Patent Application
20070275962
The present invention relates to heterobicyclic compounds and pharmaceutically acceptable salts thereof, the use of these compounds for the prophylaxis and/or treatment of various diseases such as infectious diseases, including mycobacteria-induced infections and opportunistic diseases, prion diseases, immunological diseases, autoimmune diseases, bipolar and clinical disorders, cardiovascular diseases, cell proliferative diseases, diabetes, inflammation, transplant rejections, erectile dysfunction, neurodegenerative diseases and stroke, as well as compositions containing at least one heterobicyclic compound and/or pharmaceutically acceptable salts thereof. Furthermore, reaction procedures for the synthesis of said heterobicyclic compounds are disclosed.
Object of the present invention is to provide pharmaceutically active compounds for prophylaxis and treatment of various diseases such as infections, inflammations, immunological diseases, cardiovascular diseases, cell proliferative diseases, transplant rejections, or neurodegenerative diseases, methods for the synthesis of said compounds and pharmaceutical compositions containing at least one pharmaceutically active compound.
This object is solved by the heterobicyclic compound according to claim 1 and/or pharmaceutically acceptable salts of said compounds, the use of at least one of those compounds and/or the pharmaceutically acceptable salts thereof as pharmaceutically active agents according to independent claim 8, the use of the compounds as an inhibitor for a protein kinase according to independent claim 9, the use of the compounds for prophylaxis and/or treatment of various diseases according to independent claim 19, the pharmaceutical composition according to independent claim 38, and a method of amidation of a carboxylic acid ester according to independent claim 39. Further advantageous features, aspects and details of the invention are evident from the dependent claims, the description, the examples and the drawings.
The present invention relates to compounds of the general formula (I)
wherein
X1 is selected from S, O, NH, NR4′;
Y1—Y2—Y3—Y4 represent the following residues:
R1, R4, R4′, R4″, R4′″, R4″″, R8, R9 and R15 are independently of each other selected from —H, substituted or unsubstituted C1-C8-alkyl, substituted or unsubstituted C3-C10-cycloalkyl, substituted or unsubstituted C1-C6-heterocyclyl, substituted or unsubstituted C2-C6-alkenyl, substituted or unsubstituted C2-C6-alkinyl, -Ph, —CH2Ph, substituted phenyl, substituted benzyl, substituted or unsubstituted C1-C8-acyl;
R2 represents the residue —CO—NH—R4, —CS—NH—R4, —SO2—NH—R4, —C(═NH)—NH—R4, —COOR4, —SO3—R4;
R3 is selected from —H, —CO—R5, —CS—R5, —C(═NR4″)—R5, —SO2R5, —CO—R5′, —CS—R5′, —C(═NR4″)—R5′, —SO2R5′, —CO—NR4″—R5, —CS—NR4″—R5, —CO—CO—NR4″—R5, —CO—CO—O—R5, —CO—OR5, —CS—OR5, —C(═NH)NR4″—R5;
R5 is selected from substituted or unsubstituted C3-C10-cycloalkyl, substituted or unsubstituted C1-C8-alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6-heterocyclyl, substituted or unsubstituted C2-C6-alkenyl, substituted or unsubstituted alkylaryl, substituted or unsubstituted C2-C6-alkinyl, adamantyl, —H, —R14, —R18, —(CH2)n—C1-C6-heterocyclyl, —(CH2)n—C3-C10-cycloalkyl,
R6, R6′, R7 and R7′ are independently of each other selected from —H, substituted or unsubstituted C3-C10-cycloalkyl, substituted or unsubstituted C1-C8-alkyl, substituted or unsubstituted aryl, -Ph, —CH2Ph, substituted phenyl, substituted benzyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6-heterocyclyl, substituted or unsubstituted C2-C6-alkenyl, substituted or unsubstituted C2-C6-alkinyl, adamantyl;
—R5′, R10, R11, R12, R13, R16 and R17 are independently of each other selected from —H, —OH, —NH2, —R14, —R14′, —R14″, —R14′″, —R14″″, —R18, —R18′, —R18″, —R18′″, —R18″″, -Ph, —CH2Ph, substituted phenyl, substituted benzyl, substituted or unsubstituted C1-C8-alkyl, substituted or unsubstituted C3-C10-cycloalkyl, substituted or unsubstituted C1-C6-heterocyclyl, substituted or unsubstituted C2-C6-alkenyl, substituted or unsubstituted C2-C6-alkinyl, adamantyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6-alkoxyl, substituted or unsubstituted —O—C3-C10-cycloalkyl, substituted or unsubstituted —O—C1-C6-heterocyclyl, substituted or unsubstituted —O-aryl, substituted or unsubstituted —O-heteroaryl, —OOC—O—R14, —O—CO—R14′, —COO—R14′, —O—CO—NR7R14, —NR14″″—CO—NR7R14, —NR14″″—CO—R14, —NR14″″—CS—NR7R14, —NR14″″—CS—OR14, —NR14″″—C(═NR14′″)—NR7R14, —O—CS—NR7R14, —CO—NR7R14, —NR7R14, —CH2—SR4″″, —O—R14′, —OCR14′R14″R14′″, —OCH2—R14′, —O—(CH2)m—R14′, —OCH2—CR14′R14″R14′″, —O—(CH2)m—CR14′R14″R14′″, —O—CR19R20—CR14′R14″R14′″, —OCH2—OR14′, —O—(CH2)p—OR14′, —OCH2—OCR14′R14″R14′″, —O—(CH2)p—OCR14′R14″R14′″, —O—CR19R20—OCR14′R14″R14′″, —OCH2—NR14″R14′″, —O—(CH2)p—NR14″R14′″, —O—CR19R20—NR14″R14′″, —OCH2—CO—R14′, —O—(CH2)m—CO—R14′, —OCH2—CO—CR14′R14″R14′″, —O—(CH2)m—CO—CR14′R14″R14′″, —O—CR19R20—CO—CR14′R14″R14′″, —OCH2—O—CO—R14′, —O—(CH2)p—O—CO—R14′, —OCH2—O—CO—CR14′R14″R14′″, —O—(CH2)p—O—CO—CR14′R14″R14′″, —O—CR19R20—O—CO—CR14′R14″R14′″, —OCH2—O—CO—OR14′, —O—(CH2)p—O—CO—OR14′, —OCH2—O—CO—OCR14′R14″R14′″, —O—(CH2)p—O—CO—OCR14′R14″R14′″, —O—CR19R20—O—CO—OCR14′R14″R14′″, —OCH2—NR14′—CO—R14″, —O—(CH2)p—NR14′—CO—R14″, —OCH2—NR14′—CO—CR14″R14′″R14″″, —O—(CH2)p—NR14′—CO—CR14″R14′″R14″″, —O—CR19R20—NR14′—CO—CR14″R14′″R14″″, —OCH2—NR14′—CO—OR14″, —O—(CH2)p—NR14′—CO—OR14″, —OCH2—NR14′—CO—OCR14″R14′″R14″″, —O—(CH2)p—NR14′—CO—OCR14″R14′″R14″″, —O—CR19R20—NR14′—CO—OCR14″R14′″R14″″, —OCH2—CO—OR14′, —OCH2—CO—OCR14′R14″R14′″, —O—(CH2)m—CO—OR14′, —O—(CH2)m—CO—OCR14′R14″R14′″, —O—CR19R20—CO—OCR14′R14″R14′″, —OCH2—CO—NR14″R14′″, —O—(CH2)m—CO—NR14″R14′″, —OCH2—O—CO—NR14″R14′″, —O—(CH2)p—O—CO—NR14″R14′″, —O—CR19R20—O—CO—NR14″R14′″, —OCH2—NR14″—CO—NR14′″R14″″, —O—(CH2)p—NR14″—CO—NR14′″R14″″, —O—CR19R20—NR14″—CO—NR14′″R14″″, —OCH2—NR14″—C(═NR14′)—NR14′″R″″, —O—(CH2)p—NR14″—C(═NR14′)—NR14′″R14″″, —CR14′R14″R14′″, —CH2—R14′, —(CH2)m—R14′, —CH2—CR14′R14″R14′″, —(CH2)m—CR14′R14″R14′″, —CR19R20—CR14′R14″R14′″, —CH2—OR14′, —(CH2)m—OR14′, —CH2—OCR14′R14″R14′″, —(CH2)m—OCR14′R14″R14′″, —CR19R20—OCR14′R14″R14′″, —CH2—NR14″R14′″, —(CH2)m—NR14″R14′″, —CR19R20—NR14″R14′″, —CH2—CO—R14′,—(CH2)m—CO—R14′, —CH2—CO—CR14′R14″R14′″, —(CH2)m—CO—CR14′R14″R14′″, —CR19R20—CO—CR14′R14″R14′″, —CH2—O—CO—R14′, —(CH2)m—O—CO—R14′, —CH2—O—CO—CR14′R14″R14′″, —(CH2)m—O—CO—CR14′R14″R14′″, —CR19R20—O—CO—CR14′R14″R14′″, —CH2—O—CO—OR14′, —(CH2)m—O—CO—OR14′, —CH2O—CO—OCR14′R14″R14′″, —(CH2)m—O—CO—OCR14′R14″R14′″, —CR19R20—O—CO—OCR14′R14″R14′″, —CH2—NR14′—CO—R14″, —(CH2)m—NR14′—CO—R14″, —CH2—NR14′—CO—CR14″R14′″R14″″, —(CH2)m—NR14′—CO—CR14″R14′″R14″″, —CR19R20—NR14′—CO—CR14″R14′″R14″″, —CH2—NR14′—CO—OR14″, —(CH2)m—NR14′—CO—OR14″, —CH2—NR14′—CO—OCR14″R14′″R14″″, —(CH2)m—NR14′—CO—OCR14″R14′″R14″″, —CR19R20—NR14′—CO—OCR14″R14′″R14″″, —CH2—CO—OR14′, —CH2—CO—OCR14′R14″R14′″,—(CH2)m—CO—OR14′, —(CH2)m—CO—OCR14′R14″R14′″, —CR19R20—CO—OCR14′R14″R14′″, —CH2—CO—NR14″R14′″, —(CH2)m—CO—NR14″R14′″, —CR19R20—CO—NR14″R14′″, —CH2—O—CO—NR14″R14′″, —(CH2)m—O—CO—NR14″R14′″, —CR19R20—O—CO—NR14″R14′″, —CH2—NR14″—CO—NR14′″R14″″, —(CH2)m—NR14″—CO—NR14′″R14″″, —CR19R20—NR14″—CO—NR14′″R14″″, —CH2—NR14″—C(═NR14′)—NR14′″R14″″,—(CH2)m—NR14″—C(═NR14′)—NR14″″, —CR19R20—NR14″—C(═NR14′)—NR14′″R14″″, —NR14—CR14′R14″R14′″, —NR14—CH2—R14′, —NR14—(CH2)m—R14′, —NR14—CH2—CR14′R14″R14′″, —NR14—(CH2)m—CR14′R14″R14′″, —NR14—CR19R20—CR14′R14″R14′″, —NR14—CH2—OR14′, —NR14—(CH2)p—OR14′, —NR14—CH2—OCR14′R14″R14′″, —NR14—(CH2)p—OCR14′OR14″R14′″, —NR14—CR19R20—OCR14′R14″R14′″, —NR14—CH2—NR14″R14′″, —NR14—(CH2)p—NR14″R14′″, —NR14—CR19R20—NR14″R14′″, —NR14—CH2—CO—R14′, —NR14—(CH2)m—CO—R14′, —NR14—CH2—CO—CR14′R14″R14′″, —NR14—(CH2)m—CO—CR14′R14″R14′″, —NR14—CR19R20—CO—CR14′R14″R14′″, —NR14—CH2—O—CO—R14′, —NR14—(CH2)p—O—CO—R14′, —NR14—CH2—O—CR14′R14″R14′″, —NR14—(CH2)p—O—CO—CR14′R14″R14′″, —NR14—CR19R20—O—CO—CR14′R14″R14′″, —NR14—CH2—O—CO—OR14′, —NR14—(CH2)p—O—CO—OR14′, —NR14—CH2—O—CO—OCR14′R14″R14′″, —NR14—(CH2)p—O—CO—OCR14′R14″R14′″, —NR14—CR19R20—O—CO—OCR14′R14″R14′″, —NR14—CH2—NR14′—CO—R14″,—NR14—(CH2)p—NR14′—CO—R14″, —NR14—CH2—NR14′—CO—CR14″R14′″R14″″, —NR14—(CH2)p—NR14′—CO—CR14″R14′″R14″″, —NR14—CR19R20—NR14′—CO—CR14″R14′″R14″″, —NR14—CH2—NR14′—CO—OR14″, —NR14—(CH2)p—NR14′—CO—OR14″, —NR14—CH2—NR14′—CO—OCR14″R14′″R14″″, —NR14—(CH2)p—NR14′—CO—OCR14″R14′″R14″″, —NR14—CR19R20—NR14′—CO—OCR14″R14′″R14″″,—NR14—CH2—CO—OR14′, —NR14—CH2—CO—OCR14′R14″R14′″, —NR14—(CH2)m—CO—OR14′, —NR14—(CH2)m—CO—OCR14′R14″R14′″, —NR14—CR19R20—CO—OCR14′R14″R14′″, —NR14—CH2—CO—NR14″R14′″, —NR14—(CH2)m—CO—NR14″R14′″, —NR14—CR19R20—CO—NR14″R14′″, —NR14—CH2—O—CO—NR14″R14′″, —NR14—(CH2)p—O—CO—NR14″R14′″, —NR14—CR19R20—O—CO—NR14″R14′″, —NR14—CH2—NR14″—CO—NR14′″R14″″, —NR14—(CH2)p—NR14″—CO—NR14′″R14″″, —NR14—CR19R20—NR14″—CO—NR14′″R14″″, —NR14—CH2—NR14″—C(═NR14′)—NR14′″R14″″, —NR14—(CH2)p—NR14″—C(═NR14′)—NR14′″R14″″, —NR14—CR19R20—NR14″—C(═NR14′)—NR14′″R14″″, —NR6R7, —(CH2)n—NR6R14, —(CH2)n—OR14, —(CH2)n—R14, —(CH2)n—SR14, —(CH2)n—SO—R14, —(CH2)n—SO2—R14, —(CH2)n—SO3—R14, —(CH2)n—CO—R14, —(CH2)n—COO—R14, —(CH2)n—O—CO—R14, —(CH2)n—NH—CO—R14, —(CH2)n—CO—NR6R14, —(CH2)n—O—CO—OR14, —(CH2)n—NH—CO—OR14, —(CH2)n—O—CO—NR6R14, —(CH2)n—NH—CO—NR6R14, —(CH2)n—NH—CS—NR6R14, —(CH2)n—NH—C(═NH)—NR6R14, —(CH2)n-aryl, —(CH2)n-heteroaryl, —CR19R20—NR6R14, —CR19R20—OR14, —CR19R20R14, —CR19R20—SR14, —CR19R20—SO—R14, —CR19R20—SO2—R14, —CR19R20—SO3—R14, —CR19R20—CO—R14, —CR19R20—COO—R14, —CR19R20—O—CO—R14, —CR19R20—NH—CO—R14, —CR19R20—CO—NR6R14, —CR19R20—O—CO—OR14, —CR19R20—NH—CO—OR14, —CR19R20—O—CO—NR6R14, —CR19R20—NH—CO—NR6R14, —CR19R20—NH—CS—NR6R14, —CR19R20—NH—C(═NH)—NR6R14, —CR19R20-aryl, —CR19R20-heteroaryl, —CH═CH—R18, —CH═CH—R14, —CR18′═CR18″R18′″, —CR14′═CR14″R14′″;
R11 and R16 can form together with Y2 and Y3 a five-membered or six-membered saturated substituted or unsubstituted carbocyclic ring;
R8 and R9 represent together ═O, ═N—OH, ═N—OR4″″, ═N—R4″″, ═N—NH2, ═N—NHR4″″, ═N—NR4′″R4″″,
thus forming a carbonyl group, or an oxime, or a hydrazone, together with the carbon atom Y1, or R8 and R9 form together a carbocyclic or heterocyclic ring;
R10 and R11 if bound to the same carbon atom Y2 represent together ═O, ═N—OH, ═N—OR4″″, ═N—R4″″, ═N—NH2, ═N—NHR4″″, ═N—NR4′″R4″″,
thus forming a carbonyl group, or an oxime, or a hydrazone, together with the carbon atom Y2, or R10 and R11 form together a carbocyclic or heterocyclic ring;
R12 and R13 represent together ═O, ═N—OH, ═N—OR4″″, ═N—R4″″, ═N—NH2, ═N—NHR4″″, ═N—NR4′″R4″″,
thus forming a carbonyl group, or an oxime, or a hydrazone, together with the carbon atom Y4, or R12 and R13 form together a carbocyclic or heterocyclic ring;
R16 and R17 represent together ═O, ═N—OH, ═N—OR4″″, ═N—R4″″, ═N—NH2, ═N—NHR4″″, ═N—NR4′″R4″″,
thus forming a carbonyl group, or an oxime, or a hydrazone, together with the carbon atom Y3, or R16 and R17 form together a carbocyclic or heterocyclic ring;
R14, R14′, R14″, R14′″, and R14″″ are independently of each other selected from —H, substituted or unsubstituted C3-C10-cycloalkyl, substituted or unsubstituted C1-C8-alkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6-heterocyclyl, substituted or unsubstituted C2-C6-alkenyl, substituted or unsubstituted C2-C6-alkinyl, adamantyl, substituted or unsubstituted alkylaryl, —R18, —R18′, —R18″, —R18′″, —R18″″, —R19, —R20, —R21, —R22, —R23, —(CH2)n—O—CO—R18″″, —(CH2)n—NH—CO—R18″″, —(CH2)n—NH—CO—R18″″, —(CH2)n—R18′, —(CH2)n—R18″, —(CH2)n—R18′″, —(CH2)n—SO—R18′″, —(CH2)n—SO2—R18′″, —(CH2)n—CO—R18″″, —(CH2)n—NH—CO—R18″″, —(CH2)n—CO—NH—NH2, —(CH2)n—SR18″, —C≡C—R18″″, —CR18′═CR18″R18″″,
R18, R18′, R18″, R18′″, R18″″, R19-R33 are independently of each other selected from —H, —OH, —OCH3, —OC2H5, —OC3H7, —O-Cyclo-C3H5, —OCH(CH3)2, —OC(CH3)3, —OPh, —OCH2-Ph, —OCPh3, —OR6, —OR7, —SH, —SCH3, —SC2H5, —SC3H7, —S-cyclo-C3H5, —SCH(CH3)2, —SC(CH3)3, —SR6, —SR7, —NO2, —F, —Cl, —Br, —I, —N3, —CN, —OCN, —NCO, —SCN, —NCS, —CHO, —COCH3, —COC2H5, —COC3H7, —CO-cyclo-C3H5, —COCH(CH3)2, —COC(CH3)3, —COOH, —COCN, —COOCH3, —COOC2H5, —COOC3H7, —COO-cyclo-C3H5, —COOCH(CH3)2, —COOC(CH3)3, —COOR6, —COOR7, —OOC—CH3, —OOC—C2H5, —OOC—C3H7, —OOC-cyclo-C3H5, —OOC—CH(CH3)2, —OOC—C(CH3)3, —OOC—R6, —OOC—R7, —CONH2, —CONHCH3, —CONHC2H5, —CONHC3H7, —CONH-cyclo-C3H5, —CONH[CH(CH3)2], —CONH[C(CH3)3], —CON(CH3)2, —CON(C2H5)2, —CON(C3H7)2, —CON(cyclo-C3H5)2, —CON[CH(CH3)2]2, —CON[C(CH3)3]2, —CONR6R7, —NH2, —NHCH3, —NHC2H5, —NHC3H7, —NH-cyclo-C3H5, —NHCH(CH3)2, —NHC(CH3)3, —N(CH3)2, —N(C2H5)2, —N(C3H7)2, —N(cyclo-C3H5)2, —N[CH(CH3)2]2, —N[C(CH3)3]2, —NR6R7, —SOCH3, —SOC2H5, —SOC3H7, —SO-cyclo-C3H5, —SOCH(CH3)2, —SOC(CH3)3, —SO—R6, —SO—R7, —SO2CH3, —SO2C2H5, —SO2C3H7, —SO2-cyclo-C3H5, —SO2CH(CH3)2, —-SO2C(CH3)3, —SO2R6, —SO2R7, —SO3H, —SO3CH3, —SO3C2H5, —SO3C3H7, —SO3-cyclo-C3H5, —SO3CH(CH3)2, —SO3C(CH3)3, —SO3—R6, —SO3—R7, —OCF3, —OC2F5, —O—COOCH3, —O—COOC2H5, —O—COOC3H7, —O—COO-cyclo-C3H5, —O—COOCH(CH3)2, —O—COOC(CH3)3, —O—COOC—R6, —O—COOC—R7, —NH—CO—NH2, —NH—CO—NHCH3, —NH—CO—NHC2H5, —NH—CO—NHC3H7, —NH—CO—NH-cyclo-C3H5, —NH—CO—NH[CH(CH3)2], —NH—CO—NH[C(CH3)3], —NH—CO—N(CH3)2, —NH—CO—N(C2H5)2, —NH—CO—N(C3H7)2, —NH—CO—N(cyclo-C3H5)2, —NH—CO—N[CH(CH3)2]2, —NH—CO—N[C(CH3)3]2, —NH—CO—N(R6)2, —NH—CO—N(R7)2, —NH—CS—NH2, —NH—CS—NHCH3, —NH—CS—NHC2H5, —NH—CS—NHC3H7, —NH—CS—NH-cyclo-C3H5, —NH—CS—NH[CH(CH3)2], —NH—CS—NH[C(CH3)3], —NH—CS—N(CH3)2, —NH—CS—N(C2H5)2, —NH—CS—N(C3H7)2, —NH—CS—N(cyclo-C3H5)2, —NH-CS—N[CH(CH3)2]2, —NH—CS—N[C(CH3)3]2, —NH—CS—N(R6)2, —NH—CS—N(R7)2, —NH—C(═NH)—NH2, —NH—C(═NH)—NHCH3, —NH—C(═NH)—NHC2H5, —NH—C(═NH)—NHC3H7, —NH—C(═NH)—NH-cyclo-C3H5, —NH—C(═NH)—NH[CH(CH3)2], —NH—C(═NH)—NH[C(CH3)3], —NH—C(═NH)—N(CH3)2, —NH—C(═NH)—N(C2H5)2, —NH—C(═NH)—N(C3H7)2, —NH—C(═NH)—N(cyclo-C3H5)2, —NH—C(═NH)—N[CH(CH3)2]2, —NH—C(═NH)—N[C(CH3)3]2, —NH—C(═NH)—N(R6)2, —NH—C(═NH)—N(R7)2, —O—CO—NH2, —O—CO—NHCH3, —O—CO—NHC2H5, —O—CO—NHC3H7, —O—CO—NH-cyclo-C3H5, —O—CO—NH[CH(CH3)2], —O—CO—NH[C(CH3)3], —O—CO—N(CH3)2, —O—CO—N(C2H5)2, —O—CO—N(C3H7)2, —O—CO—N(cyclo-C3H5)2, —O—CO—N[CH(CH3)2]2, —O—CO—N[C(CH3)3]2, —O—CO—N(R6)2, —O—CO—N(R7)2, —O—CO—OCH3, —O—CO—OC2H5, —O—CO—OC3H7, —O—CO—O-cyclo-C3H5, —O—CO—OCH(CH3)2, —O—CO—OC(CH3)3, —O—CO—OR6, —O—CO—OR7, —CH2F, —CHF2, —CF3, —CH2Cl, —CHCl2, —CCl3, —CH2Br, —CHBr2, —CBr3, —CH2I, —CHI2, —Cl3, —CH2—CH2F, —CH2—CHF2, —CH2—CF3, —CH2—CH2Cl, —CH2—CHCl2, —CH2—CCl3, —CH2—CH2Br, —CH2—CHBr2, —CH2—CBr3, —CH2—CH2I, —CH2—CHI2, —CH2—Cl3, —CH2OH, —CH2SH, —CH2NH2, —CH2N(CH3)2, —CH2N(C2H5)2, —C2H4—OH, —C2H4—SH, —C2H4—NH2, —C2H4—N(CH3)2, —C2H4—N(C2H5)2, —CH2OCH3, —CH2SCH3, —CH2OC2H5, —CH2SC2H5, —C2H4—OCH3, —C2H4—SCH3, —C2H4—OC2H5, —C2H4—SC2H5, —CH3, —C2H5, —C3H7, -cyclo-C3H5, —CH(CH3)2, —C(CH3)3, —C4H9, —CH2—CH(CH3)2, —CH(CH3)—C2H5, —C(CH3)3, —C5H11, —C6H13, —C7H15, —C8H17, —C9H19, —C10H21, -Ph, —CH2-Ph, —CH═CH2, —CH═CHF, —CH═CF2, —CF═CF2, —CH═CHCl, —CH═CCl2, —CCl═CCl2, —CH═CHBr, —CH═CBr2, —CBr═CBr2, —CH2—CH═CH2, —C(CH3)═CH2, —CH═CH—CH3, —CH═CH—CH2—OH, —CH═CH—COOH, —CH═CH—COOCH3, —CH═CH—COOC2H5, —CH═CH-Ph, —C(CH3)═CH—CH3, —C2H4—CH═CH2, —CH═C(CH3)2, —C≡CH, —C≡C—CH3, —CH2—C≡CH, —CPh3;
substituted or unsubstituted C1-C8-alkyl, substituted or unsubstituted C2-C6-alkenyl, substituted or unsubstituted C2-C6-alkinyl, substituted or unsubstituted C3-C10-cycloalkyl, substituted or unsubstituted aryl, -Ph, —CH2Ph, substituted phenyl, substituted benzyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted C1-C6-heterocyclyl;
m and n are independently of each other integers from 0-6;
p is an integer from 1-6;
q is an integer from 2-6;
and stereoisomeric and regioisomeric forms and pharmaceutically acceptable salts of these compounds;
under the proviso that if Y1—Y2—Y3—Y4 represent
R2 does not represent —COOR4. In this case, R2 represents preferably —CO—NH2.
The compounds No. 1-287 mentioned in the PCT application WO 03/084947 A1 on pages 9-25 are herewith excluded from the scope of the present invention by disclaimer.
The following compounds are also excluded by disclaimer:
2-(Toluene-4-sulfonylamino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[(5-Bromo-2-hydroxy-benzylidene)-amino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[(2-Chloro-benzylidene)-amino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
Furan-2-carboxylic acid[3-(4-methoxy-phenylcarbamoyl)4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl]-amide;
2-[3-(4-Methoxy-phenyl)-3-(2,2,2-trifluoro-acetylamino)-propionyl-amino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-(7-Ethyl-4-oxo-3-phenyl-3 ,4,5,6,7,8-hexahydro-benzo)[4,5]thieno[2,3-d]pyrimidin-2-ylsulfanyl)-N-(2-isopropoxy-phenyl)-acetamide;
N,N-Diethyl-2-(7-ethyl-4-oxo-3-phenyl-3,4,5,6,7,8-hexahydro-benzo)[4,5]thieno[2,3-d]pyrimidin-2-ylsulfanyl)-acetamide;
2-(2,2,3,3-Tetrafluro-propionylamino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[3-(2,2,2-Trichloro-1-propionylamino-ethyl)-thioureido]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-(2,4-Dichlorophenoxy)-N-(4-oxo-2-propyl-5,6,7,8-tetrahydro-4H-benzo[4,5]thieno[2,3-d]pyrimidin-3-yl)-acetamide;
6-Methyl-2-[(thiophene-2-carbonyl)-amino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[3-(3,4-Dichloro-phenyl-ureido]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-(2-Piperidin-1-yl-acetylamino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[2-(4-Methyl-piperazin-1-yl)-acetylamino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-(3-Furan-2-yl-acryloylamino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[(2-Ethoxy-benzylidene)-amino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
6-Methyl-2-(3-phenyl-propionylamino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
Tetrahydro-furan-2-carboxylic acid(3-carbamoyl-6-methyl-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-amide;
2-(4-Fluoro-benzenesulfonylamino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
6-tert-Butyl-2-[2-(5-methyl-3-trifluormethyl-pyrazol-1-yl)-acetyl-amino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
6-tert-Butyl-2-[2-(3,5-dimethyl-4-nitro-pyrazol-1-yl)-acetylamino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-Amino-6-tert-butyl-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[2-(3,5-Dimethyl-4-nitro-pyrazol-1-yl)-acetylamino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-(3-Carboxy-acryloylamino)-6-(1,1-dimethyl-propyl)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
6-(1,1-Dimethyl-propyl)-2-[(5-methyl-furan-2-carbonyl)-amino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-(Cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
2-[2-(4-Nitrophenyl)-acetylamino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
6-Methyl-2-[2-(3-nitro-[1,2,4]triazol-1-yl)-acetylamino]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide;
Furan-2-carboxylic acid[3-(2-hydroxy-ethylcarbamoyl)-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl]-amide;
2-Acetylamino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-(2,2,-Dimethyl-propionylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-Isobutyrylamino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-(2-Methyl-acryloylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-[(Thiophene-2-carbonyl)-amino]-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
Furan-2-carboxylic acid(3-carbamoyl-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-amide;
2-(Cyclobutanecarbonyl-amino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-(2-Methyl-butyrylamino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-(Cyclopropanecarbonyl-amino)-4-methyl-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
6-tert-Butyl 2-(cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-(Cyclopropanecarbonyl-amino)-6-methyl-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-(Cyclopropylmethyl-amino)-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-(Cyclohexanecarbonyl-amino)4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-Acetylamino-6-methyl-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carboxylic acid amide;
2-Amino-4,7-dihydro-5H-thieno[2,3-c]-thiopyran-3-carboxylic acid amide;
2-(Cyclopropanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]-thiopyran-3-carboxylic acid amide;
2-(Cyclopropanecarbonyl-amino)-6λ4-oxo-4,5,6,7-tetrahydro-thieno[2,3-c]thiopyran-3-carboxylic acid amide.
