JP2009504692A - Novel 4-amino-thieno [3,2-c] pyridine-7-carboxylic acid amide - Google Patents

Abstract

The present invention relates to novel 4-amino-thieno [3,2-c] pyridine-7-carboxylic acid amides of formula (I) and pharmaceutically acceptable salts and esters thereof. The compounds of formula (I) are selective inhibitors of KDR kinase and / or FGFR kinase. These compounds and their pharmaceutically acceptable salts are antiproliferative agents useful for the treatment or control of solid tumors, particularly solid tumors of the breast, colon, lung, and prostate. Also disclosed are pharmaceutical compositions or medicaments containing these compounds, methods for producing these compounds, and methods for treating cancer using these compounds.

Description

  The present invention relates to 4-amino-thieno [3,2-c] pyridine-7-carboxylic acid amides and pharmaceutically acceptable salts and esters thereof. These compounds inhibit KDR (kinase insertion domain containing receptor) kinase and / or FGFR (fibroblast growth factor receptor) kinase. These compounds and their pharmaceutically acceptable salts and esters have antiproliferative activity and are useful in the treatment or control of cancer, specifically solid tumors. In addition, these compounds have an advantageous bioavailability profile. The invention also relates to pharmaceutical compositions containing such compounds, and also to methods for treating or controlling cancer, most preferably methods for treating or controlling breast cancer, lung cancer, colon cancer, or prostate cancer.

  Protein kinases are a group of proteins (enzymes) that regulate various cellular functions. This is accomplished by causing structural changes in the substrate protein as a result of phosphorylation of specific amino acids on the protein substrate. This structural change modulates the activity of the substrate or the ability of the substrate to interact with other binding partners. The enzymatic activity of a protein kinase refers to the rate at which the kinase adds a phosphate group to a substrate. The measurement can be performed, for example, by detecting the amount of substrate and converting it to a product that is a function of time. Substrate phosphorylation occurs at the active site of protein kinases.

  Tyrosine kinases are a subset of protein kinases that catalyze the transfer of the terminal phosphate group of adenosine triphosphate (ATP) to a tyrosine residue on a protein substrate. These kinases play an important role in the propagation of growth factor signaling leading to cell proliferation, differentiation and migration.

  For example, basic fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) have been recognized as important mediators of tumor-promoting angiogenesis. VEGF activates endothelial cells by signaling through two high affinity receptors, one of which is the kinase insert domain containing receptor (KDR). See Hennequin L. F. et al., J. Med. Chem. 2002, 45 (6), pp1300. FGF activates endothelial cells by signaling through the FGF receptor (FGFR). Solid tumor growth depends on the formation of new blood vessels (angiogenesis). Thus, inhibitors of the receptors FGFR and / or KDR that interfere with growth signaling and thus slow or prevent angiogenesis are useful agents in the prevention and treatment of solid tumors. See Klohs W. E. et al., Current Opinion in Biotechnology 1999, 10, p.544.

  In order to treat one or more solid tumors, there is a need for easily synthesized small molecule compounds that effectively inhibit the catalytic activity of protein kinases (particularly FGFR and KDR kinases). Particularly desirable is the provision of small molecule inhibitors that are selective for FGFR and / or KDR. This is because inhibition of multiple targets can cause potential incidental toxicity and other undesirable complications. Such small molecule inhibitors also preferably have an advantageous bioavailability profile.

  Accordingly, an object of the present invention is to provide such compounds and pharmaceutical compositions containing these compounds.

の化合物、又は医薬的に許容可能なその塩又はエステルに関する。ここで、Ｒ¹及びＲ²は、後に定義する通りである。 According to one embodiment, the present invention provides a novel 4-amino-thieno [3,2-c] pyridine-7-carboxylic acid amide capable of selectively inhibiting the activity of KDR and / or FGFR. About. These compounds are useful for the treatment or control of cancer, specifically the treatment or control of solid tumors. In particular, the present invention provides a formula

Or a pharmaceutically acceptable salt or ester thereof. Here, R ¹ and R ² are as defined later.

  The present invention also includes a pharmaceutical composition comprising a therapeutically effective amount of one or more compounds of Formula I, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier or excipient. About.

  The present invention further provides a method of treating or controlling solid tumors, in particular breast, by administering a therapeutically effective amount of a compound of formula I and / or a pharmaceutically acceptable salt thereof to a human patient in need of such treatment. It relates to a method for the treatment or control of tumors, lung tumors, colon tumors or prostate tumors, in particular breast tumors or colon tumors.

  The present invention further relates to novel intermediate compounds useful for the preparation of compounds of formula I.

  As used herein, the following terms shall have the following definitions.

“Alkyl” means a straight or branched saturated aliphatic hydrocarbon having 1 to 10, preferably 1 to 6, more preferably 1 to 4 carbon atoms. An alkyl group having 1 to 6 carbon atoms is also referred to herein as “lower alkyl”. Typical lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, t-butyl, 2-butyl, pentyl, and hexyl. As used herein, for example, notation C _{1 ~ 4} alkyl refers to alkyl having 1 to 4 carbon atoms.

  “Aryl” means an aromatic carbocyclic group, for example a 6-10 membered aromatic or partially aromatic ring system. Preferred aryl groups include, but are not limited to, phenyl, naphthyl, tolyl, and xylyl.

  “Cycloalkyl” means a non-aromatic moiety of 3 to 8 carbon atoms or a fully saturated cycloaliphatic hydrocarbon group. Examples of cycloalkyl groups include cyclopropyl, cyclopentyl, and cyclohexyl.

  “Effective amount” or “therapeutically effective amount” means an amount of at least one compound of Formula I, or a pharmaceutically acceptable salt thereof, that significantly inhibits tumor growth.

  “Halogen” means fluorine, chlorine, bromine, or iodine, preferably bromine, chlorine, or fluorine.

“Hetero atom” means an atom selected from N, O and S. Of these, N is preferable. When the heteroatom is N, it may be present as -NH- or -N-lower alkyl-. If the heteroatom is S, S, may be present SO, or as SO _2.

  “Heteroaryl” means an aromatic heterocyclic ring system containing up to two rings. Preferred heteroaryl groups include, but are not limited to, thienyl, furyl, indolyl, pyrrolyl, pyridinyl, pyrazinyl, oxazolyl, thiaxolyl, quinolinyl, pyrimidinyl, imidazolyl, and tetrazolyl.

  "Heterocycle" or "heterocyclyl" is a 3-10 membered saturated or partially unsaturated non-aromatic monovalent having 1-3 heteroatoms selected from nitrogen, oxygen, or sulfur, or combinations thereof Means a cyclic group. Examples of preferred heterocycles include piperidine, piperazine, pyrrolidine, and morpholine.

“IC ₅₀ ” means the concentration of a specific compound according to the present invention that is necessary to inhibit 50% of the specific activity measured. IC ₅₀ measurements can be made, inter alia, as described in Example 26 below.

A “pharmaceutically acceptable ester” is a compound of formula I having a carboxyl group, esterified by conventional methods, wherein the ester retains the biological effectiveness and properties of the compound of formula I. , A compound that is cleaved in vivo to yield the corresponding active carboxylic acid. Examples of ester groups that are cleaved in vivo (hydrolysis in this case) to give the corresponding carboxylic acid (R ⁴⁰ C (═O) OH) include lower alkyl esters (optionally substituted with NR ⁴¹ R ⁴² ) Here, R ⁴¹ and R ⁴² are lower alkyl, or NR ⁴¹ R ⁴² is united to form a monocyclic aliphatic heterocyclic ring such as pyrrolidine, piperidine, morpholine, N-methylpiperazine, etc.) ; acyloxyalkyl esters of the formula ^{R 40 C (= O) OCHR} 43 OC (= O) R 44 ( wherein, R ⁴³ is hydrogen or methyl, R ⁴⁴ is lower alkyl or cycloalkyl); wherein R ⁴⁰ Carbonate ester of C (= O) OCHR ⁴³ OC (= O) R ⁴⁵ where R ⁴³ is hydrogen or methyl and R ⁴⁵ is lower alkyl or cycloalkyl; formula R ⁴⁰ C (= O) OC ₂ C (= O) aminocarbonyl methyl ester of NR ⁴¹ R ⁴² (wherein, either R ⁴¹ and R ⁴² is hydrogen or lower alkyl, or NR ⁴¹ R ⁴² together form a monocyclic aliphatic heterocycle A ring such as pyrrolidine, piperidine, morpholine, N-methylpiperazine, etc.).

Examples of lower alkyl esters include methyl, ethyl, and n-propyl esters. Examples of the lower alkyl ester substituted with NR ⁴¹ R ⁴² include diethylaminoethyl, 2- (4-morpholinyl) ethyl, 2- (4-methylpiperazin-1-yl) ethyl ester, and the like. Examples of the acyloxyalkyl ester include pivaloxymethyl, 1-acetoxyethyl, and acetoxymethyl ester. Examples of the carbonic acid ester include 1- (ethoxycarbonyloxy) ethyl and 1- (cyclohexyloxycarbonyloxy) ethyl ester. Examples of aminocarbonylmethyl ester include N, N-dimethylcarbamoylmethyl and carbamoylmethyl ester.

  Examples of esters for the delivery of pharmaceutical compounds and further information regarding their use are available from Design of Prodrugs, Bundgaard Hed. (Elsevier, 1985). H. Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems (6th Ed. 1995), pp. 108-109; Krogsgaard-Larsen et al., Textbook of Drug Design and Development (2nd Ed. 1996), pp. See also .152-191.

  “Pharmaceutically acceptable salts” are conventional acid addition or base addition salts that retain the biological effectiveness and properties of the compounds of Formula I and that are suitable non-toxic organic or inorganic acids. Or the thing formed from an organic or inorganic base is meant. Examples of acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid, and p-toluenesulfonic acid, salicylic acid, And salts derived from organic acids such as methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, and fumaric acid. Examples of base addition salts include ammonium hydroxide, potassium hydroxide, sodium hydroxide, and quaternary ammonium hydroxide (eg, tetramethylammonium hydroxide). Chemical modification of pharmaceutical compounds (ie drugs) to salts is a technique well known to medicinal chemists in improving the physical and chemical stability, hygroscopicity, flowability, and solubility of compounds. . See, for example, H. Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems (6th Ed. 1995), pp. 196 and 1456-1457.

  “Pharmaceutically acceptable” such as a pharmaceutically acceptable carrier, excipient, etc. is pharmacologically acceptable and substantially non-toxic to the patient to whom the particular compound is administered. Means that.

  “Substituted” such as substituted alkyl means that substitution may occur at one or more positions, and unless otherwise noted, the substituents at each substitution site are independently designated. It means to be selected from the selected options.

の化合物
（式中、
Ｒ¹は、低級アルキル、及び
ＯＲ³、ＮＲ³Ｒ⁴、Ｓ（Ｏ）_nＲ³、シクロアルキル、置換型シクロアルキル、複素環、置換型複素環、ヘテロアリール、又は置換型ヘテロアリールで置換された低級アルキル
から選択され；
Ｒ²は、
Ｈ、
低級アルキル、及び
ＯＲ⁵、ＯＣ（Ｏ）Ｒ⁵、ＮＲ⁵Ｒ⁶、Ｓ（Ｏ）_nＲ⁵、アリール、置換型アリール、シクロアルキル、置換型シクロアルキル、複素環、置換型複素環、ヘテロアリール、又は置換型ヘテロアリールで置換された低級アルキル
から選択され；
Ｒ³及びＲ⁴は独立して、
Ｈ、
低級アルキル、
アリール、複素環又は置換型複素環に縮合したアリール、置換型アリール、ヘテロアリール、置換型ヘテロアリール、シクロアルキル、置換型シクロアルキル、複素環、又は置換型複素環で置換された低級アルキル、
アリール、
複素環又は置換型複素環に縮合したアリール、
置換型アリール、
ヘテロアリール、
複素環又は置換型複素環に縮合したヘテロアリール、
置換型ヘテロアリール、
複素環、
アリールに縮合した複素環、
シクロアルキル、及び置換型シクロアルキル
から選択され、
或いは、基ＮＲ³Ｒ⁴は独立して、総原子数３〜７の環を形成していてもよく、前記環原子は、Ｒ³及びＲ⁴が結合する窒素に加えて、炭素環原子を含んでなり、前記炭素環原子は任意に、１又は２以上の更なるヘテロ原子によって置換されていてもよく、前記環原子は任意に、１又は２以上の低級アルキル、＝Ｏ、ＯＲ⁷、ＣＯＲ⁷、ＣＯ₂Ｒ⁷、ＣＯＮＲ⁷Ｒ⁸、ＳＯ_nＲ⁷、及びＳＯ₂ＮＲ⁷Ｒ⁸からなる基によって置換されていてもよく；
Ｒ⁵及びＲ⁶は独立して、
Ｈ、
低級アルキル、及び
ＯＲ⁷、ＮＲ⁷Ｒ⁸、アリール、置換型アリール、ヘテロアリール、置換型ヘテロアリール、複素環、シクロアルキル、置換型シクロアルキルで置換された低級アルキル
から選択され、
或いは、基ＮＲ⁵Ｒ⁶は独立して、総原子数３〜７の環を形成していてもよく、前記環原子は、Ｒ⁵及びＲ⁶が結合する窒素に加えて、炭素環原子を含んでなり、前記炭素環原子は任意に、１又は２以上の更なるヘテロ原子によって置換されていてもよく、前記環原子は任意に、１又は２以上の低級アルキル、＝Ｏ、ＯＲ⁷、ＮＲ⁷Ｒ⁸、ＣＯＲ⁷、ＣＯ₂Ｒ⁷、ＣＯＮＲ⁷Ｒ⁸、ＳＯ_nＲ⁷、及びＳＯ₂ＮＲ⁷Ｒ⁸からなる基によって置換されていてもよく；
Ｒ⁷及びＲ⁸は独立して、
Ｈ、及び
低級アルキル
から選択され、或いは、
該基ＮＲ⁷Ｒ⁸は独立して、総原子数３〜７の環を形成していてもよく、前記環原子は、Ｒ⁷及びＲ⁸が結合する窒素に加えて、炭素環原子を含んでなり、前記炭素環原子は任意に、１又は２以上の更なるヘテロ原子によって置換されていてもよく、前記環原子は任意に、１又は２以上の低級アルキル、＝Ｏ、又はＯＲ⁹からなる基によって置換されていてもよく；
Ｒ⁹はＨ、又は低級アルキルであり；
ｎは、０、１、又は２であり；
ここで、
置換型アリール及び置換型ヘテロアリールは、低級アルキル、ＯＲ⁷、ＮＲ⁷Ｒ⁸、ＣＯＲ⁷、ＣＯ₂Ｒ⁷、ＣＯＮＲ⁷Ｒ⁸、ＳＯ₂ＮＲ⁷Ｒ⁸、ＳＯ_nＲ⁷、ＣＮ、ＮＯ₂、及びハロゲンから独立して選択された１又は２以上の基で置換されたアリール及びヘテロアリールであり；
置換型シクロアルキル及び置換型複素環は、低級アルキル、＝Ｏ、ＯＲ⁷、ＮＲ⁷Ｒ⁸、ＣＯＲ⁷、ＣＯ₂Ｒ⁷、ＣＯＮＲ⁷Ｒ⁸、ＳＯ₂ＮＲ⁷Ｒ⁸、ＳＯ_nＲ⁷、及びＣＮから独立して選択された１又は２以上の基で置換されたシクロアルキル及び複素環である）、又は
医薬的に許容可能なその塩又はエステルに関する。 According to one embodiment, the present invention provides a formula

