Patent Application
20100227863
The present invention relates to novel substituted 3-(4-hydroxyphenyl)-indolin-2-one compounds (oxindole compounds), and the use of such compounds for the preparation of a medicament for the treatment of cancer in a mammal.
U.S. Pat. No. 1,624,675 describes O—O-diacyl derivatives of diphenolisatine and that these compounds possess laxative properties.
US 2004/0242563 A 1 discloses substituted diphenyl indanone, indane and indole compounds and analogues thereof useful for the treatment or prevention of diseases characterized by abnormal cell proliferation.
Magnus et al. (Magnus P and Turnbull R (2006) Organic Letters 8(16): 3497-3499) describe the synthesis of the following oxindole:
Felding et al. (WO 2005/097107) describe a number of oxindoles as anti-cancer agents, e.g. the following oxindoles:
Halperin et al. (WO 2005/080335) describe a number of oxindoles as potential anti-cancer agents, e.g. the following oxindoles;
Baskakova et al. (SU 90-4875262) describe the following oxindole for the manufacture of optical articles.
Kawada et al. (JP 94-114510) describe the following oxindole as a resist agent:
Hosta Pujol et al. (DE 2521966) describe the synthesis of the following oxindole as a potential laxative:
Esteve Subirana et al. (DE 2451592) describe the following oxindole as a laxative agent:
Kornowski (Kornowski H (1963) Bulletin de la Societe Chimique de France 10: 2035-2036) describes the synthesis of the following oxindole:
Aktiebolaget “Ferrosan” ((1957) British patent application no. GB 1955-34509) describes the following oxindole as a laxative:
Geigy J R ((1955) British patent application no. GB 1952-23426) describes the synthesis of the following oxindoles:
Luk et al. (WO 2006/136606) describe oxindoles as potential anticancer agents:
Notably, these compounds comprise only one R1 substituent. Also neither R2 nor R4 are para-hydroxyphenyl.
Although, Felding et al. and Halperin et al. describe various ox-indole-2-one-type compounds as anti-cancer agents, there is still a need for novel ox-indol-2-one-type compounds as anti-cancer agents which provide useful alternatives upon selection of drug candidates.
The present inventors have now found that a new class of compounds represents an excellent alternative to existing ox-indol-2-one-type compounds as anti-cancer agents, and that the new compounds have comparative or even improved potency compared to the known compounds.
Hence, the present invention provides compounds of the general formulae (I) and (Ia), cf. claims 1, 24 and 25.
The present invention further provides a pharmaceutical composition, cf. claim 26, the utilization of compounds of the general formulae (I) and (Ia) in medicine, cf. claims 28, 29 and 31.
The present invention i.a. relates to particular prodrug compounds which are useful for the treatment of cancer in a mammal.
The useful prodrug compounds have the general formula (I), namely
wherein
r is 0 or 1;
X is selected from —CH2—, —O—, —S—, —S(O)—, —S(O)2— and —NR5—, wherein R5 is selected from hydrogen and optionally substituted C1-6-alkyl;
Z is selected from optionally substituted C1-12-alkyl, optionally substituted C3-12-cycloalkyl, optionally substituted C2-12-alkenyl, optionally substituted C3-12-cycloalkenyl, optionally substituted C2-12-alkynyl, optionally substituted heterocyclyl, optionally substituted aryl and optionally substituted heteroaryl;
with the proviso that Z is not para-mono-substituted phenyl when r is 0, in particular not mono-substituted phenyl;
V1, V2, V3, and V4 independently are selected from a carbon atom, a non-quaternary nitrogen atom, an oxygen atom, and a sulphur atom, and where V4 further may be selected from a bond, so that -V1-V2-V3-V4- together with the atoms to which V1 and V4 are attached form an aromatic or heteroaromatic ring;
R1, R2, R3, and R4, when attached to a carbon atom, independently are selected from hydrogen, optionally substituted C1-12-alkyl, optionally substituted C3-12-cycloalkyl, optionally substituted C2-12-alkenyl, optionally substituted C3-12-cycloalkenyl, hydroxy, optionally substituted C1-12-alkoxy, optionally substituted C2-12-alkenyloxy, carboxy, optionally substituted C1-12-alkoxycarbonyl, optionally substituted C1-12-alkylcarbonyl, optionally substituted C1-12-alkylcarbonyloxy, formyl, amino, mono- and di(C1-12-alkyl)amino, carbamoyl, mono- and di(C1-12-alkyl)aminocarbonyl, C1-12-alkylcarbonylamino, C1-12-alkylsulphonylamino, cyano, carbamido, mono- and di(C1-12-alkyl)amino-carbonylamino, C1-12-alkanoyloxy, C1-12-alkylsulphonyl, C1-12-alkylsulphinyl, aminosulphonyl, mono- and di(C1-12-alkyl)aminosulphonyl, nitro, optionally substituted C1-12-alkylthio, aryl, aryloxy, arylcarbonyl, arylamino, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylcarbonyl, heteroaryl, heteroaryloxy, heteroarylamino, heteroarylcarbonyl, and halogen, where any C1-12-alkyl as an amino substituent is optionally substituted with hydroxy, C1-12-alkoxy, amino, mono- and di(C1-12-alkyl)amino, carboxy, C1-12-alkylcarbonylamino, C1-12-alkylaminocarbonyl, or halogen(s), and wherein any aryl, heterocyclyl and heteroaryl may be optionally substituted;
R1, R2, R3, and R4, when attached to a nitrogen atom, independently are selected from hydrogen, optionally substituted C1-12-alkyl, hydroxy, oxide, optionally substituted C1-12-alkoxy, optionally substituted C1-12-alkoxycarbonyl, optionally substituted C1-12-alkylcarbonyl, formyl, mono- and di(C1-12-alkyl)aminocarbonyl, amino, C1-12-alkylcarbonylamino, mono- and di(C1-12-alkyl)amino, C1-12-alkylsulphonyl, C1-12-alkylsulphinyl, aryl, aryloxy, arylcarbonyl, arylamino, heterocyclyl, heterocyclyloxy, heterocyclylcarbonyl, heterocyclylamino, heteroaryl, heteroaryloxy, heteroarylcarbonyl, and heteroarylamino; where any C1-12-alkyl as an amino substituent is optionally substituted with hydroxy, C1-12-alkoxy, amino, mono- and di(C1-12-alkyl)amino, carboxy, C1-12-alkylcarbonylamino, C1-12-alkylaminocarbonyl, or halogen(s), and wherein any aryl, heterocyclyl and heteroaryl may be optionally substituted;
or R1 and R2 together with the carbon atoms to which they are attached form a ring;
with the proviso that at least one of the substituents R1, R2, R3, and R4 is not hydrogen;
and pharmaceutically acceptable salts and prodrugs thereof.
In the present context, the terms “C1-12-alkyl” and “C1-6-alkyl” are intended to mean a linear, cyclic or branched hydrocarbon group having 1 to 12 carbon atoms and 1 to 6 carbon atoms, respectively, such as methyl, ethyl, propyl, iso-propyl, pentyl, cyclopentyl, hexyl, cyclohexyl. The term “C1-4-alkyl” is intended to cover linear, cyclic or branched hydrocarbon groups having 1 to 4 carbon atoms, e.g. methyl, ethyl, propyl, iso-propyl, cyclopropyl, butyl, iso-butyl, tert-butyl, cyclobutyl.
Although the term “C3-12-cycloalkyl” is encompassed by the term “C1-12-alkyl”, it refers specifically to the mono- and bicyclic counterparts, including alkyl groups having exo-cyclic atoms, e.g. cyclohexyl-methyl.
Similarly, the terms “C2-12-alkenyl” and “C2-6-alkenyl” are intended to cover linear, cyclic or branched hydrocarbon groups having 2 to 12 carbon atoms and 2 to 6 carbon atoms, respectively, and comprising (at least) one unsaturated bond. Examples of alkenyl groups are vinyl, allyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, heptadecaenyl. Preferred examples of alkenyl are vinyl, allyl, butenyl, especially allyl.
Although the term “C3-12-cycloalkenyl” is encompassed by the term “C2-12-alkenyl”, it refers specifically to the mono- and bicyclic counterparts, including alkenyl groups having exo-cyclic atoms, e.g. cyclohexenyl-methyl.
In the present context, i.e. in connection with the terms “alkyl”, “cycloalkyl”, “alkylidene”, “alkoxy”, “alkenyl”, “cycloalkenyl” and the like, the term “optionally substituted” is intended to mean that the group in question may be substituted one or several times, preferably 1-3 times, with group(s) selected from hydroxy (which when bound to an unsaturated carbon atom may be present in the tautomeric keto form), C1-6-alkoxy (i.e. C1-6-alkyl-oxy), C2-6-alkenyloxy, carboxy, oxo (forming a keto or aldehyde functionality), C1-6-alkoxycarbonyl, C1-6-alkylcarbonyl, formyl, aryl, aryloxy, arylamino, arylcarbonyl, aryloxycarbonyl, arylcarbonyloxy, arylaminocarbonyl, arylcarbonylamino, heteroaryl, heteroaryloxy, heteroarylamino, heteroarylcarbonyl, heteroaryloxycarbonyl, heteroarylcarbonyloxy, heteroarylaminocarbonyl, heteroarylcarbonylamino, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylcarbonyl, heterocyclyloxycarbonyl, heterocyclylcarbonyloxy, heterocyclylaminocarbonyl, heterocyclylcarbonylamino, amino, mono- and di(C1-6-alkyl)amino, —N(C1-4-alkyl)3+, carbamoyl, mono- and di(C1-6-alkyl)aminocarbonyl, C1-6-alkyl-carbonylamino, cyano, guanidino, carbamido, C1-6-alkyl-sulphonyl-amino, aryl-sulphonyl-amino, heteroaryl-sulphonyl-amino, C1-6-alkanoyloxy, C1-6-alkyl-sulphonyl, C1-6-alkyl-sulphinyl, C1-6-alkylsulphonyloxy, nitro, C1-6-alkylthio, and halogen, where any aryl, heteroaryl and heterocyclyl may be substituted as specifically described below for aryl, heteroaryl and heterocyclyl, and any alkyl, alkoxy, and the like, representing substituents may be substituted with hydroxy, C1-6-alkoxy, amino, mono- and di(C1-6-alkyl)amino, carboxy, C1-6-alkylcarbonylamino, C1-6-alkylaminocarbonyl, or halogen(s).
Typically, the substituents are selected from hydroxy (which when bound to an unsaturated carbon atom may be present in the tautomeric keto form), C1-6-alkoxy (i.e. C1-6-alkyl-oxy), C2-6-alkenyloxy, carboxy, oxo (forming a keto or aldehyde functionality), C1-6-alkylcarbonyl, formyl, aryl, aryloxy, arylamino, arylcarbonyl, heteroaryl, heteroaryloxy, heteroarylamino, heteroarylcarbonyl, heterocyclyl, heterocyclyloxy, heterocyclylamino, heterocyclylcarbonyl, amino, mono- and di(C1-6-alkyl)amino; carbamoyl, mono- and di(C1-6-alkyl)aminocarbonyl, amino-C1-6-alkyl-aminocarbonyl, mono- and di(C1-6-alkyl)amino-C1-6-alkyl-aminocarbonyl, C1-6-alkylcarbonylamino, guanidino, carbamido, C1-6-alkyl-sulphonyl-amino, C1-6-alkyl-sulphonyl, C1-6-alkyl-sulphinyl, C1-6-alkylthio, halogen, where any aryl, heteroaryl and heterocyclyl may be substituted as specifically described below for aryl, heteroaryl and heterocyclyl.
In some embodiments, substituents are selected from hydroxy, C1-6-alkoxy, amino, mono- and di(C1-6-alkyl)amino, carboxy, C1-6-alkylcarbonylamino, C1-6-alkylaminocarbonyl, or halogen.
The term “halogen” includes fluoro, chloro, bromo, and iodo.
In the present context, the term “aryl” is intended to mean a fully or partially aromatic carbocyclic ring or ring system, such as phenyl, naphthyl, 1,2,3,4-tetrahydronaphthyl, anthracyl, phenanthracyl, pyrenyl, benzopyrenyl, fluorenyl and xanthenyl, among which phenyl is a preferred example.
The term “heteroaryl” is intended to mean a fully or partially aromatic carbocyclic ring or ring system where one or more of the carbon atoms have been replaced with heteroatoms, e.g. nitrogen (═N— or —NH—), sulphur, and/or oxygen atoms. Examples of such heteroaryl groups are oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, coumaryl, furanyl, thienyl, quinolyl, benzothiazolyl, benzotriazolyl, benzodiazolyl, benzooxozolyl, phthalazinyl, phthalanyl, triazolyl, tetrazolyl, isoquinolyl, acridinyl, carbazolyl, dibenzazepinyl, indolyl, benzopyrazolyl, phenoxazonyl. Particularly interesting heteroaryl groups are benzimidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, furyl, thienyl, quinolyl, triazolyl, tetrazolyl, isoquinolyl, indolyl in particular benzimidazolyl, pyrrolyl, imidazolyl, pyridinyl, pyrimidinyl, furyl, thienyl, quinolyl, tetrazolyl, and isoquinolyl.
