CA2091058C - Methylen-oxindole derivatives and process for their preparation - Google Patents
Methylen-oxindole derivatives and process for their preparation Download PDFInfo
- Publication number
- CA2091058C CA2091058C CA002091058A CA2091058A CA2091058C CA 2091058 C CA2091058 C CA 2091058C CA 002091058 A CA002091058 A CA 002091058A CA 2091058 A CA2091058 A CA 2091058A CA 2091058 C CA2091058 C CA 2091058C
- Authority
- CA
- Canada
- Prior art keywords
- hydrogen
- oxindole
- compound
- indolyl
- methylene
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/30—Indoles; Hydrogenated indoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to carbon atoms of the hetero ring
- C07D209/32—Oxygen atoms
- C07D209/34—Oxygen atoms in position 2
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/08—Vasodilators for multiple indications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
Abstract
The invention provides new methylen-indole derivatives of formula (I), wherein R is a group (a) in which R4 is hydrogen, hydroxy, C1-C6 alkoxy, C2-C6 alkanoyloxy, carboxy, nitro or NHR7, wherein R7 is hydrogen or C1-C6 alkyl; R5 is hydrogen, C1-C6 alkyl or halogen; and R6 is hydrogen or C1-C6 alkyl; n is zero, 1 or 2;
R1 is hydrogen, C1-C6 alkyl or C2-C6 alkanoyl;
R2 is hydrogen, C1-C6 alkyl, halogen, cyano, carboxyl, nitro or -NHR7 in which R7 is as defined above; R3 is hydrogen, C1-C6 alkyl or C2-C6 alkanoyl; and the pharmaceutically acceptable salts thereof; and wherein, when, at the same time, R2 is hydrogen, C1-C6 alkyl, halogen or cyano and R3 is hydrogen, R1 and n being as defined above, then at least one of R4, R5 and R6 is other than hydrogen, which are useful as tyrosine kinase inhibitors.
R1 is hydrogen, C1-C6 alkyl or C2-C6 alkanoyl;
R2 is hydrogen, C1-C6 alkyl, halogen, cyano, carboxyl, nitro or -NHR7 in which R7 is as defined above; R3 is hydrogen, C1-C6 alkyl or C2-C6 alkanoyl; and the pharmaceutically acceptable salts thereof; and wherein, when, at the same time, R2 is hydrogen, C1-C6 alkyl, halogen or cyano and R3 is hydrogen, R1 and n being as defined above, then at least one of R4, R5 and R6 is other than hydrogen, which are useful as tyrosine kinase inhibitors.
Description
Methylen-oxindole derivatives and process for their prepara-tion.
The present invention relates to new 3-methylene-2-oxindole derivatives, to a proce~;s Z."or their preparation, to pharma-ceutical compositions containing them and to their use as .
therapeutic agents.
The present invention L:~rovides compounds having the following general formula (I) Rs R
CH
I
R6 I~_ in which R4 is hydrogen, hydroxy, C1~-C6 alkoxy, C2-C6 a3k anoyloxy, carboxy, vitro or NHR,~, wherein R~ is hydrogen or C1-C6 alkyl;
R5 is hydrogen, C1-C6 alkyl or halogen; and R6 is hydrogen or C1-Cf~ alkyl;
n is zero, 1 or 2;
R1 is hydrog~°_n, C1-C6 alkyl or C2--C6 alkanoyl;
R2 is hydrogen, C1-C6 alkyl, halogen, cyano, carboxyl, nitro or -NHR~ :in which R,~ is as defined above;
R3 is hydrogen, C1-C6 alk_,rl or C2-C6 alkanoyl; and the pharmaceutically acceptable salts thereof; and wherein, (l) when, at. the samea tune, R~ is hydrogen, C1-C6 alkyl, halogen or c~~ano and :ft; is hydrogen, R1 and n being as def irJed above , then at. heast. one of R, , RS and R6 is other than hydrogen; and ', ( l l ) when r~ :l s zero and R= and Rk are each hydrogen or CH3, then at. least on~:~ of R2, R9 alld R5 is other than hydrogen.
The inventio» includes within its scope all the possible iso-mers, stereoisomers, in particular Z and E isomers and their mixtures, ane the metabolites arid the metabolic precursors or bio-precursors (otherw:ise knowrv a:~ pro-drugs] of the compounds of formula 1 I ) .
When n is 2, each of the ~-OR1 groups may be the same or dif-ferent.
A substituent -OR1 is p re:ferably linked to position 4, 5, 6 or 7, in particular to position 5 or 7.
The substituent R2 is preferably on the benzene ring moiety, '' "~ 93101182 ~CT/~P92/OI569 in particular linked to position 5.
Of course only one of the substituents R, -OR1 and R2 can be linked to the same position in the indole ring system.
The substituent R4 is preferably linked to position G or S, in particular to position 5.
When R4 is carboxyl, vitro or -NHR~, in which R~ is as de-fined above, the R2 substituent preferably has not the same meanings. Vice versa, when R2 is carboxyl, vitro or -NHR7, in which R,~ is as defined above, the R$ substituent preferably is other than carboxy, vitro or -NHR~.
The alkyl groups, and the alkyl moiety in the alkanoyl groups, may be branched or straight alkyl chain. A C1-C6 alkyl group is preferably a C1-C4 alkyl group, e.g. methyl,, ethyl, propyl, isopropyl, butyl, sec-butyl or tart-butyl, in particular methyl or ethyl. A C2-C6 alkanoyl group is preferably a C2-C4 alkanoyl group, in particular acetyl, prapi~nyl or butyryl.
A halogen is, preferably,.chlorine, bromine or fluorine, in particular bromine.
Pharmaceutically acceptable salts of the compounds of the in-vention include acid addition salts, with inorganic, e.g.
nitric, hydrochloric, hydrobromic, sulphuric, perchloric and phosphoric acids, ar organic, e.g. acetic, propionic, glyc-olic, lactic, oxalic, malonic, malic, maleic, tartaric, citric, benzoic, cinnamic, mandelic and salicylic acids, and salts with inorganic, e.g. alkali metal, especially sodium or po-!VY~ 93/01 1g2 . , PCf/EP92/010"~'' 209.~0~~
tassium, bases or alkaline-earth metal, especially calcium or magnesium bases, or with organic bases, e.g. alkylamines, preferably triethyl-amine.
As stated above the present invention also includes within its scope pharmaceutically acceptable bio-precursors (other-wise known as pro-drugs) of the compounds of formula (I), i.e. compounds which have a different formula to formula (I) above but which nevertheless upon administration to a human being are converted directly or indirectly in vivo into a compound of formula (I). Preferred compounds of the inven-tion are the compounds of formula (I), wherein R is as defined above, R4 is hydroxy, amino, vitro or carboxy and RS and R6 are hydrogen;
R1 is hydrogen or Cl-C6 alkyl;
n is zero or 1;
R2 is hydrogen, carboxy, amino or vitro;
R3 is hydrogen; and the pharmaceutically acceptable salts thereaf.
More preferred compounds of the invention are the compounds 2n of formula (I) in which R is as defined above, R~ is hydroxy, amino or carboxy; and R5 and Ft6 are hydrogen; n is 0 or 1; R1 is hydrogen; R2 is hydrogen, amino or carboxy; R3 is hydrogen; and the.pharma-ceutically acceptable salts thereof.
25~ Examples of specific compounds of the invention are the fol-lowing compounds which, when appropriate, may be either Z-or E-diastereomers or Z,E-mixtures of said diastereomers:
5-hydroxy-3-~.(3'-indolyl)methyleneJ-2-oxindole;
3-((5'-carboxy-3°-ind<:~l;yl )methylene]-2-oxindole;
3- (( S'-amino-~3'-indo13~~1 )methylene]-2-oxindole;
S-carboxy-3-(-(3'-indolyl)methylene]-2-oxindole; y S-amino-3-[(;:1'-indolyl )methylene]-2-oxindole;
5-hydroxy-3-(.(5'-hydrcv,x;v-3'-indolyl )methylene]-2-oxindole;
5-hydroxy-3-((7'-hydrox;~-3'-indolyl)methylene]=~-oxindole; -5-amino-3-[(5'-hydroxy-3'-indolyl)methylene]-c-oxindole;
5-hydroxy-3-(.(5'-amines-~'-indolyl)methylene~-2-oxindole;
5-carboxy-a- (_( 5' -hydrc~xy--3' - indo lyl )me thyl ene] -2-oxindole: ;
5-hydroxy-3-(.(5'-carbc:~xy-3"-indolyl)methylenel-2-oxindole;
5-amino-3-[(~"-hydroxy-3'-indolyl)methylene]-2-oxindole;
~-carboxy-3-(.(7'-hydrax,y-3'-indolyl)methylene]-2-oxindole;
5-metho:cy-3-I.iS'-meth<-~xy-3'-indolyl)-methylene-2-oxindole;
5-acetoxy-J- [ ( 5' Gcetox >>--3 ' ~-indclyl ) -methyienc) -2-oxi.ndole; and 3-[ (5'-vitro--3'-indoly:i:?methylene)~-2-oxindolE
and, if the case, the plus~rmaceutically acceptable salts there-of.
The compounds of the::in~~rentior~, and the pharmaceutically ac-ceptable salts thereof, ca_n be obtained 'by a process compris-ing the condensation of a.n aldehyde of formula (II) I y~-Rz 0 ~ I _'w~-. ( 0 R ~ ; n (-I I ) N r I
R:
WO 93/01182 ~ ~ ~ ~ ~ ~ PCT/EP92/01~'"'~
wherein R1, R2, R3 and n are as defined above, with a compound of formula (III) R (III) wherein Ra, R5 and R6 are as defined above; and, if desired, convert-ing a compound of formula (I) into another compound of formu-la (I), and/or, if desired, converting a compound of formula (I) into a pharmaceutically acceptable salt thereof, andlor, if desired, converting a salt into a free compound, and/or, if desire , separating a mixture of isomers of a compound of for-mula ( i ) into the single isomers. Each of the substituents R2, -CRl and -CHO in a compound of formula ( II ) may be in dependently on either of the.benzene or pyrrole moiety of the indole ring.
The reaetion of a compound of formula (II) with a compound of formula (III) is an analogy process which can be carried out according to known methods, as herebelow described; prefer-ably in the presence of a basic catalyst, e.g. pyridine, pi-peridine, dimethyl~mine, or. a suitable alkali metal hydroxide or alkoxide.
For example the reaction of a compound of formula (II) with ._.,~ 93/oaagZ ~ 0 0 ~. 0 5 8 Pcr/~~2/oas69 a eompound of formula (III) may be carried out under the con-ditions of the Knoevenagel reactions as described e.g. by G.
