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WO2024042007A1 - Bicycles substitués utilisés comme inhibiteurs de hset - Google Patents

Bicycles substitués utilisés comme inhibiteurs de hset Download PDF

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Publication number
WO2024042007A1
WO2024042007A1 PCT/EP2023/072876 EP2023072876W WO2024042007A1 WO 2024042007 A1 WO2024042007 A1 WO 2024042007A1 EP 2023072876 W EP2023072876 W EP 2023072876W WO 2024042007 A1 WO2024042007 A1 WO 2024042007A1
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Prior art keywords
mmol
methyl
ethyl
hplc
mixture
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PCT/EP2023/072876
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English (en)
Inventor
Hans-Peter Buchstaller
Daniel Kuhn
Ian Collins
Thomas Matthews
Jan LANZ
François SAINT-DIZIER
Hugues PREVET
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Merck Patent GmbH
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Merck Patent GmbH
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Application filed by Merck Patent GmbH filed Critical Merck Patent GmbH
Priority to CN202380074493.0A priority Critical patent/CN120091998A/zh
Priority to CA3265564A priority patent/CA3265564A1/fr
Priority to JP2025511776A priority patent/JP2025529897A/ja
Priority to EP23757923.0A priority patent/EP4577537A1/fr
Priority to AU2023328866A priority patent/AU2023328866A1/en
Publication of WO2024042007A1 publication Critical patent/WO2024042007A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/04—Ortho-condensed systems

Definitions

  • the invention relates to substituted bicycles of the general formula I,
  • DNA replication followed by equal chromosome segregation, ensures the accurate transmission of the genetic information to daughter cells (Hall et al., 2003; Nigg, 2002; Zyss and Gergely, 2009).
  • centrosomes act as the dominant sites for spindle pole formation (Meunier and Vernos, 2012). Centrosome duplication is also tightly controlled and occurs simultaneously with DNA replication, thereby ensuring the generation of two functional centrosomes that form the poles of the mitotic spindle (Sharp et al., 2000).
  • MT microtubule
  • HSET encoded by KIFC1 in humans and Kifc5a in mice
  • a minus-end MT motor is of interest in cancer due to its impact on cell division (Cai et al., 2010; Goshima et al., 2005).
  • centrosomes and in particular HSET, for bipolar spindle formation has attracted much attention, although the precise role of HSET in this process remains a topic for debate (Mahoney et al., 2006; Tillement et al., 2009). Recent reports have linked centrosome amplification and high HSET expression to chromosome missegregation and aneuploidy, which are hallmarks of human cancer (Marx et al., 2009).
  • Centrosome amplification disrupts asymmetric cell division in neuroblastoma cells and causes tumorigenesis in a fly model (Basto et al., 2008), and supernumerary centrosomes are also found in most solid tumor types, forming markers for aggressiveness in breast, brain, prostate, cervix, kidney, and bladder cancers (Chan, 2011). Hence, it is increasingly apparent that supernumerary centrosomes are not only indicative of malignancy but may also drive malignant transformation (Ogden et al., 2013).
  • centrosome clustering prevents multipolar mitosis and cell death, it prolongs mitosis and increases the frequency of chromosome missegregation as a result of merotelic kinetochore attachments (Ganem et al., 2009; Kwon et al., 2008; Yang et al., 2008). Based on previous studies, centrosome clustering may prove to be the Achilles heel of cancer cells with supernumerary centrosomes (Basto et al., 2008), and a growing body of evidence suggests that inhibition of centrosome clustering could provide a new therapeutic strategy for tumors with a high incidence of centrosome amplification (Jordan and Wilson, 2004; Ogden et al., 2012).
  • HSET A key protein that is known to be crucial for centrosome clustering is HSET (Ned in flies). HSET is required by tumour cells to cluster supernumerary centrosomes (Basto et al., 2008; Kwon et al., 2008). HSET is a member of the Kinesin 14 family of MT motor proteins, which are force-generating enzymes that facilitate movement along MTs within the cell (Mountain et al., 1999) and which transport organelles, protein complexes and mRNAs along microtubules in an ATP-dependent fashion.
  • HSET is a minus-end directed motor kinesin, that cross-links and slides microtubules exerting inward forces (Walczak et al., 1997; Cai et al., 2009; Rath et al., 2012). Although the precise role of HSET in cell division is not clear, previous evidence suggests that it is essential for the survival of cancer, but not normal, cells (Ganem et al., 2009; Kwon et al., 2008).
  • High HSET expression levels are strongly correlated with metastasis of non-small cell lung cancer to the brain, pointing to an association 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 does not have a high incidence of centrosome amplification) does not inhibit bipolar spindle formation, and cells undergo normal division (Kleylein-Sohn et al., 2012; Kwon et al., 2008).
  • HSET depletion increases cell death and the frequency of multipolarity in cells with supernumerary centrosomes, but not in cells with a normal number of centrosomes.
  • HSET depletion induces spindle multi-polarity and selectively sensitizes centrosome amplified ER- breast cancer cell lines, including triple negative breast cancer (TNBC), to cell death (Patel et al., 2018).
  • TNBC triple negative breast cancer
  • Depletion of HSET was identified as inducing selective cytotoxicity in centrosome amplified cancer cells (Drosopoulos et al., 2014).
  • HSET overexpression has been correlated with poor prognosis and resistance to docetaxel in breast cancer (De et al., 2009; Li et al., 2015), is observed in ovarian adenocarcinoma patients (Pawar et al., 2014) and in numerous other cancer types (Pannu et al., 2015).
  • NSCLC non-small cell lung carcinoma
  • tumours including centrosome amplified tumours
  • cytotoxic microtubule-targeted drugs e.g. taxol, eribulin
  • these drugs typically show severe side effects and the emergence of drug resistance leading to early relapse.
  • agents targeting kinesin motor proteins e.g. Eg5 inhibitors
  • mono-polar spindles the opposite phenotype to HSET inhibition
  • target all rapidly dividing cells including bone marrow cells. Consequently, they share dose-limiting toxicities with other antimitotic therapies.
  • HSET inhibitor is anticipated to show reduced toxicity by selectively killing cells with centrosome amplification whereas cells with the normal number of centrosomes will remain unaffected (Ganem et al., 2009; Patel et al., 2015). These data together provide support for developing agents that selectively inhibit HSET to target centrosome-amplified tumours (Myers and Collins, 2016). Examples of small molecule HSET inhibitors have been described in the literature. AZ82 is an ADP/ATP competitive inhibitor shown to be selective against a panel of nine other kinesins including Eg5 (Wu et al., 2013).
  • the compounds according to the invention are highly selective and effective inhibitors of HSET and thus the compounds of the present invention can be used for the treament of hyperproliferative diseases and disorders such as cancer.
  • the invention relates to the compounds of the general formula I,
  • R 1 denotes NO 2 , COCA, A, OA, NHA, NHCOA, CONHA, CONA 2 , COA or R 4 ,
  • R 2 denotes
  • R 3 denotes H or A
  • R 4 denotes H, unsubsubstitued or with unbranched or branched alkyl with 1-4 C- atoms substituted oxadiazolyl, tetrazolyl, pyrazolyl, oxazolyl or isoxazoyl,
  • A denotes unbranched or branched alkyl or cycloalkyl with 1-10 C-atoms, wherein two adjacent CH- and/or CH2-groups may form a double bond and wherein one or two non-adjacent CH- and/or CH2-groups may be replaced by N-, O- and/or S-atoms and wherein 1-7 H-atoms may be replaced by F or Cl, Hal denotes F, Cl, Br or I and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein
  • R 1 , R 2 , R 3 , R 4 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein
  • R 1 , R 2 , R 3 , R 4 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein
  • R 1 denotes COOA, OA or R 4
  • W, R 2 , R 3 , R 4 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein
  • R 1 denotes COOA and W
  • R 2 , R 3 , R 4 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein
  • R 2 denotes and W, R 1 , R 3 , R 4 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein
  • R 2 denotes and W
  • R 1 , R 3 , R 4 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein R 4 denotes oxadiazolyl, oxazolyl or tetrazolyl and W, R 1 , R 2 , R 3 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • a preferred embodiment of the present invention are compounds according to formula I, wherein R 4 denotes oxadiazolyl and W, R 1 , R 2 , R 3 and A have the meanings as disclosed above, and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • the invention preferably relates to a compound selected from the group consisting of:
  • the invention further relates to a pharmaceutical preparation comprising one or more compounds according to the present invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios.
  • the invention also relates to a pharmaceutical preparation according to the invention of this type, comprising further excipients and/or adjuvants.
  • the invention relates to an above pharmaceutical preparation according to the invention, comprising at least one further medicament active compound.
  • compositions are taken to mean, for example, salts of the compounds of the present invention, and also so-called pro- drug compounds.
  • Prodrug compounds are taken to mean derivatives of the compounds of the present invention which have been modified by means of, for example, alkyl or acyl groups (see also amino- and hydroxyl-protecting groups below), sugars or oligopeptides and which are rapidly cleaved or liberated in the organism to form the effective molecules. These also include biodegradable polymer derivatives of the compound of the present invention, as described, for example, in Int. J. Pharm. 115 (1995), 61-67.
  • the compound of the present invention can be used in its final non-salt form.
  • the present invention also encompasses the use of the compound of the present invention in the form of its pharmaceutically acceptable salts, which can be derived from various organic and inorganic bases by procedures known in the art.
  • Pharmaceutically acceptable salt forms of the compound of the present invention are for the most part prepared by conventional methods. If the compound of the present invention contains a carboxyl group, one of its suitable salts can be formed by reacting the compound of the present invention ith a suitable base to give the corresponding base-addition salt.
  • Such bases are, 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, for example potassium ethoxide and sodium propoxide; and various organic bases, such as piperidine, diethanolamine and N-methylglutamine.
  • 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 for example potassium ethoxide and sodium propoxide
  • organic bases such as piperidine, diethanolamine and N-methylglutamine.
  • the aluminium salts of the compound of the present invetion are likewise included.
  • the base salts of the compounds of the present invention include aluminium, ammonium, calcium, copper, iron(lll), iron(ll), lithium, magnesium, man- ganese(lll), manganese(ll), potassium, sodium and zinc salts, but this is not intended to represent a restriction.
  • Salts of the compounds of the present invention which are derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, substituted amines, also including naturally occurring substituted amines, cyclic amines, and basic ion exchanger resins, for example arginine, betaine, caffeine, chloroprocaine, choline, N,N'-dibenzylethylenediamine (benzathine), dicyclohexylamine, diethanolamine, diethylamine, 2-diethylamino- ethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine
  • the pharmaceutically acceptable base-addition salts of the compound of the present invention are formed with metals or amines, such as alkali metals and alkaline-earth metals or organic amines.
  • metals are sodium, potassium, magnesium and calcium.
  • Preferred organic amines are N,N’-dibenzylethylene- diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methyl-D- glucamine and procaine.
  • the base-addition salts of the compounds of the present 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 can 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 a certain respect from the corresponding salt forms thereof with respect to certain physical properties, such as solubility in polar solvents; for the purposes of the invention, however, the salts otherwise correspond to the respective free acid forms thereof.
  • the term “pharmaceutically acceptable salt” in the present connection is taken to mean an active compound which comprises the compound of the present invention in the form of one of its salts, in particular if this salt form imparts improved pharmacokinetic properties on the active compound compared with the free form of the active compound or any other salt form of the active compound used earlier.
  • the pharmaceutically acceptable salt form of the active compound can also provide this active compound for the first time with a desired pharmacokinetic property which it did not have earlier and can even have a positive influence on the pharmacodynamics of this active compound with respect to its therapeutic efficacy in the body.
  • Solvates of the compound of the present invention are taken to mean adductions of inert solvent molecules of the compound of the present invention which form owing to their mutual attractive force.
  • Solvates are, for example, hydrates, such as monohydrates or dihydrates, or alcoholates, i.e. addition compounds with alcohols, such as, for example, with methanol or ethanol.
  • Compounds of the present invention may contain one or more centres of chirality, so that all stereoisomers, enantiomers, diastereomers, etc., of the compounds of the present inventionare also claimed in the present invention.
  • the invention also relates to the optically active forms (stereoisomers), the enantiomers, the racemates, the diastereomers and hydrates and solvates of these compounds.
  • Compounds of the present invention according to the invention may be chiral owing to their molecular structure and may accordingly occur 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 according to the invention may differ, it may be desirable to use the enantiomers. In these cases, the end product, but also even the intermediates, may be separated into enantiomeric compounds by chemical or physical measures known to the person skilled in the art or already employed as such in the synthesis.
  • compositions are taken to mean, for example, salts of the compounds according to the invention and also so-called prodrug compounds.
  • Prodrug compounds are taken to mean compounds of the present invention which have been modified with, for example, alkyl or acyl groups (see also amino- and hydroxyl-protecting groups below), sugars or oligopeptides and which are rapidly cleaved or liberated in the organism to form the 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.
  • Suitable acid-addition salts are inorganic or organic salts of all physiologically or pharmacologically acceptable acids, for example halides, in particular hydrochlorides or hydrobromides, lactates, sulfates, citrates, tartrates, maleates, fumarates, oxalates, acetates, phosphates, methylsulfonates or p-toluenesulfonates.
  • Solvates of the compounds of the present invention are taken to mean adductions of inert solvent molecules onto the compounds of the present invention which form owing to their mutual attractive force.
  • Solvates are, for example, hydrates, such as monohydrates or dihydrates, or alcoholates, i.e. addition compounds with alcohols, such as, for example, with methanol or ethanol.
  • a compound of the present invention includes isotopelabelled forms thereof.
  • An isotope-labelled form of a compound of the present invention is identical to this compound apart from 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 which differs from the atomic mass or mass number of the atom which usually occurs naturally.
  • isotopes which are readily commercially available, and which can be incorporated into a compound 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 0, 31 P, 32 P, 35 S, 18 F and 36 CI, respectively.
  • a compound of the present invention, a prodrug thereof or a pharmaceutically acceptable salt of either which contains one or more of the above-mentioned isotopes and/or other isotopes of other atoms is intended to be part of the present invention.
  • An isotope-labelled compound of the present invention can be used in a number of beneficial ways.
  • an isotope-labelled compound of the present invention into which, for example, a radioisotope, such as 3 H or 14 C, has been incorporated is suitable for medicament and/or substrate tissue distribution assays.
  • radioisotopes i.e. tritium ( 3 H) and carbon-14 ( 14 C)
  • 3 H tritium
  • 14 C carbon-14
  • Incorporation of heavier isotopes, for example deuterium ( 2 H) into a compound of the present invention has therapeutic advantages owing to the higher metabolic stability of this isotope-labelled compound. Higher metabolic stability translates directly into an increased in-vivo half-life or lower dosages, which under most circumstances would represent a preferred embodiment of the present invention.