The term “heterocyclyl” is intended to mean a non-aromatic carbocyclic ring or ring system where one or more of the carbon atoms have been replaced with heteroatoms, e.g. nitrogen (═N— or —NH—), sulphur, and/or oxygen atoms. Examples of such heterocyclyl groups (named according to the rings) are imidazolidine, piperazine, hexahydropyridazine, hexahydro-pyrimidine, diazepane, diazocane, pyrrolidine, piperidine, azepane, azocane, aziridine, azirine, azetidine, pyrroline, tropane, oxazinane (morpholine), azepine, dihydroazepine, tetrahydroazepine, and hexahydroazepine, oxazolane, oxazepane, oxazocane, thiazolane, thiazinane, thiazepane, thiazocane, oxazetane, diazetane, thiazetane, tetrahydrofuran, tetrahydropyran, oxepane, tetrahydrothiophene, tetrahydrothiopyrane, thiepane, dithiane, dithiepane, dioxane, dioxepane, oxathiane, oxathiepane. The most interesting examples are tetrahydrofuran, imidazolidine, piperazine, hexahydropyridazine, hexahydropyrimidine, diazepane, diazocane, pyrrolidine, piperidine, azepane, azocane, azetidine, tropane, oxazinane (morpholine), oxazolane, oxazepane, thiazolane, thiazinane, and thiazepane, in particular tetrahydrofuran, imidazolidine, piperazine, hexahydropyridazine, hexahydropyrimidine, diazepane, pyrrolidine, piperidine, azepane, oxazinane (morpholine), and thiazinane.
In the present context, i.e. in connection with the terms “aryl”, “benzylidene”, “heteroaryl”, “heterocyclyl” and the like (e.g. “aryloxy”, “heterarylcarbonyl”, etc.), the term “optionally substituted” is intended to mean that the group in question may be substituted one or several times, preferably 1-5 times, in particular 1-3 times, with group(s) selected from hydroxy (which when present in an enol system may be represented in the tautomeric keto form), C1-6-alkyl, C1-6-alkoxy, C2-6-alkenyloxy, oxo (which may be represented in the tautomeric enol form), oxide (only relevant as the N-oxide), carboxy, C1-6-alkoxycarbonyl, C1-6-alkylcarbonyl, formyl, aryl, aryloxy, arylamino, aryloxycarbonyl, arylcarbonyl, heteroaryl, heteroarylamino, amino, mono- and di(C1-6-alkyl)amino; carbamoyl, mono- and di(C1-6-alkyl)aminocarbonyl, amino-C1-6-alkyl-aminocarbonyl, mono- and di(C1-6-alkyl)amino-C1-6-alkyl-aminocarbonyl, C1-6-alkylcarbonylamino, cyano, guanidino, carbamido, C1-6-alkanoyl-oxy, C1-6-alkyl-sulphonyl-amino, aryl-sulphonyl-amino, heteroaryl-sulphonyl-amino, C1-6-alkyl-suphonyl, C1-6-alkyl-sulphinyl, C1-6-alkylsulphonyloxy, nitro, sulphanyl, amino, amino-sulphonyl, mono- and di(C1-6-alkyl)amino-sulphonyl, dihalogen-C1-4-alkyl, trihalogen-C1-4-alkyl, halogen, where aryl and heteroaryl representing substituents may be substituted 1-3 times with C1-4-alkyl, C1-4-alkoxy, nitro, cyano, amino or halogen, and any alkyl, alkoxy, and the like, representing substituents may be substituted with hydroxy, C1-6-alkoxy, C2-6-alkenyloxy, amino, mono- and di(C1-6-alkyl)amino, carboxy, C1-6-alkylcarbonylamino, halogen, C1-6-alkylthio, C1-6-alkyl-sulphonyl-amino, or guanidino.
Typically, the substituents are selected from hydroxy, C1-6-alkyl, C1-6-alkoxy, oxo (which may be represented in the tautomeric enol form), carboxy, C1-6-alkylcarbonyl, formyl, amino, mono- and di(C1-6-alkyl)amino; carbamoyl, mono- and di(C1-6-alkyl)aminocarbonyl, amino-C1-6-alkyl-aminocarbonyl, C1-6-alkylcarbonylamino, guanidino, carbamido, C1-6-alkyl-sulphonyl-amino, aryl-sulphonyl-amino, heteroaryl-sulphonyl-amino, C1-6-alkyl-suphonyl, C1-6-alkyl-sulphinyl, C1-6-alkylsulphonyloxy, sulphanyl, amino, amino-sulphonyl, mono- and di(C1-6-alkyl)amino-sulphonyl or halogen, where any alkyl, alkoxy and the like, representing substituents may be substituted with hydroxy, C1-6-alkoxy, C2-6-alkenyloxy, amino, mono- and di(C1-6-alkyl)amino, carboxy, C1-6-alkylcarbonylamino, halogen, C1-6-alkylthio, C1-6-alkyl-sulphonyl-amino, or guanidino. In some embodiments, the substituents are selected from C1-6-alkyl, C1-6-alkoxy, amino, mono- and di(C1-6-alkyl)amino, sulphanyl, carboxy or halogen, where any alkyl, alkoxy and the like, representing substituents may be substituted with hydroxy, C1-6-alkoxy, C2-6-alkenyloxy, amino, mono- and di(C1-6-alkyl)amino, carboxy, C1-6-alkylcarbonylamino, halogen, C1-6-alkylthio, C1-6-alkyl-sulphonyl-amino, or guanidino.
The term “prodrug” used herein is intended to mean a compound which—upon exposure to physiological conditions—will liberate a derivative said compound which then will be able to exhibit the desired biological action.
The term “pharmaceutically acceptable salts” is intended to include acid addition salts and basic salts. Illustrative examples of acid addition salts are pharmaceutically acceptable salts formed with non-toxic acids. Exemplary of such organic salts are those with maleic, fumaric, benzoic, ascorbic, succinic, oxalic, bis-methylenesalicylic, methanesulphonic, ethanedisulphonic, acetic, propionic, tartaric, salicylic, citric, gluconic, lactic, malic, mandelic, cinnamic, citraconic, aspartic, stearic, palmitic, itaconic, glycolic, p-aminobenzoic, glutamic, benzenesulphonic, and theophylline acetic acids, as well as the 8-halotheophyllines, for example 8-bromotheophylline. Exemplary of such inorganic salts are those with hydrochloric, hydrobromic, sulphuric, sulphamic, phosphoric, and nitric acids. Examples of basic salts are salts where the (remaining) counter ion is selected from alkali metals, such as sodium and potassium, alkaline earth metals, such as calcium, and ammonium ions (+N(R)3R′, where R and R′ independently designates optionally substituted C1-6-alkyl, optionally substituted C2-6-alkenyl, optionally substituted aryl, or optionally substituted heteroaryl). Pharmaceutically acceptable salts are, e.g., those described in Remington's Pharmaceutical Sciences, 17. Ed. Alfonso R. Gennaro (Ed.), Mack Publishing Company, Easton, Pa., U.S.A., 1985 and more recent editions and in Encyclopedia of Pharmaceutical Technology. Thus, the term “an acid addition salt or a basic salt thereof” used herein is intended to comprise such salts. Furthermore, the compounds as well as any intermediates or starting materials may also be present in hydrate form.
Moreover, it should be understood that the compounds may be present as racemic mixtures or the individual stereoisomers such as enantiomers or diastereomers. The present invention encompasses each and every of such possible stereoisomers (e.g. enantiomers and diastereomers) as well as racemates and mixtures enriched with respect to one of the possible stereoisomers.
It should be understood that relevant feature of the compounds of the formula (I) include that the group Z is not para-mono-substituted phenyl (in particular not mono-substituted) when r is 0, and at least one of the substituents R1, R2, R3, and R4 is not hydrogen. Preferably, at least two of the substituents R1, R2, R3, and R4 are not hydrogen;
It appears that the group Z (as defined hereinabove) plays an important role for the optimization of the biological activity of the compounds.
This being said, Z is in one interesting embodiment selected from optionally substituted C1-12-alkyl, optionally substituted C3-12-cycloalkyl, optionally substituted C2-12-alkenyl, optionally substituted C3-12-cycloalkenyl, optionally substituted C2-12-alkynyl, and optionally substituted heterocyclyl.
In one variant hereof, Z is selected from C1-12-alkyl, C3-12-cycloalkyl, C2-12-alkenyl, C3-12-cycloalkenyl, and C2-12-alkynyl.
In another variant hereof, Z is selected from optionally substituted C3-12-cycloalkyl and optionally substituted heterocyclyl (e.g. piperidine and morpholine), in particular from C3-12-cycloalkyl, heterocyclyl, and mono-substituted heterocyclyl.
In another interesting embodiment, Z is optionally substituted heteroaryl, in particular heteroaryl.
In a still further interesting embodiment, Z is aryl or, alternatively, Z is di- or tri-substituted aryl.
The orientation of the group Z is also in part defined by the presence (r=1) and type of the group X.
In one interesting embodiment, r is 1 and X is —CH2—.
In another interesting embodiment, r is 0.
The atoms V1, V2, V3, and V4 define whether the ring is an aromatic or heteroaromatic ring. Besides an aromatic ring (a benzene ring), a plethora of aromatic rings are possible.
In one particularly interesting embodiment, however, each of V1, V2, V3, and V4 represents a carbon atom (a benzene ring), or V3 represents a nitrogen atom and each of V1, V2, and V4 represents a carbon atom (a pyridine ring). In the currently most interesting embodiments, each of V1, V2, V3, and V4 represents a carbon atom (i.e. the ring is a benzene ring).
The substituents R1 and R2 of the substituents R1, R2, R3, and R4 seem to play a particular role.
Preferably, R1 is selected from halogen, C1-6-alkyl, trifluoromethyl and C1-6-alkoxy, when V1 is a carbon atom.
Also preferably, R2 is selected from halogen, optionally substituted C1-6-alkyl, and optionally substituted C1-6-alkoxy, when V2 is a carbon atom.
Further, it is preferred that R3 is selected from hydrogen, optionally substituted C1-6-alkoxy, halogen, cyano, optionally substituted aryl, optionally substituted aryloxy, optionally substituted heteroaryl, amino, C1-6-alkylcarbonylamino, C1-6-alkylsulphonylamino, and mono- and di(C1-6-alkyl)aminosulphonyl, when V3 is a carbon atom.
Even further, it is preferred that R4 is hydrogen, when V4 is a carbon atom.
This being said, it is preferred that at least two of the substituents R1, R2, R3, and R4 are not hydrogen.
In one variant hereof, R3 and R4 are both hydrogen.
In a further variant hereof, none of R1 and R2 are hydrogen. In a particular variant, R1 and R2 are both selected from halogen and methyl. In a specific variant hereof, R1 and R2 are both fluoro.
Alternatively, R1 and R2 together with the carbon atoms to which they are attached form a ring selected from aromatic rings, carbocyclic rings, heterocyclic rings and heteroaromatic rings, in particular aromatic rings, heterocyclic rings and heteroaromatic rings.
It has been found that certain compounds wherein R3 and R4 are both hydrogen and wherein none of R1 and R2 are hydrogen represent a particularly interesting aspect of the present invention. Hence, the present invention also provides a compound of the general formula (Ia)
wherein Z, R1 and R2 are as defined herein, with the proviso that none of R1 and R2 are hydrogen.
Presently very interesting compounds of the formulae (I) and (Ia) are those listed in the following:
The compounds of the present invention can be prepared in a number of ways well known to those skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the methods outlined below and in the Examples section, together with methods known in the art of organic synthetic organic chemistry, or variations thereof as appreciated by those skilled in the art. Preferred methods include, but are not limited to, those described below.
The novel compounds of formula (I) may be prepared using the reactions and techniques described in this section. The reactions are performed in solvents appropriate to the reagents and materials employed and suitable for the transformations being effected. Also, in the synthetic methods described below, it is to be understood that all proposed reaction conditions, including choice of solvent, reaction atmosphere, reaction temperature duration of experiment and work-up procedures, are chosen to be conditions of standard for that reaction, which should be readily recognized by one skilled in the art. It is understood by one skilled in the art of organic synthesis that the functionality present on various portions of the educt molecule must be compatible with the reagents and reactions proposed. Not all molecules of formula (I) falling into a given class may be compatible with some of the reaction conditions required in some of the methods described. Such restrictions to the substituents which are compatible with the reaction conditions will be readily apparent to one skilled in the art and alternative methods can be used.