A compound of the formula
R ¹ is substituted with lower alkyl and OR ³ , NR ³ R ⁴ , S (O) _n R ³ , cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, heteroaryl, or substituted heteroaryl Selected from lower alkyl
R ² is
H,
Lower alkyl, and ^{OR 5, OC (O) R} 5, NR 5 R 6, S (O) n R 5, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, Selected from aryl, or lower alkyl substituted with substituted heteroaryl;
R ³ and R ⁴ are independently
H,
Lower alkyl,
Aryl, aryl fused to a heterocycle or substituted heterocycle, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, or lower alkyl substituted with a substituted heterocycle,
Aryl,
Aryl fused to a heterocycle or substituted heterocycle,
Substituted aryl,
Heteroaryl,
Heteroaryl fused to a heterocycle or substituted heterocycle,
Substituted heteroaryl,
Heterocycle,
A heterocycle fused to an aryl,
Selected from cycloalkyl, and substituted cycloalkyl;
Alternatively, the group NR ³ R ⁴ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ³ and R ⁴ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally having one or more lower alkyl, ═O, OR ⁷ , Optionally substituted by a group consisting of COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ ;
R ⁵ and R ⁶ are independently
H,
Selected from lower alkyl and lower alkyl substituted with OR ⁷ , NR ⁷ R ⁸ , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl,
Alternatively, the group NR ⁵ R ⁶ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ⁵ and R ⁶ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally having one or more lower alkyl, ═O, OR ⁷ , Optionally substituted by a group consisting of NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ ;
R ⁷ and R ⁸ are independently
Selected from H and lower alkyl, or
The group NR ⁷ R ⁸ may independently form a ring having 3 to 7 total atoms, and the ring atom contains a carbon ring atom in addition to the nitrogen to which R ⁷ and R ⁸ are bonded. Wherein the carbon ring atom is optionally substituted by one or more additional heteroatoms, and the ring atom is optionally from one or more lower alkyl, ═O, or OR ^9. Optionally substituted by the group
R ⁹ is H or lower alkyl;
n is 0, 1, or 2;
here,
Substituted aryl and substituted heteroaryl, lower ^{alkyl, OR 7, NR 7 R 8} , COR 7, CO 2 R 7, CONR 7 R 8, SO 2 NR 7 R 8, SO n R 7, CN, NO 2 And aryl and heteroaryl substituted with one or more groups independently selected from halogen;
Substituted cycloalkyl and substituted heterocycle are lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO ₂ NR ⁷ R ⁸ , SO _n R ⁷ , And cycloalkyl and heterocycle substituted with one or more groups independently selected from CN), or a pharmaceutically acceptable salt or ester thereof.

  The compounds disclosed herein and the compounds encompassed by Formula I above may exhibit tautomerism or structural isomerism. The present invention includes any tautomeric or structural isomers of these compounds, or mixtures of such forms (eg, racemic mixtures), and any one of the tautomers shown in Formula I above. It is not limited to mutants or structural isomers.

As will be apparent to those skilled in the art, the above NR ³ R ⁴ , NR ⁵ R ⁶ , and NR ⁷ R ⁸ may have one or more ring heteroatoms in addition to the above N. . The total number of additional ring heteroatoms, excluding N described above, depends on the type of ring system involved, but it is preferred that the additional ring heteroatoms do not exceed 1 or 2.

According to one embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with OR ³ . Preferred R ³ groups include aryl, halogen-substituted aryl, and aryl fused to a heterocycle. Preferred halogen groups include Br, Cl, and F.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with OR ³ . Preferred R ³ groups include heteroaryl and heteroaryl substituted with OR ⁷ .

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with NR ³ R ⁴ . The group NR ³ R ⁴ preferably forms a ring with a total of 3 to 7 ring atoms including carbon ring atoms in addition to the nitrogen to which R ³ and R ⁴ are bonded, where the carbon ring atom is optional Optionally substituted by one or more further heteroatoms, wherein said ring atom is optionally one or more lower alkyl, ═O, OR ⁷ , COR ⁷ , CO ₂ R ⁷ , CONR ^7. It may be substituted by a group consisting of R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ , preferably OR ⁷ . Among them, the ring atom is preferably unsubstituted or substituted with lower alkyl and ═O.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with S (O) _n R ³ . Here, R ³ is lower alkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with substituted cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with a heterocycle.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with a substituted heterocycle.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ¹ is lower alkyl substituted with substituted heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ¹ is lower alkyl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with OR ⁵ . Where R ⁵ is lower alkyl substituted with NR ⁷ R ⁸ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with NR ⁵ R ⁶ . Preferably, the group NR ⁵ R ⁶ forms a ring with a total number of 3-7 atoms, said ring atom comprising a carbon ring atom in addition to the nitrogen to which R ⁵ and R ⁶ are attached, said carbon Ring atoms may optionally be substituted by one or more further heteroatoms, said ring atoms optionally having one or more lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , Substituted by a group consisting of COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ . Among these, the ring atom is preferably unsubstituted or substituted by lower alkyl, ═O, and OR ⁷ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ² is lower alkyl substituted with one or more OH groups or one NR ⁵ R ⁶ group.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with OR ⁵ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with OC (O) R ⁵ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with S (O) _n R ⁵ . Here, R ⁵ is lower alkyl, and n is 1 or 2.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with aryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with substituted aryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ² is lower alkyl substituted with substituted cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with a heterocycle.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ² is lower alkyl substituted with a substituted heterocycle.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is lower alkyl substituted with substituted heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ² is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ³ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ³ is lower alkyl.

According to another embodiment, the present invention provides an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, wherein R ³ is fused to an aryl, heterocycle or substituted heterocycle, It relates to compounds of the formula I which are heterocycles or lower alkyl substituted by substituted heterocycles.

According to another embodiment, the present invention relates to a compound of formula I wherein R ³ is aryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is aryl fused to a heterocycle or substituted heterocycle.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is substituted aryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is heteroaryl fused to a heterocycle or substituted heterocycle.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is substituted heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ³ is a heterocycle.

According to another embodiment, the present invention relates to a compound of formula I wherein R ³ is a heterocycle fused to aryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is substituted cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ³ is lower alkyl, heterocycle fused to aryl, aryl, substituted aryl, or aryl fused to heterocycle.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁴ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁴ is lower alkyl.

According to another embodiment, the invention provides an aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, wherein R ⁴ is fused to an aryl, heterocycle or substituted heterocycle, It relates to compounds of the formula I which are heterocycles or lower alkyl substituted by substituted heterocycles.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁴ is aryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ⁴ is aryl fused to a heterocycle or substituted heterocycle.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁴ is substituted aryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ⁴ is heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁴ is heteroaryl fused to a heterocycle or substituted heterocycle.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ⁴ is substituted heteroaryl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁴ is a heterocycle.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ⁴ is a heterocycle fused to aryl.

According to another embodiment, the present invention relates to a compound of formula I, wherein R ⁴ is cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁴ is substituted cycloalkyl.

According to another embodiment, the present invention relates to a compound of formula I, wherein the group NR ³ R ⁴ forms a ring with a total number of atoms of 3-7. Wherein the ring atom comprises a carbon ring atom in addition to the nitrogen to which R ³ and R ⁴ are attached, the carbon ring atom optionally substituted by one or more further heteroatoms. The ring atom may optionally be one or more lower alkyl, ═O, OR ⁷ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ^8. May be substituted by a group consisting of Most preferably, the ring atom is unsubstituted or substituted with lower alkyl, ═O, and OR ⁷ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁵ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁵ is lower alkyl.

According to another embodiment, the present invention provides a lower R ⁵ substituted with NR ⁷ R ⁸ , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl. Relates to compounds of formula I which are alkyl. Most preferably R ⁵ is lower alkyl substituted with NR ⁷ R ⁸ .

According to another embodiment, the present invention relates to a compound of formula I, wherein R ⁵ is lower alkyl substituted by one or more OR ⁷ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁶ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁶ is lower alkyl.

According to another embodiment, the present invention relates to a lower moiety wherein R ⁶ is substituted with NR ⁷ R ⁸ , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, cycloalkyl, substituted cycloalkyl. Relates to compounds of formula I which are alkyl. Among them, R ⁶ is most preferably lower alkyl substituted with NR ⁷ R ⁸ .

According to another embodiment, the present invention relates to a compound of formula I, wherein the group NR ⁵ R ⁶ forms a ring with a total number of atoms of 3-7. Wherein the ring atom comprises a carbon ring atom in addition to the nitrogen to which R ⁵ and R ⁶ are attached, the carbon ring atom optionally substituted by one or more additional heteroatoms. The ring atom may optionally be one or more lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO _{2 It} may be substituted by a group consisting of NR ⁷ R ⁸ . Most preferably, the ring atom is unsubstituted or substituted with lower alkyl, ═O, and OR ⁷ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁷ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁷ is lower alkyl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁸ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁸ is lower alkyl.

According to another embodiment, the present invention relates to a compound of formula I wherein the group NR ⁷ R ⁸ forms a ring with a total number of 3-7 atoms, said ring atom being bound to R ⁷ and R ⁸ . In addition to nitrogen, it comprises a carbocyclic atom, which carbocyclic atom is optionally substituted by one or more further heteroatoms, said cyclizing atom optionally having one or two or more It may be substituted by a group consisting of lower alkyl, ═O, or OR ⁹ .

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁸ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁸ is lower alkyl.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁹ is H.

According to another embodiment, the present invention relates to a compound of formula I wherein R ⁹ is lower alkyl.

の化合物
（式中、
Ｒ¹は、ＯＲ³で置換された低級アルキルであり；
Ｒ²は、Ｈ、又は、１つのＯＲ⁵基又は１つのＮＲ⁵Ｒ⁶基で置換された低級アルキルであり；
Ｒ³は、ハロゲン又はＯＲ⁷で置換されたアリールであるか、又は複素環に縮合したアリールであり；
Ｒ⁵及びＲ⁶は独立して、Ｈ、又は低級アルキルであり、或いは、基ＮＲ⁵Ｒ⁶は独立して、総原子数３〜６の環を形成していてもよく、前記環原子は、Ｒ⁵及びＲ⁶が結合する窒素に加えて、炭素環原子を含んでなり、前記炭素環原子は任意に、Ｎ又はＯから選択された１つの更なるヘテロ原子によって置換されていてもよく、前記環原子は任意に、ＯＲ⁷によって置換されていてもよく；
Ｒ⁷は、Ｈ、又は低級アルキルである）、又は医薬的に許容可能なその塩又はエステルに関する。 According to another embodiment, the present invention provides compounds of formula I

A compound of the formula
R ¹ is lower alkyl substituted with OR ³ ;
R ² is H or lower alkyl substituted with one OR ⁵ group or one NR ⁵ R ⁶ group;
R ³ is aryl substituted with halogen or OR ⁷ , or aryl fused to a heterocycle;
R ⁵ and R ⁶ are independently H or lower alkyl, or the group NR ⁵ R ⁶ may independently form a ring having a total number of 3 to 6, wherein the ring atom is In addition to the nitrogen to which R ⁵ and R ⁶ are bonded, it comprises a carbocyclic atom, said carbocyclic atom optionally being substituted by one further heteroatom selected from N or O The ring atom may optionally be substituted by OR ⁷ ;
R ⁷ is H or lower alkyl), or a pharmaceutically acceptable salt or ester thereof.

  The following compounds are examples of preferred embodiments according to the present invention.

4-amino-3- (4-bromo-2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide (Example 1),
4-amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide (Example 2),
4-Amino-3- (benzo [1,3] dioxol-5-yloxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide (Example 3) ,
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide (Examples 4a and 4b),
4-amino-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide (Example 5),
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1-methyl-ethyl) -amide (Example 6),
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-propyl) -amide (Example 7),
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2,3-dihydroxy-propyl) -amide (Example 8),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1,1-dimethyl-ethyl) -amide (Example 9),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1-hydroxymethyl-ethyl) -amide (Example 10),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-diethylamino-ethyl) -amide (Example 11),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide (Example 12a),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide hydrochloride (Example 12b),
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide methanesulfonate (Example 12c) ),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholin-4-yl-ethyl) -amide (Example 13a),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholin-4-yl-ethyl) -amide hydrochloride (Example 13b),
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (3-dimethylamino-2,2-dimethyl-propyl) -amide (Example 14) ,
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (5-diethylamino-1-methyl-pentyl) -amide (Example 15),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid amide (Example 16),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -amide (implementation) Example 17),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (3-methoxy-pyrrolidin-1-yl) -butyl] -amide (implementation) Example 18),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-piperidin-1-yl-butyl) -amide (Example 19),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-piperidin-1-yl-ethoxy) -ethyl] -amide (implementation) Example 20),
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (3-methoxy-piperidin-1-yl) -butyl] -amide Example 21),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-morpholin-4-yl-butyl) -amide (Example 22),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-morpholin-4-yl-ethoxy) -ethyl] -amide (implementation) Example 23),
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (4-methoxy-piperidin-1-yl) -butyl] -amide Example 24),
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [3- (2,3-dihydroxy-propoxy) -propyl] -amide (Example 25) ).

  The compounds of the present invention are selective for FGFR and / or KDR kinase. These compounds are useful in the treatment or control of cancer, particularly in the treatment or control of solid tumors, particularly breast tumors, lung tumors, colon tumors, or prostate tumors. These compounds are highly permeable to cell membranes and thus have an advantageous bioavailability profile, for example an improved oral bioavailability.

  The compounds of the present invention can be prepared by any conventional means. Suitable processes for synthesizing these compounds are described in the examples. In general, compounds of formula I can be prepared according to the following synthetic routes.

Scheme 1

Scheme 2

Separation of a mixture of stereoisomers into an optically pure stereoisomer (when the compound of formula I is chiral)
Isomeric structures of formula I may be separated, which can be carried out by well-known methods such as separation methods or chiral high pressure liquid chromatography (also known as chiral HPLC). Separation methods are well known and are summarized in "Enantiomers, Racemates, and Resolutions" (Jacques, J et al., John Wiley and Sons, NY, 1981). Chiral HPLC methods are also well known and are described in "Separation of Enantiomers by Liquid Chromatographic Methods" (Pirkle, WH and Finn, J., "Asymmetric Synthesis" Vol.1, Morrison, JD, Ed., Academic Press, Inc., NY 1983 , pp.87-124).

Conversion of a compound of formula I containing basic nitrogen to a pharmaceutically acceptable acid addition salt Conversion of a compound of formula I containing basic nitrogen to a pharmaceutically acceptable acid addition salt may be carried out. This can be done by conventional means. For example, the compound may be an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or a suitable organic material such as acetic acid, citric acid, tartaric acid, methanesulfonic acid, or p-toluenesulfonic acid. What is necessary is just to process with an acid.