Jones in Organic Reactions 15, 204 (1967). Suitable catalyst are organic bases such as pyridine, piperidine or diethylamine.
The condensation may be performed in an inert organic solvent e.g. pyridine, ethanol, methanol, benzene or dioxane at tem-peratures ranging from about 0°C to about 100°C.
Preferably the reaction is carried out in warm ethanol solu-tion in the presence of piperidine catalyst.
A compound of formula (I) can be converted into another com-pound of formula (I) according to known methods. For example the de-etherification of a compound of formula (I), wherein one or more R1 substituents are C1-C6 alkyl, so as to obtain a compound of formula (I) wherein one or more R1 substituents are hydrogen may be performed by well known methods in organic chemistry. In the case of a phenolic methyl ether the cleavage can be carried out for example with boron tribromide as des-cribed by J.F.N. McOmie in Tetrahedron 24, 2289 (1968). It is advisable to use about 1 mole of boron tribromide for each ether group together with an extra mol of reagent for each group containing a potentially basic nitrogen or oxygen. The reaction may be performed in an inert organic solvent such as methylene chloride, pentane or benzene under an inert, e.g.
nitrogen, atmosphere at temperatures ranging from about -78°C
to about room tempe~ratuse.
22551-9b _g_ The acylation of a compound of formula () wherein one or more of -ORl and/or R~~ is hydroxy, so as to obtain a corres-ponding compound of formula ( I ) wherein one or more of -0R1 and/or R4 is a C2-C6 alkanoyloxy grbup, may be obtained by reaction with a reactive derivative of a suitable carboxylic acid, such as an anh;,~dride orv halide, in the presence of a basic agent, at temperatures ranging from about 0°C to about 50°C.
Preferably the acylaticn is carried out by reaction with the respective anh!~dride in 'the presence of an organic base, such as pyridine.
The optional salification of a compound of formula (I) as well as the.convers:ion of a salt into the free compound and the se-paration of a rnixture ef isomers into the single isomers may be carried out by conventional methods.
For example the= separation of a mixture of geometric isomers, e.g. cis- and traps-isomers, may be carried out by fractional crystallization from a suitable solvent or by chromatography, either column chromatography or high pressure liquid chromato-graphy.
The compounds of formula (II) may be obtained according to known methods from compounds of formula (IV) R~
. _ {OR) )n ~N ..~::
~3 -w 9~/Ol lsx 2 D 9 '10 ~5 $
_ g _ For example the 3-formylindole derivatives of formula (II) can be obtained from a compound (IV) by formylation with N-methylformanilide and phosphorous oxychloride according to the well known Vilsmeyer-Haack method (for a review see W.G.
Jaokson et al., J.Am.Chem.Soc. 1981, 103, 533). The 2-formylindole derivatives are obtained when the 3-position is occupied.
The compounds of formula (III) and (IV) are known or may be obtained by known methods from known compounds.
PHARMACOLOGY
The compounds of the invention possess specific tyrosine kinase inhibiting activity. Hence they can be useful in the treatment of cancer and other pathological proliferative conditions, such as to inhibit the development of the atheromatous plaque, in mammals, including humans.
Typical therapeutical indications according to.the latter use are reocclusion following coronary angioplasty and in general decrease of coronary artery disease.
Recent studies on the molecular basis of neoplastic transformation have identified a family of genes, designed oncogenes, whose aberrant expression causes tumorigenesis.
For example, the RNA tumor viruses possess such an oncogene sequence whose expression determines neoplastic conversion of infected cells. Several of their oncogene-encoded proteins, SUCK as pp60"'-'~°, p70°~o-xa.r p130'~~o-sn. and P70°a~-sae display protein tyrosine kinase activity, that is they catalyse the transfer of the ~-phosphate from adenosine - triphosphate (ATP) to tyrosine residues in protein substrate.
In normal cells, several growth factor receptors, for example the receptors, for PDGF,:EGE, d-TGF and. insulin, display .. tyrosine kinase.activity. -?~~~~~. ~~$
wo 93iousz ~crm~z/oiG~., - to -Binding pf the growth factor (GF) activates the receptor tyrosine kinase to undergo autophosphorylation and to phosphorylate closely adjacent molecules on tyrosine.
Therefore, it is thought that the phosphorylation of these tyrosine kinase receptors plays an important role in signal transduction and that the principal function of tyrosine kinase activity in normal cells is to regulate cell growth.
Perturbation of this activity by oncogenic tyrosine kinases that are either overproduced and/or display altered substrate specificity may cause loss of growth control and/or neoplastic transformation. Accordingly, a specific inhibitor of tyrosine kinases can be useful in investigating the mechanism of carcinogenesis, cell proliferation and differentiations and it can be effective in prevention and chemotherapy of cancer and in other pathological proliferative conditions.
The tyrosine specific protein kinase activity of these compounds is shown by the in vitro tests described herebelow.
v-abl kinase purification The enzyme used in the Abelson tyrosine kinase p95°'~''~ which was produced and isolated as follows: 1 Liter cultures of HB-130 cells in LB medium, supplemented with ampicillin, were grown at 30°C as described by Wang et al, in J.Biol.Chem.2_60, 64 (1985). The expression of v-abl protein was induced by shifting the temperature to 42°C for 3-4 h. The bacterial cells were collected on ice, centrifuged and frozen in liquid nitrogen. The cell pellet was lysed as described by Ferguson et al. in J.Biol.Chem. 260, 3652 (1985) and in Biochem.J.
257, 321 (1989). Briefly, bacterial proteins were removed by differential ~solubilization with sodium deoxycholate, NaCl and ~-octylglucopyranoside..in different steps. Insoluble v-abl protein was solubilized with 2M FCSCN and dialyzed against 100 volumes of 50 mM Tris-HC1, pH 7.5, 1mM EDTA and 0.1 mM
- .L 1 -dithiothreitol (buffer A). The soluble proteins were separated by chromatography on a grepacked Mono-QTM anion exchange column in a f.p.l.c. system. The v-abl activity was eluted with a linear KC1 gradient (O-I M buffer A). Active fractions were pooled, made 50% in glycerol and stored in small aliquots at -20°C.
Myelin Basic Proteins phophor~lation assay The in vitro test with Myelin Basic Protein as substrate was carried out as follows; Protein phasphorylation was performed by incubating. ~0 ng of purified v-abl kinase, 1.5 pCi ~[~ 3~P]
ATP, 10 EzM cold ATP, 56 pM myelin basic protein in 50 ~1 of Tris-HC1 25 mM, pH 8.0, containing MgClz 10 mM, dithiothrei-tol 0.1 mM (kinase buffer) at 22°C.. The reaction was stopped by addition of equal ~rolumes of 2-fold concentrated Laem~nli electrophoresis buffer [U. K. Laem~nli, Nature (London) 230, 680 (1970). Samples were boiled again for 3 min and separated in SDS-PAGE (15% acrylamide). Gels were dried and exposed to autoradiographic films for 15-30 min at -70°C. Bands were located by autoradiography, excised from gel and counted in a liquid scintillation counter.
Autophosphorylation assay For the autophosphorylation assay the v-abl kinase was immunoprecipitated With antiphosphotyrosine antibodies and the resulting immunocomplex analyzed in 50 ~l of kinase buffer in the presence of 10 ~ZM ATP and 10 ~Ci [ ~-3~P]-ATP.
The reaction was stopped after 15 min at room temperature with boiling Laemmli buffer. Samples were boiled again for 3 min and separated in SDS-PAGE (8% acrylamide). Gels were dried and exposed to autoradiographic films for up to 3 h at -70°C. Bands were located by autoradiography, excised and counted as above.
W093/01182 z~~d:~o~s PCTlEP92/015f - i~ -In table I and II a representative compound of this invention is compared with the corresponding non hydroxylated analog which is encompassed by the general formula of patent ,.
application w~91/13055. The comparison shows that the introduction of an hydroxyl graup, while increasing only .
slightly the potency toward exogenous substrates significantly enhances the inhibitory activity on the autophosphorylation.
Table I. Myelin Pasic Protein phosphorylation assay.
ICso(~M) 5-hydroxy-3-j(3'-indolyl)methylen]-2-oxindole 0.4 3-[(3'-indolyl)methylen]-2-oxindole 0.6 Table II. Autophosphorylation assay.
ICSO(NM) 5-hydroxy-3-j(3'-indolyl)methylen]-2-oxindole 0.4 3-j(3'-indolyl)methylen]-2-oxindole : 10 In view of their high activity and low toxicity, the compounds of the invention can be used safely in medi-cine. For example, the approximate acute toxicity (LDP) of the compounds of the invention in the mouse, deter-mined by single administration of increasing doses and measured on the seventh day after the treatment was found to be negligible.
The compounds of the invention can be administered in a variety of dosage forms, e.g. orally, in the form of tablets, capsules, sugar or film-coated tablets, liquid solutions or suspensions; rectally, in the form of sup-positories; parenterally, e.g. intramuscularly, or by intravenous injection or infusion; or topically.
The dosage depends on the age, weight, conditions of the patient and administration route; for example, the dosage adopted for oral administration to adult humans may range from about 10 to about 150-200 mg per dose, from 1 to 5 times daily. Of course, these dosage regimens may be ad-justed to provide the optimal therapeutic response.
The invention includes pharmaceutical compositions com-prising a compound of formula (I) or a pharmaceutically acceptable salt thereof in association with a pharmaceu-tically acceptable excipient (which can be a carrier or diluent).
The invention also provides a pharmaceutical composition comprising a suitable carrier and/or diluent and, as an active principle, a compound selected from 3-[(1'-methyl-3'-indolyl)methylen]-2-oxindole, 3-[(3'-indolyl)methylen]-1-methyl-2-oxindole, and the pharmaceutically acceptable salts thereof, the compound or salt being either the Z- or E-diastereoisomer or a Z/E
mixture of diastereoisomers.
The compounds defined above as an active principle of the pharmaceutical composition are also provided by the invention for use as a tyrosine kinase inhibitor, as an anti-proliferative agent, as an anti-cancer agent or in the treatment of coronary a2tery disease.
The pharmaceutical compositions containing'the compounds of the invention are usually prepared following conven-tional methods and are administered in a pharmaceutical-ly suitable form.
For example, the solid oral forms may contain, together with the active compound, diluents, e.g., lactose, dex-trose, saccharose, cellulose, corn starch or potato starch; lubrican~.s, e.g. silica, talc, stearic acid, magnesium or calcium stearate, and/or polyethylene glyc-ols; binding agents, e.g. starches, arabic gums, gelatin, methylcellulose, carboxymethylcellulose or polyvinyl pyr-rolidone; disaggregating agents, e.g. a starch, alginic acid, alginates or sodium starch glycolate, effervescing mixtures; dyestuffs; sweeteners; wetting agents, such as ,v lecithin, polysorbates, laurylsulphates; and, in general, non-toxic and pharmacologically inactive substances used in pharmaceutical formulations. Said pharmaceutical pre-parations may be manufactured in known manner, for exacr~le, by means of mixing, granulating, tabletting, sugar-coat-ing or film-coating processes.
