HK1205123B

HK1205123B - Disubstituted benzothienyl-pyrrolotriazines and their use as fgfr kinase inhibitors

Info

Publication number: HK1205123B
Application number: HK15105900.1A
Authority: HK
Inventors: Dirk Brohm; Melanie HEROULT; Marie-Pierre Collin; Walter Hübsch; Mario Lobell; Klemens Lustig; Sylvia Grünewald; Ulf Bömer; Verena Voehringer
Original assignee: Bayer Pharma Aktiengesellschaft; Bayer Intellectual Property Gmbh
Priority date: 2011-12-15
Filing date: 2012-12-10
Publication date: 2018-02-02

Description

Disubstituted benzothienyl-pyrrolotriazines and their use as FGFR kinase inhibitors

The present invention relates to novel substituted 5- (1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine derivatives having protein tyrosine kinase inhibitory activity, to processes for preparing such compounds, to pharmaceutical compositions containing such compounds, and to the use of such compounds or compositions for the treatment of proliferative diseases, in particular cancer and tumor diseases.

Cancer is the leading cause of global death, resulting in the death of 760 million people in 2008 (about 13% of all deaths). In 2030, the number of deaths due to cancer is expected to continue to rise worldwide to over 1 thousand 1 million (WHO source, fatsheet number 297, 2 months 2011).

There are many ways how cancers arise, which is one of the reasons why their treatment is difficult. One way in which cell transformation can occur is to follow genetic changes. The completion of the human genome project revealed genomic instability and heterogeneity of human oncogenes. Recent strategies to identify these genetic changes have accelerated the process of oncogene discovery. Genetic abnormalities may, for example, lead to overexpression of proteins and thus to a non-physiological activation of these proteins. One family of proteins from which many oncoproteins are derived are tyrosine kinases, particularly Receptor Tyrosine Kinases (RTKs). Over the last two decades, a number of research approaches have suggested the importance of RTK-mediated signaling in the poor cell growth leading to cancer. In recent years, promising results have been achieved in the clinic using selective small molecule inhibitors of tyrosine kinases as a new class of anti-tumorigenic agents [ Swinney and Anthony,Nature Rev. Drug Disc.10 (7), 507-519 (2011)]。

fibroblast Growth Factors (FGFs) and their receptors (FGFRs) form part of a unique and distinct signaling system,it plays a key role in various biological processes (which encompass various aspects of embryonic development and adult pathophysiology) [ Itoh and Ornitz,J. Biochem.149 (2), 121-130 (2011)]. In a spatiotemporal manner, FGFs stimulate a wide range of cellular functions, including migration, proliferation, differentiation and survival, through FGFR binding.

The FGF family contains 18 secreted polypeptide growth factors that bind to four highly conserved receptor tyrosine kinases (FGFR-1 to-4) expressed on the cell surface. Furthermore, FGFR-5 can bind FGF, but does not have a kinase domain, and thus lacks intracellular signaling. The specificity of the ligand/receptor interaction is enhanced by a number of transcriptional and translational processes, which lead to multiple isoforms through alternative transcriptional initiation, alternative splicing, and C-terminal truncation. Various heparan sulfate proteoglycans (e.g., syndecans) may be part of the FGF/FGFR complex and strongly influence the ability of FGFs to induce signaling responses [ Polanska et al,Developmental Dynamics238 (2), 277-293 (2009)]. FGFR is a cell surface receptor consisting of three extracellular immunoglobulin-like domains, a single-pass transmembrane domain, and an intracellular dimerization tyrosine kinase domain. Binding of FGFs brings the intracellular kinases into close proximity, enabling them to transphosphorylate each other. Seven phosphorylation sites have been identified (e.g., Tyr463, Tyr583, Tyr585, Tyr653, Tyr654, Tyr730, and Tyr766 in FGFR-1).

Some of these phosphotyrosine groups serve as anchor sites for downstream signaling molecules, which themselves can also be directly phosphorylated by FGFR, leading to activation of a variety of signal transduction pathways. Thus, the MAPK signaling cascade is involved in cell growth and differentiation, the PI3K/Akt signaling cascade is involved in cell survival and cell fate decisions, and the PI3K and PKC signaling cascades function in the control of cell polarity. Several feedback inhibitors of FGF signaling have been identified to date and include members of the spry (sprouty) and Sef (expression similar to FGF) families. Furthermore, under certain conditions FGFR is released from the pre-golgi membrane into the cytosol. Receptors and their complexesThe body FGF-2 is co-transported into the nucleus by mechanisms involving import proteins and is involved in the CREB-binding protein (CBP) complex, a common and essential transcriptional co-activator that acts as a gene activation gating factor. A variety of correlations between immunohistochemical expression of FGF-2, FGFR-1 and FGFR-2 and their localization of cytoplasmic and nuclear tumor cells have been observed. For example, in lung adenocarcinoma, this association was also found at the nuclear level, emphasizing the active role of the complex in the nucleus [ Korc and Friesel,Curr. Cancer Drugs Targets5,639-651 (2009)]。

FGFs are widely expressed in developing and adult tissues and play important roles in a variety of normal and pathological processes including tissue development, tissue regeneration, angiogenesis, neoplastic transformation, cell migration, cell differentiation and cell survival. In addition, FGF as a pro-angiogenic factor has also been implicated in the emerging phenomenon of resistance to inhibition of vascular endothelial growth factor receptor-2 (VEGFR-2) [ Bergers and Hanahan,Nat. Rev. Cancer8, 592-603(2008)]。

recent cancer genomic profiles of signaling networks suggest an important role for aberrant FGF signaling in the emergence of some common human cancers [ Wesche et al,Biochem. J.437 (2), 199-213 (2011)]. Ligand-independent FGFR constitutive signaling has been described in many human cancers, such as brain, head and neck, gastric, and ovarian cancers. FGFR-mutated forms as well as FGFR-intragenic translocations have been identified in malignancies such as myeloproliferative diseases. Interestingly, the same mutations found to be responsible for many developmental disorders were also found in tumor cells (e.g., mutations found in achondroplasia and lethal dysplasia that cause dimerization of FGFR-3, leading to constitutive activation, also common in bladder cancer). Mutations that promote dimerization are only one mechanism by which ligand-independent signaling from FGFR can be increased. Other mutations located within or outside the kinase domain of FGFR can alter the conformation of the domain, resulting in a permanently active kinase.

Chromosome region 8p11-12(FGFR-1Genomic position of) ofAmplification is a common focal amplification in breast cancer and occurs in about 10% of breast cancers, mainly estrogen receptor positive cancers.FGFR-1Amplification has also been reported in non-small cell lung squamous carcinoma and is found at low incidence in ovarian cancer, bladder cancer and rhabdomyosarcoma. Similarly, about 10% of gastric cancers manifestFGFR-2Amplification, which is associated with poor prognosis, diffuse-type cancer. In addition, multiple Single Nucleotide Polymorphisms (SNPs) located in FGFR-1 to-4 were found to be associated with an increased risk of developing selective cancer, or reported to be associated with poor prognosis (e.g., FGFR-4G 388R allele in breast, colon and lung adenocarcinoma). The direct role of these SNPs in promoting cancer remains controversial.

In summary, a number of in vitro and in vivo studies have been conducted to identify FGFR-1 through-4 as important cancer targets, and a comprehensive review summarizes the results of these studies [ see, e.g., Heinzle et al,Expert Opin. Ther.Targets15(7), 829-846(2011), Wesche et al,Biochem. J.437(2), 199-,Trends in Molecular Medicine17 (5); 283-,Mol. Cancer Res.8 (11), 1439-1452 (2010)]. Several strategies have been followed to attenuate aberrant FGFR-1 to-4 signaling in human tumors, including blocking antibodies and small molecule inhibitors, among others. Several selective small molecule FGFR inhibitors are currently in clinical development, such as AZD-4547 (AstraZeneca) and BJG-398 (Novartis).

Despite significant advances in cancer therapy in general in recent years, there is a continuing need to identify new anti-cancer compounds with improved properties, such as higher potency, greater selectivity, reduced toxicity and/or better tolerability. Thus, the technical problem to be solved according to the present invention may be seen in providing alternative compounds having inhibitory activity on FGFR kinases, thereby providing new therapeutic options for the treatment of FGFR-mediated diseases, in particular cancer and other proliferative disorders.

Fused hetero-5, 6-bicyclic kinase inhibitors with 9-or 10-membered bicyclic heteroaryl substituents have been disclosed in WO2007/061737-A2 and WO 2005/097800-A1, respectively. These compounds are said to be useful in the treatment of cancer and other diseases due to their inhibitory effect on mTOR (mammalian target of rapamycin) and/or IGF-1R (insulin-like growth factor receptor type 1) kinase. Further, hetero-5, 6-bicyclic template structures associated with the inhibition of kinases have been described inter alia in WO 01/19828-A2, WO 2007/079164-A2 and WO 2010/051043-A1.

4-aminopyrrolo [2,1-f ] [1,2,4] triazine derivatives with different inhibition profiles for some protein kinases have been described inter alia in WO 00/71129-A1, WO 2007/056170-A2, WO 2007/061882-A2, WO 2007/064932-A2, WO 2009/136966-A1 and WO 2010/126960-A1.

In WO 2005/121147-A1, WO 2007/064883-A2 and WO 2007/064931-A2, 4-aminopyrrolo [2,1-f ] [1,2,4] triazine derivatives containing a substituted diaryl ureido group at the 5-position are described as having FGFR-1 inhibitory activity. However, other receptor tyrosine kinases, particularly VEGFR, PDGFR and Tie-2 kinases, are also clearly inhibited by this particular class of compounds. Since it is hypothesized that such multikinase activity may lead to an enhancement of potential side effects during treatment, it is an object of the present invention to identify new agents with improved selectivity for FGFR kinases, thereby providing new options for more tolerable cancer treatments.

Surprisingly, it has now been found that certain 4-aminopyrrolo [2,1-f ] [1,2,4] triazine derivatives carrying a specifically substituted benzothiophen-2-yl residue at the 5-position exhibit potent and selective inhibition of FGFR kinases, in particular FGFR-1 and FGFR-3 kinases, which makes these compounds particularly useful for the treatment of proliferative disorders, such as cancer and tumor diseases.

Thus, in one aspect, the present invention relates to 6, 7-disubstituted 5- (1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine derivatives of general formula (I)

Wherein

R¹Is hydrogen, chlorine, methyl or methoxy,

R²is a hydrogen or a methoxy group, and the compound is a compound represented by the formula (I),

provided that R is¹And R²At least one of which is not hydrogen,

G¹represents chlorine, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkoxycarbonyl, 5-membered azaheteroaryl or the radical-CH₂-OR³、-CH₂-NR⁴R⁵or-C (= O) -NR⁴R⁶Wherein

R³Is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₃-C₆) -cycloalkyl or phenyl, wherein

(i) Said (C)₁-C₄) Alkyl optionally substituted by hydroxy, (C)₁-C₄) -alkoxy, hydroxycarbonyl, (C)₁-C₄) Alkoxycarbonyl, amino, aminocarbonyl, mono- (C)₁-C₄) -alkylaminocarbonyl, di- (C)₁-C₄) -alkylaminocarbonyl, (C)₃-C₆) -cycloalkyl or up to three fluorine atoms,

and is

(ii) Said (C)₃-C₆) -cycloalkyl is optionally independently selected from (C)₁-C₄) -one or two substituents of alkyl, hydroxyl and amino,

and is

(iii) Said phenyl group is optionally independently selected from fluoro, chloro, bromo, cyano, trifluoromethyl, trifluoromethoxy, (C)₁-C₄) -alkyl and (C)₁-C₄) -one or two substituents of an alkoxy group,

R⁴is hydrogen or (C)₁-C₄) -an alkyl group,

R⁵is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkylcarbonyl, (C)₃-C₆) -cycloalkyl or4 to 6 membered heterocycloalkyl, wherein

(i) Said (C)₁-C₄) Alkyl optionally substituted by hydroxy, (C)₁-C₄) -alkoxy, hydroxycarbonyl, (C)₁-C₄) Alkoxycarbonyl, aminocarbonyl, mono- (C)₁-C₄) -alkylaminocarbonyl, di- (C)₁-C₄) -alkylaminocarbonyl or (C)₃-C₆) -a cycloalkyl group substitution,

and is

(iii) Said 4-to 6-membered heterocycloalkyl is optionally independently selected from (C)₁-C₄) -one or two substituents of alkyl, hydroxy, oxo and amino,

R⁶is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₃-C₆) -cycloalkyl or4 to 6 membered heterocycloalkyl, wherein

(i) Said (C)₁-C₄) Alkyl optionally substituted by hydroxy, (C)₁-C₄) -alkoxy, hydroxycarbonyl, (C)₁-C₄) Alkoxycarbonyl, amino, aminocarbonyl, mono- (C)₁-C₄) -alkylaminocarbonyl, di- (C)₁-C₄) -alkylaminocarbonyl or (C)₃-C₆) -a cycloalkyl group substitution,

and is

(ii) Said (C)₃-C₆) -cycloalkyl is optionally independently selected from (C)₁-C₄) One or two substitutions of alkyl, hydroxy and aminoThe substituent(s) is (are) substituted,

and is

or

R⁴And R⁵Or R is⁴And R⁶Are each linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 7-membered heterocycloalkyl ring which may contain a ring selected from N (R)⁷) And a second ring heteroatom of O, and may be independently selected from (C) on a ring carbon atom₁-C₄) -one or two substituents of alkyl, oxo, hydroxy, amino and aminocarbonyl, and wherein

R⁷Is hydrogen, (C)₁-C₄) -alkyl, formyl or (C)₁-C₄) -an alkyl-carbonyl group,

and is

G²Represents chlorine, cyano, (C)₁-C₄) -alkyl or a radical-CR^8AR^8B-OH、-CH₂-NR⁹R¹⁰、-C(=O)-NR¹¹R¹²or-CH₂-OR¹⁵Wherein

R^8AAnd R^8BIndependently selected from hydrogen, (C)₁-C₄) -alkyl, cyclopropyl and cyclobutyl,

R⁹is hydrogen or (C)₁-C₄) -an alkyl group,

R¹⁰is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkylcarbonyl, (C)₃-C₆) -cycloalkyl or4 to 6 membered heterocycloalkyl, wherein

(i) Said (C)₁-C₄) Alkyl optionally substituted by hydroxy, amino, aminocarbonyl, mono- (C)₁-C₄) -alkanesAminocarbonyl or di- (C)₁-C₄) -an alkyl amino carbonyl group substitution,

and is

R¹¹is hydrogen or (C)₁-C₄) -an alkyl group,

R¹²is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₃-C₆) -cycloalkyl or4 to 6 membered heterocycloalkyl, wherein

(i) Said (C)₁-C₄) Alkyl optionally substituted by hydroxy, amino, aminocarbonyl, mono- (C)₁-C₄) -alkylaminocarbonyl or di- (C)₁-C₄) -an alkyl amino carbonyl group substitution,

and is

or

R⁹And R¹⁰Or R is¹¹And R¹²Are each linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 7-membered heterocycloalkyl ringThe cycloalkyl ring may contain a group selected from N (R)¹³) O, S and S (O)₂And may be independently selected from fluorine, (C) on a ring carbon atom₁-C₄) -alkyl, oxo, hydroxy, amino and aminocarbonyl and wherein

R¹³Is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₃-C₆) -cycloalkyl, formyl or (C)₁-C₄) -an alkyl-carbonyl group,

and is

R¹⁵Is (C)₁-C₄) -an alkyl group,

with the proviso that when G²When it is chloro or cyano, G¹Is not chlorine.

The compounds of the invention may also exist in the form of their salts, solvates and/or solvates of said salts.

The compounds of the invention are compounds of the formula (I) and salts, solvates and solvates of said salts, the compounds comprised in the formulae (I-a), (I-B), (I-C), (I-D) and (I-E) of the formula (I) and salts, solvates and solvates of said salts, and the compounds comprised in the formula (I) and mentioned below as process products and/or embodiment examples and the salts, solvates and solvates of said salts, wherein the compounds comprised in the formula (I) and mentioned below are not already salts, solvates and solvates of said salts.

For the purposes of the inventionSalt (salt)Preferably a pharmaceutically acceptable salt of a compound of the invention (see, e.g., s. m. berge et al, "Pharmaceutical Salts",J. Pharm. Sci.1977, 66, 1-19). Also included are salts which are not suitable per se for pharmaceutical applications but which can be used, for example, for the isolation or purification of the compounds of the invention.

Pharmaceutically acceptable salts includeAcid addition salts of inorganic acids, carboxylic acids and sulfonic acids, for example the following: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, benzeneSulfonic acid, toluenesulfonic acid, naphthalenedisulfonic acid, formic acid, acetic acid, trifluoroacetic acid, propionic acid, lactic acid, tartaric acid, malic acid, citric acid, fumaric acid, maleic acid, and benzoic acid.

Pharmaceutically acceptable salts also includeSalts of customary bases, for example and preferably alkali metal salts (e.g. sodium and potassium salts), alkaline earth metal salts (e.g. calcium and magnesium salts) and salts derived from ammonia or organic amines (such as and preferably ethylamine, diethylamine, triethylamine, potassium salts, magnesium salts, and the like),N, NDiisopropylethylamine, monoethanolamine, diethanolamine, triethanolamine, dimethylaminoethanol, diethylaminoethanol, procaine, dicyclohexylamine, dibenzylamine,N-methylmorpholine,N-methylpiperidine, arginine, lysine and 1, 2-ethylenediamine).

In the context of the present inventionSolvatesRefers to those forms of the compounds of the present invention that form complexes in the solid or liquid state by stoichiometric coordination with solvent molecules.Hydrate of calcium and magnesiumIs a specific form of solvate in which coordination occurs with water. Preferred solvates in the context of the present invention are hydrates.

The compounds of the invention may exist (by the nature of asymmetric centers or by restricted rotation) in the form of isomers (enantiomers, diastereomers). Any isomer may exist in which the asymmetric center is (R)-、(S) -or (R,S) -configuration.

It is also understood that when two or more asymmetric centers are present in the compounds of the present invention, several diastereomers and enantiomers of the exemplified structures may generally be present, and that pure diastereomers and pure enantiomers represent preferred embodiments. Pure stereoisomers, pure diastereomers, pure enantiomers, and mixtures thereof are intended to be within the scope of the invention.

The geometric isomers arising from the nature of the substituents around the double bond or ring may be in cis (= formZ-) or trans (= g)E-) exists, and both isomeric forms are contemplatedAre within the scope of the invention.

All isomers of the compounds of the present invention, whether isolated, pure, partially pure, or racemic mixtures, are included within the scope of the present invention. Purification of the isomers and separation of the isomeric mixtures may be accomplished by standard techniques known in the art. For example, diastereomeric mixtures can be separated into the individual isomers by chromatography or crystallization, racemates can be separated into the respective enantiomers by chromatography on chiral phases or by resolution.

Furthermore, the present invention includes all possible tautomeric forms of the above compounds.

The invention also encompasses all suitable isotopic variations of the compounds of the invention. Isotopic variations of the compounds of the present invention are understood to mean compounds wherein: wherein at least one atom in a compound of the invention has been exchanged for another atom having the same atomic number, but an atomic mass different from the atomic mass usually or predominantly present in nature. Examples of isotopes that can be incorporated into the compounds of the invention are isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, chlorine, bromine and iodine, such as²H (deuterium),³H (tritium),¹³C、¹⁴C、¹⁵N、¹⁷O、¹⁸O、¹⁸F、³⁶Cl、⁸²Br、¹²³I、¹²⁴I、¹²⁹I and¹³¹I. particular isotopic variations of the compounds of the present invention, particularly those into which one or more radioactive isotopes have been incorporated, can be useful, for example, in examining the mechanism of action or the distribution of active compounds in the body. Due to the relatively easy manufacturability and detectability, use³H or¹⁴C-isotopically labelled compounds are particularly suitable for this purpose. Furthermore, the incorporation of isotopes (e.g., deuterium) can result in specific therapeutic benefits arising from greater metabolic stability of the compounds, for example increased in vivo half-life, or a reduction in the required active dose. Thus, in certain instances, such modifications of the compounds of the invention may also constituteAnd are presently preferred embodiments of the invention. Isotopic variations of the compounds of the present invention can be prepared by methods known to those skilled in the art, for example, by methods described in the procedures and working examples below, by employing specific reagents and/or corresponding isotopic modifications of the starting compounds therein.

In the context of the present invention, substituents and residues have the following meanings, unless otherwise indicated:

(C ₁ -C ₄ ) Alkyl represents in the context of the present invention a linear or branched alkyl group having from 1 to 4 carbon atoms. Can be used forMention may be made, by way of example and preferably: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

(C ₁ -C ₄ ) Alkoxy represents, in the context of the present invention, a linear or branched alkoxy group having 1 to 4 carbon atomsAnd (4) a base. Mention may be made, by way of example and preferably: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy.

Mono- (C) ₁ -C ₄ ) Alkylamino represents, in the context of the present invention, a straight chain or a cyclic alkyl group having 1 to 4 carbon atomsAmino of a branched alkyl substituent. Mention may be made, by way of example and preferably: methylamino, ethylamino, n-propylamino, isopropylamino, n-butylamino and tert-butylamino.

Di- (C) ₁ -C ₄ ) Alkylamino represents in the context of the present invention a residue having two identical or different radicals each containing 1To a linear or branched alkyl substituent of 4 carbon atoms. Mention may be made, by way of example and preferably:N,N-dimethylamino group,N,N-a diethylamino group,N-ethyl-N-methylamino radical,N-methyl-N-n-propylamino group,N-isopropyl-N-methylamino radical,N-isopropyl-N-n-propylamino group,N,N-diisopropylamino group,N-n-butyl-N-methylamino andN-tert-butyl-N-methylamino radical.

(C ₁ -C ₄ ) Alkylcarbonyl represents, in the context of the present invention, a radical prepared via carbonyl [ -C (= O) -]Bound to the remainder of the moleculeThe remainder being straight-chain or branched alkyl groups having from 1 to 4 carbon atoms. Mention may be made, by way of example and preferably: acetyl, propionyl, n-butyryl, isobutyryl, n-valeryl and pivaloyl.

(C ₁ -C ₄ ) Alkoxycarbonyl represents, in the context of the present invention, a carbonyl [ -C (= O) -]Bound to moleculesThe remaining part of linear or branched alkoxy groups having 1 to 4 carbon atoms. Mention may be made, by way of example and preferably: methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, isopropoxycarbonyl, n-butoxycarbonyl and tert-butoxycarbonyl.

Mono- (C) ₁ -C ₄ ) Alkylaminocarbonyl represents, in the context of the present invention, a compound which is prepared via carbonyl [ -C (= O) -]Is bonded toThe remainder of the molecule and having an amino group with a straight or branched alkyl substituent containing 1 to 4 carbon atoms. Mention may be made, by way of example and preferably: methylaminocarbonyl, ethylaminocarbonyl, n-propylaminocarbonyl, isopropylaminocarbonyl, n-butylaminocarbonyl and tert-butylaminocarbonyl.

Di- (C) ₁ -C ₄ ) Alkylaminocarbonyl represents, in the context of the present invention, a compound which is prepared via carbonyl [ -C (= O) -]Is bonded toThe remainder of the subgroups and amino groups having two identical or different straight-chain or branched alkyl substituents having in each case 1 to 4 carbon atoms. Mention may be made, by way of example and preferably:N,N-dimethylaminocarbonyl group,N,N-diethylaminocarbonyl,N-ethyl radical-N-methylaminocarbonyl group,N-methyl-N-n-propylaminocarbonyl group,N-isopropyl-N-methylaminocarbonyl group,N, N-diisopropylaminocarbonyl group,N-n-butyl-N-methylaminocarbonyl andN-tert-butyl-N-methylaminocarbonyl.

(C ₃ -C ₆ ) Cycloalkyl in the context of the present invention represents a monocyclic saturated carbocyclic ring having 3 to 6 ring carbon atoms.Mention may be made, by way of example, of: cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Preferred are cyclopropyl and cyclobutyl.

4 to 7 membered heterocycloalkylAnd4-to 6-membered heterocycloalkylIn the context of the present invention, denotes monocyclic saturated heterocycles having 4 to 7 or4 to 6 total ring atoms, respectively, which contain one or two identical or different rings from the series N, O, S and S (O)₂And may be bonded via a ring carbon atom or via a ring nitrogen atom, if present. Containing a ring nitrogen atom and optionally from the series N, O or S (O)₂A 4 to 6 membered heterocycloalkyl group of one other ring heteroatom is preferred. Containing one ring nitrogen atom and optionally one other ring hetero atom from the series N or O5-or 6-membered heterocycloalkylIs particularly preferred. Mention may be made, by way of example, of: azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrofuranyl, thietanyl (thiolanyl), 1-thiabendanyl, 1, 2-oxazolidinyl, 1, 3-thiazolidinyl, piperidinyl, piperazinyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1, 3-dioxanyl, 1, 4-dioxanyl, 1, 2-oxazinyl (1,2-oxazinanyl), morpholinyl, thiomorpholinyl, 1-thiomorpholinyl, azepanyl, 1, 4-diazepanyl, and 1, 4-oxazepanyl. Preference is given to azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, 1, 2-oxazolidinyl, 1, 3-oxazolidinyl, piperidinyl, piperazinyl, 1, 2-oxazinidinyl, morpholinyl and thiomorpholinyl. Particularly preferred are pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl.

5-membered aza-heteroarylIn the context of the present invention, represents an aromatic heterocyclic group (heteroaromatic) having 5 total ring atoms, which contains at least one ring nitrogen atom and optionally one or two further ring heteroatoms from the series N, O and/or S and is bonded via a ring carbon atom or optionally via a ring nitrogen atom (when valency permits). 5-membered aza-heteroaryl groups containing one ring nitrogen atom and one or two other ring heteroatoms from the series N and/or O are preferred. Mention may be made, by way of example, of: pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, triazolyl, oxadiazolyl and thiadiazolyl. Preferred are pyrazolyl, imidazolyl, oxazolyl, isoxazolyl and oxadiazolyl.

Oxo-substituted radicalIn the context of the present invention represents an oxygen atom bonded to a carbon atom via a double bond.

In the context of the present invention, the meaning is independent of one another for all multiply occurring radicals. If a group in a compound of the invention is substituted, the group may be mono-or polysubstituted, unless otherwise specified. Preference is given to substitution by one or by two or three identical or different substituents. Substitution by one or by two identical or different substituents is particularly preferred.

In a preferred embodiment, the invention relates to compounds of the general formula (I), in which

R¹Is a chlorine, a methyl or a methoxy group,

G¹represents chlorine, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -an alkoxycarbonyl group or a 5-membered azaheteroaryl group selected from pyrazolyl, imidazolyl, oxazolyl, isoxazolyl and oxadiazolyl, or represents the group-CH₂-OR³or-CH₂-NR⁴R⁵Wherein

R³Is hydrogen, (C)₁-C₄) -alkyl or (C)₃-C₆) -a cycloalkyl group,

wherein said (C)₁-C₄) Alkyl optionally substituted by hydroxy, (C)₁-C₄) -alkoxy, hydroxycarbonyl, (C)₁-C₄) Alkoxycarbonyl, amino, aminocarbonyl, (C)₃-C₆) -cycloalkyl or up to three fluorine atoms,

R⁴is hydrogen or (C)₁-C₄) -an alkyl group,

R⁵is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkylcarbonyl, (C)₃-C₆) -cycloalkyl or 5-or 6-membered heterocycloalkyl, wherein

(i) Said (C)₁-C₄) Alkyl optionally substituted by hydroxy, hydroxycarbonyl or (C)₃-C₆) -a cycloalkyl group substitution,

and is

(ii) Said 5-or 6-membered heterocycloalkyl being optionally substituted by oxo,

or

R⁴And R⁵Are linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 6-membered heterocycloalkyl ring which may contain a ring selected from N (R)⁷) And O, and may be substituted on a ring carbon atom by oxo or hydroxy, and wherein

R⁷Is hydrogen or (C)₁-C₄) -an alkyl group,

and is

R^8AAnd R^8BIndependently of each otherSelected from hydrogen, (C)₁-C₄) -an alkyl group and a cyclopropyl group,

R⁹is hydrogen or a methyl group, or a mixture thereof,

R¹⁰is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkylcarbonyl, (C)₃-C₆) -cycloalkyl or 5-or 6-membered heterocycloalkyl, wherein

(i) Said (C)₁-C₄) -alkyl is optionally substituted by hydroxy or aminocarbonyl,

and is

(ii) Said 5-or 6-membered heterocycloalkyl being optionally substituted by oxo,

R¹¹is hydrogen or a methyl group, or a mixture thereof,

R¹²is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₃-C₆) -cycloalkyl or 5-or 6-membered heterocycloalkyl, wherein

(i) Said (C)₁-C₄) -the alkyl group is optionally substituted by a hydroxyl group,

and is

(ii) Said 5-or 6-membered heterocycloalkyl being optionally substituted by oxo,

or

R⁹And R¹⁰Or R is¹¹And R¹²Are each linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 6-membered heterocycloalkyl ring which may contain a ring selected from N (R)¹³) O, S and S (O)₂And may be independently selected from fluorine, (C) on a ring carbon atom₁-C₄) -alkyl, oxo, hydroxy, amino and aminocarbonyl and wherein

R¹³Is hydrogen, (C)₁-C₄) Alkyl, cyclopropyl, cyclobutyl, formyl or (C)₁-C₄) -an alkyl-carbonyl group,

and is

R¹⁵Is a methyl group or an ethyl group,

with the proviso that when G²When it is chloro or cyano, G¹Is not chlorine.