  • An isotope-labelled compound of the present invention can usually be prepared by carrying out the procedures disclosed in the synthesis schemes and the related description, in the example part and in the preparation part in the present text, replacing a non-isotope-labelled reactant with a readily available isotope-labelled reactant.
  • deuterium ( 2 H) can also be incorporated into a compound of the present invention.
  • the primary kinetic isotope effect is a change in the rate of a chemical reaction that results from exchange of isotopic nuclei, which in turn is caused by the change in ground state energies necessary for covalent bond formation after this isotopic exchange.
  • Exchange of a heavier isotope usually results in a lowering of the ground state energy for a chemical bond and thus causes a reduction in the rate in rate-limiting bond breakage. If the bond breakage occurs in or in the vicinity of a saddle-point region along the coordinate of a multi-product reaction, the product distribution ratios can be altered substantially.
  • a compound of the present invention which has multiple potential sites of attack for oxidative metabolism, for example benzylic hydrogen atoms and hydrogen atoms bonded to a nitrogen atom, is prepared as a series of analogues in which various combinations of hydrogen atoms are replaced by deuterium atoms, so that some, most or all of these hydrogen atoms have been replaced by deuterium atoms.
  • Half-life determinations enable favourable and accurate determination of the extent to which the improvement in resistance to oxidative metabolism has improved. In this way, it is determined that the half-life of the parent compound can be extended by up to 100% as the result of deuterium-hydrogen exchange of this type.
  • the replacement of hydrogen by deuterium in a compound of the present invention can also be used to achieve a favourable modification of the metabolite spectrum of the starting compound in order to diminish or eliminate undesired toxic metabolites.
  • a toxic metabolite arises through oxidative carbonhydrogen (C-H) bond cleavage
  • C-H oxidative carbonhydrogen
  • the invention also relates to mixtures of the compounds of the present invention according to the invention, for example mixtures of two diastereomers, for example in the ratio 1:1, 1:2, 1 :3, 1 :4, 1:5, 1:10, 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.
  • the invention relates to a process for the preparation of the compounds of the present invention, characterized in that a) the base of a compound of the present invention is converted into one of its salts by treatment with an acid, or b) an acid of a compound of the present invention is converted into one of its salts by treatment with a base.
  • the starting materials or starting compounds are generally known. If they are novel, they can be prepared by methods known per se.
  • the starting materials can also be formed in situ by not isolating them from the reaction mixture, but instead immediately converting them further into the compounds of the present invention.
  • the compounds of the present invention are preferably obtained by liberating them from their functional derivatives by solvolysis, in particular by hydrolysis, or by hydrogenolysis.
  • Preferred starting materials for the solvolysis or hydrogenolysis are those which contain correspondingly protected amino, carboxyl and/or hydroxyl groups instead of one or more free amino, carboxyl and/or hydroxyl groups, preferably those which carry an amino-protecting group instead of an H atom which is connected to an N atom.
  • Preference is furthermore given to starting materials which carry a hydroxyl-protecting group instead of the H atom of a hydroxyl group.
  • Preference is also given to starting materials which carry a protected carboxyl group instead of a free carboxyl group.
  • amino-protecting group is generally known and relates to groups which are suitable for protecting (blocking) an amino group against chemical reactions, but which can easily be removed after the desired chemical reaction has been carried out elsewhere in the molecule.
  • Typical of such groups are, in particular, unsubstituted or substituted acyl groups, furthermore unsubstituted or substituted aryl (for example 2,4-dinitophenyl) or aralkyl groups (for example benzyl, 4- nitrobenzyl, triphenylmethyl). Since the amino-protecting groups are removed after the desired reaction or reaction sequence, their type and size are, in addition, not crucial, but preference is given to those having 1-20, in particular 1-8, C atoms.
  • acyl group is to be understood in the broadest sense in connection with the present process. It encompasses acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic acids or sulfonic acids and, in particular, alkoxycarbonyl, aryloxycarbonyl and especially aralkoxycarbonyl groups.
  • acyl groups are alkanoyl, such as acteyl, propionyl, buturyl, aralkanoyl, such as phenylacetyl, aroyl, such as benzoyl or toluyl, aryoxyaklkanoyl, such as phenoxyacetyl, alkyoxycarbonyyl, such as methoxycarbonyl, ethoxycarbonyl, 2,2,2- trichloroethoxycarbonyl, BOC, 2-iodoethoxycaronyl, aralkoxycarbonyl, such as CBZ, 4-methoxybenzyloxycarbonyl or FMOC.
  • Preferred acyl groups are CBZ, FMOC, benzyl and acetyl.
  • acid-protecting group or “carboxyl-protecting group” is likewise generally known and relates to groups which are suitable for protecting a -COOH group against chemical reactions, but which can easily be removed after the desired chemical reaction has been carried out elsewhere in the molecule.
  • esters instead of the free acids, for example of substituted and unsubstituted alkyl esters (such as methyl, ethyl, tert-butyl and substituted derivatives thereof), of substituted and unsubstituted benzyl esters or silyl esters, is typical.
  • the type and size of the acid-protecting groups is not crucial, but preference is given to those having 1-20, in particular 1-10, C atoms.
  • hydroxyl-protecting group is likewise generally known and relates to groups which are suitable for protecting a hydroxyl group against chemical reactions, but which can easily be 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, furthermore also alkyl groups. Their type and size of the hydroxyl-protecting groups is not crucial, but preference is given to those having 1-20, in particular 1-10, C atoms.
  • hyrdoxyl-protecting groups are, inter alia, benzyl, p-nitrobenzoyl, p-toluenesulfonyl and acetyl, where benzyl and acetyl are preferred.
  • the functional derivatives of the compounds of the present invention to be used as starting materials can be prepared by known methods of amino-acid and peptide synthesis, as described, for example, in the said standard works and patent applications.
  • the compounds of the present invention are liberated from their functional derivatives, depending on the protecting group used, for example, with the aid of strong acids, advantageously using trifluoroacetic acid or perchloric acid, but also using 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.
  • strong acids advantageously using trifluoroacetic acid or perchloric acid
  • other strong inorganic acids such as hydrochloric acid or sulfuric acid
  • strong organic acids such as trichloroacetic acid
  • sulfonic acids such as benzoyl- or p- toluenesulfonic acid.
  • the starting materials can optionally be reacted in the presence of an inert solvent.
  • 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 for 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-methylpyrrol
  • the amount of solvent is not crucial; 10 g to 500 g of solvent can preferably be added per g of the compound of the present invention to be reacted.
  • an acid-binding agent for example an alkali metal or alkaline-earth metal hydroxide, carbonate or bicarbonate or other alkali or alkaline- earth metal salts of weak acids, preferably a potassium, sodium or calcium salt, or to add an organic base, such as, for example, triethylamine, dimethylamine, pyridine or quinoline, or an excess of the amine component.
  • an acid-binding agent for example an alkali metal or alkaline-earth metal hydroxide, carbonate or bicarbonate or other alkali or alkaline- earth metal salts of weak acids, preferably a potassium, sodium or calcium salt
  • organic base such as, for example, triethylamine, dimethylamine, pyridine or quinoline, or an excess of the amine component.
  • the resultant compounds according to the invention can be separated from the corresponding solution in which they are prepared (for example by centrifugation and washing) and can be stored in another composition after separation, or they can remain directly in the preparation solution.
  • the resultant compounds according to the invention can also be taken up in desired solvents for the particular use.
  • the reaction duration depends on the reaction conditions selected. In general, the reaction duration is 0.5 hour to 10 days, preferably 1 to 24 hours. On use of a microwave, the reaction time can be reduced to values of 1 to 60 minutes.
  • the compounds of the present invention and also the starting materials for their preparation are, in addition, prepared by known methods, as described in the literature (for example in standard works, such as Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], Georg-Thieme-Verlag, Stuttgart), for example under reaction conditions which are known and suitable for the said reactions. Use can also be made here of variants known per se, which are not described here in greater detail.
  • An acid of the present invention can be converted into the associated addition salt using a base, for example by reaction of equivalent amounts of the acid and base in an inert solvent, such as ethanol, and inclusive evaporation.
  • Suitable bases for this reaction are, in particular, those which give physiologically acceptable salts.
  • the acid of the present invention can be converted into the corresponding metal salt, in particular alkali or alkaline-earth metal salt, using a base (for example sodium hydroxide, potassium hydroxide, sodium carbonate or potassium carbonate) or into the corresponding ammonium salt.
  • a base for example sodium hydroxide, potassium hydroxide, sodium carbonate or potassium carbonate
  • Organic bases which give physiologically acceptable salts, such as, for example, ethanolamine, are also suitable for this reaction.
  • a base of the present invention can be converted into the associated acid-addition salt using an acid, for example by reaction of equivalent amounts of the base and acid in an inert solvent, such as ethanol, with subsequent evaporation.
  • Suitable acids for this reaction are, in particular, those which give physiologically acceptable salts.
  • inorganic acids for example sulfuric acid, nitric acid, hydrohalic acids, such as hydrochloric acid or hydrobromic acid, phosphoric acids, such as orthophosphoric acid, sulfamic acid, furthermore organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic, mono- or polybasic carboxylic, sulfonic or sulfuric acids, for example 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-tol
  • the invention therefore furthermore relates to the use of 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.
  • the invention thus also relates, in particular, 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, including mixtures thereof in all ratios, for use in the treatment and/or prophylaxis of physiological and/or pathophysiological states.
  • physiological and/or pathophysiological states which are connected to HSET.
  • Physiological and/or pathophysiological states are taken to mean physiological and/or pathophysiological states which are medically relevant, such as, for example, diseases or illnesses and medical disorders, complaints, symptoms or complications and the like, in particular diseases.
  • the invention furthermore 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, including mixtures thereof in all ratios, for use in the treatment and/or prophylaxis of physiological and/or pathophysiological states selected from the group consisting of hyperproliferative diseases and disorders.
  • the 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, including mixtures thereof in all ratios, for use in the treatment and/or prophylaxis of physiological and/or pathophysiological states selected from the group consisting of hyperproliferative and infectious diseases and disorders, wherein the hyperproliferative disease or disorder is cancer.
  • the invention thus particularly 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, including mixtures thereof in all ratios, wherein the cancer is selected from the group consisting of acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortex cancer, bladder cancer, brain cancer, breast cancer, cervical hyperplasia, cervical cancer, chorio cancer, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon cancer, endometrial ccancer, esophageal cancer, essential thrombocytosis, genitourinary carcinoma, glioma, glioblastoma, hairy cell leukemia, head and neck carcinoma, Hodgkin's disease, Kaposi's sarcoma, lung carcinoma, lymphoma, malignant carcinoid carcinoma, malignant hypercalcemia, malignant melanoma, mal
  • the 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, including mixtures thereof in all ratios, for use in the treatment and/or prophylaxis of physiological and/or pathophysiological states 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, cirrhosis, chronic inflammatory-related disorders, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retinopathy of prematurity, granulomatosis, immune hyperproliferation associated with organ or tissue transplantation and an immunoproliferative disease or disorder selected from the group comnsisting of inflammatory bowel disease, psoriasis, rheumatoid arthritis, systemic lupus erythematosus
  • the medicaments disclosed above include a corresponding use of the compounds according to the invention for the preparation of a medicament for the treatment and/or prophylaxis of the above physiological and/or pathophysiological states. It is additionally intended that the medicaments disclosed above include a corresponding method for the treatment and/or prophylaxis of the above physiological and/or pathophysiological states in which at least one compound according to the invention is administered to a patient in need of such a treatment.
  • the compounds according to the invention preferably exhibit an advantageous biological activity which can easily be demonstrated in enzyme assays and animal experiments, as described in the examples.
  • the compounds according to the invention preferably exhibit and cause an inhibiting effect, which is usually documented by IC50 values in a suitable range, preferably in the micromolar range and more preferably in the nanomolar range.
  • the compounds according to the invention can be administered to humans or animals, in particular mammals, such as apes, dogs, cats, rats or mice, and can be used in the therapeutic treatment of the human or animal body and in the combating of the above-mentioned diseases. They can furthermore be used as diagnostic agents or as reagents.
  • compounds according to the invention can be used for the isolation and investigation of the activity or expression of HSET.
  • they are particularly suitable for use in diagnostic methods for diseases in connection with disturbed HSET activity.
  • the invention therefore furthermore relates to the use of the compounds according to the invention for the isolation and investigation of the activity or expression of HSET or as binders and inhibitors of HSET.
  • the compounds according to the invention can, for example, be radioactively labelled.
  • radioactive labels are 3 H, 14 C, 231 l and 125 l.
  • a preferred labelling method is the iodogen method (Fraker et al., 1978).
  • the compounds according to the invention can be labelled by enzymes, fluorophores and chemophores.
  • enzymes are alkaline phosphatase, - galactosidase and glucose oxidase
  • an example of a fluorophore is fluorescein
  • an example of a chemophore is luminol
  • automated detection systems for example for fluorescent colorations, are described, for example, in US 4,125,828 and US 4,207,554.
  • 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 where the partial or total inactivation of HSET could be beneficial.
  • the compounds of the present invention can be used for the preparation of pharmaceutical preparations, in particular by non-chemical methods. In this case, they are brought into a suitable dosage form together with at least one solid, liquid and/or semi-liquid excipient or adjuvant and optionally in combination with one or more further active compound(s).
  • the invention therefore furthermore relates to pharmaceutical preparations comprising at least one compound of the present invention and/or physiologically acceptable salts, derivatives, solvates and stereoisomers thereof, including mixtures thereof in all ratios.
  • the invention also relates to pharmaceutical preparations which comprise further excipients and/or adjuvants, and also to pharmaceutical preparations which comprise at least one further medicament active compound.
  • the invention also relates to a process for the preparation of a pharmaceutical preparation, characterised in that a compound of the present inventionand/or one of its physiologically acceptable salts, derivatives, solvates and stereoisomers, including mixtures thereof in all ratios, is brought into a suitable dosage form together with a solid, liquid or semi-liquid excipient or adjuvant and optionally with a further medicament active compound.
  • the pharmaceutical preparations according to the invention can be used as medicaments in human or veterinary medicine.
  • the patient or host can belong to any mammal species, for example a primate species, particularly humans; rodents, including mice, rats and hamsters; rabbits; horses, cattle, dogs, cats, etc. Animal models are of interest for experimental investigations, where they provide a model for the treatment of a human disease.
  • Suitable carrier substances are organic or inorganic substances which are suitable for enteral (for example oral), parenteral or topical administration and do not react with the novel compounds, for example water, vegetable oils (such as sunflower oil or cod-liver oil), benzyl alcohols, polyethylene glycols, gelatine, carbohydrates, such as lactose or starch, magnesium stearate, talc, lanolin or vaseline. Owing to his expert knowledge, the person skilled in the art is familiar which adjuvants are suitable for the desired medicament formulation.
