Isoquinolinecarboxylic acid compounds, processes for their preparation and use

Abstract

The present application discloses isoquinoline carboxylic acid compounds, and preparation methods and applications thereof. The isoquinoline carboxylic acid compounds, or pharmaceutically acceptable salts, stereoisomers, tautomers, solvates or prodrugs thereof can be used as prolyl hydroxylase inhibitors.

Description

Technical Field

[0001] This invention relates to isoquinoline carboxylic acid compounds, pharmaceutical compositions containing them, and methods for treating conditions such as renal anemia mediated by prolyl hydroxylase activity. Background Technology

[0002] Anemia is a metabolic disorder of the blood, including a decrease in the number of red blood cells (RBCs) or hemoglobin in the bloodstream. Chronic kidney disease (CKD), cancer, chronic inflammatory lesions, nutritional and genetic defects, and inappropriate drug treatments can all lead to anemia. Chronic kidney disease, chemotherapy for cancer, and chronic inflammatory diseases result in reduced erythropoietin (EPO) production and a lack of stable iron. Treatment for anemia associated with CKD involves the use of EPO analogs or erythropoiesis-stimulating agents (ESAs). CKD often presents with iron deficiency requiring iron supplementation. Functional iron deficiency is a significant problem in most patients requiring erythropoiesis-stimulating agents (ESAS).

[0003] Homeostatic hypoxia-inducible factor (HIF) represents a novel approach to treating chronic kidney disease (CKD). Under normal conditions, HIF reduces oxidation via prolyl hydroxylase (PHD). Hypoxia is a major inhibitor of PHD, stabilizing HIF and promoting erythropoietin (EPO) synthesis. Prolyl hydroxylase (PHD) inhibitors can stabilize HIF. PHD inhibitors represent a new class of drug treatments for anemia in CKD. Many oral prolyl hydroxylase (PHD) inhibitors, such as roxadustat and vadadustat, are already marketed or in late-stage clinical trials.

[0004] Although existing technologies such as WO2004108681A1 disclose isoquinoline compounds for regulating HIF and / or EPO, their therapeutic effects remain unsatisfactory, significantly limiting their widespread clinical application. Therefore, there is still a need for new, effective compounds targeting HIF to treat conditions such as erythropoietin-related conditions, including anemia associated with diabetes, ulcers, kidney failure, cancer, infections, dialysis, surgery, and chemotherapy, and conditions involving localized ischemia and hypoxia, such as arterial occlusive disease, angina, intestinal infarction, pulmonary embolism, cerebral ischemia, and myocardial infarction. There is also a need for compounds that effectively prevent tissue damage caused by localized ischemia, which occurs due to conditions such as atherosclerosis, diabetes, and pulmonary diseases such as pulmonary embolism and similar conditions. In summary, this technology requires methods and compounds for regulating HIF and / or endogenous erythropoietin, and for treating and preventing HIF-related and EPO-related conditions, including those involving anemia, ischemia, and hypoxia. Summary of the Invention

[0005] Purpose of the invention: The technical problem to be solved by the present invention is to provide an isoquinoline carboxylic acid compound, or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof, or a combination thereof, in view of the shortcomings of the prior art.

[0006] A further technical problem to be solved by the present invention is to provide the use of the above-mentioned isoquinoline carboxylic acid compound, or its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate or prodrug and combinations thereof.

[0007] Another technical problem to be solved by the present invention is to provide a method for preparing the above-mentioned isoquinoline carboxylic acid compound.

[0008] To address the first technical problem mentioned above, this invention discloses an isoquinoline carboxylic acid compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or prodrug thereof:

[0009]

[0010] in,

[0011] R1 and R2 are each independently selected from the group consisting of the following groups: hydrogen, aryl, substituted aryl, heteroaryl, substituted heteroaryl, aryloxy, substituted aryloxy, heteroaryloxy, substituted heteroaryloxy;

[0012] R1' and R2' are each independently selected from hydrogen, alkyl, and halogen;

[0013] R8 is selected from methyl or cyclopropane as shown in formula (II);

[0014]

[0015] Among them, R3, R4, R5, R6, and R7 are each independently selected from halogen groups, hydrogen, deuterium, cyano, cyclohexyl, -SO2CH3, and -C. 1-10 Alkyl groups, or phenyl groups substituted at various positions.

[0016] Preferably, R1 and R2 are each independently selected from the group consisting of the following groups: hydrogen, phenyl, substituted phenyl, heteroaryl, substituted heteroaryl, phenoxy, substituted phenoxy, pyridoxy, pyrimidoxy, substituted pyridoxy, and substituted pyrimidoxy.

[0017] The above substitutions are selected from alkyl substitution, cycloalkyl substitution, alkoxy substitution, cyano substitution, halogroup substitution, and -SO2CH substitution.

[0018] Wherein, the alkyl group is C 1-10 C is further preferred. 1-4 More preferably, methyl, ethyl, propyl, or isopropyl.

[0019] The cycloalkyl group is cyclohexyl.

[0020] Wherein, the alkoxy group is C 1-10 The oxygen group is further preferably methoxy, ethoxy, propoxy, or isopropoxy.

[0021] The halogenated group is fluorine, chlorine, or bromine.

[0022] The number of substitutions is mono-substitution, di-substitution, tri-substitution, or complete substitution.

[0023] More preferably, R1 and R2 are each independently selected from the group consisting of the following radicals: 2,6-dimethylphenoxy, 3,4-difluorophenoxy, 3,5-difluorophenoxy, 3-chloro-4-fluorophenoxy, 3-methoxy-4-fluorophenoxy, 3-methoxy-5-fluorophenoxy, 4-chlorophenoxy, 4-fluorophenoxy, 4-methoxyphenoxy, 3-chlorophenoxy, 4-chlorophenoxy, and 3,4-dichlorophenoxy.

[0024] More preferably, the isoquinoline carboxylic acid compound represented by formula (I) has the following structural formula:

[0025]

[0026] More preferably, the isoquinoline carboxylic acid compound represented by formula (I) is selected from the compounds represented by formulas 2i, 2j, 3g, and 3h.

[0027] Most preferably, the isoquinoline carboxylic acid compound represented by formula (I) is selected from the compounds represented by formula 3g and formula 3h.

[0028] A composition containing an effective amount of an isoquinoline carboxylic acid compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate or prodrug thereof, and a pharmaceutically acceptable excipient is also within the scope of protection of this invention.

[0029] To address the second technical problem mentioned above, this invention discloses the use of the above-mentioned compound or its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, or the pharmaceutical composition thereof in the preparation of a medicament for treating, preventing, or pre-treating diseases at least partially mediated by hypoxia-inducible factor and / or erythropoietin.