Compounds of general formula (I), in which r is 0 or X is —CH2— can be prepared from an isatin-derivative by reaction with a Grignard-reagent or an organolithium reagent to form tertiary alcohols of general formula (II), which are subsequently allowed to react with phenol in a Friedel-Craft reaction in the presence of an acid, e.g. p-toluenesulphonic acid (p-TSA).
Isatin derivatives are either commercially available or can be prepared as described in the literature (e.g. Stolle: J. Prakt. Chem. (1922), 105, 137; Sandmeyer: Helv. Chim. Acta (1919), 2, 234; Shvedov et al.: Chem. Heterocycl. Comp. Engl. Transl. (1975). 11, 666; Hewawasam and Maenwell: Tet. Lett. (1994). 35, 7303; Rivalle and Bisagani: J. Heterocycl. Chem. (1997), 34, 441; Tatsugi et al.: ARKIVOC (2001), 67; Silva et al.: J. Braz. Chem. Soc. (2001), 12, 273).
Compounds (I) according to the present invention in which X is —CH2— can also be prepared from tertiary alcohols of general formula (II), in which r is 0 and Z is a protected p-hydroxyphenyl (IIa), in which Pg is a protecting group (e.g. methyl, t-butyl, benzyl, triisopropylsilyl or other silyl protecting groups, tetrahydropyranyl, acetyl, benzoyl etc.), by dehydroxylation to yield deoxygenated intermediates of general formula (III), which are subsequently treated with a base (e.g. n-butyllithium and N,N,N,N-tetramethylehylenediamine) and an alkylating agent such as an alkylhalide to yield compounds of general formula (IV), followed by deprotection to yield compounds of general formula (I).
Compounds (I) according to the present invention, in which X is —NR5—, —O— or —S— can be prepared from tertiary alcohols of general formula (IIa), by conversion of the alcohol into a leaving group such as the chloro-compounds of general formula (V) and subsequent reaction with an amine, alcohol or thiol in the presence of a base, such as for instance diisopropylethylamine or sodium hydride, to yield intermediates of general formula (VI), and subsequent removal of the protecting group.
Compounds (I) according to the present invention in which X is —S(O)— or —S(O)2— can be prepared from compounds of general formula (I) in which X is —S— by oxidation, e.g. by use of m-chloroperbenzoic acid in equimolar amount or excess, respectively.
Compounds (I) according to the present invention in which r is 0 and Z is imidazol attached via nitrogen can be prepared from tertiary alcohols (IIa) by reaction with 1,1′-carbonyldiimidazole to yield intermediates of general formula (VII) and subsequent removal of the protecting group.
Compounds (I) according to the present invention in which X═CH2— and Z is C1-12-alkylcarbonyl, arylcarbonyl and heteroarylcarbonyl can be prepared from isatin derivatives in a Knoevenagel condensation with the corresponding ketones to yield intermediates of general formula (IIb), which are subsequently allowed to react with phenol in a Friedel-Craft reaction in the presence of an acid, e.g. p-TSA.
Compounds (I) according to the present invention which are racemates, can be resolved into the enantiomers by purification on a chiral column, e.g. Daicel Chiralcel-OD.
The compounds of the general formulae (I) and (Ia) are believed to be particularly useful in the treatment of cancer. The term cancer is typically describing cell growth not under strict control. In one embodiment of the invention, treatment of cancers in which inhibition of protein synthesis and/or inhibition of activation of the mTOR pathway is an effective method for reducing cell growth. Examples of such cancers include, but are not limited to, breast cancer, renal cancer, multiple myeloma, leukemia, glioblastoma, rhabdomyosarcoma, prostate, soft tissue sarcoma, colorectal sarcoma, gastric carcinoma, head and neck squamous cell carcinoma, uterine, cervical, melanoma, lymphoma, and pancreatic cancer.
Any type of cell may be treated, including but not limited to, lung, gastrointestinal (including e.g. bowel, colon), breast (mammary), ovarian, prostate, liver (hepatic), kidney (renal), bladder, pancreas, brain and skin.
Hence, the present invention generally provides a compound of the general formula (I) or (Ia) as defined herein for use as a medicament; more particular, the use of a compound of the general formula (I) or (Ia) as defined herein for the preparation of a medicament for the treatment of cancer in a mammal. Such medicaments may further comprise one or more other chemotherapeutic agents.
Moreover, the present invention provides a method of treating a mammal suffering from or being susceptible to cancer, the method comprising administering to the mammal a therapeutically effective amount of a compound of the general formula (I) or (Ia) as defined herein.
The compounds of the general formulae (I) and (Ia) are suitably formulated in a pharmaceutical composition so as to suit the desirable route of administration.
The administration route of the compounds may be any suitable route which leads to a concentration in the blood or tissue corresponding to a therapeutic effective concentration. Thus, e.g., the following administration routes may be applicable although the invention is not limited thereto: the oral route, the parenteral route, the cutaneous route, the nasal route, the rectal route, the vaginal route and the ocular route. It should be clear to a person skilled in the art that the administration route is dependent on the particular compound in question; particularly the choice of administration route depends on the physico-chemical properties of the compound together with the age and weight of the patient and on the particular disease or condition and the severity of the same.
The compounds may be contained in any appropriate amount in a pharmaceutical composition, and are generally contained in an amount of about 1-95%, e.g. 1-10%, by weight of the total weight of the composition. The composition may be presented in a dosage form which is suitable for the oral, parenteral, rectal, cutaneous, nasal, vaginal and/or ocular administration route. Thus, the composition may be in form of, e.g., tablets, capsules, pills, powders, granulates, suspensions, emulsions, solutions, gels including hydrogels, pastes, ointments, creams, plasters, drenches, delivery devices, suppositories, enemas, injectables, implants, sprays, aerosols and in other suitable form.
The pharmaceutical compositions may be formulated according to conventional pharmaceutical practice, see, e.g., “Remington's Pharmaceutical Sciences” and “Encyclopedia of Pharmaceutical Technology”, edited by Swarbrick, J. & J. C. Boylan, Marcel Dekker, Inc., New York, 1988. Typically, the compounds defined herein are formulated with (at least) a pharmaceutically acceptable carrier or excipient. Pharmaceutically acceptable carriers or excipients are those known by the person skilled in the art. Formation of suitable salts of the compounds of the Formulae (I) and (Ia) will also be evident in view of the before-mentioned.
Thus, the present invention provides in a further aspect a pharmaceutical composition comprising a compound of the general Formula (I) or (Ia) in combination with a pharmaceutically acceptable carrier.
Pharmaceutical compositions according to the present invention may be formulated to release the active compound substantially immediately upon administration or at any substantially predetermined time or time period after administration. The latter type of compositions is generally known as controlled release formulations.
In the present context, the term “controlled release formulation” embraces i) formulations which create a substantially constant concentration of the drug within the body over an extended period of time, ii) formulations which after a predetermined lag time create a substantially constant concentration of the drug within the body over an extended period of time, iii) formulations which sustain drug action during a predetermined time period by maintaining a relatively, constant, effective drug level in the body with concomitant minimization of undesirable side effects associated with fluctuations in the plasma level of the active drug substance (saw-tooth kinetic pattern), iv) formulations which attempt to localize drug action by, e.g., spatial placement of a controlled release composition adjacent to or in the diseased tissue or organ, v) formulations which attempt to target drug action by using carriers or chemical derivatives to deliver the drug to a particular target cell type.
Controlled release formulations may also be denoted “sustained release”, “prolonged release”, “programmed release”, “time release”, “rate-controlled” and/or “targeted release” formulations.
Controlled release pharmaceutical compositions may be presented in any suitable dosage forms, especially in dosage forms intended for oral, parenteral, cutaneous nasal, rectal, vaginal and/or ocular administration. Examples include single or multiple unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, liposomes, delivery devices such as those intended for oral, parenteral, cutaneous, nasal, vaginal or ocular use.
Preparation of solid dosage forms for oral use, controlled release oral dosage forms, fluid liquid compositions, parenteral compositions, controlled release parenteral compositions, rectal compositions, nasal compositions, percutaneous and topical compositions, controlled release percutaneous and topical compositions, and compositions for administration to the eye will be well-known to those skilled in the art of pharmaceutical formulation. Specific formulations can be found in “Remington's Pharmaceutical Sciences”.
Capsules, tablets and pills etc. may contain for example the following compounds: microcrystalline cellulose, gum or gelatin as binders; starch or lactose as excipients; stearates as lubricants; various sweetening or flavouring agents. For capsules the dosage unit may contain a liquid carrier like fatty oils. Likewise coatings of sugar or enteric agents may be part of the dosage unit. The pharmaceutical compositions may also be emulsions of the compound(s) and a lipid forming a micellular emulsion.
For parenteral, subcutaneous, intradermal or topical administration the pharmaceutical composition may include a sterile diluent, buffers, regulators of tonicity and antibacterials. The active compound may be prepared with carriers that protect against degradation or immediate elimination from the body, including implants or microcapsules with controlled release properties. For intravenous administration the preferred carriers are physiological saline or phosphate buffered saline.
In one embodiment, the pharmaceutical composition is in unit dosage form. In such embodiments, each unit dosage form typically comprises 0.1-500 mg, such as 0.1-200 mg, e.g. 0.1-100 mg, of the compound.
More generally, the compound are preferably administered in an amount of about 0.1-250 mg per kg body weight per day, such as about 0.5-100 mg per kg body weight per day.
For compositions adapted for oral administration for systemic use, the dosage is normally 0.5 mg to 1 g per dose administered 1-4 times daily for 1 week to 12 months depending on the disease to be treated.
The dosage for oral administration of the composition in order to prevent diseases or conditions is normally 1 mg to 100 mg per kg body weight per day. The dosage may be administered once or twice daily for a period starting 1 week before the exposure to the disease until 4 weeks after the exposure.
For compositions adapted for rectal use for preventing diseases, a somewhat higher amount of the compound is usually preferred, i.e. from approximately 1 mg to 100 mg per kg body weight per day.
For parenteral administration, a dose of about 0.1 mg to about 100 mg per kg body weight per day is convenient. For intravenous administration, a dose of about 0.1 mg to about 20 mg per kg body weight per day administered for 1 day to 3 months is convenient. For intraarticular administration, a dose of about 0.1 mg to about 50 mg per kg body weight per day is usually preferable. For parenteral administration in general, a solution in an aqueous medium of 0.5-2% or more of the active ingredients may be employed.
For topical administration on the skin, a dose of about 1 mg to about 5 g administered 1-10 times daily for 1 week to 12 months is usually preferable.
In an intriguing embodiment of the present invention, the compound of the general formula (I) or (Ia) is used therapeutically in combination with one or more other chemotherapeutic agents. Examples of such chemotherapeutic agents are those selected from daunorubicin, docetaxel, prednisone, dexamethasone, decadron, altretamine, amifostine, aminoglutethimide, dactinomycin, anastrozole, asparaginase, bicalutamide, bleomycin, busulfan, carboplatin, carmustine, chlorambucil, chlorodeoxyadenosine, cisplatin, cytosine arabinoside, dacarbazine, doxorubicin, epirubicin, estramustine, diethylstilbestrol, fludarabine, flutamide, 5-fluorouracil, gemcitabine, goserelin, idarubicin, irinotecan, levamisole, lomustine, mechlorathamine, alkeran, mercaptopurine, taxol (e.g. paclitaxel). In particular, the further chemotherapeutic agent is selected from taxanes such as Taxol, Paclitaxel and Docetaxel.
Thus, with respect to the use and the method of treatment defined herein, the medicament may further comprise one or more other chemotherapeutic agents.
For nuclear magnetic resonance 1H-NMR spectra (300 MHz) and 13C-NMR (75.6) chemical shift values (δ) (in ppm) are quoted, unless otherwise specified, for deuterochloroform solutions relative to tetramethylsilane (δ=0.0) or chloroform (δ=7.25) or deuterochloroform (δ=76.81 for 13C-NMR) standards. The value of a multiplet, either defined (doublet (d), triplet (t) quartet (q)) or not (m) at the approximate mid point is given unless a range is quoted. (bs) indicates a broad singlet.
MS was performed using a Micromass LCT with an AP-ESI-probe or LC-MS using a Bruker Esquire 3000+ESI Iontrap with an Agilent 1200 HPLC-system.
The organic solvents used were anhydrous.