Not preferred are compounds wherein R1 and R3 are both hydrogen.
The afore-mentioned disclaimer is only directed to the substance claim 1, but not to any one of the use claims 9-37.
Preferred are compounds wherein X1 is S.
Furthermore, compounds are preferred wherein R8 and R9 represent hydrogen.
Also preferred are compounds wherein R1 represents hydrogen. Preferred are compounds wherein the residue Y1-Y2-Y3-Y4 bears at least one further substituent selected from R8-R17 which is different from hydrogen, i.d. Y1 or Y2 or Y3 or Y4 bears a further substituent R8-R17 which is different from hydrogen.
Preferably, —R5′, R10, R11, R12, R13, R16 and R17 represent independently of each other —F, —Cl, —Br, —I, —H, —OH, —OCH3, —OC2H5, —COCH3, and especially preferred —F, —Cl, —Br, —I.
Preferred are compounds wherein R2 represents —CO—NH—R4, and more preferred wherein R2 represents —CO—NH2.
Still preferred are compounds wherein R10 and R11 represent a smaller group, such as —CH2F, —CHF2, —CF3, —CH2OH, —CH2NH2, —CH3, —C2H5, —C3H7, -cyclo-C3H5, —CH(CH3)2, —CH═CH2, —OH, —OCH3, —OC2H5, —OC3H7, —F, —Cl, —H, —COCH3, —COOH, —COOCH3, —CONH2, —NH2, —N(CH3)2, —SOCH3, —SO2CH3, —SO3H, —OCF3.
Preferred are the following substructures (IIa)-(IIf), wherein R2, R3, R8, R9, R10, R11, R12, R13, R16, and R17 have the meanings as defined in claim 1 or any part of the description:
Further preferred are compounds of general formula (IIa) wherein R2 represents —CONH2 and/or wherein R3 represents —CO—R5 or —CO—R5′. Preferred are compounds of formula (IIa), wherein at least one substituent R8-R13 or R16 or R17 is not hydrogen.
Not preferred are compounds of formula (IIa) wherein R8-R17 are hydrogen and R2 represents —CONH2, —CONH—CH3, —CONH—C2H5, and R3 represents —CONH—R*, and —R* represents —CH3, —C2H5, —C3H7, —CH(CH3)2, —C4H9, cyclohexyl, -Ph, chlorophenyl.
Disclaimed are the following compounds:
wherein R5 in formula (A) represents methyl, 1-propyl, n-butyl, cyclohexyl, phenyl, or 4-chlorophenyl;
in formula (B) R5 represents methyl, ethyl, 1-propyl, and R* represents —NH—C2H5, —NH2, —OCH3, —OC2H5, and Y2 represents —CH2—, —O—, —S—, or —NH—;
and in formula (C) R* represents —NH2, —OCH3, and R5 represents methyl or ethyl.
The following compounds are also disclaimed:
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-acetamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-propionamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-2-butynoic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-cyanoacetamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-cyclopropanecarboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-isobutyramide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-3,3-dimethylacrylic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-2-ketobutyramide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-N,N-dimethylglycinamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-3-chloropropionamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-imidazol-4-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-pyrrole-2-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-cyclopentanecarboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-1-cyanocyclopropane-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-N-acetylglycinamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-pyrrole-3-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-benzamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-4-pyrazolecarboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-picolinic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-nicotinic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-isonicotinic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-2-pyrazinecarboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-1-methylpyrrole-2-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-3-methyl-2-furoic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-5-methylisoxazole-4-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-3-methylisoxazole-4-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-thiophene-2-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-thiophene-3-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-dl-pyroglutamic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-1-(aminocarbonyl)-1-cyclopropanecarboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-o-toluic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-5-methylisoxazole-3-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-m-toluic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-3-aminopyrazole-4-carboxamide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-p-toluic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-salicylic amide;
N-[3-carbamoyl-4,5,6,7-tetrahydrobenzo[b]thien-2-yl]-3-hydroxybenzamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-3,4,5-trimethoxybenzamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-2,4,6-trimethoxybenzamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-3-chlorobenzo[b]thiophene-2-carboxamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-3-(phenylsulfonyl)propionamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-4-toluenesulfonylacetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-4-methylsulfonylphenylacetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-5-fluoroindole-3-acetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-3-phthalimido-propionamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-5-methoxy-2-methyl-3-indoleacetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-5-methoxy-1-indanone-3-acetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-5-(4-chlorophenyl)-2-furoic amide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-6-chlorokynurenic amide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-N′-(4-chlorophenyl)maleamic amide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-N′-p-tosylglycinamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-5-chloroindole-2-carboxamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-N′-(1-naphthyl)maleamic amide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-3-iodobenzamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-4-iodobenzamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-N-m-tolylphthalamic amide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-N′-acetyl-dl-histidine;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-3-acetamino-6-bromobenzamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-2-acetamido-5-bromobenzamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-2-iodophenylacetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-4-iodophenylacetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-8-(3-carboxamidopropyl)-1,3-dimethylxanthine;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-7-bromokynurenic amide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-N′-benzoyl-dl-phenylalaninamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-indole-3-butyramide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-4-chloroindole-3-acetamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-dl-desthiobiotin;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-4,6-dichloroindole-2-carboxamide;
N-[3-carbamoyl-6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl]-N′-benzoyl-histidinamid.
Also preferred are the following substructures (IIIa)-(IIIf), wherein R2, R3, R9, R10, R11, R12, R13, and X1 have the meanings as defined in claim 1 or any part of the description:
Especially the compounds of general formula (IIIa) are preferred
wherein
X1 is selected from S, O, NR4′,
R4′ is selected from H, substituted or unsubstituted C1-C6-alkyl,
R2 is selected from —CO—NH—R4, —CS—NH—R4, —SO2—NH—R4;
wherein R4 is selected from —H, HO-substituted, H2N-substituted or HS-substituted C1-C6-alkyl,
R3 is selected from H, —C(═O)R5, —C(═S)R5, —C(═NH)R5 and —SO2R5,
wherein
R5 is selected from substituted or unsubstituted C3-C6-cycloalkyl, C1-C6-alkyl, aryl, heteroaryl, heterocycloalkyl, C2-C4-alkenyl, C2-C4-alkinyl, adamantyl,
or —(CH2)n—NR6R7,
wherein R6 and R7 are independently selected from substituted or unsubstituted C1-C4-alkyl or C2-C4-alkenyl and wherein n=1 to 6,
or NR6R7,
wherein
R6 is selected from H, C1-C6-alkyl, and
R7 is selected from substituted or unsubstituted C3-C6-cycloalkyl, C1-C6-alkyl, aryl, heteroaryl, heterocycloalkyl, C2-C4-alkenyl, C2-C4-alkinyl, or adamantyl,
R8 is H and R9 is selected from H, substituted or unsubstituted C1-C6-alkyl
R10 is selected from H, substituted or unsubstituted C1-C6-alkyl, C1-C6-alkoxy, or OH
R11 is selected from H and substituted or unsubstituted C1-C6-alkyl
R12 is selected from H and substituted or unsubstituted C1-C6-alkyl, C1-C6-alkoxy, or OH, and
R13 is selected from H or substituted or unsubstituted C1-C6-alkyl,
and stereoisomeric and regioisomeric forms and pharmaceutically acceptable salts of these compounds.
In a preferred embodiment of general formula (IIIa) X1 is S.
In a further preferred embodiment of general formula (IIIa) X1 is NR4′, and NR4′ is selected from H, substituted or unsubstituted C1-C6-alkyl, and preferably is methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec.-butyl, iso-butyl, tert.-butyl, or benzyl.
In a further preferred embodiment of general formula (IIIa) X1 is O.
In a further preferred embodiment of general formula (IIIa) R2 is —CO—NH—R4 and R4 is selected from H, HO-substituted, H2N-substituted or HS-substituted C1-C4-alkyl, and preferably is H.
In a further preferred embodiment of general formula (IIIa) R2 is —CS—NH—R4 and R4 is selected from H, HO-substituted, H2N-substituted or HS-substituted C1-C4-alkyl, and preferably is H.
In a further preferred embodiment of general formula (IIIa) R2 is —SO2—NH—R4 and R4 is selected from H, HO-substituted, H2N-substituted or HS-substituted C1-C4-alkyl, and preferably is H.
In yet another preferred embodiment of general formula (IIIa) R4 is selected from the group consisting of —H, —CH2—CH2—OH, —CH2—CH2—NH2, —CH2—CH2—SH, —CH2—CH(OH)—CH3, —CH2—CH(SH)CH3, or —CH2—CH(NH2)—CH3.
In a further preferred embodiment of general formula (IIIa) R3 is —CO—R5, —CS—R5, —CO—R5′, or —CS—R5′, and more preferably —CO—R5 or —CO—R5′ and most preferably —CO—R5.
In a further preferred embodiment of general formula (IIIa) R3 is —SO2—R5 or —SO2—R5′ and more preferably —SO2—R5.
In yet another preferred embodiment of general formula (IIIa) R5 or R5′ is selected from the group consisting of substituted or unsubstituted methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, C1-C6-cycloalkyles substituted by at least one methyl or carboxyl group, phenyl, furanyl, thienyl, pyrrolyl, pyridyl, pyrrolidinyl, piperidinyl, tetrahydrofuranyl, ethenyl, prop-1-enyl, prop-2-enyl, but-1-enyl, but-2-enyl, but-3-enyl, prop-1-inyl, prop-2-inyl, but-1-inyl, but-2-inyl, but-3-inyl, adamantyl, or NR6R7, wherein R6 is H and R7 is selected from substituted or unsubstituted C3-C6-cycloalkyl, C1-C6-alkyl, aryl, heteroaryl, heterocycloalkyl, C2-C4-alkenyl, C2-C4-alkinyl, or adamantyl.
In yet another preferred embodiment of general formula (IIIa) R5 or R5′ is selected from the group consisting of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl-substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, methyl-substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, carboxyl substituted cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, furanyl, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert.-butyl, prop-1-enyl, but-1-enyl, adamantyl, 3,4-difluorophenyl or NR6R7, wherein R6 is H and R7 is selected from substituted or unsubstituted C3-C6-cycloalkyl, C1-C6-alkyl, aryl, heteroaryl, heterocycloalkyl, C2-C4-alkenyl, C2-C4-alkinyl, or adamantyl, and R7 preferably is phenyl or 3,4-difluorophenyl.
In another preferred embodiment of general formula (IIIa) R7 is selected from substituted or unsubstituted C3-C6-cycloalkyl, C1-C6-alkyl, heteroaryl, heterocycloalkyl, C2-C4-alkenyl, C2-C4-alkinyl, or adamantyl. In a further embodiment of general formula (IIIa), the compound 5,5-dimethyl-2-(3-phenyl-ureido)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide is excluded from the compounds according to the present invention.
In yet another embodiment of general formula (IIIa) R7 is selected from substituted or unsubstituted C3-C6-cycloalkyl, C1-C6-alkyl, aryl, heteroaryl, heterocycloalkyl, C2-C4-alkenyl, C2-C4-alkinyl, or adamantyl, and R10 is selected from H, substituted or unsubstituted C1-C6-alkoxy, or OH.
In yet another preferred embodiment of general formula (IIIa) R8 is H and R9 is selected from H, or substituted or unsubstituted C1-C6-alkyl.
In a further preferred embodiment of general formula (IIIa) R10, R11, R12, and R13 are independently selected from H and substituted or unsubstituted C1-C6-alkyl, and preferably from H or methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl or tert.-butyl.
In yet another preferred embodiment of general formula (IIIa) R10 and R11 are methyl and R12 and R13 are H, or R10, R11, R12, and R13 are H, or R10, R11, R12, and R13 are methyl, or R10 and R11 are H and R12 and R13 are methyl.
In yet another preferred embodiment of general formula (IIIa) R10 is selected from substituted or unsubstituted C1-C6-alkoxy or OH and R11 is selected from H or substituted or unsubstituted C1-C6-alkyl.
In yet another preferred embodiment of general formula (IIIa) R12 is selected from substituted or unsubstituted C1-C6-alkoxy or OH and R13 is selected from H or substituted or unsubstituted C1-C6-alkyl.
In a further preferred embodiment of general formula (IIIa) R4′ and/or R1 is selected from the group consisting of methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert.-butyl or benzyl.
In a further preferred embodiment of general formula (IIIa) R6 and R7 are independently selected from methyl, ethyl and propyl or allyl, and preferably are methyl.
Preferred are the following substructures (IVa)-(IVf) of general formula (I), wherein R2, R3, R8, R9, R10, R11, R12, R13, R15, and X1 have the meanings as defined in claim 1 or any part of the description:
Furthermore, preferred are the following substructures (Va)-(Vd), wherein R2, R3, R10, R11, R12, and X1 have the meanings as defined in claim 1 or any part of the description:
Still preferred are the following substructures (VIa)-(VId), wherein R2, R3, R10, R11, R12, R13, and X1 have the meanings as defined in claim 1 or any part of the description:
Excluded from the scope of the present invention via disclaimer are the following compounds:
2-Amino-6-(4-fluoro-phenyl)-benzo[b]thiophene-3-carboxylic acid amide;
2-Ureido-benzo[b]thiophene-3-carboxylic acid amide.
Not preferred are compounds of the general formula (VIc), wherein R3 represents —CO—NH2. Also not preferred are compounds of the general formula (VIc), wherein R3 represents —CO—NH2 and R10, R11, R13 are hydrogen and R16 represents —H, —CN, —CF3, halogen, aryl, heteroaryl, alkyl, O-alkyl, or S-alkyl.
Preferred are still the following substructures (VIIa)-(VIId), wherein R1, R2, R3, R8, R10, R17, and X1 have the meanings as defined in claim 1 or any part of the description:
Still preferred are the following substructures (VIIIa)-(VIIId), wherein R1, R2, R3, R10, R17, and X1 have the meanings as defined in claim 1 or any part of the description:
Preferred are still the following substructures (IXa)-(IXd), wherein R1, R2, R3, R10, R12, and X1 have the meanings as defined in claim 1 or any part of the description:
Preferred are still the following substructures (Xa)-(Xd), wherein R1, R2, R3, R8, R9, R10, R11, R12, R13, and X1 have the meanings as defined in claim 1 or any part of the description:
Preferred are still the following substructures (XIa)-(XId), wherein R1, R2, R3, R8, R9, R10, R11, R12, R13, R15, R16, R17, and X1 have the meanings as defined in claim 1 or any part of the description:
Preferred are especially all compounds of any of the above-mentioned general formula (I)-(XI), wherein at least one of the substituents R3-R13, R15-R17 are different from hydrogen, more preferably wherein at least one substituent R12, R13, R16, or R17 is different from hydrogen. Still further preferred are compounds wherein R12, R13 and/or R16 are residues containing at least one oxygen and/or nitrogen atom such as the residues: —OOC—O—R14′, —O—CO—R14′, —COO—R14′, —O—CO—NR7R14, —CO—NR7R14, —NR7R14, —O—R14′, —OCR14′R14″R14′″, —OCH2—R14′, —O—(CH2)m—R14′, —OCH2—CR14′R14″R14′″, —O—(CH2)m—CR14′R14″R14′″, —O—(CH2)p—OCR14′R14″R14′″, —O—(CH2)p—NR14″R14′″, —O—(CH2)m—CO—R14′, —O—(CH2)p—O—CO—R14′, —O—(CH2)p—O—CO—OR14′, —O—(CH2)p—NR14′—CO—R14″, —O—(CH2)p—NR14′—CO—OR14″, —O—(CH2)m—CO—OCR14′R14″R14′″, —O—CR19R20—CO—OCR14′R14″R14′″, —O—(CH2)m—CO—NR14″R14′″, —O—(CH2)p—O—CO—NR14″R14′″, —O—CR19R20—O—CO—NR14″R14′″, —(CH2)m—OR14′, —(CH2)m—NR14″R14′″, —(CH2)m—O—CO—R14′, —(CH2)m—NR14′—CO—R14″, —(CH2)m—NR14′—CO—OR14″, —(CH2)m—CO—OR14′, —(CH2)m—CO—NR14″R14′″, —(CH2)m—O—CO—NR14″R14′″.
As used herein, the term “unsubstituted C1-C8-alkyl” refers to —CH3, —C2H5, —C3H7, —CH(CH3)2, —C4H9, —CH2—CH(CH3)2, —CH(CH3)—C2H5, —C(CH3)3, —C5H11, —CH(CH3)—C3H7, —CH2—CH(CH3)—C2H5, —CH(CH3)—CH(CH3)2, —C(CH3)2—C2H5, —CH2—C(CH3)3, —CH(C2H5)2, —C2H4—CH(CH3)2, —C6H13, —C3H6—CH(CH3)2, —C2H4—CH(CH3)—C2H5, —CH(CH3)—C4H9, —CH2—CH(CH3)—C3H7, —CH(CH3)—CH2—CH(CH3)2, —CH(CH3)—CH(CH3)—C2H5, —CH2—CH(CH3)—CH(CH3)2, —CH2—C(CH3)2—C2H5, —C(CH3)2—C3H7, —C(CH3)2—CH(CH3)2, —C2H4—C(CH3)3, —CH(CH3)—C(CH3)3, —C7H15, —C8H17, —C4H8—CH(CH3)2, —C5H10—CH(CH3)2. Consequently, C1-C7-alkyl will refer the the residues disclosed before having 1 to 7 carbon atoms and C1-C3-alkyl will refer to the residues mentioned before having 1 to 3 carbon atoms.
Preferred are —CH3, —C2H5, —C3H7, —CH(CH3)2, —C4H9, —CH2—CH(CH3)2, —CH(CH3)—C2H5, —C(CH3)3, and —C5H11. Especially preferred are —CH3, —C2H5, —C3H7, and —CH(CH3)2.
The term “unsubstituted C2-C6-alkenyl” refers to —CH═CH2, —CH2—CH═CH2, —C(CH3)═CH2, —CH═CH—CH3, —C2H4—CH═CH2, —CH2—CH═CH—CH3, —CH═CH—C2H5, —CH2—C(CH3)═CH2, —CH(CH3)—CH═CH, —CH═C(CH3)2, —C(CH3)═CH—CH3, —CH═CH—CH═CH2, —C3H6—CH═CH2, —C2H4—CH═CH—CH3, —CH2—CH═CH—C2H5, —CH═CH—C3H7, —CH2—CH═CH—CH═CH2, —CH═CH—CH═CH—CH3, —CH═CH—CH2—CH═CH2, —C(CH3)═CH—CH═CH2, —CH═C(CH3)—CH═CH2, —CH═CH—C(CH3)═CH2, —C2H4—C(CH3)═CH2, —CH2—CH(CH3)—CH═CH2, —CH(CH3)—CH2—CH═CH2, —CH2—CH═C(CH3)2, —CH2—C(CH3)═CH—CH3, —CH(CH3)—CH═CH—CH3, —CH═CH—CH(CH3)2, —CH═C(CH3)—C2H5, —C(CH3)═CH—C2H5, —C(CH3)═C(CH3)2, —C(CH3)2—CH═CH2, —CH(CH3)—C(CH3)═CH2, —C(CH3)═CH—CH═CH2, —CH═C(CH3)—CH═CH2, —CH═CH—C(CH3)═CH2, —C4H8—CH═CH2, —C3H6—CH═CH—CH3, —C2H4—CH═CH—C2H5, —CH2—CH═CH—C3H7, —CH═CH—C4H9, —C3H6—C(CH3)═CH2, —C2H4—CH(CH3)—CH═CH2, —CH2—CH(CH3)—CH2—CH═CH2, —CH(CH3)—C2H4—CH═CH2, —C2H4—CH═C(CH3)2, —C2H4—C(CH3)═CH—CH3, —CH2—CH(CH3)—CH═CH—CH3, —CH(CH3)—CH2—CH═CH—CH3, —CH2—CH═CH—CH(CH3)2, —CH2—CH═C(CH3)—C2H5, —CH2—C(CH3)═CH—C2H5, —CH(CH3)—CH═CH—C2H5, —CH═CH—CH2—CH(CH3)2, —CH═CH—CH(CH3)—C2H5, —CH═C(CH3)—C3H7, —C(CH3)═CH—C3H7, —CH2—CH(CH3)—C(CH3)═CH2, —CH(CH3)—CH2—C(CH3)═CH2, —CH(CH3)—CH(CH3)—CH═CH2, —CH2—C(CH3)2—CH═CH2, —C(CH3)2—CH2—CH═CH2, —CH2—C(CH3)═C(CH3)2, —CH(CH3)—CH═C(CH3)2, —C(CH3)2—CH═CH—CH3, —CH(CH3)—C(CH3)═CH—CH3, —CH═C(CH3)—CH(CH3)2, —C(CH3)═CH—CH(CH3)2, —C(CH3)═C(CH3)—C2H5, —CH═CH—C(CH3)3, —C(CH3)2—C(CH3)═CH2, —CH(C2H5)—C(CH3)═CH2, —C(CH3)(C2H5)—CH═CH2, —CH(CH3)—C(C2H5)═CH2, —CH2—C(C3H7)═CH2, —CH2—C(C2H5)═CH—CH3, —CH(C2H5)—CH═CH—CH3, —C(C4H9)═CH2, —C(C3H7)═CH—CH3, —C(C2H5)═CH—C2H5, —C(C2H5)═C(CH3)2, —C[C(CH3)3]═CH2, —C[CH(CH3)(C2H5)]═CH2, —C[CH2—CH(CH3)2]═CH2, —C2H4—CH═CH—CH═CH2, —CH2—CH═CH—CH2—CH═CH2, —CH═CH—C2H4—CH═CH2, —CH2—CH═CH—CH═CH—CH3, —CH═CH—CH2—CH═CH—CH3, —CH═CH—CH═CH—C2H5, —CH2—CH═CH—C(CH3)═CH2, —CH2—CH═C(CH3)—CH═CH2, —CH2—C(CH3)═CH—CH═CH2, —CH(CH3)—CH═CH—CH═CH2, —CH═CH—CH2—C(CH3)═CH2, —CH═CH—CH(CH3)—CH═CH2, —CH═C(CH3)—CH2—CH═CH2, —C(CH3)═CH—CH2CH═CH2, —CH═CH—CH═C(CH3)2, —CH═CH—C(CH3)═CH—CH3, —CH═C(CH3)—CH═CH—CH3, —C(CH3)═CH—CH═CH—CH3, —CH═C(CH3)—C(CH3)═CH2, —C(CH3)═CH—C(CH3)═CH2, —C(CH3)═C(CH3)—CH═CH2, and —CH═CH—CH═CH—CH═CH2.
Preferred are —CH═CH2, —CH2—CH═CH2, —C(CH3)═CH2, —CH═CH—CH3, —C2H4—CH═CH2, —CH2—CH═CH—CH3. Especially preferred are —CH═CH2, —CH2—CH═CH2, and —CH═CH—CH3.
The term “unsubstituted C2-C6-alkynyl” refers to —C≡CH, —C≡C—CH3, —CH2—C≡CH, —C2H4—C≡CH, —CH2—C≡C—CH3, —C≡C—C2H5, —C3H6—C≡CH, —C2H4—C≡C—CH3, —CH2—C≡C—C2H5, —C≡C—C3H7, —CH(CH3)—C≡CH, —CH2—CH(CH3)—C≡CH, —CH(CH3)—CH2—C≡CH, —CH(CH3)—C≡C—CH3, —C4H8—C≡CH, —C3H6—C≡C—CH3, —C2H4—C≡C—C2H5, —CH2—C≡C—C3H7, —C≡C—C4H9, —C2H4—CH(CH3)—C≡CH, —CH2—CH(CH3)—CH2—C≡CH, —CH(CH3)—C2H4—C≡CH, —CH2—CH(CH3)—C≡C—CH3, —CH(CH3)—CH2—C≡C—CH3, —CH(CH3)—C≡C—C2H5, —CH2—C≡C—CH(CH3)2, —C≡C—CH(CH3)—C2H5, —C≡C—CH2—CH(CH3)2, —C≡C—C(CH3)3, —CH(C2H5)—C≡C—CH3, —C(CH3)2—C≡C—CH3, —CH(C2H5)—CH2—C≡CH, —CH2—CH(C2H5)—C≡CH, —C(CH3)2—CH2—C≡CH, —CH2—C(CH3)2—C≡CH, —CH(CH3)—CH(CH3)—C≡CH, —CH(C3H7)—C≡CH, —C(CH3)(C2H5)—C≡CH, —C≡C—C≡CH, —CH2—C≡C—C≡CH, —C≡C—C≡C—CH3, —CH(C≡CH)2, —C2H4—C≡C—C≡CH, —CH2—C≡C—CH2—C≡CH, —C≡C—C2H4—C≡CH, —CH2—C≡C—C≡C—CH3, —C≡C—CH2—C≡C—CH3, —C≡C—C≡C—C2H5, —C≡C—CH(CH3)—C≡CH, —CH(CH3)—C≡C—C≡CH, —CH(C≡CH)—CH2—C≡CH, —C(C≡CH)2—CH3, —CH2—CH(C≡CH)2, —CH(C≡CH)—C≡C—CH3, —C≡C—CH≡CH2, —CH≡CH—C≡CH, —CH2—C≡C—CH═CH2, —CH2—CH═CH—C≡CH, —C≡C—CH═CH—CH3, —CH═CH—C≡C—CH3, —C≡C—CH2—CH═CH2, —CH═CH—CH2—C≡CH, —C≡C—CH2—C≡CH, —C(CH3)═CH—C≡CH, —CH═C(CH3)—C≡CH, —C≡C—C(CH3)═CH2, and —C≡C—C≡C—C≡CH.
Preferred are —C≡CH, —C≡C—CH3.
As used herein, the terms “substituted C1-C8-alkyl”, “substituted C2-C6-alkenyl”, and “substituted C2-C6-alkynyl” refer to the above-mentioned “unsubstituted C1-C8-alkyl”, “unsubstituted C2-C6-alkenyl”, and “unsubstituted C2-C6-alkynyl” residues which may be substituted with one, two, three, four, five, six, or seven substituents independently selected from the group referred to as R19-R33. Preferred are the following substituents: —OH, —OCH3, —OC2H5, —OC3H7, —O-cyclo-C3H5, —OCH(CH3)2, —OC(CH3)3, —OPh, —OCH2-Ph, —OCPh3, —OR6, —OR7, —SH, —SCH3, —SC2H5, —NO2, —F, —Cl, —Br, —I, —COCH3, —COC2H5, —COC3H7, —CO-cyclo-C3H5, —COCH(CH3)2, —COC(CH3)3, —COOH, —COOCH3, —COOC2H5, —COOC3H7, —OOC—CH3, —OOC—C2H5, —OOC—C3H7, —CONH2, —CON(CH3)2, —CON(C2H5)2, —NH2, —NHCH3, —NHC2H5, —NHC3H7, —NH-cyclo-C3H5, —NHCH(CH3)2, —NHC(CH3)3, —N(CH3)2, —N(C2H5)2, —N(C3H7)2, —SOCH3, —SOC2H5, —SO2CH3, —SO2C2H5, —SO3H, —SO3CH3, —SO3C2H5, —OCF3, —O—COOCH3, —O—COOC2H5, —O—COOC3H7, —O—CO—NH2, —O—CO—N(CH3)2, —O—CO—N(C2H5)2, —CH2F, —CF3, —CH2Cl, —CCl3, —CH2Br, —CH2I, —CH2OH, —CH2SH, —CH2NH2, —CH2N(CH3)2, —CH2N(C2H5)2, —C2H4—OH, —C2H4—SH, —C2H4—NH2, —C2H4—N(CH3)2, —C2H4—N(C2H5)2, —CH2OCH3, —CH2SCH3, —CH2OC2H5, —C2H4—OCH3, —C2H4—OC2H5, —CH3, —C2H5, —C3H7, -cyclo-C35, —CH(CH3)2, —C(CH3)3, —C4H9, —CH2—CH(CH3)2, —CH(CH3)—s C2H5, —C(CH3)3, —C5H11, —C6H13, —C7H15, —C8H17, —C9H19, —C10H21, -Ph, —CH2-Ph, —CH═CH2, —CH═CCl2, —CH2—CH═CH2, —C(CH3)═CH2, —CH═CH—CH3, —CH═CH—CH2—OH, —CH═CH—COOH, —CH═CH—COOCH3, —CH═CH—COOC2H5, —C(CH3)═CH—CH3, —C2H4—CH═CH2, —CH═C(CH3)2, —C≡CH, —C≡C—CH3, —CH2—C≡CH.
