Process for the preparation of membrane iron transport protein inhibitors

Abstract

The present invention relates to a novel process for the preparation of compounds of formula (I) and pharmaceutically acceptable salts thereof, which are suitable for use as drugs for the prevention and / or treatment of diseases caused by hepcidin deficiency or iron metabolism disorders leading to increased iron levels or increased iron absorption, including iron overload, thalassemia, sickle cell disease and hemochromatosis.

Description

[0001] Foreword

[0002] This invention relates to a novel method for preparing compounds of formula (I) and their pharmaceutically acceptable salts.

[0003]

[0004] The compounds of general formula (I) of the present invention, as inhibitors of membrane iron transporters, are therefore particularly suitable as medicines for the prevention and / or treatment of diseases caused by hepcidin deficiency or iron metabolism disorders leading to increased iron levels or increased iron absorption. The compounds of general formula (I) of the present invention are also particularly suitable for the prevention and / or treatment of iron overload, including thalassemia, sickle cell disease, and hemochromatosis, as well as for the prevention and / or treatment of diseases associated with or caused by increased iron levels, increased iron absorption, or iron overload.

[0005] Background and Existing Technology

[0006] Iron is an essential micronutrient for almost all organisms, and is particularly relevant to growth and blood formation. The balance of iron metabolism in this context is primarily regulated by the iron reuptake from hemoglobin in aging red blood cells and the level of dietary iron absorption in the duodenum. Released iron is absorbed in the intestines, particularly via specific transport systems (DMT-1, ferroportin), and then transported to appropriate tissues and organs (transferrin, transferrin receptors).

[0007] Mammalian organisms cannot actively excrete iron. Iron metabolism is primarily controlled by hepcidin (a peptide hormone produced in the liver), which is released cellularly from iron in macrophages, hepatocytes, and intestinal cells. Hepcidin acts on iron absorption through the intestine and placenta, and on the release of iron from the reticuloendothelial system. In vivo, hepcidin is synthesized in the liver by a gene called hepcidinogen, encoded by a gene called the HAMP gene. Hepcidin formation is directly regulated in relation to the organism's iron levels; more hepcidin is formed if the organism is supplied with sufficient iron and oxygen, and less is formed if iron and oxygen levels are low, or in cases of increased erythropoiesis. In the small intestinal mucosal cells and macrophages, hepcidin binds to membrane iron transporters, which normally transport circulating iron from inside the cell into the bloodstream.

[0008] Membrane siderotransporters are transmembrane proteins composed of 571 amino acids, formed in the liver, spleen, kidneys, heart, intestine, and placenta. In particular, membrane siderotransporters are located in the basolateral membrane of intestinal epithelial cells. Therefore, membrane siderotransporters bound in this manner function to transport iron into the bloodstream. In this case, membrane siderotransporters are most likely to transport iron as Fe... 2+ Iron is transported in the form of hepcidin. If hepcidin binds to a membrane iron transporter, the membrane iron transporter is transported into the cell, where it is broken down, almost completely blocking the release of iron from the cell during the phagocytic cycle. If the membrane iron transporter is inactivated, for example by hepcidin, preventing it from exporting iron stored in mucosal cells, the stored iron is lost through natural excretion in feces. Therefore, when membrane iron transporters are inactivated or inhibited, for example by hepcidin, iron absorption in the intestine is reduced. Furthermore, membrane iron transporters are prominently located in the reticuloendothelial system (RES), to which macrophages also belong. On the other hand, if serum iron levels are low, hepcidin production in hepatocytes decreases, resulting in less hepcidin release and therefore less inactivated membrane iron, allowing a larger amount of stored iron to be transported into the serum.

[0009] Therefore, the hepcidin-membrane iron transporter system directly regulates iron metabolism, and thus, disruptions to the hepcidin regulatory mechanism have a direct impact on iron metabolism in organisms. In principle, the hepcidin-membrane iron transporter regulatory mechanism operates through the following two opposing principles:

[0010] On the one hand, increased hepcidin leads to the inactivation of membrane iron transporters, thereby blocking the release of stored iron from cells into the serum and thus reducing serum iron levels. Under pathological conditions, decreased serum iron levels lead to decreased hemoglobin levels and reduced red blood cell production, resulting in iron deficiency anemia.

[0011] On the other hand, a decrease in hepcidin leads to an increase in active membrane iron transporters, thereby increasing the release of stored iron and, for example, iron intake from food, thus increasing serum iron levels. In pathological conditions, increased iron levels lead to iron overload.

[0012] Iron overload and related diseases are characterized by excessive iron levels. The problem stems from excessive serum iron levels, leading to non-transferrin-bound iron (NTBI). NTBI is rapidly and non-specifically absorbed by organs, resulting in iron accumulation in tissues and organs. Iron overload can cause numerous diseases and undesirable medical conditions, including damage to the heart, liver, and endocrine system. Furthermore, iron accumulation in the brain has been observed in patients with neurodegenerative diseases such as Alzheimer's and Parkinson's. As a particularly harmful aspect of excessive free iron, the formation of undesirable free radicals must be mentioned. Iron(II) ions, in particular, catalyze the formation of reactive oxygen species (ROS) (especially via the Fenton reaction). These ROS cause damage to DNA, lipids, proteins, and carbohydrates, with profound effects on cells, tissues, and organs, and are well-known, with their so-called oxidative stress described in the literature.

[0013] In addition to using chelating agents such as deferoxamine (also known as deferoxamine B, N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutyryl}amino)pentyl]-N-hydroxysuccinamide or deferasirox () 4-(3,5-bis(2-hydroxyphenyl)-1H-1,2,4-triazol-1-yl)benzoic acid) and deferoxone ( In addition to conventional methods of treating iron overload by removing iron from the body (3-hydroxy-1,2-dimethylpyridin-4(1H)-one), compounds that act as hepcidin agonists or have inhibitory or supportive effects on biochemical regulatory pathways in iron metabolism, such as hepcidin mimic peptides, have been described. These treatments are based on directly engaging in the disrupted iron metabolic pathways by providing hepcidin mimics or agonists, i.e., acting as a hepcidin substitute or supplier, through the direct action of the primary regulator, hepcidin. This method is based on the therapeutic principle of treating iron overload (i.e., excessive serum iron levels) by inhibiting membrane iron transporters, thereby blocking excessive iron absorption through the hepcidin inactivation mechanism.

[0014] Membrane iron transporter inhibitors according to formula (I) and methods for their preparation have been described in WO2017 / 068089 and WO2017 / 068090. The preparation methods described therein provide 12 steps comprising several chromatographic steps, resulting in an inefficient preparation method that is time-consuming, costly, and labor-intensive. A further feature of the described methods is the relatively low yield, and some of these steps raise safety and technical issues due to the formation of critical byproducts.

[0015] In addition, international application WO2018 / 192973 describes the preparation and crystallization of various specific salts of selected membrane iron transporter inhibitors as described therein and as described in WO2017 / 068089 and WO2017 / 068090.

[0016] WO2011 / 029832 relates to thiazole and oxazole compounds used as hepcidin antagonists, which are described as suitable for the treatment of iron deficiency diseases, and describes a method for preparing said compounds according to synthetic route 3). The described method also includes multiple steps with chromatographic separation and purification steps, and is therefore disadvantageous from an efficiency standpoint.

[0017] ACVeronese et al., “One-Pot Synthesis of 2-Vinylimidazole Derivatives by Reaction of α-Hydroxyimino-β-dicarbonyl Compounds with Allylamine” (1985), described a one-pot synthetic reaction to obtain 2-vinylimidazole derivatives, but did not mention the compounds according to formula (I), (II) or (II') or the selected intermediates of the present invention or their preparation methods.

[0018] Purpose

[0019] The object of this invention is to provide a novel method for preparing selected membrane iron transporter inhibitors and their pharmaceutically acceptable salts as defined by general formula (I) of this invention. This novel method should be improved in at least one aspect by increasing yield, process efficiency, reducing process steps, improving resource utilization, for example by using commercially available or cheaper starting compounds or by using starting compounds and intermediate compounds that can be prepared in a time-saving, cost-saving, and labor-saving manner; by minimizing chromatographic steps; by improving operational safety; by avoiding critical or harmful byproducts; by avoiding critical reaction components such as Sn reagents; and by minimizing intermediate separation steps. Another object of this invention is to provide a novel method that provides membrane iron transporter inhibitor compounds with improved impurity distribution and / or higher purity compared to compounds obtainable by known methods.

[0020] Therefore, another object of the present invention relates to providing membrane iron transporter inhibitor compounds with high purity and improved impurity distribution.

[0021] On the other hand, it involves providing novel membrane iron transporter inhibitor compounds, which has been solved with new compounds according to formula (II').

[0022] This objective has been achieved by providing new and improved methods for preparing selected membrane iron transporter inhibitors as defined by general formula (I) of the present invention and their pharmaceutically acceptable salts. Detailed Implementation

[0023] The first aspect of the present invention provides a novel method for preparing compounds of general formula (I).

[0024]

[0025] Includes the following steps:

[0026] The compound of formula (IM-3) is reacted with the compound of formula (RM-3).

[0027]

[0028] To provide compounds of formula (I);

[0029] in

[0030] X 1 It is N, S, or O; and

[0031] X 2 It is N, S, or O;

[0032] The condition is that X 1 and X 2 One of them is N, and X 1 and X 2 different;

[0033] m is an integer of 1, 2, or 3;

[0034] n is an integer of 1, 2, 3 or 4;

[0035] o is an integer of 1, 2, 3 or 4;

[0036] A represents a CH group, a CH2-CH group, or a CH2-CH2-CH group;

[0037] R 1 and R 2 Selected independently

[0038] -hydrogen, and

[0039] -C1-C4 alkyl groups, which can be substituted by one or two substituents;

[0040] R 3 This indicates 0, 1, 2, or 3 substituents, which can be independently selected.

[0041] -halogen,

[0042] -Cyano

[0043] -C1-C4 alkyl,

[0044] -C1-C3 haloalkyl;

[0045] -C1-C4 alkoxy groups, and

[0046] -Carboxyl group;

[0047] R 4 Selected from

[0048] -hydrogen,

[0049] -halogen,

[0050] -C1-C3 alkyl groups, and

[0051] -C1-C3 haloalkyl;

[0052] R 5 Selected from

[0053] -Aryl groups can have 1 to 3 substituents, and

[0054] - Can be a monocyclic or bicyclic heteroaryl group with one to three substituents; and

[0055] R 6 Selected from

[0056] -hydrogen,

[0057] -halogen,

[0058] -C1-C4 alkyl groups, which can be substituted by one or two substituents;

[0059] -C1-C3 haloalkyl.

[0060] definition

[0061] The term "substituted" means that one or more hydrogen atoms on a specified atom or group are selectively replaced by a group selected from the specified group, provided that the replacement does not exceed the normal valence of the specified atom under its existing condition. Combinations of substituents and / or variables are permitted.

[0062] The terms "optionally substituted" or "optionally substituented" mean that the number of substituents may be equal to or not equal to zero. Unless otherwise stated, an optionally substituted group may be substituted with as many optional substituents as possible, which can be tuned by replacing hydrogen atoms with non-hydrogen substituents on any available carbon or nitrogen atom. Typically, the number of optional substituents, when present, can be 1, 2, 3, 4, or 5, especially 1, 2, or 3.

[0063] As used herein, the terms “one or more” or “one or more”, for example in the definition of substituents in compounds of general formula (I) of the present invention, mean “1, 2, 3, 4 or 5, particularly 1, 2, 3 or 4, more particularly 1, 2 or 3, even more particularly 1 or 2”.

[0064] When used in the claims or specification, the term "comprising" includes "consisting of".

[0065] If any item in this specification is referred to as “as mentioned herein” or “as defined herein (anywhere)”, it means that it may be mentioned anywhere in this specification or may have the meaning as defined anywhere in this specification.

[0066] The terms used in this specification have the following meanings:

[0067] The term "halogen" or "halogen atom" refers to a fluorine, chlorine, bromine, or iodine atom, especially a fluorine, chlorine, or bromine atom, with a preferred choice involving chlorine or fluorine, a further preferred choice involving bromine or fluorine, and fluorine being the most preferred.

[0068] The term "C1-C4 alkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon group having one, two, three, or four carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl groups, or their isomers. The term "C1-C3 alkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon group having one, two, or three carbon atoms, such as methyl, ethyl, n-propyl, or isopropyl groups.

[0069] C1-C4 or C1-C3 alkyl groups may optionally be substituted with one or two substituents, preferably with one substituent. Such optional substituents are preferably selected from: halogens (forming halogen-substituted C1-C4 or C1-C3 alkyl groups as defined below), preferably C3-C6 cycloalkyl groups containing 3, 4, 5, or 6 carbon atoms (e.g., preferably cyclopropyl as defined below), monocyclic or bicyclic heteroaryl groups (e.g., preferably benzimidazolyl groups), amino groups as defined below, carboxyl groups, and aminocarbonyl groups as defined below.

[0070] The term "C1-C3 haloalkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon group, wherein the term "C1-C3 alkyl" has the meaning as defined above, and wherein one or more hydrogen atoms are replaced by halogen atoms, either identically or differently. Specifically, the halogen atom is a fluorine atom. More specifically, all of the halogen atoms are fluorine atoms ("C1-C3 fluoroalkyl"). The C1-C3 haloalkyl group is, for example, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, 3,3,3-trifluoropropyl, or 1,3-difluoropropyl-2-yl, wherein a trifluoromethyl group is particularly preferred.

[0071] The term “C1-C4 alkoxy” refers to a straight-chain or branched saturated monovalent group of the formula (C1-C4 alkyl)O, wherein the term “C1-C4 alkyl” is as defined above, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, or tert-butoxy groups, or isomers thereof, wherein the methoxy group is particularly preferred.

[0072] The term "carboxyl group" refers to the group [-(C=O)-OH].

[0073] The term "aminocarbonyl group" refers to the group [NH2-(C=O)-].

[0074] The term "amino group" includes amino (-NH2), mono- or dialkylamino (alkyl-NH-, (alkyl)2N-), wherein the definition of "alkyl" is given above for C1-C4-alkyl and C1-C3-alkyl. Preferred groups are amino (-NH2) and mono- or dimethylamino. Most preferred is the amino (-NH2) group.

[0075] The term "aryl" includes aromatic residues containing 6 to 14 carbon atoms (excluding carbon atoms of possible substituents), which may be monocyclic or bicyclic, including, for example, phenyl, naphthyl, phenanthryl, and anthracene, which may optionally be substituted by one, two, or three identical or different substituents selected from hydroxyl, halogens as defined above (e.g., preferably F, Br, and Cl), cyano, carboxyl groups as defined above, amino groups as defined above, C1-C4-alkyl or C1-C3-alkyl groups as defined above (e.g., preferably methyl), C1-C3 haloalkyl groups as defined above (e.g., preferably trifluoromethyl), and C1-C4-alkoxy groups as defined above (e.g., preferably methoxy).

[0076] Preferred phenyl groups are optionally substituted, such as unsubstituted phenyl groups and phenyl groups substituted with one to three, more preferably one or two, substituents, which may be the same or different. The one to three phenyl substituents are specifically selected from groups as defined above.