  • solvents for example water, physiological saline solution or alcohols, such as, for example, ethanol, propanol or glycerol, sugar solutions, such as glucose or mannitol solutions, or a mixture of the said solvents, gel formers, tablet assistants and other activeingredient carriers
  • lubricants for example water, physiological saline solution or alcohols, such as, for example, ethanol, propanol or glycerol
  • sugar solutions such as glucose or mannitol solutions
  • gel formers such as mannitol solutions
  • a mixture of the said solvents gel formers, tablet assistants and other activeingredient carriers
  • lubricants for example water, physiological saline solution or alcohols, such as, for example, ethanol, propanol or glycerol
  • sugar solutions such as glucose or mannitol solutions
  • emulsifiers for example, emulsifiers, salts for influencing the osmotic pressure
  • antioxidants for dispers
  • preparations or medicaments according to the invention may comprise one or more further active compounds and/or one or more action enhancers (adjuvants).
  • “pharmaceutically tolerated” relates to medicaments, precipitation reagents, excipients, adjuvants, stabilisers, solvents and other agents which facilitate the administration of the pharmaceutical preparations obtained therefrom to a mammal without undesired physiological side effects, such as, for example, nausea, dizziness, digestion problems or the like.
  • the compounds according to the invention preferably have the advantage that direct use is possible and further purification steps for the removal of toxicologically unacceptable agents, such as, for example, high concentrations of organic solvents or other toxicologically unacceptable adjuvants, are thus unnecessary before use of the compounds according to the invention in pharmaceutical formulations.
  • the invention particularly preferably also relates to pharmaceutical preparations comprising at least one compound according to the invention in precipitated noncrystalline, precipitated crystalline or in dissolved or suspended form, and optionally excipients and/or adjuvants and/or further pharmaceutical active compounds.
  • the compounds according to the invention preferably enable the preparation of highly concentrated formulations without unfavourable, undesired aggregation of the compounds according to the invention occurring.
  • ready-to-use solutions having a high active-ingredient content can be prepared with the aid of compounds according to the invention with aqueous solvents or in aqueous media.
  • the compounds and/or physiologically acceptable salts and solvates thereof can also be lyophilised and the resultant lyophilisates used, for example, for the preparation of injection preparations.
  • Aqueous preparations can be prepared by dissolving or suspending compounds according to the invention in an aqueous solution and optionally adding adjuvants.
  • defined volumes of stock solutions comprising the said further adjuvants in defined concentration are advantageously added to a solution or suspension having a defined concentration of compounds according to the invention, and the mixture is optionally diluted with water to the pre-calculated concentration.
  • the adjuvants can be added in solid form. The amounts of stock solutions and/or water which are necessary in each case can subsequently be added to the aqueous solution or suspension obtained.
  • Compounds according to the invention can also advantageously be dissolved or suspended directly in a solution comprising all further adjuvants.
  • solutions or suspensions comprising compounds according to the invention and having a pH of 4 to 10, preferably having a pH of 5 to 9, and an osmolality of 250 to 350 mosmol/kg can advantageously be prepared.
  • the pharmaceutical preparation can thus be administered directly substantially without pain intravenously, intraarterially, intraarticularly, subcutaneously or percutaneously.
  • the preparation may also be added to infusion solutions, such as, for example, glucose solution, isotonic saline solution or Ringer's solution, which may also contain further active compounds, thus also enabling relatively large amounts of active compound to be administered.
  • compositions according to the invention may also comprise mixtures of a plurality of compounds according to the invention.
  • the preparations according to the invention are physiologically well tolerated, easy to prepare, can be dispensed precisely and are preferably stable with respect to assay, decomposition products and aggregates throughout storage and transport and during multiple freezing and thawing processes. They can preferably be stored in a stable manner over a period of at least three months to two years at refrigerator temperature (2-8°C) and at rt (23-27 °C) and 60% relative atmospheric humidity (R.H.).
  • the compounds according to the invention can be stored in a stable manner by drying and when necessary converted into a ready-to-use pharmaceutical preparation by dissolution or suspension.
  • Possible drying methods are, for example, without being restricted to these examples, nitrogen-gas drying, vacuum-oven drying, lyophilisation, washing with organic solvents and subsequent air drying, liquid-bed drying, fluidised-bed drying, spray drying, roller drying, layer drying, air drying at rt and further methods.
  • the term “effective amount” denotes the amount of a medicament or of a pharmaceutical active compound which causes in a tissue, system, animal or human a biological or medical response which is sought or desired, for example, by a researcher or physician.
  • terapéuticaally effective amount denotes an amount which, compared with a corresponding subject who has not received this amount, has the following consequence: improved treatment, healing, prevention or elimination of a disease, syndrome, disease state, complaint, disorder or prevention of side effects or also a reduction in the progress of a disease, complaint or disorder.
  • therapeutically effective amount also encompasses the amounts which are effective for increasing normal physiological function.
  • the compounds according to the invention and/or physiologically acceptable salts and solvates thereof are generally used analogously to known, commercially available preparations or preparations, preferably in dosages of between 0.1 and 500 mg, in particular 5 and 300 mg, per use unit.
  • the daily dose is preferably between 0.001 and 250 mg/kg, in particular 0.01 and 100 mg/kg, of body weight.
  • the preparation can be administered one or more times per day, for example two, three or four times per day.
  • the individual dose for a patient depends on a large number of individual factors, such as, for example, on the efficacy of the particular compound used, on the age, body weight, general state of health, sex, nutrition, on the time and method of administration, on the excretion rate, on the combination with other medicaments and on the severity and duration of the particular disease.
  • a measure of the uptake of a medicament active compound in an organism is its bioavailability. If the medicament active compound is delivered to the organism intravenously in the form of an injection solution, its absolute bioavailability, i.e. the proportion of the pharmaceutical which reaches the systemic blood, i.e. the major circulation, in unchanged form, is 100%.
  • the active compound In the case of oral administration of a therapeutic active compound, the active compound is generally in the form of a solid in the formulation and must therefore first be dissolved in order that it is able to overcome the entry barriers, for example the gastrointestinal tract, the oral mucous membrane, nasal membranes or the skin, in particular the stratum corneum, or can be absorbed by the body.
  • Data on the pharmacokinetics, i.e. on the bioavailability can be obtained analogously to the method of J. Shaffer et al., J. Pharm. Sciences, 88 (1999), 313-318.
  • medicaments of this type can be prepared by means of one of the processes generally known in the pharmaceutical art.
  • Medicaments can be adapted for administration via any desired suitable route, for example by the oral (including buccal or sublingual), rectal, pulmonary, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intradermal and in particular intraarticular) routes.
  • Medicaments of this type can be prepared by means of all processes known in the pharmaceutical art by, for example, combining the active compound with the excipient(s) or adjuvant(s).
  • Parenteral administration is preferably suitable for administration of the medicaments according to the invention.
  • intra-articular administration is particularly preferred.
  • the compounds according to the invention are also suitable for the preparation of medicaments to be administered parenterally having slow, sustained and/or controlled release of active compound. They are thus also suitable for the preparation of delayed-release formulations, which are advantageous for the patient since administration is only necessary at relatively large time intervals.
  • the medicaments adapted to parenteral administration include aqueous and nonaqueous sterile injection solutions comprising antioxidants, buffers, bacteriostatics and solutes, by means of which the formulation is rendered isotonic with the blood or synovial fluid of the recipient to be treated; as well as aqueous and non-aqueous sterile suspensions, which can comprise suspension media and thickeners.
  • the formulations can be delivered in single-dose or multi-dose containers, for example sealed ampoules and vials, and stored in the freeze-dried (lyophilised) state, so that only the addition of the sterile carrier liquid, for example water for injection purposes, immediately before use is necessary.
  • Injection solutions and suspensions prepared in accordance with the formulation can be prepared from sterile powders, granules and tablets.
  • the compounds according to the invention can also be administered in the form of liposome delivery systems, such as, for example, small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles.
  • Liposomes can be formed from various phospholipids, such as, for example, cholesterol, stearylamine or phosphatidylcholines.
  • the compounds according to the invention can also be coupled to soluble polymers as targeted medicament excipients.
  • soluble polymers can encompass polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidophenol, polyhydroxyethylaspartamidophenol or polyethylene oxide polylysine, substituted by palmitoyl radicals.
  • the compounds according to the invention can furthermore be coupled to a class of biodegradable polymers which are suitable for achieving slow release of a medicament, for example polylactic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates, polylactic-co-glycolic acid, polymers, such as conjugates between dextran and methacrylates, polyphosphoesters, various polysaccharides and polyamines and poly-s-caprolactone, albumin, chitosan, collagen or modified gelatine and crosslinked or amphipathic block copolymers of hydrogels.
  • biodegradable polymers which are suitable for achieving slow release of a medicament
  • a medicament for example polylactic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates, polylactic-co-g
  • Suitable for enteral administration are, in particular, tablets, dragees, capsules, syrups, juices, drops or suppositories
  • suitable for topical use are ointments, creams, pastes, lotions, gels, sprays, foams, aerosols, solutions (for example solutions in alcohols, such as ethanol or isopropanol, acetonitrile, DMF, dimethylacetamide, 1,2-propanediol or mixtures thereof with one another and/or with water) or powders.
  • liposomal preparations are particularly suitable for topical uses.
  • the active compound in the case of formulation to give an ointment, can be employed either with a paraffinic or a water-miscible cream base. Alternatively, the active compound can be formulated to a cream with an oil-in-water cream base or a water-in-oil base.
  • Medicaments adapted to transdermal administration can be delivered as independent plasters for extended, close contact with the epidermis of the recipient.
  • the active compound can be supplied from the plaster by means of iontophoresis, as described in general terms in Pharmaceutical Research, 3 (6), 318 (1986).
  • the medicaments according to the invention may also comprise other agents usual in the art with respect to the particular type of pharmaceutical formulation.
  • the invention also relates to a set (kit) consisting of separate packs of a) an effective amount of a compound of the present invention and/or physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios, and b) an effective amount of a further medicament active compound.
  • kit consisting of separate packs of a) an effective amount of a compound of the present invention and/or physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios, and b) an effective amount of a further medicament active compound.
  • the set comprises suitable containers, such as boxes or cartons, individual bottles, bags or ampoules.
  • the set may, for example, comprise separate ampoules each containing an effective amount of a compound of the present inventionand/or pharmaceutically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios, and an effective amount of a further medicament active compound in dissolved or lyophilised form.
  • the medicaments according to the invention can be used in order to provide additive or synergistic effects in certain known therapies and/or can be used in order to restore the efficacy of certain existing therapies.
  • the pharmaceutical preparations according to the invention may also comprise further medicament active compounds, for example for use in the treatment of cancer, other anti-tumor medicaments.
  • the pharmaceutical preparations according to the invention may also, besides the compounds according to the invention, comprise further medicament active compounds which are known to the person skilled in the art in the treatment thereof.
  • methods are 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-y release, by decreasing regulatory T cell production or activation, or by increasing antigen-specific memory T cell production in a host.
  • the method comprises administering a compound of the present invention to a host in combination or alternation with an antibody.
  • the antibody is a therapeutic antibody.
  • a method of enhancing efficacy of passive antibody therapy comprising administering a compound of the present invention in combination or alternation with one or more passive antibodies.
  • This method can enhance the efficacy of antibody therapy for treatment of abnormal cell proliferative disorders such as cancer or can enhance the efficacy of therapy in the treatment or prevention of infectious diseases.
  • the compound of the present invention can be administered in combination or alternation with antibodies such as rituximab, herceptin or erbitux, for example.
  • a method of treating or preventing abnormal cell proliferation comprising administering a compound of the present invention to a host in need thereof substantially in the absence of another anticancer agent.
  • a method of treating or preventing abnormal cell proliferation in a host in need thereof comprising administering a first compound of the present invention substantially in combination with a first anticancer agent to the host and subsequently administering a second compound of the present invention receptor antagonist.
  • the second antagonist is administered substantially in the absence of another anti-cancer agent.
  • a method of treating or preventing abnormal cell proliferation in a host in need thereof comprising administering a compound of the present invention substantially in combination with a first anticancer agent to the host and subsequently administering a second anti-cancer agent in the absence of the antagonist.
  • cancer treatment disclosed here can be carried out as therapy with a compound of the present invention or in combination with an operation, irradiation or chemotherapy.
  • Chemotherapy of this type can include the use of one or more active compounds of the following categories of antitumour active compounds:
  • antiproliferative/antineoplastic/DNA-damaging active compounds and combinations thereof, as used in medical oncology such as alkylating active compounds (for example cis-platin, parboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan and nitrosoureas); antimetabolites (for example antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea and gemcitabine); antitumour antibiotics (for example anthracyclines, such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic active compounds (for example vinca alkaloids, such as vincristine, vin- bias
  • cytostatic active compounds such as anti-oestrogens (for example tamoxifen, toremifene, raloxifene, droloxifene and iodoxyfene), oestrogen receptor regulators (for example fulvestrant), anti-androgens (for example bicalutamide, flutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin), progesterones (for example megestrol acetate), aromatase inhibitors (for example anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5a-reductase, such as finasteride;
  • anti-oestrogens for example tamoxifen, toremifene, raloxifene, droloxifene and iodoxyfene
  • active compounds which inhibit cancer invasion including for example metalloproteinase inhibitors, like marimastat, and inhibitors of urokinase plasminogen activator receptor function;
  • inhibitors of growth factor function for example growth factor antibodies, growth factor receptor antibodies, for example the anti-erbb2 antibody trastuzumab [HerceptinTM] and the anti-erbbl antibody cetuximab [C225]), farnesyl transferase inhibitors, tyrosine kinase inhibitors and serine/threonine kinase inhibitors, for example inhibitors of the epidermal growth factor family (for example EGFR family tyrosine kinase inhibitors, such as 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
  • anti-angiogenic active compounds such as bevacizumab, angiostatin, endostatin, linomide, batimastat, captopril, cartilage derived inhibitor, genistein, interleukin 12, lavendustin, medroxypregesterone acetate, recombinant human platelet factor 4, tecogalan, thrombospondin, TNP-470, anti-VEGF monoclonal antibody, soluble VEGF-receptor chimaeric protein, anti-VEGF receptor antibodies, anti-PDGF receptors, inhibitors of integrins, tyrosine kinase inhibitors, serine/threonine kinase inhibitors, antisense oligonucleotides, antisense oligodexoynucleotides, siRNAs, anti-VEGF aptamers, pigment epithelium derived factor and compounds which have been published in the international patent applications WO 97/22596, WO
  • vessel-destroying agents such as combretastatin A4 and compounds which have been published in the international patent applications WO 99/02166,
  • antisense therapies for example those directed to the targets mentioned above, such as ISIS 2503, an anti-Ras antisense;
  • gene therapy approaches including, for example, approaches for replacement of abnormal, modified genes, such as abnormal p53 or abnormal BRCA1 or BRCA2, GDEPT approaches (gene-directed enzyme pro-drug therapy), such as those which use cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme, and approaches which increase the tolerance of a patient to chemotherapy or radiotherapy, such as multi-drug resistance therapy; and
  • immunotherapy approaches including, for example, ex-vivo and in-vivo approaches for increasing the immunogenicity of tumour cells of a patient, such as transfection with cytokines, such as interleukin 2, interleukin 4 or granulocyte macrophage colony stimulating factor, approaches for decreasing T-cell anergy, approaches using transfected immune cells, such as cytokine-transfected dendritic cells, approaches for use of cytokine-transfected tumour cells and approaches for use of anti-idiotypic antibodies
  • chemotherapeutic agents including foor example abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, BCG live, bevaceizumab, bexarotene, bleomycin, bortezomib, busulfan, calusterone, camptothecin, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromo
  • the medicaments from table 1 can preferably, but not exclusively, be combined with the compounds of the present invention.