"'~ 93/01182 ~ ~ ~ ~ ~ ~ ~ PCTJEP'92/01569 The liquid dispersion for oral administration may be, e.g., syrups, emulsions and suspensions.
The syrup may contain as carrier, for example, sacchar-ose or saccharose with glycerine and/or mannitol and/or sorbitol.
The suspensions and the emulsions may contain as carrier, for example, a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl al-cohol.
The suspensions or solutions for intramuscular injections may contain, together with the active comnnmn~_ a r,h~.._ maceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oleate, glycols, e.g. propylene glyeol, and, if desired, a suitable amount of lidocaine hydro-chloride.
The solutions for intravenous injections or infusion may contain as carrier, for example; sterile water or, pre-ferably, they may be in the form of sterile, aqueous, isotonic saline solutions.
The suppositories may contain, together with the active compound, a pharmaceutically acceptable carrier, e.g.
cocoa-butter, polyethylene glycol, a palyoxyethylene sorbitan fatty acid ester surfactant or lecithin.
Compositions gor topical application, e.g. creaaas, lo-2S Lions, or pastes, can be prepared by admixing the active WO 93/01182 2 ~ ~ ~ ~ l~ ~ PCf/EP92/Ol~~'°' ingredient with a conventional oleaginous or emulsifying excipient.
A further object of the present invention is a combined method of treatment of cancer in mammals, including humans, in need of such treatment, said method compris-ing administering 1) a compound of formula (I), or a pharmaceutically ac-ceptable salt thereof, and 2) an additional antitumor agent, in amounts and close enough together in time sufficient to produce a the-rapeutically useful effect.
Object of the present invention is also to provide pro-ducts containing a compound of formula (I), or a pharma-ceutically acceptable salt, and an additional antitumor agent as a combined preparation for simultaneous, separ-ate or sequential use in anti-cancer therapy.
The term "antitumor agent" is meant to comprise both a single antltumor drug and °'cocktails" i.e. a mixture of such drugs, according to the clinical practice.
An titvanor agents that can be foranulated with a compound of the invention or, alternatively, can be administered in a combined method of treatment, are e.g. doxorubicin, daunomycin, eplrubicin, idarubicin, etoposide, flucro-uracil, mephalan, cyclophospha~aide, bleonycin, vinblas-~25 tin arid mitomycin or a mixture of two or snore thereof.
"'O 93/01182 1'~t'T/EP92/01569 7 _ T2-~e vompcundS of the invention can therefore be used in a treatment to ameliorate a cancer. They may be administered to~ a patient suffering from a cancer treatable with an antitumor agent, for example an anthracycline glycoside such as doxorubicin, daunomycin, epirubicin or idarubicin as mentioned above, together with the antiproliferative agent.
A compound of the invention and an antitumor agent such as an anthracycline glycoside can be administered to improve the ZO condition of a patient having a leukaemia such as myeloblastic leukaemia, lymphoma, sarcoma, neuroblastoma, wilm°s tumor or malignant neoplasm of the bladder, breast, lung or thyroid.
The following ekamples illustrate but do not limit the invention.
WO 93/01182 ~ ~ ~ ~ ~ Q8 - 1'GT/1rP92/Ol~'-'' Ex amp 1 a 1 5-Hydroxy-3-[(3'-indolyl)methylene]-2-oxindole ~I, ~R as defined, n = 0, R2 = R3 = R5 = R6 = H, R4 = 5-OH]
A solution of 3-indolecarboxaldehyde (145 mg, 1 mmol), 5-hydroxy-2-oxindole (149 mg, 1 mmol) and piperidine (60 mg, 0.7 mmol) in absolute ethanol (10 ml) is heated for 3 h at 60°C under nitrogen. Then the reaction mixture is chilled and evaporated under vacuum to dryness. The residue is submitted to column chromatography over silica gel using methylene-chloride/ethanol 4 % as eluant. Pure title compound is so ob-tained in 60 % yield (166 mg). - Alternatively, the reaction mixture is concentrated under vacuum and then chilled to 0-5°C, the precipitate filtered, the residue wash ed With ice-cooled ethanol and finally dried under vacuum. Compounds of higher purity are obtained by further crystallization from ethanol.
C17H12N202 requires: C 73.89 H 4.38 N 10.14 found: C 73.51 H 4.21 N 9.92 MS m/z: 276 IR cm 1 (7CBr): 3600-2500 (NH, 0H), 1650 (CO), 1600, 1580, 1530, 1480 (C=C) m.p. 293°C (dec.).
According to the above described procedure, the following compounds can be prepared:
5-carboxy-3-t(3'-indolyl)methylenel-2-oxindole;
5-amino-3-[3'-indolyl)methylene~-2-oxindole;
'"O 93/f1182 - 19 - PCT/EP92/O1S69 5-hydroxy-3-[(5'hydroxy-3'-indolyl)methylene]-2-oxindole;
NfS m/2 : ~ 282 IR cm-1 (KBr); 3600-2600 (NH, OH), 1655 (CO), 1605, 1585, 1535 (C=C) 5-hydroxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
3-L(5a7'-dihydroxy-3'-indolyl)methylene]-2-oxindole;
5-amino-3-C(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-amino-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-~(5'-carboxy-3'-indolyl)methylene]-2-oxindole;
5-amina-3-((7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-methoxy-3-((5'-tr~ethoxy-3'-indolyl)-methYlene'-2-oxindole;
5-acetoxy-3-~(5'-acetoxy-3'-indolyl)mettrylene,?-2-oxindole;
3-r(5'-carboxy-3'-indolyl)methylene]-2-oxindole, C18H12N203 requires: C 71.04; H 3.98; N 9.21 -found : C 70.83; H 4.6; N 8.85 MS m/z : 304;
m.p. > 330°dec.
IR cm 1 (KBr): 3600-3000 (NH), 3000-2100 (OH), 1710 (CO), 1640, 1620, 1600, 1550 Carom);
3-t(5'-amino-3°-indolyl)methylene~ -2-oxindole, C17H13N30 requires : C 74.14; H 4.71; N 15.26 ' found : C 73.88; H 4.51; N 14.91 2S MS m/z 275;
m.p. 250°dec.
IR cm 1 (KBr) . 3300, 2380 (NH), 1670 (CO), 1600, 1510 (C=C);
WO 93/o118z 2 ~ (~ ~ ~ ~ ~ Pc~r~EP9z,/ol~, '.
3-[(5'-nitro-3'-indolyl)methylen~ -2-oxindole, C17Hi1N303 requires : C 66.88; H 3.63; N 13.76 found : C 66.58; H 3.74; N 13.64 MS m/z 305;
m.p. > 350°C
IR cm-1 (KBr): 3350, 3230 (NH), 1680 (CO), 1620, 1605, 1580 (c=c), 1530, 1340 (N02);
3- ~(1'-methyl-3°-indolyl)methylen] -2-oxindole, C18H14N20 requires : C 78.81; H 5. I4; N 10.21 found : C 78.42; H 5.17; N 10.00 MS m/z 274 m.p. 230'C (dec.) IR cm 1 (KRr) . 3300--2000 (NH), 1680 (C0), 1610, 1600, 1570, 1500 (C=C);
3- [(3'-indolyl)methylen~-1-methyl-2-oxindole, C18H14N20 requires : C 78.81; N 5.14; N 10.21 found : C 78.61; H 5.16; N 10.23 MS m/z 274 m.p. 274°C
IR cm-1 (KBr) . 3220 (NH), 1675 (CO), 1605, 1500, 1490 (C=C).
93/01182 - 21 - . 2. ~ ~ ~ ~ ~ ~ P~.'T/EP92/01569 Example 2 5-Hydroxy-3-[(5'-hydroxy-3'-indolyl)methylene~-2-oxindole rI, R as defined, n = 1, R1 = R2 = R3 = R5 = R6 = H, R4=5-OH' The starting material for this de-etherification example is 5-methoxy-3- [(5'-methoxy-3'-indolyl)methylene~ -2-oxindole, which can be obtained according to the procedure described in Ex~ple 1.
To a stirred solution of 5-methoxy-3- C(5'-methoxy-3'-indolyl) methylene]-2-oxindole (310 mg, 1 mmol) in anhydrous dichloro-methane (10 ml) is added at -78°C under nitrogen, over a pe-riod of 10 min, a 1.0 M solution of boron tribromide in di-chloromethane (3 ml, 3 mmol). The resulting mixture is stir-red for another 1 h at -78°C and then allowed to warm to room temperature. After stirring for 1.5 h at 20-25°C. the mixture is cooled to -10°C and then quenched by the dropwise addition of water (10 ml) over a 10-min period. After addition of ethylacetate the organic layer is separated, washed with water, dried with Na2S04 and evaporated under vaeuum to dry-ness. The residue is crystallized from ethanol thus giving 198 mg of pure title compound (yield 70 %).
C17H12N203 requires: C 72.33 H 4.29 N 6.38 found: C 72.11 H 4.07 N 6.29 MS m/2: 282 IR cm 1 (KBr): 3600-2600 (NH, OH), 1655 (C0), 25' 1605, 1585, 1535 (C=C) , According to, the above descx°ibed procedure and starting from the corresponding methyletlaer, the. hydroxyl compounds men-tioned in Example 1 can be obtained.v wc~ 9momsz '~,' ~ ~ ~. ~ ~ $ ~~crm~ziol~ ._:_, Example 3 5-Acetoxy-3- ~(5'-acetoxy-3'-indolyl)methylene~-2-oxindole ~I, R as defined, n = 1, R1 = Ac, R4 = 5-OAc, R2=R3=R5=R6=H~
The starting material for this acylation example is 5-hydroxy-3-[(5'-hydroxy-3'-indolyl)methylene~ -2-oxindole, which may be obtained according to the procedure described in Examples 1 an d 2 .
To a cooled solution of 5-hydroxy-3-[(5'-hydroxy-3'-indolyl) methylene'-2-oxindole (282 mg, 1 mmol) in dry pyridine (0.5 ml) is added acetic anhydride (306 mg, 3 mmol) and the.mix-ture maintained at 0-5°C overnight. Thereupon the mixture concentrated under vacuum, the residue dissolved in dichloro-methane, the organic layer washed with water and then evapor-ated under reduced pressure. The crude product is crystallized from chlaroform/methanol to yield pure title compound in 80%
yield (301 mg).