Conversion of a compound of formula I containing a carboxylic acid group to a pharmaceutically acceptable alkali metal salt Conversion of a compound of formula I containing a carboxylic acid group to a pharmaceutically acceptable alkali metal salt may be performed. This can be done by conventional means. For example, the compound can be treated with an inorganic base such as lithium hydroxide, sodium hydroxide, or potassium hydroxide.

Conversion of compounds of formula I containing carboxylic acid groups or hydroxy groups into pharmaceutically acceptable esters Conversion of compounds of formula I containing carboxylic acid groups or hydroxy groups into pharmaceutically acceptable esters This can be done by conventional means. The conditions for forming the ester depend on the stability of the other functional groups in the molecule to the reaction conditions. If the rest of the molecule is stable to acidic conditions, the ester can be conveniently prepared by heating in an alcoholic solution of mineral acid (eg sulfuric acid). Other methods for preparing the ester are convenient when the molecule is not stable to acidic conditions, but in the presence of a coupling agent and optionally a further accelerator that can accelerate the reaction. And a method of treating a compound with an alcohol in the presence of an agent. Many such coupling agents are known to those skilled in the art of organic chemistry, and two examples include dicyclohexylcarbodiimide and triphenylphosphine / diethylazodicarboxylate. When dicyclohexylcarbodiimide is used as the coupling agent, the reaction is carried out at a temperature between about 0 degrees and about room temperature, preferably at about room temperature, in an inert solvent such as a halogenated hydrocarbon (eg dichloromethane). Is conveniently carried out by treating with any presence of alcohol, dicyclohexylcarbodiimide, and a catalytic amount (0-10 mol%) of N, N-dimethylaminopyridine. When triphenylphosphine / diethylazodicarboxylate is used as the coupling agent, the reaction is carried out at a temperature between about 0 degrees and about room temperature, preferably at about 0 degrees, such as an inert solvent such as ether (eg tetrahydrofuran). ) Or an aromatic hydrocarbon (for example, toluene), which is conveniently carried out by treating the acid with alcohol, triphenylphosphine, diethyl azodicarboxylate.

  According to another embodiment, the present invention comprises at least one compound of formula I, or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable excipient and / or carrier. A pharmaceutical composition comprising:

  These pharmaceutical compositions can be administered orally (eg in the form of tablets, coated tablets, dragees, hard or soft gelatin capsules, solutions, emulsions or suspensions). These compositions can also be administered rectally (eg in the form of suppositories) or parenterally (eg in the form of injection solutions).

  The pharmaceutical compositions of the present invention comprising a compound of formula I and / or a salt or ester thereof can be prepared by techniques known to those skilled in the art. Examples include the conventional mixing method, encapsulation method, dissolution method, granulation method, emulsification method, encapsulation method, sugar-coated tablet formation, or freeze-drying method. These pharmaceutical preparations may be formulated with therapeutically inert inorganic or organic carriers. Lactose, corn starch or derivatives thereof, stearic acid or its salts can be used as such carriers for tablets, coated tablets, dragees, and hard gelatin capsules. Suitable carriers for soft gelatin capsules include vegetable oils, waxes and fats. In the case of soft gelatin capsules, carriers are usually unnecessary depending on the nature of the active substance. Suitable carriers for the production of solutions and syrups include water, polyols, saccharose, invert sugar and glucose. Suitable carriers for injection include water, alcohols, polyols, glycerin, vegetable oils, phospholipids, and surfactants. Suitable carriers for suppositories include natural or hardened oils, waxes, fats and semi-liquid polyols.

  Pharmaceutical formulations contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavoring agents, osmotic pressure adjusting salts, buffers or coating agents, or antioxidants. It may be. The pharmaceutical formulation may also contain other therapeutically beneficial substances, including further active ingredients other than the active ingredient of formula I.

  As mentioned above, the compounds of the invention comprising compounds of formula I are useful for the treatment or control of cell proliferative disorders, such as prevention of new blood vessel formation (anti-angiogenesis) in solid tumors. These compounds and formulations containing said compounds are particularly useful for the treatment or control of solid tumors such as breast tumors, colon tumors, lung tumors, and prostate tumors.

  A therapeutically effective amount of a compound according to the invention means an amount of the compound that is effective to prevent, reduce or ameliorate symptoms of the disease in the patient being treated or prolong the life of the patient. Determination of a therapeutically effective amount is within the skill of those in the art.

  The therapeutically effective amount or dose of the compounds according to the invention can be selected within a wide range and can be determined by techniques known to those skilled in the art. Such dose is adjusted in combination with individual requirements in each specific case. Such requirements include the particular compound being administered, the route of administration, the condition being treated, and the patient being treated. In general, for oral or parenteral administration to an adult weighing about 70 Kg, a daily dose of about 10 mg to about 10,000 mg, preferably about 200 mg to about 1,000 mg is appropriate, if indicated May exceed the upper limit. The daily dose may be administered as a single dose or divided into several doses. In the case of parenteral administration, the dose may be administered by continuous infusion.

  The compounds of the invention may be used in combination (administered in combination or sequentially) with known anti-cancer treatments such as radiotherapy, or cytostatic or cytotoxic agents such as DNA interacting agents (eg Cisplatin or doxorubicin); topoisomerase II inhibitors (eg etoposide); topoisomerase I inhibitors (eg CPT-11 or topotecan); tubulin interactors (eg paclitaxel, docetaxel or epothilone); hormone agents (eg tamoxifen); thymidylate It may be used in combination with a synthetic inhibitor (eg 5-fluorouracil); and an antimetabolite (eg methotrexate). The compounds of formula I may also be useful in combination with modulators of p53 transactivation.

  When the combination product is formulated as a fixed dose, the product contains a compound of the invention within the above dose range and other pharmaceutically active substances or therapeutic agents within the approved dose range. For example, the early cdk1 inhibitor oromsin is known to synergize with well-known cytotoxic agents that induce apoptosis (J. Cell Sci. 1995, 108, 2897-2904). Compounds of formula I may be administered in tandem with such anti-cancer or cytotoxic agents where co-administration or combination with known anti-cancer or cytotoxic agents is inappropriate. The present invention is not limited as to the order of administration. That is, the compound of formula I may be administered before or after administration of a known anticancer agent or cytotoxic agent. For example, the cytotoxic activity of the cdk inhibitor flavopiridol is affected by the order of administration with anticancer agents (Cancer Research, 1997, 57, 3375).

  The invention also relates to the following novel intermediates useful for the synthesis of compounds of formula I:

3-methyl-5H-thieno [3,2-c] pyridin-4-one (intermediate 4),
7-iodo-3-methyl-5H-thieno [3,2-c] pyridin-4-one (Intermediate 5),
3-methyl-4-oxo-4,5-dihydro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 6),
4-chloro-3-methyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 7),
3-bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 8),
3- (4-bromo-2,6-difluoro-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 9),
4-amino-3- (4-bromo-2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 10),
4-Amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (Intermediate 11)
3- (4-bromo-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 12),
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 13),
4-chloro-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 14),
4-amino-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (intermediate 15), and 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3 , 2-c] pyridine-7-carboxylic acid (intermediate 16).

  The following examples illustrate preferred methods for synthesizing the compounds and formulations of the present invention.

Intermediate 1
4-methyl-2-thiophenecarboxaldehyde

  A solution of 3-methylthiophene (58.90 g, 0.60 mol) (Fluka) in anhydrous ether (600 mL) was stirred and cooled in an ice-water bath. This solution was treated dropwise with n-butyllithium in pentane (2M, 450 mL, 0.90 mol) (Aldrich) for 15 minutes. After stirring for 2 hours at room temperature, the mixture was cooled in an ice-water bath and treated dropwise with N, N-dimethylformamide (48.24 g, 0.66 mol) (Fisher) for 5 minutes, followed by overnight at room temperature. Stir. The mixture was diluted with ether (600 mL) and washed with water and brine. After drying (sodium sulfate), ether was filtered off and evaporated on a rotary evaporator without vacuum, yielding 114 g of red liquid. The liquid was purified by chromatography on a pad of silica gel 60 (1 Kg, 70-230 mesh) eluting with 40% dichloromethane-hexane. Evaporation without vacuum gave a mixture of 4-methyl-2-thiophenecarboxaldehyde and 3-methyl-2-thiophenecarboxaldehyde (approximately 5: 1) as a light red oil (yield 56. 62g, 74.7%).

Intermediate 2
3- (4-Methyl-thiophen-2-yl) -acrylic acid

  4-Methyl-2-thiophenecarboxaldehyde (56.62 g, 0.448 mol) in pyridine (550 mL) (from Intermediate 1 above, containing 3-methyl-2-thiophenecarboxaldehyde), malonic acid (186.77 g A solution of 1.79 mol) (Aldrich) and piperidine (1.90 g, 0.022 mol) (Fluka) was heated at reflux with stirring overnight. The reaction mixture was evaporated to dryness. The resulting residue was dissolved in dichloromethane and washed successively with 3N hydrochloric acid, water, and brine. The organic phase was dried (sodium sulfate), filtered and evaporated to give 3- (4-methyl-thiophen-2-yl) -acrylic acid as a tan solid (yield 49.52 g, 65 .7%).

Intermediate 3
3- (4-Methyl-thiophen-2-yl) -acryloyl azide

  3- (4-Methyl-thiophen-2-yl) -acrylic acid (49.52 g, 0.294 mol) (from Intermediate 2 above) and triethylamine (44.68 g, 0.441 mol) in acetone (2000 mL) ( To a solution of (Aldrich), ethyl chloroformate (35.14 g, 0.323 mol) (Aldrich) was added while stirring and cooling in an ice-water bath. After stirring for 20 minutes at room temperature, sodium azide (28.70 g, 0.441 mol) (Aldrich) was added and stirred continuously for another 20 minutes at room temperature. The acetone was then evaporated under reduced pressure and the residue was diluted with water. This was extracted with dichloromethane. The organic extract was washed with brine, dried (sodium sulfate), filtered and concentrated to give 3- (4-methyl-thiophen-2-yl) -acryloyl azide as a brown solid (yield 48). .51 g, 85.4%).

Intermediate 4
3-Methyl-5H-thieno [3,2-c] pyridin-4-one

  Method A: A mixture of 3- (4-methyl-thiophen-2-yl) -acryloyl azide (69.21 g, 0.358 mol) (from Intermediate 3 above) and xylene (700 mL) was stirred and 0.5 Heated at reflux for hours. Iodine (0.45 g, 1.79 mmol) was added and the mixture was heated at reflux overnight. The reaction mixture was cooled and stirred with aqueous sodium bisulfite solution for 5 minutes. The suspension was filtered, washed with ether and dried by suction to give 3-methyl-5H-thieno [3,2-c] pyridin-4-one as a brown solid (yield 31. 28g, 52.8%).

  Method B: 3- (4-Methyl-thiophen-2-yl) -acryloyl azide (1.54 g; 7.95 mol) (from Intermediate 3 above) was dissolved in meta-xylene (16 mL). The solution was heated in an oil bath at 105-115 ° C. for 30 minutes until the end of nitrogen evolution. A small number of iodine crystals were then added to the reaction and the oil bath temperature was raised to 145-150 ° C. The reaction was heated at reflux for 6 hours. Upon cooling, solids precipitated from the solution. Filtration and drying yielded 3-methyl-5H-thieno [3,2-c] pyridin-4-one (yield 1.05 g; 80.1%).

_{C 8 H 7 NOS [(M} +)] for HRMS (EI +) m / z calcd: 165.0248. Found: 165.0250.

Intermediate 5
7-Iodo-3-methyl-5H-thieno [3,2-c] pyridin-4-one

3-Methyl-5H-thieno [3,2-c] pyridin-4-one (24.27 g, 0.146 mol) (from Intermediate 4 above) and N-iodo in N, N-dimethylformamide (1000 mL) A solution of succinimide (34.70 g, 0.154 mol) (Avocado) was stirred overnight at room temperature. The reaction mixture was concentrated under reduced pressure and the residue was stirred with ether (1000 mL) for 0.5 h. The suspension was filtered, washed with ether and dried by suction to give 7-iodo-3-methyl-5H-thieno [3,2-c] pyridin-4-one as a brown solid. (Yield 41.88 g, 97.9%).
_{C 8 H 6 INOS [(M} +)] for HRMS (EI +) m / z calcd: 290.9215. Found: 290.9210.

Intermediate 6
3-Methyl-4-oxo-4,5-dihydro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  7-Iodo-3-methyl-5H-thieno [3,2-c] pyridin-4-one (1.14 g, 3.29 mmol) (from intermediate 5 above), triethylamine (2. A stirred suspension of 5 mL, 17.94 mmol) (Aldrich), and bis (triphenylphosphine) palladium (II) chloride (0.14 g, 0.2 mmol) (Aldrich) was degassed with argon, Saturated with carbon oxide. The mixture was stirred overnight with heating in a 75 ° C. oil bath under an atmospheric pressure carbon monoxide seal. After cooling, the reaction mixture was concentrated under reduced pressure to remove a portion of ethanol (about 20%). The solid formed is collected by filtration, washed with ethanol-diethyl ether (1: 1), then diethyl ether and finally dried under vacuum to give 3-methyl-4-oxo-4,5 -Dihydro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester was obtained (yield 0.78 g, 84.0%).

_{C 11 H 11 NO 3 S [} (M +)] for HRMS (EI +) m / z calcd: 237.0460. Found: 237.0451.

Intermediate 7
4-Chloro-3-methyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  3-Methyl-4-oxo-4,5-dihydro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (2.43 g, 10.24 mmol) (from intermediate 6 above) and N, N -Diisopropylethylamine (2.4 mL, 13.87 mmol) (Fluka) was stirred with cooling in an ice-water bath. The mixture was treated gently with phosphorus oxychloride (7.8 mL, 83.68 mmol) (Fluka) and then allowed to recover to room temperature. N, N-dimethylformamide (1.0 mL, 12.86 mmol) was then added and the mixture was stirred for 30 minutes with heating at 70 ° C. Additional N, N-dimethylformamide (0.5 mL, 6.43 mmol) was added and the mixture was heated at 70 ° C. for an additional 30 minutes. After cooling, ice was added to the solution and the mixture was extracted with ethyl acetate. The organic extract was washed with water, saturated aqueous sodium bicarbonate, water and brine. The aqueous phase was backwashed with ethyl acetate. The combined ethyl acetate solution was dried (sodium sulfate), filtered and concentrated under reduced pressure. The residue is purified by flash chromatography on silica gel (Biotage 65M, 5:95 ethyl acetate: hexanes) to give 4-chloro-3-methyl-thieno [3,2-c] pyridine-7-carboxylic acid. The ethyl ester was obtained (yield 1.57 g, 60.0%).

_{C 11 H 10 ClNO 2 S [} (M +)] for HRMS (EI +) m / z calcd: 255.0121. Found: 255.0119.

Intermediate 8
3-Bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  Solution of 4-chloro-3-methyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (0.81 g, 3.17 mmol) (from Intermediate 7 above) in carbon tetrachloride (50 mL) N-bromosuccinimide (0.73 g, 4.12 mmol) (Avacado) and 2,2′-azobisisobutyronitrile (52 mg, 0.32 mmol) (Aldrich) were added respectively. The reaction mixture was heated at 80 ° C. for 24 hours. The mixture was then cooled and concentrated under reduced pressure. The residue was purified by chromatography (ethyl ether-hexane, 1: 4, V / V) to give 3-bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester. Obtained as a white solid (yield 0.7 g, 66%).