[0077] The term "monocyclic or bicyclic heteroaryl" includes heteroaromatic residues containing 4 to 9 ring carbon atoms, which preferably also contain 1 to 3 identical or different heteroatoms selected from the series S, O, N in the ring, thus preferably forming 5- to 12-membered heteroaromatic residues, which are preferably monocyclic but can also be bicyclic. Preferred aromatic heterocyclic residues include: pyridinyl (nitrophenyl), pyridinyl-N-oxide, pyridazinyl, pyrimidinyl, pyrazinyl, thiopheneyl (thiopheneyl), furanyl, pyrroleyl, pyrazolyl, imidazolyl, triazolyl, thiazolyl, oxazolyl or isoxazolyl, inazinyl, indolyl, benzo[b]thiopheneyl, benzo[b]furanyl, inzolyl, quinolinyl, isoquinolinyl, naphridinyl, quinazolinyl, quinoxalinyl. Preferably, a 5- or 6-membered aromatic heterocycle is used, for example, a group selected from 5-membered heteroaryl groups, such as thiazolyl (e.g., thiazolyl-2-yl, 2-thiazolyl-2-yl, 2-thiazolyl-4-yl), thienyl (e.g., thienyl-3-yl), pyrazolyl (e.g., 1-pyrazolyl-4-yl, 3-pyrazolyl-5-yl), imidazole (e.g., imidazole-2-yl, 2-imidazole-4-yl, 1-imidazole-4-yl), triazolyl (e.g., 1-triazolyl-3-yl, 1-triazolyl-4-yl, e.g., 1,2,4-triazolyl-3-yl or 1,2,3-triazolyl-4-yl), oxazolyl (e.g., 2-oxazolyl-4-yl, 2-oxazolyl-5-yl), oxadiazolyl (e.g., 1,2... ,4-oxadiazol-3-yl); and groups selected from 6-membered heteroaryl groups, such as pyridyl (aziridine) (e.g., pyridin-1-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 2-pyridin-4-yl, 2-pyridin-6-yl, 3-pyridin-5-yl (aziridine-1-yl, aziridine-2-yl, aziridine-3-yl, aziridine-4-yl, 2-aziridine-4-yl, 2-aziridine-6-yl, 3-aziridine-5-yl)), pyrimidine-2-yl, pyrimidine-4-yl, pyrimidine-5-yl; and groups selected from bicyclic heteroaryl residues, especially benzimidazolyl, such as benzimidazol-2-yl, benzimidazol-4-yl, benzimidazol-5-yl.

[0078] The aforementioned heteroaryl groups may have one or more (preferably one, two or three, more preferably one or two) identical or different substituents, which are particularly selected from hydroxyl, halogens as defined above (e.g., preferably F, Br and Cl), cyano, carboxyl groups as defined above, amino groups as defined above, C1-C4-alkyl or C1-C3-alkyl groups as defined above (e.g., preferably methyl), C1-C3-haloalkyl groups as defined above (e.g., preferably trifluoromethyl), and C1-C4-alkoxy groups as defined above (e.g., preferably methoxy).

[0079] Particularly preferred monocyclic or bicyclic heteroaryl groups are pyridyl groups, which have one, two, or three substituents selected from the groups defined above. Preferably, the pyridyl group has one or two substituents, more preferably one substituent. The one, two, or three optional pyridyl-substituents are preferably selected from: C1-C3-alkyl groups as defined above (e.g., particularly methyl), halogens as defined above (e.g., particularly fluorine and bromine (of which fluorine is most preferred)), and C1-C3-haloalkyl groups as defined above (e.g., particularly trifluoromethyl). Most preferably, a substituent forming a group represented by the following formula (denoted as R) is preferred. y )

[0080]

[0081] Where * represents the bonding location and R y The substituent is selected from the following: C1-C3-alkyl (e.g., preferably methyl), halogen (e.g., preferably fluorine or bromine) and C1-C3-haloalkyl (e.g., preferably trifluoromethyl), wherein fluorine or bromine is more preferred, and fluorine is most preferred. Summary of the Invention

[0082] In a first aspect, the present invention provides a novel method for preparing compounds of general formula (I) as defined herein, wherein an intermediate compound of formula (IM-3) is reacted with a compound of formula (RM-3) to provide a compound of formula (I):

[0083]

[0084] Where "A" represents a CH group, a CH2-CH group, or a CH2-CH2-CH group, depending on the [ ] m The defined desired alkylene chain length. Using a CH group as group "A" will result in a chain length of m=1. Using a CH2-CH group as group "A" will result in a chain length of m=2. Using a CH2-CH2-CH group as group "A" will result in a chain length of m=3.

[0085] The method steps are preferably carried out at a high temperature between 30°C and 90°C under alkaline conditions. Alkaline conditions can be achieved by adding a suitable alkali, including, for example, inorganic and organic alkalis mentioned below. Preferred alkalis are lithium hydroxide and sodium hydroxide.

[0086] In a second aspect of the invention, the novel method may further include the additional step of preparing an intermediate compound of formula (IM-3) by reacting an intermediate compound of formula (IM-2) with a compound of formula (RM-2).

[0087]

[0088] Where X 1 X 2 o, A, R 1 R 4 and R 5 It has the meaning defined above.

[0089] The method steps for preparing the intermediate compound (IM-3) can be performed prior to the method steps for preparing compound (I) as described above. The method steps are preferably carried out using methylmorpholine and ethyl chloroformate in DCM (dichloromethane). The reaction is preferably carried out under cooling, and even more preferably at a temperature below 10°C.

[0090] The compound (RM-2) used in the method steps is preferably in the form of a salt, for example, in particular, an HCl salt.

[0091] The resulting intermediate compound (IM-3) can be extracted with an aqueous HCl solution at pH 1 and crystallized from the water, followed by separation of the intermediate compound (IM-3) by conventional filtration and drying steps.

[0092] In a third aspect of the invention, the novel method may further include the additional step of preparing an intermediate compound of formula (IM-2) by converting compound (RM-1) to compound (IM-1) and then performing ester cleavage.

[0093]

[0094] Where R y Indicates hydrogen or halogen, such as preferably chlorine, and X 1 X 2 A and R 4 It has the meaning as defined anywhere in this document.

[0095] The ester cleavage can be carried out using conventional methods via ester hydrolysis. Preferably, the ester cleavage of compound (IM-1) is carried out using a suitable base, including, for example, inorganic and organic bases mentioned below. Preferred bases are lithium hydroxide and sodium hydroxide.

[0096] The reaction can be carried out in any suitable solvent, including those listed below. THF (tetrahydrofuran) and water are preferred, and the reaction is preferably carried out at room temperature (23°C ± 3°C).

[0097] In a fourth aspect of the invention, the novel method may include the step of preparing an intermediate compound of formula (IM-1) according to the following reaction scheme a):

[0098]

[0099] Where X 1 X 2 A and R 4 It has the meaning as defined anywhere in this document.

[0100] The method steps for preparing compound (IM-1) are preferably carried out at a high temperature >40°C. Any suitable solvent can be used, including those listed below. Preferably, the reaction is carried out in THF.

[0101] Furthermore, suitable catalysts are used for the reaction, including, for example, Pd(PPh3)4, Pd2(dba)3, Pd(OAc)2 / PPh3, Pd(dppf)Cl2×DCM, and Pd / C. Pd(PPh3)4 (tetra(triphenylphosphine)palladium(0)) is particularly preferred as the catalyst in the reaction.

[0102] Alternatively, (IM-1) can be prepared by directly using a brominated olefin in gaseous form or in solution form of a commercially available organic solvent.

[0103] In another aspect, by passing R in y The compound with the Cl symbol (RM-1) is converted into compound (IM-1), and then esterified as described above to prepare the intermediate compound of formula (IM-2):

[0104]

[0105] Where X 1 X 2 A and R 4 It has the meaning as defined anywhere in this document.

[0106] Specifically, according to reaction scheme b), the method steps for preparing (IM-1) can be carried out using tributyl(vinyl)tin, as described in step 1-a' below:

[0107]

[0108] In another alternative aspect of the invention, the step of preparing the intermediate compound of formula (IM-1) is to react compound (RM-1) (wherein R) according to reaction scheme c). y (with the meaning of Cl) reacts with vinylboronic acid pinacol ester to form compound (IM-1):

[0109]

[0110] Among them, X 1 X 2 A and R 4 It has the meaning as defined anywhere in this document.

[0111] This method is advantageous because it does not require tin (Sn) reagents, which results in less environmental damage, lower costs, and lower health risks to personnel performing the method.

[0112] In a fifth aspect of the invention, compound IM-1 is prepared from compound RM-1, wherein R y The hydrogen is present, as described above. Subsequently, compound IM-1 is converted to intermediate compound IM-2 via ester cleavage, and the reaction is carried out in a single-pot, combined step. This telescoped reaction scheme has the advantage of increased efficiency because it involves fewer intermediate separation and purification steps. It significantly reduces reaction time and workload, and avoids several chromatographic steps.

[0113] In a sixth aspect of the invention, compound IM-3 is prepared from intermediate compound IM-2 formed in situ by compound IM-1 and added compound RM-2, and the reaction is carried out in a one-pot combination (condensation) reaction. This condensation reaction scheme has the advantage of improved efficiency due to fewer intermediate separation and purification steps. It significantly reduces reaction time and workload, and avoids several chromatographic steps.

[0114] In a seventh aspect of the invention, compound IM-3 is prepared via a further condensation reaction, wherein compound RM-1 is converted to intermediate compound IM-1 by a method similar to that described above, subsequently converted to intermediate compound IM-2 via ester cleavage, and then converted to intermediate compound IM-3 by adding compound RM-2. This condensation reaction scheme has the advantage of further improving efficiency due to the further reduction of intermediate separation and purification steps. It can further reduce reaction time and workload, and avoid chromatographic steps.

[0115] In an eighth aspect of the invention, compound (I) is prepared by two condensation reaction steps, wherein the first condensation reaction step corresponds to the preparation of intermediate compound IM-3 described in the seventh aspect above, and the second condensation reaction step includes converting intermediate compound IM-3 in situ into compound (I), followed by salt formation to obtain a preferred salt of compound (I) of the present invention.

[0116] In a preferred aspect of the invention, the method for preparing compound (I) is as described anywhere herein, and the free base of compound (I) is obtained by phase separation (solvent extraction) or direct separation of the resulting oil product phase (oil separation).

[0117] In a preferred aspect, the present invention relates to a method for preparing compounds of general formula (I) as described herein, wherein the substituent R 5 This indicates a monocyclic heteroaryl group, which may have one to three substituents, as defined above. Wherein, R... 5 One, two, or three optional substituents may be independently selected from: C1-C3 alkyl groups as defined above, such as preferably methyl; halogens as defined above, such as preferably fluorine or bromine (wherein fluorine is more preferred); and C1-C3 haloalkyl groups as defined above, such as preferably trifluoromethyl.

[0118] In a particularly preferred aspect, the present invention relates to a method for preparing compounds of general formula (I) as described herein, wherein the substituent R 5 It is the group represented by the following

[0119]

[0120] Where * represents the bonding location and R y Selected from: C1-C3 alkyl groups as defined above, preferably methyl; halogens as defined above, preferably fluorine or bromine (of which fluorine is more preferred); and C1-C3 haloalkyl groups as defined above, preferably trifluoromethyl. Of particular preference is R. y It is fluorine or bromine (fluorine being more preferred).

[0121] In another particular aspect, the present invention provides a novel method for preparing compounds of general formula (I) as defined herein, comprising the following reaction steps:

[0122] Step 1:

[0123]

[0124] Step 2:

[0125]

[0126] Step 3:

[0127]

[0128] Where X 1 X 2 R 3 R 4 R 6 R y A, m, n, and o have the meanings defined anywhere in this document.

[0129] Steps 1, 2, and 3 of the method are preferably performed under the conditions described above.

[0130] Preferably, reaction steps 1 and 2 are carried out in a single-pot reaction step.

[0131] A further preferred aspect of the invention relates to a novel method for preparing compounds of formula (I), wherein one or more of the following conditions are satisfied:

[0132] - The substituent "A" represents a CH group, and m represents 1; and / or

[0133] -o indicates 1; and / or

[0134] -R 1 and R 2 Each represents hydrogen; and / or

[0135] -R 4 Indicates hydrogen; and / or

[0136] -R 6 Indicates hydrogen; and / or

[0137] -R 3 Indicates hydrogen; and / or

[0138] -X 1 It is N, and X 3 It is O or S, thus forming a group.

[0139]

[0140] Or X 1 It is O or S, and X 2 It is N, thus forming a group.

[0141]

[0142] In each case, * indicates the bonding position with the carbonyl group, ** indicates the second bonding position, and R 4 Independently possessing the meaning as defined anywhere in this document;

[0143] Preferably, X1 It is N, and X 3 It is O or S, forming a functional group.

[0144]

[0145] In each case, * indicates the bonding position with the carbonyl group, and ** indicates the second bonding position, R 4 Independently possessing the meaning as defined anywhere in this document;

[0146] More preferably, X 1 It is N, and X 3 It is O, thus forming a group.

[0147]

[0148] Where * indicates the bonding position with the carbonyl group, and ** indicates the second bonding position, and R 4 It has the meaning as defined anywhere in this document.

[0149] A particularly preferred aspect of the invention relates to a novel method for preparing compounds of general formula (I) as defined herein, comprising reaction steps 1, 2, and 3 as defined above, wherein

[0150] X 1 N represents N;

[0151] X 2 Represents O;

[0152] A represents the CH group;

[0153] m represents 1;

[0154] n represents 2; and

[0155] o represents 1.

[0156] Another preferred option is

[0157] R y Represents halogen atoms; and

[0158] R 4 Selected from

[0159] -hydrogen,

[0160] - As defined above, halogens are preferably chlorine.

[0161] - C1-C3 alkyl groups as defined above, preferably methyl, and

[0162] -C1-C3 haloalkyl groups as defined above, preferably trifluoromethyl.

[0163] The preferred one is

[0164] R y Indicates fluorine or bromine (of which fluorine is preferred); and

[0165] R 3 R 4 and R 6 Each represents hydrogen.

[0166] Therefore, a preferred aspect of the invention relates to the method as described herein, wherein the compound (RM-1) is represented by formula (RM-1-a):

[0167]

[0168] And / or the compound (IM-1) therein is represented by formula (IM-1-a):

[0169]

[0170] And / or the compound (RM-2) therein is represented by the formula (RM-2-a) or (RM-2-a'):

[0171]

[0172] Preferred form of HCl salt:

[0173]

[0174] And / or the compound (RM-3) therein is represented by formula (RM-3-a):

[0175]

[0176] The method of the present invention is particularly preferred for the preparation of compounds of formula (II):

[0177]

[0178] Or compounds of formula (II'):

[0179]

[0180] And their pharmaceutically acceptable salts, especially those described herein.

[0181] Therefore, a particular aspect of the present invention relates to a method for preparing compounds of formula (II) or (II') and their pharmaceutically acceptable salts, comprising the following method steps:

[0182] Step 1-a:

[0183]

[0184] Step 2-a or 2-a' are respectively:

[0185] (Step 2-a):

[0186]

[0187] or

[0188] (Step 2-a'):

[0189]

[0190] Step 3-a or 3-a' are respectively:

[0191] (Step 3-a):

[0192]

[0193] or

[0194] (Step 3-a'):

[0195]

[0196] Preferably, reaction steps 1-a and 2-a are carried out in a single-pot reaction step.

[0197] The preferred method conditions, base, solvent, and catalyst described above are preferably used in the methods described herein.

[0198] In another aspect of the invention, the particular method includes the following alternative (but less preferred) method step 1-a', starting from compound RM-1, wherein R y It is chlorine and R 4 It is hydrogen, referred to herein as RM-1-a', followed by method steps 2-a and 3-a as described above for the preparation of compounds of formula (II) or (II') and their pharmaceutically acceptable salts:

[0199] Step 1-a ' ：

[0200]

[0201] In another aspect of the invention, it is particularly preferred that the intermediate compound (IM-1-a) is prepared by the following alternative (preferred) method step 1-a”, starting from compound RM-1, wherein R y It is chlorine, and R 4 It is hydrogen, represented in this article as RM-1-a':

[0202] Step 1-a”:

[0203]

[0204] As described above, the resulting intermediate compound (IM-1-a) can be used in subsequent method steps described herein to prepare compounds of formula (II) or (II') and their pharmaceutically acceptable salts, for example, particularly by esterifying compound (IM-1-a) to form compound (IM-2-a), followed by method steps 2-a and 3-a as described above.