[0030] The disease is selected from the group consisting of the following conditions: anemia, neurological conditions and / or injuries, peripheral vascular conditions, ulcers, burns and chronic wounds, pulmonary embolism, ischemia-reperfusion injury; further, the disease is renal anemia.

[0031] The present invention also discloses the use of the above-mentioned compound or its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate or the pharmaceutical composition thereof in the preparation of a medicament for inhibiting the activity of prolyl hydroxylase, including contacting the prolyl hydroxylase enzyme with the above-mentioned compound or its pharmaceutically acceptable salt, stereoisomer, tautomer or solvate.

[0032] To address the third technical problem mentioned above, this invention discloses a method for preparing the isoquinoline carboxylic acid compound represented by formula (I).

[0033] Wherein, R8 is selected from cyclopropane, R1 and R2 are each independently selected from hydrogen and phenoxy groups, and R1 and R2 are not both hydrogen, the preparation method of the compound includes the following steps:

[0034] Step 1: Using the 4-nitrophthalonitrile derivative shown in Formula 4a as a starting material, prepare the diaryl ether shown in Formula 4c via a nucleophilic substitution reaction;

[0035] Step 2: The diaryl ether shown in Formula 4c is prepared into the dicarboxylic acid shown in Formula 4d by cyano hydrolysis;

[0036] Step 3: The dicarboxylic acid shown in Formula 4d is cyclized with ammonia to obtain the cyclized carboxylic acid shown in Formula 4f;

[0037] Step 4: The cyclized carboxylic acid shown in Formula 4f is hydrolyzed to obtain the ester shown in Formula 4g;

[0038] Step 5: The ester shown in Formula 4g is subjected to Dickman condensation to give the ring-expanded products shown in Formulas 4h and 4i;

[0039] Step 6: The ring-expansion products shown in formulas 4h and 4i are subjected to bromination to obtain the bromoisoquinoline derivative esters shown in formulas 4j and 4k;

[0040] Step 7a: The bromoisoquinoline derivative esters shown in Formulas 4j and 4k are used to prepare the three-membered ring isoquinoline derivatives shown in Formulas 4l and 4m via a Suzuki coupling reaction.

[0041] Step 8a: The three-membered ring isoquinoline derivatives shown in formulas 4l and 4m are obtained by ester hydrolysis to the carboxylic acids shown in formulas 4n and 4o;

[0042] Step 9a: The carboxylic acids represented by formulas 4n and 4o are condensed with glycine methyl ester or its hydrochloride to obtain the amides represented by formulas 4p and 4q;

[0043] Steps 10a and 10b: The amides represented by formulas 4p and 4q are obtained by ester hydrolysis to the carboxylic acids represented by formulas 4r and 4s, respectively;

[0044]

[0045] R1' and R2' are each independently selected from hydrogen, alkyl, and halogen.

[0046] In step (1), the reaction is a reaction between a 4-nitrophthalonitrile derivative of formula 4a and a phenolic compound in a solvent.

[0047] The molar ratio of the 4-nitrophthalonitrile derivative to the phenolic compound is 1:0.5-1.5, preferably 1:1.

[0048] The solvents include, but are not limited to, dimethyl sulfoxide.

[0049] The concentration of the 4-nitrophthalonitrile derivative is 1-1.5 mmol / mL, preferably 1.325 mmol / mL.

[0050] The reaction raw materials also include potassium carbonate.

[0051] The molar ratio of the 4-nitrophthalonitrile derivative to potassium carbonate is 1:0.5-1.5, preferably 1:1.

[0052] The reaction temperature is room temperature - 80°C; preferably, the mixture is first stirred at room temperature and then heated to 60°C to react.

[0053] In step (2), the diaryl ether represented by formula 4c undergoes hydrolysis in a mixed solvent of water and methanol in the presence of sodium hydroxide.

[0054] The concentration of the diaryl ether represented by Formula 4c is 0.1-0.2 mmol / mL, preferably 0.156 mmol / mL.

[0055] The volume ratio of water to methanol is 1-2:1, preferably 1.5:1.

[0056] The molar ratio of the diaryl ether shown in Formula 4c to sodium hydroxide is 1:15-25, preferably 1:18-20.

[0057] The reaction temperature is 80-120℃, preferably 100℃.

[0058] In step (3), the molar ratio of the dicarboxylic acid shown in formula 4d to ammonia is 1:0.8-1.2, preferably 1:1-1.1.

[0059] The solvent for the reaction is glacial acetic acid.

[0060] The concentration of the dicarboxylic acid represented by formula 4d is 0.2-0.6 mmol / mL, preferably 0.3-0.4 mmol / mL.

[0061] The reaction temperature is 150-180℃, preferably 160℃.

[0062] In step (4), the cyclized carboxylic acid represented by formula 4f is hydrolyzed in methanol under the action of concentrated sulfuric acid.

[0063] The ratio of the cyclic carboxylic acid shown in Formula 4f to methanol is 0.08-0.2 mmol / mL, preferably 0.11 mmol / mL.

[0064] The concentrated sulfuric acid is sulfuric acid with a mass fraction of 98%.

[0065] The ratio of the cyclic carboxylic acid shown in Formula 4f to concentrated sulfuric acid is 60-70 mmol / mL, preferably 66 mmol / mL.

[0066] The reaction temperature is 50-90℃, preferably 70℃.

[0067] In step (5), the Dickman condensation reaction is a first reaction between metallic sodium and n-butanol under an inert gas, followed by the addition of the ester represented by formula 4g to the reaction solution for a second reaction.

[0068] The ratio of sodium metal to n-butanol is 0.15-0.2 mmol / mL, preferably 0.175 mmol / mL.

[0069] The temperature of the first reaction is 40-80℃, preferably 60℃.

[0070] The reaction time for the first reaction is 5-40 min, preferably 5-20 min, and more preferably 10 min.

[0071] The molar ratio of the metallic sodium to the ester represented by Formula 4g is 2-3:1, preferably 7:3.

[0072] The temperature of the second reaction is 80-110℃, preferably 95℃.

[0073] In step (6), the ring-expanded products shown in formulas 4h and 4i react with the brominating agent in a solvent.

[0074] The solvents include, but are not limited to, acetonitrile.

[0075] The concentration of the ring-expanded products shown in formulas 4h and 4i is 0.03-1 mmol / mL, preferably 0.06 mmol / mL.

[0076] The brominating reagents include, but are not limited to, phosphorus tribromooxyphosphate.

[0077] The molar ratio of the ring-expanding product to the brominating reagent shown in formulas 4h and 4i is 1:5-7, preferably 9:52.

[0078] The reaction temperature is 60-100℃, preferably 80℃.

[0079] In step (7a), the coupling reaction is achieved by a coupling agent in a mixed solvent of toluene and water using a bromoisoquinoline derivative ester and a boric acid conjugate of cyclopropane as shown in formulas 4j and 4k.

[0080] The cyclopropane borate compound is cyclopropylboronic acid.