In a particularly preferred embodiment, the invention relates to compounds of the general formula (I), in which

R¹Is a methyl group, and the compound is,

R²is a methoxy group, and the compound is a methoxy group,

G¹represents methyl, oxazol-5-yl or a radical-CH₂-OR³or-CH₂-NR⁴R⁵Wherein

R³Is hydrogen, (C)₁-C₄) -an alkyl, cyclopropyl or cyclobutyl group,

wherein said (C)₁-C₄) -alkyl is optionally substituted by hydroxy, methoxy, ethoxy, hydroxycarbonyl, methoxycarbonyl, ethoxycarbonyl, amino, aminocarbonyl, cyclopropyl, cyclobutyl or up to three fluorine atoms,

R⁴is hydrogen, a methyl group or an ethyl group,

R⁵is hydrogen, (C)₁-C₄) -alkyl, acetyl, cyclopropyl, cyclobutyl or 2-oxopyrrolidin-3-yl,

wherein said (C)₁-C₄) -alkyl is optionally substituted by hydroxy, hydroxycarbonyl, cyclopropyl or cyclobutyl,

or

R⁴And R⁵Are linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 5-or 6-membered heterocycloalkyl ring which may contain a second ring heteroatom selected from NH and O and which may be substituted on a ring carbon atom by oxo or hydroxy,

and is

G²Represents methyl or a radical-CR^8AR^8B-OH、-CH₂-NR⁹R¹⁰or-C (= O) -NR¹¹R¹²Wherein

R^8AAnd R^8BIndependently of one another is hydrogen or a methyl group,

R⁹is a hydrogen atom, and is,

R¹⁰is hydrogen, (C)₁-C₄) -alkyl, acetyl, cyclopropyl, cyclobutyl or 2-oxopyrrolidin-3-yl,

wherein said (C)₁-C₄) -alkyl is optionally substituted by hydroxy or aminocarbonyl,

R¹¹is hydrogen or a methyl group, or a mixture thereof,

R¹²is hydrogen, (C)₁-C₄) -alkyl, cyclopropyl, cyclobutyl or 2-oxopyrrolidin-3-yl,

wherein said (C)₁-C₄) -the alkyl group is optionally substituted by a hydroxyl group,

or

R⁹And R¹⁰Or R is¹¹And R¹²Are each linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 6-membered heterocycloalkyl ring which may contain a ring selected from N (R)¹³) O and S (O)₂And may be substituted on a ring carbon atom with up to three substituents independently selected from fluorine, methyl, oxo, hydroxy, amino and aminocarbonyl, and wherein

R¹³Is hydrogen, formyl or acetyl.

In a particular embodiment, the invention relates to compounds of the general formula (I), wherein

R¹Is a methyl group, and the compound is,

and is

R²Is methoxy.

In a further unique embodiment, the present invention relates to a compound of formula (I), wherein

G¹Represents the group-CH₂-OR³Wherein

R³Is hydrogen or (C)₁-C₄) -alkyl optionally substituted with hydroxy, methoxy, amino, aminocarbonyl or up to three fluorine atoms.

In another unique embodiment, the present invention relates to compounds of formula (I), wherein

G¹Represents the group-CH₂-NR⁴R⁵Wherein

R⁴Is hydrogen or a methyl group, or a mixture thereof,

R⁵is (C)₁-C₄) -alkyl, acetyl, cyclopropyl, cyclobutyl or 2-oxopyrrolidin-3-yl,

or

R⁴And R⁵Are linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 5-or 6-membered heterocycloalkyl ring which may contain a second ring heteroatom selected from NH and O and which may be substituted on a ring carbon atom by oxo or hydroxy.

G²Represents the group-CH₂-NR⁹R¹⁰Wherein

R⁹Is a hydrogen atom, and is,

R¹⁰is acetyl or 2-oxopyrrolidin-3-yl,

or

R⁹And R¹⁰Are linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 5-or 6-membered heterocycloalkyl ring which may contain a ring selected from N (R)¹³) And O, and may be substituted on a ring carbon atom with up to two substituents independently selected from methyl, oxo, hydroxy and amino, and wherein

R¹³Is hydrogen, formyl or acetyl.

In yet another unique embodiment, the present invention relates to a compound of formula (I), wherein

G²Represents a group-C (= O) -NR¹¹R¹²Wherein

R¹¹Is a hydrogen atom, and is,

R¹²is (C)₁-C₄) -alkyl or 2-oxopyrrolidin-3-yl,

or

R¹¹And R¹²Are linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 6-membered heterocycloalkyl ring which may contain a second ring heteroatom selected from NH and O and which may be substituted on a ring carbon atom by oxo or hydroxy.

R¹Is a methyl group, and the compound is,

R²is a methoxy group, and the compound is a methoxy group,

G¹represents the group-CH₂-OR³Wherein

R³Is optionally substituted by hydroxy, amino or aminocarbonyl₁-C₄) -an alkyl group,

and is

G²Represents the group-CH₂-NR⁹R¹⁰or-C (= O) -NR¹¹R¹²Wherein

R⁹Is a hydrogen atom, and is,

R¹⁰is a 2-oxopyrrolidin-3-yl group,

or

R⁹And R¹⁰Are linked and together with the nitrogen atom to which they are attached form a piperazin-1-yl, 3-oxopiperazin-1-yl or 4-acetylpiperazin-1-yl ring,

R¹¹is a hydrogen atom, and is,

R¹²is a 2-oxopyrrolidin-3-yl group,

or

R¹¹And R¹²Are linked and together with the nitrogen atom to which they are attached form a 3-hydroxyazetidin-1-yl, 4-hydroxypiperidin-1-yl or 3-oxopiperazin-1-yl ring.

The definitions of residues specifically identified in each combination or preferred combination of residues may also be replaced, as desired, with definitions of residues from other combinations, regardless of the particular combination identified for the residues. Combinations of two or more of the above preferred ranges are particularly preferred.

The compounds of formula (I) may be represented by a particular G group which is selected primarily¹And G²The nature of the group (see definitions above) dictates various synthetic routes.

Thus, in another embodiment, the invention relates to a process for the preparation of compounds of the general formula (I), characterized in that

[A] 6-substituted 4-aminopyrrolo [2,1-f ] [1,2,4] triazines of formula (II)

Wherein R is³Has the meaning of the above-mentioned formula,

first with formaldehyde and an amine of the formula (III) in the presence of an acid

Wherein R is⁹And R¹⁰Has the meaning of the above-mentioned formula,

to give a compound of the formula (IV)

Wherein R is³、R⁹And R¹⁰Has the meaning of the above-mentioned formula,

then brominated to the compound of formula (V)

Wherein R is³、R⁹And R¹⁰Has the meaning of the above-mentioned formula,

and subsequently coupled with a boronic acid benzothiophen-2-yl ester of formula (VI) in the presence of a palladium catalyst and a base

Wherein R is¹And R²Has the meaning of the above-mentioned formula,

and is

R¹⁴Represents hydrogen or (C)₁-C₄) -alkyl, or twoR¹⁴The residues are linked together to form- (CH)₂)₂-、-C(CH₃)₂-C(CH₃)₂-、-(CH₂)₃-、-CH₂-C(CH₃)₂-CH₂-or-C (= O) -CH₂-N(CH₃)-CH₂-a C (= O) -bridge,

to obtain the target compound of formula (I-A)

Wherein R is¹、R²、R³、R⁹And R¹⁰Has the meaning of the above-mentioned formula,

or

[B] 6-substituted 4-aminopyrrolo [2,1-f ] [1,2,4] triazines of formula (II)

Wherein R is³Has the meaning of the above-mentioned formula,

first in the presence of phosphorus oxychlorideN,NFormylation of dimethylformamide to give aldehydes of the formula (VII)

Wherein R is³Has the meaning of the above-mentioned formula,

then brominated to the compound of formula (VIII)

Wherein R is³Has the meaning of the above-mentioned formula,

Wherein R is¹、R²And R¹⁴Has the meaning of the above-mentioned formula,

to give a compound of formula (IX)

Wherein R is¹、R²And R³Has the meaning of the above-mentioned formula,

then put it in

[ B-1] reaction with an amine of the formula (III) in the presence of an acid and a reducing agent

Wherein R is⁹And R¹⁰Has the meaning of the above-mentioned formula,

to obtain the target compound of formula (I-A)

or

[ B-2] Oxidation to carboxylic acids of formula (X)

Wherein R is¹、R²And R³Has the meaning of the above-mentioned formula,

and finally with amines of the formula (XI) in the presence of condensing agents

Wherein R is¹¹And R¹²Has the meaning of the above-mentioned formula,

to obtain the target compound of formula (I-B)

Wherein R is¹、R²、R³、R¹¹And R¹²Has the meaning of the above-mentioned formula,

or

[C] 6-substituted 4-amino-5-bromopyrrolo [2,1-f ] [1,2,4] triazines of formula (XII)

First coupled with a boronic acid benzothiophen-2-yl ester of formula (VI) in the presence of a palladium catalyst and a base

Wherein R is¹、R²And R¹⁴Has the meaning of the above-mentioned formula,

to give a compound of the formula (XIII)

Wherein R is¹And R²Has the meaning of the above-mentioned formula,

then reacted with formaldehyde and an amine of formula (III) in the presence of an acid

Wherein R is⁹And R¹⁰Has the meaning of the above-mentioned formula,

to give a compound of the formula (I-C)

Wherein R is¹、R²、R⁹And R¹⁰Has the meaning of the above-mentioned formula,

then it is put in

[ C-1] Oxidation to aldehydes of formula (XIV)

treatment with an amine of formula (XV) in the presence of an acid and a reducing agent

Wherein R is⁴And R⁵Has the meaning of the above-mentioned formula,

to obtain the target compound of formula (I-D)

Wherein R is¹、R²、R⁴、R⁵、R⁹And R¹⁰Has the meaning of the above-mentioned formula,

or

[ C-2] to the corresponding 6- (halomethyl) derivative of formula (XVI)

and is

X is chlorine, bromine or iodine,

and treatment with an alcohol of formula (XVII) in the presence of a base

Wherein R is^3AHaving the above-mentioned R³The meaning of (other than hydrogen),

to obtain the target compound of formula (I-E)

Wherein R is¹、R²、R^3A、R⁹And R¹⁰Has the meaning of the above-mentioned formula,

then optionally, if appropriate (i) The compounds of formula (I) thus obtained are preferably separated into their respective enantiomers and/or diastereomers by chromatographic methods, and/orii) The compounds of formula (I) are converted into their respective hydrates, solvates, salts and/or hydrates or solvates of the salts by treatment with the corresponding solvents and/or acids or bases.

The compounds of formula (I-A), (I-B), (I-C), (I-D) and (I-E) which can be prepared by the above-described process each represent a particular subset of the compounds of formula (I).

Process steps [ a ] (II) → (IV) and [ C ] (XIII) → (I-C), representing a mannich type aminomethylation reaction, are carried out in the usual manner by treating the respective starting compounds with a mixture of aqueous formaldehyde and an amine component (III) in the presence of an acid catalyst such as formic acid or acetic acid. Preferably, acetic acid is used as both catalyst and solvent. The reaction is generally carried out at a temperature in the range of +20 ℃ to +80 ℃.

As for method step [ A](IV) → (V) and [ B-](VII) → (VIII) brominating agent, preferably usedN-bromosuccinimide (NBS), 1, 3-dibromo-5, 5-dimethylhydantoin (DBDMH) or elemental bromine. The reaction is usually carried out at a temperature in the range of-78 ℃ to +20 ℃ in an inert solvent such as dichloromethane, chloroform, tetrahydrofuran, acetonitrile orN,N-Dimethylformamide (DMF).

Coupling reaction [ A ]](V) + (VI) → (I-A)、[B](VIII) + (VI) → (IX) and [ C](XII) + (VI) → (XIII) [ "Suzuki-Miyaura coupling"]Usually in an inert solvent with the aid of a palladium catalyst and an aqueous base. Palladium catalysts suitable for this purpose include, for example, palladium (II) acetate, palladium (II) chloride, bis (triphenylphosphine) palladium (II) chloride, bis (acetonitrile) palladium (II) chloride, [1,1' -bis (diphenylphosphino) ferrocene]Palladium (II) chloride, tetrakis (triphenylphosphine) palladium (0), bis (dibenzylideneacetone) palladium (0) and tris (dibenzylideneacetone) dipalladium (0), optionally in combination with other phosphine ligands, such as 2-dicyclohexylphosphino-2 ',4',6 '-triisopropylbiphenyl (X-Phos), 2-dicyclohexylphosphino-2'6' -dimethoxybiphenyl (S-Phos), 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (xanthphos) or 4- (di-tert-butylphosphino) -N,N-dimethylaniline. In addition, palladium precatalysts from which catalytically active species are formed under the reaction conditions may be used, such as ((2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine [ see, for example, s. Kotha et al,Tetrahedron58, 9633, 9695 (2002), T.E. Barder et al,J.Am.Chem. Soc.127 (13), 4685-4696 (2005); S.L. Buchwald et al, J. Am. Chem.Soc.132 (40), 14073-]。

Suitable bases for these coupling reactions are in particular alkali metal carbonates, such as sodium carbonate, potassium carbonate or cesium carbonate, alkali metal phosphates, such as sodium phosphate or potassium phosphate, or alkali metal fluorides, such as potassium fluoride or cesium fluoride. Typically, these bases are used in the form of aqueous solutions. The reaction is carried out in an organic solvent which is inert under the reaction conditions. Preferably, a water-miscible organic solvent is used, such as 1, 2-dimethoxyethane, tetrahydrofuran, 1, 4-dioxane, acetonitrile,N,NDimethylformamide (DMF) or Dimethylsulfoxide (DMSO), but other inert solvents such as dichloromethane or toluene may also be used.

Method step [ B](II) → (VII) [ "Vilsmeier-Haack formylation"]By treatment with phosphorus oxychlorideN, NPyrrolotriazine (II) in Dimethylformamide (DMF) solvent is carried out in the usual manner. The reaction is generally carried out at a temperature of from 0 ℃ to +80 ℃.

Suitable reducing agents for the reductive amination reactions [ B-1] (IX) + (III) → (I-A) and [ C-1] (XIV) + (XV) → (I-D) are the customary alkali metal borohydrides, such as lithium borohydride, sodium borohydride, potassium borohydride, sodium cyanoborohydride or sodium triacetoxyborohydride. Depending on the reactivity of each of the amine components (iii) and (XV) and/or the particular borohydride used, the conversion is generally carried out in an alcohol or ether solvent, such as methanol, ethanol, isopropanol, tetrahydrofuran or 1, 4-dioxane, in the presence of an acid, preferably acetic acid, at a temperature in the range of from 0 ℃ to +80 ℃.

For method step [ B-2]Oxidation In (IX) → (X), oxidation with sodium chlorite in the presence of a hypochlorite scavenger such as 2-methyl-2-butene represents the method of choice [ see h.w. Pinnick et al,Tetrahedron37, 2091-,J. Prakt. Chem.342(6), 605-]. The reaction is generally carried out at a temperature of from 0 ℃ to room temperature in a tetrahydrofuran/water mixture.

Is suitable for use in method step [ B-2](X) + (XI) → (I-B) [ amide formation]The condensing agent of (A) includes, for example, a carbodiimide such asN,N'-a diethyl group-,N,N'-a dipropyl group-,N,N'-a diisopropyl group-,N,N'dicyclohexylcarbodiimide (DCC) orN- (3-dimethylaminopropyl) -N'-Ethylcarbodiimide (EDC), phosgene derivatives, such asN,N'Carbonyldiimidazole (CDI) or isobutyl chloroformate, α -chloroalkenamine, such as 1-chloro-2-methyl-1-dimethylamino-1-propene, phosphorus compounds, such as propanephosphonic anhydride, diethyl cyanophosphonate, bis (2-oxo-3-oxazolidinyl) phosphoryl chloride, benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate (BOP) or benzotriazol-1-yloxy-tris (pyrrolidino) phosphonium hexafluorophosphate (PyBOP), and urea cation compounds, such asO- (benzotriazol-1-yl) -N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU),O- (benzotriazol-1-yl) -N,N,N',N'Tetramethyluronium Hexafluorophosphate (HBTU), 2- (2-oxo-1- (2)H) -pyridyl) -1,1,3, 3-tetramethyluronium tetrafluoroborate (TPTU),O- (7-azabenzotriazol-1-yl) -N,N, N',N'Tetramethylurea Hexafluorophosphate (HATU) orO-(1H6-chlorobenzotriazol-1-yl) -1,1,3, 3-tetramethyluronium tetrafluoroborate (TCTU), if appropriate in combination with further auxiliaries, such as 1-hydroxybenzotriazole (HOBt) orN-hydroxysuccinimide (HOSu), and/or a base such as an alkali metal carbonate, for example sodium or potassium carbonate, or an organic amine base such as triethylamine,N-a methyl-piperidine compound having a structure represented by formula (I),N-methylmorpholine (NMM),N,Ndiisopropylethylamine (DIPEA), pyridine or 4-N,N-Dimethylaminopyridine (DMAP). Is preferably usedO- (7-azabenzotriazol-1-yl) -N,N,N',N'Tetramethylurea Hexafluorophosphate (HATU) orO- (benzotriazol-1-yl) -N,N,N',N'Combinations of tetramethyluronium tetrafluoroborates (TBTU)N,N-Diisopropylethylamine (DIPEA) and optionally 1-hydroxybenzotriazole (HOBt).

For method step [ B-2]Inert solvents of (X) + (XI) → (I-B) are, for example, ethers such as diethyl ether, tert-butyl methyl ether, tetrahydrofuran, 1, 4-dioxane or 1, 2-dimethoxyethane, hydrocarbons such as benzene, toluene, xylene, hexane or cyclohexane, halogenated hydrocarbons such as dichloromethane, trichloromethane, carbon tetrachloride, 1, 2-dichloroethane, trichloroethylene or chlorobenzene, or other solvents such as acetone, acetonitrile, ethyl acetate, pyridine, dimethyl sulfoxide (DMSO)N,N-Dimethylformamide (DMF),N,N'-Dimethylpropyleneurea (DMPU) orN-methylpyrrolidone (NMP). Mixtures of these solvents may also be used. Preference is given to using methylene chloride, tetrahydrofuran,N,N-dimethylformamide or mixtures thereof. The reaction is generally carried out at a temperature in the range of from 0 ℃ to +60 ℃, preferably from +10 ℃ to +40 ℃.

Capable of converting primary alcohols (I-C) to aldehydes (XIV) under mild conditions (Process [ C-1]]) The oxidant includes 1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3 (1)H) -ketones ("Dess-Martin periodinane"), 2,6, 6-tetramethylpiperidine-1-oxide (TEMPO) in combination with a second oxidizing agent such as iodosobenzene-I,IDiacetate salts or sodium hypochlorite, and an oxidation system based on dimethyl sulfoxide (DMSO), such as DMSO/trifluoroacetic anhydride or DMSO-N,N'Dicyclohexylcarbodiimide (DCC). Preferred is 1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1 ziodoxol)H) -a ketone. The reaction is generally carried out in an inert solvent, preferably using dichloromethane.

For the hydroxyl-to-halogen conversion in process step [ C-2] (I-C) → (XVI), various standard methods and reagents well known in the art can be used. The reagents selected were thionyl chloride [ for X = Cl ], tetrabromomethane/triphenylphosphine [ for X = Br ], and iodine/triphenylphosphine [ for X = I ]. For reasons of ease of work-up and stability of the compounds, the preparation of the 6- (chloromethyl) derivative (XVI) [ X = Cl ] is preferred.

Is suitable for use in process step [ C-2](XVI) + (XVII) → (I-E) [ Ether formation-]Are in particular alkali metal carbonates, such as lithium carbonate, sodium carbonate, potassium carbonate or cesium carbonate, alkali metal acetates, such as sodium acetate or potassium acetate, or conventional tertiary amine bases, such as triethylamine,N-methylmorpholine,N-methylpiperidine, N-acetylsalicylic acid,N,N-diisopropylethylamine or pyridine. It is preferable thatN,NDiisopropylethylamine (DIPEA). The reaction (XVI) + (XVII) → (I-E) is carried out in a temperature range of +20 ℃ to +200 ℃, preferably +50 ℃ to +150 ℃ in an inert solvent such as tetrahydrofuran or without solvent using an excess of the alcohol (XVII). Advantageously, the conversion is carried out by means of a microwave reactor device.

The reaction sequence (I-C) → (XVI) → (I-E) may be carried out in two separate steps, i.e. isolation and purification of the intermediate compound (XVI), or it may be carried out using a one-pot procedure, i.e. using the crude intermediate (XVI) as obtained in the preparation reaction.

If the primary or secondary amine moiety forms G in the target compound of the formula (I)¹Or G²It is sometimes appropriate to use a protected derivative of such an amine as part of the group in the above preparation reaction, without using the free amine as a reaction component. For this purpose, conventional temporary amino protecting groups can be employed, such as acyl groups (e.g., acetyl or trifluoroacetyl) or carbamate-type protecting groups (e.g., Boc-, Cbz-or Fmoc-groups). Preferably, a Boc (tert-butoxycarbonyl) group is used. Similarly, as G¹Or G²The hydroxyl function of a part of the group can be temporarily blocked in precursor compounds and process intermediates, for example as Tetrahydropyranyl (THP) ether or as a silyl ether derivative, such as trimethylsilyl ether or tert-butyldimethylsilyl ether.

These protecting groups may then be present inThe aqueous workup and purification procedures are cleaved off simultaneously during the course of their removal or they may be removed in a subsequent separate reaction step using standard methods well known in the art. Likewise, the preparation of such protected intermediates of the corresponding free amines or alcohols can be readily achieved following the general procedures described in the literature [ see, e.g., t.w. Greene and p.wuts,Protective Groups in Organic Synthesis, Wiley, NewYork, 1999]。

certain types of protected (i.e., acylated) amine derivatives exert significant FGFR inhibitory activity by themselves. Accordingly, the general formula (I) as defined above also includes such compounds.

The preparation of the compounds of the invention can be illustrated by the following reaction scheme:

。

the 6-substituted 4-aminopyrrolo [2,1-f ] of formula (II) can be prepared, for example, by two different routes as described in scheme 5 below][1,2,4]A triazine. In the first pathway, 4-amino-6-cyanopyrrolo [2,1-f ] is produced by acid-mediated alcoholysis][1,2,4]Triazine (XVIII) is converted to ester (XIX) and then reduced to 6- (hydroxymethyl) compound (IIa) [ (II) R using lithium triethylborohydride³= H]. Standard conversion to the corresponding 6- (halomethyl) pyrrolotriazines, such as the chlorine compound (XX), followed by reaction with a base in the presence of a baseThe alcohol treatment of the formula (XVII) readily provides the ether derivative of the formula (IIb) [ (II) R³≠ H]. The preparation of the starting compound (XVIII) has been previously described [ see international patent application WO2007/064883 (intermediate AX/step 3).

The second route starts from the protected 1-amino-4-bromo-2-cyanopyrrole (XXI) [ preparation given in International patent application WO2007/064883 (intermediate AAE, step 3)]. Deprotonation of the urethane nitrogen, followed by metallation at the 4-position and reaction with formaldehyde gives the 4- (hydroxymethyl) derivative (XXII). Treatment with hydrogen chloride followed by addition of alcohol (XVII) and condensation with formamidine then provides the target compound of formula (IIb) using a one-pot procedure. This route is particularly suitable for the preparation of R in the alkyl ether derivatives [ (IIb)^3A= (C₁-C₄) -alkyl radical]Since alcohol reactant (XVII) can also act as a reaction solvent in these cases.

4-amino-5-bromopyrrolo [2,1-f ] [1,2,4] triazine derivatives of formula (XII) can be readily obtained from 4-amino-6- (hydroxymethyl) pyrrolo [2,1-f ] [1,2,4] triazine (IIa) by initial 5, 7-dibromination with 1, 3-dibromo-5, 5-dimethylhydantoin and subsequent selective 7-debromination via halogen-metal exchange with n-butyllithium and subsequent quenching with methanol (see scheme 6 below) (see scheme 5).

The benzothien-2-yl boronic acid esters of formula (VI) can be conveniently prepared starting from substituted thiophenol derivatives of formula (XXIV) (see scheme 7 below). Alkylation with bromoacetaldehyde diethyl acetal (XXV) and subsequent polyphosphoric acid-mediated cyclization provides a benzothiophene intermediate of formula (XXVII) which is then metallated at the 2-position and reacted with a trialkyl borate. After alkali treatmentThe treatment gives free (benzothien-2-yl) boronic acids of the formula (VIa), which can, if desired, be converted into cyclic boronic esters by standard procedures known in the art, for example the so-called MIDA boronic esters of the formula (VIb) [ see, for example, d.m. Knapp et al,J. Am. Chem. Soc.131 (20), 6961-6963 (2009)]。

[cfp.a. ple and l.j. Marnett,J. Heterocyclic Chem.25 (4), 1271-,J. Med. Chem.50 (23), 5644-5654 (2007)]。

the compounds of formulae (III), (XI), (XV), (XVII), (XXIV) and (XXV) are either commercially available, known from the literature, or can be readily prepared from readily available starting materials by employing standard procedures described in the literature. Detailed procedures and references for the preparation of starting materials can also be found in the experimental part of the starting materials and intermediate preparation section.

A further subset of the preparation of compounds of formula (I) is described in reaction schemes 8-14 below. The desired pyrrolotriazine precursors can be readily synthesized by conventional methods well known in the art, and further synthetic transformations, in most cases following the preparative routes already outlined in the methods section above, using similar reaction types, e.g., bromination, boronic ester coupling, aminomethylation, reductive amination, oxidation, and/or ether or amide formation reactions. Further details regarding the exemplary embodiments and the preparation of their respective precursor compounds are provided in the experimental section.

The compounds of the present invention possess valuable pharmacological properties and are useful in the prevention and treatment of conditions in humans and other mammals.

The compounds of the invention are potent inhibitors of the activity or expression of receptor tyrosine kinases, particularly FGFR kinases, and most notably FGFR-1 and FGFR-3 kinases. Thus, in another embodiment, the invention provides a method of treating a condition involving or mediated by FGFR kinase activity in a patient in need of such treatment, which comprises administering to said patient an effective amount of a compound of formula (I) as defined above. In certain embodiments, the disorder in which FGFR kinase activity is implicated is a proliferative disorder, particularly cancer and tumor diseases.

In the context of the present invention, the term "treating" or "treatment" includes inhibiting, delaying, slowing, alleviating, arresting, reducing, or resolving a disease, disorder, condition, or condition, its occurrence and/or progression, and/or its symptoms. The term "prevention" or "preventing" includes reducing the risk of having, infecting, or experiencing a disease, disorder, condition, or illness, the occurrence and/or progression thereof, and/or the symptoms thereof. The term prevention (prevention) includes prevention (prophyxiases). The treatment or prevention of a disorder, disease, condition, or disease state may be partial or complete.

The term "proliferative disorder" includes disorders involving undesired or uncontrolled proliferation of cells. The compounds of the present invention may be used to prevent, inhibit, block, reduce, control, etc., cell proliferation and/or cell division, and/or produce apoptosis. Such methods comprise administering to an individual in need thereof (including mammals, including humans) an amount of a compound of the present invention, or a pharmaceutically acceptable salt, isomer, polymorph, metabolite, hydrate or solvate thereof, effective to treat or prevent the disorder.

Throughout this document, it is preferred to use the singular language rather than the plural language for brevity, but singular language is generally intended to include the plural language if not otherwise stated. For example, the expression "a method of treating a disease in a patient, said method comprising administering to the patient an effective amount of a compound of formula (I)" is intended to include treating more than one disease simultaneously and administering more than one compound of formula (I).

Proliferative disorders that can be treated and/or prevented with the compounds of the present invention include, but are not limited to, inter alia, the group of cancer and tumor diseases. These are understood to mean, in particular, the following diseases, but not limited to them:

breast and mammary tumors (ductal and lobular forms, also in situ), tumors of the respiratory tract (small-cell and non-small-cell lung carcinoma, small-cell and non-small-cell carcinoma, bronchial adenoma, pleuropulmonary blastoma), tumors of the brain (e.g., brain tumors of the brain stem and hypothalamus, astrocytomas, glioblastomas, ependymomas, and neuroectodermal and pineal tumors), tumors of the digestive organs (esophagus, stomach, gall bladder, small intestine, large intestine, rectum, anus), tumors of the liver (in particular hepatocellular carcinoma, cholangiocellular carcinoma and mixed hepatocyte and cholangiocellular carcinoma), tumors of the head and neck (larynx, hypopharynx, nasopharynx, oropharynx, lips, and oral cavity), tumors of the skin (squamous epithelial carcinoma, kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer, and non-melanoma skin cancers), tumors of the soft tissue (in particular sarcomas, osteosarcoma, malignant fibrous histiocytoma, lymphosarcoma and rhabdomyosarcoma), eye tumors (in particular intraocular melanoma, uveal melanoma and retinoblastoma), tumors of endocrine and exocrine glands (such as thyroid and parathyroid, pancreas and salivary glands), tumors of urinary tract (tumors of bladder, penis, kidney, renal pelvis and ureter), tumors of reproductive organs (carcinomas of the endometrium, cervix, ovary, vagina, vulva and uterus of women, and prostate and testis of men), and distant metastases thereof. These conditions also include proliferative blood diseases in solid form and as circulating blood cells, such as lymphomas, leukemias and myeloproliferative diseases, e.g., acute myelogenous, acute lymphoblastic, chronic lymphocytic, chronic myelogenous and hairy cell leukemia and AIDS-related lymphomas, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, Burkitt's lymphoma, and lymphomas in the central nervous system.