[0205] Generally, a wide range of inorganic and organic bases, including lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, lithium carbonate, sodium carbonate, potassium carbonate, calcium carbonate, sodium fluoride, and potassium fluoride, can be used in the methods of the present invention as described anywhere herein. Sodium hydroxide, lithium hydroxide, potassium hydroxide, sodium carbonate, and potassium carbonate are preferred. Lithium hydroxide and sodium hydroxide are most preferred.

[0206] Furthermore, a wide variety of solvents can generally be used in the methods of the invention as described anywhere herein, including, for example, methanol, ethanol, propanol, butanol, ethyl acetate (EtOAc), propyl acetate, isopropyl acetate, acetonitrile, butyronitrile, heptane, cyclohexane, methylcyclohexane, dichloromethane (DCM), toluene, xylene, chlorobenzene, dichlorobenzene, 1,4-dioxane, tetrahydrofuran (THF), 2-methyltetrahydrofuran, 2,5-dimethyltetrahydrofuran, methyl-tert-butyl ether, cyclopentyl-methyl ether, N,N-dimethylformamide, water, and mixtures thereof. Ethanol, ethyl acetate, isopropyl acetate, dichloromethane (DCM), tetrahydrofuran (THF), water, and mixtures thereof are preferred. The solvents used in the examples below are particularly preferred.

[0207] In another aspect of the invention, a method for preparing compounds of general formula (I) or (II') as described anywhere herein includes the additional step of converting compounds of formula (I) or (II) or (II') into their pharmaceutically acceptable salts or solvates using a suitable base or acid and / or solvent.

[0208] Preferably, the compound of formula (I) or (II) or (II') is converted into a pharmaceutically acceptable salt using an acid selected from the following: benzoic acid, citric acid, fumaric acid, hydrochloric acid, lactic acid, malic acid, maleic acid, methanesulfonic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, and toluenesulfonic acid. Particularly preferred acids are selected from hydrochloric acid, sulfuric acid, and phosphoric acid, with hydrochloric acid being the most preferred.

[0209] More preferably, the compound of formula (I) or (II) or (II') is converted into a pharmaceutically acceptable salt, wherein the ratio of compound (I) or (II) or (II'): acid is 1 to 2: 1 to 3.

[0210] In principle, crystallization into a trichloride (3HCl) or a monochloride (1HCl) is possible. However, crystallization into a monochloride is less preferred in these methods because it requires further processing steps on the compound of formula (I), (II), or (II'), including, for example, solvent exchange and several solvent extraction steps. This is less advantageous considering the economics of the process. Therefore, conversion to a trichloride (3HCl) salt is most preferred.

[0211] In another aspect of the invention, the method described anywhere herein includes the step of converting a compound of formula (I) or (II) or (II') into a sulfate by adding sulfuric acid and crystalline sulfate.

[0212] Converting compounds of formula (I), (II), or (II') to hydrochloride salts provides a simple and efficient crystallization method that allows direct crystallization of the decantate phase without phase separation. Chlorinated solvents may pose adverse safety risks to humans and the environment. However, the preferred method steps for converting compounds of formula (I), (II), or (II') to hydrochloride salts have the potential for continuous processing, which is also advantageous in terms of method efficiency.

[0213] Typically, the conversion of compounds of formula (I), (II), or (II') into salts can be carried out by conventional crystallization methods. Preferably, crystallization is performed by cooling a reaction mixture comprising the reaction product and a compound of formula (I), (II), or (II') to room temperature, adding a water-miscible solvent (e.g., preferably ethanol), and adding a selected acid to form the corresponding acid salt. Preferably, crystallization is carried out at an elevated temperature, preferably below the boiling points of the organic solvent and water. The resulting crystalline salt is cooled to below room temperature and separated by conventional methods, including, for example, filtration, washing, and drying.

[0214] The formation of salts of the compounds of the present invention can be particularly carried out by the method described in international application WO2018 / 192973.

[0215] As described above, solvents used for crystallization include acetonitrile, dichloromethane (DCM), alcohols (e.g., particularly methanol, ethanol, 2-propanol (isopropanol)), aldehydes, ketones (particularly acetone), ethers (e.g., tetrahydrofuran (THF) or dioxane), esters (e.g., ethyl acetate), or alkanes (e.g., particularly pentane, hexane, heptane, or cyclohexane), or water, and mixtures thereof. Preferred solvents for crystallization are selected from acetonitrile, dichloromethane, methanol, ethanol, 2-propanol, ethyl acetate, THF, water, and mixtures thereof.

[0216] Particularly preferred solvents for crystallization are selected from acetonitrile, methanol, ethanol, 2-propanol, ethyl acetate, THF, water, and mixtures thereof. Preferred water / solvent mixtures include mixtures of water and acetone, mixtures of water and ethanol, and mixtures of water and methanol, wherein mixtures of water and ethanol and mixtures of water and methanol are preferred.

[0217] Particularly preferred solvents for crystallization are those selected from acetonitrile, dichloromethane, ethanol, 2-propanol (isopropanol), acetone, and ethyl acetate, as well as mixtures thereof with water, particularly mixtures of ethanol and water and mixtures of acetone and water. Solvents described in the following examples are particularly preferred.

[0218] Particularly preferred mixtures are those of the following solvents and water (the ratio of solvent mixtures given anywhere in this document is always volume:volume):

[0219] - Acetone:water = 9:1 (volume:volume)

[0220] - Acetone:water = 95:1 (volume:volume)

[0221] - Ethanol:Water = 4:1 (volume:volume)

[0222] - Ethanol:Water = 3:1 (volume:volume)

[0223] - Ethanol:Water = 8:2 (volume:volume).

[0224] It is particularly preferred that compound (I) or (II) or (II') be converted into a salt in a one-pot reaction of condensation, while the intermediate compound IM-3 reacts with compound RM-3 to form compound (I) or (II) or (II').

[0225] Salts of compounds of formula (I), (II), or (II') may exist in amorphous, polymorphic, crystalline, and / or semi-crystalline (partially crystalline) forms, as well as in the form of solvates (or hydrates) of the salt. Preferably, the salts of the present invention exist in crystalline and / or semi-crystalline (partially crystalline) forms and / or in the form of solvates (hydrates).

[0226] The preferred crystallinity of the salts or salt solvates of the present invention can be determined using conventional analytical methods, particularly by using various X-ray methods, which makes the analysis of the salt compounds clear and simple. Specifically, the degree of crystallinity can be determined or confirmed by using, for example, the powder X-ray diffraction (reflection) method described in the following examples, or by using, for example, the powder X-ray diffraction (transmission) method described in the following examples (both are also abbreviated as PXRD below). For crystalline solids having the same chemical composition, different resulting crystal gratings are summarized by the term "polymorphism".

[0227] Preferably, the salt of the present invention, when measured using the PXRD method described herein, exhibits a crystallinity of greater than 30%, more preferably greater than 40%, and even more preferably greater than 50% (e.g., at least 55% to 60%).

[0228] The salts of the present invention can exist as solvates and / or hydrates, which can be formed by the attraction, association, adsorption, adhesion, embedding, or complexation of solvent molecules in the crystal grating of the salts of the present invention. The solvent molecules that can be embedded in the crystal grating can be derived from the solvent used for crystallization and from water with relative humidity.

[0229] The extent to which the selected solvent or water leads to a solvate or hydrate during the process steps or crystallization steps depends on the combination of process conditions and various interactions between the selected compounds (I) or (II) or (II'), the counter anion from the selected acid, and the selected solvent and humidity conditions. A solvate or hydrate of the salt may be preferred because the solvent or water molecules in the crystal structure are bound together by strong intermolecular forces and can therefore represent the structure-forming elements of these crystals, which in part improves the stability of the salt. However, solvent and / or water molecules are also present in some crystal lattices bound together by relatively weak intermolecular forces. Such molecules are more or less integrated into the formation of the crystal structure, but with lower energy effects. The solvent and / or water content of the solvate also depends on drying and environmental conditions (i.e., relative humidity). In the case of stable solvates or hydrates, there is usually a well-defined stoichiometric ratio between the active compound (i.e., the salt) and the solvent or water. In many cases, these ratios do not perfectly meet the stoichiometric values ​​and are often closer to lower values ​​than theoretical values ​​due to certain crystal defects. For weakly bound water, the ratio of organic molecules to solvent or water molecules can vary considerably, extending, for example, beyond dihydrates, trihydrates, or tetrahydrates. On the other hand, in amorphous solids, the molecular structure classification of solvents and / or water is not stoichiometric; however, such classification may also be merely accidental stoichiometry. In some cases, precise stoichiometry of solvent or water molecules is impossible because the formation of layered structures makes it impossible to determine the embedded solvent or water molecules in a definite form.

[0230] The solvent and / or water content in amorphous solids and crystalline solvates or hydrates can usually be determined by conventional methods, such as by using the well-known Karl Fischer titration, by dynamic vapor adsorption (DVS) measurements, by thermogravimetric analysis (TG-FTIR), as described in the examples below. Furthermore, elemental analysis or structural analysis methods, such as... 1 1H NMR spectroscopy or Raman spectroscopy (FT Raman spectroscopy) can provide information about the extent of solvation or hydrate formation, and / or can be used to confirm or verify the results of Karl Fischer (KF), DVS, or TG-FTIR measurements.

[0231] Examples of solvates and / or hydrates according to the invention include, for example, solvates or hydrates with solubilities of half (0.5), one, sesquipartate (1.5), two, three, four, five, six, seven, eight, nine, ten, etc. Further intermediate degrees of solubilization are also possible, for example, solubilizations with solvent and / or water molecules of 2.5, 3.5, 4.5, etc.

[0232] Preferred examples of solvates and / or hydrates include solvates / hydrates having about 0.5, 1, 1.5, 2.5, 3, 4, and 7 solvent / water molecules. Further preferred examples of solvates and / or hydrates include solvates / hydrates having about 0.5, 1, 1.5, 2.5, 3, 4, 6, and 7 solvent / water molecules. More preferably are half- and single-solvates / hydrates having about 0.5 or 1 solvent / water molecule, wherein half-hydrates and single-hydrates are particularly preferred. Anhydrous salts are also preferred. It is also possible that solvent and / or water residues are retained in the salt in non-stoichiometric amounts.

[0233] Furthermore, it is possible that the mixture of water and solvent is retained in the form of a salt, thus forming a so-called mixed hydrate / solvent form. Examples of such mixed hydrate / solvent forms particularly include: acetone / water, preferably in a ratio of 1 to 4:1, for example, especially 4:1; methanol / water, preferably in a ratio of 3 to 9:1, for example, especially 3:1, 4:1 and 9:1; ethanol / water, preferably in a ratio of 1 to 4:1, for example, especially 3:1 and 4:1.

[0234] Any reference in the foregoing and hereinafter to salts of compounds of formula (I) or (II) or (II') should be understood to also refer to the corresponding solvates, such as hydrates, solvates and mixed hydrate / solvates, as well as polymorphic variants and amorphous forms, as applicable.

[0235] Compared to methods known in the prior art, the novel method of the present invention further and surprisingly leads to an increase in yield.

[0236] It is now possible to obtain yields of ≥30%, preferably ≥35%, more preferably ≥40%, and even more preferably ≥45%.

[0237] Conversely, prior art methods provide yields of no more than 22%. For example, the preparation of the 3HCl salt of compound (II) or (II') according to the method of the present invention provides yields of >60%, as shown in Example 4 below. In contrast, the preparation of the 3HCl using the methods described in WO2017 / 068089 and WO2017 / 068090 provides yields of only 22%, as shown in the example of preparing compound 127 in Example 1.

[0238] In particular, the novel condensation method steps of the present invention further provide a more feasible, cost-effective, and labor-efficient approach. Polymer formation during the intermediate formation step can be avoided, which has a positive impact on method control and yield.

[0239] Another aspect of the invention relates to compounds of formula (I) or (II) or (II') as described anywhere herein, including salts, hydrates, solvates, and mixed hydrate / solvate forms, polymorphic variants, and amorphous forms obtainable by the methods described herein. Compounds obtainable by the novel methods described herein are characterized by improved and / or (more) higher purity, defined by a total impurity content of less than 2.00% relative area, preferably less than 1.50% relative area, and more preferably less than 1.00% relative area, wherein the impurity content is determined by HPLC as described in the following examples, and “% relative area” represents the sum of the relative areas of all impurities in the HPLC spectrum.

[0240] Therefore, another aspect of the invention relates to compounds of formulas (I) and (II) or (II') as described anywhere herein, including salts, hydrates, solvates and mixed hydrate / solvate forms, polymorphic variants and amorphous forms, having a total purity of at least 97.80% relative area, preferably at least 97.90% relative area, at least 98.00% relative area, at least 98.10% relative area, at least 98.20% relative area, at least 98.30% area, at least 98.40% relative area, at least 98.50% relative area, at least 98.60% relative area, at least 98.70% relative area, at least 98.80% relative area, at least 98.90% relative area, at least 9 9.00% relative area, at least 99.10% relative area, at least 99.20% relative area, at least 99.30% relative area, at least 99.40% relative area, at least 99.50% relative area, at least 99.60% relative area, at least 99.70% relative area, at least 99.80% relative area, at least 99.90% relative area, at least 99.95% relative area, at least 99.96% relative area, at least 99.97% relative area, at least 99.98% relative area, at least 99.99% relative area, wherein purity is determined by HPLC as described in the following examples, and “% relative area” represents the relative area of ​​the compound of the present invention in the HPLC spectrum.

[0241] In particular, compounds of formulas (I) and (II) or (II') as described anywhere herein, including salts, hydrates, solvates and mixed hydrate / solvate forms, polymorphic variants and amorphous forms, are characterized by containing one or more impurities at relative retention times (RRT) of 0.59, 0.65, 0.83 and 1.37, wherein the amount of these impurities does not exceed 0.20% of the relative area, preferably not more than 0.15% of the relative area, more preferably not more than 0.10% of the relative area, and preferably has a lower limit of 0.05% of the relative area.

[0242] More preferably, compounds of formulas (I) and (II) or (II') as described anywhere herein, including salts, hydrates, solvates and mixed hydrate / solvate forms, polymorphic variants and amorphous forms, are characterized by the absence of impurities at relative retention times (RRTs) of 0.59, 0.65, 0.83 and 1.37.

[0243] More particularly, compounds of formulas (I) and (II) or (II') as described anywhere herein, including salts, hydrates, solvates and mixed hydrate / solvate forms, polymorphic variants and amorphous forms, are characterized by the absence of impurities at relative retention times (RRTs) of 0.27, 0.52, 0.59, 0.65, 0.83, 0.94, 1.19, 1.37, and preferably have a lower limit of 0.05% relative area.

[0244] More particularly, compounds of formulas (I) and (II) or (II') as described anywhere herein, including salts, hydrates, solvates and mixed hydrate / solvate forms, polymorphic variants and amorphous forms, are characterized by an impurity distribution including one or more impurities among impurities at relative retention times (RRTs) of 0.48, 0.70, 1.27 and 1.48.

[0245] Impurities and their retention times (RRTs) were determined by HPLC, as described in the following examples.

[0246] Conversely, the method for preparing compound (II) in the form of a 3HCl salt as described in WO2017 / 068089 and WO2017 / 068090 (preparation of compound 127 in Example) cannot achieve such high purity and improved impurity distribution. Figure 6 , Figure 7 and Figure 8 This can be seen from the text.

[0247] A particularly preferred embodiment of the present invention relates to a polymorph (PM1) of a triHCl salt of compound (II), characterized by a powder X-ray diffraction (PXRD) pattern comprising a characteristic crystalline peak (main peak) expressed in 2-θ degrees at approximately 3.9 degrees and 16.5 degrees ± 0.25 degrees, or ± 0.20 degrees, or ± 0.10 degrees, or ± 0.05 degrees.

