Substituted bicyclic systems as HSET inhibitors

Abstract

The present invention relates to substituted bicyclic systems of general formula I, and to the use of the compounds of the invention for the treatment and / or prevention of hyperproliferative diseases and disorders in mammals, particularly humans, and to pharmaceutical compositions containing such compounds.

Description

[Technical Field]

[0001] The present invention relates to compounds of general formula I, [ka] and uses of the compounds of this invention for the treatment and / or prevention of hyperproliferative diseases and disorders (such as cancer) in mammals, particularly humans, and pharmaceutical compositions containing such compounds. [Background technology]

[0002] Background of the Invention DNA replication, followed by equal chromosome segregation, ensures the accurate transmission of genetic information to daughter cells (Hall et al., 2003; Nigg, 2002; Zyss and Gergely, 2009). In most normal and malignant cells, the centrosome acts as the dominant site for spindle pole formation (Meunier and Vernos, 2012). Centrosome duplication is also tightly regulated and occurs concomitantly with DNA replication, ensuring the generation of two functional centrosomes that form the poles of the mitotic spindle (Sharp et al., 2000). Microtubule (MT) motor proteins play a central role in the assembly of a functional mitotic spindle (Cai et al., 2010; Ganem and Compton, 2004). One such protein, the minus-end MT motor HSET (encoded by KIFC1 in humans and Kifc5a in mice), has attracted attention in cancer due to its effects on cell division (Cai et al., 2010; Goshima et al., 2005).

[0003] Although the importance of centrosomes, particularly HSET, for bipolar spindle formation has received considerable attention in recent years, the precise role of HSET in this process remains a topic of debate (Mahoney et al., 2006; Tillement et al., 2009). Recent reports have linked centrosome amplification and high HSET expression to chromosome missegregation and aneuploidy, hallmarks of human cancer (Marx et al., 2009). Centrosome amplification disrupts asymmetric cell division in neuroblastoma cells and drives tumorigenesis in fly models (Basto et al., 2008). Supernumerary centrosomes are also found in most solid tumor types and constitute a marker of malignancy in breast, brain, prostate, cervical, kidney, and bladder cancers (Chan, 2011). Thus, it is becoming increasingly clear that supernumerary centrosomes are not only a sign of malignancy but can also lead to neoplastic transformation (Ogden et al., 2013). However, not all cells with amplified centrosomes undergo multipolar mitosis, and centrosome clustering is a key mechanism by which cells with extra centrosomes acquire a pseudo-bipolar spindle (Basto et al., 2008; Ganem et al., 2009).

[0004] Although centrosome clustering prevents multipolar division and cell death, it prolongs mitosis and increases the frequency of chromosome missegregation as a result of merotelic kinetochore attachment (Ganem et al., 2009; Kwon et al., 2008; Yang et al., 2008). Based on previous studies, centrosome clustering may prove a weakness in cancer cells with supernumerary centrosomes (Basto et al., 2008), and growing evidence suggests that inhibiting centrosome clustering may offer a novel therapeutic strategy for tumors with a high incidence of centrosome amplification (Jordan and Wilson, 2004; Ogden et al., 2012).

[0005] A key protein known to be essential for centrosome clustering is HSET (Ncd in flies). HSET is required for tumor cell clustering of supernumerary centrosomes (Basto et al., 2008; Kwon et al., 2008). HSET is a member of the kinesin-14 family of MT motor proteins, a force-generating enzyme that facilitates movement along intracellular MTs (Mountain et al., 1999) and transports organelles, protein complexes, and mRNAs along microtubules in an ATP-dependent manner. HSET is a minus-end-directed kinesin motor that exerts inward force by cross-linking and sliding microtubules (Walczak et al., 1997; Cai et al., 2009; Rath et al., 2012). Although the exact role of HSET in cell division remains unclear, previous evidence suggests that HSET is essential for the survival of cancer cells but not for the survival of normal cells (Ganem et al., 2009; Kwon et al., 2008). High HSET expression levels are strongly correlated with brain metastasis of non-small cell lung cancer, suggesting a link between HSET, centrosome amplification, and tumorigenesis (Cai et al., 2010; Gordon et al., 2001; Grinberg-Rashi et al., 2009). Knockdown of HSET in normal retinal pigment epithelial 1 (RPE-1) cells or the breast cancer cell line MCF-7 (which do not have a high incidence of centrosome amplification) does not inhibit bipolar spindle formation, and the cells undergo normal division (Kleylein-Sohn et al., 2012; Kwon et al., 2008). In contrast, knockdown of HSET in breast cancer and neuroblastoma cell lines MDA-MB-231 and N1E-115, which contain supernumerary centrosomes, respectively, prevents centrosome clustering and induces cell death by multipolar anaphases (Kwon et al., 2008).Therefore, the above findings suggest HSET as a target of interest in cancer treatment (Basto et al., 2008; Kraljevic Pavelic et al., 2011; Kramer et al., 2011; Kwon et al., 2008).

[0006] Multiple studies have shown that HSET deficiency increases the frequency of cell death and multipolarization in cells with supernumerary centrosomes, but not in normal cells. For example, HSET deficiency induces spindle multipolarity and selectively sensitizes centrosome-amplified ER- breast cancer cell lines, including triple-negative breast cancer (TNBC), to cell death (Patel et al., 2018). HSET deficiency has been identified as a selective inducer of cytotoxicity in centrosome-amplified cancer cells (Drosopoulos et al., 2014). Additionally, HSET overexpression correlates with poor prognosis and resistance to docetaxel in breast cancer (De et al., 2009; Li et al., 2015), ovarian adenocarcinoma patients (Pawar et al., 2014), and numerous other cancer types (Pannu et al., 2015). Furthermore, in non-small cell lung cancer (NSCLC), HSET expression was found to be highly predictive of the presence of brain metastases in both early and advanced disease (Grinberg-Rashi et al., 2009).

[0007] A wide range of tumors, including those with centrosome amplification, are treated with cytotoxic microtubule-targeting drugs (e.g., taxol, eribulin). Although these drugs induce temporary remissions, they typically exhibit severe side effects and the development of drug resistance, leading to early relapse. More recently, agents targeting kinesin motor proteins, such as Eg5 inhibitors, have been investigated to treat various human tumors (which induce monopolar spindles, a phenotype opposite to HSET inhibition) and to target all rapidly dividing cells (including myeloid cells). Consequently, they suffer from dose-limiting toxicity, similar to other antimitotic treatments. In contrast, HSET inhibitors are expected to exhibit reduced toxicity by selectively killing cells with centrosome amplification, while cells with a normal number of centrosomes would remain unaffected (Ganem et al., 2009; Patel et al., 2015). Together, these data provide support for developing drugs that selectively inhibit HSET to target centrosome-amplified tumors ( Myers and Collins, 2016 ).

[0008] Examples of small molecule HSET inhibitors have been described in the literature. AZ82 is an ADP / ATP-competitive inhibitor that has been shown to be selective for a panel of nine other kinesins, including Eg5 (Wu et al., 2013). AZ82 inhibits microtubule-stimulated HSET ATPase activity in biochemical assays (IC 50 = 0.3 μM), induced the formation of multipolar spindles and mitotic catastrophe in cells with amplified centrosomes. CW069 was an inhibitor of HSET (IC 50 = 75 μM) (Watts et al., 2013). SR31527 also inhibited HSET biochemically (IC 50 = 6.6 μM) (Zhang et al., 2016). More information can be found in WO09155025 and WO15085088.

[0009] Thus, there remains a need for therapies for the treatment and prevention of hyperproliferative diseases and disorders such as cancer. Therefore, the goal was to find HSET inhibitors that are potential therapeutic agents for the treatment of cancer diseases. Summary of the Invention

[0010] SUMMARY OF THE INVENTION It has surprisingly been found that the compounds according to the present invention are highly selective and effective inhibitors of HSET and thus the compounds of the present invention can be used in the treatment of hyperproliferative diseases and disorders such as cancer.

[0011] The present invention relates to compounds of general formula I, [ka] During the ceremony W is [ka] [ka] represents R 1 is NO2, COOA, A, OA, NHA, NHCOA, CONHA, CONA2, COA or R 4 represents R 2 teeth, [ka] Represents and represents, R 3 represents H or A, R 4 represents H, oxadiazolyl, tetrazolyl, pyrazolyl, oxazolyl or isoxazolyl which is unsubstituted or substituted by unbranched or branched alkyl having 1 to 4 C atoms, A represents an unbranched or branched alkyl or cycloalkyl having 1 to 10 C atoms, wherein two adjacent CH and / or CH groups may form a double bond, and wherein one or two non-adjacent CH and / or CH groups may be replaced by an N atom, an O atom, and / or an S atom, and wherein 1 to 7 H atoms may be replaced by an F or CI atom; Hal denotes F, Cl, Br, or I; and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any proportion.

[0012] A preferred embodiment of the present invention is a compound of formula I, wherein W is [ka] represents and R 1 , R 2 , R 3 , R 4 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in all proportions.

[0013] A preferred embodiment of the present invention is a compound of formula I, wherein W is [ka] represents and R 1 , R 2 , R 3 , R 4 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in all proportions.

[0014] A preferred embodiment of the present invention is a compound of formula I, wherein R 1 COOA, OA or R 4 represents As well as W, R 2 , R 3 , R 4 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in all proportions.

[0015] A preferred embodiment of the present invention is a compound of formula I, wherein R 1 represents COOA, As well as W, R 2 , R 3 , R 4 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in all proportions.

[0016] A preferred embodiment of the present invention is a compound of formula I, wherein R 2 teeth, [ka] represents As well as W, R 1 , R 3 , R 4 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in all proportions.

[0017] A preferred embodiment of the present invention is a compound of formula I, wherein R 2 teeth, [ka] represents As well as W, R1 , R 3 , R 4 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in all proportions.

[0018] A preferred embodiment of the present invention is a compound of formula I, wherein R 4 represents oxadiazolyl, oxazolyl, or tetrazolyl, and W, R 1 , R 2 , R 3 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in all proportions.

[0019] A preferred embodiment of the present invention is a compound of formula I, wherein R 4 represents oxadiazolyl, and W, R 1 , R 2 , R 3 and A are compounds having the meanings as disclosed above, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in all proportions.

[0020] The present invention preferably relates to a compound selected from the group consisting of: [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any proportion.

[0021] Furthermore, the following abbreviations have the following meanings: Boc ter-butoxycarbonyl CBZ benzyloxycarbonyl DNP 2,4-dinitrophenyl FMOC 9-Fluorenylmethoxycarbonyl imi-DNP 2,4-dinitrophenyl at the 1-position of the imidazole ring Ome methyl ester POA Phenoxyacetyl DCCI Dicyclohexylcarbodiimide HOBt 1-hydroxybenzotriazole

[0022] The present invention further relates to pharmaceutical preparations comprising one or more compounds according to the invention and / or one of their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, as well as mixtures thereof in any ratio.

[0023] The invention also relates to pharmaceutical preparations according to the invention of this type which contain further excipients and / or adjuvants.

[0024] Additionally, the present invention also relates to the above pharmaceutical preparations according to the invention, which comprise at least one further medicament active compound.

[0025] Pharmaceutically or physiologically acceptable derivatives are understood to mean, for example, salts of the compounds of the present invention, as well as so-called prodrug compounds. Prodrug compounds are understood to mean derivatives of the compounds of the present invention that are modified, for example, with alkyl or acyl groups (see also amino- and hydroxyl-protecting groups below), sugars, or oligopeptides, and that are rapidly cleaved or released in the organism to form active molecules. These also include biodegradable polymer derivatives of the compounds of the present invention, as described, for example, in Int. J. Pharm. 115 (1995), 61-67.

[0026] The compounds of the present invention can be used in their final non-salt form. On the other hand, the present invention also covers the use of the compounds of the present invention in the form of their pharmaceutically acceptable salts, which can be derived from various organic and inorganic bases by procedures known in the art. Most of the pharmaceutically acceptable salt forms of the compounds of the present invention are prepared by conventional methods. When the compounds of the present invention contain a carboxyl group, a suitable salt can be formed by reacting the compounds of the present invention with a suitable base to give the corresponding base addition salt. Such bases include, for example, alkali metal hydroxides, including potassium hydroxide, sodium hydroxide, and lithium hydroxide; alkaline earth metal hydroxides, such as barium hydroxide and calcium hydroxide; alkali metal alkoxides, such as potassium ethoxide and sodium propoxide; and various organic bases, such as piperidine, diethanolamine, and N-methylglutamine. Aluminum salts of the compounds of the present invention are also included.

[0027] Furthermore, base salts of the compounds of the present invention include, but are not intended to represent a limitation, aluminum, ammonium, calcium, copper, iron(III), iron(II), lithium, magnesium, manganese(III), manganese(II), potassium, sodium, and zinc salts.

[0028] Of the above salts, preferred are ammonium; the alkali metal salts sodium and potassium, and the alkaline earth metal salts calcium and magnesium. Salts of the compounds of the present invention derived from pharmaceutically acceptable organic non-toxic bases include primary amines, secondary amines, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, chloroprocaine, choline, N,N'-dibenzylethylenediamine (benzathine), dicyclohexylamine, diethanolamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethyl ... These include, but are not intended to represent a limitation, salts of diamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lidocaine, lysine, meglumine, N-methyl-D-glucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethanolamine, triethylamine, trimethylamine, tripropylamine, and tris(hydroxymethyl)methylamine (tromethamine).

[0029] As mentioned, pharmaceutically acceptable base addition salts of the compounds of the present invention are formed with metals or amines, such as alkali metals and alkaline earth metals or organic amines. Preferred metals are sodium, potassium, magnesium, and calcium. Preferred organic amines are N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methyl-D-glucamine, and procaine.

[0030] Base addition salts of compounds of the invention are prepared by bringing the free acid form into contact with a sufficient amount of the desired base, causing the formation of the salt in a conventional manner. The free acid may be regenerated by bringing the salt form into contact with an acid and isolating the free acid in a conventional manner. The free acid forms differ in some respects from their corresponding salt forms with respect to certain physical properties, such as solubility in polar solvents; however, for purposes of this invention, the salts otherwise correspond to their respective free acid forms.

[0031] In view of the above definition, it can be seen that in this context the term "pharmaceutically acceptable salt" is understood to mean the active compound of the present invention, including the compound of the present invention in one of its salt forms (especially when this salt form confers improved pharmacokinetic properties to the active compound compared to the free form of the active compound or any other salt form of the active compound previously used).The pharmaceutically acceptable salt form of the active compound can also provide the active compound with desired pharmacokinetic properties that it did not previously have, and can even have a positive effect on the pharmacodynamics of the active compound in relation to its therapeutic effectiveness in the body.

[0032] Solvates of the compounds of the invention are understood to mean adducts of the compounds of the invention with inert solvent molecules, which form owing to their mutual attractive force. Solvates are, for example, hydrates, such as mono- or dihydrates, or alcoholates, i.e., addition compounds with alcohols, such as, for example, methanol or ethanol.

[0033] All physiologically acceptable salts, derivatives, solvates and stereoisomers of these compounds, as well as mixtures thereof in any proportion, also come into accordance with the invention.

[0034] Because the compounds of the present invention may contain one or more centers of chirality, all stereoisomers, enantiomers, diastereomers, etc. of the compounds of the present invention are also claimed in the present invention.

[0035] The invention also relates to the optically active forms (stereoisomers), the enantiomers, the racemates, the diastereomers and the hydrates and solvates of these compounds.

[0036] The compounds of the present invention may be chiral due to their molecular structure and may consequently exist in various enantiomeric forms. They may therefore be in racemic or optically active form. Since the pharmaceutical efficacy of the racemates or stereoisomers of the compounds of the present invention may differ, it may be desirable to use enantiomers. In these cases, the final products, and even intermediates, may be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art or already employed as such in synthesis.

[0037] Pharmaceutically or physiologically acceptable derivatives are understood to mean, for example, salts of the compounds according to the invention, as well as so-called prodrug compounds. Prodrug compounds are understood to mean compounds of the invention that are modified, for example, with alkyl or acyl groups (see also amino- and hydroxyl-protecting groups below), sugars, or oligopeptides, and are rapidly cleaved or liberated in the organism to form effective compounds according to the invention. These also include biodegradable polymer derivatives of the compounds according to the invention, as described, for example, in Int. J. Pharm. 115 (1995), 61-67.

[0038] Suitable acid addition salts are all physiologically or pharmacologically acceptable inorganic or organic salts of acids, such as halides, especially hydrochlorides or hydrobromides, lactates, sulfates, citrates, tartrates, maleates, fumarates, oxalates, acetates, phosphates, methylsulfonates or p-toluenesulfonates.

[0039] Very particular preference is given to the hydrochlorides, trifluoroacetates or bistrifluoroacetates of the compounds according to the invention.

[0040] Solvates of the compounds of the invention are understood to mean adducts of inert solvent molecules onto the compounds of the invention, which form as a result of their mutual attractive force. Solvates are, for example, hydrates, such as mono- or dihydrates, or alcoholates, i.e., addition compounds with alcohols (such as, for example, methanol or ethanol).

[0041] Furthermore, the compounds of the present invention are intended to include isotopically labeled forms thereof. Isotopically labeled forms of the compounds of the present invention are identical to the compounds except for the fact that one or more atoms of the compound have been replaced by an atom or atoms having an atomic mass or mass number different from the atomic mass or mass number of most naturally occurring atoms. Examples of isotopes that are readily commercially available and can be incorporated into the compounds of the present invention by well-known methods include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example: 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F, and 36 CI, respectively. Compounds of the present invention, prodrugs thereof, or pharmaceutically acceptable salts of either thereof, that contain one or more of the above isotopes and / or other isotopes of other atoms are intended to be part of the present invention. Isotopically labeled compounds of the present invention can be used in a number of beneficial ways. For example, 3 H or 14Isotopically labeled compounds of the present invention incorporating radioactive isotopes such as C are suitable for use in medicament and / or substrate tissue distribution assays. 3 H) and carbon-14 ( 14 C) are particularly preferred due to their simple preparation and excellent detectability. Heavier isotopes (e.g., deuterium ( 2 Incorporation of H) into compounds of the present invention has therapeutic advantages due to the higher metabolic stability of the isotopically labeled compounds. Higher metabolic stability translates directly into increased in vivo half-life or lower dosages, which will represent a preferred embodiment of the present invention under most circumstances. Isotopically labeled compounds of the present invention can generally be prepared by carrying out the procedures disclosed in the synthetic schemes and related descriptions herein, in the Examples section, and in the Preparations section, substituting readily available isotopically labeled reactants for non-isotopically labeled reactants.

[0042] Deuterium ( 2 H) can also be incorporated into the compounds of the present invention. The primary kinetic isotope effect is the change in the rate of a chemical reaction due to the exchange of an isotope nucleus, which is caused secondarily by the change in the ground state energy required for covalent bond formation after this isotope exchange. The exchange of a heavier isotope usually results in a lowering of the ground state energy for the chemical bond, which in turn causes a reduction in the rate of rate-limiting bond breaking. If the bond breaking occurs in or near a saddle point region along the coordinate of a multi-product reaction, the product distribution ratio can be substantially altered. To illustrate: when deuterium is attached to a carbon atom at a non-exchangeable position, k M / k D A rate difference of 2 to 7 is typical. If this rate difference is successfully applied to an oxidation-prone compound of the present invention, the in vivo profile of the compound may thereby be dramatically altered, resulting in improved pharmacokinetic properties.

[0043] When discovering and developing therapeutic agents, those skilled in the art attempt to optimize pharmacokinetic parameters while retaining desired in vitro properties. It is reasonable to assume that many compounds with poor pharmacokinetic profiles are prone to oxidative metabolism. Currently available in vitro liver microsomal assays provide valuable information about the course of this type of oxidative metabolism, which in turn allows for the rational design of deuterated compounds of the present invention with improved stability through resistance to such oxidative metabolism. Significant improvements in the pharmacokinetic profiles of the compounds of the present invention are thereby obtained, improving in vivo half-life (T1 / 2), the concentration at maximum therapeutic effect (C max ), area under the dose-response curve (AUC), and F; and quantitatively in terms of reduced clearance, dose, and material cost.

[0044] The following is intended to illustrate what has been described above: Compounds of the present invention having multiple potential attack sites for oxidative metabolism (e.g., benzylic hydrogen atoms and hydrogen atoms attached to nitrogen atoms) are prepared as a series of analogs in which various combinations of hydrogen atoms are replaced with deuterium atoms (so that some, most, or all of these hydrogen atoms can be replaced with deuterium atoms). Determination of half-life allows for a convenient and accurate determination of the extent to which resistance to oxidative metabolism has been improved. In this way, it is determined that the half-life of the parent compound can be extended by up to 100% as a result of this type of deuterium-hydrogen exchange.

[0045] The replacement of hydrogen with deuterium in the compounds of the invention can also be used to achieve favorable modification of the metabolic spectrum of the starting compound to reduce or eliminate undesired toxic metabolites. For example, if a toxic metabolite arises through oxidative carbon-hydrogen (CH) bond cleavage, it can be reasonably expected that a deuterated analog will significantly reduce or eliminate the production of the undesired metabolite, even if the specific oxidation is not the rate-limiting step. Further state-of-the-art information on deuterium-hydrogen exchange is given, for example, in Hanzlik et al., J. Org. Chem. 55, 3992-3997, 1990; Reider et al., J. Org. Chem. 52, 3326-3334, 1987; Foster, Adv. Drug Res. 14, 1-40, 1985; Gillette et al., Biochemistry 33(10), 2927-2937, 1994; and Jarman et al. Carcinogenesis 16(4), 683-688, 1993.

[0046] The present invention also relates to mixtures of the compounds of the present invention according to the present invention, for example mixtures of two diastereomers (for example in a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:10, 1:100, 1:100 or 1:1000). These are particularly preferably mixtures of two stereoisomeric compounds. However, preference is also given to mixtures of two or more compounds of the present invention.

[0047] Additionally, the present invention relates to a process for the preparation of the compounds of the invention, characterized in that a) a base of a compound of the invention is converted into one of its salts by treatment with an acid, or b) An acid of a compound of the invention is converted into one of its salts by treatment with a base.

[0048] It is also possible in each case to carry out the reactions stepwise and to adapt the protecting group concept to modify the order of ligation of the building blocks.

[0049] The starting materials or compounds are generally known. If they are novel, they can be prepared by methods known per se.

[0050] If desired, the starting materials can also be formed in situ not by isolating them from the reaction mixture, but instead by immediately converting them further to the compounds of the invention.

[0051] The compounds of the present invention are preferably obtained by liberating them from their functional derivatives by solvolysis, especially by hydrolysis or hydrogenolysis.Preferred starting materials for solvolysis or hydrogenolysis contain correspondingly protected amino, carboxyl, and / or hydroxyl groups in place of one or more free amino, carboxyl, and / or hydroxyl groups, preferably those bearing an amino-protecting group in place of the H atom connected to the N atom.Furthermore, preferred starting materials are those bearing a hydroxyl-protecting group in place of the H atom of the hydroxyl group.Preferred are also starting materials bearing a protected carboxyl group in place of a free carboxyl group.It is also possible for multiple identically or differently protected amino, carboxyl, and / or hydroxyl groups to be present in the starting material molecule.When the protecting groups present are different from each other, they can in many cases be selectively cleaved off.

[0052] The term "amino-protecting group" is generally known and refers to a group suitable for protecting (blocking) an amino group against chemical reaction, but which can be easily removed after the desired chemical reaction has been carried out elsewhere in the molecule. Typical of such groups are, inter alia, unsubstituted or substituted acyl groups, as well as unsubstituted or substituted aryl groups (e.g., 2,4-dinitrophenyl) or aralkyl groups (e.g., benzyl, 4-nitrobenzyl, triphenylmethyl). While the amino-protecting group is removed after the desired reaction or reaction sequence, its type and size are not critical, but preferred are those having 1-20, especially 1-8, carbon atoms. The term "acyl group" should be understood in the broadest sense in relation to the present process. It encompasses acyl groups derived from aliphatic, araliphatic, aromatic, or heterocyclic carboxylic or sulfonic acids, inter alia, alkoxycarbonyl groups, aryloxycarbonyl groups, and especially aralkoxycarbonyl groups. Examples of such acyl groups are alkanoyl, such as acetyl, propionyl, buturyl, aralkanoyl, such as phenylacetyl, aroyl, such as benzoyl or toluyl, aryloxyalkanoyl, such as phenoxyacetyl, alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, BOC, 2-iodoethoxycarbonyl, aralkoxycarbonyl, such as CBZ, 4-methoxybenzyloxycarbonyl, or FMOC. Preferred acyl groups are CBZ, FMOC, benzyl, and acetyl.

[0053] The terms "acid protecting group" or "carboxy protecting group" are also commonly known and refer to groups that are suitable for protecting (blocking) the -COOH group against chemical reaction, but which can be easily removed after the desired chemical reaction has been carried out elsewhere in the molecule.

[0054] Instead of the free acid, it is typical to use an ester, such as substituted and unsubstituted alkyl esters (such as methyl, ethyl, tert-butyl, and their substituted derivatives), substituted and unsubstituted benzyl esters or silyl esters. The type and size of the acid protecting group are not critical, but preferred are those having 1-20, especially 1-10, C atoms.

[0055] The term "hydroxyl protecting group" is also commonly known and refers to a group suitable for protecting (blocking) a hydroxyl group against chemical reaction, but which can be easily removed after the desired chemical reaction has been carried out elsewhere in the molecule. Typical of such groups are the above-mentioned unsubstituted or substituted aryl, aralkyl, or acyl groups, as well as alkyl groups. The type and size of the hydroxyl protecting groups are not critical, but preferred are those having 1 to 20, especially 1 to 10, carbon atoms. Examples of hydroxyl protecting groups are, among others, benzyl, p-nitrobenzoyl, p-toluenesulfonyl, and acetyl, where benzyl and acetyl are preferred.

[0056] Further representative examples of amino-, acid-, and hydroxyl-protecting groups can be found, for example, in "Greene's Protective Groups in Organic Synthesis", fourth edition, Wiley-Interscience, 2007.

[0057] Functional derivatives of the compounds of the invention, which are to be used as starting materials, may be prepared by known methods of amino acid and peptide synthesis, for example as described in the standard works and patent applications.

[0058] The compounds of the present invention are liberated from their functional derivatives using, for example, strong acids, advantageously trifluoroacetic acid or perchloric acid, but also other strong inorganic acids such as hydrochloric acid or sulfuric acid, strong organic acids such as trichloroacetic acid, or sulfonic acids such as benzoyl- or p-toluenesulfonic acid, depending on the protecting group used. The presence of additional inert solvents and / or catalysts is possible, but not necessarily required.

[0059] Depending on the respective synthetic route, the starting materials may optionally be reacted in the presence of an inert solvent.

[0060] Suitable inert solvents are, for example, heptane, hexane, petroleum ether, DMSO, benzene, toluene, xylene, trichloroethylene, 1,2-dichloroethane, carbon tetrachloride, chloroform, or dichloromethane; alcohols, such as methanol, ethanol, isopropanol, n-propanol, n-butanol, or tert-butanol; ethers, such as diethyl ether, diisopropyl ether (preferably due to substitution on the indole nitrogen), tetrahydrofuran (THF), or dioxane; glycol ethers, such as ethylene glycol monomethyl or monoethyl ether, ethylene glycol dimethyl ether (diglyme); ketones, such as acetone or butanone; amides, such as acetamide, dimethylacetamide, N-methylpyrrolidone (NMP), or dimethylformamide (DMF); nitriles, such as acetonitrile; esters, such as ethyl acetate; carboxylic acids or acid anhydrides, such as acetic acid or acetic anhydride; nitro compounds, such as nitromethane or nitrobenzene, optionally in mixtures with each other or with water.

[0061] The amount of solvent is not critical; 10 g to 500 g of solvent can preferably be added per 1 g of the compound of the invention to be reacted.

[0062] It may also be advantageous to add an acid-binding agent, such as an alkali or alkaline earth metal hydroxide, carbonate, or bicarbonate, or other alkali or alkaline earth metal salt of a weak acid, preferably the potassium, sodium, or calcium salt, or an organic base, such as triethylamine, dimethylamine, pyridine, or quinoline, or an excess of an amine component.

[0063] The resulting compounds according to the present invention can be separated from the corresponding solution in which they were prepared (e.g., by centrifugation and washing) and stored in a separate composition after separation, or they can remain directly in the solution in which they were prepared. The resulting compounds according to the present invention can also be incorporated into a solvent desired for a particular application.

[0064] The reaction duration depends on the reaction conditions selected. Generally, the reaction duration is 0.5 hours to 10 days, preferably 1 to 24 hours. If microwaves are used, the reaction time can be reduced to 1 to 60 minutes.

[0065] The compounds of the present invention, as well as the starting materials for their preparation, are prepared by known methods, for example under known and suitable reaction conditions, as described in the literature (e.g., in standard works such as Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], Georg-Thieme-Verlag, Stuttgart). Variants that are known per se but are not described in more detail here can also be used here.

[0066] Conventional work-up steps, such as adding water to the reaction mixture and extracting, make it possible to obtain the compound after removal of the solvent. Further purification of the product may be advantageously followed by distillation or crystallization, or by chromatographic purification.

[0067] The acids of the present invention can be converted to the relevant addition salts using a base, for example, by reaction (including evaporation) of an equal amount of acid and base in an inert solvent such as ethanol. Suitable bases for this reaction are, in particular, those that give physiologically acceptable salts. Thus, the acids of the present invention can be converted to the corresponding metal salts, in particular alkali metal salts or alkaline earth metal salts, or the corresponding ammonium salts using a base (e.g., sodium hydroxide, potassium hydroxide, sodium carbonate, or potassium carbonate). Organic bases that give physiologically acceptable salts, such as ethanolamine, are also suitable for this reaction.

[0068] On the other hand, the bases of the present invention can be converted into the related acid addition salts using an acid, for example by reaction of an equal amount of base with an acid in an inert solvent such as ethanol (followed by evaporation). Suitable acids for this reaction are, in particular, those which give physiologically acceptable salts. Thus, inorganic acids such as sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acid such as orthophosphoric acid, sulfamic acid, and also organic acids, especially aliphatic, alicyclic, araliphatic, aromatic, or heterocyclic monobasic or polybasic carboxylic acids, sulfonic acids, or sulfuric acids, such as formic acid, acetic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methane- or ethanesulfonic acid, ethanedisulfonic acid, 2-hydroxysulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalene mono- and disulfonic acid, or lauryl sulfuric acid, can be used. Salts with physiologically unacceptable acids, such as picrates, can be used for the isolation and / or purification of the compounds of the present invention.

[0069] The compounds of the invention have been found to be well tolerated and to possess valuable pharmacological properties.

[0070] The present invention therefore also relates to the use of the compounds according to the invention for the preparation of a medicament for the treatment and / or prophylaxis of diseases which are caused, promoted and / or propagated by HSET.

[0071] The present invention therefore also relates to medicaments comprising at least one compound according to the invention and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, as well as mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions.

[0072] Especially preferred are physiological and / or pathophysiological conditions associated with HSET.

[0073] Physiological and / or pathophysiological conditions are understood to mean medically relevant physiological and / or pathophysiological conditions, such as diseases or illnesses, and medical disorders, complaints, symptoms, or complications of specific diseases, etc.

[0074] The present invention also relates to a medicament comprising at least one compound according to the invention, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions selected from the group consisting of hyperproliferative diseases and disorders.

[0075] The present invention further relates to a medicament comprising at least one compound according to the invention, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of a physiological and / or pathophysiological condition selected from the group consisting of hyperproliferative and infectious diseases and disorders, wherein the hyperproliferative disease or disorder is cancer.

[0076] The present invention therefore particularly preferably relates to medicaments comprising at least one compound according to the invention and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, as well as mixtures thereof in any proportion, wherein cancer is selected from the group consisting of acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenocortical carcinoma, bladder cancer, brain cancer, breast cancer, cervical hyperplasia, cervical cancer, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon cancer, endometrial cancer, esophageal cancer, essential thrombocytosis, genitourinary cancer, glioma, glioblastoma, hairy cell leukemia, and the like. The cancer is selected from the group consisting of hematologic malignancies, head and neck cancer, Hodgkin's disease, Kaposi's sarcoma, lung cancer, lymphoma, malignant carcinoid cancer, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, medullary thyroid carcinoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, polycythemia vera, primary brain tumor, primary macroglobulinemia, prostate cancer, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small cell lung cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, and Wilms' tumor.

[0077] The present invention further preferably relates to a medicament comprising at least one compound according to the invention and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders, wherein the hyperproliferative disease or disorder is selected from the group consisting of age-related macular degeneration, Crohn's disease, liver cirrhosis, chronic inflammation-related disorders, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retinopathy of prematurity, granulomatous diseases, immune hyperproliferation associated with organ or tissue transplantation, and immunoproliferative diseases or disorders selected from the group consisting of inflammatory bowel disease, psoriasis, rheumatoid arthritis, systemic lupus erythematosus (SLE), retinal hypoxia and vascular hyperproliferation secondary to vasculitis.

[0078] The medicaments disclosed above are intended to include the corresponding uses of the compounds according to the invention for the preparation of medicaments for the treatment and / or prophylaxis of the above physiological and / or pathophysiological conditions.

[0079] In addition, the medicaments disclosed above are intended to include corresponding methods for the treatment and / or prophylaxis of the above physiological and / or pathophysiological conditions, in which at least one compound according to the present invention is administered to a patient in need of such treatment.

[0080] The compounds according to the invention preferably exhibit advantageous biological activity that can be easily demonstrated in enzyme assays and animal experiments, as described in the examples. In such enzyme-based assays, the compounds according to the invention preferably exhibit and cause an inhibitory effect, which is usually an IC50 in the suitable range, preferably in the micromolar range, more preferably in the nanomolar range. 50 is evidenced by the values.

[0081] The compounds according to the invention can be administered to humans or animals, especially mammals such as apes, dogs, cats, rats or mice, and used in the therapeutic treatment of the human or animal body and in combating the above-mentioned diseases. They can also be used as diagnostic agents or reagents.

[0082] Furthermore, the compounds according to the present invention can be used to isolate HSET and investigate the activity or expression of HSET. Furthermore, they are particularly suitable for use in diagnostic methods for diseases associated with impaired HSET activity. The present invention therefore also relates to the use of the compounds according to the present invention for the isolation of HSET and investigation of the activity or expression of HSET, or as binders and inhibitors of HSET.

[0083] For diagnostic purposes, the compounds according to the invention may be, for example, radiolabeled. Examples of radiolabels are: 3 H, 14C. 231 I, and 125 I. A preferred labeling method is the iodogen method (Fraker et al., 1978). In addition, the compounds according to the present invention can be labeled with enzymes, fluorophores, and chemophores. Examples of enzymes are alkaline phosphatase, β-galactosidase, and glucose oxidase, examples of fluorophores are fluorescein, and examples of chemophores are luminol. For example, automated detection systems for fluorescent staining are described in US 4,125,828 and US 4,207,554.

[0084] The present invention further relates to pharmaceutical compositions containing the compounds of the present invention and their use for the treatment and / or prophylaxis of diseases and disorders in which partial or total inactivation of HSET may be beneficial.

[0085] The compounds of the present invention can be used for the preparation of pharmaceutical preparations, especially by non-chemical methods, in which they are combined with at least one solid, liquid, and / or semi-liquid excipient or adjuvant, optionally with one or more additional active compound(s), to form a suitable dosage form.

[0086] The present invention therefore also relates to pharmaceutical preparations comprising at least one compound of the invention and / or its physiologically acceptable salts, derivatives, solvates and stereoisomers, and mixtures thereof in any ratio. In particular, the present invention also relates to pharmaceutical preparations comprising further excipients and / or adjuvants, as well as to pharmaceutical preparations comprising at least one further pharmaceutically active compound.

[0087] In particular, the present invention also relates to a process for the preparation of pharmaceutical preparations, characterized in that the compound of the invention, and / or one of its physiologically acceptable salts, derivatives, solvates and stereoisomers, as well as mixtures thereof in any ratio, together with solid, liquid or semi-liquid excipients or adjuvants, and optionally together with further pharmaceutically active compounds, are brought into a suitable dosage form.

[0088] The pharmaceutical preparations according to the present invention can be used as human medicine and veterinary medicine. The patient or host can belong to any mammalian species, such as primate species, particularly humans; rodents, including mice, rats, and hamsters; rabbits; horses, cattle, dogs, cats, etc. Animal models are of interest for experimental investigations because they provide models for the treatment of human diseases.

[0089] Suitable carrier substances are organic or inorganic substances that are suitable for enteral (e.g., oral), parenteral, or topical administration and that do not react with the novel compounds, such as water, vegetable oils (such as sunflower oil or cod liver oil), benzyl alcohol, polyethylene glycol, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, lanolin, or petrolatum. Those skilled in the art, thanks to their expertise, are well aware of which adjuvants are suitable for the desired pharmaceutical preparation. In addition to solvents such as water, physiological saline solution, or alcohols such as ethanol, propanol, or glycerol, sugar solutions such as glucose or mannitol solutions, or mixtures of such solvents, gel-forming agents, tablet assistants, and other active ingredient carriers, it is also possible to use, for example, lubricants, stabilizers and / or wetting agents, emulsifiers, salts for influencing osmotic pressure, antioxidants, dispersants, antifoaming agents, buffers, flavors and / or fragrances or flavor correctors, preservatives, solubilizers, or dyes. If desired, the preparations or medicaments according to the invention may also contain one or more further active compounds, for example one or more vitamins.

[0090] If desired, the preparations or medicaments according to the invention may also comprise one or more further active compounds and / or one or more action-enhancing agents (adjuvants). The terms "pharmaceutical formulation" and "pharmaceutical preparation" are used synonymously for the purposes of this invention.

[0091] As used herein, "pharmaceutically tolerated" refers to drugs, precipitating agents, excipients, adjuvants, stabilizers, solvents, and other agents that facilitate the administration of pharmaceutical preparations therefrom to mammals without undesirable physiological side effects, such as nausea, dizziness, dyspepsia, or the like.

[0092] Pharmaceutical preparations for parenteral administration require isotonicity, euhydration, tolerability and safety (low toxicity) of the formulation, of the adjuvants used and of the primary packaging. Surprisingly, the compounds according to the present invention preferably have the advantage that they can be used directly and therefore no further purification steps for the removal of toxicologically unacceptable agents (such as high concentrations of organic solvents) or other toxicologically unacceptable adjuvants are required before using the compounds according to the present invention in pharmaceutical formulations.

[0093] The present invention also particularly preferably relates to pharmaceutical preparations comprising at least one compound according to the invention in precipitated amorphous, precipitated crystalline or dissolved or suspended form, and optionally excipients and / or adjuvants and / or further pharmaceutical active compounds.

[0094] The compounds according to the invention preferably allow for the preparation of highly concentrated formulations without causing unwanted or undesirable aggregation of the compounds according to the invention. Thus, ready-to-use solutions with high active ingredient contents can be prepared by utilizing the compounds according to the invention with or in aqueous media.

[0095] The compounds, and / or their physiologically acceptable salts and solvates, may also be lyophilized and the resulting lyophilizates may be used, for example, for the preparation of injectable preparations.

[0096] Aqueous preparations can be prepared by dissolving or suspending the compound according to the present invention in an aqueous solution, and optionally adding an adjuvant.To this end, a specified volume of stock solution containing the additional adjuvant at a specified concentration is advantageously added to a solution or suspension having a specified concentration of the compound according to the present invention, and the mixture is optionally diluted with water to a pre-calculated concentration.Alternatively, the adjuvant can be added in solid form.In each case, the required amount of stock solution and / or water can be added to the resulting aqueous solution or suspension.The compound according to the present invention can also advantageously be directly dissolved or suspended in a solution containing all additional adjuvants.