Due to their activity and selectivity profile, the compounds of the invention are believed to be particularly suitable for the treatment of cancer and neoplastic diseases of the breast (breast), lung, stomach (stomach), bladder and ovary. Furthermore, the compounds of the invention may in general be particularly suitable for preventing or inhibiting tumor metastasis.

Other proliferative disorders that may be treated and/or prevented with the compounds and methods of the present invention include psoriasis, keloids and other hyperplasias that affect the skin, bullous disorders associated with epidermoid blister formation including bullous pemphigoid, erythema multiforme and dermatitis herpetiformis, fibrotic disorders such as pulmonary fibrosis, atherosclerosis, restenosis and cirrhosis, renal diseases including mesangial cell proliferative disorders, glomerulopathy, glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis and polycystic kidney disease, Benign Prostatic Hyperplasia (BPH), angiogenic or vasculoproliferative disorders, and thrombotic microangiopathy syndromes.

The compounds of the present invention are also useful in the treatment and/or prevention of ophthalmic diseases, for example, age-related macular degeneration (AMD), dry macular degeneration, ischemic retinal vein occlusion, diabetic macular edema, diabetic retinopathy, retinopathy of prematurity, and other retinopathies.

Other conditions that may be treated and/or prevented by administration of a compound of the invention include gynaecological diseases such as endometriosis, myoma and ovarian cysts, metabolic disorders associated with adipogenesis, bile metabolism, phosphate metabolism, calcium metabolism and/or bone mineralization, skeletal disorders such as, for example, dwarfism, chondroplastic disorders and Pfeiffer syndrome, cartilage diseases such as osteoarthritis and polyarthritis, rheumatoid arthritis, alopecia, and transplant rejection.

The diseases mentioned above have been well characterized in humans, but they also exist in other mammals with similar etiologies, and can be treated in those with the compounds and methods of the present invention.

The invention therefore also relates to the use of the compounds according to the invention for the treatment and/or prophylaxis of disorders, in particular of the disorders mentioned above.

The invention further relates to the use of the compounds according to the invention for the preparation of pharmaceutical compositions for the treatment and/or prophylaxis of disorders, in particular of the disorders mentioned above.

The invention further relates to the use of the compounds according to the invention in a method for the treatment and/or prophylaxis of disorders, in particular of the disorders mentioned above.

The present invention further relates to methods for the treatment and/or prevention of disorders, in particular of the above-mentioned disorders, by using an effective amount of at least one compound according to the invention.

The compounds of the present invention may be administered as a single agent or in combination with one or more other therapeutic agents, provided that such combination does not result in undesirable and/or unacceptable side effects. Such combination therapy includes administration of a single pharmaceutical dosage form containing a compound of formula (I) as described above and one or more other therapeutic agents, as well as administration of a compound of formula (I) and each other therapeutic agent in their respective separate pharmaceutical dosage forms. For example, the compound of formula (I) and the therapeutic agent may be administered to the patient together in a single (fixed) oral dosage composition, such as a tablet or capsule, or the individual agents may be administered in separate dosage formulations.

If separate dosage formulations are used, the compound of formula (I) and the one or more other therapeutic agents may be administered substantially simultaneously (i.e., simultaneously) or at separate staggered times (i.e., sequentially).

In particular, the compounds of the present invention may be used in fixed or separate combination with other anti-cancer agents, such as alkylating agents, antimetabolites, plant-derived anti-neoplastic agents, hormonal therapy agents, topoisomerase inhibitors, tubulin inhibitors, kinase inhibitors, targeting drugs, antibodies, antibody-drug conjugates (ADCs), immunological agents, biological response modifiers, anti-angiogenic compounds, and other anti-proliferative, cytostatic, and/or cytotoxic substances. In this regard, the following is a non-limiting list of examples of second agents that may be used in combination with the compounds of the present invention:

abarelix, abiraterone, aclarubicin, afatinib, aflibercept, aldesleukin, alemtuzumab, alitretinoin, alpharadin, altretamine, aminoglutethimide, amonafide, amrubicin, amsacrine, anastrozole, andramustine, arglabin, asparaginase, axitinib, 5-azacytidine, basiliximab, belotecan, bendamustine, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, bortezomib, bosutinib, alanine brimonib, buserelin, busulfan, cabazitaxel, CAL-101, calcium folinate, calcium levofolinate, camptothecin, capecitabine, carboplatin, carmoflavizumab, carmustine, cetuximab, cetrimide, bevacizumab, cetuximab, melphalan, chlorambucil, melphalan, meclizine, Cisplatin, cladribine, clodronic acid, clofarabine, combretastatin, crisantapase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, alfacapotine, darinapasin, dasatinib, daunomycin, decitabine, garelix, niinterleukin, denosumab, deslorelin, dibromospiro-ammonium chloride, docetaxel, doxetanib, doxifluridine, doxorubicin, dutasteride, eculizumab, efarouna, efaronine, eletric acid, etoricepamine, ilpatadine, endostatin, enocitabine, epirubicin, thidol, epothinol, epothilone alpha, epothilone beta, epothilone, eptaplatin, eprinobulin, erlotinib, estramustine, etoposide, vilitant, irinotecan, everolimus A, everolimus, efavirenz A, valsartan, doxetacin, doxorabicin, doxorabicine, doxorabic, Filgrastim, finasteride, velapine (flavopiridol), fludarabine, 5-fluorouracil, fluoxymesterone, flutamide, foretinib, formestane, fotemustine, fulvestrant, ganirelix, gefitinib, gemcitabine, gemtuzumab, gemmace, gemmoster, glufosimi, goserelin, histrelin, hydroxyurea, ibandronic acid, temozolomide, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, intedanib, interferon alpha-2 a, interferon alpha-2 b, interferon beta, interferon gamma, interleukin-2, prilomamab (ipilimumab), irinotecan, salsalaparin, lanreotide, lapatinib, lasifene, lexofenadine, grittin, polysaccharide, letrovaib, letrozole, levamisole, leuprolide, glimepiridib, glipizide, and other drugs, linifanib, linsitinib, lisuride, lobaplatin, lomustine, lonidamine, lurtotecan, macsfamide, mapatumumab, masitinib, maxolone, medroxyprogesterone, megestrol, melarsol, melphalan, meiandrane, mercaptopurine, methotrexate, methyl aminoacetonate, methyltestosterone, mivampitide, mifepristone, miltefosine, mibeplatin, dibromomannitol, mitoxantrone, dibromodulcitol, mitomycin, mitotane, mitoxantrone, moraxetin, motishanib, nandrolone, nedaplatin, nelarabine, neratinib, nilotinib, nilutamide, nimotuzumab, nimustine, diammonidine (nitrarine), lorartisone, olaratumab, oxepartin, oxaliplatin, paclitaxel, ritaminic acid, phospho, beta-furazalide, PEG-D-E, pegfilastrim, PEG-interferon alpha-2 b, pellitrexol, pemetrexed, pemtumumab, pentostatin, pellomycin, phosphoramide, perifosine, pertuzumab, streptolysin, pirambicin, pirarubicin, pratensefrin, plicamycin, chitosan, pratensamide, estradiol polyphosphate, panatine, porfimer sodium, pralatrexate, prednisolone, procarbazine, propiconazole, PX-866, quinagolide, raloxifene, raltitrexed, ranibizumab, ramustine, razoxan, regorafenib, risedronic acid, rituximab, romidepsin, rolimus, luminid, rubitecan, lucirinotecan, sainittin, sargrastim, setriplatin, simetidine, sipuuticuucel-T, cillebertib, zotocin, zornist, zornitussimus, temozolinitinib, sorafenib, temsirolimus, sorafenib, temustine, sorafenib, tasolinamine (tasonermin), teicoplanin, tegafur, tiratinib, temoporfin, temozolomide, temsirolimus, teniposide, testolactone, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thioguanine, tipifarnib, tivozanib, toceranib, tollizumab, topotecan, toremifene, tositumomab, trabectedin, trastuzumab, troosulfan, tretinoin, tripaine, trostan, tritrexate, triptorelin, trofosfamide (trosfamide), ubenix, valrubicin, vandetanib, varlitinib, valtararib, vemurafenib, vidarabine, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, voroxicib, voroxib, vorinosil, vorinostat, zorubicin, and valacilin.

In general, the following objectives can be sought with the compounds of the present invention in combination with other anti-cancer agents:

● improved activity in slowing tumor growth, reducing its size or even its complete elimination compared to treatment with the single active compound;

● the possibility of using a lower dose of chemotherapeutic agent than monotherapy;

● the possibility of a more tolerable therapy with fewer side effects than the individual administration;

● possibility of treating a broader spectrum of cancer and tumor diseases;

● achieve higher response rates to treatment;

● longer survival time of the patient compared to standard therapy.

Thus, in another embodiment, the present invention relates to a pharmaceutical composition comprising at least one compound of the present invention and one or more other therapeutic agents for the treatment and/or prevention of a disorder, in particular the disorders described above.

In the treatment of cancer, the compounds of the invention may also be employed together with radiotherapy and/or surgical intervention.

Further, the compounds of formula (I) can be used as such or in the form of compositions for research and diagnosis, or as analytical reference standards and the like, which are well known in the art.

When a compound of the invention is administered as a medicament to humans and other mammals, it may be administered by itself or as a pharmaceutical composition containing, for example, from 0.1% to 99.5% (more preferably, from 0.5% to 90%) of the active ingredient in combination with one or more pharmaceutically acceptable excipients.

Thus, in another aspect, the present invention relates to pharmaceutical compositions comprising at least one compound of the invention, and generally one or more inert, non-toxic, pharmaceutically suitable excipients, and their use for the treatment and/or prevention of disorders, in particular of the disorders mentioned above.

The compounds of the invention may act systemically and/or locally. For this purpose, it can be administered in a suitable manner, for example by oral, parenteral, pulmonary, nasal, lingual, sublingual, buccal, rectal, dermal, transdermal, conjunctival, aural or topical routes, or as an implant or stent.

For these administration routes, the compounds of the invention can be administered in a suitable administration form.

Suitable for oral administration are administration forms which function according to the prior art and deliver the compounds of the invention rapidly and/or in a modified manner and which contain the compounds of the invention in crystalline, amorphous and/or dissolved form, such as tablets (uncoated or coated tablets, for example with enteric coating or coatings which are insoluble or slow-dissolving and control the release of the compounds of the invention), tablets which disintegrate rapidly in the oral cavity, or films/wafers, films/lyophilisates, capsules (for example hard or soft gelatin capsules), sugar-coated tablets, granules, pills, powders, emulsions, suspensions, aerosols or solutions.

Parenteral administration can be carried out avoiding absorption steps (intravenous, intra-arterial, intracardiac, intraspinal or intralumbar), or involving absorption (intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal). Useful forms for parenteral administration include solutions, suspensions, emulsions, lyophilisates and injections and infusion preparations in the form of sterile powders.

Forms suitable for other routes of administration include, for example, inhaled pharmaceutical forms (e.g., powder inhalants, sprays), nasal drops, solutions or sprays, tablets or capsules for lingual, sublingual or buccal administration (e.g., troches, lozenges), suppositories, otic and ophthalmic preparations (e.g., drops, ointments), vaginal capsules, aqueous suspensions (lotions, shaking mixtures), lipophilic suspensions, ointments, creams (cream), milks (milk), pastes, foams, dusting powders, transdermal therapeutic systems (e.g., patches), implants and stents.

In a preferred embodiment, there is provided a pharmaceutical composition comprising a compound of formula (I) as defined above in a form suitable for oral administration. In another preferred embodiment, there is provided a pharmaceutical composition comprising a compound of formula (I) as defined above in a form suitable for intravenous administration.

The compounds of the invention can be converted into the administration forms described in a manner known per se by mixing with inert, nontoxic, pharmaceutically suitable excipients. These excipients include, inter alia, carriers (e.g., microcrystalline cellulose, lactose, mannitol), solvents (e.g., liquid polyethylene glycol), emulsifiers (e.g., sodium lauryl sulfate), surfactants (e.g., polyoxysorbitan oleate), dispersants (e.g., polyvinylpyrrolidone), synthetic and natural polymers (e.g., albumin), stabilizers (e.g., antioxidants such as ascorbic acid), colorants (e.g., inorganic pigments such as iron oxides), and flavoring and/or odor masking agents.

The preferred dosage of the compounds of the invention is the maximum that can be tolerated by the patient without serious side effects. For example, the compounds of the present invention may be administered parenterally at a dosage of about 0.001 mg/kg to about 1 mg/kg, preferably about 0.01 mg/kg to about 0.5 mg/kg body weight. For oral administration, exemplary dosages range from about 0.01 to 100 mg/kg, preferably from about 0.01 to 20 mg/kg, and more preferably from about 0.1 to 10 mg/kg of body weight. Ranges intermediate to the above values are also intended to be part of the present invention.

However, the actual dosage level and timing of administration of the active ingredient in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. Accordingly, it may be necessary to deviate from the amounts indicated, especially with respect to age, sex, body weight, diet and general health of the patient, the bioavailability and pharmacokinetic characteristics of the particular compound and its mode and route of administration, the time or interval at which administration occurs, the dosage regimen selected, the response of the individual patient to the active ingredient, the particular disease involved, the extent or involvement or severity of the disease, the nature of concurrent therapy (i.e., the interaction of the compound of the invention with other co-administered therapeutic agents), and other relevant circumstances.

Thus, in some cases it may be satisfactory to treat with amounts lower than the minimum amounts mentioned above, while in other cases the upper limit must be exceeded. Treatment can be initiated using smaller doses than the optimal dose of the compound. The dose can then be increased in small increments until the optimum effect under the circumstances is achieved. For convenience, the total daily dose may be divided and administered in portions interspersed throughout the day.

The following exemplary embodiments illustrate the invention. The present invention is not limited to these examples.

Unless otherwise indicated, percentages in the following tests and examples are by weight; parts are by weight. The reported solvent ratio, dilution ratio and concentration of the liquid/liquid solution are each by volume.

A.Examples

Abbreviations and acronyms:

ac acetyl group

Ac₂O acetic anhydride

AcOH acetic acid

aq. Water (solution)

Boc tert-butoxycarbonyl

br. wide (¹H-NMR Signal)

Bu butyl

cat, catalyzed

conc. concentrated

d doublet of peaks (¹H-NMR Signal)

DBDMH 1, 3-dibromo-5, 5-dimethylhydantoin

DCI direct chemical ionization (MS)

DCM dichloromethane

Dess-martin oxidant 1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3 (1)H) -ketones

DIPEAN,N-diisopropylethylamine

DMFN,N-dimethylformamide

DMSO dimethyl sulfoxide

EI Electron bombardment ionization (MS)

eq. equivalent

ESI electrospray ionization (MS)

Et Ethyl group

EtOAc ethyl acetate

GC-MS gas chromatography combined mass spectrum

h hours

Hal halogen

¹H-NMR proton nuclear magnetic resonance spectrum method

HPLC high performance liquid chromatography

iPr isopropyl group

LC-MS liquid chromatography combined mass spectrum

Me methyl group

MeOH methanol

min for

MS mass spectrometry

m/z Mass to Charge ratio (MS)

NBSN-bromosuccinimide

n-Bu n-butyl

NCSN-chlorosuccinimide

of th. theory (chemical yield)

Pd/C carbon-supported palladium

PdCl₂(dppf) [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (II)

Pd(dba)₂Bis (dibenzylideneacetone) palladium

Ph phenyl

PPA polyphosphoric acid

q quartet: (¹H-NMR Signal)

quantitive (yield)

rac racemization

R_fTLC retention factor

RP reverse phase (HPLC)

rt Room temperature

R_tRetention time (HPLC)

s singlet (¹H-NMR Signal)

sat. saturation (solution)

t triplet (¹H-NMR Signal)

TBAF tetra-n-butylammonium fluoride

TBDMS tert-butyldimethylsilyl group

TBTUN-[(1H-benzotriazol-1-yloxy) (dimethylamino) methylene]-N-methyltetrafluoro

Methylammonium borate (methanaminium tetrahluoroorate)

tBu tert-butyl

tertTertiary amine

TFA trifluoroacetic acid

THF tetrahydrofuran

TLC thin layer chromatography.

LC-MS and GC-MS methods:

method 1 (LC-MS):

the instrument comprises the following steps: micromass Quattro premix with Waters UPLC Acquity; column: ThermoHypersil GOLD 1.9 μm, 50 mm x1 mm; eluent A: 1L of water +0.5 mL of 50% aqueous formic acid, eluent B: 1L acetonitrile +0.5 ml 50% aqueous formic acid; gradient: 0.0 min 90% A → 0.1 min 90% A → 1.5 min10% A → 2.2 min10% A; temperature: 50 ℃; flow rate: 0.33 mL/min; and (4) UV detection: 210 nm.

Method 2 (LC-MS):

the instrument comprises the following steps: waters Acquity SQD UPLC System; column: waters Acquity UPLC HSS T31.8 mu m, 50 mm x1 mm; eluent A: 1L of water + 0.25 mL of 99% formic acid, eluent B: 1L acetonitrile + 0.25 mL99% formic acid; gradient: 0.0 min 90% A → 1.2 min5% A → 2.0 min5% A; furnace temperature: 50 ℃; flow rate: 0.40 mL/min; and (4) UV detection: 210-400 nm.

Method 3 (LC-MS):

the instrument comprises the following steps: micromass Quattro Micro with HPLC Agilent 1100 series; column: YMC-TriartC 183 mu m, 50 mm x 3 mm; eluent A: 1L of water + 0.01 mol of ammonium carbonate, eluent B: 1L of acetonitrile; gradient: 0.0 min 100% A → 2.75 min5% A → 4.5 min5% A; furnace temperature: 40 ℃; flow rate: 1.25 mL/min; and (4) UV detection: 210 nm.

Method 4 (LC-MS):

the instrument comprises the following steps: waters Acquity SQD UPLC System; column: waters Acquity UPLC HSS T31.8 mu m, 30 mm x 2 mm; eluent A: 1L of water + 0.25 mL of 99% formic acid, eluent B: 1L acetonitrile + 0.25 mL99% formic acid; gradient: 0.0 min 90% A → 1.2 min5% A → 2.0 min5% A; furnace temperature: 50 ℃; flow rate: 0.60 mL/min; and (4) UV detection: 208-400 nm.

Method 5 (LC-MS):

the instrument comprises the following steps: micromass Quattro Premier with Waters UPLC Acquity; column: ThermoHypersil GOLD 1.9 μm, 50 mm x1 mm; eluent A: 1L of water +0.5 mL of 50% aqueous formic acid, eluent B: 1L acetonitrile +0.5 ml 50% aqueous formic acid; gradient: 0.0 min 97% A → 0.5 min 97% A → 3.2 min5% A → 4.0 min5% A; temperature: 50 ℃; flow rate: 0.3 mL/min; and (4) UV detection: 210 nm.

Method 6 (GC-MS):

the instrument comprises the following steps: micromass GCT, GC 6890; column: restek RTX-35, 15m x 200 μm m x 0.33.33 μm; helium constant flow: 0.88 mL/min; furnace temperature: 70 ℃; a sample inlet: 250 ℃; gradient: 70 ℃,30 ℃/min → 310 ℃ (hold for 3 min).

Method 7 (LC-MS):

and (4) an instrument MS: waters Micromass QM; instrumental HPLC, Agilent 1100 series; column: AgilentZORBAX extended-C183.0 mm x 50 mm, 3.5 mu m; eluent A: 1L of water + 0.01 mol of ammonium carbonate, eluent B: 1L of acetonitrile; gradient: 0.0 min 98% A → 0.2 min 98% A → 3.0 min5% A → 4.5 min5% A; temperature: 40 ℃; flow rate: 1.75 mL/min; and (4) UV detection: 210 nm.

Method 8 (LC-MS):

and (4) an instrument MS: waters Micromass ZQ; instrumental HPLC, Agilent 1100 series; column: AgilentZORBAX extended-C183.0 mm x 50 mm, 3.5 mu m; eluent A: 1L of water + 0.01 mol of ammonium carbonate, eluent B: 1L of acetonitrile; gradient: 0.0 min 98% A → 0.2 min 98% A → 3.0 min5% A → 4.5 min5% A; temperature: 40 ℃; flow rate: 1.75 mL/min; and (4) UV detection: 210 nm.

General purification method(see tables I and II below):

Method P1:

preparative RP-HPLC (Reprosil C18, gradient acetonitrile/0.2% aqueous trifluoroacetic acid).

Method P2:

preparative RP-HPLC (XBridge C18, gradient acetonitrile/water +0.1% ammonia).

Method P3:

preparative RP-HPLC (Sunfire C18, gradient acetonitrile/water).

Method P4:

preparative RP-HPLC (XBridge C18, gradient acetonitrile/water + 0.05% ammonia).

Method P5:

the product obtained from the above RP-HPLC purification was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column was eluted with methanol and the filtrate was evaporated.

Method P6:

the product solution in ethyl acetate was washed with saturated aqueous sodium bicarbonate solution followed by saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered and evaporated.

Starting materials and intermediates:

intermediate 1A

2-methoxy-4-methylaniline

A mixture of 5-methyl-2-nitroanisole (265 g, 1.58 mol) and 10% Pd/C (39.75 g) in THF (1.32L) was stirred overnight at room temperature under 1 atm hydrogen. Filtration through celite and evaporation gave 216.1 g of crude product which was used in the next step without further purification.

Intermediate 2A

2-methoxy-4-methylthiophenol

。

The method comprises the following steps:

a solution of sodium nitrite (7 g, 101.4 mmol) in water (25 ml) was added dropwise to a cooled (0 ℃ C. -5 ℃ C.) solution of intermediate 1A (13.7 g, 100 mmol) in concentrated hydrochloric acid (30 ml) and water (85 ml). After stirring at 0 ℃ for 10 min, sodium acetate (15 g, 182.8 mmol) was added. The resulting mixture was added dropwise to a hot solution (70 ℃ -80 ℃) of o-ethyldithiopotassium carbonate (30 g, 187.1 mmol) in water (140 ml), stirred between 70 ℃ and 80 ℃ for 1h, then cooled to room temperature. The mixture was extracted twice with ethyl acetate and the combined organic extracts were dried over sodium sulfate and evaporated. The residue was dissolved in a 1.3M solution of potassium hydroxide in ethanol (300 ml). Glucose (8g) was added and the resulting mixture was refluxed for 3 h. Then, the ethanol solvent was evaporated, and the residue was diluted with water and acidified with 6N aqueous sulfuric acid. Zinc powder (15 g) was carefully added and the resulting mixture was heated to 50 ℃ for 30 min. The mixture was then cooled to room temperature, diluted with dichloromethane and filtered. The filtrate was extracted twice with dichloromethane and the combined organic extracts were dried over sodium sulfate and evaporated to give 14.3 g of crude product which was used in the next step without further purification.

The method 2 comprises the following steps:

to 2.9L THF was added 355 ml (6.67 mol) of a warm solution of concentrated sulfuric acid in 1.1L water. 293 g (1.33 mol) of 2-methoxy-4-methylbenzenesulfonyl chloride are added at 50 ℃ with stirring. Then 521g (7.97 mol) zinc powder (foaming) was carefully added in portions and the slightly exothermic reaction was cooled in a water bath to maintain a temperature of 50-55 ℃. The mixture was then stirred at 55 ℃ for 3 h. The progress of the reaction was monitored by TLC (silica gel, petroleum ether/ethyl acetate 95: 5). The reaction mixture was poured into 13.6L of water, 6.8L of dichloromethane was added, and the mixture was stirred for 5 min. After decantation from the remaining zinc and phase separation, the aqueous phase is extracted once more with 6.8L of dichloromethane. The combined organic phases were washed with 10% brine, dried and evaporated at 40 ℃ under reduced pressure to give 237 g of crude product. This material was used in the next step without further purification. Analytical samples were obtained by silica gel chromatography using petroleum ether/ethyl acetate (97:3) as eluent.

Intermediate 3A

1- [ (2, 2-diethoxyethyl) thio ] -2-methoxy-4-methylbenzene

237 g of the crude material from intermediate 2A, 287 g (1.46 mol) of bromoacetaldehyde diethyl acetal and 862 g (2.65 mol) of cesium carbonate were suspended in 2L of DMF. The reaction temperature was initially increased to 40 ℃ and stirring was then continued overnight at room temperature. The reaction mixture was partitioned between 10L water and 2.7L ethyl acetate. The aqueous phase is extracted with another 2.7L of ethyl acetate. The combined organic phases were washed with 10% brine, dried and evaporated. The oily residue obtained is purified by chromatography on silica gel using petroleum ether/ethyl acetate (95:5) as eluent.

Yield: 236 g of an oil (66% of theory).

Intermediate 4A

7-methoxy-5-methyl-1-benzothiophene

To a refluxing mixture of 13 g polyphosphoric acid and 150 ml chlorobenzene was added dropwise a solution of 5.2 g (19.2 mmol) of intermediate 3A and the reflux was continued overnight. After cooling, the organic layer was decanted, and the residue and flask were rinsed twice with DCM. The combined organic phases were evaporated under reduced pressure. The residue (3.76 g) was chromatographed on silica gel using isohexane/0-10% ethyl acetate as eluent.

Yield: 1.69 g of an oil (49% of theory).

Intermediate 5A

(7-methoxy-5-methyl-1-benzothien-2-yl) boronic acid

26.7 g (150 mmol) of intermediate 4A were dissolved in 270 ml THF under an argon atmosphere and cooled to-70 ℃.66 ml (165 mmol) of a 2.5N solution of N-butyllithium in hexane were added dropwise at between-70 ℃ and-65 ℃ over 20min, resulting in the formation of a white precipitate. After stirring at-70 ℃ for 1h, 41.5 ml (180 mmol) of triisopropyl borate are added at this temperature over 10 min (resulting in a thick suspension). Stirring was continued at-70 ℃ for 1h, then the reaction mixture was allowed to warm to room temperature overnight. Then, 400 ml of a saturated aqueous ammonium chloride solution was added, the layers were separated, and the aqueous layer was extracted once more with THF. The combined organic phases were evaporated under reduced pressure. To the residue thus obtained were added 200 ml of water and 86 ml of a 2N aqueous sodium hydroxide solution. The solution was washed twice with DCM and then acidified with 35 ml 3M sulphuric acid and the resulting suspension was stirred vigorously for 1 h. The precipitate was filtered off with suction and dried overnight under vacuum at 45 ℃.

Yield: 28.25 g of a colourless solid (purity 94% by LC-MS, 80% of theory).

Intermediate 6A

2- (7-methoxy-5-methyl-1-benzothien-2-yl) -6-methyl-1, 3,6, 2-dioxazaborolan-4, 8-dione

6.3 g (28.4 mmol) of intermediate 5A and 4.2 g (28.4 mmol) of 2,2' - (methylimino) diacetic acid are dissolved in a mixture of 45 ml of DMSO and 400 ml of toluene and refluxed for 16 h using a Dean-Stark trap. After evaporation, the residue was dissolved in ethyl acetate and washed three times with water and once with brine. The organic phase was dried over magnesium sulfate and evaporated to a volume of about 200 ml. A white solid precipitated, which was filtered, washed with ethyl acetate and dried in vacuo to give the first crop of the title compound (5.52 g). The mother liquor was evaporated and subjected to flash chromatography on silica gel using cyclohexane/0-100% ethyl acetate as eluent to obtain a second crop (3.32 g).

Yield: 8.84 g (total purity by LC-MS 92.5%, 87% of theory).

Intermediate 7A

4-Aminopyrrolo [2,1-f ] [1,2,4] triazine-6-carboxylic acid ethyl ester

A solution of 4-aminopyrrolo [2,1-f ] [1,2,4] triazine-6-carbonitrile (3.9 g, 24.5 mmol; preparation described in PCT International patent application WO 2007/064883) in ethanol (124.8 ml) was stirred with concentrated sulfuric acid (62.4 ml) at 80 ℃ overnight. After cooling to room temperature, the reaction mixture was poured onto 800 g of ice and adjusted to pH 6-7 with concentrated aqueous sodium hydroxide solution. Ethyl acetate (500 ml) and dichloromethane (500 ml) were added to the suspension and the resulting mixture was filtered through celite. The organic layer was separated from the aqueous layer. The solid was dissolved in hot water (1L) and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were dried over sodium sulfate and evaporated. The residue is triturated with an isopropanol/diethyl ether mixture and the solid is filtered off to yield 2.5 g (49% of theory) of the title compound.