[0081] The molar ratio of the bromoisoquinoline derivative ester and the boric acid conjugate of cyclopropane shown in formulas 4j and 4k is 0.8-0.9:1, preferably 0.88:1.

[0082] The coupling agent used is a metal Pd compound, preferably a 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloromethane complex (Pd(dppf)Cl2·CH2Cl2).

[0083] The molar ratio of the coupling agent used to the bromoisoquinoline derivative esters shown in formulas 4j and 4k is 0.01-1, preferably 0.01-0.2, and more preferably 0.05-0.1.

[0084] The volume ratio of toluene to water is 15-30:1, preferably 20:1.

[0085] The concentration of the bromoisoquinoline derivative esters shown in Formulas 4j and 4k is 0.03-0.05 mmol / mL, preferably 0.04-0.042 mmol / mL.

[0086] The reaction further includes cesium carbonate. Preferably, the molar ratio of the bromoisoquinoline derivative esters shown in Formulas 4j and 4k to cesium carbonate is 0.4-0.6:1, and more preferably 8.8:18.

[0087] The coupling reaction is carried out at a temperature of 50-120°C.

[0088] In step (8a), the solvent for hydrolysis is a mixed solution of methanol and water. Preferably, the volume ratio of methanol to water is 1.5-3:1, and more preferably 2:1.

[0089] The concentration of the three-membered cyclic isoquinoline derivative shown in Formulas 4l and 4m is 0.02-0.05 mmol / mL, preferably 0.03 mmol / mL.

[0090] The hydrolysis reaction further includes sodium hydroxide. Preferably, the molar ratio of the three-membered ring isoquinoline derivative shown in Formulas 4l and 4m to sodium hydroxide is 1:8-13, more preferably 1:10-11.

[0091] The reaction temperature is 50-90℃, preferably 70℃.

[0092] In step (9a), the condensation reaction is as follows: the carboxylic acid represented by formulas 4n and 4o reacts with dichloromethane and oxalyl chloride in a third reaction until no more bubbles are released. After removing the solvent, dichloromethane, glycine methyl ester or its hydrochloride, and diisopropylethylamine are added to carry out a fourth reaction.

[0093] In the third reaction, the concentration of the carboxylic acid represented by formulas 4n and 4o is 0.1-0.5 mmol / mL, preferably 0.325 mmol / mL.

[0094] In the third reaction, the molar ratio of the carboxylic acid represented by formulas 4n and 4o to oxaloyl chloride is 1:1.2-1.8, preferably 1:1.51.

[0095] The temperature of the third reaction is 0-30℃, preferably room temperature.

[0096] In the fourth reaction, the molar ratio of the carboxylic acid represented by formulas 4n and 4o to glycine methyl ester or its hydrochloride is 1:1.4-1.8, preferably 1:1.59.

[0097] In the fourth reaction, the ratio of dichloromethane, glycine methyl ester or its hydrochloride, and diisopropylethylamine is 1 mL: 0.4-0.7 mmol: 1-3 mmol, preferably 1 mL: 0.62 mmol: 2 mmol.

[0098] The temperature of the fourth reaction is room temperature.

[0099] In step (10a), the amide hydrolyzed to the form of formula 4p is reacted in a mixed solution of water and methanol in the presence of anhydrous lithium chloride.

[0100] The concentration of the amide represented by formula 4p is 0.01-0.03 mmol / mL, preferably 0.52 / 30 mmol / mL.

[0101] The volume ratio of water to methanol is 1:10-20, preferably 1:15.

[0102] The ratio of the amide shown in Formula 4p to anhydrous lithium chloride is 0.004-0.006 mmol / mg, preferably 0.0052 mmol / mg.

[0103] The reaction was carried out at room temperature.

[0104] Steps (10b) and (10a) only require replacing the amide shown in Formula 4p with the amide shown in Formula 4q.

[0105] Wherein, R8 is selected from methyl, R1 and R2 are each independently selected from hydrogen and phenoxy, and R1 and R2 are not both hydrogen, the method for preparing the compound includes the following steps:

[0106] Step 1: Using the 4-nitrophthalonitrile derivative shown in Formula 4a as a starting material, prepare the diaryl ether shown in Formula 4c via a nucleophilic substitution reaction;

[0107] Step 2: The diaryl ether shown in Formula 4c is prepared into the dicarboxylic acid shown in Formula 4d by cyano hydrolysis;

[0108] Step 3: The dicarboxylic acid shown in Formula 4d is cyclized with ammonia to obtain the cyclized carboxylic acid shown in Formula 4f;

[0109] Step 4: The cyclized carboxylic acid shown in Formula 4f is hydrolyzed to obtain the ester shown in Formula 4g;

[0110] Step 5: The ester shown in Formula 4g is subjected to Dickman condensation to give the ring-expanded products shown in Formulas 4h and 4i;

[0111] Step 6: The ring-expansion products shown in formulas 4h and 4i are subjected to bromination to obtain the bromoisoquinoline derivative esters shown in formulas 4j and 4k;

[0112] Step 7b: The bromoisoquinoline derivative esters shown in Formulas 4j and 4k are used to prepare the three-membered ring isoquinoline derivatives shown in Formulas 5l and 5m via a Suzuki coupling reaction;

[0113] Steps 8b and 8c: The three-membered ring isoquinoline derivatives shown in formulas 5l and 5m are obtained by ester hydrolysis to obtain the carboxylic acids shown in formulas 5n and 5o, respectively;

[0114] Steps 9b and 9c: The carboxylic acids represented by formulas 5n and 5o are respectively condensed with glycine methyl ester or its hydrochloride to obtain the amides represented by formulas 5p and 5q;

[0115] Steps 10c and 10d: The amides represented by formulas 5p and 5q are obtained by ester hydrolysis to the carboxylic acids represented by formulas 5r and 5s, respectively;

[0116]

[0117] R1' and R2' are each independently selected from hydrogen, alkyl, and halogen.

[0118] In step (7b), the coupling reaction is achieved by the bromoisoquinoline derivative esters, potassium carbonate, trimethylboroxane, and tetratriphenylphosphine palladium shown in formulas 4j and 4k via a coupling agent on a dioxane ring.

[0119] The concentration of the bromoisoquinoline derivative esters shown in Formulas 4j and 4k is 0.03-0.06 mmol / mL, preferably 0.044 mmol / mL.

[0120] The molar ratio of the bromoisoquinoline derivative ester, potassium carbonate, trimethylboroxane and tetratriphenylphosphine palladium shown in formulas 4j and 4k is 1:5-7:2-3:0.07-0.09, preferably 3.3:20.2:7:0.27.

[0121] The coupling reaction is carried out at a temperature of 50-120°C.

[0122] In step (8b), the solvent for hydrolysis is a mixture of methanol and water. Preferably, the volume ratio of methanol to water is 1.5-3:1, and more preferably 2.5:1.