[0097] Solutions or suspensions containing the compounds according to the present invention and having a pH of 4 to 10, preferably 5 to 9, and an osmolality of 250 to 350 mosmol / kg can be advantageously prepared. The pharmaceutical preparations can thus be administered directly intravenously, intraarterially, intraarticularly, subcutaneously, or transdermally, virtually painlessly. In addition, the preparations can also be added to infusion solutions, such as glucose solution, isotonic saline, or Ringer's solution, which may also contain additional active compounds, thereby allowing relatively large amounts of the active compounds to be administered.

[0098] Pharmaceutical preparations according to the invention may also contain mixtures of compounds according to the invention.

[0099] Preparations according to the invention are physiologically well tolerated, easy to prepare, can be accurately dispensed, and are preferably stable with respect to assay, decomposition products, and aggregates throughout storage and transport and during multiple freeze-thaw processes. They can preferably be stored in a stable manner at refrigerator temperature (2-8°C), rt (23-27°C), and 60% relative atmospheric humidity (RH) for periods of at least 3 months to 2 years.

[0100] For example, the compounds according to the invention can be stored in a stable manner by drying and, if necessary, converted into ready-to-use pharmaceutical preparations by dissolution or suspension. Possible drying methods are, for example and without being limited to these examples, nitrogen gas drying, vacuum oven drying, freeze drying, washing with organic solvents followed by air drying, liquid bed drying, fluidized bed drying, spray drying, roller drying, layer drying, air drying at RT, and further methods.

[0101] The term "effective amount" refers to the amount of a drug or pharmaceutically active compound that elicits the biological or medical response sought or desired, for example, by a researcher or physician, in a tissue, system, animal, or human.

[0102] Additionally, the term "therapeutically effective amount" denotes an amount that has the following causal relationship: improved treatment, cure, prevention, or elimination of a disease, syndrome, condition, complaint, or disorder, or prevention of side effects, or also reduction in the progression of a disease, complaint, or disorder, compared to a corresponding subject not receiving this amount. The term "therapeutically effective amount" also encompasses amounts effective to increase normal physiological function.

[0103] When using the preparations or medicaments according to the present invention, the compounds according to the present invention and / or their physiologically acceptable salts and solvates are generally used in a dosage similar to known commercially available preparations or medicaments, preferably between 0.1 mg and 500 mg, particularly between 5 mg and 300 mg per unit of use. The daily dose is preferably between 0.001 mg / kg and 250 mg / kg of body weight, particularly between 0.01 mg / kg and 100 mg / kg of body weight. The preparations can be administered one or more times per day, for example, two, three, or four times per day. However, the individual dose for each patient depends on many individual factors, such as the effectiveness of the specific compound used, age, body weight, general health, sex, nutrition, the timing and method of administration, excretion rate, combination with other medications, and the severity and duration of the specific disease.

[0104] The measure of the uptake of a pharmaceutically active compound in an organism is its bioavailability.When a pharmaceutically active compound is delivered to an organism intravenously in the form of an injection solution, its absolute bioavailability, i.e., the proportion of the pharmaceutical that reaches the systemic blood (i.e., the general circulation) in its unchanged form, is 100%.In the case of oral administration of a therapeutically active compound, the active compound is generally in a solid form in the formulation, and therefore must first be dissolved so that it can overcome the barrier of entry, such as the gastrointestinal tract, oral mucosa, nasal mucosa, or skin, especially the stratum corneum, or be absorbed by the body.Pharmacokinetic, i.e., bioavailability, data can be obtained similarly to the method of J. Shaffer et al., J.Pharm.Sciences, 88 (1999), 313-318. Furthermore, a medicament of this type may be prepared using one of the processes generally known in the pharmaceutical art.

[0105] The medicament may be adapted for administration via any desired suitable route, for example, oral (including buccal or sublingual), rectal, pulmonary, nasal, topical (including buccal, sublingual, or transdermal), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, and especially intraarticular) routes. Medicaments of this type may be prepared using any process known in the pharmaceutical art, for example, by combining the active compound with excipient(s) or adjuvant(s).

[0106] Parenteral administration is preferably suitable for administering the medicaments according to the invention. In the case of parenteral administration, intra-articular administration is especially preferred.

[0107] The compounds according to the invention are also suitable for the preparation of medicaments to be administered parenterally, with slow, sustained, and / or controlled release of the active compound. They are therefore also suitable for the preparation of delayed-release formulations, which are advantageous for the patient, since only relatively long time intervals between administrations are required.

[0108] Pharmaceuticals suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions containing antioxidants, buffers, bacteriostats, and solutes (which make the preparation isotonic with the blood or synovial fluid of the recipient to be treated); and aqueous and non-aqueous sterile suspensions, which may contain suspension media and thickeners. The preparations can be delivered in single-dose or multi-dose containers, such as sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) state, so that only the addition of a sterile carrier liquid (e.g., water for injection purposes) is required immediately before use. The injection solutions and suspensions prepared according to the formulations can be prepared from sterilized powders, granules, and tablets.

[0109] The compounds of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from various phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.

[0110] The compounds of the present invention can also be coupled to soluble polymers as targeted pharmaceutical excipients. Such polymers include polyvinylpyrrolidone, pyran copolymers, polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl radicals. The compounds of the present invention can also be coupled to biodegradable polymers suitable for achieving sustained drug release, such as polymers such as polylactic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates, polylactic-co-glycolic acid, dextran-methacrylate conjugates, polyphosphoesters, various polysaccharides and polyamines, as well as poly-ε-caprolactone, albumin, chitosan, collagen, or modified gelatin, and crosslinked or amphiphilic block copolymers of hydrogels.

[0111] Suitable for enteral administration (oral or rectal) are, inter alia, tablets, dragees, capsules, syrups, juices, drops or suppositories, while suitable for topical use are ointments, creams, pastes, lotions, gels, sprays, foams, aerosols, solutions (e.g., solutions in alcohols such as ethanol or isopropanol, acetonitrile, DMF, dimethylacetamide, 1,2-propanediol, or mixtures thereof with each other and / or with water) or powders. Also particularly suitable for topical use are liposomal preparations.

[0112] In the case of a formulation to give an ointment, the active compound can be employed with either a paraffin base or a water-miscible cream base. Alternatively, the active compound can be formulated into a cream with an oil-in-water cream base or a water-in-oil base.

[0113] Medicaments adapted for transdermal administration can be delivered as separate patches for widespread contact with the recipient's epidermis. Thus, for example, the active compound can be dispensed from the patch using iontophoresis, as generally described in Pharmaceutical Research, 3(6), 318 (1986).

[0114] It goes without saying that in addition to the constituents specifically mentioned above, the medicaments according to the invention may also contain other agents as are customary in the art for particular types of pharmaceutical preparations.

[0115] The present invention also relates to a set (kit) consisting of the following separate packs: a) an effective amount of a compound of the present invention, and / or its physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any ratio; and b) an effective amount of a further pharmaceutically active compound.

[0116] The set comprises suitable containers, such as a box or carton, individual bottles, bags, or ampoules, etc. The set may, for example, comprise separate ampoules each containing an effective amount of a compound of the invention, and / or its pharmaceutically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any ratio, and an effective amount of an additional pharmaceutically active compound in dissolved or lyophilized form.

[0117] Furthermore, the medicaments according to the present invention may be used to provide an additive or synergistic effect in a known treatment and / or to restore the effectiveness of an existing treatment.

[0118] In addition to the compounds according to the invention, the pharmaceutical preparations according to the invention may also contain further pharmaceutically active compounds (e.g., for use in the treatment of cancer), other antitumor drugs. For the treatment of the other diseases mentioned, the pharmaceutical preparations according to the invention may also contain, in addition to the compounds according to the invention, further pharmaceutically active compounds known to those skilled in the art for their treatment.

[0119] In one principal embodiment, a method is provided for enhancing an immune response in a host in need thereof. The immune response can be enhanced by reducing T cell tolerance, including by increasing IFN-γ release, decreasing the production or activation of regulatory T cells, or increasing the production of antigen-specific memory T cells in the host. In one embodiment, the method comprises administering a compound of the present invention to a host in combination with or alternating with an antibody. In specific subembodiments, the antibody is a therapeutic antibody. In one specific embodiment, a method is provided for enhancing the effectiveness of passive antibody therapy, comprising administering a compound of the present invention in combination with or alternating with one or more passive antibodies. This method may enhance the effectiveness of antibody therapy for the treatment of abnormal cell proliferative disorders such as cancer, or may enhance the effectiveness of therapy in the treatment or prevention of infectious diseases. The compound of the present invention may be administered in combination with or alternating with an antibody, such as rituximab, Herceptin, or Erbitux.

[0120] In another principal aspect, there is provided a method of treating or preventing abnormal cell proliferation, the method comprising administering a compound of the present invention to a host in need thereof, substantially in the absence of another anti-cancer agent.

[0121] In another main embodiment, there is provided a method of treating or preventing abnormal cell proliferation in a host in need thereof, the method comprising administering to the host a first compound of the present invention substantially in combination with a first anti-cancer agent, followed by administering a receptor antagonist that is a second compound of the present invention. In one subembodiment, the second antagonist is administered substantially in the absence of another anti-cancer agent. In another main embodiment, there is provided a method of treating or preventing abnormal cell proliferation in a host in need thereof, the method comprising administering to the host a compound of the present invention substantially in combination with a first anti-cancer agent, followed by administering a second anti-cancer agent in the absence of an antagonist.

[0122] Thus, the cancer treatments disclosed herein can be carried out as a treatment with the compounds of the present invention or in combination with surgery, radiation therapy, or chemotherapy. This type of chemotherapy can include the use of one or more active compounds from the following categories of antitumor active compounds:

[0123] (i) Antiproliferative / antineoplastic / DNA damaging active compounds and combinations thereof when used in medical oncology, such as alkylating active compounds (e.g., cisplatin, parvoplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan, and nitrosoureas); antimetabolites (e.g., antifolates, such as fluoropyrimidines, e.g., 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea, and gemcitabine); antitumor antibiotics (e.g., anthracyclines, e.g., adriamycin, benzodiazepines, benzocaine ... isin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin, and mithramycin); antimitotic active compounds (e.g., vinca alkaloids, such as vincristine, vinblastine, vindesine, and vinorelbine, and taxoids, such as taxol and taxotere); topoisomerase inhibitors (e.g., epipodophyllotoxins, such as etoposide and teniposide, amsacrine, topotecan, irinotecan, and camptothecin), and cell differentiation active compounds (e.g., all-trans retinoic acid, 13-cis-retinoic acid, and fenretinide);

[0124] (ii) cytostatically active compounds, such as antiestrogens (e.g., tamoxifen, toremifene, raloxifene, droloxifene, and iodoxifene), estrogen receptor modulators (e.g., fulvestrant), antiandrogens (e.g., bicalutamide, flutamide, nilutamide, and cyproterone acetate), LHRH antagonists or agonists (e.g., goserelin, leuprorelin, and buserelin), progesterones (e.g., megestrol acetate), aromatase inhibitors (e.g., anastrozole, letrozole, vorazole, and exemestane), and inhibitors of 5α-reductase, such as finasteride;

[0125] (iii) active compounds that inhibit cancer invasion, including metalloprotease inhibitors such as marimastat and inhibitors of urokinase-type plasminogen activator receptor function;

[0126] (iv) inhibitors of growth factor function, e.g., growth factor antibodies, growth factor receptor antibodies, e.g., the anti-erbb2 antibody trastuzumab [Herceptin™] and the anti-erbb1 antibody cetuximab [C225]), farnesyltransferase inhibitors, tyrosine kinase inhibitors, and serine / threonine kinase inhibitors, e.g., inhibitors of the epidermal growth factor family (e.g., EGFR family tyrosine kinase inhibitors), -, for example, N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, AZD1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774), and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazolin-4-amine (CI 1033)), for example, inhibitors of the platelet-derived growth factor family, and for example, inhibitors of the hepatocyte growth factor family;

[0127] (v) anti-angiogenic compounds, such as bevacizumab, angiostatin, endostatin, linomide, batimastat, captopril, cartilage derived inhibitors, genistein, interleukin 12, lavendustin, medroxyprogesterone acetate, recombinant human platelet factor 4, tecogalan, thrombospondin, TNP-470, anti-VEGF monoclonal antibodies, soluble VEGF receptor chimeric proteins, anti-VEGF receptor antibodies, anti-PDGF receptors, inhibitors of integrins, tyrosine kinase inhibitors, serine / threonine kinase inhibitors, antisense oligonucleotides, antisense oligodeoxynucleotides, siRNA, anti-VEGF aptamers, pigment epithelium-derived factors, and the like, as described in International Patent Applications WO 97 / 22596, WO 97 / 30035, WO 97 / 32856, and WO Compounds published in 98 / 13354;

[0128] (vi) vessel-destroying agents, such as combretastatin A4 and compounds published in International Patent Applications WO 99 / 02166, WO 00 / 40529, WO 00 / 41669, WO 01 / 92224, WO 02 / 04434, and WO 02 / 08213;

[0129] (vii) antisense therapy, e.g., directed against the targets mentioned above, e.g., ISIS 2503, an anti-Ras antisense;

[0130] (viii) Gene therapy approaches, such as replacement of abnormal modified genes, e.g., abnormal p53 or abnormal BRCA1 or BRCA2, GDEPT approaches (gene-directed enzyme prodrug therapy), e.g., those using cytosine deaminase, thymidine kinase, or bacterial nitroreductase enzymes, and approaches that increase a patient's resistance to chemotherapy or radiation therapy, e.g., multidrug resistance therapy; and

[0131] (ix) Immunotherapeutic approaches, including ex-vivo and in-vivo approaches to increase the immunogenicity of tumor cells in patients, such as transfection of cytokines, e.g., interleukin 2, interleukin 4, or granulocyte-macrophage colony-stimulating factor, approaches to reduce T-cell anergy, approaches using transfected immune cells, e.g., cytokine-transfected dendritic cells, approaches for the use of cytokine-transfected tumor cells, and approaches for the use of anti-idiotypic antibodies.

[0132] (x) Chemotherapeutic agents, such as abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, live BCG (BCG live), bevacizumab, bexarotene, bleomycin, bortezomib, busulfan, calucelone, camptothecin, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone, epirubicin, epoetin alfa, estramustine, etoposide, exemestane, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, and gemcitabine.

[0133] Medicaments from Table 1 may preferably, but not exclusively, be combined with compounds of the present invention. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8]

[0134] It is assumed that, even without further embodiments, a person skilled in the art would be able to use the above description to the fullest extent, and the preferred embodiments should therefore be regarded merely as a descriptive disclosure that is in any way not limiting.

[0135] The following examples are therefore intended to illustrate the invention without limiting it. Unless otherwise indicated, percentage data are expressed as percentages by weight. All temperatures are indicated in degrees Celsius. "Conventional work-up" means: adding water if necessary, adjusting the pH if necessary to a value between 2 and 10, extracting the mixture with ethyl acetate or dichloromethane depending on the composition of the final product, separating the phases, drying the organic phase over sodium sulfate or magnesium sulfate, filtering and evaporating, and purifying the product by chromatography on silica gel and / or crystallization.

[0136] Rf value on silica gel; Mass spectrometry: EI (electron impact ionization): M +, FAB (Fast Atom Bombardment): (M+H) + , THF (tetrahydrofuran), NMP (N-methylpyrrolidone), DMSO (dimethyl sulfoxide), EtOAc (ethyl acetate), MeOH (methanol), EtOH (ethanol), TLC (thin layer chromatography)

[0137] List of abbreviations AUC Area under the plasma drug concentration-time curve C max maximum plasma concentration CL Clearance CV coefficient of variation CYP cytochrome P450 DMSO dimethyl sulfoxide F Bioavailability f a Absorption rate iv intravenous LC-MS / MS Liquid Chromatography Tandem Mass Spectrometry LLOQ Lower limit of quantification NC Not calculated ND Not determined PEG polyethylene glycol Pgp permeability glycoprotein PK (pharmacokinetics) Po (oral) Rt room temperature t 1 / 2 Half-life t max The time when the maximum plasma concentration of the drug is reached UPLC Ultra High Performance Liquid Chromatography V ss Volume of distribution (at steady state) v / v volume to volume

[0138] Methods for preparing and analyzing compounds of the present invention The invention particularly relates to the following exemplary compounds, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in all proportions.

[0139] All solvents used were commercially available and were used without further purification. Reactions were typically performed using anhydrous solvents under an inert atmosphere of nitrogen or argon. Flash column chromatography was performed using silica gel 60 (0.035-0.070 mm particle size). Flash column chromatography was also performed using a Biotage purification system using a SNAP KP-Sil cartridge or in reversed-phase mode using a SNAP Ultra C18 cartridge. Microwave-assisted reactions were performed using a Biotage Initiator microwave system.

[0140] 1 H NMR spectra were recorded on a Bruker DPX-300, DRX-400, Avance II-400, Avance III HD-400, Avance II+-500, Avance III-500, Avance Neo 600, or Avance III-700 spectrometer, using the residual signal of the deuterated solvent as the internal reference.

[0141] Chemical shifts (δ) are reported in ppm relative to tetramethylsilane (TMS) referenced to the internal deuterated solvent. 1 1 H NMR data are reported as follows: chemical shifts (multiplicities, coupling constants, and hydrogen numbers).

[0142] Multiplicities are abbreviated as follows: s (singlet), d (doublet), t (triplet), q (quartet), sext (sext), hept (septet), m (multiplet), b (broad).

[0143] HPLC / MS spectra of the products were recorded on an Agilent 1100 HPLC system (1100 high pressure gradient pump, 1100 diode array detector, wavelength: 220 nm) coupled to an Agilent 1100 mass spectrometer detector (positive mode). LC-MS analysis was carried out on a SHIMADZU LC-MS machine consisting of a UFLC 20-AD system and an LCMS 2020 MS detector.

[0144] Details of the conditions applied for HPLC / MS spectra recorded on a Shimadzu LCMS-2020 system or an Agilent 1200 system: (A): Column: HALO C18, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.1%FA, Mobile phase B: ACN with 0.1%FA; Gradient: 5%B~100%B until min 1.50, hold until min 1.80, 100%B~5%B until min 1.81, stop after 2.00; Flow: 1.5mL / min.

[0145] (B): Column: HALO C18, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.05% TFA, Mobile phase B: ACN with 0.05% TFA; Gradient: 5%B~100%B until min 1.20, hold until min 1.80, 100%B~5%B until min 1.82, stop after 2.00; Flow: 1.5mL / min. (C): Column: HALO C18 90A, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.05% TFA, Mobile phase B: ACN with 0.05% TFA; Gradient: 5%B~100%B until min 1.20, hold until min 1.80, 100%B~5%B until min 1.82, stop after 2.00; Flow: 1.2mL / min.

[0146] (D): Column: Shim-pack C18, 3.0x33mm, 3.0µm; Mobile phase A: water with 5mM NH4CO3, Mobile phase B: ACN; Gradient: 10%B to 95%B until min 1.2, hold until min 1.8, 95%B to 10%B until min 1.82, stop after 2.00; Flow rate: 1.5mL / min. (E): Column: HALO C18, 3.0*30mm, 3.0μm; Mobile phase A: water with 0.05%TFA, Mobile phase B: ACN with 0.05%TFA; Gradient: 5%B~100%B until min 1.20, hold until min 1.80, 100%B~5%B until min 1.82, stop after 2.00; Flow: 1.5mL / min.

[0147] (F): Column: HALO C18 90A, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.05% TFA, Mobile phase B: ACN with 0.05% TFA; Gradient: 5%B~60%B until min 1.80, 60%B~95%B until min 2.50, hold until min 2.80, 95%B~5%B until min 2.81, stop after 3.00; Flow: 1.2mL / min. (G): Column: Chromolith HR C18 RP-18e, 4.6x50mm; Mobile phase A: water with 0.05% FA, Mobile phase B: ACN with 0.04% FA + 1% water; Gradient: 0% B to 100% B until min 2.0, hold until min 2.5, 100% B to 0% B until min 2.51, stop after 2.95; Flow: 3.3mL / min.

[0148] (H): Column: Kinetex EVO C18, 4.6x50mm, 5.0µm; Mobile phase A: water with 0.05% FA, Mobile phase B: ACN with 0.04% FA and 1% water; Gradient: 1%B to 99%B until min 0.8, 99%B to 1%B until min 1.1, stop after 1.50; Flow: 3.3mL / min. (I): Column: Kinetex EVO C18 100A, 3.0x50mm, 2.6µm; Mobile phase A: water with 0.04% NH4OH, Mobile phase B: ACN; Gradient: 10%B to 60%B in 2.2min, 60%B to 95%B in 2.7min, hold until min 3.2, 95%B to 10%B until min 3.3, stop after 3.5, Flow: 1.2mL / min.

[0149] (J): Column: HALO C18, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.05% TFA, Mobile phase B: ACN with 0.05% TFA; Gradient: 5%B~100%B until min 0.70, hold until min 1.10, 100%B~5%B until min 1.12, stop after 1.20; Flow: 1.5mL / min. (K): Column: HALO C18, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.1%FA, Mobile phase B: ACN with 0.1%FA; Gradient: 5%B~95%B until min 2.10, hold until min 2.75, 95%B~5%B until min 2.81, stop after 3.00; Flow: 1.5mL / min.

[0150] (L): Column: HALO C18, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.1%FA, Mobile phase B: ACN with 0.1%FA; Gradient: 10%B~95%B until min 1.20, hold until min 1.80, 95%B~10%B until min 1.82, stop after 2.00; Flow: 1.5mL / min. (M): Column: Sunfire C18, 3.0x100mm, 5µm; Mobile phase A: water with 0.05% FA, Mobile phase B: ACN with 0.04% FA and 1% water; Gradient: 1%B to 99%B at min 2.0, hold until min 3.5, 99%B to 1%B until min 2.71, stop after 3.5; Flow: 1.4mL / min.

[0151] (N): Column: Chromolith HR C18, 4.6x50mm, 5µm; Mobile phase A: water with 0.1% TFA, Mobile phase B: ACN with 0.1% TFA; Gradient: 1%B to 99%B until min 2.0, hold until min 2.5, 99%B to 1%B until min 2.51, stop after 2.95; Flow: 3.3mL / min. (O): Column: HALO C18, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.05% TFA, Mobile phase B: ACN with 0.05% TFA; Gradient: 5%B~50%B until min 1.80, 50%B~95%B until min 2.50, hold until min 2.80, 95%B~5%B until min 2.81, stop after 3.00; Flow: 1.5mL / min.

[0152] (P): Column: Shim-pack velox, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.1%FA, Mobile phase B: ACN with 0.1%FA; Gradient: 5%B~100%B until min 1.50, hold until min 1.80, 100%B~5%B until min 1.81, stop after 2.00; Flow: 1.5mL / min. (Q): Column: Kinetex EVO C18, 3.0 x 50 mm, 2.6 μm; Mobile phase A: 6.5 mM NH4HCO3 + NH4OH (pH 10); Mobile phase B: ACN; Gradient: 10% B to 70% B in 2.2 min, 70% B to 95% B in 2.7 min, hold until min 3.2, 95% B to 10% B until min 3.3, stop after 3.5; Flow: 1.2 mL / min.

[0153] (R): Column: HALO C18, 3.0*30mm, 2.0μm; Mobile phase A: water with 0.1%FA, Mobile phase B: ACN with 0.1%FA; Gradient: 5%B~100%B until min 1.20, hold until min 1.80, 100%B~5%B until min 1.82, stop after 2.00; Flow: 1.5mL / min. (S): Column: Kinetex EVO C18, 3.0 x 50 mm, 2.6 μm; Mobile phase A: 6.5 mM NH4HCO3 + NH4OH (pH 10); Mobile phase B: ACN; Gradient: 10% B to 95% B in 1.9 min, hold to min 2.7, 95% B to 10% B until min 2.75, stop after 3.0; Flow: 1.2 mL / min.

[0154] (T): Column: Kinetex EVO C18, 2.1x30mm, 5.0µm; Mobile phase A: 0.0375% TFA (v / v) in water; Mobile phase B: 0.01875% TFA (v / v) in acetonitrile; Gradient: 5% B to 95% B in 0.8 min, 95% B to min 1.2, stopped after 1.55 min, 5% B; Flow: 1.5mL / min. (U): Column: Kinetex EVO C18, 2.1x30mm, 5.0µm; Mobile phase A: 0.0375% TFA (v / v) in water; Mobile phase B: 0.01875% TFA (v / v) in acetonitrile; Gradient: 0%B to 60%B in 0.8min, 60%B to min 1.2, stopped after 1.55min, 0%B; Flow: 1.5mL / min.

[0155] (V): Column: Kinetex EVO C18, 2.1x30mm, 5.0µm; Mobile phase A: 0.025% NH₃·H₂O in water (v / v); Mobile phase B: acetonitrile; Gradient: 5% B to 95% B in 0.8 min, 95% B to min 1.2, 5% B to min 1.55, stopped after 1.55 min; Flow: 1.5mL / min. (W): Column: Poroshell HPH‐C18, 3.0 × 50 mm, 2.7 μm; Mobile phase A: water with 5 mM NH4CO3, Mobile phase B: ACN; Gradient: 10% B to 95% B until min 2.00, hold until min 2.70, 95% B to 10% B until min 2.75, stop after 3.00; Flow: 1.2 mL / min.

[0156] (X): Column: Poroshell HPH‐C18, 3.0 × 50 mm, 2.7 μm; Mobile phase A: water with 5 mM NH4CO3, Mobile phase B: ACN; Gradient: 10% B to 70% B until min 2.20, 70% B to 95% B until min 2.70, hold until min 3.20, 95% B to 10% B until min 3.30, stop after 3.50; Flow: 1.2 mL / min.

[0157] Details of the conditions applied to HPLC / MS spectra on an Agilent 1200 Series HPLC and diode array detector coupled to a 6210 time-of-flight mass spectrometer with dual multimode APCI / ESI sources. (AA): Analytical separation was performed on a Merck Chromolith Flash column (RP-18e, 25x2 mm) at 40°C using a flow rate of 1.5 mL / min with a 2-minute gradient elution and detection at 254 nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 5:95 (A / B) to 100:0 (A / B) over 1.25 min, 100:0 (A / B) for 0.5 min, then reversed to 5:95 (A / B) over 0.05 min, and finally 5:95 (A / B) for 0.2 min.

[0158] (AB): Analytical separation was performed on a Merck Chromolith Flash column (RP-18e, 25x2 mm) at 30°C using a flow rate of 0.75 mL / min with a 4-minute gradient elution and detection at 254 nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 5:95 (A / B) to 100:0 (A / B) over 2.5 min, 100:0 (A / B) for 1 min, then reversed to 5:95 (A / B) over 0.1 min, and finally 5:95 (A / B) for 0.4 min. Details of the conditions applied to HPLC / MS spectra on a Waters Acquity UPLC and diode array detector coupled to a Waters G2 QToF mass spectrometer with a multimode ESI / APCI source.

[0159] (AC): Analytical separation was performed on a Phenomenex Kinetex C18 column (30x2.1 mm, 2.6 u, 100 A) at 30°C using a 2-minute gradient elution at a flow rate of 0.5 mL / min and detection at 254 nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 1.25 min, 90:10 (A / B) for 0.5 min, then back to 10:90 (A / B) over 0.15 min, and finally 10:90 (A / B) for 0.1 min.

[0160] (AD): Analytical separation was performed on a Phenomenex Kinetex C18 column (30x2.1 mm, 2.6 u, 100 A) at 30°C using a 4-minute gradient elution at a flow rate of 0.3 mL / min and detection at 254 nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 3 min, 90:10 (A / B) for 0.5 min, then back to 10:90 (A / B) over 0.3 min, and finally 10:90 (A / B) for 0.2 min.

[0161] (AE): Analytical separation was performed on an Agilent Poroshell C18 column (30x2.1mm, 2.6u, 100A) at 30°C using a 2 minute gradient elution at a flow rate of 0.5mL / min and detection at 254nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 1.25 min, 90:10 (A / B) for 0.5 min, then back to 10:90 (A / B) over 0.15 min, and finally 10:90 (A / B) for 0.1 min.

[0162] (AF): Analytical separation was performed on an Agilent Poroshell C18 column (30x2.1mm, 2.6u, 100A) at 30°C using a 4-minute gradient elution at a flow rate of 0.3mL / min and detection at 254nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 3 minutes, 90:10 (A / B) for 0.5 minutes, then back to 10:90 (A / B) over 0.3 minutes, and finally 10:90 (A / B) for 0.2 minutes. Details of the conditions applied to HPLC / MS spectra on an Agilent 1260 Infinity II Series UPLC and diode array detector coupled to a 6530 quadrupole time-of-flight mass spectrometer with an Agilent Jet Stream ESI source.

[0163] (AG): Analytical separation was performed on an Agilent Poroshell C18 column (30x2.1mm, 2.6u, 100A) at 40°C using a flow rate of 0.6mL / min with a 2 minute gradient elution and detection at 254, 280, and 214nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 1.25min, 90:10 (A / B) for 0.5min, then back to 10:90 (A / B) over 0.15min, and finally 10:90 (A / B) for 0.1min.

[0164] (AH): Analytical separation was performed on an Agilent Poroshell C18 column (30x2.1mm, 2.6u, 100A) at 40°C using a flow rate of 0.4mL / min with a 4 minute gradient elution and detection at 254, 280, and 214nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 2.5 min, 90:10 (A / B) for 1 min, then back to 10:90 (A / B) over 0.3 min, and finally 10:90 (A / B) for 0.2 min.

[0165] (AI): Analytical separation was performed on a Phenomenex Kinetex C18 column (30x2.1 mm, 2.6 u, 100 A) at 40°C using a flow rate of 0.6 mL / min with a 2-minute gradient elution and detection at 254, 280, and 214 nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 1.25 min, 90:10 (A / B) for 0.5 min, then back to 10:90 (A / B) over 0.15 min, and finally 10:90 (A / B) for 0.1 min.

[0166] (AJ): Analytical separation was performed on a Phenomenex Kinetex C18 column (30x2.1 mm, 2.6 u, 100 A) at 40°C using a flow rate of 0.4 mL / min with a 4-minute gradient elution and detection at 254, 280, and 214 nm. The mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing 0.1% formic acid. The gradient elution was as follows: 10:90 (A / B) to 90:10 (A / B) over 2.5 min, 90:10 (A / B) for 1 min, then back to 10:90 (A / B) over 0.3 min, and finally 10:90 (A / B) for 0.2 min.

[0167] Synthesis of intermediates: A1: 3-(3-methyl-1,2-oxazol-5-yl)benzoic acid [ka]

[0168] A1.1: A solution of 1-(3-bromophenyl)ethan-1-one (4.75 g; 23.864 mmol) and (1,1-dimethoxyethyl)dimethylamine (9.53 g; 71.552 mmol) was stirred at 100 °C for 16 h. The resulting mixture was concentrated under vacuum. Then, NH2OH*HCl (4.98 g; 71.664 mmol) and EtOH (100.0 mL) were added, and the resulting mixture was stirred at 90 °C for 28 h. The reaction mixture was cooled to ambient temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 3:2) to give 3.6 g (63%) of 5-(3-bromophenyl)-3-methyl-1,2-oxazole as a colorless solid. HPLC / MS m / z: 238.05 [M+H] + , Rt(A): 1.18 min.

[0169] A1.2: To a stirred solution of 5-(3-bromophenyl)-3-methyl-1,2-oxazole (900.0 mg; 3.784 mmol) and EtN (1.53 g; 15.120 mmol) in DMF (10.0 mL), Pd(dppf)Cl.CHCl (309 mg; 0.378 mmol) and formyl acetate (2.63 g; 29.865 mmol) were added. The resulting mixture was stirred at 90 °C under a nitrogen atmosphere for 2 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 2:1) to yield 468.0 mg (61%) of 3-(3-methyl-1,2-oxazol-5-yl)benzoic acid as a colorless solid. HPLC / MS m / z: 204.2 [M+H] + , Rt(B): 0.73 min.

[0170] A2: 3-(5-methyl-1,3-oxazol-2-yl)benzoic acid [ka]

[0171] A2.1: To a stirred solution of 3-bromobenzoic acid (4.75 g; 23.630 mmol) and prop-2-yn-1-amine (1.30 g; 23.630 mmol) in DMF (50.0 mL), TCFH (9.95 g; 35.445 mmol) and 1-methyl-1H-imidazole (3.88 g; 47.257 mmol) were added, and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 3:1) to yield 4.51 g (80%) of 3-bromo-N-(prop-2-yn-1-yl)benzamide as a yellow solid. HPLC / MS m / z: 240.1 [M+H] + , Rt((B): 0.77 min.

[0172] A2.2: A solution of 3-bromo-N-(prop-2-yn-1-yl)benzamide (1.80 g; 7.569 mmol) and AuCl3 (114.80 mg; 0.378 mmol) in ACN (20.0 mL) was stirred at 50 °C under nitrogen atmosphere for 16 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 2:1) to give 587 g (33%) of 2-(3-bromophenyl)-5-methyl-1,3-oxazole as a colorless solid. HPLC / MS m / z: 238.0 [M+H] + , Rt(E): 1.08 min.

[0173] A2.3: 2-(3-Bromophenyl)-5-methyl-1,3-oxazole (342.70 mg; 1.439 mmol) was converted as described in A1.2. Yield: 230 mg (79%) of a colorless solid. HPLC / MS m / z: 204.15 [M+H] + , Rt(A): 0.84 min.

[0174] A3: 3-(4-methyl-1,3-oxazol-2-yl)benzoic acid [ka]

[0175] A3.1: To a stirred solution of [3-(methoxycarbonyl)phenyl]boronic acid (475.0 mg; 2.639 mmol) and 2-bromo-4-methyl-1,3-oxazole (427.5 mg; 2.639 mmol) in THF (5.0 mL) and water (0.50 mL), sodium carbonate (559.5 mg; 5.279 mmol) and Pd(dppf)Cl.CHCl (215.5 mg; 0.264 mmol) were added, and the resulting mixture was stirred at 70 °C for 16 h under a N atmosphere. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 1:1) to afford 200 mg (35%) of methyl 3-(4-methyl-1,3-oxazol-2-yl)benzoate as a yellow solid. HPLC / MS m / z: 218.2 [M+H] + , Rt(A): 1.01 min.

[0176] A3.2: To a stirred solution of methyl 3-(4-methyl-1,3-oxazol-2-yl)benzoate (180.00 mg; 0.829 mmol) in THF (2.0 mL) and water (1.0 mL), LiOH (39.7 mg; 1.658 mmol) was added, and the resulting mixture was stirred at room temperature for 4 h. The mixture was acidified to pH 1 with 2N HCl solution and extracted with dichloromethane. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / MeOH - 92:8) to yield 120 mg (71%) of 3-(4-methyl-1,3-oxazol-2-yl)benzoic acid as a colorless solid. HPLC / MS m / z: 204.1 [M+H] + , Rt(E): 0.76 min.

[0177] A4: 3-(1-chloro-7-isoquinolyl)-5-methyl-1,2,4-oxadiazole [ka]

[0178] A4.1: Nitrogen gas was bubbled through a mixture of 7-bromoisoquinolin-1-ol (0.500 g, 2.23 mmol) and zinc cyanide (0.341 g, 2.90 mmol) in DMF (12.4 mL) for 15 min. Palladium tetrakis(triphenylphosphine) (0.155 g, 0.13 mmol) was added, and the mixture was heated at 100 °C in a sealed vial for 16 h. The reaction mixture was diluted with brine (100 mL) and extracted with ethyl acetate (100 mL). The organic layer was washed with brine (2 x 50 mL). The aqueous layer, which contained some precipitated product, was re-extracted with dichloromethane (2 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, pre-absorbed onto silica, and purified by column chromatography (eluent: methanol / dichloromethane - 0-10% gradient) to yield 0.314 g (83%) of 1-hydroxyisoquinoline-7-carbonitrile. HPLC / MS m / z: 171.05 [M+H] + , Rt(AC): 0.84 min. 1 H NMR(500 MHz, DMSO-d6): δ 11.64(s, 1H), 8.53-8.47(m, 1H), 8.03(dd, J = 8.3, 1.8 Hz, 1H), 7.84(d, J = 8.3 Hz, 1H), 7.38(d, J = 7.1 Hz, 1H), 6.65(d, J = 7.2 Hz, 1H).

[0179] A4.2: 1-Hydroxyisoquinoline-7-carbonitrile (0.215 g, 1.26 mmol) and hydroxylamine hydrochloride (0.176 g, 2.52 mmol) in 1-butyl-3-methylimidazolium acetate (1.3 mL) were heated at 80° C. for 30 min. Water (50 mL) was added and the resulting precipitate was filtered, washed with water (50 mL) and dried to give 0.235 g (92%) of N′,1-dihydroxyisoquinoline-7-carboxamidine. HPLC / MS m / z: 204.07 [M+H] + , Rt(AC): 0.36 min.

[0180] A4.3.: N',1-Dihydroxyisoquinoline-7-carboxamidine (0.235 g, 1.16 mmol) and acetic anhydride (0.13 mL, 1.39 mmol) in acetonitrile (4.6 mL) were heated at 180 °C for 10 min by microwave irradiation. Water (25 mL) was added and the resulting precipitate was filtered, washed with water (25 mL), diethyl ether (2 x 10 mL) and dried to give 0.180 g (68%) of 7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-ol. HPLC / MS m / z: 228.08 [M+H] + , Rt(AC): 1.07 min.

[0181] A4.4: 7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-ol (50 mg, 0.22 mmol) in phosphorus oxychloride (2 mL) was heated at 100° C. for 1 h. The mixture was concentrated to give 0.054 g (100%) of 3-(1-chloro-7-isoquinolyl)-5-methyl-1,2,4-oxadiazole. HPLC / MS m / z: 246 / 248 Cl split [M+H] + , Rt(AC): 1.32 min.

[0182] A5: 5-methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [ka]

[0183] A5.1: Isoquinoline-7-carbonitrile (4.00 g, 25.9 mmol), triethylamine (7.23 mL, 51.9 mmol), and [bmim]OAc (26 mL) were mixed and heated to 80° C. Hydroxylamine hydrochloride (3.61 g, 51.9 mmol) was added. The reaction mixture was continued to stir at 80° C. for 1.5 h. The reaction mixture was cooled to room temperature and thoroughly mixed with EtOAc (250 mL). The emulsion was then mixed with water (750 mL). The layers were separated, and the aqueous layer was further extracted with EtOAc (5 x 100 mL). The combined organic layers were washed with saturated NaCl (3 x 100 mL), dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure to afford 3.70 g (76%) of N'-hydroxyisoquinoline-7-carboxamidine as an off-white solid. HPLC / MS m / z: 188.0829 [M+H] + , Rt(AD): 0.37 min.

[0184] A5.2: Four 20 mL microwave vials were each charged with one-quarter of the following: N'-hydroxyisoquinoline-7-carboxamidine (3.67 g, 19.6 mmol) was mixed with acetonitrile (40 mL) and acetic anhydride (2.2 mL, 23.5 mmol) under an argon atmosphere. Portions of the reaction mixture were each heated at 180 °C for 10 min under microwave irradiation. The combined reaction mixtures were evaporated onto silica gel and purified by flash chromatography (20-80% ethyl acetate in cyclohexane) to yield 3.59 g (87%) of 3-(7-isoquinolyl)-5-methyl-1,2,4-oxadiazole as an off-white solid. HPLC / MS m / z: 212.1 [M+H] + , Rt(AC): 0.88 min.

[0185] A5.3: 3-(7-Isoquinolyl)-5-methyl-1,2,4-oxadiazole (3.59 g, 17.0 mmol) was suspended in anhydrous chloroform (57 mL) under an argon atmosphere and cooled in an ice bath. 3-Chloroperoxybenzoic acid (4.57 g, 20.4 mmol) was added. The stirred reaction mixture was allowed to warm to ambient temperature and continued stirring overnight. Potassium carbonate (9.40 g, 68.0 mmol) was added. The mixture was stirred at room temperature for 4 h before being filtered through a pad of anhydrous MgSO4. The filtrate was concentrated under reduced pressure to afford 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole (3.12 g, 81%) as an off-white powder. HPLC / MS m / z: 228.0713 [M+H] + , Rt(AH): 1.73 min.