The following abbreviations have been used throughout:
DCM dichloromethane
DMAP N,N dimethylaminopyridine
EtOAc ethyl acetate
MS mass spectroscopy
NMR nuclear magnetic resonance
n-BuLi n-butyl lithium
rt room temperature
p-TSA para-toluenesulphonic acid
TFA trifluoroacetic acid
TLC thin layer chromatography
To a stirred solution of isatin derivative in dry THF under nitrogen at −78° C. was added 3 eq. of Grignard reagent or 3 eq. of freshly prepared solution of organolithium reagent. After 30 min, the dry-ice bath was removed and the reaction was left to reach room temperature over 4 to 14 hours. Excess Grignard reagent was quenched with water, and the reaction mixture was acidified with 1N HCl or saturated NH4Cl-solution, extracted with EtOAc (2×), dried over Mg2SO4, filtered and concentrated. The residue was purified by chromatography (1% methanol in DCM or mixtures of petroleum ether and EtOAc) to afford racemic compounds of general formula II).
To a solution of tertiary alcohol of general formula (II) in dichloroethane was added phenol (5 eq.) and p-TSA (7.5 eq.). The reaction mixture was heated to 90° C. for 2-4 hours and then cooled to room temperature. The solid (mainly p-TSA) was filtered off and washed with dichloroethane or DCM. The solution was concentrated and the residue was purified by chromatography (1% methanol in DCM or mixtures of petroleum ether and EtOAc) to afford racemic compounds of general formula (I).
A mixture of tertiary alcohol (IIa), Et3SiH (3 eq.) and TFA were heated to 100° C. in a sealed tube for 1-3 days until the deoxygenation was complete. Excess Et3SiH and TFA were evaporated, and the residue was purified by chromatography (1% methanol in DCM or mixtures of petroleum ether and EtOAc) to afford racemic compounds of general formula (III).
Compound of general formula (III) was dissolved in dry THF under nitrogen, TMEDA (2.2 eq.) was added and the mixture was cooled to −78° C. 1.6 M n-BuLi solution (2.2 eq.) was added dropwise and the mixture stirred at −78° C. for 0.5-1 hour. The alkylating agent (2.2 eq.) was then added and the reaction mixture gradually allowed to reach room temperature. After 3-8 hours the mixture was quenched with water, extracted with EtOAc (2×), dried over Mg2SO4, filtered and concentrated. The residue was purified by chromatography (1% methanol in DCM or mixtures of petroleum ether and EtOAc) to afford racemic compounds of general formula (IV).
Compound of general formula (IV), in which the protecting group is a methyl group, was dissolved in DCM under nitrogen, cooled to −78° C. and BBr3-solution (1.0 M, 1.5 eq.) was added dropwise with stirring. The reaction mixture was gradually allowed to reach room temperature. After 4-18 hours the mixture was quenched with water, extracted with Et2O (2×), dried over Mg2SO4, filtered and concentrated. The residue was purified by chromatography (1% methanol in DCM or mixtures of petroleum ether and EtOAc) to afford racemic compounds of general formula (I).
Racemic compound of general formula (I) was dissolved in ethanol or ethanol/heptane mixtures and purified by HPLC on Daicel Chiralcel-OD 250×20 mm ID 5 micron to yield pure enantiomers of general formula (I).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and ethynylmagnesium chloride. 1H-NMR (DMSO-d6) δ 11.33 (bs), 7.3-7.2 (m, 1H), 7.15 (s, 1H), 7.1-7.0 (m, 1H), 3.68 (s, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and 4-methoxymagnesium bromide. 1H-NMR (DMSO-d6) δ 11.12 (bs), 7.25-7.15 (d, 2H), 7.1-6.9 (m, 4H), 6.71 (s, 1H), 3.73 (s, 3H).
General procedure 3. Starting materials: Starting materials: 6,7-difluoro-3-hydroxy-3-(4-methoxyphenyl)indolin-2-one (compound 2). 1H-NMR (CDCl3) δ 8.08 (bs, 1H), 7.2-7.1 (d, 2H), 7.0-6.8 (m, 4H), 4.61 (s, 1H), 3.82 (s, 3H).
General procedure 4. Starting materials: Compound 3 and benzyl bromide. 1H-NMR (CDCl3) δ 7.69 (bs, 1H), 7.3-7.2 (d, 2H), 7.05-6.9 (m, 3H), 6.8-6.6 (m, 6H), 3.67 (s, 3H), 3.55 (d, 1H), 3.28 (d, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and methylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.99 (bs, 1H), 7.15 (m, 1H), 5.98 (m, 1H), 6.03 (s, 1H), 1.37 (s, 3H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and cyclopentylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.98 (bs, 1H), 7.15 (m, 1H), 5.94 (m, 1H), 2.31 (m, 1H), 1.7-1.3 (m, 7H), 1.15 (m, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and cyclohexylmethylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.01 (bs, 1H), 5.96 (m, 1H), 6.99 (m, 1H), 1.74 (d, 2H), 1.6-1.3 (m, 5H), 1.1-0.7 (m, 6H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and pyridine-4-ylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.37 (bs, 1H), 8.53 (m, 2H), 7.27 (m, 2H), 7.08 (s, 1H), 7.06-6.9 (m, 2H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and isopropylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.00 (bs, 1H), 7.1-6.9 (m, 2H), 5.99 (s, 1H), 2.08 (m, 1H), 0.95 (d, 3H), 0.66 (d, 3H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and thiophen-2-ylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.23 (bs, 1H), 7.91 (d, 1H), 7.52 (dd, 1H), 7.16 (m, 1H), 7.06 (m, 1H) 7.05 (s, 1H), 6.95 (dd, 1H), 6.71 (dd, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and n-butylylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.00 (bs, 1H), 7.09 (m, 1H), 6.98 (s, 1H), 6.01 (m, 1H), 1.77 (m, 2H), 1.18 (m, 2H), 1.08-0.84 (m, 2H), 0.78 (t, 3H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and cyclohexylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.98 (bs, 1H), 7.06 (m, 1H), 6.97 (m, 1H), 5.94 (s, 1H), 0.75-0.6 (m, 6H), 1.2-0.9 (m, 4H), 1.08-0.84 (m, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and n-propylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.00 (bs, 1H), 7.09 (m, 1H), 6.98 (m, 1H), 6.01 (s, 1H), 1.75 (m, 2H), 1.1-0.9 (m, 2H), 0.78 (t, 3H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and n-pentylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.00 (bs, 1H), 7.09 (m, 1H), 6.98 (m, 1H), 6.01 (s, 1H), 1.76 (m, 2H), 1.25-0.9 (m, 2H), 0.78 (t, 3H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and thiophen-3-ylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.16 (bs, 1H), 7.50 dd, 1H), 7.24 (dd, 1H), 7.15-6.9 (m, 3H), 6.76 (s, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and pyridin-3-ylmagnesium bromide. 1H-NMR (CDCl3) δ 8.61 (bs, 1H), 8.37 (1H), 8.15 (1H, bs), 7.69 (1H), 7.16 (1H), 6.89 (m, 1H), 6.73 (m, 1H), 5.23 (s, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and but-3-en-2-ylmagnesium chloride.
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and sec-butylmagnesium chloride.
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and cycloheptylmagnesium bromide.
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and (1-(benzyloxy)-1H-pyrazol-4-yl)magnesium bromide (J. Org. Chem. (1999) 64, 4196-4198). 1H-NMR (CDCl3) δ 9.01 (bs, 1H), 7.4-7.2 (m, 7H), 7.04 (m, 2H), 6.84 (m, 1H), 5.19 (s, 2H).
General procedure 1. Starting materials: 7-(trifluoromethyl)indoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (CDCl3) δ 7.86 (bs, 1H), 7.52 (dd, 2H), 7.18 (d, 1H), 2.92 (s, 1H), 2.0-1.5 (m, 6H), 1.40-1.0 (m, 4H), 0.86 (m, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and (3,4-difluorophenyl)magnesium bromide. 1H-NMR (DMSO-d6) δ 13.74 (bs, 1H), 8.1 (m, 2H), 7.73 (m, 1H), 7.49 (m, 2H), 6.99 (s, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and (3-fluoro-4-methylphenyl)magnesium bromide. 1H-NMR (DMSO-d6) δ 11.20 (bs, 1H), 7.21 (m, 1H), 7.11 (m, 1H), 7.03-6.78 (m, 6H), 2.19 (s, 3H).
General procedure 1. Starting materials: 6-chloro-7-methylindoline-2,3-dione and cyclohexylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.47 (bs, 1H), 7.04 (m, 2H), 5.79 (s, 1H), 2.21 (s, 3H), 1.9-1.4 (m, 6H), 1.2-0.85 (m, 4H), 0.65 (m, 1H).
General procedure 1. Starting materials: 6,7-dimethylindoline-2,3-dione and cyclohexylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.14 (bs, 1H), 6.93 (d, 1H), 6.6 (d, 1H), 5.56 (s, 1H), 2.19 (s, 3H) 2.08 (s, 3H), 1.9-1.4 (m, 6H), 1.2-0.85 (m, 4H), 0.63 (m, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and (cyclopentylmethyl)magnesium bromide. 1H-NMR (DMSO-d6) δ 11.01 (bs, 1H), 7.10 (m, 1H), 6.98 (m, 1H), 5.98 (s, 1H), 1.93 (m, 2H), 1.65-1.2 (m, 7H), 1.0 (m, 1H), 0.84 (m, 1H).
General procedure 2 using o-cresol instead of phenol. Starting materials: compound 2. 1H-NMR (CDCl3) δ 7.78 (bs, 1H), 7.08 (d, 2H), 6.95-6.7 (m, 6H), 6.59 (d, 2H), 3.71 (s, 3H), 2.10 (s, 3H).
General procedure 2 using m-cresol instead of phenol. Starting materials: compound 2. 1H-NMR (DMSO-d6) δ 11.05 (bs, 1H), 9.47 (bs, 1H), 7.22 (d, 2H), 6.93 (m, 3H), 6.51 (m, 3H), 3.75 (s, 3H), 2.17 (s, 3H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and cyclooctylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.02 (bs, 1H), 7.10 (m, 1H), 6.95 (m, 1H), 2.04 (m, 1H), 1.76-1.20 (m, 13H), 0.86 (m, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and naphthalene-1-ylmagnesium bromide. 1H-NMR (CDCl3) δ 8.23 (bs, 1H), 7.98-7.8 (m, 4H), 7.55-7.4 (m, 3H), 7.28 (s, 1H), 6.96 (m, 1H), 6.76 (m, 1H).
General procedure 1. Starting materials: 6,7-difluoroindoline-2,3-dione and naphthalene-2-ylmagnesium bromide. 1H-NMR (DMSO-d6) δ 11.29 (bs, 1H), 8.0-7.8 (m, 4H), 7.52 (m, 2H), 7.36 (dd, 1H), 7.06-6.95 (m, 3H).
General procedure 1. Starting materials: 7-fluoro-6-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (CDCl3) δ 8.38 (bs, 1H), 7.07 (d, 1H), 6.87 (t, 1H), 5.40 (bs, 1H), 2.28 (m, 4H), 2.15-1.3 (m, 11H), 0.90 (m, 1H).
General procedure 1. Starting materials: 7-fluoro-6-methylindoline-2,3-dione and cyclohexylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.62 (bs, 1H), 6.96 (d, 1H), 6.29 (t, 1H), 5.80 (s, 1H), 2.22 (d, 3H), 1.9-1.4 (m, 6H), 1.25-0.85 (m, 4H), 0.62 (m, 1H).
General procedure 1. Starting materials: 6,7-difluoro-indoline-2,3-dione and tert-butylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.97 (bs, 1H), 7.14 (m, 1H), 7.00 (m, 1H), 5.93 (s, 1H), 1.00 (s, 3H).
General procedure 1. Starting materials: 6,7-difluoro-indoline-2,3-dione and 1,1-dimethylpropylbutylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.97 (bs, 1H), 7.07 (m, 1H), 6.94 (m, 1H), 5.87 (s, 1H), 1.3 (m, 2H), 0.92 (s, 6H), 0.75 (t, 3H).
General procedure 1. Starting materials: 6-fluoro-7-methylindoline-2,3-dione and cyclopentylmagnesium bromide. 1H-NMR (CDCl3) δ 8.72 (bs, 1H), 7.05 (m, 1H), 6.61 (t, 1H), 2.38 (m, 1H), 2.09 (s, 3H), 1.8-1.3 (m, 7H), 1.07 (m, 1H).
General procedure 1. Starting materials: 6-fluoro-7-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (CDCl3) δ 9.32 (bs, 1H), 7.14 (m, 1H), 6.73 (t, 1H), 2.17 (s, 3H), 2.05-0.95 (m, 10H), 0.78 (m, 1H).
General procedure 1. Starting materials: 6-fluoro-7-methylindoline-2,3-dione and n-pentylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.47 (bs, 1H), 7.08 (m, 1H), 6.73 (m, 1H), 5.81 (bs, 1H), 2.12 (s, 3H), 1.17 (m, 2H), 1.4-0.8 (m, 6H), 0.78 (t, 3H).
