BENZOPYRROLIDONE DERIVATIVES POSSESSING ANTIVIRAL AND ANTICANCER PROPERTIES |
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| 申请号 | EP15155298.1 | 申请日 | 2015-02-16 | 公开(公告)号 | EP3056202A1 | 公开(公告)日 | 2016-08-17 |
| 申请人 | Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V.; Vichem Chemie Ltd.; | 发明人 | Kéri, György; Örfi, László; Horváth, Zoltán; Szokol, Bálint; Dobos, Judit; Nemes, Zoltán; Szàntay Kis, Csaba; Erös, Dánilel; Breza, Nóra; Baska, Ferenc; Karlas, Alexander; Gödert, Sigrid; Meyer, Thomas F.; | ||||
| 摘要 | The present invention relates to benzopyrrolidone derivatives and/or pharmaceutically acceptable salts thereof for the use in the treatment of infectious diseases or cancer, and pharmaceutical compositions containing at least one of said benzopyrrolidone derivatives and/or pharmaceutically acceptable salts thereof for the use in the treatment of infectious diseases or cancer. | ||||||
| 权利要求 | |||||||
| 说明书全文 | The present invention relates to benzopyrrolidone derivatives and/or pharmaceutically acceptable salts thereof for use in the treatment of infection diseases, and pharmaceutical composition containing at least one of the said benzopyrrolidone derivatives and/or pharmaceutically acceptable salt thereof. Despite vaccination and currently available antiviral drugs, influenza infections still make a huge impact on human health worldwide. In light of the risk posed by seasonal infections and also the recurring threat of influenza virus pandemics, there is an acute need to develop effective and lasting drugs. In Germany alone, the seasonal occurrence of influenza causes between 5,000 and 20,000 deaths per year (source: Robert Koch Institute). Worldwide, up to 500,000 humans die annually due to seasonal influenza or accompanying opportunistic infections (source: WHO). The recurring big influenza pandemics, however, present the greatest threat. The first great pandemic, the so-called "Spanish Flu", cost 40 million lives in the years 1918-1920, including a high percentage of healthy, middle-aged people. A similarly devastating pandemic could still be initiated by the recently arisen avian H5N1 influenza virus, if acquired mutations enable the virus to be transmitted from human-to-human. With mortality rates of approximately 60%, the H5N1 virus represents an unprecedented risk to human health. While the threat of a H5N1 pandemic still exists, the world is currently in the throes of yet another novel influenza strain, the pandemic (H1N1) 2009 virus. Although highly transmissible, the H1 N1 virus appears, currently, to be less hazardous, as mortality rates are still low (less than ∼1%) and comparable to seasonal epidemics. However, uncertainty exists about a possible acquisition of increased pathogenicity during its unavoidable adaptation process. Historical data from 1918 are giving special cause for concern: the first infections with the H1 N1 "Spanish Flu" virus in March 1918 led to relatively mild symptoms compared to the deadly second wave of infections that spread worldwide several months later. The current panel of preventive and therapeutic measures against influenza virus infections rests on (i) active vaccination and (ii) the use of conventional anti-viral drugs. Both strategies have their intrinsic limitations owing to the high variability of influenza viruses. The genome of influenza viruses consists of eight genome segments that can be quickly re-assorted upon viral co-infections, a process called antigenic shift. If such viral recombinants are derived from animal reservoirs they may exhibit surface antigens that are unknown to the human immune system. Moreover, virus replication supports the generation of point mutations, giving rise to a steady 'antigenic drift' and viral escape. These two mechanisms of influenza virus variation necessitate the generation of new anti-influenza vaccines every yearly season. However, vaccine development is inherently afflicted with uncertainty about the emerging strain because the viral antigen has to be decided upon months before the virus is established in the human population. Similarly, in the case of a pandemic threat, as the new virus has to be isolated and amplified prior to vaccine generation, a lag time of several months is unavoidable. Since pandemic strains have the potential to spread rapidly across the globe, vaccination strategies necessitate the concurrent development and availability of effective complementary options to treat already infected individuals. The fact that treatment (and immediate prophylaxis) of influenza virus infections is hampered by the occurrence of viral escape mutants is not just theoretical, but has been impressively demonstrated with the growing emergence of, e.g. Tamiflu®, resistant virus mutants of seasonal influenza strains as well as the new H5N1 and H1 N1 strains. Considering that, in the case of a severe pandemic threat, these conventional drugs would be given to millions of people; a global manifestation of drug resistance will be likely. Existing anti-viral drugs are typically directed against bona fide viral targets, so-called direct antiviral targets, bearing the usual risk of generating therapeutic resistance. However, a largely ignored fact is that viral infection and replication intimately depend on the cellular factors provided by the host. Blocking human functions to prevent infection may appear - at a first glance - counter-intuitive. However, this very approach is taken on a regular and sometimes long-lasting basis to successfully treat thousands of other human diseases. Application of this widely used approach should thus also be feasible to treat viral infection, but this approach has not been systematically developed. Recently, suitable human genes have been identified in the process of a genome-wide RNAi screen for human factors essential for influenza virus replication in vitro ( Colorectal cancer is the third most commonly diagnosed cancer and the third leading cause of cancer death in both gender. Localized tumors may be curable with surgery, but those that have spread widely are usually not curable. Therefore, there is a need for more effective treatment regimes for advanced and metastatic disease. Besides cytotoxic chemotherapy patients may take advantage of so far limited targeted therapies (e. g. anti-EGFR therapy). Many kinases have been found to be involved in the processes leading to tumor cell proliferation and survival. Benzopyrrolidone derivatives are well known kinase inhibitors (e.g. VEGFR, FGFR, PDGFR, c-KIT, FLT3, Met, Src and Aurora B). Several indolin-2-ones have been under preclinical or clinical study for cancer treatment. Benzopyrrolidone derivatives like sunitinib, nintedanib and toceranib have already been used as anticancer and anti-angiogenic agents. It is object of the present invention to provide compounds and/or pharmaceutically acceptable salts thereof, which can be used as pharmaceutically active agents, especially for treatment of infection diseases, as well as compositions comprising at least one of these compounds and/or pharmaceutically acceptable salts thereof as pharmaceutically active ingredients. This object is solved by providing compounds and/or their pharmaceutically acceptable salts according to the general formula (I) for use in the treatment of infectious diseases and pharmaceutical compositions comprising at least one of said compounds. Therefore the present invention relates to benzopyrrolidone derivatives of the general formula (I) for use in the treatment of infectious diseases, preferably in the treatment of viral infection such as influenza infection. The objective of the present invention is solved by the teaching of the independent claims. Further advantageous features, aspects and details of the invention are evident from the dependent claims, the description, the figures, and the examples of the present application. Thus, the present invention relates to compounds of general formula (I) wherein,
and stereoisomeric forms, E/Z isomers, enantiomers, mixtures of enantiomers, anomers, deoxyforms, diastereomers, mixtures of diastereomers, prodrugs, tautomers, hydrates, solvates and racemates of the above mentioned compounds and pharmaceutically acceptable salts thereof, for the use in the treatment of infectious diseases and cancer. The term 'prodrug' describes a precursor of the active ingredient containing a compound according to general formula (I) and further comprises groups which can be cleaved under physiological conditions or releases a compound according to general formula (I) under physiological conditions. The expression 'tautomer' is defined as an organic compound that is interconvertible by a chemical reaction called tautomerization. Tautomerization can be catalyzed preferably by bases or acids or other suitable compounds. Compound according to claim 1, wherein z represents 0 and Q1 is: -H. The compounds of general formula (I) are basic and form salts with various organic and inorganic acids. Examples of suitable acids for such acid addition salt formation are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, citric acid, oxalic acid, malonic acid, salicylic acid, p-aminosalicylic acid, malic acid, fumaric acid, succinic acid, ascorbic acid, maleic acid, sulfonic acid, phosphonic acid, perchloric acid, nitric acid, formic acid, propionic acid, gluconic acid, lactic acid, tartaric acid, hydroxymaleic acid, pyruvic acid, phenylacetic acid, benzoic acid, p-aminobenzoic acid, p-hydroxybenzoic acid, methanesulfonic acid, ethanesulfonic acid, nitrous acid, hydroxyethanesulfonic acid, ethylenesulfonic acid, p-toluenesulfonic acid, naphthylsulfonic acid, sulfanilic acid, camphersulfonic acid, china acid, mandelic acid, o-methylmandelic acid, hydrogen-benzenesulfonic acid, picric acid, adipic acid, D-o-tolyltartaric acid, tartronic acid, a-toluic acid, (o, m, p)-toluic acid, naphthylamine sulfonic acid, amino acids such as glycone, alanine, valine, leucine, isoleucine, serine, threonine, phenylalanine, tyrosine, tryptophane, lysine, arginine, histidine, asparaginic acid, glutamic acid, asparagines, glutamine, cysteine, methionine, proline, 4-hydroxyproline, N,N,N-trimethyllysine, 3-methylhistidine, 5-hydroxylysine, O-phosphoserine, g-carboxyglutamate, e-N-acetyllysine, w-N-methylarginine, citrulline and ornithine, and other mineral or carboxylic acids well known to those skilled in the art. The salts are prepared by contacting the free base form with a sufficient amount of the desired acid to produce a salt in the conventional manner. The free base forms may be regenerated by treating the salt with a suitable dilute aqueous base solution such as dilute aqueous sodium hydroxide, potassium carbonate, ammonia and sodium bicarbonate. The free base forms differ from their corresponding salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the salts are otherwise equivalent to their corresponding free base forms for purposes of this invention. A preferred embodiment of this invention is directed to compounds of general formula (II) wherein the residues R2, R3, Q1 - Q4, and y have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (III) wherein the residues R2, R3, Q1 - Q4, y and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (IV) wherein the residues R3, Q1 - Q4, y and z have the meanings as defined herein. The amide moiety preferably is bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (V) wherein the residues R2, R3, Q2 - Q4, y and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (VI) wherein the residues R2, R3, Q1 - Q4, and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (VII) wherein the residues R1, R3, Q1 - Q4, and y have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (VIII) wherein the residues R1, R3, Q1 - Q4, y and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (IX) wherein the residues R1, R3, Q2 - Q4, y and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (X) wherein the residues R1, R3, Q1 - Q4, and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XI) wherein the residues R1, R3, Q2 - Q4, and y