[0186] B1: Ethyl 2-(2-aminoethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate dihydrochloride [ka]

[0187] B1.1: To a stirred solution of methyl 5-bromo-2-(bromomethyl)benzoate (28.5 g; 92.542 mmol) and tert-butyl N-(2-aminoethyl)carbamate (62.3 g; 388.676 mmol) in MeOH (400.0 mL) was added triethylamine (18.7 g; 185.08 mmol) at room temperature, and the resulting mixture was stirred at 40 °C for 16 h. The resulting mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 1:1) to produce 32 g (96%) of tert-butyl N-[2-(6-bromo-1-oxo-2,3-dihydro-1H-isoindol-2-yl)ethyl]carbamate as an off-white solid. HPLC / MS m / z: 355.1-357.1 [M+H] + , Rt(A): 0.84 min.

[0188] B1.2: To a stirred solution of tert-butyl N-[2-(6-bromo-1-oxo-2,3-dihydro-1H-isoindol-2-yl)ethyl]carbamate (1.59 g, 4.476 mmol) and triethylamine (930.7 μL, 6.714 mmol) in MeOH (18.0 mL) and THF (52.0 mL) in a pressure tank, Pd(dppf)Cl.CHCl (218.4 mg, 0.273 mmol) and 1,1-bis-(diphenylphosphino)-ferrocene (198.5 mg, 0.358 mmol) were added. The mixture was purged with nitrogen for 3 min and then pressurized to 4.1 bar with carbon monoxide at 100 °C overnight. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The resulting mixture was concentrated in vacuo. The residue was purified by column chromatography to give 1.32 g (88%) of methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate as a red foam. HPLC / MS m / z: 234.9 [M+H-Boc] + , Rt(G): 1.38 min.

[0189] B1.3: Methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate (469.0 mg, 1.403 mmol)) was suspended in dioxane (4.8 mL) and a solution of HCl in dioxane (4.0 M, 2.1 mL) was added. A clear solution formed which became a suspension after 5 min. The reaction was stirred at room temperature overnight. A solution of HCl in dioxane (4.0 M, 0.7 mL) was added and the reaction mixture was stirred at room temperature overnight and evaporated to dryness to give 422.0 mg (98%) of methyl 2-(2-aminoethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate dihydrochloride as a pale pink solid. HPLC / MS m / z: 234.9 [M+H] + , Rt(G): 0.96 min.

[0190] B2: 2-(2-aminoethyl)-6-bromo-3H-isoindol-1-one [ka] To a stirred solution of methyl 5-bromo-2-(bromomethyl)benzoate (22.8 g, 74.034 mmol) and ethane-1,2-diamine (22.3 g, 370.906 mmol) in methanol (26.00 mL) at room temperature, triethylamine (12.4 g, 123.024 mmol) was added, and the resulting mixture was stirred at 40 °C for 4 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: chloroform / methanol - 75:25) to afford 15 g (79%) of 2-(2-aminoethyl)-6-bromo-3H-isoindol-1-one as an off-white solid. HPLC / MS m / z: 255.05 [M+H] + , Rt(J): 0.47 min.

[0191] B3: N1-[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]ethane-1,2-diamine [ka] 1-Chloro-7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinoline [Intermediate A4] (900.0 mg, 3.664 mmol) and ethylenediamine (3.71 mL, 54.953 mmol) were suspended in dry 1-methyl-2-pyrrolidinone (16 mL) in a 5 mL microwave bath, and the mixture was heated at 160 °C for 45 min under microwave irradiation. The reaction mixture was diluted with saturated aqueous NaHCO solution and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. After adding ethyl acetate and n-heptane to the residue, a precipitate formed, which was suction filtered, rinsed with n-heptane, and dried under high vacuum to give 726 mg (74%) of N1-[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]ethane-1,2-diamine as a pale yellow solid. HPLC / MS m / z: 270.0 [M+H] + , Rt (G): 0.90 min.

[0192] Example 1: Methyl 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] Intermediate B1 (422.0 mg, 1.374 mmol) and N-ethyldiisopropylamine (532.7 mg, 4.122 mmol) were dissolved in DMF (5.8 mL). 3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)benzoic acid (280.6 mg, 1.374 mmol) and [dimethylamino-([1,2,3]triazolo[4,5-b]pyridin-3-yloxy)-methylene]-dimethyl-ammonium; hexafluorophosphate (574.7 mg, 1.511 mmol) were added, and the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with a mixture of water and saturated aqueous NaHCO3 solution (1:2, 60 mL) and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to yield 577 mg (100%) as a pale red solid. HPLC / MS m / z: 421.9 [M+H] + , Rt (G): 1.33 min. 1 H NMR (700 MHz, DMSO-d6): δ 8.86 (t, J = 5.9 Hz, 1H), 8.45 (t, J = 1.8 Hz, 1H), 8.18-8.17 (m, 1H), 8.17-8.16 (m, 1H), 8.14-8.13 (m, 1H), 7.93-7.91 (m, 1H), 7.79-7.77 (m, 1H), 7.64 (t, J = 7.7 Hz, 1H), 4.66 (s, 2H), 4.44 (s, 3H), 3.88 (s, 3H), 3.76 (t, J = 6.0 Hz, 2H), 3.61-3.57 (m, 2H).

[0193] Example 2: Ethyl 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] Methyl 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate (73.6 mg, 0.175 mmol) was suspended in dry ethanol (4.90 mL). A solution of HCl in dioxane (4.0 M, 1.09 mL, 4.377 mmol) was added. A clear solution was formed, which was stirred at 55° C. for 60 h, cooled to room temperature, and evaporated to dryness. The residue was purified by chromatography to give 56 mg (74%) as a colorless solid. HPLC / MS m / z: 434.9 [M+H] + , Rt (G): 1.42 min. 1 H NMR (700 MHz, DMSO-d6): δ 8.86 (t, J = 5.9 Hz, 1H), 8.45 (t, J = 1.8 Hz, 1H), 8.18-8.16 (m, 2H), 8.14-8.13 (m, 1H), 7.93-7.91 (m, 1H), 7.79-7.77 (m, 1H), 7.64 (t, J = 7.7 Hz, 1H), 4.66 (s, 2H), 4.44 (s, 3H), 4.33 (q, J = 7.1 Hz, 2H), 3.76 (t, J = 6.0 Hz, 2H), 3.59 (q, J = 5.9 Hz, 2H), 1.34 (t, J = 7.1 Hz, 3H).

[0194] Example 3: Propan-2-yl 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka]

[0195] Example 3.1: Methyl 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate (137.9 mg, 0.328 mmol) was dissolved in THF (6.60 mL) and water (3.30 mL). Lithium hydroxide (19.7 mg, 0.821 mmol) was added and the reaction mixture was stirred at room temperature for 2 h. The THF was removed in vacuo and the aqueous residue was diluted with water (20 mL) and acidified to pH 3-4 with aqueous HCl solution. A solid formed which was filtered off with suction, washed with demineralized water and dried under vacuum to give 116 mg (86%) of 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylic acid as a pale beige solid. HPLC / MS m / z: 407.9 [M+H] + , Rt (H): 0.79 min.

[0196] Example 3.2: 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylic acid (115.0 mg, 0.283 mmol), 4-(dimethylamino)-pyridine (13.8 mg, 0.113 mmol), and DCC (93.3 mg, 0.452 mmol) were dissolved in dry THF (0.7 mL) and dry DMF (0.4 mL). 2-Propanol (216 μL, 2.826 mmol) was added and the reaction was stirred at room temperature overnight. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to give 88 mg (69%) of the title compound as a colorless solid. HPLC / MS m / z: 449.9 [M+H] + , Rt (G): 1.49 min. 1H NMR (700 MHz, DMSO-d6): δ 8.86 (t, J = 5.9 Hz, 1H), 8.45 (t, J = 1.7 Hz, 1H), 8.18-8.16 (m, 1H), 8.15 (dd, J = 7.9, 1.6 Hz, 1H), 8.13-8.12 (m, 1H), 7.93-7.91 (m, 1H), 7.78-7.76 (m, 1H), 7.64 (t, J = 7.7 Hz, 1H), 5.15 (quint, J = 6.3 Hz, 1H), 4.66 (s, 2H), 4.44 (s, 3H), 3.76 (t, J = 5.9 Hz, 2H), 3.59 (q, J = 5.9 Hz, 2H), 1.33 (d, J = 6.2 Hz, 6H).

[0197] Example 4: tert-Butyl 2-(2-{[3-(2-methyl-2H-1,2,3,4-tetrazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] A mixture of intermediate 3.1 (37.8 mg, 0.093 mmol) and O-(tert-butyl)-N,N'-diisopropylisourea (93.7 mg, 0.465 mmol) in dry toluene (0.7 mL) was stirred at 90 °C for 16 h. The reaction mixture was cooled to room temperature, diluted with water, and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by RP flash chromatography to yield 28 mg (65%) of the title compound as a colorless solid. HPLC / MS m / z: 463.9 [M+H] + , Rt (H): 0.93 min. 1H NMR (700 MHz, DMSO-d6): δ 8.86 (t, J = 5.8 Hz, 1H), 8.46 (t, J = 1.8 Hz, 1H), 8.19-8.16 (m, 1H), 8.11 (dd, J = 7.9, 1.6 Hz, 1H), 8.08-8.07 (m, 1H), 7.93-7.91 (m, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.64 (t, J = 7.8 Hz, 1H), 4.65 (s, 2H), 4.44 (s, 3H), 3.76 (t, J = 6.0 Hz, 2H), 3.59 (q, J = 5.9 Hz, 2H), 1.56 (s, 9H).

[0198] Example 5: Ethyl 2-(2-{[3-(3-methyl-1,2-oxazol-5-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] To a solution of Intermediate A1 (79.8 mg; 0.393 mmol) and Intermediate B1 (97.5 mg; 0.393 mmol) in DMF (2.0 mL), 1-methyl-1H-imidazole (64.5 mg; 0.785 mmol) and TCFH (165.3 mg; 0.589 mmol) were added, and the mixture was stirred at room temperature for 2 h. The reaction mixture was evaporated, and the residue was purified by RP flash chromatography to give 77 mg (45%) of the title compound as an off-white solid. HPLC / MS m / z: 434.15 [M+H] + , Rt (D): 1.09 min. 1H NMR (300 MHz, DMSO-d6): δ 8.69 (t, J = 5.8 Hz, 1H), 8.10-7.99 (m, 3H), 7.83 (dt, J = 7.8, 1.4 Hz, 1H), 7.73 (dt, J = 7.9, 1.4 Hz, 1H), 7.65 (d, J = 7.9 Hz, 1H), 7.48 (t, J = 7.8 Hz, 1H), 6.78 (s, 1H), 4.54 (s, 2H), 4.28-4.15 (m, 2H), 3.68-3.58 (m, 2H), 3.52-3.41 (m, 2H), 2.18 (s, 3H), 1.22 (t, J = 7.1 Hz, 3H).

[0199] Example 6: Ethyl 2-(2-{[3-(5-methyl-1,3-oxazol-2-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] Prepared as described in Example 5 using A2 (100.00 mg; 0.49 mmol) and B1 (122.1 mg; 0.49 mmol). Purification by reversed-phase flash chromatography. Yield: 24 mg (11%) of a colorless solid. HPLC / MS m / z: 434.15 [M+H] + , Rt (F): 1.55 min. 1H NMR (400 MHz, DMSO-d6): δ 8.83 (t, J = 5.8 Hz, 1H), 8.30 (t, J = 1.8 Hz, 1H), 8.20-8.12 (m, 2H), 8.04 (dt, J = 7.8, 1.4 Hz, 1H), 7.86 (dt, J = 7.9, 1.4 Hz, 1H), 7.81-7.74 (m, 1H), 7.59 (t, J = 7.8 Hz, 1H), 7.02 (t, J = 1.3 Hz, 1H), 4.66 (s, 2H), 4.38-4.29 (m, 2H), 3.79-3.72 (m, 2H), 3.63-3.54 (m, 2H), 2.39 (d, J = 1.3 Hz, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0200] Example 7: Ethyl 2-(2-{[3-(4-methyl-1,3-oxazol-2-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] Prepared as described in Example 5 using A3 (100.0 mg; 0.49 mmol) and B1 (122.2 mg; 0.49 mmol). Purification by reversed-phase flash chromatography. Yield: 48 mg (23%) of a colorless solid. HPLC / MS m / z: 434.15 [M+H] + , Rt (C): 0.90 min. 1H NMR (400 MHz, DMSO-d6): δ 8.84 (t, J = 5.9 Hz, 1H), 8.33 (t, J = 1.8 Hz, 1H), 8.20-8.11 (m, 2H), 8.05 (dt, J = 7.8, 1.4 Hz, 1H), 7.93 (q, J = 1.2 Hz, 1H), 7.88 (dt, J = 7.8, 1.4 Hz, 1H), 7.77 (dd, J = 7.9, 0.8 Hz, 1H), 7.59 (t, J = 7.8 Hz, 1H), 4.66 (s, 2H), 4.39-4.29 (m, 2H), 3.79-3.72 (m, 2H), 3.63-3.54 (m, 2H), 2.17 (d, J = 1.3 Hz, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0201] Example 8: Ethyl 2-(2-{[3-(5-methyl-1,2,4-oxadiazol-3-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka]

[0202] Example 8.1. To a stirred solution of intermediate B2 (599.3 mg; 2.349 mmol) and 3-(5-methyl-1,2,4-oxadiazol-3-yl)benzoic acid (575.6 mg; 2.819 mmol) in DMF (6.0 mL) was added 1-methyl-1H-imidazole (578.6 mg; 7.047 mmol) at room temperature. After stirring the mixture for 15 min at room temperature, [chloro(dimethylamino)methylidene]dimethylazanium; hexafluoro-λ 5N-phosphanide (791.0 mg; 2.819 mmol) was added at room temperature, and the mixture was stirred for 2 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 9:1) to give 666 mg (64%) of N-[2-(6-bromo-1-oxo-2,3-dihydro-1H-isoindol-2-yl)ethyl]-3-(5-methyl-1,2,4-oxadiazol-3-yl)benzamide as a colorless solid. HPLC / MS m / z: 441.05 [M+H] + , Rt (J): 0.66 min.

[0203] Example 8.2. To a solution of intermediate 8.1 (174.0 mg, 0.394 mmol) and triethylamine (190.00 μL, 1.299 mmol) in EtOH (16.0 mL) was added Pd(dppf)Cl2.CHCl2 (35.1 mg, 0.043 mmol) in a pressure tank. The mixture was purged with nitrogen for 3 min and then pressurized to 30 atm with carbon monoxide at 100 °C for 32 h. The reaction mixture was cooled to room temperature, filtered to remove insoluble solids, and the filtrate was concentrated under vacuum. The crude product was purified by preparative HPLC to give 29 mg (17%) of ethyl 2-(2-{[3-(5-methyl-1,2,4-oxadiazol-3-yl)phenyl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate as a colorless solid. HPLC / MS m / z: 435.20 [M+H] + , Rt (I): 1.85 min. 1H NMR (300 MHz, DMSO-d6): δ 8.87 (t, J = 5.8 Hz, 1H), 8.39 (t, J = 1.7 Hz, 1H), 8.22-8.07 (m, 3H), 8.01-7.92 (m, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.65 (t, J = 7.8 Hz, 1H), 4.66 (s, 2H), 4.34 (q, J = 7.1 Hz, 2H), 3.76 (t, J = 5.9 Hz, 2H), 3.59 (q, J = 5.9 Hz, 2H), 2.68 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0204] The following examples were prepared in a similar manner: Example 9: Ethyl 2-(2-{[6-(5-methyl-1,2,4-oxadiazol-3-yl)pyridin-2-yl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] 36 mg of colorless solid. HPLC / MS m / z: 436.9 [M+H] + , Rt (G): 1.42 min. 1 H NMR (700 MHz, DMSO-d6): δ 8.83 (t, J = 6.3 Hz, 1H), 8.21 (dd, J = 7.7, 1.3 Hz, 1H), 8.19-8.17 (m, 1H), 8.17-8.16 (m, 1H), 8.13 (dd, J = 7.7, 1.3 Hz, 1H), 8.12-8.11 (m, 1H), 7.78-7.76 (m, 1H), 4.69 (s, 2H), 4.33 (q, J = 7.1 Hz, 2H), 3.79 (t, J = 5.9 Hz, 2H), 3.65 (q, J = 6.1 Hz, 2H), 2.70 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0205] Example 10: Propan-2-yl 2-(2-{[6-(5-methyl-1,2,4-oxadiazol-3-yl)pyridin-2-yl]formamido}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] 60 mg of colorless solid. HPLC / MS m / z: 450.9 [M+H] + , Rt (G): 1.49 min. 1 H NMR (700 MHz, DMSO-d6): δ 8.82 (t, J = 6.2 Hz, 1H), 8.21 (dd, J = 7.7, 1.3 Hz, 1H), 8.18 (t, J = 7.7 Hz, 1H), 8.15 (dd, J = 7.9, 1.6 Hz, 1H), 8.13 (dd, J = 7.7, 1.3 Hz, 1H), 8.11-8.10 (m, 1H), 7.77-7.75 (m, 1H), 5.18-5.12 (m, 1H), 4.69 (s, 2H), 3.79 (t, J = 5.9 Hz, 2H), 3.65 (q, J = 6.1 Hz, 2H), 2.70 (s, 3H), 1.33 (d, J = 6.2 Hz, 6H).

[0206] Example 11: Ethyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka]

[0207] Example 11.1: To a solution of intermediate B2 (1.00 g, 3.920 mmol) and 1-chloro-7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinoline (0.96 g, 3.920 mmol) in NMP (8.0 mL) at room temperature was added diethylamine (1.45 g, 11.181 mmol). The resulting mixture was heated to 160° C. and stirred for 16 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by RP flash chromatography to give 1.2 g (66%) of 6-bromo-2-(2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino]ethyl)-3H-isoindol-1-one as a yellow solid. HPLC / MS m / z: 466.10 [M+H] + , Rt (J): 0.62 min.

[0208] Example 11.2: To a solution of 6-bromo-2-(2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino]ethyl)-3H-isoindol-1-one (48.8 mg, 0.105 mmol) in ethanol (6.0 mL) was added Pd(dppf)Cl2.CHCl2 (10.6 mg, 0.013 mmol), triethylamine (28.5 mg, 0.282 mmol) in a pressure tank. The mixture was purged with nitrogen for 1 min and then pressurized to 30 atm with carbon monoxide at 100 °C for 16 h. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The resulting mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 1:1) and preparative HPLC to give 18 mg (38%) of the title compound as a colorless solid. HPLC / MS m / z: 458.0 [M+H]+, Rt (K): 0.93 min. 1H NMR (300 MHz, DMSO-d6): δ 8.83 (s, 1H), 8.19-8.12 (m, 3H), 8.04 (d, J = 5.6 Hz, 1H), 7.94 (d, J = 5.7 Hz, 1H), 7.85 (d, J = 8.6 Hz, 1H), 7.76 (d, J = 7.9 Hz, 1H), 6.95 (d, J = 5.8 Hz, 1H), 4.69 (s, 2H), 4.34 (q, J = 7.1 Hz, 2H), 3.92-3.74 (m, 4H), 2.68 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0209] Example 12: Ethyl 2-(2-{[7-(1-methyl-1H-pyrazol-4-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] Prepared in a similar manner. 28 mg off-white solid. HPLC / MS m / z: 456.15 [M+H] +, Rt (L): 0.59 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.33 (s, 1H), 8.19-8.10 (m, 3H), 7.95 (s, 1H), 7.84-7.70 (m, 3H), 7.66 (d, J = 8.5 Hz, 1H), 7.58 (s, 1H), 6.83 (d, J = 5.8 Hz, 1H), 4.68 (s, 2H), 4.32 (q, J = 7.1 Hz, 2H), 3.92-3.73 (m, 7H), 1.32 (t, J = 7.1 Hz, 3H).

[0210] Example 13: Propan-2-yl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka]

[0211] Example 13.1: To a stirred solution of Intermediate 11.1 (480.0 mg, 1.034 mmol) and formyl acetate (0.85 mL, 10.269 mmol) in DMF (6.0 mL) was added triethylamine (0.76 mL, 5.18 mmol) and Pd(dppf)Cl.CHCl (64.5 mg, 0.079 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was stirred at 90 °C under a nitrogen atmosphere for 2 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by RP flash chromatography to give 292 mg (66%) of 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylic acid as a yellow solid. HPLC / MS m / z: 430.15 [M+H] + , Rt (J): 0.57 min.

[0212] Example 13.2: A solution of intermediate 12.1 (87.6 mg, 0.204 mmol) in HSO (0.25 mL, 4.660 mmol) and 2-propanol (1.0 mL) was stirred at 80° C. for 3 h. The mixture was cooled to room temperature, basified to pH 8 with saturated aqueous NaHCO solution, and extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by preparative HPLC to afford 32 mg (33%) of the title compound as a colorless solid. HPLC / MS m / z: 472.10 [M+H] + , Rt(B): 0.76 min. 1 H NMR(400 MHz, DMSO-d6): δ 8.82 (s, 1H), 8.18-8.08 (m, 3H), 8.03 (t, J = 5.6 Hz, 1H), 7.93 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.75 (d, J = 7.8 Hz, 1H), 6.94 (d, J = 5.7 Hz, 1H), 5.22-5.08 (m, 1H), 4.68 (s, 2H), 3.87 (t, J = 5.7 Hz, 2H), 3.81 (t, J = 5.5 Hz, 2H), 2.67 (s, 3H), 1.33 (d, J = 6.3 Hz, 6H).

[0213] Example 14: tert-Butyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-isoindole-5-carboxylate [ka] To O-(tert-butyl)-N,N'-diisopropylisourea (1.0 mL) was added Intermediate 12.1 (87.6 mg, 0.204 mmol), and the mixture was stirred at 60 °C for 3 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The crude product was purified by preparative HPLC to give 23 mg (23%) of the title compound as a colorless solid. HPLC / MS m / z: 472.10 [M+H] + , Rt (B): 0.79 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.82 (d, J = 1.5 Hz, 1H), 8.17-8.08 (m, 2H), 8.06-8.01 (m, 2H), 7.93 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.73 (d, J = 7.9 Hz, 1H), 6.94 (d, J = 5.7 Hz, 1H), 4.67 (s, 2H), 3.86 (t, J = 5.7 Hz, 2H), 3.83-3.77 (m, 2H), 2.67 (s, 3H), 1.55 (s, 9H).

[0214] Example 15: 6-(Difluoromethoxy)-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindol-1-one [ka]

[0215] Example 15.1: To a stirred suspension of methyl 2-(bromomethyl)-5-methoxybenzoate (200.0 mg, 0.772 mmol) and Intermediate B3 (228.7 mg, 0.849 mmol) in dry methanol (4.8 mL) was added triethylamine (321.0 μL, 2.316 mmol). The reaction mixture was heated to 45° C. to form a clear solution, which was stirred at this temperature for 90 min. The reaction mixture was cooled to room temperature and evaporated to dryness. The residue was purified by RP flash chromatography to give 137 mg (43%) of 6-methoxy-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindol-1-one as a beige solid. HPLC / MS m / z: 415.9 [M+H] + , Rt (H): 0.71 min.

[0216] Example 15.2: 6-Methoxy-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindol-1-one (124.0 mg, 0.293 mmol) was suspended in dry dichloromethane (4.7 mL). Boron tribromide (142.4 μL, 1.463 mmol) was dissolved in dry dichloromethane (1.9 mL) and added slowly to the suspension, and the mixture was stirred at room temperature for 4 h. The reaction mixture was poured into a 1:1 mixture of water / aqueous saturated NaHCO3 solution (60 mL) and extracted with dichloromethane. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by chromatography to give 63 mg (54%) of 6-hydroxy-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindol-1-one as a beige solid. HPLC / MS m / z: 401.9 [M+H] + , Rt (H): 0.67 min.

[0217] Example 15.3: Intermediate 15.2 (53.0 mg, 0.132 mmol) and cesium carbonate (64.5 mg, 0.198 mmol) were suspended in DMF (770.0 μL). Methyl 2-chloro-2,2-difluoroacetate (19.5 μL, 0.185 mmol) was added and the mixture was stirred at 80° C. for 3 h. Further methyl 2-chloro-2,2-difluoroacetate (14.0 μL, 0.132 mmol) and cesium carbonate (43.5 mg, 0.132 mmol) were added and the reaction was stirred at 80° C. overnight. The reaction mixture was cooled to room temperature, filtered, and evaporated to dryness. The residue was purified by RP flash chromatography to give 21 mg (35%) of the title compound as a colorless solid. HPLC / MS m / z: 451.8 [M+H] + , Rt (M): 1.78 min. 1 H NMR (700 MHz, DMSO-d6): δ 8.84-8.82 (m, 1H), 8.14 (dd, J = 8.4, 1.5 Hz, 1H), 8.03 (t, J = 5.6 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.65 (d, J = 8.1 Hz, 1H), 7.40 (d, J = 2.4 Hz, 1H), 7.38 (dd, J = 8.2, 2.4 Hz, 1H), 7.32 (t, J = 73.9 Hz, 1H), 6.95 (d, J = 5.7 Hz, 1H), 4.58 (s, 2H), 3.87-3.84 (m, 2H), 3.81-3.78 (m, 2H), 2.68 (s, 3H).

[0218] Example 16: 6-(Difluoromethoxy)-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindol-1-one [ka]

[0219] Example 16.1: Methyl 2-bromo-5-(trifluoromethoxy)benzoate (1.43 g, 4.782 mmol) and zinc cyanide (2.28 g, 19.417 mmol) were dissolved in dry DMF (22.0 mL) under a nitrogen atmosphere. Tetrakis(triphenylphosphine)palladium(0) (551.2 mg; 0.477 mmol) was added, and the reaction mixture was heated to 120° C. and stirred for 1 h. After cooling to room temperature, the reaction mixture was diluted with water and extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over sodium sulfate, filtered, and concentrated under vacuum. The crude product was purified by flash chromatography to yield 951 mg (81%) of methyl 2-cyano-5-(trifluoromethoxy)benzoate as a colorless solid. HPLC / MS m / z: 245.9 [M+H] + , Rt (G): 1.59 min.

[0220] Example 16.2: To a solution of methyl 2-cyano-5-(trifluoromethoxy)benzoate (742.0 mg, 3.027 mmol) in methanol (10.0 mL) was added Pd / C (5%, 800 mg) and HCl solution (0.76 mL, 32%) in a pressure tank. The mixture was hydrogenated under 3.6 bar hydrogen pressure at room temperature overnight, filtered, and evaporated to dryness. The residue (824 mg, 95%) was used in the next step without further purification.

[0221] Example 16.3: Intermediate 16.2 (824.0 mg, 2.885 mmol) was dissolved in methanol (25.0 mL) and treated with aqueous NaOH solution (7N, 5 mL). The reaction was stirred at room temperature for 20 min. The reaction mixture was concentrated under reduced pressure, and the aqueous residue was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and evaporated to dryness to give 571 mg (91%) of 6-(trifluoromethoxy)-2,3-dihydro-1H-isoindol-1-one as colorless crystals. HPLC / MS m / z: 217.9 [M+H] + , Rt (G): 1.32 min.

[0222] Example 16.4: Intermediate 16.3 (97.0 mg, 0.447 mmol) was dissolved in THF (3.6 mL) and treated with sodium hydride (60% dispersion in mineral oil; 53.6 mg, 1.340 mmol) under a nitrogen atmosphere while cooling in an ice bath. The suspension was stirred for 45 min. Bromoacetonitrile (214.3 mg, 1.787 mmol) was added, and the reaction mixture was allowed to warm to room temperature and stirred for 15 min. The reaction was quenched by the addition of water, and the mixture was extracted with ethyl acetate. The combined extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by RP flash chromatography to yield 65 mg (57%) of 2-[1-oxo-6-(trifluoromethoxy)-2,3-dihydro-1H-isoindol-2-yl]acetonitrile as an orange solid. HPLC / MS m / z: 256.9 [M+H] + , Rt (G): 1.42 min.

[0223] Example 16.5: To a solution of intermediate 16.4 (65 mg, 0.254 mmol) in methanol (3.0 mL) was added Pd / C (5%, 100 mg) and HCl solution (64 μL, 32%) in a pressure tank. The mixture was hydrogenated under 3.2 bar hydrogen pressure at room temperature for 17 h, filtered and evaporated to dryness. The crude product was purified by RP flash chromatography to give 24 mg (31%) of ammonium 2-[1-oxo-6-(trifluoromethoxy)-2,3-dihydro-1H-isoindol-2-yl]ethane-1-formate as a colorless solid. HPLC / MS m / z: 260.9 [M+H] + , Rt (G): 1.16 min.

[0224] Example 16.6: 7-(5-Methyl-1,2,4-oxadiazol-3-yl)isoquinolin-2-ium-2-olate (17.0 mg, 0.075 mmol) and Intermediate 16.5 (24.1 mg, 0.079 mmol) were dissolved in dry dichloromethane (0.4 mL) under an argon atmosphere. DIPEA (85.9 μL, 0.505 mmol) and bromotripyrrolidinophosphonium hexafluorophosphate (52.2 mg, 0.112 mmol) were added, and the reaction mixture was stirred at room temperature for 3 d. The reaction mixture was evaporated under reduced pressure and the crude product was purified by RP flash chromatography to give 11 mg (31%) of 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-6-(trifluoromethoxy)-2,3-dihydro-1H-isoindol-1-one as a colorless solid. HPLC / MS m / z: 470.8 [M+H] + , Rt (M): 1.87 min.

[0225] Example 17: 6-(2,3-difluoropropoxy)-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindol-1-one [ka]

[0226] Example 17.1: Methyl 5-hydroxy-2-methylbenzoate (122.5 mg, 0.737 mmol), 2,3-difluoropropan-1-ol (141.6 mg, 1.474 mmol), and triphenylphosphine (271.0 mg, 1.033 mmol) were dissolved in dry dichloromethane (2.2 mL) and the solution was cooled to 0° C. Di-tert-butyl azodicarboxylate (238.0 mg, 1.033 mmol) was added, and the pale yellow solution was stirred at 0° C. for 10 min and then at room temperature overnight. Triphenylphosphine (97.0 mg, 0.370 mmol) and di-tert-butyl azodicarboxylate (85.2 mg, 0.370 mmol) were added, and the mixture was heated to 40° C. and stirred for 4.5 h. The reaction mixture was diluted with dichloromethane, washed with water, dried over sodium sulfate, filtered, and evaporated to dryness. The oily residue was purified by flash chromatography to afford 170 mg (94%) of methyl 5-(2,3-difluoropropoxy)-2-methylbenzoate as a pale yellow oil. HPLC / MS m / z: 245.1 [M+H] + , Rt (N): 1.60 min.

[0227] Example 17.2: Intermediate 16.1 (205.0 mg, 0.839 mmol), NBS (149.3 mg, 0.839 mmol), and dibenzoyl peroxide (25% aqueous, 13.8 mg, 0.042 mmol) were suspended in carbon tetrachloride (7.0 mL) under argon, and the mixture was heated to 77 °C for 4 h. Further dibenzoyl peroxide (25% aqueous, 16.0 mg, 0.049 mmol) was added, and the reaction mixture was stirred at 77 °C overnight. The reaction mixture was cooled to room temperature, diluted with dichloromethane (20 mL), washed with water, dried over sodium sulfate, filtered, and evaporated to dryness. The residue was purified by flash chromatography to yield 222 mg (82%) of methyl 2-(bromomethyl)-5-(2,3-difluoropropoxy)benzoate as a colorless oil. HPLC / MS m / z: 345.0 [M+Na] + , Rt (N): 1.65 min.

[0228] Example 17.3. The reaction of Intermediate B3 (48.5 mg, 0.180 mmol) with Intermediate 17.2 (70.0 mg, 0.217 mmol) was carried out as described in Intermediate 15.1. Yield: 42 mg (49%) of a light brown solid. HPLC / MS m / z: 480.2 [M+H] + , Rt (N): 1.38 min. 1 H NMR (700 MHz, DMSO-d6): δ 8.85-8.83 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.04 (t, J = 5.6 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.51-7.49 (m, 1H), 7.19 (s, 1H), 7.19-7.17 (m, 1H), 6.95 (d, J = 5.6 Hz, 1H), 5.20-5.07 (m, 1H), 4.85-4.67 (m, 2H), 4.51 (s, 2H), 4.39-4.33 (m, 1H), 4.30-4.24 (m, 1H), 3.84 (t, J = 5.9 Hz, 2H), 3.78 (q, J = 5.8 Hz, 2H), 2.68 (s, 3H).

[0229] Example 18: 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-6-propoxyisoindolin-1-one [ka]

[0230] Example 18.1: To a mixture of 5-hydroxy-3-methylbenzoic acid methyl ester (200 mg, 1.2 mmol) and 1-iodopropane (0.24 mL, 2.4 mmol) in acetonitrile (3.3 mL) was added potassium carbonate (333 mg, 2.4 mmol) at rt. The resulting mixture was heated to reflux overnight. After 19 h, the reaction mixture was partitioned between ethyl acetate (60 mL) and water (40 mL). After phase separation, the organic layer was washed with brine (30 mL), dried over magnesium sulfate, filtered, and evaporated to dryness to yield methyl 2-methyl-5-propoxybenzoate (220 mg, 80%, 1.0 mmol), which was carried on to the next step without further purification. HPLC / MS m / z: (doesn't ionize), Rt (Y): 1.601 min.

[0231] Example 18.2: Methyl 2-methyl-5-propoxybenzoate (220 mg, 1.0 mmol) was dissolved in chloroform (1.5 mL). Azobisisobutyronitrile (AIBN) (2.6 mg, 0.14 mmol) and N-bromosuccinimide (207 mg, 1.2 mmol) were carefully added to the solution, and the resulting mixture was refluxed at 65 °C overnight. After 17 h of reaction, the reaction mixture was evaporated onto silica (dry loading) and purified by silica column chromatography (eluent: 0-30% ethyl acetate in cyclohexane) to give the desired product, methyl 2-(bromomethyl)-5-propoxybenzoate (230 mg, 76%, 0.8 mmol). HPLC / MS m / z: (doesn't ionize), Rt (Y): 1.625 min.

[0232] Example 18.3: A solution of methyl 2-(bromomethyl)-5-propoxybenzoate (230 mg, 0.8 mmol), 1-Boc-ethylenediamine (0.15 mL, 0.9 mmol), and triethylamine (0.17 mL, 1.2 mmol) in methanol (4 mL) was refluxed for 23 h. After cooling to room temperature, the reaction mixture was concentrated and purified by silica column chromatography (eluent: 20-90% ethyl acetate in cyclohexane) to afford tert-butyl N-[2-(1-oxo-6-propoxy-isoindolin-2-yl)ethyl]carbamate (140 mg, 53%, 0.4 mmol) as a white amorphous solid. HPLC / MS m / z: 357.2 [M+Na] + , Rt (Y): 1.493 min.

[0233] Example 18.4: tert-Butyl N-[2-(1-oxo-6-propoxyisoindolin-2-yl)ethyl]carbamate (140 mg, 0.4 mmol) was mixed with 4 M HCl in dioxane (10 mL, 42 mmol) and 1,4-dioxane (10 mL) and stirred for 18 h. Volatiles were removed under reduced pressure, and the resulting crude material (176 mg) was dissolved in methanol and subsequently purified using an SCX-II (2 g, 15 mL) cartridge with methanol, a 2 M ammonia solution in methanol as the eluent. The basic fractions were combined to give the product 2-(2-aminoethyl)-6-propoxyisoindolin-1-one (98 mg, 100%, 0.4 mmol) as a white amorphous powder. HPLC / MS m / z: 235.1 [M+H] + , Rt (Y): 0.946 min.

[0234] Example 18.5: 2-(2-aminoethyl)-6-propoxyisoindolin-1-one (40 mg, 0.17 mmol), 5-methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (46 mg, 0.2 mmol), PyBrop (96 mg, 0.2 mmol), N,N-diisopropylethylamine (0.11 mL, 0.6 mmol), and anhydrous dichloromethane (0.85 mL) were placed in a microwave vial under nitrogen at room temperature. The reaction mixture was heated at 60° C. by microwave irradiation for 1 h. The volatiles were removed under reduced pressure, and the resulting crude material was directly purified by reverse-phase flash chromatography (eluent: 20–100% MeOH in water modified with 0.1% formic acid), followed by purification using an SCX-II (2 g, 15 mL) cartridge with methanol, 2 M ammonia solution in methanol as eluent. The basic fractions were concentrated to dryness in vacuo to afford 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6-propoxyisoindolin-1-one (22 mg, 29%, 0.05 mmol) as a pale yellow solid. HPLC / MS m / z: 444.2 [M+H] + , Rt (Y): 1.287 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.84 (br s, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.03 (t, J = 5.5 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.46 (dd, J = 8.1, 0.9 Hz, 1H), 7.15-7.08 (m, 2H), 6.95 (dd, J = 5.9, 0.8 Hz, 1H), 4.49 (s, 2H), 3.96 (t, J = 6.6 Hz, 2H), 3.83 (t, J = 5.9 Hz, 2H), 3.78 (q, J = 5.6 Hz, 2H), 2.68 (s, 3H), 1.89-1.62 (m, 2H), 0.97 (t, J = 7.4 Hz, 3H).

[0235] Example 19: 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-6-(trifluoromethyl)-2,3-dihydro-1H-isoindol-1-one [ka]

[0236] Example 19.1: Prepared as described in Intermediate B2 using methyl 2-(bromomethyl)-5-(trifluoromethyl)benzoate (2.38 g, 8.012 mmol), ethane-1,2-diamine (2.41 g, 40.058 mmol), and triethylamine (2.43 g, 24.036 mmol) in methanol (26.00 mL). Yield: 1.69 g (86%) of 2-(2-aminoethyl)-6-(trifluoromethyl)-2,3-dihydro-1H-isoindol-1-one as a yellow solid. HPLC / MS m / z: 245.20 [M+H] + , Rt (B): 0.57 min.

[0237] Example 19.2: Prepared as described in Intermediate 11.1. Yield: 31 mg (9%) of 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-6-(trifluoromethyl)-2,3-dihydro-1H-isoindol-1-one as a yellow solid. HPLC / MS m / z: 454.00 [M+H] + , Rt (O): 1.54 min. 1H NMR (400 MHz, DMSO-d6): δ 8.82 (d, J = 1.6 Hz, 1H), 8.17-8.10 (m, 1H), 8.03 (t, J = 5.6 Hz, 1H), 7.99-7.90 (m, 2H), 7.90-7.81 (m, 3H), 6.97-6.91 (m, 1H), 4.70 (s, 2H), 3.92-3.84 (m, 2H), 3.82 (t, J = 5.6 Hz, 2H), 2.67 (s, 3H).

[0238] Example 20: 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-6-[1-(trifluoromethyl)cyclopropyl]-2,3-dihydro-1H-isoindol-1-one [ka]

[0239] Example 20.1: To a stirred solution of [3-(methoxycarbonyl)-4-methylphenyl]boronic acid (1.90 g, 9.794 mmol) and 2-bromo-3,3,3-trifluoroprop-1-ene (2.91 g, 16.633 mmol) in THF (14.0 mL) and water (7.0 mL) was added CsCO (4.47 g, 13.719 mmol) and Pd(dppf)Cl.CHCl (0.40 g, 0.490 mmol). The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 16 h. The resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 3:1) to give 1.89 g (79%) of methyl 2-methyl-5-(3,3,3-trifluoroprop-1-en-2-yl)benzoate as a yellow liquid. HPLC / MS m / z: 245.15 [M+H] + , Rt (J): 0.81 min.