_{C 11 H 9 BrClNO 2 S [} (M +)] for HRMS (EI +) m / z calcd: 332.9226. Found: 332.9224.

Intermediate 9
3- (4-Bromo-2,6-difluoro-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  3-Bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (265 mg, 0. 1) in a mixture of tetrahydrofuran and N, N-dimethylformamide (10 mL, 5: 1). 79 mmol) (from Intermediate 8 above) and 2,6-difluoro-4-bromo-phenol (166 mg, 0.79 mmol) (Alfa) were treated with potassium carbonate (110 mg, 0.79 mmol). After stirring at room temperature for 15 minutes, the reaction mixture was heated to 65 ° C. and stirred at that temperature for an additional 5.5 hours. The mixture was then cooled and partitioned between dichloromethane and water. The organic phase is dried over sodium sulfate, filtered and concentrated, and the residue is purified by chromatography on a silica gel column and 0-30% diethyl ether in hexane to give the product. It was. 3- (4-Bromo-2,6-difluoro-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester by precipitation from chloroform with excess hexane Was obtained as a white solid (yield 270 mg, 73%).

_{C 17 H 11 BrClF 2 NO 3} S + H [M + H] HRMS m / z calculated for ^+: 461.9373. Found: 461.9377.

Intermediate 10
4-Amino-3- (4-bromo-2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  3- (4-Bromo-2,6-difluoro-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (200 mg, 0.43 mmol) in dioxane (10 mL) Ammonia gas was bubbled into the solution (from intermediate 9 above) in a pressure reactor for 5 minutes. The reaction mixture was sealed and stirred at 130 ° C. for 9 hours and then at room temperature overnight. The solution was then evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography using 0-100% gradient ethyl acetate in hexane and precipitation from tetrahydrofuran with excess hexane to give 4-amino-3- (4-Bromo-2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester was obtained as a white solid (yield 130 mg, 67%).

_{C 17 H 13 BrF 2 N 2} O 3 S HRMS m / z calculated for [M ^+]: 441.9798. Found: 441.9786.

Intermediate 11
4-Amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  3-Bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (400 mg, 1.19 mmol) in a mixture of tetrahydrofuran (8 mL) and dichloromethane (2 mL) (intermediate above) 8) was added 2-chloro-4-methoxyphenol (192 mg, 1.21 mmol) (Aldrich) followed by potassium carbonate (167 mg, 1.21 mmol). Once the starting material was consumed as confirmed by thin layer chromatography, the reaction mixture was partitioned between dichloromethane and water. The organic phase was dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography using a 0-30% gradient of diethyl ether in hexane to give the intermediate 4-chloro-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [ 3,2-c] pyridine-7-carboxylic acid ethyl ester was obtained.

  In a solution of this intermediate 4-chloro-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester in dioxane in a pressure reactor. For 5 minutes at room temperature. The reaction vessel was then sealed and the mixture was stirred for 12 hours at 120 ° C. and for 48 hours at room temperature. The reaction mixture was then evaporated under reduced pressure. The residue was purified by chromatography on a Biotage system using a 20-40% gradient of ethyl acetate in hexane to give 4-amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3 , 2-c] pyridine-7-carboxylic acid ethyl ester was obtained as a white powder (yield 80 mg, 17%).

_{C 18 H 17 ClN 2 O 4} S HRMS m / z calculated for [M ^+]: 392.0598. Found: 392.0582.

Intermediate 12
3- (4-Bromo-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  Of potassium carbonate (0.67 g, 4.85 mmol) and 4-bromophenol (0.78 g, 4.47 mmol) (Aldrich) in a mixture of tetrahydrofuran and N, N-dimethylformamide (5: 1, 40 mL). The suspension was heated at 65-70 ° C. for 3 hours. 3-Bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (1.41 g, 4.21 mmol) (from intermediate 8 above) was added and another tetrahydrofuran and N, Rinse with a mixed solvent with N-dimethylformamide (5: 1, 13 mL). Heating was continued for 20 hours. The reaction mixture was cooled and concentrated under reduced pressure. The residue was partitioned between dichloromethane and water. The organic phase was washed with water and brine, dried (sodium sulfate), filtered and concentrated. The crude product was crystallized from hot acetonitrile to give 3- (4-bromo-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (yield: 1 .15 g, 66.1%).

_{C 17 H 13 BrClNO 3 S +} H [(M + H) +] for HRMS (ES +) m / z calcd: 425.9561. Found: 425.9562.

Intermediate 13
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  Method A: A solution of ammonia in dioxane (0.5N, 200 mL, 100 mmol) (Aldrich) in a pressure tube contained 3- (4-bromo-phenoxymethyl) -4-chloro-thieno [3,2-c]. Pyridine-7-carboxylic acid ethyl ester (2.1 g, 4.9 mmol) (from Intermediate 12 above) was added. The reaction mixture was sealed under nitrogen (50 psi) and heated at 100 ° C. for 48 hours. The mixture was cooled and concentrated under reduced pressure. The residue is purified by chromatography (ethyl acetate: hexane, 1: 1, then ethyl acetate) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine- 7-carboxylic acid ethyl ester was obtained as a white solid (yield: 1.5 g, 75%).

  Method B: 4-Chloro-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (0.95 g; 2.05 mmol) in dry dioxane (21 mL) in a pressure bottle (above Ammonia gas was bubbled into the solution of intermediate 12) for 15 minutes. The bottle was then capped and the solution was heated at 120-150 ° C. The reaction was monitored by liquid chromatography analysis and after 15 hours it was recharged with ammonia. The reaction was stopped after 40 hours. The reaction mixture was concentrated. The residue was partitioned between dichloromethane and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The crude product was purified by flash chromatography (Biotage 40M; ethyl acetate-hexane gradient (10-50% ethyl acetate) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2- c] Pyridine-7-carboxylic acid ethyl ester was obtained (yield: 0.65 g, 76.32%).

_{C 17 H 15 BrN 2 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 407.0060. Found: 407.0060.

Intermediate 14
4-Chloro-3-phenoxymethyl-thieno [3,2-c] pyridine-7-
Carboxylic acid ethyl ester

  A suspension of potassium carbonate (31 mg, 0.22 mmol) and phenol (22 mg, 0.23 mmol) in a mixture with tetrahydrofuran-dimethylformamide (2.8 mL, 5: 1) was heated at 65 ° C. for 2 hours. . 3-Bromomethyl-3-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (75 mg, 0.22 mmol) (from Intermediate 12 above) was added and heating was continued overnight. The reaction mixture was cooled and concentrated. The residue was partitioned between dichloromethane and water. The organic phase was washed with brine (2X), dried over sodium sulfate, filtered and concentrated. The crude product is purified by flash chromatography (Biotage 40S: 75: 25 dichloromethane-hexane) to give 4-chloro-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester. (Yield 19.4 mg, 24.9%).

Intermediate 15
4-Amino-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester

  4-chloro-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (38 mg; 0.11 mmol) in dioxane (2.4 mL) in a pressure bottle (from intermediate 14 above) ) Was bubbled into the solution for 30 minutes. The bottle was then capped and the clear colorless solution was heated in an oil bath at 115-125 ° C. overnight. The crude orange mixture was concentrated and purified by flash chromatography (Biotage 12M; ethyl acetate-hexane gradient (15-100% ethyl acetate) to give 4-amino-3-phenoxymethyl-thieno [3,2-c Pyridine-7-carboxylic acid ethyl ester was obtained (yield: 14 mg, 39.1%).

  A substantial amount of unreacted 4-chloro-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (16 mg) was recovered from chromatography.

Intermediate 16
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-
7-carboxylic acid

  4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (0.75 g; 1) in tetrahydrofuran-methanol (13 mL, 3: 1). 84 mmol) (from intermediate 13 above) was added an aqueous solution of sodium hydroxide (1.0 N, 3.1 mL, 3.1 mmol) and the mixture was heated at 35-40 ° C. for 18 hours. The crude reaction mixture was concentrated and azeotroped with toluene. The solid residue was triturated with ethyl acetate. The solid was then suspended in water and treated with dilute hydrochloric acid (1.0 N, 3.4 mL). After stirring for 30 minutes, the solid was collected, washed with water, then with diethyl ether, and dried to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c]. Pyridine-7-carboxylic acid was obtained (yield: 0.67 g, 95.5%).

_{C 15 H 11 BrN 2 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 378.9747. Found: 378.9747.

Intermediate 17
4- (4-Aminobutyl) morpholine

4-Bromobutylphthalimide (5.0 g, 17.7 mmol) (Lancaster), morpholine (2.0 mL, 23.0 mmol) (Aldrich), and triethylamine (5.0 mL, 35.9 mmol) in absolute ethanol (50 mL) The solution was heated at reflux for 16 hours. Ethanol was removed under reduced pressure. The residue was diluted with dichloromethane and washed with water and brine. After drying (MgSO ₄ ), dichloromethane was evaporated under reduced pressure. The residue was purified by flash chromatography eluting with 4% methanol in dichloromethane to give 2- (4-morpholin-4-yl-butyl) -isoindole-1,3-dione (yield 4. 44 g, 87%).

To a solution of 2- (4-morpholin-4-yl-butyl) -isoindole-1,3-dione (4.44 g, 15.4 mmol) in absolute ethanol (100 mL) was added hydrazine hydrate (2.0 mL). 41.2 mmol) (Aldrich) was added and the mixture was heated at reflux for 2 hours. The mixture was then cooled, filtered and the precipitate washed with absolute ethanol. The combined filtrate and washings were concentrated under reduced pressure. The resulting residue was suspended in dry tetrahydrofuran (100 mL) and cooled in ice. Benzyl chloroformate (Aldrich) (7.5 mL of 50% toluene solution, 52.5 mmol) was added dropwise and the mixture was stirred for 18 hours at room temperature. Excess reagent was quenched with methanol. The solvent was removed under reduced pressure. The residue was diluted with water and the resulting solution was acidified to pH 1 (with dilute hydrochloric acid). The aqueous solution was washed with dichloromethane, then treated with excess sodium carbonate (to pH 10) and extracted with ethyl acetate (3 × 100 mL). The combined ethyl acetate layers were dried (MgSO ₄ ) and filtered. The solvent is then removed under reduced pressure and the residue is purified by flash chromatography eluting with a gradient of 0-5% methanol in dichloromethane to give N- (benzyloxycarbonyl) -4- (4-aminobutyl). ) Obtained morpholine (yield 2.19 g, 49%).

  A solution of N- (benzyloxycarbonyl) -4- (4-aminobutyl) morpholine (2.19 g, 7.49 mmol) in methanol (50 mL) was added at 54 psi with 10% Pd / C (0.2 g). Hydrogenated over 18 hours. The mixture was filtered through a Celite® pad and concentrated under reduced pressure to give 4- (4-aminobutyl) morpholine (yield 1.43 g, 100%). This was used without further purification.

Intermediate 18
2- (2-Pyrrolidin-1-yl-ethoxy) -ethylamine

  To a solution of 2- (2-aminoethoxy) ethanol (3.5 g, 33.3 mmol) (Aldrich) in dichloromethane (50 mL) at 0 ° C. was added N-carboethoxyphthalimide (Aldrich) and triethylamine. The mixture was stirred for 1 day at room temperature and then concentrated under reduced pressure. The residue is then purified by flash chromatography eluting with ethyl acetate-hexane (2: 1, V / V) to give 2- [2- (2-hydroxy-ethoxy) -ethyl] -isoindole- 1,3-dione was obtained (yield 3.77 g, 48%).

  2- [2- (2-Hydroxy-ethoxy) -ethyl] -isoindole-1,3-dione (3.77 g, 16.03 mmol) and carbon tetrabromide (6) in dichloromethane (60 mL) at 0 ° C. To a solution of .38 g, 19.23 mmol) (Aldrich) was added triphenylphosphine (5.04 g, 19.23 mmol) (Aldrich). The mixture was stirred for 18 hours at room temperature. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with ethyl acetate-hexane (2: 1, V / V) to give 2- [2- (2-bromo-ethoxy ) -Ethyl] -isoindole-1,3-dione was obtained (yield 4.0 g, 84%).

2- [2- (2-Bromo-ethoxy) -ethyl] -isoindole-1,3-dione (4.0 g, 13.4 mmol), pyrrolidine (1.46 mL, 17.4 mmol) in absolute ethanol (70 mL) ) (Aldrich) and a solution of triethylamine (3.74 mL, 26.8 mmol) were heated at reflux for 18 hours. Ethanol was removed under reduced pressure. The residue was diluted with dichloromethane and washed with water and brine. After drying (MgSO ₄ ), dichloromethane was evaporated under reduced pressure. The residue was purified by flash chromatography eluting with a 5-10% methanol gradient in dichloromethane to give 2- [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -isoindole-1,3- Dione was obtained (yield 1.56 g, 40%).

To a solution of 2- [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -isoindole-1,3-dione (1.56 g, 5.41 mmol) in absolute ethanol (20 mL) was added hydrazine water. The Japanese product (1.0 mL, 20.6 mmol) (Aldrich) was added. The mixture was heated at reflux for 2 hours, cooled, filtered and the precipitate washed with absolute ethanol. The filtrate was concentrated and the residue was suspended in dry tetrahydrofuran (30 mL) and cooled in ice. Benzyl chloroformate (Aldrich) (2.62 mL of 50% toluene solution, 18.39 mmol) was added dropwise and the mixture was stirred for 18 hours at room temperature. Excess reagent was quenched with methanol and the solvent was removed under reduced pressure. The residue was diluted with water and the resulting solution was acidified to pH 1 (with dilute hydrochloric acid), washed with dichloromethane, then treated with excess sodium carbonate (to pH 10) and extracted with ethyl acetate (3 × 50 mL). . The combined ethyl acetate layers were dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography eluting with a gradient of 0-5% methanol in dichloromethane to give [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -carbamic acid benzyl ester. (Yield 1.2 g, 76%).

  A solution of [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -carbamic acid benzyl ester (1.2 g, 4.1 mmol) in methanol (50 mL) was added to 10% Pd / C (0.1 g ) At 50 psi for 18 hours. The mixture was filtered through a Celite® pad and concentrated under reduced pressure to give 2- (2-pyrrolidin-1-yl-ethoxy) -ethylamine (yield 0.85 g, 99%). This was used without further purification.

Intermediate 19
4- (4-Methoxy-piperidin-1-yl) -butylamine

  N-formyl-4-hydroxypiperidine was synthesized from 4-hydroxypiperidine (Aldrich) according to the literature procedure of Baker, W. R. et al., J. Med. Chem., 1992, 35, 1722-1734.