have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XII) wherein the residues R1 - R3, Q2 - Q4, y and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XIII) wherein the residues R1 - R3, Q2 - Q4, and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XIV) wherein the residues R1 - R3, Q1 - Q4, and y have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XV) wherein the residues R1 - R3, Q1 - Q4, and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XVI) wherein the residues R1 - R3, and Q1 - Q4 have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XVII) wherein the residues R2, R3 and Q1 - Q4 have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XVIII) wherein the residues R3, Q1 - Q4, and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XIX) wherein the residues R1 - R3 and Q1 - Q4 have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XX) wherein the residues R3, Q1 - Q4, y and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XXI) wherein the residues R2, R3, Q1 - Q4, and z have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XXII) wherein the residues R3, Q1 - Q4, and y have the meanings as defined herein. The amide moiety is preferably preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XXIII) wherein the residues R2, R3 and Q2 - Q4 have the meanings as defined herein. The amide moiety is preferably preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XXIV) wherein the residues R3 and Q1 - Q4 have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XXV) wherein the residues R3 and Q2 - Q4 have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XXVI) wherein the residues R4, Q2 - Q4, and p have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. A preferred embodiment of this invention is directed to compounds of general formula (XXVII) wherein the residues R1, R2, and Q2 - Q4 have the meanings as defined herein. The amide moiety is preferably bound at the 5-position of the indol backbone. Another preferred embodiment of the present invention relates to compounds of the general formulas (VII)-(XVI), (XIX) and (XXVII), wherein R1 represents: -CH2CH3, -CH(CH3)2, wherein R' and R" are independently of each other selected from: -H, -F, -CH3 and -OCH3; R"' represents: -CH3; A represents: -O- or -S-. Furthermore, a preferred embodiment refers to inventive compounds of general formulas (I)-(III), (V), (VI), (IX), (XII)-(XVII), (XIX), (XXI), and (XXVII), wherein R2 represents: -H or -COCH3. Moreover, a preferred embodiment according to the invention comprises compounds of general formulas (I)-(XXV), wherein R3 represents: R4 represents: -R12, -NH2, -N(CH3)2, -N(C2H5)2, or X represents: =O, =S; R5 represents: -R6, R6 is selected form : -H, -CH3, -C2H5, -C3H7, -CH(CH3)2, -C(CH3)3-(CH2)n-CH3, -(CH2)n-C(CH3)2, -CH[(CH2)nCH3]-(CH2)m-CH3, -CH[(CH2)nCH3]-(CH2)m-C(CH3)2, -CH=CH2, -(CH2)n-CH=CH-(CH2)k-CH3, -CH≡CH, -(CH2)n-C≡C-(CH2)k-CH3, -Ph, -(CH2)n-Ph, -(CH2)n-OH, -(CH2)n-NH2, -(CH2)n-O-(CH2)k-CH3, -(CH2)n-NH(CH2)k-CH3, -(CH2)n-N(CH3)2 , -(CH2)n-N(C2H5)2 and -(CH2)n-N(CH3)(C2H5); preferably from : -H, -CH3, -C2H5, -C3H7, -CH(CH3)2, -C(CH3)3-(CH2)n-CH3, -(CH2)n-C(CH3)2, -CH=CH2, -(CH2)n-CH=CH-(CH2)k-CH3, -CH≡CH, -Ph, -(CH2)n-Ph, -(CH2)n-OH, -(CH2)n-NH2, -(CH2)n-O-(CH2)k-CH3, -(CH2)n-N(CH3)2, and -(CH2)n-N(C2H5)2; more preferably from : -H, -CH3, -C2H5, -C3H7, -CH(CH3)2, -CH=CH2,-CH≡CH, -Ph, -CH2-Ph, -CH2-OH, and -CH2-NH2; R13 represents: -OH, -Cl, R11 represents: R12 represents: R10 has the meanings as disclosed herein and preferably R10 represents hydrogen; k, m and n have the meanings as disclosed herein; and p is an integer number selected from: 0 and 1. A further preferred embodiment according to the invention refers to the compounds of the general formulas (I)-(XXVII), wherein Q3 is in para-position and Q4 in meta-position of the benzene ring. Q3 represents: -H and Q4 is selected from: -H, -F, -Cl, -CH3, or Q4 represents: -H and Q3 is selected from: -H, -F, -Cl, or -CH3. A preferred embodiment of this invention is directed to compounds of general formulas (III), (VIII), (XII), (XIV), (XV), and (XIX)-(XVII), wherein Q2 is selected from: -CH3 and -C2H5. Furthermore, a preferred embodiment refers to inventive compounds of general formulas (I), (III)-(VI), (VII)-(XIII), (XV), (XVIII), (XX), and (XXI), wherein z represents 0. A preferred embodiment of the invention comprises the compounds according to the general formula (I) as depicted in the following Table 1 and prodrugs, tautomers, hydrates, solvates of the above mentioned compounds and pharmaceutically acceptable salts thereof, for the use in the treatment of infectious diseases and cancer. The inventive compounds (I) according to the present invention can be prepared by methods known to one skilled in the art. The synthesis is preferably carried out according to the general synthetic routes, shown in scheme 1. Compound 3* can be synthesised from the commercially available carboxy-oxindoles 1* and commercially available amines 2* via amide bond formation a*. Conveniently, many amines of general formula 2* are commercially available or can be easily prepared by a person skilled in the art following known synthetic procedures that are disclosed in patent or non-patent literature. Examples of suitable amines are: (1R)-1-(4-chlorophenyl)ethanamine, (1R)-1-phenylpropan-1-amine, (1R)-1- phenylethanamine, (1S)-1-phenylethanamine, (1R)-1-(4-methoxyphenyl)-ethanamine, (1R)-1-(3-methoxyphenyl)-ethanamine, (1R)-1-cyclohexylethanamine (1R)-1-(3-chlorophenyl)-ethanamine, (1R)-1-(2-naphthyl)ethanamine, (1S)-indan-1-amine, (1R)-indan-1-amine, (1R)-N-methyl-1-phenylethanamine, (1R)-1-(4-methylphenyl)-ethanamine, (1S)-1-(4-methylphenyl)-ethanamine, (1R)-1-(4-chlorophenyl)-ethanamine, (1R)-1-(3-bromophenyl)-ethanamine, (1R)-1-(3-chlorophenyl)propan-1-amine, (1R)-1-(4-chlorophenyl)propan-1-amine, (1R)-1-(3,4-difluorophenyl)propan-1-amine, 1-(4-methoxyphenyl)propan-1-amine,, (1R)-1-(4-methoxyphenyl)propan-1-amine, (1R)-1-(4-methylphenyl)propan-1-amine, (1R)-1-(3-methylphenyl)propan-1-amine, (1R)-1-(3-methoxyphenyl)propan-1-amine, etc. Amide bond formation involves activation of the carboxylic group, followed by nucleophile attack of the amino group. Activation of the carboxylic acid includes conversion of the carboxylic acid to the corresponding active ester, carboxyl chloride or bromide, or anhydride. For facility reasons, the activation of the carboxylic acid performed in situ by treatment of the mixture of carboxylic acid and amine with an activating agent. Activating agents include carbodiimides such as DCC (N,N'-dicyclohexylcarbodiimide), DIC (N,N'-diisopropylcarbodiimide) and EDCI (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride), phosphonium salts such as BOP (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate, PyBOP (benzotriazol-1-yloxy)tripyrrolidinophosphonium 5 hexafluorophosphate, PyAOP ((7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate), BroP (bromotris(dimethylamino)phosphonium hexafluorophosphate), PyCloP (chlorotripyrrolidinophosphonium hexafluorophosphate), DEPBT (3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one), uronium and guanidinium salts such as HBTU (O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate), TBTU (O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate), HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate), HBPyU (O-(benzotriazol-1-yl)-N,N,N',N'-bis(tetramethylene)uranium hexafluorophosphate), HCTU (0-(6-chlorobenzotriazol-1-yl)-N,N,N',N'- 8 tetramethyluronium hexafluorophosphate). Amide bond formation is conducted preferably in presence of a base, such as tertiary amines including TEA (triethylamine) and (DIPEA) diisopropylethyl amine. Thus, the amide bond formation can be carried out using reagent systems such as HATU/DIPEA, HBTU/TEA or EDCI/HOBT/TEA. Compound 5* can be made from 3* with commercially available trimethoxymethyl or triethoxymethyl derivatives 4*, such as (trimethoxymethyl)benzene, (triethoxymethyl)benzene, 2-(trimethoxymethyl)pyridine, 2-(triethoxymethyl)pyridine, 3-(trimethoxymethyl)pyridine, 3-triethoxymethyl)pyridine, 4-trimethoxymethyl)pyridine, 4-triethoxymethyl)pyridine, 2-(trimethoxymethyl)thiophene, 2-(triethoxymethyl)thiophene, 3-trimethoxymethyl)thiophene, 3-(triethoxymethyl)thiophene, by reacting them via an aldol reaction b* at reflux, in acetic anhydride or in the mixture of acetic anhydride and other appropriate solvent such as THF (tetrahydrofurane), dioxane, DMF (dimethylformamide), MeOH (methanol), EtOH (ethanol), propanol, isopropanol, butanol, tert-BuOH, forming an α,β-unsaturated vinyl ether. Reacting the vinyl ether 5* with anilines and amines, in the presence of DMF, MeOH, EtOH, DMSO (dimethyl sulfoxid), or in their appropriate mixtures, via reaction c* gives the inventive compounds (I). Anilines and amines are commercially available or can be easily prepared by a person skilled in the art following known synthetic procedures that are disclosed in patent or non-patent literature. Some examples for the anilines and amines are: 4-[(dimethylamino)methyl]aniline, 3-[(dimethylamino)methyl]aniline, 4-[(diethylamino)methyl]aniline, 3-[(diethylamino)methyl]aniline, 4-(piperidin-1-ylmethyl)aniline, 3-(piperidin-1-ylmethyl)aniline, 2-(piperidin-1-ylmethyl)aniline, 4-(pyrrolidin-1-ylmethyl)aniline, 3-(pyrrolidin-1-ylmethyl)aniline, 2-(pyrrolidin-1-ylmethyl)aniline, 4-[(4-methylpiperazin-1-yl)methyl]aniline, 3-[(4-methylpiperazin-1-yl)methyl]aniline, 2-[(4-methylpiperazin-1-yl)methyl]aniline, 2-[4-(3-aminobenzyl)piperazin-1-yl]ethanol, 2-[4-(3-aminobenzyl)piperazin-1-yl]ethanol, 2-[4-(4-aminobenzyl)piperazin-1-yl]ethanol, 2-[4-(2-aminobenzyl)piperazin-1-yl]ethanol, 4-(morpholin-4-ylmethyl)aniline, 3-(morpholin-4-ylmethyl)aniline, 2-(morpholin-4-ylmethyl)aniline, [1-(4-aminobenzyl)piperidin-2-yl]methanol, [1-(3-aminobenzyl)piperidin-2-yl]methanol, [1-(2-aminobenzyl)piperidin-2-yl]methanol, 3-(thiomorpholin-4-ylmethyl)aniline, 2-(thiomorpholin-4-ylmethyl)aniline, 4-(thiomorpholin-4-ylmethyl)aniline, N-(4-aminophenyl)-2-piperidin-1-ylacetamide, N-(3-aminophenyl)-2-piperidin-1-ylacetamide, N-(2-aminophenyl)-2-piperidin-1-ylacetamide, N-(3-aminophenyl)-N-methyl-2-piperidin-1-ylacetamide, N-(4-aminophenyl)-N-methyl-2-piperidin-1-ylacetamide, N-(2-aminophenyl)-N-methyl-2-piperidin-1-ylacetamide, N-(4-aminophenyl)-N-methyl-2-(4-methylpiperazin-1-yl)acetamide, N-(3-aminophenyl)-N-methyl-2-(4-methylpiperazin-1-yl)acetamide, N-(2-aminophenyl)-N-methyl-2-(4-methylpiperazin-1-yl)acetamide, N-(3-aminophenyl)-2-[4-(2-hydroxyethyl)piperazin-1-yl]-N-methylacetamide, N-(4-aminophenyl)-2-[4-(2-hydroxyethyl)piperazin-1-yl]-N-methylacetamide, N-(3-aminophenyl)-N-methyl-2-pyrrolidin-1-ylacetamide, N-(4-aminophenyl)-N-methyl-2-pyrrolidin-1-ylacetamide, N-(4-aminophenyl)-2-pyrrolidin-1-ylacetamide, etc. 1-[2-(aminomethyl)phenyl]-3-ethylurea, 1-[2-(aminomethyl)phenyl]-3-phenylurea, 1-[2-(aminomethyl)phenyl]-3-(3-chlorophenyl)urea, 1-[2-(aminomethyl)phenyl]-3-(4-hydroxyphenyl)urea, 1-[3-(aminomethyl)phenyl]-3-ethylurea, 1-[3-(aminomethyl)phenyl]-3-phenylurea, 1-[3-(aminomethyl)phenyl]-3-(3-chlorophenyl)urea, 1-[3-(aminomethyl)phenyl]-3-(4-hydroxyphenyl)urea, 1-[3-(aminomethyl)phenyl]-3-(1,3-benzodioxol-5-yl)urea, 1-[4-(aminomethyl)phenyl]-3-ethylurea, 1-[4-(aminomethyl)phenyl]-3-phenylurea, 1-[4-(aminomethyl)phenyl]-3-(3-chlorophenyl)urea, 1-[4-(aminomethyl)phenyl]-3-(4-hydroxyphenyl)urea, 1-[2-(2-aminoethyl)phenyl]-3-ethylurea, 1-[2-(2-aminoethyl)phenyl]-3-phenylurea, 1-[2-(2-aminoethyl)phenyl]-3-(3-chlorophenyl)urea, 1-[2-(2-aminoethyl)phenyl]-3-(4-hydroxyphenyl)urea, 1-[3-(2-aminoethyl)phenyl]-3-ethylurea, 1-[3-(2-aminoethyl)phenyl]-3-phenylurea, 1-[3-(2-aminoethyl)phenyl]-3-(3-chlorophenyl)urea, 1-[3-(2-aminoethyl)phenyl]-3-(4-hydroxyphenyl)urea, 1-[3-(2-aminoethyl)phenyl]-3-(1,3-benzodioxol-5-yl)urea, 1-[4-(2-aminoethyl)phenyl]-3-ethylurea, 1-[4-(2-aminoethyl)phenyl]-3-phenylurea, 1-[4-(2-aminoethyl)phenyl]-3-(3-chlorophenyl)urea, 1-[4-(2-aminoethyl)phenyl]-3-(4-hydroxyphenyl)urea, (3S)-quinuclidin-3-amine, (3R)-quinuclidin-3-amine. The order of reaction a* and b* during the