[0240] Example 20.2: A stirred solution of intermediate 20.1 (1.79 g, 7.329 mmol) and diphenylmethylsulfonium tetrafluoroborate (2.74 g, 9.516 mmol) in THF (20.0 mL) was treated dropwise with sodium bis(trimethylsilyl)amide (2 M in THF, 5.86 mL, 11.711 mmol) at 0 °C under a nitrogen atmosphere. The resulting mixture was warmed to room temperature and stirred for 1 h. The reaction was quenched with water, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / petroleum ether - 1:2) to afford 851 mg (45%) of methyl 2-methyl-5-[1-(trifluoromethyl)cyclopropyl]benzoate as a yellow liquid. HPLC / MS m / z: 259.15 [M+H] + , Rt (B): 1.09 min.

[0241] Example 20.3: To a stirred solution of intermediate 20.2 (789.0 mg, 3.055 mmol) and NBS (598.0 mg, 3.360 mmol) in CCl4 (10.0 mL) was added AIBN (100.3 mg, 0.611 mmol). The resulting mixture was stirred at 70 °C for 2 h. The resulting mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether - 1:3) to afford 526 mg (51%) of methyl 2-(bromomethyl)-5-[1-(trifluoromethyl)cyclopropyl]benzoate as a yellow solid. HPLC / MS m / z: 339.05 [M+H] + , Rt (E): 1.14 min.

[0242] Example 20.4: Prepared as described in Intermediate 19.1. Yield: 149 mg (72%) of 2-(2-aminoethyl)-6-[1-(trifluoromethyl)cyclopropyl]-2,3-dihydro-1H-isoindol-1-one as a yellow solid. HPLC / MS m / z: 285.15 [M+H]+ , Rt(B): 0.68 min.

[0243] Example 20.5: To a stirred solution of Intermediate 20.4 (134.0 mg, 0.471 mmol) and 7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-2-ium-2-olate (107.1 mg, 0.471 mmol) in dichloromethane (3.0 mL) was added PyBrOP (285.5 mg, 0.612 mmol) and DIPEA (243.5 mg, 1.884 mmol). The resulting mixture was stirred at room temperature for 2 h and concentrated under reduced pressure. The residue was purified by RP flash chromatography to give 52 mg (22%) of 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-6-[1-(trifluoromethyl)cyclopropyl]-2,3-dihydro-1H-isoindol-1-one as a colorless solid. HPLC / MS m / z: 494.15 [M+H] + , Rt (F): 1.59 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.85-8.80 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.02 (t, J = 5.5 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.70-7.63 (m, 2H), 7.61 (dd, J = 7.6, 1.0 Hz, 1H), 6.98-6.92 (m, 1H), 4.58 (s, 2H), 3.89-3.83 (m, 2H), 3.83-3.75 (m, 2H), 2.67 (s, 3H), 1.41-1.33 (m, 2H), 1.16 (s, 2H).

[0244] Example 21: Methyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridine-3-carboxylate [ka]

[0245] Example 21.1: Ethyl 5-bromo-2-methylnicotinate (1.00 g, 4.097 mmol) was brominated with NMB in CCl4 as described in Intermediate 17.2. Yield: 855 mg (64%) of ethyl 5-bromo-2-(bromomethyl)pyridine-3-carboxylate as a colorless oil. HPLC / MS m / z: 323.7 [M+H] + , Rt (G): 1.72 min.

[0246] Example 21.2: Intermediate 21.1 (150.0 mg, 0.464 mmol) was treated with Intermediate B3 (160.0 mg, 0.594 mmol) and triethylamine (141.0 mg, 1.393 mmol) as described for Compound 17.2. Yield: 187 mg (87%) of 3-bromo-6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5H,6H,7H-pyrrolo[3,4-b]pyridin-5-one as an amorphous beige powder. HPLC / MS m / z: 465.1 / 467.0 [M+H] + , Rt (N): 1.28 min.

[0247] Example 21.3: Carbonylation of intermediate 21.2 to the title compound was carried out as described for compound 11.2. Yield: 31 mg (41%) of a beige solid. HPLC / MS m / z: 444.9 [M+H] + , Rt (G): 1.28 min. 1H NMR (400 MHz, DMSO-d6): δ 9.24 (d, J = 2.0 Hz, 1H), 8.81-8.79 (m, 1H), 8.34 (d, J = 2.0 Hz, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 7.98 (t, J = 5.4 Hz, 1H), 7.91 (d, J = 5.7 Hz, 1H), 7.83 (d, J = 8.5 Hz, 1H), 6.93 (d, J = 5.6 Hz, 1H), 4.76 (s, 2H), 3.91 (s, 3H), 3.91-3.87 (m, 2H), 3.87-3.81 (m, 2H), 2.67 (s, 3H).

[0248] The following examples were prepared in a similar manner: Example 22: Ethyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridine-3-carboxylate [ka] 20mg colorless solid. HPLC / MS m / z: 458.8 [M+H] + , Rt (G): 1.37 min. 1 H NMR (700 MHz, DMSO-d6): δ 9.24 (d, J = 2.0 Hz, 1H), 8.80-8.79 (m, 1H), 8.34 (d, J = 1.9 Hz, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.00 (t, J = 5.7 Hz, 1H), 7.91 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.94 (d, J = 5.7 Hz, 1H), 4.76 (s, 2H), 4.37 (q, J = 7.1 Hz, 2H), 3.91-3.88 (m, 2H), 3.86-3.82 (m, 2H), 2.67 (s, 3H), 1.35 (t, J = 7.1 Hz, 3H).

[0249] Example 23: Propan-2-yl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridine-3-carboxylate [ka] 31 mg of colorless solid. HPLC / MS m / z: 472.9 [M+H] + , Rt (M): 1.77 min. 1 H NMR (700 MHz, DMSO-d6): δ 9.22 (d, J = 2.0 Hz, 1H), 8.80-8.79 (m, 1H), 8.32 (d, J = 2.0 Hz, 1H), 8.14 (dd, J = 8.5, 1.6 Hz, 1H), 8.00 (t, J = 5.6 Hz, 1H), 7.91 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.93 (d, J = 5.9 Hz, 1H), 5.21-5.15 (m, 1H), 4.76 (s, 2H), 3.91-3.88 (m, 2H), 3.86-3.82 (m, 2H), 2.67 (s, 3H), 1.34 (d, J = 6.2 Hz, 6H).

[0250] Example 24: Ethyl 2-methyl-6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridine-3-carboxylate [ka] 50 mg of colorless solid. HPLC / MS m / z: 473.2 [M+H] + , Rt (N): 1.34 min. 1H NMR (400 MHz, DMSO-d6): δ 8.81-8.79 (m, 1H), 8.27 (s, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.01-7.95 (m, 1H), 7.92 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.94 (d, J = 5.8 Hz, 1H), 4.67 (s, 2H), 4.33 (q, J = 7.1 Hz, 2H), 3.90-3.85 (m, 2H), 3.85-3.80 (m, 2H), 2.81 (s, 3H), 2.67 (s, 3H), 1.33 (t, J = 7.1 Hz, 3H).

[0251] Example 25: 2-methyl-6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-propoxy-5H,6H,7H-pyrrolo[3,4-b]pyridin-5-one [ka]

[0252] Example 25.1: A solution of methyl 2-hydroxy-6-methylpyridine-3-carboxylate (4.28 g, 25.604 mmol) and NBS (4.56 g, 25.620 mmol) in DMF (100.0 mL) was stirred at room temperature under a nitrogen atmosphere for 4 h and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 7:1) to give 6.0 g (95%) of methyl 5-bromo-2-hydroxy-6-methylpyridine-3-carboxylate as a colorless solid. HPLC / MS m / z: 246.05 [M+H] + , Rt (B): 0.61 min.

[0253] Example 25.2: A solution of Intermediate 25.1 (5.96 g, 24.222 mmol), 1,1,1-trifluoro-N-phenyl-N-trifluoromethanesulfonylmethanesulfonamide (12.45 g, 34.849 mmol), and DIPEA (9.00 g, 69.684 mmol) in DMF (50.0 mL) was stirred overnight at room temperature under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 1:1) to give 6.69 g (73%) of methyl 5-bromo-6-methyl-2-(trifluoromethanesulfonyloxy)pyridine-3-carboxylate as a yellow oil. HPLC / MS m / z: 380.00 [M+H] + , Rt (B): 1.12 min.

[0254] Example 25.3: Intermediate 25.2 (620.8 mg, 1.642 mmol) and tributyl(ethenyl)stannane (542.5 mg, 1.711 mmol) were dissolved in DMF (10.0 mL). Pd(PPh3)2Cl2 (115.0 mg, 0.164 mmol) was added, and the reaction mixture was stirred at 90 °C under a nitrogen atmosphere for 16 h. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 3:2) to give 415.0 mg (99%) of methyl 5-bromo-2-ethenyl-6-methylpyridine-3-carboxylate as a brown oil. HPLC / MS m / z: 256.10 [M+H] + , Rt (B): 1.08 min.

[0255] Example 25.4: To a stirred solution of intermediate 25.3 (0.88 g, 3.467 mmol) and NaIO (1.48 g, 6.919 mmol) in THF (10.0 mL) and water (2.0 mL), potassium osmate(VI) dihydrate (0.13 g, 0.345 mmol) was added and the mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo and the crude product (1.2 mg) was used in the next step without further purification.

[0256] Example 25.5: A solution of the crude aldehyde (192.0 mg, 0.744 mmol) in DCE (5.0 mL) was treated with tert-butyl N-(2-aminoethyl)carbamate (119.2 mg, 0.744 mmol) and sodium triacetoxyborohydride (315.4 mg, 1.488 mmol) and stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure, and the residue was purified by RP flash chromatography to give 139 mg (51%) of N-(2-{3-bromo-2-methyl-5-oxo-5H,6H,7H-pyrrolo[3,4-b]pyridin-6-yl}ethyl)carbamate as a colorless solid. HPLC / MS m / z: 372.15 [M+H] + , Rt (B): 0.81 min.

[0257] Example 25.6: To a solution of intermediate 25.4 (308.0 mg, 0.832 mmol) and propan-1-ol (50.0 mg, 0.832 mmol) in toluene (5.0 mL) was added CsCO (542.2 mg, 1.664 mmol) and tBuXPhos Pd G (66.1 mg, 0.083 mmol), and the resulting mixture was stirred at 90 °C under a nitrogen atmosphere for 16 h. The reaction mixture was concentrated under reduced pressure, and the residue was purified by RP flash chromatography to afford 198 mg (68%) of tert-butyl N-(2-{2-methyl-5-oxo-3-propoxy-5H,6H,7H-pyrrolo[3,4-b]pyridin-6-yl}ethyl)carbamate as a yellow solid. HPLC / MS m / z: 350.30 [M+H] + , Rt (P): 1.03 min.

[0258] Example 25.7: Intermediate 25.5 (198.0 mg, 0.566 mmol) was dissolved in dioxane (2.0 mL) and treated with a solution of HCl in dioxane (4 M, 2.0 mL), and the mixture was stirred at room temperature for 4 h. The mixture was basified to pH 8 by adding aqueous NaHCO solution. The mixture was concentrated in vacuo, and the residue was purified by RP flash chromatography to give 120 mg (84%) of 6-(2-aminoethyl)-2-methyl-3-propoxy-5H,6H,7H-pyrrolo[3,4-b]pyridin-5-one as a colorless solid. HPLC / MS m / z: 250.25 [M+H] + , Rt (P): 0.69 min.

[0259] Example 25.8: Prepared as described for Compound 20.5. Yield: 28 mg (15%) of 2-methyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-propoxy-5H,6H,7H-pyrrolo[3,4-b]pyridin-5-one as a colorless solid. HPLC / MS m / z: 459.15 [M+H] + , Rt (D): 1.19 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.83 (s, 1H), 8.14 (d, J = 8.6 Hz, 1H), 8.01 (t, J = 5.6 Hz, 1H), 7.95 (d, J = 5.8 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.46 (s, 1H), 6.95 (d, J = 5.7 Hz, 1H), 4.50 (s, 2H), 4.03 (t, J = 6.4 Hz, 2H), 3.88-3.79 (m, 4H), 2.68 (s, 3H), 2.46 (s, 3H), 1.80-1.69 (m, 2H), 1.00 (t, J = 7.4 Hz, 3H).

[0260] Example 26: 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-6-propoxy-1H,2H,3H-pyrrolo[3,4-c]pyridin-1-one [ka]

[0261] Example 26.1: To a solution of methyl 5-bromo-2-oxo-1,2-dihydropyridine-4-carboxylate (3.80 g, 16.377 mmol) and 1-iodopropane (5.57 g, 32.754 mmol) in toluene (40.0 mL), AgCO (6.77 g, 24.566 mmol) was added, and the reaction mixture was stirred at 100 °C under a nitrogen atmosphere for 2 h. The mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether - 1:4) to afford 4.0 g (89%) of methyl 5-bromo-2-propoxypyridine-4-carboxylate as a colorless solid. HPLC / MS m / z: 276.1 [M+H] + , Rt (P): 1.28 min.

[0262] Example 26.2: Intermediate 26.1 (1.92 g, 7.004 mmol) was reacted with tributyl(ethenyl)stannane as described for compound 27.3. Yield: 1.16 g (75%) of methyl 5-ethenyl-2-propoxypyridine-4-carboxylate as a yellow liquid. HPLC / MS m / z: 222.05 [M+H] + , Rt (B): 1.05 min.

[0263] Example 26.3: To a solution of intermediate 26.2 (965.0 mg, 4.361 mmol) and NaIO (1.87 g, 8.743 mmol) in 1,4-dioxane (10.0 mL) and water (2.0 mL), KMnO (1.03 g, 6.518 mmol) was added, and the mixture was stirred at room temperature for 30 min. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether - 2:3) to give 507 mg (52%) of methyl 5-formyl-2-propoxypyridine-4-carboxylate as a yellow solid. HPLC / MS m / z: 224.05 [M+H] + , Rt (D): 1.21 min.

[0264] Example 26.4: To a solution of intermediate 26.3 (507.0 mg, 2.271 mmol) and benzyl N-(2-aminoethyl)carbamate (441.1 mg, 2.271 mmol) in dichloroethane (10.0 mL), sodium triacetoxyborohydride (721.6 mg, 3.405 mmol) was added, and the mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether - 2:3) to yield 300.0 mg (35%) of benzyl N-(2-{1-oxo-6-propoxy-1H,2H,3H-pyrrolo[3,4-c]pyridin-2-yl}ethyl)carbamate as a yellow solid. HPLC / MS m / z: 370.25 [M+H] + , Rt (P): 1.05 min.

[0265] Example 26.5: Intermediate 26.4 (277.0 mg, 0.750 mmol) was stirred in aqueous HCl solution (2N, 5.0 mL) at 50° C. for 2 h. The mixture was cooled to room temperature, neutralized to pH 7 with aqueous NaHCO solution, and concentrated in vacuo. The residue was purified by reverse flash chromatography to yield 158 mg (89%) of 2-(2-aminoethyl)-6-propoxy-1H,2H,3H-pyrrolo[3,4-c]pyridin-1-one as an off-white solid. HPLC / MS m / z: 236.25 [M+H] + , Rt (P): 0.62 min.

[0266] Example 26.6: Prepared as described for Compound 20.5. Yield: 23 mg (9%) of 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-6-propoxy-1H,2H,3H-pyrrolo[3,4-c]pyridin-1-one as an off-white solid. HPLC / MS m / z: 445.25 [M+H] + , Rt (D): 1.51 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.70 (d, J = 1.5 Hz, 1H), 8.29 (d, J = 1.0 Hz, 1H), 8.02 (dd, J = 8.5, 1.5 Hz, 1H), 7.92 (d, J = 5.6 Hz, 1H), 7.82 (d, J = 5.7 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 6.87-6.77 (m, 2H), 4.47 (s, 2H), 4.11 (t, J = 6.7 Hz, 2H), 3.76-3.64 (m, 4H), 2.56 (s, 3H), 1.69-1.51 (m, 2H), 0.83 (t, J = 7.4 Hz, 3H).

[0267] Example 27: Ethyl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate [ka]

[0268] Example 27.1: A solution of piperidine-2,4-dione (14.25 g, 125.978 mmol), 2,2-dimethoxyethan-1-amine (13.24 g, 125.978 mmol), and 4-methylbenzene-1-sulfonic acid (2.17 g, 12.601 mmol) in toluene (150.0 mL) was stirred at 110° C. overnight. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in dichloromethane (200 mL) and treated with TFA (143.74 g, 1260.612 mmol), and the resulting mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 9:1) to give 8.31 g (48%) of 1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridin-4-one as a yellow solid. HPLC / MS m / z: 137.20 [M+H] + , Rt (B): 0.22 min.

[0269] Example 27.2: A solution of 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (6.20 g, 21.684 mmol) and Intermediate 27.1 (8.31 g, 61.035 mmol) in DMF (100.0 mL) was stirred at −60° C. for 20 min under a nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol—9:1) to yield 11.79 g (90%) of 2-bromo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridin-4-one as a colorless solid. HPLC / MS m / z: 215.10 / 217.05 [M+H]+ , Rt (B): 0.47 min.

[0270] Example 27.3: Intermediate 27.2 (4.70 g, 21.856 mmol) was dissolved in DMF (60.0 mL) and treated with CsCO (14.44 g, 44.319 mmol) and methyl iodide (3.47 g, 24.447 mmol). The reaction mixture was stirred at room temperature overnight and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 13:1) to give 4.08 g (82%) of 2-bromo-1-methyl-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridin-4-one as a colorless solid. HPLC / MS m / z: 229.10 / 231.15 [M+H] + , Rt (B): 0.61 min.

[0271] Example 27.4: To a solution of intermediate 27.3 (1.70 g, 7.421 mmol) in ethanol (15.0 mL) in a pressure tank was added Pd(dppf)Cl.CHCl (0.68 g, 0.833 mmol) and triethylamine (2.53 g, 25.002 mmol). The mixture was purged with nitrogen for 5 min, then pressurized to 20 atm with carbon monoxide and stirred at 100 °C overnight. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 10:1) to afford 1.56 g (95%) of ethyl 1-methyl-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate as a brown semi-solid. HPLC / MS m / z: 223.20 [M+H] + , Rt (B): 0.62 min.

[0272] Example 27.5: Intermediate 27.4 (785.4 mg, 3.534 mmol) was dissolved in THF (10.0 mL) and sodium hydride (60%, 0.76 g, 19.000 mmol) was added at 0 °C. The mixture was warmed to room temperature and stirred for 16 h. 2-Bromoacetonitrile (2.85 g, 23.760 mmol) was added and the mixture was stirred at room temperature for 2 d. The reaction was quenched with EtOH, and the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 10:1) to produce 199 mg (22%) of ethyl 5-(cyanomethyl)-1-methyl-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate as a light brown solid. HPLC / MS m / z: 262.30 [M+H] + , Rt (B): 0.70 min.

[0273] Example 27.6: Intermediate 27.5 (184.0 mg, 0.704 mmol) was hydrogenated in EtOH (5.0 mL) in the presence of Raney Ni (10%, 200.0 mg, 0.233 mmol) at room temperature for 16 h. The reaction mixture was filtered, and the filter cake was washed with dichloromethane. The filtrate was concentrated under reduced pressure, and the residue was purified by RP flash chromatography to give 49 mg (26%) of ethyl 5-(2-aminoethyl)-1-methyl-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate as a brown solid. HPLC / MS m / z: 266.25 [M+H] + , Rt (B): 0.56 min.

[0274] Example 27.7: Prepared as described for Compound 20.5. Yield: 40 mg (46%) of ethyl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate as a colorless solid. HPLC / MS m / z: 475.00 [M+H] +, Rt (L): 0.63 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.86 (s, 1H), 8.22-8.08 (m, 1H), 8.03-7.91 (m, 2H), 7.82 (d, J = 8.6 Hz, 1H), 7.00 (s, 1H), 6.93 (d, J = 5.7 Hz, 1H), 4.19 (q, J = 7.1 Hz, 2H), 3.73 (s, 3H), 3.70-3.66 (m, 3H), 3.63 (t, J = 6.8 Hz, 3H), 2.87 (t, J = 6.8 Hz, 2H), 2.71-2.64 (m, 3H), 2.28-2.24 (m, 1H), 1.25 (t, J = 7.1 Hz, 3H).

[0275] The following compounds were prepared similarly: Example 28: Propan-2-yl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate [ka] 9 mg of colorless solid. HPLC / MS m / z: 489.05 [M+H] + , Rt (B): 0.72 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.97 (s, 1H), 8.19 (s, 1H), 7.92 (d, J = 5.8 Hz, 2H), 7.07 (s, 1H), 6.98 (s, 1H), 5.11-4.97 (m, 1H), 3.79-3.63 (m, 8H), 2.91 (t, J = 6.7 Hz, 2H), 2.77-2.67 (m, 4H), 1.27 (d, J = 6.2 Hz, 6H).

[0276] Example 29: Ethyl 1-ethyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate [ka] 11 mg of colorless solid. HPLC / MS m / z: 489.20 [M+H] + , Rt (O): 1.49 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.89 (s, 1H), 8.18-8.11 (m, 1H), 8.05-8.01 (m, 1H), 7.97 (d, J = 5.7 Hz, 1H), 7.85 (d, J = 8.5 Hz, 1H), 7.04 (s, 1H), 6.96 (d, J = 5.8 Hz, 1H), 4.31-4.17 (m, 3H), 3.90 (q, J = 7.1 Hz, 1H), 3.73- 3.57 (m, 6H), 2.96-2.81 (m, 2H), 2.70 (s, 3H), 1.28 (t, J = 7.1 Hz, 3H), 1.24-1.14 (m, 3H).

[0277] Example 30: Propan-2-yl 1-ethyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate [ka] 2mg. HPLC / MS m / z: 503.35 [M+H] + , Rt (B): 0.88 min. 1H NMR (300 MHz, DMSO-d6): δ 8.89 (s, 1H), 8.15 (dd, J = 8.5, 1.5 Hz, 1H), 8.06-7.93 (m, 2H), 7.85 (d, J = 8.5 Hz, 1H), 7.01 (s, 1H), 6.96 (d, J = 5.8 Hz, 1H), 5.12-4.98 (m, 1H), 4.25 (q, J = 7.0 Hz, 2H), 3.87-3.49 (m, 6H), 2.91 (t, J = 6.7 Hz, 2H), 2.70 (s, 3H), 1.27 (d, J = 6.2 Hz, 6H), 1.21 (t, J = 7.1 Hz, 3H).

[0278] Example 31: Ethyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-7-oxo-4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate [ka]

[0279] Example 31.1: 2-Bromo-4H,5H,6H,7H-thieno[2,3-c]pyridin-7-one (447.0 mg, 1.926 mmol) was dissolved in dry THF (15.0 mL), sodium hydride (60% dispersion in mineral oil; 231.1 mg, 5.779 mmol) was added, and the suspension was cooled in an ice bath under a nitrogen atmosphere and stirred for 45 min. Bromoacetonitrile (69.2 mg, 5.779 mmol) was added, and the reaction mixture was allowed to warm to room temperature and stirred for 2 h. The reaction was quenched with aqueous saturated NH4Cl solution and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by flash chromatography to yield 415 mg (79%) of 2-{2-bromo-7-oxo-4H,5H,6H,7H-thieno[2,3-c]pyridin-6-yl}acetonitrile as colorless crystals. HPLC / MS m / z: 270.7 / 272.8 [M+H] +, Rt (G): 1.40 min.

[0280] Example 31.2: Intermediate 31.1 (229.3 mg, 0.846 mmol) was dissolved in dry methanol (3.5 mL) and dry THF (11 mL). Triethylamine (175.9 μl, 1.269 mmol), Pd(dppf)Cl2.CHCl2 (41.3 mg, 0.052 mmol), and 1,1-bis-(diphenylphosphino)-ferrocene (37.5 mg, 0.068 mmol) were added under a nitrogen atmosphere. The reactor was pressurized to 4.0 bar with carbon monoxide, and the reaction mixture was stirred at 100 °C overnight. The reaction mixture was filtered, the filtrate evaporated to dryness, and the residue purified by flash chromatography to give 194 mg (92%) of methyl 6-(cyanomethyl)-7-oxo-4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate as a red oil. HPLC / MS m / z: 250.9 [M+H] + , Rt (G): 1.26 min.

[0281] Example 31.3: Hydrogenation of intermediate 31.2 (194.0 mg, 0.775 mmol) was carried out over Raney Ni. Yield: 88 mg (39%) of methyl 6-(2-aminoethyl)-7-oxo-4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate hydrochloride as a colorless solid. HPLC / MS m / z: 254.9 [M+H] + , Rt (G): 0.97 min.

[0282] Example 31.4: Prepared as described for Compound 20.5. Yield: 20 mg (22%) of methyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-7-oxo-4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate as a colorless oil. HPLC / MS m / z: 463.9 [M+H] + , Rt (M): 1.73 min.

[0283] Example 31.5: Transesterification of intermediate 31.4 (12.0 mg, 0.026 mmol) was carried out as described in Example 2. Yield: 9.5 mg (77%) of ethyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-7-oxo-4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate as a colorless oil. HPLC / MS m / z: 477.8 [M+H] + , Rt (M): 1.80 min. 1 H NMR (700 MHz, DMSO-d6): δ 13.47-12.58 (m, 1H), 9.05-8.92 (m, 1H), 8.35-8.21 (m, 1H), 8.05-7.89 (m, 1H), 7.92-7.86 (m, 1H), 7.69 (s, 1H), 7.21-7.04 (m, 1H), 4.30 (q, J = 7.1 Hz, 2H), 3.82-3.75 (m, 4H), 3.71 (t, J = 7.0 Hz, 2H), 2.90 (t, J = 7.0 Hz, 2H), 2.70 (s, 3H), 1.29 (t, J = 7.1 Hz, 3H).

[0284] Example 32: Ethyl 5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4H,5H,6H-thieno[2,3-c]pyrrole-2-carboxylate [ka]

[0285] Example 32.1: Methyl 4,5-bis(chloromethyl)thiophene-2-carboxylate (47.5 mg, 0.199 mmol) was dissolved in dry ethanol (0.5 mL). Anhydrous sodium carbonate (56.8 mg, 0.536 mmol) and N-(2-aminoethyl)carbamic acid tert-butyl ester (35.0 mg, 0.219 mmol) were added, and the mixture was stirred at 100° C. for 1 h. The reaction was filtered and washed with ethanol. The filtrate was evaporated to dryness, and the residue was purified by flash chromatography to give 38 mg (59%) of methyl 5-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4H,5H,6H-thieno[2,3-c]pyrrole-2-carboxylate as a colorless solid. HPLC / MS m / z: 326.9 [M+H] + , Rt (H): 0.66 min.

[0286] Example 32.2: Intermediate 32.1 (585.0 mg, 1.792 mmol) was suspended in 1,4-dioxane (6.0 mL). A solution of HCl in dioxane (4N, 2.69 mL) was added and the suspension was stirred at room temperature overnight. The reaction mixture was evaporated to dryness to yield 535 mg (100%) of methyl 5-(2-aminoethyl)-4H,5H,6H-thieno[2,3-c]pyrrole-2-carboxylate dihydrochloride as a gray-green solid. HPLC / MS m / z: 226.9 [M+H] + , Rt (H): 0.19 min.

[0287] Example 32.3: Under a nitrogen atmosphere, 1-chloro-7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinoline (133.0 mg, 0.541 mmol), Intermediate 32.2 (194.4 mg, 0.650 mmol), (R)-(+)-2,2'-bis(diphenylphosphino)-1,1'-binaphthalene (20.2 mg, 0.032 mmol), and palladium(II) acetate (47% Pd; 7.3 mg, 0.032 mmol) were suspended in dry toluene (0.6 mL). Potassium tert-butylate (182.3 mg, 1.624 mmol) was added, and the reaction mixture was heated to 85 °C and stirred overnight. The mixture was cooled to room temperature, filtered through Celite, and washed with dichloromethane / methanol. The filtrate was diluted with demineralized water and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to give 54 mg (23%) of methyl 5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4H,5H,6H-thieno[2,3-c]pyrrole-2-carboxylate as a colorless solid. HPLC / MS m / z: 435.8 [M+H] + , Rt (H): 0.69 min. 1 H NMR (500 MHz, DMSO-d6): δ 8.90-8.88 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 7.97 (d, J = 5.7 Hz, 1H), 7.90-7.86 (m, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.61 (s, 1H), 6.94 (d, J = 5.4 Hz, 1H), 4.12-4.04 (m, 2H), 3.94-3.87 (m, 2H), 3.80 (s, 3H), 3.69 (q, J = 6.3 Hz, 2H), 3.10-3.03 (m, 2H), 2.70 (s, 3H).

[0288] Example 32.4. Transesterification of intermediate 32.3 (18.0 mg, 0.041 mmol) was carried out as described in Example 2. Yield: 3 mg (16%) of ethyl 5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4H,5H,6H-thieno[2,3-c]pyrrole-2-carboxylate. HPLC / MS m / z: 449.8 [M+H] + , Rt (M): 1.72 min.

[0289] Example 33: Propan-2-yl 5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4H,5H,6H-thieno[2,3-c]pyrrole-2-carboxylate [ka] Saponification of intermediate 32.3 (48.0 mg, 0.110) and esterification of the acid were carried out as described in Example 3. Yield: 9 mg (21%) of propan-2-yl 5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4H,5H,6H-thieno[2,3-c]pyrrole-2-carboxylate as a colorless solid. HPLC / MS m / z: 463.8 [M+H] + , Rt (H): 0.73 min. 1H NMR (400 MHz, DMSO-d6): δ 8.90-8.88 (m, 1H), 8.15 (dd, J = 8.5, 1.6 Hz, 1H), 7.97 (d, J = 5.7 Hz, 1H), 7.92-7.87 (m, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.57 (s, 1H), 6.95 (d, J = 5.8 Hz, 1H), 5.12-5.02 (m, 1H), 4.17-4.06 (m, 2H), 4.00-3.88 (m, 2H), 3.70 (q, J = 6.2 Hz, 2H), 3.16-3.05 (m, 2H), 2.70 (s, 3H), 1.28 (d, J = 6.2 Hz, 6H).

[0290] Example 34: Ethyl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-1H,4H,5H,6H-pyrrolo[2,3-c]pyrrole-2-carboxylate [ka]

[0291] Example 34.1: To a stirred solution of POCl3 (50.1 g, 327.760 mmol) in dichloroethane (300 mL) was added DMF (23.9 g, 347.760 mmol) very slowly at 0 °C. The resulting mixture was allowed to warm to room temperature and stirred for 1 h. Ethyl 4-bromo-1H-pyrrole-2-carboxylate (15.00 g, 65.352 mmol, 1.00 equiv, 95%) was added at room temperature, and the resulting mixture was heated to 80 °C and stirred for 1 h. The reaction mixture was cooled to room temperature and concentrated under vacuum. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 3:1) to yield 14.6 g (91%) of ethyl 4-bromo-5-formyl-1H-pyrrole-2-carboxylate as a yellow solid. HPLC / MS m / z: 245.9 [M+H] + , Rt (A): 0.78 min.

[0292] Example 34.2: To a stirred solution of intermediate 34.1 (14.85 g, 56.287 mmol) in toluene (160.0 mL) was added tetramethylammonium fluoride (15.72 g, 168.796 mmol) at room temperature. The resulting mixture was heated to 100° C. and stirred for 16 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate—92:8) to afford 13 g (88%) of ethyl 4-bromo-5-formyl-1-methylpyrrole-2-carboxylate as a colorless solid. HPLC / MS m / z: 262.0 [M+H] + , Rt (J): 0.78 min.

[0293] Example 34.3: To a stirred solution of Intermediate 34.2 (9.93 g, 38.180 mmol) and (tributylstannyl)methanol (17.17 g, 53.473 mmol) in toluene (115.0 mL) was added Pd(PPh3)4 (0.44 g, 0.381 mmol) at room temperature under a nitrogen atmosphere. The resulting mixture was heated to 110 °C and stirred for 4 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 6:4) to afford 7.9 g (98%) of ethyl 5-formyl-4-(hydroxymethyl)-1-methylpyrrole-2-carboxylate as a yellow solid. HPLC / MS m / z: 212.1 [M+H] + , Rt (J): 0.57 min.

[0294] Example 34.4: Intermediate B3 (266.6 mg, 0.990 mmol) and Intermediate 34.3 (250.8 mg, 1.187 mmol) were dissolved in dichloroethane (5.0 mL), a trace of acetic acid was added, and the mixture was stirred at 50 °C for 16 h. The reaction mixture was cooled to room temperature, sodium borohydride (187.2 mg, 4.948 mmol) was added, and the mixture was stirred at room temperature for 1 h. The reaction was quenched with aqueous saturated NH4Cl solution (10 mL), and the resulting mixture was extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 9 / 1) to give 106 mg (23%) of 4-(hydroxymethyl)-1-methyl-5-{[(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)amino]methyl}-1H-pyrrole-2-carboxylate as a colorless solid. HPLC / MS m / z: 465.25 [M+H] + , Rt (J): 0.58 min.

[0295] Example 34.5: To a stirred solution of Intermediate 34.4 (106.0 mg, 0.228 mmol) in dichloromethane (2.0 mL) was added thionyl chloride (56.0 mg, 0.471 mmol) at room temperature, and the resulting mixture was stirred at room temperature for 1 h. The reaction mixture was concentrated in vacuo to afford 109 mg (99%) of ethyl 4-(chloromethyl)-1-methyl-5-{[(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)amino]methyl}-1H-pyrrole-2-carboxylate as an off-white solid. HPLC / MS m / z: 479.25 [M+H] + , Rt (B): 0.74 min.

[0296] Example 34.6: A solution of intermediate 34.5 (95.0 mg, 0.196 mmol) in DMF (2.0 mL) was treated with potassium carbonate (94.0 mg, 0.682 mmol), and the mixture was heated to 80° C. and stirred for 16 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The crude product was purified by preparative HPLC to yield 3 mg (3%) of ethyl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-1H,4H,5H,6H-pyrrolo[2,3-c]pyrrole-2-carboxylate as a brown oil. HPLC / MS m / z: 447.10 [M+H] + , Rt (B): 0.62 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.89 (s, 1H), 8.18-8.11 (m, 1H), 7.97 (d, J = 5.7 Hz, 1H), 7.90-7.80 (m, 2H), 6.94 (d, J = 5.7 Hz, 1H), 6.62 (s, 1H), 4.17 (q, J = 7.1 Hz, 2H), 3.89 (s, 2H), 3.75 (d, J = 6.8 Hz, 4H), 3.67 (q, J = 6.4 Hz, 2H), 3.03 (t, J = 6.8 Hz, 2H), 2.70 (s, 3H), 1.24 (t, J = 7.2 Hz, 4H).

[0297] Example 35: Ethyl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H-pyrrolo[2,3-c]pyrrole-2-carboxylate [ka]

[0298] Example 35.1: To a stirred solution of intermediate 34.2 (803.0 mg, 3.087 mmol) and formyl acetate (2.91 g, 33.045 mmol) in DMF (1.0 mL) was added Pd(dppf)Cl2.CHCl2 (1.52 g, 1.861 mmol) at room temperature under a nitrogen atmosphere, followed by a trace of triethylamine. The resulting mixture was stirred at 90 °C for 3 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by RP flash chromatography to give 530 mg (76%) of 5-(ethoxycarbonyl)-2-formyl-1-methyl-1H-pyrrole-3-carboxylic acid as a yellow solid. HPLC / MS m / z: 226.15 [M+H] + , Rt (J): 0.65 min.

[0299] Example 35.2: Intermediate 35.1 (488.9 mg; 2.171 mmol) was converted as described for compound 34.4. After purification by silica gel column chromatography (eluent: dichloromethane / methanol - 9:1), 832 mg (80%) of 5-(ethoxycarbonyl)-1-methyl-2-{[(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)amino]methyl}-1H-pyrrole-3-carboxylic acid was isolated as a brown solid. HPLC / MS m / z: 479.25 [M+H] + , Rt (J): 0.63 min.

[0300] Example 35.3: To a stirred solution of intermediate 35.2 (677.6 mg, 1.416 mmol) in DMF (6.0 mL) was added 1-methyl-1H-imidazole (348.7 mg, 4.247 mmol) at room temperature, and the mixture was stirred at room temperature for 15 min. [Chloro(dimethylamino)methylidene]dimethylazanium; Hexafluoro-λ 5HCl phosphanide (476.9 mg, 1.700 mmol) was added at room temperature, and the mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol - 9:1) to give 584 mg (90%) of ethyl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H-pyrrolo[2,3-c]pyrrole-2-carboxylate as a colorless solid. HPLC / MS m / z: 461.25 [M+H] + , Rt (J): 0.71 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.73 (s, 1H), 8.02 (d, J = 8.6 Hz, 1H), 7.92-7.80 (m, 2H), 7.72 (d, J = 8.5 Hz, 1H), 6.83 (d, J = 5.8 Hz, 1H), 6.76 (d, J = 1.1 Hz, 1H), 4.40 (s, 2H), 4.11 (q, J = 7.1 Hz, 2H), 3.71 (s, 3H), 3.61 (s, 4H), 2.57 (s, 3H), 1.21-1.10 (m, 3H).

[0301] The following examples were prepared in a similar manner: Example 36: Propan-2-yl 1-methyl-5-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4-oxo-1H,4H,5H,6H-pyrrolo[2,3-c]pyrrole-2-carboxylate [ka] Saponification of compound 35 and esterification of the corresponding acid using standard conditions provided compound 36. 25 mg colorless solid. HPLC / MS m / z: 475.30 [M+H] + , Rt (Q): 2.06 min. 1H NMR (300 MHz, DMSO-d6): δ 8.73 (s, 1H), 8.07-7.98 (m, 1H), 7.92-7.80 (m, 2H), 7.72 (d, J = 8.5 Hz, 1H), 6.83 (d, J = 5.7 Hz, 1H), 6.73 (s, 1H), 5.01-4.84 (m, 1H), 4.39 (s, 2H), 3.70 (s, 3H), 3.60 (d, J = 2.5 Hz, 4H), 2.57 (s, 3H), 1.16 (d, J = 6.2 Hz, 6H).

[0302] Example 37: 2-{2-[7-(5-methyl-[1,2,4]oxadiazol-3-yl)-isoquinolin-1-ylamino]-ethyl}-1-oxo-1,2-dihydro-isoquinoline-7-carboxylic acid ethyl ester [ka]

[0303] Example 37.1: A solution of 7-bromoisoquinolin-1-ol (1.00 g, 4.463 mmol), tert-butyl N-(2-chloroethyl)carbamate (963 mg, 5.356 mmol), and potassium carbonate (1.90 g, 13.748 mmol) in DMF (11.0 mL) was heated to 80° C. and stirred for 12 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by PP flash chromatography to give 1.48 g (90%) of tert-butyl N-[2-(7-bromo-1-oxoisoquinolin-2-yl)ethyl]carbamate as a colorless solid. HPLC / MS m / z: 367.15 [M+H] + , Rt (B): 0.97 min.

[0304] Example 37.2: To a solution of intermediate 37.1 (494.5 mg, 1.342 mmol) in ethanol (5.0 mL) in a pressure tank was added Pd(dppf)Cl2.CHCl2 (100.8 mg, 0.123 mmol) and triethylamine (373.0 mg, 3.686 mmol). The mixture was purged with nitrogen for 5 min, pressurized to 20 atm with carbon monoxide, and stirred at 100 °C overnight. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids. The filtrate was evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 1:4) to yield 475 mg (98%) of ethyl 2-[2-[(tert-butoxycarbonyl)amino]ethyl]-1-oxoisoquinoline-7-carboxylate as a brown solid. HPLC / MS m / z: 361.25 [M+H] + , Rt (B): 0.94 min.