[0123] The concentration of the three-membered ring isoquinoline derivative shown in Formula 5l is 0.01-0.04 mmol / mL, preferably 0.022 mmol / mL.

[0124] The hydrolysis reaction further includes sodium hydroxide. Preferably, the molar ratio of the three-membered ring isoquinoline derivative shown in Formula 51 to sodium hydroxide is 1:3-7, and more preferably 1:4.48.

[0125] The reaction temperature is 50-90℃, preferably 70℃.

[0126] In step (9b), the condensation reaction is as follows: the carboxylic acid represented by formula 5n reacts with dichloromethane and oxalyl chloride in the fifth reaction until no more bubbles are released. After removing the solvent, dichloromethane, glycine methyl ester or its hydrochloride, and diisopropylethylamine are added to carry out the sixth reaction.

[0127] In the fifth reaction, the concentration of the carboxylic acid represented by formula 5n is 0.01-0.05 mmol / mL, preferably 0.45 mmol / mL.

[0128] In the fifth reaction, the molar ratio of the carboxylic acid represented by formula 5n to oxaloyl chloride is 1:3-4, preferably 1:3.52.

[0129] The temperature of the fifth reaction is 0-30℃, preferably room temperature.

[0130] In the sixth reaction, the molar ratio of the carboxylic acid represented by formula 5n to glycine methyl ester or its hydrochloride is 1:5-15, preferably 1:10.22.

[0131] In the sixth reaction, the ratio of dichloromethane, glycine methyl ester or its hydrochloride, and diisopropylethylamine is 1 mL: 0.6-0.9 mmol: 1-3 mmol, preferably 15 mL: 6.85 mmol: 10.8 mmol.

[0132] The temperature of the sixth reaction is room temperature.

[0133] In step (10c), the amide hydrolyzed to the form of formula 5p is reacted in a mixed solution of water and methanol in the presence of anhydrous lithium chloride.

[0134] The concentration of the amide represented by formula 5p is 0.01-0.03 mmol / mL, preferably 0.0225 mmol / mL.

[0135] The volume ratio of water to methanol is 1:0.5-3, preferably 1:1.

[0136] The molar ratio of the amide shown in Formula 5p to anhydrous lithium chloride is 1:10-16, preferably 0.45:5.9.

[0137] The reaction was carried out at room temperature.

[0138] Steps (8d), (9d), and (10d) are similar to steps (8c), (9c), and (10c), except that the compounds shown in formulas 5l, 5n, and 5p are replaced with the compounds shown in formulas 5m, 5o, and 5q, respectively.

[0139] In the above process, the bromoisoquinoline derivatives prepared usually have isomer structures. Since the isomers have similar structures and polarities, the intermediates can directly enter the next step of the reaction without separation. Different isomers of the target product can be obtained by preparation.

[0140] The term "pharmaceutically acceptable salt" as used in this invention refers to a salt of the free acid or base of the compound represented by formula (I), which is non-toxic, biologically tolerable, or in other words, biologically suitable for administration to a subject. Generally, see G.S. Paulekuhn, et al., "Trends in Active Pharmaceutical Ingredient Salt Selection based Analysis of the Orange Book Database," J. Med. Chem., 2007, 50: 6665-72; S.M. Berge, et al., "Pharmaceutical Salts," J. Pharm Sci., 1977, 66: 1-19; and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahland Wermuth, Eds., Wiley-VC Handbook. VHCA, Zurich, 2002 (GS Paulekuhn et al., “Trends in Selecting Active Pharmaceutical Ingredient Salts Based on Analysis of the Orange Book Database,” *Journal of Pharmaceutical Chemistry*, 2007, Vol. 50, pp. 6665-6672; SMBerge et al., “Pharmaceutical Salts,” *Journal of Pharmaceutical Science*, 1977, Vol. 66, pp. 1-19; and *Pharmaceutical Salts: Properties, Selection and Applications*, edited by Stahl and Wermuth, Wiley-VCH and VHCA, Zurich, 2002). Examples of pharmaceutically usable salts are those that are pharmacologically effective and suitable for contact with patient tissues without undue toxicity, irritation, or allergic reactions. Compounds of formula (I) may have sufficiently acidic groups, sufficiently basic groups, or both types of functional groups, thereby reacting with a variety of inorganic or organic bases, as well as inorganic and organic acids, to form pharmaceutically usable salts.

[0141] Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, octanoates, acrylates, formates, isobutyrates, hexanoates, heptanoates, propynates, oxalates, malonates, succinates, octanoates, sebates, fumarates, maleates, butyn-1,4-dicitates, hexyn-1,6-dicitates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, alkylbenzoates, oxybenzoates, phthalates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, γ-hydroxybutyrate, glycolates, tartrates, methanesulfonates, propanesulfonates, zeolite-1-sulfonates, thiazoline-2-sulfonates, and mandelates.

[0142] The compound represented by formula (I) contains basic nitrogen, and the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, by treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, boric acid, phosphoric acid, etc., or by treating the free base with an organic acid such as acetic acid, phenylacetic acid, propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid, hydroxyethanesulfonic acid, succinic acid, valeric acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, oleic acid, palmitic acid. Lauric acid, arsenic acid (such as glucuronic acid or galacturonic acid), α-hydrocarbon acid (such as mandelic acid, citric acid or tartaric acid), amino acid (such as aspartic acid, glutaric acid or glutamic acid), aromatic acid (such as benzoic acid, 2-acetoxybenzoic acid, carboxylic acid or cinnamic acid), sulfonic acid (such as laurylsulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid), any compatible mixture of acids such as those given by way of example in this invention, and any other acid and mixture thereof that are considered equivalent or acceptable substitutes according to the ordinary skill level in this art.

[0143] The compounds of formula (I) also contain carboxylic acids, and the desired pharmaceutically acceptable salts can be prepared by any suitable method, for example by treating the free acid with an inorganic or organic base such as amines (primary, secondary, or tertiary amines), alkali metal hydroxides, alkaline earth metal hydroxides, any compatible mixtures of bases such as those given by way of example in this invention, and any other bases and mixtures thereof that are considered equivalents or acceptable substitutes according to the ordinary skill level of this art. Exemplary examples of suitable salts include organic salts derived from substances such as amino acids (e.g., N-methyl-D-glucosamine, lysine, choline, glycine, and arginine), ammonia, carbonates, bicarbonates, primary amines, secondary amines, tertiary amines, and cyclic amines (e.g., aminobutanetriol, benzylamine, pyrrolidine, piperidine, morpholine, and piperazine), and inorganic salts derived from substances such as sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.

[0144] The prodrugs described in this invention may include compounds having an amino acid residue or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues covalently linked to a carboxylic acid group of formula (I) via an amide bond or ester bond. Examples of amino acid residues include twenty naturally occurring amino acids typically identified by three-letter symbols, as well as 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, valine, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone.