synthesis of 5* is reversible, as shown in scheme 1. If one would chose the other synthetic route, than one should react 1*(oxindole carboxylic acid or oxindole carboxylic acid ester) with 4* in reaction b* first, gaining compound 6*. Reacting the vinyl ether 6* with anilines and amines in the presence of DMF, MeOH, EtOH, DMSO (dimethyl sulfoxid), or in their appropriate mixtures, via reaction c* gives 5B. In addition, the inventive compounds (I) according to the present invention can also (but not limited to) be prepared according to the general synthetic routes, shown in Scheme 2. Compound 11* can be synthesised from the commercially available carboxy-oxindoles (acid or ester) 10* according to procedure A. In procedure B or B2 (aldol-reaction) compound 12a,b* can be prepared from 10* and 11* reacted with commercially avaible acyl-derivatives, especially acyl-chlorides: isonicotinoyl chloride, nicotinoyl chloride, thiophene-3-carbonyl chloride, thiophene-2-carbonyl chloride, 1-methyl-1H-pyrazole-4-carbonyl chloride, 1-methyl-1H-indole-5-carbonyl chloride, 3-furoyl chloride, 2-furoyl chloride, cyclohexanecarbonyl chloride, cyclopropanecarbonyl chloride, propanoyl chloride, 2-methylpropanoyl chloride, etc. in the presence of N,N-dimethylpyridin-4-amine and TEA or DIEA in any aprotic solvents, especially dichloromethane, chloroform or dichororoethane. Compounds 12b* were obtained via procedure B2, where 2 molequivalent activated acyl-derivatives are needed for simultenously protection of acyl NH. Compound 13A* and 13B* can be obtained by activation of vinyl-OH and reacted by amines (N-C bond formation) in procedure c*. Amide bond formation involves activation of the hydroxy group by known methods via silylation, especially chlorotrimethylsilane, bromotrimethylsilane, iodotrimethylsilane, via mesylation by methanesulfonyl chloride or activation by trifluoromethanesulfonic anhydride in any aprotic solvent like dioxane, tetrahydrofurane. In some cases under condition of this reaction, N-acyl groups were cleaved from the oxindole NH, giving the desired 14* compounds. In other cases, N-acyl type protecting groups must be cleaved under acidic (especially aqueous HCl) or basic (especially aqueous LiOH, NaOH, or KOH) conditions (procedure D) In procedure E the activation of the carboxylic acid performed in situ by treatment of the mixture of the amine and compound 14* with an activating agents, such as DCC (N,N'-dicyclohexylcarbodiimide), DIC (N,N'-diisopropylcarbodiimide) and EDCI (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride), phosphonium salts such as BOP (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate, PyBOP (benzotriazol-1-yloxy)tripyrrolidinophosphonium 5 hexafluorophosphate, PyAOP ((7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate), BroP (bromotris(dimethylamino)phosphonium hexafluorophosphate), PyCloP (chlorotripyrrolidinophosphonium hexafluorophosphate), DEPBT (3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one), uronium and guanidinium salts such as HBTU (O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate), TBTU (O-(Benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate), HATU (O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate), HBPyU (O-(benzotriazol-1-yl)-N,N,N',N'-bis(tetramethylene)uranium hexafluorophosphate), HCTU (O-(6-chlorobenzotriazol-1-yl)-N,N,N',N'- 8 tetramethyluronium hexafluorophosphate). Amide bond formation is conducted preferably in presence of a base, such as tertiary amines including TEA (triethylamine) and (DIPEA) diisopropylethyl amine. Thus, the amide bond formation can be carried out using reagent systems such as HATU/DIPEA, HBTU/TEA or EDCI/HOBT/TEA. Acylation of NH of the benzopyrrolidones was carried out by acylation agents (especially acetic-anhydride) at 80 °C - 140 °C obtained the compounds covered by general formula (I). The present invention refers further to use of the compounds according to the general formula (I) are in the manufacture of a medicament for the treatment of infectios diseases. Another aspect of the present invention relates to a method comprising administering to a patient suffering from an infectious disease a therapeutically effective amount of at least one compound according to general formula (I) as defined herein in order to treat that infectious disease. As used herein the term infectious diseases comprises selected from the group comprising or consisting of virally induced infectious diseases, bacterially induced infectious diseases, and parasitic induced infectious diseases. Within the present invention it is preferred that the infectious diseases are virally induced infectious diseases, including opportunistic diseases. A preferred embodiment of the present invention refers to the compounds of general formula I for use in the treatment of virally induced infectious diseases, including opportunistic diseases, being infections with dsDNA viruses, ssDNA viruses, (+)ssRNA viruses, (-)ssRNA viruses, ssRNA-RT viruses, or dsDNA-RT viruses. Within the present invention it is preferred that the infection with (-)ssRNA viruses are an infection with Orthomyxoviridae, Bornaviridae, Filoviridae, Paramyxoviridae, or Rhabdoviridae. Another aspect of the present invention refers to the compounds of general formula I for use in the treatment of an infection with Orthomyxoviridae selected from the group comprising or consisting of an infection with Influenza virus A, Influenza virus B, or Influenza virus C. Especially preferred are compounds of general formula I for use in the treatment of an infection with Influenza virus A. The term "opportunistic diseases" as used herein refers to infections caused by pathogens, particularly opportunistic pathogens that usually do not cause disease or symptoms in a healthy host, one with a healthy immune system but in a host with a damaged or weakened immune system. This is because a healthy immune system is able to successfully fight off the disease, or keep it under control. Examples for opportunistic diseases are caused by an infection with aspergillus sp., candida albicans, cytomegalovirus, polyomavirus, kaposi's sarcoma caused by human herpesvirus 8 (HHV8), pseudomonas aeruginosa, staphylococcus aureus, streptococcus pneumonia and toxoplasma gondii. Another preferred embodiment of the present invention refers to compounds of the general formula (I) for the use in treatment of cancer, wherein cancer is selected from the group consisting of adenocarcinoma, choroidal melanoma, acute leukemia, acoustic neurinoma, ampullary carcinoma, anal carcinoma, astrocytoma, basal cell carcinoma, pancreatic cancer, desmoid tumour, bladder cancer, bronchial carcinoma, non-small cell lung cancer, breast cancer, Burkitt's lymphoma, corpus cancer, 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, brain metastases, testicle cancer, hypophysis tumor, carcinoids, Kaposi's sarcoma, laryngeal cancer, germ cell tumor, bone cancer, colorectal carcinoma, head and neck tumors, colon carcinoma, craniopharyngiomas, oral cancer, cancer of the central nervous system, liver cancer, liver metastases, leukemia, eyelid tumor, lung cancer, lymph node cancer, lymphomas, stomach cancer, malignant melanoma, malignant neoplasia, malignant tumors gastrointestinal tract, breast carcinoma, rectal cancer, medulloblastomas, melanoma, meningiomas, Hodgkin's disease, nasal cancer, neurinoma, neuroblastoma, kidney cancer, renal cell carcinomas, non-Hodgkin's lymphomas, oligodendroglioma, esophageal carcinoma, osteosarcomas, ovarial carcinoma, pancreatic carcinoma, penile cancer, plasmocytoma, prostate cancer, pharyngeal cancer, rectal carcinoma, retinoblastoma, vaginal cancer, thyroid carcinoma, esophageal cancer, spinalioms, T-cell lymphoma, thymoma, tube carcinoma, eye tumours, urethral cancer, urologic tumours, urothelial carcinoma, vulva cancer, wart appearance, soft tissue tumours, soft tissue sarcoma, Wilm's tumour, cervical carcinoma and tongue cancer. A more preferred embodiment of the present invention refers to compounds of the general formula (I) for the use in treatment of cancer, wherein the cancer is a colorectal cancer. Still another aspect of the present invention deals with pharmaceutical compositions comprising at least one compound according to general formula (I) as an active ingredient, together with at least one pharmaceutically acceptable carrier, excipient and/or diluents. The pharmaceutical compositions of the present invention can be prepared in a conventional solid or liquid carrier or diluents and a conventional pharmaceutically-made adjuvant at suitable dosage level in a known way. The preferred preparations are adapted for oral application or for injection. These administration forms include, for example, liquids, pills, tablets, film tablets, coated tablets, capsules, liposomal formulations, micro- and nano-formulations, 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 as described herein and/or a pharmaceutical acceptable salt thereof as active ingredient. The pharmaceutical compositions according to the present invention containing at least one compound as described in the present invention, especially one pure optical isomer, and/or a pharmaceutically 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, aerosol preparations consistent with conventional pharmaceutical practices. Other suitable formulations are gels, elixirs, dispersable granules, syrups, suspensions, creams, lotions, solutions, emulsions, suspensions, dispersions, 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. As pharmaceutically acceptable carrier, excipient and/or diluents can be used carriers such as preferably with an inert carrier like lactose, starch, sucrose, cellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, talc, mannitol, ethyl alcohol (liquid filled capsules). Suitable binders include starch, gelatin, natural sugars, corn sweeteners, natural and synthetic gums such as acacia, sodium alginate, carboxymethylcellulose, polyethylene glycol and waxes, 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. Suitable lubricants are selected from the group comprising boric acid, sodium benzoate, sodium acetate, sodium chloride, 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. Disintegrating agents (disintegrates) such as starch, methylcellulose, guar gum, 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 may be added. Further suitable additives are coloring agents, sweetening agents, flavoring agents, preservatives, glidents such as silicon dioxide and talc, and adsorbent such as clay, aluminum oxide. Suitable diluents are water or water/propylene glycol solutions for parenteral injections, juice, sugars such as lactose, sucrose, mannitol, and sorbitol, starches derived from wheat, corn rice, and potato, and celluloses such as microcrystalline cellulose. Compounds of general formula (I) may be administered as the sole active agent or in combination with one or more additional therapeutic agents, wherein the combination causes no unacceptable adverse effects. For example, a compound of general formula (I) and an active agent may be administered to the patient together in a single oral dosage composition such as a tablet or capsule, or each agent may be administered in separate dosage formulations. Where separate dosage formulations are used, the compound of general formula (I) and one or more additional therapeutic agents may be administered at essentially the same time (e.g., concurrently) or at separately staggered times (e.g., sequentially). In particular, the compounds of the present invention may be used in fixed or separate combination with other anti-viral agents such as interferons, acyclovir, neuraminidase inhibitors (oseltamivir and zanamivir) and M2 protein inhibitors (rimantadine). Thus, another aspect of the present invention relates to drug combinations comprising at least one compound according to general formula (I) and/or pharmaceutically acceptable salts thereof together with at least one additional anti-viral agent, especially at least one of the active agents mentioned above.