[0305] Example 37.3: Deprotection of intermediate 37.2 (475 mg, 1.318 mmol) with HCl in 1,4-dioxane was carried out as described for compound 25.7. Yield: 340 mg (99%) of ethyl 2-(2-aminoethyl)-1-oxo-1,2-dihydroisoquinoline-7-carboxylate as a brown solid. HPLC / MS m / z: 261.15 [M+H] + , Rt (B): 0.60 min.

[0306] Example 37.4: A solution of intermediate 37.3 (100 mg, 0.384 mmol), 1-chloro-7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinoline (18.9 mg, 0.077 mmol), and potassium carbonate (25.2 mg, 0.182 mmol) in dimethyl sulfoxide (2.6 mL) was stirred at 120° C. for 2 d. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by preparative HPLC to yield 3.5 mg (10%) of ethyl 2-(2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino]ethyl)-1-oxoisoquinoline-7-carboxylate as a colorless solid. HPLC / MS m / z: 470.05 [M+H] + , Rt (K): 1.07 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.85-8.75 (m, 2H), 8.20-8.08 (m, 2H), 8.02-7.96 (m, 1H), 7.92-7.79 (m, 2H), 7.70 (d, J = 8.4 Hz, 1H), 7.35 (d, J = 7.3 Hz, 1H), 6.93 (d, J = 5.8 Hz, 1H), 6.55 (d, J = 7.4 Hz, 1H), 4.42-4.24 (m, 4H), 3.89-3.82 (m, 2H), 2.65 (s, 3H), 1.35 (t, J = 7.1 Hz, 3H).

[0307] The following examples were prepared in a similar manner. Example 38: Ethyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6-dihydro-1,6-naphthyridine-3-carboxylate [ka] 25 mg off-white solid. HPLC / MS m / z: 471.10 [M+H] + , Rt (R): 0.64 min. 1H NMR (300 MHz, DMSO-d6): δ 9.29 (d, J = 2.2 Hz, 1H), 8.93 (d, J = 2.3 Hz, 1H), 8.76 (s, 1H), 8.12 (d, J = 8.5 Hz, 1H), 7.96 (s, 1H), 7.82 (d, J = 7.1 Hz, 2H), 7.62 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 5.8 Hz, 1H), 6.61 (d, J = 7.5 Hz, 1H), 4.49-4.35 (m, 2H), 4.30 (s, 2H), 3.87 (s, 2H), 2.65 (s, 3H), 1.36 (t, J = 7.1 Hz, 3H).

[0308] Example 39: Propan-2-yl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6-dihydro-1,6-naphthyridine-3-carboxylate [ka] 9 mg of colorless solid. HPLC / MS m / z: 485.20 [M+H] + , Rt (B): 0.86 min. 1 H NMR (300 MHz, DMSO-d6): δ 9.28 (d, J = 2.2 Hz, 1H), 8.91 (d, J = 2.2 Hz, 1H), 8.75 (s, 1H), 8.15-8.10(m,1H), 7.97 (s, 1H), 7.84-7.79 (m, 2H), 7.62 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 5.8 Hz, 1H), 6.61 (d, J = 7.5 Hz, 1H), 5.25-5.14 (m, J = 6.2 Hz, 1H), 4.29 (t, J = 5.7 Hz, 2H), 3.87 (d, J = 5.4 Hz, 2H), 2.65 (s, 3H), 1.35 (d, J = 6.3 Hz, 6H).

[0309] Example 40: tert-Butyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6-dihydro-1,6-naphthyridine-3-carboxylate [ka] 25 mg of yellow solid. HPLC / MS m / z: 499.15 [M+H] + , Rt (B): 0.91 min. 1 H NMR (300 MHz, DMSO-d6+D2O): δ 9.22 (d, J = 2.2 Hz, 1H), 8.84 (d, J = 2.3 Hz, 1H), 8.73 (d, J = 1.5 Hz, 1H), 8.14-8.08 (m, 1H), 7.84-7.76 (m, 2H), 7.61 (d, J = 7.6 Hz, 1H), 6.90 (d, J = 5.8 Hz, 1H), 6.59 (d, J = 7.6 Hz, 1H), 4.28 (t, J = 5.6 Hz, 2H), 3.85 (t, J = 5.6 Hz, 2H), 2.63 (s, 3H), 1.56 (s, 9H).

[0310] Example 41: Ethyl 6-(2-{[3-(5-methyl-1,2,4-oxadiazol-3-yl)phenyl]formamido}ethyl)-5-oxo-5,6-dihydro-1,6-naphthyridine-3-carboxylate [ka] 10 mg of colorless solid. HPLC / MS m / z: 448.15 [M+H] + , Rt (B): 0.88 min. 1H NMR (300 MHz, DMSO-d6): δ 9.31 (d, J = 2.2 Hz, 1H), 8.93 (d, J = 2.2 Hz, 1H), 8.80 (t, J = 5.8 Hz, 1H), 8.34 (t, J = 1.7 Hz, 1H), 8.13-8.07 (m, 1H), 7.95-7.89 (m, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.62 (t, J = 7.8 Hz, 1H), 6.71 (d, J = 7.5 Hz, 1H), 4.42-4.33 (m, 2H), 4.19 (t, J = 5.6 Hz, 2H), 3.66 (d, J = 5.8 Hz, 2H), 2.65 (s, 3H), 1.35 (t, J = 7.1 Hz, 3H).

[0311] Example 42: Ethyl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-3-carboxylate [ka] 53mg colorless solid. HPLC / MS m / z: 473.20 [M+H] + , Rt (B): 0.77 min. 1H NMR (400 MHz, DMSO-d6): δ 9.07 (d, J = 2.2 Hz, 1H), 8.84 (s, 1H), 8.51 (d, J = 2.2 Hz, 1H), 8.18-8.11 (m, 1H), 8.02 (t, J = 5.3 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.95 (d, J = 5.8 Hz, 1H), 4.40-4.30 (m, 2H), 3.87-3.76 (m, 4H), 3.73 (t, J = 6.7 Hz, 2H), 3.17 (t, J = 6.7 Hz, 2H), 2.68 (s, 3H), 1.33 (t, J = 7.1 Hz, 3H).

[0312] Example 43: Propan-2-yl 6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-3-carboxylate [ka] 38mg colorless solid. HPLC / MS m / z: 487.15 [M+H] + , Rt (B): 0.82 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.92 (d, J = 2.2 Hz, 1H), 8.71 (d, J = 1.5 Hz, 1H), 8.36 (d, J = 2.2 Hz, 1H), 8.04-7.99 (m, 1H), 7.91 (d, J = 5.6 Hz, 1H), 7.82 (d, J = 5.7 Hz, 1H), 7.70 (d, J = 8.5 Hz, 1H), 6.81 (d, J = 5.7 Hz, 1H), 5.13-4.95 (m, 1H), 3.60 (t, J = 6.7 Hz, 6H), 3.04 (t, J = 6.6 Hz, 2H), 2.55 (s, 3H), 1.20 (d, J = 6.2 Hz, 6H).

[0313] Example 44: Ethyl 2-methyl-6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-3-carboxylate [ka]

[0314] Example 44.1: Ethyl 5-cyano-2-hydroxy-6-methylpyridine-3-carboxylate (10.0 g, 48.497 mmol) was stirred in POCl (22.3 g, 145.436 mmol) at 90 °C for 16 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and quenched by the addition of ice water (20 mL). The resulting mixture was extracted with ethyl acetate, and the combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether - 2:3) to afford 7.71 g (71%) of ethyl 2-chloro-5-cyano-6-methylpyridine-3-carboxylate as a yellow solid. HPLC / MS m / z: 225.0 [M+H] + , Rt (B): 0.87 min.

[0315] Example 44.2: To a solution of Intermediate 44.1 (7.71 g, 34.316 mmol) and tributyl(ethenyl)stannane (13.06 g, 41.186 mmol) in DMF (80.0 mL) was added Pd(PPh3)2Cl2 (2.41 g, 3.434 mmol). The reaction mixture was heated to 100 °C under a nitrogen atmosphere and stirred for 16 h. The reaction mixture was cooled to room temperature, diluted with water (80 mL), and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether - 1:2) to afford 6.11 g (82%) of ethyl 5-cyano-2-ethenyl-6-methylpyridine-3-carboxylate as a colorless solid. HPLC / MS m / z: 217.25 [M+H] + , Rt (B): 0.98 min.

[0316] Example 44.3: A solution of Intermediate 44.2 (5.92 g, 27.390 mmol), benzyl N-(2-aminoethyl)carbamate (5.32 g, 27.390 mmol), and acetonitrile (15.0 mL) in methanol (15.0 mL) was treated with diethylamine (7.08 g, 54.799 mmol) and microwaved at 150 °C for 1 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether - 1:1) and by RP flash chromatography to afford 2 g (20%) of benzyl N-[2-(3-cyano-2-methyl-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridin-6-yl)ethyl]carbamate as a brown solid. HPLC / MS m / z: 365.20 [M+H] + , Rt (B): 0.85 min.

[0317] Example 44.4: Intermediate 44.3 (993.0 g, 2.724 mmol) was stirred in HCl (5.0 mL) at 100° C. for 16 h. The reaction mixture was cooled to room temperature, neutralized to pH 7 with aqueous NaHCO solution, and concentrated in vacuo. The residue was purified by RP flash chromatography to give 663 mg (98%) of 6-(2-aminoethyl)-2-methyl-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-3-carboxylic acid as a yellow solid. HPLC / MS m / z: 250.20 [M+H] + , Rt (B): 0.13 min.

[0318] Example 44.5: Intermediate 44.4 (351.5 mg, 1.410 mmol) was dissolved in ethanol (5.0 mL), treated with HSO (0.02 mL, 0.28 mmol), and stirred at 80 °C for 16 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by RP flash chromatography to afford 206 mg (53%) of ethyl 6-(2-aminoethyl)-2-methyl-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-3-carboxylate as a yellow solid. HPLC / MS m / z: 278.10 [M+H] + , Rt (B): 0.53 min.

[0319] Example 44.6: Prepared as described for Compound 20.5. Yield: 28 mg (9%) of ethyl 2-methyl-6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-3-carboxylate as an off-white solid. HPLC / MS m / z: 487.20 [M+H] + , Rt (B): 0.83 min. 1H NMR (300 MHz, DMSO-d6): δ 8.85 (s, 1H), 8.45 (s, 1H), 8.15 (dd, J = 8.5, 1.5 Hz, 1H), 8.04 (d, J = 5.2 Hz, 1H), 7.96 (d, J = 5.7 Hz, 1H), 7.85 (d, J = 8.6 Hz, 1H), 6.96 (d, J = 5.8 Hz, 1H), 4.38-4.25 (m, 2H), 3.84-3.75 (m, 4H), 3.71 (t, J = 6.7 Hz, 2H), 3.10 (t, J = 6.6 Hz, 2H), 2.72 (d, J = 17.5 Hz, 6H), 1.32 (t, J = 7.1 Hz, 3H).

[0320] The following examples were obtained in a similar manner. Example 45: Propan-2-yl 2-methyl-6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-5-oxo-5,6,7,8-tetrahydro-1,6-naphthyridine-3-carboxylate [ka] 68 mg of colorless solid. HPLC / MS m / z: 501.30 [M+H] + , Rt (S): 1.58 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.85 (s, 1H), 8.42 (s, 1H), 8.15 (dd, J = 8.5, 1.5 Hz, 1H), 7.92-8.05 (m, 2H), 7.85 (d, J = 8.5 Hz, 1H), 6.96 (d, J = 5.8 Hz, 1H), 5.05-5.22 (m, 1H), 3.77-3.84 (m, 3H), 3.70 (t, J = 6.6 Hz, 3H), 3.09 (t, J = 6.6 Hz, 2H), 2.72 (d, J = 15.9 Hz, 6H), 1.32 (d, J = 6.3 Hz, 6H).

[0321] Example 46: Methyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2H,3H-imidazo[1,5-a]pyrazine-6-carboxylate [ka]

[0322] Example 46.1: To a suspension of methyl 5-formylpyrazine-2-carboxylate (27.7 mg, 0.173 mmol) and Intermediate B3 (56.0 mg, 0.208 mmol) in dry dichloromethane (2.2 mL) was added sodium triacetoxyborohydride (55.1 mg, 0.260 mmol), and the mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give a pale red residue, which was purified by RP flash chromatography to yield 41 mg (56%) of methyl 5-{[(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)amino]methyl}pyrazine-2-carboxylate as a yellow solid. HPLC / MS m / z: 419.8 [M+H] + , Rt (H): 0.62 min. 1 H NMR (700 MHz, DMSO-d6): δ 9.06 (d, J = 1.4 Hz, 1H), 8.88-8.87 (m, 1H), 8.85 (d, J = 1.4 Hz, 1H), 8.14 (dd, J = 8.4, 1.5 Hz, 1H), 7.93 (d, J = 5.7 Hz, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.81 (t, J = 5.4 Hz, 1H), 6.92 (d, J = 5.6 Hz, 1H), 4.02 (s, 2H), 3.90 (s, 3H), 3.62 (q, J = 6.2 Hz, 2H), 2.88 (t, J = 6.4 Hz, 2H), 2.70 (s, 3H), 2.75-2.62 (m, 1H).

[0323] Example 46.2: Intermediate 46.1 (41.0 mg, 0.098 mmol) was suspended in dichloromethane (0.25 mL) and N-ethyldiisopropylamine (42.9 μL, 0.253 mmol), and the mixture was cooled to 0° C. in an ice bath. Bis(trichloromethyl)carbonate (29.1 mg, 0.098 mmol) dissolved in dichloromethane (0.49 mL) was added slowly. A clear solution was formed, which was allowed to warm to room temperature and stirred overnight. The reaction was evaporated to dryness and the residue was purified by flash chromatography to yield 23 mg (53%) of methyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2H,3H-imidazo[1,5-a]pyrazine-6-carboxylate as a yellow solid. HPLC / MS m / z: 445.8 [M+H] + , Rt (M): 1.61 min. 1 H NMR (700 MHz, DMSO-d6): δ 9.14-9.13 (m, 1H), 9.09-9.09 (m, 1H), 8.54 (dd, J = 8.4, 1.5 Hz, 1H), 8.30-8.29 (m, 1H), 8.26-8.25 (m, 1H), 8.12 (d, J = 8.4 Hz, 1H), 7.81 (d, J = 6.9 Hz, 1H), 7.37 (d, J = 6.9 Hz, 1H), 4.43-4.40 (m, 2H), 4.12-4.09 (m, 2H), 3.90 (s, 3H), 2.69 (s, 3H).

[0324] Example 47: Ethyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2H,3H-imidazo[1,5-a]pyrazine-6-carboxylate [ka]

[0325] Example 47.1: Example 46 (265.5 mg, 0.596 mmol) was dissolved in dry THF (14.8 mL) and water (7.4 mL) and treated with lithium hydroxide (36.0 mg, 1.490 mmol), and the reaction mixture was stirred at room temperature overnight. The reaction was diluted with water (40 mL) and acidified with HCl solution to pH 3-4. A precipitate formed, which was suction filtered, rinsed with water, and dried under high vacuum to give 145 mg (56%) of 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2H,3H-imidazo[1,5-a]pyrazine-6-carboxylic acid as a yellow solid. HPLC / MS m / z: 431.8 [M+H] + , Rt (M): 1.57 min.

[0326] Example 47.2: Intermediate 47.1 (48.5 mg, 0.112 mmol), 4-(dimethylamino)-pyridine (5.5 mg, 0.045 mmol), and DCC (37.1 mg, 0.180 mmol) were suspended in dry THF (0.3 mL) and DMF (0.9 mL). Ethanol (65.6 μl, 1.124 mmol) was added, and the reaction was stirred at room temperature overnight. The reaction was concentrated under reduced pressure, and the residue was purified by RP flash chromatography to yield 22 mg (43%) of ethyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2H,3H-imidazo[1,5-a]pyrazine-6-carboxylate as a yellow solid. HPLC / MS m / z: 459.8 [M+H] + , Rt (G): 1.30 min. 1H NMR (700 MHz, DMSO-d6): δ 8.80-8.79 (m, 1H), 8.46 (d, J = 1.7 Hz, 1H), 8.15 (dd, J = 8.4, 1.5 Hz, 1H), 7.99-7.98 (m, 1H), 7.96 (t, J = 5.7 Hz, 1H), 7.93 (d, J = 5.7 Hz, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.33 (d, J = 0.9 Hz, 1H), 6.97 (d, J = 5.8 Hz, 1H), 4.25 (q, J = 7.1 Hz, 2H), 4.15-4.12 (m, 2H), 3.84 (q, J = 5.6 Hz, 2H), 2.68 (s, 3H), 1.28 (t, J = 7.1 Hz, 3H).

[0327] Example 48: Propan-2-yl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2H,3H-imidazo[1,5-a]pyrazine-6-carboxylate [ka] Prepared as described in Example 47. Yield: 17 mg (32%) of a yellow solid. HPLC / MS m / z: 473.9 [M+H] + , Rt (H): 0.71 min. 1H NMR (700 MHz, DMSO-d6): δ 8.80-8.79 (m, 1H), 8.46 (d, J = 1.6 Hz, 1H), 8.15 (dd, J = 8.6, 1.5 Hz, 1H), 7.97-7.94 (m, 2H), 7.93 (d, J = 5.6 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.33 (s, 1H), 6.97 (d, J = 5.7 Hz, 1H), 5.11-5.05 (m, 1H), 4.14 (t, J = 5.7 Hz, 2H), 3.84 (q, J = 5.7 Hz, 2H), 2.69 (s, 3H), 1.28 (d, J = 6.2 Hz, 6H).

[0328] Example 49: Ethyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindole-5-carboxylate [ka]

[0329] Example 49.1: To a solution of methyl 2,3-dihydro-1H-isoindole-5-carboxylate hydrochloride (2.85 g, 13.339 mmol) and tert-butyl N-(2-chloroethyl)carbamate (4.09 g, 22.767 mmol) in DMF (40.0 mL), triethylamine (3.99 g, 39.431 mmol) was added, and the resulting mixture was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate - 1:1.5) to afford 1.95 g (46%) of methyl 2-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-2,3-dihydro-1H-isoindole-5-carboxylate as a brown oil. HPLC / MS m / z: 321.20 [M+H] + , Rt (J): 0.54 min.

[0330] Example 49.2: A solution of intermediate 49.1 (1.95 g, 6.086 mmol) in HCl / MeOH (20.0 mL) was stirred at room temperature for 2 h. The mixture was neutralized to pH 7 with aqueous saturated NaHCO3 solution (20 mL) and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure to give 1.28 g (86%) of methyl 2-(2-aminoethyl)-2,3-dihydro-1H-isoindole-5-carboxylate as a yellow solid. HPLC / MS m / z: 221.10 [M+H] + , Rt (J): 0.29 min.

[0331] Example 49.3: Prepared as described for Compound 20.5. Yield: 38 mg (31%) of methyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindole-5-carboxylate as a brown oil. HPLC / MS m / z: 430.20 [M+H] + , Rt (J): 0.53 min.

[0332] Example 49.4: A solution of intermediate 49.3 (96.9 mg, 0.225 mmol) and lithium hydroxide (22.8 mg, 0.952 mmol) in water (4.0 mL) and 1,4-dioxane (2.0 ml) was stirred at 60° C. overnight. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by RP flash chromatography to yield 75 mg (80%) of 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindole-5-carboxylic acid. HPLC / MS m / z: 416.25 [M+H] + , Rt (J): 0.52 min.

[0333] Example 49.5: The esterification was carried out as described in Intermediate 44.5. Yield: 9 mg (6%) of ethyl 2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindole-5-carboxylate as a yellow oil. HPLC / MS m / z: 444.20 [M+H] + , Rt (J): 0.58 min. 1 H NMR (300 MHz, DMSO-d6): δ 8.88 (d, J = 1.5 Hz, 1H), 8.17-8.08 (m, 1H), 7.96 (d, J = 5.8 Hz, 1H), 7.89 (t, J = 5.4 Hz, 1H), 7.86-7.76 (m, 3H), 7.37 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 5.7 Hz, 1H), 4.34-4.20 (m, 2H), 4.00 (s, 4H), 3.76-3.64 (m, 2H), 3.00 (t, J = 6.7 Hz, 2H), 2.68 (s, 3H), 1.30 (t, J = 7.1 Hz, 3H).

[0334] The following examples were obtained in a similar manner. Example 50: Ethyl 2-(2-{[7-(1-methyl-1H-pyrazol-4-yl)isoquinolin-1-yl]amino}ethyl)-2,3-dihydro-1H-isoindole-5-carboxylate [ka] 14 mg off-white solid. HPLC / MS m / z: 442.25 [M+H] + , Rt (J): 0.57 min. 1H NMR (300 MHz, DMSO-d6): δ 8.39 (d, J = 1.5 Hz, 1H), 8.19 (s, 1H), 8.00 (d, J = 0.8 Hz, 1H), 7.90-7.77 (m, 4H), 7.66 (d, J = 8.5 Hz, 1H), 7.39 (t, J = 8.0 Hz, 2H), 6.83 (d, J = 5.8 Hz, 1H), 4.28 (q, J = 7.1 Hz, 2H), 4.01 (s, 4H), 3.88 (s, 3H), 3.78-3.61 (m, 2H), 3.00 (t, J = 6.8 Hz, 2H), 1.30 (t, J = 7.1 Hz, 3H).

[0335] Example 51: Ethyl 1-methyl-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-indazole-5-carboxylate [ka]

[0336] Example 51.1: 7-(5-Methyl-1,2,4-oxadiazol-3-yl)isoquinolin-2-ium-2-olate (708.0 mg, 3.116 mmol) and aminoacetaldehyde diethyl acetal (519.0 mg, 3.897 mmol) were dissolved in dichloromethane (16.0 mL) under an argon atmosphere. N-Ethyldiisopropylamine (1.91 g, 14.778 mmol) and PyBroP (1.89 g, 4.054 mmol) were added, and the reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with dichloromethane and washed with water. The organic layer was dried over sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by flash chromatography to yield 681 mg (64%) of N-(2,2-diethoxyethyl)-7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-amine as a beige solid. HPLC / MS m / z: 342.9 [M+H] +, Rt (M): 1.67 min.

[0337] Example 51.2: Intermediate 51.1 (681.0 mg, 1.989 mmol) was dissolved in THF (5.0 mL), hydrochloric acid (2N, 10.0 mL) was added, and the reaction mixture was heated to 60° C. and stirred for 3 h. The reaction mixture was cooled to room temperature and made basic with 6N NaOH. A precipitate formed, which was filtered off, washed with water, and dried to give 468 mg (88%) of 2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}acetaldehyde as a beige solid. HPLC / MS m / z: 268.9 [M+H] + , Rt (M): 1.40 min.

[0338] Example 51.3: Intermediate 51.2 (448.0 mg, 1.670 mmol) was dissolved in methanol (11.0 mL), N-methyl(tert-butoxy)carbohydrazide (293.0 mg, 2.004 mmol) and glacial acetic acid (430 μL, 7.515 mmol) were added, and the reaction mixture was stirred at room temperature for 1 h. Sodium cyanoborohydride (157.4 mg, 2.505 mmol) was added slowly, and the reaction was stirred for 24 h. The reaction mixture was evaporated, and the residue was dissolved in dichloromethane and treated with aqueous saturated NaHCO3 solution. The mixture was stirred vigorously until gas formation ceased. The phases were separated, the aqueous phase was extracted with dichloromethane, and the combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure to yield 665 mg (100%) of N-methyl-N'-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)(tert-butoxy)carbohydrazide as a yellow oil. HPLC / MS m / z: 398.9 [M+H] + , Rt (M): 1.74 min.

[0339] Example 51.4: 2-Bromo-5-(methoxycarbonyl)benzoic acid (528.2 mg, 2.039 mmol) was dissolved in thionyl chloride (1.23 mL, 16.991 mmol) and stirred at 75° C. for 1 h. Excess thionyl chloride was removed in vacuo, and the residue was dissolved in a small amount of dichloromethane and added slowly to a solution of Intermediate 51.3 (677.0 mg, 1.699 mmol) and triethylamine (7.07 mL, 50.974 mmol) in dichloromethane (11.0 mL). The reaction mixture was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the residue was purified by RP flash chromatography to afford 826 mg (76%) of methyl 4-bromo-3-{N'-[(tert-butoxy)carbonyl]-N'-methyl-N-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)hydrazinecarbonyl}benzoate as a yellow oil. HPLC / MS m / z: 639.1 / 641.1 [M+H] + , Rt (N): 1.61 min.

[0340] Example 51.5: Intermediate 51.4 (826.0 mg, 1.292 mmol) was dissolved in a solution of HCl in 1,4-dioxane (4.0 M, 11.2 mL) and stirred at room temperature for 2.5 h, during which time a precipitate formed. The reaction mixture was concentrated in vacuo, and the residue was dissolved in dichloromethane and washed with aqueous saturated NaHCO3 solution. The organic phase was dried over sodium sulfate, filtered, and evaporated. The crude product (650 mg) was used in the next step without further purification. HPLC / MS m / z: 539.1 / 541.1 [M+H] + , Rt (N): 1.35 min.

[0341] Example 51.6: Intermediate 51.5 (650.0 mg, 1.205 mmol) was dissolved in DMF (3.50 mL), and cesium carbonate (784.8 mg, 2.409 mmol), copper iodide (22.9 mg, 0.120 mmol), and 2-acetylcyclohexanone (33.8 mg, 0.241 mmol) were added under an argon atmosphere. The reaction mixture was heated to 90 °C and stirred for 5 h. After cooling to room temperature, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by RP flash chromatography to give 442 mg (80%) of methyl 1-methyl-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-indazole-5-carboxylate as a colorless solid. HPLC / MS m / z: 459.1 [M+H] + , Rt (N): 1.25 min.

[0342] Example 51.7: Saponification of intermediate 51.6 (274.0 mg, 0.598) and esterification of the corresponding acid (50 mg, 0.090 mmol) were carried out as described in Example 3. Yield: 34 mg (80%) of ethyl 1-methyl-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-indazole-5-carboxylate, a colorless solid. HPLC / MS m / z: 472.8 [M+H] + , Rt (M): 1.72 min. 1H NMR (700 MHz, DMSO-d6): δ 8.78 (s, 1H), 8.18 (d, J = 1.7 Hz, 1H), 8.15 (dd, J = 8.3, 1.6 Hz, 1H), 8.09 (dd, J = 8.6, 1.7 Hz, 1H), 8.04 (t, J = 5.7 Hz, 1H), 7.96 (d, J = 5.7 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.57 (d, J = 8.7 Hz, 1H), 6.98 (d, J = 5.7 Hz, 1H), 4.30 (q, J = 7.1 Hz, 2H), 4.20 (t, J = 6.5 Hz, 2H), 3.73 (q, J = 6.3 Hz, 2H), 3.48 (s, 3H), 2.68 (s, 3H), 1.33 (t, J = 7.1 Hz, 3H).

[0343] The following examples were obtained in a similar manner. Example 52: Propan-2-yl 1-methyl-2-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-3-oxo-2,3-dihydro-1H-indazole-5-carboxylate [ka] 56 mg colorless solid. HPLC / MS m / z: 486.9 [M+H] + , Rt (M): 1.79 min. 1H NMR (700 MHz, DMSO-d6): δ 8.78 (s, 1H), 8.18-8.13 (m, 2H), 8.10-8.06 (m, 1H), 8.03 (t, J = 5.7 Hz, 1H), 7.96 (d, J = 5.7 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.57 (d, J = 8.6 Hz, 1H), 6.98 (d, J = 5.7 Hz, 1H), 5.12 (p, J = 6.2 Hz, 1H), 4.20 (t, J = 6.6 Hz, 2H), 3.73 (q, J = 6.3 Hz, 2H), 3.48 (s, 3H), 2.69 (s, 3H), 1.32 (d, J = 6.2 Hz, 6H).

[0344] Example 53: 1-[6-(2-{[7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino}ethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridin-2-yl]butan-1-one [ka]

[0345] Example 53.1: Ethyl 4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate hydrochloride (33.2 mg, 0.157 mmol) and 2-(Boc-amino)ethyl bromide (70.6 mg, 0.315 mmol) were placed in a vial and suspended in dry 1,4-dioxane (0.50 mL). Triethylamine (76.4 μL, 0.551 mmol) was added, and the mixture was heated to 80 °C and stirred for 18 h. The reaction mixture was cooled to room temperature and diluted with 5 mL of dichloromethane and 5 mL of water. The aqueous phase was washed twice with dichloromethane, and the combined organic layers were dried over sodium sulfate, filtered, and evaporated to dryness. The residue was purified by flash chromatography to give 45 mg (81%) of ethyl 6-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate as a yellow oil. HPLC / MS m / z: 354.9 [M+H] + , Rt (H): 0.70 min.

[0346] Example 53.2: Intermediate 53.1 (157.4 mg, 0.444 mmol) was dissolved in dry THF (7 mL) and water (4 mL). With stirring, lithium hydroxide (26.6 mg, 1.110 mmol) was added and the reaction mixture was stirred at room temperature overnight. A pale yellow / orange solution formed, which was stirred at room temperature for an additional 48 h. The reaction mixture was diluted with water (40 mL) and acidified to pH 3-4 with 1.0 N HCl solution. The mixture was extracted twice with ethyl acetate, and the combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The aqueous phase was neutralized with saturated aqueous NaHCO3 solution and evaporated to dryness. The colorless solid was combined with the solid isolated after extraction, triturated with a mixture of dichloromethane / methanol / dimethylformamide, filtered, washed, and the filtrate evaporated to dryness to give 145 mg (100%) of 6-(2-{[(tert-butoxy)carbonyl]amino}ethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylic acid as a beige solid. HPLC / MS m / z: 326.9 [M+H] +, Rt (H): 0.63 min.

[0347] Example 53.3: Intermediate 53.2 (145.0 mg, 0.444 mmol) and N,O-dimethylhydroxylamine hydrochloride (52.0 mg, 0.533 mmol) were suspended in N,N-dimethylformamide (1.6 mL). [Dimethylamino-([1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methylene]dimethylammonium; hexafluorophosphate (HATU) (168.8 mg, 0.444 mmol) and N-ethyldiisopropylamine (0.38 mL, 2.221 mmol) were added, and the mixture was stirred at room temperature for 2 h. Further N,O-dimethylhydroxylamine (21.7 mg, 0.222 mmol) and [dimethylamino-([1,2,3]triazolo[4,5-b]pyridin-3-yloxy)methylene]dimethylammonium; hexafluorophosphate (HATU) (84.4 mg, 0.222 mmol) were added, and the reaction was stirred at room temperature overnight. The mixture was diluted with ethyl acetate and extracted twice with water and saturated aqueous NaHCO solution (1:1). The combined aqueous phases were extracted twice with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to yield 121 mg (73%) of tert-butyl N-(2-{2-[methoxy(methyl)carbamoyl]-4H,5H,6H,7H-thieno[2,3-c]pyridin-6-yl}ethyl)carbamate as a beige oil. HPLC / MS m / z: 370.2 [M+H] + , Rt (N): 1.15 min.

[0348] Example 53.4: tert-Butyl N-(2-{2-[methoxy(methyl)carbamoyl]-4H,5H,6H,7H-thieno[2,3-c]pyridin-6-yl}ethyl)carbamate (119.7 mg, 0.324 mmol) was dissolved in dry THF (0.50 mL). The clear solution was cooled to 0° C. in an ice bath. Bromo(propyl)magnesium (3 M solution, 119 μl, 0.357 mmol) was added slowly and the yellow solution was allowed to warm to room temperature overnight. Further bromo(propyl)magnesium (119 μl, 0.357 mmol) was added, and the reaction mixture was stirred at room temperature. After 30 min, a thick yellow gum precipitated at the bottom of the flask. Dry THF (0.50 mL) was added, and the mixture was stirred at room temperature overnight. The reaction was quenched with saturated aqueous NH4Cl solution and extracted twice using ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to afford 61 mg (53%) of tert-butyl N-(2-{2-butanoyl-4H,5H,6H,7H-thieno[2,3-c]pyridin-6-yl}ethyl)carbamate as a yellow oil. HPLC / MS m / z: 352.9 [M+H] + , Rt (H): 0.71 min.

[0349] Example 53.5: Deprotection of intermediate 53.4 (61 mg, 0.172 mmol) with HCl in 1,4-dioxane was carried out as described for compound 25.7 to yield 43.4 mg (100%) of 1-[6-(2-aminoethyl)-4H,5H,6H,7H-thieno[2,3-c]pyridin-2-yl]butan-1-one as a brown oil, which was used in the next step without further purification.

[0350] Example 54.6: Prepared as described in Example 32.3. Yield: 21 mg of an orange solid. HPLC / MS m / z: 461.9 [M+H] + , Rt (M): 1.76 min. 1H NMR (400 MHz, DMSO-d6): δ 8.87-8.85 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 7.96 (d, J = 5.8 Hz, 1H), 7.83 (d, J = 8.4 Hz, 1H), 7.84-7.80 (m, 1H), 7.67 (s, 1H), 6.94-6.92 (m, 1H), 3.81-3.78 (m, 2H), 3.71 (q, J = 6.4 Hz, 2H), 2.87-2.79 (m, 6H), 2.70 (s, 3H), 2.72-2.66 (m, 2H), 1.67-1.56 (m, 2H), 0.91 (t, J = 7.4 Hz, 3H).

[0351] Example 54: Ethyl 6-methyl-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate [ka]

[0352] Example 54.1: Ethyl pyruvate (0.67 mL, 6.0282 mmol) and ethylenediamine (0.40 mL, 6.0282 mmol) were added to MeCN (10.05 mL, 0.3700 M) and stirred vigorously for 30 min at 20° C. Ethyl 3-bromo-2-oxobutanoate (1260.14 mg, 6.0282 mmol) and iron(III) chloride (195.56 mg, 1.2056 mmol) in MeCN (6.03 mL, 0.3700 M) were added to the mixture and heated to reflux for 5 h. After completion of the reaction as indicated by LCMS, the mixture was cooled to room temperature. The solution was passed through Celite to remove residual FeO, then the organics were evaporated and the crude material was subjected to normal phase column chromatography (0-100% EtOAc:CycHex → 10% MeOH) to afford crude ethyl 6-methyl-1-oxo-3,4-dihydro-2H-pyrrolo[1,2-a]pyrazine-7-carboxylate (567.5 mg, 42%, 2.5535 mmol) as a dark brown solid. The product was used in the next step without further purification. HPLC / MS m / z 223.1078 [M+H] + , Rt (AI): 1.05 min.

[0353] Example 54.2: To a solution of ethyl 6-methyl-1-oxo-3,4-dihydro-2H-pyrrolo[1,2-a]pyrazine-7-carboxylate (550.00 mg, 2.4748 mmol) in THF (24.75 mL, 0.1000 M) at 0° C. was added sodium hydride (60% dispersion in mineral oil) (217.78 mg, 5.4446 mmol), and the mixture was stirred and allowed to warm to room temperature over 1 hour. Bromoacetonitrile (0.57 mL, 8.1668 mmol) was then added, and the reaction was allowed to stir at room temperature overnight. The reaction was quenched with NH4Cl, followed by the addition of DCM, and the reaction was passed through a phase separator and washed several times with DCM. The combined organic layers were evaporated in vacuo to give the crude product, which was subjected to silica gel normal phase column chromatography (40-100% EtOAc:CycHex) to afford ethyl 2-(cyanomethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (142 mg, 22%, 0.5435 mmol) as a light yellow / brown solid. HPLC / MS m / z 262.1195 [M+H] + , m / z 284.0994 [M+Na] + , Rt (AC): 1.06 min. 1 H NMR (600 MHz, Chloroform-d): δ 7.40 (s, 1H), 4.53 (s, 2H), 4.28 (q, J = 7.1 Hz, 2H), 4.17-4.12 (m, 2H), 3.86-3.79 (m, 2H), 2.56 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0354] Example 54.3: To a solution of ethyl 2-(cyanomethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (140.00 mg, 0.5358 mmol) in EtOH (8.00 mL, 0.0500 M) was added AcOH (0.15 mL, 0.0500 M) and water (2.50 mL, 0.0500 M). The reaction mixture was placed under an atmosphere of H2, and the solution was stirred at room temperature for 16 h. The Pd / C was filtered through a pad of Celite and washed with MeOH (5 mL). The crude sample was evaporated and subjected to reverse-phase column chromatography (10-80% MeOH:water). The pure fractions were passed through an SCX-II column and released using 2N NH3 in methanol, yielding ethyl 2-(2-aminoethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (80 mg, 56%, 0.3015 mmol) as a pale yellow semi-solid. HPLC / MS m / z 249.1238 [M+H] + , Rt (AC): 0.66 min. 1 H NMR (600 MHz, DMSO-d6): δ 6.90 (s, 1H), 4.18 (q, J = 7.1 Hz, 2H), 4.12-4.05 (m, 2H), 3.73-3.65 (m, 2H), 3.46 (t, J = 6.5 Hz, 2H), 2.78 (t, J = 6.5 Hz, 2H), 2.47 (s, 3H), 1.26 (t, J = 7.1 Hz, 3H).

[0355] Example 54.4: 5-Methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (43.16 mg, 0.1900 mmol), DIPEA (0.10 mL, 0.5936 mmol), and bromotri(pyrrolidino)phosphonium hexafluorophosphate (88.56 mg, 0.1900 mmol) in anhydrous DCM (0.75 mL, 0.1300 M) were stirred at 40° C. in a microwave vial under a nitrogen atmosphere for 30 min. Ethyl 2-(2-aminoethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (42.00 mg, 0.1583 mmol) was then added to anhydrous DCM (0.50 mL, 0.1300 M), and the reaction mixture was heated at 60 °C for 2 h by microwave irradiation. Volatiles were removed under reduced pressure. The crude material was then directly purified by reverse-phase flash chromatography (10-80% MeOH in water). One fraction was clean by LCMS and was passed through an SCX-II column to yield the clean product ethyl 6-methyl-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (20.3 mg, 27%, 0.0421 mmol). The other fractions were washed through an SCX-II column from which further material was collected and evaporated to afford ethyl 6-methyl-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (11.7 mg, 15%, 0.0242 mmol) as a fine, 98% pure, pale yellow solid which was used in the next reaction. HPLC / MS m / z 475.2073 [M+H] + , Rt (AD): 2.16 min. 1H NMR (600 MHz, DMSO-d6): δ 8.89-8.86 (m, 1H), 8.15 (dd, J = 8.5, 1.5 Hz, 1H), 8.04 (t, J = 5.2 Hz, 1H), 7.98 (d, J = 5.7 Hz, 1H), 7.85 (d, J = 8.5 Hz, 1H), 6.97 (d, J = 5.7 Hz, 1H), 6.91 (s, 1H), 4.17 (q, J = 7.1 Hz, 2H), 4.08-4.03 (m, 2H), 3.78-3.69 (m, 6H), 2.70 (s, 3H), 2.44 (s, 3H), 1.26 (t, J = 7.1 Hz, 3H).

[0356] Example 55: 6-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-2-propoxy-5H-pyrrolo[3,4-b]pyridin-7-one [ka]

[0357] Example 55.1: Methyl 6-chloro-3-methylpyridine-2-carboxylate (1000.00 mg, 5.3876 mmol) was dissolved in anhydrous DCE (53.88 mL, 0.1000 M) under argon. AIBN (88.47 mg, 0.5388 mmol) and NBS (958.89 mg, 5.3876 mmol) were added, and the reaction mixture was heated to reflux for 2 h. The reaction mixture was cooled to ambient temperature, evaporated onto silica gel, and purified by silica gel normal phase chromatography (0-30% EtOAc in cyclohexane) to give methyl 3-(bromomethyl)-6-chloro-pyridine-2-carboxylate (1.12 g, 79%, 4.2457 mmol) as a colorless crystalline solid. HPLC / MS m / z 287.9862 [M+Na] + , Rt (AJ): 2.07 min. 1H NMR (600 MHz, DMSO-d6): δ 8.14 (d, J = 8.3 Hz, 1H). 7.78 (d, J = 8.3 Hz, 1H), 4.94 (s, 2H), 3.91 (s, 3H).