General procedure 1. Starting materials: 6-fluoro-7-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (CDCl3) δ 8.58 (bs, 1H), 7.18 (m, 1H), 6.38 (m, 1H), 2.85 (m, 1H), 2.20 (s, 3H), 2.3-1.3 (m, 11H), 0.90 (m, 1H).
General procedure 1. Starting materials: 7-(trifluoromethyl)indoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.73 (bs, 1H), 7.51 (m, 2H), 7.10 (t, 1H), 6.04 (s, 1H), 2.2-1.1 (m, 12H), 0.80 (m, 1H).
General procedure 1. Starting materials: 6,7-dimethylindoline-2,3-dione and cycloheptylmagnesium bromide.
General procedure 1. Starting materials: 6-chloro-7-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.49 (bs, 1H), 7.50 (d, 2H), 7.01 (d, 1H), 5.85 (s, 1H), 2.22 (s, 3H), 2.1-1.1 (m, 12H), 0.79 (m, 1H).
General procedure 1. Starting materials: 6-methoxy-7-methylindoline-2,3-dione and cyclopentylmagnesium bromide.
General procedure 1. Starting materials: 6-methoxy-7-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.19 (bs, 1H), 6.99 (d, 1H), 6.51 (d, 1H), 5.52 (s, 1H), 3.76 (s, 3H), 2.01 (s, 3H), 1.95-1.4 (m, 6H), 1.25-0.85 (m, 4H), 0.63 (m, 1H).
General procedure 1. Starting materials: 6-methoxy-7-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.22 (bs, 1H), 7.01 (d, 1H), 6.50 (d, 1H), 5.59 (s, 1H), 3.76 (s, 3H), 2.01 (s, 3H), 2.2-1.2 (m, 13H), 0.75 (m, 1H).
General procedure 1. Starting materials: 7-(trifluoromethyl)indoline-2,3-dione and (4-(benzyloxy)phenyl)magnesium bromide. 1H-NMR (DMSO-d6) δ 10.85 (bs, 1H), 7.55 (d, 1H), 7.5-7.28 (m, 6H), 7.18 (m, 3H), 6.98 (m, 2H), 6.75 (s, 1H), 5.08 (s, 2H).
Imidazole (204 mg, 3 mmol) was dissolved in DCM (dried), cooled to 0° C., and thionylchloride (55 μl, 0.75 mmol) was added with stirring. After 30 minutes compound 46 (200 mg, 0.5 mmol) was added. After a further 2 h the reaction mixture was extracted with H2O, brine, dried over Mg2SO4, filtered and concentrated. The residue was purified by chromatography (1% methanol in DCM) to afford compound 47. 1H-NMR (DMSO-d6) δ 11.59 (bs, 1H), 7.82 (d, 1H), 7.69 (d, 1H), 7.59 (s, 1H), 7.5-7.2 (m, 6H), 7.15-6.95 (m, 6H), 5.10 (s, 2H).
General procedure 1. Starting materials: 6,7-difluoromethylindoline-2,3-dione and (4-(benzyloxy)phenyl)magnesium bromide. 1H-NMR (DMSO-d6) δ 11.13 (bs, 1H), 7.5-7.28 (m, 5H), 7.19 (m, 2H), 7.05-6.90 (m, 4H), 6.72 (s, 1H), 5.08 (s, 2H).
Imidazole (204 mg, 3 mmol) was dissolved in DCM (dried), cooled to 0° C., and thionylchloride (55 μl, 0.75 mmol) was added with stirring. After 30 minutes compound 48 (200 mg, 0.5 mmol) was added. After a further 2 h the reaction mixture was extracted with H2O, brine, dried over Mg2SO4, filtered and concentrated. The residue was purified by chromatography (1% methanol in DCM) to afford compound 49. 1H-NMR (CDCl3) δ 8.94 (bs, 1H), 7.59 (s, 1H), 7.5-7.3 (m, 5H), 7.25-7.1 (m, 3H), 7.09-6.85 (m, 5H), 5.08 (s, 2H).
6,7-Difluoro-3-hydroxy-3-(4-methoxyphenyl)indolin-2-one (WO2005097107) (245 mg, 0.84 mmol) was dissolved in DCM (dried), cooled to 0° C., pyridine (82 μl, 1.01 mmol) and thionylchloride (74 μl, 1.01 mmol) were added with stirring. After 2 h, morpholine (73 μl, 1.01 mmol) and DIEA (398 μl, 2.28 mmol) were added, and the mixture allowed to reach rt and stirred overnight. The reaction mixture was extracted with H2O, brine, dried over Mg2SO4, filtered and concentrated. The residue was purified by chromatography (petroleum ether:EtOAc 20:1 to 5:1) to afford compound 50. 1H-NMR (CDCl3) δ 8.50 (bs, 1H), 7.38 (m, 2H), 6.96 (m, 1H), 6.79 (m, 3H), 3.71 (s, 3H), 3.62 (m, 4H), 2.52 (m, 4H).
General procedure 1. Starting materials: 7-(trifluoromethyl)indoline-2,3-dione and thiazol-2-yllithium. 1H-NMR (DMSO-d6) δ 11.08 (bs, 1H), 7.73 (d, 1H), 7.68 (d, 1H), 7.58 (s, 1H), 7.57 (d, 1H), 7.41 (d, 1H), 7.15 (t, 1H).
General procedure 1. Starting materials: 7-chloro-6-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.53 (bs, 1H), 7.09 (d, 1H), 6.94 (d, 1H), 5.84 (s, 1H), 2.20 (s, 3H), 1.9-0.85 (m, 10H), 0.65 (m, 1H).
General procedure 1. Starting materials: 7-chloro-6-methylindoline-2,3-dione and cyclopentylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.53 (bs, 1H), 7.15 (d, 1H), 6.94 (d, 1H), 5.90 (s, 1H), 2.30 (s, 3H), 1.75-1.25 (m, 8H), 1.10 (m, 1H).
General procedure 1. Starting materials: 7-chloro-6-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.55 (bs, 1H), 7.10 (d, 1H), 6.93 (d, 1H), 5.91 (s, 1H), 2.30 (s, 3H), 2.06 (m, 1H), 1.91 (m, 1H), 1.71 (m, 1H), 1.6-1.15 (m, 9H), 0.76 (m, 1H).
General procedure 1. Starting materials: 7-bromo-6-methylindoline-2,3-dione and cyclopentylmagnesium bromide. 1H-NMR (CDCl3) δ 7.79 (bs, 1H), 7.24 (d, 1H), 6.96 (d, 1H), 3.05 (bs, 1H), 2.46 (m, 1H), 2.41 (s, 3H), 1.9-1.4 (m, 7H), 1.27 (m, 1H).
General procedure 1. Starting materials: 7-bromo-6-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.38 (s, 1H), 7.12 (d, 1H), 6.95 (d, 1H), 5.85 (s, 1H), 2.32 (s, 3H), 1.9-0.9 (m, 10H), 0.66 (m, 1H).
General procedure 1. Starting materials: 7-bromo-6-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.41 (s, 1H), 7.13 (d, 1H), 6.94 (d, 1H), 5.91 (s, 1H), 2.32 (s, 3H), 2.06 (m, 1H), 1.90 (m, 1H), 1.71 (m, 1H), 1.6-1.15 (m, 9H), 0.76 (m, 1H).
General procedure 1. Starting materials: 7-methylindoline-2,3-dione and cyclopentylmagnesium bromide. MS [M+Na]+=254.0, [M−H]−=230.0
General procedure 1. Starting materials: 7-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.22 (bs, 1H), 7.03 (m, 2H), 6.86 (t, 1H), 5.66 (s, 1H), 2.19 (m, 1H), 2.17 (s, 3H), 1.9-1.4 (m, 6H), 1.15 (m, 3H), 0.65 (m, 1H).
General procedure 1. Starting materials: 7-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.24 (bs, 1H), 7.04 (m, 2H), 6.85 (t, 1H), 5.73 (s, 1H), 2.17 (s, 3H), 2.07 (m, 1H), 1.80 (m, 1H), 1.72 (m, 1H), 1.6-1.15 (m, 9H), 0.75 (m, 1H).
General procedure 1. Starting materials: 7-bromoindoline-2,3-dione and cyclopentylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.49 (bs, 1H), 7.40 (d, 1H), 7.29 (d, 1H), 6.92 (t, 1H), 5.96 (s, 1H), 2.31 (m, 1H), 1.75-1.3 (m, 7H), 1.17 (m, 1H).
General procedure 1. Starting materials: 7-bromoindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.49 (bs, 1H), 7.40 (d, 1H), 7.23 (d, 1H), 6.92 (t, 1H), 5.91 (s, 1H), 1.9-0.9 (m, 10H), 0.66 (m, 1H).
General procedure 1. Starting materials: 7-bromoindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.52 (bs, 1H), 7.40 (d, 1H), 7.24 (d, 1H), 6.91 (t, 1H), 5.98 (s, 1H), 2.04 (m, 1H), 1.92 (m, 1H), 1.72 (m, 1H), 1.65-1.15 (m, 9H) 0.79 (m, 1H).
General procedure 1. Starting materials: 7-(trifluoromethyl)indoline-2,3-dione and cyclooctylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.73 (bs, 1H), 7.51 (m, 2H), 7.13 (t, 1H), 6.04 (s, 1H), 2.15-1.2 (m, 14H), 0.86 (m, 1H).
General procedure 1. Starting materials: 7-chloro-6-methylindoline-2,3-dione and cyclooctylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.50 (bs, 1H), 7.06 (d, 1H), 6.84 (d, 1H), 5.84 (s, 1H), 2.23 (s, 3H), 2.15-1.1 (m, 14H), 0.78 (m, 1H).
General procedure 1. Starting materials: 5,7-dimethylindoline-2,3-dione and cyclopentylmagnesium bromide. MS [2M+Na]+=513.3, [M−H]−=244.1
General procedure 1. Starting materials: 5,7-dimethylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.11 (bs, 1H), 6.86 (s, 1H), 6.82 (s, 1H), 5.60 (s, 1H), 2.22 (s, 3H), 2.13 (s, 3H), 1.9-0.85 (m, 10H), 0.66 (m, 1H).
General procedure 1. Starting materials: 5,7-dimethylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.14 (bs, 1H), 6.87 (s, 1H), 6.82 (s, 1H), 5.67 (s, 1H), 2.21 (s, 3H), 2.13 (s, 3H), 2.06 (m, 1H), 1.88 (m, 1H), 1.72 (m, 1H), 1.6-1.2 (m, 9H), 0.78 (m, 1H).
General procedure 1. Starting materials: 5-methoxy-7-methylindoline-2,3-dione and cyclopentylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.07 (bs, 1H), 6.70 (d, 1H), 6.62 (d, 1H), 5.71 (s, 1H), 3.69 (s, 3H), 2.29 (m, 1H), 2.16 (s, 3H), 1.65-1.3 (m, 7H), 1.21 (m, 1H).
General procedure 1. Starting materials: 5-methoxy-7-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.07 (bs, 1H), 6.63 (d, 1H), 6.61 (d, 1H), 5.66 (s, 1H), 3.69 (s, 3H), 2.15 (s, 3H), 1.75 (m, 3H), 1.55 (m, 3H), 1.05 (m, 4H), 0.70 (m, 1H).
General procedure 1. Starting materials: 5-methoxy-7-methylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.09 (bs, 1H), 6.64 (d, 1H), 6.61 (d, 1H), 5.73 (s, 1H), 3.68 (s, 3H), 2.16 (s, 3H), 2.02 (m, 1H), 1.89 (m, 1H), 1.71 (m, 1H), 1.6-1.15 (m, 9H), 0.80 (m, 1H).
General procedure 1. Starting materials: 5-chloro-7-methylindoline-2,3-dione and cyclopentylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.40 (bs, 1H), 7.11 (m, 2H), 5.90 (s, 1H), 2.31 (m, 1H), 2.18 (s, 3H), 1.9-1.3 (m, 7H), 1.21 (m, 1H).
General procedure 1. Starting materials: 5-chloro-7-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.40 (bs, 1H), 7.11 (d, 1H), 7.04 (d, 1H), 5.85 (s, 1H), 2.17 (s, 3H), 1.9-1.4 (m, 7H), 1.2-0.9 (m, 3H), 0.70 (m, 1H).
General procedure 1. Starting materials: 5-methoxy-7-methylindoline-2,3-dione and cycloheptylmagnesium bromide. MS [M+Na]+=316.1, [M−H]−=292.2
General procedure 1. Starting materials: 5-fluoro-7-methylindoline-2,3-dione and cyclopentylmagnesium bromide. MS [M+Na]+=272.1, [M−H]−=248.1
General procedure 1. Starting materials: 5-fluoro-7-methylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.28 (bs, 1H), 6.87 (m, 2H), 5.85 (s, 1H), 2.18 (s, 3H), 1.85-1.45 (m, 6H), 1.2-0.9 (m, 4H), 0.69 (m, 1H).
