evaporation, the resulting oil was partitioned between ethyl acetate (ca. 300mL) and 10% (w/v) aqueous ammonium chloride. The ethyl acetate layer was separated and washed with water, 10% (w/v) aqueous ammonium chloride (3mL), and brine (10 mL). The ethyl acetate layer was washed with saturated aqueous sodium bicarbonate, brine (6mL) was added, and the emulsion was allowed to settle. The ethyl acetate layer was finally washed with brine and dried over sodium sulfate. Evaporation of the volatile components of the ethyl acetate layer gave a viscous, amber toffee (62.3g, 95%). This crude product was used directly without further purification.
LCMS(ESI)m/z：395.4[M+H](100％)。1H NMR(500MHz，CDCl3)1.27(t，J＝7.1Hz，3H)，1.43(s，9H)，3.03-3.21(m，4H)，3.83(s，6H)，4.14(q，J＝7.1Hz，2H)，4.51(s，1H)，4.88(br.s.，1H)，6.47(d，J＝1.7Hz，1H)，6.50(dd，J＝8.4，1.8Hz，1H)，7.12(d，J＝8.3Hz，1H)，8.65(br.s.，1H)。
Example 5
2- (6- (2, 4-Dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethylcarbamic acid tert-butyl ester
(trimethylsilyl) isothiocyanate (66mL, 470mmol) was added to a solution of ethyl (Z) -3- (2- (tert-butoxycarbonylamino) ethylamino) -3- (2, 4-dimethoxyphenyl) acrylate (62.3g, 158mmol) in 2-methyltetrahydrofuran (160 mL). After heating under reflux under nitrogen for 15h, the reaction mixture was cooled to room temperature and quenched by the careful addition of saturated aqueous sodium bicarbonate (470 mL). The reaction mixture was extracted with dichloromethane, and the aqueous layer was extracted twice more with dichloromethane. The combined organic layers were dried over sodium sulfate and evaporated to give a yellow-amber foam which was purified by chromatography on silica gel eluting with 0-80% ethyl acetate in heptane to give 49.2g of a solid. These solids were resuspended in 1:1 ethyl acetate/heptane at 70 ℃ for 1h and then at room temperature for another 1 h. The resulting solid was isolated by vacuum filtration, rinsed with additional 1:1 ethyl acetate/heptane, and drained on the filter. The title compound was obtained as a white, microcrystalline solid (38.3g, 59.5% yield).
LCMS(ESI)m/z：408.3[M+H](100％)。1H NMR(500MHz，CDCl3Major rotamers) 1.40(s, 9H), 3.23-3.45(m, 2H), 3.74(dt, J ═ 14.4, 5.4Hz, 1H), 3.84(s, 3H), 3.87(s, 3H), 4.68-4.81(m, 2H), 5.81(d, J ═ 2.2Hz, 1H), 6.51(d, J ═ 2.2Hz, 1H), 6.59(dd, J ═ 8.4, 2.1Hz, 1H), 7.26(d, J ═ 8.4Hz, 1H), 9.58(br.s., 1H).
Example 6
1- (2-aminoethyl) -6- (2, 4-dimethoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one hydrochloride
Acetyl chloride (55mL, 770mmol) was added slowly over a period of 3min to a solution of ethanol (50mL, 860mmol) in ethyl acetate (390mL) that had been cooled in an ice/water bath. After 5min, the cooling bath was removed and after stirring for 45min, the solution was added to tert-butyl 2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethylcarbamate (31.7g, 77.8 mmol). After a period of time a suspension formed and after stirring for a further 5h, the solid was collected by vacuum filtration and rinsed with ethyl acetate. The solid was drained and further dried in vacuo to afford 26.6g (99.3%) of the desired product as a colorless solid.
LCMS(ESI)m/z：291.3[M-NH3+H](100％)，308.3[M+H](33％)，615.5[2M+H](2.3％)。1H NMR(500MHz，CD3OD)3.06(ddd，J＝12.9，7.8，5.9Hz，1H)，3.12(ddd，J＝12.9，7.7，6.4Hz，1H)，3.87(s，3H)，3.89(s，3H)，4.14(ddd，J＝14.0，7.8，5.9Hz，1H)，4.82(ddd，J＝14.0，7.7，6.4Hz，1H)，5.80(s，1H)，6.70(dd，J＝8.3，2.2Hz，1H)，6.73(d，J＝2.2Hz，1H)，7.27(d，J＝8.3Hz，1H)。
Example 7
2- (2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethyl) guanidine
Diisopropylethylamine (0.22mL, 1.3mmol) was added to a suspension of 1- (2-aminoethyl) -6- (2, 4-dimethoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one hydrochloride (181.6mg, 0.528mmol) (product of example 6) and 1H-pyrazole-1-carboxamidine hydrochloride (90.6mg, 0.618mmol) in N, N-dimethylformamide (0.55 mL). After heating at 55 ℃ for 1h, the reaction mixture was cooled to room temperature, diluted with ethanol (1.6mL), and the solid product was collected by vacuum filtration and washed with additional ethanol. The precipitated solid was resuspended in ethanol (2.1mL) for 3h at room temperature, after which it was collected again by vacuum filtration and rinsed with additional ethanol. Drying to obtain the desired product in colorless solid. The solubility data for this product is consistent with its zwitterionic form.
1H NMR (500MHz, 20% DCI with CD3OD +2 dripped in D2O) 3.31-3.37(m, 1H), 3.67(ddd, J ═ 14.8, 8.6, 5.9Hz, 1H), 3.88-3.99(m, 1H), 3.90(s, 6H), 4.66-4.77(m, 1H), 5.80 (c), (b), (s，1H)，6.69(d，J＝2.2Hz，1H)，6.71(dd，J＝8.3，2.2Hz，1H)，7.28(d，J＝8.3Hz，1H)。
2- (2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethyl) guanidine hydrochloride
The above reaction product (116.3mg, 0.333mmol) was suspended in dioxane and treated with 4.0M hydrochloric acid/dioxane solution (0.30mL, 1.2 mmol). After complete vortexing, the volatile components in the mixture were removed to give a white solid (130.6mg, 0.338 mmol). LCMS (ESI) m/z: 350.1[ M + H ] (100%).