[0145] Other types of prodrugs can be prepared by derivatizing a free carboxylic acid of formula (I) into an amide or alkyl ester. Examples of amides include those derived from ammonia, C... 1-6 Alkyl primary amines and di(C) 1-6 Alkyl) secondary amines. Secondary amines include 5- or 6-membered heterocyclic alkyl or heteroaryl ring moieties. Examples of acidic amines include those derived from ammonia. 1-3 Alkyl primary amines and di(C) l-2 Amides of alkylamines. Examples of esters in this invention include C. 1-7 Alkyl ester, C 5-7 Cycloalkyl esters, phenyl esters and (C 1-6Alkyl)phenyl esters. Preferably, the ester includes methyl esters. Prodrugs can also be prepared by derivatizing a free hydroxyl group with groups including hemisuccinate, phosphate, dimethylaminoacetic acid, and phosphoryloxymethoxycarbonyl groups, according to methods such as those described in Fleisher et al., Adv. Drug Delivery Rev. 1996, 19, 115-130 (Fleisher et al., Review of Advances in Drug Delivery, 1996, Vol. 19, pp. 115-130). Carbamate derivatives of hydroxyl and amino groups can also produce prodrugs. Carbonate derivatives, sulfonate esters, and sulfate esters of hydroxyl groups can also provide prodrugs. Alkyl derivatization into (acyloxy)methyl esters and (acyloxy)ethyl esters, wherein the acyl group may be an alkyl ester optionally substituted with one or more ether, amine, or acetic acid functional groups, or wherein the acyl group is an amino acid ester as described above, can also be used to produce prodrugs. This type of prodrug can be prepared as described in Robinson et al., J MedChem. 1996, 39(1), 10-18 (Robinson et al., Journal of Medicinal Chemistry, 1996, Vol. 39, No. 1, pp. 10-18). The free amine can also be derivatized into phthalamides, sulfonamides, or phosphorophthalamides. All of these prodrug moieties can be incorporated with functional groups including ethers, amines, and carboxylic acids.

[0146] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0147] This invention provides a novel compound as a prolyl hydroxylase inhibitor, its preparation method, and its uses, expanding the range of drugs for treating erythropoietin-related diseases, and achieving excellent inhibitory activity against prolyl hydroxylase, which can be used to prepare drugs that inhibit prolyl hydroxylase activity. Detailed Implementation

[0148] The following embodiments are used to further describe the present invention, but these embodiments are not intended to limit the scope of the present invention.

[0149] Example 1: Preparation of [(1-cyclopropyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (1r)

[0150]

[0151] Step 1: Preparation of 4-(3-chlorophenoxy)phthalonitrile (1c)

[0152]

[0153] 4-Nitrophthalonitrile (1a) (9.2 g, 53.0 mmol), 3-chlorophenol (1b) (6.8 g, 53.0 mmol), dimethyl sulfoxide (40 mL), and potassium carbonate (7.3 g, 53.0 mmol) were added sequentially to a three-necked flask. The mixture was stirred at room temperature for 2 hours, then heated to 60 °C for 4 hours. TLC showed almost no reactants remaining, at which point heating was stopped. The reaction mixture was cooled to room temperature and slowly poured into 300 mL of water, precipitating a brick-red solid. The solid was filtered, and the filter cake was dried to give 12.6 g of the red solid compound, with a yield of 93.1%.

[0154] Step 2: Preparation of 4-(3-chlorophenoxy)phthalic acid (1d)

[0155]

[0156] 4-(3-chlorophenoxy)phthalonitrile (1c) (10.0 g, 39.0 mmol), water (150 mL), methanol (100 mL), and sodium hydroxide (30.0 g, 750 mmol) were sequentially added to a 500 mL three-necked flask. The mixture was stirred for half an hour and then refluxed at 100 °C for 10 h. TLC analysis showed that the starting material had disappeared, at which point the reaction was stopped and the mixture was cooled to room temperature. The reaction solution was concentrated under reduced pressure to remove impurities, and the pH was adjusted to 3 with hydrochloric acid, resulting in the precipitation of a large amount of pale yellow solid. This solid was filtered, and the filter cake was dried to give 11.6 g of the compound as a white solid, with a yield of 100%.

[0157] Step 3: Preparation of 2-(5-m-chlorophenoxy-1,3-dioxo-2,3-dihydro-isoindolyl)acetic acid (1f)

[0158]

[0159] Compound 4-(3-chlorophenoxy)phthalic acid (1d) (11.0 g, 38.0 mmol), glycine (1e) (2.96 g, 39.4 mmol), and glacial acetic acid (110 mL) were sequentially added to a 250 mL three-necked flask. The mixture was heated to 160 °C and reacted for 5 hours. TLC monitoring showed that the starting material had disappeared, at which point the reaction was stopped and the mixture was cooled to room temperature. The reaction solution was concentrated under reduced pressure and slowly poured into 400 mL of water, precipitating a large amount of white solid. This solid was filtered, and the filter cake was dried to obtain 11.3 g of white solid powder, with a yield of 90.6%.

[0160] Step 4: Preparation of methyl 2-(5-m-chlorophenoxy-1,3-dioxo-2,3-dihydro-isoindolyl)acetate (1g)

[0161]

[0162] Compound 2-(5-m-chlorophenoxy-1,3-dioxo-2,3-dihydro-isoindolyl)acetic acid (1f) (11.0 g, 33.0 mmol) and methanol (300 mL) were added sequentially to a 500 mL single-necked flask, followed by the addition of 0.5 mL of concentrated sulfuric acid (98% by mass). The reaction was carried out at 70 °C for 7 h. TLC was used to detect the disappearance of the starting material, and the solid was precipitated by vacuum concentration. The solid was dissolved in 300 mL of ethyl acetate and poured into 200 mL of 1N sodium carbonate aqueous solution. The pH was adjusted to 8-9, and the mixture was separated. The organic phase was washed with 200 mL of saturated brine, dried over anhydrous sodium sulfate, decolorized with activated carbon, concentrated to dryness, slurried with petroleum ether, filtered, and dried to obtain 10.7 g of white powder, with a yield of 93.3%.

[0163] Step 5: Preparation of 1,4-dihydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester and 1,4-dihydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester (1h and 1i)

[0164]

[0165] Sodium metal (1.6 g, 70.0 mmol) was added to n-butanol (400 mL), and nitrogen was purged three times. The reaction was carried out at 60 °C for 10 minutes. Then, methyl 2-(5-m-chlorophenoxy-1,3-dioxo-2,3-dihydro-isoindolyl)acetate (1 g) (10.5 g, 30.0 mmol) was added to the reaction system, and the temperature was raised to 95 °C for 4 hours. TLC analysis showed that the starting material disappeared. The reaction solution was poured into hydrochloric acid aqueous solution, and the pH was adjusted to 6-7, resulting in the precipitation of a large amount of white solid. The solid was filtered, the filter cake was dried, and column chromatography yielded 7.8 g of white solid, with a yield of 66.2%. This white solid was a mixture of butyl 1,4-dihydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate and butyl 1,4-dihydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate.