[0248] Preferably, in this embodiment of the polymorph (PM1) of the triHCl salt of compound (II), the PXRD spectrum includes one or more other characteristic (main) peaks expressed in 2-θ degrees at about 7.9 degrees, 24.1 degrees, 19.1 degrees, 12.1 degrees and / or 10.0 degrees ± 0.25 degrees or ± 0.20 degrees or ± 0.10 degrees or ± 0.05 degrees.

[0249] More preferably, in this embodiment of the polymorph, the PXRD pattern includes characteristic crystalline (main) peaks expressed in 2-θ degrees at 3.9 degrees, 16.5 degrees, 7.9 degrees, 24.1 degrees, 19.1 degrees, 12.1 degrees, and 10.0 degrees ± 0.20 degrees or 0.10 degrees or 0.05 degrees.

[0250] Preferably, the polymorph (PM1) of the triHCl salt of compound (II) exists in the form of a hemihydrate. Preferably, the polymorph (PM1) of the triHCl salt of compound (II) is characterized by a water activity ≤0.5%, preferably ≤0.4%, more preferably ≤0.3%.

[0251] The melting point (by DSC as described in the examples below) of the polymorph (PM1) of the triHCl salt of compound (II) is preferably in the range of ≥180°C to ≤220°C, more preferably in the range of ≥185°C to ≤215°C, and more preferably in the range of ≥190°C to ≤210°C.

[0252] More preferably, the polymorph (PM1) of the triHCl salt of compound (II) exhibits microcrystallineity (measured by optical microscopy using polarized light), characterized by spherical polycrystalline particles with low crystallinity. The average particle size, measured by optical microscopy using polarized light, is about 10 μm to 50 μm.

[0253] Another characteristic of the polymorph of the triHCl salt of compound (II) (PM1) is its highly fluid nature, which exists as a flowing powder with low electrostatic charge.

[0254] The polymorph (PM1) of the triHCl salt of compound (II) is another thermodynamically stable polymorph at room temperature (23°C ± 5°C).

[0255] Another preferred embodiment of the invention relates to a polymorph (PM2) of the triHCl salt of compound (II), characterized by a powder X-ray diffraction (PXRD) pattern including characteristic crystalline (main) peaks expressed in 2-θ degrees at approximately 16.9 degrees and 25.3 degrees ±0.25 degrees, or ±0.20 degrees, or ±0.10 degrees, or ±0.05 degrees.

[0256] Preferably, in this embodiment of the polymorph (PM2) of the triHCl salt of compound (II), the PXRD spectrum includes one or more other characteristic (main) peaks expressed in 2-θ degrees at about 11.7 degrees, 28.3 degrees, 25.5 degrees, 20.1 degrees and / or 26.2 degrees ±0.25 degrees or ±0.20 degrees or ±0.10 degrees or ±0.05 degrees.

[0257] More preferably, in this embodiment of the polymorph (PM2) of the triHCl salt of compound (II), the PXRD spectrum includes characteristic crystalline (main) peaks expressed in 2-θ degrees at 16.9 degrees, 25.3 degrees, 11.7 degrees, 28.3 degrees, 25.5 degrees, 20.1 degrees and 26.2 degrees ± 0.20 degrees or 0.10 degrees or 0.05 degrees.

[0258] Preferably, the polymorph (PM2) of the triHCl salt of compound (II) is present in an anhydrous form. Preferably, the polymorph (PM2) of the triHCl salt of compound (II) is characterized by a water activity ≤0.8%, preferably ≤0.7%, more preferably ≤0.6%.

[0259] The melting point (by DSC as described in the examples below) of the polymorph (PM2) of the triHCl salt of compound (II) is preferably in the range of ≥210°C to ≤240°C, more preferably in the range of ≥215°C to ≤235°C, and more preferably in the range of ≥220°C to ≤230°C.

[0260] More preferably, the polymorph (PM2) of the triHCl salt of compound (II) exhibits microcrystallineity (measured by using an optical microscope with polarized light), characterized by fine aggregates of needle-like structures with high crystallinity.

[0261] The polymorph of the triHCl salt of compound (II) (PM2) is further characterized by having wool-like solid properties.

[0262] The polymorph (PM2) of the triHCl salt of compound (II) has lower thermodynamic stability at room temperature (23℃±5℃) than that of polymorphs PM1 or PM3.

[0263] A further preferred embodiment of the invention relates to a polymorph (PM3) of a triHCl salt of compound (II), characterized by a powder X-ray diffraction (PXRD) pattern comprising a characteristic crystalline (main) peak expressed in 2-θ degrees at approximately 14.7 degrees and 10.3 degrees ± 0.25 degrees, or ± 0.20 degrees, or ± 0.10 degrees, or ± 0.05 degrees.

[0264] Preferably, in this embodiment of the polymorph (PM3) of the triHCl salt of compound (II), the PXRD spectrum includes one or more other characteristic (main) peaks expressed in 2-θ degrees at about 17.0 degrees, 26.5 degrees, 18.1 degrees, 22.1 degrees and / or 27.1 degrees ±0.25 degrees or ±0.20 degrees or ±0.10 degrees or ±0.05 degrees.

[0265] More preferably, in this embodiment of the polymorph (PM3) of the triHCl salt of compound (II), the PXRD spectrum includes characteristic crystalline (main) peaks expressed in 2-θ degrees at 14.7 degrees, 10.3 degrees, 17.0 degrees, 26.5 degrees, 18.1 degrees, 22.1 degrees and 27.1 degrees ± 0.20 degrees or 0.10 degrees or 0.05 degrees.

[0266] Preferably, the polymorph (PM3) of the triHCl salt of compound (II) exists as a monohydrate. Preferably, the polymorph (PM3) of the triHCl salt of compound (II) is characterized by a water activity > 0.3%.

[0267] The melting point (by DSC as described in the examples below) of the polymorph (PM3) of the triHCl salt of compound (II) is preferably in the range of ≥150°C to ≤190°C, more preferably in the range of ≥155°C to ≤180°C, and more preferably in the range of ≥160°C to ≤175°C.

[0268] More preferably, the polymorph (PM3) of the triHCl salt of compound (II) exhibits microcrystallineity (measured by using an optical microscope with polarized light), characterized by having fine rod-shaped structures with high crystallinity.

[0269] The polymorph of the triHCl salt of compound (II) (PM3) is further characterized by having a large amount of non-flowing powder properties.

[0270] The polymorph (PM3) of the triHCl salt of compound (II) is thermodynamically stable at room temperature (23°C ± 5°C).

[0271] Of the above polymorphs of the trisalt of compound (II), PM1 is the most preferred, particularly because of its thermodynamic stability at room temperature, spherical particulate form, and good flow properties with low electrostatic charge, which are advantageous for use as a pharmaceutically active ingredient.

[0272] In the above embodiments, the terms "characteristic peaks" or "main peaks" refer to those peaks with the highest intensity in the PXRD spectrum. The intensity of the peaks in the PXRD spectrum decreases in the order of the peaks listed above, and the polymorphs (PM1, PM2, and PM3) are preferably characterized by having two or more of those characteristic (main) peaks with the highest intensity.

[0273] The above-mentioned compound (II') has not been disclosed in the prior art, for example in WO2017 / 068089, WO2017 / 068090, WO2018 / 192973 or WO2011 / 029832, and the compound itself is a new compound.

[0274] The compounds of formulas (I), (II), and (II') described herein are particularly suitable for use as pharmaceuticals, acting as inhibitors of membrane siderotransferrins. Inhibition of the membrane siderotransferrins may be determined as described in any one of international patent applications WO2018 / 192973, WO2017 / 068089, and WO2017 / 068090.

[0275] Compounds of formulas (I), (II), and (II') as described herein, including their preferred salts and polymorphs, are particularly suitable for the prevention and / or treatment of iron metabolism disorders leading to increased iron levels or increased iron absorption, such as for the prevention and / or treatment of iron overload and / or for the prevention and / or treatment of diseases associated with or caused by increased iron levels, increased iron absorption, or iron overload. Diseases associated with or caused by increased iron levels, increased iron absorption, or iron overload include, for example, thalassemia, hemoglobinopathies, hemoglobin E disease, hemoglobin H disease, hemochromatosis, hemolytic anemia, thalassemia (including α-thalassemia, β-thalassemia, and δ-thalassemia), sickle cell anemia (sickle cell disease), and congenital erythropoiesis disorders.

[0276] The compounds of formulas (I), (II), and (II') described herein, including their preferred salts and polymorphs, are further suitable for the prevention and / or treatment of diseases associated with ineffective erythropoiesis, such as myelodysplastic syndromes (MDS), polycythemia vera, and congenital erythropoiesis-disordered anemia; or for adjunctive treatment of infections caused by pathogenic microorganisms by limiting the amount of iron available to pathogenic microorganisms (e.g., Vibrio vulnificus); or for the prevention and / or treatment of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases by limiting the deposition or increase of iron in tissues or cells; or for the prevention and / or treatment of the formation of free radicals, reactive oxygen species (ROS), and oxidative stress; or for the prevention and / or treatment of cardiac, hepatic, and endocrine damage caused by iron overload; or for the prevention and / or treatment of inflammation caused by excessive iron.

[0277] Therefore, the present invention further relates to medicaments comprising one or more of compounds (I) or (II) or (II') as defined herein (including their salts, hydrates, solvates, and mixed hydrate / solvate forms, polymorphic variants, and amorphous forms), such as medicaments for the prevention or treatment of any of the aforementioned diseases, conditions, or symptoms. Such medicaments may optionally further comprise one or more pharmaceutical carriers and / or adjuvants and / or solvents, and / or at least one additional pharmaceutically active compound, such as active compounds for the prevention and treatment of iron overload, thalassemia, hemochromatosis or sickle cell disease, neurodegenerative diseases (e.g., Alzheimer's disease or Parkinson's disease), and related symptoms, or iron chelating compounds.

[0278] The drugs described above can be in the form of preparations for oral or parenteral administration.

[0279] Compounds (I) or (II) or (II') as defined herein, including their salts, hydrates, solvates, and mixed hydrate / solvate forms, polymorphic variants, and amorphous forms, are further suitable for use in combination therapies, including co-administration of the compounds of the present invention with at least one additional pharmaceutically active compound, wherein co-administration of the combination therapy can be carried out in a fixed-dose combination therapy by co-administering the compounds of the present invention with at least one additional pharmaceutically active compound in a fixed-dose formulation, or wherein co-administration of the combination therapy can be carried out in a free-dose combination therapy by co-administering the compounds of the present invention and the at least one additional pharmaceutically active compound in free doses of the respective components, or by administering the components simultaneously or by sequentially using the components distributed over a period of time, and wherein the combination therapy preferably includes co-administration of the compounds of the present invention with one or more other pharmaceutically active compounds for reducing iron overload, the one or more other pharmaceutically active compounds being selected from Tmpr ss6-ASO, iron chelating agents, curcumin, SSP-004184, deferitrin, deferasirox, deferoxamine and / or deferoxone, and / or co-administered compounds of the present invention with one or more other pharmaceutically active compounds selected from the following: antioxidants (e.g., n-acetylcysteine), antidiabetic drugs (e.g., GLP-1 receptor agonists), antibiotics (e.g., vancomycin (Van) or tobramycin), drugs for the treatment of malaria, anticancer agents, antifungal drugs, drugs for the treatment of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease (including dopamine agonists such as levodopa), antiviral drugs (e.g., interferon-α or ribavirin), immunosuppressants (e.g., cyclosporine A or cyclosporine A derivatives), iron supplements, vitamin supplements, erythropoiesis stimulants (e.g., erythropoietin, Epo), anti-inflammatory biological products, antithrombolytic agents, statins, vasopressors, and positive inotropic compounds.

[0280] Another aspect of the invention relates to novel intermediate compounds that can be prepared using the novel method steps of the invention, and corresponding methods for preparing them. The method steps for preparing the intermediate compounds may further include crystallization, separation, and / or purification steps for isolating the intermediate compounds.

[0281] Specifically, the present invention relates to intermediate compounds of general formula (IM-2-a).

[0282]

[0283] Another aspect of the invention relates to a method for preparing the intermediate compound (IM-2-a), comprising reaction steps as defined anywhere above in the description of the compound of formula (I), for example, particularly by the following methods:

[0284]

[0285] This may further include crystallization, separation, and / or purification steps for isolating the intermediate compound (IM-1). The preferred method conditions, base, solvent, and catalyst described above are preferably used for the preparation, crystallization, and separation.

[0286] On the other hand, the present invention relates to intermediate compounds of general formula (IM-3).

[0287]

[0288] Among them, X 1 X 2 R 1 R 4 R 5 A and o have the meanings defined anywhere in this document.

[0289] Preferably, the intermediate compound (IM-3) has a group R represented by the following formula. 5

[0290]

[0291] Furthermore,

[0292] o is 1;

[0293] R 1 It is hydrogen;

[0294] A represents the CH group;

[0295] And it is represented by the following formula (IM-3-a).

[0296]

[0297] Among them, X 1 X 2 R 4 and R y It has the meaning as defined anywhere in this document.

[0298] More preferably, in the intermediate compound (IM-3),

[0299] X 1 N represents N;

[0300] X 2 Represents O;

[0301] A represents the CH group;

[0302] o represents 1;

[0303] R 4It represents hydrogen; and

[0304] R y Indicates fluorine or bromine (fluorine being preferred);

[0305] And it is represented by the following formula (IM-3-b) or (IM-3-b'):

[0306]

[0307] Another aspect of the invention relates to a method for preparing intermediate compounds (IM-3), (IM-3-a), (IM-3-b), or (IM-3-b') as defined above, comprising reaction steps as defined anywhere above in the description of the preparation of compounds of formula (I). The preferred method conditions, bases, solvents, and catalysts described above are preferably used for preparation, crystallization, and separation.

[0308] Another aspect of the invention relates to a novel method for preparing intermediate compound (IM-1) or (IM-1-a) as defined anywhere above, the method comprising the reaction steps as described above in the fourth aspect of the invention or in method step 1-a' and more preferably method step 1-a" as described above. The preferred method conditions, base, solvent, and catalyst described above are preferably used for preparation, crystallization, and separation.

[0309] The invention is illustrated in more detail by way of the following examples. These examples are merely illustrative, and those skilled in the art can extend the specific examples to other suitable variations covered below. Attached Figure Description

[0310] Figure 1 PXRD pattern of compound (II) 3HCl salt (polymorph PM1)

[0311] Figure 2 PXRD pattern of compound (II) 3HCl salt (polymorph PM2)

[0312] Figure 3 PXRD pattern of compound (II) 3HCl salt (polymorph PM3)

[0313] Figure 4 PXRD pattern of compound (II) H2SO4 salt

[0314] Figure 5 PXRD pattern of the salt of compound (II) 1H3PO4

[0315] Figure 6HPLC chromatogram showing the impurity distribution of compound (II) 3HCl salt (polymorph PM1) prepared according to the method of Example 4 and subsequently subjected to solvent extraction (method variant 1).

[0316] Figure 7 HPLC chromatogram showing the impurity distribution of compound (II) 3HCl salt (polymorph PM1) prepared according to the method of Example 4 and subsequently subjected to oil separation (method variant 2).

[0317] Figure 8 HPLC chromatogram showing the impurity distribution of compound (II) 3HCl salt (polymorph PM1) obtained by the method described in WO2017068090A1 (preparation of compound 127 in Example).

[0318] Example

[0319] abbreviations

[0320] DCM dichloromethane

[0321] DSC Differential Scanning Calorimetry

[0322] IPC

[0323] HPLC (High-Performance Liquid Chromatography)

[0324] PXRD Powder X-ray Diffraction

[0325] THF Tetrahydrofuran

[0326] Mw molecular weight

[0327] 1a. A method for preparing the intermediate compound (IM-2-a) by crystallization from HCl

[0328] Chemicals:

[0329]

[0330]

[0331] reaction:

[0332]

[0333] process:

[0334] Purge the container with N2 for ≥15 minutes.

[0335] Add LitOBu, add 70 mL THF and stir at 20°C to 25°C for ≥10 min.