[0358] Example 55.2: Methyl 3-(bromomethyl)-6-chloropyridine-2-carboxylate (1.12 g, 3.3875 mmol), DIPEA (1.18 mL, 6.775 mmol), and 1-Boc-ethylenediamine (0.56 mL, 3.5569 mmol) were mixed in anhydrous MeCN (33.88 mL, 0.1000 M) under argon and heated to reflux for 1.5 h. The reaction mixture was cooled to ambient temperature, evaporated onto silica gel, and purified by silica gel normal-phase flash chromatography (0–100% EtOAc in cyclohexane) to give tert-butyl N-[2-(2-chloro-7-oxo-5H-pyrrolo[3,4-b]pyridin-6-yl)ethyl]carbamate (748 mg, 71%, 2.3993 mmol) as a colorless crystalline solid. HPLC / MS m / z 334.1697 [M+Na] + , Rt(AJ): 1.99 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.15 (d, J = 8.1 Hz, 1H), 7.68 (d, J = 8.1 Hz, 1H), 6.94 (t, J = 6.1 Hz, 1H), 4.51 (s, 2H), 3.58 (t, J = 5.9 Hz, 2H), 3.18-3.23 (m, 2H), 1.27 (s, 9H).

[0359] Example 55.3: tert-Butyl N-[2-(2-chloro-7-oxo-5H-pyrrolo[3,4-b]pyridin-6-yl)ethyl]carbamate (150.00 mg, 0.4811 mmol), cesium carbonate (220.83 mg, 0.6736 mmol), tBuBrettPhos Pd G3 (17.13 mg, 0.0192 mmol), and 1-propanol (0.05 mL, 0.6255 mmol) were suspended in 1,4-dioxane (1.20 mL, 0.4000 M) under N2. The mixture was then evacuated and filled with N2 (x5, 15 sec evacuation). The mixture was then heated to 80 °C and stirred (1200 RPM) under N2. The reaction was left stirring overnight. The reaction mixture was dried under reduced pressure and subjected to silica gel normal phase column chromatography (0-100% EtOAc:CycHex). Fractions were combined and evaporated to give tert-butyl N-[2-(7-oxo-2-propoxy-5H-pyrrolo[3,4-b]pyridin-6-yl)ethyl]carbamate (137 mg, 85%, 0.4085 mmol) as a clear viscous oil. HPLC / MS m / z 358.1727 [M+H] + , Rt (AI): 1.42 min. 1 H NMR (600 MHz, Chloroform-d): δ 7.68 (d, J = 8.4 Hz, 1H), 6.90 (d, J = 8.4 Hz, 1H), 4.99 (s, 1H), 4.44 (t, J = 6.8 Hz, 2H), 4.40 (s, 2H), 3.79 (t, J = 6.0 Hz, 2H), 3.47 (q, J = 6.0 Hz, 2H), 1.83 (h, J = 7.2 Hz, 2H), 1.36 (s, 9H), 1.05 (t, J = 7.4 Hz, 3H).

[0360] Example 55.4: tert-Butyl N-[2-(7-oxo-2-propoxy-5H-pyrrolo[3,4-b]pyridin-6-yl)ethyl]carbamate (130.00 mg, 0.3876 mmol) was mixed with 4 M HCl in dioxane (9.69 mL, 38.76 mmol) and anhydrous 1,4-dioxane (10.00 mL, 0.0400 M) at room temperature under argon and stirred overnight. Volatiles were removed under reduced pressure. The crude material was dissolved in MeOH / water and filtered through a 1 g SCX column. The product was released with 2N ammonia in MeOH to give 6-(2-aminoethyl)-2-propoxy-5H-pyrrolo[3,4-b]pyridin-7-one (74.1 mg, 81%, 0.3149 mmol) as an off-white solid. HPLC / MS m / z 258.1211 [M+H] + , Rt (AI): 0.79 min.

[0361] Example 55.5: 6-(2-aminoethyl)-2-propoxy-5H-pyrrolo[3,4-b]pyridin-7-one (30.00 mg, 0.1275 mmol), 5-methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (34.77 mg, 0.1530 mmol), PyBrop (71.33 mg, 0.1530 mmol), N,N-diisopropylethylamine (0.08 mL, 0.4782 mmol), and anhydrous DCM (0.64 mL, 0.2000 M) were placed in a microwave vial under nitrogen at room temperature. The reaction mixture was heated at 60 °C by microwave irradiation for 1 h. The volatiles were removed under reduced pressure. The crude material was directly purified by reverse-phase flash chromatography (10-60% MeOH in water). The product-containing fractions were filtered through a 1 g SCX column. The product was released with ammonia in MeOH to give 6-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-2-propoxy-5H-pyrrolo[3,4-b]pyridin-7-one (15.3 mg, 27%, 0.0344 mmol) as an amorphous solid. HPLC / MS m / z 445.1993 [M+H]+ , Rt (AJ): 2.14 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.83 (d, J = 1.6 Hz, 1H), 8.14 (dd, J = 8.4, 1.5 Hz, 1H), 8.03 (t, J = 5.6 Hz, 1H), 7.93 (dd, J = 13.3, 7.0 Hz, 2H), 7.84 (d, J = 8.5 Hz, 1H), 6.98-6.93 (m, 2H), 4.48 (s, 2H), 4.23 (t, J = 6.7 Hz, 2H), 3.85 (t, J = 5.8 Hz, 2H), 3.79 (q, J = 5.7 Hz, 2H), 2.68 (s, 3H), 1.72 (h, J = 7.1 Hz, 2H), 0.96 (t, J = 7.4 Hz, 3H).

[0362] Example 56: 6-(5-methyl-1,3,4-oxadiazol-2-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one [ka]

[0363] Example 56.1: A solution of methyl 5-bromo-2-(bromomethyl)benzoate (3000.00 mg, 9.7412 mmol), 1-Boc-ethylenediamine (1794.73 mg, 11.202 mmol), and triethylamine (2.06 mL, 14.612 mmol) in MeOH (50.00 mL, 0.1900 M) was refluxed under nitrogen overnight. After cooling to room temperature, the reaction mixture was concentrated in vacuo to give the crude product, which was purified by silica gel normal phase column chromatography (eluting with 20-60% EtOAc:CycHex) to give tert-butyl N-[2-(6-bromo-1-oxo-isoindolin-2-yl)ethyl]carbamate (2372.5 mg, 69%, 6.6788 mmol) as a white solid. HPLC / MS m / z 377.037, 379.035 [M+Na] + , Rt (AI): 1.40 min. 1 H NMR (600 MHz, Methanol-d4): δ 7.88 (d, J = 1.9 Hz, 1H), 7.74 (dd, J = 8.1, 1.9 Hz, 1H), 7.50 (d, J = 8.0 Hz, 1H), 4.54 (s, 2H), 3.69 (dd, J = 6.5, 5.1 Hz, 2H), 3.35 (dd, J = 6.5, 5.1 Hz, 2H), 1.31 (s, 9H).

[0364] Example 56.2: tert-Butyl N-[2-(6-bromo-1-oxo-isoindolin-2-yl)ethyl]carbamate (400.00 mg, 1.126 mmol) and Xantphos Palladacycle Gen. 4 (21.70 mg, 0.0225 mmol) were combined in a 5 mL microwave vial. The vial was capped, flushed with argon, and evacuated before being fitted with a balloon filled with carbon monoxide (63.08 mg, 2.2521 mmol). Anhydrous 1,4-dioxane (2.25 mL, 0.5000 M) and acetohydrazide (150.15 mg, 2.0269 mmol) were added, and the reaction mixture was heated at 90° C. overnight. To this crude product was added N,N-diisopropylethylamine (0.20 mL, 1.126 mmol) and p-toluenesulfonyl chloride (214.67 mg, 1.126 mmol). After 2 h, another portion of p-toluenesulfonyl chloride (214.67 mg, 1.126 mmol) was added, and the reaction was stirred overnight. The crude mixture was subjected to silica gel normal phase column chromatography (0-100% EtOAc:CycHex → 10% MeOH:EtOAc) to afford tert-butyl N-[2-[6-(5-methyl-1,3,4-oxadiazol-2-yl)-1-oxo-isoindolin-2-yl]ethyl]carbamate (195.6 mg, 48%, 0.5458 mmol) as a fine cream-colored solid. HPLC / MS m / z 381.1517 [M+Na] + , Rt (AI): 1.22 min. 1 H NMR (600 MHz, Chloroform-d): δ 8.42 (d, J = 1.5 Hz, 1H), 8.29 (dd, J = 8.0, 1.6 Hz, 1H), 7.60 (dd, J = 7.8, 1.0 Hz, 1H), 4.88 (d, J = 7.1 Hz, 1H), 4.57 (s, 2H), 3.77 (t, J = 6.0 Hz, 2H), 3.46 (q, J = 6.0 Hz, 2H), 2.64 (s, 3H), 1.31 (s, 9H).

[0365] Example 56.3: tert-Butyl N-[2-[6-(5-methyl-1,3,4-oxadiazol-2-yl)-1-oxo-isoindolin-2-yl]ethyl]carbamate (150.00 mg, 0.4185 mmol) was mixed with 4 M HCl in dioxane (10.46 mL, 41.854 mmol) and anhydrous 1,4-dioxane (10.00 mL, 0.0400 M) at room temperature under argon and stirred for 2 h. Volatiles were removed under reduced pressure. The crude material was dissolved in MeOH / water and filtered through a 2 g SCX column. The product was released with 2M ammonia in MeOH to give 2-(2-aminoethyl)-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoindolin-1-one (93 mg, 86%, 0.3601 mmol) as an off-white solid. HPLC / MS m / z 259.1211 [M+H] + , Rt (AI): 0.61 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.19 (dd, J = 7.9, 1.6 Hz, 1H), 8.14 (d, J = 1.5 Hz, 1H), 7.86-7.80 (m, 1H), 4.65 (s, 2H), 3.54 (t, J = 6.4 Hz, 2H), 2.82 (t, J = 6.4 Hz, 2H), 2.61 (s, 3H).

[0366] Example 56.4: 2-(2-aminoethyl)-6-(5-methyl-1,3,4-oxadiazol-2-yl)isoindolin-1-one (45.00 mg, 0.1742 mmol), 5-methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (47.51 mg, 0.209 mmol), PyBrop (97.47 mg, 0.2091 mmol), N,N-diisopropylethylamine (0.11 mL, 0.6534 mmol), and anhydrous DCM (0.87 mL, 0.2000 M) were placed in a microwave vial under nitrogen at room temperature. The reaction mixture was heated at 60 °C by microwave irradiation for 2 h. The volatiles were removed under reduced pressure. The crude material was directly purified by reverse-phase flash chromatography (10–60% MeOH in water). Because all fractions contained impurities, all product-containing fractions were filtered through a 2 g SCX column, the product was released with ammonia in MeOH, and the solvent was evaporated in vacuo. The crude material was then subjected to silica gel normal-phase column chromatography (20–100% EtOAc:CycHex → 10% MeOH:EtOAc). Fractions containing pure product were collected and evaporated in vacuo to yield 6-(5-methyl-1,3,4-oxadiazol-2-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one (19.3 mg, 24%, 0.0413 mmol) as a fine white solid. HPLC / MS m / z 468.1758 [M+H] + , Rt (AJ): 1.84 min. 1H NMR (600 MHz, DMSO-d6): δ 8.85-8.81 (m, 1H), 8.18 (dd, J = 7.9, 1.6 Hz, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.10 (dd, J = 1.5, 0.7 Hz, 1H), 8.04 (t, J = 5.6 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.87-7.80 (m, 2H), 6.95 (dd, J = 5.9, 0.8 Hz, 1H), 4.70 (s, 2H), 3.88 (dd, J = 6.6, 5.1 Hz, 2H), 3.82 (q, J = 5.6 Hz, 2H), 2.67 (s, 3H), 2.59 (s, 3H).

[0367] Example 57: Propan-2-yl 6-methyl-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate [ka]

[0368] Example 57.1: N-Iodosuccinimide (3493.46 mg, 15.668 mmol) was added slowly in portions to a solution of ethyl 5-methyl-1H-pyrrole-2-carboxylate (2400.00 mg, 15.668 mmol) in anhydrous DCM (31.34 mL, 0.5000 M) at 0 °C. The reaction was stirred at 0 °C for 2 h, then allowed to warm to room temperature and stirred overnight. The reaction mixture was concentrated, and the crude mixture was separated by silica gel normal phase column chromatography (0-20% EtOAc:CycHex) to afford ethyl 4-iodo-5-methyl-1H-pyrrole-2-carboxylate (4.12 g, 94%, 14.766 mmol) as a fine white solid. HPLC / MS m / z 251.9519 [M+H-CH2CH3] + , Rt (AC): 1.36 min. 1H NMR (600 MHz, Chloroform-d): δ 9.29 (s, 1H), 6.93 (d, J = 2.6 Hz, 1H), 4.30 (q, J = 7.1 Hz, 2H), 2.30 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0369] Example 57.2: A solution of ethyl 4-iodo-5-methyl-1H-pyrrole-2-carboxylate (4000.00 mg, 14.333 mmol), tert-butyl-N-hydroxyethylcarbamate (3.99 mL, 25.8 mmol), and triphenylphosphine (6015.06 mg, 22.933 mmol) in anhydrous THF (35.83 mL, 0.4000 M) was cooled to 0° C. Diisopropyl azodicarboxylate (8.47 mL, 43 mmol) was added dropwise over 20 min, and the resulting mixture was allowed to warm to room temperature and stirred for 18 h. The crude product was evaporated under reduced pressure and subjected to normal phase column chromatography (0-10% EtOAc:Cychex) to afford impure ethyl 1-[2-(tert-butoxycarbonylamino)ethyl]-4-iodo-5-methylpyrrole-2-carboxylate (3.65 g, 57%, 8.2127 mmol) as a cream-colored solid, which was used in the next step without further purification. HPLC / MS m / z 445.0591 [M+Na] + , Rt (AI): 1.69 min. 1 H NMR (500 MHz, Chloroform-d): δ 7.07 (s, 1H), 4.43 (t, J = 6.2 Hz, 2H), 4.26 (dq, J = 21.4, 7.1 Hz, 2H), 3.40 (q, J = 6.1 Hz, 2H), 2.30 (d, J = 7.2 Hz, 3H), 1.41 (s, 9H), 1.37-1.26 (m, 3H).

[0370] Example 57.3: Ethyl 1-[2-(tert-butoxycarbonylamino)ethyl]-4-iodo-5-methylpyrrole-2-carboxylate (2.85 g, 6.7494 mmol) was mixed with 1,4-dioxane (67.49 mL, 269.98 mmol) and 4 N HCl in anhydrous 1,4-dioxane (67.53 mL, 0.0700 M) at room temperature under argon and stirred for 2 h. The volatiles were removed under reduced pressure. The crude material was dissolved in EtOH (26.87 mL, 0.0700 M) and triethylamine (4.75 mL, 33.747 mmol), and the mixture was stirred at 80 °C for 72 h. The crude mixture was evaporated under reduced pressure, adsorbed onto silica, and subjected to silica gel normal phase column chromatography (0-10% MeOH:DCM). The fractions were collected and evaporated to give 7-iodo-6-methyl-3,4-dihydro-2H-pyrrolo[1,2-a]pyrazin-1-one (1.34 g, 72%, 4.8372 mmol) as a cream solid. HPLC / MS m / z 276.9833 [M+H] + , Rt (AI): 1.14 min.

[0371] Example 57.4: 7-Iodo-6-methyl-3,4-dihydro-2H-pyrrolo[1,2-a]pyrazin-1-one (700.00 mg, 2.5356 mmol) and Xantphos Palladacycle Gen. 4 (146.56 mg, 0.1521 mmol) were combined in a 20 mL microwave vial. The vial was capped, flushed with argon, and evacuated. Afterwards, a balloon filled with carbon monoxide (717.39 mg, 25.356 mmol) was attached, and anhydrous 1,4-dioxane (8.45 mL, 0.3000 M) and 2-propanol (9.71 mL, 126.78 mmol) were added. The reaction mixture was heated at 80° C. overnight. The reaction was evaporated under reduced pressure and the crude material was subjected to silica gel normal phase column chromatography (0-10% DCM:MeOH) to afford semi-pure propan-2-yl 6-methyl-1-oxo-3,4-dihydro-2H-pyrrolo[1,2-a]pyrazine-7-carboxylate (300 mg, 50%, 1.2697 mmol) as a dark brown amorphous solid. HPLC / MS m / z 237.1241 [M+H] + , Rt (AI): 1.13 min. The product was used in the next step without further purification.

[0372] Example 57.5: To a solution of propan-2-yl 6-methyl-1-oxo-3,4-dihydro-2H-pyrrolo[1,2-a]pyrazine-7-carboxylate (360.00 mg, 1.5237 mmol) in THF (15.24 mL, 0.1000 M) at 0 °C, NaH (134.08 mg, 3.3521 mmol) was added, and the mixture was stirred and allowed to warm to room temperature over 1 h. Bromoacetonitrile (0.35 mL, 5.0281 mmol) was then added, and the reaction was allowed to stir at room temperature overnight. The crude material was evaporated and subjected to silica gel normal phase column chromatography (0-10% MeOH:DCM). The pure fractions were collected and evaporated to give propan-2-yl 2-(cyanomethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (165.4 mg, 39%, 0.6008 mmol) as a pale yellow solid. HPLC / MS m / z 276.1347 [M+H] + , Rt (AI): 1.17 min. 1 H NMR (500 MHz, DMSO-d6): δ 7.00 (s, 1H), 5.03 (p, J = 6.2 Hz, 1H), 4.55 (s, 2H), 4.19-4.13 (m, 2H), 3.83-3.74 (m, 2H), 2.47 (s, 3H), 1.26 (d, J = 6.3 Hz, 6H).

[0373] Example 57.6: To a cold (0 °C) solution of propan-2-yl 2-(cyanomethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (150.00 mg, 0.5449 mmol) and cobalt(II) chloride hexahydrate (259.28 mg, 1.0897 mmol) in MeOH (10.90 mL, 0.0500 M) was added NaBH (288.07 mg, 5.4486 mmol) (portionwise over 10 min). The mixture was poured into hydrochloric acid (1 N, 15 mL) and stirred until the black precipitate dissolved. The aqueous layer was made alkaline with ammonia, extracted with CHCl3, dried, filtered, and concentrated to give propan-2-yl 2-(2-aminoethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (135.5 mg, 89%, 0.4851 mmol) as a yellow oil. HPLC / MS m / z 280.1659 [M+H] + , Rt (AI): 0.90 min.

[0374] Example 57.7: 5-Methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (87.85 mg, 0.3866 mmol), bromotri(pyrrolidino)phosphonium hexafluorophosphate (180.24 mg, 0.3866 mmol), and DIPEA (0.21 mL, 1.2082 mmol) in DCM (1.91 mL, 0.1200 M) were stirred at 40° C. in a microwave vial under a nitrogen atmosphere for 30 min. Propan-2-yl 2-(2-aminoethyl)-6-methyl-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (90.00 mg, 0.3222 mmol) was then added to DCM (0.77 mL, 0.1200 M), and the reaction was heated at 60 °C by microwave irradiation for 2 h. Volatiles were removed under reduced pressure. The crude product was directly purified by reverse-phase flash chromatography (10-80% MeOH in water). Fractions were combined, passed through an SCX-II column, and subjected to another reverse-phase flash chromatography (10-80% MeOH in water). Fractions that remained impure were evaporated and subjected to a third reverse-phase flash chromatography (10-80% MeOH in water) on the SCX-II. The pure fractions were passed through an SCX-II column, released with NH3 in water, and evaporated to give the pure product propan-2-yl 6-methyl-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1-oxo-3,4-dihydropyrrolo[1,2-a]pyrazine-7-carboxylate (15.01 mg, 9%, 0.0305 mmol) as a pale yellow solid (99.3% pure by UV). HPLC / MS m / z 489.2237 [M+H] + , Rt (AJ): 2.19 min. 1H NMR (600 MHz, DMSO-d6): δ 8.90-8.85 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.03 (t, J = 5.2 Hz, 1H), 7.97 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.96 (d, J = 5.7 Hz, 1H), 6.89 (s, 1H), 5.00 (hept, J = 6.3 Hz, 1H), 4.07-3.99 (m, 2H), 3.78-3.68 (m, 6H), 2.69 (s, 3H), 2.43 (s, 3H), 1.24 (d, J = 6.2 Hz, 6H).

[0375] Example 58: N-[2-[2-(ethoxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridin-6-yl]ethyl]-7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-amine [ka]

[0376] Example 58.1: To a solution of 6-(tert-butoxycarbonyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-carboxylic acid (250.00 mg, 0.8823 mmol) in THF (3.68 mL) was added dropwise a solution of 1 M lithium aluminum hydride in THF (0.97 mL, 0.9706 mmol) under a nitrogen atmosphere. The reaction was cooled to 0 °C and stirred for 15 min, then warmed to room temperature and stirred for an additional 1 h. The reaction mixture was carefully quenched with 40 μL of water, followed by 40 μL of 15% aqueous NaOH and 120 μL of water. After the quenching procedure, the reaction mixture was diluted with diethyl ether (~3 mL) and some anhydrous sodium sulfate was added. The mixture was stirred for 15 min before being filtered through a Telso phase separator, washing with diethyl ether and DCM. The resulting filtrate was evaporated to dryness to give the crude product. The crude material was dissolved in EtOAc (10 mL) and washed twice with HCl 0.5 M (5 mL). The organic layer was dried over Na2SO4 and evaporated to give tert-butyl 2-(hydroxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-6-carboxylate (133 mg, 45%, 0.3950 mmol) as a slightly yellow oil. HPLC / MS m / z 292.0849 [M+Na] + , Rt (AI): 1.38 min. 1 H NMR (600 MHz, Chloroform-d): δ 6.69 (d, J = 5.6 Hz, 1H), 4.74 (d, J = 5.5 Hz, 2H), 4.57 (d, J = 5.3 Hz, 2H), 3.65 (t, J = 7.0 Hz, 2H), 2.67-2.62 (m, 2H), 1.47 (d, J = 3.1 Hz, 9H).

[0377] Example 58.2: To a solution of tert-butyl 2-(hydroxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-6-carboxylate (144.44 mg, 0.4826 mmol) in DMF (0.90 mL) was added sodium hydride (21.24 mg, 0.5309 mmol) under ice-cooling (the colorless solution turned yellow). After stirring at 0 °C for 5 min, iodoethane (0.04 mL, 0.5309 mmol) was added and stirring was continued for an additional 4 h. The reaction mixture was partitioned between water (20 mL) and ethyl acetate (30 mL). After phase separation, the aqueous layer was further extracted with ethyl acetate (2 × 30 mL). The combined organic layers were washed with brine (40 mL), dried (MgSO), filtered, and evaporated to dryness. The resulting crude material (79.8 mg) was purified by normal-phase silica column chromatography using a 10 g KP-silica column and a gradient of 0-50% EtOAc in cyclohexane as eluent. Fractions were combined and evaporated to dryness to give the required product, tert-butyl 2-(ethoxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-6-carboxylate (87 mg, 61%, 0.2925 mmol), as a clear oil. HPLC / MS m / z 320.1118 [M+Na] + , Rt (AI): 1.59 min. 1 H NMR (600 MHz, Chloroform-d): δ 6.67 (s, 1H), 4.56 (d, J = 5.6 Hz, 4H), 3.69-3.61 (m, 2H), 3.53 (q, J = 7.0 Hz, 2H), 2.64 (t, J = 6.1 Hz, 2H), 1.47 (s, 9H), 1.22 (t, J = 7.0 Hz, 3H).

[0378] Example 58.3: tert-Butyl 2-(ethoxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-6-carboxylate (87.00 mg, 0.2925 mmol) was dissolved in dry 1,4-dioxane (1.54 mL). Hydrogen chloride (4 M in 1,4-dioxane) (1.10 mL, 4.3879 mmol) was added and stirred at room temperature overnight. Volatiles were evaporated to dryness. The resulting salt, 2-(ethoxymethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine hydrochloride (67 mg, 49%, 0.1439 mmol) (beige powder), was carried on to the next step without further purification. HPLC / MS m / z 198.0871 [M+H] + , Rt (AI): 0.52 min.

[0379] Example 58.4: 2-(Ethoxymethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine hydrochloride (81.00 mg, 0.3465 mmol) was dissolved in CHCl3 (1.73 mL). Triethylamine (0.05 mL, 0.3465 mmol) was added, and the reaction mixture was cooled to 0 °C. A 1 M solution of 1-nitroethene in xylene (0.35 mL, 0.3465 mmol) was added, and the reaction was stirred at 0 °C for 10 min. The reaction was then stirred at room temperature for 1 h 40 min. The reaction mixture was concentrated in vacuo. The resulting crude product (53.5 mg) was purified by silica gel column chromatography using a Biotage KP-NH 12 g column (eluent: 20-80% EtOAc in cyclohexane). The combined fractions were evaporated to dryness to give a colorless oil, 2-(ethoxymethyl)-6-(2-nitroethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine (36 mg, 37%, 0.1265 mmol), as a white solid. HPLC / MS m / z 271.1031 [M+H] + , Rt (AI): 0.63 min. 1H NMR (500 MHz, Chloroform-d): δ 6.66 (s, 1H), 4.56-4.52 (m, 4H), 3.75 (t, J = 1.7 Hz, 2H), 3.52 (q, J = 7.0 Hz, 2H), 3.24-3.18 (m, 2H), 2.85 (t, J = 5.8 Hz, 2H), 2.69-2.61 (m, 2H), 1.21 (t, J = 7.0 Hz, 3H).

[0380] Example 58.5: 2-(Ethoxymethyl)-6-(2-nitroethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine (27.00 mg, 0.0999 mmol), Zn powder (65.30 mg, 0.9987 mmol), and ammonium chloride (53.42 mg, 0.9987 mmol) were suspended in a mixture of THF / water / EtOH (0.9 mL / 0.12 mL / 0.12 mL). The reaction mixture was stirred at room temperature for 1 h 40 min. The mixture was filtered through Celite and rinsed with a 1:1 mixture of DCM:EtOH. The filtrate was passed through a 2 g Biotage NH₂ ion exchange column and rinsed with a 1:1 mixture of DCM:EtOH. The filtrate was concentrated to give a pale yellow oil, 2-[2-(ethoxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridin-6-yl]ethanamine (25 mg, 92%, 0.0915 mmol). HPLC / MS m / z 241.1235 [M+H] + , Rt (AI): 0.30 min. 1 H NMR (600 MHz, Chloroform-d): δ 6.67 (s, 1H), 4.55 (s, 2H), 3.76 (t, J = 1.6 Hz, 2H), 3.52 (q, J = 7.0 Hz, 2H), 2.93 (dd, J = 6.7, 5.1 Hz, 2H), 2.85 (t, J = 5.8 Hz, 2H), 2.75-2.69 (m, 5H), 1.21 (t, J = 7.0 Hz, 3H).

[0381] Example 58.6: 2-[2-(ethoxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridin-6-yl]ethanamine (33.00 mg, 0.1373 mmol) was dissolved in dry DCM (1.06 mL, 0.1300 M), then DIPEA (95.65 μL, 0.5492 mmol) was added, followed by 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (37.43 mg, 0.1647 mmol) and PyBrop (96.00 mg, 0.2059 mmol). The mixture was stirred under argon at room temperature overnight. The volatiles were removed under reduced pressure and the crude product was purified by preparative HPLC (AccqPrep, gradient ACN / water Purification by HPLC (pH 11) afforded N-[2-[2-(ethoxymethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridin-6-yl]ethyl]-7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-amine (5 mg, 8%, 0.0111 mmol) as an off-white solid. HPLC / MS m / z 450.1955 [M+H] + , Rt (AJ): 2.08 min. 1 H NMR (600 MHz, Chloroform-d): δ 8.55-8.53 (m, 1H), 8.21 (dd, J = 8.5, 1.5 Hz, 1H), 8.06 (d, J = 5.8 Hz, 1H), 7.74 (d, J = 8.5 Hz, 1H), 6.94 (dd, J = 5.9, 0.9 Hz, 1H), 6.69 (s, 1H), 6.16 (s, 1H), 4.56 (s, 2H), 3.82-3.75 (m, 4H), 3.53 (q, J = 7.0 Hz, 2H), 2.96 (t, J = 6.0 Hz, 2H), 2.90 (t, J = 5.8 Hz, 2H), 2.76-2.71 (m, 2H), 2.67 (s, 3H), 1.21 (t, J = 7.0 Hz, 3H).

[0382] Example 59: 6-(5-methyl-1,2,4-oxadiazol-3-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one [ka]

[0383] Example 59.1: A solution of 3-(1-chloro-7-isoquinolyl)-5-methyl-1,2,4-oxadiazole [Intermediate A4] (1000.00 mg, 4.0707 mmol) and ethylenediamine (5.44 mL, 81.413 mmol) in NMP (5.00 mL) was placed in a 10-20 mL microwave vial and heated at 160 °C under microwave irradiation for 1 h. Saturated NaHCO solution (100 mL) was added and the product was extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over MgSO and concentrated under reduced pressure to give N'-[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]ethane-1,2-diamine (982 mg, 90%, 3.6465 mmol) as a yellow solid. HPLC / MS m / z 270.1356 [M+H] + , Rt (AJ): 0.95 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.91-8.92 (m, 1H), 8.13 (dd, J = 8.5, 1.6 Hz, 1H), 7.94 (d, J = 5.7 Hz, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.79 (t, J = 5.4 Hz, 1H), 6.92 (dd, J = 5.9, 0.8 Hz, 1H), 3.50 (td, J = 6.4, 5.2 Hz, 2H), 2.81 (t, J = 6.5 Hz, 2H), 2.70 (s, 3H), 1.67 (br s, 2H).

[0384] Example 59.2: A solution of methyl 2-(bromomethyl)-5-cyanobenzoate (215.00 mg, 0.8462 mmol) and N'-[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]ethane-1,2-diamine (250.67 mg, 0.9308 mmol), triethylamine (0.18 mL, 1.2693 mmol) in MeOH (4.23 mL, 0.2000 M) was refluxed under N overnight. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give the crude product, which was washed several times with EtOAc to give 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxo-isoindoline-5-carbonitrile (310.8 mg, 89%, 0.7573 mmol) as a white solid. HPLC / MS m / z 411.1486 [M+H] + , Rt (AI): 1.05 min.

[0385] Example 59.3: 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxo-isoindoline-5-carbonitrile (100.00 mg, 0.2436 mmol), hydroxylamine hydrochloride (33.86 mg, 0.4873 mmol), and TEA (0.07 mL, 0.4873 mmol) were mixed in [bmim]OAc (0.24 mL, 1 M) under argon. The reaction mixture was heated at 80° C. for 1.5 h. The reaction mixture was cooled to ambient temperature and mixed with EtOAc (30 mL). The organic layer was mixed with saturated NaCl (50 mL), and the organic layer was extracted. The aqueous layer was further extracted with EtOAc (2 x 30 mL). The combined organic layers were washed with saturated NaCl (3 x 20 mL) and subsequently concentrated under reduced pressure to afford N-hydroxy-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxo-isoindoline-5-carboxamidine (95 mg, 62%, 0.1500 mmol), which was used in the next step without further purification. HPLC / MS m / z 444.1725 [M+H] + , Rt (AI): 0.76 min.

[0386] Example 59.4: N-hydroxy-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxo-isoindoline-5-carboxamidine (80.00 mg, 0.1263 mmol), acetic anhydride (0.02 mL, 0.2526 mmol), and anhydrous MeCN (1.50 mL, 0.0800 M) were mixed in a microwave vial under argon. The reaction mixture was heated at 160 °C by microwave irradiation for 10 min. Volatiles were removed under reduced pressure. The crude material was directly purified by preparative HPLC (AccqPrep, 30-43% MeOH in water, pH 3, 26 min). The product-containing fractions were filtered through a 1 g SCX2 column. The product was released with 2N ammonia in MeOH to give 6-(5-methyl-1,2,4-oxadiazol-3-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one (23 mg, 39%, 0.0487 mmol) as a white powder. HPLC / MS m / z 468.1787 [M+H] + , Rt (AJ): 2.02 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.81-8.85 (m, 1H), 8.19 (dd, J = 7.9, 1.6 Hz, 1H), 8.15-8.16 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.04 (t, J = 5.6 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.79 (dd, J = 7.8, 0.9 Hz, 1H), 6.95 (dd, J = 5.8, 0.8 Hz, 1H), 4.68 (s, 2H), 3.85-3.90 (m, 2H), 3.78-3.84 (m, 2H), 2.68 (s, 3H), 2.67 (s, 3H).

[0387] Example 60: 6-(3-methyl-1,2,4-oxadiazol-5-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one [ka]

[0388] Example 60.1: tert-Butyl N-[2-(6-bromo-1-oxo-isoindolin-2-yl)ethyl]carbamate (350.00 mg, 0.9853 mmol), acetamide oxime (364.95 mg, 4.9264 mmol), and XantPhos Pd G4 (18.98 mg, 0.0197 mmol) were combined in a microwave vial. The vial was capped, evacuated, and then fitted with a balloon filled with carbon monoxide. Anhydrous 1,4-dioxane (1.97 mL, 0.5000 M) and DIPEA (0.34 mL, 1.9706 mmol) were added, and the reaction mixture was heated at 90 °C for 20 h. Additional XantPhos Pd G4 (18.98 mg, 0.0197 mmol) was added, and the reaction mixture continued to be heated at 90 °C for 5 h. The reaction mixture was evaporated onto silica gel and directly purified by silica gel normal phase flash chromatography (30-100% EtOAc:CycHex) to afford tert-butyl N-[2-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1-oxo-isoindolin-2-yl]ethyl]carbamate (93 mg, 26%, 0.2595 mmol) as a white solid. HPLC / MS m / z 381.1517 [M+Na] + , Rt (AJ): 2.34 min. 1H NMR (600 MHz, DMSO-d6): δ 8.29 (dd, J = 7.9, 1.6 Hz, 1H), 8.23 ​​(d, J = 1.6 Hz, 1H), 7.87 (d, J = 7.9 Hz, 1H), 6.96 (t, J = 6.1 Hz, 1H), 4.63 (s, 2H), 3.59 (t, J = 6.0 Hz, 2H), 3.21 (q, J = 6.1 Hz, 2H), 2.44 (s, 3H), 1.28 (s, 9H).

[0389] Example 60.2: tert-Butyl N-[2-[6-(3-methyl-1,2,4-oxadiazol-5-yl)-1-oxo-isoindolin-2-yl]ethyl]carbamate (90.00 mg, 0.2511 mmol) was mixed with 4 N HCl in 1,4-dioxane (6.28 mL, 25.112 mmol) and anhydrous 1,4-dioxane (6.00 mL) at room temperature under argon and stirred for 3 d. The volatiles were removed under reduced pressure. The crude material was dissolved in MeOH / water and filtered through a 1 g SCX column. The product was released with 2M ammonia in MeOH to give 2-(2-aminoethyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)isoindolin-1-one (64 mg, 99%, 0.2478 mmol) as an off-white solid. HPLC / MS m / z 259.1191 [M+H] + , Rt (AJ): 1.07 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.29 (dd, J = 7.9, 1.6 Hz, 1H), 8.23 ​​(d, J = 1.5 Hz, 1H), 7.85 (dd, J = 7.9, 0.9 Hz, 1H), 4.66 (s, 2H), 3.53 (t, J = 6.4 Hz, 2H), 2.80 (t, J = 6.4 Hz, 2H), 2.44 (s, 3H), 1.69 (s, 2H).

[0390] Example 60.3: 2-(2-aminoethyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)isoindolin-1-one (36.42 mg, 0.1410 mmol), 5-methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (35.24 mg, 0.1551 mmol), PyBroP (72.30 mg, 0.1551 mmol), DIPEA (68.33 mg, 0.5287 mmol), and anhydrous DCM (0.50 mL, 0.2500 M) were placed in a microwave vial under argon at room temperature. The reaction mixture was heated at 60 °C by microwave irradiation for 1 h. The volatiles were removed, and the crude material was directly purified by reverse-phase flash chromatography (30-60% MeOH in water). The product-containing fractions were filtered through a 1 g SCX-2 column. The product was released with 2 M ammonia in MeOH to give 6-(3-methyl-1,2,4-oxadiazol-5-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one (19 mg, 52%, 0.0404 mmol) as an off-white solid. HPLC / MS m / z 468.1786 [M+H] + , Rt (AJ): 2.01 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.82 (s, 1H), 8.28 (dd, J = 7.9, 1.6 Hz, 1H), 8.19-8.21 (m, 1H), 8.14 (dd, J = 8.4, 1.5 Hz, 1H), 8.04 (t, J = 5.5 Hz, 1H), 7.94 (d, J = 5.7 Hz, 1H), 7.83-7.87 (m, 2H), 6.95 (d, J = 5.7 Hz, 1H), 4.71 (s, 2H), 3.86-3.90 (m, 2H), 3.80-3.84 (m, 2H), 2.67 (s, 3H), 2.43 (s, 3H).

[0391] Example 61: Ethyl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate [ka]

[0392] Example 61.1: To a solution of ethyl 1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.300 g, 1.44 mmol) in tetrahydrofuran (5 mL) was added sodium hydride (115 mg, 2.88 mmol, 60% purity) at 0 °C. The mixture was stirred at 0 °C for 0.5 h. Then, 2-bromoacetonitrile (518 mg, 4.32 mmol, 3.00 equiv.) was added to the mixture. The mixture was stirred at 20 °C for 12 h. The mixture was diluted with saturated aqueous ammonium chloride solution (10 mL) at 0 °C and extracted with ethyl acetate (3 × 20 mL). The combined organic phase was washed with brine (3 × 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate - 10 / 1 to 0 / 1) to give ethyl 2-(cyanomethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.130 g, 526 μmol, 36% yield) as a yellow solid. HPLC / MS m / z: 248.2 [M+H] + , Rt (V): 0.79 min. 1 H NMR (400 MHz, DMSO-d6): δ 7.69 (d, J = 1.6 Hz, 1H), 7.02 (d, J = 1.6 Hz, 1H), 4.56 (s, 2H), 4.33-4.28 (m, 2H), 4.21 (q, J = 7.0 Hz, 2H), 3.84-3.77 (m, 2H), 1.27 (t, J = 7.1 Hz, 3H).

[0393] Example 61.2: To a solution of ethyl 2-(cyanomethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.100 g, 404 μmol) in a mixed solvent of methanol (10 mL) and ammonium hydroxide (1 mL) was added Raney nickel (10.0 mg, 10% purity). The mixture was stirred under a hydrogen atmosphere (45 Psi) at 20° C. for 12 h. The mixture was filtered and concentrated under reduced pressure to give ethyl 2-(2-aminoethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (80.0 mg, crude) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6): δ 7.60 (d, J = 1.6 Hz, 1H), 6.88 (d, J = 1.6 Hz, 1H), 4.19 (q, J = 7.0 Hz, 4H), 3.70 (br dd, J = 5.1, 6.7 Hz, 2H), 3.42-3.38 (m, 2H), 2.76-2.65 (m, 2H), 1.25 (t, J = 7.1 Hz, 3H).