To a solution of N-formyl-4-hydroxypiperidine (10.0 g, 77.4 mmol) in tetrahydrofuran (100 mL) was added sodium hydride (3.41 g, 60% in oil, 85.2 mmol) at 0 ° C. (Aldrich) Followed by stirring at room temperature for 2 hours. The mixture was then re-cooled to 0 ° C. and iodomethane (5.3 mL, 85.2 mmol) (Aldrich) was added dropwise. The mixture was stirred for 18 hours at room temperature. The reaction was carefully quenched with water and extracted with ethyl acetate (3 × 50 mL). The combined ethyl acetate layers were dried (MgSO ₄ ) and filtered. The solvent was removed under reduced pressure and the residue was purified by flash chromatography eluting with 4% methanol in dichloromethane to give 4-methoxy-piperidine-1-carbaldehyde (yield 5.63 g). 51%).

A solution of 4-methoxy-piperidine-1-carbaldehyde (5.63 g, 39.30 mmol) and potassium hydroxide (7.37 g, 0.13 mol) in water (40 mL) was stirred at room temperature for 1 day. The reaction mixture was extracted with ether (4 × 20 mL) and the combined ethyl acetate layers were dried (MgSO ₄ ) and filtered. This was concentrated under reduced pressure to give 4-methoxy-piperidine which was used without further purification (yield 2.43 g, 33%).

4-Bromobutylphthalimide (5.0 g, 17.7 mmol) (Lancaster), 4-methoxy-piperidine (2.43 g, 21.3 mmol), and triethylamine (5.0 mL, 35.9 mmol) in absolute ethanol (50 mL). ) Was heated at reflux for 18 hours. Ethanol was removed under reduced pressure. The residue was diluted with dichloromethane and washed with water and brine. After drying (MgSO ₄ ) and filtration, the mixture was concentrated under reduced pressure. The residue was purified by flash chromatography eluting with 4% methanol in dichloromethane to give 2- [4- (4-methoxy-piperidin-1-yl) -butyl] -isoindole-1,3-dione. Obtained (yield 4.06 g, 73%).

To a solution of 2- [4- (4-methoxy-piperidin-1-yl) -butyl] -isoindole-1,3-dione (4.06 g, 12.87 mmol) in absolute ethanol (100 mL) was added hydrazine water. The Japanese product (2.0 mL, 41.2 mmol) (Aldrich) was added. The reaction mixture was heated at reflux for 2 hours, then cooled, filtered and the precipitate washed with absolute ethanol. The filtrate was concentrated under reduced pressure and the residue was suspended in dry tetrahydrofuran (100 mL) and cooled in ice. Benzyl chloroformate (Aldrich) (6.25 mL of 50% toluene solution, 43.77 mmol) was added dropwise followed by stirring at room temperature for 18 hours. Excess reagent was quenched with methanol and the solvent was removed under reduced pressure. The residue was diluted with water and the resulting solution was acidified to pH 1 (with dilute hydrochloric acid), washed with dichloromethane, then treated with excess sodium carbonate (to pH 10) and extracted with ethyl acetate (3 × 100 mL). . The combined ethyl acetate layers were dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography eluting with a 5-10% methanol gradient in dichloromethane to give [4- (4-methoxy-piperidin-1-yl) -butyl] -carbamic acid benzyl ester. (Yield 1.9 g, 46%).

  A solution of [4- (4-methoxy-piperidin-1-yl) -butyl] -carbamic acid benzyl ester (1.9 g, 5.93 mmol) in methanol (30 mL) was added to 10% Pd / C (0.19 g ) At 50 psi for 18 hours. The mixture was filtered through a Celite® pad and the filtrate was concentrated under reduced pressure to give 4- (4-methoxy-piperidin-1-yl) -butylamine (yield 1.43 g, 100%). This was used without further purification.

Intermediate 20
3- (2,2-Dimethyl- [1,3] dioxolan-4-yl-methoxy) -propylamine

2,2-Dimethyl-1,3-dioxolane-4-methanol (26.43 g, 0.20 mol) (Aldrich) and acetonitrile (26.33 mL, 0.40 mol) (Aldrich) in dry tetrahydrofuran (500 mL) at 0 ° C. ) Was added sodium hydride (1.6 g, 60% in oil, 40 mmol) (Aldrich). The reaction mixture was stirred at room temperature for 1 hour, then water (100 mL) was added dropwise and the resulting suspension was concentrated under reduced pressure. Water (200 mL) was added again and the mixture was extracted with dichloromethane (2 × 300 mL). The extracts were combined, dried (MgSO ₄ ), filtered and concentrated to give an oil. The oil was distilled under reduced pressure to give 3- (2,2-dimethyl- [1,3] dioxolan-4-yl-methoxy) -propionitrile (yield 26.07 g, 70%; Boiling point 86-105 ° C./0.5 mmHg).

To a solution of 3- (2,2-dimethyl- [1,3] dioxolan-4-yl-methoxy) -propionitrile (13.89 g, 75.0 mmol) in methanol (450 mL) was added cobalt (II) chloride. (19.48 g, 0.15 mol) (Aldrich) was added. To this stirred and cooled (ice water bath) solution was added sodium borohydride (28.37 g, 0.75 mol) (Aldrich). Stirring was continued for 1 hour and then concentrated aqueous ammonium hydroxide (250 mL) was added. The resulting suspension was filtered and concentrated under reduced pressure to remove methanol. The mixture was extracted with dichloromethane (2 × 300 mL) and the combined extracts were dried (MgSO ₄ ) and concentrated to give an oil. The oil was distilled under reduced pressure to give 3- (2,2-dimethyl- [1,3] dioxolan-4-yl-methoxy) -propylamine (yield 7.95 g, 56%; boiling point). 75-82 ° C./0.6 mmHg).

Example 1
4-Amino-3- (4-bromo-2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide

  4-amino-3- (4-bromo-2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester in ethanolamine (approximately 2 mL) (Aldrich) A solution of 51 mg, 0.11 mmol) (from Intermediate 10 above) was stirred at 75 ° C. for 8 hours and at room temperature overnight. The mixture was then partitioned between ethyl acetate and water. The organic phase was dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography using a 0-30% methanol gradient in dichloromethane followed by precipitation from tetrahydrofuran with excess hexane to give 4-amino-3- (4-bromo -2,6-Difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide was obtained as a white solid (yield 22 mg, 42%).

_{C 17 H 14 BrF 2 N 3} O 3 S + H [M + H] HRMS m / z calculated for ^+: 457.9980. Found: 457.9984.

Example 2
4-Amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide

  4-amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (80 mg, 0.20 mmol) (from intermediate 11 above). , Dissolved in a mixture of ethanolamine (2 mL) (Aldrich) and dimethyl sulfoxide (approximately 1 mL). The mixture was stirred at 80 ° C. overnight and cooled to room temperature. Water was added and the mixture was filtered to collect the white precipitate that formed. The precipitate was purified by chromatography on a silica gel column with a 0-10% methanol gradient in dichloromethane. The pure fractions were combined and concentrated, and the residue was precipitated from dimethyl sulfoxide with excess water to give 4-amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2 -C] Pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide was obtained as a white powder (yield 20 mg, 25%).

_{C 18 H 18 ClN 3 O 4} S HRMS m / z calculated for [M ^+]: 407.0707. Found: 407.0700. KDR IC ₅₀ 0.5089 μM, FGFR IC ₅₀ 2.559 μM.

Example 3
4-Amino-3- (benzo [1,3] dioxol-5-yloxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide

  3-Bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (400 mg, 1.19 mmol) (from intermediate 8 above) in tetrahydrofuran (8 mL) and dichloromethane (2 mL) Was treated with sesamol (167 mg, 1.01 mmol) (Aldrich) and potassium carbonate (167 mg, 1.21 mmol) and stirred at room temperature until the starting material was confirmed by thin layer chromatography. . The reaction mixture was partitioned between dichloromethane and water. The organic phase was dried over sodium sulfate, filtered and concentrated. The residue 3- (benzo [1,3] dioxol-5-yloxymethyl) -4-chloro was purified by silica gel column chromatography with a 0-30% diethyl ether gradient in hexane. -Thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester was obtained.

  In a solution of intermediate 3- (benzo [1,3] dioxol-5-yloxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester dissolved in dioxane Ammonia gas was bubbled at room temperature for 5 minutes in a pressure tube. The reaction vessel was then sealed and the mixture was stirred at 120 ° C. for 12 hours and then at room temperature for 48 hours. The solvent was removed by evaporation under reduced pressure. The resulting residue was purified by flash chromatography (Biotage system with an ethyl acetate gradient in hexane of 20-40%) to give 4-amino-3- (benzo [1,3] dioxol-5-yloxy. Methyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester was obtained. This was used in the next step without further characterization.

  This 4-amino-3- (benzo [1,3] dioxol-5-yloxymethyl) -thieno [3,2-c] pyridine-7 in ethanolamine (2 mL) (Aldrich) and dimethyl sulfoxide (1 mL) The carboxylic acid ethyl ester solution was heated in a 120 ° C. oil bath and stirred until the starting material was consumed as confirmed by thin layer chromatography. The reaction mixture was then cooled and treated with water. The formed precipitate was collected by filtration, washed with water and dried to give 4-amino-3- (benzo [1,3] dioxol-5-yloxymethyl) -thieno [3,2- c] Pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide was obtained as a white powder (yield 15 mg, 3%).

_{C 18 H 17 N 3 O 5} S HRMS m / z calculated for [M ^+]: 387.0889. Found: 387.0888.

Example 4a
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide

  4-amino-3- (4-bromophenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (30 mg, 0.074 mmol) in dimethyl sulfoxide (0.5 mL) (intermediate above) 13) and ethanolamine (0.50 mL, 8.31 mmol) (Aldrich) were heated in a 70 ° C. oil bath for 10 hours. The reaction was diluted with ethyl acetate and washed with water. The organic phase was concentrated and the residue was treated with water. The formed precipitate was collected but confirmed to still contain starting material. This solid and mother liquor are combined with additional 4-amino-3- (4-bromophenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (14 mg; 0.034 mmol), Dissolved in dimethyl sulfoxide (0.5 mL). Ethanolamine (1.0 mL, 16.62 mmol) was added and the mixture was heated at 75 ° C. overnight. The crude reaction was diluted with water, resulting in the precipitation of a milky white solid. Ethyl acetate was added but the solid did not dissolve. The solid was collected and dried to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide. (Yield 31.5 mg (90% purity); 62.10%).

  A portion of the above material (22 mg) was dissolved in dimethyl sulfoxide (0.5 mL) and re-treated with ethanolamine (1.0 mL, 16.62 mmol) at 75 ° C. for 24 hours. The crude reaction was diluted with water and then with ethyl acetate to precipitate a solid. The solid was collected and dried to give pure 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide. (Yield 20 mg, 43.8% overall).

_{C 17 H 16 BrN 3 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 422.0169. Found: 422.0173. _{KDR IC 50 0.0200μM, FGFR IC 50} 0.0724μM, VEGF-HUVEC 0.264μM, FGF-HUVEC 2.762μM.

Example 4b
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amidobis-methanesulfonate

  4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide (0.05 g, 0) in methanol (5 mL). .12 mmol) (from Example 4a above) was treated with metasulfonic acid (7.7 μL, 0.12 mmol). The mixture was stirred for 2 days at room temperature and then concentrated under reduced pressure. The residue was suspended in water. The solid was filtered and dried to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amidobis- Methanesulfonate was obtained as a white powder (Yield: 25 mg, 34%).

Example 5
4-Amino-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide

  Solution of 4-amino-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (31 mg, 0.094 mmol) (from Intermediate 15 above) in dimethyl sulfoxide (0.5 mL) Was treated with ethanolamine (1.0 mg, 16.62 mmol) (Aldrich) in a pressure bottle and heated at 75 ° C. for 16 hours. When the crude reaction mixture was diluted with water, a solid precipitated. The solid was collected and confirmed to still contain 15% unreacted starting material. This solid was recombined with the mother liquor and further retreated with ethanolamine (1.0 mL) at 75 ° C. for 19 hours. The crude reaction mixture was diluted with ethyl acetate and water. The resulting solid was collected, washed with water and diethyl ether, and then triturated with acetonitrile to give 4-amino-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ( 2-Hydroxy-ethyl) -amide was obtained (Yield: 16.7 mg, 51.5%).

_{C 17 H 17 N 3 O 3} S + H [(M + H) +] for HRMS (+ ES) m / z calcd: 344.1064. Found: 344.1066.

Example 6
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1-methyl-ethyl) -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (50 mg, 0.13 mmol) (from intermediate 13 above) and racemic 2- A mixture of amino-1-propanol (2.6 g, 35 mmol) (Aldrich) was heated at 150 ° C. for 6 hours. The mixture was cooled, diluted with ethyl acetate (50 mL) and then washed with water. The aqueous phase was extracted with ethyl acetate (50 mL). The organic phases were separated and combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by chromatography (ethyl acetate-methanol, 85:15) to give rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7- The carboxylic acid (2-hydroxy-1-methyl-ethyl) -amide was obtained as a white solid (yield 30 mg, 55%).

_{C 18 H 18 BrN 3 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 436.0325. Found: 436.0329.

Example 7
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-propyl) -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (60 mg, 0.15 mmol) (from intermediate 13 above) and racemic 1- A mixture with amino-2-propanol (3 g, 40 mmol) (Aldrich) was heated at 130 ° C. for 16 hours. The mixture was cooled, diluted with ethyl acetate (50 mL) and then washed with water. The aqueous phase was extracted with ethyl acetate (50 mL). The organic phases were separated, combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by chromatography (ethyl acetate: methanol, 85:15) to give rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7- Carboxylic acid (2-hydroxy-propyl) -amide was obtained as a white solid (yield 34 mg, 52%).

_{C 18 H 18 BrN 3 O 3} S + H [(M + H) +] for HRMS m / z calculated: 436.0325. Found: 436.0329.

Example 8
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2,3-dihydroxy-propyl) -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (80 mg, 0.20 mmol) (from intermediate 13 above) and racemic 3- A mixture of amino-1,2-propanediol (3 g, 33 mmol) (Aldrich) was heated at 130 ° C. for 16 hours. The mixture was cooled and diluted with a mixed co-solvent of ethyl acetate and methanol (1: 1, 20 mL). The formed precipitate was filtered, dried and collected to give rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2,3-Dihydroxy-propyl) -amide was obtained as a white solid (yield 80 mg, 90%).

_{C 18 H 18 BrN 3 O 4} S + H [(M + H) +] for HRMS m / z calculated: 452.0274. Found: 452.0279.

Example 9
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1,1-dimethyl-ethyl) -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (80 mg, 0.20 mmol) (from intermediate 13 above) and 2-amino A mixture with 2-methyl-1-propanol (4 g, 45 mmol) (Aldrich) was heated at 130 ° C. for 16 hours. The mixture was cooled, diluted with ethyl acetate (100 mL) and then washed with water. The aqueous phase was extracted with ethyl acetate (100 mL). The organic phases were separated and combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue is purified by chromatography (ethyl acetate: methanol, 10: 1) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-Hydroxy-1,1-dimethyl-ethyl) -amide was obtained as a white solid (yield 30 mg, 33%).

_{C 19 H 20 BrN 3 O 3} S + H [(M + H) +] for HRMS m / z calculated: 450.0482. Found: 450.0487.