[0336] Heat the mixture to 55°C to 60°C and stir for ≥5 minutes.

[0337] Add 15 mL of THF solution of 1,2-dibromoethane at 55°C to 60°C.

[0338] Stir the mixture at this temperature for ≥30 minutes.

[0339] Cool the mixture to 45°C to 50°C.

[0340] Add Pd(PPh3)4, purge with a certain amount of THF and stir at this temperature for ≥5 min.

[0341] The mixture was heated to 60°C to 65°C, and 15 mL of ethyl 4-oxazolyl carboxylate THF solution was added at 60°C to 65°C (exothermic).

[0342] Stir the mixture at this temperature for ≥30 minutes.

[0343] Cool the mixture to 20°C to 25°C.

[0344] Add 35 mL of LiOH aqueous solution at 20°C to 25°C and stir overnight (exothermic).

[0345] IPC by HPLC: ≥95% conversion

[0346] The mixture was extracted three times with isopropyl acetate. The organic phase was then discarded.

[0347] Cool the aqueous phase to 0-5℃ and measure the pH.

[0348] Adjust the pH to 0.9 to 1.1 with 20% HCl while keeping the temperature ≤10℃.

[0349] Stir the suspension at -50℃ for ≥45 min.

[0350] Filter the suspension, wash with 50 mL of cooled (≤5℃) pH 0 HCl and 20 mL of cooled (≤5℃) water, and vacuum dry at 45℃ until dry.

[0351] Yield:

[0352]

[0353] HPLC = 98% area

[0354] q-NMR = 98% m / m

[0355] 1b. Alternative method steps for preparing intermediate compound (IM-1-a)

[0356] In an alternative (but less preferred) method according to Example 1a above, the method steps of stages 1 and 2 may alternatively be carried out starting from compound RM-1, wherein R y It is chlorine and R 4 It is hydrogen, and the compound is referred to herein as RM-1-a', thus yielding the intermediate compound IM-1-a.

[0357] Chemicals:

[0358]

[0359] reaction:

[0360]

[0361] process:

[0362] Ethyl 2-chlorooxazol-4-carboxylate, tributyl(vinyl)tin, and Pd(Ph3P)2Cl2 were added to dioxane under nitrogen atmosphere. The mixture was heated to reflux at 100-110°C for ≥4 h. The mixture was cooled to 20-25°C, filtered through diatomaceous earth, and the filter cake was washed with 200 mL of dioxane. The filtrate was evaporated to dryness under vacuum, and the crude product was purified by chromatography.

[0363] Pillar: Kp-Sil 1500g

[0364] Eluent: EtOAc / Heptane 20:80

[0365] Method: Continuous 7CV, no gradient, threshold 20mAU;

[0366] The crude product was dissolved in EtOAc / heptane 20:80, and two columns were used on this scale.

[0367] Yield:

[0368] Theoretical value Measured value annotation crude product yield - 453.40g Final product yield 146.02g 112.34g 76.93％

[0369] NMR coincidence structure

[0370] 1c. Alternative method steps for preparing the intermediate compound (IM-1-a)

[0371] In an alternative (preferred) method according to Example 1a above, the method steps of stages 1 and 2 may alternatively be carried out starting from compound RM-1, wherein R y It is chlorine and R 4It is hydrogen, and the compound is referred to herein as RM-1-a', thus yielding the intermediate compound IM-1-a.

[0372] reaction:

[0373]

[0374] process:

[0375] Ethyl 2-chlorooxazol-4-carboxylate (RM-1-a' / 1.0 equivalent) was added to RBF, followed by the addition of 2-Me THF (9V) and water (1V) under a nitrogen atmosphere at 25-30°C. Vinylboronic acid pinacol ester (1.2 equivalent) and potassium carbonate (2.5 equivalent) were added to the mixture at 25-30°C, and the resulting mixture was degassed with nitrogen for 15 min. Pd(PPh3)4 (0.05 equivalent) was added under a nitrogen atmosphere, and the reaction mixture was heated to 80°C. The reaction mixture was stirred at 80-85°C for 8-12 h, and the reaction was monitored for completion by TLC / HPLC. After the reaction was complete, the reaction mixture was cooled to 25-30°C and diluted with water (5V). The phases were separated, and the aqueous phase was extracted with 2-Me THF (5V). The combined organic phases were washed with water (5V), followed by washing with a brine solution (5V), and then dried over sodium sulfate. The organic phase was filtered and concentrated under vacuum below 50°C to obtain a crude product as a brown liquid. The crude product was purified by silica gel (60-120 mesh) column chromatography, eluting with ethyl acetate and n-heptane to obtain a pure product.

[0376] NMR coincidence structure.

[0377] Yield:

[0378] 55% to 60%

[0379] purity:

[0380] The purity of the obtained substance is in the range of 96% to 99%.

[0381] Further purification using n-heptane crystallization at lower temperatures below 0°C yielded a purity >98%. When the highly purified material was tested using ICP-MS for palladium content, the content was found to be in the range of 10-250 ppm, which was further reduced to below 25 ppm by treatment with a silane-bonded thiol scavenger (a heterogeneous Pd scavenger).

[0382] 2. Method for preparing intermediate compound (IM-2-a) by crystallization from water

[0383] Chemicals:

[0384]

[0385] reaction:

[0386] The starting compound IM-1-a can be prepared according to the method steps “Stage 1” and “Stage 2” of Example 1a or as described in Example 1b (less preferred).

[0387]

[0388] process:

[0389] IM-1-a was added to the reactor and dissolved in THF.

[0390] Cool the solution to 3°C to 7°C.

[0391] A solution containing LiOH (15.76 g, dissolved in 500 mL of water) was added at 3°C ​​to 7°C and kept for ≥15 min (slightly exothermic).

[0392] Stir the mixture at 3°C ​​to 7°C for ≥3 hours.

[0393] IPC ≥ 97% conversion rate via LC / MS.

[0394] The mixture was extracted twice with DCM. The extraction and separation of the DCM phase were carried out with the aqueous phase at 5°C, but active cooling with DCM at room temperature was not used.

[0395] Each phase should be allowed to stand for ≥30 minutes.

[0396] Discard the organic layer.

[0397] Clean the container with 20% HCl and ethanol.

[0398] Add 20% HCl to the aqueous phase at 3°C ​​to 7°C until the pH reaches 0.5 to 1.0 (slightly exothermic; HCl should not wash the container walls). Near the end of the HCl addition during crystallization, increase the stirrer speed from 80 rpm to 300 rpm for good mixing of the suspension.

[0399] Stir the suspension at 3℃ to 7℃ for ≥30 min.

[0400] Filter the suspension and rinse the reactor once with the mother liquor.

[0401] Wash the wet cake with water at 5°C to 10°C and dry it to dryness at 45°C / vacuum < 50 mbar. Transfer of fine suspensions on the filter may leave some residue in the container, but this residue can be easily removed by rinsing once with the mother liquor.

[0402] Yield:

[0403] Theoretical value Measured value annotation crude product yield - Final yield 83g 77g 93% uncorr.

[0404] NMR measurement: 97% m / m

[0405] Corrected yield: 90%

[0406] 3a. A method for preparing the intermediate compound (IM-3-b) by crystallization from water.

[0407] Chemicals:

[0408]

[0409]

[0410] reaction:

[0411]

[0412] process:

[0413] The starting compound IM-2-a can be prepared as described in Example 1 or 2.

[0414] Compound IM-2-a was suspended in DCM under an inert atmosphere, and 4-methylmorpholine was added at -5°C to 0°C.

[0415] Then add ethyl chloroformate, keeping the temperature ≤0℃ (exothermic addition).

[0416] Add compound RM-2-a (ground) in batches while maintaining a temperature ≤0℃.

[0417] Stir the suspension at -5℃ to 0℃ for ≥2h.

[0418] IPC control of LC / MS achieves a conversion rate of ≥93% per area.

[0419] The mixture was heated to 15°C to 25°C and extracted twice with a 10% NaCl solution.

[0420] The organic phase was extracted three times with 450 mL of HCl at 10% w / w.

[0421] Adjust the combined aqueous phases to pH 2 with 30% NaOH, then adjust the pH to 7 to 8 with 5% NaOH, while maintaining the temperature ≤10℃.

[0422] Filter the suspension, wash twice with 400 mL of water, and vacuum dry at 45 °C until dry.

[0423] Yield:

[0424] Theoretical value Measured value annotation crude product yield - Final yield 178g 162g 91% Uncorr

[0425] NMR measurements: 99%

[0426] Corrected yield: 90%

[0427] 3b. Method for preparing intermediate compound (IM-3-b) — via condensation synthesis of IM-2-a

[0428] reaction:

[0429]

[0430] Method variants 1 ：

[0431] Chemicals:

[0432]

[0433] process:

[0434] The starting compound IM-1-a can be prepared as described in Example 1 or 2.

[0435] The starting compound IM-1-a was dissolved in THF. At an IT temperature of 0°C to 5°C, 20 mL of an aqueous solution containing 1.72 g of LiOH was added. The mixture was stirred at 3°C ​​to 7°C for ≥3 h.

[0436] The conversion rate was ≥97% by HPLC-MS with an IPC of ≥97%.

[0437] Adjust the mixture to an IT of 15°C to 20°C, and within this temperature range, add 20% HCl to set the pH to 0.8 to 1.2. Extract the mixture three times with DCM. Distill off 50 mL at an OT of 30°C / 600 mbar, then add 50 mL of DCM and dry the combined organic phases by removing the azeotropic solvent. Repeat this process until the water content is 0.13%, as determined by the Karl Fischer method.

[0438] Fill the solution to a volume of 160 mL with DCM and cool to an IT of -5°C to 0°C. Add 4-methylmorpholine within this temperature range. Add ethyl chloroformate (exothermic) within this temperature range. Add RM-2-a (ground) in portions at IT ≤ 0°C. Stir the mixture at -5°C to 0°C for ≥ 3 h.

[0439] The conversion rate was ≥93% by HPLC-MS with an IPC of ≥93%.

[0440] Extract the mixture twice with a 10% NaCl solution. Discard the aqueous phase. Extract the organic phase three times with 75 mL of 10% HCl. Discard the organic phase. First, adjust the combined aqueous phases to pH 2 with 30% NaOH at IT ≤ 10 °C, then adjust the pH to 7 to 8 with 5% NaOH. After stirring at ≤ 10 °C for ≥ 15 min, filter the suspension and wash twice with 60 mL of water. Dry the filter cake to dryness at OT 45 °C / < 100 mbar.

[0441] Yield:

[0442]

[0443] HPLC purity = 99.8%

[0444] Water = 0.04%

[0445] In experiments where the DCM water content was reduced to 0.04% through azeotropy, the yield was as high as 85%.

[0446] Method variants 2 ：

[0447] Chemicals:

[0448]

[0449]

[0450] process:

[0451] The starting compound IM-1-a can be prepared as described in Example 1 or 2.

[0452] The starting compound IM-1-a was added to the reactor and filled with THF. The solution was cooled to 0°C to 5°C, and LiOH solution was added at IT ≤ 5°C. The solution was stirred for ≥ 60 min at IT 0°C to 5°C.

[0453] IPC by HPLC:

[0454] If IPC IM-1-a < 0.2% a / a, adjust the pH to 0.5 to 1.0 with 20% HCl at 15°C to 20°C.

[0455] The mixture was extracted three times with DCM. The combined organic phases were dried over MgSO4, filtered, and the filter cake was washed with DCM. The DCM was evaporated at OT 32°C to 37°C / 400 mbar. 8.5 equivalents of DMF were added. The THF was evaporated at 32°C to 37°C / 35 mbar. (The distillation endpoint was reached when no more solvent condensation occurred.)

[0456] Water with IPC ≤ 0.2% via KF:

[0457] If the IPC does not meet specifications, add 10 equivalents of DCM and redistill at 32°C to 37°C, then pass through the IPC KF.

[0458] Add 4-methylmorpholine to the DMF solution at an IT range of -5°C to 0°C. Add ethyl chloroformate at an IT range of -5°C to 0°C. Add RM-2-a (ground) at an IT range of -5°C to 0°C. Stir the mixture at an IT range of -5°C to 0°C for ≥5 hours.

[0459] IPC by HPLC:

[0460] If IPC IM-3-a > 85% a / a, add water slowly at IT < 10°C. If necessary, adjust the pH of the reaction mixture to 6 to 8 with 30% NaOH. Stir the product suspension at IT 0 to 5°C for ≥60 min, filter, wash twice with water, and vacuum dry at 45°C.

[0461] Yield:

[0462] Yield = 448.42g = 75.6%

[0463] HPLC determination = 98.2%

[0464] HPLC purity = 99.7%

[0465] 3c. Method for preparing intermediate compound (IM-3-b) — via condensation synthesis of IM-2-a

[0466] reaction:

[0467]

[0468] Method variant 1 ：

[0469] Chemicals:

[0470]

[0471]

[0472] process:

[0473] Purge with N2 for ≥15 min. Place LitOBu in a container, add 70 mL of THF, and stir the mixture at IT of 20°C to 25°C for ≥10 min.

[0474] Heat the mixture to 53°C to 57°C and stir for ≥5 minutes.

[0475] Add 15 mL of 1,2-dibromoethane THF solution (exothermic) at an IT of 55°C to 60°C, and stir the mixture at an IT of 58°C to 62°C for ≥30 min.

[0476] Cool the mixture to IT at 43°C to 47°C.

[0477] Add Pd(PPh3)4 and stir for ≥5 min.

[0478] Heat the mixture to an IT of 58°C to 62°C, and add 15 mL of ethyl 4-oxazolyl carboxylate THF solution at an IT of 60°C to 65°C.

[0479] Stir the mixture at an IT temperature of 58°C to 62°C for ≥30 min.

[0480] Information obtained by IPC via HPLC:

[0481] -No 4-oxazolylcarboxylate

[0482] - Ethyl ester product 72%

[0483] - tert-butyl ester product 23%

[0484] -Intermediate product IM-1-a 5%

[0485] Cool the mixture to IT at 20°C to 25°C.

[0486] Add 35 mL of LiOH aqueous solution at an IT of 20°C to 27°C, and stir the mixture at an IT of 23°C to 27°C for ≥16 h.

[0487] IPC ≥ 93% by HPLC.

[0488] Add 35 mL of water and extract the mixture twice with 70 mL of TBME.

[0489] Add 50 mL of THF and adjust the pH to 0.5 to 1.0 with 20% HCl at an IT temperature of 15°C to 20°C.

[0490] The mixture was extracted three times with 50 mL of DCM.

[0491] The combined organic phases were dried with Na2SO4 (15 g to 20 g), filtered, and the filter cake was washed with 10 mL of DCM.

[0492] The water content of the DCM phase was determined to be 0.51% by the Karl Fischer method.

[0493] Set OT to -20℃.

[0494] 4-Methylmorpholine was added at an IT temperature of -5°C to 0°C.

[0495] Ethyl chloroformate was added at an IT temperature of -5°C to 0°C.

[0496] Add RM-2-a (ground) in batches at IT≤0℃ and stir the mixture at IT from -5℃ to 0℃ for ≥3h.

[0497] IPC ≥ 90% by HPLC.

[0498] Extract the mixture twice with 100 mL of 10% NaCl.

[0499] The intermediate layer remains together with the organic phase.

[0500] The organic phase was extracted three times with 80 mL of 10% HCl.

[0501] Filter the aqueous solution, first adjust the pH to 2 to 5 with 30% NaOH, then adjust the pH to 7 to 8 with 5% NaOH. Keep the temperature ≤10°C during the pH adjustment process.

[0502] Filter the suspension and wash twice with 60 mL of water.

[0503] The product was dried to dryness at 45°C and <100 mbar.

[0504] Appearance:

[0505] HPLC purity: 99.0%

[0506] Yield:

[0507]

[0508] HPLC purity = 99.8%

[0509] Water = 0.04%

[0510] In experiments where the DCM water content was reduced to 0.04% through azeotropy, the yield was as high as 85%.