Intermediate 8A

(4-Aminopyrrolo [2,1-f ] [1,2,4] triazin-6-yl) methanol

An ice-cold solution of intermediate 7A (3.0 g, 14.5 mmol) in THF (30 ml) was treated with a 1M solution of lithium triethylborohydride in THF (58 ml) and stirred at rt for 45 min. The reaction mixture was then cooled to 0 ℃, quenched with methanol, slowly warmed to room temperature and adsorbed onto celite. Purification by column chromatography on silica gel (dichloromethane/methanol 20:1 → 4:1 gradient) gives 2.21 g (92.5% of theory) of the title compound.

Intermediate 9A

[ 2-cyano-4- (hydroxymethyl) -1H-pyrrol-1-yl]Carbamic acid tert-butyl ester

A1M solution of methylmagnesium bromide in THF (13.3 ml) was added over 15min to a solution of tert-butyl (4-bromo-2-cyano-1H-pyrrol-1-yl) carbamate (3.7 g, 12.09 mmol; preparation described in PCT International patent application WO2007/064883, intermediate AAE, step 3) in THF (37 ml) cooled to-60 ℃ under argon. After 30min, a 1.6M solution of n-butyllithium in hexane (15.1 ml, 24.2 mmol) was added to the reaction over 10 min, and the reaction mixture was stirred between-60 ℃ and-40 ℃ for 1 h. Then, paraformaldehyde (1.09 g, 36.3 mmol) was added to the reaction, and the reaction mixture was slowly warmed to room temperature and stirred overnight. After quenching with saturated aqueous ammonium chloride solution, the aqueous layer was extracted twice with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate and evaporated. Purification by chromatography on silica gel (cyclohexane/ethyl acetate 2:1 → 1:1) gives 2.04 g (69% of theory) of the title compound.

Intermediate 10A

6- (methoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

。

The method comprises the following steps:

a solution of intermediate 8A (1.3 g, 7.9 mmol) in THF (25 ml) was treated with thionyl chloride (1.15 ml,15.8 mmol) and stirred at room temperature for 2 h. After evaporation, the residue was dissolved in methanol (25 ml) and treated with sodium acetate (1.3 g, 15.8 mmol). The mixture was stirred at 65 ℃ for 3h and then evaporated again. Purification by column chromatography on silica gel (dichloromethane/methanol 100:2) gives 787 mg (55% of theory) of the title compound.

The method 2 comprises the following steps:

a solution of intermediate 9A (6.14 g, 25.88 mmol) in a 4M solution of hydrochloric acid/1, 4-dioxane (15 ml) was stirred at room temperature for 5 h. After dilution with methanol (73 ml), stirring was continued overnight at room temperature. Then, potassium phosphate (54.9 g, 258.65 mmol) and formamidine acetate (13.46 g, 129.32 mmol) were added, and the resulting mixture was stirred at 65 ℃ for 17 h. The reaction mixture was evaporated, saturated aqueous sodium chloride solution was added, and the mixture was extracted with dichloromethane, followed by extraction with ethyl acetate. The combined organic phases were dried over sodium sulfate and evaporated. Purification by column chromatography on silica gel (dichloromethane/methanol 40:1 → 20:1) gives 2.36 g (49% of theory) of the title compound.

Intermediate 11A

4-amino-6- (methoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

Phosphorus oxychloride (13.7 ml, 147.18 mmol) was added to a solution of intermediate 10A (5.24 g, 29.43mmol) in DMF (80 ml) at 0 ℃. The reaction mixture was stirred at 60 ℃ for 8 h, then carefully quenched with water and neutralized with 4M aqueous sodium hydroxide. The aqueous layer was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated. A solution of the residue in methanol (50 ml) was treated with sodium acetate (2.41 g, 29.43mmol) and refluxed overnight. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to give 2.66 g of crude product, which was used in the next step without further purification.

Intermediate 12A

4-amino-5-bromo-6- (methoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

A solution of intermediate 11A (crude, 2.66 g) in DMF (30 ml) cooled to-30 ℃ was taken upNTreatment of a solution of bromosuccinimide (NBS; 2.52 g, 14.19 mmol) in DMF (14 ml). The resulting mixture was slowly warmed to 0 ℃. After 1 hour, the mixture was warmed to room temperature, stirred for a further 15min, and then quenched with 1M aqueous sodium thiosulfate. The precipitate was filtered off and washed with ethyl acetate to yield 1.1 g (100% purity, 30% of theory) of the title compound as a first crop. The remaining filtrate was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate and evaporated. The residue was purified by silica gel column chromatography (cyclohexane/ethyl acetate 1:1 → 1:3) to further obtain 1.39 g (70% pure)Degree, 26% of theory) of the title compound.

Intermediate 13A

4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

Degassed 0.5M aqueous potassium phosphate solution (9.9 ml) was added to intermediate 12A (710 mg,2.49 mmol), intermediate 5A (921 mg, 3.73 mmol) and (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; 196 mg, 249 μmol; see S.L. Buchwald et al, under argon, J. Am. Chem. Soc.132 (40) 14073-14075(2010)) in degassed THF (28.4 ml). The resulting mixture was stirred at 60 ℃ for 2h and then evaporated. The residue is purified by column chromatography on silica gel (cyclohexane/ethyl acetate 5:1 → 1:1) to yield 550 mg (51% of theory) of the title compound.

Intermediate 14A

6- (ethoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

。

The method comprises the following steps:

to a solution of 2 g (12.2 mmol) of intermediate 8A in 40 ml THF was added 1.78 ml (24.4 mmol) of thionyl chloride at room temperature over 20 seconds. The mixture was stirred for 1.5 h, then evaporated to dryness and the residue was dissolved in 40 ml ethanol. 2 g (24.4 mmol) of sodium acetate are added and the mixture is stirred at 70 ℃ for 1h 45 min. The reaction mixture was evaporated again and saturated aqueous sodium bicarbonate solution was added. The mixture was extracted five times with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to dryness to give 2.02 g of crude product which was purified by column chromatography on silica gel using dichloromethane/methanol (0-2%) as eluent.

Yield: 1.37 g (58% of theory).

The method 2 comprises the following steps:

step 1: a solution of intermediate 9A (2.3 g, 9.69 mmol) in1, 4-dioxane (5ml) was treated with a 4M solution of hydrochloric acid in1, 4-dioxane (24 ml, 96.9 mmol) and stirred at room temperature for 130 min. The suspension is then filtered and the precipitate is washed with 1, 4-dioxane (5ml) and dried in vacuo to give 1.01 g (54% of theory) of the intermediate compound 1-amino-4- (chloromethyl) -1H-pyrrole-2-carbonitrile hydrochloride.

Step 2: freshly prepared 1-amino-4- (chloromethyl) -1 from step 1H-pyrrole-2-carbonitrile hydrochloride (0.3g, 1.82 mmol) was dissolved in ethanol (10 ml) and stirred at room temperature for 5 min. The clear solution was treated with formamidine acetate (813mg, 7.81 mmol) and potassium phosphate (1.66 g, 7.81 mmol) and stirred first at room temperature for 3 days and then at 80 ℃ for 10.5 h. More formamidine acetate (488 mg, 4.69 mmol) was added and the mixture was stirred at 80 ℃ for a further 18 h. The mixture was then cooled to room temperature and water and ethyl acetate were added. The organic phase was separated and the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated. The residue is dissolved in a mixture of methanol and dichloromethane, adsorbed onto kieselguhr, dried in vacuo and finally purified by chromatography on silica gel (gradient 0-10% methanol in dichloromethane) to give 260 mg (78% of theory) of the title compound。

Intermediate 15A

4-amino-6- (ethoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

To a solution of 2.1 g (10.9 mmol) of intermediate 14A in 40 ml of dry DMF at 0 ℃ under an argon atmosphere was added dropwise 5.1 ml (54.6 mmol) of phosphorus oxychloride. The mixture was stirred at 60 ℃ for 10 h. Water was then added carefully and the mixture was stirred at room temperature until all reaction intermediates were destroyed (HPLC control). The acid solution was neutralized with 1M aqueous sodium hydroxide solution and extracted three times with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure.

Yield: 1.94 g (purity 90%, 81% of theory).

Intermediate 16A

4-amino-5-bromo-6- (ethoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

To a solution of 73 g (0.33 mol) of intermediate 15A in 1.9L DMF at-15 deg.C was added dropwise a solution of 65 g (0.37 mol) NBS in 200 ml DMF. The mixture was warmed to 0 ℃ and stirred at this temperature for 3 h. The reaction mixture was poured into 2% aqueous sodium thiosulfate solution with stirring, and the precipitate was filtered off, washed with water and dried over phosphorus pentoxide in vacuo.

Yield: 85.6 g (86% of theory).

Intermediate 17A

4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

714 mg (85% purity, 2.03 mmol) of intermediate 16A, 946 mg (2.84mmol) of intermediate 6A and 160 mg (0.2 mmol) (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; see S.L. Buchwald et al,J. Am. Chem. Soc.132 (40) 14073-14075(2010)) was suspended in 15.5 ml of THF. Then, 15.5 ml of degassed 0.5M aqueous potassium phosphate solution was added, and the mixture was stirred at 50 ℃ for 16 h. After addition of water, the mixture was extracted with ethyl acetate and the combined organic phases were dried over magnesium sulfate and evaporated under reduced pressure. The residue was purified by column chromatography on silica gel (100 g) using 10-50% ethyl acetate/cyclohexane as eluent.

Yield: 452 mg (75% pure by HPLC, 42% of theory).

Intermediate 18A

(4-amino-5, 7-dibromopyrrolo [2,1-f ] [1,2,4] triazin-6-yl) methanol

A solution of intermediate 8A (5 g, 30.4 mmol) in THF (100 ml) was treated with 1, 3-dibromo-5, 5-dimethylhydantoin (9.58 g, 33.5 mmol) and stirred at room temperature for 2 h. The precipitate is filtered off and dried in vacuo to yield 6.60 g (64% of theory) of the title compound.

Intermediate 19A

(4-amino-5-bromopyrrolo [2,1-f ] [1,2,4] triazin-6-yl) methanol

A suspension of intermediate 18A (3.7 g, 11.5 mmol) in THF (800 ml) was heated with stirring until complete dissolution. The mixture was then cooled to-78 ℃ and a 1.6M solution of n-butyllithium in hexane (20ml, 32.1mmol) was added dropwise. After 5min, another 1.6M n-butyllithium solution (1.5 ml, 2.29mmol) was added. The resulting mixture was stirred at-78 ℃ for 5min, then quenched with methanol (5ml) and warmed to room temperature. The reaction mixture was diluted with saturated aqueous ammonium chloride solution, saturated aqueous sodium bicarbonate solution, saturated aqueous sodium chloride solution and ethyl acetate. After phase separation, the organic layer was washed with saturated aqueous sodium chloride solution. The combined aqueous phases were extracted again with ethyl acetate. The combined organic phases were washed again with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to give 2.87 g of crude product, which was used in the subsequent step without further purification.

Intermediate 20A

[ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methanol

A suspension of intermediate 19A (70% purity, 2.52 g, 7.26 mmol), intermediate 6A (3.63 g,10.9 mmol) and cesium fluoride (5.51 g, 36.3 mmol) in a THF/water mixture (10:1; 80 ml) was degassed under argon. Addition of 4- (di-tert-butylphosphino) -N,NDimethylaniline-palladium dichloride (2:1; 176 mg, 0.248 mmol) and the resulting mixture was degassed again and stirred at 50 ℃ for 16 h. The reaction mixture was then washed with saturated aqueous sodium chloride solution and the organic layer was separated, dried over magnesium sulfate, filtered and evaporated. The residue is suspended in methanol and the resulting solid is filtered off and dried in vacuo to yield 1.97 g (90% purity, 72% of theory) of the title compound.

Intermediate 21A

6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of intermediate 20A (400 mg, 1.17 mmol) in dichloromethane (12 ml) was treated with thionyl chloride (128. mu.l, 1.76 mmol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in methanol (12 ml) and treated with DIPEA (409. mu.l, 2.35 mmol). The mixture was refluxed overnight and then evaporated again. Purification by column chromatography on silica gel (dichloromethane/methanol 98:2 → 95:5) gives 388 mg (93% of theory) of the title compound.

Intermediate 22A

6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

Intermediate 20A (200 mg, 587. mu. mol) in dichloromethane (5ml) was treated with thionyl chloride (64. mu.l, 881. mu. mol) and stirred at room temperature for 15 min. After evaporation, the residue was refluxed in ethanol (5ml) for 1h, then treated with DIPEA (204. mu.l, 1.17 mmol) and refluxed again overnight. The reaction mixture is evaporated and the crude product is purified by column chromatography on silica gel (dichloromethane/methanol 98:2 → 95:5) to yield 202 mg (90% of theory) of the title compound.

Intermediate 23A

4- { [ 4-amino-6- (hydroxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazine-1-carboxylic acid tert-butyl ester

A solution of intermediate 20A (9.5 g, 27.9 mmol) in acetic acid (136.8 ml) was treated with tert-butyl piperazine-1-carboxylate (6.24 g, 33.49 mmol) and 37% aqueous formaldehyde (2.5 ml, 33.49 mmol). The mixture was stirred at 60 ℃ for 2.5 h. After evaporation, the residue was dissolved in ethyl acetate and the mixture was washed twice with saturated aqueous sodium bicarbonate, dried over sodium sulfate and evaporated. The residue was triturated in boiling ethanol (100 ml). The solid is filtered off and washed with ethanol and diethyl ether to yield 9.70 g (58% of theory) of the title compound.

Intermediate 24A

4- ({ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6- [ (2-methoxy-2-oxoethoxy) methyl ] pyrrolo [2,1-f ] [1,2,4] triazin-7-yl } methyl) piperazine-1-carboxylic acid tert-butyl ester

A solution of intermediate 23A (300 mg, 556. mu. mol) in dichloromethane (12 ml) was treated with thionyl chloride (81. mu.l, 1.11 mmol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in freshly distilled methyl glycolate (2.5 ml) and treated with DIPEA (485. mu.l, 2.78 mmol). The mixture was stirred at 70 ℃ for 2h, then evaporated again and the excess methyl glycolate removed by distillation. The residue is purified by column chromatography on silica gel (cyclohexane/ethyl acetate 1:5) to yield 136 mg (33% of theory) of the title compound.

Intermediate 25A

4- { [ 4-amino-6-formyl-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazine-1-carboxylic acid tert-butyl ester

A solution of intermediate 23A (300 mg, 556. mu. mol) in dichloromethane (4.2 ml) was treated with dess-martin oxidant (1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1, 1-benzidoxol)H) -ketone 307 mg, 724. mu. mol) and stirred at room temperature for 2 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate and saturated aqueous sodium thiosulfate (1:1) and stirred at room temperature for 30 min. The aqueous layer was extracted three times with dichloromethane and the combined organic layers were dried over magnesium sulfate and evaporated. Purification by column chromatography on silica gel (cyclohexane/ethyl acetate 1:1 → 100% ethyl acetate) gives 273 mg (87% of theory) of the title compound.

Intermediate 26A

4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazine-6-carbaldehyde di (formate)

A solution of intermediate 23A (80 mg, 148. mu. mol) in THF (3.9 ml) was treated with dess-martin oxidant (1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1, 1-benzidoxol)H) 94 mg, 222. mu. mol) and stirred at room temperature for 30 min. The reaction mixture was quenched with saturated aqueous sodium bicarbonate and saturated aqueous sodium thiosulfate (1: 1). The aqueous layer was extracted three times with ethyl acetate and the combined organic layers were dried over magnesium sulfate and evaporated. The residue was dissolved in 4M solution of hydrochloric acid in1, 4-dioxane (4ml) and stirred at room temperature for 1 h. After evaporation, the residue was passed through preparative RP-HPLC (Reprosil)C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 34mg (52% of theory) of the title compound.

Intermediate 27A

4- ({ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6- [ (3-oxopiperazin-1-yl) methyl ] pyrrolo [2,1-f ] [1,2,4] triazin-7-yl } methyl) piperazine-1-carboxylic acid tert-butyl ester

A solution of intermediate 25A (185 mg, 344. mu. mol) in THF (4.6 ml) was treated with 2-oxopiperazine (344mg, 3.4 mmol), sodium triacetoxyborohydride (365 mg, 1.7 mmol) and acetic acid (39. mu.l, 689. mu. mol). The resulting mixture was stirred at room temperature for 2h, then adsorbed onto celite and purified by silica gel column chromatography (dichloromethane → dichloromethane/methanol 100:8) to give 221 mg (quantitative) of the title compound.

Intermediate 28A

4-amino-7- { [4- (tert-butoxycarbonyl) piperazin-1-yl]Carbonyl } -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazine-6-carboxylic acidN-ethyl ammonium salt

A solution of intermediate 25A (70 mg, 130. mu. mol) in THF/water (10:1, 4.85 ml) was treated with a 2M solution of 2-methyl-2-butene in THF (521. mu.l, 1.04 mmol) and sodium dihydrogen phosphate (107 mg, 783. mu. mol) and stirred at room temperature for 5 min. Sodium chlorite (70 mg, 783 μmol) was added, and the resulting mixture was stirred at room temperature overnight. After dilution with water, the aqueous phase is extracted three times with ethyl acetate. The combined organic layers were dried over sodium sulfate and evaporated, and the residue was purified by preparative RP-HPLC (XBridge C18, gradient 5-50% acetonitrile/water + 0.05% diethylamine) to yield 18mg (21% of theory) of the title compound.

Intermediate 29A

4- { [ 4-amino-6- (azidomethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazine-1-carboxylic acid tert-butyl ester

A solution of intermediate 23A (150 mg, 0.278 mmol) in dichloromethane (7.5 ml) was treated with thionyl chloride (40 μ l, 0.56 mmol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in DMF (6 ml) and treated with sodium azide (362 mg, 5.57 mmol) and sodium iodide (208 mg,1.39 mmol). The mixture was heated to 80 ℃ for 1h, then diluted with water and extracted three times with ethyl acetate. The combined organic layers were washed with water and with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated. Purification by column chromatography on silica gel (cyclohexane/ethyl acetate 98:2 → 100% ethyl acetate) gives 95.8 mg (57% of theory) of the title compound.

Intermediate 30A

4- { [6- (Acetylaminomethyl) -4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazine-1-carboxylic acid tert-butyl ester

A mixture of intermediate 29A (320 mg, 567. mu. mol), 10% Pd/C (320 mg) and acetic anhydride (106. mu.l, 1.13 mmol) in methanol (32 ml) under 1 atm hydrogen was stirred at room temperature for 90 min. The mixture was then filtered through celite and the filtrate was evaporated. Purification by silica gel column chromatography (cyclohexane/ethyl acetate 1:1 → 100% ethyl acetate) gave 440 mg (quantitative) of the title compound.

Intermediate 31A

7- [ (4-acetylpiperazin-1-yl) methyl ] -4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-6-carbaldehyde

A solution of example 55(678 mg, purity 89%, 1.26 mmol) in dichloromethane (4ml) containing molecular sieve (4A) was treated with Dass-Martin oxidant (1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1H) -one; 623 mg, 1.47 mmol) and stirred at room temperature for 5 min. The reaction mixture is then adsorbed onto celite and purified by column chromatography on silica gel (30-100% ethyl acetate/cyclohexane gradient, then 0-10% methanol/dichloromethane) to give 449 mg (49% of theory) of the title compound.

Intermediate 32A

4- { [ 4-amino-6- (azidomethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

A solution of example 13(59 mg, 130. mu. mol) in dichloromethane (3.5 ml) was treated with thionyl chloride (19. mu.l, 261. mu. mol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in DMF (2.8 ml) and treated with sodium iodide (97 mg, 652. mu. mol) and sodium azide (169 mg, 2.6 mmol). The mixture was stirred at 80 ℃ for 1 h. After dilution with saturated aqueous sodium chloride solution, the aqueous phase was extracted four times with ethyl acetate and the combined organic layers were dried over magnesium sulfate and evaporated. Purification by column chromatography on silica gel (dichloromethane/methanol 98:2 → 90:10) gives 35 mg (56% of theory) of the title compound.

Intermediate 33A

4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (morpholin-4-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazine-6-carbaldehyde

A solution of example 50(710 mg, 70% purity, 1.13 mmol) containing molecular sieve (4 Å) in dichloromethane (5ml) was treated with dess-martin oxidant (1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1 ziodoxol) -3 (1)H) 623 mg, 1.47 mmol) and stirring at room temperatureAnd 5 min. The reaction mixture is then adsorbed onto kieselguhr and purified by column chromatography on silica gel (30-100% ethyl acetate/cyclohexane gradient) to yield 386 mg (72% of theory) of the title compound.

Intermediate 34A

(4-amino-7-bromopyrrolo [2,1-f ] [1,2,4] triazin-6-yl) methanol

A solution of 1, 3-dibromo-5, 5-dimethylhydantoin (87 mg, 0.305 mmol) in THF (1ml) was added dropwise to a solution of intermediate 8A (100 mg, 0.609 mmol) in THF (4ml) and methanol (2ml) at-78 ℃. The mixture was stirred at-78 ℃ for 16 h, then diluted with water and extracted with ethyl acetate. The combined organic layers were washed with saturated aqueous sodium chloride, dried over magnesium sulfate and evaporated. Purification by column chromatography on silica gel (dichloromethane/methanol 20:1 → 10:1) gives 55 mg (32% of theory) of the title compound.

Intermediate 35A

7-bromo-6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of intermediate 34A (885 mg, 3.64 mmol) in DMF (11 ml) was treated with tert-butyldimethylchlorosilane (823 mg, 5.46 mmol) and imidazole (743 mg, 10.92 mmol) and stirred at room temperature for 2 h. The reaction mixture was combined with 100 mg of the reaction mixture of the test run, diluted with water and extracted twice with ethyl acetate. The combined organic phases were washed with water and saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated. Purification by chromatography on silica gel (cyclohexane/ethyl acetate 2:1 → 100% ethyl acetate) gives 1.36 g (93% of theory) of the title compound.

Intermediate 36A

4-amino-6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

Intermediate 35A (880 mg, 2.46 mmol) and [1,1' -bis (diphenylphosphino) ferrocene were added under argon]Palladium (II) dichloride-dichloromethane complex [ PdCl₂(dppf) x DCM](120 mg, 0.148 mmol), bis (dibenzylideneacetone) Palladium [ Pd (dba)₂]A mixture of (135 mg, 0.148 mmol), zinc cyanide (578 mg, 4.92 mmol), zinc powder (64mg, 0.985 mmol) and zinc acetate (180 mg, 0.985 mmol) in degassed DMF/water (100:1, 9.2 ml) was stirred at 160 ℃ overnight. The reaction mixture was then combined with 100 mg of the reaction mixture of the test run and the combined mixture was adsorbed onto celite, filtered through another layer of celite and eluted with tert-butyl methyl ether. The filtrate was washed with saturated aqueous sodium bicarbonate solution, and the aqueous layer was extracted three more times with tert-butyl methyl ether. The combined organic phases were dried over sodium sulfate and evaporated. Purification by column chromatography on silica gel (cyclohexane/ethyl acetate 1:1) gives 453 mg (44% of theory) of the title compound.

Intermediate 37A

4-amino-5-bromo-6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of 1, 3-dibromo-5, 5-dimethylhydantoin (621 mg, 2.17 mmol) in THF (20ml) was added dropwise to a solution of intermediate 36A (1.1 g, 3.62 mmol) in THF (80 ml) at-50 ℃. The resulting mixture was slowly warmed to room temperature, stirred for 2h, and then quenched with 10% aqueous sodium thiosulfate and saturated aqueous sodium bicarbonate. The aqueous phase was extracted three times with ethyl acetate. The solid precipitated from the ethyl acetate solution was filtered off and washed with ethyl acetate to yield 508 mg (100% purity, 36% of theory) of the title compound as a first harvest. The remaining filtrate was dried over sodium sulfate and evaporated. The residue was precipitated from DMSO and washed with DMSO and ethyl acetate to yield further 498 mg (85% purity, 26% of theory) of the title compound.

Intermediate 38A

4-amino-6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of intermediate 37A (459 mg, 1.29 mmol) in degassed THF (14.8 ml) was added to (2' -aminobi-ethyl) under argonBenzol-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; 152mg, 0.19 mmol; see S.L. Buchwald et al, J. Am. Chem. Soc.132 (40) 14073-14075(2010)) and intermediate 6A (647 mg, 1.94 mmol). Degassed 0.5M aqueous potassium phosphate solution (5.1 ml) was added dropwise, and the resulting mixture was stirred at 60 ℃ for 2 h. The reaction mixture was then combined with the previously described reaction mixtures of 70 mg, 90 mg and 500mg test runs and evaporated. The residue is purified by column chromatography on silica gel (cyclohexane/ethyl acetate 3:1 → 100% ethyl acetate) to yield 1.0 g (58% of theory) of the title compound.

Intermediate 39A

4-amino-6-formyl-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of example 68 (250 mg, 0.684 mmol) in dichloromethane (5ml) was treated with dess-martin oxidant (1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1, 1-benziodoxol)H) 377 mg,0.889 mmol) and stirred at room temperature for 1 h. The reaction mixture was combined with the reaction mixture from 50 mg of the test run, quenched with saturated aqueous sodium bicarbonate and saturated aqueous sodium thiosulfate (1:1) and stirred at room temperature for 30 min. The aqueous phase was extracted three times with ethyl acetate and the combined organic phases were dried over sodium sulfate and evaporated. Purification by column chromatography on silica gel (cyclohexane/25% ethyl acetate → 100% ethyl acetate) gives 102 mg (24% of theory) of the title compound.

Intermediate 40A

4-amino-6- (azidomethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of example 68(100 mg, 273. mu. mol) in dichloromethane (5ml) was treated with thionyl chloride (39. mu.l, 547. mu. mol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in DMF (6 ml) and treated with sodium iodide (205 mg, 1.36 mmol) and sodium azide (355 mg, 5.47 mmol). The mixture was stirred at 80 ℃ overnight, then diluted with water and extracted three times with ethyl acetate. The combined organic layers were washed with water, then with saturated aqueous sodium chloride solution, dried over sodium sulfate and evaporated to give 91 mg of crude product, which was used in the next step without further purification.

Intermediate 41A

5, 7-dibromo-6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of intermediate 18A (2 g, 6.21 mmol) in DMF (20ml) was treated with imidazole (846 mg, 12.4mmol) and tert-butyldimethylsilyl chloride (1.12 g, 7.45 mmol) and stirred at room temperature for 20 h. The reaction mixture was then diluted with water (200 ml) and stirred at room temperature for a further 2 h. The solid is filtered off to yield 2.46 g (88% of theory) of the title compound.

Intermediate 42A

2- [ 4-amino-5-bromo-6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] propan-2-ol

A solution of intermediate 41A (1 g, 2.29mmol) in THF (40 ml) was cooled to-78 deg.C under argon and treated with methyllithium in 1.6M in ether (1.5 ml, 2.40 mmol). After stirring at-78 deg.C for 10 min, a 1.6M solution of n-butyllithium in hexane (1.58 ml, 2.52 mmol) was added and stirring continued for 10 min. Acetone (1.68 ml, 22.92 mmol) was added and the resulting mixture was slowly warmed to room temperature and stirred at room temperature for 18 h. The reaction was then quenched with water and the aqueous phase was extracted twice with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated. Purification by flash chromatography on silica gel (cyclohexane/ethyl acetate 2:1) gives 306 mg (30% of theory) of the title compound.

Intermediate 43A

2- [ 4-amino-5-bromo-6- (hydroxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] propan-2-ol

A solution of intermediate 42A (304 mg, 0.732 mmol) in THF (15 ml) was treated with tetra-n-butylammonium fluoride in THF1M solution (768. mu.l, 768. mu. mol) was treated and stirred at room temperature for 2 min. The reaction mixture was diluted with acetonitrile (20ml) then evaporated and the residue purified by preparative RP-HPLC (Reprosil C18, gradient 10-30% acetonitrile/0.2% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 180mg (67% of theory) of the title compound.

Intermediate 44A

4-amino-7- (2-hydroxypropan-2-yl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-6-carbaldehyde

A solution of example 73 (135 mg, 89% purity, 302 μmol) in dichloromethane (7 ml) was treated with Des-Martin oxidant (1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1, 1-benzidoxol)H) 166 mg, 392 μmol) and stirred at room temperature for 70 min. The reaction mixture was combined with 18mg (45 μmol) of the reaction mixture of the test run and quenched with saturated aqueous sodium bicarbonate and saturated aqueous sodium thiosulfate (1: 1). The aqueous phase was extracted three times with dichloromethane. The combined organic phases are washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to give 143 mg (77% purity, 92% of theory) of the title compound.

Intermediate 45A

6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of 1.5 g (9.14 mmol) of intermediate 8A in 15 ml of dry DMF was treated with 1.65 g (10.96mmol) of tert-butyldimethylchlorosilane and 1.24 g (18.27 mmol) of imidazole and stirred at room temperature overnight. The reaction mixture was poured into 250 ml of water and stirred for 5 min. The resulting precipitate was filtered off and dried in vacuo at 45 ℃. Yield: 2.28g (90% of theory).