C21H16N205 requires: C 67:02 H 4.29 N 7.44 found: C 66.91 H 4.05 N 7.29 MS m/z: 376 IR cm 1 (KRr): 3600-3200 (NH), 1750 (CH3C00), 1650 (CONH), 1600, 1580, 1530 (C=C) According to the above described procedure, the hydroxyl com-pounds obtained in~Example l and 2 can be transformed into the eorrespondfng.~~CZ-C6 alkanoyloxyd~rivativese .
7 93/01182 '~ ~ ~ ~ fl ~ ~ PC.'1'1EP92/01569 Example 4 5-Nitro-3-indolealdehyde ~I, n -_ 0, R2 = 5-N02, R3 = H;
A mixture of N-methylformanilide~(176 mg, 1.3 mmol) and phos-S phorous oxychloride (199 mg, 1.3 mmol) is stirred for 15 min at 20-25°C under nitrogen. then a solution of 5-nitroindole (162 mg, 1 mmol) in 1,2-dichloroethane (S ml) is added and the mixture heated to reflux for 3 h. After cooling the mix-ture is poured onto iced water, the precipitate filtered off 1p and washed with water. Thereupon the residue is chromato-graphed over silica gel using benzene/ethyl acetate as eluant.
Thus pure title compound is obtained in 80 % yield (152 mg).
CgH6N203 requires: C 56.85 H 3.18 N 14.73 found: C 56.79 H 3.01 N 14.51 15 MS m/z: 190 IR cm-1 (KHr): 3140, 3090 (NH), 1650 (CO), 1511, 1345 (N02) These vitro-intermediates besides leading to final products of formula (I) with R 2 = N02 give also rise to final proudcts 20 with R 2 = NH2 obtainable from the former by reduction.
~y proceding analogously, the following protected interme-diates can be prepared, which after deprotection at a suit-able stage of the synthesis give also rise to final product of formula (I) with free carboxyl (R 2 = COON) and free hydr-25 oxyl (R 2 = OH) respectively:
3-carbcx~ethoxy-3-indolealdehyde; and 3-methoxy-3-indolealdehyde.
WO 93J01182 ~ ~ ~ ~ ~~ ~ ~ c..., PCE'/EP92/Oi., _ ~4 _ Example 5 Tablets each weighing 0.150 g and containing 25 mg of the active substance, can be manufactured as follows:
composition (for 10,000 tablets):
5-Hydroxy-3-[(3'-indolyl)methylene3-2-oxindole 250 g Lactose 800 g Corn starch 415 g Talc powder 30 g P:agnesium stearate 5 g Th a 5-hydroxy-3-~(3'-indolyl}methylene~-2-oxindole, the-lactose and half the corn starch are mixed; the mixture is then forc-ed through a sieve of 0.5 mm mesh size.
Corn starch (10 g} is suspended in warm water (90 ml) and the resulting paste is used to granulate the powder. The granul-ate is dried, comminuted on a sieve of 1.4 mm mesh size, then the remaining quantity of starch, talc and magnesium stearate are added, carefully mixed and processed into tablets.
~='d!O 93/0i182 ~fT/EP92/Ole' ~~
Example 6 Capsules, each dosed at 0.200 g and containing 20 mg of the active substance can be prepared.
Composition for 500 capsules:
3-[(5'-amino-3°-indolyl)methylene~ -2-oxindole 10 g Lactose 80 g Corn starch 5 g , Magnesium stearate 5 g This formulation is encapsulated in two-piece hard gelatin capsules and dosed at 0.200 g for each capsule.
The present invention relates to new 3-methylene-2-oxindole derivatives, to a proce~;s Z."or their preparation, to pharma-ceutical compositions containing them and to their use as .
therapeutic agents.
The present invention L:~rovides compounds having the following general formula (I) Rs R
CH
I
R6 I~_ in which R4 is hydrogen, hydroxy, C1~-C6 alkoxy, C2-C6 a3k anoyloxy, carboxy, vitro or NHR,~, wherein R~ is hydrogen or C1-C6 alkyl;
R5 is hydrogen, C1-C6 alkyl or halogen; and R6 is hydrogen or C1-Cf~ alkyl;
n is zero, 1 or 2;
R1 is hydrog~°_n, C1-C6 alkyl or C2--C6 alkanoyl;
R2 is hydrogen, C1-C6 alkyl, halogen, cyano, carboxyl, nitro or -NHR~ :in which R,~ is as defined above;
R3 is hydrogen, C1-C6 alk_,rl or C2-C6 alkanoyl; and the pharmaceutically acceptable salts thereof; and wherein, (l) when, at. the samea tune, R~ is hydrogen, C1-C6 alkyl, halogen or c~~ano and :ft; is hydrogen, R1 and n being as def irJed above , then at. heast. one of R, , RS and R6 is other than hydrogen; and ', ( l l ) when r~ :l s zero and R= and Rk are each hydrogen or CH3, then at. least on~:~ of R2, R9 alld R5 is other than hydrogen.
The inventio» includes within its scope all the possible iso-mers, stereoisomers, in particular Z and E isomers and their mixtures, ane the metabolites arid the metabolic precursors or bio-precursors (otherw:ise knowrv a:~ pro-drugs] of the compounds of formula 1 I ) .
When n is 2, each of the ~-OR1 groups may be the same or dif-ferent.
A substituent -OR1 is p re:ferably linked to position 4, 5, 6 or 7, in particular to position 5 or 7.
The substituent R2 is preferably on the benzene ring moiety, '' "~ 93101182 ~CT/~P92/OI569 in particular linked to position 5.
Of course only one of the substituents R, -OR1 and R2 can be linked to the same position in the indole ring system.
The substituent R4 is preferably linked to position G or S, in particular to position 5.
When R4 is carboxyl, vitro or -NHR~, in which R~ is as de-fined above, the R2 substituent preferably has not the same meanings. Vice versa, when R2 is carboxyl, vitro or -NHR7, in which R,~ is as defined above, the R$ substituent preferably is other than carboxy, vitro or -NHR~.
The alkyl groups, and the alkyl moiety in the alkanoyl groups, may be branched or straight alkyl chain. A C1-C6 alkyl group is preferably a C1-C4 alkyl group, e.g. methyl,, ethyl, propyl, isopropyl, butyl, sec-butyl or tart-butyl, in particular methyl or ethyl. A C2-C6 alkanoyl group is preferably a C2-C4 alkanoyl group, in particular acetyl, prapi~nyl or butyryl.
A halogen is, preferably,.chlorine, bromine or fluorine, in particular bromine.
Pharmaceutically acceptable salts of the compounds of the in-vention include acid addition salts, with inorganic, e.g.
nitric, hydrochloric, hydrobromic, sulphuric, perchloric and phosphoric acids, ar organic, e.g. acetic, propionic, glyc-olic, lactic, oxalic, malonic, malic, maleic, tartaric, citric, benzoic, cinnamic, mandelic and salicylic acids, and salts with inorganic, e.g. alkali metal, especially sodium or po-!VY~ 93/01 1g2 . , PCf/EP92/010"~'' 209.~0~~
tassium, bases or alkaline-earth metal, especially calcium or magnesium bases, or with organic bases, e.g. alkylamines, preferably triethyl-amine.
As stated above the present invention also includes within its scope pharmaceutically acceptable bio-precursors (other-wise known as pro-drugs) of the compounds of formula (I), i.e. compounds which have a different formula to formula (I) above but which nevertheless upon administration to a human being are converted directly or indirectly in vivo into a compound of formula (I). Preferred compounds of the inven-tion are the compounds of formula (I), wherein R is as defined above, R4 is hydroxy, amino, vitro or carboxy and RS and R6 are hydrogen;
R1 is hydrogen or Cl-C6 alkyl;
n is zero or 1;
R2 is hydrogen, carboxy, amino or vitro;
R3 is hydrogen; and the pharmaceutically acceptable salts thereaf.
More preferred compounds of the invention are the compounds 2n of formula (I) in which R is as defined above, R~ is hydroxy, amino or carboxy; and R5 and Ft6 are hydrogen; n is 0 or 1; R1 is hydrogen; R2 is hydrogen, amino or carboxy; R3 is hydrogen; and the.pharma-ceutically acceptable salts thereof.
25~ Examples of specific compounds of the invention are the fol-lowing compounds which, when appropriate, may be either Z-or E-diastereomers or Z,E-mixtures of said diastereomers:
5-hydroxy-3-~.(3'-indolyl)methyleneJ-2-oxindole;
3-((5'-carboxy-3°-ind<:~l;yl )methylene]-2-oxindole;
3- (( S'-amino-~3'-indo13~~1 )methylene]-2-oxindole;
S-carboxy-3-(-(3'-indolyl)methylene]-2-oxindole; y S-amino-3-[(;:1'-indolyl )methylene]-2-oxindole;
5-hydroxy-3-(.(5'-hydrcv,x;v-3'-indolyl )methylene]-2-oxindole;
5-hydroxy-3-((7'-hydrox;~-3'-indolyl)methylene]=~-oxindole; -5-amino-3-[(5'-hydroxy-3'-indolyl)methylene]-c-oxindole;
5-hydroxy-3-(.(5'-amines-~'-indolyl)methylene~-2-oxindole;
5-carboxy-a- (_( 5' -hydrc~xy--3' - indo lyl )me thyl ene] -2-oxindole: ;
5-hydroxy-3-(.(5'-carbc:~xy-3"-indolyl)methylenel-2-oxindole;
5-amino-3-[(~"-hydroxy-3'-indolyl)methylene]-2-oxindole;
~-carboxy-3-(.(7'-hydrax,y-3'-indolyl)methylene]-2-oxindole;
5-metho:cy-3-I.iS'-meth<-~xy-3'-indolyl)-methylene-2-oxindole;
5-acetoxy-J- [ ( 5' Gcetox >>--3 ' ~-indclyl ) -methyienc) -2-oxi.ndole; and 3-[ (5'-vitro--3'-indoly:i:?methylene)~-2-oxindolE
and, if the case, the plus~rmaceutically acceptable salts there-of.
The compounds of the::in~~rentior~, and the pharmaceutically ac-ceptable salts thereof, ca_n be obtained 'by a process compris-ing the condensation of a.n aldehyde of formula (II) I y~-Rz 0 ~ I _'w~-. ( 0 R ~ ; n (-I I ) N r I
R:
WO 93/01182 ~ ~ ~ ~ ~ ~ PCT/EP92/01~'"'~
wherein R1, R2, R3 and n are as defined above, with a compound of formula (III) R (III) wherein Ra, R5 and R6 are as defined above; and, if desired, convert-ing a compound of formula (I) into another compound of formu-la (I), and/or, if desired, converting a compound of formula (I) into a pharmaceutically acceptable salt thereof, andlor, if desired, converting a salt into a free compound, and/or, if desire , separating a mixture of isomers of a compound of for-mula ( i ) into the single isomers. Each of the substituents R2, -CRl and -CHO in a compound of formula ( II ) may be in dependently on either of the.benzene or pyrrole moiety of the indole ring.