  • Vss Volume of distribution (at steady state) v/v Volume to volume
  • the invention especially relates to the compounds of the following examples and physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all ratios.
  • Multiplicity is abbreviated as follows: s (singlet), d (doublet), t (triplet), q (quartet), sext (sextet), hept (heptet), m (multiplet), br (broad).
  • 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) interfaced to an Agilent 1100 mass spectrometer detector (positive mode).
  • LC-MS analyses were performed on a SHIMADZU LC-MS machine consisting of an LIFLC 20-AD system and LCMS 2020 MS detector.
  • T column: Kinetex EVO C18, 2.1x30 mm, 5.0 pm; mobile phase A: 0.0375% TFA in water (v/v), mobile phase B: 0.01875% TFA in Acetonitrile (v/v); gradient: 5% B to 95% B in 0.8 min, 95% B till min 1.2, 5% B til stop after 1.55 min; flow: 1.5 mL/min.
  • AA Analytical separation was carried out at 40°C on a Merck Chromolith Flash column (RP-18e, 25 x 2 mm) using a flow rate of 1.5 mL/min in a 2 minute gradient elution with detection at 254 nm.
  • the mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing formic acid at 0.1%.
  • 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, and then reversion back to 5:95 (A/B) over 0.05 min, finally 5:95 (A/B) for 0.2 min.
  • AH Analytical separation was carried out at 40°C on an Agilent Poroshell C18 column (30 x 2.1 mm, 2.6u, 100A) using a flow rate of 0.4 mL/min in a 4 minute gradient elution with detection at 254, 280 and 214 nm.
  • the mobile phase was a mixture of methanol (solvent A) and water (solvent B), both containing formic acid at 0.1%.
  • 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, and then reversion back to 10:90 (A/B) over 0.3 min, finally 10:90 (A/B) for 0.2 min.
  • 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 for 16 h at 100 °C. The resulting mixture was concentrated under vacuum. Then NH2OH*HCI (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.
  • A3 3-(4-methyl-1,3-oxazol-2-yl)benzoic acid
  • A3.1 To a stirred solution of [3-(methoxycarbonyl)phenyl]boronic acid (475.0 mg;
  • 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 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).
  • A5 5-Methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole
  • A5.1 lsoquinoline-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 mixed thoroughly with EtOAc (250 mL). The emulsion was then mixed with water (750 mL).
  • 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. THF was removed in vacuo, the aqueous residue was diluted with water (20 mL)and acidified to pH 3-4 with aqueous HCI-solution.
  • 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)-pyridin (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 pL, 2.826 mmol) was added and the reaction was stirred at room temperature overnight.
  • 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
  • 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-1 H-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- 5 -phosphanuide (791.0 mg, 2.819 mmol) was added at room temperature, and the mixture was stirred for 2 h.
  • Example 8.2 To a solution of intermediate 8.1 (174.0 mg, 0.394 mmol) and triethylamine (190.00 pL, 1.299 mmol) in EtOH (16.0 mL) was added Pd(dppf)Cl2.CH2Cl2 (35.1 mg, 0.043 mmol) in a pressure tank. The mixture was purged with nitrogen for 3 min and then was pressurized to 30 atm with carbon monoxide at 100°C for 32 h. The reaction mixture was cooled to room temperature and filtered to remove insoluble solids, and the filtrate was concentrated under vacuum.
  • 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
  • Example 11.1 To a stirred 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) was added diethylamine (1.45 g, 11.181 mmol) at room temperature. The resulting mixture was heated to 160 °C and stirred for 16 h. The reaction mixture was cooled to room temperature and concentrated under vacuum.
  • 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.CH2Cl2 (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 was pressurized to 30 atm with carbon monoxide at 100 °C for 16 h.
  • Example 12 Ethyl 2-(2- ⁇ [7-(1-methyl-1 H-pyrazol-4-yl)isoquinolin-1-yl]amino ⁇ ethyl)-3- oxo-2, 3-dihydro-1 H-isoindole-5-carboxylate
  • 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-1 H-isoindole-5-carboxylate
  • 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) were added triethylamine (0.76 mL, 5.18 mmol) and Pd(dppf)Cl2.CH2Cl2 (64.5 mg, 0.079 mmol) at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 2 h at 90 °C under nitrogen atmosphere. The reaction mixture was cooled to room temperature and concentrated under vacuum.
  • Example 13.2 A solution of intermediate 12.1. (87.6 mg, 0.204 mmol) in H2SO4 (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, rendered basic to pH8 with saturated aqueous NaHCOs 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 prep-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.
  • Example 15.1 To 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 pL, 2.316 mmol). The reaction mixture was heated to 45 °C while a clear solution was formed, which was stirred for 90 min at this temperature. The reaction mixture was cooled to room temperature and evaporated to dryness.
  • Example 15.2 6-methoxy-2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-2,3-dihydro-1 H-isoindol-1-one (124.0 mg, 0.293 mmol) was suspended in dry dichloromethane (4.7 mL). Boron tribromide (142.4 pL, 1.463 mmol) was dissolved in dry dichloromethane (1.9 mL), added slowly to the suspension, and the mixture was stirred at room temperature for 4 h.
  • 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 pL). Methyl 2-chloro-2,2- difluoroacetate (19.5 pL, 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 pL, 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.
  • 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
  • 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 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 down 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.
  • 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 HCI solution (0.76 mL, 32%) in a pressure tank. The mixture was hydrogenated at room temperature under 3.6 bar of hydrogen pressure over night, filtered and evaporated to dryness. The residue (824 mg, 95%) was used in the next step without further purification.
  • Example 16.3 Intermediate 16.2 (824.0 mg, 2.885 mmol) was dissolved in methanol (25.0 mL) and treated with an aqueous NaOH solution (7N, 5 mL). The reaction was stirred at room temperature for 20 min.
  • 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) while cooling in an ice bath under nitrogen atmosphere. 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 extracted with ethyl acetate. The combined extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuum.
  • sodium hydride 50% dispersion in mineral oil; 53.6 mg, 1.340 mmol
  • 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 HCI solution (64 pL, 32%) in a pressure tank. The mixture was hydrogenated at room temperature under 3.2 bar of hydrogen pressure for 17 h, filtered and evaporated to dryness. The crude product was purified by RP flash-chromatography to afford 24 mg (31%) of 2-[1-oxo-6- (trifluoromethoxy)-2,3-dihydro-1 H-isoindol-2-yl]ethan-1-ammonium formate as colourless solid. HPLC/MS m/z: 260.9 [M+H] + , Rt (G): 1.16 min.
  • Example 16.6 7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-2-ium-2olate (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 pL, 0.505 mmol) and bromo-tripyrrolidinophosphonium-hexafluorphosphate (52.2 mg, 0.112 mmol) were added and the reaction mixture was stirred at room temperature for 3 d.
  • DIPEA 85.9 pL, 0.505 mmol
  • bromo-tripyrrolidinophosphonium-hexafluorphosphate 52.2 mg, 0.112 mmol
  • 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-1 H-isoindol-1-one
  • 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 light-yellow solution was stirred at 0 °C for 10 min and then overnight at room temperature.
  • Triphenylphosphine (97.0 mg, 0.370 mmol) and di-tert-butyl azodicarboxylate (85.2 mg, 0.370 mmol) were added, the mixture was heated to 40 °C and stirred for 4.5 h.
  • the reaction mixture was diluted with dichloromethane, washed with water, dried with sodium sulfate, filtered, and evaporated to dryness.
  • the oily residue was purified by flash chromatography to yield 170 mg (94%) of methyl 5-(2,3-difluoropropoxy)-2-methylbenzoate as a light-yellow oil.
  • Example 17.2 Intermediate 16.1 (205.0 mg, 0.839 mmol), NBS (149.3 mg, 0.839 mmol) and dibenzoyl peroxide (with 25% water, 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 (with 25% water, 16.0 mg, 0.049 mmol) was added and the reaction mixture stirred at 77 °C overnight. The reaction mixture was cooled to room temperature, diluted with dichloromethane (20 mL), washed with water, dried with sodium sulfate, filtered, and evaporated to dryness.
  • Example 17.3 The reaction of intermediate B3 (48.5 mg, 0.180 mmol) and intermediate 17.2 (70.0 mg, 0.217 mmol) was performed as described for intermediate 15.1. Yield: 42 mg (49%) pale-brown solid. HPLC/MS m/z: 480.2 [M+H] + , Rt (N): 1.38 min.
  • Example 18.1 Potassium carbonate (333 mg, 2.4 mmol) was added to a mixture of 5-hydroxy-3-methyl benzoic acid methyl ester (200 mg, 1.2 mmol) and 1- iodopropane (0.24 mL, 2.4 mmol) in acetonitrile (3.3 mL) at rt. The obtained mixture was heated at reflux overnight. After 19h the reaction mixture was partitioned between ethyl acetate (60 mL) and water (40 mL).
  • Example 18.2 Methyl 2-methyl-5-propoxy-benzoate (220 mg, 1.0 mmol) was dissolved in chloroform (1.5 mL). Azobis-isobutyronitrile (AIBN) (2.6 mg, 0.14 mmol) and N-bromosuccinimide (207 mg, 1.2 mmol) were carefully added to the solution and the obtained mixture was refluxed at 65°C overnight.
  • AIBN Azobis-isobutyronitrile
  • N-bromosuccinimide 207 mg, 1.2 mmol
  • Example 18.3 A solution of methyl 2-(bromomethyl)-5-propoxy-benzoate (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 23h.
  • Example 18.4 Tert-butyl N-[2-(1-oxo-6-propoxy-isoindolin-2-yl)ethyl]carbamate (140 mg, 0.4 mmol) was mixed with 4 M HCI in dioxane (10 mL, 42 mmol) and 1 ,4- dioxane (10 mL) and stirred for 18 h. The volatiles were removed under reduced pressure and the obtained crude (176 mg) was dissolved in methanol, followed by purification using a SCX-II (2 g, 15 mL) cartridge and methanol and a 2M solution of ammonia in methanol as eluents.
  • SCX-II 2 g, 15 mL
  • Example 18.5 2-(2-Aminoethyl)-6-propoxy-isoindolin-1-one (40 mg, 0.17 mmol), 5- methyl-3-(2-oxidoisoquinolin-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 at rt under nitrogen. The reaction mixture was heated at 60°C by microwave irradiation for 1 h.
  • 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 CS2CO3 (4.47 g, 13.719 mmol) and Pd(dppf)Cl2.CH2Cl2 (0.40 g, 0.490 mmol). The resulting mixture was stirred for 16 h at 80 °C under nitrogen atmosphere.
  • Example 20.2 A stirred solution of intermediate 20.1 (1.79 g, 7.329 mmol) and diphenyl methyl sulfonium tetrafluoroborate (2.74 g, 9.516 mmol) in THF (20.0 mL) was treated with sodium bis(trimethylsilyl)amide (2M in THF, 5.86 mL, 11.711 mmol) dropwise at 0 °C under nitrogen atmosphere. The resulting mixture was warmed to room temperature and stirred for 1 h. The reaction was quenched with water and the mixture extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure.
  • 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 CCI4 (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)cyclo- propyl]benzoate as a yellow solid.
  • 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) were added PyBrOP (285.5 mg, 0.612 mmol) and DI PEA (243.5 mg, 1.884 mmol). The resulting mixture was stirred at room temperature for 2 h and concentrated under reduced pressure.
  • Example 21.1 Ethyl 5-bromo-2-methylnicotinate (1.00 g, 4.097 mmol) was brominated with NMB in CCI4 as described for 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.
  • 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.
  • Example 21.3 Carbonylation of intermediate 21.2 to the title compound was performed as described for compound 11.2. Yield: 31 mg (41 %), beige solid. HPLC/MS m/z: 444.9 [M+H] + , Rt (G): 1.28 min.
  • 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
  • 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 for 4 h at room temperature under nitrogen atmosphere and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane/methanol - 7:1) to afford 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.
  • Example 25.2 A solution of intermediate 25.1 (5.96 g, 24.222 mmol) and 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 at room temperature under nitrogen atmosphere overnight.
  • Example 25.4 To a stirred solution of intermediate 25.3 (0.88 g, 3.467 mmol) and NalC>4 (1.48 g, 6.919 mmol) in THF (10.0 mL) and water (2.0 mL) was added potassium osmate(VI) dihydrate (0.13 g, 0.345 mmol) and the mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under vacuum and the crude product (1.20 g) was used in the next step without further purification.
  • 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.
  • 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) were added CS2CO3 (542.2 mg, 1.664 mmol) and tBuXPhos Pd G3 (66.1 mg, 0.083 mmol) and the resulting mixture was stirred at 90 °C under nitrogen atmosphere for 16 h.
  • Example 25.7 Intermediate 25.5 (198.0 mg, 0.566 mmol) was dissolved in dioxane (2.0 mL), treated with a solution of HCI in dioxane (4 M, 2.0 mL) and the mixture was stirred at room temperature for 4 h. The mixture was basified to pH8 by addition of with aqueous NaHCO 3 solution. The mixture was concentrated under vacuum and the residue was purified by RP flash-chromatography to afford 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.
  • Example 25.8 Preparation 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.
  • Example 26.1 To a stirred 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) was added Ag2COs (6.77 g, 24.566 mmol), and the reaction mixture was stirred at 100 °C under nitrogen atmosphere for 2 h.
  • 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.
  • Example 26.3 To a solution of intermediate 26.2 (965.0 mg, 4.361 mmol) and NalC>4 (1.87 g, 8.743 mmol) in 1 ,4-dioxane (10.0 mL) and water (2.0 mL) was added KMnC>4 (1.03 g, 6.518 mmol), and the mixture was stirred at room temperature for 30 min. The reaction mixture was concentrated under vacuum, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate/petroleum ether - 2:3) to afford 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.