[0511] Method variants 2 ：

[0512] Chemicals:

[0513]

[0514]

[0515] process:

[0516] The reactor was purged with nitrogen for 15 minutes, and the condenser was cooled to -30°C.

[0517] Add tBuOLi to the reactor and add THF. Heat the mixture to TI = 55°C (TM = 58°C) and stir for 5 min. Add a THF solution of 1,2-dibromomethane at TI ≤ 60°C. Stir the mixture at TI = 60°C (TM = 63°C) for 60 min. Cool the mixture to TI = 45°C.

[0518] Add Pd(PPh3)4. Heat the mixture to TI = 60℃ (TM = 63℃).

[0519] Add a THF solution of ethyl 4-oxazolyl carboxylate at a temperature of TI ≤ 65°C. Stir the mixture for 30 min at TI = 60°C (TM = 63°C). Cool the mixture to TI = 20°C to 25°C (TM = 20°C).

[0520] IPC by HPLC-MS.

[0521] Add an aqueous solution of LiOH at a temperature of TI ≤ 25°C. Stir the mixture at TI = 25°C for ≥ 16 h.

[0522] IPC by HPLC-MS

[0523] Add water, and extract the mixture twice with MTBE. Discard the MTBE phase. Add THF and adjust the pH to 0.5 to 1.0 with 20% HCl at 15°C to 20°C.

[0524] The mixture was extracted three times with DCM. The combined organic extracts were dried over MgSO4, filtered, and the filter cake was washed with DCM. The DCM was evaporated at 35°C / 400 mbar. 8.5 equivalents of DMF were added, and THF was evaporated at 35°C / 35 mbar. The distillation endpoint was reached when no more solvent condensation occurred.

[0525] For KF, IPC ≤ 0.2% water, otherwise additional azeotropic.

[0526] To further azeotropically distill, 10 equivalents of DCM were added and evaporated at 35°C / 400 mbar. IPC was then repeated via KF.

[0527] Repeat this process until the IPC of KF meets specifications. Add 4-methylmorpholine to the DMF solution at TI = -5℃ to 0℃. Add ethyl chloroformate at TI = -5℃ to 0℃. Add RM-2-a (ground) at TI = -5℃ to 0℃. Stir the mixture at TI = -3℃ for ≥5 hours.

[0528] IPC by HPLC-MS

[0529] If IPC IM2 > 85% a / a, add water when IT < 10°C.

[0530] If necessary, adjust the pH of the reaction mixture to 6 to 8 using 30% NaOH. Stir the product suspension at 0 to 5°C for ≥60 min, filter, wash twice with water, and vacuum dry at 45°C.

[0531] Yield:

[0532] Yield = 5.5g = 62.4%

[0533] HPLC determination = 99.6% m / m

[0534] HPLC purity = 99.7% a / a

[0535] 4. Method for preparing the 3HCl-salt of compound (II)—solvent-exchange condensation synthesis

[0536] Method variant 1 (solvent extraction):

[0537] Chemicals:

[0538]

[0539]

[0540] reaction:

[0541]

[0542] process:

[0543] The intermediate compound IM-3-b can be prepared as described in Example 3.

[0544] Compound RM-3-a (ground) was added to the reactor and suspended in water at 20°C to 25°C.

[0545] Add 30% NaOH and stir the suspension at 20°C to 25°C until a solution is formed (approximately 15 minutes).

[0546] Add intermediate compound IM-3-b (ground) and heat the mixture to 60°C to 65°C for 72 hours.

[0547] IPC conversion rate ≥ 93% of area via LC / MS.

[0548] Cool the mixture to 20°C to 25°C and add DCM.

[0549] Adjust the pH to 3.9 to 4.1 using 10% HCl.

[0550] Stir the mixture for 5 minutes and allow each phase to stand for 1 hour. Discard the lower phase. Repeat DCM extraction three times.

[0551] Adjust the pH of the aqueous phase to 9.9 to 10.1 using 15% NaOH, and add EtOAc.

[0552] Stir the mixture for 5 minutes and allow each phase to stand for 1 hour. Discard the lower phase. Repeat EtOAc extraction twice.

[0553] The organic phases (volume = 8.5 L) were combined and concentrated to a volume of 1.7 L (1 / 5 of the volume) under vacuum / OT at 40 °C.

[0554] Add 3.2 L of EtOH and concentrate the solution to 1.7 L (half its original volume) under vacuum / OT at 40 °C.

[0555] Add 3.2 L of EtOH and concentrate the solution to 1.05 L (1 / 3 of the volume) under vacuum / OT at 40 °C.

[0556] Add 4.55 L of EtOH (5.6 L - 1.05 L), filter the solution and heat to 55°C to 60°C.

[0557] Add 32% HCl within ≥20 min at 55℃ to 60℃, and slowly cool the suspension to 0℃ to 5℃ within ≥3 h.

[0558] Stir the suspension at 0℃ to 5℃ for ≥1h and then filter.

[0559] The filter cake was washed with 0.8 L of EtOH and dried to dryness at 45 °C / vacuum < 50 mbar.

[0560] Yield:

[0561]

[0562] HPLC determination: 75.8% m / m free base, 96.2% m / m salt

[0563] HPLC purity: 99.2% Area

[0564] HPLC purity distribution as follows Figure 6 As shown

[0565] Water content: 1.2%

[0566] Chloride content (elemental analysis): average 19.8% (m / m), which is close to the expected value of 20.5% for a 3:1 HCl:free alkali salt.

[0567] DSC: 192℃

[0568] PXRD analysis: Polymorphic form PM1 (based on Figure 1 (PXRD pattern)

[0569] Method variant 2 (oil separation):

[0570] reaction:

[0571]

[0572] Chemicals:

[0573]

[0574] process:

[0575] RM-3-a was added to the reactor and suspended in water. 30% NaOH was added. IM-3-b was added, and the mixture was heated to IT at 58°C to 62°C for ≥72 h. The IPC of the product (≥75% area, all compounds integrated) was measured by LC / MS.

[0576] Cool the mixture to 5°C to 25°C and allow it to stand for ≥16 h. Separate the bottom oil phase by draining from the reactor, decantation, or by vacuum aspiration of the aqueous phase. Dissolve the separated oil phase in 560 mL of EtOH. Filter the solution and heat to an IT of 60°C to 65°C. Add 0.7 mg of seed crystals. Add 32% HCl over 20 min at an IT of 60°C to 65°C with stirring at 100 rpm. After adding HCl, reduce the stirrer speed to 60 rpm and slowly cool the suspension to an IT of 0°C to 5°C over ≥4 h. Stir the suspension at 0°C to 5°C for ≥1 h and filter. Wash the filter cake with 105 mL of EtOH and dry to dryness at 45°C / vacuum <50 mbar.

[0577] Yield:

[0578] Theoretical value Measured value annotation crude product yield - Final product yield 20.93g 13.7g 65.40％

[0579] HPLC determination: 77.4% m / m free base, 98.3% m / m salt

[0580] Corrected yield = 64.3%

[0581] HPLC purity: 99.7% Area

[0582] HPLC purity distribution as follows Figure 7 As shown

[0583] Preparation of polymorph PM2 in the form of compound (II) 3HCl salt

[0584] The polymorph form PM2 was obtained by thermal recrystallization of the polymorph PM1 of compound (II) 3HCl salt obtained in Example 4 above.

[0585] Thermal recrystallization was carried out in a 1:1 toluene:methanol solvent mixture.

[0586] Polymorphous PM2 1 The 1H NMR spectrum was essentially unchanged compared to that of polymorph PM1, but a distinct singlet at approximately δ3.16 ppm was observed, at which methanol was attributed, indicating a methanol content of approximately 2 mol%.

[0587] Elemental analysis of polymorphic PM2 to determine the amount of chloride provided an average value of 20.0% (m / m), which is very consistent with the expected value of 20.5% for 3:1 HCl: free alkali salt.

[0588] DSC: 226℃

[0589] PXRD analysis: Polymorphic form of PM2 (based on Figure 2 (PXRD pattern)

[0590] Preparation of polymorph PM3 in the form of compound (II) 3HCl salt

[0591] The polymorphic form of PM3 was obtained by the following process: a saturated solution of the polymorphic PM1 of compound (II) 3HCl salt obtained in Example 4 above was prepared in a solvent mixture of acetone and water in a ratio of 9:1 (v / v) at 50°C, cooled to 5°C, and then precipitated as a white solid by adding acetone.

[0592] Polymorphic PM3 1 The H NMR spectrum shows only minor differences compared to that of polymorph PM1, with a slight shift in the broad resonance.

[0593] Elemental analysis of polymorphic PM3 to determine the amount of chloride provided an average of 19.3% (m / m), which is very consistent with the expected value of 20.5% for 3:1 HCl: free alkali salt.

[0594] DSC: 169℃

[0595] PXRD analysis: Polymorphic form of PM3 (based on Figure 3 (PXRD pattern)

[0596] 5. Method for preparing the HCl salt (single salt) of compound (II)—condensation synthesis

[0597] Chemicals:

[0598]

[0599] reaction:

[0600]

[0601] process:

[0602] The intermediate compound IM-3-b can be prepared as described in Example 3.

[0603] The compound RM-3-a (ground) was suspended in water at 20°C to 25°C, and 30% NaOH was added.

[0604] Stir the suspension at 20°C to 25°C for ≥15 minutes to form a solution.

[0605] Add intermediate compound IM-3-b (ground) and heat the suspension to 60°C to 65°C for ≥72 h.

[0606] Conversion rate ≥93% was controlled by HPLC IPC.

[0607] Cool the emulsion to 20°C to 25°C and add 650 mL of DCM.

[0608] Stir the mixture for ≥10 min and allow each phase to stand for ≥1 h. Discard the aqueous phase.

[0609] Add 650 mL of water to the DCM phase and adjust the pH to 5.4 to 5.6 with 20% HCl.

[0610] Stir the mixture vigorously for ≥10 min, and allow each phase to stand for ≥1 h. Discard the DCM phase.

[0611] The aqueous phase was stirred for ≥16 hours, and the resulting suspension was filtered.

[0612] Wash the wet cake with 80 mL of EtOH.

[0613] The filter cake was dried at 50°C / <100 mbar until dry.

[0614] Yield:

[0615] Theoretical value Measured value annotation Crude product yield - Final product yield 36.0g 21.0g 59,00％

[0616] HPLC determination: 88.7% m / m free base, 96.6% m / m

[0617] Corrected yield = 57%

[0618] HPLC purity = 98.4% Area

[0619] DSC = 173℃

[0620] 6. Method for preparing the H2SO4 salt of compound (II)—condensation synthesis

[0621] Chemicals:

[0622]

[0623] reaction:

[0624]

[0625] process:

[0626] The intermediate compound IM-3-b can be prepared as described in Example 3.

[0627] The compound RM-3-a (ground) was suspended in water at 20°C to 25°C. 30% NaOH was added, and the mixture was stirred at 20°C to 25°C for ≥15 minutes.

[0628] Add intermediate compound IM-3-b (ground) and heat the mixture to 60°C to 65°C for ≥72 h.

[0629] IPC conversion rate via HPLC is ≥93%.

[0630] Cool the mixture to 20°C to 25°C.

[0631] Add 640 mL of DCM, stir, and let each phase stand for ≥1 h. Discard the aqueous phase.

[0632] Add 640 mL of water to the DCM phase and adjust the pH to 3.4 to 3.6 with 20% H2SO4. Let each phase stand for ≥1 h and discard the DCM phase.

[0633] Stir the aqueous phase and cool it to 0°C to 5°C within ≥2 hours.

[0634] The suspension was heated to 40°C and stirred at that temperature for ≥16 hours. The suspension was then cooled to 0°C to 5°C over ≥4 hours and stirred at 0°C to 5°C for ≥1 hour.

[0635] Filter the suspension and wash the filter cake with 160 mL of ethanol.

[0636] The filter cake was dried to dryness at 50°C / <100 mbar vacuum.

[0637] Yield:

[0638]

[0639] HPLC analysis: 76.4% m / m free base, 94.8% m / m 1:1 salt

[0640] Corrected yield = 57%

[0641] HPLC purity: 98.3%

[0642] H2O / KF = 2.3%

[0643] DSC: 176℃

[0644] 7. Method for preparing the 0.5H3PO4 salt of compound (II)—condensation synthesis

[0645] Chemicals:

[0646]

[0647] reaction:

[0648]

[0649] process:

[0650] The intermediate compound IM-3-b can be prepared as described in Example 3.

[0651] The compound RM-3-a (ground) was suspended in water at 20°C to 25°C, and 30% NaOH was added.

[0652] The suspension was stirred at 20°C to 25°C for ≥15 min to form a solution. The intermediate compound IM-3-b (ground) was added, and the suspension was heated to 60°C to 65°C for ≥72 h.

[0653] Conversion rate ≥93% was controlled by HPLC IPC.

[0654] Cool the emulsion to 20°C to 25°C and add 320 mL of DCM.

[0655] Adjust the pH to 3.9 to 4.1 using 20% ​​HCl.

[0656] Stir the mixture vigorously for ≥10 min, and allow each phase to stand for ≥1 h. Discard the organic phase.

[0657] Repeat the DCM extraction three more times.

[0658] Adjust the pH of the aqueous phase to 9.9 to 10.1 using 30% NaOH.

[0659] Add 320 mL of EtOAc and stir the mixture vigorously for ≥10 min. Let each phase stand for ≥1 h. Discard the aqueous phase.

[0660] Repeat the EtOAc extraction twice more.

[0661] The combined organic phases were concentrated to 13 mL under vacuum at 40 °C.

[0662] Add 320 mL of ethanol and concentrate the solution to a volume of 18 mL.

[0663] Add 110 mL of ethanol and 5 × 6.5 mL of 30% H3PO4 at 20°C to 25°C. Stir the mixture at 28°C to 32°C for ≥24 h. Cool the suspension to 20°C to 25°C and filter over ≥1 h. Wash the filter cake with 40 mL of EtOH.

[0664] The filter cake was dried at 45°C and <100 mbar until dry.

[0665] Yield:

[0666] Theoretical value Measured value annotation crude product yield - Final product yield 18.5g 8.8g 48.00％

[0667] HPLC determination: 85.2% m / m free base, 95.4% m / m 2:1 salt

[0668] Corrected yield = 45.8%

[0669] HPLC purity = 99.5% Area

[0670] P = 3.3%

[0671] DSC = 253℃

[0672] 8. Method for preparing the H3PO4 salt of compound (II)—conversion to a 1:1 salt

[0673] Chemicals:

[0674]

[0675] reaction:

[0676]

[0677] process:

[0678] The 0.5H3PO4 salt of compound (II) can be prepared as described in Example 7.

[0679] The salt of compound (II) 0.5H3PO4 was suspended in ethanol at 20°C to 25°C and stirred at this temperature for 4 days.

[0680] Filter the suspension and dry the wet cake at 45°C / <100 mbar until dry.

[0681] Yield:

[0682] Theoretical value Measured value annotation Crude product yield - Final product yield 13.3g 6.6g 50.00％

[0683] Yield: 6.6g = 50%, relative to free alkali

[0684] HPLC analysis: 78.3% m / m free base, 97% m / m 1:1 salt

[0685] Corrected yield = 48.5%

[0686] HPLC purity: 98.7% Area

[0687] P = 5.7%

[0688] DSC: 182℃

[0689] 9a. Method for preparing intermediate compound (IM-3-b')

[0690] Chemicals:

[0691]

[0692] reaction:

[0693]

[0694] process:

[0695] The reaction was carried out under a flow of N2.

[0696] 2-Vinyloxazole-4-carboxylic acid ester (IM-2-a) was suspended in dichloromethane and cooled to -5°C.

[0697] 4-Methylmorpholine was added dropwise at a temperature not exceeding 0°C (exothermic).

[0698] Ethyl chloroformate was added dropwise at a temperature not exceeding 0°C (exothermic).