The reaetion of a compound of formula (II) with a compound of formula (III) is an analogy process which can be carried out according to known methods, as herebelow described; prefer-ably in the presence of a basic catalyst, e.g. pyridine, pi-peridine, dimethyl~mine, or. a suitable alkali metal hydroxide or alkoxide.
For example the reaction of a compound of formula (II) with ._.,~ 93/oaagZ ~ 0 0 ~. 0 5 8 Pcr/~~2/oas69 a eompound of formula (III) may be carried out under the con-ditions of the Knoevenagel reactions as described e.g. by G.
Jones in Organic Reactions 15, 204 (1967). Suitable catalyst are organic bases such as pyridine, piperidine or diethylamine.
The condensation may be performed in an inert organic solvent e.g. pyridine, ethanol, methanol, benzene or dioxane at tem-peratures ranging from about 0°C to about 100°C.
Preferably the reaction is carried out in warm ethanol solu-tion in the presence of piperidine catalyst.
A compound of formula (I) can be converted into another com-pound of formula (I) according to known methods. For example the de-etherification of a compound of formula (I), wherein one or more R1 substituents are C1-C6 alkyl, so as to obtain a compound of formula (I) wherein one or more R1 substituents are hydrogen may be performed by well known methods in organic chemistry. In the case of a phenolic methyl ether the cleavage can be carried out for example with boron tribromide as des-cribed by J.F.N. McOmie in Tetrahedron 24, 2289 (1968). It is advisable to use about 1 mole of boron tribromide for each ether group together with an extra mol of reagent for each group containing a potentially basic nitrogen or oxygen. The reaction may be performed in an inert organic solvent such as methylene chloride, pentane or benzene under an inert, e.g.
nitrogen, atmosphere at temperatures ranging from about -78°C
to about room tempe~ratuse.
22551-9b _g_ The acylation of a compound of formula () wherein one or more of -ORl and/or R~~ is hydroxy, so as to obtain a corres-ponding compound of formula ( I ) wherein one or more of -0R1 and/or R4 is a C2-C6 alkanoyloxy grbup, may be obtained by reaction with a reactive derivative of a suitable carboxylic acid, such as an anh;,~dride orv halide, in the presence of a basic agent, at temperatures ranging from about 0°C to about 50°C.
Preferably the acylaticn is carried out by reaction with the respective anh!~dride in 'the presence of an organic base, such as pyridine.
The optional salification of a compound of formula (I) as well as the.convers:ion of a salt into the free compound and the se-paration of a rnixture ef isomers into the single isomers may be carried out by conventional methods.
For example the= separation of a mixture of geometric isomers, e.g. cis- and traps-isomers, may be carried out by fractional crystallization from a suitable solvent or by chromatography, either column chromatography or high pressure liquid chromato-graphy.
The compounds of formula (II) may be obtained according to known methods from compounds of formula (IV) R~
. _ {OR) )n ~N ..~::
~3 -w 9~/Ol lsx 2 D 9 '10 ~5 $
_ g _ For example the 3-formylindole derivatives of formula (II) can be obtained from a compound (IV) by formylation with N-methylformanilide and phosphorous oxychloride according to the well known Vilsmeyer-Haack method (for a review see W.G.
Jaokson et al., J.Am.Chem.Soc. 1981, 103, 533). The 2-formylindole derivatives are obtained when the 3-position is occupied.
The compounds of formula (III) and (IV) are known or may be obtained by known methods from known compounds.
PHARMACOLOGY
The compounds of the invention possess specific tyrosine kinase inhibiting activity. Hence they can be useful in the treatment of cancer and other pathological proliferative conditions, such as to inhibit the development of the atheromatous plaque, in mammals, including humans.
Typical therapeutical indications according to.the latter use are reocclusion following coronary angioplasty and in general decrease of coronary artery disease.
Recent studies on the molecular basis of neoplastic transformation have identified a family of genes, designed oncogenes, whose aberrant expression causes tumorigenesis.
For example, the RNA tumor viruses possess such an oncogene sequence whose expression determines neoplastic conversion of infected cells. Several of their oncogene-encoded proteins, SUCK as pp60"'-'~°, p70°~o-xa.r p130'~~o-sn. and P70°a~-sae display protein tyrosine kinase activity, that is they catalyse the transfer of the ~-phosphate from adenosine - triphosphate (ATP) to tyrosine residues in protein substrate.
In normal cells, several growth factor receptors, for example the receptors, for PDGF,:EGE, d-TGF and. insulin, display .. tyrosine kinase.activity. -?~~~~~. ~~$
wo 93iousz ~crm~z/oiG~., - to -Binding pf the growth factor (GF) activates the receptor tyrosine kinase to undergo autophosphorylation and to phosphorylate closely adjacent molecules on tyrosine.
Therefore, it is thought that the phosphorylation of these tyrosine kinase receptors plays an important role in signal transduction and that the principal function of tyrosine kinase activity in normal cells is to regulate cell growth.
Perturbation of this activity by oncogenic tyrosine kinases that are either overproduced and/or display altered substrate specificity may cause loss of growth control and/or neoplastic transformation. Accordingly, a specific inhibitor of tyrosine kinases can be useful in investigating the mechanism of carcinogenesis, cell proliferation and differentiations and it can be effective in prevention and chemotherapy of cancer and in other pathological proliferative conditions.
The tyrosine specific protein kinase activity of these compounds is shown by the in vitro tests described herebelow.
v-abl kinase purification The enzyme used in the Abelson tyrosine kinase p95°'~''~ which was produced and isolated as follows: 1 Liter cultures of HB-130 cells in LB medium, supplemented with ampicillin, were grown at 30°C as described by Wang et al, in J.Biol.Chem.2_60, 64 (1985). The expression of v-abl protein was induced by shifting the temperature to 42°C for 3-4 h. The bacterial cells were collected on ice, centrifuged and frozen in liquid nitrogen. The cell pellet was lysed as described by Ferguson et al. in J.Biol.Chem. 260, 3652 (1985) and in Biochem.J.
257, 321 (1989). Briefly, bacterial proteins were removed by differential ~solubilization with sodium deoxycholate, NaCl and ~-octylglucopyranoside..in different steps. Insoluble v-abl protein was solubilized with 2M FCSCN and dialyzed against 100 volumes of 50 mM Tris-HC1, pH 7.5, 1mM EDTA and 0.1 mM
- .L 1 -dithiothreitol (buffer A). The soluble proteins were separated by chromatography on a grepacked Mono-QTM anion exchange column in a f.p.l.c. system. The v-abl activity was eluted with a linear KC1 gradient (O-I M buffer A). Active fractions were pooled, made 50% in glycerol and stored in small aliquots at -20°C.
Myelin Basic Proteins phophor~lation assay The in vitro test with Myelin Basic Protein as substrate was carried out as follows; Protein phasphorylation was performed by incubating. ~0 ng of purified v-abl kinase, 1.5 pCi ~[~ 3~P]
ATP, 10 EzM cold ATP, 56 pM myelin basic protein in 50 ~1 of Tris-HC1 25 mM, pH 8.0, containing MgClz 10 mM, dithiothrei-tol 0.1 mM (kinase buffer) at 22°C.. The reaction was stopped by addition of equal ~rolumes of 2-fold concentrated Laem~nli electrophoresis buffer [U. K. Laem~nli, Nature (London) 230, 680 (1970). Samples were boiled again for 3 min and separated in SDS-PAGE (15% acrylamide). Gels were dried and exposed to autoradiographic films for 15-30 min at -70°C. Bands were located by autoradiography, excised from gel and counted in a liquid scintillation counter.
Autophosphorylation assay For the autophosphorylation assay the v-abl kinase was immunoprecipitated With antiphosphotyrosine antibodies and the resulting immunocomplex analyzed in 50 ~l of kinase buffer in the presence of 10 ~ZM ATP and 10 ~Ci [ ~-3~P]-ATP.
The reaction was stopped after 15 min at room temperature with boiling Laemmli buffer. Samples were boiled again for 3 min and separated in SDS-PAGE (8% acrylamide). Gels were dried and exposed to autoradiographic films for up to 3 h at -70°C. Bands were located by autoradiography, excised and counted as above.
W093/01182 z~~d:~o~s PCTlEP92/015f - i~ -In table I and II a representative compound of this invention is compared with the corresponding non hydroxylated analog which is encompassed by the general formula of patent ,.
application w~91/13055. The comparison shows that the introduction of an hydroxyl graup, while increasing only .
slightly the potency toward exogenous substrates significantly enhances the inhibitory activity on the autophosphorylation.
Table I. Myelin Pasic Protein phosphorylation assay.
ICso(~M) 5-hydroxy-3-j(3'-indolyl)methylen]-2-oxindole 0.4 3-[(3'-indolyl)methylen]-2-oxindole 0.6 Table II. Autophosphorylation assay.
ICSO(NM) 5-hydroxy-3-j(3'-indolyl)methylen]-2-oxindole 0.4 3-j(3'-indolyl)methylen]-2-oxindole : 10 In view of their high activity and low toxicity, the compounds of the invention can be used safely in medi-cine. For example, the approximate acute toxicity (LDP) of the compounds of the invention in the mouse, deter-mined by single administration of increasing doses and measured on the seventh day after the treatment was found to be negligible.
The compounds of the invention can be administered in a variety of dosage forms, e.g. orally, in the form of tablets, capsules, sugar or film-coated tablets, liquid solutions or suspensions; rectally, in the form of sup-positories; parenterally, e.g. intramuscularly, or by intravenous injection or infusion; or topically.
The dosage depends on the age, weight, conditions of the patient and administration route; for example, the dosage adopted for oral administration to adult humans may range from about 10 to about 150-200 mg per dose, from 1 to 5 times daily. Of course, these dosage regimens may be ad-justed to provide the optimal therapeutic response.
The invention includes pharmaceutical compositions com-prising a compound of formula (I) or a pharmaceutically acceptable salt thereof in association with a pharmaceu-tically acceptable excipient (which can be a carrier or diluent).
The invention also provides a pharmaceutical composition comprising a suitable carrier and/or diluent and, as an active principle, a compound selected from 3-[(1'-methyl-3'-indolyl)methylen]-2-oxindole, 3-[(3'-indolyl)methylen]-1-methyl-2-oxindole, and the pharmaceutically acceptable salts thereof, the compound or salt being either the Z- or E-diastereoisomer or a Z/E
mixture of diastereoisomers.
The compounds defined above as an active principle of the pharmaceutical composition are also provided by the invention for use as a tyrosine kinase inhibitor, as an anti-proliferative agent, as an anti-cancer agent or in the treatment of coronary a2tery disease.