Preparation example 11
2- (2- (6- (2, 4-Dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethylamino) -2-oxoethylcarbamic acid tert-butyl ester
O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (182mg, 0.48mmol) and tert-butoxycarbonylamino-acetic acid (70mg, 0.4mmol) and diisopropylethylamine (336mg, 1.6mmol) were added to a solution of 1- (2-aminoethyl) -6- (2, 4-dimethoxyphenyl) -2-thio-2, 3-dihydropyrimidin-4 (1H) -one hydrochloride (123mg, 0.4mmol) (product of example 6) in anhydrous dichloromethane (4 mL). After stirring at room temperature overnight, the reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative thin layer chromatography (1:1 petroleum ether: ethyl acetate) to give tert-butyl 2- (2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethylamino) -2-oxoethylcarbamate (120mg, 65%).
Example 8
2-amino-N- (2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethyl) acetamide hydrochloride
A solution of hydrochloric acid in ethyl acetate (2mL) was added to a solution of tert-butyl 2- (2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethylamino) -2-oxoethylcarbamate (70mg, 0.15mmol) in ethyl acetate (2 mL). After stirring at room temperature for 4H, the reaction mixture was concentrated under reduced pressure to give 2-amino-N- (2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) ethyl) acetamide hydrochloride (65mg, 100%) as a solid.
II.6-iodo-thiouracil route section
Preparation example 12
N- ((2-methoxyethyl) thiocarbamoyl) benzamide
2-Methoxyethylamine (17.7mL, 202.2mmol) was added dropwise over a period of 30min to a stirring solution of benzoyl isothiocyanate (30.00g, 183.8mmol) in dichloromethane (300mL) at room temperature under argon, and the mixture was stirred at room temperature for 16 h. The mixture was washed successively with 10% aqueous citric acid (75mL), water (75mL) and brine (75mL), dried over magnesium sulfate and concentrated in vacuo. The resulting yellow oil solidified upon standing to give the title compound (41.85g, 96%). This material was used directly in the next step without further purification.
Preparation example 13
1- (2-methoxyethyl) thiourea
A solution of N- ((2-methoxyethyl) thiocarbamoyl) benzamide (41.82g, 175.5mmol), potassium carbonate (24.25g, 175.5mmol) in methanol (200mL) and water (200mL) was stirred at room temperature for 16 h. The mixture was concentrated in vacuo and the aqueous layer was extracted with ethyl acetate (5X 100 mL). The combined organic layers were dried over sodium sulfate and concentrated in vacuo. The resulting yellow oil solidified upon standing to give the title compound (21.38g, 91%). This material was used directly in the next step without further purification.
MS(ES+)135.1(M+1)+。1H NMR(500MHz，CDCl3)6.66(br.s.，1H)6.46(br.s.，1H)5.81(br.s.，1H)3.80(br.s.，2H)3.48-3.65(m，2H)3.40(s，3H)。
Preparation example 14
1- (2-methoxyethyl) -2-thio-2, 3-dihydropyrimidin-4 (1H) -one
Freshly prepared 0.96N methanolic sodium methoxide solution (250mL, 239.0mmol) was added dropwise over a period of 30min to a stirring solution of 1- (2-methoxyethyl) thiourea (21.38g, 159.3mmol) and ethyl 3, 3-diethoxypropionate (46.5mL, 239.0mmol) in methanol (300mL) at room temperature under argon. The reaction mixture was heated to 60 ℃ for 45min and cooled to room temperature. The solvent was removed under reduced pressure and toluene (250mL) was added to the residue. The mixture was stirred at reflux for another 3h and then cooled to room temperature. Water (200mL) was added followed by separation of layers. The aqueous layer was washed with dichloromethane (50mL), neutralized with 2N aqueous hydrochloric acid, and extracted with dichloromethane (3X 200 mL). The combined organic layers were dried over sodium sulfate and concentrated in vacuo. The residue was recrystallized from isopropanol (200mL) to give the title compound as a pale yellow crystalline solid (13.3g, 45%).
MS(ES+)187.1(M+1)+。1H NMR(400MHz，CDCl3)9.81(br.s.，1H)7.39(d，J＝7.81Hz，1H)5.94(d，J＝8.00Hz，1H)4.39(dd，J＝5.27，4.49Hz，2H)3.73(dd，J＝5.07，4.29Hz，2H)3.36(s，3H)。
Preparation example 15
6-iodo-1- (2-methoxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one
N-butyllithium (2N in hexanes, 30.0mL, 60.0mmol) was added dropwise to a stirring solution of diisopropylamine (8.3mL, 59.10mmol) in tetrahydrofuran (50mL) at-78 deg.C under argon. The reaction mixture was slowly warmed to-20 ℃ and then cooled to-78 ℃. A solution of 1- (2-methoxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one (5.0g, 26.85mmol) in tetrahydrofuran (50mL) was added dropwise at-78 ℃. The reaction mixture was slowly warmed to-10 ℃ over 1h and then cooled to-78 ℃. A solution of iodine (15.0g, 59.07mmol) in tetrahydrofuran (50mL) was added at-78 deg.C and the reaction mixture was stirred at room temperature for 20 h. The reaction was diluted with saturated aqueous ammonium chloride (200mL) and the organic solvent was removed under reduced pressure. The aqueous residue was acidified to pH4 with 1N aqueous hydrochloric acid and extracted with dichloromethane (3X 300mL, 1X 200 mL). The combined organic layers were washed with 10% aqueous sodium thiosulfate (400mL), brine (300mL), dried over magnesium sulfate and concentrated in vacuo. The resulting residue was stirred in dichloromethane at room temperature and the solid was collected by filtration to give the title compound as a light brown solid (9.05g, 54%). The filtrate was concentrated and purified by flash chromatography (0-25% dichloromethane/ethyl acetate) to afford the title compound as a second creamy yellow solid (3.10g, 18%) (72% combined yield).