As used herein, the term “unsubstituted C3-C10-cycloalkyl” refers to
The term “substituted C3-C10-cycloalkyl” refers to the above-mentioned carbocyclic residues which are substituted with one, two, three, four, five, six, or seven substituents independently selected from the group referred to as R19-R33. Preferred substituents are listed above and are the same as summarized for the “substituted C1-C8-alkyl”, “substituted C2-C6-alkenyl”, and “substituted C2-C6-alkynyl” residues.
As used herein, the term “—O—C3-C10-cycloalkyl” refers to a substituted or unsubstituted C3-C10-cycloalkyl residue bound via an oxygen to the bicyclic scaffold. For example, “—O—C3-cycloalkyl” refers to —O-cyclo-C3H5.
Accordingly, the term “—O—C1-C6-heterocyclyl” refers to a substituted or unsubstituted heterocyclic ring containing at least one carbon atom. Said heterocyclic ring is bound via said at least one carbon atom to the bicyclic scaffold through an oxygen linker. Examples for “—O—C4-heterocyclyl” are:
As used herein, the term “substituted phenyl” refers to a phenyl ring substituted with one, two, three, four, or five substituents independently selected from the group referred to as R19-R33.
As used herein, the term “substituted benzyl” refers to the residue —CH2-Ph wherein Ph represents a substituted phenyl as defined above.
As used herein, the term “unsubstituted aryl” refers to phenyl, indenyl, indanyl, naphthyl, 1,2-dihydro-naphthyl, 2,3-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl (tetralinyl), fluorenyl, anthryl(anthracenyl), 9,10-dihydroanthryl, 1,2,3,4-tetrahydro-anthryl, 1,2,3,4,5,6,7,8-octahydro-anthryl, azulenyl, diphenylmethyl, benzyl, triphenylmethyl(trityl), styryl, naphthoquinonyl, acenaphthyl, anthraquinonyl, phenanthryl(phenanthrenyl).
As used herein, the term “substituted aryl” refers to any one of the residues “unsubstituted aryl” substituted with one, two, three, four, five, six, or seven substituents independently selected from the group referred to as R19-R33.
The term “alkylaryl” refers to a substituted or unsubstituted aryl moieties linked to the rest of the molecule via a carbon chain. In its easiest form, “alkylaryl” refers to benzyl. Other examples are styryl, phenylethyl, phenylpropyl.
As used herein, the term “substituted C1-C6-alkoxy” or “substituted C1-C6-alkyloxy” refers to substituted or unsubstituted C1-C6-alkyl, wherein an additional oxygen is present at a non-terminal position. Examples for C1-C6-alkoxy are: —O—CH3, —CH2—O—CH3, —C2H4—O—CH(CH3)2, —CH2—O—C2H5, —CH2—O—CH2—C(CH3)3.
As used herein, the term “unsubstituted C1-C8-acyl” refers to the residues —CO—C1-C7-alkyl, wherein C1-C7-alkyl has the meanings as defined above. Accordingly, the term “substituted C1-C8-acyl” refers to the residues referred to as “unsubstituted C1-C8-acyl” which were substituted with one, two, three, four, five, six, or seven substituents independently selected from the group referred to as R19-R33.
As used herein, the term “unsubstituted C1-C6-heterocyclyl” or “unsubstituted C1-C6-heterocyclyl” refers to carbocycles having at least one heteroatom in the ring such as oxygen, nitrogen, or sulfur. Such heterocycles may be saturated or partially unsaturated but not aromatic. Examples for heterocyclic residues are 1,3-dioxolane, benzo[1,3]dioxolyl, pyrazolinyl, pyranyl, thiomorpholinyl, pyrazolidinyl, piperidyl, piperazinyl, 1,4-dioxanyl, imidazolinyl, pyrrolinyl, imidazolidinyl, morpholinyl, 1,4-dithianyl, pyrrolidinyl, oxozolinyl, oxazolidinyl, isoxazolinyl, isoxazolidinyl, thiazolinyl, thiazolidinyl, isothiazolinyl, isothiazolidinyl, dihydropyranl. The term “C4-heterocyclyl” refers to heterocyclic residues with 4 carbon atoms and at least one heteroatom such as tetrahydrofuran, optionally substituted or unsubstituted. Examples for “C1-heterocyclyl” are diaziridine and oxaziridine.
As used herein, the term “substituted C1-C6-heterocyclyl” or “substituted C1-C6-heterocyclyl” refers to the afore-mentioned heterocycles having one, two, three, four, five, six, or seven substituents independently selected from the group referred to as R19-R33.
As used herein, the term “unsubstituted heteroaryl” refers to heteroaromatic groups which have from 4 to 9 ring atoms, from 1 to 4 of which are selected from O, N and/or S. Preferred groups have 1 or 2 heteroatoms in a 5- or 6-membered aromatic ring. Mono and bicyclic ring systems are included. Typical heteroaryl groups include pyridyl, furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, pyridazinyl, pyrimidyl, pyrazinyl, 1,3,5-triazinyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, indolizinyl, indolyl, isoindolyl, benzo[b]furyl, benzo[b]thienyl, indazolyl, benzimidazolyl, benzthiazolyl, purinyl, quinolizinyl, quinolyl, isoquinolyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, tetrahydroquinolyl, benzooxazolyl, chrom-2-onyl, indazolyl, and the like.
As used herein, the term “substituted heteroaryl” refers to heteroaromatic groups as disclosed before having one, two, three, four, or five substituents independently selected from the group referred to as R19-R33.
As used in the present invention, the term substituted or unsubstituted C1-C6-alkyl or C1-C4-alkyl or C1-C3-alkyl is preferably meant to include linear or branched alkyl groups in which optionally one, two or three of the hydrogen atoms bonded to the carbon chain are substituted by a halogen atom such as F, Cl, Br, or I, preferably F or Cl, a —OH or —SH group, a —NH2 group, methoxy or ethoxy group, or phenyl group. These terms therefore especially comprise, depending on the number of carbon atoms in each respective term, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert.-butyl, —C5H11, —CH2—C(CH3)3, —CH(CH3)—C3H7, —CH2—CH(CH3)—C2H5, —CH(CH3)—CH(CH3)2, —C(CH3)2—C2H5, —CH2—C(CH3)3, —C2H4—CH(CH3)2, —C6H13, —C3H6—CH(CH3)2, —C2H4—CH(CH3)—C2H5, —CH(CH3)—C4H9, —CH2—CH(CH3)—C3H7, —CH(CH3)—CH2—CH(CH3)2, —CH(CH3)—CH(CH3)—C2H5, —CH2—CH(CH3)—CH(CH3)2, —CH2—C(CH3)2—C2H5, —C(CH3)2—C3H7, —C(CH3)2—CH(CH3)2, —C2H4—C(CH3)3, —CH(CH3)—C(CH3)3, optionally substituted in the above described manner, especially to give phenyl substituted alkyles such as benzyl.
Similarly, the term substituted or unsubstituted C3-C6-cycloalkyl is preferably meant to include cyclolalkanes in which optionally one, two or three of the hydrogen atoms bonded to the carbon atoms of the cycle are substituted by a halogen atom such as F, Cl , Br, or I, preferably F or Cl, a —OH or —SH group, a —NH2, methoxy or ethoxy or methyl, ethyl or phenyl group. This term therefore preferably includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl as well as methyl substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, ethyl substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or phenyl substituted cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, optionally substituted in the above described manner.
Similarly, the term unsubstituted or substituted C2-C4-alkenyl is preferably meant to include branched or linear alkenyles in which optionally one, two, three or four of the hydrogen atoms bonded to the carbon atoms of the alkyl are substituted by a halogen atom such as F, Cl , Br, or I, preferably F or Cl. These terms therefore are meant to preferably comprise ethenyl, cis-prop-1-enyl, trans-prop-1-enyl, cis-prop-2-enyl, trans-prop-2-enyl, but-1-enyl, cis-but-2-enyl, trans-but-2-enyl, but-3-enyl, optionally substituted in the above described manner.
Similarly, the term unsubstituted or substituted C2-C4-alkinyl is preferably meant to include branced or linear alkinyles in which optionally one, two, three or four of the hydrogen atoms bonded to the carbon atoms of the alkyl are substituted by a halogen atom such as F, Cl , Br, or I, preferably F or Cl. These terms therefore are meant to preferably comprise prop-1-inyl, prop-2-inyl, but-1-inyl, but-2-inyl, and but-3-inyl, optionally substituted in the described above manner.
The term substituted or unsubstituted aryl is preferably meant to include aromatic compounds, in which one, two or three of the hydrogen atoms bonded to the aromatic ring are substituted by an halogen, such as F, Cl, Br or I, preferably F and Cl, or substituted by —NO2, —OH, —SH, —NH2, —CN, methyl or methoxy. This term is therefore meant to preferably comprise phenyl, 2,3-halogen substituted phenyl, 3,4-halogen substituted phenyl.
The term substituted or unsubstituted heteroaryl is preferably meant to include aromatic groups in which the aromatic ring comprises at least one heteroatom selected from the group N, O, or S, and in which one, two or three of the hydrogen atoms bonded to the aromatic ring are optionally substituted by an halogen, such as F, Cl, Br or I, preferably F and Cl, or substituted by —NO2, —OH, —SH, methyl or methoxy. This term therefore includes preferably furanyl, pyrrolyl, thienyl, and pyridinyl which optionally can be substituted in the above described manner.
The term substituted or unsubstituted heterocycloalkyl is preferably meant to include cycloalkyles in which at least one of the carbon atoms of the ring, preferably 1 or 2 atoms, have been substituted by a heteroatom selected from the group consisting of N, O, and S which optionally and in which one, two or three of the hydrogen atoms bonded to the ring are substituted by an halogen, such as F, Cl, Br or I, preferably F and Cl, or substituted by methyl or methoxy. This term therefore includes preferably pyrrolidinyl, piperidinyl and tetrahydrofuranyl, which optionally can be substituted in the above described manner.
In yet another preferred embodiment of the invention compound according to formula (I) is selected from the group comprising:
The present invention also comprises pharmaceutically active salts of these compounds, all stereoisomeric forms and regioisomeric forms of these compounds or prodrugs thereof.
Other aspects of the present invention relate to the heterobicyclic compounds disclosed herein, for instance 4,7-dihydro-5H-thieno[2,3-c]pyran derivatives or 4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amides as outlined above in the general formula (I), for use as new pharmaceutically active agents, particularly for the prophylaxis and/or treatment of prion diseases, immunological diseases, autoimmune diseases, bipolar and clinical disorders, cardiovascular diseases, cell proliferative diseases, diabetes, inflammation, transplant rejections, erectile dysfunction, neurodegenerative diseases and stroke virally or bacterially induced diseases or infections, especially infections induced by bacteria of the genus legionella, and especially legionnaires disease, or mycobateria-induced infections (including opportunistic infections) and diseases, especially mycobacteria induced meningitis, tuberculosis and leprosy, pharmaceutical compositions comprising these heterobicyclic compounds as active ingredients and methods for treating prion diseases, immunological diseases, autoimmune diseases, bipolar and clinical disorders, cardiovascular diseases, cell proliferative diseases, diabetes, inflammation, transplant rejections, erectile dysfunction, neurodegenerative diseases, stroke, virally and/or bacterially induced diseases, particularly mycobacteria-induced infections, in mammals, including humans, especially for the treatment of treatment of virally or bacterially induced diseases or infections, especially infections induced by bacteria of the genus legionella, and especially legionnaires disease, or mycobateria-induced infections (including opportunistic infections) and diseases, especially mycobacteria induced meningitis, tuberculosis and leprosy.
Surprisingly, it was found that the compounds according to general formula (I) as well as pharmaceutically acceptable salts of these compounds are effective against prion diseases, immunological diseases, autoimmune diseases, bipolar and clinical disorders, cardiovascular diseases, cell proliferative diseases, diabetes, inflammation, transplant rejections, erectile dysfunction, neurodegenerative diseases, stroke, virally and/or bacterially induced diseases, especially mycobacteria-induced infections and diseases at pharmaceutically acceptable concentrations while exhibiting enhanced metabolitic stability. It shall be stressed that the compounds which are excluded from claim 1 by disclaimer are herewith explicitly claimed for any pharmaceutical use thereof.
Additionally, the present invention relates to the use of the compounds of the present invention for the manufacturing of a pharmaceutical composition for the prophylaxis and/or treatment of prion diseases, immunological diseases, autoimmune diseases, bipolar and clinical disorders, cardiovascular diseases, cell proliferative diseases, diabetes, inflammation, transplant rejections, erectile dysfunction, neurodegenerative diseases, stroke, virally and/or bacterially induced diseases, particularly those infections and diseases mentioned above.
The compounds of the present invention are effective against mycobacteria induced infections, particularly tuberculosis, but also e.g. leprosy and mycobacteria-induced meningitis. Mycobacteria which induce or cause these infectious diseases are members of the group comprising the tuberculous bacteria Mycobacterium tuberculosis, M. bovis, M. africanum and M. leprae as well as the non-tuberculous bacteria M. abscessus, M. avium, M. celatum, M. chelonae, M. fortuitum, M. genavense, M. gordonae, M. haemophilum, M. intracellulare, M. kansii, M. malmoense, M. marinum, M. scrofulaceum, M. simiae, M. szulgai, M. ulcerans and M. xenopi. Because of the outstanding clinical importance of tuberculosis, microbiologists have distinguished the so-called “Mycobacterium tuberculosis complex” consisting of Mycobacterium tuberculosis, M. bovis, and M. africanum from all other mycobacteria which form the group of the so-called “atypical mycobacteria” or “non-tuberculous mycobacteria (NTM)”.
The use of 4,5,6,7-tetrahydrobenzo[b]thiophene derivatives in the treatment of mycobacterial infections such as tuberculosis is described in the PCT patent application PCT/EP03/03697. The compounds described therein have been found to be effective in blocking the activity of mycobacterial protein serine/threonine kinases, particularly protein kinase G (PknG), which have been identified as an essential component involved in the persistence and enhanced survival of pathogenic mycobacterial within a macrophage cell line, and thereby provide a mode for the elimination of mycobacteria.
Mycobacterial protein serine/threonine kinases such as PknF, PknI, PknJ, PknL, and particularly protein kinase G (PknG), have been identified as an essential component involved in the persistence and enhanced survival of pathogenic mycobacteria within a macrophage cell line. Furthermore, it could be demonstrated that the activity of PknG is an essential factor for virulence of mycobacteria. In accordance with the present invention, compounds have been found which are blocking the activity of PknG in a submicromolar range thus showing that PknG, PknF, PknI, PknJ, and PknL are suitable targets for recognising diseases, monitoring diseases, and controlling therapy of diseases related to mycobacterial infections. These compounds (inhibitors) were able to induce efficient degradation of mycobacteria within host cells so that the present invention provides a novel mode for elimination of mycobacteria.
By means of an alkaline phosphatase secretion assay for PknG for different PhoA fusion constructs, it could be shown that PknG is a secretory protein that is secreted outside the mycobacterial cells. The secreted PknG can phosphorylate host cell proteins that might be critical in survival of mycobacterium in host cells.
Additionally, biologically active 4,7-dihydro-5-H-thieno[2,3-c]pyran and 4,7-dihydro-5-H-thieno[2,3-c]thiopyran derivatives are described in Biorg. Med. Chem. Letters 2002, 12, 1897-1900, in which compounds which inhibit TNF-alpha-production are described, in J. Med. Chem. 2002, 45, 4443-4459, in which compounds are described which act as protein-tyrosine phosphatase 1B (PTP1B) inhibitors, or in Japanese patent JP 2002308870, in which compounds are described, which act as Staphylococcus aureus inhibitors. Further derivatives are described in Armyanskii Khimicheskii Zhurnal 1987, 40(9), 581-7. These references do not disclose any PkNG inhibitory activity for these compounds.
In WO 01/98290 thiophene derivatives are described as active kinase inhibitors.
One important feature for pharmaceutical active agents in general is that these agents have a high degree of metabolitic stability. It was found that the compounds described in PCT/EP03/03697, while being pharmaceutically active as PkNG inhibitors, left room for further increase of metabolitic stability.
The present invention also provides a method for preventing or treating infections and diseases, especially virally or bacterially induced diseases or infections, more specially infections induced by bacteria of the genus legionella such as legionaires disease, mycobacteria-induced infections (including opportunistic infections) in mammals (including humans), which method comprises administering to the mammal an pharmaceutically effective amount of the compounds of the present invention to treat a infection or disease. Especially, the method is used for the treatment of tuberculosis, but also for other mycobacteria-induced infections like leprosy or mycobacteria-induced meningitis.
Further aspects of the present invention relate to the use of the compounds of general formula (I) for the preparation of a pharmaceutical composition useful for prophylaxis and/or treatment of infectious diseases including opportunistic diseases, prion diseases, immunological diseases, autoimmune diseases, bipolar and clinical disorders, cardiovascular diseases, cell proliferative diseases, diabetes, inflammation, transplant rejections, erectile dysfunction, neurodegenerative diseases, and stroke.
Infectious Diseases Including Opportunistic Infections
In yet another aspect of the present invention, the compounds according to the general formula (I) are for the preparation of a pharmaceutical composition for the prophylaxis and/or treatment of infectious diseases, including opportunistic diseases and opportunistic infections. The term infectious diseases comprises infections caused by viruses, bacteria, prions, fungi, and/or parasites.
Especially, virally induced infectious diseases, including opportunistic diseases are addressed. In a preferred embodiment of this aspect, the virally induced infectious diseases, including opportunistic diseases, are caused by retroviruses, human endogenous retroviruses (HERVs), hepadnaviruses, herpesviruses, flaviviridae, and/or adenoviruses. Preferably, the retroviruses are selected from lentiviruses or oncoretroviruses, wherein the lentivirus is preferably selected from the group comprising: HIV-1, HIV-2, feline immunodeficiency virus (FIV), bovine immunodeficiency virus (BIV), sivian immunodeficiency viruses (SIVs), chimeras of HIV and SIV (SHIV), caprine arthritis encephalitis virus (CAEV), visna/maedi virus (VMV) or equine infectious anemia virus (EIAV), preferably HIV-1 and HIV-2, and the oncoretrovirus is preferably selected from HTLV-I, HTLV-II or bovine leukemia virus (BLV), preferably HTLV-I and HTLV-II.
The hepadnavirus is preferably selected from HBV, ground squirrel hepatitis virus (GSHV) or woodchuck hepatitis virus (WHV), preferably HBV, the herpesvirus is selected from the group comprising: Herpes simplex virus I (HSV I), herpes simplex virus II (HSV II), Epstein-Barr virus (EBV), varicella zoster virus (VZV), human cytomegalovirus (HCMV) or human herpesvirus 8 (HHV-8), preferably HCMV, and the flaviviridae is selected from HCV, West nile or Yellow Fever.
It is to be understood, that all the viruses mentioned above, also comprise drug resistant virus strains.
Examples of infective diseases are AIDS, Alveolar Hydatid Disease (AHD, Echinococcosis), Amebiasis (Entamoeba histolytica Infection), Angiostrongylus Infection, Anisakiasis, Anthrax, Babesiosis (Babesia Infection), Balantidium Infection (Balantidiasis), Baylisascaris Infection (Raccoon Roundworm), Bilharzia (Schistosomiasis), Blastocystis hominis Infection (Blastomycosis), Boreliosis, Botulism, Brainerd Diarrhea, Brucellosis, BSE (Bovine Spongiform Encephalopathy), Candidiasis, Capillariasis (Capillaria Infection), CFS (Chronic Fatigue Syndrome), Chagas Disease (American Trypanosomiasis), Chickenpox (Varicella-Zoster virus), Chlamydia pneumoniae Infection, Cholera, Chronic Fatigue Syndrome, CJD (Creutzfeldt-Jakob Disease), Clonorchiasis (Clonorchis Infection), CLM (Cutaneous Larva Migrans, Hookworm Infection), Coccidioidomycosis, Conjunctivitis, Coxsackievirus A16 (Hand, Foot and Mouth Disease), Cryptococcosis, Cryptosporidium Infection (Cryptosporidiosis), Culex mosquito (Vector of West Nile Virus), Cutaneous Larva Migrans (CLM), Cyclosporiasis (Cyclospora Infection), Cysticercosis (Neurocysticercosis), Cytomegalovirus Infection, Dengue/Dengue Fever, Dipylidium Infection (Dog and Cat Flea Tapeworm), Ebola Virus Hemorrhagic Fever, Echinococcosis (Alveolar Hydatid Disease), Encephalitis, Entomoeba coli Infection, Entomoeba dispar Infection, Entomoeba hartmanni Infection, Entomoeba histolytica Infection (Amebiasis), Entomoeba polecki Infection, Enterobiasis (Pinworm Infection), Enterovirus Infection (Non-Polio), Epstein-Barr Virus Infection, Escherichia coli Infection, Foodborne Infection, Foot and mouth Disease, Fungal Dermatitis, Gastroenteritis, Group A streptococcal Disease, Group B streptococcal Disease, Hansen's Disease (Leprosy), Hantavirus Pulmonary Syndrome, Head Lice Infestation (Pediculosis), Helicobacter pylori Infection, Hematologic Disease, Hendra Virus Infection, Hepatitis (HCV, HBV), Herpes Zoster (Shingles), HIV Infection, Human Ehrlichiosis, Human Parainfluenza Virus Infection, Influenza, Isosporiasis (Isospora Infection), Lassa Fever, Leishmaniasis, Kala-azar (Kala-azar, Leishmania Infection), Leprosy, Lice (Body lice, Head lice, Pubic lice), Lyme Disease, Malaria, Marburg Hemorrhagic Fever, Measles, Meningitis, Mosquito-borne Diseases, Mycobacterium avium Complex (MAC) Infection, Naegleria Infection, Nosocomial Infections, Nonpathogenic Intestinal Amebae Infection, Onchocerciasis (River Blindness), Opisthorciasis (Opisthorcis Infection), Parvovirus Infection, Plague, PCP (Pneumocystis carinii Pneumonia), Polio, Q Fever, Rabies, Respiratory Syncytial Virus (RSV) Infection, Rheumatic Fever, Rift Valley Fever, River Blindness (Onchocerciasis), Rotavirus Infection, Roundworms Infection, Salmonellosis, Salmonella Enteritidis, Scabies, Shigellosis, Shingles, Sleeping Sickness, Smallpox, Streptococcal Infection, Tapeworm Infection (Taenia Infection), Tetanus, Toxic Shock Syndrome, Tuberculosis, Ulcers (Peptic Ulcer Disease), Valley Fever, Vibrio parahaemolyticus Infection, Vibrio vulnificus Infection, Viral Hemorrhagic Fever, Warts, Waterborne infectious Diseases, West Nile Virus Infection (West Nile Encephalitis), Whooping Cough, Yellow Fever.
Bacterial Infections
As described above, the compounds according to the general formula (I) are also useful for the preparation of a pharmaceutical composition for prophylaxis and/or treatment of bacterially induced infectious diseases, including opportunistic diseases and opportunistic infections, wherein the bacterially induced infectious diseases, including opportunistic diseases, are selected from tuberculosis, leprosy or mycobacteria-induced meningitis. One advantage of the inventive compounds disclosed herein is there use against drug resistant bacterial strains.
Prion Diseases
Another aspect of the present invention is directed to the use of at least one compound of the general formula (I) and/or pharmaceutically acceptable salts thereof for prophylaxis and/or treatment of prion diseases.
Prions are infectious agents, which do not have a nucleic acid genome. It seems that a protein alone is the infectious agent. A prion has been defined as “small proteinaceous infectious particle, which resists inactivation, by procedures that modify nucleic acids”. The discovery that proteins alone can transmit an infectious disease has come as a considerable surprise to the scientific community. Prion diseases are often called “transmissible spongiform encephalopathies”, because of o the post mortem appearance of the brain with large vacuoles in the cortex and cerebellum. Probably most mammalian species develop these diseases. Prion diseases are a group of neurodegenerative disorders of humans and animals and the prion diseases can manifest as sporadic, genetic or infectious disorders. Examples for prion diseases acquired by exogenous infection are the Bovine spongiform encephalitis (BSE) of cattle and the new variant of Creutzfeld-Jakob disease (vCJD) caused by BSE as well as scrapie of animals. Examples of human prion diseases include kuru, sporadic Creutzfeldt-Jakob disease (sCJD), familial CJD (fCJD), iatrogenic CJD (iCJD), Gerstmann-Sträussler-Scheinker (GSS) disease, fatal familial insomnia (FFI), and especially the new variant CJD (nvCJD or vCJD).
The name “prion” is used to describe the causative agents, which underlie the transmissible spongiform encephalopathies. A prion is proposed to be a novel infectious particle that differs from viruses and viroids. It is composed solely of one unique protein that resists most inactivation procedures such as heat, radiation, and proteases. The latter characteristic has led to the term protease-resistant isoform of the prion protein. The protease-resistant isoform has been proposed to slowly catalyze the conversion of the normal prion protein into the abnormal form.
The term “isoform” in the context of prions means two proteins with exactly the same amino acid sequence, that are folded into molecules with dramatically different tertiary structures. The normal cellular isoform of the prion protein (PrPC) has a high a-helix content, a low b-sheet content, and is sensitive to protease digestion. The abnormal, disease-causing isoform (PrPSc)has a lower a-helix content, a much higher b-sheet content, and is much more resistant to protease digestion.
As used herein the term “prion diseases” refers to transmissible spongiform encephalopathies. Examples for prion diseases comprise Scrapie (sheep, goat), TME (transmissible mink encephalopathy; mink), CWD (chronic wasting disease; muledeer, deer, elk), BSE (bovine spongiform encephalopathy; cows, cattles), CJD (Creutzfeld-Jacob Disease), vCJD, GSS (Gerstmann-Sträussler-Scheinker syndrome), FFI (Fatal familial Insomnia), Kuru, and Alpers Syndrome. Preferred are BSE, vCJD, and CJD.
Immunological Diseases
Another aspect of the present invention is directed to the use of at least one compound of the general formula (I) and/or pharmaceutically acceptable salts thereof for prophylaxis and/or treatment of immunological diseases, neuroimmunological diseases, and autoimmune diseases.
Immunological diseases are, for instance, asthma and diabetes, rheumatic and autoimmune diseases, AIDS, rejection of transplanted organs and tissues (cf. below), rhinitis, chronic obstructive pulmonary diseases, osteoporisis, ulcerative colitis, sinusitis, lupus erythematosus, recurrent infections, atopic dermatitis/eczema and occupational allergies, food allergies, drug allergies, severe anaphylactic reactions, anaphylaxis, and other manifestations of allergic disease, as well as uncommon problems such as primary immunodeficiencies, including antibody deficiency states, cell mediated immunodeficiencies (e.g., severe combined immunodeficiency, DiGeorge syndrome, Hyper-IgE syndrome, Wiskott-Aldrich syndrome, ataxia-telangiectasia), immune mediated cancers, and white cell defects.
In autoimmune diseases, such as systemic lupus erythematosus, rheumatoid arthritis (RA), multiple sclerosis (MS), immune-mediated or type 1 diabetes mellitus, immune mediated glomerulonephritis, scleroderma, pernicious anemia, alopecia, pemphigus, pemphigus vulgaris, myasthenia gravis, inflammatory bowel diseases, Crohn's disease, psoriasis, autoimmune thyroid diseases, and Hashimoto's disease, dermatomyositis, goodpastture syndrome, myasthenia gravis pseudoparalytica, ophtalmia sympatica, phakogene uveitis, chronical agressivce hepatitis, primary billiary cirrhosis, autoimunehemolytic anemy, Werlof disease, specific cells uncontrollably attack the body's own tissues and organs (autoimmunity), producing inflammatory reactions and other serious symptoms and diseases.
Hashimoto's thyroiditis is one of the most common autoimmune diseases. “Autoimmune disease” refers to a category of more than 80 chronic illnesses, each very different in nature, that can affect everything from the endocrine glands (like the thyroid) to organs like the kidneys, as well as to the digestive system.