The pharmaceutical compositions containing'the compounds of the invention are usually prepared following conven-tional methods and are administered in a pharmaceutical-ly suitable form.
For example, the solid oral forms may contain, together with the active compound, diluents, e.g., lactose, dex-trose, saccharose, cellulose, corn starch or potato starch; lubrican~.s, e.g. silica, talc, stearic acid, magnesium or calcium stearate, and/or polyethylene glyc-ols; binding agents, e.g. starches, arabic gums, gelatin, methylcellulose, carboxymethylcellulose or polyvinyl pyr-rolidone; disaggregating agents, e.g. a starch, alginic acid, alginates or sodium starch glycolate, effervescing mixtures; dyestuffs; sweeteners; wetting agents, such as ,v lecithin, polysorbates, laurylsulphates; and, in general, non-toxic and pharmacologically inactive substances used in pharmaceutical formulations. Said pharmaceutical pre-parations may be manufactured in known manner, for exacr~le, by means of mixing, granulating, tabletting, sugar-coat-ing or film-coating processes.
"'~ 93/01182 ~ ~ ~ ~ ~ ~ ~ PCTJEP'92/01569 The liquid dispersion for oral administration may be, e.g., syrups, emulsions and suspensions.
The syrup may contain as carrier, for example, sacchar-ose or saccharose with glycerine and/or mannitol and/or sorbitol.
The suspensions and the emulsions may contain as carrier, for example, a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl al-cohol.
The suspensions or solutions for intramuscular injections may contain, together with the active comnnmn~_ a r,h~.._ maceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oleate, glycols, e.g. propylene glyeol, and, if desired, a suitable amount of lidocaine hydro-chloride.
The solutions for intravenous injections or infusion may contain as carrier, for example; sterile water or, pre-ferably, they may be in the form of sterile, aqueous, isotonic saline solutions.
The suppositories may contain, together with the active compound, a pharmaceutically acceptable carrier, e.g.
cocoa-butter, polyethylene glycol, a palyoxyethylene sorbitan fatty acid ester surfactant or lecithin.
Compositions gor topical application, e.g. creaaas, lo-2S Lions, or pastes, can be prepared by admixing the active WO 93/01182 2 ~ ~ ~ ~ l~ ~ PCf/EP92/Ol~~'°' ingredient with a conventional oleaginous or emulsifying excipient.
A further object of the present invention is a combined method of treatment of cancer in mammals, including humans, in need of such treatment, said method compris-ing administering 1) a compound of formula (I), or a pharmaceutically ac-ceptable salt thereof, and 2) an additional antitumor agent, in amounts and close enough together in time sufficient to produce a the-rapeutically useful effect.
Object of the present invention is also to provide pro-ducts containing a compound of formula (I), or a pharma-ceutically acceptable salt, and an additional antitumor agent as a combined preparation for simultaneous, separ-ate or sequential use in anti-cancer therapy.
The term "antitumor agent" is meant to comprise both a single antltumor drug and °'cocktails" i.e. a mixture of such drugs, according to the clinical practice.
An titvanor agents that can be foranulated with a compound of the invention or, alternatively, can be administered in a combined method of treatment, are e.g. doxorubicin, daunomycin, eplrubicin, idarubicin, etoposide, flucro-uracil, mephalan, cyclophospha~aide, bleonycin, vinblas-~25 tin arid mitomycin or a mixture of two or snore thereof.
"'O 93/01182 1'~t'T/EP92/01569 7 _ T2-~e vompcundS of the invention can therefore be used in a treatment to ameliorate a cancer. They may be administered to~ a patient suffering from a cancer treatable with an antitumor agent, for example an anthracycline glycoside such as doxorubicin, daunomycin, epirubicin or idarubicin as mentioned above, together with the antiproliferative agent.
A compound of the invention and an antitumor agent such as an anthracycline glycoside can be administered to improve the ZO condition of a patient having a leukaemia such as myeloblastic leukaemia, lymphoma, sarcoma, neuroblastoma, wilm°s tumor or malignant neoplasm of the bladder, breast, lung or thyroid.
The following ekamples illustrate but do not limit the invention.
WO 93/01182 ~ ~ ~ ~ ~ Q8 - 1'GT/1rP92/Ol~'-'' Ex amp 1 a 1 5-Hydroxy-3-[(3'-indolyl)methylene]-2-oxindole ~I, ~R as defined, n = 0, R2 = R3 = R5 = R6 = H, R4 = 5-OH]
A solution of 3-indolecarboxaldehyde (145 mg, 1 mmol), 5-hydroxy-2-oxindole (149 mg, 1 mmol) and piperidine (60 mg, 0.7 mmol) in absolute ethanol (10 ml) is heated for 3 h at 60°C under nitrogen. Then the reaction mixture is chilled and evaporated under vacuum to dryness. The residue is submitted to column chromatography over silica gel using methylene-chloride/ethanol 4 % as eluant. Pure title compound is so ob-tained in 60 % yield (166 mg). - Alternatively, the reaction mixture is concentrated under vacuum and then chilled to 0-5°C, the precipitate filtered, the residue wash ed With ice-cooled ethanol and finally dried under vacuum. Compounds of higher purity are obtained by further crystallization from ethanol.
C17H12N202 requires: C 73.89 H 4.38 N 10.14 found: C 73.51 H 4.21 N 9.92 MS m/z: 276 IR cm 1 (7CBr): 3600-2500 (NH, 0H), 1650 (CO), 1600, 1580, 1530, 1480 (C=C) m.p. 293°C (dec.).
According to the above described procedure, the following compounds can be prepared:
5-carboxy-3-t(3'-indolyl)methylenel-2-oxindole;
5-amino-3-[3'-indolyl)methylene~-2-oxindole;
'"O 93/f1182 - 19 - PCT/EP92/O1S69 5-hydroxy-3-[(5'hydroxy-3'-indolyl)methylene]-2-oxindole;
NfS m/2 : ~ 282 IR cm-1 (KBr); 3600-2600 (NH, OH), 1655 (CO), 1605, 1585, 1535 (C=C) 5-hydroxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
3-L(5a7'-dihydroxy-3'-indolyl)methylene]-2-oxindole;
5-amino-3-C(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-amino-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-~(5'-carboxy-3'-indolyl)methylene]-2-oxindole;
5-amina-3-((7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-methoxy-3-((5'-tr~ethoxy-3'-indolyl)-methYlene'-2-oxindole;
5-acetoxy-3-~(5'-acetoxy-3'-indolyl)mettrylene,?-2-oxindole;
3-r(5'-carboxy-3'-indolyl)methylene]-2-oxindole, C18H12N203 requires: C 71.04; H 3.98; N 9.21 -found : C 70.83; H 4.6; N 8.85 MS m/z : 304;
m.p. > 330°dec.
IR cm 1 (KBr): 3600-3000 (NH), 3000-2100 (OH), 1710 (CO), 1640, 1620, 1600, 1550 Carom);
3-t(5'-amino-3°-indolyl)methylene~ -2-oxindole, C17H13N30 requires : C 74.14; H 4.71; N 15.26 ' found : C 73.88; H 4.51; N 14.91 2S MS m/z 275;
m.p. 250°dec.
IR cm 1 (KBr) . 3300, 2380 (NH), 1670 (CO), 1600, 1510 (C=C);
WO 93/o118z 2 ~ (~ ~ ~ ~ ~ Pc~r~EP9z,/ol~, '.
3-[(5'-nitro-3'-indolyl)methylen~ -2-oxindole, C17Hi1N303 requires : C 66.88; H 3.63; N 13.76 found : C 66.58; H 3.74; N 13.64 MS m/z 305;
m.p. > 350°C
IR cm-1 (KBr): 3350, 3230 (NH), 1680 (CO), 1620, 1605, 1580 (c=c), 1530, 1340 (N02);
3- ~(1'-methyl-3°-indolyl)methylen] -2-oxindole, C18H14N20 requires : C 78.81; H 5. I4; N 10.21 found : C 78.42; H 5.17; N 10.00 MS m/z 274 m.p. 230'C (dec.) IR cm 1 (KRr) . 3300--2000 (NH), 1680 (C0), 1610, 1600, 1570, 1500 (C=C);
3- [(3'-indolyl)methylen~-1-methyl-2-oxindole, C18H14N20 requires : C 78.81; N 5.14; N 10.21 found : C 78.61; H 5.16; N 10.23 MS m/z 274 m.p. 274°C
IR cm-1 (KBr) . 3220 (NH), 1675 (CO), 1605, 1500, 1490 (C=C).
93/01182 - 21 - . 2. ~ ~ ~ ~ ~ ~ P~.'T/EP92/01569 Example 2 5-Hydroxy-3-[(5'-hydroxy-3'-indolyl)methylene~-2-oxindole rI, R as defined, n = 1, R1 = R2 = R3 = R5 = R6 = H, R4=5-OH' The starting material for this de-etherification example is 5-methoxy-3- [(5'-methoxy-3'-indolyl)methylene~ -2-oxindole, which can be obtained according to the procedure described in Ex~ple 1.
To a stirred solution of 5-methoxy-3- C(5'-methoxy-3'-indolyl) methylene]-2-oxindole (310 mg, 1 mmol) in anhydrous dichloro-methane (10 ml) is added at -78°C under nitrogen, over a pe-riod of 10 min, a 1.0 M solution of boron tribromide in di-chloromethane (3 ml, 3 mmol). The resulting mixture is stir-red for another 1 h at -78°C and then allowed to warm to room temperature. After stirring for 1.5 h at 20-25°C. the mixture is cooled to -10°C and then quenched by the dropwise addition of water (10 ml) over a 10-min period. After addition of ethylacetate the organic layer is separated, washed with water, dried with Na2S04 and evaporated under vaeuum to dry-ness. The residue is crystallized from ethanol thus giving 198 mg of pure title compound (yield 70 %).
C17H12N203 requires: C 72.33 H 4.29 N 6.38 found: C 72.11 H 4.07 N 6.29 MS m/2: 282 IR cm 1 (KBr): 3600-2600 (NH, OH), 1655 (C0), 25' 1605, 1585, 1535 (C=C) , According to, the above descx°ibed procedure and starting from the corresponding methyletlaer, the. hydroxyl compounds men-tioned in Example 1 can be obtained.v wc~ 9momsz '~,' ~ ~ ~. ~ ~ $ ~~crm~ziol~ ._:_, Example 3 5-Acetoxy-3- ~(5'-acetoxy-3'-indolyl)methylene~-2-oxindole ~I, R as defined, n = 1, R1 = Ac, R4 = 5-OAc, R2=R3=R5=R6=H~
The starting material for this acylation example is 5-hydroxy-3-[(5'-hydroxy-3'-indolyl)methylene~ -2-oxindole, which may be obtained according to the procedure described in Examples 1 an d 2 .