[0699] After stirring for 20 minutes, 2-(aminomethyl)-3-bromopyridine (RM-2-a') is added at a temperature not exceeding 0°C.

[0700] Stir the mixture overnight at 0°C to 5°C.

[0701] The mixture was washed three times with a 10% NaCl solution.

[0702] The organic phase was concentrated in a rotary evaporator at 45°C.

[0703] Add EtOAc / heptane 20:80 (1.7 mL / g) to the crude product, stir at RT for 4 h, filter, wash twice with EtOAc / heptane and dry at 50 °C to constant weight.

[0704] Yield:

[0705] Theoretical value Measured value annotation Yield 4.76g 4.50g 94.54％

[0706] The theoretical yield was observed to be 94.5% for 4.5g of grayish-brown solid.

[0707] 1 H-NMR corresponds to a theoretical yield of 94.5% for the provided structure.

[0708] 9b. Methods for preparing compound (II')

[0709] Chemicals:

[0710]

[0711]

[0712] reaction:

[0713]

[0714] process:

[0715] Benzimidazole-ethylamine (RM-3) was ground in a mortar and then suspended in water.

[0716] Add 30% sodium hydroxide solution dropwise and stir at RT for 10 min.

[0717] The intermediate compound IM-3-b' was added to the mixture and stirred at an internal temperature of 63°C for 72 hours.

[0718] Cool the mixture to RT.

[0719] Add 110 mL of DCM and stir for 10 minutes.

[0720] Set the pH to 4 using 20% ​​HCl.

[0721] The aqueous phase was extracted four times with 110 mL of DCM, and the phases were separated by standing for 1 hour. The DCM phase was then discarded.

[0722] Adjust the pH of the aqueous phase to 10 with 30% NaOH, extract three times with EtOAc, let stand for 1 hour for phase separation, and discard the aqueous phase.

[0723] The EtOAc phase was dried with magnesium sulfate, filtered, and concentrated to dryness on a rotary evaporator.

[0724] Yield:

[0725] Theoretical value Measured value annotation Yield 6.56g 4.18g 63.72％

[0726] The theoretical yield was observed to be 63.7% for 4.18g of brown oil.

[0727] 1 H-NMR corresponds to the structure

[0728] 9c. Method for preparing the 3HCl salt of compound (II')

[0729] Chemicals:

[0730]

[0731]

[0732] reaction:

[0733]

[0734] process:

[0735] Compound (II') was dissolved in ethanol and heated to an internal temperature of 40°C.

[0736] Add 32% HCl dropwise.

[0737] The suspension was slowly cooled to 0°C.

[0738] The suspension was stirred at 0°C for 1 hour.

[0739] The suspension was filtered, the wet cake was washed with ethanol and dried at 40°C until dry.

[0740] Yield:

[0741] Theoretical value Measured value annotation Yield 5.13g 4.02g 78.36％

[0742] The theoretical yield was observed to be 78.4% for 4.02g of white solid.

[0743] 1 H-NMR corresponds to the structure

[0744] 10. Intermediate compounds and example compounds 1 H-NMR, PXRD and DSC analysis

[0745] 10.1 NMR Analysis

[0746] The intermediate compounds IM-1-a, IM-2-a and IM-3-b prepared by the methods described in Examples 1b, 1a and 3a, respectively, have been subjected to NMR analysis, and the following NMR data are provided.

[0747]

[0748]

[0749] The following NMR data are provided by NMR analysis of the 3HCl salt (polymorphic forms PM1, PM2, and PM2), 1HCl salt, H2SO4 salt, 0.5H3PO4 salt, and 1H3PO4 salt of compound (II) of Examples prepared by the methods described in Examples 4, 5, 6, 7, and 8, respectively.

[0750]

[0751] The following NMR data are provided by NMR analysis of the intermediate compound IM-3-b', compound (II'), and 3HCl salt of compound (II') prepared by the methods described in Examples 9a, 9b, and 9c, respectively.

[0752]

[0753] 10.2 PXRD Analysis

[0754] Further PXRD analysis was performed on the 3HCl salts (PM1, PM2, and PM3), H2SO4 salts, and 1H3PO4 salts of compound (II) of Examples prepared by the methods described in Examples 4, 6, and 8, respectively, to provide, as Figures 1 to 5 The PXRD spectrum shown is shown.

[0755] method

[0756] Sample preparation and measurement:

[0757] Sample preparation: Place 10 mg to 20 mg of sample between two acetic acid foils in the Stoe transmission sample holder; rotate the sample during measurement.

[0758] -Stoe Stadi P (G.52.SYS.S072); Mythen 1K detector; Cu-Ka1 irradiation; Standard measurement conditions: transmission; 40kV and 40mA tube power; Bent Ge monochromator;

[0759] -0.02°2θ step size, 48s step time, 1.5-50.5°2θ scan range; detector mode: step scan; 1°2θ detector step;

[0760] Data evaluation:

[0761] d-value analysis was performed using EVA software (version 14,0,0,0) from Bruker.

[0762] - Subtract background

[0763] - Only lines up to 35°2θ are listed.

[0764] - Calculate relative strength using formulas in Excel

[0765] PXRD conditions—Compound (II) 3HCl salt ( Figure 1 , Figure 2 and Figure 3 PM1, PM2 and PM3)

[0766]

[0767]

[0768] Polymorph PM1:

[0769]

[0770] Polymorphic PM2:

[0771]

[0772]

[0773] Polymorphic PM3:

[0774]

[0775]

[0776] PXRD conditions—Compound (II) H2SO4 salt ( Figure 4 )

[0777]

[0778]

[0779]

[0780]

[0781] PXRD conditions—compound (II)H3PO4 salt ( Figure 5 )

[0782]

[0783]

[0784]

[0785]

[0786] 10.3 DSC Measurement

[0787] Differential scanning calorimetry was performed in a sealed gold disk at a heating rate of 10 °C / min.

[0788] 10.4 HPLC determination of identification, impurity distribution and purity

[0789] Based on the European Pharmacopoeia 2.2.29, using laboratory methods, qualitative and quantitative analysis by high performance liquid chromatography (HPLC) with diode array detection was performed to identify and determine the bases and impurities of the compounds of the present invention.

[0790] The identification of the synthesized compound was confirmed by comparing its retention time and spectrum with that of the corresponding reference material. For the salt form and its impurities, the determination was calculated based on the external calibration curve of the analyte in the base form.

[0791] process

[0792] equipment

[0793] HPLC systems are equipped with pumps, autosamplers, column ovens, diode array detectors, analytical balances (Class 1), pipettes (e.g., Gilson, Rainin), volumetric flasks (10 mL, 25 mL, and 50 mL), and membrane filters (0.2 μm).

[0794] reagents

[0795] - Methanol (HPLC grade), Sigma (or equivalent)

[0796] - For HPLC analysis of 100% trifluoroacetic acid (TFA), Sigma (or equivalent)

[0797] Mobile phase:

[0798] A: Water / Methanol (95+5V / V; 0.1% TFA)

[0799] B: Methanol / Water (95+5V / V; 0.1% TFA)

[0800] Mobile phase A: Water / methanol (95+5V / V; 0.1% TFA)

[0801] Dilute 100 mL of methanol with water R to 2 liters. Then add 2.0 mL of TFA, mix thoroughly, and degas in an ultrasonic bath.

[0802] Mobile phase B: Methanol / water (95+5V / V; 0.1% TFA)

[0803] Dilute 50 mL of water R with methanol to 1 L. Then add 1.0 mL of TFA, mix thoroughly, and degas in an ultrasonic bath.

[0804] Stock solutions A and B (1000 mg / L): Two stock solutions containing 1,000 mg / L of the test compound were prepared by dissolving 50 mg of the test compound in 50 mL of mobile phase A.

[0805] Standard solutions A and B (40 mg / L): Each stock solution was diluted 1:25 with mobile phase A. Samples were filtered into HPLC vials through a 0.2 μm filter.

[0806] SST stock solution

[0807] Weigh 10 mg of a reference impurity, such as the intermediate compound expected in the method, and place it in a 10 mL volumetric flask. Add 5 mL of methanol and dilute with water.

[0808] Impurity solution C (1 mg / L)

[0809] Transfer 100 μL of stock solution A to a 100 mL volumetric flask and dilute with mobile phase A. Filter the sample through a 0.2 μm filter into an HPLC vial.

[0810] Impurity solution D / SST standard solution (10 mg / L)

[0811] Transfer 1000 μL of stock solution B and 1000 μL of SST stock solution to a 100 mL volumetric flask and dilute with mobile phase A. Filter the sample through a 0.2 μm filter into an HPLC vial.

[0812] Chromatography system

[0813] The liquid chromatography system was equipped with a column oven with a controlled temperature of 40℃ ± 5℃, a diode array detector (200nm to 400nm), and a 4.6mm × 150mm C18 column (e.g., Ascentis Express C18, 2.7μm). The flow rate was 1.0mL / min.

[0814] LC gradient:

[0815]

[0816] Runtime: 20.0 min

[0817] UV wavelength: 254nm

[0818] Injection volume: 10 μL for sample, variable volume for standard solution.

[0819] System suitability testing (SST) and quality control checks (QC)

[0820] If the RSD and retention time of the peak area of ​​the analyte are ≤2%, the resolution of the analyte is ≥1.5, and the tailing factor of the analyte peak is in the range of 0.8 to 1.5, then the adequate injection of the SST standard solution proves the suitability of the chromatographic system.

[0821] Standard solution A is injected six times at the beginning of the sequence (SST) and twice at the end of the sequence (QC). Sequencing can only be used if these requirements are met.

[0822] Standard curve

[0823] A standard curve for the analyte (determination) can be obtained, for example, by injecting 5.0 μL, 7.5 μL, and 10.0 μL of standard solution A and 12.5 μL and 15.0 μL of standard solution B.

[0824] A standard curve of impurities of the analyte compound can be obtained, for example, by injecting 5.0 μL and 10.0 μL of impurity solution C and 5.0 μL, 10.0 μL and 15.0 μL of impurity solution D.

[0825] The regression coefficients of the two standard curves should be ≥0.9990.

[0826] Sample preparation

[0827] Process—Impurities

[0828] Accurately weigh approximately 50 mg of the analyte in salt form and place it in a 50 mL flask, then dilute to the mark with mobile phase A. Prepare the sample solution twice. Filter the solution through a 0.2 μm membrane filter and inject 10 μL.

[0829] Process—Measurement

[0830] Two solutions prepared as "impurities" were used and diluted with mobile phase A at a ratio of 1:25. The solutions were filtered through a 0.2 μm membrane filter and injected in 10 μL increments.

[0831] calculate

[0832] Qualitative determination

[0833] The retention time of the analyte should not deviate from the peak in standard solution A by more than 0.3 min. The spectrum of the analyte in the obtained sample should correlate with the reference spectrum of the analyte in standard solution A. Determine the library matching factor. The latter should be ≥990.

[0834] The retention time of any impurity in the analyte should not deviate from the corresponding reference material by more than 0.1 min. The spectra of the impurities in the obtained sample should be correlated with the reference spectra of the corresponding impurities in the SST solution. Determine the library matching factor. The latter should be ≥990.

[0835] Quantitative determination — determination

[0836] Plot the concentration of the standard solution (expressed as the amount of the analyte in its base form (μg)) against the peak area, and extract the intercept (a) and slope (b). The obtained correlation coefficient should not be less than 0.999. Calculate the amount of the analyte base using the following formula:

[0837]

[0838] y Peak area of ​​the sample (analyte compound)

[0839] a. Intercept of the standard curve

[0840] b. Slope of the standard curve (area / μg)

[0841] V1 dilution (e.g., 50 mL)

[0842] V2 is the volume of the sample diluted by a dilution factor (e.g., 1.0 mL / 25 mL = 25).

[0843] V3 Injection volume (e.g., 10 μL)

[0844] E. Sample volume (mg) weighed.

[0845] 100 is the factor [m / m] used to calculate %

[0846] Quantitative determination—impurities

[0847] Quantitative determination of impurities, expressed as % area.

[0848] Integrate all peaks at 254 nm in the sample chromatogram. Peaks related to the blank are not considered. Determine and report the total impurities and the percentage of area for each individual impurity that is ≥0.05%.

[0849] Quantitative determination of impurities, expressed as %m / m

[0850] Plot the concentration of the standard solution (expressed as the amount (μg) of the analyte in its base form) against the peak area, and extract the intercept (a) and slope (b). The obtained correlation coefficient should not be less than 0.999. Considering the response factor, calculate the amount of each impurity using the following formula:

[0851]

[0852] y Peak area of ​​the sample (analyte compound)

[0853] a. Intercept of the standard curve

[0854] b. Slope of the standard curve (area / μg)

[0855] V1 dilution (e.g., 50 mL)

[0856] V2 Injection volume (e.g., 10 μL)

[0857] E. Sample volume (mg) weighed.

[0858] 100 is the factor [m / m] used to calculate %

[0859] RF response factor of each impurity to the analyte compound

[0860] The above method is used to identify and determine the 3HCl salt (polymorph PM1) of compound (II) prepared according to the method of Example 4 (method variants 1 and 2).

[0861]

[0862]

[0863]

[0864] impurities Response factor RM-2-a = RM2 = SP6 1.140 RM-3-a = RM3 1.269 SP1 1.300 SP3 0.916 SP4 0.884 SP5 0.889 SP7 0.748

[0865] result:

[0866]

[0867] Figure 6 The HPLC chromatogram shows the impurity distribution of compound (II) 3HCl salt (polymorph PM1 / VIT-2763) prepared by solvent extraction (method variant 1) according to the method of Example 4.

[0868]

[0869] Figure 7The HPLC chromatogram shows the impurity distribution of compound (II) 3HCl salt (polymorph PM1 / VIT-2763) prepared by subsequent oil separation (method variant 2) according to the method of Example 4.

[0870]

[0871] Comparative Example—Compound 127 according to Example 1 of WO2017068090A1:

[0872] Figure 8 An HPLC chromatogram of the impurity distribution of compound (II) 3HCl salt (polymorph PM1 / VIT-2763) obtained by the method described in WO2017068090A1 (preparation of compound 127 in Example) is shown.

[0873]

[0874] Compound (II) is converted from a base to 3HCl.

[0875] VIT-2763-3HCl [% (m / m)] = VIT-2763 base [% (m / m)] × 517.81 [g / mol] / 408.44 [g / mol]

[0876] This corresponds to a conversion factor of 1.27.

[0877] 11. Pharmacological assays for evaluating the activities of compounds (II) and (II').

[0878] The table below summarizes the activities of compounds (II) and (II') as inhibitors of membrane iron transporters compared to hepcidin.

[0879]

[0880]

[0881] Compounds (II) and (II') were tested in the form of their 3HCl salts.

[0882] 11.1 Determination of internalization of hepcidin (J774)

[0883] This cellular assay allows for the quantification of hepcidin binding to the membrane iron transporter (Fpn) by microscopic detection of the internalization of fluorescently labeled hepcidin into J774 cells. J774 is a mouse macrophage cell line that exhibits endogenous expression of Fpn after co-incubation with iron (Knutson et al., 2005). Binding of hepcidin to Fpn triggers the internalization and degradation of both hepcidin and Fpn. However, the TMR (6-carboxytetramethylrhodamine) fluorophore attached to hepcidin remains associated with the cell even after the degradation of the hepcidin peptide backbone. Therefore, microscopic detection of cell-associated TMR fluorescence is a measure of hepcidin-Fpn binding and the internalization of both hepcidin and Fpn. If TMR-hepcidin-Fpn binding is prevented, cellular TMR fluorescence remains low (Dürrenberger et al., 2013). The role of low molecular weight Fpn inhibitor compounds in this assay was evaluated in vitro as described below.