There are many different autoimmune diseases, and they can each affect the body in different ways. For example, the autoimmune reaction is directed against the brain in multiple sclerosis and the gut in Crohn's disease. In other autoimmune diseases such as systemic lupus erythematosus (lupus), affected tissues and organs may vary among individuals with the same disease. One person with lupus may have affected skin and joints whereas another may have affected skin, kidney, and lungs. Ultimately, damage to certain tissues by the immune system may be permanent, as with destruction of insulin-producing cells of the pancreas in Type 1 diabetes mellitus.
Bipolar and Clinical Disorders
Another aspect of the present invention is directed to the use of at least one compound of the general formula (I) and/or pharmaceutically acceptable salts thereof for prophylaxis and/or treatment of bipolar and clinical disorders.
The term “bipolar and clinical disorders” shall refer to adjustment disorders, anxiety disorders, delirium, dementia, amnestic and other cognitive disorders, disorders usually first diagnosed in infancy, childhood, or adolescence, dissociative disorders, eating disorders, factitious disorders, impulse-control disorders, mental disorders due to a general medical condition, mood disorders, other conditions that may be a focus of clinical attention, personality disorders, schizophrenia and other psychotic disorders, sexual and gender identity disorders, sleep disorders, somatoform disorders, substance-related disorders, generalized anxiety disorder, panic disorder, phobia, agoraphobia, obsessive-compulsive disorder, stress, acute stress disorder, anxiety neurosis, nervousness, phobia, posttraumatic stress disorder, posttraumatic stress disorder (PTSD), abuse, obsessive-compulsive disorder (OCD), manic depressive psychosis, specific phobias, social phobia, adjustment disorder with anxious features.
Examples for anxiety disorders are: acute stress disorder, agoraphobia without history of panic disorder, anxiety disorder due to general medical condition, generalized anxiety disorder, obsessive-compulsive disorder, panic disorder with agoraphobia, panic disorder without agoraphobia, posttraumatic stress disorder, specific phobia, social phobia, substance-induced anxiety disorder.
Examples for delirium, dementia, amnestic and other cognitive disorders are: delirium due to a general medical condition, substance intoxication delirium, substance withdrawal delirium, delirium due to multiple etiologies, Alzheimer's, Creutzfeldt-Jakob disease, head trauma, Huntington's disease, HIV disease, Parkinson's disease, Pick's disease, substance-induced persisting, vascular, dementia due to other general medical conditions, dementia due to multiple etiologies, amnestic disorder due to a general medical condition, substance-induced persisting amnestic disorder.
Examples for disorders usually first diagnosed in infancy, childhood, or adolescence are: mental retardation, learning disorders, mathematics disorder, reading disorder, disorder of written expression, learning disorder, motor skills disorders, developmental coordination disorder, communication disorders, expressive language disorder, phonological disorder, mixed receptive-expressive language disorder, stuttering, pervasive developmental disorders, Asperger's disorder, autistic disorder, childhood disintegrative disorder, Rett's disorder, pervasive developmental disorder, attention-deficit/hyperactivity disorder (ADHD), conduct disorder, oppositional defiant disorder, feeding disorder of infancy or early childhood, pica, rumination disorder, tic disorders, chronic motor or vocal tic disorder, Tourette's syndrome, elimination disorders, encopresis, enuresis, selective mutism, separation anxiety disorder, reactive attachment disorder of infancy or early childhood, stereotypic movement disorder.
Examples for dissociative disorders are: dissociative amnesia, depersonalization disorder, dissociative fugue and dissociative identity disorder.
Examples for eating disorders are anorexia nervosa and bulimia nervosa.
Examples for mood disorders are: mood episodes, major depressive episode, hypomanic episode, manic episode, mixed episode, depressive disorders, dysthymic disorder, major depressive disorder, single episode, recurrent, bipolar disorders, bipolar I disorder, bipolar II disorder, cyclothymic disorder, mood disorder due to a general medical condition, substance-induced mood disorder.
Examples for schizophrenia and other psychotic disorders are: schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, shared psychotic disorder, psychotic disorder due to a general medical condition, delusions, hallucinations, substance-induced psychotic disorder.
Examples for sexual and gender identity disorders are: female sexual arousal disorder, orgasmic disorders, premature ejaculation, sexual pain disorders, dyspareunia, vaginismus, sexual dysfunction due to a general medical condition, female dyspareunia, female hypoactive sexual desire disorder, male erectile disorder, male hypoactive sexual desire disorder, male dyspareunia, other female sexual dysfunction, other male sexual dysfunction, substance-induced sexual dysfunction, sexual dysfunction, exhibitionism, fetishism, frotteurism, pedophilia, masochism, sadism, transvestic fetishism, voyeurism, paraphilia, gender identity disorder.
Examples for sleep disorders are: dyssomnias, breathing-related sleep disorder, circadian rhythm sleep disorder, hypersomnia, hypersomnia related to another mental disorder, insomnia, insomnia related to another mental disorder, narcolepsy, dyssomnia, parasomnias, nightmare disorder, sleep terror disorder, sleepwalking disorder, parasomnia.
Examples for somatoform disorders are: body dysmorphic disorder, conversion disorder, hypochondriasis, pain disorder, somatization disorder, undifferentiated somatoform disorder.
Examples for substance-related disorders are: alcohol related disorders, amphetamine related disorders, caffeine related disorders, cannabis related disorders, cocaine related disorders, hallucinogen related disorders, inhalant related disorders, nicotine related disorders, opioid related disorders, psychotic disorder, psychotic disorder, phencyclidine-related disorder, abuse, persisting amnestic disorder, anxiety disorder, persisting dementia, dependence, intoxication, intoxication delirium, mood disorder, psychotic/disorder, withdrawal, withdrawal delirium, sexual dysfunction, sleep disorder.
Cardiovascular Diseases
The inventive compounds are also useful for prophylaxis and/or treatment of cardiovascular diseases such as adult congenital heart disease, aneurysm, stable angina, unstable angina, angina pectoris, angioneurotic edema, aortic valve stenosis, aortic aneurysm, arrhythmia, arrhythmogenic right ventricular dysplasia, arteriosclerosis, arteriovenous malformations, atrial fibrillation, Behcet syndrome, bradycardia, cardiac tamponade, cardiomegaly, congestive cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, cardiovascular disease prevention, carotid stenosis, cerebral hemorrhage, Churg-Strauss syndrome, diabetes, Ebstein's Anomaly, Eisenmenger complex, cholesterol embolism, bacterial endocarditis, fibromuscular dysplasia, congenital heart defects, heart diseases, congestive heart failure, heart valve diseases, heart attack, epidural hematoma, hematoma, subdural, Hippel-Lindau disease, hyperemia, hypertension, pulmonary hypertension, hypertrophic growth, left ventricular hypertrophy, right ventricular hypertrophy, hypoplastic left heart syndrome, hypotension, intermittent claudication, ischemic heart disease, Klippel-Trenaunay-Weber syndrome, lateral medullary syndrome, long QT syndrome mitral valve prolapse, moyamoya disease, mucocutaneous lymph node syndrome, myocardial infarction, myocardial ischemia, myocarditis, pericarditis, peripheral vascular diseases, phlebitis, polyarteritis nodosa, pulmonary atresia, Raynaud disease, restenosis, Sneddon syndrome, stenosis, superior vena cava syndrome, syndrome X, tachycardia, Takayasu's arteritis, hereditary hemorrhagic telangiectasia, telangiectasis, temporal arteritis, tetralogy of fallot, thromboangiitis obliterans, thrombosis, thromboembolism, tricuspid atresia, varicose veins, vascular diseases, vasculitis, vasospasm, ventricular fibrillation, Williams syndrome, peripheral vascular disease, varicose veins and leg ulcers, deep vein thrombosis, Wolff-Parkinson-White syndrome.
Preferred are adult congenital heart disease, aneurysms, angina, angina pectoris, arrhythmias, cardiovascular disease prevention, cardiomyopathies, congestive heart failure, myocardial infarction, pulmonary hypertension, hypertrophic growth, restenosis, stenosis, thrombosis and arteriosclerosis.
Proliferative Disease
In yet another preferred embodiment, the cell proliferative disease is cancer, which is preferably selected from the group comprising:
The proliferation disorders and cancers are preferably selected from the group comprising adenocarcinoma, choroidal melanoma, acute leukemia, acoustic neurinoma, ampullary carcinoma, anal carcinoma, astrocytoma, basal cell carcinoma, pancreatic cancer, desmoid tumor, bladder cancer, bronchial carcinoma, breast cancer, Burkitt's lymphoma, corpus cancer, CUP-syndrome (carcinoma of unknown primary), colorectal cancer, small intestine cancer, small intestinal tumors, ovarian cancer, endometrial carcinoma, ependymoma, epithelial cancer types, Ewing's tumors, gastrointestinal tumors, gastric cancer, gallbladder cancer, gall bladder carcinomas, uterine cancer, cervical cancer, cervix, glioblastomas, gynecologic tumors, ear, nose and throat tumors, hematologic neoplasias, hairy cell leukemia, urethral cancer, skin cancer, skin testis cancer, brain tumors (gliomas), brain metastases, testicle cancer, hypophysis tumor, carcinoids, Kaposi's sarcoma, laryngeal cancer, germ cell tumor, bone cancer, colorectal carcinoma, head and neck tumors (tumors of the ear, nose and throat area), colon carcinoma, craniopharyngiomas, oral cancer (cancer in the mouth area and on lips), cancer of the central nervous system, liver cancer, liver metastases, leukemia, eyelid tumor, lung cancer, lymph node cancer (Hodgkin's/Non-Hodgkin's), lymphomas, stomach cancer, malignant melanoma, malignant neoplasia, malignant tumors gastrointestinal tract, breast carcinoma, rectal cancer, medulloblastomas, melanoma, meningiomas, Hodgkin's disease, mycosis fungoides, nasal cancer, neurinoma, neuroblastoma, kidney cancer, renal cell carcinomas, non-Hodgkin's lymphomas, oligodendroglioma, esophageal carcinoma, osteolytic carcinomas and osteoplastic carcinomas, osteosarcomas, ovarial carcinoma, pancreatic carcinoma, penile cancer, plasmocytoma, prostate cancer, pharyngeal cancer, rectal carcinoma, retinoblastoma, vaginal cancer, thyroid carcinoma, Schneeberger disease, esophageal cancer, spinalioms, T-cell lymphoma (mycosis fungoides), thymoma, tube carcinoma, eye tumors, urethral cancer, urologic tumors, urothelial carcinoma, vulva cancer, wart appearance, soft tissue tumors, soft tissue sarcoma, Wilm's tumor, cervical carcinoma and tongue cancer.
Preferred are the following cancer types: bladder, breast, central nervous system, colon, gastric, lung, kidney, melanoma, head and neck, ovarian, cervix, glioblastoma, pancreas, prostate, stomach, skin testis, leukemia, Hodgkin's lymphoma, liver and renal cancer.
Diabetes
In yet another preferred embodiment, said diabetes is selected from Type I diabetes or Type II diabetes.
Inflammation
In yet another preferred embodiment, said inflammation is mediated preferably by the cytokines TNF-α, IL-1β, GM-CSF, IL-6 and/or IL-8.
As described above, the compounds according to general formula (I) are pharmaceutically active agents for prophylaxis and/or treatment of inflammatory diseases. Thus, these compounds are used for the manufacture of a pharmaceutical formulation for prophylaxis and/or treatment of inflammations and inflammatory diseases in mammals, including humans.
Inflammatory diseases can emanate from infectious and non-infectious inflammatory conditions which may result from infection by an invading organism or from irritative, traumatic, metabolic, allergic, autoimmune, or idiopathic causes as shown in the following list.
Thus, the compounds disclosed herein can be used for prophylaxis and/or treatment of inflammations caused by invading organisms such as viruses, bacteria, prions, and parasites as well as for prophylaxis and/or treatment of inflammations caused by irritative, traumatic, metabolic, allergic, autoimmune, or idiopathic reasons.
Consequently, the disclosed compounds are useful for prophylaxis and/or treatment of inflammatory diseases which are initiated or caused by viruses, parasites, and bacteria which are connected to or involved in inflammations.
The following bacteria are known to cause inflammatory diseases: mycoplasma pulmonis (causes e.g. chronic lung diseases (CLD), murine chronic respiratory disease), ureaplasma urealyticum (causes pneumonia in newborns), mycoplasma pneumoniae and chlamydia pneumoniae (cause chronic asthma), C. pneumoniae (causes atherosclerosis, pharyngitis to pneumonia with empyema, human coronary heart disease), Helicobacter pylori (human coronary heart disease, stomach ulcers).
The following viruses are known to cause inflammatory diseases: herpesviruses especially cytomegalovirus (causes human coronary heart disease).
The compounds disclosed herein are useful for prophylaxis and/or treatment of inflammatory diseases caused and/or induced and/or initiated and/or enhanced by the afore-mentioned bacteria or viruses.
Furthermore, the compounds of formula (I) are useful for prophylaxis and/or treatment of inflammatory diseases of the central nervous system (CNS), inflammatory rheumatic diseases, inflammatory diseases of blood vessels, inflammatory diseases of the middle ear, inflammatory bowel diseases, inflammatory diseases of the skin, inflammatory disease uveitis, inflammatory diseases of the larynx.
Examples for inflammatory diseases of the central nervous system (CNS) are algal disorders, protothecosis, bacterial disorders, abscessation, bacterial meningitis, idiopathic inflammatory disorders, eosinophilic meningoencephalitis, feline polioencephalomyelitis, granulomatous meningoencephalomyelitis, meningitis, steroid responsive meningitis-arteritis, miscellaneous meningitis/meningoencephalitis, meningoencephalitis in greyhounds, necrotizing encephalitis, pug dog encephalitis, pyogranulomatous meningoencephalomyelitis, shaker dog disease, mycotic diseases of the CNS, parasitic encephalomyelitis, prion protein induced diseases, feline spongiform encephalopathy, protozoal encephalitis-encephalomyelitis, toxoplasmosis, neosporosis, sarcocystosis, encephalitozoonosis, trypanosomiasis, acanthamebiasis, babesiosis, leishmaniasis, rickettsial disorders, rocky mountain spotted fever, canine ehrlichiosis, salmon poisoning, viral disorders, aujeszky's disease, borna disease, canine herpes virus encephalomyelitis, canine distemper encephalomyelitis, canine distemper encephalomyelitis in immature animals, multifocal distemper encephalomyelitis in mature animals, old dog encephalitis, chronic relapsing encephalomyelitis, post-vaccinal canine distemper encephalitis, feline immunodeficiency virus, feline infectious peritonitis, feline leukemia virus, infectious canine hepatitis, La Crosse virus encephalitis, parvovirus encephalitis, rabies, post-vaccinal rabies, tick-borne encephalitis in dogs.
Examples for inflammatory rheumatic diseases are rheumatoid arthritis, scleroderma, lupus, polymyositis, dermatomyositis, psoriatic arthritis, ankylosing spondylitis, Reiters's syndrome, juvenile rheumatoid arthritis, bursitis, tendinitis (tendonitis), and fibromyositis.
Examples for inflammatory diseases of blood vessels are vasculitis, autoantibodies in vasculitis, microscopic polyangiitis, giant cell arteritis, Takayasu's arteritis, vasculitis of the central nervous system, thromboangiitis obliterans (Buerger's Disease), vasculitis secondary to bacterial, fungal, and parasitic infection, vasculitis and rheumatoid arthritis, vasculitis in systemic lupus erythematosus, vasculitis in the idiopathic inflammatory myopathies, relapsing polychondritis, systemic vasculitis in sarcoidosis, vasculitis and malignancy, and drug-induced vasculitis.
Examples for inflammatory diseases of the middle ear are acute suppurative otitis media, bullous myringitis, granular myringitis, and chronic suppurative otitis media, which can manifest as mucosal disease, cholesteatoma, or both.
Examples for inflammatory bowel diseases are ulcerative colitis, Crohn's disease.
Examples for inflammatory diseases of the skin are acute inflammatory dermatoses, urticaria (hives), spongiotic dermatitis, allergic contact dermatitis, irritant contact dermatitis, atopic dermatitis, erythemal multiforme (EM minor), Stevens-Johnson syndrome (SJS, EM major), toxic epidermal necrolysis (TEN), chronic inflammatory dermatoses, psoriasis, lichen planus, discoid lupus erythematosus, and acne vulgaris
Uveitis are inflammations located in and/or on the eye and may be associated with inflammation elsewhere in the body. In most circumstances, patients who have uveitis as part of a disease elsewhere in the body are aware of that illness. The majority of patients with uveitis do not have an apparent associated systemic illness. Causes of uveitis can be infectious causes, masquerade syndromes, suspected immune-mediated diseases, and/or syndromes confined primarily to the eye.
The following viruses are associated with inflammations: human immunodeficiency virus-I, herpes simplex virus, herpes zoster virus, and cytomegalovirus.
Bacterial or spirochetal caused, induced, initiated and/or enhanced inflammations are tuberculosis, leprosy, proprionobacterium, syphilis, Whipple's disease, leptospirosis, brucellosis, and lyme disease.
Parasitic (protozoan or helminthic) caused, induced, initiated and/or enhanced inflammations are toxoplasmosis, acanthameba, toxocariasis, cysticercosis, onchocerciasis.
Examples of inflammatory diseases caused, induced, initiated and/or enhanced by fungi are histoplasmosis, coccidioidomycosis, candidiasis, aspergillosis, sporotrichosis, blastomycosis, and cryptococcosis.
Masquerade syndromes are, for instance, leukemia, lymphoma, retinitis pigmentosa, and retinoblastoma.
Suspected immune-mediated diseases can be selected from the group comprising ankylosing spondylitis, Behcet's disease, Crohn's disease, drug or hypersensitivity reaction, interstitial nephritis, juvenile rheumatoid arthritis, Kawasaki's disease, multiple sclerosis, psoriatic arthritis, Reiter's syndrome, relapsing polychondritis, sarcoidosis, Sjogren's syndrome, systemic lupus erythematosus, ulcerative colitis, vasculitis, vitiligo, Vogt Koyanagi Harada syndrome.
Syndromes confined primarily to the eye are, for instance, acute multifocal placoid pigmentary epitheliopathy, acute retinal necrosis, birdshot choroidopathy, Fuch's heterochromic cyclitis, glaucomatocyclitic crisis, lens-induced uveitis, multifocal choroiditis, pars planitis, serpiginous choroiditis, sympathetic ophthalmia, and trauma.
Examples for inflammatory diseases of the larynx are gastroesophageal (laryngopharyngeal) reflux disease, pediatric laryngitis, acute laryngeal infections of adults, chronic (granulomatous) diseases, allergic, immune, and idiopathic disorders and miscellaneous inflammatory conditions.
Pediatric laryngitis is known as acute (viral or bacterial) infection such as laryngotracheitis (croup), supraglottitis (epiglottitis), diphtheria, and noninfectious causes are for example spasmodic croup and traumatic laryngitis.
Acute laryngeal infections of adults are, for instance, viral laryngitis, common upper respiratory infection, laryngotracheitis, herpes simplex, bacterial laryngitis, supraglottitis, laryngeal abscess, and gonorrhea.
Chronic (granulomatous) diseases can be selected from the group comprising bacterial diseases, tuberculosis, leprosy, scleroma, actinomycosis, tularemia, glanders, spirochetal (syphilis) diseases, mycotic (fungal) diseases, candidiasis, blastomycosis, histoplasmosis, coccidiomycosis, aspergillosis, idiopathic diseases, sarcoidosis, and Wegener's granulomatosis.
Allergic, immune, and idiopathic disorders are, for example, hypersensitivity reactions, angioedema, Stevens-Johnson syndrome, immune and idiopathic disorders, infections of the immunocompromised host, rheuatoid arthritis, systeic lupus erythematosus, cicatricial pemphigoid, relapsing polychondritis, Sjogren's syndrome, and amyloidosis.
Miscellaneous inflammatory conditions are, for instance, parasitic infections, trichinosis, leishmaniasis, schistosomiasis, syngamus laryngeus, inhalation laryngitis, acute (thermal) injury, pollution and inhalant allergy, carcinogens, radiation injury, radiation laryngitis, radionecrosis, vocal abuse, vocal-cord hemorrhage, muscle tension dysphonias, and contact ulcer and granuloma.
Transplant Rejection
Transplant rejection is when a transplant recipient's immune system attacks a transplanted organ or tissue. No two people (except identical twins) have identical tissue antigens. Therefore, in the absence of immunosuppressive drugs, organ and tissue transplantation would almost always cause an immune response against the foreign tissue (rejection), which would result in destruction of the transplant. Though tissue typing ensures that the organ or tissue is as similar as possible to the tissues of the recipient, unless the donor is an identical twin, no match is perfect and the possibility of organ/tissue rejection remains.
The inventive compounds of general formula (I) are used as immunosuppressive drugs and/or anti-rejection drugs in order to prevent transplant rejection.
One example of transplant rejection is the graft-versus-host-disease (GVHD) that can occur following bone marrow transplant. The donor's immune cells in the transplanted marrow make antibodies against the host's (transplant patient's) tissues and attack the patient's vital organs. Transplant rejections (also known as graft rejection or tissue/organ rejection) may commonly occur when tissue or organs, which need blood supply, are transplanted. Said organs comprise especially inner organs such as heart, heart-lungs, lungs, liver, kidney, pancreas, spleen, skin, tissue, bone marrow, spinal marrow, hormone producing glands, gonads and gonadal glands.
Neurodegenerative Diseases
Another aspect of the present invention is directed to the use of at least one compound of the general formula (I) and/or pharmaceutically acceptable salts thereof for prophylaxis and/or treatment of neurodegeneration and neurodegenerative disorders.
Among the hundreds of different neurodegenerative disorders, the attention has been given only to a handful, including Alzheimer disease, Parkinson disease, Huntington disease, and amyotrophic lateral sclerosis.
It is worth to mention that the same neurodegenerative process can affect different areas of the brain, making a given disease appear very different from a symptomatic standpoint.
Neurodegenerative disorders of the central nervous system (CNS) can be grouped into diseases of the cerebral cortex (Alzheimer disease), the basal ganglia (Parkinson disease), the brain-stem and cerebellum, or the spinal cord (amyotrophic lateral sclerosis).
Examples for neurodegeneration and neurodegenerative disorders are Alzheimer disease, Parkinson disease, Huntington disease, amyotrophic lateral sclerosis, AIDS-related dementia, retinitis pigmentosa, spinal muscular atrophy and cerebrellar degeneration, fragile X-associated tremor/ataxia syndrome (FXTAS), progressive supranuclear palsy (PSP), and striatonigral degeneration (SND), which is included with olivopontocerebellear degeneration (OPCD), and Shy Drager syndrome (SDS) in a syndrome known as multiple system atrophy (MSA).
According to a still further aspect, the present invention refers to pharmaceutical compositions comprising at least one compound according to the present invention as an active ingredient together with at least one pharmaceutically acceptable (i.e. non-toxic) carrier, excipient and/or diluent. The pharmaceutical compositions of the present invention can be prepared in a conventional solid or liquid carrier or diluent and a conventional pharmaceutically-made adjuvant at suitable dosage level in a known way. The preferred preparations are adapted for oral application. These administration forms include, for example, pills, tablets, film tablets, coated tablets, capsules, powders and deposits.
Furthermore, the present invention also includes pharmaceutical preparations for parenteral application, including dermal, intradermal, intragastral, intracutan, intravasal, intravenous, intramuscular, intraperitoneal, intranasal, intravaginal, intrabuccal, percutan, rectal, subcutaneous, sublingual, topical, or transdermal application, which preparations in addition to typical vehicles and/or diluents contain at least one compound according to the present invention and/or a pharmaceutical acceptable salt thereof as active ingredient.
The pharmaceutical compositions according to the present invention containing at least one compound according to the present invention, e.g. one 4,7-dihydro-5H-thieno[2,3-c]thiopyran-3-carboxylic acid amide and/or one 4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide, and/or one 4,7-dihydro-5H-thieno[2,3-c]pyran derivative or analogues thereof as set out in general formula (I) in independent claim 1 or claims dependent thereon, and/or a pharmaceutical acceptable salt thereof as active ingredient will typically be administered together with suitable carrier materials selected with respect to the intended form of administration, i.e. for oral administration in the form of tablets, capsules (either solid filled, semi-solid filled or liquid filled), powders for constitution, gels, elixirs, dispersable granules, syrups, suspensions, and the like, and consistent with conventional pharmaceutical practices. For example, for oral administration in the form of tablets or capsules, the active drug component may be combined with any oral non-toxic pharmaceutically acceptable carrier, preferably with an inert carrier like lactose, starch, sucrose, cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, talc, mannitol, ethyl alcohol (liquid filled capsules) and the like. Moreover, suitable binders, lubricants, disintegrating agents and coloring agents may also be incorporated into the tablet or capsule. Powders and tablets may contain about 5 to about 95 weight % of the heterobicyclic compound and/or the respective pharmaceutically active salt as active ingredient.
Suitable binders include starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethylcellulose, polyethylene glycol and waxes. Among suitable lubricants there may be mentioned boric acid, sodium benzoate, sodium acetate, sodium chloride, and the like. Suitable disintegrants include starch, methylcellulose, guar gum, and the like. Sweetening and flavoring agents as well as preservatives may also be included, where appropriate. The disintegrants, diluents, lubricants, binders etc. are discussed in more detail below.
Moreover, the pharmaceutical compositions of the present invention may be formulated in sustained release form to provide the rate controlled release of any one or more of the components or active ingredients to optimise the therapeutic effect(s), e.g. antihistaminic activity and the like. Suitable dosage forms for sustained release include tablets having layers of varying disintegration rates or controlled release polymeric matrices impregnated with the active components and shaped in tablet form or capsules containing such impregnated or encapsulated porous polymeric matrices.
Liquid form preparations include solutions, suspensions, and emulsions. As an example, there may be mentioned water or water/propylene glycol solutions for parenteral injections or addition of sweeteners and opacifiers for oral solutions, suspensions, and emulsions. Liquid form preparations may also include solutions for intranasal administration.
Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be present in combination with a pharmaceutically acceptable carrier such as an inert, compressed gas, e.g. nitrogen.
For preparing suppositories, a low melting wax, such as a mixture of fatty acid glycerides like cocoa butter is melted first, and the active ingredient is then dispersed homogeneously therein e.g. by stirring. The molten, homogeneous mixture is then poured into conveniently sized moulds, allowed to cool, and thereby solidified.
Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions, and emulsions.
The compounds according to the present invention may also be delivered transdermally. The transdermal compositions may have the form of a cream, a lotion, an aerosol and/or an emulsion and may be included in a transdermal patch of the matrix or reservoir type as is known in the art for this purpose.
The term capsule as recited herein refers to a specific container or enclosure made e.g. of methyl cellulose, polyvinyl alcohols, or denatured gelatins or starch for holding or containing compositions comprising the active ingredient(s). Capsules with hard shells are typically made of blended of relatively high gel strength gelatins from bones or pork skin. The capsule itself may contain small amounts of dyes, opaquing agents, plasticisers and/or preservatives.
Under tablet a compressed or moulded solid dosage form is understood which comprises the active ingredients with suitable diluents. The tablet may be prepared by compression of mixtures or granulations obtained by wet granulation, dry granulation, or by compaction well known to a person of ordinary skill in the art.
Oral gels refer to the active ingredients dispersed or solubilised in a hydrophilic semi-solid matrix.
Powders for constitution refers to powder blends containing the active ingredients and suitable diluents which can be suspended e.g. in water or in juice.
Suitable diluents are substances that usually make up the major portion of the composition or dosage form. Suitable diluents include sugars such as lactose, sucrose, mannitol, and sorbitol, starches derived from wheat, corn rice, and potato, and celluloses such as microcrystalline cellulose. The amount of diluent in the composition can range from about 5 to about 95% by weight of the total composition, preferably from about 25 to about 75 weight %, and more preferably from about 30 to about 60 weight %.
The term disintegrants refers to materials added to the composition to support break apart (disintegrate) and release the pharmaceutically active ingredients of a medicament. Suitable disintegrants include starches, “cold water soluble” modified starches such as sodium carboxymethyl starch, natural and synthetic gums such as locust bean, karaya, guar, tragacanth and agar, cellulose derivatives such as methylcellulose and sodium carboxymethylcellulose, microcrystalline celluloses, and cross-linked microcrystalline celluloses such as sodium croscaramellose, alginates such as alginic acid and sodium alginate, clays such as bentonites, and effervescent mixtures. The amount of disintegrant in the composition may range from about 2 to about 20 weight % of the composition, more preferably from about 5 to about 10 weight %.
Binders are substances which bind or “glue” together powder particles and make them cohesive by forming granules, thus serving as the “adhesive” in the formulation. Binders add cohesive strength already available in the diluent or bulking agent. Suitable binders include sugars such as sucrose, starches derived from wheat corn rice and potato, natural gums such as acacia, gelatin and tragacanth, derivatives of seaweed such as alginic acid, sodium alginate and ammonium calcium alginate, cellulose materials such as methylcellulose, sodium carboxymethylcellulose and hydroxypropylmethylcellulose, polyvinylpyrrolidone, and inorganic compounds such as magnesium aluminum silicate. The amount of binder in the composition may range from about 2 to about 20 weight % of the composition, preferably from about 3 to about 10 weight %, and more preferably from about 3 to about 6 weight %.