Example 10
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1-hydroxymethyl-ethyl) -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (80 mg, 0.20 mmol) (from intermediate 13 above) and 2-amino A mixture with -1,3-propanediol (3 g, 33 mmol) (Aldrich) was heated at 180 ° C. for 5 hours. The mixture was cooled and diluted with a mixed co-solvent of ethyl acetate and methanol (1: 1, 20 mL). The formed precipitate was filtered, dried and collected to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2- Hydroxy-1-hydroxymethyl-ethyl) -amide was obtained as a white solid (yield 60 mg, 66%).

_{C 18 H 18 BrN 3 O 4} S + H [(M + H) +] for HRMS m / z calculated: 452.0274. Found: 452.0279.

Example 11
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-diethylamino-ethyl) -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (20.4 mg, 0.054 mmol) (from intermediate 16 above) and 1-hydroxy -Benzotriazole hydrate (10.0 mg, 0.074 mmol) (Aldrich) and 1,3-diisopropyl-carbodiimide (10.0 μL, 0.064 mmol) (Aldrich) in tetrahydrofuran: N, N-dimethylformamide (1.2 mL, 5: 1) with vigorous stirring. After the reaction material temporarily turned into a solution, the solid precipitated again. The mixture was stirred at room temperature for 30 minutes. N, N-diethylenediamine (15 μL, 0.11 mmol) (Aldrich) was then added and stirred at room temperature overnight. The reaction mixture was concentrated. The residue was taken up in ethyl acetate and washed with water and brine. Any unreacted starting material remained in the aqueous phase. The organic phase was concentrated and purified by reverse phase chromatography (SB-C18 column, 25 mm × 21.2 mm, 5-90% acetonitrile-water (containing 0.75% trifluoroacetic acid) gradient for 10 minutes). Product containing fractions were combined and lyophilized. The lyophilizate (trifluoroacetate) was combined with the equivalent obtained by another test and dissolved in ethyl acetate. The trifluoroacetate salt was neutralized by washing with 1N sodium hydroxide and then with water and brine. The organic phase was dried and concentrated. The residue was recrystallized from ethyl acetate-hexane to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-diethylamino-ethyl). ) -Amide was obtained (yield 32.7 mg, 43.0% (combination yield of the two tests)).

_{C 21 H 25 BrN 4 O 2} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 477.0955. Found: 477.0961.

Example 12a
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (21.1 mg, 0.056 mmol) (from intermediate 16 above) and 1-hydroxy -Benzotriazole hydrate (12.0 mg; 0.089 mmol) (Aldrich) and 1,3-diisopropyl-carbodiimide (12.5 μL, 0.080 mmol) (Aldrich) in tetrahydrofuran: N, N-dimethylformamide (1.2 mL, 5: 1) and mixed with vigorous stirring. The reactant gradually became a solution. After 1 hour, 4-pyrrolidinobutylamine (23.0 mg; 0.16 mmol) (Pfaltz & Bauer) was added and stirring was continued at room temperature. After approximately 40 minutes, the reaction mixture was concentrated. The residue was dissolved in ethyl acetate and washed with water and brine. The organic phase was concentrated and together with the material obtained from the other reactions, 10 of reverse phase HPLC (SB-C18 column, 25 mm × 21.2 mm, 5-90% acetonitrile-water (containing 0.75% trifluoroacetic acid). Purified by a gradient of minutes). Pure fractions obtained from all distributions were combined and lyophilized. The amorphous solid (trifluoroacetate) was dissolved in ethyl acetate and neutralized by 1N sodium hydroxide wash. The organic phase was washed until neutralized with water and brine, dried over sodium sulfate and concentrated. This material was further recrystallized from ethyl acetate-hexane to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidine- 1-yl-butyl) -amide was obtained (yield 11.9 mg).

_{C 23 H 27 BrN 4 O 2} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 503.1111. Found: 503.1114.

Example 12b
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide hydrochloride

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0.187 g; 0.49 mmol) (from intermediate 16 above) and 1-hydroxy Benzotriazole hydrate (0.13 g; 0.084 mmol) (Aldrich) and 1,3-diisopropyl-carbodiimide (0.11 mL, 0.70 mmol) (Aldrich) were added to tetrahydrofuran: N, N-dimethylformamide ( In 48 mL, 5: 1) with vigorous stirring. The solution was stirred at room temperature for 4 hours, after which 4-pyrrolidinobutylamine (0.20 g; 1.41 mmol) (Pfaltz & Bauer) was added and stirring was continued overnight at room temperature. The reaction was concentrated. The residue was partitioned between ethyl acetate and water. The organic phase was washed with water (2x) and brine and then concentrated. The residue was dissolved in an aqueous trifluoroacetic acid solution, the insoluble material was removed by filtration, and then lyophilized. The lyophilized trifluoroacetate salt was diluted with ethyl acetate and neutralized with 1N sodium hydroxide to form the free base, then washed with water and brine. The organic phase was concentrated. The free base residue was dissolved in hot tetrahydrofuran (30 mL) and treated with 1 equivalent of 1N aqueous hydrochloric acid. As a result, hydrochloride precipitated from the solution. The solid was collected, redissolved in water and lyophilized to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4 -Pyrrolidin-1-yl-butyl) -amide hydrochloride was obtained (yield 0.17 g, 65.7%).

_{C 23 H 27 BrN 4 O 2} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 503.1111. Found: 503.1105.

Example 12c
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide methanesulfonate

  This compound can be prepared analogously to the compound of Example 12b using the corresponding amide and methanesulfonic acid.

Example 13a
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholin-4-yl-ethyl) -amide

  N- (2-aminoethyl) -morpholine (2.0 mL) (Aldrich) and methanol (2.0 mL) were combined and stirred with sodium sulfate and basic alumina for 2-3 hours. A portion of this solution (1.0 mL; 3.8 mmol) was added to 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester (36. 3 mg; 0.089 mmol) (from intermediate 13 above) and sodium cyanide (12.0 mg; 0.25 mmol) was added to the mixture. A clear solution was obtained immediately. The solution was heated at 65 ° C. After 3 hours, solids began to precipitate from the solution. The reaction product after 42 hours was analyzed by liquid chromatography. As a result, the desired product and acid by-product (4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine- An approximately 1: 1 mixture with 7-carboxylic acid was confirmed, the reaction was diluted with ethyl acetate, the organic solution was washed with water and brine, dried over sodium sulfate and concentrated. The product was recrystallized from ethyl acetate-hexane to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholine-4- (Il-ethyl) -amide was obtained (yield 11.0 mg, 25.1%).

_{C 21 H 23 BrN 4 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 491.0747. Found: 491.0749.

Example 13b
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholin-4-yl-ethyl) -amide hydrochloride

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (40.7 mg; 0.11 mmol) (from intermediate 16 above) and 1-hydroxy Benzotriazole hydrate (23.8 mg; 0.18 mmol) (Aldrich) and 1,3-diisopropylcarbodiimide (25 μL, 0.16 mmol) (Aldrich) were added to tetrahydrofuran: N, N-dimethylformamide (16 mL, 5 mL). 1) in vigorous stirring. After the solution was stirred at room temperature for 3.75 hours, N- (2-aminoethyl) -morpholine (42 μL; 0.32 mmol) (source) was added and stirring was continued at room temperature. After 40 hours, the reaction was concentrated. The residue was taken up in ethyl acetate and the resulting organic phase was washed with water and brine. The organic phase was concentrated. The residue was dissolved in an aqueous trifluoroacetic acid solution, the insoluble material was removed by filtration, and reverse phase HPLC (SB-C18 column, 25 mm × 21.2 mm, 5-90% acetonitrile-water (containing 0.75% trifluoroacetic acid) ) 10-minute gradient). The pure product was combined and concentrated to near dryness. The residue was diluted with ethyl acetate and neutralized with 1N sodium hydroxide to form the free base, then washed with water and brine. The free base (27.4 mg; 0.056 mmol) was dissolved in hot tetrahydrofuran and treated with 1 equivalent of 1N aqueous hydrochloric acid (53 μL). As a result, hydrochloride precipitated from the solution. The solid was collected and dried to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholin-4-yl- Ethyl) -amide hydrochloride was obtained (yield 16.2 mg, 28.6%).

_{C 21 H 23 BrN 4 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 491.0747. Found: 491.0746. KDR IC ₅₀ 0.0584 μM, FGFR IC ₅₀ 0.1803 μM, VEGF-HUVEC 0.204 μM, FGF-HUVEC 0.627 μM.

Example 14
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (3-dimethylamino-2,2-dimethyl-propyl) -amide

  4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0. 0) in anhydrous N, N-dimethylformamide and acetonitrile (1: 1, 10 mL). 1 g, 0.26 mmol) (from intermediate 16 above) and N, N, 2,2-tetramethyl-1,3-propanediamine (3 eq, 0.1 g, 0.79 mmol) (Aldrich) Diphenylphosphoryl azide (0.29 g, 1.06 mmol) (Aldrich) and triethylamine (3 eq, 0.08 g, 0.79 mmol) (Aldrich) were added. The reaction mixture was stirred at room temperature for 16 hours, then diluted with ethyl acetate (100 mL) and washed with water. The organic phase was separated, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by chromatography (ethyl acetate: methanol: triethylamine, 9: 1: 0.04) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c]. Pyridine-7-carboxylic acid (3-dimethylamino-2,2-dimethyl-propyl) -amide was obtained as a white solid (yield 24 mg, 19%).

_{C 22 H 27 BrN 4 O 2} S + CH 3 OH + H [(M + CH 3 OH + H) +] for HRMS m / z calculated: 523.1373. Found: 523.1347.

Example 15
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (5-diethylamino-1-methyl-pentyl) -amide

  4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0. 0) in anhydrous N, N-dimethylformamide and acetonitrile (1: 1, 10 mL). 1 g, 0.26 mmol) (from intermediate 16 above) and racemic 2-amino-5-diethylaminopentane (0.12 g, 0.79 mmol) (Aldrich) in a solution of diphenylphosphoryl azide (0.29 g, 1.06 mmol). ) (Aldrich) and triethylamine (0.08 g, 0.79 mmol) (Aldrich) were added. The reaction mixture was stirred at room temperature for 16 hours, then diluted with ethyl acetate (100 mL) and washed with water. The organic phase was separated, dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by chromatography (ethyl acetate: methanol: triethylamine, 8: 2: 0.04) to give rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2- c] Pyridine-7-carboxylic acid (4-diethylamino-1-methyl-butyl) -amide was obtained as a white solid (yield 21 mg, 16%).

_{C 24 H 31 BrN 4 O 2} S + H [(M + H) +] for HRMS m / z calculated: 519.1424. Found: 519.1426.

Example 16
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0.1 g, 0.26 mmol) in thionyl chloride (20 mL) (Intermediate 16 above) To) was added a drop of triethylamine (0.1 mL). The reaction mixture was then heated at 80 ° C. for 1 hour. The reaction mixture was cooled and concentrated to dryness. To the residue was added ammonia in methanol (20 mL, 40 mmol, 2N). The reaction mixture was then stirred at room temperature for 24 hours. The mixture is concentrated and the residue is purified by chromatography (ethyl acetate: methanol, 20: 1) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine. -7-carboxylic acid amide was obtained as an off-white solid (yield 35 mg, 36%).

_{C 15 H 12 BrN 3 O 2} S-H 2 [(M-2H) +] for HRMS m / z calculated: 376.9834. Found: 376.99813.

  The compounds of Examples 17-24 can be prepared by following similar procedures as the compounds of Examples 12a and 12b, using the corresponding amines to generate the appropriate compounds unless otherwise illustrated.

Example 17
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0.05 g, 0.13 mmol) (from intermediate 16 above) and 1-hydroxy Benzotriazole hydrate (28.5 mg; 0.21 mmol) (Aldrich), 1,3-diisopropylcarbodiimide (0.03 mL, 0.19 mmol) (Aldrich), tetrahydrofuran and N, N-dimethylformamide (3 6 mL, 5: 1, V / V) with stirring. After 1 hour, 2- (2-pyrrolidin-1-yl-ethoxy) -ethylamine (0.06 g; 0.40 mmol) (from Intermediate 18 above) was added. The mixture was stirred at room temperature for 3 days and then concentrated under reduced pressure. The residue was purified by C18 column chromatography eluting with acetonitrile-water (20-90% gradient) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c Pyridine-7-carboxylic acid [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -amide was obtained as a white powder (yield 36.6 mg, 54%).

_{C 23 H 27 BrN 4 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 519.1060. Found: 519.1060.

Example 18
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (3-methoxy-pyrrolidin-1-yl) -butyl] -amide

Example 19
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-piperidin-1-yl-butyl) -amide

Example 20
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-piperidin-1-yl-ethoxy) -ethyl] -amide

Example 21
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (3-methoxy-piperidin-1-yl) -butyl] -amide

Example 22
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-morpholin-4-yl-butyl) -amide

  4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0.05 g, 0.13 mmol) in thionyl chloride (20 mL) (Aldrich) (above To a solution of intermediate 16), a drop of triethylamine (0.1 mL) was added. The reaction mixture was then heated at 80 ° C. for 1 hour. The reaction mixture was cooled and concentrated to dryness under reduced pressure. To the residue was added 4- (4-aminobutyl) morpholine (0.06 g, 0.40 mmol) (from Intermediate 17 above). The mixture was stirred at room temperature for 18 hours and concentrated under reduced pressure.

  The residue was purified by C18 column chromatography eluting with acetonitrile-water (40-80% gradient) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c Pyridine-7-carboxylic acid (4-morpholin-4-yl-butyl) -amide was obtained (yield 34 mg, 51%).

_{C 23 H 27 BrN 4 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 519.1060. Found: 519.1060.

Example 23
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-morpholin-4-yl-ethoxy) -ethyl] -amide

Example 24
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (4-methoxy-piperidin-1-yl) -butyl] -amide

  4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0.05 g, 0.13 mmol) (from intermediate 16 above) and 1-hydroxy Benzotriazole hydrate (28.5 mg, 0.21 mmol) (Aldrich), 1,3-diisopropylcarbodiimide (0.03 mL, 0.19 mmol) (Aldrich), tetrahydrofuran and N, N-dimethylformamide (3 .6 mL, 5: 1, V / V) in a mixed solution with stirring. After 1 hour, 4- (4-methoxy-piperidin-1-yl) -butylamine (0.07 g; 0.40 mmol) (from Intermediate 19 above) was added. The mixture was stirred at room temperature for 3 days and then concentrated under reduced pressure. The residue was purified by C18 column chromatography eluting with acetonitrile-water (20-90% gradient) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c ] Pyridine-7-carboxylic acid [2- (2-piperidin-1-yl-ethoxy) -ethyl] -amide was obtained as a white powder (yield 22.0 mg, 31%).

_{C 25 H 31 BrN 4 O 3} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 547.1373. Found: 547.1372.