MS(ES+)313.0[M+1]+。1H NMR(400MHz，CDCl3)9.88(br.s.，1H)6.70(s，1H)4.88(br.s.，2H)3.78(t，J＝6.05Hz，2H)3.40(s，3H)。
Preparation example 16
6-iodo-1- (2-methoxyethyl) -2- (methylthio) pyrimidin-4 (1H) -one
Diisopropylethylamine (5.0mL, 28.83mmol) and iodomethane (9.0mL, 144.17mmol) were added to a stirring solution of 6-iodo-1- (2-methoxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one (9.00g, 28.83mmol) in acetonitrile (200 mL). The reaction mixture was stirred at room temperature for 18h and concentrated in vacuo. The residue was partitioned between dichloromethane (200mL) and 1N aqueous hydrochloric acid (100 mL). The layers were separated and the organic layer was washed with brine (100mL), dried over magnesium sulfate and concentrated in vacuo. The resulting residue was purified by hot milling with dichloromethane/heptane to give the title compound as a cream yellow solid (4.05g, 43%).
MS(ES+)327.0[M+1]+。1H NMR(500MHz，CDCl3)6.77(s，1H)4.42(t，J＝6.34Hz，2H)3.69(t，J＝6.34Hz，2H)3.40(s，3H)2.58(s，3H)。
Suzuki route section
Preparation example 17
6- (2, 5-Dimethoxyphenyl) -1- (2-methoxyethyl) -2- (methylthio) pyrimidin-4 (1H) -one
An aqueous solution (0.7mL) of degassed 1, 4-dioxane (2mL) followed by degassed sodium carbonate (65mg, 0.61mmol) was added to a mixture of 6-iodo-1- (2-methoxyethyl) -2- (methylthio) pyrimidin-4 (1H) -one (100mg, 0.31mmol), (2, 5-dimethoxyphenyl) boronic acid (0.37mmol, 1.2 equivalents) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (II) (14mg, 0.017mmol, 0.05 equivalents). The reaction mixture was subjected to microwave irradiation at 120 ℃ for 30min and the crude reaction mixture was used directly in the next step.
Example 9
6- (2, 5-Dimethoxyphenyl) -1- (2-methoxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one
Ammonium sulfide (1mL, 14.63mmol) and pyridine (1mL, 12.41mmol) were added to the crude reaction mixture (0.31mmol theoretical yield) from the previous Suzuki coupling reaction, and the mixture was subjected to microwave irradiation at 75 ℃ for 30 min. The reaction mixture was cooled to room temperature, taken up in dichloromethane (10mL) and water (10mL) and then basified with 2N sodium hydroxide. The layers were separated and the aqueous layer was extracted with dichloromethane (2X 10 mL). The aqueous layer was then acidified to pH6 with 2N aqueous hydrochloric acid and extracted with ethyl acetate (3X 10 mL). The combined organic extracts were dried over magnesium sulfate and concentrated in vacuo. The crude reaction mixture was purified by flash chromatography to afford the desired product as a solid (38mg, 38% over two steps).
MS(ES+)323.1[M+1]+。1H NMR(400MHz，CDCl3)10.14(br.s.，1H)，7.01(dd，J＝8.90，3.10Hz，1H)，6.89(d，J＝9.16Hz，1H)，6.80(d，J＝3.21Hz，1H)，5.84(d，J＝1.83Hz，1H)，4.70(dt，J＝13.74，4.35Hz，1H)，3.83-3.92(m，1H)，3.78-3.82(m，6H)，3.73-3.79(m，1H)，3.44(ddd，J＝9.96，5.84，3.89Hz，1H)，3.16(s，3H)。
Section of Negishi route
Preparation example 18
1- (2-methoxyethyl) -2- (methylsulfanyl) -6- (pyridin-2-yl) pyrimidin-4 (1H) -one
N-butyllithium (2.0M, 0.32mL, 0.64mmol) was slowly added to a solution of 2-bromopyridine (0.058mL, 0.61mmol) in anhydrous tetrahydrofuran (2mL) at-78 ℃. After 30min, anhydrous zinc chloride (92mg, 0.67mmol) was added and the reaction mixture was stirred for another 30min while warming to room temperature. 6-iodo-1- (2-methoxyethyl) -2- (methylthio) pyrimidin-4 (1H) -one (200mg, 0.61mmol), followed by tris (dibenzylideneacetone) dipalladium (0) (27mg, 0.03mmol), 2-dicyclohexylphosphino-2 ',6' -dimethoxybiphenyl (26mg, 0.06mmol), and N, N-dimethylformamide (2mL) were added to the reaction mixture, which was then heated to 80 ℃. After stirring overnight, the product was extracted with ethyl acetate (3X 10mL) and washed with water (3X 10 mL). The aqueous layer was then acidified to pH4 with 2M hydrochloric acid and the product was extracted with dichloromethane (3X 10mL) and dried over magnesium sulfate. The solvent was removed in vacuo to give a mixture of 1- (2-methoxyethyl) -2- (methylthio) -6- (pyridin-2-yl) pyrimidin-4 (1H) -one (37%) and 1- (2-methoxyethyl) -2- (methylthio) pyrimidin-4 (1H) -one (32%) as an orange oil (100 mg).
Example 10
1- (2-methoxyethyl) -6- (pyridin-2-yl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one
Crude 1- (2-methoxyethyl) -2- (methylthio) -6- (pyridin-2-yl) pyrimidin-4 (1H) -one (100mg, 0.36mmol), ammonium sulfide solution (0.2mL, 0.64mmol) and pyridine (0.2mL) were stirred in dioxane (2mL) at 70 ℃ for 4H. The reaction mixture was diluted with water (10mL), basified with 2M sodium hydroxide, and washed with dichloromethane (3X 10 mL). The aqueous layer was acidified to pH6 with 2M hydrochloric acid and the product was extracted with ethyl acetate (3X 10 mL). The combined organic layers were washed with water (3 × 10mL), brine (3 × 10mL), dried over magnesium sulfate, and concentrated in vacuo. The crude material was purified by mass directed autopurification using acidic methods to afford the product as a brown solid (3mg, 3%).