Intermediate 46A

6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) -7-chloropyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of 2 g (7.18 mmol) of intermediate 45A in 20ml THF at-10 ℃ over 60 min was taken up with 893 mg (6.47 mmol)NChlorosuccinimide is treated in 6 parts. Stirring was continued at-10 ℃ for 15min, and then the mixture was warmed to room temperature. Another 192 mg portion (1.44 mmol) was added at room temperatureNChlorosuccinimide and stirring is continued overnight. About one tenth of the reaction mixture was evaporated to dryness and the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 94 mg (4% of theory) of the title compound. The major part of the reaction mixture is adsorbed onto silica gel and chromatographed on silica gel using isohexane/ethyl acetate 5-66% as eluent to give 899 mg (40% of theory) of the title compound. Total yield: 993 mg (44% of theory).

Intermediate 47A

5-bromo-6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) -7-chloropyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of 890 mg (2.85 mmol) of intermediate 46A in 20ml of DMF is taken up over 1h at-10 ℃ with 506 mg (2.85 mmol)NBromo-succinimide is treated in portions. Stirring was continued at-10 ℃ for 3h and then at room temperature overnight. Water (200 ml) was added and the mixture was stirred for 2 h. The precipitated solid was filtered off, washed with water and dried in vacuo at 45 ℃. Yield: 997 mg (89% of theory).

Intermediate 48A

6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) -7-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

800 mg (2.04 mmol) of intermediate 47A, 680 mg (2.04 mmol) of intermediate 6A, 80mg (0.1 mmol) of (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; see S.L. Buchwald et al, under an argon atmosphere, J. Am. Chem. Soc.132 (40) 14073-14075(2010)) and 1.3 g (6.13 mmol) of potassium phosphate were added to the flask. Then, the user can use the device to perform the operation,30 ml of degassed 1, 4-dioxane/water mixture (5:1) were added and the solution was stirred at 70 ℃ for 1 h. 680 mg (2.04 mmol) of intermediate 6A and 32mg (0.04 mmol) of (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1) are additionally added and stirring is continued at 70 ℃ for a further 1 h. This step was repeated three times until the starting material was consumed (controlled by LC-MS). With the last portion of reagent, 1.6 ml of 5M aqueous sodium hydroxide solution was also added to achieve a pH of 8-9. At the end of the reaction period, 30 ml of water and 5M aqueous formic acid (pH 3-4) were added, after which the oil was isolated. A portion of this oil was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). Two fractions of the title compound were obtained: 103 mg of solid (89% pure, 9% of theory, determined by LC-MS) and 23 mg of solid (100% pure, 2% of theory, determined by LC-MS). The remaining oil and supernatant were diluted with water, adjusted to pH 8-9 with saturated aqueous sodium bicarbonate and extracted three times with dichloromethane. The combined organic phases were washed with water, dried and evaporated under reduced pressure to give 2.1 g of an oil. This material was purified by column chromatography on silica gel using dichloromethane/0-5% methanol as eluent to give further 709 mg (43% purity by LC-MS, 31% of theory) of the title compound. Total yield: 42% of theory.

Intermediate 49A

4-amino-7-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-6-carbaldehyde

To a suspension of 166 mg (0.44 mmol) of example 79 and molecular sieve (3 Å) in 3ml of dichloromethane at 0-5 ℃ was added 207 mg (0.49 mmol) of dess-martin oxidant (1,1, 1-trisAcetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1H) -ketones). The mixture was stirred at this temperature for 10 min, then 56mg (0.13 mmol) of dess-martin oxidant was further added and stirring was continued at 5 ℃ for 15min and at room temperature for 10 min. Thereafter, the mixture was adsorbed onto celite and purified by silica gel column chromatography using dichloromethane/0-10% methanol as eluent. Yield: 100 mg of solid (92.7% purity, 56% of theory, determined by LC-MS).

Intermediate 50A

7-chloro-6- (chloromethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

The title compound was isolated as a by-product in preparation example 79 (see below). Yield: 9.2 mg (10% of theory).

Intermediate 51A

6- ({ [ tert-butyl (dimethyl) silyl ] oxy } methyl) -7-methylpyrrolo [2,1-f ] [1,2,4] triazin-4-amine

To a solution of 3g (8.4 mmol) of intermediate 35A in 60 ml of 1, 4-dioxane under an argon atmosphere was added 171 mg (0.21 mmol) of PdCl₂(dppf) xDCM, then stepwise over 10 min16.8 ml of a 2M solution of dimethylzinc in toluene (causing the temperature to rise from 22 ℃ to 31 ℃) are added dropwise. Stirring was first continued at room temperature for 10 min, then at 90 ℃ for 13 h. Thereafter, water (10 ml) was added to the reaction mixture at room temperature, and the suspension was stirred for 1 h. The mixture was evaporated under reduced pressure and the residue was dissolved in water and ethyl acetate and stirred for a further 1 h. The precipitate was filtered and discarded, the phases separated and the aqueous layer extracted twice with ethyl acetate. The combined organic phases were dried and evaporated, yielding 2.45 g (92% purity by LC-MS, 92% of theory) of the title compound.

Intermediate 52A

(4-amino-7-methylpyrrolo [2,1-f ] [1,2,4] triazin-6-yl) methanol

。

The method comprises the following steps:

to a solution of 100 mg (0.28 mmol) of intermediate 35A in 2ml of 1, 4-dioxane under an argon atmosphere was added 6mg (0.01 mmol) of PdCl₂(dppf) x DCM, then 0.56ml of a 2M solution of dimethylzinc in toluene was added dropwise over 10 min. The mixture was stirred at 90 ℃ overnight, then evaporated and the residue treated with acetonitrile and 5M aqueous formic acid. The precipitate was filtered off, the filtrate was evaporated, and the residue was dissolved in DMSO/acetonitrile and purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). Yield: 33 mg (66% of theory).

Method of producing a composite material2:

750 mg (3.1 mmol) of intermediate 34A, 515 μ l (3.7 mmol) of trimethylcyclotriboroxane, 786 mg (3.7 mmol) of potassium phosphate and 73 mg (0.09 mmol) of (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; see S.L. Buchwald et al, under an argon atmosphere, J. Am. Chem. Soc.132 (40) 14073-14075(2010)) is added to the microwave reaction vessel. Then, 13ml of degassed 1, 4-dioxane/water mixture (5:1) were added, the vessel was sealed and the mixture was heated in a microwave (4 bar, 50watt) to 140 ℃ for 20 min. A further 50 mg of (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1) are added and the mixture is heated again to 140 ℃ in the microwave for 20 min. Another 515 μ l of trimethylcyclotriboroxane was added and the vessel was heated again to 140 ℃ for 20 min. The latter procedure was repeated twice more with each 30min heating period until LC-MS showed that only a small amount of starting material remained. The mixture was filtered through celite, washed with 1, 4-dioxane, and the combined filtrates were evaporated to dryness. The residue was combined with the previous 100 mg test run and purified by preparative RP-HPLC (XBridge C18, gradient 5-42% acetonitrile/0.05% aqueous ammonium hydroxide). Yield: 238 mg (38% of theory).

Intermediate 53A

(4-amino-5-bromo-7-methylpyrrolo [2,1-f ] [1,2,4] triazin-6-yl) methanol

A solution of 245 mg (1.38 mmol) of intermediate 52A in 8.8 ml DMF at-10 ℃ over 2h was taken 288 mg (1.62 mmol)NBromo-succinimide is treated in portions. Stirring at-10 deg.CFor 30min, then at room temperature for 2 h. The mixture was poured into 50 ml of water and extracted with ethyl acetate. The combined organic phases were dried and evaporated. The residue was flash chromatographed on silica gel with dichloromethane/0-15% methanol as eluent. Yield: 148 mg (42% of theory).

Intermediate 54A

4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7-methylpyrrolo [2,1-f ] [1,2,4] triazine-6-carbaldehyde

To a suspension of 175 mg (0.49 mmol) of example 75 and molecular sieve (3 Å) in 3.5 ml of dichloromethane at 0-5 deg.C was added 230 mg (0.54 mmol) of dess-martin oxidant (1,1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy) -3(1 ziodoxol) -3 (1)H) -ketones). The mixture was stirred at this temperature for 5min, followed by stirring at room temperature for 30 min. Thereafter, the mixture was adsorbed onto celite and purified by flash chromatography on silica gel with isohexane/10-100% ethyl acetate as eluent. Yield: 139 mg of solid (79% of theory).

Intermediate 55A

4- [ (4-amino-6-methylpyrrolo [2,1-f ] [1,2,4] triazin-7-yl) methyl ] piperazine-1-carboxylic acid tert-butyl ester

A solution of 6-methylpyrrolo [2,1-f ] [1,2,4] triazin-4-amine (500 mg, 3.3 mmol; preparation described in PCT International patent application WO 2007/056170) in acetic acid (8 ml) was treated with 37% aqueous formaldehyde (328. mu.l, 4.04 mmol) and piperazine-1-carboxylic acid tert-butyl ester (754 mg, 4.04 mmol). The mixture was stirred at 60 ℃ overnight. After evaporation, the residue was dissolved in ethyl acetate and washed with saturated aqueous sodium bicarbonate. The aqueous layer was extracted twice with ethyl acetate. The combined organic phases were dried over magnesium sulphate and evaporated to give 1.2 g of crude product, which was used in the next step without further purification.

Intermediate 56A

4- [ (4-amino-5-bromo-6-methylpyrrolo [2,1-f ] [1,2,4] triazin-7-yl) methyl ] piperazine-1-carboxylic acid tert-butyl ester

A solution of intermediate 55A (1.17 g, 3.37 mmol) in THF (20ml) was cooled to-60 deg.C and treated with 1, 3-dibromo-5, 5-dimethylhydantoin (5.78 mg, 2.02 mmol). The mixture was stirred at-60 ℃ to-20 ℃ for 4 h. Thereafter, the reaction mixture was quenched with 10% aqueous sodium thiosulfate solution. Most of the THF solvent was evaporated, after which a solid precipitated. Filtration and recrystallization from acetone gave 862 mg (59% of theory) of the title compound.

Intermediate 57A

4- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6-methylpyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazine-1-carboxylic acid tert-butyl ester

A solution of intermediate 56A (100 mg, 235. mu. mol) in degassed 1, 4-dioxane (3 ml) was treated with intermediate 6A (93 mg, 282. mu. mol), dicyclohexyl (2',6' -dimethoxybiphenyl-2-yl) phosphine (S-Phos; 9.6mg, 23. mu. mol) and palladium diacetate (2.6 mg, 11. mu. mol). Degassed 3M aqueous potassium phosphate solution (588. mu.l) was added and the resulting mixture was stirred at 60 ℃ for 1 h. Additional portions of intermediate 6A (78 mg, 235 μmol) were added and stirring was continued at 60 ℃ overnight. The reaction mixture was quenched with 2M aqueous sodium hydroxide solution and the aqueous phase was extracted with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated. Purification by column chromatography on silica gel (cyclohexane/ethyl acetate 3:2) gives 82 mg (62% of theory) of the title compound.

Intermediate 58A

4- [ (4-amino-6-chloropyrrolo [2,1-f ] [1,2,4] triazin-7-yl) methyl ] piperazine-1-carboxylic acid tert-butyl ester

Following the procedure of intermediate 55A, 4 g (23.7 mmol) of 6-chloropyrrolo [2,1-f ] [1,2,4] triazin-4-amine (preparation described in PCT International patent application WO 2007/064883) were reacted to give 11.2 g of the title compound as crude product which was used in the next step without further purification.

After 100 mg of preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) of the crude material obtained above, 67 mg of a sample of the corresponding formate salt, tert-butyl 4- [ (4-amino-6-chloropyrrolo [2,1-f ] [1,2,4] triazin-7-yl) methyl ] piperazine-1-carboxylate formate salt, was isolated.

Intermediate 59A

4- [ (4-amino-5-bromo-6-chloropyrrolo [2,1-f ] [1,2,4] triazin-7-yl) methyl ] piperazine-1-carboxylic acid tert-butyl ester

Following the procedure for intermediate 56A, 11 g (30 mmol) of intermediate 58A were reacted after flash chromatography on silica gel (dichloromethane/methanol 10:1) and subsequent preparative RP-HPLC (Daiso C18, gradient 40-65% acetonitrile/water) to give 1.17 g (9% of theory) of the title compound.

Intermediate 60A

4- { [ 4-amino-6-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazine-1-carboxylic acid tert-butyl ester

Under an argon atmosphere, 140 mg (0.31 mmol) of intermediate 59A, 102 mg (0.46 mmol)Intermediate 5A, 16 mg (0.02 mmol) of PdCl₂(dppf) XDCM and 122 mg (1.15 mmol) sodium carbonate were added to the flask. After addition of 3ml of degassed 1, 2-dimethoxyethane/water (3:1), the suspension is stirred at 60 ℃ for 2.5 h. Additional portions of intermediate 5A (50 mg, 0.23 mmol) and PdCl were added₂(dppf) x DCM (8 mg, 0.01 mmol) and stirring at 40 ℃ was continued for 2.5 h. The subsequent steps are repeated once more until the starting material is consumed. Then, the reaction mixture was partially evaporated under reduced pressure, water was added, and the mixture was extracted with dichloromethane. The combined organic phases were washed with water and saturated aqueous sodium chloride solution, dried and evaporated under reduced pressure. The residue (322 mg) was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to give 49mg of the title compound and Boc-deprotected derivative 6-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f][1,2,4]Mixtures of triazin-4-amines. The mixture was used as such in the next reaction step (see example 85).

Intermediate 61A

4- { [ 4-amino-6- (methoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

A solution of intermediate 10A (5 g, 28.1 mmol) and piperazin-2-one (3.09 g, 30.9 mmol) in acetic acid (85ml) was treated with 37% aqueous formaldehyde (3.15 ml, 42.1 mmol) and stirred at 60 ℃ for 16 h. The volatiles were evaporated under reduced pressure and the residue was dissolved in methanol and adsorbed onto celite. Purification by chromatography on a silica gel column (5-10% methanol in dichloromethane) gives 3.91 g (46% of theory) of the title compound.

Intermediate 62A

4- { [ 4-amino-5-bromo-6- (methoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

A solution of intermediate 61A (3.9 g, 13.4 mmol) in DMF (50 ml) was cooled to 0 ℃ and taken up withN-treatment of a solution of bromosuccinimide (2.63 g,14.8 mmol) in DMF (6 ml). The mixture was stirred at 0 ℃ for 1 h. The solvent was then evaporated and the residue was dissolved in methanol and adsorbed onto celite. Purification by chromatography on a silica gel column (5-10% methanol in dichloromethane) gives 1.99 g (39% of theory) of the title compound.

Intermediate 63A

7- (chloromethyl) -6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine hydrochloride

A suspension of example 86 (1 g, 2.51 mmol) in toluene (60 ml) was treated dropwise with thionyl chloride (1.83ml, 25.1 mmol) and the mixture was stirred at room temperature overnight. The volatiles were evaporated under reduced pressure. The residue is evaporated under reduced pressure three more times with toluene to give 0.85 g (74% of theory) of the title compound, which is used immediately in the next step without further purification.

Intermediate 64A

4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carboxylic acid

A suspension of intermediate 17A (5 g, 12.6 mmol) in THF/water (10:1, 220 ml) was treated with a 2M solution of 2-methyl-2-butene in THF (31.5 ml, 63.1 mmol) and with sodium dihydrogen phosphate (6.96 g, 50.4 mmol). The mixture was stirred at room temperature for 5 min. Sodium chlorite (4.56 g, 50.44 mmol) was then added and the resulting mixture was stirred at room temperature for 20 h. The suspension was filtered and the resulting solid was washed with water to give 4.24 g (74% of theory) of the title compound, which was used in the next step without further purification.

Intermediate 65A

5, 7-dimethoxy-1-benzothiophene

To a solution of 1-benzothiophene-5, 7-diol (1.16 g, 6.98 mmol) in acetone (20ml) under argon was added potassium carbonate (2.89 g, 20.9 mmol) and methyl iodide (912 μ l, 14.6 mmol). The resulting mixture was stirred at reflux for 18 h. After cooling to room temperature, the mixture was treated with a 7M solution of ammonia in methanol (10 ml) for 30min and then adsorbed onto silica gel. Purification by chromatography on silica gel (cyclohexane/ethyl acetate 40:1) gives 0.52 g (32% of theory) of the title compound.

Intermediate 66A

(5, 7-dimethoxy-1-benzothien-2-yl) boronic acid

A1.6M solution of n-butyllithium in hexane (1.84 ml, 2.95 mmol) was added dropwise to a solution of intermediate 65A (520 mg, 2.68 mmol) in dry THF (5ml) at-70 ℃ under an argon atmosphere. After 1h at-70 ℃, triisopropyl borate (742 μ l, 3.21 mmol) was added and the mixture was stirred for 16 h while slowly warming to room temperature. Dichloromethane and saturated aqueous ammonium chloride solution were added and the pH was adjusted to 6 by addition of 1M hydrochloric acid. The organic phase was separated and the aqueous phase was extracted with dichloromethane. The combined organic phases were dried over magnesium sulfate, filtered and evaporated. The residue obtained is purified by chromatography on a column of silica gel (elution first with dichloromethane/methanol 40:1, then with methanol and finally with 4M hydrochloric acid 10:1 in methanol/1, 4-dioxane) to yield 631 mg (71% purity, 71% of theory) of the title compound.

Intermediate 67A

4-amino-6- (chloromethyl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

Intermediate 67A was isolated as a byproduct of the synthesis of intermediate 11A after silica gel column chromatography (dichloromethane/acetone 8:2 → 7: 3).

Intermediate 68A

4-amino-6- [ (3-oxopiperazin-1-yl) methyl ] pyrrolo [2,1-f ] [1,2,4] triazine-7-carbaldehyde

A solution of intermediate 67A (11.03 g, 52.4 mmol) and 2-oxopiperazine (6.82 g, 68.1mmol) in 331 ml DMF was treated with DIPEA (13.7 ml, 78.6 mmol) and stirred at room temperature overnight. The precipitate is filtered off, washed with DMF and ether and then dried in vacuo to yield 11.64 g of the title compound (89% purity, 72% of theory).

Intermediate 69A

4- { [ 4-amino-7- (hydroxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

To a solution of intermediate 68A (10.17 g, 89% purity, 33.01 mmol) in 1M hydrochloric acid (370 ml) and methanol (370 ml) was added zinc powder (12.1 g, 185 mmol) at room temperature and the mixture was stirred at room temperature for 18 h. Silica gel (100 g) was added and the volatiles were evaporated under reduced pressure. The residue was suspended in methanol, the volatiles were evaporated again under reduced pressure, and the residue was dried in vacuo. The solid is subjected to silica gel column chromatography (gradient dichloromethane/methanol 7M ammonia 10:1 → 3:1) to yield 6.39 g of the title compound (60% of theory).

Intermediate 70A

4- { [ 4-amino-5-bromo-7- (hydroxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one trifluoroacetate

A suspension of intermediate 69A (1 g) in methanol/water (10:1, 33 ml) was treated with trifluoroacetic acid (0.56ml, 7.24 mmol) to give a clear solution. Dropwise addition at 0 deg.CN-a solution of bromosuccinimide (708 mg, 3.98mmol) in methanol (30 ml) and the mixture was stirred at 0 ℃ for 1 h. The precipitate formed is filtered off and dried in vacuo to yield 700 mg of the title compound (41% of theory).

Preparation example:

example 1

4- { [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

。

The method comprises the following steps:

a solution of intermediate 13A (3 g, 7.84 mmol) in methanol (87 ml) was treated with acetic acid (0.898 ml,15.68 mmol), 2-oxopiperazine (1.17 g, 11.76 mmol) and sodium triacetoxyborohydride (4.98 g,23.53 mmol). The mixture was stirred at room temperature for 4.5 h. Additional portions of 2-oxopiperazine (392 mg, 3.9mmol) and sodium triacetoxyborohydride (3.3 g, 15.68 mmol) were added and the resulting mixture was stirred at 60 ℃ overnight. After evaporation, the residue was dissolved in saturated aqueous sodium bicarbonate and extracted with ethyl acetate. The combined organic phases are dried over sodium sulfate and evaporated, and the residue is purified by column chromatography on silica gel (dichloromethane/methanol 40:1 → 10: 1). The product thus obtained is triturated in methanol and filtered off to yield 540 mg (14% of theory) of the title compound. The methanol mother liquor was evaporated and the residue was purified by double RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). The fractions containing pure product were combined and neutralized with saturated aqueous sodium bicarbonate. The acetonitrile solvent was evaporated and the aqueous phase was extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate and evaporated to give 395 mg (13% of theory) of the title compound as a second crop.

The method 2 comprises the following steps:

a solution of intermediate 21A (291 mg, 0.82 mmol) in acetic acid (2.9 ml) was treated with 37% aqueous formaldehyde (104. mu.l, 1.39mmol) and 2-oxopiperazine (139 mg,1.39 mmol). The mixture was stirred at 60 ℃ for 3h and then evaporated. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gives 184 mg (44% of theory) of the title compound.

Example 2

4- { [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one dihydrochloride

A solution of example 1(100 mg, 214. mu. mol) in1, 4-dioxane (2ml) was treated with a 4M solution of hydrochloric acid in1, 4-dioxane (2ml, 8 mmol). The solvent was evaporated, leaving 130 mg (quantitative) of the title compound.

Example 3

(3R) -3- ({ [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-7-yl radical]Methyl } amino) pyrrolidin-2-one dihydrochloride

A solution of intermediate 13A (520 mg, 1.36 mmol) in methanol (15 ml) and acetic acid (156. mu.l, 2.7mmol) was treated with: (R) -3-Aminopyrrolidin-2-one (503 mg, 5.0 mmol) and triacetoxyborohydride (1.06 g, 5.0 mmol). The mixture was stirred at room temperature overnight and then evaporated. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) and lyophilization of the product thus obtained from a 4M solution of hydrochloric acid in1, 4-dioxane gave 272 mg (36% of theory) of the title compound.

Example 4

(3R) -3- ({ [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-7-yl radical]Methyl } amino) pyrrolidin-2-one

A solution of intermediate 13A (2.0 g, 5.2 mmol) in methanol (58 ml) and acetic acid (0.6 ml) was treated with: (R) -3-aminopyrrolidin-2-one (785 mg, 7.8 mmol) and triacetoxyborohydride (3.32 g, 15.6 mmol). The mixture was stirred at room temperature overnight. After this time, the reaction mixture was diluted with saturated aqueous sodium bicarbonate and extracted three times with ethyl acetate. The combined organic phases were dried over sodium sulfate and evaporated. The residue is purified by double column chromatography on silica gel (dichloromethane/methanol 40:1 to 10:1) to yield 957 mg (37% of theory) of the title compound.

Example 5

4- { [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

。

The method comprises the following steps:

a solution of intermediate 17A (2 g, 5.05 mmol) in THF (100 ml) was treated with 2-oxopiperazine (1.01 g, 10.1 mmol), sodium triacetoxyborohydride (1.07 g, 5.04 mmol) and acetic acid (0.29 ml,5.04 mmol) at 0 deg.C. The resulting mixture was stirred at 0 ℃ for 30 min. Four additional portions of sodium triacetoxyborohydride (1.07 g, 5.04 mmol) and acetic acid (0.29 ml,5.04 mmol) were added every 30 minutes and the resulting mixture was stirred at 0 ℃ for an additional 30min, then at 35 ℃ for 25 min and finally at room temperature overnight. The reaction was quenched with 10% aqueous sodium chloride and extracted twice with ethyl acetate. The combined organic phases were evaporated. The residue is purified by column chromatography on silica gel (dichloromethane/methanol 95:5 → 90:10) to yield 360 mg (17% of theory) of the compound described in example 86 (see below) and 1.82 g of the title compound in separate fractions. The title product thus obtained was suspended in ethanol (20ml), refluxed for 2h, then cooled to 15 ℃. The solid is filtered off and washed with ethanol to yield 1.63 g (67% of theory) of the pure title compound.

The method 2 comprises the following steps:

a solution of example 13 (930 mg, 1.9 mmol) in dichloromethane (18 ml) was treated with thionyl chloride (210. mu.l, 2.8 mmol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in ethanol (18 ml) and treated with DIPEA (670. mu.l, 3.8 mmol). The mixture was stirred at 70 ℃ for 2h and then evaporated. The residue was purified by silica gel column chromatography (dichloromethane/methanol 98:2 → 90: 10). The product thus obtained was triturated in an acetonitrile/diethyl ether mixture and filtered. The filtrate was evaporated and the residue was repurified by column chromatography on silica gel (dichloromethane/methanol 98:2 → 90: 10). Again, the product thus obtained was triturated in acetonitrile/diethyl ether and filtered. This step is repeated once more. The three solid batches thus obtained are combined, triturated again in acetonitrile/diethyl ether and finally filtered off to yield 600 mg (62% of theory) of the title compound.

The method 3 comprises the following steps:

a solution of intermediate 22A (720 mg, 89% pure, 1.74 mmol) in acetic acid (10 ml) was stirred with piperazin-2-one (261 mg, 2.61 mmol) at 60 ℃. After 0, 3 and 12 h, 37% aqueous formaldehyde solution (260 μ l, 3.48 mmol) was added thereto in three portions, respectively, and the mixture was stirred at 60 ℃ for a total of 24 h. The volatiles were then removed under reduced pressure and the residue was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The organic phase was washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated. The residue was dissolved in a mixture of methanol and dichloromethane, adsorbed onto celite, dried in vacuo, and purified by flash chromatography on silica gel (gradient 0-6% methanol/dichloromethane). The product fractions were combined, evaporated and repurified by preparative RP-HPLC (Reprosil C18, gradient 30-50% acetonitrile/0.2% aqueous TFA). The product fractions were combined again, diluted with saturated aqueous sodium bicarbonate and extracted with ethyl acetate. The organic phase is washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to yield 281 mg (31% of theory) of the title compound.

Example 6

4- { [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one dihydrochloride

A solution of intermediate 17A (60 mg, 69% purity, 104 μmol) in methanol (6 ml) was treated with 2-oxopiperazine (22 mg, 209 μmol), sodium cyanoborohydride (33 mg, 522 mmol) and acetic acid (12 μ l, 209 μmol). The mixture was stirred at 60 ℃ for 16 h and then filtered. The filtrate and residue were each purified by preparative RP-HPLC (Reprosil C18, gradient 40-60% acetonitrile/0.2% aqueous TFA). The product fractions are combined, diluted with 1M hydrochloric acid (3 ml) and evaporated to dryness to yield 46mg (79% of theory) of the title compound.

Example 7

(3R) -3- ({ [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-7-yl radical]Methyl } amino) pyrrolidin-2-one dihydrochloride

A solution of intermediate 17A (60 mg, 69% purity, 104. mu. mol) in methanol (3 ml) was treated with (CR) -3-aminopyrrolidin-2-one (22 mg, 209 μmol), sodium cyanoborohydride (33 mg, 522 μmol) and acetic acid (12 μ l, 209 μmol). The mixture was stirred at 60 ℃ for 4 h and then filtered. The filtrate was purified by preparative RP-HPLC (Reprosil C18, gradient 40-60% acetonitrile/0.2% aqueous TFA). The product fractions are combined, diluted with 1M hydrochloric acid and evaporated to dryness to yield 46mg (79% of theory) of the title compound.

Example 8

(3R) -3- ({ [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-7-yl radical]Methyl } amino) pyrrolidin-2-one

A solution of intermediate 17A (226 mg, purity 75%, 428 μmol) in methanol (4ml) was treated with (CR) -3-aminopyrrolidin-2-one (85 mg, 855 μmol), sodium cyanoborohydride (134 mg, 2.14 mmol) and acetic acid (49 μ l, 855 μmol). The mixture was stirred at room temperature for 1.5 h. Thereafter, the mixture was directly separated by preparative RP-HPLC (Reprosil C18, gradient 40-60% acetonitrile/0.2% aqueous TFA). The product fractions were combined, diluted with saturated aqueous sodium bicarbonate and extracted with ethyl acetate. The combined organic phases are washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to dryness to yield 180mg (88% of theory) of the title compound.

Example 9

N ²- { [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazin-7-yl radical]Methyl glycinamide dihydrochloride

A solution of intermediate 17A (60 mg, 69% pure, 104 μmol) in methanol (3 ml) was treated with glycylamine hydrochloride (23 mg, 209 μmol), sodium cyanoborohydride (32 mg, 522 mmol) and acetic acid (12 μ l, 209 μmol). The mixture was stirred at 60 ℃ for 16 h. After filtration, the filtrate was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product fractions were combined, diluted with 1M hydrochloric acid, evaporated to dryness and combined with the residue from the filtration step. This material was repurified by double prep RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product fractions are combined again, diluted with 1M hydrochloric acid and evaporated to dryness to give 7.4 mg (13% of theory) of the title compound.

Example 10

6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (morpholin-4-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of intermediate 17A (55 mg, purity 73%, 101 μmol) in methanol (3 ml) was treated with morpholine (18 mg, 202 μmol), sodium cyanoborohydride (19mg, 303 mmol) and acetic acid (18 μ l, 304 μmol). The mixture was stirred at 60 ℃ for 18 h. Other amounts of morpholine (18 mg, 202 μmol), sodium cyanoborohydride (19mg, 303 mmol) and acetic acid (18 μ l, 304 μmol) were added and stirring continued at 60 ℃ for a further 3 h. The resulting mixture was diluted with THF to dissolve the precipitate and separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 32mg (68% of theory) of the title compound.