[0394] Example 61.3: To a solution of ethyl 2-(2-aminoethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (40.0 mg, 159 μmol) and 7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinoline 2-oxide (36.2 mg, 159 μmol) in dichloromethane (5 mL) was added N,N-diisopropylethylamine (61.7 mg, 478 μmol) and bromotri(pyrrolidin-1-yl)phosphonium hexafluorophosphate (V) (148 mg, 318 μmol). The mixture was stirred at 20° C. for 12 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (ethyl acetate), preparative HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 26%~56%, 8min) and preparative HPLC (column: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225%FA)-ACN]; B%: 18%~38%, 10min) to give ethyl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (10.51mg, 20.54μmol, Yield 12%, purity 99%, formate) was obtained as a white solid. HPLC / MS m / z: 461.2 [M+H] + , Rt (T): 0.78 min. 1H NMR (400 MHz, DMSO-d6): δ 8.87 (s, 1H), 8.34 (s, 1H), 8.14 (dd, J = 1.1, 8.5 Hz, 1H), 8.07-8.01 (m, 1H), 7.96 (d, J = 5.6 Hz, 1H), 7.85 (d, J = 8.5 Hz, 1H), 7.58 (d, J = 1.5 Hz, 1H), 6.96 (d, J = 5.8 Hz, 1H), 6.90 (d, J = 1.6 Hz, 1H), 4.18 (q, J = 6.9 Hz, 4H), 3.81-3.65 (m, 6H), 2.69 (s, 3H), 1.24 (t, J = 7.1 Hz, 3H).

[0395] Example 62: Propan-2-yl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate [ka]

[0396] Example 62.1: To a solution of ethyl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.120 g, 261 μmol) in a mixed solvent of tetrahydrofuran (2 mL) and methanol (2 mL) was added a solution of lithium hydroxide monohydrate (32.8 mg, 782 μmol) in water (1 mL). The mixture was stirred at 70° C. for 12 h. After cooling to room temperature, the mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3μm, mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 4%~34%, 8min) to give 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylic acid (70.0mg, 162μmol, yield 62%) as a yellow oil. HPLC / MS m / z: 433.4 [M+H] + , Rt (V): 0.81 min.

[0397] Example 62.2: To a solution of 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylic acid (50.0 mg, 116 μmol) and potassium carbonate (16.0 mg, 116 μmol) in DMF (2 mL) was added 2-bromopropane (42.7 mg, 347 μmol). The mixture was stirred at 80° C. for 2 h. After cooling to room temperature, the reaction was filtered. The filtrate was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 30%~60%, 8min), preparative HPLC (column: Phenomenex Synergi C18 150*25mm* 10μm; mobile phase: [water (0.225%FA)-ACN]; B%: 12%~42%, 10min) and lyophilized to give propan-2-yl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (18.39mg, 35.0μmol, yield 30%, purity 99%, formate) as a white solid. HPLC / MS m / z: 475.2 [M+H] + , Rt (T): 0.79 min. 1H NMR (400 MHz, DMSO-d6): δ 8.87 (s, 1H), 8.24 (s, 1H), 8.14 (dd, J = 1.5, 8.5 Hz, 1H), 8.07-8.01 (m, 1H), 7.96 (d, J = 5.6 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.54 (d, J = 1.6 Hz, 1H), 6.96 (d, J = 5.8 Hz, 1H), 6.88 (d, J = 1.8 Hz, 1H), 5.01 (sept, J = 6.3 Hz, 1H), 4.22-4.12 (m, 2H), 3.76-3.69 (m, 6H), 2.69 (s, 3H), 1.24 (d, J = 6.3 Hz, 6H).

[0398] Example 63: Ethyl 5-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate [ka]

[0399] Example 63.1: To a mixture of 2-bromo-6,7-dihydro-5H-thieno[3,2-c]pyridin-4-one (800 mg, 3.45 mmol) in ethanol (20 mL), palladium acetate (77.4 mg, 34.5 μmol), triethylamine (69.8 mg, 6.89 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (598 mg, 1.03 mmol), and DMF (4 mL) were added. The mixture was stirred at 70° C. under a carbon monoxide atmosphere (40 Psi) for 12 h. After the reaction was complete, the mixture was cooled to 25° C. and quenched with water (5 mL). The suspension was filtered, and the filtrate was concentrated to remove ethanol. The residue was extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (3x20 mL), dried over anhydrous sodium sulfate, and concentrated to give a residue, which was purified by column chromatography (SiO2, Purification by petroleum ether / ethyl acetate - 5 / 1 to 1 / 1) gave ethyl 4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate (750 mg, 3.33 mmol, yield 97%) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6): δ 7.79 (s, 2H), 4.28 (q, J = 7.2 Hz, 2H), 3.47 (dt, J = 2.8, 6.8 Hz, 2H), 3.06 (t, J = 6.8 Hz, 2H), 1.29 (t, J = 7.2Hz, 3H).

[0400] Example 63.2: To a suspension of sodium hydride (24.9 mg, 6.21 mmol, purity 60%) in tetrahydrofuran (7 mL) was added ethyl 4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate (700 mg, 3.11 mmol). The mixture was stirred at 0° C. for 0.5 h. Then, 2-bromoacetonitrile (1.12 g, 9.32 mmol) was added to the mixture. The mixture was stirred at 25° C. for 2 h. The mixture was quenched with saturated aqueous ammonium chloride solution (5 mL). The mixture was concentrated to give a residue. The residue was purified by column chromatography (SiO 2, Purification by petroleum ether / ethyl acetate - 5 / 1 to 1 / 1) gave ethyl 5-(cyanomethyl)-4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate (450 mg, 1.70 mmol, yield 54%) as a yellow solid. HPLC / MS m / z: 265.1 [M+H] + , Rt (T): 0.86 min. 1 H NMR (400 MHz, Chloroform-d): δ 8.08 (s, 1H), 4.51 (s, 2H), 4.36 (q, J = 7.2 Hz, 2H), 3.82 (t, J = 6.8 Hz, 2H), 3.24 (t, J = 6.8 Hz, 2H), 1.38 (t, J = 7.2 Hz, 3H).

[0401] Example 63.3: A mixture of ethyl 5-(cyanomethyl)-4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate (250 mg, 94.6 μmol), Raney nickel (81.0 mg, 94.6 μmol, 10% purity), and ammonium hydroxide (1 mL) in ethanol (10 mL) was stirred under a hydrogen atmosphere (45 Psi) at 25° C. for 12 h. After the reaction was complete, the mixture was filtered and the filtrate was concentrated to give a residue. The residue was purified by preparative TLC (ethyl acetate / methanol / ammonium hydroxide (25% purity) - 50 / 10 / 1) to afford ethyl 5-(2-aminoethyl)-4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate (75.0 mg, 280 μmol, yield 29%) as a colorless solid. 1 H NMR (400 MHz, Chloroform-d): δ 8.00 (s, 1H), 4.34 (q, J = 7.2 Hz, 2H), 3.75 (t, J = 6.8 Hz, 2H), 3.71 (br t, = 6.0 Hz, 2H), 3.17-3.10 (m, 4H), 1.37 (t, J = 7.2 Hz, 3H).

[0402] Example 63.4: A mixture of 7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinoline 2-oxide (50.0 mg, 220 μmol), ethyl 5-(2-aminoethyl)-4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate (59.0 mg, 220 μmol), diisopropylethylamine (11.4 mg, 880 μmol), and bromotri(pyrrolidin-1-yl)phosphonium hexafluorophosphate (V) (18.5 mg, 396 μmol) in dichloromethane (5 mL) was stirred for 12 h at 25° C. The mixture was concentrated to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge 150*25mm* 5μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 46%~76%, 10min), preparative HPLC (column: Phenomenex Synergi C18 150*25mm* 10μm; mobile phase: [water (0.225%FA)-ACN]; B%: 20%~47%, 9min), and lyophilized to give ethyl 5-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-4-oxo-4,5,6,7-tetrahydrothieno[3,2-c]pyridine-2-carboxylate (16.99mg, 32.1μmol, yield 14%, purity 99%, formate) as a colorless solid. HPLC / MS m / z: 478.2 [M+H] + , Rt (T): 0.84 min. 1 H NMR 1H NMR (400 MHz, DMSO-d6): δ 8.87 (s, 1H), 8.20-8.11 (m, 1H), 8.02 (br s, 1H), 7.96 (d, J = 5.6 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.79 (s, 1H), 6.95 (d, J = 5.6 Hz, 1H), 4.28 (q, J = 7.2 Hz, 2H), 3.76-3.66 (m, 6H), 3.09 (t, J = 6.8 Hz, 2H), 2.69 (s, 3H), 1.28 (t, J = 7.2Hz, 3H).

[0403] Example 64: 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-7-(5-methyl-1,3,4-oxadiazol-2-yl)-3,4-dihydropyrrolo[1,2-a]pyrazin-1(2H)-one [ka] A mixture of 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylic acid (40.0 mg, 92.5 μmol) [Example 62.1] and acetohydrazide (8.91 mg, 120 μmol) in polyphosphoric acid (0.5 mL) was stirred at 90° C. for 2 h. The reaction was diluted with ethyl acetate (10 mL). The pH of the mixture was then adjusted to 8 with saturated aqueous sodium carbonate solution. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Waters Xbridge 150*25 mm* 5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 20%~50%, 9 min) and lyophilized to give 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-7-(5-methyl-1,3,4-oxadiazol-2-yl)-3,4-dihydropyrrolo[1,2-a]pyrazin-1(2H)-one (10.78 mg, 22.7 μmol, yield 24%, purity 99%) as a colorless solid. HPLC / MS m / z: 471.3 [M+H] + , Rt (V): 0.90 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.92 (br s, 1H), 8.21 (br s, 1H), 7.94 (br d, J = 5.7 Hz, 1H), 7.93-7.84 (m, 1H), 7.68 (d, J = 1.7 Hz, 1H), 7.02 (br d, J = 2.1 Hz, 1H), 6.97 (d, J = 1.6 Hz, 1H), 4.23 (br t, J = 5.6 Hz, 2H), 3.77 (br s, 6H), 2.69 (s, 3H), 2.50 (s, 3H).

[0404] Example 65: 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-7-(3-methyl-1,2,4-oxadiazol-5-yl)-3,4-dihydropyrrolo[1,2-a]pyrazin-1(2H)-one [ka]

[0405] Example 65.1: To a solution of 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylic acid (80.0 mg, 185 μmol) [Example 62.1], 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (70.9 mg, 370 μmol), 1-hydroxybenzotriazole (37.5 mg, 27.8 μmol), and N,N-diisopropylethylamine (71.7 mg, 555 μmol, 96.7 μL) in acetonitrile (5 mL) was added N-hydroxyacetamidine (27.4 mg, 370 μmol). The mixture was stirred at 20° C. for 12 h. The mixture was concentrated to give a residue. The residue was triturated with water (3 mL) to give N'-((2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carbonyl)oxy)acetimidamide (90.0 mg, crude) as a yellow solid. HPLC / MS m / z: 489.3 [M+H] + , Rt (V): 0.89 min. 1H NMR (400 MHz, DMSO-d6): δ 8.88 (s, 1H), 8.20-8.11 (m, 1H), 8.06 (br s, 1H), 7.97 (d, J = 5.8 Hz, 1H), 7.85 (d, J = 8.5 Hz, 1H), 7.75 (d, J = 1.3 Hz, 1H), 7.16 (d, J = 1.4 Hz, 1H), 6.96 (d, J = 5.6 Hz, 1H), 6.38 (br s, 3H), 4.16 (br t, J = 5.4 Hz, 2H), 3.74 (br s, 6H), 2.69 (s, 3H), 2.50 (br s, 3H).

[0406] Example 65.2: To a solution of N'-((2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carbonyl)oxy)acetimidamide (60.0 mg, 12.3 μmol) in tetrahydrofuran (3 mL) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran, 368 μL, 368 μmol). The mixture was stirred at 40° C. for 12 h. The mixture was diluted with saturated aqueous ammonium chloride solution (10 mL) and extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (3×10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (ethyl acetate), preparative HPLC (column: Waters Xbridge 150*25 mm*5 μm, mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 29%~59%, 8 min), and lyophilized to give 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-7-(3-methyl-1,2,4-oxadiazol-5-yl)-3,4-dihydropyrrolo[1,2-a]pyrazin-1(2H)-one (13.02 mg, 27.40 μmol, yield 22%, purity 99%) as a colorless solid. HPLC / MS m / z: 471.3 [M+H]+, Rt (T): 0.77 min. 1H NMR (400 MHz, DMSO-d6): δ 8.87 (s, 1H), 8.15 (dd, J = 1.3, 8.6 Hz, 1H), 8.09-8.02 (m, 1H), 7.97 (d, J = 5.7 Hz, 1H), 7.88-7.80 (m, 2H), 7.06 (d, J = 1.7 Hz, 1H), 6.96 (d, J = 5.7 Hz, 1H), 4.24 (br t, J = 5.7 Hz, 2H), 3.76 (br s, 6H), 2.68 (s, 3H), 2.33 (s, 3H).

[0407] Example 66: Ethyl 1-methyl-5-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carboxylate [ka]

[0408] Example 66.1: To a mixture of DMF (11.4 g, 15.6 mmol, 1.2 mL) and dichloromethane (40 mL) was added phosphorus oxychloride (13.2 g, 86.3 mmol) slowly at 0° C. The mixture was stirred at 0° C. for 15 min. To the mixture was added a solution of benzyl 4-oxopiperidine-1-carboxylate (10.0 g, 42.9 mmol) in dichloromethane (40 mL) slowly. The resulting mixture was stirred at 25° C. for 2 h. After the reaction was completed, the reaction mixture was poured into a solution of sodium acetate (10.0 g) in ice water (100 mL). The reaction mixture was extracted with dichloromethane (3×40 mL), washed with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate - 20 / 1 to 8 / 1) to afford benzyl 4-chloro-3-formyl-5,6-dihydropyridine-1(2H)-carboxylate (5.40 g, 19.3 mmol, yield 45%) as a pale yellow oil. 1 H NMR (400 MHz, Chloroform-d): δ 10.14 (s, 1H), 7.40-7.33 (m, 5H), 5.16 (s, 2H), 4.23 (br s, 2H), 3.70 (t, J = 5.6 Hz, 2H), 2.70 (br s, 2H).

[0409] Example 66.2: To a solution of benzyl 4-chloro-3-formyl-5,6-dihydropyridine-1(2H)-carboxylate (5.40 g, 19.3 mmol) in dichloromethane (100 mL) was added ethyl 2-(triphenylphosphoranylidene)acetate (6.73 g, 19.3 mmol). The mixture was stirred at 25° C. for 12 h. After the reaction was complete, the mixture was concentrated to give a residue. The residue was purified by column chromatography (SiO 2, Purification by petroleum ether / ethyl acetate - 50 / 1 to 5 / 1) gave benzyl 4-chloro-3-(3-ethoxy-3-oxoprop-1-en-1-yl)-5,6-dihydropyridine-1(2H)-carboxylate (5.92 g, 16.9 mmol, 87% yield) as a yellow oil. HPLC / MS m / z: 350.1 [M+H] + , Rt (T): 1.02 min. 1 H NMR (400 MHz, Chloroform-d): δ 7.84 (d, J = 16.4 Hz, 1H), 7.41-7.35 (m, 5H), 6.06-5.78 (m, 1H), 5.19-5.15 (m, 2H), 4.25 (q, J = 7.2 Hz, 4H), 3.69 (t, J = 6.0 Hz, 2H), 2.76-2.53 (m, 2H), 1.32 (t, J = 7.2 Hz, 3H).

[0410] Example 66.3: A mixture of benzyl 4-chloro-3-(3-ethoxy-3-oxoprop-1-en-1-yl)-5,6-dihydropyridine-1(2H)-carboxylate (3.90 g, 11.15 mmol) and sodium azide (1.96 g, 30.15 mmol, 2.70 equiv) in DMF (40 mL) was stirred at 25° C. for 8 h. The mixture was poured into water (40 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (3 x 20 mL), dried over anhydrous sodium sulfate and concentrated to afford benzyl 4-azido-3-(3-ethoxy-3-oxoprop-1-en-1-yl)-5,6-dihydropyridine-1(2H)-carboxylate (4.6 g, crude) as a yellow oil, which was used directly in the next step. HPLC / MS m / z: 379.1 [M+Na], Rt (T): 1.09 min.

[0411] Example 66.4: A solution of benzyl 4-azido-3-(3-ethoxy-3-oxoprop-1-en-1-yl)-5,6-dihydropyridine-1(2H)-carboxylate (4.60 g, 12.9 mmol) in toluene (45 mL) was stirred at 60 °C for 8 h. After the reaction was completed, the mixture was concentrated to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate - 10 / 1 to 3 / 1) to give 5-benzyl 2-ethyl 6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-2,5(4H)-dicarboxylate (700 mg, crude) as a yellow oil. 1 H NMR (400 MHz, Chloroform-d): δ 8.86 (br s, 1H), 7.45-7.30 (m, 5H), 6.67 (s, 1H), 5.17 (s, 2H), 4.50 (s, 2H), 4.30 (q, J = 7.2 Hz, 2H), 3.85-3.75 (m, 2H), 2.80-2.68 (m, 2H), 1.35 (t, J = 7.2 Hz, 3H).

[0412] Example 66.5: To a suspension of cesium carbonate (1.39 g, 4.26 mmol) in acetonitrile (10 mL) was added 5-benzyl 2-ethyl 6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-2,5(4H)-dicarboxylate (700 mg, 2.13 mmol) and iodomethane (1.56 g, 11.0 mmol, 5.16 equiv.). The mixture was stirred at 50 °C for 12 h. After the reaction was complete, the mixture was poured into water (5 mL). The mixture was extracted with ethyl acetate (3 × 5 mL), and the combined organic layers were concentrated to give 5-benzyl 2-ethyl 1-methyl-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-2,5(4H)-dicarboxylate (690 mg, crude) as a yellow oil. HPLC / MS m / z: 343.2 [M+H] + , Rt (U): 1.12 min. 1 H NMR (400 MHz, Chloroform-d): δ 7.39-7.34 (m, 5H), 6.75 (br s, 1H), 5.16 (s, 2H), 4.48 (s, 2H), 4.27 (q, J = 7.2 Hz, 2H), 3.86-3.77 (m, 5H), 2.67 (br s, 2H), 1.34 (t, J = 7.2 Hz, 3H).

[0413] Example 66.6: A mixture of 5-benzyl 2-ethyl 1-methyl-6,7-dihydro-1H-pyrrolo[3,2-c]pyridine-2,5(4H)-dicarboxylate (0.7 g, 2.04 mmol) and palladium on activated carbon (0.100 g, 5% purity) in methanol (20 mL) was stirred under a hydrogen atmosphere (15 Psi) at 25° C. for 12 h. After the reaction was complete, the mixture was filtered. The filtrate was concentrated to give ethyl 1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carboxylate (0.22 g, crude) as a colorless oil. 1H NMR (400 MHz, Chloroform-d): δ 6.76 (s, 1H), 4.30-4.25 (m, 2H), 4.11 (s, 2H), 3.80-3.78 (m, 3H), 3.44 (br t, J = 6.0 Hz, 2H), 2.94 (br t, J = 5.6 Hz, 2H), 1.34 (t, J = 7.2 Hz, 3H).

[0414] Example 66.7: A mixture of ethyl 1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carboxylate (200 mg, 960 μmol), tert-butyl N-((2-bromoethyl)carbamate (258 mg, 1.15 mmol), sodium iodide (288 mg, 1.92 mmol), and cesium carbonate (626 mg, 1.92 mmol) in acetonitrile (0.5 mL) was stirred at 60° C. for 2 h. After the reaction was complete, the mixture was filtered. The filtrate was concentrated to give a residue. The residue was purified by column chromatography (SiO 2, Purification with petroleum ether / ethyl acetate - 8 / 1 to 3 / 1) afforded ethyl 5-(2-((tert-butoxycarbonyl)amino)ethyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carboxylate (260 mg, 739 μmol, yield 77%) as a colorless oil. 1 H NMR (400 MHz, Chloroform-d): δ 6.76 (s, 1H), 4.30-4.25 (m, 2H), 3.92 (s, 1H), 3.81 (s, 3H), 3.78-3.70 (m, 1H), 3.68-3.57 (m, 2H), 3.50 (s, 1H), 3.49-3.27 (m, 2H), 3.31-3.23 (m, 1H), 3.16-3.11 (m, 1H), 3.05-2.91 (m, 2H), 1.45 (s, 9H), 1.36-1.32 (m, 3H).

[0415] Example 66.8: A mixture of ethyl 5-(2-((tert-butoxycarbonyl)amino)ethyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carboxylate (50.0 mg, 142 μmol) in hydrochloric acid / ethyl acetate (4 M, 0.5 mL) was stirred at 25° C. for 1 h. The mixture was concentrated to give ethyl 5-(2-aminoethyl)-1-methyl-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carboxylate (41.0 mg, crude, hydrochloride salt) as a gray solid. HPLC / MS m / z: 252.5 [M+H] + , Rt (T): 0.22 min.

[0416] Example 66.9: A mixture of ethyl 5-(2-aminoethyl)-1-methyl-6,7-dihydro-4H-pyrrolo[3,2-c]pyridine-2-carboxylate (41.0 mg, 142 μmol, hydrochloride), diisopropylethylamine (73.6 mg, 5.70 μmol), 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (32.4 mg, 142 μmol), and bromo(tripyrrolidin-1-yl)phosphonium hexafluorophosphate (1.20 mg, 256 μmol) in DMF (2.50 mL) was stirred for 12 h at 25° C. After the reaction was complete, the pH of the mixture was adjusted to approximately 7 by adding hydrochloric acid (1 M). The mixture was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 44%~74%, 11.5min) and lyophilized to give ethyl 1-methyl-5-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine-2-carboxylate (17.89mg, 38.8μmol, yield 27%) as a yellow solid. HPLC / MS m / z: 461.2 [M+H] +, Rt (V): 1.03 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.86 (s, 1H), 8.13 (dd, J = 1.2, 8.4 Hz, 1H), 7.96 (d, J = 5.6 Hz, 1H), 7.88-7.78 (m, 2H), 6.93 (d, J = 5.6 Hz, 1H), 6.62 (s, 1H), 4.16 (q, J = 7.2 Hz, 2H), 3.73-3.65 (m, 5H), 3.44 (s, 2H), 2.85-2.75 (m, 4H), 2.70 (s, 3H), 2.67-2.61 (m, 2H), 1.24 (t, J = 7.2 Hz, 3H).

[0417] Example 67: Ethyl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate [ka]

[0418] Example 67.1: To a solution of 7-bromo-3,4-dihydroisoquinolin-1(2H)-one (1.50 g, 6.64 mmol), triethylamine (1.34 g, 13.3 mmol) in a mixed solvent of ethanol (20 mL) and DMF (4 mL) was added palladium(II) acetate (14.9 mg, 66.4 μmol) and (5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)diphenyl-phosphane (1.15 g, 1.99 mmol). The mixture was stirred at 70° C. under a carbon monoxide atmosphere (45 Psi) for 24 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give ethyl 1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate (0.900 g, 4.11 mmol, yield 61%) as a yellow solid. HPLC / MS m / z: 220.0 [M+H]+ , Rt (V): 0.81 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.41 (d, J = 1.6 Hz, 1H), 8.11 (br s, 1H), 8.03 (dd, J = 1.7, 7.8 Hz, 1H), 7.47 (d, J = 7.9 Hz, 1H), 4.33 (q, J = 7.1 Hz, 2H), 3.39 (td, J = 3.2, 6.5 Hz, 2H), 2.99 (t, J = 6.5 Hz, 2H), 1.33 (t, J = 7.1 Hz, 3H).

[0419] Example 67.2: To a solution of ethyl 1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate (0.800 g, 3.65 mmol) in tetrahydrofuran (10 mL) was added sodium hydride (204 mg, 5.11 mmol, 60% purity) at 0 °C. The mixture was stirred at 0 °C for 0.5 h. Then, 2-bromoacetonitrile (1.31 g, 11.0 mmol) was added to the mixture. The mixture was stirred at 20 °C for 12 h. The mixture was diluted with saturated aqueous ammonium chloride solution (20 mL) at 0 °C and extracted with ethyl acetate (3 × 20 mL). The combined organic layers were washed with brine (3 × 20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate - 10 / 1 to 2 / 1) to give ethyl 2-(cyanomethyl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate (0.600 g, 2.32 mmol, yield 63%) as a yellow solid. 1H NMR (400 MHz, DMSO-d6): δ 8.46 (d, J = 1.5 Hz, 1H), 8.09 (dd, J = 1.7, 7.9 Hz, 1H), 7.51 (d, J = 8.1 Hz, 1H), 4.63 (s, 2H), 4.34 (q, J = 7.0 Hz, 2H), 3.71 (t, J = 6.6 Hz, 2H), 3.18-3.10 (m, 2H), 1.34 (t, J = 7.1 Hz, 3H).

[0420] Example 67.3: To a solution of ethyl 2-(cyanomethyl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate (600 mg, 2.32 mmol) in a mixed solvent of methanol (20 mL) and ammonium hydroxide (4 mL) was added Raney nickel (60.0 mg, 10% purity). The mixture was stirred under a hydrogen atmosphere (45 Psi) at 20° C. for 12 h. The mixture was filtered and concentrated to give a residue. The residue was purified by reverse-phase column chromatography (C18, 40 g, condition: water / acetonitrile - 1 / 0~0 / 1, 0.1% formic acid) and preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 8%~38%, 8 min) to give ethyl 2-(2-aminoethyl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate (0.250 g, 953 μmol, yield 41%) as a yellow oil. 1 H NMR (400 MHz, Methanol-d4) δ 8.64 (d, J = 1.6 Hz, 1H), 8.15 (dd, J = 2.0, 7.9 Hz, 1H), 7.46 (d, J = 8.1 Hz, 1H), 4.41 (q, J = 7.2 Hz, 2H), 3.87 (t, J = 5.9 Hz, 2H), 3.73 (t, J = 6.7 Hz, 2H), 3.26 (t, J = 5.9 Hz, 2H), 3.18 (t, J = 6.6 Hz, 2H), 1.42 (t, J = 7.2 Hz, 3H).

[0421] Example 67.4: To a solution of ethyl 2-(2-aminoethyl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate (60.0 mg, 201 μmol, hydrochloride salt) in dichloromethane (5 mL), N,N-diisopropylethylamine (123 mg, 953 μmol), 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (43.3 mg, 19.1 μmol) and bromotripyrrolidinophosphonium hexafluorophosphate (17.8 mg, 381 μmol) were added. The mixture was stirred at 25° C. for 12 h. The mixture was concentrated under reduced pressure. The residue was purified by preparative TLC (petroleum ether / ethyl acetate - 1 / 1), preparative HPLC (column: Phenomenex Gemini-NX C18 75*30 mm*3 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 36%~66%, 8 min) and lyophilized to give ethyl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1-oxo-1,2,3,4-tetrahydroisoquinoline-7-carboxylate (10.94 mg, 22.97 μmol, yield 12%, purity 99%) as a yellow solid. HPLC / MS m / z: 472.2 [M+H] + , Rt (V): 1.01 min. 1H NMR (400 MHz, DMSO-d6): δ8.87 (s, 1H), 8.40 (d, J = 1.7 Hz, 1H), 8.15 (dd, J = 1.4, 8.5 Hz, 1H), 8.06 (br t, J = 5.1 Hz, 1H), 8.01 (dd, J = 1.9, 7.9 Hz, 1H), 7.96 (d, J = 5.7 Hz, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.43 (d, J = 7.8 Hz, 1H), 6.95 (d, J = 5.7 Hz, 1H), 4.32 (q, J = 7.1 Hz, 2H), 3.83-3.73 (m, 4H), 3.62 (t, J = 6.5 Hz, 2H), 3.01 (t, J = 6.5 Hz, 2H), 2.68 (s, 3H), 1.32 (t, J = 7.1 Hz, 3H).

[0422] Example 68: Propan-2-yl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate [ka]

[0423] Example 68.1: To a solution of ethyl 2-oxopropanoate (20.0 g, 172 mmol) in acetonitrile (150 mL) was added ethane-1,2-diamine (10.4 g, 172 mmol). The mixture was stirred at 20° C. for 0.5 h. To the mixture was added a solution of ethyl 3-bromo-2-oxopropanoate (33.6 g, 172 mmol) and iron(III) chloride (5.59 g, 34.5 mmol) in acetonitrile (60 mL) at 0° C. The mixture was stirred at 80° C. for 12 h. After cooling to room temperature, the mixture was diluted with saturated aqueous ammonium chloride solution (150 mL) and extracted with ethyl acetate (3×150 mL). The combined organic layers were washed with brine (3×100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO 2, Purification by petroleum ether / ethyl acetate - 5 / 1 to ethyl acetate / methanol - 10 / 1) gave ethyl 1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (4.00 g, 19.2 mmol, yield 11%) as a brown solid. 1 H NMR (400 MHz, DMSO-d6): δ 7.92 (br s, 1H), 7.64 (s, 1H), 6.91 (s, 1H), 4.25-4.18 (m, 2H), 4.18-4.12 (m, 2H), 3.51 (br s, 2H), 1.26 (br t, J = 7.0 Hz, 3H).

[0424] Example 68.2: To a solution of ethyl 1-oxo-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (4.00 g, 19.2 mmol) and boron trifluoride diethyl etherate (46.0 g, 324 mmol, 40.0 mL) in tetrahydrofuran (300 mL) was added sodium borohydride (8.00 g, 211 mmol) portionwise at 0° C. The mixture was stirred at 20° C. for 24 h. The mixture was quenched with ethyl alcohol (40 mL) and stirred for 12 h. The mixture was concentrated to give a residue. The residue was diluted with aqueous hydrochloric acid (10%, 30 mL), and the solution was stirred for 2 h. The resulting solution was neutralized with saturated aqueous sodium bicarbonate solution until the pH of the solution was 8. The mixture was diluted with ethyl acetate (200 mL). After separation, the aqueous layer was extracted with ethyl acetate (2 × 200 mL). The combined organic layers were concentrated to give a residue. The residue was purified by reverse-phase column chromatography (C18, 40 g, conditions: water / acetonitrile - 1 / 0 to 0 / 1, 0.1% trifluoroacetic acid) to give ethyl 1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (1.00 g, 3.24 mmol, yield 16%, trifluoroacetate) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6): δ 9.36 (br s, 2H), 7.48 (d, J = 1.6 Hz, 1H), 6.36 (d, J = 0.8 Hz, 1H), 4.31 (s, 2H), 4.24-4.18 (m, 2H), 4.18-4.12 (m, 2H), 3.56 (br t, J = 5.8 Hz, 2H), 1.23 (t, J = 7.1 Hz, 3H).

[0425] Example 68.3: To a suspension of ethyl 1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.400 g, 1.30 mmol, trifluoroacetate), tert-butyl N-(2-bromoethyl)carbamate (1.15 g, 5.13 mmol) in DMF (5 mL) was added N,N-diisopropylethylamine (798 mg, 6.18 mmol) and potassium iodide (34.2 mg, 206 μmol). The mixture was stirred at 90° C. for 12 h. The mixture was concentrated to give a residue. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate - 10 / 1 to 0 / 1) to afford ethyl 2-(2-((tert-butoxycarbonyl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.350 g, 1.04 mmol, yield 80%) as a yellow oil. HPLC / MS m / z: 338.1 [M+H] + , Rt (V): 0.92 min. 1 H NMR (400 MHz, DMSO-d6): δ 7.28 (d, J = 1.6 Hz, 1H), 6.73 (br t, J = 5.4 Hz, 1H), 6.10 (s, 1H), 4.13 (q, J = 7.1 Hz, 2H), 3.94 (br t, J = 5.3 Hz, 2H), 3.54 (s, 2H), 3.13-3.05 (m, 2H), 2.77 (br t, J = 5.4 Hz, 2H), 2.47 (br s, 2H), 1.24-1.20 (m, 3H).

[0426] Example 68.4: A solution of ethyl 2-(2-((tert-butoxycarbonyl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.350 g, 1.04 mmol) in hydrochloric acid solution / ethyl acetate (4 M, 5 mL) was stirred at 20° C. for 2 h. The mixture was concentrated under reduced pressure to afford ethyl 2-(2-aminoethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.25 g, crude, hydrochloride salt) as a yellow solid. 1 H NMR (400 MHz, Methanol-d4): δ 7.43 (d, J = 1.6 Hz, 1H), 6.46 (s, 1H), 4.44 (s, 2H), 4.38 (t, J = 5.8 Hz, 2H), 4.24 (q, J = 7.1 Hz, 2H), 3.74-3.69 (m, 2H), 3.56-3.42 (m, 4H), 1.31 (t, J = 7.2 Hz, 3H).

[0427] Example 68.5: To a solution of ethyl 2-(2-aminoethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (0.150 g, 548 μmol, hydrochloride), 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (125 mg, 548 μmol), and N,N-diisopropylethylamine (354 mg, 2.74 mmol) in dichloromethane (2 mL) was added bromotripyrrolidinophosphonium hexafluorophosphate (511 mg, 1.10 mmol). The mixture was stirred at 35° C. for 12 h. The mixture was concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (ethyl acetate) and reverse-phase column chromatography (C18, 40 g; conditions: water / acetonitrile - 1 / 0 to 0 / 1, 0.1% ammonium hydroxide) to give ethyl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (50.0 mg, 112 μmol, yield 20%) as a yellow oil. 1 H NMR (400 MHz, DMSO-d6): δ 8.87 (s, 1H), 8.14 (dd, J = 1.5, 8.4 Hz, 1H), 7.96 (d, J = 5.7 Hz, 1H), 7.90-7.78 (m, 2H), 7.29 (d, J = 1.7 Hz, 1H), 6.94 (d, J = 5.6 Hz, 1H), 6.12 (d, J = 1.5 Hz, 1H), 4.13 (q, J = 7.1 Hz, 2H), 3.97 (t, J = 5.5 Hz, 2H), 3.70 (q, J = 6.2 Hz, 2H), 3.65 (s, 2H), 2.88 (t, J = 5.5 Hz, 2H), 2.79 (t, J = 6.7 Hz, 2H), 2.70 (s, 3H), 1.22 (t, J = 7.1 Hz, 3H).

[0428] Example 68.6: To a solution of ethyl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (45.0 mg, 101 μmol) in a mixed solvent of tetrahydrofuran (2 mL) and methanol (2 mL) was added a solution of lithium hydroxide monohydrate (12.7 mg, 302 μmol) in water (1 mL). The mixture was stirred at 70° C. for 12 h. The mixture was then concentrated under reduced pressure to afford 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylic acid (50 mg, crude, lithium salt) as a yellow solid. 1 H NMR (400 MHz, DMSO-d6): δ 8.88 (s, 1H), 8.15-8.13 (m, 1H), 7.97 (d, J = 5.6 Hz, 1H), 7.87-7.82 (m, 1H), 6.94 (d, J = 5.7 Hz, 1H), 6.69 (s, 1H), 5.84 (s, 1H), 3.89-3.83 (m, 2H), 3.71 (m, 2H), 3.02 (dt, J = 3.7, 6.4 Hz, 2H), 2.82 (br d, J = 4.4 Hz, 2H), 2.77 (br d, J = 6.5 Hz, 2H), 2.70 (s, 3H).

[0429] Example 68.7: To a solution of 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylic acid (50 mg, crude, lithium salt) in DMF (2.5 mL) was added a solution of 2-bromopropane (18.8 mg, 153 μmol) in DMF (0.5 mL). The mixture was stirred at 50° C. for 4 h. Then the mixture was filtered. The filtrate was purified by preparative HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (10mM NH4HCO3)-ACN]; B%: 40%~70%, 8min) and lyophilized to give propan-2-yl 2-(2-((7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-1,2,3,4-tetrahydropyrrolo[1,2-a]pyrazine-7-carboxylate (15.64mg, 33.6μmol, yield 28%, purity 99%) as a yellow solid. HPLC / MS m / z: 461.2 [M+H] + , Rt (V): 1.03 min. 1 H NMR (400 MHz, DMSO-d6): δ 8.87 (s, 1H), 8.14 (dd, J = 1.5, 8.6 Hz, 1H), 7.96 (d, J = 5.7 Hz, 1H), 7.89-7.85 (m, 1H), 7.84 (d, J = 8.6 Hz, 1H), 7.25 (d, J = 1.7 Hz, 1H), 6.94 (d, J = 5.6 Hz, 1H), 6.10 (d, J = 1.6 Hz, 1H), 4.98 (quin, J = 6.2 Hz, 1H), 3.97 (t, J = 5.4 Hz, 2H), 3.70 (q, J = 6.3 Hz, 2H), 3.65 (s, 2H), 2.88 (br t, J = 5.5 Hz, 2H), 2.78 (t, J = 6.7 Hz, 2H), 2.70 (s, 3H), 1.22 (d, J = 6.2 Hz, 6H).

[0430] Example 69: Ethyl 6-[2-[[3-(5-methyl-1,2,4-oxadiazol-3-yl)benzoyl]amino]ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate [ka]

[0431] Example 69.1: 6-(tert-Butoxycarbonyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-carboxylic acid (100.00 mg, 0.3529 mmol), 4-dimethylaminopyridine (12.94 mg, 0.1059 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (101.49 mg, 0.5294 mmol) were dissolved in a mixture of DMF (1.01 mL) and EtOH (1.00 mL). The reaction mixture was stirred at 70 °C for 45 min. The mixture was cooled to room temperature, water and ethyl acetate were added, and the product was extracted with EtOAc, dried over MgSO4, and concentrated onto silica in vacuo. The product was purified using an Isco Combi-flash purification system (eluent cyclohexane / EtOAc 20% to 80%, 8 min) to give O6-tert-butyl O2-ethyl 5,7-dihydro-4H-thieno[2,3-c]pyridine-2,6-dicarboxylate (80 mg, 73%, 0.2569 mmol) as a colorless oil. HPLC / MS m / z: 212.07 [M+H-Boc]+, Rt (AE): 1.54 min.

[0432] Example 69.2: O6-tert-Butyl O2-ethyl 5,7-dihydro-4H-thieno[2,3-c]pyridine-2,6-dicarboxylate (70.00 mg, 0.2248 mmol) was dissolved in dry 1,4-dioxane (1.12 mL). Hydrogen chloride (4 M in 1,4-dioxane) (0.56 mL, 2.2479 mmol) was added, and the reaction mixture was stirred at room temperature for 18 h. Additional hydrogen chloride (4 M in 1,4-dioxane) (0.56 mL, 2.2479 mmol) was added, and the reaction mixture was stirred for an additional 6 h. The solvent was removed in vacuo to give ethyl 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-carboxylate hydrochloride (48 mg, 86%, 0.1938 mmol) as a colorless powder. HPLC / MS m / z: 212.07 [M+H]+, Rt (AE): 0.69 min.

[0433] Example 69.3: Ethyl 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-carboxylate hydrochloride (48.00 mg, 0.1938 mmol) and N-Boc-2-chloroethylamine (88.79 mg, 0.4844 mmol) were dissolved in dry DMF (0.97 mL). Triethylamine (81.83 μL, 0.5813 mmol) was added, and the reaction mixture was stirred at 70 °C for 18 h. Water and EtOAc were added. The product was extracted with EtOAc, dried over MgSO and concentrated in vacuo. Purification by silica gel column chromatography (eluent: 20-60% EtOAc in cyclohexane) afforded ethyl 6-[2-(tert-butoxycarbonylamino)ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate (26 mg, 38%, 0.0734 mmol) as an orange oil. HPLC / MS m / z: 355.107 [M+H] + , Rt (AE): 1.03 min.