The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. Further modifications and alternative embodiments of various aspects of the invention will be apparent to those skilled in the art in view of this description. Accordingly, this description is to be construed as illustrative only and is for the purpose of teaching those skilled in the art the general manner of carrying out the invention. It is to be understood that the forms of the invention shown and described herein are to be taken as examples of embodiments. Elements and materials may be substituted for those illustrated and described herein, parts and processes may be reversed, and certain features of the invention may be utilized independently, all as would be apparent to one skilled in the art after having the benefit of this description of the invention. Changes may be made in the elements described herein without departing from the spirit and scope of the invention as described in the following claims. All reagents were commercial grade and were used as received without further purification, unless otherwise specified. Reagent grade solvents were used in all other cases, unless otherwise specified, respectively. Silica gel generally used for column chromatography was Fisher silica gel (60A, 35-70 micron). Thin layer chromatography was carried out using pre-coated silica gel F-254 plates (thickness 0.25 mm). The 1H-NMR spectra were recorded on a Bruker Avance 300 NMR spectometer controlled by TopSpin software package in deuterodimethylsulfoxide (DMSO-d6) solution at 30 °C. The data were processed by TopSpin software. The chemical shifts are referred to tetramethylsilane (δTMS = 0 ppm).The chemical shifts are expressed in ppm using the residual solvent as internal standard. Splitting patterns are designated as s (singlet), d (doublet), dd (double-doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), and bs (broad singlet). Electrospray MS spectra were obtained on a Waters LC-MS system Acquity SQD detector, Waters 2795 Alliance HPLC equipped with Waters 996 PDA Detector (HPLC column: Waters XBridge, RP C18, 3.5 µm, 4.6 mm x 50 mm; gradient MeCN/H2O, containing 0.1% HCOOH: 5% MeCN (0.5 min), 5% to 95% MeCN (5 min), 95% MeCN (0.5 min); flow rate: 2.0 ml/min. A mixture of 2-oxoindoline-5-carboxilic acid (5.221 g, 29.50 mmol) and triethylorthobenzoate (20.16 g, 90 mmol) in acetic anhydride (75 ml) were stirred at 100 °C for 8 h.The solvent was removed under reduced pressure. The residue was stirred with i-Pr2O (250 ml) for 2 h to give a solid which was filtered and washed with i-Pr2O to provide (3Z)-1-acetyl-3-[ethoxy(phenyl)methylene]-2-oxoindoline-5-carboxylic acid (3.5 g, 33 %) 1H-NMR in DMSO-d6 12.3(bs,1H); 8.61(d,1H); 8.22(d,1H); 7.90(dd,1H); 7.54(m,5H); 3.99(q,2H); 2.45(s,3H); 1.35(t,3H). tR: 3.82 min, MS(ESI):m/z (M+H)+352, (M+H)- 350. The following compound was obtained according to the procedure described by example1, step A: Starting from 2-oxoindoline-6-carboxilic acid and triethylorthobenzoate compound 2 was obtained (3.8 g, 36 %). C28H27N3O3, Mw calc: 453.2 A mixture of compound (3Z)-1-acetyl-3-[ethoxy(phenyl)methylene]-2-oxoindoline-5-carboxylic acid (4.35g ,12.39 mmol) and 3-(piperidine-1-ylmethyl)aniline (2.47 g, 13 mmol) in dry DMF were stirred at 100 C for 6 h. After cooling down the reaction mixture, MeOH (70 ml) and NaOMe (30 % wt solution in MeOH, 5 ml) were added and the reaction mixture was further stirred overnight. The resulting solid was filtered and washed with cold MeOH to provide (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene) indoline-5-carboxylic acid (3.27 g, 58 %) 1H-NMR in DMSO-d6 δ: 12.01 (s, 1 H), 10.71 (s, 1H), 7.55 (dd, J = 7.9 and 1.5 Hz, 1 H), 7.47-7.54 (ovl. m, 3H), 7.44 (m, 2H), 7.08 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.89 (dm, J = 7.6 Hz, 1 H), 6.78 (d, J = 1.5 Hz, 1 H), 6.85 (d, J = 8.1 Hz, 1 H), 6.68 (dm, J = 7.8 Hz, 1 H), 6.61 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 3.18 (s, 2H), 2.10 (m, 4H), 1.42 (m, 4H), 1.35 (m, 2H) tR: 2.51 min; MS(ESI): m/z (M+H)+454; (M+H)- 452. The following compounds were obtained according to the procedure described by example 1, step B: Compound 4 (3.17 g, 56%) was obtained satrting from (3Z)-1-acetyl-3-[ethoxy(phenyl)methylene]-2-oxoindoline-5-carboxylic acid and 4-(piperidine-1-ylmethyl)aniline:
Compound 5 (3.24 g, 55 %) was obtained from (3Z)-1-acetyl-3-[ethoxy(phenyl) methylene]-2-oxoindoline-5-carboxylic acid and 4-[(4-methylpiperazin-1-yl)methyl]aniline:
Compound 6: (3Z)-3-[({3-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(phenyl) methylene]-2-oxoindoline-5-carboxylic acid C28H28N4O3, Mw calc: 468.2 MS(ESI): m/z (M+H)+469 Compound 7: (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino} methylene)indoline-6-carboxylic acid C28H27N3O3, Mw calc: 453.2 MS(ESI): m/z (M+H)+454 Compound 8: (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino} methylene)indoline-6-carboxylic acid C28H27N3O3, Mw calc: 453.2 MS(ESI): m/z (M+H)+454 Compound 9: (3Z)-3-[({3-[(4-methylpiperazin-1-yl)methyl]phenyl}amino) (phenyl)methylene]-2-oxoindoline-6-carboxylic acid C28H28N4O3, Mw calc: 468.2 MS(ESI): m/z (M+H)+469; Compound 10: (3Z)-3-{[(4-{[4-(2-hydroxyethyl)piperazin-1-yl]methyl}phenyl)amino](phenyl)methylene}-2-oxoindoline-5-carboxylic acid C29H30N4O4 MW cal.:498.2 MS(ESI): m/z (M+H)+499 Compound 11: (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino) (phenyl)methylene]-2-oxoindoline-6-carboxylic acid C28H28N4O3, Mw calc: 468.2 MS(ESI): m/z (M+H)+469; C37H38N4O2, Mw calc.: 570.3 A solution of (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino} methylene) indoline-5-carboxylic acid (0.113 g, 0.25 mmol), HBTU (0.113 g, 0.30mmol), DIEA (0.116g, 0.9 mmol) and (1 R)-1-phenylpropane-1-amine (0.040g, 0.30 mmol) in dry DMF (3 ml) were stirred overnight. The reaction mixture was poured into saturated sodium carbonate solution((100 ml),) and extracted with EtOAc (3X30 ml).Washed with brine,dried(MgSO4), filtered and concentrated in vacuo. Residue was chromatographed on silica gel eluted with CHCl3-MeOH-TEA(15:1:0.1) provide the . (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl] amino}methylene)-N-[(1R)-1-phenylpropyl]indoline-5-carboxamide (0.62 mg, 44 %) 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.94 (s, 1 H), 8.05 (d, J = 8.5 Hz, 1 H), 7.50-7.57 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.6 Hz, 1 H), 7.46 (m, 2H), 7.23-7.35 (ovl. m, 4H), 7.20 (m, 1 H), 7.13 (dd, J = 7.8 and 7.5 Hz, 1 H), 6.95 (dm, J = 7.5 Hz, 1 H), 6.89 (d, J = 8.1 Hz, 1 H), 6.77 (dm, J = 7.8 Hz, 1 H), 6.71 (m, 1 H), 6.47 (d, J = 1.6 Hz, 1 H), 4.73 (ddd, J = 8.5, 7.3 and 7.3 Hz, 1 H), 3.30 (s, 2H), 2.14 (m, 4H), 1.61-1.81 (m, 2H), 1.45 (m, 4H), 1.36 (m, 2H), 0.81 (t, J = 7.2 Hz, 3H) tR: 3.3 min; MS(ESI): m/z (M+H)+571; (M+H)-569. The following compounds were obtained according to the procedure described by example 1, step C: Compound 13 (64 mg, 46%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene) indoline-5-carboxylic acid and (1R)-1-phenylethanamine C36H36N4O2, Mw calc, : 556.2 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.12 (d, J = 7.8 Hz, 1 H), 7.50-7.55 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.20 (m, 1 H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.93 (dm, J = 7.6 Hz, 1 H), 6.88 (d, J = 8.1 Hz, 1 H), 6.76 (dm, J = 7.8 Hz, 1 H), 6.69 (m, 1 H), 6.46 (d, J = 1.5 Hz, 1 H), 4.98 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.22 (s, 2H), 2.11 (m, 4H), 1.43 (m, 4H), 1.36 (d, J = 7.0 Hz, 3H), 1.35 (m, 2H), tR: 3.17 min; MS(ESI): m/z (M+H)+557; (M+H)-555. Compound 14 (64 mg, 45%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene) indoline-5-carboxylic acid and (1S)-1-phenylpropan-1-amine C37H38N4O2, Mw calc. : 570.3 1H-NMR in DMSO-d6 δ: 12.04 (s, 1 H), 10.94 (s, 1 H), 8.06 (d, J = 8.5 Hz, 1 H), 7.50-7.55 (ovl. m, 3H), 7.49 (dd, J = 8.2 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.20 (m, 1 H), 7.12 (dd, J = 7.8 and 7.7 Hz, 1 H), 6.93 (dm, J = 7.7 Hz, 1 H), 6.89 (d, J = 8.2 Hz, 1 H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.68 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 6.47 (d, J = 1.5 Hz, 1 H), 4.73 (ddd, J = 8.5, 7.2 and 7.2 Hz, 1 H), 3.21 (s, 2H), 2.10 (m, 4H), 1.60-1.80 (m, 2H), 1.43 (m, 4H), 1.34 (m, 2H), 0.81 (t, J = 7.2 Hz, 3H) tR: 3.29 min; MS(ESI): m/z (M+H)+571; (M+H)-569. Compound 15 (57 mg, 38%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene) indoline-5-carboxylic acid and (1S)-1-(4-chlorophenyl)ethanamine C36H35ClN4O2 Mw calc.: 590.2 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.94 (s, 1 H), 8.17 (d, J = 7.8 Hz, 1 H), 7.49-7.56 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.35 (dm, J = 8.5 Hz, 2H), 7.30 (dm, J = 8.5 Hz, 2H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.92 (dm, J = 7.6 Hz, 1 H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.68 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 6.45 (d, J = 1.5 Hz, 1 H), 4.96 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.20 (s, 2H), 2.10 (m, 4H), 1.40 (m, 4H), 1.35 (d, J = 7.0 Hz, 3H), 1.34 (m, 2H) tR: 3.37 min; MS(ESI): m/z (M+H)+591; (M+H)-589. Compound 16 (69 mg, 46%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene) indoline-5-carboxylic acid and (1R)-1-(4-chlorophenyl)ethanamine C36H35ClN4O2, Mw calc. : 590.2 1H-NMR in DMSO-d6 δ: 12.02 (s, 1 H), 10.94 (s, 1 H), 8.16 (d, J = 7.8 Hz, 1 H), 7.49-7.56 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.35 (dm, J = 8.5 Hz, 2H), 7.30 (dm, J = 8.5 Hz, 2H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.92 (dm, J = 7.6 Hz, 1 H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.68 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 6.45 (d, J = 1.5 Hz, 1 H), 4.96 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.20 (s, 2H), 2.10 (m, 4H), 1.40 (m, 4H), 1.35 (d, J = 7.0 Hz, 3H), 1.34 (m, 2H) tR: 3.35 min; MS(ESI): m/z (M+H)+591; (M+H)-589. Compound 17 (65 mg, 45%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene) indoline-5-carboxylic acid and (1R)-1-(4-fluorophenyl)ethanamine C36H35FN4O2, Mw calc. : 574.2 1H-NMR in DMSO-d6 δ: 12.02 (s, 1 H), 10.94 (s, 1 H), 8.14 (d, J = 7.8 Hz, 1 H), 7.49-7.57 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.31 (ddm, J = 8.6 and 5.6 Hz, 2H), 7.12 (ddm, J = 9.0 and 8.6 Hz, 2H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.93 (dm, J = 7.6 Hz, 1 H), 6.88 (d, J = 8.1 Hz, 1 H), 6.76 (dm, J = 7.8 Hz, 1 H), 6.69 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 6.46 (d, J = 1.5 Hz, 1 H), 4.98 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.21 (s, 2H), 2.10 (m, 4H), 1.42 (m, 4H), 1.35 (d, J = 7.0 Hz, 3H), 1.34 (m, 2H) tR: 3.16 min; MS(ESI): m/z (M+H)+575; (M+H)-573. Compound 18 (65 mg, 45%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine C37H38N4O2, Mw calc.: 570.3 1H-NMR in DMSO-d6 δ: 12.04 (s, 1 H), 10.92 (s, 1 H), 8.04 (d, J = 8.3 Hz, 1 H), 7.49-7.57 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.21 (m, 1 H), 7.04 (dm, J = 8.2 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 8.2 Hz, 2H), 6.46 (d, J = 1.5 Hz, 1 H), 4.73 (ddd, J = 8.3, 7.3 and 7.3 Hz, 1 H), 3.30 (s, 2H), 2.23 (m, 4H), 1.60-1.80 (m, 2H), 1.44 (m, 4H), 1.36 (m, 2H), 0.81 (t, J = 7.2 Hz, 3H) tR: 3.26 min; MS(ESI): m/z (M+H)+571; (M+H)-569 Compound 19 (75 mg, 52%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-N-methyl-1-phenylethanamine C37H38N4O2, Mw calc.: 570.