Lubricants refer to a class of substances which are added to the dosage form to enable the tablet granules etc. after being compressed to release from the mould or die by reducing friction or wear. Suitable lubricants include metallic stearates such as magnesium stearate, calcium stearate, or potassium stearate, stearic acid, high melting point waxes, and other water soluble lubricants such as sodium chloride, sodium benzoate, sodium acetate, sodium oleate, polyethylene glycols and D,L-leucine. Lubricants are usually added at the very last step before compression, since they must be present at the surface of the granules. The amount of lubricant in the composition may range from about 0.2 to about 5 weight % of the composition, preferably from about 0.5 to about 2 weight %, and more preferably from about 0.3 to about 1.5 weight % of the composition.
Glidents are materials that prevent caking of the components of the pharmaceutical composition and improve the flow characteristics of granulate so that flow is smooth and uniform. Suitable glidents include silicon dioxide and talc. The amount of glident in the composition may range from about 0.1 to about 5 weight % of the final composition, preferably from about 0.5 to about 2 weight %.
Coloring agents are excipients that provide coloration to the composition or the dosage form. Such excipients can include food grade dyes adsorbed onto a suitable adsorbent such as clay or aluminum oxide. The amount of the coloring agent may vary from about 0.1 to about 5 weight % of the composition, preferably from about 0.1 to about 1 weight %.
Another aspect of the present invention is directed to combination therapies wherein at least one compound according to any formula (I) to (IV) is administered together with a known or commonly used drug against infectious diseases, prion diseases, immunological diseases, autoimmune diseases, bipolar and clinical disorders, cardiovascular diseases, cell proliferative diseases, diabetes, inflammation, transplant rejections, erectile dysfunction, neurodegenerative diseases and stroke. Especially preferred are combination therapies including systemic combination therapies of at least one compound of the present invention together with known or commonly used HIV drugs, antibiotics or anticancer drugs. Furthermore, the inventive compounds can also be applied in addition to chemotherapy or any other radiotherapy such as hyperthermia for cancer treatment.
In the following section, various reactions are described for the synthesis of the compounds according to general formula (I).
The urethane or thiourethane compounds of the present invention can be obtained by reaction of the corresponding amino compound with an alkyl or aryl formate as represented by the general formula R5—O—CO-LG or R5—O—CS-LG, wherein LG is a leaving group such as —Cl, —Br, —I, —N3, —O—CO—C(CH3)3 or any other suitable leaving group well known to a person skilled in the art.
The obtained urethane or thiourethane compound can further be converted to an urea or thiourea compound by consumption with a suitable amine such as primary and secondary amines. Said urea or thiourea compounds can also be obtained by conversion of the corresponding amine with an isocyanate or thioisocyanate as shown in Scheme 1.
If a residue should be bound to the six-membered ring via an oxygen atom, one suitable route may start from precursors having a keto group protected as a 1,3-dioxolane. As shown in Scheme 2, said compound having a 1,3-dioxolane residues can be converted to the free keto compound by cleavage of the 1,3-dioxolane ring with acid such as hydrochloric acid. Said deprotection reactions are well known to a skilled person. The keto compound can, for instance, further be reduced to the hydroxy compound using e.g. hydrides such as sodium borohydride. The obtained hydroxy compound can be converted to ethers when deprotonated with base such as sodium hydride, MeLi, BuLi, K—OC(CH3)3, or any other commonly used base and reacted with an electrophile represented by, for instance R14′-LG, wherein LG is a leaving group such as —Cl, —Br, —I, —O-mesylate, —O-tosylate, or any other suitable leaving group. Furthermore, the hydroxy group may be esterified by use of carboxylic acid halides, or azides, or activated esters.
Aromatization of the carbocyclic compounds such as 4,5,6,7-tetrahydro-benzo[b]furan or 4,5,6,7-tetrahydro-benzo[b]thiophene as shown in Scheme 3 can be achieved by use of MoO3/Al2O3, Pt, Pd, DDQ, SeO2, benzoquinones, or any other dehydrogenation reagents known to a person skilled in the art. Further methods for aromatization of the 6-membered carbocyclic ring are disclosed in Scheme 4.
Aromatization of the carbocyclic ring can be achieved by use of copper dichloride starting from a compound of general formula (I) wherein R8 and R9 or R10 and R11 or R12 and R13 or R16 and R17 represent together an oxygen or a 1,3-dioxolane residue or wherein R9 or R11 or R13 or R17 is a hydroxy group. As shown in Scheme 4, for instance, R16 and R17 form together a 1,3-dioxolane ring or together with carbon 6 a carbonyl group or R16 represents a hydroxy group. In this case, aromatization can be achieved with copper dichloride resulting in the corresponding 6-hydroxy-benzo[b] compound. A bromination in ortho position to the hydroxy group can be achieved by means of NBS or Br2. Said brominated product can also be obtained if the corresponding unsaturated keto compound is reacted with copper dibromide. Depending on the amount of NBS or CuBr2 used, a mono or dibromination can be obtained. The α,α-dibrominated product can be treated with a base such as potassium carbonate in order to eleminate HBr under aromatization resulting in a compound bearing a bromo and hydroxy substituent (cf. synthesis of compound A15). The bromo residue can further be substituted according to known reaction procedures.
An oxidation of the compounds of general formula (I), especially of general formula (IIa) wherein R12 and R13 represent hydrogen can be performed by the use of Cr2O72- (dochromate) in polar and protic solvents at elevated temperatures. The resulting 7-oxo-compound can then further converted to the corresponding oxime, imine, hydrazone by the use of hydroxylamine, primary amine, O-substituted hydroxylamine, hydrazine, mono or N,N-disubstituted hydrazine according to known procedures. One preferred embodiment of said conversion comprises the use of a microwave for several minutes. Ethanol or propanol or isopropanol was used as solvent and the reaction was carried out at temperature between 70° C. and 160° C., preferably 100° C. and 150° C.
A bromination of the starting material in position 7 as shown in Scheme 6 can be carried out by the use of elemental bromine and sodium acetate in acetic acid. Further substitution of the bromo residues according to known methods leads to a broad spectrum of compounds substituted in position 7.
The conversion of a cyano group to a carboxyamidine group can be achieved by the use of ammonium chloride in conjunction with AlMe3.
A carboxamide residue can be converted to the corresponding thiocarboxamide residue by the use of the commercially available Lawesson's Reagent.
The 3-sulfonic acid compounds of the present invention can be synthesized from the corresponding starting material as shown in Scheme 9 by the use of sulfonic acid chloride in methylene chloride as solvent at lower temperatures. The obtained 3-sulfonic acid compound can further be converted to the 3-sulfonic acid amide by means of, for instance, oxalyl chloride and ammonia.
The 4,7-dihydro-5H-thieno[2,3-c]pyran compounds according to the present invention are obtainable by different synthetic routes. One route, which leads to 4,7-dihydro-5H-thieno[2,3-c]pyran derivatives starts with the reaction of tetrahydro-pyran-4-one or a correspondingly substituted derivative thereof with an cyano-actetate ester under acidic or basic conditions, preferably under acidic conditions, and under elimination of water and subsequent reaction of the reaction product with sulfur in the presence of an organic base to give a corresponding 2-amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ester derivative.
As a next step, the amino group in the thus obtained 2-amino4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ester derivative can be acylated to give a corresponding 2-carbonylamino compound. As an acylation reagent a carboxylic acid chloride is preferably used. This reaction can optionally be carried out in the presence of a base such as an tertiary amide, preferably NEt(iPr)2.
Other suitable reactions to obtain the secondary carboxylic acid amides can be used, for instance reaction of the amino group with a carboxylic acid and a coupling-agent as used in peptide chemistry, such as HOBT,HOOBT,HBTU or HOAt.
Alternatively, if instead of the acyl group a sulfonyl group is to be attached to the amino group in 2-position, the 2-amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ester derivative can be reacted with a sulfonyl chloride compound to give a corresponding 2-sulfanylamino derivative.
The thus obtained compounds can then optionally be reacted with bromine in the presence of an organic acid, preferably acetic acid, to substitute one hydrogen in 7-position of the heterocyclic nucleus by a hydroxyl group.
The above described 3-carboxylic acid ester derivative compounds can then be reacted in a subsequent reaction step with an alkali metal amide, such as LiNH2 or NaNH2, in a polar solvent, which is essentially inert to the alkali metal amide, to give the corresponding 3-carboxylic acid amide derivative. This reaction is preferably carried out under the exclusion of moisture and optionally under an inert atmosphere. The application of lithium amide instead of sodium amide results in higher yields and purer products.
To prepare the corresponding 4,7-dihydro-5H-thieno[2,3-c]pyran derivatives in which a sulfonamide is attached in 3-position, in a first step, 4,7-dihydro-5H-thieno[2,3-c]pyran-2-amine can be acylated, preferably using a carboxylic acid chloride to give the corresponding 2-carbonyl-amino derivative. This compound can then be reacted with sulfurylchloride, preferably under an inert atmosphere and subsequently with ammonia to give the 3-sulfonamide compound.
If the compounds used to synthesise the compounds according to the present invention contain —NH, —SH or —OH functional groups which potentially interfere with the desired reaction, these may of course be protected with suitable protective groups, which can later on be removed from the respective compounds.
To obtain those analogues of the 4,7-dihydro-5H-thieno[2,3-c]pyran derivatives in which the S-atom in the 5-membered ring of the heterocyclic nucleus is substituted either by NR4′ or O, the following synthetic approach can be utilized, which is partially based on a method described in Hauser, C. R., Hoffenberg, D. S.; J. Org. Chem. 1955, 20, 1448-1453.
To obtain the O-analogue compounds, the amino group in 2-position of a corresponding 2-amino-3-cyano-4,7-dihydro-5H-furo[2,3c]pyrane derivative can be acylated in a first reaction step, using the acylation reaction described above with reference to the acylation of the 2-amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ester derivatives, i.e. preferably using a carboxylic acid chloride as an acylation agent, obtionally in the presence of a tertiary amine base such as NEt(iPr)2. Similarly, to obtain the NR4′-analogue compounds, a corresponding 2-amino-3-cyano-4,7-dihydro-5H-pyrrolo[2,3-c]pyrane derivative is acylated in the above described manner.
The respective 2-carbonyl-amino derivatives obtained by this acylation can then be reacted with boron trifluoride-acetic acid complex [BF3.(HOAc)2] and subsequently treated with an aqueous alkali metal hydroxide solution, such as sodium hydroxide, to convert the cyano group in 3-position of the heterocyclic nucleus into the carboxamide group.
In a further aspect of the present invention, the invention is directed at a method for amidation of an carboxylic acid ester to give the corresponding primary carboxylic acid amide. This amidation comprises the step of reacting an carboxylic acid ester with an alkali metal amide in the presence of a polar solvent, which is essentially inert against the alkali metal amides. Preferably, the molar ratio of carboxylic acid ester to alkali metal amide lies in the range of 1:1 to 1:15, more preferably in the range of 1:5 to 1:14, and most preferably in the range of 1:9 to 1:13.
In a preferred embodiment of the method of the present invention, the alkali metal amide is LiNH2 or NaNH2, and preferably is LiNH2. The solvent is preferably absolute ether or absolute tetrahydrofurane, preferably tetrahydrofurane, and the reaction is preferably carried out under the exclusion of moisture. Preferably, the reaction is carried out at a temperature of 15° C. to 35° C., preferably at 25° C. It is furthermore preferred that the reaction duration lies in the range of from 40 to 80 hours, preferably from 45 to 75 hours.
According to the following inventive procedure various amides can be synthesized:
wherein
X2 represents O or S, and
R1, R3, R4, Y1-Y4 and X1 have the meanings as defined in claim 1 or any part of the description.
In a preferred embodiment of said method of the present invention, the carboxylic acid ester is a compound according to the following general formula (D):
which is amidated to give the primary carboxylic acid amide according to formula (E),
wherein in formulas (D) and (E)
X1 is selected from S, O, or NR1, and R1 is selected from H, substituted or unsubstituted C1-C6-alkyl,
R2 is linear or branched C1-C6 alkyl or aryl and preferably is methyl, ethyl, phenyl or benzyl,
R4 is selected from H, —C(═X2)R5 and —SO2R5,
R8 is H and R9 is selected from H, substituted or unsubstituted C1-C6-alkyl
R10 is selected from H, substituted or unsubstituted C1-C6-alkyl, C1-C6-alkoxy, or OH
R11 is selected from H and substituted or unsubstituted C1-C6-alkyl
R12 is selected from H and substituted or unsubstituted C1-C6-alkyl, C1-C6-alkoxy, or
OH, and
R13 is selected from H or substituted or unsubstituted C1-C6-alkyl,
and stereoisomeric and regioisomeric forms and pharmaceutically acceptable salts of these compounds.
In a further preferred embodiment of the method of the present invention, in general formulas (D) and (E)
X1 is S
R2 is methyl or ethyl,
R4 is —C(═O)R5 and R5 is selected from methyl, ethyl, propyl, butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, C1-C10-cycloalkyles substituted by at least one methyl or carboxyl group, phenyl, furanyl, thienyl, pyrrolyl, pyridyl, pyrrolidinyl, piperidinyl, tetrahydrofuranyl, ethenyl, cis-prop-1-enyl, trans-prop-1-enyl, cis-prop-2-enyl, trans-prop-2-enyl, but-1-enyl, cis-but-2-enyl, trans-but-2-enyl, but-3-enyl, prop-1-inyl, prop-2-inyl, but-1-inyl, but-2-inyl, but-3-inyl or adamantyl,
R8 is H and R9 is selected from H, substituted or unsubstituted C1-C6-alkyl,
R10 is selected from H, substituted or unsubstituted C1-C6-alkyl, C1-C6-alkoxy, or OH,
R11 is selected from H and substituted or unsubstituted C1-C6-alkyl,
R12 is selected from H and substituted or unsubstituted C1-C6-alkyl, C1-C6-alkoxy, or OH, and
R13 is selected from H or substituted or unsubstituted C1-C6-alkyl.
According to one preferred embodiment of the method of the present invention, the compound according to the general formula (E) is obtained by the following reaction sequence:
Step I:
Step II: Acylation of the —NH2 group in 2-position in compound (C) with R5C(═O)LG, wherein LG represents a suitable leaving group, preferably a halogen such as F, Cl, Br or I, most preferably Cl, to give compound (D):
and,
Step III: Amidation of compound (d) as outlined above by means of a sodium or lithium amide,
It is preferred that in Step I the reaction of compound (B) with the cyano-acetate ester is carried out in a nonpolar solvent, preferably benzene, with the addition of a mixture of ammonium acetate and acetic acid in a molar ratio of greater than 1, preferably in the range from 0.5:1 to 0.8 to 1, and preferably at a temperature in the range of 50 to 100° C., preferably between 70 to 90° C., preferably under removal of water formed in the reaction, and preferably for a duration of 2 to 4 hours.
Furthermore, in a preferred embodiment of the present invention, in Step I the reaction product of the reaction of compound (B) with the cyano-acetate ester is reacted with the S8 in a protic solvent, preferably EtOH, S8 being added at least in aquimolar quantities, preferably in an excess of up to 1,5, more preferably of up to 1,2, in the presence of a amine base, preferably morpholine, at reaction temperature of between 25 to 65° C., preferably between 40 and 60° C., and preferably for a duration of 2 to 6 hours.
Analytical Methods:
LC/MS data were obtained using a Waters Micromass ZQ instrument with atmospheric pressure chemical ionisation or electrospray ionisation under the conditions described below.
MS Detection:
Scan range for MS Data (m/z)
Start (m/z) 100
End (m/z) 600
With +ve/−ve switching
Ionisation is either electrospray or APCI dependent on compound types.
Standard Prep HPLC Conditions (Method 1)
Standard LC-MS Conditions
UV Detection via Waters 2996 PDA
For purity assessments the wavelengths at 21 5, 254 and 310 nm were extracted from the PDA data and an average purity was calculated from the peak areas.
All reagents were obtained commercially and used directly. DMF and THF were dried over 4 Å molecular sieves (Fluka). Column chromatography employed Silica Gel 60 (Merck). TLC was carried out using pre-coated aluminium sheets Silica gel 60 F254 (Merck).
Standard Conditions for Flash Chromatography
Flash chromatography was done using a SiO2-column and the following solvents: cyclohexane (cHex), ethyl acetate (EtOAc), dichloromethane (DCM), methanol (MeOH).
To a 1:1:1 mixture of the corresponding cyclic ketone, acetonitrile derivative and sulfur in ethanol, morpholine (1.1 eq.) was added dropwise and the mixture heated at 45° C. for 4 h. The sulfur dissolved slowly and after approximately 2 h a yellow precipitate was formed. For complete precipitation the mixture was stored at 4° C. overnight. The solid was filtered, washed with water and re-crystallized from ethanol to give pale-white to yellow crystals of the product.
As an example, the following intermediates were prepared:
Mp.: 186-187° C. (ethanol).
Mass calc. for C9H12N2OS: 196.27, found (pos. mode) 197.26.
Mp.: 157-158° C. (ethanol).
Mass calc. for C11H14N2O3S: 254.31, found (pos. mode) 255.4, found (neg. mode) 253.3.
Mp.: 212-214° C. (ethanol).
Mass calc. for C14H18N2O3S: 226.30, found (pos. mode) 249.1 [M+Na+], found (neg. mode) 225.2.
Mass calc. for C14H18N2O3S: 226.30, found (pos. mode) 227.1, found (neg. mode) 225.4.
Mass calc. for C8H10N2O2S: 198.25, found (pos. mode) 199.3, found (neg. mode) 197.2.
Mp.: 140-145° C. (ethanol).
Mass calc. for C13H20ON2OS: 252.38, found (pos. mode) 253.4, found (neg. mode) 251.4.
Mass calc. for C10H14N2O2S: 226.30, found (pos. mode) 227.2, found (neg. mode) 225.2.
Mp.: 201-204° C. (ethanol).
Mass calc. for C12H16N2O3S: 268.34, found (pos. mode) 269.1, found (neg. mode) 267.2.
A suspension of 196 mg (1.00 mmol) of 2-amino-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxamide, 100 μl (1.10 mmol) of cyclopropanecarboxylic acid chloride and 218 μl (1.25 mmol) of diisopropylethylamine in 3 ml toluene was heated at 90° C. for 4 h. After cooling at room temperature the mixture was diluted with 20 ml EtOAc and washed with 15 ml of 0.5 M HCl-solution. The water phase was extracted twice with 15 ml EtOAc and CH2Cl2. The combined organic layers were dried over Na2SO4. Evaporation of the solvent and recrystallization of the residue from ethanol gave the product as a light orange powder in 78% yield.
Mp.: 216-217° C. (ethanol).
Mass calc. for C13H16N2O2S: 264.34, found (neg. mode) 263.23.
0.5 mmol of the corresponding carboxylic acid and 5 drops of abs. DMF were dissolved in 10 ml of abs. THF or abs. DCM under nitrogen atmosphere. 0.56 mmol (1.1 equivalents) of oxalyl chloride were slowly added via syringe at room temperature, and the solution was stirred for 2 h. The solvents were evaporated, and the residue (usually approximately 0.5 ml) was re-dissolved in 5 ml of abs. THF under nitrogen atmosphere. For some compounds the commercially available acid chloride was used instead. 0.56 mmol (1.1 equivalents) of DIPEA were added via syringe, followed by a suspension of 0.5 mmol (1 equivalent) of 2-amino-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxamide or 2-amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxamide in 5 ml of abs. THF. The suspension was stirred at room temperature for 2-24 h. 50 ml of saturated NH4Cl solution were added, and the mixture was extracted three times with EtOAc. The combined organic layers were washed with water and dried over Na2SO4. Filtration and evaporation of the solvent gave the crude product, which was usually purified by recrystallization from hot ethanol. This reaction can be run with pyridine as solvent and base and also in a 1:1 mixture of THF-pyridine or DMSO-pyridine. In the latter two cases a work-up with cold 1N HCl is necessary.
According to this procedure the following compounds were prepared:
D127: Mp.: 198-199° C. (ethanol).
Mass calc. for C16H20N2O4S: 336.41, found (pos. mode) 337.2, found (neg. mode) 335.2.
D97: Mp.: 199-200° C. (ethanol).
Mass calc. for C15H18N2O4S: 322.39, found (pos. mode) 323.3, found (neg. mode) 321.2.
B156: Mp.: 223-225° C. (ethanol).
Mass calc. for C14H18N2O3S: 294.38, found (pos. mode) 317.4 [M+Na+], found (neg. mode) 293.4.
B71: Mp.: 195-196° C. (ethanol).
Mass calc. for C14H18N2O3S: 294.38, found (pos. mode) 317.4 [M+Na+], found (neg. mode) 293.4.
D96: Mp.: 241° C. (ethanol, decomposition).
Mass calc. for C12H14N2O3S: 266.32, found (pos. mode) 289.3 [M+Na+], found (neg. mode) 265.4.
D95: Mp.: 230-231° C. (ethanol, decomposition).
Mass calc. for C17H24N2O2S: 320.46, found (pos. mode) 343.4 [M+Na+], found (neg. mode) 319.4.
D201: Mp.: 185° C. (ethanol).
Mass calc. for C14H18N2O3S: 294.38, found (pos. mode) 295.1.
D146: Mp.: 211-213° C. (ethanol).
Mass calc. for C16H20N2O4S: 336.41, found (pos. mode) 337.1, found (neg. mode) 335.2.
B174: Mp.: 106-108° C.
Mass calc. for C16H20N2O4S: 336.41, found (pos. mode) 337.2.
B40: Mass calc. for C20H22N2O4S: 386.47, found (pos. mode) 387.1, found (neg. mode) 385.2.
B172: Mp.: 180-181° C.
Mass calc. for C14H18N2O4S: 310.37, found (pos. mode) 311.2 found (neg. mode) 309.1.
D94: Mp.: 207-209° C. (EtOH).
Mass calc. for C14H18N2O2S: 278.37, found (neg. mode) 277.21.
B221: Mass calc. for C14H18N2O3S: 294.37, found (pos. mode) 295.13, found (neg. mode) 293.16.
B175: Mp.: 84-94° C. (flash-column chromatography on silica with cHex/EtOAc 1:1).
Mass calc. for C20H30N2O3S: 378.54, found (neg. mode) 377.33.
B42: Mass calc. for C16H18Cl2N2O3S: 388.04, found (pos. mode) 389.05, found (neg. mode) 387.08.
B194: Mp.: 214-217° C. (EtOH; decomposition).
Mass calc. for C14H16Cl2N2O3S: 362.03, found (pos. mode) 363.04, found (neg. mode) 361.09.
Mp.: 177-179° C.
Mass calc. for C16H17NO4S: 319.38, found (pos. mode) 320.24.
To a solution of 210 mg (1 mmol) of 2-Amino-6-methyl-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide in 2 mL pyridine, 69 mg (0.5 mmol) of PCl3 were added at −25° C. After 20 minutes 1 mmol of (4-nitro-phenyl)-acetic acid was added to the solution at −25° C., and the stirring continued at room temperature for 12 h. The reaction mixture was evaporated to dryness, dissolved in ethyl acetate, washed twice with 15 ml water, 15 ml 5% NaHCO3, and 15 ml saturated NaCl solution. The combined organic layers were dried over Na2SO4 and evaporated to dryness. After washing with n-hexane and isopropanol a solid product was obtained in 63% yield.
The following compounds were prepared by this method:
260 mg (1 mmol) 2-amino-4,7-dihydro-5H-spiro[1-benzothiophene-6,2′-[1,3]dioxolane]-3-carboxamide was stirred in 3 ml dry pyridine and 1.1 mmol of the corresponding isocyanate was added at room temperature and stirred overnight. Then 3 ml of distilled water was added and the precipitated crystals were filtered and dried in vacuum.
The following compounds were obtained according to this method:
To a suspension of 100 mg (0.51 mmol) 2-amino-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxamide in 2.5 ml ethanol, 56 mg (0.56 mmol) of cyclopropylthioisocyanate were added dropwise. The mixture was heated to reflux and became a solution. After approximately 1 h a colourless precipitate was formed. After 2 h of reflux the mixture was cooled and stored at 4° C. overnight. The solid was filtered and washed with cold ethanol to yield the colourless product in 57% yield. ethanol can be replaced by a 1:1 mixture of DMSO abs. and pyridine abs. In this case the reaction is run at 90-100° C.
Mp.: 206-207° C. (decomposition, ethanol).
Mass calc. for C13H17N3OS2: 295.42, found (neg. mode) 294.30, (pos. mode) 296.24.
According to this procedure the following compounds were prepared:
A solution of 1.0 g (3.9 mmol) 2-amino-4,7-dihydro-5H-spiro[1-benzothiophene-6,2′-[1,3]dioxolane]-3-carboxamide in 20 ml THF was treated dropwise with 425 μl (3.9 mmol) phenyl isocyanate at 0° C. and stirred at 0° C. for 30 min. The mixture was allowed to come to room temperature and stirred for 3 h during which time a colourless precipitate was formed. The solid was filtered and washed with a small amount of cold THF and water. Addition of some water to the mother liquid afforded a second batch of the product as a colourless precipitate, which was filtered and washed with water and methanol. The combined product was obtained in 80% yield. THF can be replaced by a 1:1 mixture of DMSO abs. and pyridine abs. or by 1,4-dioxane (with 0.5 eq of DMAP). If the product does not precipitate, then the reaction mixture is poured onto ice-water and the pH is adjusted to pH 2 with diluted HCl. The precipitate is filtered, washed with water and dried.
Mp.: 212-213° C. (THF).
Mass calc. for C18H19N3O4S: 373.43, found (pos. mode) 374.1, found (neg. mode) 372.1.
According to this procedure the following compounds were prepared:
D178: Mp.: 219-220° C. (THF).
Mass calc. for C18H18FN3O4S: 391.42, found (pos. mode) 392.1, found (neg. mode) 390.2.
D176: Mp.: 198-199° C. (THF).
Mass calc. for C19H21N3O5S: 403.46, found (pos. mode) 404.2, found (neg. mode) 402.2.
D175: Mp.: 182-183° C. (THF).
Mass calc. for C18H18IN3O4S: 499.33, found (pos. mode) 500.0, found (neg. mode) 498.1.
A mixture of 7.1 g (20.73 mmol) of (3-carbamoyl-6-hydroxy-benzo[b]thiophen-2-yl)-carbamic acid benzyl ester, 13.3 g (211 mmol) of ammonium formate, and 7.1 g of 5% palladium on carbon in 900 ml of methanol was stirred under argon for 24 h. The mixture was filtered through Celite and the filtrate concentrated under reduced pressure. The residue was treated with water, the precipitate was filtered off and washed with water and diethyl ether affording 2.15 g (49.9%) of the title compound.
HPLC method B; retention time: 1.83; M−: 207.03
The mixture of 1.66 g (8. mmol) of 2-amino-6-hydroxy-benzo[b]thiophene-3-carboxylic acid amide, 2.2 g (16 mmol) potassium carbonate, 50 ml of acetone and 1.1 ml (1.29 g 8.4 mmol) diethylsulfate was stirred at reflux temperature for 24 hours. The mixture was concentrated under reduced pressure. The residue was treated with 20 ml water and 10 ml of ethylacetate. The insoluble material was filtered off, washed with water, a little ethylacetate and dried affording 1.66 g (yield: 61%) of the title compound as a white solid.
HPLC method B; retention time: 2.84; M−: 235.09
NMR (300 MHz, DMSO-d6 ppm): 7.57 (d, 1H), 7.46 (s, 2H), 7.25 (s, 1H), 6.85 (m, 3H), 4.00 (q, 2H), 1.31 (t, 3H)
The mixture of 14 ml abs. dioxane and 7 ml abs. THF was cooled down to 0° C. and phosgene was bubbled through the solution for 30 min. 2.53 mmol of the amino derivative were dissolved in 5 ml abs. dioxane and 2 ml abs. triethylamine and added slowly to the cold mixture. After stirring for 1 h the mixture was evaporated and 4 ml abs. DMSO was added to the isocyanate compound. 0.5 g (2.53 mmol) 2-Amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide was dissolved in 8 ml abs. DMSO and 4 ml abs. pyridine and added to the isocyanate-solution. After 2 h stirring at room temperature the mixture was poured on 1N HCl, filtered off, washed with water and dried.