[0166] Step Six: Preparation of 1-bromo-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester and 1-bromo-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester (1j and 1k)

[0167]

[0168] Phosphorus tribromooxy (30.0 g, 104 mmol), acetonitrile (300 mL), the mixture obtained in the previous step, 1,4-dihydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester and 1,4-dihydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester (1 h and 1 i) (7.0 g, 18.0 mmol) were added sequentially to a 1 L three-necked flask, and the mixture was heated to 80 °C and reacted for 3 hours. TLC analysis showed that the starting material had disappeared. The reaction was stopped, cooled to room temperature, and the reaction solution was concentrated under reduced pressure. The solution was then poured into a 2N sodium carbonate aqueous solution, adjusted to pH 9-10, separated, washed with water, dried over anhydrous sodium sulfate, concentrated, and column chromatography yielded 5.5 g of a white flocculent substance, with a yield of 67.6%. This white flocculent substance was a mixture of 1-bromo-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester and 1-bromo-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester.

[0169] Step 7: Preparation of 1-cyclopropyl-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester and 1-cyclopropyl-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester (1l and 1m)

[0170]

[0171] Toluene (200 mL), the mixed products obtained in the previous step, butyl 1-bromo-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate and butyl 1-bromo-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate (1j and 1k) (4.0 g, 8.8 mmol), water (10 mL), cesium carbonate (2.9 g, 18 mmol), and cyclopropylboronic acid (0.90 g, 10.0 mmol) were sequentially added to a 500 mL three-necked flask. Then, 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloromethane complex (Pd(dppf)Cl2·CH2Cl2) (0.46 g, 0.88 mmol) was added to the reaction solution. Nitrogen gas was purged three times, and the reaction was heated to 112 °C for 12 hours. TLC monitoring showed that the starting material had disappeared; the reaction was then stopped, and the mixture was cooled to room temperature. Extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and separated by column chromatography to give 2.0 g of product, with a yield of 54.8%. The product is a mixture of 1-cyclopropyl-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester and 1-cyclopropyl-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate butyl ester.

[0172] Step 8: Preparation of 1-cyclopropyl-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylic acid and 1-cyclopropyl-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylic acid (1n and 1o)

[0173]

[0174] The mixed products obtained in the previous step, 1-cyclopropyl-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate and 1-cyclopropyl-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate (1 L and 1 M) (1.9 g, 4.6 mmol), sodium hydroxide (1.3 g, 48.0 mmol), water (50 mL), and methanol (100 mL), were added to a 250 mL three-necked flask, and the mixture was heated to 70 °C and reacted for 11 hours. TLC analysis showed that the starting material had disappeared, at which point the reaction was stopped, and the mixture was cooled to room temperature. The solution was concentrated under reduced pressure, the pH was adjusted to 2 with concentrated hydrochloric acid, filtered, and dried to give a pale yellow solid (1.5 g, 4.2 mmol), with a yield of 91.2%. This product is a mixture of 1-cyclopropyl-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylic acid and 1-cyclopropyl-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylic acid.

[0175] Step Nine: Preparation of Methyl [(1-cyclopropyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetate and Methyl [(1-cyclopropyl-4-hydroxy-6-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetate (1p and 1q)

[0176]

[0177] At room temperature, the mixed products obtained in the previous step, 1-cyclopropyl-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylic acid and 1-cyclopropyl-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylic acid (1 L and 1 M) (1.4 g, 3.9 mmol), dichloromethane (12 mL), oxalyl chloride (0.75 g, 5.9 mmol), and 1 drop of DMF were added sequentially to a reaction flask, and the reaction was carried out for half an hour. When no more bubbles were released from the reaction solution, the solvent was removed by concentration under reduced pressure. Then, dichloromethane (10 mL) was added, followed by glycine methyl ester hydrochloride (0.78 g, 6.2 mmol) and diisopropylethylamine (2.5 g, 20.0 mmol). The mixture was stirred at room temperature for 1 hour. The reaction was stopped when the starting material disappeared under TLC monitoring. The reaction solution was washed successively with water, hydrochloric acid, saturated sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and separated by column chromatography to obtain 0.42 g of methyl [(1-cyclopropyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetate (yield 25.0%) and 0.61 g of methyl [(1-cyclopropyl-4-hydroxy-6-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetate (yield 36.3%).

[0178] Step 10: Preparation of [(1-cyclopropyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (1r)

[0179]

[0180] The obtained [(1-cyclopropyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-methyl acetate (1p) (0.22 g, 0.52 mmol), water (2 mL), methanol (30 mL), and anhydrous lithium chloride (100 mg) were added to a 100 mL three-necked flask and stirred overnight at room temperature. TLC analysis showed the starting material had disappeared. The mixture was concentrated under reduced pressure to remove methanol, and the pH was adjusted to 2 with 2N hydrochloric acid. The solution was filtered, washed with water, and dried to give 0.16 g of a white solid, with a yield of 75.2%. NMR data: 1 H NMR (500Hz, CDCl3) δ13.17(s,1H), δ9.03(d,1H), δ9.03(d,1H), δ8.64(d,1H), δ7.63(m,1H), δ7.5 2(m,2H),δ7.21(m,2H),δ7.19(m,1H),δ4.02(s,2H),δ2.78(m,1H),δ1.21(m,2H),δ1.02(m,1H); MS m / z:411.2(M-1).

[0181] Example 2: Preparation of [(1-cyclopropyl-4-hydroxy-6-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (1s)

[0182]

[0183] 0.20 g (1 q, 0.47 mmol) of methyl [(1-cyclopropyl-4-hydroxy-6-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetate, prepared by the aforementioned method, 2 mL of water, 30 mL of methanol, and 120 mg of anhydrous lithium chloride were added to a 100 mL single-necked flask and stirred overnight at room temperature (20-25 °C). The solid disappeared. TLC analysis showed the disappearance of the starting material. The mixture was concentrated under reduced pressure at 40 °C to remove methanol. 30 mL of water was added, resulting in a white flocculent precipitate. The precipitate was adjusted to pH 2 with 2N hydrochloric acid, ultrasonically dispersed, filtered, washed with water, and dried to obtain 0.16 g (0.39 mmol) of 1-cyclopropyl-7-(3-chlorophenoxy)-4-hydroxyisoquinoline-3-carbonylaminoacetic acid (1 s), a white solid, with a yield of 82.7%. NMR data: 1H NMR (500Hz, CDCl3) δ13.35 (s, 1H), δ9.00 (s, 1H), δ8.32 (d, 1H), δ8.07 (d, 1H), δ7.63 (d, 1H), δ7.58 (d, 1H), δ7.49 MS m / z:411.2(M-1).