To a cooled solution of 5-hydroxy-3-[(5'-hydroxy-3'-indolyl) methylene'-2-oxindole (282 mg, 1 mmol) in dry pyridine (0.5 ml) is added acetic anhydride (306 mg, 3 mmol) and the.mix-ture maintained at 0-5°C overnight. Thereupon the mixture concentrated under vacuum, the residue dissolved in dichloro-methane, the organic layer washed with water and then evapor-ated under reduced pressure. The crude product is crystallized from chlaroform/methanol to yield pure title compound in 80%
yield (301 mg).
C21H16N205 requires: C 67:02 H 4.29 N 7.44 found: C 66.91 H 4.05 N 7.29 MS m/z: 376 IR cm 1 (KRr): 3600-3200 (NH), 1750 (CH3C00), 1650 (CONH), 1600, 1580, 1530 (C=C) According to the above described procedure, the hydroxyl com-pounds obtained in~Example l and 2 can be transformed into the eorrespondfng.~~CZ-C6 alkanoyloxyd~rivativese .
7 93/01182 '~ ~ ~ ~ fl ~ ~ PC.'1'1EP92/01569 Example 4 5-Nitro-3-indolealdehyde ~I, n -_ 0, R2 = 5-N02, R3 = H;
A mixture of N-methylformanilide~(176 mg, 1.3 mmol) and phos-S phorous oxychloride (199 mg, 1.3 mmol) is stirred for 15 min at 20-25°C under nitrogen. then a solution of 5-nitroindole (162 mg, 1 mmol) in 1,2-dichloroethane (S ml) is added and the mixture heated to reflux for 3 h. After cooling the mix-ture is poured onto iced water, the precipitate filtered off 1p and washed with water. Thereupon the residue is chromato-graphed over silica gel using benzene/ethyl acetate as eluant.
Thus pure title compound is obtained in 80 % yield (152 mg).
CgH6N203 requires: C 56.85 H 3.18 N 14.73 found: C 56.79 H 3.01 N 14.51 15 MS m/z: 190 IR cm-1 (KHr): 3140, 3090 (NH), 1650 (CO), 1511, 1345 (N02) These vitro-intermediates besides leading to final products of formula (I) with R 2 = N02 give also rise to final proudcts 20 with R 2 = NH2 obtainable from the former by reduction.
~y proceding analogously, the following protected interme-diates can be prepared, which after deprotection at a suit-able stage of the synthesis give also rise to final product of formula (I) with free carboxyl (R 2 = COON) and free hydr-25 oxyl (R 2 = OH) respectively:
3-carbcx~ethoxy-3-indolealdehyde; and 3-methoxy-3-indolealdehyde.
WO 93J01182 ~ ~ ~ ~ ~~ ~ ~ c..., PCE'/EP92/Oi., _ ~4 _ Example 5 Tablets each weighing 0.150 g and containing 25 mg of the active substance, can be manufactured as follows:
composition (for 10,000 tablets):
5-Hydroxy-3-[(3'-indolyl)methylene3-2-oxindole 250 g Lactose 800 g Corn starch 415 g Talc powder 30 g P:agnesium stearate 5 g Th a 5-hydroxy-3-~(3'-indolyl}methylene~-2-oxindole, the-lactose and half the corn starch are mixed; the mixture is then forc-ed through a sieve of 0.5 mm mesh size.
Corn starch (10 g} is suspended in warm water (90 ml) and the resulting paste is used to granulate the powder. The granul-ate is dried, comminuted on a sieve of 1.4 mm mesh size, then the remaining quantity of starch, talc and magnesium stearate are added, carefully mixed and processed into tablets.
~='d!O 93/0i182 ~fT/EP92/Ole' ~~
Example 6 Capsules, each dosed at 0.200 g and containing 20 mg of the active substance can be prepared.
Composition for 500 capsules:
3-[(5'-amino-3°-indolyl)methylene~ -2-oxindole 10 g Lactose 80 g Corn starch 5 g , Magnesium stearate 5 g This formulation is encapsulated in two-piece hard gelatin capsules and dosed at 0.200 g for each capsule.
Claims (11)
1. A compound of formula (I) in which R4 is hydrogen, hydroxy, C1-C5 alkoxy, C2-C6 alkanoyloxy, carboxy, nitro or NHR7, wherein R7 is hydrogen or C1-C6, alkyl;
R5, is hydrogen, C1-C6 alkyl er halogen; and R6 is hydrogen or C1-C6 alkyl;
n is zero, 1 or 2;
R1 is hydrogen, C1-C6 alkyl or C2-C6 alkanoyl;
R2 is hydrogen, C1-C6 alkyl, halogen, cyano, carboxyl, nitro or -NHR7 in which R7 is as defined above;
R3 is hydrogen, C1-C6, alkyl ar C2-C6 alkanoyl; or a pharmaceutically acceptable salt thereof; and wherein, (i) when, at the same time, R2 is hydrogen, C1-C6 alkyl, halogen or cyano and R3 is hydrogen, R1 and n being as defined above, then at least one of R4, R5 and R6 is other than hydrogen; and (ii) when n is zero and R3 and R6 are each hydrogen or CH3, then at least one of R2, R4 and R5 is other than hydrogen.
R5, is hydrogen, C1-C6 alkyl er halogen; and R6 is hydrogen or C1-C6 alkyl;
n is zero, 1 or 2;
R1 is hydrogen, C1-C6 alkyl or C2-C6 alkanoyl;
R2 is hydrogen, C1-C6 alkyl, halogen, cyano, carboxyl, nitro or -NHR7 in which R7 is as defined above;
R3 is hydrogen, C1-C6, alkyl ar C2-C6 alkanoyl; or a pharmaceutically acceptable salt thereof; and wherein, (i) when, at the same time, R2 is hydrogen, C1-C6 alkyl, halogen or cyano and R3 is hydrogen, R1 and n being as defined above, then at least one of R4, R5 and R6 is other than hydrogen; and (ii) when n is zero and R3 and R6 are each hydrogen or CH3, then at least one of R2, R4 and R5 is other than hydrogen.
2. A compound of formula (I), according to claim 1, wherein R4 is hydroxy, amino, nitro or carboxy and R5 and R6 are hydrogen;
R1 is hydrogen or C1-C6 alkyl;
n is zero or 1;
R2 is hydrogen, carboxy, amino or nitro;
R3 is hydrogen; and the pharmaceutically acceptable salts thereof.
R1 is hydrogen or C1-C6 alkyl;
n is zero or 1;
R2 is hydrogen, carboxy, amino or nitro;
R3 is hydrogen; and the pharmaceutically acceptable salts thereof.
3. A compound of formula (I), according to claim 1, wherein R4 is hydroxy, amino or carboxy; and R5 and R6 are hydrogen; n is 0 or 1; R1 is hydrogen; R2 is hydrogen, amino or carboxy; R3 is hydrogen; and the pharmaceutically acceptable salts thereof.
4. A compound selected from the group consisting of the following which may be either a Z- or E-diastereoisomer or a Z,E-mixture of said diastereoisomers:
5-hydroxy-3-[(3'-indolyl)methylene]-2-oxindole;
3-[(5'-carboxy-3'-indolyl)methylene]-2-oxindole;
3-[(5'-amino-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(3'-indolyl)methylene]-2-oxindole;
5-amino-3-[(3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-amino-3-[(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-amino-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(5'-hydroxy-:3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-carboxy-3'-indolyl)methylene]-2-oxindole ;
5-amino-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-methoxy-3-[(5'-methoxy-3'-indolyl)-methylene]-2-oxindole;
5-acetoxy-3-[(5'-acetoxy-3'-indolyl)methylene]-2-oxindole;
3-[(5'-nitro-3'-indolyl)methylen]-2-oxindole;
and; the pharmaceutically acceptable salts thereof.
5-hydroxy-3-[(3'-indolyl)methylene]-2-oxindole;
3-[(5'-carboxy-3'-indolyl)methylene]-2-oxindole;
3-[(5'-amino-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(3'-indolyl)methylene]-2-oxindole;
5-amino-3-[(3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-amino-3-[(5'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-amino-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(5'-hydroxy-:3'-indolyl)methylene]-2-oxindole;
5-hydroxy-3-[(5'-carboxy-3'-indolyl)methylene]-2-oxindole ;
5-amino-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-carboxy-3-[(7'-hydroxy-3'-indolyl)methylene]-2-oxindole;
5-methoxy-3-[(5'-methoxy-3'-indolyl)-methylene]-2-oxindole;
5-acetoxy-3-[(5'-acetoxy-3'-indolyl)methylene]-2-oxindole;
3-[(5'-nitro-3'-indolyl)methylen]-2-oxindole;
and; the pharmaceutically acceptable salts thereof.
5. A process for the preparation of a compound of formula (I), or a pharmaceutically acceptable salt thereof, according to claim 1 the process comprising condensation of an aldehyde of formula (II) wherein R1, R2, R3 and n are as defined in claim 1, with a compound of formula (III) wherein R4, R5 and R6 are as defined in claim 1 and, if desired, converting a compound of formula (I) into another compound of formula (I), and/or, if desired, converting a compound of formula (I) into a pharmaceutically acceptable salt thereof, and/or, if desired, converting a salt into a free compound, and/or, if desired, separating a mixture of isomers of a compound of formula (I) into the single isomers.
6. A pharmaceutical composition comprising a compound or salt according to any one of claims 1 to 4 and a pharmaceutically acceptable carrier or diluent.
7. A pharmaceutical composition comprising a compound selected from 3-[(1'-methyl-3'-indelyl)methylen] -2-oxindole, 3[(3'-indolyl)methylen]-1-methyl-2-oxindole and the pharmaceutically acceptable salts thereof; the compound or salt being a Z- or E-diastereoisomer or a Z/E mixture of diastereoisomers, and a pharmaceutically acceptable carrier or diluent.
8. A compound or salt according to any one of claims 1 to 4 or a compound or salt as defined in claim 7, for use as a tyrosine kinase inhibitor.
9. A compound or salt according to any one of claims 1 to 4 or a compound or salt, as defined in claim 7, for use as an anti-proliferative agent.
10. A compound or salt according to any one of claims 1 to 4 or a compound or salt as defined in claim 7, for use as an anti-cancer agent or in the treatment of coronary artery disease.