According to this procedure the following compounds were prepared:
Nicotinyl chloride was dissolved in acetone, cooled to −10° C. and an aqueous solution of sodium azide was added. After 30 minutes the reaction mixture was diluted with cold distilled water and extracted 3 times with cold toluene. The organic layer was dried on Na2SO4 at 4° C. and filtered. The filtrate was added dropwise into refluxing toluene while stirring and refluxed for one hour. Toluene was evaporated and pyridine-3-isocyanate was obtained as a yellow oil and it was condensed with the corresponding amine as described in Procedure 1. The acyl chloride derivative can be replaced also by a mixed anhydride obtained by reaction of a carboxylic acid with a 10% excess of isobutyl chloroformate and triethylamine.
According to this procedure the following compounds were prepared:
1 mmol heteroarylamine was dissolved in 2.5 ml THF and 0.33 mmol triphosgene were added at 0° C. to the stirred solution under argon atmosphere. After 5 minutes, 3 mmol triethylamine was added dropwise while keeping the temperature at 0° C. for an additional 5 minutes, then 1 mmol 2-amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide in 2.5 ml DMSO was added and the reaction mixture was allowed to reach room temperature and stirred for 2 h. The reaction mixture was poured onto ice-water and the precipitated product was filtered off and recrystallized from ethanol-water.
According to this procedure the following compounds were prepared:
1.2 g (3.7 mmol) 2-(cyclopropanecarbonyl-amino)-4,7-dihydro-5H-spiro[1-benzothiophene-6,2′-[1,3]dioxolane]-3-carboxamide in 20 ml THF and 10 ml 1 M HCl solution was heated to reflux for 6 h under nitrogen atmosphere. Sat. NaHCO3 solution was added to give a neutral water phase and the mixture was extracted four times with 50 ml EtOAc. The combined organic layers were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the product as a light red powder in 85% yield. Recrystallization from ethanol afforded the product as a colourless solid.
Mp.: 201-202° C. (ethanol).
Mass calc. for C13H14N2O3S: 278.33, found (pos. mode) 279.1, found (neg. mode) 277.2.
According to this procedure the following compounds were prepared:
D204:
Mass calc. for C14H16N2O3S: 292.36, found (pos. mode) 293.2, found (neg. mode) 291.2.
D106: Mp.: 210-211° C.
Mass calc. for C17H17N3O4S: 359.41, found (pos. mode) 360.2, found (neg. mode) 358.2.
D105: Mp.: 221-222° C.
Mass calc. for C16H14IN3O3S: 455.28, found (pos. mode) 456.1, found (neg. mode) 454.1.
The 1,3-dioxolane derivative (2 mmol) was dissolved in abs. dichloromethane (5 ml) and trifluoroacetic acid (2 ml) was added. After stirring at room temperature for 24 h, the solution was concentrated under vacuum, and the solid material was filtered and washed with n-hexane (2×20 ml).
According to this procedure the following compounds were prepared:
113 mg (0.41 mmol) of 2-(cyclopropanecarbonyl-amino)-6-oxo-4,5,6,7-tetrahydro-benzo[b]-thiophene-3-carboxamide was suspended in 4 ml methanol, and 19 mg (0.49 mmol) of NaBH4 were added portion-wise. The mixture was stirred at room temperature for 3 h. After addition of a small amount of water, the solvent was evaporated and the residue taken up in 10 ml sat. NH4Cl solution and 20 ml EtOAc. Extraction with EtOAc (4×30 ml), washing of the combined organic layers with brine, drying over Na2SO4 and evaporation of the solvent gave the crude product, which, upon recrystallization from ethanol, afforded the product as colourless crystals.
Mp.: 238-239° C. (ethanol).
Mass calc. for C13H16N2O3S: 280.35, found (pos. mode) 281.29, found (neg. mode) 279.23.
According to this general method the following compounds were prepared:
D202: Mp.: 224-230° C. (ethanol).
Mass calc. for C14H18N2O3S: 294.38, found (pos. mode) 295.2, found (neg. mode) 293.2.
D141: Mp.: 180-181° C. (ethanol/methanol, decomposition).
Mass calc. for C17H19N3O4S: 361.42, found (pos. mode) 362.1, found (neg. mode) 360.1.
D140: Mp.: 201-202° C. (ethanol/methanol).
Mass calc. for C16H16IN3O3S: 457.29, found (pos. mode) 458.0, found (neg. mode) 456.1.
A suspension of 500 mg (1.55 mmol) 2-(cyclopropanecarbonyl-amino)-4,7-dihydro-5H-spiro[1-benzothiophene-6,2′-[1,3]dioxolane]-3-carboxamide in 10 ml CH3CN was treated with 528 mg (3.10 mmol) CuCl2.2 H2O at room temperature. The mixture turned brown immediately and was stirred at room temperature for 2 h. The solvent was evaporated and the residue taken up in 20 ml 0.5 M HCl solution and 40 ml. EtOAc. The mixture was extracted with EtOAc. The combined organic layers were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by flash column chromatography on silica gel (eluent cHex/EtOAc 1:2) to afford a slightly purple powder in 79% yield. Further purification by preparative HPLC (Method 1) or recrystallization from ethanol afforded the product as a colourless solid.
For the work-up it is also possible to filter off the purple solid directly after the reaction. After washing the solid with 10 ml acetonitrile, 100 ml 0.3 M HCl solution and finally twice with 20 ml THF, and drying it, a pure product was obtained which could be used in further steps without additional purification.
Mp.: 278-281° C. (CH3CN/H2O).
Mass calc. for C13H12N2O3S: 276.31, found (pos. mode) 277.2, found (neg. mode) 275.3.
According to this procedure the following compounds were prepared:
A5: Mp.: 220-221° C. (ethanol).
Mass calc. for C14H14N2O3S: 290.34, found (pos. mode) 291.2, found (neg. mode) 289.2.
A12: Mp.: >290° C. (CH3CN/H2O).
Mass calc. for C16H13N3O3S: 327.36, found (pos. mode) 328.1.
A24: Mp.: >280° C. (CH3CN/H2O).
Mass calc. for C16H11ClFN3O3S: 379.80, found (pos. mode) 380.1 & 382.1, found (neg. mode) 378.1 & 380.1.
A11: Mass calc. for C17H15N3O4S: 357.39, found (pos. mode) 358.1, found (neg. mode) 356.1.
A33: Mp.: 226-227° C. (CH3CN/H2O).
Mass calc. for C16H12IN3O3S: 453.26, found (pos. mode) 454.0 & 456.0, found (neg. mode) 452.1 & 454.1.
A suspension of 100 mg (0.36 mmol) (6-R,S)-2-(cyclopropanecarbonyl-amino)-6-hydroxy-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxamide and 12 mg (0.36 mmol) sulfur in 1.5 ml dimethyl phthalate was heated in the microwave at 245° C. for 20 min. The resulting brown solution was evaporated and the residue purified by flash column chromatography on silica gel (eluent cHex/EtOAc 1:2) afford a colourless solid in 40% yield.
Mass calc. for C13H12N2O3S: 276.31, found (pos. mode) 277.2, found (neg. mode) 275.3.
100 mg (0.25 mmol) of 6-Oxo-2-[3-(4-trifluoromethyl-phenyl)-ureido]-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide and 80 mg palladium on charcoal (10%) were suspended in the mixture of 18 ml glacial acetic acid and 0.5 ml water. The mixture was stirred at 40° C. for 48 h, then filtered, and the solution was evaporated to dryness to give the title compound in 89% yield.
According to this procedure the following compounds were prepared:
Benzoquinone (1.5 mmol) was added to a solution of 2-[3-(4-bromo-phenyl)-ureido]-6-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide in acetic acid. The mixture was stirred overnight at 50° C. The solvent was evaporated, the residue was dissolved in ethyl acetate and washed several times with saturated Na2SO3, brine and dried over Na2SO4. Evaporation of the solvent gave a solid, which was crystallized from isopropanol.
Yield: 60%
The following compounds were prepared by this method:
DDQ (2 mmol) and a catalytic amount of p-toluenesulfonic acid were added to a solution of 3-carbamoyl-2-(cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]thiophene-6-carboxylic acid ethyl ester in acetic acid and the mixture was stirred for 2 h at 100° C. The solvent was evaporated and the residue was dissolved in ethyl acetate and washed several times with saturated Na2SO3 and brine dried over Na2SO4. Evaporation of the solvent gave the product (Yield: 70%).
The following compounds were prepared by this method:
300 mg (0.80 mmol) 2-[(anilinocarbonyl)amino]-4,7-dihydro-5H-spiro[1-benzothiophene-6,2′-[1,3]dioxolane]-3-carboxamide was dissolved in 15 ml THF and 5 ml 1 M HCl solution and heated to reflux (oil bath 80° C.) overnight. The mixture was stored in the fridge overnight and the precipitated 6-keto product filtered and washed with water and some THF. After drying of the crude 6-keto compound under fine vacuum overnight (crude yield: 76%), it was reduced as described in general method 4.
Mp.: 153-154° C. (ethanol/methanol).
Mass calc. for C16H17N3O3S: 331.40, found (pos. mode) 332.1, found (neg. mode) 330.2.
According to this general method the following compound was prepared:
D142: Mp.: 180-181° C. (ethanol/methanol).
Mass calc. for C16H16FN3O3S: 349.39, found (pos. mode) 350.1, found (neg. mode) 348.2.
53 mg (0.24 mmol) CuBr2 was added to a solution of 60 mg (0.22 mmol) 2-(cyclopropanecarbonyl-amino)-6-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxamide in 1 ml CH3CN, 1 ml CHCl3 and 20 μl EtOH. The mixture was stirred at room temperature for 4 h. 10 ml water were added and the mixture extracted three times with 20 ml EtOAc. The combined organic layers were washed with brine, dried over Na2SO4 and the solvent was evaporated. Separation by preparative HPLC (Method 1) afforded the title compound as a light brown solid in 5% yield.
Mass calc. for C15H16N2O3S: 304.36, found (pos. mode) 305.2, found (neg. mode) 303.2.
According to this general method the following compound was prepared:
A23: Mp.: >260° C. (CH3CN/H2O).
Mass calc. for C19H19N3O4S: 385.44, found (pos. mode) 386.2, found (neg. mode) 384.2.
To a suspension of 19 mg (0.47 mmol) NaH (60% dispersion in mineral oil) in 2 ml DMF was added a solution of 100 mg (0.36 mmol) 2-(cyclopropanecarbonyl-amino)-6-hydroxy-benzo[b]thiophene-3-carboxamide in 2 ml DMF at 0° C. The mixture was stirred at room temperature for 1 h, cooled again to 0° C. and treated with 30 μl (0.47 mmol) methyl iodide. After stirring overnight at room temperature, the mixture was treated with 20 ml sat. NH4Cl-solution and extracted three times with 30 ml EtOAc. The combined organic layers were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by flash column chromatography on silica gel (eluent cHex/EtOAc 1:1) to afford a reddish foam in 32% yield.
Mass calc. for C14H14N2O3S: 290.34, found (pos. mode) 291.1, found (neg. mode) 289.1.
According to this general method the following compounds were prepared:
A34: Mp.: 190-192° C. (CH3CN/H2O).
Mass calc. for C20H25N3O3S: 387.50, found (pos. mode) 388.2, found (neg. mode) 386.2.
A19: Mp.: 194-195° C. (CH3CN/H2O).
Mass calc. for C16H19N3O3S: 333.41, found (pos. mode) 334.2.
A25: Mp.: 172-173° C. (ethanol).
Mass calc. for C18H21N3O4S: 375.45, found (pos. mode) 376.2, found (neg. mode) 374.2.
A29: Mp.: 200-202° C. (EtOAc).
Mass calc. for C16H16N2O5S: 348.38, found (pos. mode) 349.1, found (neg. mode) 347.2.
A21: Mp.: 180-183° C. (H2O/CH3CN).
Mass calc. for C20H24N2O5S: 404.49, found (pos. mode) 405.2, found (neg. mode) 403.3.
D144: In this example 3.26 equivalents of NaH (60% dispersion in mineral oil) and dimethyl sulfate as the electrophile were used.
Mass calc. for C14H8N2O3S: 294.38, found (pos. mode) 295.2, found (neg. mode) 293.2.
A10: Mass calc. for C20H24ClN3O3S: 421.95, found (pos. mode) 422.2 & 424.1, found (neg. mode) 420.2 & 422.2.
The above outlined reaction scheme is exemplified by the synthesis of A49 and A50.
0.28 g (0.10 mmol) 2-(Cyclopropanecarbonyl-amino)-6-hydroxy-benzo[b]thiophene-3-carboxylic acid amide and 0.40 g (3.00 mmol) N-chloro-succinimide was stirred at room temperature between 4 to 24 hours in 15 ml acetonitrile. The reaction mixture was diluted with 15 ml water, the solid was filtered off, and was washed three times with 5 ml water. The crude product was refluxed in 10 ml methanol for half an hour, was cooled to room temperature, and was filtered off to give pure product in 80% yield.
Mp.: 240-245° C.
Mass calc. for C13H10Cl2N2O3S: 345.21.
NMR, δ (ppm): 11.83 (broad s, 1H), 10.22 (broad s, 1H), 7.86 (s, 1H), 7.80 (s, 2H), 2.05 (m, 1H), 0.94 (m, 4H).
Compound A120 was prepared according to this reaction procedure.
Yield of A120: 0.22 g (70%)
Mp.: 235-240° C.
NMR, δ (ppm): 11.24 (broad s, 1H), 10.24 (broad s, 1H), 7.88 (s, 1H), 7.78 (broad s, 2H), 4.23 (m, 2H), 1.27 (t, J=7.08 Hz, 3H).
0.18 g (0.50 mmol) 6-hydroxy-5,7-dichloro-2-(cyclopropanecarbonyl-amino)-benzo[b]thiophene-3-carboxylic acid amide was treated with 0.12 g (1.00 mmol) potassium-tert-butoxide at room temperature in 20 ml absolute dimethyl-formamide. After stirring at room temperature for one hour 0.06 mmol alkyl-halide derivative was added into the reaction mixture, and it was heated at 80° C. for four hours. Then the solvent was evaporated under reduced pressure, the residue was titurated with 20 ml water, and extracted three times with 20 ml ethyl-acetate. The combined organics were washed with 20 ml brine and dried over magnesium-sulphate. Evaporation of the solvent gave the crude product, which was recrystallised from ethanol afforded the pure product.
According to this procedure the following compounds were prepared:
0.17 g (0.34 mmol) {2-[3-Carbamoyl-5,7-dichloro-2-(cyclopropanecarbonyl-amino)-benzo[b]thiophen-6-yloxy]-ethyl}-carbamic acid tert-butyl ester was dissolved in 20 ml methanol, and 1.0 ml ethyl acetate saturated with hydrochloric acid was added into the reaction mixture. After stirring for two hours at room temperature, the solvent was evaporated. The crude product was recrystallised from 5 ml ethanol-(2-propanol) 1-1 mixture to give pure product.
Yield: 65%
Mass calc. for C15H15Cl2N3O3S: 388.28.
NMR, δ (ppm): 11.89 (broad s, 1H), 8.28 (broad s, 3H), 7.97 (s, 1H), 7.88 (s, 2H), 4.24 (t, J=5.22 Hz, 2H), 3.27 (m, 2H), 2.09 (m, 1H), 0.98 (m, 4H).
60 mg (0.17 mmol) [3-carbamoyl-2-(cyclopropanecarbonyl-amino)-benzo[b]thiophen-6-yloxy]-acetic acid methyl ester was dissolved in 2 ml dioxane and treated with 40 μl (0.66 mmol) ethanolamine. Under stirring 20 mg (0.52 mmol) NaH (60% dispersion in mineral oil) were added. The microwave tube was sealed and heated in the microwave at 100° C. for 360 s. Since the conversion was not complete, the reaction was allowed to stir at room temperature for 5 days. The mixture was treated with 15 ml sat. NH4Cl solution and extracted four times with 20 ml EtOAc. The combined organic layers were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by preparative HPLC (Method 1) to afford the product as a colourless solid in 25% yield.
Mp.: 220-222° C. (H2O/CH3CN).
Mass calc. for C17H19N3O5S: 377.42, found (pos. mode) 378.2, found (neg. mode) 376.2.
According to this general method the following compounds were prepared:
A7: Mp.: 249-251° C. (H2O/CH3CN).
Mass calc. for C16H17N3O4S: 347.40, found (pos. mode) 348.2, found (neg. mode) 346.2.
D79: Mp.: 252-254° C. (CH3CN/H2O).
Mass calc. for C16H21N3O4S: 351.43, found (pos. mode) 352.2, found (neg. mode) 350.2.
According to this procedure the following compounds were prepared:
A solution of 8.4 g (28.5 mmol) potassium dichromate in 12 ml water at 60° C. was slowly added to 2.5 g (9.5 mmol) 2-(cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide dissolved in 20 ml acetic acid at 60° C. and the reaction was then stirred at 80° C. for 7 h. After the starting material disappeared, the reaction mixture was poured on ice (80 g), and the aqueous layer was extracted three times with EtOAc (250 ml). Evaporation to dryness and recrystallisation of the residue from hot EtOH afforded 1.04 g of the desired product in 40% yield.
Mp.: 203-205° C.
Mass calc. for C13H14N2O3S: 278.33, found (pos. mode) 279.1, found (neg. mode) 277.1.
According to this general method the following compounds were prepared:
D161: Mp.: 249-250° C.
Mass calc. for C14H16N6O3S: 292.36, found 293.1 (pos. mode), found 291.1 (neg. mode).
D160: Mp.: 188-189° C.
Mass calc. for C14H16N6O3S: 292.36, found 293.1 (pos. mode), found 291.1 (neg. mode).
Typically, a mixture of 90 mg (0.32 mmol) 2-(cyclopropylcarbonyl-amino)-7-oxo-4,5,6,7-tetrahydro-benzo[b]-thiophen-3-carboxylic acid amide and hydroxylamine hydrochloride or an O-substituted hydroxylamine derivative hydrochloride salt (0.71 mmol) in 3 ml EtOH was heated in the microwave at 135° C. for 5 min. The reaction was then poured into water (15 ml) and extracted three times with EtOAc (25 ml). Drying of the combined organic layers over Na2SO4 and evaporation of the solvent yielded after purification by preparative HPLC (Method 1) the isomerically pure solids (Z and E isomer).
According to this general method the following compounds were prepared:
D84: Mp.: 255-256° C.
Mass calc. for C13H15N3O3S: 293.35, found (pos. mode) 294.1.
D104: Mp.: 240-241° C.
Mass calc. for C13H15N3O3S: 293.35, found 294.1 (pos. mode).
D85: Mp.: 213-214° C.
Mass calc. for C14H17N3O3S: 307.37, found (pos. mode) 308.1.
D88: Mp.: 210-211° C.
Mass calc. for C14H17N3O3S: 307.37, found (pos. mode) 308.1.
D72: Mp.: 195-196° C.
Mass calc. for C20H21N3O3S: 383.47, found (pos. mode) 384.2, found (neg. mode) 382.3.
D73: Mp.: 176-177° C.
Mass calc. for C20H21N3O3S: 383.47, found (pos. mode) 384.4, found (neg. mode) 382.3
D93: Mp.: 265-266° C. (dec.).
Mass calc. for C14H17N3O3S: 307.37, found (pos. mode) 308.1.
1 mmol 2-(Cyclopropylcarbonyl-amino)-6-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide was suspended in acetonitrile then 1.5 equivalent K2CO3 and 1.5 equivalent hydroxylamine HCl were added to the mixture and stirred at reflux for 5 h. The solvent was removed and extracted with EtOAc two times. The collected organic layer was washed with water and dried over Na2SO4, and the solvent was evaporated. The crude product was crystallized from isopropanol. If hydrazine is used, then EtOH is the solvent of choice and the reaction is completed after 24 h at room temperature.
According to this general method the following compounds were prepared:
To a solution of 1.86 ml ethyl 2-formyl-1-cyclopropane-carboxylate (1.07 mmol, predominantly trans) in 10 ml EtOH at 0° C. were added 692 mg sodium borohydride (18.3 mmol) in three portions, and the mixture was stirred at room temperature for 20 h. After cooling to 0° C., the reaction was quenched by the addition of 1 M HCl solution (10 ml) and the aqueous layer extracted three times with DCM (20 ml). The combined organic layers were dried over Na2SO4 and after removal of the solvent 1.99 g of a colourless oil was obtained (Yield: 98%).
Mass calc. for C7H12O3: 144.17, found (pos. mode) 145.1
To a suspension of 72 mg sodium hydride (60% in mineral oil, 1.80 mmol) in 1.5 ml abs. THF were slowly added 200 mg ethyl 2-(hydroxymethyl)cyclopropane-carboxylate (1.39 mmol), and the resulting mixture was stirred for 30 min at 0° C. After dropwise addition of RX (methyliodide, allylbromide or benzylchloride, 1.80 mmol), the reaction was stirred for 3 h at 0° C., then for 1 h at r.t and finally quenched by the addition of water (10 ml). The resulting solution was extracted three times with DCM (10 ml) and the combined organic layers were dried over Na2SO4. Evaporation of the solvent and purification by bulb-to-bulb distillation afforded the desired compound as a colourless oil.
Yield: 52%.
Mass calc. for C8H14O3: 158.20, found (pos. mode) 159.2.
NMR (200 MHz, CDCl3): δ 0.85 (m, 1H), 1.23 (m, 1H), 1.26 (t, J=7.3 Hz, 3H), 1.57 (m, 1H), 1.71 (m, 1H), 3.26 (dd, J=10.3, 6.6 Hz, 2H), 3.35 (s, 3H), 3.37 (dd, J=10.3, 6.6 Hz, 1H), 4.12 (q, J=7.3 Hz, 2H).
According to this general method the following compounds were prepared:
Yield: 79%.
Mass calc. for C9H14O3: 184.24.
NMR (200 MHz, CDCl3): δ 0.88 (m, 1H), 1.24 (m, 1H), 1.26 (t, J=7.3 Hz, 3H), 1.58 (m, 1H), 1.71 (m, 1), 3.38 (m, 2H), 3.99 (m, 2H), 4.12 (q, J=7.3 Hz, 2H), 5.24 (m, 2H), 5.90 (m, 1H).
Yield: 43%.
Mass calc. for C14H18O3: 234.30.
NMR (200 MHz, CDCl3): δ.0.86 (m, 1H), 1.22 (m, 1H), 1.26 (t, J=7.3 Hz, 3H), 1.52 (m, 1H), 1.73 (m, 1H), 3.41 (m, 2H), 4.12 (q, J=7.3 Hz, 2H), 4.53 (m, 2H), 7.36 (m, 5H).
To a solution of ethyl cyclopropanecarboxylate (3.01 mmol) in 6 ml THF were added 6 ml 3M NaOH solution and the reaction was heated to reflux for 6 h. After cooling to room temperature, the reaction was acidified with 3M HCl solution to pH 1 and extracted three times with EtOAc (20 ml). Evaporation of the solvent yielded the title compound.
According to this general method the following compounds were prepared:
2-[(Methyloxy)methyl]cyclopropanecarboxylic acid
Yield: 69%
NMR (200 MHz, CDCl3): δ.0.91 (m, 2H), 1.58 (m, 1H), 1.76 (m, 1H), 3.34 (dd, J=10.3, 6.6 Hz, 1H), 3.35 (sb, 3H), 3.45 (dd, J=10.3, 5.9 Hz, 1H).
Yield: quant.
NMR (200 MHz, CDCl3): δ.0.91 (m, 2H), 1.57 (m, 1H), 1.77 (m, 1H), 3.33 (dd, J=10.3, 6.6 Hz, 1H), 3.45 (dd, J=10.3, 5.9 Hz, 1H), 3.99 (d, J=5.9 H, 2H), 5.24 (m, 2H), 5.89 (m, 1H), 10.53 (sbr, 1H).
Yield: 26%
NMR (200 MHz, CDCl3): δ.0.93 (m, 1H), 1.26 (m, 1H), 1.56 (m, 1H), 1.79 (m, 1H), 3.34 (dd, J=10.3, 6.6 Hz, 1H), 3.46 (dd, J=10.3, 5.9 Hz, 1H), 4.51 (s, 2H), 7.30 (m, 5H).
A suspension of 580 mg 2-[(2-allyloxymethyl-cyclopropanecarbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (1.72 mmol), 327 mg p-toluenesulfonic acid (1.72 mmol) and 350 mg palladium on charcoal (10%, 0.33 mmol) in 10 ml methanol and 10 ml water was heated to reflux for 4 h. The reaction mixture was then cooled to room temperature and filtrated through a pad of Celite (solvent EtOAc). The filtrate was washed with brine (30 ml), the organic layer dried over Na2SO4 and the solvent evaporated. After purification by flash column chromatography on silica gel (DCM/MeOH 9:1) 208 mg of the title compound were obtained as a powder (Yield: 41%).
Mass calc. for C13H16N2O4S: 296.35, found (pos. mode) 297.2 found (neg. mode) 295.1.
A solution of 322 mg p-toluenesulfonyl chloride (1.68 mmol) and 208 mg 2-[(2-hydroxymethyl-cyclopropanecarbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide in 7 ml abs. DCM was stirred at room temperature and 272 μl pyridine (2.66 mg, 3.36 mmol) were slowly added. After 19 h stirring at room temperature, water (15ml) was added to the reaction mixture and the resulting solution was extracted three times with EtOAc (15 ml). The combined organic layers were dried over Na2SO4, the solvent evaporated and purification by flash column chromatography on silica gel (DCM/MeOH 96:4) yielded 210 mg of the desired product (Yield: 66%).
Mass calc. for C20H22N2O6S2: 450.54, found (pos. mode) 451.2 found (neg. mode) 449.2.
The product was directly used in the next reaction step without further analysis.
Nucleophilic Substitution
To a solution of 104 mg toluene-4-sulfonic acid 2-(3-carbamoyl-4,7-dihydro-5H-thieno[2,3-c]pyran-2-ylcarbamoyl)-cyclopropylmethyl ester (0.24 mmol) in 2 ml EtOH and 1 ml DCM were added typically, 20-30 eq. of a secondary amine (neat or as commercially available solutions in THF) and the reaction was stirred for 48-72 h at room temperature. After addition of water (10 ml), the resulting solution was extracted three times with EtOAc (15 ml) and the combined organic layer was dried over Na2SO4. Evaporation of the solvent, followed by purification by preparative HPLC (Method 1) afforded the products as off-white solids.
According to this general method the following compounds were prepared:
B197: Mp.: 175-176° C.
Mass calc. for C15H21N3O3S: 323.42, found (pos. mode) 324.2 found (neg. mode) 322.2.
B196: Mp.: 193-194° C.
Mass calc. for C17H23N3O4S: 365.45, found (pos. mode) 366.1 found (neg. mode) 354.2.
B193:
Mass calc. for C18H26N4O3S: 378.50, found (pos. mode) 379.2 found (neg. mode) 377.2.
The ethyl ester (1 mmol) was dissolved in 3 ml abs. THF, then LiNH2 (230 mg, 10 equivalents) was added and the mixture was stirred in a stoppered flask at r.t. for 48 h. The reaction mixture was poured on ice water, and the pH of the solution was adjusted to 5 with 5% HCl. The precipitate was filtered off and washed with cold isopropanol. The reaction can be run also under heterogeneous conditions. Diethyl ether can also be used as solvent.
According to this general method the following compounds were prepared:
53 mg (0.24 mmol) CuBr2 were added to a solution of 60 mg (0.22 mmol) 2-(cyclopropanecarbonyl-amino)-6-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxamide in 2 ml CH3CN. The mixture was stirred at room temperature for 4 h. 10 ml water were added and the mixture extracted three times with 20 ml EtOAc. The combined organic layers were washed with brine, dried over Na2SO4 and the solvent was evaporated. Separation by preparative HPLC (Method 1) afforded the title compound as a light brown solid in 10% yield.
21 mg (0.12 mmol) of N-bromosuccinimide were added to a suspension of 30 mg (0.11 mmol) of 2-(cyclopropanecarbonyl-amino)-6-hydroxy-benzo[b]thiophene-3-carboxamide. The mixture was stirred at room temperature overnight. Water (10 ml) was added and the mixture extracted three times with 15 ml DCM. The combined organic layers were washed with brine, dried over Na2SO4 and the solvent was evaporated. Separation by preparative HPLC (Method 1) afforded the title compound as a yellow solid in 45% yield.
Mp.: 236-240° C. (CH3CN).
Mass calc. for C13H11BrN2O3S: 355.21, found (pos. mode) 355.0 & 357.0, found (neg. mode) 353.0 & 355.0.
When using a 2.2 fold excess of N-bromosuccinimide in acetic acid as a solvent and stirring for 48 h at room temperature, the 5-7-di-bromo-derivative is obtained in 77% yield.
According to this procedure the following compound was prepared:
275 mg (1 mmol) of 2-(2-chloro-acetylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide were treated with 2.5 ml cyclopropylamine and refluxed till the product precipitated. The product was filtered, washed with water, cold isopropanol, diethyl ether and dried (Yield: 61%).