[0184] Example 3: Preparation of [(1-cyclopropyl-4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (2i)

[0185]

[0186] In Example 1, 3-chlorophenol in step one was replaced with phenol, and the remaining operations were the same as in Example 1. A final yield of 0.16 g of white solid was obtained. NMR data: 1 H NMR (500Hz, CDCl3) δ13.13(s,1H), δ9.01(s,1H), δ8.59(d,1H), δ7.61(m,1H), δ7.54(m,2H), δ7.52(m,2H), δ7.5 MS m / z:377.3(M+1).

[0187] Example 4: Preparation of [(1-cyclopropyl-4-hydroxy-6-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (2j)

[0188]

[0189] In Example 2, methyl [(1-cyclopropyl-4-hydroxy-6-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetate was replaced with the intermediate [(1-cyclopropyl-4-hydroxy-6-phenoxy-isoquinoline-3-carbonyl)-amino]-acetate]-acetate obtained in Example 3, with the rest following the same procedure as in Example 2. A final product of 0.16 g was obtained as a white solid. NMR data: 1H NMR (500Hz, CDCl3) δ13.33(s,1H), δ8.98(s,1H), δ8.32(d,1H), δ7.97(m,1H), δ7.56(m,1H), δ7.5 4(m,2H),δ7.27(m,1H),δ7.21(m,1H),δ4.02(s,2H),δ2.50(m,1H),δ1.16(m,2H),δ0.96(m,2H); MS m / z:377.3(M+1).

[0190] Example 5: Preparation of [(1-methyl-4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (3g)

[0191]

[0192] Step 1: The mixture of 1-bromo-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate and 1-bromo-4-hydroxy-6-m-chlorophenoxyisoquinoline-3-carboxylate (1.5 g, 3.3 mmol), 1,4-dioxane (75 mL), potassium carbonate (2.8 g, 20.2 mmol), and trimethylboroxane (2.0 mL, 7.0 mmol) obtained in Step 6 of Example 1 was added to a 250 mL three-necked flask. Then, tetrakis(triphenylphosphine)palladium (0.30 g, 0.27 mmol) was added to the reaction solution. Nitrogen gas was purged three times, and the temperature was raised to 95 °C for 4.5 hours. The reaction was stopped when the starting material disappeared as detected by TLC. The mixture was cooled to room temperature, extracted with ethyl acetate, and the phases were separated. The organic phase was dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to give 402 mg of compound [(1-methyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (yield 31.3%) and 220 mg of compound [(1-methyl-4-hydroxy-6-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (yield 17.2%).

[0193] Step 2: The [(1-methyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (300 mg, 0.78 mmol), sodium hydroxide (140 mg, 3.5 mmol), water (25 mL), and methanol (10 mL) obtained in the previous step were added to a 100 mL single-necked flask, and the mixture was heated to 70 °C and reacted for 4 hours. TLC monitoring showed that the starting material had disappeared, at which point the reaction was stopped, and the mixture was cooled to room temperature. The solution was concentrated under reduced pressure, the pH was adjusted to 2 with concentrated hydrochloric acid, filtered, and dried to obtain 220 mg of a pale yellow solid, 1-aldyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carboxylic acid butyl ester, with a yield of 85.5%.

[0194] Step 3: At room temperature, 1-methyl-4-hydroxy-7-m-chlorophenoxyisoquinoline-3-carboxylate (220 mg, 0.67 mmol), dichloromethane (15 mL), 1 drop of DMF, and oxaloyl chloride (0.30 g, 2.36 mmol) were added sequentially to the reaction flask. The reaction was carried out for half an hour until no more bubbles were released. The solvent was removed by concentration under reduced pressure. Then, dichloromethane (15 mL), glycine methyl ester hydrochloride (0.86 g, 6.85 mmol), and diisopropylethylamine (1.4 g, 10.8 mmol) were added sequentially, and the mixture was stirred at room temperature for half an hour. TLC monitoring showed that the starting material had disappeared. The reaction was then stopped, and the mixture was cooled to room temperature. The reaction solution was washed with water, dilute hydrochloric acid, saturated sodium carbonate aqueous solution, and 5% sodium chloride aqueous solution. The phases were separated, the organic phase was dried with anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 180 mg of compound [(1-methyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-methyl acetate, with a yield of 67.0%.

[0195] Step 4: [(1-methyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid methyl ester (180 mg, 0.45 mmol), water (10 mL), methanol (10 mL), and anhydrous lithium chloride (140 mg, 5.9 mmol) were added to a 50 mL single-necked flask and stirred at room temperature for half an hour. The starting material disappeared during TLC. The mixture was concentrated under reduced pressure to remove methanol, and the pH was adjusted to 2 with 2N hydrochloric acid. The solution was filtered and dried to obtain 150 mg of a white solid [(1-methyl-4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid, with a yield of 86.3%. NMR data: 1 H NMR (500Hz, CDCl3) δ13.19 (s, 1H), δ9.16 (d, J = 8.0Hz, 1H), δ8.29 (d, J = 11.5Hz, 1H), δ7.60 (dd, J =11.5Hz,1H), δ7.52(m,2H), δ7.35(m,2H), δ7.19(m,1H), δ4.04(s,2H), δ2.84(s,3H); Lc-Ms: m / z 487.2(M+1).

[0196] Example 6: Preparation of [(1-methyl-4-hydroxy-6-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (3h)

[0197]

[0198] The [(1-methyl-4-hydroxy-7-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid in step two of Example 5 was replaced with [(1-methyl-4-hydroxy-6-m-chlorophenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid, with the rest following the same procedure as in Example 5. 114 mg of a white solid [(1-methyl-4-hydroxy-6-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid was finally obtained, with a yield of 67.0%. NMR data: 1 H NMR (500Hz, CDCl3) δ13.34 (s, 1H), δ9.13 (d, J = 8.0Hz, 1H), δ8.33 (d, J = 11.0Hz, 1H), δ7.75 (d, J = 8.0Hz, 1H), δ7.6 0(dd,J=8.5Hz,1H), δ7.49(d,J=10.0Hz,2H), δ7.30(m,1H), δ7.13(m,1H), δ4.04(s,2H), δ2.75(s,3H); Lc-Ms: m / z 487.2(M+1).

[0199] Example 7: Biological test: PHD2 enzyme activity inhibition detection

[0200] Reagents and consumables: PHD2 enzyme: purchased from Activemotif; α-ketoglutarate sodium salt: purchased from Sigma; FITC-HIF1α: purchased from GL; Nunc multiwell plates TM 384: Purchased from ThermoScientific.