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.88 (s, 1 H), 7.42-7.58 (ovl. m, 5H), 7.37 (ddm, J = 7.5 and 7.0 Hz, 2H), 7.38 (tm, J ∼ 7.0 Hz, 1H), 7.13 (dm, J ∼ 7.5 Hz, 2H), 7.11 (dd, J = 7.8 and 7.6 Hz, 1 H); 7.03 (dd, J = 8.00 and 1.3 Hz, 1 H), 6.92 (dm, J = 7.6 Hz, 1 H), 6.91 (d, J = 8.0 Hz, 1 H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.70 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 5.81 (d, J ∼ 1.5 Hz, 1 H), 5.40 (br, 1 H), 3.20 (br. s, 2H), 2.36 (s, 3H), 2.10 (m, 4H), 1.43 (m, 4H), 1.39 (d, J ∼ 7.0 Hz, 3H), 1.35 (m, 2H) tR: 3.21 min ; MS(ESI): m/z (M+H)+ 571; (M+H)-569 Compound 20 (70 mg, 51%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and 1-phenylmethanamine C35H34N4O2, Mw calc.: 542.3 1H-NMR in DMSO-d6 δ: 12.04 (s, 1 H), 10.93 (s, 1 H), 8.42 (t, J = 5.9 Hz, 1 H), 7.50-7.60 (ovl. m, 3H), 7.47 (m, 2H), 7.46 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.31 (ddm, J ∼ 7.5 and 7.0 Hz, 2H), 7.16-7.26 (ovl. m, 3H), 7.05 (dm, J = 8.2 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 8.2 Hz, 2H), 6.53 (d, J ∼ 1.0 Hz, 1 H), 4.32 (d, J = 5.9 Hz, 2H), 3.28 8s, 2H), 2.22 (m, 4H), 1.44 (m, 4H), 1.36 (m, 2H) tR: 3.03 min ; MS(ESI): m/z (M+H)+543; (M+H)-541 Compound 21 (60 mg, 41%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-(4-methoxyphenyl)ethanamine C37H38N4O3, Mw calc. : 586.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.92 (s, 1 H), 8.02 (d, J = 8.0 Hz, 1 H), 7.50-7.57 (ovl. m, 3H), 7.48 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.45 (m, 2H), 7.19 (dm, J = 8.6 Hz, 2H), 7.04 (dm, J = 8.3 Hz, 2H), 6.87 (d, J = 8.1 Hz, 1 H), 6.86 (dm, J = 8.6 Hz, 2H), 6.75 (dm, J = 8.3 Hz, 2H), 6.44 (d, J = 1.5 Hz, 1 H), 4.94 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.72 (s, 3H), 3.29 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.36 (m, 2H), 1.34 d, J = 7.0 Hz, 3H) tR: 3.08 min; MS(ESI): m/z (M+H)+587; (M+H)-585 Compound 22 (67 mg, 46%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-(3-methoxyphenyl)ethanamine C37H38N4O3, Mw calc.: 586.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.92 (s, 1 H), 8.09 (d, J = 8.0 Hz, 1 H), 7.50-7.55 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.21 (dd, J = 8.0 and 7.8 Hz, 1 H), 7.05 (dm, J = 8.3 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.87 (dd, J ∼ 1.5 and 1.5 Hz, 1 H), 6.85 (dm, J = 7.8 Hz, 1 H), 6.78 (dm, J = 8.0 Hz, 1 H); 6.75 (dm, J = 8.3 Hz, 2H), 6.44 (d, J = 1.0 Hz, 1 H), 4.95 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.73 (s, 3H), 3.28 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.36 (m, 2H), 1.35 d, J = 7.0 Hz, 3H) tR: 3.11 min; MS(ESI): m/z (M+H)+587; (M+H)-585 Compound 23 (71 mg, 49%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-(4-fluorophenyl)ethanamine C36H35FN4O2, Mw calc.: 574.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.92 (s, 1 H), 8.18 (d, J = 8.0 Hz, 1 H), 7.50-7.55 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.31 (ddm, J = 8.6 and 5.5 Hz, 2H), 7.12 (ddm, J = 9.1 and 8.6 Hz, 2H), 7.05 (dm, J = 8.3 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 8.3 Hz, 2H), 6.45 (d, J = 1.0 Hz, 1 H), 4.98 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.28 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.36 (m, 2H), 1.35 d, J = 7.0 Hz, 3H) tR: 3.16 min ; MS(ESI): m/z (M+H)+575; (M+H)-573 Compound 24 (57 mg, 39%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-(3-methoxyhenyl)ethanamine C37H38N4O3, Mw calc.: 586.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.09 (d, J = 7.8 Hz, 1 H), 7.50-7.55 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.21 (dd, J = 8.0 and 7.5 Hz, 1 H), 7.11 (dd, J = 7.8 and 7.8 Hz, 1 H), 6.92 (dm, J = 7.8 Hz, 1 H), 6.88 (d, J = 8.1 Hz, 1 H), 6.86 (dd, J = 1.5 and 1.5 Hz, 1 H), 6.85 (dm, J = 8.0 Hz, 1 H), 6.78 (dm, J = 7.5 Hz, 1 H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.69 (dd, J ∼ 1.5 and 1.5 Hz, 1 H), 6.45 (d, J = 1.0 Hz, 1 H), 4.95 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.73 (s, 3H), 3.20 (s, 2H), 2.10 (m, 4H), 1.42 (m, 4H), 1.35 d, J = 7.0 Hz, 3H)1.34 (m, 2H), tR: 3.13 min ; MS(ESI): m/z (M+H)+587; (M+H)-585 Compound 25 (51 mg, 37%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1S)-1-phenylethanamine C36H36N4O2, Mw calc. : 556.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.12 (d, J = 7.8 Hz, 1 H), 7.50-7.55 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.20 (m, 1 H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.93 (dm, J = 7.6 Hz, 1 H), 6.88 (d, J = 8.1 Hz, 1 H), 6.76 (dm, J = 7.8 Hz, 1 H), 6.69 (m, 1 H), 6.46 (d, J = 1.5 Hz, 1 H), 4.98 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.22 (s, 2H), 2.11 (m, 4H), 1.43 (m, 4H), 1.36 (d, J = 7.0 Hz, 3H), 1.35 (m, 2H) tR: 3.12 min; MS(ESI): m/z (M+H)+557; (M+H)-555 Compound 26 (67 mg, 47%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1S)-1-(4-fluorophenyl)ethanamine C36H35FN4O2, Mw calc. : 574.3 1H-NMR in DMSO-d6 δ: 12.02 (s, 1 H), 10.93 (s, 1 H), 8.14 (d, J = 7.8 Hz, 1 H), 7.49-7.57 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.31 (ddm, J = 8.6 and 5.6 Hz, 2H), 7.12 (ddm, J = 9.0 and 8.6 Hz, 2H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.93 (dm, J = 7.6 Hz, 1 H), 6.88 (d, J = 8.1 Hz, 1 H), 6.76 (dm, J = 7.8 Hz, 1 H), 6.69 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 6.46 (d, J = 1.5 Hz, 1 H), 4.98 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.20 (s, 2H), 2.10 (m, 4H), 1.42 (m, 4H), 1.35 (d, J = 7.0 Hz, 3H), 1.34 (m, 2H) tR: 3.17 min; MS(ESI): m/z (M+H)+575; (M+H)-573 Compound 27 (67 mg, 47%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-(4-methylphenyl)ethanamine C37H38N4O2, Mw calc.: 570.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.04 (d, J = 7.8 Hz, 1 H), 7.49-7.55 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.16 (dm, J = 8.0 Hz, 2H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 7.10 (dm, J = 8.0 Hz, 2H), 6.92 (dm, J = 7.6 Hz, 1 H), 6.87 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.68 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 6.46 (d, J ∼ 1.0 Hz, 1 H), 4.94 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.21 (s, 2H), 2.26 (s, 3H), 2.10 (m, 4H), 1.42 (m, 4H), 1.35 (m, 2H), 1.34 (d, J = 7.0 Hz, 3H) tR: 3.16 min; MS(ESI): m/z (M+H)+571; (M+H)-569 Compound 28 (58 mg, 38%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1S)-1-(4-methylphenyl)ethanamine C37H38N4O2, Mw calc.: 570.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.04 (d, J = 7.8 Hz, 1H), 7.49-7.55 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.16 (dm, J = 8.0 Hz, 2H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 7.10 (dm, J = 8.0 Hz, 2H), 6.92 (dm, J = 7.6 Hz, 1 H), 6.87 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.68 (dd, J ∼ 2.5 and 2.0 Hz, 1 H), 6.46 (d, J ∼ 1.0 Hz, 1 H), 4.94 (dq, J = 7.8 and 7.0 Hz, 1 H), 3.21 (s, 2H), 2.26 (s, 3H), 2.10 (m, 4H), 1.42 (m, 4H), 1.35 (m, 2H), 1.34 (d, J = 7.0 Hz, 3H) tR: 3.26 min; MS(ESI): m/z (M+H)+571; (M+H)-569 Compound 29 (72 mg, 49%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[3-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-(4-methoxyphenyl)ethanamine C37H38N4O3, Mw calc.. 586.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.02 (d, J = 8.0 Hz, 1 H), 7.49-7.55 (ovl. m, 3H), 7.48 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.20 (dm, J = 8.6 Hz, 2H), 7.12 (dd, J = 7.8 and 7.6 Hz, 1 H), 6.93 (dm, J = 7.6 Hz, 1 H), 6.87 (d, J = 8.1 Hz, 1 H), 6.86 (dm, J = 8.6 Hz, 2H), 6.75 (dm, J = 7.8 Hz, 1 H), 6.68 (dd, J ∼ 2.5 and 2.0 Hz, 1H), 6.45 (d, J ∼ 1.0 Hz, 1H), 4.94 (dq, J = 8.0 and 7.0 Hz, 1H), 3.72 (s, 3H), 3.20 (s, 2H), 2.10 (m, 4H), 1.42 (m, 4H), 1.35 (m, 2H), 1.34 (d, J = 7.0 Hz, 3H) tR: 3.02 min; MS(ESI): m/z (M+H)+587; (M+H)-585 Compound 30 (60 mg, 43%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1S)-1-phenylethanamine C36H36N4O2, Mw calc.: 556.3 1H-NMR in DMSO-d6 δ: 12.02 (s, 1 H), 10.91 (s, 1 H), 8.10 (d, J = 8.0 Hz, 1 H), 7.50-7.56 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.20 8m, 1H), 7.04 (dm, J = 8.3 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 8.3 Hz, 2H), 6.45 (d, J = 1.0 Hz, 1 H), 4.98 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.29 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.36 d, J = 7.0 Hz, 3H)1.35 (m, 2H), tR: 3.16 min; MS(ESI): m/z (M+H)+557; (M+H)-555 Compound 31 (66 mg, 47%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-phenylethanamine C36H36N4O2, Mw calc.: 556.3 1H-NMR in DMSO-d6 δ: 12.02 (s, 1 H), 10.91 (s, 1 H), 8.10 (d, J = 8.0 Hz, 1 H), 7.50-7.56 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.20 8m, 1H), 7.04 (dm, J = 8.3 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 8.3 Hz, 2H), 6.45 (d, J = 1.0 Hz, 1 H), 4.98 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.29 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.36 d, J = 7.0 Hz, 3H)1.35 (m, 2H), tR: 3.16 min; MS(ESI): m/z (M+H)+557; (M+H)-555 Compound 32 (65 mg, 45%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1S)-1-phenylpropan-1-amine C37H38N4O2, Mw calc.: 570.3 1H-NMR in DMSO-d6 δ: 12.04 (s, 1 H), 10.91 (s, 1 H), 8.03 (d, J = 8.3 Hz, 1 H), 7.49-7.57 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.21 (m, 1 H), 7.04 (dm, J = 8.2 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.75 (dm, J = 8.2 Hz, 2H), 6.46 (d, J = 1.5 Hz, 1 H), 4.73 (ddd, J = 8.3, 7.3 and 7.3 Hz, 1 H), 3.29 (s, 2H), 2.23 (m, 4H), 1.60-1.80 (m, 2H), 1.44 (m, 4H), 1.36 (m, 2H), 0.81 (t, J = 7.2 Hz, 3H) tR: 3.25 min; MS(ESI): m/z (M+H)+571; (M+H)-569 Compound 33 (70 mg, 48%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1S)-1-(4-methoxyphenyl)ethanamine C37H38N4O3, Mw calc.: 586.3 1H-NMR in DMSO-d6 δ: 12.02 (s, 1 H), 10.91 (s, 1 H), 8.02 (d, J = 8.0 Hz, 1 H), 7.50-7.57 (ovl. m, 3H), 7.48 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.45 (m, 2H), 7.19 (dm, J = 8.6 Hz, 2H), 7.04 (dm, J = 8.3 Hz, 2H), 6.87 (d, J = 8.1 Hz, 1 H), 6.86 (dm, J = 8.6 Hz, 2H), 6.75 (dm, J = 8.3 Hz, 2H), 6.44 (d, J = 1.5 Hz, 1 H), 4.94 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.72 (s, 3H), 3.29 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.36 (m, 2H), 1.34 d, J = 7.0 Hz, 3H) tR: 3.07 min; MS(ESI): m/z (M+H)+587; (M+H)-585 Compound 34 (70 mg, 48%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1S)-1-(3-methoxyphenyl)ethanamine C37H38N4O3, Mw calc.: 586.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.92 (s, 1 H), 8.08 (d, J = 8.0 Hz, 1 H), 7.50-7.55 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.21 (dd, J = 8.0 and 7.8 Hz, 1 H), 7.05 (dm, J = 8.3 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.87 (dd, J ∼ 1.5 and 1.5 Hz, 1 H), 6.85 (dm, J = 7.8 Hz, 1 H), 6.78 (dm, J = 8.0 Hz, 1 H); 6.75 (dm, J = 8.3 Hz, 2H), 6.44 (d, J = 1.0 Hz, 1 H), 4.95 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.73 (s, 3H), 3.28 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.36 (m, 2H), 1.35 d, J = 7.0 Hz, 3H) tR: 3.09 min; MS(ESI): m/z (M+H)+587; (M+H)-585 Compound 35 (84 mg, 56%) was obtained starting from (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(phenyl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-(3-methoxyphenyl)ethanamine C37H39N5O3, Mw calc.: 601.