The following compounds were prepared by this method:
To a suspension of 275 mg (1 mmol) of 2-(2-chloro-acetylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide in EtOH, the corresponding thiol (1.3 eq.) and NaOAc (1.3 eq.) wee added and the mixture was stirred at reflux for 2 h. Upon cooling the product precipitated and it was filtered and washed with water and isopropanol (Yield: 75%).
The following compounds were prepared by this method:
To a solution of 1.33 g (5.00 mmol) of 2-(cyclopropanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide and 0.45 g (5.50 mmol) of sodium acetate in 30 ml acetic acid, 0.88 g (5.50 mmol) of bromine was added dropwise at room temperature. After stirring the reaction mixture at room temperature for 1 h the precipitated product was filtered off, washed with 15 ml diisopropyl ether and dried to yield the title compound in 67% yield.
170 mg (0.50 mmol) of 7-bromo-2-(cyclopropanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide, 0.06 g (5.0 mmol) diisopropyl-ethylamine and 5.00 mmol of 2-ethoxy-ethanol were refluxed in 25 ml abs. tetrahydrofuran for 2-4 h. The solvent was evaporated, and the residue was partitioned between ethyl acetate and water (15 ml each). The aqueous phase was extracted two times with 15 ml ethyl acetate, the combined organic phases were washed with 15 ml brine, dried over magnesium sulphate, and the solvent was evaporated. The crude product was crystallized from acetonitrile to give the title compound.
When using N-nucleophiles diisopropyl-ethylamine may be omitted.
According to this general method the following compounds were prepared:
A suspension of 53 mg (1.0 mmol) NH4Cl in 2.5 ml abs. toluene at 0° C. under argon was treated dropwise with 500 μl (1.0 mmol) AlMe3 (2 M solution in toluene). The mixture was stirred at room temperature until the evolution of gas stopped. This solution was added dropwise to a suspension of 100 mg (0.41 mmol) cyclopropanecarboxylic acid (3-cyano-4,5,6,7-tetrahydro-benzo[b]thiophene-2-yl)-amide in 1 ml toluene. The mixture was stirred under reflux for 24 h. After cooling to room temperature the mixture was poured into a slurry of 10 g silica gel in CH2Cl2 and stirred for 10 min. The silica gel was filtered and washed with CH2Cl2. Evaporation of the solvent gave the crude product, which was purified by flash column chromatography on silica gel (eluent DCM/MeOH 10:1 to 4:1) to afford the title product as a colourless solid in 43% yield.
Mp.: >240° C. (decomposition).
Mass calc. for C13H17N3OS: 263.36, found (pos. mode) 264.25, (neg. mode) 262.29.
A solution of 243 mg (0.92 mmol) 2-(cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxamide and 557 mg (1.38 mmol) Lawesson's reagent in 6 ml THF was stirred under reflux for 3 h. The crude mixture was directly purified by flash column chromatography on silica gel (eluent cHex/EtOAc 10:1 to 5:1) to yield the title compound (D99) as yellow crystals in 35%.
Mp.: 182-184° C. (cHex/CH2Cl2),
Mass calc. for C13H16N2OS2: 280.41, found (neg. mode) 279.28; and the title compound (D100) as an orange powder in 18% yield,
Mp.: 169-170° C. (decomposition, cHex/CH2Cl2).
A suspension of 50 mg (0.18 mmol) 2-(cyclopropanecarbonyl-amino)-6-hydroxy-benzo[b]thiophene-3-carboxamide and 35 μl (0.20 mmol) diisopropylethylamine in 2.5 ml THF was treated with 14 μl (0.20 mmol) acetyl chloride and stirred at room temperature overnight. 10 ml 0.5 M HCl solution was added and the mixture extracted three times with 20 ml EtOAc. The combined organic layers were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by flash column chromatography on silica gel (eluent cHex/EtOAc 1:1) to afford a colourless solid in 42% yield.
Mp.: 223-224° C. (ethanol).
Mass calc. for C15H14N2O4S: 318.35, found (pos. mode) 319.2, found (neg. mode) 317.2.
To a solution of 70 mg (0.19 mmol) diethyl-carbamic acid 3-carbamoyl-2-(cyclopropanecarbonyl-amino)-benzo[b]thiophen-6-yl ester and 113 μl (0.75 mmol) N,N,N′,N′-tetramethylethylenediamine in 2 ml THF at −70° C. were added 533 μl (0.75 mmol) s-BuLi (1.4 M solution in cHex) dropwise. The mixture was stirred at −70° C. for 45 min, then slowly heated to room temperature over 2 h and stirred at room temperature for another 2 h. The mixture was treated with 10 ml sat. NH4Cl solution and extracted three times with 30 ml EtOAc. The combined organic layers were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by preparative HPLC (Method 1) to afford the title product as a colourless solid in 15% yield.
Mp.: 233-235° C. (H2O/CH3CN).
Mass calc. for C18H21N3O4S: 375.45, found (pos. mode) 376.2, found (neg. mode) 374.3.
In the procedure described for 2-(cyclopropanecarbonyl-amino)-6-hydroxy-benzo[b]thiophene-3,7-dicarboxylic acid 3-amide 7-diethylamide, the title compound was isolated as a second product by preparative HPLC (Method 1) to afford the title product as a colourless solid in 11% yield.
Mp.: 227-228° C. (H2O/CH3CN).
Mass calc. for C18H21N3O4S: 375.45, found (pos. mode) 376.2, found (neg. mode) 374.2.
To a solution of 100 mg (0.25 mmol) 2-(cyclopropanecarbonyl-amino)-6-[2-(tetrahydropyran-2-yloxy)-ethoxy]-benzo[b]thiophene-3-carboxamide in 5 ml methanol was added 180 mg Amberlyst 15 (H+-form). The mixture was stirred at 45° C. for 2 h, the Amberlyst 15 filtered off and the solvent evaporated. Recrystallization of the residue from ethanol/methanol (2:1) afforded the product as a colourless solid in 91% yield.
Mp.: 203-205° C. (MeOH/EtOH).
Mass calc. for C15H16N2O4S: 320.37, found (pos. mode) 321.2.
Compound A49 was prepared according to this method (Yield: 50%).
Mp.: 250-254° C.
NMR, δ (ppm): 11.88 (s, 1H), 7.93 (s, 1H), 7.86 (s, 2H), 4.88 (t, J=5.58 Hz, 1H), 4.05 (t, J=5.13 Hz, 2H), 3.78 (m, 2H), 2.07 (m, 1H), 0.96 (m, 4H).
50 mg (0.18 mmol) 2-(cyclopropanecarbonyl-amino)-6-oxo-4,5,6,7-tetrahydro-benzo[b]thio-phene-3-carboxamide was suspended in 2 ml CH2Cl2, and 24 μl (0.18 mmol) diethylamino sulfur trifluoride was added dropwise at 0° C. The mixture was stirred at room temperature overnight. 10 ml sat. NH4Cl-solution was added and the mixture extracted three times with 20 ml EtOAc. The combined organic extracts were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by flash column chromatography on silica gel (eluent cHex/EtOAc 1:3). The title compound was isolated as a colourless solid in 12% yield.
Mass calc. for C13H14F2N2O2S: 300.33, found (pos. mode) 301.19, found (neg. mode) 299.20.
Using the same procedure described for compound D128 the following compound was prepared:
D66: Mp.: 207-208° C. (H2O/CH3CN).
Mass calc. for C13H15FN2O2S: 282.34, found (pos. mode) 283.1, found (neg. mode) 281.1.
40 μl (0.60 mmol) CISO3H was added dropwise to a suspension of 50 mg (0.23 mmol) cyclopropanecarboxylic acid (4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-amide in 1 ml CH2Cl2 at 0° C. The resulting solution was stirred at 0° C. for 1.5 h, then 0.5 ml of water were added. Slow evaporation of the solvents under inert atmosphere afforded the title compound as a colourless solid in 85% yield.
Mp.: 225-230° C. (methanol, decomposition).
Mass calc. for C12H15NO4S2: 301.39, found (pos. mode) 302.2, found (neg. mode) 300.2.
A suspension of 32 mg (0.11 mmol) 2-(cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-sulfonic acid and 2 drops of DMF in 2 ml THF was treated dropwise with 10 μl (0.12 mmol) oxalyl chloride and stirred for 1.5 h at room temperature. 1 ml (0.5 mmol) ammonia (0.5 M solution in dioxane) was added and the mixture stirred at room temperature for 24 h. The solvent was evaporated and the crude product purified by flash column chromatography on silica gel (eluent cHex/EtOAc 4:1) to afford the title compound as a colourless solid in 25% yield.
Mp.: 235-238° C. (decomposition).
Mass calc. for C12H16N2O3S2: 300.40, found (pos. mode) 301.2, found (neg. mode) 299.2.
To a suspension of 13 mg (0.31 mmol) NaH (60% dispersion in mineral oil) in 2 ml DMF was added 11 μl (0.14 mmol) piperidine. The mixture was stirred at room temperature for 30 min, cooled to 0° C. and treated with a solution of 35 mg (0.10 mmol) (6-R,S)-2-amino-3-carbamoyl-4,5,6,7-tetrahydro-benzo[b]thiophene-6-carboxylic acid ethyl ester in 2 ml DMF. The mixture was stirred at room temperature for 4 days. 15 ml 0.5 M HCl solution were added and the mixture extracted four times with 40 ml EtOAc. The combined organic extracts were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the product as a colourless solid in 90% yield.
Mp.: >230° C. (EtOAc, decomposition).
Mass calc. for C14H16N2O4S: 308.36, found (pos. mode) 309.1.
A solution of 50 mg (0.15 mmol) (6-R,S)-2-amino-3-carbamoyl-4,5,6,7-tetrahydro-benzo[b]thiophene-6-carboxylic acid ethyl ester in 1.5 ml THF was treated with 1 ml (1.00 mmol) diisobutylaluminium hydride (1 M solution in DCM) at 0° C. The mixture was stirred at 0° C. for 30 min and then allowed to stir at room temperature for 4 h. The mixture was treated with 10 ml sat. NH4Cl solution and extracted three times with 30 ml EtOAc. The combined organic extracts were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by preparative HPLC (Method 1) to afford the product as a colourless solid in 34% yield.
Mp.: 215-222° C. (CH3CN/H2O, decomposition).
Mass calc. for C14H18N2O3S: 294.38, found (pos. mode) 295.1.
A suspension of 47 mg (0.14 mmol) (6-R,S)-2-amino-3-carbamoyl-4,5,6,7-tetrahydro-benzo[b]thiophene-6-carboxylic acid ethyl ester in 2 ml THF was treated with 233 μl (0.70 mmol) MeMgCl (3 M solution in THF) at 0° C. The mixture was stirred at room temperature for 6 h. The mixture was treated with 10 ml sat. NH4Cl solution and extracted three times with 30 ml EtOAc. The combined organic extracts were washed with brine and dried over Na2SO4. Evaporation of the solvent gave the crude product, which was purified by preparative HPLC (Method 1) to afford the product as a colourless solid in 51% yield.
Mp.: 196-197° C. (CH3CN/H2O).
Mass calc. for C16H22N2O3S: 322.43, found (pos. mode) 323.1.
480 mg 2-(cyclopropylcarbonyl-amino)-7-oxo-4,5,6,7-tetrahydro-benzo[b]thiophen-3-carboxylic acid amide (1.73 mmol) and 195 μl bromine (3.79 mmol) in 15 ml chloroform were heated to reflux for 2.5 h. The reaction was then poured into an aqueous sodium thiosulfate solution (20 ml) and extracted three times with EtOAc (20 ml). Drying of the combined organic layers over Na2SO4 and evaporation of the solvent gave a brown solid, which was further purified by column chromatography on silica gel (eluent DCM/MeOH 95:5). 585 mg of a brown solid were obtained in 78% yield.
Mp.: 196-197° C. (dec.).
Mass calc. for C13H12Br2N2O3S: 436.12, found (pos. mode) 434.9, 436.9 & 438.8, found (neg. mode) 432.9, 434.9 & 436.9.
To a mixture of 20 mg 6,6-dibromo-2-(cyclopropylcarbonyl-amino)-7-oxo-4,5,6,7-tetrahydro-benzo[b]thiophen-3-carboxylic acid amide (45.9 μmol) in 1.0 ml dioxane were added 20 mg potassium carbonate (145.0 μmol) dissolved in 600 μl water, and the resulting yellow solution was heated in the microwave at 110° C. for 30 min. The reaction was poured into water (10 ml) and extracted three times with n-butanol (10 ml). The combined organic layers were evaporated to dryness and the resulting brown oil purified by preparative HPLC (Method 1). 6.0 mg of the desired compound were isolated (Yield: 37%).
Mp.: 235-236° C.
Mass calc. for C13H11BrN2O3S: 355.21, found (pos. mode) 355.1 & 357.1, found (neg. mode) 353.1 & 355.1.
3.8 g (17.1 mmol) of methyl 3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropane carboxylate were dissolved in 20 ml of dichloromethane. 10 ml of trifluoroacetic acid (TFA) were slowly added via syringe at room temperature, and the mixture was stirred for 2 h. After addition of 20 ml of 3N NaOH, keeping the pH below 2, 50 ml of water were added, and the mixture was extracted three times with dichloromethane. The organic layer was dried over Na2SO4. Filtration and evaporation of the solvent gave 3.28 g (92%) of a golden liquid, that was used without further purification.
301.6 mg (2.7 mmol) of 2,2-dimethylcyclopropane carboxamide were refluxed for 24 h in 50 ml of 10% aqueous KOH, then stirred at room temperature for another 24 h. The mixture was acidified with concentrated HCl to a pH of 1, then extracted three times with EtOAc. The organic layer was dried over Na2SO4. Filtration and evaporation of the solvent gave a pale yellow liquid, that was used without further purification.
Mass calc. for C6H10O2: 114.07, found (pos. mode) 115.27.
300 mg (1.25 mmol) of tert-butyl 2,2-dichloro-3,3-dimethylcyclopropanecarboxylate were dissolved in 5 ml of dichloromethane. 5 ml of trifluoroacetic acid (TFA) were slowly added via syringe at room temperature, and the mixture was stirred for 1 h. After addition of 3N NaOH, keeping the pH below 2, 20 ml of water were added, and the mixture was extracted three times with dichloromethane. The organic layer was dried over Na2SO4. Filtration and evaporation of the solvent gave 215 mg (94%) of a brown oil, that was used without further purification.
NMR (200 MHz, DMSO-d6): δ 1.35 (s, 6H), 2.48 (s, 1H).
5 g (44 mmol of monomer) of the dimer of 2-hydroxycyclohexanone, 4.8 ml (44 mmol) of benzylamine, and a spatula tip of p-toluenesulfonic acid were dissolved in a mixture of 50 ml of trimethylorthoformate and 50 ml of abs. THF, and heated under reflux for 4 h. After cooling, 2.2 ml of piperidine were added to the solution, followed by dropwise addition of 3.7 g (44 mmol) of cyanoacetamide in 100 ml of abs. methanol. The solution was heated under reflux for 4 h, and subsequently allowed to cool over night. 100 ml of water were added, and the mixture was extracted three times with EtOAc. The organic layer was dried over Na2SO4. Filtration and evaporation of the solvents gave a crude material that was recrystallized from aqueous ethanol, giving 591 mg (5%) of a pale white solid.
Mp.: 164-165° C. (aq. EtOH)
Mass calc. for C16H19N3O: 269.35, found (pos. mode) 270.44.
To a solution of 211.4 mg (0.79 mmol) of 2-amino-1-benzyl-4,5,6,7-tetrahydro-1H-indole-3-carboxamide and 300 μl (1.73 mmol; 2.2 equivalents) of diisopropylethylamine (DIPEA) in 20 ml of abs. THF under nitrogen atmosphere were added 85 μl (0.94 mmol; 1.2 equivalents) of cyclopropanecarbonyl chloride via syringe at room temperature. The mixture was stirred over night. The solution was diluted with 20 ml of water and extracted several times with EtOAc. The organic layer was dried over Na2SO4. Filtration and evaporation of the solvent gave crude material that was recrystallized from chloroform, giving 17.3 mg (6.5%) of a white solid.
Mp.: >250° C. (decomp.)
Mass calc. for C20H23N3O2: 337.42, found (neg. mode) 336.40.
In a distillation setup, 98.1 mg (0.29 mmol) of ethyl 2-(2-furoylamino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylate and 2.5 ml of ethanolamine were heated at 100° C. for 4 h. The developing ethanol was continuously distilled from the mixture, additionally applying a slow stream of nitrogen gas. After cooling, the excess of ethanolamine was removed in a Kugelrohr distillation apparatus. The crude residue was recrystallized from ethanol to give 70.2 mg (72%) of fine yellow needles.
Mp.: 194-195° C. (EtOH)
Mass calc. for C16H18N2O4S: 334.40, found (neg. mode) 333.28.
To a solution of 318 mg of 2-(3-Pyridin-4-yl-ureido)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (1 mmol) in 1 ml acetonitrile, 142 mg of iodomethane (1 mmol) were added. The mixture was heated for 15 minutes at 100° C. in a closed reaction vessel. The precipitate was filtered off, washed with diethyl ether and dried.
HPLC method B; retention time: 2.05; M−: 331.21
The following compound was also prepared:
1-Benzyl-4-[3-(3-carbamoyl-4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-ureido]-pyridinium Bromide (B72): HPLC method B; retention time: 2.6; M+: 410.16
Synthesis of 2-(Cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]-thiophene-3-carboxylic acid methylamide (D185)
293 mg (1.00 mmol) of 2-(cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid ethyl ester was stirred in 2 ml of a solution containing 33% of methylamine in ethanol at 0° C. for 48 h. The reaction mixture was evaporated to dryness and the product was purified by column chromatography on silica gel (Yield: 22%).
The following compounds were prepared by this method:
To a mixture of 150 mg (0.53 mmol) of N-(3-carbamoyl 4,7-dihydro-5H-thieno[2,3c]pyran-2-yl)oxalamic acid methylester in 5 ml methanol, 1 ml of conc. ammonium hydroxide was added, and the reaction mixture was stirred overnight in a stoppered flask. The solvent was evaporated and the residue was triturated with 0.5 mL methanol and filtered to obtain the title compound in 96% yield.
HPLC method B; retention time: 2.19; M−: 268.11
The above outlined general method 19 is exemplified by the synthesis of A67.
D235 also named as phenyl 7′-(aminocarbonyl)spiro[1,3-dioxolane-2,3′-[9]thiabicyclo[4.3.1]deca[6(10),7]dien]-8′-ylcarbamate was prepared according to procedure 19.1C. After stirring the reaction mixture, the reaction mixture was diluted with water (60 ml), and the mixture was extracted with EtOAc (3×30 ml). The combined organic layers were washed with brine and dried over Na2SO4. Evaporation of the solvent in vacuum at 20° C., the residue was treated with diethyl ether, the precipitate was filtered off and washed with diethyl ether affording 3.53 g (94%) of the title compound.
HPLC method B; retention time: 3.57; M−: 373.42
19.1B Preparation of (3-carbamoyl-6,6-ethylenedioxy-4,5,6,7tetrahydro-benzo[b]thiophen-2-yl)-carbamic acid ethyl ester
This compound was prepared according to 19.1A or 19.1C using dry pyridine as solvent (Yield: 0.27=85%)
NMR, δ (ppm): 10.92 (s, 1H), 7.20 (broad s, 2H), 4.15 (m, 2H), 3.93 (s, 4H), 2.82 (m, 4H), 1.82 (m, 2H), 1.23 (t, J=7.08 Hz, 3H).
To a solution of 2-amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (4.45 g, 22.44 mmol) in 90 ml of tetrahydrofuran, pyridine (2.3 ml, 28.4 mmol) was added and stirred at 5° C. To the reaction mixture phenyl chloroformate (3.4 ml 24.58 mmol) was added dropwise at 5° C. After stirring for 3 h the mixture was diluted with water (50 ml), the solid was collected, washed with water, and dried in vacuum, affording 5.76 g (yield: 81%) of the title compound as a white solid.
A suspension of 3.07 g (8.2 mmol) (3-carbamoyl-6,6-ethylenedioxy-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-carbamic acid phenyl ester in 65 ml CH3CN was treated with 2.93 g (17.1 mmol) CuCl2.2 H2O at room temperature. The mixture turned brown immediately and was stirred at room temperature for 1 h. The solvent was evaporated in vacuum at 20° C. and the residue taken up in 100 ml EtOAc and extracted with 6×100 ml 0.5 M HCl solution. The organic layer was washed with brine and dried over Na2SO4. Evaporation of the solvent, the residue was treated with diethyl ether, the precipitate was filtered off and washed with diethyl ether affording 2.16 g (80.5%) of the title compound.
HPLC method B; retention time: 3.38; M−: 327.35
According to this Method, (3-carbamoyl-6-hydroxy-benzo[b]thiophen-2-yl)-carbamic acid ethyl ester was Prepared.
Yield: 0.18 (65%)
NMR, δ (ppm): 11.24 (s, 1H), 9.51 (s, 1H), 7.72 (d, J=8.79 Hz, 1H), 7.24 (broad s, 2H), 7.23 (d, J=2.01 Hz, 1H), 6.87 (dd, J1=2.13 Hz, J2=8.76 Hz, 1H ), 4.21 (m, 2H), 1.27 (t, J=7.08 Hz, 3H).
To a mixture of (3-carbamoyl-4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-carbamic acid phenyl ester (95.5 mg, 0.30 mmol) and 2 ml of THF, cyclopentylamine (0.75 mmol) was added. After stirring at ambient temperature until the reaction was completed (2-6 h), the mixture was poured into 1M NaOH, extracted twice with EtOAc, and the combined organic layers were dried over Na2SO4, filtered and concentrated. Purification of the residue by chromatogrqnhy (1 mm plate, eluent:chloroform-EtOAc 3:2), followed by concentration and trituration of the residue with ether afforded 2-(3-cyclopentyl-ureido)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide as a colourless solid.
Yield: 81%
According to this procedure the following compounds were prepared:
The following compounds were prepared by this method using 25 mg 4-dimethylaminopyridine as catalyst:
To a mixture of (3-carbamoyl-6-hydroxy-benzo[b]thiophen-2-yl)-carbamic acid phenyl ester (131 mg, 0.40 mmol), 0.2 ml DMSO and 2.5 ml of THF, 2-thiopheneethylamine (0.117 ml 1.0 mmol) was added. After stirring at ambient temperature until the reaction was completed (24-48 h), the mixture was poured into 10 ml of 1N HCl solution and extracted twice with EtOAc (20 ml). The combined organic layers were dried over Na2SO4, filtered and concentrated. Purification of the residue by chromatography (1 mm plate, eluent: chloroform-methanol 9:1), followed by concentration and trituration of the residue with ether afforded the title compound as a white solid.
Yield: 39%.
NMR (300 MHz, DMSO-d6 ppm): 10.77 (s, 1H), 9.34 (s, 1H), 7.96 (s, 1H), 7.66 (d, 1H), 7.41 (s, 2H), 7.34 (d, 1H), 7.14 (s, 1H), 6.96 (m, 2H), 6.82 (d, 1H), 3.38 (t, 2H), 2.98 (t, 2H)
According to this general method the following compounds were prepared:
The following compounds were prepared by this method using (3-carbamoyl-6-oxo-ethylenylketal-4,5,6,7-tetrahydro-benzo[b]thiophen-2-yl)-carbamic acid phenyl ester (m.p. 175° C.) as starting material:
To a solution of the corresponding ketal derivative (1 mmol) in 10 ml acetonitrile 1.7 g (10 mM) fine powdered CuCl2 was added with stirring. The reaction mixture was heated to 80° C. and stirred for 1 hour. Then the solvent was evaporated 10 ml 1N hydrochloric acid was added to the residue and extracted with 3×10 ml ethyl acetate.
Organic phase was dried over sodium sulfate and evaporated to dryness. The resulted dark crystalline material was washed with 0.5 ml cold ethylacetate and the formed crystals were isolated. The crude products may be recrystallized from acetonitrile if necessary.
According to this general method the following compounds were prepared:
To a solution of 196 mg (1 mmol) of 2-amino-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide in 5 ml abs. dioxane, 120 mg (1.2 mmol) of succinic anhydride were added. The mixture was stirred at 100° C. for 10 h. The reaction mixture was poured into cold water, the resulting precipitate was collected, washed with water and dried (Yield: 70%).
HPLC method B; retention time: 2.94; M−: 295.17
The following compounds were prepared by this method:
HPLC method B; retention time: 2.17; M−: 297.08
HPLC method B; retention time: 2.17; M−: 295.06
HPLC method B; retention time: 2.89; M−: 293.08
To a solution of 240 mg of 2-{[(2E)-3-phenylprop-2-enoyl]amino}-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxamide in 10 ml of ethanol, 50 mg of 10% palladium/charcoal were added and the mixture hydrogenated at room temperature and normal pressure. The catalyst was removed by filtration over Celite and the solvent was evaporated (Yield: 93%).
HPLC method A; retention time: 1.86; M+: 331.13
1 mM tetrahydro-pyran-4-one, 106 μl (1 mM) ethyl-cyano-acetate, 0.15 mM ammonium-acetate, and 0.2 mM acetic acid where dissolved in 3 ml benzene and stirred at reflux temperature in a round-bottomed flask equipped with water-remover trap, for 3 hours. The reaction mixture was washed with 2 ml 10% K2CO3 solution, dried, and evaporated to dryness. The solid material was dissolved in 1.5 ml EtOH and was stirred with 1.05 mM sulphur and 0.575 mM morpholine at 45-50° C., for 4 hours. The reaction mixture was evaporated to dryness, washed with n-hexane and isopropylalcohol. This reaction step was developed starting from a procedure described by Gewald, K; Schinke, E; Böttcher, H; Chem. Ber. 1966, 99, 974.
Yield: 57%
NMR: 7.28 (s, 2H), 4.43 (s, 2H), 4.16 (q, 2H), 3.79 (t, 2H), 3.67 (t, 2H), 1.25 (t, 3H)
1 mM cyclopropanecarbonyl chloride was added dropwise to a well stirred, 15 ml ethylacetate solution of 301 mg (1.00 mM) 2-amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester. The reaction mixture was stirred for 3 hours, then diluted to 50 ml, washed two times with water, dried with MgSO4, and evaporated to dryness. The product was washed with n-hexane and isopropanol.
Yield: 42%
NMR: 11.19 (s, 1H), 4.60 (s, 2H), 4.30 (q, 2H), 3.84 (t, 2H), 2.78 (t, 2H), 2.03 (m, 1H), 1.33 (t, 3H), 0.93 (m, 4H)
Analogous to this method the following compounds were also synthesized:
2-[(Furan-2-carbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 57%), NMR: 11.90 (s, 1H), 8.06 (s, 1H), 7.39 (d, 1H), 6.79 (dd, 1H), 4.66 (s, 2H), 4.35 (q, 2H), 3.86 (t, 2H), 2.82 (t, 2H), 1.35 (t, 3H);
2-[(Adamantane-1-carbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 67%), NMR: 11.36 (s, 1H), 4.62 (s, 2H), 4.32 (q, 2H), 3.84 (t, 2H), 2.79 (t, 2H), 2.06 (bs, 2H), 1.90 (s, 8H), 1.72 (s, 6H), 1.33 (t, 3H);
2-(Cyclohexanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 70%), NMR: 11.10 (s, 1H), 4.61 (s, 2H), 4.30 (q, 2H), 3.84 (t, 2H), 2.78 (t, 2H), 1.90 (d, 2H), 1.69 (m, 3H), 1.43-1.18-(m, 9H);
2-[(2-Methyl-cyclopropanecarbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 57%), NMR: 11.15 (s, 1H), 4.60 (s, 2H), 4.25 (q, 2H), 3.64 (t, 2H), 2.78 (t, 2H), 1.80 (m, 1H), 1.33 (t, 3H), 1.11 (d, 3H), 0.79 (m, 1H);
2-(Cyclobutanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 76%), NMR: 10.91 (s, 1H), 4.61 (s, 2H), 4.28 (q, 2H), 3.83 (t, 2H), 3.44 (m, 1H), 2.78 (bs, 2H), 2.23 (m, 4H), 1.97 (m, 1H), 1.83 (m, 1H), 1.31 (t, 3H);
2-Acetylamino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 85%), NMR: 10.93 (s, 1H), 4.61 (s, 2H), 4.29 (q, 2H), 3.84 (t, 2H), 2.77 (t, 2H), 2.24 (s, 3H), 1.32 (t, 3H);
2-But-2-enoylamino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 76%), NMR: 11.02 (s, 1H), 6.90 (m, 1H), 6.35 (dd, 1H), 4.63 (s, 2H), 4.30 (q, 2H), 3.84 (t, 2H), 2.79 (t, 2H), 1.92 (s, 3H), 1.89 (s, 3H), 1.32 (t, 3H);
2-(2-Methyl-butyrylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 69%), 11.05(s, 1H),4.62(s,2H),4.30(q,2H), 3.84(t,2H), 2.80(t,2H), 2.59 (m,1H), 1.64(m, 1H), 1.50(m, 1H), 1.32(t,3H), 1.14(d,3H), 0.87(t,3H);
2-(2,2-Dimethyl-propionylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (yield 76%), NMR: 11.05(s, 1H),4.62(s,2H),4.30 (q,2H), 3.84(t,2H), 2.80(t,2H), 2.59(m, 1H), 1.64(m,1H), 1.50(m, 1H), 1.32(t,3H), 1.14(d,3H), 0.87 (t,3H);
2-(2-Chloro-acetylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 82%), NMR: 11.64 (s, 1H), 4.64 (s, 2H), 4.61 (s, 2H), 4.32 (q, 2H), 3.85 (t, 2H), 2.80 (t, 2H), 1.32 (t, 3H);
2-(3,4-Difluoro-benzoylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 79%), NMR: 11.89 (s, 1H), 7.95 (m, 1H), 7.76 (m, 2H), 4.67 (s, 2H), 4.34 (q, 2H), 3.87 (t, 2H), 2.82 (t, 2H), 1.34 (t, 3H);
2-Isobutyrylamino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 80%), NMR: 11.08 (s, 1H), 4.62 (s, 2H), 4.30 (q, 2H), 3.84 (t, 2H), 2.78 (m, 3H), 1.32 (t, 3H), 1.17 (d, 6H);
2-(Cyclopentanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester (Yield 77%), NMR: 11.04 (s, 1H), 4.61 (s, 2H), 4.29 (q, 2H), 3.84 (t, 2H), 2.99 (m, 1H), 2.78 (t, 2H), 1.91 (m, 2H), 1.65 (m, 6H), 1.31 (t, 3H).