  • 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) was added sodium triacetoxyborohydride (721.6 mg, 3.405 mmol) and the mixture was stirred at room temperature for 16 h.
  • Example 26.5 Intermediate 26.4 (277.0 mg, 0.750 mmol) was stirred in an aqueous HCI solution (2N, 5.0 mL) at 50 °C for 2 h. The mixture was cooled to room temperature, neutralized to pH7 with aqueous NaHCOs solution, and concentrated under vacuum. The residue was purified by reverse flash chromatography to yield 158 mg (89%) of 2-(2-aminoethyl)-6-propoxy-1 H,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.
  • Example 26.6 Preparation 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- 1 H,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.
  • Example 27 Ethyl 1-methyl-5-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-4-oxo-1 H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate
  • 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), treated with TFA (143.74 g, 1260.612 mmol) and the resulting mixture was stirred at room temperature overnight.
  • TFA 143.74 g, 1260.612 mmol
  • 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 for 20 min at -60 °C under 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 afford 11.79 g (90%) of 2-bromo-1 H,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.
  • Example 27.3 Intermediate 27.2 (4.70 g, 21.856 mmol) was dissolved in DMF (60.0 mL) and treated with CS2CO3 (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-1 H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridin-4-one as a colorless solid.
  • 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 for 2 d at room temperature. The reaction was quenched with EtOH the mixture concentrated under reduced pressure.
  • Example 27.6 Intermediate 27.5 (184.0 mg, 0.704 mmol) was hydrogenated in EtOH (5.0 mL) in 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.
  • Example 27.7 Preparation 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-1 H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate as a colorless solid.
  • 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
  • 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
  • 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-1 H,4H,5H,6H,7H-pyrrolo[3,2-c]pyridine-2-carboxylate
  • 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) and 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 nitrogen atmosphere and stirred for 45 min. Bromoacetonitrile (69.2 mg, 5.779 mmol) was added, the reaction mixture was allowed to warm up to room temperature and stirred for 2 h. The reaction was quenched with aqueous saturated NH4CI solution and extracted with ethyl acetate.
  • 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 pl, 1.269 mmol), Pd(dppf)Cl2.CH2Cl2 (41.3 mg, 0.052 mmol) and 1 ,1-bis-(diphenylphosphino)- ferrocen (37.5 mg, 0.068 mmol) were added under nitrogen atmosphere. The reactor was pressurized to 4.0 bar with carbon monoxide and the reaction mixture stirred at 100 °C overnight.
  • Example 31.3 Hydrogenation of intermediate 31.2 (194.0 mg, 0.775 mmol) with Raney-Ni was performed 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.
  • Example 31.4 Preparation 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.
  • Example 31.5 Transesterification of intermediate 31.4 (12.0 mg, 0.026 mmol) was performed as described for 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.
  • 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.
  • Example 32.2 Intermediate 32.1 (585.0 mg, 1.792 mmol) was suspended in 1,4- dioxane (6.0 mL). A solution of HCI 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 grey-green solid. HPLC/MS m/z: 226.9 [M+H] + , Rt (H): 0.19 min.
  • Example 32.3 Under 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(ll)-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.
  • Example 32.4 Transesterification of intermediate 32.3 (18.0 mg, 0.041 mmol) was performed as described for 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.
  • Example 34.1 To a stirred solution of POCI3 (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 up 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 at 80 °C and stirred for 1 h. The reaction mixture was cooled to room temperature and concentrated under vacuum.
  • 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 under vacuum. 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.
  • 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 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.
  • Pd(PPh3)4 0.44 g, 0.381 mmol
  • 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), traces of acetic acid were 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 at room temperature for 1 h. The reaction was quenched with aqueous saturated NH4CI solution (10 mL) and the resulting mixture was extracted with dichloromethane. The combined organic layers were dried with sodium sulfate, filtered, and concentrated under reduced pressure.
  • 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 room temperature for 1 h. The reaction mixture was concentrated under vacuum 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 ⁇ -1 H-pyrrole-2-carboxylate as an off-white solid. HPLC/MS m/z: 479.25 [M+H] + , Rt (B): 0.74 min.
  • 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 under vacuum. The crude product was purified by prep-HPLC to afford 3 mg (3%) of ethyl 1-methyl-5-(2- ⁇ [7-(5-methyl-1,2,4-oxadiazol- 3-yl)isoquinolin-1-yl]amino ⁇ ethyl)-1 H,4H,5H,6H-pyrrolo[2,3-c]pyrrole-2-carboxylate as a brown oil.
  • 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.CH2Cl2 (1.52 g; 1.861 mmol) at room temperature under nitrogen atmosphere followed by traces of triethylamine. The resulting mixture was stirred at 90 °C for 3 h. The reaction mixture was cooled to room temperature and concentrated under vacuum. The residue was purified by RP flash-chromatography to give 530 mg (76%) of 5-(ethoxycarbonyl)-2-formyl-1-methyl-1 H-pyrrole-3- carboxylic acid as a yellow solid.
  • 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 for 15 min at room temperature. [Chloro(dimethylamino)methylidene]dimethylazanium; hexafluoro- 5 -phosphanuide (476.9 mg, 1.700 mmol) was added at room temperature and the mixture was stirred at room temperature for 2 h.
  • 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-1 H,4H,5H,6H-pyrrolo[2,3-c]pyrrole-2- carboxylate
  • 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
  • Example 37.1 A solution of 7-bromoisoquinolin-1-ol (1.00 g, 4.463 mmol) and tertbutyl 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.
  • 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 Pd(dppf)Cl2.CH2Cl2 (100.8 mg, 0.123 mmol) and triethylamine (373.0 mg, 3.686 mmol) were added. 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.
  • Example 37.3 Deprotection of intermediate 37.2 (475 mg, 1.318 mmol) with HCI in 1,4-dioxane was performed 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.
  • Example 37.4 A solution of intermediate 37.3 (100 mg, 0.384 mmol) and 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 dimethylsulfoxide (2.6 mL) was stirred at 120 °C for 2 d. The reaction mixture was cooled to room temperature and concentrated under reduced pressure.
  • 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
  • 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
  • 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
  • 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
  • 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
  • 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
  • Example 44.1 Ethyl 5-cyano-2-hydroxy-6-methylpyridine-3-carboxylate (10.0 g, 48.497 mmol) was stirred in POCI3 (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 was quenched by the addition of ice water (20 mL). The resulting mixture was extracted with ethyl acetate, the combined organic layers were washed with brine, dried with sodium sulfate, filtered, and concentrated under reduced pressure.
  • 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(PPh 3 ) 2 CI 2 (2.41 g, 3.434 mmol). The reaction mixture was heated to 100 °C under 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.
  • Example 44.3 A solution of intermediate 44.2 (5.92 g, 27.390 mmol) and benzyl N- (2-aminoethyl)carbamate (5.32 g; 27.390 mmol) in methanol (15.0 mL) and acetonitrile (15.0 mL) was treated with diethyl amine (7.08 g, 54.799 mmol) and irradiated in a microwave at 150 °C for 1 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure.
  • Example 44.4 Intermediate 44.3 (993.0 g, 2.724 mmol) was stirred in HCI (5.0 mL) at 100 °C for 16 h. The reaction mixture was cooled to room temperature, neutralized to pH7 with aqueous NaHCO 3 solution, and concentrated under vacuum. 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.
  • Example 44.5 Intermediate 44.4 (351.5 mg, 1.410 mmol) was dissolved in ethanol (5.0 mL), treated with H2SO4 (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 yield 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.
  • Example 44.6 Preparation 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.
  • 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
  • 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.
  • Example 46.2 Intermediate 46.1 (41.0 mg, 0.098 mmol) was suspended in dichloromethane (0.25 mL) and N-ethyldiisopropylamine (42.9 pL, 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 was stirred overnight.
  • Example 47.1 Example 46 (265.5 mg, 0.596 mmol) was dissolved in dry THF (14.8 mL) and water (7.4 mL), 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), acidified with HCI solution to pH3-4.
  • 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 pl, 1.124 mmol) was added and the reaction was stirred at room temperature overnight.
  • 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
  • Example 49.1 To a stirred solution of methyl 2,3-dihydro-1 H-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) was added triethylamine (3.99 g, 39.431 mmol) and the resulting mixture was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature and concentrated under vacuum.
  • Example 49.2 A solution of intermediate 49.1 (1.95 g, 6.086 mmol) in HCI/MeOH (20.0 mL) was stirred at room temperature for 2 h. The mixture was neutralized to pH7 with aqueous saturated NaHCOs solution (20 mL) and extracted with dichloromethane. The combined organic layers were dried with sodium sulfate, filtered, and concentrated under reduced pressure to give 1.28 g (86%) of methyl 2- (2-aminoethyl)-2,3-dihydro-1 H-isoindole-5-carboxylate as a yellow solid. HPLC/MS m/z: 221.10 [M+H] + , Rt (J): 0.29 min.
  • Example 49.3 Preparation 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-1 H-isoindole-5-carboxylate as a brown oil. HPLC/MS m/z: 430.20 [M+H] + , Rt (J): 0.53 min.
  • 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 flashchromatography to afford 75 mg (80%) of 2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3- yl)isoquinolin-1-yl]amino ⁇ ethyl)-2,3-dihydro-1 H-isoindole-5-carboxylic acid.
  • Example 49.5 Esterification was performed as described for 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-1 H-isoindole-5-carboxylate as a yellow oil.
  • 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-1 H-indazole-5-carboxylate
  • Example 51.1 7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-2-ium-2olate (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.
  • 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 down to room temperature and basified with 6N NaOH. A precipitate was 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.
  • Example 51.3 Intermediate 51.2 (448.0 mg, 1.670 mmol) was dissolved in methanol (11.0 mL) and N-methyl(tert-butoxy)carbohydrazide (293-0 mg, 2.004 mmol) and glacial acetic acid (430 pl, 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, the residue was dissolved in dichloromethane and treated with aqueous saturated NaHCCh solution. The mixture was rigorously stirred until the gas formation ceased.
  • 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. The excess of thionyl chloride was removed in vacuo, the residue was dissolved in little 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.
  • Example 51.5 Intermediate 51.4 (826.0 mg, 1.292 mmol) was dissolved in a solution of HCI in 1 ,4-dioxane (4.0 M, 11.2 mL) and stirred at room temperature for 2.5 h while a precipitate was formed. The reaction mixture was concentrated in vacuo, the residue was dissolved in dichloromethane and washed with aqueous saturated NaHCCh solution. The organic phase was dried with 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.
  • Example 51.6 Intermediate 51.5 (650.0 mg, 1.205 mmol) was dissolved in DMF (3.50 mL), 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, and the reaction mixture was heated to 90 °C and stirred for 5 h. After cooling down to room temperature, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic extracts were washed with brine, dried with sodium sulfate, filtered, and concentrated under reduced pressure.
  • Example 51.7 Saponification of intermediate 51.6 (274.0 mg, 0.598) and esterification of the corresponding acid (50 mg, 0.090 mmol) was performed as described for 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 as a colorless solid. HPLC/MS m/z: 472.8 [M+H] + , Rt (M): 1.72 min.
  • 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
  • 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
  • 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 pl, 0.551 mmol) was added, the mixture was heated to 80 °C and stirred for 18 h. The reaction mixture was cooled to room temperature, diluted with 5 mL dichloromethane and 5 ml water.
  • Example 53.2 Intermediate 53.1 (157.4 mg, 0.444 mmol) was dissolved in dry THF (7 mL) and water (4 mL). While stirring lithium hydroxide (26.6 mg, 1.110 mmol) was added and the reaction mixture was stirred at room temperature overnight. A light yellow/orange solution was formed, which was stirred at room temperature for further 48h. The reaction mixture was diluted with water (40 mL) and acidified to pH3-4 with 1.0 N HCI solution. The mixture was extracted twice with ethyl acetate, 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 NaHCCh solution and evaporated to dryness.
  • 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]-dimethyl-ammonium; hexafluoro phosphate (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.
  • HATU hexafluoro phosphate
  • N,O- dimethylhydroxylamine (21.7 mg, 0.222 mmol) and [Dimethylamino- ([1 ,2,3]triazolo[4,5-b]pyridin-3-yloxy)-methylene]-dimethyl-ammonium; hexafluoro phosphate (HATLI) (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 two times with water and saturated aqueous NaHCOs-solution (1 :1). The combined aqueous phase was extracted twice using ethyl acetate.
  • 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 in an ice bath to 0 °C. Bromo(propyl)magnesium (3M solution, 119 pl, 0.357 mmol) was added slowly and the yellow solution was allowed to warm to room temperature overnight. Further bromo(propyl)magnesium (119 pl, 0.357 mmol) was added and the reaction mixture was stirred at room temperature.
  • Example 53.5 Deprotection of intermediate 53.4 (61 mg, 0.172 mmol) with HCI in 1 ,4-dioxane was performed as described for compound 25.7 afforded 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 steps without further purification.
  • Example 54.6 Preparation as described for example 32.3. Yield: 21 mg orange solid. HPLC/MS m/z: 461.9 [M+H] + , Rt (M): 1.76 min.
  • 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
  • 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 for 30 min at 20 °C vigorously. Ethyl 3-bromo-2-oxobutanoate (1260.14 mg, 6.0282 mmol) in MeCN (6.03 mL, 0.3700 M) and iron(lll) chloride (195.56 mg, 1.2056 mmol) were added to the mixture and heating at reflux for 5 h. After completion of the reaction, as indicated by LCMS, the mixture was cooled to room temperature.
  • 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 h. Bromoacetonitrile (0.57 mL, 8.1668 mmol) was then added and the reaction was allowed to stir at room temperature overnight.
  • 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), AcOH (0.15 mL, 0.0500 M) and water (2.50 mL, 0.0500 M) was added . The reaction mixture was placed under a H2 atmosphere, and the solution was stirred at room temperature for 16 h. 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).
  • Example 54.4 5-Methyl-3-(2-oxidoisoquinolin-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 under a nitrogen atmosphere for 30 min at 40 °C in a microwave vial.
  • 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
  • Example 55.1 Methyl 6-chloro-3-methyl-pyridine-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.
  • Example 55.2 Methyl 3-(bromomethyl)-6-chloro-pyridine-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 at reflux for 1.5 h.
  • 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 backfilled with N2 (x 5, 15 sec evacuation).
  • Example 55.4 tert-Butyl /V-[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 HCI in dioxane (9.69 mL, 38.76 mmol) and anyhydrous 1,4-dioxane (10.00 mL, 0.0400 M) at room temperature under argon and stirred for overnight. Volatiles were removed under reduced pressure. The crude was dissolved in MeOH/water and filtered through a 1 g SCX column.
  • 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-oxidoisoquinolin-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) in a microwave vial at room temperature under nitrogen.