MS(ES+)264.07[M+H]+。1H NMR(400MHz，CDCl3)8.76(brs，1H)，7.92(br s，1H)，7.48-7.54(m，2H)，7.32(br d，1H)，5.95(br d，1H)，4.65(brs，2H)，3.64(brs，2H)。
The following examples of table 2 are prepared from the corresponding carboxylic acids to give the intermediate β -keto-esters following the preparation of the carboxylic acid route section described above, followed by other methods described above in the i.
TABLE 2 examples from carboxylic acid routes
The following examples of table 3 are prepared from the corresponding methyl ketones to give the intermediate β -keto-esters, following the preparation of the methyl ketone route section described above, and thereafter using other methods described in the i.
TABLE 3 examples from the methyl ketone route
The following examples of table 4 are prepared according to the aryl halide route section preparations described above from the corresponding aryl halides to give the intermediate β -keto-esters, which are then prepared using the methods described in the i.
TABLE 4 examples from aryl halide routes
The following examples of Table 5 were prepared from 6-iodo-1- (2-methoxyethyl) -2- (methylthio) pyrimidin-4 (1H) -one and the appropriate aryl borate following the preparative examples and procedures described above in the Suzuki route section and standard methods and techniques known to those skilled in the art.
TABLE 5 examples from the Suzuki route
I. Supplementation of the beta-ketoester route
E. Part of the ester route
Preparation example 19
3- (2- (2-Hydroxyethoxy) phenyl) -3-oxopropanoic acid tert-butyl ester
Freshly prepared lithium diisopropylamide (37mL of 1.85M in tetrahydrofuran) was added dropwise over a period of 15min to a solution of tert-butyl acetate (7.96g, 68.5mmol) in anhydrous tetrahydrofuran (100mL) at-78 deg.C and the mixture was stirred for 30 min. 2H-benzo [ e ] [1,4] dioxepin-5(3H) -one (2H-benzole [ e ] [1,4] dioxepin-5(3H) -one) (10.2g, 62.3mmol) was added dropwise as a solution in tetrahydrofuran (50mL) at-78 deg.C and stirring continued for an additional 30 min. Saturated sodium bicarbonate solution was added and the mixture was extracted with ethyl acetate. The organic layer was dried (sodium sulfate) and concentrated under reduced pressure to give the title compound as a yellow oil (6.0g, 77.9%) which was used directly in the next step without further purification.
MS(ES+)303.2[M+Na]+.1H NMR(500MHz，CDCl3)7.85(dd，J＝7.8，1.8Hz，1H)7.50(ddd，J＝8.5，7.1，1.8Hz，1H)7.05(td，J＝7.5，1.0Hz，1H)6.97(d，J＝8.4Hz，1H)4.16-4.20(m，2H)4.01(d，J＝4.3Hz，2H)3.90(s，2H)2.79(br.s，1H)1.33(s，9H)。
Preparation example 20
3- (2- (2-Hydroxyethoxy) phenyl) -3-oxopropanoic acid ethyl ester
A solution of tert-butyl 3- (2- (2-hydroxyethoxy) phenyl) -3-oxopropanoate (2.0g, 7.14mmol) in ethanol (20mL) was heated in a microwave reactor at 120 ℃ for 90 min. The mixture was cooled to room temperature and the solvent was concentrated under reduced pressure to give a yellow oil. The crude product was purified by flash chromatography (30-40% ethyl acetate: petroleum ether) to afford the title compound as a yellow solid.
Preparation example 21
(Z) -3- ((2-amino-2-oxoethyl) amino) -3- (2- (2-hydroxyethoxy) phenyl) acrylic acid ethyl ester
Triethylamine (3.21g, 31.7mmol) was added to a solution of ethyl 3- (2- (2-hydroxyethoxy) phenyl) -3-oxopropanoate (2g, 7.94mmol) and glycinamide hydrochloride (3.5g, 31.7mmol) in methanol (20mL) at room temperature. The mixture was stirred at 40 ℃ for 20 min. Acetic acid (1.9g, 31.7mmol) was added and the mixture was stirred at 80 ℃ for 18 h. The reaction mixture was cooled to room temperature and saturated sodium bicarbonate (200mL) was added. The organic layer was separated, dried (sodium sulfate) and concentrated to give a yellow solid. The solid was washed with ethyl acetate (20mL) and the residue was dried under reduced pressure to give the title compound as a white solid. This material was used directly without further purification.
Example 349
2- (6- (2- (2-hydroxyethoxy) phenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide
Trimethylsilyl isothiocyanate (1.7g, 12.9mmol) was added to a solution of ethyl (Z) -3- ((2-amino-2-oxoethyl) amino) -3- (2- (2-hydroxyethoxy) phenyl) acrylate (1.0g, 3.25mmol) in tetrahydrofuran (15mL) and the mixture was stirred at 80 ℃ for 18 h. The reaction mixture was cooled to room temperature, poured into an aqueous flask, and extracted with dichloromethane (3X 100 mL). The combined organic layers were dried and concentrated under reduced pressure to give a yellow solid, which was purified by flash chromatography (2-5% methanol: dichloromethane) to give the title compound (330mg, 31.7%) as a yellow solid.
MS(ES+)343.9[M+Na]。1H NMR(DMSO-d6)d：12.77(s，1H)，7.44-7.53(m，1H)，7.31(s，1H)，7.14-7.22(m，2H)，7.02(t，J＝7.5Hz，1H)，6.98(br.s.，1H)，5.74-5.82(m，1H)，5.34(br.s.，1H)，4.84(br.s.，1H)，4.00-4.16(m，2H)，3.93(br.s.，1H)，3.65(q，J＝4.4Hz，2H)。
Amide coupling scheme moieties
Preparation example 22
(Z) -3- (2, 4-Dimethoxyphenyl) -3- ((2-ethoxy-2-oxoethyl) amino) acrylic acid methyl ester
Glycine methyl ester hydrochloride (10.5g, 83.9mmol), then acetic acid (1.20mL, 21mmol) and triethylamine (8.5g, 83.9mmol) were added to a solution of methyl 3- (2, 4-dimethoxyphenyl) -3-oxopropanoate (5.0g, 21mmol) in ethanol (30mL) and the reaction mixture was heated at 100 ℃ for 18 h. After cooling to room temperature, the residue was partitioned between ethyl acetate and saturated aqueous ammonium chloride solution. The organic layer was washed with brine, dried over sodium sulfate and concentrated in vacuo. The crude product was dissolved in dichloromethane (10mL), filtered through a plug of silica gel (eluting with 15-35% ethyl acetate in heptane) and dried in vacuo to afford the title compound as a yellow solid (4.7g, 69%). This material was used in the next step without further purification.