[0884] J774 cells were harvested from approximately 80% confluence cultures and cultured at 8 × 10⁻⁶ cells / year. 5Cells were seeded at a concentration of 100 μL / mL in complete medium (DMEM, 10% FBS, 1% penicillin-streptomycin) containing 200 μM Fe(III)NTA (triazine) and grown in 96-well MicroClear plates (Greine; catalog 655090) at 37°C and 5% CO2. After overnight incubation, cells were washed three times with pre-warmed DMEM w / o phenol red. After the last wash, 30 μL / well of DMEM w / o phenol red and 10 μL / well of serial test compound dilutions were added, in triplicate. J774 cells were pre-incubated with the test compounds at 37°C and 5% CO2 for 15 min before adding a final concentration of 25 nM TMR-hepcidin. Cells were incubated at 37°C and 5% CO2 for 2 hours, followed by incubation with Hoechst 33342 dye to a final concentration of 0.5 μg / mL to stain the cell nuclei. The cells were then incubated again at 37°C and 5% CO2 for 10 min. Cells were washed three times with PBS and fixed for 15 min at room temperature with 100 μL of 4% paraformaldehyde in PBS. After removing the paraformaldehyde solution, cells were washed three times with PBS, leaving 100 μL per well, and the plates were sealed with foil. TMR (530 nm to 550 nm excitation / 575 nm to 625 nm emission / 400 ms exposure time) and Hoechst 33342 (360 nm to 370 nm excitation / 420 nm to 460 nm emission / 10 ms exposure time) fluorescence images were acquired using a ScanR flatbed imager (Olympus) with a 20x high-NA objective lens. Four images were acquired per well, with the fluorescence channel covering approximately 1500 cells per well. The acquired image data were analyzed using ScanR image analysis software. Image analysis included nucleus detection (Hoechst 33342 fluorescence), identification of cell-related regions, application of virtual channels, and thresholding for rolling ball background reduction. The Sum(Mean) algorithm was then applied to measure TMR fluorescence associated with the cells as a quantitative measure of internalized TMR-hepcidin. IC50 was calculated using the "log(inhibitor) vs. response" curve fitting function of Prism 5 software (GraphPad, version 5.02) with the Sum(Mean) algorithm on the raw data. 50 Values. For each dataset, the fit of the "log(inhibitor) versus response (three parameters)" model is compared with the fit of the "log(inhibitor) versus response - variable slope (four parameters)" model, and the IC of the preferred model is used. 50 Data. Tested in the ferricyanide internalization assay according to formulas (II) and (II'). IC of the compound 50 The data are shown in Table 1. In this determination, the IC50 of unlabeled ferricyanide was...50 It is 0.015μM±0.011μM.

[0885] Table 1 shows the average (AVE) IC50 of compounds (II) and (II') tested in the hepcidin internalization assay. 50 Data used for multiple measurements

[0886] Experimental compounds J774 IC50(μM) Compound (Ⅱ') 0.006 Compound (II) 0.007

[0887] 11.2 Biophysical assay of membrane iron transporter-hepcidin binding

[0888] This biophysical assay was developed to more directly confirm the inhibition of hepcidin binding to membrane iron transporter (Fpn). TMR-hepcidin was incubated with purified human Fpn isolated from Pichia pastoris yeast cells expressing human Fpn with a C-terminal FLAG affinity tag (Bonaccorsi di Patti, 2014), and incubation resulted in an increase in the fluorescence polarization (FP) of the TMR-hepcidin ligand. The inhibition of TMR-hepcidin binding to Fpn by compounds (II) and (II') was tested, as detected by a dose-dependent decrease in the TMR FP signal, as detailed below.

[0889] A mixture of 1.3 mM human Fpn and 30 nM TMR-hepcidin with FP assay buffer (containing 50 mM Tris-HCl (pH 7.3), 200 mM NaCl, 0.02% DDM, and 0.1% BSA) was seeded at 16 mL per well into 384-well black low-volume round-bottom plates (Corning, catalog 3677). Two 8 mL aliquots of serial dilutions of the test compound were added to achieve a final Fpn concentration of 1 mM and a final TMR-hepcidin concentration of 20 nM, respectively. The plates were incubated at room temperature for 90 min, and parallel (S) and vertical (P) fluorescence were measured using a Synergy H1 fluorescence reader (BioTek). FP values ​​were calculated in mP according to the following formula.

[0890]

[0891] IC 50 The values ​​were determined by calculated mP values, as described in the hepcidin internalization assay, and are listed in Table 2. In this assay, the IC50 value of unlabeled hepcidin was... 50 It is 0.37μM±0.067μM.

[0892] Table 2 shows the average (AVE) IC50 of compounds (II) and (II') tested in the biophysical hepcidin-membrane iron transporter binding assay. 50 The data is used for multiple measurements.

[0893] Experimental compounds FP IC50 (μM) Compound (Ⅱ') 0.028 Compound (II) 0.028

[0894] 11.3 Inhibition of membrane iron transporter-mediated iron export activity in iron response assays

[0895] In this assay, intracellular iron levels were indirectly measured by monitoring the activity of a β-lactamase (BLA) reporter gene fused to the human ferritin promoter and the associated iron regulatory elements (IREs) contained in the 5' untranslated region of ferritin mRNA. Expression of the membrane iron transporter (Fpn) in this cell line leads to iron efflux and lower iron levels, reflected in lower reporter gene activity. Conversely, inhibition of Fpn-mediated iron efflux resulted in increased cellular iron levels, which was detected as increased reporter gene activity. The dose-dependent effects of low molecular weight Fpn inhibitory compounds were tested in this in vitro iron response assay, as described below.

[0896] HEK-293 cell line #354 was generated by (i) stably integrating a human Fpn-GFP fusion construct into a derivative of the doxycycline-inducible pTRE-Tight-BI plasmid (Clontech, Cat. 631068) and (ii) a human ferritin promoter-BLA reporter gene into a derivative of the HEK-293Tet-ON Advanced cell line (Clontech). To generate the ferritin-BLA reporter gene construct, a 1.4 kb fragment of the human ferritin H promoter was amplified from human genomic DNA by PCR (forward primer 5'-CAGGTTTGTGAGCATCCTGAA-3'; reverse primer 5'-GGCGGCGACTAAGGAGAGG-3') and inserted into… 6.2 / cGeneBLAzer TM - The BLA gene present in the DEST plasmid (Invitrogen, Cat. 12578-043) was replaced, thus replacing the original CMV promoter and placing the IRE regulating ferritin gene translation approximately 170 bp upstream of the reporter gene start codon. #354 cells were harvested from approximately 80% confluence cultures and cultured at 1.8 × 10⁻⁶. 5 Cells / mL seeded in DMEM / F12 GlutaMAX TMThe culture medium (Invitrogen, Cat. 31331-028) contained 10% FBS (Clontech, Cat. 631106), 1% penicillin-streptomycin, 200 μg / mL hygromycin B (Invitrogen, catalog 10687-010), 5 μg / mL blasticin (Invitrogen, Cat. R210-01), and 4 μg / mL doxycycline (Clontech, Cat. 631311). 50 μL was added to each well of a 384-well PDL-coated plate and grown at 37°C and 5% CO2. After overnight incubation, 10 μL / well of the test compound was added in quadruplicate, and the plate was further incubated overnight at 37°C and 5% CO2. Cells were washed three times with HBSS, leaving 25 μL in each well. BLA activity was detected by adding 5 μL / well of GeneBlazer reagent CCF4-AM (Invitrogen, Cat. K1085) to cells. After incubating the plates in the dark at 18°C ​​for 60 min, blue and green fluorescence signals were measured using a Safire 2 fluorescence microplate reader (Tecan), with excitation at 410 nm and emission at 458 nm (blue) and 522 nm (green). The blue / green fluorescence ratio was calculated as a measure of BLA activity, and EC was determined using the calculated blue / green fluorescence ratio. 50 Values, as described in the hepcidin internalization assay. EC values ​​of the tested compounds (II) and (II'). 50 The data are listed in Table 3. In this determination, the EC of hepcidin was... 50 It is 0.096μM ± 0.063μM (n = 37).

[0897] Table 3 shows the average (AVE) EC of compounds (II) and (II') tested in the iron response assay. 50 The data is used for multiple measurements.

[0898]

[0899] 11.4 Determination of internalization and degradation of membrane iron transporters

[0900] HEK-293 cell line #354 (described in 11.3) was used to measure the ability of compounds to induce the internalization and degradation of membrane iron transporter (Fpn) by fluorescence-activated cell sorting (FACS). Growth of HEK-293#354 cells in a medium containing doxycycline induced the expression of the human Fpn-GFP fusion protein on the cell surface. Data from 10 independent experiments showed that culturing HEK#354 cells at 4 μg / mL doxycycline for 48 h induced an average of 42.6% ± 6.4% of Fpn-GFP positive cells. The dose-dependent effect of low molecular weight Fpn inhibitor compounds on the mean fluorescence intensity (MFI) of Fpn-GFP in HEK-293 cell line #354 was tested, as described below.

[0901] HEK#354 cells were harvested from approximately 80% confluence cultures and cultured at 0.6 × 10⁻⁶ cells per cell line. 6 Cells / mL seeded in DMEM / F12 GlutaMAX TM The culture medium (Invitrogen, Cat. 31331-028) contained 10% FBS (Clontech, Cat. 631106), 1% penicillin-streptomycin (Invitrogen, Cat. 15140-122), 200 μg / mL hygromycin B (Invitrogen, Cat. 10687-010), 5 μg / mL blastomycin (Invitrogen, Cat. R210-01), and 4 μg / mL doxycycline (Clontech, Cat. 631311). 50 μL was added to each well of a 384-well plate (Greiner; Cat. 781091) and the plates were grown at 37°C and 5% CO2. After overnight incubation, 10 μL / well of the test compound was added in quadruplicate, and the plates were further incubated overnight at 37°C and 5% CO2. Cells were washed once with FACS buffer (PBS containing 1% FBS, 2 mM EDTA and 0.05% NaN3), harvested in FACS buffer containing 0.5 μg / mL propidium iodide (Sigma, Cat. P4864), and processed on a flow cytometer (CANTO) equipped with a high-throughput sampler. tm Analysis was performed in II, BD Biosciences. Live HEK#354 cells were gated into a propidium iodide-negative population and Fpn-GFP expression was analyzed. The MFI of Fpn-GFP for >2000 live cells diluted with each compound was calculated using FlowJo (Tree Star's, Oregon), and the potency of Fpn inhibitors in inducing Fpn-GFP internalization and degradation was calculated according to the description of the hepcidin internalization assay. The EC50 of compounds (II) and (II') tested by FACS in the assay of membrane iron transporter internalization and degradation was...50 The data are listed in Table 4. In this determination, the average EC of ferricyanide was... 50 The value is 0.004μM±0.002μM.

[0902] Table 4 shows the average (AVE) EC of compounds (II) and (II') tested in the membrane iron transporter internalization and degradation assay. 50 The data is used for multiple measurements.

[0903] sequence list <110> Weaver (International) Co., Ltd. <120> Preparation method of membrane iron transporter inhibitor <130> H67465WO <150> EP20165358.1 <151> 2020-03-24 <160> 2 <170> BiSSAP 1.3.6 <210> 1 <211> twenty one <212> DNA <213> people <220> <223> forward primer <400> 1 caggtttgtg agcatcctga a 21 <210> 2 <211> 19 <212> DNA <213> people <220> <223> reverse primer <400> 2 ggcggcgact aaggagagg 19

Claims

1. A method for preparing compounds of general formula (I) and pharmaceutically acceptable salts thereof, include The compound of formula (IM-3) reacts with the compound of formula (RM-3) under alkaline conditions. To provide compounds of formula (I); in X 1 It is N, and X 2 It is O or S, forming a functional group. ； or X 1 Is it O or S, and X? 2 It is N, forming a group. or , In each case, * indicates the bonding position with the carbonyl group, and ** indicates the second bonding position; and m is an integer 1; n is an integer of 1 or 2; o is the integer 1; A represents the CH group; R 1 and R 2 Each represents hydrogen; R 3 It represents hydrogen; R 4 It represents hydrogen; R 5 The meaning of having the following groups: in * indicates the bonding location, and R y Selected from C 1-3 Alkyl, halogen and C 1-3 Halogenated alkyl groups; and R 6 It represents hydrogen.

2. The method for preparing compounds of general formula (I) and pharmaceutically acceptable salts thereof according to claim 1, further comprising the following steps: The compound of formula (IM-2) is reacted with the compound of formula (RM-2) to form the compound of formula (IM-3). in X 1 X 2 o, A, R 1 R 4 and R 5 It has the meaning as defined in claim 1.

3. The method according to claim 2, further comprising the following steps: Compound (IM-2) was prepared by converting compound (RM-1) to compound (IM-1) followed by ester cleavage: in R y’ express -hydrogen, or - halogens; and X 1 X 2 A and R 4 It has the meaning as defined in claims 1 and 2.

4. The method according to claim 3, wherein The compound of formula (IM-1) is prepared according to one of the following reaction schemes: Reaction scheme a): ;or Reaction scheme b): ;or Reaction scheme c): ； Where X 1 X 2 A and R 4 It has the meaning as defined in any one of claims 1 to 3.

5. The method according to claim 3 or 4, wherein The preparation of compound (IM-2) from compound (RM-1) via intermediate compound (IM-1) is carried out in a one-pot reaction in a single process step.

6. The method according to claim 1 or 2, wherein In group R 5 In this context, the Ry substituents are selected from fluorine and bromine.

7. The method according to claim 1 or 2, comprising the following reaction steps: Step 1: Step 2: Step 3: Where X 1 X 2 R 3 R 4 R 6 R y A, m, n and o have the meanings as defined in any of the preceding claims.

8. The method according to claim 1 or 2, wherein the compound of formula (II) and its pharmaceutically acceptable salt are prepared. (II) The following steps are included: Step 1-a: Step 2-a: Step 3-a: 。 9. The method according to claim 1 or 2, wherein the compound of formula (II') and its pharmaceutically acceptable salt are prepared. (II') The following steps are included: Step 1-a: Step 2-a: Step 3-a: 。 10. The method according to claim 1 or 2, further comprising the step of: Compounds of formula (I), (II), or (II') can be converted into their pharmaceutically acceptable salts using appropriate bases or acids and / or solvents.

11. The method according to claim 10, wherein in the step of converting the compound of formula (I) or (II) or (II') into a pharmaceutically acceptable salt thereof, an acid selected from the group consisting of benzoic acid, citric acid, fumaric acid, hydrochloric acid, lactic acid, malic acid, maleic acid, methanesulfonic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, and toluenesulfonic acid is used.

12. The method according to claim 10, wherein the compound of formula (I) or (II) or (II') is converted into a salt, said salt being selected from monoHCl salts (1HCl), triHCl salts (3HCl), H2SO4 salts, 0.5 H3PO4 salts and 1H3PO4 salts.

13. The method of claim 7, wherein steps 1 and 2 are each carried out in a condensed one-pot reaction, and / or, if present, the step of converting intermediate compound IM-3 into a pharmaceutically acceptable salt of compound (I) is carried out in a condensed one-pot reaction.

14. The method of claim 8, wherein steps 1-a and 2-a are each carried out in a condensed one-pot reaction, and / or, if present, the step of converting intermediate compound IM-3-b into a pharmaceutically acceptable salt of compound (II) is carried out in a condensed one-pot reaction.

15. Intermediate compounds of general formula (IM-3) in X 1 For N and X 2 It is O or S, forming a group or X 1 For O or S and X 2 For N, form a group or , In each case, * indicates the bonding position with the carbonyl group, and ** indicates the second bonding position; and R 1 R 4 R 5 A and o have the meanings as defined in any of the preceding claims.

16. The intermediate compound according to claim 15, represented by the general formula (IM-3-b). 。 17. The intermediate compound according to claim 15, represented by the general formula (IM-3-b'). 。 18. A method for preparing the intermediate compound (IM-3), (IM-3-b) or (IM-3-b') according to claims 15-17, comprising the reaction steps defined in any one of claims 2 to 9.

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