  • reaction mixture was heated at 60 °C by microwave irradiation for 1 h. Volatiles were removed under reduced pressure.
  • the crude was purified directly by reverse phase flash chromatography (10-60% MeOH in water). Fractions containing product were filtered through a 1g 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.
  • Example 56 6-(5-M ethyl- 1 ,3,4-oxadiazol-2-yl)-2-[2-[[7-(5-methyl-1 ,2,4-oxadiazol-3- yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one
  • Example 56.1 A solution of methyl 5-bromo-2-(bromomethyl)benzoate (3000.00 mg, 9.7412 mmol) and 1-Boc-ethylenediamine (1794.73 mg, 11.202 mmol), triethylamine (2.06 mL, 14.612 mmol) in MeOH (50.00 mL, 0.1900 M) was refluxed under nitrogen overnight.
  • Example 56.2 tert-Butyl N-[2-(6-bromo-1-oxo-isoindolin-2-yl)ethyl]carbamate (400.00 mg, 1.126 mmol), Xantphos Palladacycle Gen. 4 (21.70 mg, 0.0225 mmol) were mixed 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).
  • 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 HCI in dioxane (10.46 mL, 41.854 mmol) and anyhydrous 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 was dissolved in MeOH/water and filtered through a 2 g SCX column.
  • 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-oxidoisoquinolin-2-ium-7-yl)-1 ,2,4- oxadiazole [Intermediate A5] (47.51 mg, 0.2091 mmol), PyBrop (97.47 mg, 0.2091 mmol), /V,/V-diisopropylethylamine (0.11 mL, 0.6534 mmol) and anhydrous DCM (0.87 mL, 0.2000 M) in a microwave vial at room temperature under nitrogen.
  • the reaction mixture was heated at 60 °C by microwave irradiation for 2 h. Volatiles were removed under reduced pressure.
  • the crude was purified directly by reverse phase flash chromatography (10-60% MeOH in water). All fractions contained an impurity so all fractions containing product were filtered through a 2 g SCX column, the product was released with ammonia in MeOH, and the solvent evaporated in vacuo.
  • the crude mixture was then subjected to silica gel normal phase column chromatography (20-100%EtOAc:CycHex -> 10% MeOH:EtOAc).
  • Example 57.1 /V-iodosuccinimide (3493.46 mg, 15.668 mmol) was slowly added portion-wise 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 for 2 h at 0 °C and then allowed to warm to room temperature and stirred overnight.
  • Example 57.2 A solution of ethyl 4-iodo-5-methyl-1 H-pyrrole-2-carboxylate (4000.00 mg, 14.333 mmol), tert-Butyl-/V-hydroxyethyl carbamate (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 warmed to room temperature and stirred for 18 h.
  • Example 57.3 Ethyl 1-[2-(tert-butoxycarbonylamino)ethyl]-4-iodo-5-methyl-pyrrole- 2-carboxylate (2.85 g, 6.7494 mmol) was mixed with 4 N HCI in 1,4-dioxane (67.49 mL, 269.98 mmol) and anhydrous 1,4-dioxane (67.53 mL, 0.0700 M) at room temperature under argon and stirred for 2 h. Volatiles were removed under reduced pressure.
  • Example 57.4 7-lodo-6-methyl-3,4-dihydro-2H-pyrrolo[1 ,2-a]pyrazin-1-one (700.00 mg, 2.5356 mmol), Xantphos Palladacycle Gen. 4 (146.56 mg, 0.1521 mmol) were mixed in a 20 mL microwave vial.
  • the vial was capped, flushed with argon and evacuated before being fitted with a balloon filled with carbon monoxide (717.39 mg, 25.356 mmol), anyhydrous 1,4-dioxane (8.45 mL, 0.3000 M), 2-propanol (9.71 mL, 126.78 mmol) were added, and the reaction mixture was heated at 80 °C overnight.
  • 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 was added NaH (134.08 mg, 3.3521 mmol) 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 was evaporated and subjected to silica gel normal phase column chromatography (0-10% MeOH:DCM).
  • Example 57.6 To a cold solution (0 °C) 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(ll) chloride hexahydrate (259.28 mg, 1.0897 mmol) in MeOH (10.90 mL, 0.0500 M) was added NaBH4 (288.07 mg, 5.4486 mmol) (portion-wise during 10 min). The mixture was poured into hydrochloric acid (1 N, 15 mL) and stirred until the black precipitate had dissolved.
  • Example 57.7 5-Methyl-3-(2-oxidoisoquinolin-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 DI PEA (0.21 mL, 1.2082 mmol) in DCM (1.91 mL, 0.1200 M) were stirred under a nitrogen atmosphere for 30 mins at 40 °C in a microwave vial.
  • Example 58.1 1M Lithium aluminium hydride solution in THF (0.97 mL, 0.9706 mmol) was added dropwise 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) under a nitrogen atmosphere. The reaction was cooled to 0 °C and stirred for 15 min before warming room temperature and stirred for an additional 1 h. The reaction mixture was carefully quenched with 40 uL of water, followed by 40 uL of 15% NaOH aq. sol and 120 uL of water.
  • the reaction mixture was diluted with diethyl ether ( ⁇ 3 mL) and some anhydrous sodium sulfate was added. This mixture was stirred for 15 min before it was filtered off through a Telso phase separator and washed off with diethyl ether and DCM. The obtained filtrate was evaporated to dryness to give the crude product.
  • the crude has been dissolved in EtOAc (10 mL) and washed 2 times with HCI 0.5M (5 mL).
  • Example 58.2 Sodium hydride (21.24 mg, 0.5309 mmol) was added 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) under ice cooling (colourless 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 a further 4 h. The reaction mixture was partitioned between water (20 mL) and ethyl acetate (30 mL).
  • Example 58.4 2-(Ethoxymethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine hydrochloride (81.00 mg, 0.3465 mmol) was dissolved in CHCI3 (1.73 mL).
  • Triethylamine (0.05 mL, 0.3465 mmol) was added, and the reaction mixture was cooled to 0 °C.
  • a 1M 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 1h 40 min.
  • the reaction mixture was concentrated in vacuo.
  • the obtained crude (53.5 mg) was purified by silica gel column chromatography using a Biotage KP-NH 12 g column (eluent: 20-80% EtOAc in cyclohexane).
  • 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 1h 40 min. The mixture was filtered through celite, rinsed with a 1:1 mixture of DCM:EtOH.
  • 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 DI PEA (95.65 uL, 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 on at room temperature under argon overnight.
  • 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
  • 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) in a 10-20 mL microwave vial was heated under microwave irradiation for 1 h at 160 °C. A solution of saturated NaHCO 3 was added (100 mL) and the product was extracted with EtOAc (3 x 50 mL).
  • Example 59.2 A solution of methyl 2-(bromomethyl)-5-cyano-benzoate (215.00 mg, 0.8462 mmol) and /V'-[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 N2 overnight.
  • 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, mixed with EtOAc (30 mL).
  • Example 59.4 /V-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 for 10 min by microwave irradiation. Volatiles were removed under reduced pressure. The crude was directly purified by prep.
  • Example 60 6-(3-M ethyl- 1 ,2,4-oxadiazol-5-yl)-2-[2-[[7-(5-methyl-1 ,2,4-oxadiazol-3- yl)-1-isoquinolyl]amino]ethyl]isoindolin-1-one
  • 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 mixed in a microwave vial.
  • the vial was capped, evacuated before being fitted with a balloon filled with carbon monoxide. Anyhydrous 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.
  • 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 4N HCI in 1,4-dioxane (6.28 mL, 25.112 mmol) and anyhydrous 1,4-dioxane (6.00 mL) at room temperature under argon and stirred for 3 d. Volatiles were removed under reduced pressure. The crude was dissolved in MeOH/water and filtered through a 1 g SCX column.
  • 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-oxidoisoquinolin-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) in a microwave vial at room temperature under argon.
  • 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
  • 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 to the mixture was added 2-bromoacetonitrile (518 mg, 4.32 mmol, 3.00 eq). The mixture was stirred at 20 °C for 12 h.
  • 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 umol) in a mixture solvent of methanol (10 mL) and ammonium hydroxide (1 mL) was added Raney- Nickel (10.0 mg, 10% purity). The mixture was stirred at 20 °C for 12 h under hydrogen atmosphere (45 Psi).
  • 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 umol) and 7-(5-methyl- 1 ,2,4-oxadiazol-3-yl)isoquinoline 2-oxide (36.2 mg, 159 umol) in dichloromethane (5 mL) were added /V,/V-diisopropylethylamine (61.7 mg, 478 umol) and bromotri(pyrrolidin-1-yl)phosphonium hexafluorophosphate(V) (148 mg, 318 umol).
  • 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
  • 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 umol) [Example 61.3] in a mixture solvent of tetra hydrofuran (2 mL) and methanol (2 mL) was added a solution of lithium hydroxide monohydrate (32.8 mg, 782 umol) in water (1 mL). The mixture was stirred at 70 °C for 12 h.
  • 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 umol) and potassium carbonate (16.0 mg, 116 umol) in DMF (2 mL) was added 2-bromopropane (42.7 mg, 347 umol). The mixture was stirred at 80 °C for 2 h.
  • 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
  • Example 63.1 To a mixture of 2-bromo-6,7-dihydro-5/7-thieno[3,2-c]pyridin-4-one (800 mg, 3.45 mmol) in ethanol (20 mL) were added palladium acetate (77.4 mg, 345 umol), triethylamine (698 mg, 6.89 mmol), 4,5-bis(diphenylphosphino)-9,9- dimethylxanthene (598 mg, 1.03 mmol) and DMF (4 mL). The mixture was stirred at 70 °C for 12 h under carbon monoxide atmosphere (40 Psi). After the reaction was completed, the mixture was cooled to 25 °C and quenched by water (5 mL).
  • Example 63.2 To a suspension of sodium hydride (249 mg, 6.21 mmol, 60% purity) 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).
  • Example 63.3 A mixture of ethyl 5-(cyanomethyl)-4-oxo-4, 5,6,7- tetrahydrothieno[3,2-c]pyridine-2-carboxylate (250 mg, 946 umol), Raney-Nickel (81.0 mg, 94.6 umol, 10% purity) and ammonium hydroxide (1 mL) in ethanol (10 mL) was stirred at 25 °C for 12 h under hydrogen atmosphere (45 Psi). After the reaction was completed, the mixture was filtered, and the filtrate was concentrated to give a residue.
  • Example 63.4 A mixture of 7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinoline 2-oxide (50.0 mg, 220 umol), ethyl 5-(2-aminoethyl)-4-oxo-4,5,6,7-tetrahydrothieno[3,2- c]pyridine-2-carboxylate (59.0 mg, 220 umol), diisopropylethylamine (114 mg, 880 umol) and bromotri(pyrrolidin-1-yl)phosphonium hexafluorophosphate(V) (185 mg, 396 umol) in dichloromethane (5 mL) was stirred at 25 °C for 12 h.
  • Example 64 2-(2-((7-(5-Methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-7-
  • Example 65 2-(2-((7-(5-Methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-7-
  • 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 umol) [Example 62.1], 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (70.9 mg, 370 umol), 1 -hydroxybenzotriazole (37.5 mg, 278 umol) and /V,/V-diisopropylethylamine (71.7 mg, 555 umol, 96.7 uL) in acetonitrile (5 mL) was added /V-hydroxyacetamidine (27.4 mg, 370 umol).
  • Example 65.2 To a solution of /V'-((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, 123 umol) in tetrahydrofuran (3 mL) was added tetrabutylammonium fluoride (1 M in tetrahydrofuran, 368 uL, 368 umol). The mixture was stirred at 40 °C for 12 h.
  • 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-1/7-pyrrolo[3,2-c]pyridine-2-carboxylate
  • Example 66.1 To a mixture of DMF (11.4 g, 156 mmol, 12 mL) and dichloromethane (40 mL) was added phosphorus oxychloride (13.2 g, 86.3 mmol) at 0 °C slowly. 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 x 40 mL), washed with water, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give a residue.
  • Example 66.2 To a solution of benzyl 4-chloro-3-formyl-5,6-dihydropyridine-1(2/7)- 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 completed, the mixture was concentrated to give a residue.
  • Example 66.3 A mixture of benzyl 4-chloro-3-(3-ethoxy-3-oxoprop-1-en-1-yl)-5,6- dihydropyridine-1 (2/-/)-carboxylate (3.90 g, 11.15 mmol) and sodium azide (1.96 g, 30.15 mmol, 2.70 eq) 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).
  • Example 66.4 A solution of benzyl 4-azido-3-(3-ethoxy-3-oxoprop-1-en-1-yl)-5,6- dihydropyridine-1 (2/-/)-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.
  • 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-1/7-pyrrolo[3,2- c]pyridine-2,5(4/-/)-dicarboxylate (700 mg, 2.13 mmol) and iodomethane (1.56 g, 11.0 mmol, 5.16 eq). The mixture was stirred at 50 °C for 12 h. After the reaction was completed, the mixture was poured into water (5 mL).
  • Example 66.6 A mixture of 5-benzyl 2-ethyl 1-methyl-6,7-dihydro-1/7-pyrrolo[3,2- c]pyridine-2,5(4/-/)-dicarboxylate (0.7 g, 2.04 mmol) and palladium on activated carbon (0.100 g, 5% purity) in methanol (20 mL) was stirred at 25 °C for 12 h under hydrogen atmosphere (15 Psi). After the reaction was completed, the mixture was filtered. The filtrate was concentrated to afford ethyl 1-methyl-4,5,6,7-tetrahydro-1/7- pyrrolo[3,2- c]pyridine-2-carboxylate (0.22 g, crude) as a colorless oil.
  • Example 66.7 A mixture of ethyl 1-methyl-4,5,6,7-tetrahydro-1/7-pyrrolo[3,2- c]pyridine-2-carboxylate (200 mg, 960 umol), tert-butyl / ⁇ /-(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 completed, the mixture was filtered. The filtrate was concentrated to give a residue.
  • Example 66.8 A mixture of ethyl 5-(2-((tert-butoxycarbonyl)amino)ethyl)-1-methyl- 4,5,6,7-tetrahydro-1/7-pyrrolo[3,2-c] pyridine-2-carboxylate (50.0 mg, 142 umol) in hydrochloric acid/ethyl acetate (4 M, 0.5 mL) was stirred at 25 °C for 1 h. The mixture was concentrated to afford ethyl 5-(2-aminoethyl)-1-methyl-4, 5,6,7- tetrahydro-1/7-pyrrolo[3,2-c]pyridine-2-carboxylate (41.0 mg, crude, hydrochlorde) as a gray solid. HPLC/MS m/z: 252.5 [M+H] + , Rt (T): 0.22 min.