MS(ES+)324.3[M+1]+。1H NMR(500MHz，CDCl3)8.95(br.s.，1H)7.14(d，J＝10.57Hz，1H)6.49(dd，J＝8.28，2.07Hz，1H)6.46(d，J＝2.07，1H)4.60(s，1H)4.16(q，J＝7.80Hz，2H)3.83(s，3H)3.80(s，3H)，3.69(s，3H)，1.24(t，J＝7.80Hz，3H)。
Preparation example 23
2- (6- (2, 4-Dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetic acid ethyl ester
(trimethylsilyl) isothiocyanate (12.9mL, 90.8mmol) was added to a solution of methyl (Z) -3- (2, 4-dimethoxyphenyl) -3- ((2-ethoxy-2-oxoethyl) amino) acrylate (4.68g, 15.1mmol) in 2-methyltetrahydrofuran (38 mL). The resulting solution was purged 3 times with nitrogen and the mixture was heated at 110 ℃ for 18 h. The mixture was cooled to room temperature and the solvent was removed under reduced pressure to give a red solid. This residue was suspended in a 3:1 heptane/ethyl acetate mixture (200mL) and stirred at room temperature for 1 h. The solid was filtered, triturated with dichloromethane (100mL), concentrated under reduced pressure, and dried in vacuo to afford the title compound as a pink solid (4.42g, 87%). This material was used in the next step without further purification.
MS(ES+)351.5[M+1]+。1H NMR(500MHz，CDCl3)9.91(br s，1H)7.13(d，J＝6.12Hz，1H)6.54(s，1H)6.51(d，J＝6.12Hz，1H)5.86(s，1H)5.44-5.40(m，1H)4.25-4.20(m，1H)4.16-4.06(m，2H)，3.86(s，3H)3.83(s，3H)，1.20(t，J＝6.12Hz，3H)。
Preparation example 24
2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetic acid
6N aqueous sodium hydroxide (16.9mL) was added to a solution of ethyl 2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetate (6.8g, 20.3mmol) in methanol (34mL) and the solution was stirred at 35 ℃ for 3H. The mixture was concentrated under reduced pressure and water (100mL) was added. The aqueous layer was washed with ethyl acetate (2X 200mL) and acidified to pH about 2 with concentrated HCl. The resulting aqueous acidic solution was extracted with ethyl acetate (3 × 200mL), and the combined organic layers were dried over sodium sulfate and concentrated under reduced pressure to give 6.53g (99%) of the title compound as a white solid.
MS(ES+)323.2[M+1]+。1H NMR(500MHz，CD3OD)7.16(d，J＝8.86Hz，1H)6.67(s，1H)6.64(d，J＝8.86Hz，1H)5.79(s，1H)5.52-5.40(m，1H)4.34-4.19(m，1H)3.87(s，3H)3.86(s，3H)。
Preparation example 25
(2- (2- (6- (2, 4-Dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetylamino) ethyl) carbamic acid tert-butyl ester
Tert-butyl (2-aminoethyl) carbamate (40g, 250mmol) and pyridine (30mL) were added to a solution of 2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetic acid (40g, 124mmol) in N, N-dimethylformamide (300mL), and the mixture was stirred at room temperature for 15 min. The solution was cooled to 0 ℃ and then purged with nitrogen 3 times. After 10min, a solution of 50% propanephosphonic anhydride (T3P) in N, N-dimethylformamide (109mL) was added dropwise at 0 ℃ with continued stirring for 1h, after which the water/ice bath was removed and stirring continued for 4 h. The reaction solution was slowly poured into a stirring aqueous hydrochloric acid solution (2500mL, 0.5M), and the suspension was stirred at room temperature for 1 h. The solid formed was filtered and the filter cake was washed with 0.5M hydrochloric acid solution (500mL) followed by water (500 mL). The solid was dried in a vacuum oven at 50 ℃ for 20h to give 54.6g of a pale beige powder. This solid was suspended in ethyl acetate (500mL), heated to 70 ℃ under a stream of nitrogen for 1h with stirring, and then at room temperature for 18 h. The suspension was cooled to 0 ℃ and the solid was filtered, the filter cake was washed with cold (0 ℃) ethyl acetate (100mL) and dried in a vacuum oven at 50 ℃ for 9h to give 49.0g of an off-white solid. The solid was suspended in acetonitrile (300mL) and stirred at 70 ℃ under a stream of nitrogen for 18 h. The mixture was cooled to 0 ℃ and the resulting solid was filtered, washed with cold acetonitrile (50mL) and dried in a vacuum oven at 50 ℃ for 8h to give 46.5g of an off-white solid. This solid was suspended in ethyl acetate (350mL) and heated to 70 ℃ under a stream of nitrogen for 1h with stirring and then at room temperature for 18 h. The suspension was cooled to 0 deg.C and the solid was filtered, the filter cake was washed with cold (0 deg.C) ethyl acetate (50mL) and dried in a vacuum oven at 50 deg.C for 9h to give the title compound as an off-white powder (45.4g, 78.8%).