Example 11

1- (4- { [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-1-yl) ethanone dihydrochloride

A solution of intermediate 17A (130 mg, 0.328 mmol) in methanol (8 ml) was usedNAcetyl piperazine (63 mg, 0.492 mmol), sodium cyanoborohydride (103 mg, 1.63 mmol), and acetic acid (37. mu.l, 0.655 mmol). The mixture was stirred at 60 ℃ for 3 h. This was then combined with 30 mg of the reaction mixture of the test run, evaporated and purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). The product thus obtained was lyophilized from 1, 4-dioxane, then dissolved in ethyl acetate and washed with saturated aqueous sodium bicarbonate. The organic layer was dried over magnesium sulfate, evaporated and lyophilized again from 1, 4-dioxane. Repurification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) followed by double column chromatography on silica gel (dichloromethane/methanol 98:2 → 4:1) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 49mg (18% of theory) of the title compound.

Example 12

[ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methanol di (formate)

A solution of intermediate 23A (95 mg, 152. mu. mol) in a 4M solution of hydrochloric acid/1, 4-dioxane (3.7 ml) was stirred at room temperature for 2 h. After evaporation, the residue is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 44mg (62% of theory) of the title compound.

Example 13

4- { [ 4-amino-6- (hydroxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

A solution of intermediate 20A (1.34 g, 3.96 mmol) in acetic acid (13.9 ml) was treated with 37% aqueous formaldehyde (501. mu.l, 6.6 mmol) and 2-oxopiperazine (670 mg, 6.6 mmol). The mixture was stirred at 50 ℃ for 2h and then evaporated. Purification by column chromatography on silica gel (dichloromethane/methanol 98:2 → 90:10) gives 942 mg (49% of theory) of the title compound.

Example 14

7-{[(3S) -3-amino-3-methylpyrrolidin-1-yl]Methyl } -6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazine-4-amine trihydrochloride

A solution of intermediate 21A (100 mg, 0.28 mmol) in acetic acid (1ml) was treated with 37% aqueous formaldehyde (25. mu.l, 0.33 mmol) and [ ((0.33 mmol))3S) -3-methylpyrrolidin-3-yl]Tert-butyl carbamate (Yoshida et al,Chem. Pharm. Bull.1996, 44 (7), 1376; 1386; 67 mg, 0.33 mmol). The mixture was stirred at room temperature for 3h and then evaporated. The residue was diluted with saturated aqueous sodium bicarbonate and solid potassium carbonate was added until no more gas was produced. The aqueous phase was extracted three times with ethyl acetate. The combined organic phases were dried over sodium sulfate and evaporated. The residue was dissolved in 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. After evaporation, the residue was purified by double preparative RP-HPLC (first Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid; then Shield RP18, gradient 5-50% methanol +0.1% aqueous TFA/0.1% aqueous TFA). The product thus obtained is lyophilized from a 4M solution of hydrochloric acid in1, 4-dioxane to yield 14 mg (8% of theory) of the title compound.

Example 15

7-{[(3S) -3-amino-3-methylpyrrolidin-1-yl]Methyl } -6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazine-4-amines

A solution of intermediate 17A (100 mg, 271. mu. mol) in acetic acid (1ml) was treated with 37% aqueous formaldehyde (24. mu.l, 326. mu. mol) and [ ((24. mu.l, 326. mu. mol))3S) -3-methylpyrrolidin-3-yl]Tert-butyl carbamate (Yoshida et al,Chem. Pharm. Bull.1996, 44 (7), 1376-1386; 65 mg, 326mu mol) treatment. The mixture was stirred at 60 ℃ for 4 h. Then, other amounts of 37% aqueous formaldehyde solution (10. mu.l, 136. mu. mol) and [ (R) ()3S) -3-methylpyrrolidin-3-yl]Tert-butyl carbamate (27mg, 136. mu. mol) and stirring was continued at 60 ℃ overnight. After evaporation, the residue was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. Solid potassium carbonate was added until no more gas was produced. The aqueous phase was extracted three times with ethyl acetate. The combined organic phases were dried over sodium sulfate, evaporated and purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). The product thus obtained was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 1 h. After evaporation, the residue is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 24mg (18% of theory) of the title compound.

Example 16

1- (4- { [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-1-yl) ethanone dihydrochloride

A solution of intermediate 21A (50 mg, 141. mu. mol) in acetic acid (1ml) was treated with 37% aqueous formaldehyde (4.6. mu.l, 169. mu. mol) andNtreatment with acetyl piperazine (21.6 mg, 169. mu. mol). The mixture was stirred at 75 ℃ for 3 h. After evaporation, the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). Freeze-drying from a 4M solution of hydrochloric acid in1, 4-dioxane gives 34mg (39% of theory) of the title compound.

Example 17

6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-aminecarboxylic acid salt

A solution of intermediate 23A (110 mg, 204. mu. mol) in dichloromethane (2.2 ml) was treated with thionyl chloride (29. mu.l, 408. mu. mol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in methanol (2.2 ml) and treated with DIPEA (39. mu.l, 224. mu. mol). The mixture was stirred at 70 ℃ for 1h and then evaporated. The residue was dissolved in 4M solution of hydrochloric acid in1, 4-dioxane (2.2 ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 44.9 mg (46% of theory) of the title compound.

Example 18

6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of intermediate 23A (80 mg, 317. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (22. mu.l, 297. mu. mol) and stirred at room temperature for 15 min. After evaporation, the residue was dissolved in ethanol (2ml) and treated with DIPEA (28 μ l, 163 μmol). The mixture was stirred at 70 ℃ for 1h and then evaporated. The residue was dissolved in 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 1 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 35 mg (50% of theory) of the title compound.

Example 19

6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine dihydrochloride

A solution of example 18 (50 mg, 107. mu. mol) in a 4M solution of hydrochloric acid/1, 4-dioxane (1ml) was stirred at room temperature for 15 min. After evaporation, 55 mg (93% of theory) of the title compound are obtained.

Example 20

1- (4- { [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-1-yl) ethanone

A solution of example 18 (70 mg, 150. mu. mol) in dichloromethane (2ml) and THF (0.8 ml) was treated with acetyl chloride (21. mu.l, 300. mu. mol) and sodium carbonate (127 mg, 1.2 mmol). The mixture was stirred at room temperature overnight. After evaporation, the residue is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 27mg (31% of theory) of the title compound.

Example 21

4- ({ 4-amino-6- [ (2-hydroxyethoxy) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl } methyl) piperazin-2-one formate

A solution of example 13 (60 mg, 132. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (14. mu.l, 198. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in ethylene glycol (500. mu.l) and stirred at 100 ℃ for 90 min. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gives 34mg (47% of theory) of the title compound.

Example 22

2- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methoxy } ethanol dihydrochloride

A solution of intermediate 23A (100 mg, 185. mu. mol) in dichloromethane (4ml) was treated with thionyl chloride (27. mu.l, 371. mu. mol). The mixture was stirred at room temperature for 20min and then evaporated. The residue was dissolved in ethylene glycol/THF (2:1, 1.5 ml) and stirred at 100 ℃ for 2 h. After evaporation, the residue is purified by preparative RP-HPLC (reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) followed by column chromatography on silica gel (dichloromethane/methanol +0.1% aqueous ammonia 98:2 → 90: 10). Freeze-drying from a 4M solution of hydrochloric acid in1, 4-dioxane gave 67 mg (64% of theory) of the title compound.

Example 23

6- (butoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-aminecarboxylic acid salt

A solution of intermediate 23A (100 mg, 185. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (27. mu.l, 371. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in 1-butanol (2ml) and heated to 70 ℃ for 1 h. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 22 mg (24% of theory) of the title compound.

Example 24

5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) -6- (propoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-aminedi (formate)

A solution of intermediate 23A (100 mg, 185. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (27. mu.l, 371. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in 1-propanol (2ml), treated with DIPEA (48. mu.l, 278. mu. mol) and stirred at room temperature for 1 h. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 15mg (16% of theory) of the title compound.

Example 25

6- [ (cyclopropylmethoxy) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine bis (formate)

A solution of intermediate 23A (100 mg, 185. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (27. mu.l, 371. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in cyclopropylmethanol (2ml), treated with DIPEA (48. mu.l, 278. mu. mol) and stirred at 70 ℃ for 2 h. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 29mg (30% of theory) of the title compound.

Example 26

6- [ (cyclobutyloxy) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

A solution of intermediate 23A (85 mg, 157. mu. mol) in dichloromethane (1.7 ml) was treated with thionyl chloride (23. mu.l, 315. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in cyclobutanol (1.7 ml), treated with DIPEA (41. mu.l, 236. mu. mol) and stirred at 70 ℃ for 2 h. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (1.7 ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 23 mg (28% of theory) of the title compound.

Example 27

6- (Isopropoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-aminecarboxylate

A solution of intermediate 23A (65 mg, 120. mu. mol) in dichloromethane (1.3 ml) was treated with thionyl chloride (17. mu.l, 241. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in 2-propanol (1.3 ml), treated with DIPEA (23. mu.l, 132. mu. mol) and stirred at 70 ℃ for 1 h. DIPEA (23. mu.l, 132. mu. mol) was added again and the mixture was stirred again for 1h at 70 ℃. Then, another portion of DIPEA (63. mu.l, 362. mu. mol) was added and stirring was continued at 90 ℃ for 3 h. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (1.3 ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 23 mg (34% of theory) of the title compound.

Example 28

6- [ (2-methoxyethoxy) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-aminecarboxylate

A solution of intermediate 23A (100 mg, 185. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (27. mu.l, 371. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in 2-methoxyethanol (2ml), treated with DIPEA (35. mu.l, 204. mu. mol) and stirred at 70 ℃ for 1 h. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 50 mg (50% of theory) of the title compound.

Example 29

5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) -6- [ (2,2, 2-trifluoroethoxy) methyl ] pyrrolo [2,1-f ] [1,2,4] triazin-4-aminecarboxylic acid salt

A solution of intermediate 23A (100 mg, 185. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (27. mu.l, 371. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in 2,2, 2-trifluoroethanol (2ml), treated with DIPEA (35. mu.l, 204. mu. mol) and stirred at 70 ℃ for 1 h. DIPEA (35. mu.l, 204. mu. mol) was added and stirring continued at room temperature for 1 h. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 23 mg (23% of theory) of the title compound.

Example 30

6- [ (2-aminoethoxy) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

A solution of intermediate 23A (150 mg, 278. mu. mol) in dichloromethane (5ml) was treated with thionyl chloride (40. mu.l, 556. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in THF (0.5ml) and treated with tert-butyl (2-hydroxyethyl) carbamate (1ml) and DIPEA (242. mu.l, 1.39 mmol). The mixture was stirred at 100 ℃ overnight. After evaporation, the residue was dissolved in1, 4-dioxane (10 ml), treated with a 4M solution of hydrochloric acid in1, 4-dioxane (10 ml) and stirred at room temperature for 1 h. Volatiles were removed under reduced pressure and the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). Further purification by preparative RP-HPLC (Shield RP18, 25% acetonitrile/75% 0.01% aqueous TFA) followed by lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 10 mg (6% of theory) of the title compound.

Example 31

{ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methoxy } acetic acid methyl ester

A solution of intermediate 23A (50 mg, 92. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (13. mu.l, 186. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in methyl glycolate (1ml), treated with DIPEA (80. mu.l, 464. mu. mol) and stirred at 70 ℃ for 2 h. After evaporation, the residue was dissolved in dichloromethane (1.6 ml), treated with trifluoroacetic acid (400 μ l, 5.19 mmol) and stirred at room temperature for 1 h. The mixture was then evaporated and the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). The fractions containing the title compound were neutralized with saturated aqueous sodium bicarbonate and evaporated. The residue was dissolved in water, and the mixture was extracted three times with dichloromethane. The combined organic phases are dried over sodium sulfate and evaporated to yield 21mg (43% of theory) of the title compound.

Example 32

{ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methoxy } acetic acid

A solution of intermediate 24A (200 mg, 327. mu. mol) in THF (14 ml) was treated with 2.5M aqueous lithium hydroxide (16 ml) and stirred at 80 ℃ for 2 h. The mixture was then combined with 27mg of the reaction mixture of the test run. The aqueous phase was extracted twice with THF and the combined organic phases were evaporated. The residue was dissolved in 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 3 h. After evaporation, the residue was purified by preparative RP-HPLC (reprosil c18, gradient 10-95% acetonitrile/0.1% aqueous TFA). Lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane and repurification by preparative RP-HPLC (Xbridge C18, gradient 5-95% acetonitrile/0.1% aqueous ammonium hydroxide) gave 13mg (7% of theory) of the title compound.

Example 33

2- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methoxy } acetamide

Intermediate 24A (200 mg, 327 μmol) was treated with a 7M solution of ammonia in methanol (10 ml) and stirred at room temperature overnight. The mixture was then combined with 20 mg of the reaction mixture of the test run and the solvent was evaporated. The residue was treated with a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. Evaporation and successive purification by preparative RP-HPLC RP-HPLC (first Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA; then Xbridge C18, gradient 5-95% acetonitrile/0.1% aqueous ammonium hydroxide) gave 5.5 mg (3% of theory) of the title compound.

Example 34

2- ({7- [ (4-acetylpiperazin-1-yl) methyl ] -4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl } methoxy) acetamide

A solution of example 33 (105 mg, 68% purity, 144. mu. mol) in THF/dichloromethane (1:2.5, 3.9 ml) was treated with sodium carbonate (179 mg, 1.6 mmol) and stirred at room temperature for 30 min. Acetyl chloride (30 μ l,424 μmol) was added and the resulting mixture was stirred at room temperature for 30min, then quenched with methanol (2ml) and evaporated. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gives 30 mg (85% purity, 34% of theory) of the title compound.

Example 35

5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6- (phenoxymethyl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-aminedi (formate)

A solution of intermediate 23A (100 mg, 185. mu. mol) in dichloromethane (2ml) was treated with thionyl chloride (27. mu.l, 371. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in THF (2ml), treated with phenol (174 mg, 1.85 mmol) and DIPEA (48. mu.l, 278. mu. mol) and stirred at 70 ℃ for 2 h. Additional amounts of phenol (174 mg, 1.85 mmol) and DIPEA (64. mu.l, 371. mu. mol) were added and stirring was continued overnight at 70 ℃. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 2 h. Evaporation and purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gave 8mg (8% of theory) of the title compound.

Example 36

5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6- [ (methylamino) methyl ] -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

Intermediate 25A (150 mg, 279. mu. mol) in THF (3 ml) was treated with acetic acid (32. mu.l, 559. mu. mol), a 2M solution of methylamine in THF (698. mu.l, 1.39mmol), and sodium triacetoxyborohydride (296 mg,1.39 mmol). The mixture was stirred at 60 ℃ for 2h and then evaporated. The residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (1.87 ml) and stirred at room temperature for 2 h. After evaporation, the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). Freeze-drying from a 4M solution of hydrochloric acid in1, 4-dioxane gives 79 mg (49% of theory) of the title compound.

Example 37

6- [ (dimethylamino) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

A solution of intermediate 25A (150 mg, 279. mu. mol) in THF (3 ml) was treated with acetic acid (32. mu.l, 559. mu. mol), a 2M solution of dimethylamine in THF (698. mu.l, 1.39mmol) and sodium triacetoxyborohydride (296 mg,1.39 mmol). The mixture was stirred at 60 ℃ for 2h and then evaporated. The residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (1.88 ml) and stirred at room temperature for 2 h. After evaporation, the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). Freeze-drying from a 4M solution of hydrochloric acid in1, 4-dioxane gives 83 mg (50% of theory) of the title compound.

Example 38

6- [ (ethylamino) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

A solution of intermediate 25A (60 mg, 111.8. mu. mol) in THF (1ml) was treated with a 2M solution of ethylamine in THF (83. mu.l, 167. mu. mol), sodium triacetoxyborohydride (118 mg, 559. mu. mol) and acetic acid (83. mu.l, 167. mu. mol). The mixture was stirred at 60 ℃ for 90 min. Additional amounts of 2M ethylamine solution (83. mu.l, 167. mu. mol) and sodium triacetoxyborohydride (71 mg, 335. mu. mol) were added and stirring was continued for a further 2h at 60 ℃. After evaporation, the residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (3 ml) and stirred at room temperature overnight. The mixture was evaporated and the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) and lyophilized from a 4M solution of hydrochloric acid in1, 4-dioxane. Repurification by preparative RP-HPLC (SunfireC18, 20% acetonitrile/80% 0.02% aqueous TFA) and lyophilization again from a 4M solution of hydrochloric acid in1, 4-dioxane gave 19mg (29% of theory) of the title compound.

Example 39

2- ({ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } amino) ethanolic trihydrochloride

A solution of intermediate 25A (150 mg, 279. mu. mol) in THF (3 ml) was treated with 2-aminoethanol (84. mu.l, 1.39mmol), sodium triacetoxyborohydride (296 mg,1.39 mmol) and acetic acid (32. mu.l, 559. mu. mol). The mixture was stirred at 60 ℃ for 2h and then evaporated. The residue was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (1.87 ml) and stirred at room temperature for 2 h. After evaporation, the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). Freeze-drying from a 4M solution of hydrochloric acid in1, 4-dioxane gives 140 mg (80% of theory) of the title compound.

General procedure for reductive amination reaction with intermediate 25A (GP 1):

a 0.1M solution of intermediate 25A in THF was treated with 5 equivalents of the corresponding amine component, 5 equivalents of sodium triacetoxyborohydride and 2 equivalents of acetic acid. The resulting mixture was stirred at 60 ℃ for 2h and then evaporated. A0.15M solution of the residue thus obtained in a 4M solution of hydrochloric acid/1, 4-dioxane was stirred at room temperature for 1-2 h. After evaporation, the residue was purified as described below.

Example 40

Rac-1- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperidin-3-ol trihydrochloride

Intermediate 25A (150 mg, 279. mu. mol) was reacted with 3-hydroxypiperidine (141 mg,1.39 mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 178 mg (quantitative) of the title compound.

EXAMPLE 41

1- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperidin-4-ol trihydrochloride

Intermediate 25A (150 mg, 279. mu. mol) was reacted with 4-hydroxypiperidine (141 mg,1.39 mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 162 mg (91% of theory) of the title compound.

Example 42

Rac-1- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } pyrrolidin-3-ol trihydrochloride

Intermediate 25A (150 mg, 279. mu. mol) was reacted with 3-hydroxypyrrolidine (113. mu.l, 1.39mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 148 mg (85% of theory) of the title compound.

Example 43

6- [ (diethylamino) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

Intermediate 25A (150 mg, 279. mu. mol) was reacted with diethylamine (144. mu.l, 1.39mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 127 mg (69% of theory) of the title compound.

Example 44

6- [ (cyclobutylamino) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

Intermediate 25A (150 mg, 279. mu. mol) was reacted with cyclobutyl amine (119. mu.l, 1.39mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 127 mg (72% of theory) of the title compound.

Example 45

5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) -6- (pyrrolidin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

Intermediate 25A (150 mg, 279. mu. mol) was reacted with pyrrolidine (116. mu.l, 1.39mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 112mg (64% of theory) of the title compound.

Example 46

6- [ (cyclopropylamino) methyl ] -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

Intermediate 25A (200 mg, 372. mu. mol) was reacted with cyclopropylamine (129. mu.l, 1.86mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 140 mg (62% of theory) of the title compound.

Example 47

6- { [ (cyclopropylmethyl) amino ] methyl } -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

Intermediate 25A (200 mg, 372. mu. mol) was reacted with cyclopropylmethylamine (161. mu.l, 1.86mmol) according to GP 1. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 163 mg (69% of theory) of the title compound.

Example 48

N- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f][1,2,4]Triazin-6-yl]Methyl glycine trihydrochloride

A solution of intermediate 25A (161 mg, 300. mu. mol) in THF (3.2 ml) was treated with 2-aminoacetic acid (112mg, 1.5 mmol), sodium triacetoxyborohydride (317 mg, 1.5 mmol) and acetic acid (34. mu.l, 600. mu. mol). The resulting mixture was stirred at 60 ℃ for 2h and then evaporated. The residue was purified by preparative RP-HPLC (reprosil c18, gradient 10-95% acetonitrile/0.1% aqueous TFA). The product thus obtained was dissolved in a 4M solution of hydrochloric acid in1, 4-dioxane (2ml) and stirred at room temperature for 1 h. After evaporation, the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). Subsequent lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane gave 18mg (9% of theory) of the title compound.

Example 49

4- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one trihydrochloride

A solution of intermediate 27A (220 mg, 354. mu. mol) in a 4M solution of hydrochloric acid/1, 4-dioxane (2ml) was stirred at room temperature for 2 h. The mixture was then evaporated, leaving 235 mg of crude product, which was used in the next step without further purification.

Example 50

[ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (morpholin-4-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methanol

A solution of intermediate 20A (500 mg, 87% pure, 1.28 mmol) and 4-methylen-morpholin-4-ium chloride (347mg, 2.56 mmol) in DMF (28 ml) was stirred at 70 ℃ for 1.5 h. The mixture was diluted with ethyl acetate and washed with saturated aqueous sodium bicarbonate and saturated aqueous sodium chloride. The organic layer is dried over magnesium sulfate and evaporated under reduced pressure to yield 710 mg (purity 78%, 99% of theory) of the title compound.

Example 51

(3S) -3- ({ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (morpholin-4-ylmethyl) pyrrolo [2,1-f][1,2,4]Triazin-6-yl]Methyl } amino) pyrrolidin-2-one

A solution of intermediate 33A (65 mg, 149. mu. mol) in methanol (2ml) was treated with (3S) -3-aminopyrrolidin-2-one (45mg, 446 μmol), sodium cyanoborohydride (47 mg, 743 μmol) and acetic acid (26 μ l, 446 μmol). After stirring for 16 h at 60 ℃ the resulting mixture was passed through a preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% TFA in waterLiquid) separation. The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 49mg (63% of theory) of the title compound.

Example 52

4- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (morpholin-4-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

A solution of intermediate 33A (65 mg, 149 μmol) in methanol (2ml) was treated with 2-oxopiperazine (45mg, 446 μmol), sodium cyanoborohydride (47 mg, 743 μmol) and acetic acid (26 μ l, 446 μmol). After stirring at 60 ℃ for 16 h, the resulting mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 41mg (53% of theory) of the title compound.

Example 53

Rac-1- ({ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (morpholin-4-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } amino) propan-2-ol

A solution of intermediate 33A (64mg, 146. mu. mol) in methanol (2ml) was racemized-1-amino propan-2-ol (33 mg, 439 μmol), sodium cyanoborohydride (46 mg, 731 μmol) and acetic acid (25 μ l, 439 μmol). After stirring at 60 ℃ for 16 h, the resulting mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 42mg (57% of theory) of the title compound.

Example 54

1- ({ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (morpholin-4-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } amino) -2-methylpropan-2-ol

A solution of intermediate 33A (80 mg, 183 μmol) in methanol (2ml) was treated with 1-amino-2-methylpropan-2-ol (34 mg, 274 μmol), sodium cyanoborohydride (57 mg, 914 μmol) and acetic acid (21 μ l, 366 μmol). After stirring at 60 ℃ for 16 h, the resulting mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratosphere)es SPE, PL-HCO₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 29mg (31% of theory) of the title compound.

Example 55

1- (4- { [ 4-amino-6- (hydroxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-1-yl) ethanone

A solution of intermediate 20A (345 mg, 1.01 mmol) in acetic acid (5ml) was treated with 37% aqueous formaldehyde (91. mu.l, 1.22 mmol) and 1-acetylpiperazine (160 mg, 1.22 mmol). The mixture was stirred at 60 ℃ for 6h and then evaporated. The residue was dissolved in a mixture of THF/1M aqueous lithium hydroxide solution (1:1, 10 ml) and stirred at room temperature for 2 h. The mixture was then combined with 100 mg of the reaction mixture of the test run and saturated aqueous ammonium chloride solution was added. The mixture is extracted with ethyl acetate and the organic phase is washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure to give 678 mg (87% purity, 94% of theory) of the title compound.

Example 56

(3R) -3- [ ({7- [ (4-acetylpiperazin-1-yl) methyl)]-4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazin-6-yl } methyl) amino]Pyrrolidin-2-ones

A solution of intermediate 31A (80 mg, 167. mu. mol) in methanol (1.4 ml) was treated with (C3R) -3-aminopyrrolidin-2-one (21 mg, 251 μmol), sodium cyanoborohydride (52 mg, 836 μmol) and acetic acid (19 μ l, 334 μmol). After stirring at 60 ℃ for 16 h, the resulting mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 46mg (49% of theory) of the title compound.

Example 57

1- (4- { [ 4-amino-6- { [ (2-hydroxy-2-methylpropyl) amino ] methyl } -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-1-yl) ethanone

A solution of intermediate 31A (80 mg, 183. mu. mol) in methanol (2ml) was treated with 1-amino-2-methylpropan-2-ol (31 mg, 251. mu. mol), sodium cyanoborohydride (53 mg, 836. mu. mol) and acetic acid (19. mu. l, 334. mu. mol). After stirring at 60 ℃ for 16 h, the resulting mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product thus obtained is dissolved in methanol and passed through an anionExchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 29mg (31% of theory) of the title compound.

Example 58

4- ({ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6- [ (3-oxopiperazin-1-yl) methyl ] pyrrolo [2,1-f ] [1,2,4] triazin-7-yl } methyl) piperazine-1-carbaldehyde formate salt

Acetic anhydride (498. mu.l, 5.17 mmol) and formic acid (237. mu.l, 6.28 mmol) were first stirred at 50 ℃ for 2h and then at room temperature overnight. Subsequently, the mixture was diluted with dichloromethane (5.1 ml) and 1.16 ml of this solution was added to a solution of example 49 (233 mg, 370 μmol) in pyridine (89 μ l). After stirring at room temperature for 2h, the mixture was diluted with methanol and then evaporated. The residue is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 77 mg (35% of theory) of the title compound.

Example 59

4- ({7- [ (4-acetylpiperazin-1-yl) methyl ] -4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl } methyl) piperazin-2-one

A suspension of example 49 (310 mg, 522. mu. mol) in dichloromethane/THF (2.5:1, 9.64 ml) was treated with sodium carbonate (442 mg, 4.17 mmol) and stirred at room temperature for 30 min. Acetyl chloride (74. mu.l, 1.04mmmol) was added and the resulting mixture was stirred at room temperature for 2 h. After quenching with methanol, the mixture is evaporated and the residue is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 133 mg (45% of theory) of the title compound.

Example 60

4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylcarbonyl) pyrrolo [2,1-f ] [1,2,4] triazine-6-carboxylic acid methyl ester bis (formate)

To a solution of intermediate 28A (15 mg, 26. mu. mol) in THF/methanol mixture (5:1, 180. mu.l) cooled to 0 deg.C was added (trimethylsilyl) diazomethane (2M solution in hexane, 15. mu.l, 32. mu. mol). The resulting mixture was slowly warmed to room temperature over 30min and then evaporated. The residue was dissolved in 4M solution of hydrochloric acid in1, 4-dioxane (0.5ml) and stirred at room temperature for 1 h. After evaporation, the residue is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 4.8 mg (35% of theory) of the title compound.

Example 61

5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6- (1, 3-oxazol-5-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

A solution of intermediate 25A (100 mg, 0.19 mmol) in methanol (3.33 ml) was treated with (4-toluenesulfonyl) methylisonitrile (36 mg, 0.19 mmol) and potassium carbonate (25 mg, 186. mu. mol). The mixture was refluxed for 6 h. It was then combined with the reaction mixture from 3 30 mg test runs and evaporated. The residue was dissolved in 4M solution of hydrochloric acid in1, 4-dioxane (10 ml) and stirred at room temperature for 2 h. After evaporation, the residue was purified by double prep RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). Further purification by column chromatography on silica gel (dichloromethane/methanol 5:1, containing 0.5% ammonia) and lyophilization from a 4M solution of hydrochloric acid in1, 4-dioxane yield 75 mg (34% of theory) of the title compound.

Example 62

6- (aminomethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

A suspension of intermediate 29A (76 mg, 134. mu. mol) and 10% Pd/C (60 mg) in a 0.5M solution of hydrochloric acid/methanol (20ml) under 1 atm hydrogen was stirred at room temperature for 3 h. The mixture was then filtered through celite, the filtrate was evaporated, and the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). The product thus obtained is lyophilized from a 4M solution of hydrochloric acid in1, 4-dioxane to yield 34mg (46% of theory) of the title compound.

Example 63

N- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f][1,2,4]Triazin-6-yl]Methyl } acetamide bis (trifluoroacetate)

A solution of intermediate 30A (210 mg, 362. mu. mol) in dichloromethane (26 ml) was treated with trifluoroacetic acid (5.2 ml) and stirred at room temperature for 1 h. After evaporation at room temperature, the residue is purified by preparative RP-HPLC (reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA) to yield 163 mg (63% of theory) of the title compound.