Example 25
4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [3- (2,3-dihydroxy-propoxy) -propyl] -amide

4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (0.05 g, 0.13 mmol) (from intermediate 16 above) and 1-hydroxy Benzotriazole hydrate (28.5 mg; 0.21 mmol) (Aldrich), 1,3-diisopropylcarbodiimide (0.03 mL, 0.19 mmol) (Aldrich), tetrahydrofuran and N, N-dimethylformamide (3 .6 mL, 5: 1, V / V) in a mixed solution with stirring. After 1 hour, 3- (2,2-dimethyl- [1,3] dioxolan-4-yl-methoxy) -propylamine (75.7 mg, 0.40 mmol) (from Intermediate 20 above) was added. The mixture was stirred at room temperature for 18 hours and then concentrated under reduced pressure. The residue was diluted with ethyl acetate, washed with water and brine, dried (MgSO ₄ ), filtered and concentrated under reduced pressure. The residue was purified by flash chromatography eluting with hexane-ethyl acetate (80-100% gradient) to give 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c Pyridine-7-carboxylic acid [3- (2,2-dimethyl- [1,3] dioxolan-4-ylmethoxy) -propyl] -amide was obtained (yield 60 mg, 84%).

  4-Amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [3- (2,2-dimethyl- [1,3]] in ethanol (2 mL) To a solution of dioxolan-4-ylmethoxy) -propyl] -amide (60.0 mg, 0.11 mmol) was added 1N aqueous hydrochloric acid (2 mL). The mixture was stirred at room temperature for 18 hours. The mixture was then cooled in an ice-water bath and 1N sodium hydroxide solution (2 mL) was added. Ethanol was then removed under reduced pressure. The residue was washed with water and purified by flash chromatography eluting with 10% ethanol in ethyl acetate to give rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3, 2-c] pyridine-7-carboxylic acid [3- (2,3-dihydroxy-propoxy) -propyl] -amide was obtained as a white powder (yield 32 mg, 57%).

_{C 21 H 24 BrN 3 O 5} S + H [(M + H) +] for HRMS (ES +) m / z calcd: 510.0693. Found: 510.0693.

  The antiproliferative activity of the compounds of the present invention is demonstrated by Examples 26 and 27 below. These activities are useful for the compounds of the present invention to treat cancers, especially solid tumors, especially breast, lung, prostate and colon cancerous solid tumors, especially breast and colon cancerous solid tumors. It shows that.

Example 26
Kinase assay

  To confirm inhibition of KDR and FGFR, a kinase assay was performed using an HTRF (homogeneous time-resolved fluorescence) assay. This assay is described in A. J. Kolb et al., Drug Discovery Today, 1998, 3 (7), pp.333.

Prior to the kinase reaction, EEE-tagged KDR was added in the presence of activation buffer (50 mM HEPES, pH 7.4, 1 mM DTT, 10% glycerol, 150 mM NaCl, 0.1 mM EDTA, 26 mM MgCl ₂ , and 4 mM ATP). Activated with. The enzyme was incubated for 1 hour at 4 ° C.

Kinase activity assays were performed using 96 well polypropylene plates (Falcon) with a total volume of 90 μL in each well. Each well contains 1 μM KDR substrate (Biotin-EEEEEYFELVAKKKK), 1 nM activated KDR, and any one of eight assay concentrations from 100 μM to 128 μM (1: 5 serial dilution). It was. Kinase activity assays consisted of 100 mM HEPES, pH 7.4, 1 mM DTT, 0.1 mM Na ₂ VO ₄ , 25 mM MgCl ₂ , 50 mM NaCl (from KDR stock solution), 1% DMSO (from compound), 0. 3mM of ATP (K _m concentration) and was carried out in the presence of 0.02% BSA. The reaction was incubated at 37 ° C. for 30 minutes. To stop the KDR reaction, 72 μL of the reaction mixture was diluted with 18 μL of exposure buffer (20 mM EDTA, 50 mM HEPES, pH 7.4, 0.02% BSA, 10 nM Eu labeled anti-pY antibody (final concentration 2 nM ), And 100 nM streptavidin (final concentration 20 nM)). After mixing, 35 μL of each solution was placed in two wells of a 384 well black plate (Costar) and read at 615/665 nm using a Wallac Vitor 5 reader.

The FGFR activity assay was performed in the same manner as the KDR activity assay procedure described above, with the following differences. GST-labeled FGFR enzyme was activated for 1 hour at room temperature in an activation buffer containing 100 mM HEPES, pH 7.4, 50 mM NaCl, 20 mM MgCl ₂ , and 4 mM ATP. The kinase activity assay was performed in 100 mM HEPES, 1 mM DTT, 0.4 mM MgCl ₂ , 0.4 mM MnCl ₂ , 50 mM NaCl, 1% DMSO, 10 μM ATP (K _{m for} FGFR) in a total volume of 90 μL. = 8.5 μM), 0.1 mM Na ₂ VO ₄ , and 0.02% BSA in the presence of 1 μM substrate (Biotin-EEEEEYFELV), 1.5 nM activated FGFR, and test compound. Carried out. Other aspects of the assay were performed as in the KDR assay.

Compound IC ₅₀ values are calculated by fitting data to the formula Y = [(ab) / {1+ (X / c) ^d ] + b using Excel® with double data groups did. In the above formula, a and b are the enzyme activity in the absence of the test inhibitor compound and the enzyme activity in the presence of an infinite amount of the inhibitor test compound, respectively, c is IC ₅₀ , d is the compound The hill constant of the response. The IC ₅₀ value is the test compound concentration that reduces enzyme activity by 50% under the test conditions.

The compounds of the present invention have a KDR IC ₅₀ value of less than 5 μM, preferably less than 1.5 μM. Alternatively, the compounds of the invention have an FGFR IC ₅₀ value of less than 5 μM, preferably less than 2.5 μM. Among them, the compound of the present invention most preferably has a KDR IC ₅₀ value of less than 1.5 μM and an FGFR IC ₅₀ value of less than 2.5 μM.

Example 27
HUVEC proliferation assay stimulated by VEGF and FGF

The anti-proliferative activity of the test compounds of the present invention in a cell-based assay was evaluated by the BrdU assay using a BrdU kit (Roche Biochemicals 1-647-229). Human umbilical vein endothelial cells (HUVEC, Clonetics CC-2519) were cultured overnight in EGM-2 (Clonetics CC-3162) medium and EGM-2 (Clonetics CC-3162) in 96 well flat bottom plates (Costar 3595) Seeded at 10,000 cells per well in a 200 μL volume of medium. After growth for 24 hours at 37 ° C. under 5% CO ₂ , the incubation medium was slowly removed by aspiration and the contents of each well were removed with 50 μg gentamicin per mL and 50 ng amphotericin-B (Clonetics CC-B / mL). 4083), and washed with 300 μL of warmed EBM-2 (Clonetics CC-3156). Subsequently, the remaining medium was aspirated again, instead, 160 μL of serum starvation medium per well (1% heat inactivated FBS in EBM-2 (Clonetics CC-4102), 50 μg gentamicin per mL and 1 mL 50 ng amphotericin-B (Clonetics CC-4083), 10 units per mL Wyeth-Ayerst heparin (NDC0641-0391-25), and 2 mM L-glutamine (supplemented with GIBCO 25030-081)). After the cells were serum starved for 24 hours, 20 μL of a solution containing 10% of the test compound in serum starvation medium containing 2.5% DMSO was added to the appropriate wells. Control wells received 20 μL of serum starvation medium containing 2.5% DMSO. Plates were returned to the incubator for 2 hours. Cells are preincubated with test compound for 2 hours, followed by 20 μL of 10 × assay concentration of growth factor diluted in serum starvation medium, 50 ng FGF per mL, or 200 ng VEGF per mL (R & D systems 293-VE) Was added. The final concentration of FGF in the assay was 5 ng / mL and the final concentration of VEGF in the assay was 20 ng / mL. Control wells containing no growth factor received 20 μL of serum starvation medium containing the same amount of BSA as wells containing growth factor. Plates were returned to the incubator for an additional 22 hours.

BrdU ELISA
After 24 hours of exposure to the test compound, 20 μL of BrdU labeling reagent diluted in serum starvation medium (1: 100) was added per well and the cells were labeled with BrdU (Roche Biochemicals 1-647-229). The plate was then returned to the incubator for 4 hours. The medium was removed on a paper towel and the labeling medium was removed. 200 μL of fixing / denaturing solution was added to each well and incubated at room temperature for 45 minutes to fix the cells and denature the DNA. The fixative / denaturation solution was removed onto a paper towel, 100 μL of anti-BrdU-POD was added to each well, and the wells were incubated at room temperature for 2 hours. The antibody solution was removed, and the wells were washed 3 or 4 times with 300 μL of PBS each. 100 μL of TMB substrate solution was added to each well and the wells were incubated at room temperature for 5-8 minutes. The reaction was then stopped by adding 100 μL of 1M phosphoric acid per well. The plate was read at 450 nm with a reference wavelength of 650 nm. The percent inhibition for each test compound was obtained by subtracting the absorbance of blank (no cell) wells from the absorbance of all wells, then dividing the average absorbance of duplicate tests by the mean value of the control. The obtained value was subtracted from 1 and multiplied by 100 (inhibition rate% = (1−average absorbance of double run test / average value of control) × 100). IC ₅₀ value is the concentration of test compound that inhibits BrdU labeling by 50% and is a measure of cell growth inhibition. IC ₅₀ values are determined from linear regression of a logarithmic plot of concentration versus percent inhibition.

The compounds of the invention have a VEGF stimulated HUVEC assay IC ₅₀ value of less than 3 μM, preferably less than 1.5 μM. Alternatively, the compounds of the present invention have an FGF stimulated HUVEC assay IC ₅₀ value of less than 5 μM, preferably less than 3.0 μM, more preferably less than 2 μM. Among them, the compound of the present invention most preferably has a VEGF stimulated HUVEC assay IC ₅₀ value of less than 1.5 μM and an FGF stimulated HUVEC assay IC ₅₀ value of less than 2 μM.

Example 28
Tablet preparation

Manufacturing procedure :
1. Mix items 1, 2 and 3 with a suitable mixer for 15 minutes.
2. Granulate the mixed powder obtained in step 1 with 20% povidone K30 solution (item 4).
3. The granules obtained in step 2 are dried at 50 ° C.
4). Treat the granules obtained in step 3 with a suitable crusher.
5. Add item 5 to the crushed granules obtained in step 4 and mix for 3 minutes.
6). Compress the granules obtained in step 5 with a suitable press.

Example 29
Capsule preparation

Manufacturing procedure :
1. Mix items 1, 2 and 3 with a suitable mixer for 15 minutes.
2. Add items 4 and 5 and mix for 3 minutes.
3. Fill into suitable capsules.

Example 30
Preparation of injection / emulsion

Manufacturing procedure :
1. Item 1 is dissolved in Item 2.
2. Add items 3, 4 and 5 to item 6, mix and disperse, then homogenize.
3. Add the solution obtained in step 1 to the mixture obtained in step 2 and homogenize until the dispersion is translucent.
4. Sterile filter through 0.2 μm filter and fill into vials.

Example 31
Preparation of injection / emulsion

Manufacturing procedure :
1. Item 1 is dissolved in Item 2.
2. Add items 3, 4 and 5 to item 6, mix until dispersed, then homogenize.
3. Add the solution obtained in step 1 to the mixture obtained in step 2 and homogenize until the dispersion is translucent.
4. Sterile filter through 0.2 μm filter and fill into vials.

  Although the present invention has been described with reference to specific and preferred embodiments, it will be apparent to those skilled in the art that changes and modifications may be made through routine experimentation and practice of the invention. In other words, the present invention is not limited to the above description, but is intended to be defined in the appended claims and their equivalents.

Claims (48)

formula

A compound of the formula
R ¹ is substituted with lower alkyl and OR ³ , NR ³ R ⁴ , S (O) _n R ³ , cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, heteroaryl, or substituted heteroaryl Selected from lower alkyl
R ² is
H,
Lower alkyl, and ^{OR 5, OC (O) R} 5, NR 5 R 6, S (O) n R 5, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, Selected from aryl, or lower alkyl substituted with substituted heteroaryl;
R ³ and R ⁴ are independently
H,
Lower alkyl,
Aryl, aryl fused to a heterocycle or substituted heterocycle, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, or lower alkyl substituted with a substituted heterocycle,
Aryl,
Aryl fused to a heterocycle or substituted heterocycle,
Substituted aryl,
Heteroaryl,
Heteroaryl fused to a heterocycle or substituted heterocycle,
Substituted heteroaryl,
Heterocycle,
A heterocycle fused to an aryl,
Selected from cycloalkyl, and substituted cycloalkyl;
Alternatively, the group NR ³ R ⁴ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ³ and R ⁴ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally having one or more lower alkyl, ═O, OR ⁷ , Optionally substituted by a group consisting of COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ ;
R ⁵ and R ⁶ are independently
H,
Selected from lower alkyl and lower alkyl substituted with OR ⁷ , NR ⁷ R ⁸ , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl,
Alternatively, the group NR ⁵ R ⁶ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ⁵ and R ⁶ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally having one or more lower alkyl, ═O, OR ⁷ , Optionally substituted by a group consisting of NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ ;
R ⁷ and R ⁸ are independently
Selected from H and lower alkyl, or
The group NR ⁷ R ⁸ may independently form a ring having 3 to 7 total atoms, and the ring atom contains a carbon ring atom in addition to the nitrogen to which R ⁷ and R ⁸ are bonded. Wherein the carbon ring atom is optionally substituted by one or more additional heteroatoms, and the ring atom is optionally from one or more lower alkyl, ═O, or OR ^9. Optionally substituted by the group
R ⁹ is H or lower alkyl;
n is 0, 1, or 2;
here,
Substituted aryl and substituted heteroaryl, lower ^{alkyl, OR 7, NR 7 R 8} , COR 7, CO 2 R 7, CONR 7 R 8, SO 2 NR 7 R 8, SO n R 7, CN, NO 2 And aryl and heteroaryl substituted with one or more groups independently selected from halogen;
Substituted cycloalkyl and substituted heterocycle are lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO ₂ NR ⁷ R ⁸ , SO _n R ⁷ , And cycloalkyl and heterocycle substituted with one or more groups independently selected from CN), or a pharmaceutically acceptable salt or ester thereof. The compound of claim 1, wherein R ¹ is lower alkyl substituted with OR ³ . The compound of claim 2, wherein R ³ is selected from aryl, aryl substituted with halogen, and aryl fused to a heterocycle.   4. The compound of claim 3, wherein the halogen is Br, Cl, or F. R ³ groups are heteroaryl, and is selected from heteroaryl optionally substituted with OR ^7, compound of claim 2. 2. The compound of claim 1, wherein R < ¹ > is lower alkyl substituted with NR < ³ > R < ⁴ >. The group NR ³ R ⁴ forms a ring with a total of 3 to 7 ring atoms comprising carbon ring atoms in addition to the nitrogen to which R ³ and R ⁴ are bonded, the carbon ring atoms optionally being 1 Or optionally substituted by two or more further heteroatoms, wherein said ring atom is optionally one or more lower alkyl, ═O, OR ⁷ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ^7, and SO ₂ NR ⁷ R ^8, preferably may be substituted by a group consisting of oR ^7, compound of claim 6. R ¹ is lower alkyl substituted with S (O) _n R ³ , and R ³ is condensed to lower alkyl, heterocycle, heterocycle fused to aryl, aryl, substituted aryl, and heterocycle 2. A compound according to claim 1 which is aryl. The compound of claim 1, wherein R ¹ is lower alkyl substituted with cycloalkyl. The compound according to claim 1, wherein R ¹ is lower alkyl substituted with substituted cycloalkyl. The compound according to claim 1, wherein R ¹ is lower alkyl substituted with a heterocycle. The compound according to claim 1, wherein R ¹ is lower alkyl substituted with a substituted heterocycle. The compound of claim 1, wherein R ¹ is lower alkyl substituted with heteroaryl. The compound according to claim 1, wherein R ¹ is lower alkyl substituted with substituted heteroaryl. The compound according to claim 1, wherein R ¹ is lower alkyl. The compound of claim 1, wherein R ² is lower alkyl substituted with OR ⁵ , and R ⁵ is lower alkyl substituted with NR ⁷ R ⁸ . The compound according to claim 1, wherein R ² is lower alkyl substituted with NR ⁵ R ⁶ . The compound of claim 1 wherein R ² is lower alkyl substituted with OR ⁵ and R ⁵ is hydrogen. The compound according to claim 1, wherein R ² is lower alkyl substituted with OR ⁵ , and R ⁵ is lower alkyl substituted with OR ⁷ . The group NR ⁵ R ⁶ forms, in addition to the nitrogen to which R ⁵ and R ⁶ are attached, a ring with a total of 3 to 7 ring atoms including carbon ring atoms, said carbon ring atoms optionally being 1 or 2 The ring atom may be optionally substituted by the above further heteroatoms, and the ring atom is optionally one or more lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ^7. R ^8, SO _n R ^7, and SO ₂ NR ⁷ may be substituted by a group consisting of R ^8, claim 17 a compound according. The compound of claim 1, wherein R ² is lower alkyl substituted with SO _(n) R ⁵ . The compound according to claim 1, wherein R ² is lower alkyl. The compound of claim 1, wherein R ² is lower alkyl substituted with one OH group or one NR ⁵ R ⁶ group. The compound of claim 1, wherein R ² is H. Formula I