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.10 (d, J = 8.0 Hz, 1 H), 7.50-7.56 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.21 (dd, J = 8.1 and 7.8 Hz, 1 H), 7.05 (dm, J = 8.3 Hz, 2H), 6.88 (d, J ∼ 8.1 Hz, 1 H), 6.87 (dd, J ∼ 2.0 and 1.5 Hz, 1 H), 6.85 (dm, J = 7.8 Hz, 1 H), 6.78 (dm, J = 8.1 Hz, 1 H), 6.76 (dm, J = 8.3 Hz, 2H), 6.44 (d, J = 1.0 Hz, 1 H), 4.95 (dq, J = 8.0 and 7.0 Hz, 1 H), 3.73 (s, 3H), 3.32 (s, 2H), 2.22-2.42 (m, 8H), 2.18 (s, 3H), 1.35 (d, J = 7.0 Hz, 3H) tR: 2.8 min; MS(ESI): m/z (M+H)+602; (M+H)-600 Compound 36 (72 mg, 47%) was obtained starting from (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(phenyl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-(3-chlorophenyl)ethanamine C36H36ClN5O2, Mw calc.: 605.2 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.18 (d, J = 7.7 Hz, 1 H), 7.50-7.57 (ovl. m, 3H), 7.50 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.35 (dd, J ∼ 1.5 and 1.5 Hz, 1 H), 7.33 (dd, J ∼ 7.5 and 7.5 Hz, 1 H), 7.27 (dm, J ∼ 7.5 Hz, 1 H), 7.24 (dm, J ∼ 7.5 Hz, 1 H), 7.05 (dm, J = 8.3 Hz, 2H), 6.89 (d, J ∼ 8.1 Hz, 1 H), 6.76 (dm, J = 8.3 Hz, 2H), 6.44 (d, J = 1.0 Hz, 1 H), 4.96 (dq, J = 7.7 and 7.0 Hz, 1 H), 3.31 (s, 2H), 2.19-2.39 (m, 8H), 2.14 (s, 3H), 1.36 (d, J = 7.0 Hz, 3H) tR: 3.00 min; MS(ESI): m/z (M+H)+606; (M+H)-604 Compound 37 (57 mg, 39%) was obtained starting from (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(phenyl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine C37H39N5O2, MW calc.: 585.3 1H-NMR in DMSO-d6 δ: 12.04 (s, 1 H), 10.93 (s, 1 H), 8.06 (d, J = 8.2 Hz, 1 H), 7.50-7.57 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.20 (m, 1 H), 7.05 (dm, J = 8.3 Hz, 2H), 6.88 (d, J ∼ 8.1 Hz, 1 H), 6.75 (dm, J = 8.3 Hz, 2H), 6.46 (d, J = 1.0 Hz, 1 H), 4.73 (ddd, J = 8.2, 7.4 and 7.4 Hz, 1 H), 3.29 (s, 2H), 2.21-2.41 (m, 8H), 2.16 (s, 3H), 1.60-1.80 (ovl. m, 2H), 0.81 (t, J = 7.0 Hz, 3H) tR: 2.91 min; MS(ESI): m/z (M+H)+586; (M+H)-584 Compound 38 (60 mg, 42%) was obtained starting from (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(phenyl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylethanamine C36H37N5O2, Mw calc.: 571.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.12 (d, J = 8.1 Hz, 1 H), 7.50-7.56 (ovl. m, 3H), 7.49 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.24-7.34 (ovl. m, 4H), 7.21 (m, 1 H), 7.06 (dm, J = 8.3 Hz, 2H), 6.88 (d, J ∼ 8.1 Hz, 1 H), 6.76 (dm, J = 8.3 Hz, 2H), 6.45 (d, J = 1.0 Hz, 1 H), 4.98 (dq, J = 8.1 and 7.0 Hz, 1 H), 3.32 (s, 2H), 2.23-2.43 (m, 8H), 2.18 (s, 3H), 1.36 (d, J = 7.0 Hz, 3H) tR: 2.78 min; MS(ESI): m/z (M+H)+572; (M+H)-570 Compound 39 (74 mg, 48%) was obtained starting from (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(phenyl)methylene]-2-oxoindoline-5-carboxylic acid and 1-(3,4-dimethoxyphenyl)methanamine C37H39N5O4, Mw calc.: 617.3 1H-NMR in DMSO-d6 δ: 12.04 (s, 1 H), 10.92 (s, 1 H), 8.32 (t, J = 5.5 Hz, 1 H), 7.49-7.57 (ovl. m, 3H), 7.46 (m, 2H), 7.45 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.06 (dm, J = 8.0 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.87 (d, J ∼ 8.0 Hz, 1 H), 6.86 (d, J ∼ 1.5 Hz, 1 H), 6.76 (dm, J = 8.0 Hz, 2H), 6.73 (dd, J ∼ 8.0 and 1.5 Hz, 1 H), 6.51 (d, J = 1.0 Hz, 1 H), 4.25 (d, J = 5.5 Hz, 2H), 3.72 (s, 3H), 3.71 (s, 3H), 3.36 (s, 2H), 2.31-2.33 (m, 8H), 2.30 (s, 3H) tR: 2.50 min; MS(ESI): m/z (M+H)+618; (M+H)-616 Compound 40 (72 mg, 49%) was obtained starting from (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(phenyl)methylene]-2-oxoindoline-5-carboxylic acid and 1-(3-methoxyphenyl)methanamine C36H37N5O3, Mw calc.: 587.3 1H-NMR in DMSO-d6 δ: 12.03 (s, 1 H), 10.93 (s, 1 H), 8.40 (t, J = 5.6 Hz, 1 H), 7.49-7.55 (ovl. m, 3H), 7.47 (dd, J = 8.1 and 1.0 Hz, 1 H), 7.46 (m, 2H), 7.21 (dd, J = 8.4 and 8.0 Hz, 1 H), 7.05 (dm, J = 8.3 Hz, 2H), 6.88 (d, J = 8.1 Hz, 1 H), 6.76-6.80 (ovl. m, 3H), 6.75 (dm, J = 8.3 Hz, 2H), 6.52 (d, J = 1.0 Hz, 1 H), 4.29 (d, J = 5.6 Hz, 2H), 3.72 (s, 3H), 3.28 (s, 2H), 2.19-2.33 (m, 8H), 2.12 (s, 3H) tR: 2.68 min; MS(ESI): m/z (M+H)+588; (M+H)-586 Compound 41 (72 mg, 49%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-6-carboxylic acid and (1 R)-1-phenylethanamine C36H36N4O2, Mw calc.: 556.3 1H-NMR in DMSO-d6 δ: 12.11 (s, 1 H), 10.85 (s, 1 H), 8.50 (d, J = 8.0 Hz, 1 H), 7.52-7.62 (ovl. m, 3H), 7.48 (m, 2H), 7.37 (d, J ∼ 1.5 Hz, 1 H), 7.34 (dm, J ∼ 7.5 Hz, 2H), 7.29 (ddm, J ∼ 7.5 and 7.0 Hz, 2H), 7.19 (tm, J ∼ 7.0 Hz, 1 H), 7.15 (dd, J = 8.1 and 1.5 Hz, 1 H), 7.06 (dm, J = 8.2 Hz, 2H), 6.80 (dm, J = 8.2 Hz, 2H), 5.71 (d, J = 8.1 Hz, 1 H), 5.11 (dq, J = 8.0, and 7.0 Hz, 1 H), 3.29 (s, 2H), 2.23 (m, 4H), 1.44 (m, 4H), 1.42 (d, J = 7.0 Hz, 3H)1.36 (m, 2H) tR: 3.15 min.; MS(ESI): m/z (M+H)+557; (M+H)-555 Compound 42 (77 mg, 54%) was obtained starting from (3Z)-2-oxo-3-(phenyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)indoline-6-carboxylic acid and (1R)-1-phenylpropan-1-amine C37H38N4O2, Mw calc.: 570.3 1H-NMR in DMSO-d6 δ: 12.10 (s, 1 H), 10.85 (s, 1 H), 8.43 (d, J = 8.4 Hz, 1 H), 7.52-7.62 (ovl. m, 3H), 7.48 (m, 2H), 7.35 (d, J ∼ 1.5 Hz, 1 H), 7.34 (dm, J ∼ 7.5 Hz, 2H), 7.28 (ddm, J ∼ 7.5 and 7.0 Hz, 2H), 7.19 (tm, J ∼ 7.0 Hz, 1H), 7.14 (dd, J = 8.2 and 1.5 Hz, 1 H), 7.06 (dm, J = 8.3 Hz, 2H), 6.80 (dm, J = 8.3 Hz, 2H), 5.70 (d, J = 8.1 Hz, 1 H), 4.84 (ddd, J = 8.0, 7.5 and 7.5 Hz, 1 H), 3.28 (s, 2H), 2.22 (m, 4H), 1.66-1.86 (ovl. m, 2H), 1.44 (m, 4H), 1.36 (m, 2H)0.85 (t, J = 7.3 Hz, 3H) tR: 3.24 min ; MS(ESI): m/z (M+H)+571; (M+H)-569 C36H37N5O2, Mw calc.:571.3 MS(ESI): m/z (M+H)+572 C37H39N5O2, Mw calc.:585.3 MS(ESI): m/z (M+H)+586 C35H35N5O2, Mw calc.:557.3 MS(ESI): m/z (M+H)+558 C36H36ClN5O2, Mw calc.:605.2 MS(ESI): m/z (M+H)+ 606 C37H39N5O3, Mw calc. :601.3 MS(ESI): m/z (M+H)+602 C36H37N5O3, Mw calc.:587.3 MS(ESI): m/z (M+H)+588 C37H39N5O4, Mw calc.:617.3 MS(ESI): m/z (M+H)+618 10 mmol (1.91 g) methyl-2-oxoindoline-5-carboxylate was suspended in 20 ml acetic-anhydride and stirred for 2-3 hours at 130-140 °C. The reaction mixture was cooled to rt and poured into crushed ice (100-150 g) then the precipitated product was filtered off. 2.17 g (93 %) Rt: 3.21 min. MS (ESI) [M+H]+: 234 MS (ESI) [M+H]-: 232 10 mmol (methyl-1-acetyl-2-oxoindoline-5-carboxylate) was dissolved in 40 ml dry dichloromethane and 100 mmol DIPEA, 0.1 mmol DMAP (4-Dimethylaminopyridine) were added to the reaction mixture under inert atmosphere and stirred for 5-10 mins. 1.1 mmol isonicotinoyl chloride was added in one portion and stirred for 2 hours. The reaction was monitored by TLC. After the completion, the reaction mixture was washed with 20-30 ml sat. aq. NaHCO3 and the organic layer was separated and dried over MgSO4. The dessicant was filtered off and the solvent was removed under reduced pressure to give methyl-(3Z)-1-acetyl-3-[hydroxy(pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylate as brown foamy solid. C18H14N2O5 calc. 338.32 MS (ESI) [M+H]+: 339 MS (ESI) [M+H]-: 337 The following compounds were obtained according to the procedure described by example 2, step B
10 mmol (3.38 g) methyl-(3Z)-1-acetyl-3-[hydroxy(pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylate intermediate was suspended in 40 ml dry dioxane and 20 mmol (3.22 g, 4.18 ml) HMDS (hexamethyldisilazane), 20 mmol (2.17 g, 2.54 ml) trimethylsilyl chloride and 4-[(4-methylpiperazin-1-yl)methyl]aniline were added to the reaction mixture under inert atmosphere and refluxed overnight. The reaction mixture was poured into 30 ml 10 % Na2CO3 and extracted with 3x60 ml ethyl-acetate. The organic layer was separated and dried over MgSO4. The dessicant was filtered off and the solvent was removed under reduced pressure to give methyl-(3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylate as dark brown oil. This product was used in the next step without purification. C28H29N5O3 calc. 483.58 MS (ESI) [M+H]+: 484 MS (ESI) [M+H]-: 482 The following compounds were obtained according to the procedure described by example 2, step C
10 mmol (4.83g) Methyl-(3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino) (pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylate was suspended in mixture of 50 ml metanol and 10 ml water and 20 mmol (0.80 g) NaOH was added and stirred at 60 °C for 4-6 hours. The reaction was monitored by TLC. After the completion the pH was adjusted to pH = 4-5 and the precipitated product was filtered off. C27H27N5O3 calc. 469.55 MS (ESI) [M+H]+: 470 MS (ESI) [M+H]-: 468 The following compounds were obtained according to the procedure described by example 2, step D
0.5 mmol (230 mg) (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino) (pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylic acid, 0.5 mmol (68 mg) (1R)-1-phenylpropan-1-amine, 0.6 mmol (0.450 g) HATU/HBTU , 1.5 mmol (0.19 g, 0.26 ml)DIPEA were stirred in 8 ml dry DMF at 50 °C for 2-5 hours. The solvent was removed under reduced pressure and the residue was taken up with 20-30 ml sat. K2CO3 and extracted with 3x30 ml ethyl-acetate. The organic layer was separated and and dried over MgSO4. The dessicant was filtered off and the solvent was removed under reduced pressure. The product was purified by column chromatography (Kieselgel, chloroform/methanol (sat. with NH315:1) C36H38N6O2 calc.: 586.74 MS (ESI) [M+H]+: 587 MS (ESI) [M+H]- : 585 The following compounds were obtained according to the procedure described by example 2, step E Compound 136 was obtained by reaction of (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and benzyl-amine according to Step E. C34H34N6O2 calc.: 558.69 MS (ESI) [M+H]+: 559 MS (ESI) [M+H]- : 557 C34H33FN6O2 calc.: 576.68 MS (ESI) [M+H]+: 577 MS (ESI) [M+H]- : 575 Compound 137 was obtained by reaction of (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and benzyl-amine according to Step E. Compound 138 was obtained by reaction of (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl} amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylethanamine according to Step E. C35H36N6O2 calc.: 572.72 MS (ESI) [M+H]+: 573 MS (ESI) [M+H]- : 571 Compound 139 was obtained by reaction of (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl} amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C36H38N6O2 calc.: 586.74 MS (ESI) [M+H]+: 587 MS (ESI) [M+H]- : 585 Compound 140 was obtained by reaction (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1R)-1-phenylethanamine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 141 was obtained by reaction (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1S)-1-phenylethanamine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 142 was obtained by reaction (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 143 was obtained by reaction (3Z)-3-[({4-[(diethylamino)methyl]phenyl}amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H37N5O2 calc.: 559.72 MS (ESI) [M+H]+: 560 MS (ESI) [M+H]- : 558 Compound 144 was obtained by reaction (3Z)-3-[({4-[(diethylamino)methyl]phenyl} amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylethanamine according to Step E. C34H35N5O2 calc.: 545.69 MS (ESI) [M+H]+: 546 MS (ESI) [M+H]- : 544 Compound 145 was obtained by reaction (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1S)-1-phenylpropan-1-amine according to Step E. C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 146 was obtained by reaction and (3Z)-3-[{[4-(morpholin-4-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylethanamine according to Step E. C34H33N5O3 calc.: 559.67 MS (ESI) [M+H]+: 560 MS (ESI) [M+H]- : 558 Compound 147 was obtained by reaction of (3Z)-3-[{[4-(morpholin-4-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H35N5O3 calc.: 573.70 