Example 64

N- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f][1,2,4]Triazin-6-yl]Methyl acetamide dihydrochloride

A solution of intermediate 30A (80 mg, 0.14 mmol) in dichloromethane (10 ml) and trifluoroacetic acid (2ml) was stirred at room temperature for 1h and then evaporated. Purification by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) and lyophilization from a mixture of methanol and a 4M solution of hydrochloric acid in1, 4-dioxane gave 39 mg (50% of theory) of the title compound.

Example 65

N- ({ 4-amino-7- [ (4-formylpiperazin-1-yl) methyl group]-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazin-6-yl methyl) acetamide formate salt

Acetic anhydride (304. mu.l, 3.16 mmol) and formic acid (145. mu.l, 3.16 mmol) were first stirred at 50 ℃ for 2h and then at room temperature overnight. Subsequently, the mixture was diluted with dichloromethane (3.1 ml) and 663 μ l of this solution was added to a solution of example 63 (160 mg, 226 μmol) in pyridine (54 μ l). The mixture was stirred at room temperature for 2h, then diluted with methanol (1ml) and stirred at 40 ℃ for a further 2 h. After evaporation, the residue is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 74 mg (61% of theory) of the title compound.

Example 66

N- ({7- [ (4-acetylpiperazin-1-yl) methyl)]-4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazin-6-yl methyl) acetamides

A solution of example 63 (110 mg, 229. mu. mol) and acetyl chloride (32. mu.l, 458. mu. mol) in THF/dichloromethane (1:2, 3 ml) was treated with sodium carbonate (194 mg, 1.83 mmol) and stirred at room temperature overnight. Then, the mixture was diluted with methanol (2ml) and water (1ml) and stirred at room temperature for 1 h. After evaporation, the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). Freeze-dried from 1, 4-dioxane and repurified by column chromatography on silica gel (dichloromethane/methanol 50:1 → 100% methanol) to yield 35 mg (28% of theory) of the title compound.

Example 67

N- ({ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- [ (3-oxopiperazin-1-yl) methyl)]Pyrrolo [2,1-f][1,2,4]Triazin-6-yl methyl) acetamides

A suspension of intermediate 32A (35 mg, 73. mu. mol) in methanol (4.1 ml) was treated with acetic anhydride (13. mu.l, 146. mu. mol) and 10% Pd/C (41 mg) under 1 atm hydrogen and stirred at room temperature for 1 h. Filtration through celite and evaporation of the filtrate yield 30 mg (79% of theory) of the title compound.

Example 68

4-amino-6- (hydroxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of intermediate 38A (1 g, 2.08 mmol) in THF (10 ml) was treated with a 1M solution of tetra-n-butylammonium fluoride in THF (12 ml, 12 mmol). The mixture was stirred at room temperature overnight and then evaporated. The residue was dissolved in water and extracted three times with ethyl acetate. The combined organic phases were dried over magnesium sulfate and evaporated. The residue is triturated in tert-butyl methyl ether and the solid is filtered off to yield 680 mg (78% of theory) of the title compound.

Example 69

4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of example 68 (60 mg, 164. mu. mol) in dichloromethane (5ml) was treated with thionyl chloride (18. mu.l, 246. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in methanol (2ml) and treated with DIPEA (57. mu.l, 328. mu. mol). The mixture was first stirred at 60 ℃ for 2h, then refluxed overnight and finally heated to 150 ℃ in a microwave apparatus for 30 min. Thereafter, the mixture was evaporated and the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 18mg (28% of theory) of the title compound.

Example 70

4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of example 68 (60 mg, 164. mu. mol) in dichloromethane (5ml) was treated with thionyl chloride (18. mu.l, 246. mu. mol). The mixture was stirred at room temperature for 15min and then evaporated. The residue was dissolved in ethanol (2ml) and treated with DIPEA (57. mu.l, 328. mu. mol). The mixture was stirred at 60 ℃ overnight and then heated to 150 ℃ in a microwave device for 30 min. Thereafter, the mixture was evaporated and the residue was purified by preparative RP-HPLC (reprosil c18, gradient 10-95% acetonitrile/0.1% aqueous TFA). The product thus obtained was dissolved in ethanol (2ml) and heated again to 150 ℃ in a microwave oven for 30 min. DIPEA (57. mu.l, 328. mu. mol) was added and heated to 150 ℃ in a microwave oven for another 30 min. After evaporation, the residue is purified by means of a column chromatography on silica gel (dichloromethane/methanol 95:5) to yield 16 mg (23% of theory) of the title compound.

Example 71

4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6- [ (3-oxopiperazin-1-yl) methyl ] pyrrolo [2,1-f ] [1,2,4] triazine-7-carbonitrile

A solution of intermediate 39A (20 mg, 55. mu. mol) in THF (0.73 ml) was treated with acetic acid (6. mu.l, 110. mu. mol), 2-oxopiperazine (27mg, 275. mu. mol) and sodium triacetoxyborohydride (58 mg, 275. mu. mol). After stirring at room temperature for 3h, additional amounts of THF (1ml), acetic acid (6. mu.l, 110. mu. mol), 2-oxopiperazine (27mg, 275. mu. mol) and sodium triacetoxyborohydride (58 mg, 275. mu. mol) were added and stirring continued at room temperature overnight. After evaporation, the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 13mg (52% of theory) of the title compound.

Example 72

N,N'- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f][1,2,4]Triazine-6, 7-diyl]Bis (methylene) } diacetamide

A suspension of intermediate 40A (crude, 85 mg), 10% Pd/C (115 mg) and acetic anhydride (40 μ l, 435 μmol) in methanol (12 ml) under 1 atm hydrogen was stirred at room temperature. After 4 h, other amounts of 10% Pd/C (115 mg) and acetic anhydride (40. mu.l, 435. mu. mol) were added and stirring continued at room temperature for 2h under 1 atm hydrogen. The resulting mixture was filtered through celite, the filtrate was evaporated, and the residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous TFA). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 18mg (18% of theory) of the title compound.

Example 73

2- [ 4-amino-6- (hydroxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] propan-2-ol

A flask containing a suspension of intermediate 43A (180 mg, 598 μmol), intermediate 6A (299 mg, 897 μmol) and cesium fluoride (454 mg, 2.99 mmol) in THF/water (10:1, 11 ml) was degassed under reduced pressure and then refilled with argon. Addition of 4- (di-tert-butylphosphino) -N,NDimethylaniline-dichloropalladium (2:1; 13mg, 18 μmol). The resulting mixture was degassed again and stirred under argon at 50 ℃ for 16 h. Thereafter, the reaction mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 30-50% acetonitrile/0.2% aqueous TFA). The product fractions were diluted with a 7M solution of ammonia in methanol and then concentrated under reduced pressure. The precipitate is filtered off, washed with water and dried in vacuo to yield 99 mg (42% of theory) of the title compound.

Example 74

4- { [ 4-amino-7- (2-hydroxypropan-2-yl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

A solution of intermediate 44A (50 mg, 73% purity, 92 μmol) in methanol (4ml) was treated with 2-oxopiperazine (28 mg, 276 μmol), sodium cyanoborohydride (23 mg, 368 μmol) and acetic acid (21 μ l, 368 μmol). The mixture was first stirred at 60 ℃ for 18 h and then at room temperature for 3 days. Thereafter, the mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous TFA). The product fractions were diluted with a 7M solution of ammonia in methanol and evaporated under reduced pressure. The residue was dissolved in ethyl acetate and washed with water. The organic phase was dried over sodium sulfate and concentrated under reduced pressure. The residue is dissolved in1, 4-dioxane and lyophilized to yield 24mg (52% of theory) of the title compound.

Example 75

[ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7-methylpyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methanol

165 mg (0.64 mmol) of intermediate 53A, 143 mg (0.64 mmol) of intermediate 5A, 25 mg (0.03 mmol) of (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; see S.L. Buchwald et al, under an argon atmosphere, J. Am. Chem. Soc.132 (40) 14073-14075(2010)) and 409 mg (1.93 mmol) of potassium phosphate were added to the flask. Then, 7 ml of a degassed mixture of 1, 4-dioxane and water (5:1) was added and the solution was stirred at 70 ℃ for 1 h. Another portion of intermediate 5A (142 mg,0.64 mmol) and (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (10 mg, 0.012 mmol) was added and stirring continued for 1 h. Thereafter, the reaction mixture was partially evaporated under reduced pressure, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were evaporated under reduced pressure and the residue triturated with acetonitrile. The precipitate is filtered off and dried in vacuo to yield 180mg (92% purity by LC-MS, 73% of theory) of the title compound. A second crop (41 mg, 18% of theory) was obtained from the filtrate by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). Total yield: 91% of theory.

Example 76

4- { [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7-methylpyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

A suspension of 55 mg (0.16 mmol) of intermediate 54A in THF (1.5 ml) was treated with 78 mg (0.78mmol) of piperazin-2-one, 18 μ l (0.31 mmol) of acetic acid and 166 mg (0.78mmol) of triacetoxyborohydride. The mixture was stirred at room temperature overnight. Then, 1.5 ml of water was added, and most of the THF solvent was evaporated under reduced pressure. The remaining mixture was diluted with more water and the precipitated solid was filtered off and dried (35 mg). This material was further purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid). The product fractions were adjusted to pH 9 with solid potassium carbonate and partially concentrated under reduced pressure. The precipitated solid is filtered off and dried in vacuo at 45 ℃ to yield 16 mg (24% of theory) of the title compound.

Example 77

1- ({ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7-methylpyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } amino) -2-methylpropan-2-ol formate salt

A suspension of 55 mg (0.16 mmol) of intermediate 54A in THF (1.5 ml) was treated with 98 mg (0.78mmol) of 1-amino-2-methylpropan-2-ol hydrochloride, 39 mg (0.47 mmol) of sodium acetate and 166 mg (0.78mmol) of sodium triacetoxyborohydride. The mixture was stirred at room temperature overnight. Then, 1.5 ml of water was added, and the mixture was evaporated under reduced pressure. The residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to give 38 mg (52% of theory) of the title compound.

Example 78

1- ({ [ 4-amino-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7-methylpyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } amino) -2-methylpropan-2-ol

A solution of 29mg (0.06 mmol) of example 77 in 3ml of methanol was run through Stratospheres SPE PL-HCO preconditioned with 2ml of methanol₃MP-resin column. The column is washed with 4ml of methanol and the eluate is evaporated to yield 21.5 mg (82% of theory) of the title compound.

Example 79

[ 4-amino-7-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methanol

A solution of intermediate 48A (117 mg, 0.24 mmol) in 5ml THF was treated with 5ml concentrated HCl and stirred at room temperature overnight. Then, 12 ml of 5M aqueous sodium hydroxide solution and ethyl acetate were added, the layers were separated, and the aqueous layer was extracted twice with ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, dried and evaporated. The residue was purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to give 30 mg (34% of theory) of the title compound.

9.2 mg (10% of theory) of 7-chloro-6- (chloromethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine are isolated as a by-product (reference intermediate 50A).

Example 80

4- { [ 4-amino-7-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

The title compound was prepared according to the procedure for example 76, starting from 40 mg (0.11 mmol) of intermediate 49A. Yield: 27mg (55% of theory).

Example 81

1- ({ [ 4-amino-7-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } amino) -2-methylpropan-2-ol formate salt

The title compound was prepared according to the procedure for example 77, starting from 40 mg (0.11 mmol) of intermediate 49A. Yield: 24mg (45% of theory).

Example 82

1- ({ [ 4-amino-7-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } amino) -2-methylpropan-2-ol

The title compound was prepared from example 81 according to the procedure for example 78.

Example 83

7-chloro-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine

8.2 mg (0.02 mmol) of intermediate 50A was suspended in 1ml ethanol, treated with 41 μ l (0.11 mmol) of a 2.68M solution of sodium ethoxide/ethanol and refluxed for 1 min. The clear solution is then evaporated and the crude product is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) to yield 5mg (56% of theory) of the title compound.

Example 84

5- (7-methoxy-5-methyl-1-benzothien-2-yl) -6-methyl-7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-aminecarboxylate

A solution of intermediate 57A (100 mg, 191. mu. mol) in a 4M solution of hydrochloric acid/1, 4-dioxane (2ml) was stirred at room temperature for 3h and then evaporated. Purification by means of double preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous formic acid) gives 71 mg (67% of theory) of the title compound.

Example 85

6-chloro-5- (7-methoxy-5-methyl-1-benzothien-2-yl) -7- (piperazin-1-ylmethyl) pyrrolo [2,1-f ] [1,2,4] triazin-4-amine trihydrochloride

Intermediate 60A (65 mg, 0.12 mmol) was stirred in 1ml of a 4M solution of hydrochloric acid/1, 4-dioxane at room temperature for 2 h. The suspension is evaporated to dryness and the crude product is purified by preparative RP-HPLC (Reprosil C18, gradient 10-95% acetonitrile/0.1% aqueous hydrochloric acid) to yield 49mg (74% of theory) of the title compound.

Example 86

[ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methanol

The title compound (360 mg) was obtained as a by-product in preparation example 5 by preparation method 1.

Example 87

1- { [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } imidazolidin-2-one

A suspension of intermediate 63A (100 mg, 0.221 mmol) in THF (5ml) was suspended with imidazolidin-2-one (57 mg, 0.662 mmol) andN,Ndiisopropylethylamine (153. mu.l, 0.926 mmol) and the mixture was heated to 150 ℃ in a microwave oven for 90 min. Thereafter, the reaction mixture was purified by preparative RP-HPLC (Reprosil C18, gradient 40-60% acetonitrile/0.2% aqueous trifluoroacetic acid). The product thus obtained was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column was eluted with methanol and the filtrate was evaporated. The product is purified once more by preparative thin layer chromatography on silica gel (cyclohexane/ethyl acetate 3:1) to yield 24mg (22% of theory) of the title compound.

Example 88

4- { [ 4-amino-5- (7-methoxy-1-benzothien-2-yl) -6- (methoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

To a solution of intermediate 62A (72.6 mg, 197. mu. mol), (7-methoxy-1-benzothien-2-yl) boronic acid (45mg, 216. mu. mol) and cesium fluoride (149 mg, 983. mu. mol) in degassed THF/water (10:1, 2.2 ml) under argon was added (2' -aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4', 6-Triisopropylbiphenyl-2-yl) phosphine (1:1; 7.7 mg, 9.8 μmol; see S.L. Buchwald et al, J. Am. Chem. Soc.132 (40),14073-14075 (2010)). The resulting mixture was degassed again and stirred at 60 ℃ for 3h under argon. Thereafter, the mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.1% aqueous TFA). The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratosphere SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 31 mg (35% of theory) of the title compound.

Example 89

4- { [ 4-amino-6- (methoxymethyl) -5- (5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

To a solution of intermediate 62A (50 mg, 135. mu. mol), (5-methyl-1-benzothien-2-yl) boronic acid (28.6 mg, 149. mu. mol) and cesium fluoride (103 mg, 677. mu. mol) in degassed THF/water (10:1, 4.4 ml) under argon was added (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; 5.3 mg, 6.8. mu. mol; see S.L. Buchwald et al, J. Am. Chem. Soc.132 (40),14073-14075 (2010)). The resulting mixture was degassed again and stirred under argon at 60 ℃ for 16 h. Thereafter, the mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 30-50% acetonitrile/0.1% aqueous TFA). The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratosphere SPE, PL-HCO)₃MP-treeFat) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 27mg (45% of theory) of the title compound.

Example 90

1- [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] ethanol

A 1M solution of methyl magnesium bromide in THF (630 μ l, 630 μmol) was added dropwise to a solution of intermediate 17A (100 mg, 252 μmol) in THF (10 ml) at room temperature under argon. The mixture was stirred at room temperature for 3h, then treated with another portion of methyl magnesium bromide/THF (177 μ l, 177 μmol). The reaction mixture was stirred for a further 16 h, then quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and evaporated. The residue was purified by preparative RP-HPLC (Reprosil C18, gradient 40-60% acetonitrile/0.1% aqueous TFA). The product fractions were diluted with saturated aqueous sodium bicarbonate and extracted with ethyl acetate. The organic phase is washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered and evaporated to yield 39 mg (36% of theory) of the title compound.

Example 91

[ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] (cyclopropyl) methanol

A 0.5M solution (1.26 ml, 630 μmol) of cyclopropylmagnesium bromide in THF was added dropwise to a solution of intermediate 17A (100 mg, 252 μmol) in THF (5ml) at room temperature under argon. The mixture was stirred at room temperature for 1h, then quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and evaporated. The residue was purified by preparative RP-HPLC (Reprosil C18, gradient 50-70% acetonitrile/0.1% aqueous TFA). The product fractions were diluted with saturated aqueous sodium bicarbonate and extracted with ethyl acetate. The organic phase is washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered and evaporated to yield 10 mg (10% of theory) of the title compound.

Example 92

(3S) -3- ({ [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-7-yl radical]Methyl } amino) pyrrolidin-2-one

A suspension of intermediate 13A (100 mg, 0.233 mmol) in THF (2ml) was treated with (S) Treatment with-3-aminopyrrolidin-2-one (35 mg, 0.349 mmol), sodium triacetoxyborohydride (148 mg, 0.698 mmol) and acetic acid (26.6 μ l, 0.465 mmol). Will be provided withThe resulting mixture was stirred at room temperature for 3h and purified directly by preparative RP-HPLC (Reprosil C18, gradient 40-60% acetonitrile/0.2% aqueous trifluoroacetic acid). The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 56mg (51% of theory) of the title compound.

Example 93

(3S) -3- ({ [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2, 1-f)][1,2,4]Triazin-7-yl radical]Methyl } amino) pyrrolidin-2-one

A suspension of intermediate 17A (100 mg, 0.252 mmol) in THF (2ml) was treated with: (S) -3-aminopyrrolidin-2-one (38 mg, 378 μmol), sodium triacetoxyborohydride (160 mg, 757 μmol) and acetic acid (30 μ l, 504 μmol). The resulting mixture was stirred at room temperature for 3h, then directly purified by preparative RP-HPLC (reprosil c18, gradient 30-50% acetonitrile/0.2% aqueous trifluoroacetic acid). The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 84 mg (69% of theory) of the title compound.

General procedure for the preparation of examples 94-105 in Table I:

a 0.13M suspension of intermediate 17A in THF was treated with 1.5 equivalents of the corresponding amine component, 3 equivalents of sodium triacetoxyborohydride and 1.5 equivalents of acetic acid. The resulting mixture was stirred at 60 ℃ for 3-20 h. Thereafter, purification was performed according to the indicated method.

Example 106

4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) -N-[(3R) -2-oxopyrrolidin-3-yl]Pyrrolo [2,1-f][1,2,4]Triazine-7-carboxamides

A stirred solution of intermediate 64A (50 mg, 121 μmol) in DMF (2ml) at room temperature was usedN-[(1H-benzotriazol-1-yloxy) (dimethylamino) methylene]-NMethyl ammonium tetrafluoroborate (TBTU) (43mg, 133 μmol) and DIPEA (53 μ l, 303 μmol). After 15min, add (3R) -3-aminopyrrolidin-2-one (24 mg,242 μmol) and the resulting mixture was stirred at room temperature for a further 2 h. Thereafter, the mixture was passed through preparative RP-HPLC (Reprosil C18, gradient 30-50% acetonitrile/0.2% trifluoroacetic acidAqueous solution) was separated. The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 36mg (60% of theory) of the title compound.

Example 107

4- { [ 4-amino-6- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] carbonyl } piperazin-2-one

A stirred solution of intermediate 64A (50 mg, 121 μmol) in DMF (2ml) was treated with TBTU (43mg, 133 μmol) and DIPEA (53 μ l, 303 μmol) at room temperature. After 15min, piperazin-2-one (24 mg,242 μmol) was added, and the resulting mixture was stirred at room temperature for a further 16 h. Thereafter, the mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 40-60% acetonitrile/0.2% aqueous trifluoroacetic acid). The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 45mg (68% of theory) of the title compound.

General procedure for the preparation of example 108-:

a 0.13M solution of intermediate 64A in DMF was treated with 1.1 eq TBTU and 2.5 eq DIPEA and stirred at rt for 15 min. 2 equivalents of the corresponding amine were added and the resulting mixture was stirred at room temperature for a further 18 h. Thereafter, purification was performed according to the indicated method.

Example 124

4- { [ 4-amino-5- (5, 7-dimethoxy-1-benzothien-2-yl) -6- (methoxymethyl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

To a solution of intermediate 62A (100 mg, 271. mu. mol), intermediate 66A (77 mg, 325. mu. mol) and cesium fluoride (206 mg, 1.35 mmol) in degassed THF/water (10:1; 5ml) under argon was added (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; 42.6 mg, 54. mu. mol; see S.L. Buchwald et al), J. Am. Chem. Soc.132 (40),14073-14075 (2010)). The resulting mixture was degassed again and stirred under argon at 60 ℃ for 6 h. Then, another portion of intermediate 66A (3) was added9mg, 162 μmol) and stirring was continued at 60 ℃ for 10 h. Thereafter, the reaction mixture was separated by preparative RP-HPLC (Reprosil C18, gradient 30-50% acetonitrile/0.1% aqueous TFA). The product fractions were combined and evaporated to dryness. The residue was dissolved in methanol and passed through an anion exchange column (Stratospheres SPE, PL-HCO)₃MP-resin) filtration. The column is eluted with methanol and the filtrate is evaporated, yielding 40 mg (28% of theory) of the title compound.

Example 125

4- { [ 4-amino-7- (hydroxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

To a suspension of intermediate 70A (700 mg, 1.49 mmol), intermediate 5A (497 mg, 2.24 mmol) and cesium fluoride (1.36 g, 8.95 mmol) in degassed THF/water (2:1, 90 ml) under argon was added (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; 117 mg, 0.149mmol; see S.L. Buchwald et al, supra, J. Am. Chem. Soc.132 (40), 14073-14075(2010)). The resulting mixture was degassed again and stirred under argon at 60 ℃ overnight. Another portion of intermediate 5A (231 mg, 1.04 mmol) and (2 '-aminobiphenyl-2-yl) (chloro) palladium-dicyclohexyl (2',4',6' -triisopropylbiphenyl-2-yl) phosphine (1:1; 117 mg, 0.149 mmol) were added and stirring continued at 60 ℃ for 3 h. The precipitate was filtered off, washed with THF and dried in vacuo. The solid was suspended in a mixture of DMF and 1M aqueous trifluoroacetic acid and filtered. The filtrate was separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous trifluoroacetic acid).The product fractions were combined and then basified by addition of saturated aqueous sodium bicarbonate. The solution was extracted with ethyl acetate and the organic layer was washed with brine, dried over magnesium sulfate and evaporated to yield 88 mg of the title compound (13% of theory).

Example 126

4- { [ 4-amino-7- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

A suspension of example 125 (88 mg, 194 μmol) in dichloromethane (5ml) was treated with thionyl chloride (29 μ l,389 μmol) and the mixture was stirred at room temperature for 65 min. Another portion of thionyl chloride (29 μ l,389 μmol) was added and stirring was continued for 1.5 h. Excess methanol was then added followed by dropwise addition of a 5.4M solution of sodium methoxide in methanol (84 mg, 1.56 mmol) until pH 8 was reached. After stirring for 3 days, the volatiles were evaporated under reduced pressure and the residue was isolated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous trifluoroacetic acid). The product fractions were evaporated to yield 19mg of the title compound (21% of theory).

Example 127

4- { [ 4-amino-7- (ethoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] methyl } piperazin-2-one

A suspension of example 125 (100 mg, 221 μmol) in dichloromethane (10 ml) was treated with thionyl chloride (161 μ l, 2.21 mmol) and the mixture was stirred at room temperature for 30 min. Ethanol was added and the volatiles were evaporated under reduced pressure. The residue was dissolved in ethanol (10 ml), sodium ethoxide (30 mg, 442 μmol) was added, and the mixture was stirred at room temperature for 1 h. The mixture was then directly separated by preparative RP-HPLC (Reprosil C18, gradient 20-40% acetonitrile/0.2% aqueous trifluoroacetic acid). The product fractions were combined and basified by addition of saturated aqueous sodium bicarbonate. The solution was extracted with ethyl acetate and the organic layer was washed with brine, dried over magnesium sulfate and evaporated to yield 57 mg of the title compound (54% of theory).

B. Evaluation of biological Activity

Abbreviations and acronyms:

ahx 6-aminocaproic acid

ATP adenosine triphosphate

BSA bovine serum albumin

CREB cAMP-responsive element binding proteins

DMSO dimethyl sulfoxide

EDTA ethylene diamine tetraacetic acid

EGTA ethylene glycol-bis (2-aminoethylether) -N,N,N',N'-tetraacetic acid

FBS fetal bovine serum

FGF fibroblast growth factor

FGFR fibroblast growth factor receptor

GFP Green fluorescent protein

GST glutathione S-transferase

HEPES 4- (2-hydroxyethyl) piperazine-1-ethanesulfonic acid

HRTF homogeneous time-resolved fluorescence

MOPS 3-(N-morpholino) propanesulfonic acid

Rapamycin targets in mTOR mammals

PBS phosphate buffered saline

PI3K phosphatidylinositol 3-kinase

RTK receptor tyrosine kinases

SNP single nucleotide polymorphism

TR-FRET time resolved fluorescence resonance energy transfer

VEGF vascular endothelial growth factor

VEGFR vascular endothelial growth factor receptor.

Demonstration of the activity of the compounds of the invention can be accomplished by in vitro, ex vivo and in vivo assays well known in the art. For example, to demonstrate the activity of the compounds of the present invention, the following assays may be used.

B-1.FGFR-1 high ATP kinase assay

FGFR-1 inhibitory activity of the compounds of the invention at high ATP concentrations after pre-incubation with FGFR-1 using a TR-FRET based FGFR-1 high ATP assay quantification, as described in the following paragraphs:

a recombinant tagged FGFR-1 fusion protein [ fusion of glutathione-S-transferase (GST) (N-terminal), His6 tag, thrombin cleavage site, and intracellular portion of amino acids G400 to R800 of human FGFR-1 as GenBank accession No. NM-015850 ] expressed in SF9 insect cells using a baculovirus expression system and purified via glutathione-agarose affinity chromatography was purchased from Proqinase (product No. 0101-0000-1) and used as an enzyme. As substrate for the kinase reaction, the biotinylated peptide biotin-Ahx-AAEEEYFFLFAKKK (C-terminus in amide form) which is commercially available, for example, from Biosyntan (Berlin-Buch, Germany) is used.

Typically, test compounds are tested in duplicate at 11 different concentrations (e.g., 20 μ M, 5.9 μ M, 1.7 μ M, 0.51 μ M, 0.15 μ M, 44 nM, 13 nM, 3.8 nM, 1.1 nM, 0.33 nM, and 0.1nM) in the range of 20 μ M to 0.1nM in the same microtiter plate. Dilution series were prepared as 100-fold concentrated stock solutions in DMSO prior to assay, respectively; the exact concentration may vary depending on the pipette used. For this assay, 50 nl of each stock solution of test compound in DMSO was pipetted into a black, small-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany). A solution of 2 μ l of the above FGFR-1 fusion protein in aqueous assay buffer [ 8mM MAMPS pH 7.0, 10mM magnesium acetate, 1.0 mM dithiothreitol, 0.05% (w/v) Bovine Serum Albumin (BSA),0.07% (v/v) Tween-20, 0.2 mM EDTA ] was added and the mixture was incubated at 22 ℃ for 15min to allow pre-binding of the test compound to the enzyme. Then, the kinase reaction was started by adding 3 μ l of adenosine triphosphate (ATP, 3.3 mM; final concentration in5 μ l assay volume = 2 mM) and a solution of substrate (0.16 μ M; final concentration in5 μ l assay volume = 0.1 μ M) in assay buffer, and the resulting mixture was incubated at 22 ℃ for a reaction time of 15 min. The concentration of FGFR-1 fusion protein was adjusted according to the activity of the enzyme batch and was appropriately selected to have an assay in the linear range (typically in the range of 0.05 μ g/ml concentration). The reaction was stopped by adding 5 μ l of a solution of HTRF detection reagent [25 nM streptavidin-XL 665 (Cis Biointernational) and 1nM PT 66-Eu-chelate, an europium chelate-labeled anti-phosphotyrosine antibody (Perkin-Elmer; can be replaced with PT 66-Tb-cryptate from Cis Biointernational), in aqueous EDTA (50 mM EDTA, 0.1% (w/v) BSA in 50 mM HEPES/NaOH pH 7.5 ].

The resulting mixture was incubated at 22 ℃ for 1 hour to allow the formation of a complex between the phosphorylated biotinylated peptide and the detection reagent. Subsequently, the amount of phosphorylated substrate was evaluated by determining the resonance energy transfer from the Eu-chelate to streptavidin-XL 665. For this purpose, in a TR-FRET reader [ e.g., Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer)]Measured 620 nm and 665 nm fluorescence emission after 350 nm excitation. The emission ratios at 665 nm and 620 nm were used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor = 0% inhibition, all other assay components but no enzyme = 100% inhibition) and IC was calculated by 4-parameter fitting using internal software₅₀The value is obtained.