1 mmol 2-Amino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester was dissolved in 10 ml benzene and 348 μl (2.5 equivalent) triethylamine, 195 μl (2.5 equiv.) methanesulfonyl chloride was added. The reaction mixture was refluxed for 8 hours. The mixture was extracted with 1×15 ml water, 1×15 ml NaHCO3, then 1×15 ml water, 1×15 ml 1N HCl and saturated NaCl solution. The organic layer was dried above MgSO4, the solvent was evaporated to vacuo and the residue was crystallized from hexane-isopropanol. (TLC-Eluent:Hexan-Ethylacetate: 2:1)
Yield: 65%, NMR: 11.03 (s, 1H), 4.73 (s, 2H), 4.28 (q, 2H), 3.89 (t, 2H), 3.53 (s, 3H), 2.83 (t, 2H), 1.29 (t, 3H).
269 mg (1 mmol) 2-Acetylamino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester was dissolved in 15 ml acetic acid and 82 mg (1 mmol) sodium-acetate was added to the mixture, then heated to 55° C. 159 mg bromine in 15 ml acetic acid was added slowly to the mixture. After one hour stirring it was evaporated under reduced pressure and extracted three times with ethyl acetate and 15 ml water. The organic layer was washed with 10 ml NaHCO3 solution and dried with MgSO4. The solution was evaporated under reduced pressure and the product was crystallized from hexane. The product was washed with IPA, and recrystallized with diisopropyl-ether. Yield: 39% NMR: 10.92 (s, 1H), 4.83 (d, 1H), 4.73 (d, 1H), 4.49 (d, 1H), 4.29 (q, 2H), 3.90 (d, 1H), 3.65 (d, 1H), 2.24 (s, 3H), 1.32 (t, 3H).
The compound 2-(cyclopropanecarbonyl-amino)-7-hydroxy-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester was synthesized in a analogous reaction. Yield: 62%, NMR: 11.20 (s, 1H), 5.65 (s, 1H), 4.93-4.65 (m, 2H), 4.34 (q, 2H), 4.24-4.03 (m, 3H), 2.10 (m, 1H), 1.38 (t, 3H), 0.93 (m, 4H).
470 mg (12.00 mM) sodium amide was added to the solution of 293 mg (1.00 mM) 2-(cyclopropanecarbonyl-amino)-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid ethyl ester in 8 ml abs. tetrahydrofurane. The air-tightly closed reaction mixture was stirred at room temperature for 72 hours.
After the starting material disappeared, the pH of the reaction mixture was set to 5-6 with ice cold, 1 N HCl, the precipitated product was filtered off, washed twice with 5 ml n-hexane and dried.
Yield: 89% white, or off-white crystals; NMR: 11.75 (s, 1H), 4.62 (s, 2H), 3.83 (t, 2h), 2.79 (t, 2H), 1.89 (m, 1H), 0.87 (m, 4H)
1 mmol 2-(Cyclopentanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid ethyl ester was dissolved in 3 ml abs. THF, then 230 mg (10 equivalent) LiNH2 was added and the mixture was stirred in a stoppered flask at r.t. for 48 hours. The reaction mixture was poured on ice water, the pH of the solution was adjusted to 5 with 5% HCl. The precipitated crystals were filtered out and washed with cold isopropanol. (TLC Eluent:chloroform-MeOH 10:1)
Yield: 79%, NMR: 11.73 (s, 1H), 7.2 (bd, 2H), 4.63 (s, 2H), 3.83 (t, 2H), 2.88 (m, 1H), 2.80 (t, 2H), 1.89 (m, 2H), 1.64 (m, 6H).
The following compounds were also prepared by this method:
2-(2-Methyl-butyrylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B14), Yield: 67%, NMR: 11.77 (s, 1H), 7.2 (bs, 2H), 4.63 (s, 2H), 3.83 (t, 2H), 2.80 (t, 2H), 2.46 (m, 1H), 1.60 (m, 1H), 1.47 (m, 1H), 1.12 (d, 3H), 0.85 (t, 3H);
2-(Cyclobutanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B15), Yield: 74%, NMR: 11.63 (s, 1H), 7.2 (bs, 2H), 4.63 (s, 2H), 3.83 (t, 2H), 3.33 (m, 1H), 2.81 (t, 2H), 2.20 (m, 4H), 1.97 (m, 1H), 1.83 (m, 1H);
2-[(2-Phenyl-cyclopropanecarbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B3), Yield: 73%, NMR: 11.26 (s, 1H), 7.32-7.19 (m, 5H), 4.62 (s, 2H), 4.28 (q, 2H), 3.84 (t, 2H), 2.78 (t, 2H), 1.55 (m, 1H), 1.43 (m, 1H), 1.30 (t, 3H);
2-But-2-enoylamino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B16), Yield: 49%, NMR: 11.64 (s, 1H), 7.3 (bd, 2H), 6.83 (m, 1H), 6.23 (dd, 1H), 4.64 (s, 2H), 3.83 (t, 2H), 2.80 (t, 2H), 1.89 (d, 3H);
2-(3-Methyl-but-2-enoylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B17), Yield 31%, NMR: 11.56 (s, 1H), 7.2 (bs, 2H), 5.93 (s, 1H), 4.64 (s, 2H), 3.83 (t, 2H), 2.80 (t, 2H), 2.16 (s, 3H), 1.90 (s, 3H);
2-(2,2-Dimethyl-propionylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B18), Yield 32%, NMR: 12.33 (s, 1H), 7.2 (bs, 2H), 4.64 (s, 2H), 3.83 (t, 2H), 2.83 (t, 2H), 1.22 (s, 9H);
2-(3,4-Difluoro-benzoylamino)-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B19), Yield: 70%; NMR: 13.01 (s, 1H), 7.88 (m, 1H), 7.70 (m, 2H), 7.30 (bs, 2H), 4.69 (s, 2H), 3.86 (t, 2H), 2.86 (t, 2H);
2-Isobutyrylamino-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B2), Yield: 61%, NMR: 11.81 (s, 1H), 7.2 (bd, 2H), 4.63 (s, 2H), 3.83 (t, 2H), 2.81 (t, 2H), 2.67 (m, 1H), 1.14 (d, 6H);
2-[(2-Methyl-cyclopropanecarbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B4), Yield: 53%, NMR: 11.71 (s, 1H), 7.5 (bs, 1H), 7.0 (bs, 1H), 4.61 (s, 2H), 3.82 (t, 2H), 2.79 (t, 2H), 1.64 (m, 1H), 1.09 (d, 3H), 0.73 (m, 1H);
2-[(Furan-2-carbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B5), Yield: 34%, NMR: 12.72 (s, 1H), 8.02 (d, 1H), 7.7 (bs, 1H), 7.31 (d, 1H), 7.2 (bs, 1H), 6.76 (dd, 1H), 4.67 (s, 2H), 3.85 (t, 2H), 2.86 (t, 2H);
2-[(Adamantane-1-carbonyl)-amino]-4,7-dihydro-5H-thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B6), Yield: 61%, NMR: 12.23 (s, 1H), 7.3 (b, 2H0, 4.63 (s, 2H), 3.83 (t, 2H), 2.83 (t, 2H), 2.03 (s, 3H), 1.86 (s, 5H), 1.70 (s, 5H);
2-(Cyclohexanecarbonyl-amino)-4,7-dihydro-5H -thieno[2,3-c]pyran-3-carboxylic acid amide (Compound B7), Yield: 63%, NMR: 11.79 (s, 1H), 7.2 (bd, 2H), 4.63 (s, 2H), 3.82 (t, 2H), 2.80 (t, 2H), 2.41 (m, 1H), 1.89-1.62 (m, 5H), 1.40-1.17 (m, 5H).
Sulfurylchloride (13 mmol) was added dropwise to DMF (13 mmol) at 0° C. under Argon. The mixture was stirred for 30 min at 0° C. and 2-(cyclopropanecarbonyl-amino)-4,7-dihydro-5H-thieno[2,3-c]pyrane (10 mmol) in 2 ml DCM added. The mixture was stirred for 1 h at r.t., diluted with 2 ml of THF and treated with an excess of NH3 (2 M solution in dioxane, 10 ml, 20 mmol). The mixture was stirred at room temperature overnight. Evaporation of the solvent and recrystallization afforded the title compound.
2-Amino-3-cyano-4,7-dihydro-5H-furo[2,3-c]pyrane (0.66 mmol) and cyclopropylcarbonyl chloride (0.8 mmol) were dissolved in 10 mL of abs. THF. Diisopropylethylamine (0.8 mmol) was added via syringe, and the mixture was stirred overnight at room temperature. After dilution with 20 ml of water, the aqueous phase was extracted four times with ethylacetate, the organic layer washed once with water, dried over sodium sulfate and the solvents evaporated. Recrystallization of the crude material from hot ethanol gave the desired product.
1-Benzyl-2-amino-3-cyano-4,7-dihydro-5H-pyrrolo[2,3-c]pyrane (4.1 mmol) and cyclopropylcarbonyl chloride (4.9 mmol) were dissolved in 10 ml of abs. THF. 1.5 ml of diisopropylethylamine were added via syringe, and the mixture was stirred overnight at room temperature. After dilution with 20 ml of water, the aqueous phase was extracted four times with ethylacetate, the organic layer washed once with water, dried over sodium sulfate and the solvents evaporated. Recrystallization of the crude material from hot ethanol gave the desired product.
2-(Cyclopropanecarbonyl-amino)-3-cyano-4,7-dihydro-5H-furo[2,3-c]pyrane (4.3 mmol), 1 ml of water, and 7 ml of boron trifluoride-acetic acid complex are heated at 120° C. for 10 minutes. After cooling, the reaction mixture is treated with 50 ml of 6 N sodium hydroxide solution, the aqueous mixture is extracted with ethylacetate, dried over sodium sulfate and the solvents evaporated. The crude material can be recrystallized from hot ethanol.
1-Benzyl-2-(cyclopropanecarbonyl-amino)-3-cyano-4,7-dihydro-5H-pyrrolo[2,3-c]pyrane (4 mmol), 1 ml of water, and 7 ml of boron trifluoride-acetic acid complex are heated at 120° C. for 10 minutes. After cooling, the reaction mixture is treated with 50 ml of 6 N sodium hydroxide solution, the aqueous mixture is extracted with ethylacetate, dried over sodium sulfate and the solvents evaporated. The crude material is recrystallized from hot ethanol.
A solution of the hydroxy derivative (1 mmol) and corresponding isocyanate (1.1 mmol) in 10 ml abs. pyridine was refluxed and stirred for 1 hour. Then the solvent was evaporated, 10 ml 1N hydrochloric acid was added to the residue and extracted with 3×10 ml ethyl acetate. Organic phase was dried over sodium sulphate and evaporated to dryness and crystallized from isopropanol or cleaned by chromatography on silica column using ethyl acetate.
The protected derivative D198 (0.5 mmol) was dissolved in 5 ml ethyl acetate and 1 ml ethyl acetate saturated with hydrochloric acid was added with stirring. After one hour the formed crystals were filtered and washed carefully with ethyl acetate and dried in vacuum.
wherein
the substituents X1, Y1, Y2, Y3, Y4, R2, and R5 have the meaning as defined in claim 1.
10 mmol of the corresponding 2-amino derivative (ketal derivative for 6-hydroxy compound) was dissolved in 10 dry pyridine and 11 mmol chloro-oxo-acetic acid methyl ester was added dropwise with stirring at room temperature. After two hours the reaction mixture was diluted with 10 ml of water and the crystalline product was filtered. Methyl ester was removed by equivalent amount of sodium hydroxide in 50% aqueous methanol (stirring at room temperature overnight, and isolated in crystalline form when the reaction mixture was acidified with 1 N hydrochloric acid. Ketal protecting group was removed with cupric chloride as given elsewhere. 2-hydroxyethylamide was obtained from the methyl ester and aminoethanol as representative for H2N—R5 in ethanolic solution or any other suitable solvent. The product crystallizes after 2 days at room temperature.
The following compounds were synthesized according to this procedure:
To a solution of 0.427 g (1.64 mmol) 2-(Cyclopropanecarbonyl-amino)-benzo[b]thiophene-3-carboxylic acid amide in 35 ml acetic acid was added 0.195 g (2.3 mmol) NaNO3 at room temperature. It was stirring for two days. The precipitate was collected, washed with water. The yield is 65%.
0.305 g (1 mmol) 2-(Cyclopropanecarbonyl-amino)-5-nitro-benzo[b]thiophene-3-carboxylic acid amide was hydrogenated in 40 ml abs. ethanol with catalytic amount of Pd/C (atmospheric pressure). The mixture was stirred overnight at 25° C., then filtered, and the solution was evaporated to dryness to give the title compound in 90% yield.
0.195 g (0.71 mmol) 5-Amino-2-(cyclopropanecarbonyl-amino)-benzo[b]thiophene-3-carboxylic acid amide was dissolved in 5 ml abs. dichloroethane. 0.141 g (0.71 mmol) 4-bromo-phenylisocyanate was added. The mixture was stirred at 100° C. for 3 h. The reaction was followed by TLC (EtOAc:Hexan 1:1). The solution was cooled and hexane was added, the resulted precipitate was filtered off and washed with hexane. The yield is 80%
According to this procedure the following compounds were prepared:
To a solution of 1 mmol 2-[3-(substituted-phenyl)-ureido]-6-oxo-4,5,6,7-tetrahydro-benzo[b]thiophene-3-carboxylic acid amide in acetic acid (150 ml) was added 2.4 mmol bromine in 24 ml acetic acid at 50° C. The progress of the reaction was followed by TLC.(CHCl3:MeOH 10:1). When the reaction was completed, acetic acid was evaporated. The product was crystallized from isopropanol.
According to this general method the following compounds were prepared:
Biochemical Methods and Experiments
In the following documents, background information is given with regard to the methods, micoorganisms and enzymes used according to the present invention: Peirs et al., A serine/threonine protein kinase from Mycobacterium tuberculosis, Eur. J. Biochem., March 1, 244(2), 604-612 (1997); Arruda et al., Cloning of an M. tuberculosis DNA fragment associated with entry and survival inside cells, Science 261, 1454-1457 (1993); Wieles et al., Increased intracellular survival of Mycobacterium smegmatis containing the Mycobacterium leprae thioredoxin-thioredoxin reductase gene, Infect Immun. 65(7), 2537-2541 (1997); Zahrt, Mycobacterium tuberculosis signal transduction system required for persistent infections, Proc. Natl. Acad. Sci. 98 (22), 12706-12711 (2001); and Mundayoor et al., Identification of genes involved in the resistance of mycobacteria to killing by macrophages, Ann. N. Y. Acad. Sci. 730, 26-36 (1994).
Bacterial Strains and Culture Conditions
Mycobacterium smegmatis was grown in Middlebrook 7H9 medium (supplier: Difco), supplemented with 10% ADC (Difco), 0.05% Tween-80 and 0.5% glycerol. E. coli was cultivated in LB- or TB-broth without any additional ingredients. Cloning, mutagenesis and expression of PknG and other mycobacterial kinases was done as described by Koul et. al. (Serine/threonine kinases, PknG and PknF of Mycobacterium tuberculosis: characterisation and localisation. Microbiology, 147, 2001).
GST-Fusion Protein Purification
Purification of GST-fusion proteins was done as described previously by Koul et. al. (Serine/threonine kinases, PknG and PknF of Mycobacterium tuberculosis: characterisation and localisation. Microbiology, 147, 2001). E. coli BL21 cultures containing the respective plasmids were grown overnight in TB-broth. After IPTG induction, the suspensions were incubated for another 16 hours at room temperature. The bacteria were harvested by centrifugation, resuspended in 1× PBS and lysed by sonification. After addition of Triton X-100 (1% final concentration) and subsequent clarifying of the lysates the GST-fusion proteins were purified by addition of GST-sepharose following PBS washes. The proteins were eluted with a buffer containing 50 mM glutathion, 20 mM Tris (pH 8.0), 0.1 M NaCl, 0.1 M Triton X-100 and 1 mM DTT. Thereafter, the eluates were dialysed in 20 mM HEPES (pH 7.5) and 30% glycerol.
Determination of Protein Kinase Activity
The activity of all protein serine threonine kinases from Mycobacterium tuberculosis was determined by addition of myelin basic protein as a substrate in an in vitro kinase assay. The buffer conditions were as follows: 20 mM HEPES (pH 7.5), 20 mM MgCl2, and 5 mM MnCl2, for all kinases except PknG, PknI, PknJ, and PknL. These protein kinases required lower salt concentrations, namely 1 mM MgCl2, and 1 mM MnCl2. The optimum ATP concentration for each kinase was determined by titration of ATP in a range between 0.0033 μM and 100 μM. The inhibitor studies were performed with ATP concentrations similar to the Michaelis constant (Km) for ATP. We further analysed the role of PknG in pathogenesis of mycobacteria in cellular infection model.
Infection of Macrophage Cells with Recombinant Mycobacterium smegmatis
Mycobacterium smegmatis, electroporated with either vector alone or mycobacterial expression vector containing PknG (wild type) or PknG-K181M (Mutant), was cultured for 2 days in Middlebrook 7H9 medium containing 0.05% Tween-80 and 0.5% glycerol. Bacteria were pelleted at 1500×g for 3 minutes by centrifugation and resuspended by vigorous agitating (Vortex) in Dulbecco's modified Eagle's medium (DMEM, GIBCO-BRL, Gaithersburg, USA)) containing 5% fetal bovine serum (FBS) for infecting murine macrophage cell line RAW (American Type Culture Collection No. 91B-71). This yielded a bacterial supernatant consisting mostly of single mycobacterial cells as observed by acid fast staining. Under the assumption that an optical density (O.D.) of 0.1 at 650 nm equals to 108 CFU/ml (see in this respect Wei et al., “Identification of a Mycobacterium tuberculosis Gene that enhances survival of M. smegmatis in Macrophages”, J. Bacteriol. 182, 377-384 (2000)), the O.D. of Mycobacterium smegmatis cell suspension was measured and diluted to 5×106 CFU/ml in DMEM containing 5% FBS. Viable counts were performed on Middlebrook 7H10 medium.
The RAW cell line was maintained in DMEM medium supplemented with 10% FBS. The survival assay for recombinant Mycobacterium smegmatis was performed as described by Wei et al., cited above. RAW cells were plated in a 24 well tissue culture plate (4×105 cells/well) and incubated overnight in 5% CO2 at 37° C. The inoculum (1 ml) containing 5×106 recombinant Mycobacterium smegmatis was added to achieve multiplicities of infection (moi) of 10. The plate was incubated for 2 hours at 37° C. in 5% CO2. The infected monolayers were washed twice with warm DMEM and treated with 2% FCS containing 200 μg of amikacin/mI for 1 hour at 37° C. to kill extracellular M. smegmatis. The cells were again washed twice with warm DMEM and further incubated in DMEM containing 20 μg of amikacin. This time point was considered 0 hours of infection. The 24 hours infected monolayer was incubated with 20 μg of amikacin/ml to prevent extracellular growth of any bacteria released by premature lysis of infected RAW cells. Cells were washed twice with warm DMEM before lysis was effected by addition of a 0.1% SDS solution and vigorously pipeting several times to ensure lysis of cells and release of surviving bacteria. The lysates were diluted in 7H9 broth and plated onto 7H10 agar plates and CFU were counted after incubation at 37° C. for 4 to 5 days.
Validation of Mycobacterial Kinase as a Mycobacterial Virulence Gene
Mycobacterium smegmatis was electroporated either with wildtype or mutant kinase (which exerts no kinase activity) or vector control. Mouse macrophage cell line (RAW) was infected with the various recombinant M. smegmatis expressing either pknG wild type or PknG K/M mutant or vector alone. After infection, the cells were lysed at different time points and the amount of intracellular bacteria was analysed. As can be seen from
Controls:
Reaction Buffer:
20 mM Tris-HCl, pH 7.5
10 mM MgCl2
1 mM DTT
Final Assay Concentrations:
Pipetting Sequence:
Determination of RNA Polymerase II C-terminal Domain Phosphorylation:
The phosphorylation status of RNA polymerase II C-terminal domain was determined by western blot techniques. PM1 cells were seeded in 6-well plates at a density of 5×105 per well. After over night incubation cells were treated with compound as indicated in the respective experiments. Cells were pelleted and lysed with 300 μl 3× Laemmli buffer followed by 30 min denaturing at 65° C. After separation of equal lysate volumes by SDS-PAGE the proteins were transferred to nitrocellulose membranes (Schleicher&Schuell) and probed with anti-SER2 (H5), anti-SER5 (H14) or RNA Poll II-antibodies purchased from Eurogentec and Santa Cruz, respectively. The amount of reactive protein was visualized by ECL detection methods (Amersham).
Growth Assay Using Alamar Blue™:
PM1 cells were seeded in 12-well plates at a density of 1.5×105 per well with RPMI 1640 containing 10% FCS (fetal calf serum), 1% L-Glutamine and 1% Na-Pyruvate (Sigma). Cells were incubated with compound for 2-3 days (37° C., 6% CO2) followed by subsequent splitting and renewing of compound-containing medium. At each of these time points an aliquot of cells served as data point for relative growth (given in % of the DMSO control [=100%]). The cell number was determined by addition of 10 μL Alamar Blue™ (Biozol) to 100 μl cell aliquots following the manufacturer's instructions.
HIV Replication Assay in PM1 Cells:
PM1 cells were seeded in 12-well plates at a density of 1.5×105 per well with RPMI 1640 containing 10% FCS, 1% L-Glutamine and 1% Na-Pyruvate (Sigma). Cells were previously infected with HIV-1 BaL for 3 h at a concentration of about 5×108 μg p24/cell. After addition of the respective compounds cells were incubated for 6 to 10 days. During this incubation the cells were passaged and compound-containing medium was renewed. The concentration of p24 in the cellular supernatants was determined at each of this time points using a previously described ELISA assay (Bevec et al., Proceedings of the National Academy of Sciences U.S.A. 1992, 89(20), 9870-9874).
NFκB-Dependent Transcriptional Activity:
The used NIH 3T3 75E11/300D8 cell line is described elsewhere (J. Eickhoff et al., Journal of Biological Chemistry, 2004, 279(10), 9642-9652).
HBV-Replication:
To test anti-HBV-activity of compounds the HBV-producing cell line HepG2-2.2.15 (M. A. Sells, PNAS 1987, 84, 1005-1009) was used. 1.0×104 cells were seeded in 96-well microtiter plates in DMEM medium supplemented with 10% FCS. After incubation at 37° C. in 5% CO2 atmosphere for 24 hours the medium was replaced with fresh medium containing the appropriately diluted compound. 3 days later medium was replaced by freshly prepared inhibitor-containing medium and the cells were incubated for further 3 days. Subsequently 200 μl lysis buffer (50 mM Tris-Cl 7.5; 1 mM EDTA 8.0; 0.5% NP40) per well was added. To remove cell debris and nucleic acids, lysate was centrifuged (15000 rpm, 10 min, 4° C.). Cellular and viral RNA was removed by addition of 2 μl of RNase. 100 μl of the samples were spotted onto an uncharged nylon membrane pre-wetted with PBS (phosphate-buffered saline) using a 96 well-blotting chamber (MINIfold Dot-Blot, Schleicher&Schüll). After further washing with 200 μl PBS per well the membrane was treated twice with 0.5M NaOH, 1.5M NaCl (2 min) and 4 times with 0.5M Tris 7.5, 3M NaCl (1 min). The nucleic acids were fixed by UV-treatment and used for hybridisation with a radioactive HBV-fragment prepared from the overgenome-length HBV-plasmid pT-HBV1.3 (L. G. Guidotti et al., Journal of Virology 1995., 69(10), 6158-6169).
The fixed membrane was pre-hybridized in a standard hybridisation buffer (50% formamide, 5×SSPE, 10×Denhards, 1% SDS, 100 μg/ml salmon sperm DNA) for at least 3 hours at 42° C. and hybridised overnight against the labelled HBV-fragment. The preparation of the HBV-fragment with the “Random primers DNA labelling system” (Invitrogen) was done according to the manufacturer's instructions. Hybridized filter were washed at room temperature with 2×SSC, at 62° C. with 2×SSC, 0.5% SDS and at 62° C. with 0.5×SSC, 0.5% SDS. Each washing step was carried out twice. The intensity of the HBV-DNA was quantified using a phosphoimager (Fuji). To test the cell viability 0.5×104 HepG2-2.2.15-cells were seeded in 96-well-microtiter plates in DMEM medium supplemented with 10% fetal bovine serum. After incubation at 37° C. for 24 hours the medium was replaced by fresh compound-containing medium. 3 days later medium was replaced again by freshly prepared medium containing the inhibitor and the cells were incubated for further 3 days at 37° C. After the incubation period 1/10 volume of Alamar Blue (Serotec) solution containing a growth dependant indicator was added and the cells were incubated for 3 h at 37° C. Absorbance was monitored at 570 nm and 600 nm wavelength.
HCMV Replication:
Human foreskin fibroblasts (HFF) cell culture were grown in DMEM containing 10% FCS. For HCMV-replication assays, HFF cells were infected with HCMV strain AD169 producing EGFP (HCMV AD169-GFP; 27). 1 h post infection, medium was changed with medium containing the indicated compound concentration (0.3 μM, 1 μM and 3 μM, respectively) After incubation of 7 days cells were lysed (in 25 mM Tris, pH 7.5, 2 mM DTT, 1% Triton X-100 and 10% glycerol) and analysed for EGFP content in a Wallac Victor fluorescence detector.
HCV Replicon Assays:
Compounds were tested for activity in the HCV replicon system described by Bartenschlager and coworkers (Lohmann et al, Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285, 110. 1999).
{circle around (1)} Compound Number
{circle around (2)} Target AKT1/PKBa act. HIS
{circle around (3)} Target CDK1/CycB act.GST-HIS
{circle around (4)} Target CDK2/CycA
{circle around (5)} Target CK1-alpha GST
{circle around (6)} Target EGFR GST-HIS
{circle around (7)} Target GSK-3beta HIS
{circle around (8)} Target IKKbeta HIS
{circle around (9)} Target InsR GST
{circle around (10)} Target Jnk1a1 HIS
Target Kit human GST
Target PDGFRbeta GST
Target RICK-STREP 1
Target ROCK2 human HIS
Target RSK1 act. HIS
Target SRPK1 GST
Target c-Src HIS
Target cMet GST-HIS
Target p70S6K HIS
Table I shows the half-maximal inhibition concentration (IC50) values of representative compound according to general formula (I). Table II shows inhibition rates greater than 50% of various kinases. The results exhibited in both tables prove that the compounds of the present invention are potent pharmaceutically active agents against various diseases that can be treated and/or prohibited by inhibition of the targets {circle around (1)}-{circle around (10)}, -
.
| Number | Date | Country | Kind |
|---|---|---|---|
| 04012814.2 | May 2004 | EP | regional |
| 03020616.3 | Sep 2003 | EP | regional |
| 04004891.0 | Mar 2004 | EP | regional |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP04/10161 | 9/10/2004 | WO | 3/6/2007 |
| Number | Date | Country | |
|---|---|---|---|
| 60502606 | Sep 2003 | US | |
| 60551341 | Mar 2004 | US | |
| 60577043 | Jun 2004 | US |