MS (ESI) [M+H]+: 574 MS (ESI) [M+H]- : 572 Compound 148 was obtained by reaction of (3Z)-3-[({3-[(4-methylpiperazin-1-yl)methyl]phenyl} amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C35H36N6O2 calc.: 572.72 MS (ESI) [M+H]+: 573 MS (ESI) [M+H]- : 571 Compound 149 was obtained by reaction of (3Z)-3-[({3-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(pyridin-3-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C36H38N6O2 calc.: 586.74 MS (ESI) [M+H]+: 587 MS (ESI) [M+H]- : 585 Compound 150 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 151 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1S)-1-phenylpropan-1-amine according to Step E. C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 152 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1S)-1-phenylethanamine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 153 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-3-yl)methylene]indoline-5-carboxylic acid and (1R)-1-phenylethanamine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 154 was obtained by reaction of and and according to (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-2-yl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 155 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-2-yl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. Compound 156 was obtained by reaction of (3Z)-2-oxo-3-(pyridin-2-yl{[3-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H33N5O2 calc.: 543.67 MS (ESI) [M+H]+: 544 MS (ESI) [M+H]- : 542 Compound 157 was obtained by reaction of (3Z)-2-oxo-3-(pyridin-2-yl{[3-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 158 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-2-yl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 159 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-2-yl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 160 was obtained by reaction of and (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C35H36N6O2 calc.: 572.72 MS (ESI) [M+H]+: 573 MS (ESI) [M+H]- : 571 Compound 161 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-4-yl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 162 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino} (pyridin-4-yl)methylene]indoline-5-carboxylic acid and (1S)-1-phenylpropan-1-amine according to Step E. C36H37N5O2 calc.: 571.73 MS (ESI) [M+H]+: 572 MS (ESI) [M+H]- : 570 Compound 163 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-4-yl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 164 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino}(pyridin-4-yl)methylene]indoline-5-carboxylic acid and (1S)-1-phenylpropan-1-amine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 165 was obtained by reaction of (3Z)-3-[({4-[(diethylamino)methyl]phenyl}amino) (pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H35N5O2 calc.: 545.69 MS (ESI) [M+H]+: 546 MS (ESI) [M+H]- : 544 Compound 166 was obtained by reaction of (3Z)-3-[({4-[(diethylamino)methyl]phenyl}amino)(pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H37N5O2 calc. 559.72 MS (ESI) [M+H]+: 560 MS (ESI) [M+H]- : 558 Compound 167 was obtained by reaction of (3Z)-3-[{[4-(morpholin-4-ylmethyl)phenyl]amino}(pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H33N5O3 calc.: 559.67 MS (ESI) [M+H]+: 560 MS (ESI) [M+H]- : 558 Compound 168 was obtained by reaction of (3Z)-3-[{[4-(morpholin-4-ylmethyl)phenyl]amino}(pyridin-4-yl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H35N5O3 calc.: 573.70 MS (ESI) [M+H]+: 574 MS (ESI) [M+H]- :572 Compound 169 was obtained by reaction of (3Z)-2-oxo-3-(pyridin-4-yl{[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C35H35N5O2 calc.: 557.70 MS (ESI) [M+H]+: 558 MS (ESI) [M+H]- : 556 Compound 170 was obtained by reaction of (3Z)-2-oxo-3-(pyridin-4-yl{[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene)indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H33N5O2 calc.: 543.67 MS (ESI) [M+H]+: 544 MS (ESI) [M+H]- : 542 Compound 171 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(pyrrolidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C33H32N4O2S calc.: 548.71 MS (ESI) [M+H]+: 549 MS (ESI) [M+H]- : 547 Compound 172 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(pyrrolidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C34H34N4O2S calc.:562.74 MS (ESI) [M+H]+: 563 MS (ESI) [M+H]- : 561 Compound 173 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H34N4O2S calc.: 562.74 MS (ESI) [M+H]+: 563 MS (ESI) [M+H]- : 561 Compound 174 was obtained by reaction of (3Z)-2-oxo-3-[{[4-(piperidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and(1R)-1-phenylpropan-1-amine according to Step E. C35H36N4O2S calc.: 576.77 MS (ESI) [M+H]+: 577 MS (ESI) [M+H]- : 575 Compound 175 was obtained by reaction of (3Z)-3-[{[4-(morpholin-4-ylmethyl)phenyl]amino}(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C33H32N4O3S calc.: 564.71 MS (ESI) [M+H]+: 565 MS (ESI) [M+H]- : 563 Compound 176 was obtained by reaction of (3Z)-3-[{[4-(morpholin-4-ylmethyl)phenyl]amino}(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C34H34N4O3S calc.: 578.74 MS (ESI) [M+H]+: 579 MS (ESI) [M+H]- : 577 Compound 177 was obtained by reaction of (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H35N5O2S calc.: 577.75 MS (ESI) [M+H]+: 578 MS (ESI) [M+H]- : 576 Compound 178 was obtained by reaction of (3Z)-3-[({4-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H37N5O2S calc.: 591.78 MS (ESI) [M+H]+: 592 MS (ESI) [M+H]- : 590 Compound 179 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(pyrrolidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C33H32N4O2S calc.: 548.71 MS (ESI) [M+H]+: 549 MS (ESI) [M+H]- : 547 Compound 180 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(pyrrolidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C34H34N4O2S calc.: 562.74 MS (ESI) [M+H]+: 563 MS (ESI) [M+H]- : 561 Compound 181 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H34N4O2S calc.: 562.74 MS (ESI) [M+H]+: 563 MS (ESI) [M+H]- : 561 Compound 182 was obtained by reaction of (3Z)-2-oxo-3-[{[3-(piperidin-1-ylmethyl)phenyl]amino}(3-thienyl)methylene]indoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C35H36N4O2S calc.: 576.77 MS (ESI) [M+H]+: 577 MS (ESI) [M+H]- : 575 Compound 183 was obtained by reaction of (3Z)-3-[({3-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H35N5O2S calc.: 577.75 MS (ESI) [M+H]+: 578 MS (ESI) [M+H]- : 576 Compound 184 was obtained by reaction of (3Z)-3-[({3-[(4-methylpiperazin-1-yl)methyl]phenyl}amino)(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H37N5O2S calc.: 591.78 MS (ESI) [M+H]+: 592 MS (ESI) [M+H]- : 590 Compound 185 was obtained by reaction of (3Z)-3-[{[3-(morpholin-4-ylmethyl)phenyl]amino}(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C33H32N4O3S calc.: 564.71 MS (ESI) [M+H]+: 565 MS (ESI) [M+H]- : 563 Compound 186 was obtained by reaction of (3Z)-3-[{[3-(morpholin-4-ylmethyl)phenyl]amino}(3-thienyl)methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C34H34N4O3S calc.: 578.74 MS (ESI) [M+H]+: 579 MS (ESI) [M+H]- : 577 Compound 187 was obtained by reaction of (3Z)-3-(3-furyl{[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H34N4O3 calc.: 532.65 MS (ESI) [M+H]+: 533 MS (ESI) [M+H]-: 531 Compound 188 was obtained by reaction of (3Z)-3-(3-furyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C35H36N4O3 calc.: 560.70 MS (ESI) [M+H]+: 561 MS (ESI) [M+H]- : 559 Compound 189 was obtained by reaction of (3Z)-3-(2-furyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and according to (1R)-1-phenylpropan-1-amine preparation of according to, Step E. C35H36N4O3 calc.: 560.70 MS (ESI) [M+H]+: 561 MS (ESI) [M+H]- : 559 Compound 190 was obtained by reaction of and according to preparation of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[4-(pyrrolidin-1-ylmethyl)phenyl]aminolmethylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylethanamine, according to Step E. C33H34N6O2 calc.: 546.68 MS (ESI) [M+H]+: 547 MS (ESI) [M+H]- : 545 Compound 191 was obtained by reaction of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C34H36N6O2 calc.: 560.71 MS (ESI) [M+H]+: 560 MS (ESI) [M+H]- : 558 Compound 192 was obtained by reaction of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[4-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C34H36N6O2 calc.: 560.71 MS (ESI) [M+H]+: 561 MS (ESI) [M+H]- : 559 Compound 193 was obtained by reaction of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[4-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H38N6O2 calc.: 574.73 MS (ESI) [M+H]+: 575 MS (ESI) [M+H]- : 573 Compound 194 was obtained by reaction of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[3-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C33H34N6O2 calc.: 546.68 MS (ESI) [M+H]+: 547 MS (ESI) [M+H]- : 545 Compound 195 was obtained by reaction of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[3-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C34H36N6O2 calc.: 560.71 MS (ESI) [M+H]+: 561 MS (ESI) [M+H]- : 559 Compound 196 was obtained by reaction of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[3-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid (1 R)-1-phenylethanamine and according to Step E. C34H36N6O2 calc.: 560.71 MS (ESI) [M+H]+: 561 MS (ESI) [M+H]- : 559 Compound 197 was obtained by reaction of (3Z)-3-[(1-methyl-1H-pyrazol-4-yl){[3-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C35H38N6O2 calc.: 574.73 MS (ESI) [M+H]+: 575 MS (ESI) [M+H]- : 573 Compound 198 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C38H37N5O2 calc.: 595.75 MS (ESI) [M+H]+: 596 MS (ESI) [M+H]- : 594 Compound 199 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C39H39N5O2 calc.: 609.78 MS (ESI) [M+H]+: 610 MS (ESI) [M+H]- : 608 Compound 200 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[4-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C39H39N5O2 calc.: 609.78 MS (ESI) [M+H]+: 610 MS (ESI) [M+H]- : 608 Compound 201 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[4-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C40H41N5O2 calc.: 623.81 MS (ESI) [M+H]+: 624 MS (ESI) [M+H]- : 622 Compound 202 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[3-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C38H37N5O2 calc.: 595.75 MS (ESI) [M+H]+: 596 [M+H]- : 594 Compound 203 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[3-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1R)-1-phenylpropan-1-amine according to Step E. C39H39N5O2 calc.: 609.78 MS (ESI) [M+H]+: 610 [M+H]- : 608 Compound 204 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[3-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C39H39N5O2 calc.: 609.78 MS (ESI) [M+H]+: 610 [M+H]- : 608 Compound 205 was obtained by reaction of (3Z)-3-[(1-methyl-1H-indol-5-yl){[3-(piperidin-1-ylmethyl)phenyl]amino}methylene]-2-oxoindoline-5-carboxylic acid (1R)-1-phenylpropan-1-amine according to Step E. C40H41N5O2 calc.: 623.81 MS (ESI) [M+H]+: 624 [M+H]- : 622 Compound 206 was obtained by reaction of (3Z)-3-(cyclopropyl{[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C32H34N4O2 calc.: 506.65 MS (ESI) [M+H]+: 507 [M+H]- : 505 Compound 207 was obtained by reaction of (3Z)-3-(cyclopropyl{[4-(pyrrolidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C33H36N4O2 calc.: 520.68 MS (ESI) [M+H]+: 521 [M+H]- : 519 Compound 208 was obtained by reaction of (3Z)-3-(cyclopropyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C33H36N4O2 calc.: 520.68 MS (ESI) [M+H]+: 521 [M+H]- : 519 Compound 209 was obtained by reaction of (3Z)-3-(cyclopropyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C34H38N4O2 calc.: 534.71 MS (ESI) [M+H]+: 535 [M+H]- : 533 Compound 210 was obtained by reaction of (3Z)-3-(cyclohexyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C36H42N4O2 calc.: 562.76 MS (ESI) [M+H]+: 563 [M+H]- : 561 Compound 211 was obtained by reaction of (3Z)-3-(cyclohexyl{[4-(piperidin-1-ylmethyl)phenyl]amino}methylene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C37H44N4O2 calc.: 576.79 MS (ESI) [M+H]+: 577 [M+H]- : 575 Compound 212 was obtained by reaction of (3Z)-2-oxo-3-(1-{[4-(piperidin-1-ylmethyl)phenyl]amino}propylidene)indoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C32H36N4O2 calc. 508.67 MS (ESI) [M+H]+ 509 [M+H]- :507 Compound 213 was obtained by reaction of (3Z)-2-oxo-3-(1-{[4-(piperidin-1-ylmethyl)phenyl]amino}propylidene)indoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C33H38N4O2, calc: 522.70 MS (ESI) [M+H]+: 523 [M+H]- : 521 Compound 214 was obtained by reaction of (3Z)-3-(2-methyl-1-{[4-(piperidin-1-ylmethyl)phenyl]amino}propylidene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylethanamine according to Step E. C33H38N4O2, calc. 522.70 MS (ESI) [M+H]+: 523 [M+H]- : 521 Compound 215 was obtained by reaction of (3Z)-3-(2-methyl-1-{[4-(piperidin-1-ylmethyl)phenyl]amino}propylidene)-2-oxoindoline-5-carboxylic acid and (1 R)-1-phenylpropan-1-amine according to Step E. C34H40N4O2, calc. 536.72 MS (ESI) [M+H]+: 537 [M+H]- : 535 C36H35FN4O2, Mw calc. :574.3 MS(ESI): m/z (M+H)+575 C37H37FN4O2, Mw cal. :588.3 MS(ESI): m/z (M+H)+589 C37H37FN4O3, Mw calc. :604.3 MS(ESI): m/z (M+H)+605 C37H37FN4O3, Mw calc.:604.3 MS(ESI): m/z (M+H)+605 C36H34ClFN4O2, Mw calc. :608.2 MS(ESI): m/z (M+H)+609 C36H34ClFN4O2, Mw calc. :608.2 MS(ESI): m/z (M+H)+609 C37H37FN4O2, Mw calc. :588.3 MS(ESI): m/z (M+H)+589 C37H37FN4O2, Mw calc. : 624.3 MS(ESI): m/z (M+H)+625 C37H36ClFN4O2, Mw calc. :622.2 MS(ESI): m/z (M+H)+623 MS (ESI) [M+H]+: 547 MS (ESI) [M+H]+: 549 MS (ESI) [M+H]+: 629 MS (ESI) [M+H]+: 625 MS (ESI) [M+H]+: 625 MS (ESI) [M+H]+: 559 MS (ESI) [M+H]+: 648 MS (ESI) [M+H]+: 561 MS (ESI) [M+H]+: 643 MS (ESI) [M+H]+: 563 MS (ESI) [M+H]+: 625 MS (ESI) [M+H]+: 693 MS (ESI) [M+H]+: 782 MS (ESI) [M+H]+: 577 MS (ESI) [M+H]+: 643 MS (ESI) [M+H]+: 625 MS (ESI) [M+H]+: 560 MS (ESI) [M+H]+: 648 MS (ESI) [M+H]+: 583 MS (ESI) [M+H]+: 667 MS (ESI) [M+H]+: 562 MS (ESI) [M+H]+: 644 MS (ESI) [M+H]+: 574 MS (ESI) [M+H]+: 663 MS (ESI) [M+H]+: 645 MS (ESI) [M+H]+: 559 MS (ESI) [M+H]+: 573 MS (ESI) [M+H]+: 662 MS (ESI) [M+H]+: 648 MS (ESI) [M+H]+: 589 MS (ESI) [M+H]+: 672 MS (ESI) [M+H]+: 575 MS (ESI) [M+H]+: 575 MS (ESI) [M+H]+: 657 MS (ESI) [M+H]+: 662 MS (ESI) [M+H]+: 575 MS (ESI) [M+H]+:657 MS (ESI) [M+H]+: 536 MS (ESI) [M+H]+: 536 MS (ESI) [M+H]+:522 MS (ESI) [M+H]+: 494 MS (ESI) [M+H]+: 494 MS (ESI) [M+H]+: 508 MS (ESI) [M+H]+: 509 MS (ESI) [M+H]+: 486 The activity of the compounds described in the present invention was determined by the following kinase assays. The inhibition of Akt1 by the compounds described in the present invention were determined measuring the phosphorylation of a fluorescently-labeled peptide (TAMRA-peptide) by human Akt1 kinase by fluorescent polarization using a commercially available IMAP Screening Express Assay Kit (Molecular Devices). Akt1 kinase assays were performed in low protein binding 384-well plates (Corning). Test compounds were diluted in 100% DMSO to 5 mM stock concentration and then further dilutions were made in H2O or 100% DMSO to desirable concentrations. Each reaction consisted of 5 nM enzyme: Akt1 kinase, 400 nM TAMRA-peptide (5-TAMRA-GRTGRRNSI-COOH; Genecust), 40 µM ATP and kinase buffer: 20 mM MES pH6, 1 mM DTT, 10 mM MgCl2, 2 mM MnCl2, 0.01 V/V% TritonX (all reagents from Sigma-Aldrich). For each reaction, 4 or 6 µl containing TAMRA-peptide, ATP and kinase buffer were combined with 2 µl diluted compound in H2O or 0.02 µl compound in 100% DMSO. The kinase reaction was started by the addition of 2 µl diluted enzyme. The reaction was allowed to run for 1 hour at room temperature. The reaction was stopped by adding 15 µl IMAP beads (1:400 beads in progressive (75% buffer A, 25% buffer B) 1x buffer). After an additional hour, fluorescent polarization (Ex: 550-10 nm, Em: 590-10 nm, Dich: 561 nm) was measured using an Analyst GT (Molecular Devices). The activity of the compounds described in the present invention can be determined by the following kinase assay, which measures the generation of ADP by human Ack1 kinase by fluorescent polarization using a commercially available TranScreener ADP2 FP Assay Kit (BellBrook Labs). Ack1 kinase assays were performed in low protein binding 384-well plates (Corning). Test compounds were diluted in 100% DMSO to 5 mM stock concentration and then further dilutions were made in H2O or 100% DMSO to desirable concentrations. Each reaction consisted of 5 nM enzyme: Ack1 kinase, Poly Glu-Tyr (4:1) (Sigma), 3.5 µM ATP and kinase buffer: 20 mM HEPES pH7.5, 1 mM DTT, 2 mM MgCl2, 0.4 mM MnCl2, 0.01 V/V% Tween20 (all reagents from Sigma-Aldrich) For each reaction, 4 or 6 µl containing Poly Glu-Tyr (4:1), ATP and kinase buffer were combined with 2 µl diluted compound in H2O or 0.02 µl compound in 100% DMSO. The kinase reaction was started by the addition of 2 µl diluted enzyme. The reaction was allowed to run for 1 hour at room temperature. The reaction was stopped by addition of 8 µl ADP detection mixture (1 x) (1 x Detection buffer + 4.78 µg/ml ADP2 Ab + 4 nM ADP Alexa633 Tracer). After an additional hour, fluorescent polarization (Ex: 630-35 nm, Em: 680-30 nm, Dich: 650 nm) was measured using an Analyst GT (Molecular Devices). The activity of the compounds described in the present invention can be determined by the following kinase assay, which measures the phosphorylation of a fluorescently-labeled peptide by human Clk1 kinase by fluorescent polarization using a commercially available IMAP Screening Express Assay Kit (Molecular Devices). Clk1 kinase assays were performed in low protein binding 384-well plates (Corning). Test compounds were diluted in 100% DMSO to 5 mM stock concentration and then further dilutions were made in H2O or 100% DMSO to desirable concentrations. Each reaction consisted of 4 nM enzyme: Clk1 kinase domain, 400 nM TAMRA-AFRREWSPGKEAKK-NH2 (Genecust), 0.45 µM ATP (Sigma) and kinase buffer: 20 mM TRIS pH 8, 1 mM DTT, 0.4 mM MgCl2, 0.4 mM MnCl2, 0.01 V/V% Tween20 (all reagents from Sigma-Aldrich) For each reaction, 4 or 6 µl containing TAMRA-peptide, ATP and kinase buffer were combined with 2 µl diluted compound in H2O or 0.02 µl compound in 100% DMSO. The kinase reaction was started by the addition of 2 µl diluted enzyme. The reaction was allowed to run for 1 hour at room temperature. The reaction was stopped by adding 15 µl IMAP beads (1:600 beads in progressive (80 % buffer A, 20% buffer B) 1x buffer). After an additional hour, fluorescent polarization (Ex: 550-10 nm, Em: 590-10 nm, Dich: 561 nm) was measured using an Analyst GT (Molecular Devices). Human lung epithelial A549 cells were pre-treated on 384 well plates with the various substances in eight different 1:3 dilutions two hours before infection. Infection was performed using 130 plaque forming units (PFU) of the H1 N1 influenza virus strain A/WSN/33. Subsequently, compounds were re-added at the same concentrations. 36 hours post infection virus-containing supernatants were transferred to Madin-Darby Canine Kidney Epithelial (MDCK) cells, seeded the day before, to quantify the virus titer. The MDCK cells were fixed with formaldehyde 7 hours post infection and the percentage of infected to non-infected cells was quantified by staining cells with an influenza virus specific antibody (recognizing the viral nucleoprotein) and with Hoechst dye (to label all cell nuclei), followed by automated microscopy and single object analysis. Based on the results of the eight different dilutions dose-response-curves could be calculated and the IC50 values of each compound were determined. All experiments were performed within the BSL-3 lab using the FXP Laboratory Automation Workstation (Beckman Coulter). The whole screening process is illustrated in To be able to exclude possible cytotoxic effects all compounds were additionally tested using the 'Cell Proliferation Reagent WST-1' (Roche Diagnostics). Therefore, A549 cells were treated with the same concentrations of the various substances and 36 hours later the WST-1 was added to the cells. Finally, the read-out of this test was conducted at 48 hours post treatment using the EnVision Multilabel Reader (PerkinElmer). A549 human lung carcinoma cells were cultured in RPMI 1640 medium (Sigma) supplemented with 10% fetal bovine serum (Sigma) and Antibiotic-Antimycotic (Sigma) in tissue culture flasks at 37°C in a humidified 5% CO2 atmosphere. Cells were routinely assayed for mycoplasma contamination. Two thousand cells were plated in 384-well tissue culture plates, and treated after 24 hours with a 10-point dilution series of the compounds (1.5 nM-30 uM), using the vehicle, DMSO as control. Cells were incubated for 24 hours after which the relative cell density was determined by CellTiter-Glo Luminescent Cell Viability Assay (Promega). This method determines the number of metabolically active cells in the culture based on quantitation of the ATP present. The luminescent signal was measured with an Analyst GT multimode reader (Molecular Devices). Samples were diluted in DMSO (control), phosphate buffer pH = 7.4 and pH = 2.0, from 5mM stock solutions (solved in DMSO), with a 120 µM final concentration. The samples were incubated for 24 hours at room temperature followed by 30 minutes centrifugation at 3700 rpm. 40 µl of the supernatants were injected into HPLC (Waters 2795 Alliance HPLC equipped with Waters 996 PDA Detector (HPLC column: Waters XBridge, RP C18, 3.5 µm, 4.6 mm x 50 mm; gradient MeCN/H2O, containing 0.1% HCOOH: 5% MeCN (0.5 min), 5% to 95% MeCN (5 min), 95% MeCN (0.5 min); flow rate: 2.0 ml/min)) and the AUC values (measured on sample specific wavelength) of the buffered samples were divided by the AUC values (same wavelength as buffered samples') of the DMSO control samples. HCT-116 and HKE3 human colon carcinoma cell lines (two completely identical cell lines that differ only in KRas mutation status) and HepG2 hepatoblastomaderived cell line were cultured in RPMI 1640 medium (Sigma) supplemented with 10% fetal bovine serum (Sigma) and Antibiotic-Antimycotic (Sigma) in tissue culture flasks at 37°C in a humidified 5% CO2 atmosphere. Cell lines were routinely assayed for mycoplasma contamination. One thousand cells were plated in 384-well tissue culture plates, and treated after 24 hours with a 10-point dilution series of the compounds (1.5 nM-30 µM), using the vehicle, DMSO as control. Cells were incubated for 72 hours after which the relative cell density was determined by CellTiter-Glo Luminescent Cell Viability Assay (Promega). This method determines the number of metabolically active cells in the culture based on quantitation of the ATP present. The luminescent signal was measured with an Analyst GT multimode reader (Molecular Devices). |
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