IC of Compounds of the invention from this assay₅₀Values are listed in table 1A below:

。

synthesis of selected 8-amino-1- (benzothien-2-yl) imidazo [1,5-a ] s, which are considered to be the closest representatives of the prior art (see international patent application WO2007/061737-A2 and the example compounds described therein), following published procedures]Pyrazine derivatives and related compounds, and also tested in FGFR-1 high ATP assays for comparative purposes. To pairIC obtained from these compounds₅₀Values are listed in table 1B below:

。

IC specified in tables 1A and 1B₅₀The values indicate that the compounds of the invention are about 5-fold to one thousand-fold more potent in inhibiting FGFR-1 kinase activity than the selected prior art compounds.

B-2.FGFR-3 kinase assay

FGFR-3 inhibitory activity of the compounds of the invention after pre-incubation with FGFR-3 was quantified using a TR-FRET based FGFR-3 assay as described in the following paragraphs:

recombinant tagged FGFR-3 fusion proteins [ fusion of glutathione-S-transferase (GST) (N-terminus), His6 tag, thrombin cleavage site, and intracellular portion of amino acid R397 of human FGFR-3 as NCBI/Protein accession No. NP _000133.1 to T806 ] expressed in SF9 insect cells using a baculovirus expression system and purified via glutathione-S-transferase affinity chromatography were purchased from Proqinase (product No. 1068-. As substrate for the kinase reaction, the biotinylated peptide biotin-Ahx-AAEEEYFFLFAKKK (C-terminus in amide form) which is commercially available, for example, from Biosyntan (Berlin-Buch, Germany) is used.

Typically, test compounds are tested in duplicate at 11 different concentrations (e.g., 20 μ M, 5.9 μ M, 1.7 μ M, 0.51 μ M, 0.15 μ M, 44 nM, 13 nM, 3.8 nM, 1.1 nM, 0.33 nM, and 0.1nM) in the range of 20 μ M to 0.1nM in the same microtiter plate. Dilution series were prepared as 100-fold concentrated stock solutions in DMSO prior to assay, respectively; the exact concentration may vary depending on the pipette used. For this assay, 50 nl of each stock solution of test compound in DMSO was pipetted into a black, small-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany). A solution of 2 μ l of the above FGFR-3 fusion protein in aqueous assay buffer [ 8mM MAMPS pH 7.0, 10mM magnesium acetate, 1.0 mM dithiothreitol, 0.05% (w/v) Bovine Serum Albumin (BSA),0.07% (v/v) Tween-20, 0.2 mM EDTA ] was added and the mixture was incubated at 22 ℃ for 15min to allow pre-binding of the test compound to the enzyme. Then, the kinase reaction was started by adding 3 μ l of adenosine triphosphate (ATP, 16.7 μ M; final concentration = 10 μ M in5 μ l measurement volume) and a solution of substrate (0.8 μ M; final concentration = 0.5 μ M in5 μ l measurement volume) in the assay buffer, and the resulting mixture was incubated at 22 ℃ for a reaction time of 60 min. The concentration of FGFR-3 fusion protein was adjusted according to the activity of the enzyme batch and was appropriately selected to have an assay in the linear range (typically in the range of 0.03 μ g/ml concentration). The reaction was stopped by adding 5 μ l of a solution of HTRF detection reagent [100 nM streptavidin-XL 665 (Cis Biointernational) and 1nM PT 66-Tb-cryptate, a terbium cryptate-labeled anti-phosphotyrosine antibody (Cis Biointernational; which may be replaced with PT 66-Eu-chelate from Perkin-Elmer), in aqueous EDTA (50 mM EDTA, 0.1% (w/v) BSA in 50 mM HEPES/NaOH pH 7.5 ].

The resulting mixture was incubated at 22 ℃ for 1 hour to allow the formation of a complex between the phosphorylated biotinylated peptide and the detection reagent. Subsequently, the amount of phosphorylated substrate was assessed by determining the resonance energy transfer from the Tb-chelate to streptavidin-XL 665. For this purpose, in a TR-FRET reader [ e.g., Rubystar (BMG Labtechnologies, Offenburg, Germany) or Viewlux (Perkin-Elmer)]Measured 620 nm and 665 nm fluorescence emission after 350 nm excitation. The emission ratios at 665 nm and 620 nm were used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor = 0% inhibition, all other assay components but no enzyme = 100% inhibition) and IC was calculated by 4-parameter fitting using internal software₅₀The value is obtained.

IC of Compounds of the invention from this assay₅₀Values are listed inIn table 2A below:

。

synthesis of selected 8-amino-1- (benzothien-2-yl) imidazo [1,5-a ] s, which are considered to be the closest representatives of the prior art (see international patent application WO2007/061737-A2 and the example compounds described therein), following published procedures]Pyrazine derivatives and related compounds, and also tested in FGFR-3 assays for comparative purposes. IC obtained for these compounds₅₀Values are listed in table 2B below:

。

IC specified in tables 2A and 2B₅₀The values indicate that the compounds of the invention are about three to one thousand times more potent in inhibiting FGFR-3 kinase activity than the selected prior art compounds.

B-3.FGFR-4 high ATP kinase assay

FGFR-4 inhibitory activity of the compounds of the invention at high ATP concentrations following pre-incubation with FGFR-4 using a TR-FRET based FGFR-4 high ATP assay quantification, as described in the following paragraphs:

recombinant tagged FGFR-4 fusion proteins [ fusion of glutathione-S-transferase (GST) (N-terminal), His6 tag, thrombin cleavage site, and intracellular portion of amino acid R391 to T802 of human FGFR-4 as GenBank accession No. NM — 002011 ], expressed in SF9 insect cells using a baculovirus expression system and purified via glutathione-agarose affinity chromatography ] were purchased from Proqinase (product No. 0127-0000-3) and used as enzymes. As substrate for the kinase reaction, the biotinylated peptide biotin-Ahx-AAEEEYFFLFAKKK (C-terminus in amide form) which is commercially available, for example, from Biosyntan (Berlin-Buch, Germany) is used.

Typically, test compounds are tested in duplicate at 11 different concentrations (e.g., 20 μ M, 5.9 μ M, 1.7 μ M, 0.51 μ M, 0.15 μ M, 44 nM, 13 nM, 3.8 nM, 1.1 nM, 0.33 nM, and 0.1nM) in the range of 20 μ M to 0.1nM in the same microtiter plate. Dilution series were prepared as 100-fold concentrated stock solutions in DMSO prior to assay, respectively; the exact concentration may vary depending on the pipette used. For this assay, 50 nl of each stock solution of test compound in DMSO was pipetted into a black, small-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany). A solution of 2 μ l of the above FGFR-4 fusion protein in aqueous assay buffer [ 8mM MAMPS pH 7.0, 10mM magnesium acetate, 1.0 mM dithiothreitol, 0.05% (w/v) Bovine Serum Albumin (BSA),0.07% (v/v) Tween-20, 0.2 mM EDTA ] was added and the mixture was incubated at 22 ℃ for 15min to allow pre-binding of the test compound to the enzyme. Then, the kinase reaction was started by adding 3 μ l of adenosine triphosphate (ATP, 3.3 mM; final concentration in5 μ l assay volume = 2 mM) and a solution of substrate (0.8 μ M; final concentration in5 μ l assay volume = 0.5 μ M) in assay buffer, and the resulting mixture was incubated at 22 ℃ for a reaction time of 60 min. The concentration of FGFR-4 fusion protein was adjusted according to the activity of the enzyme batch and was appropriately selected to have an assay in the linear range (typically in the range of 0.03 μ g/ml concentration). The reaction was stopped by adding 5 μ l of a solution of HTRF detection reagent [100 nM streptavidin-XL 665 (Cis Biointernational) and 1nM PT 66-Tb-cryptate, a terbium cryptate-labeled anti-phosphotyrosine antibody (Cis Biointernational; which may be replaced with PT 66-Eu-chelate from Perkin-Elmer), in aqueous EDTA (50 mM EDTA, 0.1% (w/v) BSA in 50 mM HEPES/NaOH pH 7.5 ].

B-4. For comparison purposesmTOR kinase assay ()

The mTOR inhibitory activity of the compounds of the invention is quantified using a TR-FRET based mTOR assay, as described in the following paragraphs:

recombinant fusion tagged mTOR protein expressed in insect cells and purified by glutathione-agarose affinity chromatography [ glutathione-SFusions of transferase (GST) with amino acids 1360 to 2549 of human mTOR]Purchased from Invitrogen (cat No. 4753) and used as an enzyme. As substrates for the kinase reaction, recombinant fusion proteins of GFP and 4E-BP1 (purchased from Invitrogen, cat. No. PV4759) were used.

Test compounds were dissolved in DMSO to create a 10mM stock solution. These solutions were first diluted 10-fold by 100% DMSO to give 1mM solutions in 100% DMSO, then diluted 100-fold by 50% DMSO to give 10 μ M solutions in 50% DMSO.

For this assay, a 10 μ M solution of test compound in 0.5 μ l 50% DMSO was pipetted into a black small-volume 384-well microtiter plate (Greiner Bio-One, Frickenhausen, Germany). A solution of 2 μ l of the above mTOR fusion protein in aqueous assay buffer [50 mM HEPES/NaOH pH 7.5, 5mM magnesium chloride, 1.0 mM dithiothreitol, 1mM EGTA, 0.01% (v/v) Triton-X100, 0.01% (w/v) Bovine Serum Albumin (BSA) ] was added and the mixture was incubated at 22 ℃ for 15min to allow the test compound to pre-bind to the enzyme. Then, the kinase reaction was started by adding 2.5 μ l of adenosine triphosphate (ATP, 80 μ M; final concentration = 40 μ M in5 μ l measurement volume) and a solution of substrate (0.6 μ M; final concentration = 0.3 μ M in5 μ l measurement volume) in the assay buffer, and the resulting mixture was incubated at 22 ℃ for a reaction time of 60 min. The concentration of mTOR fusion protein was chosen appropriately to have an assay in a linear range (typical final concentration in a 5 μ l assay volume is 1.25 ng/μ l). The reaction was stopped by adding 30 mM EDTA (final concentration = 15 mM in10 μ l assay volume) and 2 nM Tb-chelate labeled anti-4E-BP 1 [ pT46] phosphate specific antibody [ Invitrogen cat # PV4755] (final concentration = 1nM in10 μ l assay volume) in5 μ l FRET buffer.

The resulting mixture was incubated at 22 ℃ for 1 hour to allow for the formation of a complex between the phosphorylated substrate and the Tb-chelate labeled antibody. Subsequently, the amount of phosphorylated substrate was assessed by determining the resonance energy transfer from Tb-chelate to GFP. For this purpose, the fluorescence emissions at 495 nm and 520nm after excitation at 340 nm were measured in an Envision 2104 multiple label reader (Perkin-Elmer). The emission ratios at 520nm and 495 nm were used as a measure of the amount of phosphorylated substrate. Data were normalized (enzyme reaction without inhibitor = 0% inhibition, all other assay components but no enzyme = 100% inhibition) and mean values (if tested repeatedly at a single concentration) or IC were calculated₅₀Values (by 4-parameter fit using internal software).

The average inhibition values at 1 μ M for each compound of the invention are listed in table 3 below:

(no inhibition detected = no inhibition detected at 1 μ M).

The data in table 3 show that the compounds of the invention have only a weak (if any) inhibitory effect on mTOR kinase, which is not believed to contribute to the pharmacological activity observed with these compounds.

B-5.Growth factor-mediated inhibition of cell proliferation

Human Umbilical Vein Endothelial Cells (HUVEC) were obtained from Cellsystems (FC-0003) and were incubated at 37 ℃ and 5% CO₂Next, the cells were grown in Vasculife VEGF complete medium (cells, LL-1020) containing 2% Fetal Bovine Serum (FBS). The cells were used for proliferation assays up to passage 7.

HUVEC cells were harvested using accutase (PAA, L11-007) and seeded at a cell density of 2500 cells/well in 100 μ L Vasculife VEGF complete medium in columns 2-12 in 96 well PLATEs (Falcon MICROTEST tissue culture PLATE 96-well flat bottom, BD 353075, or μ CLEAR-PLATE, black, 96-well, Greiner Bio-One, number 655090), with column 1 left empty as a blank. Cells were incubated at 37 ℃ and 5% CO₂And then incubating for at least 6 h. The cells were then washed once with PBS and starved overnight in vascauline basal medium (cells, LM-0002) containing heparin, ascorbate and L-glutamine (components of the vascauline lifefans kit, cells systems, LL-1020) and 0.2% FBS.

After about 18 h, the starvation medium was discarded and the cells were exposed to 9 consecutive logarithmic or semilogarithmic concentrations of the test compound and 5, 10 or 20 ng/ml of hFGF-2 (recombinant human FGF, basal, R.sub.R.) in the range of 10 pM to 30 μ M in 100 μ l of starvation medium&D Systems, 233-FB) for 72 h. A 10mM stock solution of test compound in DMSO was diluted to 200 x final concentration in DMSO, resulting in a final DMSO concentration of 0.5% in all wells. Controls consisted of cells grown only in starvation medium and consisted of cells withComposition of cells grown in starvation medium containing hFGF-2 in 0.5% DMSO. To determine cell proliferation, 5 μ l of Alamar Blue solution (Biosource, DAL1100) was added to each well (1:20 dilution) and the cells were allowed to stand at 37 ℃ and 5% CO₂After a further 4 h incubation, fluorescence (excitation 535 nm, emission 595 nm) was measured using a Spectrafluor Plus Tecan plate reader (XLUOR 4 version 4.20). In some experiments, an ATP assay kit (BIAFFIN GmbH, LBR-T100) was used according to the manufacturer's instructions. In each experiment, samples were assayed in triplicate and standard deviations were determined. Data were analyzed using GraphPad Prism 5 software and ICs were obtained₅₀The value is obtained. All tested compounds were tested 2-10 times in separate experiments and similar results were obtained.

The data set forth in Table 4 below represent the corresponding average pIC for representative compounds of the invention₅₀Value generated IC₅₀The value:

。

when vascular endothelial growth factor (VEGF-A)₁₆₅Isoforms) used as a mediating growth factor (rather than FGF-2), most compounds of the invention showed about ten to one hundred fold reduced inhibitory activity in this proliferation assay, indicating significant selectivity of these compounds for FGFR compared to VEGFR kinase.

B-6.Human xenograft and syngeneic tumor models

Different tumor models have been performed to analyze the in vivo profile of the compounds of the invention. Human, rat or mouse tumor cells are cultured in vitro and implanted into immunodeficient or immunocompetent mice, or immunodeficient rats. Treatment was initiated after tumor establishment and was treated with the substance via different routes (oral, intravenous, intraperitoneal or subcutaneous)Treating tumor-bearing animals. The substances are tested as monotherapy or in combination therapy with other pharmacological substances. Treating tumor-bearing animal until tumor reaches 120mm²Tumor was measured in two dimensions using calipers, and according to the formula (length × width)²) The tumor volume was calculated 2. Drug efficacy was assessed at the end of the experiment using the T/C ratio [ T = final tumor weight for treatment group; c = final tumor weight of control group]. Statistical significance of efficacy between control and treatment groups was determined using the ANOVA variance test. All animal studies were performed according to the german regulatory guidelines.

Although the present invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and modifications may be devised by those skilled in the art without departing from the true spirit and scope of the present invention. It is intended that the claims be interpreted to include all such embodiments and equivalent variations.

C.Examples relating to pharmaceutical compositions

The pharmaceutical composition of the present invention can be illustrated as follows:

sterile intravenous solution:

sterile, water for injection can be used to prepare a solution of 5mg/mL of the subject compound, and if necessary, the pH adjusted. The solution was diluted with sterile 5% glucose to 1-2 mg/mL for administration and administered as an intravenous infusion over about 60 minutes.

Lyophilized powder for intravenous administration:

can be used (i) 100-1000 mg of the object compound of the invention, (ii) as a lyophilized powderii)32-327 mg/mL sodium citrate and (C)iii)300-3000 mg of dextran 40 was used to prepare the sterile preparation. The formulation is reconstituted with sterile, injectable saline or 5% glucose to a concentration of 10 to 20 mg/mL and further diluted with saline or 5% glucose to 0.2 to 0.4 mg/mL for administration as an intravenous bolus or by intravenous infusion over 15-60 minutes.

Intramuscular suspension:

the following solutions or suspensions can be prepared for intramuscular injection:

50 mg/mL of the water-insoluble compound required in the present invention; 5mg/mL sodium carboxymethylcellulose; 4 mg/ml Tween 80; 9 mg/mL sodium chloride; 9 mg/mL benzyl alcohol.

Hard shell capsule:

a large number of unit capsules were prepared by filling standard two-piece hard gelatin capsules with 100 mg each of the powdered compound required for the present invention, 150 mg of lactose, 50 mg of cellulose and 6mg of magnesium stearate.

Soft capsule preparation:

a mixture of the subject compound in a digestible oil such as soybean oil, cottonseed oil or olive oil is prepared and injected by a positive displacement pump into molten gelatin to form soft gelatin capsules containing 100 mg of the active ingredient. The capsules are washed and dried. The subject compounds of the invention can be dissolved in a mixture of polyethylene glycol, glycerol and sorbitol to prepare a water-miscible drug mixture.

And (3) tablet preparation:

a large number of tablets are prepared by conventional procedures so that the dosage unit is 100 mg of the subject compound, 0.2 mg of colloidal silicon dioxide, 5mg of magnesium stearate, 275 mg of microcrystalline cellulose, 11 mg of starch and 98.8 mg of lactose. Suitable aqueous and non-aqueous coatings may be applied to increase palatability, improve aesthetics and stability, or delay absorption.

Solution or suspension for topical administration to the eye (eye drops):

sterile preparations are prepared with 100 mg of the subject compound in lyophilized powder form reconstituted in 5mL of sterile saline. Benzalkonium chloride, thimerosal, phenylmercuric nitrate, and the like may be used as the preservative in the range of about 0.001% to 1% by weight.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof

Wherein

R¹Is hydrogen, chlorine, methyl or methoxy,

provided that R is¹And R²At least one of which is not hydrogen,

G¹represents chlorine, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkoxycarbonyl, oxazolyl or a radical-CH₂-OR³or-CH₂-NR⁴R⁵，

Wherein

R³Is hydrogen, (C)₁-C₄) Alkyl, unsubstituted (C)₃-C₆) -a cycloalkyl group or an unsubstituted phenyl group,

(i) said (C)₁-C₄) Alkyl optionally substituted by hydroxy, (C)₁-C₄) -alkoxy, hydroxycarbonyl, (C)₁-C₄) Alkoxycarbonyl, amino, aminocarbonyl, (C)₃-C₆) -cycloalkyl or up to three fluorine atoms,

R⁴is hydrogen or (C)₁-C₄) -an alkyl group,

R⁵is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkylcarbonyl, unsubstituted (C)₃-C₆) -cycloalkyl or oxopyrrolidinyl, wherein

(i) Said (C)₁-C₄) Alkyl optionally substituted by hydroxy, hydroxycarbonyl, or (C)₃-C₆) -a cycloalkyl group substitution,

or

R⁴And R⁵Are linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 7-membered heterocycloalkyl ring which may contain a ring selected from N (R)⁷) And a second ring heteroatom of O, and may be independently selected from (C) on a ring carbon atom₁-C₄) -one or two substituents of alkyl, oxo, hydroxy, amino and aminocarbonyl, and wherein

R⁷Is a hydrogen atom, and is,

and is

G²Represents chlorine, cyano, (C)₁-C₄) -alkyl or a radical-CR^8AR^8B-OH、-CH₂-NR⁹R¹⁰、-C(＝O)-NR¹¹R¹²or-CH₂-OR¹⁵Wherein

R^sAAnd R^8BIndependently selected from hydrogen, (C)₁-C₄) -alkyl, cyclopropyl and cyclobutyl,

R⁹is hydrogen or (C)₁-C₄) -an alkyl group,

R¹⁰is hydrogen, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkylcarbonyl, unsubstituted (C)₃-C₆) -cycloalkyl or oxopyrrolidinyl, wherein

(i) Said (C)₁-C₄) -alkyl is optionally substituted by aminocarbonyl,

R¹¹is hydrogen or (C)₁-C₄) -an alkyl group,

R¹²is hydrogen, (C)₁-C₄) Alkyl, unsubstituted (C)₃-C₆) -cycloalkyl or oxopyrrolidinyl, wherein

or

R⁹And R¹⁰Or R is¹¹And R¹²Are each linked and together with the nitrogen atom to which they are attached form a monocyclic saturated 4-to 7-membered heterocycloalkyl ring which may contain a ring selected from N (R)¹³) O, S and S (O)₂And may be independently selected from fluorine, (C) on a ring carbon atom₁-C₄) -alkyl, oxo, hydroxy, amino and aminocarbonyl and wherein

R¹³Is hydrogen, (C)₃-C₆) -cycloalkyl, formyl or (C)₁-C₄) -an alkyl-carbonyl group,

and is

R¹⁵Is (C)₁-C₄) -an alkyl group,

with the proviso that when G²When it is chloro or cyano, G¹Is not chlorine.

2. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein

R¹Is a chlorine, a methyl or a methoxy group,

G¹represents chlorine, (C)₁-C₄) Alkyl radicals, (C)₁-C₄) -alkoxycarbonyl or oxazolyl, or represents the group-CH₂-OR³or-CH₂-NR⁴R⁵Wherein

R³Is hydrogen, (C)₁-C₄) Alkyl or unsubstituted (C)₃-C₆) -a cycloalkyl group,

R⁴is hydrogen or (C)₁-C₄) -an alkyl group,

or

R⁷Is a hydrogen atom, and is,

and is

R^8AAnd R^8BIndependently selected from hydrogen, (C)₁-C₄) -an alkyl group and a cyclopropyl group,

R⁹is hydrogen or a methyl group, or a mixture thereof,

(i) Said (C)₁-C₄) -alkyl optionally substituted by aminocarbonyl

R¹¹Is hydrogen or a methyl group, or a mixture thereof,

(i) Said (C)₁-C₄) -alkyl optionally substituted by hydroxy

Or

R¹³Is hydrogen, cyclopropyl, cyclobutyl, formyl or (C)₁-C₄) -an alkyl-carbonyl group,

and is

R¹⁵Is a methyl group or an ethyl group,

with the proviso that when G²When it is chloro or cyano, G¹Is not chlorine.

3. A compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 1 or 2, wherein

R¹Is a methyl group, and the compound is,

R²is a methoxy group, and the compound is a methoxy group,

R³Is hydrogen, (C)₁-C₄) -an alkyl, cyclopropyl or cyclobutyl group,

R⁴is hydrogen, a methyl group or an ethyl group,

or

and is

G²Represents methyl or a radical-CR^8AR^8B-OH、-CH₂-NR⁹R¹⁰or-C (═ O) -NR¹¹R¹²Wherein

R^8AAnd R^8BIndependently of one another is hydrogen or a methyl group,

R⁹is a hydrogen atom, and is,

R¹¹is hydrogen or a methyl group, or a mixture thereof,

R¹²is hydrogen, (C)₁-C₄) -an alkyl group,Cyclopropyl, cyclobutyl or 2-oxopyrrolidin-3-yl,

or

R¹³Is hydrogen, formyl or acetyl.

4. A compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 1 or 2, wherein

R¹Is a methyl group, and the compound is,

R²is a methoxy group, and the compound is a methoxy group,

G¹represents the group-CH₂-OR³Wherein

and is

G²Represents the group-CH₂-NR⁹R¹⁰or-C (═ O) -NR¹¹R¹²Wherein

R⁹Is a hydrogen atom, and is,

R¹⁰is a 2-oxopyrrolidin-3-yl group,

or

R¹¹is a hydrogen atom, and is,

R¹²is a 2-oxopyrrolidin-3-yl group,

or

R¹¹And R¹²Are connected and connected to themThe attached nitrogen atoms together form a 3-hydroxyazetidin-1-yl, 4-hydroxypiperidin-1-yl or 3-oxopiperazin-1-yl ring.

5. A compound according to claim 1 or 2 which is 4- { [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

Or a pharmaceutically acceptable salt thereof.

6. A compound according to claim 1 or 2 which is 4- { [ 4-amino-6- (methoxymethyl) -5- (7-methoxy-5-methyl-1-benzothien-2-yl) pyrrolo [2,1-f ] [1,2,4] triazin-7-yl ] methyl } piperazin-2-one

7. Process for the preparation of compounds of formula (I) according to any one of claims 1 to 6, characterized in that

[A] 6-substituted 4-aminopyrrolo [2,1-f ] [1,2,4] triazines of formula (II)

Wherein R is³Having the meaning as claimed in any of claims 1 to 4,

Wherein R is⁹And R¹⁰Having the meaning as claimed in any of claims 1 to 4,

to give a compound of the formula (IV)

Wherein R is³、R⁹And R¹⁰Having the meaning as claimed in any of claims 1 to 4,

then brominated to the compound of formula (V)

Wherein R is¹And R²Having the meaning as claimed in any of claims 1 to 4,

and is

R¹⁴Represents hydrogen or (C)₁-C₄) -alkyl, or two R¹⁴The residues are linked together to form- (CH)₂)₂-、-C(CH₃)₂-C(CH₃)₂-、-(CH₂)₃-、-CH₂-C(CH₃)₂-CH₂-or-C (═ O) -CH₂-N(CH₃)-CH₂-a bridge of-C (═ O) -,

to obtain the target compound of formula (I-A)

Wherein R is¹、R²、R³、R⁹And R¹⁰Having the meaning as claimed in any of claims 1 to 4,

or

[B] 6-substituted 4-aminopyrrolo [2,1-f ] [1,2,4] triazines of formula (II)

Wherein R is³Having the meaning as claimed in any of claims 1 to 4,

first formylated with N, N-dimethylformamide in the presence of phosphorus oxychloride to give the aldehyde of the formula (VII)

Wherein R is³Having the meaning as claimed in any of claims 1 to 4,

then brominated to the compound of formula (VIII)

Wherein R is³Having the meaning as claimed in any of claims 1 to 4,

Wherein R is¹、R²And R¹⁴Has the meaning of the above-mentioned formula,

to give a compound of formula (IX)

Wherein R is¹、R²And R3 has the meaning as claimed in any of claims 1 to 4,

then put it in

Wherein R is⁹And R¹⁰Having the meaning as claimed in any of claims 1 to 4,

to obtain the target compound of formula (I-A)

Wherein R is¹、R²、R³、R⁹And R¹⁰Has the meaning as defined in any one of claims 1 to 4, or

[ B-2] Oxidation to carboxylic acids of formula (X)

Wherein R is¹、R²And R³Having the meaning as claimed in any of claims 1 to 4,

Wherein R is¹¹And R¹²Having the meaning as claimed in any of claims 1 to 4,

to obtain the target compound of formula (I-B)

Wherein R is¹、R²、R³、R¹¹And R¹²Having the structure of any one of claims 1 to 4Has the meaning of, or

Wherein R is¹、R²And R¹⁴Has the meaning of the above-mentioned formula,

to give a compound of the formula (XIII)

Wherein R is¹And R²Having the meaning as claimed in any of claims 1 to 4,

Wherein R is⁹And R¹⁰Having the meaning as claimed in any of claims 1 to 4,

to give a compound of the formula (I-C)

Wherein R is¹、R²、R⁹And R¹⁰Having the meaning as claimed in any of claims 1 to 4, which is subsequently reacted

[ C-1] Oxidation to aldehydes of formula (XIV)

Wherein R is¹、R²、R⁹And R¹⁰Has the meaning of any one of claims 1 to 4 and is treated with an amine of the formula (XV) in the presence of an acid and a reducing agent

Wherein R is⁴And R⁵Having the meaning as claimed in any of claims 1 to 4,

to obtain the target compound of formula (I-D)

Wherein R is¹、R²、R⁴、R⁵、R⁹And R¹⁰Has the meaning as defined in any one of claims 1 to 4, or

[ C-2] to the corresponding 6- (halomethyl) derivative of formula (XVI)

Wherein R is¹、R²、R⁹And R¹⁰Having the meaning as claimed in any of claims 1 to 4,

and is

X is chlorine, bromine or iodine,

and treatment with an alcohol of formula (XVII) in the presence of a base

R^3A-OH (XVII)，

Wherein R is^3AHaving the R of any one of claims 1-4³The meaning of (a) is, in addition to hydrogen,

to obtain the target compound of formula (I-E)

the compounds of the formula (I) thus obtained are then optionally, if appropriate, separated into their respective enantiomers and/or diastereomers.

8. Use of a compound according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment and/or prophylaxis of a proliferative disease.

9. Use of a compound according to any one of claims 1 to 6 in the manufacture of a medicament for the treatment and/or prevention of a fibroblast growth factor receptor mediated disease.

10. Use of a compound according to any one of claims 1 to 6 for the preparation of a pharmaceutical composition for the treatment and/or prophylaxis of tumor diseases.

11. The use of claim 10, wherein the neoplastic disease is cancer.

12. A pharmaceutical composition comprising a compound of any one of claims 1-6 and one or more pharmaceutically acceptable excipients.

13. The pharmaceutical composition of claim 12, further comprising one or more additional therapeutic agents.

14. Use of a pharmaceutical composition according to claim 12 or 13 for the preparation of a medicament for the treatment and/or prevention of a tumor disease.

15. The use of claim 14, wherein the neoplastic disease is cancer.