General procedure 1. Starting materials: 5-fluoro-7-methylindoline-2,3-dione and cycloheptylmagnesium bromide. MS [M+Na]+=290.1, [M−H]−=276.1
General procedure 1. Starting materials: 6-fluoro-5,7-dimethylindoline-2,3-dione and cyclopentylmagnesium bromide. MS [M+H]+=264.1, [M−H]−=262.1
General procedure 1. Starting materials: 6-fluoro-5,7-dimethylindoline-2,3-dione and cyclohexylmagnesium chloride. 1H-NMR (DMSO-d6) δ 10.33 (bs, 1H), 6.95 (d, 1H), 5.63 (s, 1H), 2.17 (d, 3H), 2.09 (d, 3H), 1.9-1.4 (m, 6H), 1.2-0.9 (m, 4H), 0.66 (m, 1H).
General procedure 1. Starting materials: 6-fluoro-5,7-dimethylindoline-2,3-dione and cycloheptylmagnesium bromide. 1H-NMR (DMSO-d6) δ 10.35 (bs, 1H), 6.95 (d, 1H), 5.73 (s, 1H), 2.15 (d, 3H), 2.09 (d, 3H), 2.10 (m, 1H), 1.88 (m, 1H), 1.74 (m, 1H), 1.65-1.0 (m, 9H), 0.78 (m, 1H).
General procedure 1. Starting materials: 7-methyl-6-(trifluoromethyl)indoline-2,3-dione and cyclopentylmagnesium bromide. MS [M+Na]+=322.2, [M−H]=298.0.
General procedure 1. Starting materials: 7-methyl-6-(trifluoromethyl)indoline-2,3-dione and cyclohexylmagnesium chloride. MS [M+Na]+=336.2, [M−H]−=312.0.
General procedure 1. Starting materials: 7-methyl-6-(trifluoromethyl)indoline-2,3-dione and cycloheptylmagnesium bromide. MS [M+Na]+=350.2, [M−H]−=326.2.
General procedure 1. Starting materials: 5-methoxy-6,7-dimethylindoline-2,3-dione and cyclopentylmagnesium bromide. MS [2M+H]+=572.9, [M−H]−=274.0.
General procedure 1. Starting materials: 5-methoxy-6,7-dimethylindoline-2,3-dione and cyclohexylmagnesium chloride. MS [2M+Na]+=600.9, [M−H]−=288.1.
General procedure 1. Starting materials: 5-methoxy-6,7-dimethylindoline-2,3-dione and cycloheptylmagnesium bromide. MS [M+H]+=304.2, [M−H]−=302.2.
General procedure 2. Starting materials: compound 1. 1H-NMR (DMSO-d6) δ 11.55 (bs, 1H), 9.57 (s, 1H), 7.12-6.9 (m, 4H), 6.75 (d, 2H), 3.62 (s, 1H).
General procedure 5. Starting materials: compound 4. 1H-NMR (DMSO-d6) δ 11.05 (bs, 1H), 9.51 (s, 1H), 7.3-7.2 (m, 3H), 7.18-7.11 (m, 3H), 7.08 (m, 1H), 6.8 (d, 2H), 3.56 (dd, 2H).
General procedure 2. Starting materials: compound 5. 1H-NMR (DMSO-d6) δ 11.20 (bs, 1H), 9.40 (bs, 1H), 7.05-6.95 (m, 4H), 6.69 (d, 2H), 1.62 (s, 3H).
General procedure 2. Starting materials: compound 6. 1H-NMR (DMSO-d6) δ 11.21 (bs, 1H), 9.38 (bs, 1H), 7.16-7.08 (m, 4H), 7.01 (m, 1H), 6.69 (d, 2H), 2.97 (m, 1H), 1.6-1.2 (m, 7H), 0.94 (m, 1H).
General procedure 2. Starting materials: compound 7. 1H-NMR (DMSO-d6) δ 11.19 (bs, 1H), 9.38 (bs, 1H), 7.12-6.9 (m, 4H), 6.67 (d, 2H), 2.12 (m, 2H), 1.6-1.4 (m, 7H), 1.25 (m, 1H), 1.1-0.7 (m, 6H).
General procedure 2. Starting material: compound 8. 1H-NMR (DMSO-d6) δ 10.82 (bs, 1H), 9.01 (bs, 3H), 7.70 (bs, 2H), 7.68-7.42 (m, 4H), 7.29 (m, 2H).
General procedure 2. Starting material: compound 9. 1H-NMR (CDCl3) δ 7.87 (bs, 1H), 7.16 (d, 2H), 6.97 (m, 1H), 6.84 (m, 1H), 6.66 (2, 2H), 3.1 (bs, 1H), 2.77 (m, 1H) 0.89 (d, 3H), 0.68 (d, 3H).
General procedure 2. Starting material: compound 10. 1H-NMR (DMSO-d6) δ 11.54 (bs, 1H), 9.52 (bs, 1H), 7.51 (d, 1H), 7.2-6.9 (m, 6H), 6.70 (d, 2H).
General procedure 2. Starting material: compound 11. 1H-NMR (DMSO-d6) δ 11.21 (bs, 1H), 9.39 (bs, 1H), 7.1-6.9 (m, 4H), 6.68 (d, 2H), 2.13 (m, 2H), 1.21 (m, 2H), 0.98 (m, 1H), 0.78 (m, 4H).
General procedure 2. Starting material: compound 12. 1H-NMR (CDCl3) δ 8.46 (bs, 1H), 7.21 (d, 2H), 7.05 (m, 1H), 6.92 (m, 1H), 6.75 (d, 2H), 6.13 (bs, 1H), 2.48 (m, 1H) 1.8-1.0 (m, 9H), 0.69 (m, 1H).
General procedure 2. Starting material: compound 13. 1H-NMR (DMSO-d6) δ 11.20 (bs, 1H), 9.38 (bs, 1H), 7.12-7.0 (m, 4H), 6.68 (d, 2H), 2.13 (m, 2H), 1.21 (m, 2H), 1.02 (m, 1H), 0.83 (m, 4H).
General procedure 2. Starting material: compound 14. 1H-NMR (DMSO-d6) δ 11.21 (bs, 1H), 9.39 (bs, 1H), 7.1-6.9 (m, 4H), 6.68 (d, 2H), 2.10 (m, 2H), 1.26-0.94 (m, 5H), 0.78 (m, 4H).
General procedure 2. Starting material: compound 15. 1H-NMR (DMSO-d6) δ 11.20 (bs, 1H), 9.43 (bs, 1H), 7.52 (d, 1H), 7.15 (m, 2H), 7.05 (m, 2H), 6.88 (d, 2H), 6.67 (d, 2H).
General procedure 2. Starting material: compound 16. MS [M+K]+=380.20.
Starting material: example 6. Compound 1006 (26 mg, 0.077 mmol) was dissolved in DCM, and peracetic acid in acetic acid (39%, 0.125 mL) was added and the mixture stirred overnight, concentrated and purified by flash chromatography (chloroform:methanol: 25% ammonia 95:5:0.5) to yield compound 1015 (19 mg). 1H-NMR (DMSO-d6) δ 11.67 (bs, 1H), 9.60 (bs, 1H), 8.18 (d, 2H), 7.25-7.00 (m, 4H), 6.97 (d, 2H), 6.74 (d, 2H). MS [M+H]+=355.20.
General procedure 2. Starting material: compound 17. 1H-NMR (DMSO-d6) δ 11.23+11.16 (s, 1H), 9.40 (s, 1H), 7.18-7.0 (m, 4H), 6.71 (d, 2H), 5.73+5.30 (m, 1H), 4.95 (m, 2H), 3.33 (m, 1H), 0.94+0.76 (d, 3H).
General procedure 2. Starting material: compound 18. 1H-NMR (CDCl3) δ 7.62, 7.14 (m, 3H), 6.95 (m, 1H), 6.83 (m, 1H), 6.69 (d, 2H), 5.0 (s, 1H), 2.45 (m, 1H), 1.5-0.98 (m, 2H), 0.86+0.76 (d, 3H), 0.80 (m, 3H).
General procedure 2. Starting material: compound 19. 1H-NMR (CDCl3) δ 7.51 (bs, 1H), 7.14 (d, 2H), 6.96 (m, 1H), 6.82 (m, 2H), 6.68 (d, 2H), 4.89 (s, 1H) 2.57 (m, 1H), 1.7-1.1 (m, 11H), 0.81 (m, 1H).
General procedure 2. Starting material: compound 20. 1H-NMR (CDCl3) δ 7.96 (bs, 1H), 7.35-7.15 (m, 5H), 7.09 (s, 1H), 6.87 (s, 1H), 6.85-6.65 (m, 4H), 6.60 (d, 2H), 5.23 (s, 2H), 5.19 (s, 1H). Example 20: 6,7-difluoro-3-(1-hydroxy-1H-pyrazol-4-yl)-3-(4-hydroxyphenyl)indolin-2-one (compound 1020)
Starting material: Example 19. Compound 1019 (40 mg, 0.09 mmol) was dissolved in methanol (4 mL) and the solution bubbled through with N2 for 2 minutes. 10% Pd/C (3.1 mg) was added. The flask was fitted with a septum and a N2-filled balloon, carefully evacuated and filled with N2. The N2-filled balloon was substituted with a H2-filled balloon, the flask was then carefully evacuated and filled with H2 twice and the reaction mixture vigorously stirred at 0° C. for 30 minutes. The flask was the carefully evacuated and filled with N2 twice, the reaction mixture filtered through Celite, concentrated and purified by flash chromatography (chloroform:methanol: 25% ammonia 80:20:1) to yield Example 20 (26 mg). 1H-NMR (DMSO-d6) δ 7.33 (s, 1H), 7.14 (m, 1H), 7.01 (m, 1H), 6.95 (s, 1H), 6.87 (d, 2H), 6.66 (d, 2H), 4-3 (bs, >3H). MS [M+H]+=344.15.
General procedure 2. Starting material: compound 21. 1H-NMR (CDCl3) δ 8.02 (bs, 1H), 7.52 (m, 2H), 7.23 (m, 3H), 6.87 (s, 1H), 6.76 (d, 2H), 5.91 (bs, 1H), 2.52 (m, 1H), 2.15-0.95 (m, 9H), 0.70 (m, 1H).
General procedure 2. Starting material: compound 22. 1H-NMR (CDCl3) δ 7.96 (bs, 1H), 7.1-6.76 (m, 7H), 6.69 (d, 2H), 5.05 (s, 1H).
General procedure 2. Starting material: compound 23. 1H-NMR (DMSO-d6) δ 11.48 (bs, 1H), 9.49 (s, 1H), 7.24 (m, 1H), 7.14-6.8 (m, 6H), 6.72 (d, 2H), 2.20 (s, 3H).
General procedure 2. Starting material: compound 24. 1H-NMR (DMSO-d6) δ 10.67 (bs, 1H), 9.35 (s, 1H), 7.12 (m, 4H), 6.69 (d, 2H), 2.32 (m, 1H), 2.25 (s, 3H), 1.59 (m, 3H), 1.40 (m, 2H), 1.13 (m, 3H), 0.96 (m, 1H), 0.73 (m, 1H).
General procedure 2. Starting material: compound 25. 1H-NMR (DMSO-d6) δ 10.35 (bs, 1H), 9.29 (s, 1H), 7.13 (m, 2H), 7.01 (d, 1H), 6.85 (d, 1H), 6.68 (d, 2H), 2.25 (m, 1H), 2.23 (s, 3H), 2.12 (s, 3H), 1.75-0.8 (m, 9H), 0.67 (m, 1H).
General procedure 2. Starting material: compound 26. 1H-NMR (DMSO-d6) δ 11.21 (bs, 1H), 9.39 (s, 1H), 7.05 (m, 4H), 6.68 (d, 2H), 2.29 (m, 2H), 1.6-1.0 (m, 8H), 0.86 (m, 1H).
General procedure 6 (heptane:ethanol 70:30, 7 mL/min). Starting material: compound 1010. tR (Chiralcel OD 250×4.6 mm ID 5 micron, heptane:ethanol 70:30, 0.6 mL/min): 8.9 min.
General procedure 6. Starting material: Compound 1010. tR (Chiralcel OD 250×4.6 mm ID 5 micron, heptane:ethanol 70:30, 0.6 mL/min): 14.5 min.
General procedure 5. Starting material: compound 27. 1H-NMR (DMSO-d6) δ 11.34 (bs, 1H), 9.29 (bd, 2H), 7.05-6.9 (m, 4H), 6.83 (d, 1H), 6.8-6.6 (m, 4H), 2.03 (s, 3H).