MS(ES+)465.3[M+1]+。1H NMR(500MHz，CD3OD)8.99(br.s.，1H)7.16(d，J＝7.65Hz，1H)6.65(s，1H)6.62(d，J＝7.65Hz，1H)5.78(s，1H)5.51-5.41(m，1H)4.22-4.14(m，1H)3.87(s，3H)3.85(s，3H)3.19-3.11(m，2H)3.06-3.00(m，2H)1.42(s，9H)。
Example 241
N- (2-aminoethyl) -2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide hydrochloride
Acetyl chloride (1.55mL) was added dropwise over 5min to cold (0 ℃ C.) ethanol (21.5mL) under nitrogen, and the reaction mixture was heated at 50 ℃ for 30 min. The reaction mixture was cooled to room temperature and tert-butyl (2- (2- (6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetylamino) ethyl) carbamate (1.0g, 2.15mmol) was added, followed by heating to 50 ℃ for 1H. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was suspended in ethanol (10mL), heated to 75 ℃ for 20min, ethyl acetate (20mL) was added and heating continued for another 20 min. The mixture was slowly cooled to room temperature during 18h with stirring. The resulting precipitate was filtered and dried in a vacuum oven at 70 ℃ for 20h to give the title compound as a white solid (751mg, 87%).
MS(ES+)365.2[M+1]+。1H NMR(500MHz，DMSO-D6)12.81(br.s.，1H)8.26(br.s.，1H)8.01(br.s.，2H)7.08(d，J＝7.91Hz，1H)6.70(s，1H)6.62(d，J＝7.91Hz，1H)5.78(s，1H)5.41-5.35(m，1H)4.07-4.02(m，1H)3.84(s，3H)3.83(s，3H)3.20-3.16(m，2H)2.74-2.64(m，2H)。
Guanidine route section
Example 350
[ amino {3- [6- (5-chloro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] propyl } amino ] methylene ] carbamic acid methyl ester
N, N-diisopropylethylamine (0.024mL, 0.14mmol) was added to a solution of 1- (3-aminopropyl) -6- (5-chloro-2-methoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one (prepared in analogy to example 6; 50mg, 0.14mmol) and methyl [ amino (1H-pyrazol-1-yl) methylene ] carbamate (28mg, 0.16mmol) in N, N-dimethylformamide (0.46mL) at room temperature, and the mixture was stirred for 72H. The solvent was removed under reduced pressure, and the residue was dissolved in dimethylsulfoxide (0.9mL) and purified using mass-induced autopurification to give the title compound (4.9 mg).
MS(ES+)425.9[M+H]+Retention time 1.54min, method XBridge C185um, 4.6 × 50mm, linear to 5% water/95% acetonitrile at 95% water/5% acetonitrile over 4.0min, held at 5% water/95% acetonitrile to 5.0min (0.03% NH)4OH). Flow rate: 2 mL/min.
Preparation example 26
3, 3-Difluoroazetidine-1-carbonitrile
A suspension of 3, 3-difluoroazetidine hydrochloride (600mg, 4.63mmol) in dichloromethane (15.4mL) was treated with triethylamine (1.48 mL). The reaction mixture was cooled to 0 ℃ and treated with cyanogen bromide (3M in dichloromethane, 2.01mL, 6.02mmol) and the reaction mixture was stirred at 0 ℃ for 2 h. The reaction mixture was diluted with water (10mL) and saturated sodium bicarbonate (5mL) and extracted with ethyl acetate (3X 50mL) and dichloromethane (2X 50 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was dissolved in dichloromethane (30mL) and washed with saturated aqueous ammonium chloride (2X 15 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo to afford the title compound as a light brown solid (490mg, 89%).
1H NMR(500MHz，CDCl3)4.52(t，4H)。
Preparation example 27
1- (1H-benzotriazol-1-yl) -1- (3, 3-difluoroazetidin-1-yl) methanimine
A mixture of 3, 3-difluoroazetidine-1-carbonitrile (135mg, 1.14mmol) and benzotriazole (136mg, 1.14mmol) in 1, 2-dichloroethane (0.2mL) was heated to 80 ℃ for 30min under nitrogen. The needle was inserted to accelerate the evaporation of the solvent and the mixture was heated at 80 ℃ for 45 min. The resulting solid was washed with diethyl ether (2X 3mL) and dried under reduced pressure to give the title compound as an off-white solid (160mg, 51%).
1H NMR(500MHz，DMSO-d6)8.35(d，1H，J＝8.4Hz)，8.15(d，1H，J＝8.4Hz)，7.92(s，1H)，7.67(t，1H，J＝8.4Hz)，7.52(t，1H，J＝8.4Hz)，4.64(t，4H，J＝12.8Hz)。
Example 351
3, 3-difluoro-N- {3- [6- (5-fluoro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] propyl } azetidine-1-carboxamidine
N, N-diisopropylethylamine (0.061mL, 0.35mmol) was added to a solution of 1- (3-aminopropyl) -6- (5-fluoro-2-methoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one (prepared in analogy to example 6; 50mg, 0.14mmol) and 1- (1H-benzotriazol-1-yl) -1- (3, 3-difluoroazetidin-1-yl) methanimine (47.5mg, 0.174mmol) in N, N-dimethylformamide (0.46mL) under nitrogen and heated at 60 ℃ for 1H under nitrogen. The reaction mixture was cooled to room temperature and treated with 4N hydrochloric acid in dioxane (0.25 mL). The mixture was stirred at room temperature for 10min, then concentrated in vacuo and azeotroped with heptane (3X 10 mL). The residue was dissolved in water (1mL) and purified using medium pressure reverse phase (C18) chromatography (100:0 to 70:30 water/acetonitrile) to give the title compound as a white solid (22mg, 33%).
MS(ES+)428.2[M+H]+。1H NMR(500MHz，CD3OD)7.36(ddd，1H，J＝9.1，8.2，3.2Hz)，7.21-7.24(m，2H)，5.85(s，1H)，4.6(br s，1H)，4.45(td，4H，J＝11.4，4.7Hz)，3.92(s，3H)，3.80(br s，1H)，3.12(td，2H，J＝6.0，2.4Hz)，2.00-2.05(m，1H)，1.72-1.82(m，1H)。
The following examples of table 6 (supplement to table 2) were prepared from the corresponding carboxylic acids to give the intermediate β -keto-esters following the preparation of the carboxylic acid route section as described above, followed by other methods described in the i.