  • Example 66.9 A mixture of ethyl 5-(2-aminoethyl)-1-methyl-6,7-dihydro-4/7- pyrrolo[3,2-c]pyridine-2-carboxylate (41.0 mg, 142 umol, hydrochloride), diisopropylethylamine (73.6 mg, 570 umol), 5-methyl-3-(2-oxidoisoquinolin-2-ium-7- yl)-1 ,2,4-oxadiazole [Intermediate A5] (32.4 mg, 142 umol) and bromo(tripyrrolidin-1- yl)phosphonium;hexafluorophosphate (120 mg, 256 umol) in DMF (2.50 mL) was stirred at 25 °C for 12 h.
  • Example 67.1 To a solution of 7-bromo-3,4-dihydroisoquinolin-1(2/7)-one (1.50 g, 6.64 mmol), triethylamine (1.34 g, 13.3 mmol) in a mixture solvent of ethanol (20 mL) and DMF (4 mL) were added palladium(ll) acetate (149 mg, 664 umol) and (5- diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenyl-phosphane (1.15 g, 1.99 mmol). The mixture was stirred at 70 °C for 24 h under carbon monoxide atmosphere (45 Psi).
  • 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 to the mixture was added 2-bromoacetonitrile (1 .31 g, 11.0 mmol) was added. The mixture was stirred at 20 °C for 12 h.
  • 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 mixture solvent of methanol (20 mL) and ammonium hydroxide (4 mL) was added Raney-nickel (60.0 mg, 10% purity). The mixture was stirred at 20 °C for 12 h under hydrogen atmosphere (45 Psi). The mixture was filtered and concentrated to give a residue.
  • Example 67.4 To a solution of ethyl 2-(2-aminoethyl)-1-oxo-1 , 2,3,4- tetrahydroisoquinoline-7-carboxylate (60.0 mg, 201 umol, hydrochloride), N,N- diisopropylethylamine (123 mg, 953 umol) in dichloromethane (5 mL) were added 5- methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (43.3 mg, 191 umol) and bromotripyrrolidinophosphonium hexafluorophosphate (178 mg, 381 umol).
  • 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
  • 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-oxo-propanoate (33.6 g, 172 mmol) in acetonitrile (60 mL) and iron(lll) chloride (5.59 g, 34.5 mmol) at 0 °C. The mixture was stirred at 80 °C for 12 h.
  • 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 borontrifluoridediethyletherate (46.0 g, 324 mmol, 40.0 mL) in tetrahydrofuran (300 mL) was added sodium borohydride (8.00 g, 211 mmol) in portions 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 hydrochloric acid aqueous solution (10% 30 mL) and the solution was stirred for 2 h.
  • the resulting solution was neutralized with saturated sodium bicarbonate aqueous solution and 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 x 200 mL). The combined organic layers were concentrated to give a residue.
  • 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 / ⁇ /-(2- bromoethyl)carbamate (1.15 g, 5.13 mmol) in DMF (5 mL) was added N,N- diisopropylethylamine (798 mg, 6.18 mmol) and potassiumiodide (34.2 mg, 206 umol). The mixture was stirred at 90 °C for 12 h. The mixture was concentrated to give a residue.
  • Example 68.4 A solution of ethyl 2-(2-((terf-butoxycarbonyl)amino)ethyl)-1 , 2,3,4- tetrahydropyrrolo[1 ,2-a]pyrazine-7-carboxylate (0.350 g, 1.04 mmol) in hydrochloric acid/ethyl acetate (4 M, 5 mL) was stirred at 20 °C for 2 h.
  • 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 umol, hydrochloride), 5-methyl-3-(2- oxidoisoquinolin-2-ium-7-yl)-1 ,2,4-oxadiazole [Intermediate A5] (125 mg, 548 umol) and /V,/V-diisopropylethylamine (354 mg, 2.74 mmol) in dichloromethane (2 mL) was added bromotripyrrolidinophosphonium hexafluorophosphate (511 mg, 1.10 mmol).
  • 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 umol) in a mixture solvent of tetrahydrofuran (2 mL) and methanol (2 mL) was added a solution of lithium hydroxide monohydrate (12.7 mg, 302 umol) in water (1 mL). The mixture was stirred at 70 °C for 12 h.
  • 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 umol) 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 prep-HPLC (column: Phenomenex Gemini-NX C18 75*30mm*3um; mobile phase: [water(10mM NH4HCO3)-ACN]; B%: 40%-70%, 8 min) and lyophilized to afford 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 (15.64 mg, 33.6 umol, 28% yield, 99% purity) as a yellow solid.
  • 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.
  • Example 69.2 06-tert-butyl 02-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, 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, stirred for another 6 h.
  • 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 uL, 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 MgSO4 and concentrated in vacuo.
  • 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.
  • 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
  • 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 dropwise addition of boron trifluoride diethyl etherate (0.11 mL, 0.8823 mmol). The reaction was warmed to room temperature for 1 h.
  • 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 ditert-butyl 5,7-dihydro-4H-thieno[2,3-c]pyridine-2,6-dicarboxylate in an analogous manner shown in Example 69.
  • 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 uL, 0.4599 mmol) was added and the reaction mixture was stirred at room temperature for 2 d.
  • DIPEA 80.11 uL, 0.4599 mmol
  • 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
  • Example 72.1 4,5,6,7-Tetrahydropyrazolo[1 ,5-a]pyrazine-2-carboxylic acid hydrochloride (600.00 mg, 2.9465 mmol) was charged 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.
  • Example 72.2 05-tert-butyl 02-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, 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 stirred for 16 h.
  • 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 CHCI3 (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, stirred at 0 °C for 50 min. The mixture was concentrated in vacuo.
  • 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, the atmosphere was replaced by hydrogen. The reaction mixture was stirred at room temperature for 22 h. The reaction mixture was filtered through celite, rinsed with EtOH. The solvent was removed under reduced pressure.
  • 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.
  • Example 73.1 Propan-2-yl 5-(2-aminoethyl)-6,7-dihydro-4H-pyrazolo[1,5- a]pyrazine-2-carboxylate was prepared by an analogous procedure to Example 72.
  • 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, stirred for another 1 h.
  • Example 73.3 Propan-2-yl 5-(2-aminoethyl)-6,7-dihydro-4H-pyrazolo[1 ,5- a]pyrazine-2-carboxylate (30.00 mg, 0.1189 mmol) used in an analogous procedure to Example 72.5 to afford 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.
  • 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
  • Example 75.1 2-[2-[[7-(5-Methyl-1 ,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3- oxo-isoindoline-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, mixed with EtOAc (30 mL).
  • Example 76 6-(5-Ethyl- 1 ,2,4-oxadiazol-3-yl)-2-[2-[[7-(5-methyl-1 ,2,4-oxadiazol-3-yl)-
  • 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) in a 10-20 mL microwave vial was heated under microwave irradiation for 1 h at 160 °C. A solution of saturated bicarbonate was added (100 mL) and the product was extracted with EtOAc (3 x 50 mL).
  • 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.
  • 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 mixed in a microwave vial. The capped vial was evacuated before fitted with a balloon filled with carbon monoxide.
  • Example 78.1 2-Chloro-5-methyl-isonicotinic 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.
  • 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 at reflux for 30 min. The reaction mixture was cooled to ambient temperature.
  • 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)Ch ⁇ DCM (41.32 mg, 0.0481 mmol) were mixed in a microwave vial. The vial was capped and evacuated before being fitted with a balloon filled with carbon monoxide.
  • 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 HCI in 1,4- dioxane (5.22 mL, 20.894 mmol) and anyhydrous 1 ,4-dioxane (5.00 mL) at room temperature under argon and stirred for 16 h. Volatiles were removed under reduced pressure. The crude was dissolved in MeOH and filtered through a 1 g SCX-2 column.
  • 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 at room temperature under argon. The reaction mixture was heated at 60 °C by microwave irradiation for 1 h.
  • Example 79 2-[2-[[7-(5-Methyl-1 ,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6- (propylamino)isoindolin-l-one
  • 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.
  • Example 79.2 Analogous procedures to those used in Example 78.4 and Example 78.5 afforded 2-[2-[[7-(5-methyl-1 ,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6- (propylamino)isoindolin-l-one (29 mg, 0.0633 mmol) as an off-white solid.
  • Example 80.2 Methyl 3-(bromomethyl)-6-chloro-pyridine-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 at reflux for 1.5 h. The reaction mixture was cooled to ambient temperature.
  • 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 mixed in a microwave vial. The vial was capped, flushed with argon and evacuated before being fitted with a balloon filled with carbon monoxide.
  • 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 HCI in 1 ,4- dioxane (1.47 mL, 5.8675 mmol) and anyhydrous 1,4-dioxane (3.00 mL) at room temperature under argon and stirred for 2 h. Volatiles were removed under reduced pressure. The crude was dissolved in MeOH and filtered through a 1 g SCX-2 column.
  • 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-oxidoisoquinolin-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) in a microwave vial at room temperature under argon. The reaction mixture was heated at 60 °C by microwave irradiation for 1 h.
  • Example 82 2-[2-[[7-(5-Methyl-1 ,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6- nitro-isoindolin-1-one
  • Example 83 N-[2-[[7-(5-Methyl-1 ,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-3- oxo-isoindolin-5-yl]propanamide 2-[2-[[7-(5-methyl-1,2,4-oxadiazol-3-yl)-1-isoquinolyl]amino]ethyl]-6-nitro-isoindolin- 1-one [Example 82] (50.00 mg, 0.1141 mmol) was dissolved in anhydrous DMF (0.30 mL) under argon.
  • Zinc powder 29.83 mg, 0.4563 mmol
  • glacial acetic acid 0.05 mL, 0.9126 mmol
  • Triethylamine (0.06 mL, 0.4563 mmol)
  • propionyl chloride (0.01 mL, 0.1255 mmol) were added and the reaction mixture was continued to stir for 1 h. Volatiles were removed under reduced pressure.
  • Example 84 Ethyl 6-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl)amino)ethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-carboxylate
  • Example 84.1 To a solution of ethyl 4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2- carboxylate hydrochloride (100 mg, 0.4 mmol) in anhydrous DMF (1.4 mL) was added N-Boc-2-chloroethylamine (181.3 mg, 1.0 mmol), potassium carbonate (139.5 mg, 1.0 mmol) and potassium iodide (2.7 mg, 0.02 mmol). The reaction mixture was stirred at 70 °C overnight. The reaction progress was followed by LC-MS and after 19 h showed nearly reaction completion. The reaction mixture was diluted with ethyl acetate (90 mL) and water (30 mL).
  • Example 84.2 Trifluoroacetic acid (0.5 mL, 6.4 mmol) was added drop wise to a solution of ethyl 6-[2-(tert-butoxycarbonylamino)ethyl]-5,7-dihydro-4H-thieno[2,3- c]pyridine-2-carboxylate (56.4 mg, 0.16 mmol) in dichloromethane (0.5 mL) and this was stirred for 2 h 15 min. The volatiles were then evaporated to dryness. The obtained crude was purified by SCX-2 cartridge (2 g, 15 mL) using methanol and 2M ammonia solution in methanol as eluents.
  • Example 84.3 6-(2-Aminoethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate (33.6 mg, 0.13 mmol) was dissolved in anhydrous dichloromethane (1.32 mL), followed by addition of N,N-diisopropylethylamine (86 uL, 0.50 mmol) followed by 5- methyl-3-(2-oxidoisoquinolin-2-ium-7-yl)-1,2,4-oxadiazole [Intermediate A5] (30 mg, 0.13 mmol) and PyBroP (80 mg, 0.17 mmol). After 16 h, LC-MS showed good conversion to the desired product.
  • the crude was purified by flash column chromatography (eluent: gradient of 10-100% ethyl acetate in cyclohexane) to give ethyl 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 that wasn't pure.
  • the product was further purified by reverse phase column (eluent: 30-100% of MeOH in water both modified with 0.1% formic acid).
  • Example 85 Methyl 6-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl)amino)ethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-carboxylate
  • Example 85.1 Triethylamine (71 uL mL, 0.51 mmol) was added to a solution of methyl 4H,5H,6H,7H-thieno[2,3-c]pyridine-2-carboxylate hydrochloride (125 mg, 0.51 mmol) in anhydrous chloroform (2.53 mL). The reaction mixture was cooled to 0 °C and a 1M solution of 1-nitroethene in xylene (0.51 mL, 0.51 mmol) was added. The reaction mixture was stirred at 0 °C for 10 min and then warmed up to room temperature.
  • Example 85.2 Zinc (185.6 mg, 2.83 mmol), ammonium chloride (151.8 mg, 2.83 mmol) and methyl 6-(2-nitroethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-2-carboxylate (95.9 mg, 0.35 mmol) were suspended in a mixture of tetrahydrofuran (1.4 mL) / water (0.45 mL) / MeOH (0.5 mL). The obtained reaction mixture was stirred at room temperature. After 2 h, the reaction mixture was filtered on celite and rinsed with a 1 :1 mixture of dichloromethane:methanol (20 mL).
  • Example 85.3 Methyl 6-(2-aminoethyl)-5,7-dihydro-4H-thieno[2,3-c]pyridine-2- carboxylate (68.8 mg, 0.29 mmol) was dissolved in anhydrous dichloromethane (2.9 mL) followed by addition of N,N-diisopropylethylamine (187 uL, 1.1 mmol), 5-methyl- 3-(2-oxidoisoquinolin-2-ium-7-yl)-1 ,2,4-oxadiazole [Intermediate A5] (65 mg, 0.29 mmol) and PyBroP (173 mg, 0.37 mmol).
  • Example 86 Methyl 6-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl)amino)ethyl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine-2-carboxylate
  • Example 87 Ethyl 1-methyl-2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-1 H-1 ,3-benzodiazole-5-carboxylate
  • Example 87.1 A solution of ethyl 3-amino-4-(methylamino)benzoate (300.0 mg, 1.545 mmol), 3-[[7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino]propanoic acid (307.0 mg, 1.027 mmol) and NMI (190.0 mg, 2.314 mmol) in DMF (5.0 mL) was stirred for 5 min at room temperature.
  • Example 87.2 A solution of ethyl 3-(3-[[7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin- 1-yl]amino]propanamido)-4-(methylamino)benzoate (85.5 mg, 0.180 mmol) in AcOH (3.0 mL) was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature, diluted with brine (3 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layers were dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure.
  • Example 88 Propan-2-yl 1-methyl-2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3- yl)isoquinolin-1-yl]amino ⁇ ethyl)-1 H-1 ,3-benzodiazole-5-carboxylate
  • Example 90 Ethyl 2-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl)amino)ethyl)-3H-imidazo[4,5-b]pyridine-6-carboxylate
  • Example 90.1 Preparation as described for Example 87.1 using methyl 5,6- diaminopyridine-3-carboxylate (659.0 mg, 3.942 mmol) and 3- ⁇ [7-(5-methyl-1 ,2,4- oxadiazol-3-yl)isoquinolin-1-yl]amino ⁇ propanoic acid (1.39 g, 4.643 mmol), NMI (2.42 g, 29.507 mmol) and TCFH (2.28 g, 8.126 mmol) in DMF (20.0 mL).