A compound of the formula
R ¹ is lower alkyl substituted with OR ³ ;
R ² is H or lower alkyl substituted with one OR ⁵ group or one NR ⁵ R ⁶ group;
R ³ is aryl substituted with halogen or OR ⁷ , or aryl fused to a heterocycle;
R ⁵ and R ⁶ are independently H or lower alkyl, or the group NR ⁵ R ⁶ may independently form a ring having a total number of 3 to 6, wherein the ring atom is In addition to the nitrogen to which R ⁵ and R ⁶ are bonded, it comprises a carbocyclic atom, said carbocyclic atom optionally being substituted by one further heteroatom selected from N or O The ring atom may optionally be substituted by OR ⁷ ;
R ⁷ is H or lower alkyl), or a pharmaceutically acceptable salt or ester thereof. 4-amino-3- (4-bromo-2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide;
4-amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide;
4-amino-3- (benzo [1,3] dioxol-5-yloxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide, and 4-amino-3-phenoxymethyl-thieno The compound of claim 1 selected from the group consisting of [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-ethyl) -amide. rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1-methyl-ethyl) -amide;
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-propyl) -amide, and rac-4-amino-3- The compound of claim 1 selected from the group consisting of (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2,3-dihydroxy-propyl) -amide. 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1,1-dimethyl-ethyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-hydroxy-1-hydroxymethyl-ethyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-diethylamino-ethyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amide hydrochloride, and 4-amino- Selected from the group consisting of 3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-pyrrolidin-1-yl-butyl) -amidomethanesulfonate The compound of claim 1. 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholin-4-yl-ethyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (2-morpholin-4-yl-ethyl) -amide hydrochloride;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (3-dimethylamino-2,2-dimethyl-propyl) -amide;
rac-4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (5-diethylamino-1-methyl-pentyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid amide, and 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3 , 2-c] Pyridine-7-carboxylic acid [2- (2-pyrrolidin-1-yl-ethoxy) -ethyl] -amide. 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (3-methoxy-pyrrolidin-1-yl) -butyl] -amide,
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-piperidin-1-yl-butyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-piperidin-1-yl-ethoxy) -ethyl] -amide,
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (3-methoxy-piperidin-1-yl) -butyl] -amide,
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid (4-morpholin-4-yl-butyl) -amide;
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [2- (2-morpholin-4-yl-ethoxy) -ethyl] -amide, and 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [4- (4-methoxy-piperidin-1-yl) -butyl] -amide,
From the group consisting of 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid [3- (2,3-dihydroxy-propoxy) -propyl] -amide 2. A compound according to claim 1 which is selected.   A pharmaceutical composition comprising a compound of formula I according to claim 1 together with a pharmaceutically acceptable adjuvant. Therapeutically effective amount of formula I

A compound of the formula
R ¹ is substituted with lower alkyl and OR ³ , NR ³ R ⁴ , S (O) _n R ³ , cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, heteroaryl, or substituted heteroaryl Selected from lower alkyl
R ² is H, lower alkyl, and OR ⁵ , OC (O) R ⁵ , NR ⁵ R ⁶ , S (O) _n R ⁵ , aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclic ring, Selected from substituted heterocycle, heteroaryl, or lower alkyl substituted with substituted heteroaryl;
R ³ and R ⁴ are independently
H,
Lower alkyl,
Aryl, aryl fused to a heterocycle or substituted heterocycle, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, or lower alkyl substituted with a substituted heterocycle,
Aryl,
Aryl fused to a heterocycle or substituted heterocycle,
Substituted aryl,
Heteroaryl,
Heteroaryl fused to a heterocycle or substituted heterocycle,
Substituted heteroaryl,
Selected from cycloalkyl, and substituted cycloalkyl;
Alternatively, the group NR ³ R ⁴ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ³ and R ⁴ are bonded. Wherein the carbon ring atom is optionally substituted by one or more additional heteroatoms, and the carbon ring atom is optionally one or more lower alkyl, ═O, OR ^7. , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ may be substituted;
R ⁵ and R ⁶ are independently
H,
Selected from lower alkyl and lower alkyl substituted with OR ⁷ , NR ⁷ R ⁸ , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl,
Alternatively, the group NR ⁵ R ⁶ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ⁵ and R ⁶ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally having one or more lower alkyl, ═O, OR ⁷ , Optionally substituted by a group consisting of NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ ;
R ⁷ and R ⁸ are independently
Selected from H and lower alkyl;
Alternatively, the group NR ⁷ R ⁸ may independently form a ring having a total number of 3-7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ⁷ and R ⁸ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally being one or more lower alkyl, ═O, or OR ^9. Optionally substituted by a group consisting of
R ⁹ is H or lower alkyl;
n is 0, 1, or 2;
Substituted aryl and substituted heteroaryl, lower ^{alkyl, OR 7, NR 7 R 8} , COR 7, CO 2 R 7, CONR 7 R 8, SO 2 NR 7 R 8, SO n R 7, CN, NO 2 And aryl and heteroaryl substituted with one or more groups independently selected from halogen;
Substituted cycloalkyl and substituted heterocycle are lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO ₂ NR ⁷ R ⁸ , SO _n R ⁷ , And cycloalkyl and heterocycles substituted with one or more groups independently selected from CN), or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier or additive. A pharmaceutical composition comprising a dosage form.   A method of treating cancer comprising administering a therapeutically effective amount of a compound of formula I according to claim 1 to a patient in need of such treatment. Therapeutically effective amount of formula I

(Where
R ¹ is substituted with lower alkyl and OR ³ , NR ³ R ⁴ , S (O) _n R ³ , cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, heteroaryl, or substituted heteroaryl Selected from lower alkyl
R ² is
H,
Lower alkyl, and ^{OR 5, OC (O) R} 5, NR 5 R 6, S (O) n R 5, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, Selected from aryl, or lower alkyl substituted with substituted heteroaryl;
R ³ and R ⁴ are independently
H,
Lower alkyl and substituted with aryl, aryl fused to a heterocyclic or substituted heterocycle, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, or substituted heterocycle Lower alkyl,
Aryl,
Aryl fused to a heterocycle or substituted heterocycle,
Substituted aryl,
Heteroaryl,
Heteroaryl fused to a heterocycle or substituted heterocycle,
Substituted heteroaryl,
Selected from cycloalkyl, and substituted cycloalkyl;
Alternatively, the group NR ³ R ⁴ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ³ and R ⁴ are bonded. Wherein the carbon ring atom is optionally substituted by one or more additional heteroatoms, and the carbon ring atom is optionally one or more lower alkyl, ═O, OR ^7. , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ may be substituted;
R ⁵ and R ⁶ are independently
H,
Selected from lower alkyl and lower alkyl substituted with OR ⁷ , NR ⁷ R ⁸ , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl,
Alternatively, the group NR ⁵ R ⁶ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ⁵ and R ⁶ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally having one or more lower alkyl, ═O, OR ⁷ , Optionally substituted by a group consisting of NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ ;
R ⁷ and R ⁸ are independently
Selected from H and lower alkyl;
Alternatively, the group NR ⁷ R ⁸ independently, the ring atom forms a ring with 3-7 total atoms comprising carbon ring atoms in addition to the nitrogen to which R ⁷ and R ⁸ are bonded. The carbon ring atom may optionally be substituted by one or more additional heteroatoms, and the ring atom may optionally be from one or more lower alkyl, ═O, or OR ^9. Optionally substituted by the group
R ⁹ is H or lower alkyl;
n is 0, 1, or 2;
Substituted aryl and substituted heteroaryl, lower ^{alkyl, OR 7, NR 7 R 8} , COR 7, CO 2 R 7, CONR 7 R 8, SO 2 NR 7 R 8, SO n R 7, CN, NO 2 And aryl and heteroaryl substituted with one or more groups independently selected from halogen;
Substituted cycloalkyl and substituted heterocycle are lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO ₂ NR ⁷ R ⁸ , SO _n R ⁷ , And cycloalkyl and heterocycle substituted with one or more groups independently selected from CN), or a pharmaceutically acceptable salt or ester thereof, to a patient in need of such treatment A method for treating cancer comprising the steps. Therapeutically effective amount of formula I

A compound of the above formula
R ¹ is substituted with lower alkyl and OR ³ , NR ³ R ⁴ , S (O) _n R ³ , cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, heteroaryl, or substituted heteroaryl Selected from lower alkyl
R ² is
H,
Lower alkyl, and ^{OR 5, OC (O) R} 5, NR 5 R 6, S (O) n R 5, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, Selected from aryl, or lower alkyl substituted with substituted heteroaryl;
R ³ and R ⁴ are independently
H,
Lower alkyl and substituted with aryl, aryl fused to a heterocyclic or substituted heterocycle, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycle, or substituted heterocycle Lower alkyl,
Aryl,
Aryl fused to a heterocycle or substituted heterocycle,
Substituted aryl,
Heteroaryl,
Heteroaryl fused to a heterocycle or substituted heterocycle,
Substituted heteroaryl,
Selected from cycloalkyl, and substituted cycloalkyl;
Alternatively, the group NR ³ R ⁴ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ³ and R ⁴ are bonded. Wherein the carbon ring atom is optionally substituted by one or more additional heteroatoms, and the carbon ring atom is optionally one or more lower alkyl, ═O, OR ^7. , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ may be substituted;
R ⁵ and R ⁶ are independently
H,
Selected from lower alkyl and lower alkyl substituted with OR ⁷ , NR ⁷ R ⁸ , aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, cycloalkyl, substituted cycloalkyl,
Alternatively, the group NR ⁵ R ⁶ may independently form a ring having a total number of 3 to 7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ⁵ and R ⁶ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally having one or more lower alkyl, ═O, OR ⁷ , Optionally substituted by a group consisting of NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO _n R ⁷ , and SO ₂ NR ⁷ R ⁸ ;
R ⁷ and R ⁸ are independently
Selected from H and lower alkyl;
Alternatively, the group NR ⁷ R ⁸ may independently form a ring having a total number of 3-7, wherein the ring atom includes a carbon ring atom in addition to the nitrogen to which R ⁷ and R ⁸ are bonded. Comprising, said carbon ring atom optionally substituted by one or more further heteroatoms, said ring atom optionally being one or more lower alkyl, ═O, or OR ^9. Optionally substituted by a group consisting of
R ⁹ is H or lower alkyl;
n is 0, 1, or 2;
Substituted aryl and substituted heteroaryl, lower ^{alkyl, OR 7, NR 7 R 8} , COR 7, CO 2 R 7, CONR 7 R 8, SO 2 NR 7 R 8, SO n R 7, CN, NO 2 And aryl and heteroaryl substituted with one or more groups independently selected from halogen;
Substituted cycloalkyl and substituted heterocycle are lower alkyl, ═O, OR ⁷ , NR ⁷ R ⁸ , COR ⁷ , CO ₂ R ⁷ , CONR ⁷ R ⁸ , SO ₂ NR ⁷ R ⁸ , SO _n R ⁷ , And cycloalkyl and heterocycle substituted with one or more groups independently selected from CN), or a pharmaceutically acceptable salt or ester thereof, to a patient in need of such treatment A method for controlling cancer, comprising a step.   34. The method of claim 33, wherein the cancer is breast cancer, lung cancer, colon cancer, or prostate cancer.   36. The method of claim 35, wherein the cancer is breast cancer, lung cancer, colon cancer, or prostate cancer. 3-methyl-5H-thieno [3,2-c] pyridin-4-one,
7-iodo-3-methyl-5H-thieno [3,2-c] pyridin-4-one,
3-methyl-4-oxo-4,5-dihydro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester,
4-chloro-3-methyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester,
3-bromomethyl-4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester,
3- (4-Bromo-2,6-difluoro-phenoxymethyl) -4-chloro-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester, and 4-amino-3- (4-bromo- A compound selected from the group consisting of 2,6-difluoro-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester. 4-Amino-3- (2-chloro-4-methoxy-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester 3- (4-bromo-phenoxymethyl) -4-chloro- Thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester,
4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester,
4-chloro-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester,
4-amino-3-phenoxymethyl-thieno [3,2-c] pyridine-7-carboxylic acid ethyl ester, and 4-amino-3- (4-bromo-phenoxymethyl) -thieno [3,2-c] A compound selected from the group consisting of pyridine-7-carboxylic acid. R ¹ is 4-bromo-phenoxymethyl;
R ² represents the meaning defined in claim 25;
26. A compound of formula I according to claim 25. A process for preparing a compound of formula I according to claim 1, comprising:
a) Formula A:

A compound of formula B:

Converting to a compound of
b) further comprising reacting the compound of formula B in the presence of a compound of formula R ² NH ₂ to form a compound of formula I;
In the above formula,
R ¹ and R ² represent the meaning as defined in claim 1;
The method wherein R ¹⁰ is lower alkyl.   Use of a compound according to claim 1 for the treatment and / or control of cancer.   Use of a compound according to claim 1 for the treatment and / or control of solid tumors.   Use of a compound according to claim 1 for the treatment and / or control of breast cancer, lung cancer, colon cancer or prostate cancer.   Use of a compound according to claim 1 for the manufacture of a medicament for the treatment of cancer.   Use of a compound according to claim 1 for the manufacture of a solid tumor therapeutic agent.   Use of a compound according to claim 1 for the manufacture of a medicament for the treatment of breast cancer, lung cancer, colon cancer or prostate cancer.   The invention described in the attached specification.