General procedure 5. Starting material: compound 28.
1H-NMR (DMSO-d6) δ 10.98 (bs, 1H), 9.46 (bs, 1H), 9.40 (bs), 7.11 (d, 2H), 6.92 (m, 1H), 6.73 (m, 3H), 6.52 (m, 3H), 2.17 (s, 3H).
General procedure 2. Starting material: compound 29. 1H-NMR (DMSO-d6) δ 11.16 (bs, 1H), 9.40 (bs, 1H), 7.20-6.90 (m, 4H), 7.05 (d, 2H), 2.65 (m, 1H), 1.7-1.2 (m, 13H), 0.89 (m, 1H).
General procedure 2. Starting material: compound 30. 1H-NMR (DMSO-d6) δ 11.55 (bs, 1H), 9.53 (bs, 1H), 7.92 (t, 2H), 7.6-7.2 (m, 4H), 7.2-6.8 (m, 5H), 6.72 (d, 2H).
General procedure 2. Starting material: compound 31. 1H-NMR (DMSO-d6) δ 11.55 (bs, 1H), 9.53 (bs, 1H), 7.92-7.80 (m, 3H), 7.62 (s, 1H), 7.50 (m, 2H), 7.34 (dd, 1H), 7.15 (m, 1H), 7.06 (m, 1H), 7.00 (d, 2H), 6.74 (d, 2H).
General procedure 2. Starting material: compound 32. 1H-NMR (CDCl3) δ 7.64 (bs, 1H), 7.13 (d, 2H), 6.92 (d, 1H), 6.82 (t, 1H), 6.64 (d, 2H), 5.51 (bs, 1H), 2.56 (m, 1H), 2.26 (d, 3H), 1.7-1.3 (m, 10H), 0.80 (m, 2H).
General procedure 2. Starting material: compound 33. 1H-NMR (DMSO-d6) δ 10.82 (bs, 1H), 9.33 (bs, 1H), 7.12 (m, 2H), 7.04 (d, 1H), 6.92 (t, 1H), 6.70 (m, 2H), 2.30 (m, 4H), 1.7-0.8 (m, 9H), 0.68 (m, 1H).
General procedure 2. Starting material: compound 34. 1H-NMR (DMSO-d6) δ 11.32 (bs, 1H), 9.41 (bs, 1H), 7.63 (m, 3H), 7.03 (m, 1H), 6.71 (m, 2H), 0.95 (s, 3H).
General procedure 2. Starting material: compound 35. 1H-NMR (DMSO-d6) δ 11.32 (bs, 1H), 9.41 (bs, 1H), 7.63 (m, 3H), 7.01 (m, 1H), 6.71 (m, 2H), 1.3 (m, 2H), 0.94 (d, 6H), 0.69 (t, 3H).
General procedure 2. Starting material: compound 36. 1H-NMR (CDCl3) δ 8.5 (bs, 1H), 7.16 (m, 2H), 7.00 (m, 1H), 6.6 (m, 3H), 5.5 (bs, 1H), 2.96 (m, 1H), 2.12 (s, 3H), 1.7-0.65 (m, 9H).
General procedure 2. Starting material: compound 37. 1H-NMR (DMSO-d6) δ 10.65 (bs, 1H), 9.33 (bs, 1H), 7.13 (m, 3H), 6.80 (m, 1H), 6.69 (m, 2H), 2.31 (m, 1H), 2.15 (s, 3H), 1.7-0.8 (m, 9H), 0.70 (m, 1H).
General procedure 2. Starting material: compound 38. 1H-NMR (CDCl3) δ 10.68 (bs, 1H), 9.34 (bs, 1H), 7.06 (m, 2H), 7.02 (m, 1H), 6.77 (m, 1H), 6.67 (m, 2H), 2.17 (s, 3H), 2.08 (m, 2H), 1.19 (m, 4H), 1.00 (m, 1H), 0.9-0.7 (m, 4H).
General procedure 2. Starting material: compound 39. 1H-NMR (DMSO-d6) δ 10.63 (bs, 1H), 9.35 (bs, 1H), 7.1 (m, 3H), 6.80 (m, 1H), 6.70 (m, 2H), 2.47 (m, 1H), 2.14 (s, 3H), 1.7-1.1 (m, 11H), 0.85 (m, 1H).
General procedure 2. Starting material: compound 40. 1H-NMR (DMSO-d6) δ 10.90 (bs, 1H), 9.42 (bs, 1H), 7.56 (m, 2H), 7.21 (m, 1H), 7.09 (m, 2H), 6.71 (m, 2H), 2.55 (m, 1H), 1.7-1.2 (m, 11H), 0.86 (m, 1H).
General procedure 2. Starting material: compound 41. 1H-NMR (DMSO-d6) δ 10.31 (bs, 1H), 9.29 (bs, 1H), 7.10 (m, 2H), 6.99 (d, 1H), 6.84 (d, 1H), 6.67 (m, 2H), 2.46 (m, 1H), 2.23 (s, 3H), 2.12 (s, 3H), 1.65-1.2 (m, 11H), 0.85 (m, 1H).
General procedure 2. Starting material: compound 42. 1H-NMR (DMSO-d6) δ 10.63 (bs, 1H), 9.36 (bs, 1H), 7.09 (m, 4H), 6.69 (m, 2H), 2.26 (s, 3H), 1.7-1.15 (m, 12H), 0.85 (m, 1H).
General procedure 2. Starting material: compound 43. 1H-NMR (DMSO-d6) δ 10.40 (bs, 1H), 9.27 (bs, 1H), 7.13 (m, 2H), 7.05 (d, 1H), 6.67 (m, 2H), 6.58 (d, 1H), 3.78 (s, 3H), 2.93 (m, 1H), 2.07 (s, 3H), 1.6-1.2 (m, 7H), 0.86 (m, 1H).
General procedure 2. Starting material: compound 44. 1H-NMR (DMSO-d6) δ 10.39 (bs, 1H), 9.28 (bs, 1H), 7.14 (m, 2H), 7.07 (d, 1H), 6.68 (m, 2H), 6.61 (d, 1H), 3.79 (s, 3H), 2.28 (m, 1H), 2.05 (s, 3H), 1.55 (m, 3H), 1.4 (m, 2H), 1.16 (m, 3H), 0.96 (m, 1H), 0.69 (m, 1H).
Compound 47 (184 mg, 0.36 mmol) was dissolved in methanol (10 mL) and the solution bubbled through with N2 for 2 minutes. 10% Pd/C (60 mg) was added. The flask was fitted with a septum and a N2-filled balloon, carefully evacuated and filled with N2. The N2-filled balloon was substituted with a H2-filled balloon, the flask was then carefully evacuated and filled with H2 twice and the reaction mixture vigorously stirred at rt for 4 h. The flask was the carefully evacuated and filled with N2 twice, the reaction mixture filtered through Celite, concentrated and purified by flash chromatography (1%-5% methanol in DCM) to yield compound 1047. 1H-NMR (DMSO-d6) δ 11.53 (bs, 1H), 9.79 (bs, 1H), 7.78 (d, 1H), 7.68 (d, 1H), 7.56 (s, 1H), 7.28 (t, 1H), 7.1-6.9 (m, 4H), 6.80 (d, 2H).
Compound 49 (127 mg, 0.30 mmol) was dissolved in methanol (10 mL) and the solution bubbled through with N2 for 2 minutes. 10% Pd/C (46 mg) was added. The flask was fitted with a septum and a N2-filled balloon, carefully evacuated and filled with N2. The N2-filled balloon was substituted with a H2-filled balloon, the flask was then carefully evacuated and filled with H2 twice and the reaction mixture vigorously stirred at rt for 4 h. The flask was the carefully evacuated and filled with N2 twice, the reaction mixture filtered through Celite, concentrated and purified by flash chromatography (1%-5% methanol in DCM) to yield compound 1048. 1H-NMR (DMSO-d6) δ 11.84 (bs, 1H), 9.78 (bs, 1H), 7.54 (s, 1H), 7.34 (m, 1H), 7.2-7.07 (m, 2H), 7.05-6.95 (m, 3H), 6.79 (m, 2H).
Compound 50 (82 mg, 0.23 mmol) was dissolved in DCM under nitrogen, cooled to −78° C. and BBr3-solution (1.0 M, 341 μl, 0.35 mmol) was added dropwise with stirring. The reaction mixture was gradually allowed to reach room temperature and stirred on. The resulting precipitate was filtered off, washed with DCM and purified by chromatography (CHCl3:MeOH:NH3 (25%) 98:2:0.2) to afford compound 1049. 1H-NMR (MeOD) δ 7.23 (m, 2H), 7.01 (m, 1H), 6.83 (m, 1H), 6.66 (m, 2H), 3.57 (m, 4H), 2.45 (m, 2H).
General procedure 2. Starting material: compound 51. 1H-NMR (DMSO-d6) δ 11.36 (bs, 1H), 9.64 (bs, 1H), 7.80 (d, 1H), 7.75 (d, 1H), 7.70 (d, 1H), 7.59 (d, 1H), 7.59 (d, 1H), 7.23 (t, 1H), 7.07 (m, 2H), 6.75 (m, 2H).
General procedure 2. Starting material: compound 52. 1H-NMR (DMSO-d6) δ 10.72 (s, 1H), 9.35 (bs, 1H), 7.18 (d, 1H), 7.12 (m, 2H), 7.02 (d, 1H), 6.70 (m, 2H), 2.34 (s, 3H), 2.28 (m, 1H), 1.60 (m, 3H), 1.41 (m, 2H), 1.3-0.8 (m, 4H), 0.70 (m, 1H).
General procedure 2. Starting material: compound 53. 1H-NMR (DMSO-d6) δ 10.74 (s, 1H), 9.35 (s, 1H), 7.16 (d, 1H), 7.12 (m, 2H), 6.99 (d, 1H), 6.69 (m, 2H), 2.95 (m, 1H), 2.33 (s, 3H), 1.65-1.2 (m, 7H), 0.91 (m, 1H).
General procedure 2. Starting material: compound 54. 1H-NMR (DMSO-d6) δ 10.70 (s, 1H), 9.36 (s, 1H), 7.16 (d, 1H), 7.09 (m, 2H), 7.02 (d, 1H), 6.69 (m, 2H), 2.47 (m, 1H), 2.34 (s, 3H), 1.65-1.2 (m, 11H), 0.85 (m, 1H).
Compound 1046 was suspended in 37% aqueous HBr and heated to 120° C. in a microwave oven for 3 times 10 minutes, concentrated twice with toluene and purified by chromatography (1%-5% MeOH in DCM) to afford compound 1054. MS [M+H]+=338.1 [M−H]+=336.1
General procedure 2. Starting material: compound 55. 1H-NMR (DMSO-d6) δ 10.60 (s, 1H), 9.35 (s, 1H), 7.19 (d, 1H), 7.12 (m, 2H), 6.99 (m, 1H), 6.69 (m, 2H), 2.95 (m, 1H), 2.35 (s, 3H), 1.6-1.2 (m, 7H), 0.92 (m, 1H).
General procedure 2. Starting material: compound 56. 1H-NMR (DMSO-d6) δ 10.57 (s, 1H), 9.36 (s, 1H), 7.21 (d, 1H), 7.12 (m, 2H), 7.03 (m, 1H), 6.70 (m, 2H), 2.35 (s, 3H), 2.30 (m, 1H), 1.59 (m, 3H), 1.41 (m, 2H), 1.16 (m, 3H), 0.96 (m, 1H), 0.70 (m, 1H).
General procedure 2. Starting material: compound 57. 1H-NMR (DMSO-d6) δ 10.55 (s, 1H), 9.36 (s, 1H), 7.19 (d, 1H), 7.09 (m, 2H), 7.02 (m, 1H), 6.70 (m, 2H), 2.49 (m, 1H), 2.36 (s, 3H), 1.7-1.2 (m, 11H), 0.85 (m, 1H).
General procedure 2. Starting material: compound 58. 1H-NMR (MeOD) δ 7.19 (m, 2H), 7.12 (m, 2H), 6.99 (t, 1H), 6.71 (m, 2H), 3.07 (m, 1H), 2.31 (s, 3H), 1.8-1.2 (m, 7H), 0.94 (m, 1H). MS [M+Na]+=330.1, [M−H]−=306.1
General procedure 2. Starting material: compound 59. 1H-NMR (DMSO-d6) δ 10.42 (bs, 1H), 9.32 (bs, 1H), 7.14 (m, 3H), 7.05 (d, 1H), 6.94 (t, 1H), 6.69 (m, 2H), 2.30 (m, 1H), 2.21 (s, 3H), 1.7-0.8 (m, 9H), 0.70 (m, 1H).