  • Example 90.2 Cyclisation as described for Example 87.2. Yield: 108 mg (44%) of methyl 2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino ⁇ ethyl)-3H- imidazo[4,5-b]pyridine-6-carboxylate as a black solid. HPLC/MS m/z: 430.10 [M+H] + , Rt (B): 0.72 min.
  • Example 90.3 Saponification with lithium hydroxide in THF and water yielded 50 mg of 2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino ⁇ ethyl)-3H- imidazo[4,5-b]pyridine-6-carboxylic acid as a light-brown solid.
  • Example 90.4 Esterification with H2SO4 in ethanol provided 5 mg of ethyl 2-(2-((7- (5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl)amino)ethyl)-3H-imidazo[4,5- b]pyridine-6-carboxylate as a colorless solid.
  • Example 92 tert-Butyl 2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-3H-imidazo[4,5-b]pyridine-6-carboxylate 5 mg light-brown solid.
  • Example 96.1 To a stirred solution of 5-bromo-6-methylpyridine-2,3-diamine (1.57 g, 7.770 mmol) and 3- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ propanoic acid (2.17 g, 7.275 mmol) in DMF (20.0 mL) were added 1- methyl-1 H-imidazole (1.52 g, 18.513 mmol) and TCFH (3.04 g, 10.835 mmol). The resulting mixture was stirred at room temperature for 16 h.
  • Example 96.2 A mixture of N-(2-amino-5-bromo-6-methylpyridin-3-yl)-3-[[7-(5- methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino]propanamide (180.0 mg, 0.373 mmol) in AcOH (2.0 mL) was stirred at 150 °C for 20 min. The resulting mixture was codes to room temperature and concentrated under vacuum.
  • Example 96.3 Carbonylation using the described conditions yielded 8 mg (7%) of ethyl 5-methyl-2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino ⁇ ethyl)- 3H-imidazo[4,5-b]pyridine-6-carboxylate as a colorless solid.
  • 1 H NMR (300 MHz, DMSO-d 6 ): 5 12.93 (s, 1 H), 8.85 (d, J
  • Example 98 Propan-2-yl 3,5-dimethyl-2-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3- yl)isoquinolin-1-yl)amino)ethyl)-3H-imidazo[4,5-b]pyridine-6-carboxylate 21 mg colorless solid. HPLC/MS m/z: 486.15 [M+H] + , Rt (B): 0.78 min.
  • Example 100 Ethyl 6-methyl-2-(2- ⁇ [7-(5-methyl-1,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-1H-imidazo[4,5-b]pyridine-5-carboxylate
  • Example 102 Ethyl 2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-1,3-benzothiazole-5-carboxylate
  • Example 102.1 A solution of 2-amino-4-bromobenzenethiol (117.0 mg, 0.573 mmol) and 3-[[7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino]propanenitrile (160.0 mg, 0.573 mmol) in AcOH (3.0 mL) and EtOH (3.00 mL) was stirred at 80 °C under nitrogen atmosphere overnight. The resulting mixture was cooled to room temperature and concentrated under reduced pressure. The residue was diluted with water (5 mL). The aqueous layer was extracted with EtOAc (3 x 10 mL). The combined organic layers were dried with sodium sulfate, filtered, and concentrated in vacuum.
  • Example 102.2 Carbonylation using the described conditions yielded 21 mg (11%) of ethyl 2-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1-yl]amino ⁇ ethyl)-1 ,3- benzothiazole-5-carboxylate as an off-white solid.
  • Example 103 Ethyl 7-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-8-oxo-1 ,2,3,4,7,8-hexahydro-2,7-naphthyridine-2-carboxylate
  • Example 103.1 2H-[2,7]Naphthyridin-1-one (500.0 mg, 3.421 mmol), 2-(Boc- amino)ethyl bromide (920.0 mg, 4.105 mmol) and potassium carbonate (1.42 g, 10.264 mmol) were dissolved in dry DMF (13.3 mL). The suspension was heated to 80 °C and stirred for 4 h. After 1-2 h, the reaction mixture turned red.
  • Example 103.2 tert-Butyl N-[2-(1-oxo-1 ,2-dihydro-2,7-naphthyridin-2- yl)ethyl]carbamate (691.0 mg, 2.388 mmol) were hydrogenated in THF (10 mL) and acetic acid (2 mL) in presence of PtC ⁇ xFLO (130 mg) at room temperature for 6 h at 1 bar. The catalyst was filtered off and the filtrate was evaporated. The residue was used in the next step without further purification.
  • Example 103.3 To a solution of tert-butyl N-[2-(1-oxo-1 ,2,5,6,7,8-hexahydro-2,7- naphthyridin-2-yl)ethyl]carbamate (250.0 mg, 0.579 mmol) and dry pyridine (468 pl, 5.795 mmol) in dry dichloromethane (3.7 mL) was added ethyl chloroformate (61 pl, 0.637 mmol).
  • Example 103.4 Ethyl 7-(2- ⁇ [(tert-butoxy)carbonyl]amino ⁇ ethyl)-8-oxo-1 ,2,3,4,7,8- hexahydro-2,7-naphthyridine-2-carboxylate (79.0 mg, 0.216 mmol) was dissolved in a solution of HCI in 1 ,4-dioxane (4.0 M, 3.8 mL) and stirred 15 min at room temperature. All volatiles were removed in vacuum and the resulting oil (75 mg) was used without further purification.
  • Example 103.5 7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-2-ium-2olate (20.0 mg, 0.088 mmol) and 2-[7-(ethoxycarbonyl)-1-oxo-1 ,2,5,6,7,8-hexahydro-2,7- naphthyridin-2-yl]ethan-1-amine hydrochloride (33.2 mg, 0.110 mmol) were dissolved in dry dichloromethane (1 mL) under an argon atmosphere.
  • N- Ethyldiisopropylamine for synthesis 101 pL, 0.594 mmol
  • bromo- tripyrrolidinophosphonium-hexafluorphosphate 53.1 mg, 0.114 mmol
  • Example 104 Propyl 7-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-8-oxo-1 ,2,3,4,7,8-hexahydro-2,7-naphthyridine-2-carboxylate
  • Example 105 Propan-2-yl 7-(2- ⁇ [7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl]amino ⁇ ethyl)-8-oxo-1 ,2,3,4,7,8-hexahydro-2,7-naphthyridine-2-carboxylate
  • Example 106 Propan-2-yl 2-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl)amino)ethyl)-1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate
  • Example 106.1 To a solution of 2-bromopropane (865 mg, 7.03 mmol, 1.50 eq) in dimethyl formamide (10 mL) was added potassium carbonate (1.30 g, 9.38 mmol, 2.00 eq). The mixture was stirred at 60 °C for 0.5 h. To the mixture was added 2- (terf-butoxycarbonyl)-1,2,3,4-tetrahydroisoquinoline-7-carboxylic acid (1.30 g, 4.69 mmol, 1.00 eq). The mixture was stirred at 60 °C for 3 h.
  • Example 106.2 A solution of 2-terf-butyl 7-isopropyl 3,4-dihydroisoquinoline- 2,7(1 /-/)-dicarboxylate (0.600 g, 1.95 mmol, 1.00 eq) in hydrochloric acid/ethyl acetate (4 M, 10 mL) was stirred at 20 °C for 2 h. The mixture was concentrated under reduced pressure to afford isopropyl 1,2,3,4-tetrahydroisoquinoline-7- carboxylate (0.450 g, crude, hydrochloride) as a brown solid.
  • Example 106.3 To a solution of isopropyl 1 ,2,3,4-tetrahydroisoquinoline-7- carboxylate (1.00 g, 3.91 mmol, 1.00 eq, hydrochloride) and N,N- diisopropylethylamine (2.53 g, 19.6 mmol, 5.00 eq) in dimethyl formamide (10 mL) were added potassium iodide (649 mg, 3.91 mmol, 1.00 eq) and tert-butyl / ⁇ /-(2- bromoethyl)carbamate (2.63 g, 11.7 mmol, 3.00 eq). The mixture was stirred at 90 °C for 12 h.
  • Example 106.4 A solution of ethyl 5-(2-((tert-butoxycarbonyl)amino)ethyl)-1-methyl- 4,5,6,7-tetrahydro-1/7-pyrrolo[3,2- c]pyridine-3-carboxylate (0.250 g, 711 umol, 1.00 eq) in hydrochloric acid/ethyl acetate (4 M, 5 mL) was stirred at 20 °C for 2 h. The mixture was concentrated under reduced pressure to afford isopropyl 2-(2- aminoethyl)-1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate (0.200 g, crude, hydrochloride) as a yellow solid.
  • Example 106.5 To a solution of isopropyl 2-(2-aminoethyl)-1 , 2,3,4- tetrahydroisoquinoline-7-carboxylate (65.8 mg, 220 umol, 1.25 eq, hydrochloride), /V,/V-diisopropylethylamine (114 mg, 880 umol, 5.00 eq) in dichloromethane (5 mL) were added 7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinoline 2-oxide (40.0 mg, 176 umol, 1.00 eq) and bromotripyrrolidinophosphonium hexafluorophosphate (164 mg, 352 umol, 2.00 eq) at 20 °C.
  • Example 107 Ethyl 2-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl)amino)ethyl)-1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate
  • Example 107.1 A mixture of 2-terf-butoxycarbonyl-3,4-dihydro-1/7-isoquinoline-7- carboxylic acid (900 mg, 3.25 mmol, 1.00 eq), iodoethane (607.
  • Example 107.2 A mixture of 2-terf-butyl 7-ethyl 3,4-dihydroisoquinoline-2,7(1/7)- dicarboxylate (300 mg, 982 umol, 1.00 eq) in hydrochloric acid/ethyl acetate (4 M, 10 mL) was stirred at 25 °C for 0.5 h. After the reaction was completed, the mixture was concentrated to afford ethyl 1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate (230 mg, crude, hydrochloride) as a white solid.
  • Example 107.3 A mixture of ethyl 1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate (230 mg, 951 umol, 1.00 eq, hydrochloride), cesium carbonate (775 mg, 2.38 mmol, 2.50 eq) and terf-butyl / ⁇ /-(2-bromoethyl)carbamate (235 mg, 1.05 mmol, 1.10 eq) in acetonitrile (5 mL) was stirred at 60 °C for 12 h. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated to give a residue.
  • Example 107.4 A mixture of ethyl 2-(2-((terf-butoxycarbonyl)amino)ethyl)-1 , 2,3,4- tetrahydroisoquinoline-7-carboxylate (260 m g, 746umol, 1.00 eq) in hydrochloric acid/ethyl acetate (4 M, 10 mL) was stirred at 25 °C for 1 h. After the reaction was completed, the mixture was concentrated to afford ethyl 2-(2-aminoethyl)-1 , 2,3,4- tetrahydroisoquinoline-7-carboxylate (220 mg, crude, hydrochloride) as a yellow solid.
  • Example 107.5 A mixture of ethyl 2-(2-aminoethyl)-1 ,2,3,4-tetrahydroisoquinoline-7- carboxylate (100 mg, 351 umol, 1.00 eq, hydrochloric acid salt), 5-methyl-3-(2- oxidoisoquinolin-2-ium-7-yl)-1 ,2,4-oxadiazole (79.7 mg, 351umol, 1 eq), diisopropylethylamine (181 mg, 1.40 mmol, 4.00 eq) and bromo(tripyrrolidin-1- yl)phosphonium; hexafluorophosphate (295 mg, 632 umol, 1.80 eq) in dichloromethane (5 mL) was stirred at 25 °C for 12 h.
  • Example 108 Methyl 2-(2-((7-(5-methyl-1 ,2,4-oxadiazol-3-yl)isoquinolin-1- yl)amino)ethyl)-1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate
  • Example 108.1 Methyl 1 ,2,3,4-tetrahydroisoquinoline-7-carboxylate HCL (100.00 mg, 0.4392 mmol) , N-Boc-2-chloroethylamine (201.27 mg, 1.098 mmol), Potassium carbonate (212.44 mg, 1.5372 mmol) and potassium iodide (7.29 mg, 0.0439 mmol) were dissolved in dry DMF (2.20 mL, 0.2000 M ). The reaction mixture was stirred at 70 °C for 16 h.
  • Example 108.2 Methyl 2-[2-(tert-butoxycarbonylamino)ethyl]-3,4-dihydro-1 H- isoquinoline-7-carboxylate (80.00 mg, 0.2392 mmol) was dissolved in dry 1 ,4- dioxane (1.20 mL, 0.2000 M). 4N hydrogen chloride in dioxane (0.60 mL, 2.3923 mmol) was added and the reaction mixture was stirred overnight at room temperature.
  • Example 109 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

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Abstract

L'invention concerne des bicycles substitués de formule générale I, et l'utilisation des composés de la présente invention pour le traitement et/ou la prévention de maladies et de troubles hyperprolifératifs chez des mammifères, en particulier des êtres humains, et des compositions pharmaceutiques contenant un tel composé.
PCT/EP2023/072876 2022-08-23 2023-08-21 Bicycles substitués utilisés comme inhibiteurs de hset Ceased WO2024042007A1 (fr)

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CN202380074493.0A CN120091998A (zh) 2022-08-23 2023-08-21 作为hset抑制剂的取代的双环
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JP2025511776A JP2025529897A (ja) 2022-08-23 2023-08-21 Ｈｓｅｔインヒビターとしての置換二環系
EP23757923.0A EP4577537A1 (fr) 2022-08-23 2023-08-21 Bicycles substitués utilisés comme inhibiteurs de hset
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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006031348A2 (fr) * 2004-08-18 2006-03-23 Merck & Co., Inc. Inhibiteurs de la kinesine mitotique
WO2007056469A2 (fr) * 2005-11-09 2007-05-18 Cytokinetics, Inc. Entites et compositions chimiques et methodes associees
WO2023131690A1 (fr) * 2022-01-10 2023-07-13 Merck Patent Gmbh Hétérocycles substitués en tant qu'inhibiteurs de hset

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006031348A2 (fr) * 2004-08-18 2006-03-23 Merck & Co., Inc. Inhibiteurs de la kinesine mitotique
WO2007056469A2 (fr) * 2005-11-09 2007-05-18 Cytokinetics, Inc. Entites et compositions chimiques et methodes associees
WO2023131690A1 (fr) * 2022-01-10 2023-07-13 Merck Patent Gmbh Hétérocycles substitués en tant qu'inhibiteurs de hset

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EP4577537A1 (fr) 2025-07-02
AR130226A1 (es) 2024-11-20
AU2023328866A1 (en) 2025-04-03
CA3265564A1 (fr) 2024-02-29

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