TABLE 6 examples from carboxylic acid routes
The following examples of table 7 (supplement to table 3) were prepared from the corresponding methyl ketones to give intermediate β -keto-esters following the preparation of the methyl ketone route section as described above, followed by other methods described in the i.
TABLE 7 examples from the methyl ketone route
The following examples of table 8 (supplement to table 4) were prepared according to the aryl halide route section preparations described above from the corresponding aryl halide to give the intermediate β -keto-ester, which was then prepared using the methods described in the i.
TABLE 8 examples from aryl halide routes
The following examples of table 9 are prepared according to the ester route section preparation described above from the corresponding aryl ester or lactone to give the intermediate β -keto-ester, which is then prepared using the methods described in the i.
TABLE 9 examples from the ester route
The following examples of table 10 are prepared from the corresponding thiouracil carboxylic acids according to the preparation of the iii amide coupling route section described above, and by using the methods described in the i.β -ketoester route section and standard methods and techniques known to those skilled in the art.
TABLE 10 examples from the amide coupling route
The following examples of table 11 were prepared from the corresponding thiouracils according to the iv. guanidine route section preparation described above, and by using the methods described in the i. beta. -ketoester route section and standard methods and techniques known to those skilled in the art.
TABLE 11 examples from the guanidine route
All publications, including but not limited to approved patents, patent applications, and journal articles, cited in this application are each incorporated by reference herein in their entirety.
While the present invention has been described in terms of the embodiments disclosed above, it will be readily apparent to those skilled in the art that the specific embodiments described in detail are merely illustrative of the invention. It will be understood that various modifications may be made without departing from the spirit of the invention.
Claims (25)
1. A compound having the formula I
Or a pharmaceutically acceptable salt thereof
Wherein
R1Is phenyl, naphthyl, furyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, quinolyl, isoquinolyl, pyrazolyl, pyrrolyl, indolyl, benzo [ b ]]Thienyl, benzothiazoleRadical, benzo [ b]Furyl or thienyl; and wherein said R1Independently by cyano group, (C)1-C4) Alkyl, (C)1-C4) Alkoxy, hydroxy (C)2-C4) Alkoxy, trifluoro (C)1-C4) Alkyl, trifluoro (C)1-C4) Alkoxy or halo mono-, di-or tri-substituted; and is
R2Is (C)1-C4) Alkoxy (C)1-C4) Alkyl, carboxyl (C)1-C4) Alkyl, mono-or di-hydroxy (C)2-C6) Alkyl, amino (C)2-C4) Alkyl, diamino methylene amino (C)2-C4) Alkyl, mono-N-or di-N, N (C)1-C4) Alkylamino radical (C)2-C4) Alkyl, (C)1-C4) Alkylcarbonyloxy (C)1-C4) Alkyl, (C)1-C4) Alkoxycarbonyl (C)1-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, carbamoylamino (C)2-C4) Alkyl, mono-N-or di-N, N (C)1-C4) Alkylcarbamoyl (C)1-C4) Alkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, (C)1-C4) Alkoxycarbonylamino (C)2-C4) Alkyl, (C)1-C4) Alkylsulfonylamino (C)2-C4) Alkyl, (C)1-C4) Alkylaminosulfonyl (C)1-C4) Alkyl, aminosulfonyl (C)1-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl or (C)1-C4) Alkylthio alkyl (C)1-C4)；
Provided that R is1Is not phenyl, and R2Is not (C)1-C6) An alkyl group.
2. The compound of claim 1, wherein
R1Is phenyl, naphthyl, pyridyl, quinolyl, isoquinolyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, furyl, pyrrolyl, indolyl, benzo [ b ]]Thienyl, benzothiazolyl, benzo [ b ]]Furyl or thienyl; wherein said R1Independently quilt (C)1-C4) Alkyl, (C)1-C4) Alkoxy, hydroxy (C)2-C4) Alkoxy, cyano, trifluoromethyl, trifluoromethoxy or halo mono-, di-or tri-substituted; and is
R2Is (C)1-C4) Alkoxy (C)1-C4) Alkyl, carboxyl (C)1-C4) Alkyl, mono-or di-hydroxy (C)2-C6) Alkyl, amino (C)2-C4) Alkyl, diamino methylene amino (C)2-C4) Alkyl, mono-N-or di-N, N (C)1-C4) Alkylamino radical (C)2-C4) Alkyl, (C)1-C4) Alkylcarbonyloxy (C)1-C4) Alkyl, (C)1-C4) Alkoxycarbonyl (C)1-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, carbamoylamino (C)2-C4) Alkyl, mono-N-or di-N, N (C)1-C4) Alkylcarbamoyl (C)1-C4) Alkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, (C)1-C4) Alkoxycarbonylamino (C)2-C4) Alkyl, (C)1-C4) Alkylsulfonylamino (C)2-C4) Alkyl, (C)1-C4) Alkylaminosulfonyl (C)1-C4) Alkyl, aminosulfonyl (C)1-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl or (C)1-C4) Alkylthio alkyl (C)1-C4)。
3. The compound of claim 2, wherein
R1Is phenyl and said R1Independently mono-, di-or tri-substituted with hydroxyethoxy, methyl, methoxy, fluoro or chloro; and is
R2Is diaminomethyleneamino (C)2-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, hydroxy (C)2-C4) Alkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl or amino (C)2-C4) An alkyl group.
4. The compound of claim 1, wherein
R1Is phenyl, naphthyl, pyridyl, quinolyl, isoquinolyl, pyrazolyl, pyrimidinyl, pyridazinyl, pyrazinyl, furyl, pyrrolyl, indolyl, benzo [ b ]]Thienyl, benzothiazolyl, benzo [ b ]]Furyl or thienyl; wherein said R1Independently quilt (C)1-C4) Alkyl, (C)1-C4) Alkoxy, hydroxy (C)2-C4) Alkoxy, cyano, trifluoromethyl, trifluoromethoxy or halo mono-, di-or tri-substituted; and is
R2Is triazolyl (C)1-C4) Alkyl, pyridyl (C)1-C4) Alkyl, pyrazinyl (C)1-C4) Alkyl, pyridazinyl (C)1-C4) Alkyl, pyrimidinyl (C)1-C4) Alkyl, imidazolyl (C)1-C4) Alkyl or pyrrolidinyl (C)1-C4) Alkyl radical, said R2The rings are optionally independently (C)1-C4) Alkyl, (C)1-C4) Alkoxy or halogen mono-, di-or tri-substituted.