[0434] Example 69.4: Ethyl 6-[2-(tert-butoxycarbonylamino)ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate (26.00 mg, 0.0734 mmol) was dissolved in dry 1,4-dioxane (0.37 mL). Hydrogen chloride (4 M in dioxane) (0.18 mL, 0.7335 mmol) was added, and the reaction mixture was stirred at room temperature for 2 h. The solvent was removed in vacuo to give ethyl 6-(2-aminoethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate hydrochloride (21 mg, 98%, 0.0722 mmol) as a pale red powder. Ethyl 6-(2-aminoethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate hydrochloride (21.00 mg, 0.0722 mmol) and 3-(5-methyl-1,2,4-oxadiazol-3-yl)benzoic acid (22.12 mg, 0.1083 mmol) were dissolved in dry DMF (0.36 mL). 1-Propanephosphonic anhydride (63.82 μL, 0.1083 mmol) and triethylamine (15.25 μL, 0.1083 mmol) were added, and the reaction mixture was stirred at room temperature for 18 h. 1-Propanephosphonic anhydride (63.82 μL, 0.1083 mmol) and 3-(5-methyl-1,2,4-oxadiazol-3-yl)benzoic acid (22.12 mg, 0.1083 mmol) were added, and the mixture was stirred at room temperature for 3 hours. Purification by reverse-phase flash chromatography (eluent: 30-80% MeOH / water + 0.1% formic acid) followed by ion-exchange SCX-2 chromatography eluting with 2M NH in MeOH afforded ethyl 6-[2-[[3-(5-methyl-1,2,4-oxadiazol-3-yl)benzoyl]amino]ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate (3 mg, 9%, 0.0066 mmol) as an off-white solid. HPLC / MS m / z: 441.16 [M+H] + , Rt (AF): 2.13 min. 1H NMR (600 MHz, Methanol-d4): δ 8.52 (t, J = 1.8 Hz, 1H), 8.19 (dt, J = 7.8, 1.4 Hz, 1H), 8.01-7.96 (m, 1H), 7.61 (t, J = 7.8 Hz, 1H), 7.51 (s, 1H), 4.30 (q, J = 7.1 Hz, 2H), 3.87-3.82 (m, 2H), 3.66 (t, J = 6.6 Hz, 2H), 2.92 (t, J = 5.9 Hz, 2H), 2.86 (t, J = 6.7 Hz, 2H), 2.82-2.78 (m, 2H), 2.66 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0435] The following examples were obtained in a similar manner. Example 70: Propan-2-yl 6-[2-[[3-(5-methyl-1,2,4-oxadiazol-3-yl)benzoyl]amino]ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate [ka] Colorless powder (10mg, 37%, 0.0220mmol). HPLC / MS m / z: 455.17 [M+H] + , Rt (AF): 2.26 min. 1H NMR (600 MHz, Methanol-d4): δ 8.52 (t, J = 1.8 Hz, 1H), 8.19 (dt, J = 7.8, 1.4 Hz, 1H), 8.01-7.96 (m, 1H), 7.61 (t, J = 7.8 Hz, 1H), 7.49 (s, 1H), 5.13 (hept, J = 6.3 Hz, 1H), 3.86-3.82 (m, 2H), 3.65 (t, J = 6.6 Hz, 2H), 2.92 (t, J = 5.9 Hz, 2H), 2.86 (t, J = 6.6 Hz, 2H), 2.80 (t, J = 5.9 Hz, 2H), 2.66 (s, 3H), 1.33 (d, J = 6.3 Hz, 6H).

[0436] Example 71: tert-Butyl 6-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate [ka]

[0437] Example 71.1: 6-tert-Butoxycarbonyl-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylic acid (100.00 mg, 0.3529 mmol) was dissolved in dry THF (1.47 mL) and cooled to 0 °C. tert-Butyl-2,2,2-trichloroacetamidate (231.36 mg, 1.0588 mmol) in THF (0.2 mL) was added, followed by the dropwise addition of boron trifluoride diethyl etherate (0.11 mL, 0.8823 mmol). The reaction was allowed to warm to room temperature for 1 h. NaHCO3 and EtOAc were added, and the organic layer was washed with water and dried over MgSO4. Purification by normal phase silica column chromatography (0-35% EtOAc in cyclohexane) afforded di-tert-butyl 5,7-dihydro-4H-thieno[2,3-c]pyridine-2,6-dicarboxylate (112 mg, 93%, 0.3299 mmol) as a yellow oil. HPLC / MS m / z: 362.14 [M+Na] + , Rt (AE): 1.72 min.

[0438] Example 71.2: tert-Butyl 6-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate was prepared from di-tert-butyl 5,7-dihydro-4H-thieno[2,3-c]pyridine-2,6-dicarboxylate in a manner analogous to that shown in Example 69.

[0439] Example 71.3: tert-Butyl 6-(2-aminoethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate (32.81 mg, 0.1162 mmol), 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (22.00 mg, 0.0968 mmol) and PyBroP (55.07 mg, 0.1181 mmol) were dissolved in dry DCM (0.48 mL). DIPEA (80.11 μL, 0.4599 mmol) was added and the reaction mixture was stirred at room temperature for 2 d. Reverse phase column chromatography (eluent: 40-100% MeOH / water + 0.1% formic acid) followed by MeOH Purification by SCX-2 ion exchange chromatography eluting with ammonia in 2M afforded tert-butyl 6-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate (10 mg, 21%, 0.0203 mmol) as a colorless powder. HPLC / MS m / z: 492.21 [M+H] + , Rt (AF): 2.81 min. 1 H NMR (600 MHz, Methanol-d4): δ 8.83-8.79 (m, 1H), 8.18 (dd, J = 8.5, 1.6 Hz, 1H), 7.90 (d, J = 5.9 Hz, 1H), 7.77 (d, J = 8.5 Hz, 1H), 7.42 (s, 1H), 6.95 (dd, J = 6.0, 0.9 Hz, 1H), 3.88-3.83 (m, 2H), 3.80 (t, J = 6.4 Hz, 2H), 2.97 (t, J = 6.4 Hz, 2H), 2.93 (t, J = 5.8 Hz, 2H), 2.82-2.78 (m, 2H), 2.66 (s, 3H), 1.55 (s, 9H).

[0440] Example 72: Ethyl 5-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate [ka]

[0441] Example 72.1: 4,5,6,7-Tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylic acid hydrochloride (600.00 mg, 2.9465 mmol) was placed in a flask. Di-tert-butyl dicarbonate (662.97 mg, 2.9465 mmol) in MeCN (11.00 mL) was added, followed by triethylamine (0.41 mL, 2.9465 mmol). The reaction mixture was stirred at 40° C. for 40 min. The crude 5-tert-butoxycarbonyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylic acid (1337 mg) was used in the next step without purification. 5-tert-Butoxycarbonyl-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylic acid (700.00 mg, 1.7287 mmol), 4-dimethylaminopyridine (42.24 mg, 0.3457 mmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (497.09 mg, 2.593 mmol) were dissolved in a mixture of DMF (2.88 mL) and ethanol (2.88 mL). The reaction mixture was stirred at 50 °C for 4 h. The reaction mixture was concentrated in vacuo. EtOAc and aqueous NH4Cl solution were added, and the organic layer was washed again with aqueous NH4Cl, dried over MgSO4, and concentrated in vacuo to give O5-tert-butyl O2-ethyl 6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2,5-dicarboxylate (400 mg, 78%, 1.3544 mmol) as a colorless oil. 1H NMR (500 MHz, Chloroform-d): δ 6.63 (t, J = 0.9 Hz, 1H), 4.68 (s, 2H), 4.41 (q, J = 7.1 Hz, 2H), 4.27 (t, J = 5.5 Hz, 2H), 3.91 (t, J = 5.5 Hz, 2H), 1.51 (s, 9H), 1.40 (t, J = 7.1 Hz, 3H).

[0442] Example 72.2: O5-tert-Butyl O2-ethyl 6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2,5-dicarboxylate (400.00 mg, 1.3544 mmol) was dissolved in dry 1,4-dioxane (4.51 mL). Hydrogen chloride (4 M in 1,4-dioxane) (1.69 mL, 6.7721 mmol) was added, and the reaction mixture was stirred at room temperature for 4.5 h. Additional hydrogen chloride (4 M in 1,4-dioxane) (1.69 mL, 6.7721 mmol) was added, and the reaction mixture was stirred for 16 h. The solvent was removed in vacuo to give crude ethyl 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate hydrochloride (360 mg, 115%, 1.5539 mmol) as a colorless powder. HPLC / MS m / z: 196.11 [M+H] + , Rt (AI): 0.27 min.

[0443] Example 72.3: Ethyl 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate hydrochloride (150.00 mg, 0.6474 mmol) was dissolved in CHCl3 (3.24 mL). Triethylamine (0.09 mL, 0.6474 mmol) was added and the reaction mixture was cooled to 0 °C. 1-Nitroethene (1 M in xylene) (0.65 mL, 0.6474 mmol) was added and stirred at 0 °C for 50 min. The mixture was concentrated in vacuo. Purification by column chromatography (eluent: 0–10% MeOH in EtOAc) afforded ethyl 5-(2-nitroethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (117 mg, 67%, 0.4361 mmol) as a colorless oil that solidified on standing. 1 H NMR (500 MHz, DMSO-d6): δ 6.51 (s, 1H), 4.83-4.75 (m, 2H), 4.24 (q, J = 7.1 Hz, 2H), 4.11 (t, J = 5.6 Hz, 2H), 3.77 (s, 2H), 3.15-3.09 (m, 2H), 3.05-3.00 (m, 2H), 1.26 (t, J = 7.1 Hz, 3H).

[0444] Example 72.4: Ethyl 5-(2-nitroethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (90.00 mg, 0.3355 mmol) was dissolved in ethanol (1.68 mL). Palladium on carbon (35.70 mg) was added and the atmosphere was replaced with hydrogen. The reaction mixture was stirred at room temperature for 22 h. The reaction mixture was filtered through Celite and rinsed with EtOH. The solvent was removed under reduced pressure. Purification by reverse-phase flash chromatography (eluent: 10-60% MeOH / water + 0.1% formic acid) followed by ion-exchange SCX-2 chromatography eluting with 2M NH3 in MeOH afforded ethyl 5-(2-aminoethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (18 mg, 23%). HPLC / MS m / z: 239.15 [M+H]+ , Rt (AI): 0.61 min.

[0445] Example 72.5: Ethyl 5-(2-aminoethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (12.00 mg, 0.0504 mmol), 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (22.89 mg, 0.1007 mmol), and bromotri(pyrrolidino)phosphonium hexafluorophosphate (35.21 mg, 0.0755 mmol) were dissolved in dry DCM (0.50 mL) in a 0.5-2 mL microwave vial. DIPEA (40.35 μL, 0.2317 mmol) was added, and the reaction mixture was irradiated at 60 °C for 1 h. The solvent was removed in vacuo. Purification by reverse-phase flash chromatography (eluent: 5-60% MeCN / water + 0.1% formic acid), followed by ion-exchange SCX-2 chromatography eluting with 2M NH in MeOH, afforded ethyl 5-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (6 mg, 25%, 0.0127 mmol) as a colorless powder. HPLC / MS m / z: 448.21 [M+H] + , Rt (AD): 2.12 min. 1H NMR (600 MHz, Methanol-d4): δ 8.81-8.77 (m, 1H), 8.19 (dd, J = 8.5, 1.6 Hz, 1H), 7.91 (d, J = 5.9 Hz, 1H), 7.78 (d, J = 8.5 Hz, 1H), 6.96 (dd, J = 6.0, 0.9 Hz, 1H), 6.56 (s, 1H), 4.33 (q, J = 7.1 Hz, 2H), 4.25 (t, J = 5.6 Hz, 2H), 3.88 (s, 2H), 3.80 (t, J = 6.3 Hz, 2H), 3.18-3.12 (m, 2H), 2.98 (t, J = 6.3 Hz, 2H), 2.66 (s, 3H), 1.35 (t, J = 7.1 Hz, 3H).

[0446] Example 73: Propan-2-yl 5-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate [ka]

[0447] Example 73.1: Propan-2-yl 5-(2-aminoethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate was prepared by a procedure similar to Example 72.

[0448] Example 73.2: Propan-2-yl 5-(2-nitroethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (110.00 mg, 0.3897 mmol), zinc (127.38 mg, 1.9483 mmol), and ammonium chloride (104.21 mg, 1.9483 mmol) were suspended in a mixture of THF (1.50 mL) / water (0.50 mL) / MeOH (0.50 mL). The reaction mixture was stirred at room temperature for 2 h. Additional zinc (127.38 mg, 1.9483 mmol) and ammonium chloride (104.21 mg, 1.9483 mmol) were added and stirred for an additional 1 h. The mixture was filtered through Celite. The filtrate was passed through a 5 g NH ion exchange column and rinsed with EtOAc, then with a 1:1 mixture of DCM:MeOH. The filtrate was concentrated to give propan-2-yl 5-(2-aminoethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (60 mg, 61%, 0.2378 mmol) as a colorless oil. HPLC / MS m / z: 253.17 [M+H] + , Rt (AI): 0.74 min.

[0449] Example 73.3: In a procedure similar to Example 72.5, propan-2-yl 5-(2-aminoethyl)-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (30.00 mg, 0.1189 mmol) was used to give propan-2-yl 5-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6,7-dihydro-4H-pyrazolo[1,5-a]pyrazine-2-carboxylate (5 mg, 9%, 0.0103 mmol) as a colorless powder. HPLC / MS m / z: 462.23 [M+H] + , Rt (AJ): 2.03 min. 1H NMR (600 MHz, Methanol-d4): δ 8.81-8.78 (m, 1H), 8.20 (dd, J = 8.5, 1.6 Hz, 1H), 7.91 (d, J = 5.9 Hz, 1H), 7.79 (d, J = 8.5 Hz, 1H), 6.97 (dd, J = 5.9, 0.9 Hz, 1H), 6.57-6.53 (m, 1H), 5.19 (hept, J = 6.3 Hz, 1H), 4.26 (t, J = 5.6 Hz, 2H), 3.88 (s, 2H), 3.80 (t, J = 6.3 Hz, 2H), 3.15 (t, J = 5.6 Hz, 2H), 2.99 (t, J = 6.3 Hz, 2H), 2.66 (s, 3H), 1.34 (d, J = 6.3 Hz, 6H).

[0450] Example 74: 6-(5,5-dimethyl-4H-oxazol-2-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one [ka] 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxoisoindoline-5-carbonitrile [Example 59.2] (42.00 mg, 0.1023 mmol) and zinc chloride (2.79 mg, 0.0205 mmol) were combined in a microwave vial. The capped vial was evacuated for 10 min. The reactor was then flushed with argon, and anhydrous toluene (0.10 mL) and 1-amino-2-methylpropan-2-ol (0.01 mL, 0.1228 mmol) were added. The reaction mixture was heated at 110 °C overnight. More toluene (1 mL) and 1-amino-2-methylpropan-2-ol (10 equiv.) were added, and the reaction mixture was heated at 150 °C for 10 h by microwave irradiation. The crude material was purified by preparative HPLC (Teledyne AccqPrep, 67-74% MeOH in water, pH 10, 20 min). Further purification by column chromatography (eluent: 0-15% EtOAc in MeOH) afforded 6-(5,5-dimethyl-4H-oxazol-2-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one (14 mg, 28%, 0.0283 mmol) as a pink solid. HPLC / MS m / z: 483.214 [M+H] + , Rt (AJ): 1.92 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.82 (d, J = 1.5 Hz, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.05-8.01 (m, 2H), 7.99 (d, J = 1.4 Hz, 1H), 7.94 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.71-7.66 (m, 1H), 6.96-6.92 (m, 1H), 4.65 (s, 2H), 3.88-3.84 (m, 2H), 3.82-3.78 (m, 2H), 3.71 (s, 2H), 2.67 (s, 3H), 1.43 (s, 6H).

[0451] Example 75: 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6-(5-propyl-1,2,4-oxadiazol-3-yl)isoindolin-1-one [ka]

[0452] Example 75.1: 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxoisoindoline-5-carbonitrile [Example 59.2] (129.00 mg, 0.3143 mmol), hydroxylamine hydrochloride (24.03 mg, 0.3457 mmol), and triethylamine (0.07 mL, 0.4715 mmol) were mixed in [bmim]OAc (0.63 mL) under argon. The reaction mixture was heated at 80 °C for 1 h. The reaction mixture was cooled to ambient temperature and mixed with EtOAc (30 mL). The organic layer was mixed with saturated NaCl (50 mL) and separated again. The aqueous layer was further extracted with EtOAc (2 x 30 mL). The combined organic layers were washed with saturated NaCl (3 x 20 mL) and then concentrated under reduced pressure through a pad of anhydrous MgSO without filtration to retain the undissolved product. The crude N-hydroxy-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxoisoindoline-5-carboxamidine (88 mg, 63%, 0.1984 mmol) was used in the next step without further purification. HPLC / MS m / z: 444.175 [M+H]+, Rt (AD): 1.25 min.

[0453] Example 75.2: N-hydroxy-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3-oxo-isoindoline-5-carboxamidine (40.00 mg, 0.0902 mmol), butyric anhydride (0.03 mL, 0.1804 mmol), and anhydrous MeCN (1.00 mL) were mixed in a microwave vial under argon. The reaction mixture was heated at 160 °C by microwave irradiation for 10 min. Volatiles were removed under reduced pressure. The crude product was directly purified by preparative HPLC (AccqPrep, 30-43% MeOH in water, pH 3, 26 min). Purification by ion-exchange SCX-2 chromatography eluting with 2M NH in MeOH gave 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6-(5-propyl-1,2,4-oxadiazol-3-yl)isoindolin-1-one (28 mg, 62%, 0.0559 mmol) as a colorless powder. HPLC / MS m / z: 496.210 [M+H] + , Rt (AJ): 2.33 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.84-8.82 (m, 1H), 8.20 (dd, J = 7.9, 1.6 Hz, 1H), 8.17 (dd, J = 1.5, 0.8 Hz, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.04 (t, J = 5.6 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.79 (dd, J = 7.9, 0.9 Hz, 1H), 6.95 (dd, J = 5.9, 0.9 Hz, 1H), 4.68 (s, 2H), 3.90-3.86 (m, 2H), 3.84-3.79 (m, 2H), 2.99 (t, J = 7.4 Hz, 2H), 2.67 (s, 3H), 1.82 (h, J = 7.4 Hz, 2H), 0.99 (t, J = 7.4 Hz, 3H).

[0454] The following examples were prepared in a similar manner. Example 76: 6-(5-ethyl-1,2,4-oxadiazol-3-yl)-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one [ka] HPLC / MS m / z: 482.195 [M+H] + , Rt (AJ): 2.18 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.85-8.82 (m, 1H), 8.20 (dd, J = 7.8, 1.6 Hz, 1H), 8.17 (dd, J = 1.5, 0.8 Hz, 1H), 8.14 (dd, J = 8.5, 1.6 Hz, 1H), 8.04 (t, J = 5.6 Hz, 1H), 7.95 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.79 (dd, J = 7.8, 0.9 Hz, 1H), 6.95 (dd, J = 5.9, 0.8 Hz, 1H), 4.68 (s, 2H), 3.90-3.86 (m, 2H), 3.84-3.79 (m, 2H), 3.02 (q, J = 7.6 Hz, 2H), 2.67 (s, 3H), 1.35 (t, J = 7.6 Hz, 3H).

[0455] Example 77: Propan-2-yl 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1,3-dioxo-isoindoline-5-carboxylate [ka]

[0456] Example 77.1: A solution of 3-(1-chloro-7-isoquinolyl)-5-methyl-1,2,4-oxadiazole [Intermediate A4] (1000.00 mg, 4.0707 mmol) and ethylenediamine (5.44 mL, 81.413 mmol) in NMP (5.00 mL) was placed in a 10-20 mL microwave vial and heated at 160 °C under microwave irradiation for 1 h. Saturated bicarbonate solution (100 mL) was added and the product was extracted with EtOAc (3 x 50 mL). The combined organic layers were dried over magnesium sulfate and concentrated under reduced pressure to give N'-[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]ethane-1,2-diamine (982 mg, 90%, 3.6465 mmol) as a yellow solid. HPLC / MS m / z: 270.136 [M+H] + , Rt (AE): 0.95 min.

[0457] Example 77.2: N'-[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]ethane-1,2-diamine (500.00 mg, 1.8567 mmol) and 4-bromophthalic anhydride (442.56 mg, 1.9495 mmol) were mixed in glacial acetic acid (3.71 mL, 0.5000 M) in a microwave vial under argon. The reaction mixture was heated at 150 °C by microwave irradiation for 1 h. Purification by silica gel normal phase column chromatography (eluent: 40-100% EtOAc in cyclohexane) afforded 5-bromo-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindoline-1,3-dione (761 mg, 85%, 1.5815 mmol) as a pale yellow powder. HPLC / MS m / z: 478.050 [M+H] + , Rt (AE): 2.18 min.

[0458] Example 77.3: 5-Bromo-2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]isoindoline-1,3-dione (50.00 mg, 0.1045 mmol) and XantPhos Pd G4 (1.01 mg, 0.0010 mmol) were combined in a microwave vial. The capped vial was evacuated and fitted with a balloon filled with carbon monoxide. Anhydrous 1,4-dioxane (0.52 mL), anhydrous iPrOH (0.40 mL, 5.2268 mmol), and DIPEA (0.04 mL, 0.2091 mmol) were added, and the reaction mixture was heated at 80 °C for 6 h. The reaction mixture was concentrated under reduced pressure, dissolved in DMSO, and purified by preparative HPLC (AccqPrep, 55-65% ACN in water, pH 10, 20 min) to give propan-2-yl 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1,3-dioxo-isoindoline-5-carboxylate (16 mg, 31%, 0.0323 mmol) as a pale yellow solid. HPLC / MS m / z: 486.178 [M+H] + , Rt (AJ): 2.37 min. 1 H NMR (600 MHz, DMSO-d6): δ 8.70 (d, J = 1.6 Hz, 1H), 8.33 (dd, J = 7.7, 1.4 Hz, 1H), 8.16 (d, J = 1.4 Hz, 1H), 8.13 (dd, J = 8.5, 1.6 Hz, 1H), 7.98 (t, J = 6.0 Hz, 2H), 7.96 (d, J = 7.8 Hz, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.81 (d, J = 5.8 Hz, 2H), 6.90 (d, J = 5.7 Hz, 1H), 5.18 (hept, J = 6.2 Hz, 1H), 3.95-3.91 (m, 2H), 3.83-3.77 (m, 2H), 2.65 (s, 3H), 1.35 (d, J = 6.3 Hz, 6H).

[0459] Example 78: Ethyl 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1-oxo-3H-pyrrolo[3,4-c]pyridine-6-carboxylate [ka]

[0460] Example 78.1: 2-Chloro-5-methylisonicotinic acid ethyl ester (500.00 mg, 2.5046 mmol) was dissolved in anhydrous DCE (25.05 mL) under argon. AIBN (41.13 mg, 0.2505 mmol) and NBS (445.77 mg, 2.5046 mmol) were added, and the reaction mixture was heated to reflux for 2 h. The reaction mixture was cooled to ambient temperature. Purification by silica gel normal phase column chromatography (eluent: 0-30% EtOAc in cyclohexane) afforded ethyl 5-(bromomethyl)-2-chloro-pyridine-4-carboxylate (577 mg, 62%, 1.5537 mmol) as a colorless oil. HPLC / MS m / z: 277.958 [M+H] + , Rt (AJ): 2.57 min.

[0461] Example 78.2: Ethyl 5-(bromomethyl)-2-chloro-pyridine-4-carboxylate (570.00 mg, 1.5348 mmol), DIPEA (0.53 mL, 3.0697 mmol), and 1-Boc-ethylenediamine (0.26 mL, 1.6116 mmol) were mixed in anhydrous MeCN (18.32 mL) under argon and heated to reflux for 30 min. The reaction mixture was cooled to ambient temperature. Purification by silica gel normal-phase column chromatography (eluent: 40 to 100% EtOAc in cyclohexane) afforded tert-butyl N-[2-(6-chloro-1-oxo-3H-pyrrolo[3,4-c]pyridin-2-yl)ethyl]carbamate (449 mg, 94%, 1.4402 mmol) as a colorless amorphous solid. HPLC / MS m / z: 312.111 [M+H] +, Rt (AJ): 2.12 min.

[0462] Example 78.3: tert-Butyl N-[2-(6-chloro-1-oxo-3H-pyrrolo[3,4-c]pyridin-2-yl)ethyl]carbamate (150.00 mg, 0.4811 mmol) and Pd(dppf)Cl₂·DCM (41.32 mg, 0.0481 mmol) were combined in a microwave vial. The vial was capped, evacuated, and then fitted with a balloon filled with carbon monoxide. Anhydrous DMF (0.50 mL), absolute ethanol (0.50 mL, 8.5631 mmol), and triethylamine (0.14 mL, 0.9623 mmol) were added, and the reaction mixture was heated at 60 °C for 24 h. Purification by silica gel normal phase column chromatography (eluent: 40-100% EtOAc in cyclohexane) afforded ethyl 2-[2-(tert-butoxycarbonylamino)ethyl]-1-oxo-3H-pyrrolo[3,4-c]pyridine-6-carboxylate (73 mg, 43%, 0.2089 mmol) as an amorphous solid. HPLC / MS m / z: 350.172 [M+H] + , Rt (AJ): 2.13 min.

[0463] Example 78.4: Ethyl 2-[2-(tert-butoxycarbonylamino)ethyl]-1-oxo-3H-pyrrolo[3,4-c]pyridine-6-carboxylate (73.00 mg, 0.2089 mmol) was mixed with 4 M HCl in 1,4-dioxane (5.22 mL, 20.894 mmol) and anhydrous 1,4-dioxane (5.00 mL) at room temperature under argon and stirred for 16 h. The volatiles were removed under reduced pressure. The crude material was dissolved in MeOH and filtered through a 1 g SCX-2 column. The product was released with ammonia in MeOH / EtOH (7 M ammonia in MeOH diluted 1:4 with ethanol to reduce the possibility of transesterification) to give ethyl 2-(2-aminoethyl)-1-oxo-3H-pyrrolo[3,4-c]pyridine-6-carboxylate (48 mg, 92%, 0.1926 mmol) as a light brown oil. HPLC / MS m / z: 250.119 [M+H] + , Rt (AJ): 0.32 min.

[0464] Example 78.5: Ethyl 2-(2-aminoethyl)-1-oxo-3H-pyrrolo[3,4-c]pyridine-6-carboxylate (48.00 mg, 0.1926 mmol), 5-methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (48.13 mg, 0.2118 mmol), PyBrop (98.75 mg, 0.2118 mmol), DIPEA (0.13 mL, 0.7221 mmol), and anhydrous DCM (1.00 mL) were placed in a microwave vial under argon at room temperature. The reaction mixture was heated at 60 °C by microwave irradiation for 1 h. The volatiles were removed under reduced pressure. Silica gel normal phase column chromatography (eluent: 0–15% MeOH in EtOAc) followed by NH in MeOH / EtOH 3(Purification by ion-exchange SCX-2 chromatography eluting with 7M ammonia in MeOH diluted 1:4 with ethanol to reduce the possibility of transesterification. Further purification by reverse-phase flash chromatography (eluent: 20-60% MeOH in water) followed by ion-exchange SCX-2 chromatography eluting with ammonia solution afforded ethyl 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-1-oxo-3H-pyrrolo[3,4-c]pyridine-6-carboxylate (29 mg, 33%, 0.0632 mmol) as an off-white amorphous solid. HPLC / MS m / z: 459.178 [M+H] + , Rt (AJ): 1.89 min. 1 H NMR (600 MHz, DMSO-d6): δ 9.03 (d, J = 1.1 Hz, 1H), 8.80-8.79 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.13 (d, J = 1.1 Hz, 1H), 8.01 (t, J = 5.7 Hz, 1H), 7.91 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.94 (dd, J = 5.9, 0.8 Hz, 1H), 4.79 (s, 2H), 4.36 (q, J = 7.1 Hz, 2H), 3.91-3.88 (m, 2H), 3.85-3.80 (m, 2H), 2.67 (s, 3H), 1.34 (t, J = 7.1 Hz, 3H).

[0465] Example 79: 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6-(propylamino)isoindolin-1-one [ka]

[0466] Example 79.1: tert-Butyl N-[2-(6-bromo-1-oxo-isoindolin-2-yl)ethyl]carbamate [Example 56.1] (200.00 mg, 0.5630 mmol), cesium carbonate (369.15 mg, 1.126 mmol), BrettPhos Pd G3 (5.10 mg, 0.0056 mmol), and BrettPhos (3.02 mg, 0.0056 mmol) were mixed in a microwave vial under argon. 2-Methyl-2-butanol (2.82 mL) and propylamine (0.06 mL, 0.6756 mmol) were added, and the reaction mixture was heated at 100 °C for 1.5 h. The reaction mixture was cooled to ambient temperature. Purification by column chromatography (eluent: 40-100% EtOAc in cyclohexane) yielded tert-butyl N-[2-[1-oxo-6-(propylamino)isoindolin-2-yl]ethyl]carbamate (149 mg, 79%, 0.4469 mmol). HPLC / MS m / z: 334.212 [M+H] + , Rt (AJ): 2.30 min.

[0467] Example 79.2: A procedure similar to that used in Examples 78.4 and 78.5 gave 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6-(propylamino)isoindolin-1-one (29 mg, 0.0633 mmol) as an off-white solid. HPLC / MS m / z: 443.220 [M+H] + , Rt (AJ): 2.04 min. 1H NMR (600 MHz, DMSO-d6): δ 8.86-8.83 (m, 1H), 8.14 (dd, J = 8.5, 1.5 Hz, 1H), 8.03 (t, J = 5.5 Hz, 1H), 7.96 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.22 (d, J = 8.2 Hz, 1H), 6.95 (d, J = 5.7 Hz, 1H), 6.78 (dd, J = 8.2, 2.3 Hz, 1H), 6.72 (d, J = 2.2 Hz, 1H), 5.80 (t, J = 5.5 Hz, 1H), 4.39 (s, 2H), 3.82-3.78 (m, 2H), 3.78-3.73 (m, 2H), 2.97 (td, J = 7.1, 5.5 Hz, 2H), 2.68 (s, 3H), 1.60-1.50 (m, 2H), 0.93 (t, J = 7.4 Hz, 3H).

[0468] Example 80: Ethyl 6-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-7-oxo-5H-pyrrolo[3,4-b]pyridine-2-carboxylate [ka]

[0469] Example 80.1: Methyl 6-chloro-3-methyl-pyridine-2-carboxylate (1000.00 mg, 5.3876 mmol) was dissolved in anhydrous DCE (53.88 mL) under argon. AIBN (88.47 mg, 0.5388 mmol) and NBS (958.89 mg, 5.3876 mmol) were added, and the reaction mixture was heated to reflux for 2 h. The reaction mixture was cooled to ambient temperature. Purification by silica gel normal phase column chromatography (eluent: 0-30% EtOAc in cyclohexane) afforded methyl 3-(bromomethyl)-6-chloro-pyridine-2-carboxylate (1.12 g, 79%, 4.2457 mmol) as a colorless crystalline solid. HPLC / MS m / z: 287.986 [M+Na] + , Rt (AJ): 2.07 min.

[0470] Example 80.2: Methyl 3-(bromomethyl)-6-chloropyridine-2-carboxylate (1.12 g, 3.3875 mmol), DIPEA (1.18 mL, 6.775 mmol), and 1-Boc-ethylenediamine (0.56 mL, 3.5569 mmol) were mixed in anhydrous MeCN (33.88 mL) under argon and heated to reflux for 1.5 h. The reaction mixture was cooled to ambient temperature. Purification by silica gel normal-phase column chromatography (eluent: 0-100% EtOAc in cyclohexane) afforded tert-butyl N-[2-(2-chloro-7-oxo-5H-pyrrolo[3,4-b]pyridin-6-yl)ethyl]carbamate (748 mg, 71%, 2.3993 mmol) as a colorless crystalline solid. HPLC / MS m / z: 334.170 [M+H] + , Rt (AJ): 1.99 min.

[0471] Example 80.3: tert-Butyl N-[2-(2-chloro-7-oxo-5H-pyrrolo[3,4-b]pyridin-6-yl)ethyl]carbamate (150.00 mg, 0.4811 mmol), palladium acetate (2.18 mg, 0.0096 mmol), and XantPhos (11.14 mg, 0.0192 mmol) were combined in a microwave vial. The vial was capped, flushed with argon, evacuated, and then fitted with a balloon filled with carbon monoxide. Anhydrous 1,4-dioxane (0.96 mL), DIPEA (0.17 mL, 0.9623 mmol), and absolute ethanol (0.14 mL, 2.4057 mmol) were added, and the reaction mixture was heated at 90 °C for 72 h. Purification by silica gel normal phase column chromatography (eluent: 0-10% MeOH in EtOAc) afforded ethyl 6-[2-(tert-butoxycarbonylamino)ethyl]-7-oxo-5H-pyrrolo[3,4-b]pyridine-2-carboxylate (41 mg, 24%, 0.1174 mmol) as an amorphous solid. HPLC / MS m / z: 350.247 [M+H] + , Rt (AI): 1.22 min.

[0472] Example 80.4: Ethyl 6-[2-(tert-butoxycarbonylamino)ethyl]-7-oxo-5H-pyrrolo[3,4-b]pyridine-2-carboxylate (41.00 mg, 0.1174 mmol) was mixed with 4 M HCl in 1,4-dioxane (1.47 mL, 5.8675 mmol) and anhydrous 1,4-dioxane (3.00 mL) at room temperature under argon and stirred for 2 h. The volatiles were removed under reduced pressure. The crude material was dissolved in MeOH and filtered through a 1 g SCX-2 column. The product was released with 2 M ammonia in MeOH to give ethyl 6-(2-aminoethyl)-7-oxo-5H-pyrrolo[3,4-b]pyridine-2-carboxylate (32 mg, 88%, 0.1027 mmol) as an off-white solid. HPLC / MS m / z: 250.184 [M+H] + , Rt (AI): 0.43 min.

[0473] Example 80.5: Ethyl 6-(2-aminoethyl)-7-oxo-5H-pyrrolo[3,4-b]pyridine-2-carboxylate (30.00 mg, 0.0963 mmol), 5-methyl-3-(2-oxideisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (26.25 mg, 0.1155 mmol), PyBrop (53.86 mg, 0.1155 mmol), DIPEA (0.06 mL, 0.3611 mmol), and anhydrous DCM (0.50 mL) were placed in a microwave vial under argon at room temperature. The reaction mixture was heated at 60 °C by microwave irradiation for 1 h. The volatiles were removed under reduced pressure. Purification by ion-exchange SCX-2 chromatography eluting with reverse-phase flash (eluent: 20-60% methanol / water + 0.1% formic acid), followed by MeOH / EtOH (7 M ammonia in MeOH diluted 1:4 with ethanol to reduce the possibility of transesterification). Further purification by column chromatography (eluent: 0-20% MeOH in EtOAc) afforded ethyl 6-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-7-oxo-5H-pyrrolo[3,4-b]pyridine-2-carboxylate (9.3 mg, 20%, 0.0194 mmol) as an amorphous solid. HPLC / MS m / z: 459.174 [M+H] + , Rt (AJ): 1.84 min. 1H NMR (600 MHz, DMSO-d6): δ 8.82-8.79 (m, 1H), 8.25 (d, J = 8.0 Hz, 1H), 8.20 (d, J = 7.9 Hz, 1H), 8.14 (dd, J = 8.5, 1.6 Hz, 1H), 8.03 (t, J = 5.7 Hz, 1H), 7.92 (d, J = 5.7 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 6.94 (dd, J = 5.9, 0.9 Hz, 1H), 4.69 (s, 2H), 4....

Claims

1. Formula I, 【Chemical 1】 During the ceremony, W is 【Chemistry 2】 【Chemistry 3】 represents R 1 No 2 , COOA, A, OA, NHA, NHCOA, CONHA, CONA 2 , COA or R 4 represents R 2 teeth, 【Chemistry 4】 represents R 3 represents H or A, R 4 represents H, oxadiazolyl, tetrazolyl, pyrazolyl, oxazolyl or isoxazolyl, which is unsubstituted or substituted by unbranched or branched alkyl having 1 to 4 C atoms, A denotes unbranched or branched alkyl or cycloalkyl having 1 to 10 C atoms, where two adjacent CH groups and / or CH 2 The group may form a double bond and wherein one or two non-adjacent CH groups and / or CH 2 The group may be replaced by N, O, and / or S atoms, and wherein 1 to 7 H atoms may be replaced by F or CI; Hal denotes F, Cl, Br, or I; and physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers thereof, and mixtures thereof in any proportion.

2. W is 【Chemistry 5】 and R 1 , R 2 , R 3 , R 4 and A has the meaning as in claim 1.

10. The compound of claim 1, and its physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any proportion.

3. W is 【Chemistry 6】 and R 1 , R 2 , R 3 , R 4 and A has the meaning as in claim 1; 10. Compounds according to claim 1 or 2, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any proportion.

4. R 1 COOA, OA or R 4 and W.R. 2 , R 3 , R 4 and A has the meaning as in claim 1; Compounds according to any one of claims 1 to 3, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any proportion.

5. R 1 represents COOA, and W.R. 2 , R 3 , R 4 and A has the meaning as in claim 1; Compounds according to any one of claims 1 to 4, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any proportion.

6. R 2 teeth, 【Chemistry 7】 and W.R. 1 , R 3 , R 4 and A has the meaning as in claim 1; Compounds according to any one of claims 1 to 5, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any proportion.

7. R 2 teeth, 【Chemistry 8】 and W.R. 1 , R 3 , R 4 and A has the meaning as in claim 1; Compounds according to any one of claims 1 to 6, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio.

8. R 4 represents oxadiazolyl, oxazolyl, or tetrazolyl, and W.R. 1 , R 2 , R 3 and A has the meaning as in claim 1. Compounds according to any one of claims 1 to 7, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio.

9. R 4 represents oxadiazolyl, and W.R. 1 , R 2 , R 3 and A has the meaning as in claim 1. Compounds according to any one of claims 1 to 8, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio.

10. A compound selected from the group consisting of: 【Table 1-1】 【Table 1-2】 【Table 1-3】 【Table 1-4】 【Table 1-5】 【Table 1-6】 【Table 1-7】 【Table 1-8】 【Table 1-9】 and their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any proportion.

11. Pharmaceutical preparations comprising at least one compound according to any one of claims 1 to 10, and / or their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio.

12. 12. The pharmaceutical preparation of claim 11, further comprising an excipient and / or an adjuvant.

13. 13. A pharmaceutical preparation comprising at least one compound according to any one of claims 1 to 12, and / or their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, and at least one further pharmaceutically active compound.

14. A process for the preparation of pharmaceutical preparations, characterized in that the compound according to any one of claims 1 to 10, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, are brought into a suitable dosage form together with solid, liquid or semi-liquid excipients or adjuvants.

15. A medicament comprising at least one compound according to any one of claims 1 to 10, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions.

16. A medicament comprising at least one compound according to any one of claims 1 to 10, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions selected from the group consisting of hyperproliferative diseases and disorders.

17. 17. The medicament for use according to claim 16, wherein the hyperproliferative disease or disorder is cancer.

18. Cancers include acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenocortical carcinoma, bladder cancer, brain cancer, breast cancer, cervical hyperplasia, cervical cancer, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon cancer, endometrial cancer, esophageal cancer, essential thrombocytosis, genitourinary cancer, glioma, glioblastoma, hairy cell leukemia, head and neck cancer, Hodgkin's disease, Kaposi's sarcoma, lung cancer, lymphoma, malignant carcinoid cancer, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, thyroid cancer, 18. The medicament for use according to claim 17, wherein the cancer is selected from the group consisting of medullary adenoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, polycythemia vera, primary brain tumor, primary macroglobulinemia, prostate cancer, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small cell lung cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, and Wilms' tumor.

19. The following separate packs: a) an effective amount of a compound according to any one of claims 1 to 10, and / or their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, and b) an effective amount of a further pharmaceutically active compound; A set (kit) consisting of:

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