[0201] Test method:

[0202] (1) Prepare 1×Assaybuffer test solution (10mM HEPES, 150mM NaCl, 0.05% Tween 20, pH 7.4); (2) Preparation of compound concentration gradient: The test concentration of the test compound starts at 10μM, is diluted 3 times, 10 concentrations are obtained, and the test is repeated in wells; the solution is diluted to a final concentration of 100 times in a 384-well plate, and then 100nL is transferred to the 384 reaction plate for later use using Echo 550; 100nL of 100% DMSO is added to the negative control well and the positive control well respectively; (3) Prepare enzyme solution with a final concentration of 2 times using 1×Assaybuffer; (4) Add 5μL of enzyme solution with a final concentration of 2 times to the compound well and the positive control well respectively; (5) Add to the negative control well (6) Centrifuge at 1000 rpm for 30 seconds, vortex and incubate for 15 min; (7) Prepare a tracer solution with a final concentration of 2 times using 1×Assaybuffer; add 5.0 μL of tracer solution with a final concentration of 2 times to start the reaction; (8) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, vortex and mix for 60 min, read the mP value from Envision (Multimode Plate Reader, PerkinElmer), export the data and process it to obtain the inhibition rate of the test compound. The results are shown in the table below:

[0203] Table 1.

[0204] Compound numbering <![CDATA[IC 50 (nM)]]> 1r 1080 1s 3719 2i 839 2j 781 3g 82.6 3h 61.2

[0205] As can be seen from the table above, the compounds in the embodiments of the present invention have excellent HIF-prolyl hydroxylase inhibitory activity.

[0206] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An isoquinoline carboxylic acid compound represented by the following formula: ###0001### or a pharmaceutically acceptable salt thereof.

2. An isoquinoline carboxylic acid compound represented by the following formula: ###0002### or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition comprising the compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

4. Use of the compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, for the manufacture of a medicament for the treatment or prophylaxis of a disease mediated by hypoxia inducible factor and / or erythropoietin, said disease being renal anemia.

5. Use of the compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 3, for the manufacture of a medicament for inhibiting the activity of prolyl hydroxylase.

6. A method for producing an isoquinolinecarboxylic acid compound or a pharmaceutically acceptable salt thereof, characterized by, The preparation method of the compound comprises the following steps: Step 1: preparing a diaryl ether represented by formula 4c from a 4-nitrophthalonitrile derivative represented by formula 4a through a nucleophilic substitution reaction; Step 2: preparing a dicarboxylic acid represented by formula 4d from the diaryl ether represented by formula 4c through cyanide hydrolysis; Step 3: preparing a ring-closed carboxylic acid represented by formula 4f from the dicarboxylic acid represented by formula 4d through ring closure with ammonia; Step 4: preparing an ester represented by formula 4g from the ring-closed carboxylic acid represented by formula 4f through ester hydrolysis; Step 5: preparing ring-extended products represented by formula 4h and 4i from the ester represented by formula 4g through a Dickman condensation reaction; Step 6: preparing bromoisoquinoline derivative esters represented by formula 4j and 4k from the ring-extended products represented by formula 4h and 4i through a bromination reaction; Step 7a: preparing three-membered ring-containing isoquinoline derivatives represented by formula 4l and 4m from the bromoisoquinoline derivative esters represented by formula 4j and 4k through a Suzuki coupling reaction; Step 8a: preparing carboxylic acids represented by formula 4n and 4o from the three-membered ring-containing isoquinoline derivatives represented by formula 4l and 4m through ester hydrolysis; Step 9a: preparing amides represented by formula 4p and 4q from the carboxylic acids represented by formula 4n and 4o through condensation with glycine methyl ester or its hydrochloride salt; Step 10a, 10b: preparing carboxylic acids represented by formula 4r and 4s from the amides represented by formula 4p and 4q, respectively, through ester hydrolysis; wherein R1', R2' are each selected from hydrogen; Ar is phenyl.

7. The production method according to claim 6, characterized by, In step (7a), the coupling reaction is achieved by a coupling agent in a mixture of toluene and water.

8. The preparation method according to claim 6, characterized in that, In step (7a), the temperature of the coupling reaction is 50-120°C.

9. The preparation method according to claim 6, characterized in that, In step (7a), the coupling agent used is a metal Pd compound.

10. The method of claim 6, wherein, In step (7a), the coupling agent used is 1,1'-bis(diphenylphosphino)ferrocene palladium(II) chloride complex Pd(dppf)Cl2·CH2Cl2.

11. The method of claim 6, wherein, In step (7a), the molar ratio of the coupling agent to the bromoisoquinoline derivative ester is 0.01-1.

12. The method of claim 6, wherein, In step (7a), the molar ratio of the coupling agent to the bromoisoquinoline derivative ester is 0.01-0.

2.

13. The preparation method according to claim 6, characterized in that, In step (7a), the molar ratio of the coupling agent to the bromoisoquinoline derivative ester is 0.05-0.

1.

14. A process for preparing an isoquinoline carboxylic acid compound or a pharmaceutically acceptable salt thereof, characterized by, The compound preparation method comprises the following steps: Step 1: preparing a diaryl ether shown in formula 4c from a 4-nitrophthalonitrile derivative shown in formula 4a by a nucleophilic substitution reaction; Step 2: preparing a dicarboxylic acid shown in formula 4d from the diaryl ether shown in formula 4c by cyanide hydrolysis; Step 3: preparing a cyclic carboxylic acid shown in formula 4f from the dicarboxylic acid shown in formula 4d by ring closure with ammonia; Step 4: preparing an ester shown in formula 4g from the cyclic carboxylic acid shown in formula 4f by ester hydrolysis; Step 5: preparing ring expansion products shown in formula 4h and 4i from the ester shown in formula 4g by a Dickman condensation reaction; Step 6: preparing brominated isoquinoline derivative esters shown in formula 4j and 4k from the ring expansion products shown in formula 4h and 4i by a bromination reaction; Step 7b: preparing tri-membered ring isoquinoline derivative esters shown in formula 5l and 5m from the brominated isoquinoline derivative esters shown in formula 4j and 4k by a Suzuki coupling reaction; Step 8b, 8c: preparing carboxylic acids shown in formula 5n and 5o from the tri-membered ring isoquinoline derivative esters shown in formula 5l and 5m by ester hydrolysis, respectively; Step 9b, 9c: preparing amides shown in formula 5p and 5q from the carboxylic acids shown in formula 5n and 5o by condensation with glycine methyl ester or its hydrochloride, respectively; Step 10c, 10d: preparing carboxylic acids shown in formula 5r and 5s from the amides shown in formula 5p and 5q by ester hydrolysis, respectively; wherein R1', R2' are both selected from hydrogen, and Ar is meta-chlorophenyl.

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