5. The compound of claim 1, wherein R1Is phenyl and said R1Independently mono-, di-, tri-substituted with hydroxyethoxy, methyl, methoxy, fluoro or chloro.
6. The compound of claim 1, wherein R2Is hydroxy (C)2-C4) Alkyl, diamino methylene amino (C)2-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl or amino (C)2-C4) An alkyl group.
7. The compound of claim 1, wherein R2Independently by amino, carbamoyl, hydroxy, (C)1-C4) Alkoxy, amino (C)1-C4) Alkylcarbonylamino, amino (C)2-C4) Alkylcarbamoyl, (C)1-C4) Mono-or di-substituted (C) by alkylcarbonylamino or diaminomethyleneamino1-C4) An alkyl group.
8. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is
6- (2, 4-dimethoxyphenyl) -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
1- (2-aminoethyl) -6- (2, 4-dimethoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
2- [6- (2, 5-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
2- [6- (5-chloro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
1- (2-aminoethyl) -2-thioxo-6- (2,4, 5-trimethoxyphenyl) -2, 3-dihydropyrimidin-4 (1H) -one;
1- (3-aminopropyl) -6- (2-methoxy-5-methylphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
n- {2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] ethyl } glycinamide;
2- {3- [6- (2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] propyl } guanidine;
1- [ (2S) -3-amino-2-hydroxypropyl ] -6- (5-chloro-2-methoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
1- [ (2R) -3-amino-2-hydroxypropyl ] -6- (5-chloro-2-methoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] N- (2-aminoethyl) -acetamide; or
1- (2-aminoethyl) -6- [2- (2-hydroxyethoxy) phenyl ] -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one
Or a pharmaceutically acceptable salt thereof.
9. The compound 2- (6- (2, 5-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide or a pharmaceutically acceptable salt thereof.
10. The compound 2- (6- (5-chloro-2-methoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl) acetamide, or a pharmaceutically acceptable salt thereof.
11. A compound having the formula
12. A compound having the formula
13. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a cardiovascular condition in a mammal.
14. The use of claim 13, wherein the cardiovascular condition is heart failure, peripheral arterial disease, pulmonary hypertension, or vasculitis.
15. The use of claim 13, wherein the cardiovascular condition is congestive heart failure.
16. The use of claim 13, wherein the mammal has unstable angina or has experienced myocardial infarction.
17. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 or a pharmaceutically acceptable salt of said compound and a pharmaceutically acceptable carrier, excipient or diluent.
18. A pharmaceutical combination composition comprising: a therapeutically effective amount of a composition comprising:
a first compound that is a compound of claim 1 or a pharmaceutically acceptable salt of said compound;
a second compound that is an angiotensin converting enzyme inhibitor, an HMG-CoA reductase inhibitor, a non-steroidal anti-inflammatory drug, a factor Xa inhibitor, or warfarin; and
a pharmaceutically acceptable carrier, excipient or diluent.
19. The compound of claim 2, wherein
R1Is naphthyl, quinolyl, isoquinolyl, indolyl, benzo [ b ]]Thienyl, benzothiazolyl, benzo [ b ]]Furyl or thienyl and said R1Independently mono-, di-or tri-substituted with hydroxyethoxy, methyl, methoxy, fluoro or chloro; and is
R2Is diaminomethyleneamino (C)2-C4) Alkyl, carbamoyl (C)1-C4) Alkyl, hydroxy (C)2-C4) Alkyl, amino (C)2-C4) Alkylcarbamoyl (C)1-C4) Alkyl, (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)1-C4) Alkylcarbonylamino group (C)2-C4) Alkyl, amino (C)3-C4) Hydroxyalkyl or amino (C)2-C4) An alkyl group.
20. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is
2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
2- [6- (2-methoxy-5-methylphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
1- [ (2R) -2-aminopropyl ] -6- (2, 4-dimethoxyphenyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
2- [6- (3-methoxy-2-naphthyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide; or
2- [6- (1H-indol-4-yl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide.
21. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is
2- {6- [2- (2-hydroxyethoxy) -5-methoxyphenyl ] -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl } acetamide;
2- {6- [2- (2-hydroxyethoxy) -4-methoxyphenyl ] -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide;
6- [2- (2-hydroxyethoxy) -4-methoxyphenyl ] -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one;
6- [ 5-fluoro-2- (2-hydroxyethoxy) phenyl ] -1- (2-hydroxyethyl) -2-thioxo-2, 3-dihydropyrimidin-4 (1H) -one; or
2- {6- [2- (2-hydroxyethoxy) -4-methoxyphenyl ] -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl } acetamide.
22. The compound N- (2-aminoethyl) -2- [6- (2, 4-dimethoxyphenyl) -4-oxo-2-thioxo-3, 4-dihydropyrimidin-1 (2H) -yl ] acetamide or a pharmaceutically acceptable salt thereof.
23. A compound having the formula
24. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating cardiovascular events and conditions in a mammal, wherein the cardiovascular condition or event is heart failure, peripheral arterial disease, pulmonary hypertension, vasculitis, primary or secondary myocardial infarction, ischemia reperfusion injury, atrial fibrillation, or coronary artery bypass graft surgery.
25. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a condition in a mammal, wherein the condition is dialysis, delayed recovery of graft function, rejection of a transplanted organ, or nephropathy caused by a contrast agent.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201161558605P | 2011-11-11 | 2011-11-11 | |
| US61/558,605 | 2011-11-11 | ||
| PCT/IB2012/055949 WO2013068875A1 (en) | 2011-11-11 | 2012-10-28 | 2-thiopyrimidinones |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| HK1196603A1 HK1196603A1 (en) | 2014-12-19 |
| HK1196603B true HK1196603B (en) | 2017-08-04 |
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