General procedure 2. Starting material: compound 60. 1H-NMR (DMSO-d6) δ 10.39 (bs, 1H), 9.31 (bs, 1H), 7.11 (m, 3H), 7.05 (d, 1H), 6.93 (t, 1H), 6.68 (m, 2H), 2.46 (m, 1H), 2.21 (s, 3H), 1.7-1.2 (m, 9H), 0.85 (m, 1H).
General procedure 2. Starting material: compound 61. 1H-NMR (DMSO-d6) δ 10.73 (s, 1H), 9.37 (s, 1H), 7.44 (d, 1H), 7.30 (d, 1H), 7.12 (m, 2H), 6.97 (t, 1H), 6.70 (m, 2H), 2.97 (m, 1H), 1.6-1.2 (m, 7H), 0.98 (m, 1H).
General procedure 2. Starting material: compound 62. 1H-NMR (DMSO-d6) δ 10.70 (s, 1H), 9.38 (s, 1H), 7.50 (d, 1H), 7.33 (d, 1H), 7.12 (m, 2H), 7.00 (t, 1H), 6.71 (m, 2H), 2.33 (m, 1H), 1.62 (m, 3H), 1.41 (m, 2H), 1.16 (m, 3H) 0.97 (m, 1H), 0.74 (m, 1H).
General procedure 2. Starting material: compound 63. 1H-NMR (DMSO-d6) δ 10.67 (bs, 1H), 9.38 (s, 1H), 7.45 (d, 1H), 7.30 (d, 1H), 7.09 (m, 2H), 6.99 (t, 1H), 6.70 (m, 2H), 2.53 (m, 1H), 1.7-1.2 (m, 11H), 0.86 (m, 1H).
General procedure 2. Starting materials: compound 64. 1H-NMR (DMSO-d6) δ 10.89 (bs, 1H), 9.41 (s, 1H), 7.63 (d, 1H), 7.54 (d, 1H), 7.21 (t, 1H), 7.11 (m, 2H), 6.72 (m, 2H), 2.68 (m, 1H), 1.7-1.1 (m, 13H), 0.93 (m, 1H).
General procedure 2. Starting materials: compound 65. 1H-NMR (DMSO-d6) δ 10.70 (bs, 1H), 9.37 (s, 1H), 7.18 (d, 1H), 7.11 (m, 2H), 7.01 (d, 1H), 6.70 (m, 2H), 2.62 (m, 1H), 2.33 (s, 3H), 1.65-1.05 (m, 13H), 0.87 (m, 1H).
General procedure 2. Starting material: compound 66. 1H-NMR (DMSO-d6) δ 10.35 (bs, 1H), 9.29 (s, 1H), 7.13 (m, 2H), 6.90 (s, 1H), 6.84 (s, 1H), 6.68 (m, 2H), 2.92 (m, 1H), 2.24 (s, 3H), 2.19 (s, 3H), 1.6-1.2 (m, 7H), 0.98 (m, 1H).
General procedure 2. Starting material: compound 67. 1H-NMR (DMSO-d6) δ 10.32 (bs, 1H), 9.29 (s, 1H), 7.14 (m, 2H), 6.93 (s, 1H), 6.85 (s, 1H), 6.68 (m, 2H), 2.30 (m, 1H), 2.27 (s, 3H), 2.17 (s, 3H), 1.60 (m, 3H), 1.41 (m, 2H), 1.15 (m, 3H), 0.97 (m, 1H), 0.76 (m, 1H).
General procedure 2. Starting material: compound 68. 1H-NMR (DMSO-d6) δ 10.29 (bs, 1H), 9.32 (bs, 1H), 7.11 (m, 2H), 6.91 (s, 1H), 6.85 (s, 1H), 6.68 (m, 2H), 2.47 (m, 1H), 2.26 (s, 3H), 2.17 (s, 3H), 1.7-1.1 (m, 11H), 0.86 (m, 1H).
General procedure 2. Starting material: compound 69. 1H-NMR (DMSO-d6) δ 10.30 (bs, 1H), 9.30 (bs, 1H), 7.14 (m, 2H), 6.67 (m, 4H), 3.69 (s, 3H), 2.95 (m, 1H), 2.21 (s, 3H), 1.6-1.2 (m, 7H), 1.00 (m, 1H).
General procedure 2. Starting material: compound 70. 1H-NMR (DMSO-d6) δ 10.27 (bs, 1H), 9.30 (bs, 1H), 7.14 (m, 2H), 6.68 (m, 4H), 3.71 (s, 3H), 2.30 (m, 1H), 2.20 (s, 3H), 1.59 (m, 3H), 1.39 (m, 2H), 1.15 (m, 3H), 0.98 (m, 1H), 0.77 (m, 1H).
General procedure 2. Starting material: compound 71. 1H-NMR (DMSO-d6) δ 10.29 (bs, 1H), 9.31 (bs, 1H), 7.11 (m, 2H), 6.68 (m, 4H), 3.71 (s, 3H), 2.45 (m, 1H), 2.20 (s, 3H), 1.7-1.15 (m, 11H), 0.87 (m, 1H).
General procedure 2. Starting material: compound 72. 1H-NMR (DMSO-d6) δ 10.65 (bs, 1H), 9.35 (bs, 1H), 7.13 (m, 4H), 6.70 (m, 2H), 2.97 (m, 1H), 2.23 (s, 3H), 1.65-1.35 (m, 6H), 1.24 (m, 1H), 0.90 (m, 1H).
General procedure 2. Starting material: compound 73. 1H-NMR (DMSO-d6) δ 10.62 (bs, 1H), 9.36 (bs, 1H), 7.13 (m, 4H), 6.71 (m, 2H), 2.35 (m, 1H), 2.21 (s, 3H), 1.61 (m, 3H), 1.39 (m, 2H), 1.3-0.9 (m, 4H), 0.81 (m, 1H).
General procedure 2. Starting material: compound 74. 1H-NMR (DMSO-d6) δ 10.59 (bs, 1H), 9.38 (s, 1H), 7.11 (m, 4H), 6.71 (m, 2H), 2.49 (m, 1H), 2.21 (s, 3H), 1.7-1.15 (m, 11H), 0.89 (m, 1H).
General procedure 2. Starting material: compound 75. 1H-NMR (DMSO-d6) δ 10.52 (bs, 1H), 9.33 (bs, 1H), 7.12 (m, 2H), 6.95 (dd, 1H), 6.89 (dd, 1H), 6.70 (m, 2H), 2.95 (m, 1H), 2.22 (s, 3H), 1.6-0.95 (m, 7H), 0.87 (m, 1H).
General procedure 2. Starting material: compound 76. 1H-NMR (DMSO-d6) δ 10.49 (bs, 1H), 9.34 (bs, 1H), 7.14 (m, 2H), 7.01 (dd, 1H), 6.93 (dd, 1H), 6.70 (m, 2H), 2.32 (m, 1H), 2.22 (s, 3H), 1.60 (m, 3H), 1.40 (m, 2H), 1.3-0.9 (m, 4H), 0.78 (m, 1H).
General procedure 2. Starting material: compound 77. 1H-NMR (DMSO-d6) δ 10.46 (bs, 1H), 9.35 (s, 1H), 7.11 (m, 2H), 6.95 (m, 2H), 6.70 (m, 2H), 2.49 (m, 1H), 2.22 (s, 3H), 1.7-1.1 (m, 11H), 0.87 (m, 1H).
General procedure 2. Starting material: compound 78. 1H-NMR (DMSO-d6) δ 10.55 (bs, 1H), 9.32 (bs, 1H), 7.12 (m, 2H), 7.00 (d, 1H), 6.68 (m, 2H), 2.92 (m, 1H), 2.18 (d, 3H), 2.15 (d, 3H), 1.6-1.2 (m, 7H), 0.95 (m, 1H).
General procedure 2. Starting material: compound 79. 1H-NMR (DMSO-d6) δ 10.53 (bs, 1H), 9.32 (bs, 1H), 7.12 (m, 2H), 7.03 (d, 1H), 6.69 (m, 2H), 2.28 (m, 1H), 2.22 (d, 3H), 2.13 (d, 3H), 1.60 (m, 3H), 1.39 (m, 2H), 1.16 (m, 3H), 0.98 (m, 1H), 0.74 (m, 1H).
General procedure 2. Starting material: compound 80. 1H-NMR (DMSO-d6) δ 10.50 (bs, 1H), 9.32 (s, 1H), 7.09 (m, 2H), 7.00 (d, 1H), 6.69 (m, 2H), 2.46 (m, 1H), 2.20 (d, 3H), 2.13 (d, 3H), 1.75-1.15 (m, 11H), 0.85 (m, 1H).
General procedure 2. Starting material: compound 81. 1H-NMR (MeOD) δ 7.28 (d, 1H), 7.17 (d, 1H), 7.07 (m, 2H), 2.98 (m, 1H), 2.29 (d, 3H), 1.7-1.1 (m, 7H), 0.90 (m, 1H).
General procedure 2. Starting material: compound 82. 1H-NMR (DMSO-d6) δ 10.79 (bs, 1H), 9.39 (bs, 1H), 7.37 (m, 2H), 7.13 (m, 2H), 6.71 (m, 2H), 2.37 (m, 1H), 2.32 (d, 3H), 1.58 (m, 3H), 1.42 (m, 2H), 1.2 (m, 3H), 1.01 (m, 1H), 0.75 (m, 1H).
General procedure 2. Starting material: compound 83. 1H-NMR (MeOD) δ 7.42 (d, 1H), 7.31 (d, 1H), 7.15 (m, 2H), 6.74 (m, 2H), 2.65 (m, 1H), 2.38 (d, 3H), 1.8-1.2 (m, 11H), 0.88 (m, 1H).
General procedure 2. Starting material: compound 84. 1H-NMR (DMSO-d6) δ 10.22 (bs, 1H), 9.28 (bs, 1H), 7.13 (m, 2H), 6.69 (m, 3H), 3.69 (s, 3H), 2.96 (m, 1H), 2.15 (s, 3H), 2.08 (s, 3H), 1.65-1.1 (m, 7H), 0.96 (m, 1H).
General procedure 2. Starting material: compound 85. 1H-NMR (DMSO-d6) δ 10.19 (bs, 1H), 9.29 (bs, 1H), 7.14 (m, 2H), 6.70 (m, 3), 3.32 (s, 3H), 2.31 (m, 1H), 2.13 (s, 3H), 2.09 (s, 3H), 1.7-1.1 (m, 8H), 0.98 (m, 1H), 0.76 (m, 1H).
General procedure 2. Starting material: compound 86. 1H-NMR (DMSO-d6) δ 10.16 (bs, 1H), 9.28 (bs, 1H), 7.11 (m, 2H), 6.69 (m, 3H), 3.72 (s, 3H), 2.46 (m, 1H), 2.13 (s, 3H), 2.09 (s, 3H), 1.7-1.1 (m, 11H), 0.88 (m, 1H).
General procedure 2. Starting material: compound 45. 1H-NMR (DMSO-d6) δ 10.35 (bs, 1H), 9.29 (bs, 1H), 7.11 (m, 2H), 7.05 (d, 1H), 6.67 (m, 2H), 6.61 (d, 1H), 3.79 (s, 3H), 2.45 (m, 1H), 2.05 (s, 3H), 1.7-1.2 (m, 11H), 0.83 (m, 1H).
MCF-7 cells were seeded in 96-well plates at 3×103 cells/well in 100 μL of culture medium, 8 wells were left empty for media only controls.
After 24 h the compound titrations were performed, in a separate dilution plate, by serially diluting the compounds of general formula (I) in culture medium. A 100 μL of each dilution was added to the plated cells, this was done in triplicate, and controls (e.g. DMSO and blanks) were included. The plates were incubated for 24 h at 37° C. in a CO2 incubator. The compound titrations were repeated in a separate dilution plate after 24 h. The media plus compound from the assay plates were then aspirated. A 100 μL of media was then added to all wells, followed by 100 μL of each compound dilution. The plates were incubated for a further 48 h at 37° C. in a CO2 incubator (total incubation time 72 h). The number of viable cells was then assessed using Cell Proliferation Reagent WST-1. 10 μL of WST-1 reagent added to each well and incubated for one to four hours at 37° C. in CO2 incubator. The absorbance was measured (450 nm/690 nm).
The activity of compounds of general formula (I) in reducing the number of viable cells was calculated as:
% activity=[(Sc−B)/(So−B)]×100
Sc denotes signal measured in the presence of test compound, So denotes signal detected in the absence of compound, and B denotes background signal, measured in blank wells containing medium only. Analyse data using GraphPad Prism.
Results can be seen in Table 1.
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/EP08/54990 | 4/28/2008 | WO | 00 | 4/16/2010 |
| Number | Date | Country | |
|---|---|---|---|
| 60913625 | Apr 2007 | US |