11. A combined preparation comprising a compound or salt according to any one of claims 1 to 4 or a compound or salt as defined in claim 7 and an antitumor agent in a dosage format adapted for simultaneous, separate or sequential administration in anti-cancer therapy.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB919115160A GB9115160D0 (en) | 1991-07-12 | 1991-07-12 | Methylen-oxindole derivatives and process for their preparation |
| GB9115160.5 | 1991-07-12 | ||
| PCT/EP1992/001569 WO1993001182A1 (en) | 1991-07-12 | 1992-07-10 | Methylen-oxindole derivatives and process for their preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2091058A1 CA2091058A1 (en) | 1993-01-13 |
| CA2091058C true CA2091058C (en) | 2004-03-30 |
Family
ID=10698323
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002091058A Expired - Fee Related CA2091058C (en) | 1991-07-12 | 1992-07-10 | Methylen-oxindole derivatives and process for their preparation |
Country Status (22)
| Country | Link |
|---|---|
| US (1) | US5409949A (en) |
| EP (2) | EP0552329B1 (en) |
| JP (1) | JP3188701B2 (en) |
| KR (1) | KR930702337A (en) |
| AT (1) | ATE206420T1 (en) |
| AU (1) | AU656015B2 (en) |
| CA (1) | CA2091058C (en) |
| DE (1) | DE69232093T2 (en) |
| DK (1) | DK0552329T3 (en) |
| ES (1) | ES2165357T3 (en) |
| FI (1) | FI931061A0 (en) |
| GB (1) | GB9115160D0 (en) |
| HU (1) | HUT67496A (en) |
| IE (1) | IE922253A1 (en) |
| IL (1) | IL102383A (en) |
| MX (1) | MX9204008A (en) |
| NZ (1) | NZ243454A (en) |
| PT (1) | PT552329E (en) |
| RU (1) | RU2072989C1 (en) |
| TW (1) | TW202444B (en) |
| WO (1) | WO1993001182A1 (en) |
| ZA (1) | ZA925169B (en) |
Families Citing this family (49)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9004483D0 (en) * | 1990-02-28 | 1990-04-25 | Erba Carlo Spa | New aryl-and heteroarylethenylene derivatives and process for their preparation |
| US5587385A (en) * | 1991-12-24 | 1996-12-24 | Farmitalia Carlo Erba S.R.L. | Arylidene-heterocyclic derivatives and process for their preparation |
| GB9313638D0 (en) * | 1993-07-01 | 1993-08-18 | Erba Carlo Spa | Arylidene and heteroarylidene oxindole derivatives and process for their preparation |
| GB9326136D0 (en) * | 1993-12-22 | 1994-02-23 | Erba Carlo Spa | Biologically active 3-substituted oxindole derivatives useful as anti-angiogenic agents |
| GB9406137D0 (en) * | 1994-03-28 | 1994-05-18 | Erba Carlo Spa | N-substituted beta-aryl- and betaheteroaryl-alpha-cyanoacrylamide derivatives and process for their preparation |
| GB9412719D0 (en) * | 1994-06-24 | 1994-08-17 | Erba Carlo Spa | Substituted azaindolylidene compounds and process for their preparation |
| GB9501567D0 (en) * | 1995-01-26 | 1995-03-15 | Pharmacia Spa | Hydrosoluble 3-arylidene-2-oxindole derivatives as tyrosine kinase inhibitors |
| GB9507298D0 (en) * | 1995-04-07 | 1995-05-31 | Pharmacia Spa | Substituted indolylmethylene-oxindale analogues as tyrosine kinase inhibitors |
| US5880141A (en) * | 1995-06-07 | 1999-03-09 | Sugen, Inc. | Benzylidene-Z-indoline compounds for the treatment of disease |
| US6147106A (en) | 1997-08-20 | 2000-11-14 | Sugen, Inc. | Indolinone combinatorial libraries and related products and methods for the treatment of disease |
| US6906093B2 (en) | 1995-06-07 | 2005-06-14 | Sugen, Inc. | Indolinone combinatorial libraries and related products and methods for the treatment of disease |
| GB9610964D0 (en) * | 1996-05-24 | 1996-07-31 | Pharmacia & Upjohn Spa | Substituted tetralylmethylen-oxindole analogues as tyrosine kinase inhibitors |
| AU4155697A (en) * | 1996-08-23 | 1998-03-06 | Sugen, Inc. | Indolinone combinatorial libraries and related products and methods for the treatment of disease |
| AU7622698A (en) * | 1996-12-05 | 1998-06-29 | Sugen, Inc. | Use of indolinone compounds as modulators of protein kinases |
| US6051597A (en) * | 1997-06-13 | 2000-04-18 | Merck & Co., Inc. | Indolylquinones as antidiabetic agents |
| US20040067531A1 (en) * | 1997-08-20 | 2004-04-08 | Sugen, Inc. | Methods of modulating protein tyrosine kinase function with substituted indolinone compounds |
| GB9718913D0 (en) | 1997-09-05 | 1997-11-12 | Glaxo Group Ltd | Substituted oxindole derivatives |
| US6531502B1 (en) | 1998-01-21 | 2003-03-11 | Sugen, Inc. | 3-Methylidenyl-2-indolinone modulators of protein kinase |
| AU760046B2 (en) | 1998-02-27 | 2003-05-08 | United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services, The | Disubstituted lavendustin a analogs and pharmaceutical compositions comprising the analogs |
| US6102942A (en) | 1998-03-30 | 2000-08-15 | Endovascular Technologies, Inc. | Stent/graft deployment catheter with a stent/graft attachment mechanism |
| US6569868B2 (en) | 1998-04-16 | 2003-05-27 | Sugen, Inc. | 2-indolinone derivatives as modulators of protein kinase activity |
| AU5468499A (en) * | 1998-08-04 | 2000-02-28 | Sugen, Inc. | 3-methylidenyl-2-indolinone modulators of protein kinase |
| US6680335B2 (en) | 1998-09-28 | 2004-01-20 | Sugen, Inc. | Methods of modulating protein tyrosine kinase function with substituted indolinone compounds |
| AU760039B2 (en) | 1998-12-17 | 2003-05-08 | F. Hoffmann-La Roche Ag | 4-aryloxindoles as inhibitors of JNK protein kinases |
| BR9916324A (en) | 1998-12-17 | 2001-10-02 | Hoffmann La Roche | 4,5-pyrazinoxindoles as protein kinase inhibitors |
| US6153634A (en) * | 1998-12-17 | 2000-11-28 | Hoffmann-La Roche Inc. | 4,5-azolo-oxindoles |
| KR100649925B1 (en) | 1998-12-17 | 2007-02-28 | 에프. 호프만-라 로슈 아게 | 4-alkenyl and alkynyl oxindoles as inhibitors of cyclin-dependent kinases, in particular cdk2 |
| JP2000247949A (en) * | 1999-02-26 | 2000-09-12 | Eisai Co Ltd | Indole compound containing sulfonamide |
| GB9904933D0 (en) | 1999-03-04 | 1999-04-28 | Glaxo Group Ltd | Compounds |
| US6492398B1 (en) | 1999-03-04 | 2002-12-10 | Smithkline Beechman Corporation | Thiazoloindolinone compounds |
| US6624171B1 (en) | 1999-03-04 | 2003-09-23 | Smithkline Beecham Corporation | Substituted aza-oxindole derivatives |
| CA2368041A1 (en) * | 1999-03-24 | 2000-09-28 | Sugen, Inc. | Indolinone compounds as kinase inhibitors |
| US6689806B1 (en) | 1999-03-24 | 2004-02-10 | Sugen, Inc. | Indolinone compounds as kinase inhibitors |
| US6878733B1 (en) | 1999-11-24 | 2005-04-12 | Sugen, Inc. | Formulations for pharmaceutical agents ionizable as free acids or free bases |
| US6313310B1 (en) | 1999-12-15 | 2001-11-06 | Hoffmann-La Roche Inc. | 4-and 5-alkynyloxindoles and 4-and 5-alkenyloxindoles |
| CA2399001C (en) | 2000-02-03 | 2010-07-20 | Eisai Co., Ltd. | Integrin expression inhibitor |
| ES2290117T3 (en) * | 2000-02-15 | 2008-02-16 | Sugen, Inc. | PROTEIN QUINASE 2-INDOLIN INHIBITORS REPLACED WITH PIRROL. |
| US6620818B1 (en) | 2000-03-01 | 2003-09-16 | Smithkline Beecham Corporation | Method for reducing the severity of side effects of chemotherapy and/or radiation therapy |
| MY128450A (en) | 2000-05-24 | 2007-02-28 | Upjohn Co | 1-(pyrrolidin-1-ylmethyl)-3-(pyrrol-2-ylmethylidene)-2-indolinone derivatives |
| WO2001094312A2 (en) * | 2000-06-02 | 2001-12-13 | Sugen, Inc. | Indolinone derivatives as protein kinase/phosphatase inhibitors |
| AR042586A1 (en) | 2001-02-15 | 2005-06-29 | Sugen Inc | 3- (4-AMIDOPIRROL-2-ILMETILIDEN) -2-INDOLINONE AS INHIBITORS OF PROTEIN KINASE; YOUR PHARMACEUTICAL COMPOSITIONS; A METHOD FOR THE MODULATION OF THE CATALYTIC ACTIVITY OF PROTEINQUINASE; A METHOD TO TREAT OR PREVENT AN AFFECTION RELATED TO PROTEINQUINASE |
| US6599902B2 (en) | 2001-05-30 | 2003-07-29 | Sugen, Inc. | 5-aralkysufonyl-3-(pyrrol-2-ylmethylidene)-2-indolinone derivatives as kinase inhibitors |
| AR038957A1 (en) * | 2001-08-15 | 2005-02-02 | Pharmacia Corp | COMBINATION THERAPY FOR CANCER TREATMENT |
| BR0213185A (en) | 2001-10-10 | 2004-09-14 | Sugen Inc | 3- [4- (Substituted Heterocyclyl) -pyrrol-2-ylmethylidene] 2-indolinone derivatives as kinase inhibitors |
| US20050119303A1 (en) * | 2002-03-05 | 2005-06-02 | Eisai Co., Ltd | Antitumor agent comprising combination of sulfonamide-containing heterocyclic compound with an angiogenesis inhibitor |
| US20050107819A1 (en) * | 2003-11-14 | 2005-05-19 | Medtronic Vascular, Inc. | Catheter with a sectional guidewire shaft |
| US8772269B2 (en) * | 2004-09-13 | 2014-07-08 | Eisai R&D Management Co., Ltd. | Use of sulfonamide-including compounds in combination with angiogenesis inhibitors |
| WO2010149943A1 (en) * | 2009-06-25 | 2010-12-29 | Oncorel Ab | New compounds and medical use |
| CN103274988B (en) * | 2013-06-20 | 2016-04-13 | 河北大学 | 3-(cyano group-p-nitrobenzoyl support group)-2-indolone derivatives and preparation method and use thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3310891A1 (en) * | 1983-03-25 | 1984-09-27 | Boehringer Mannheim Gmbh, 6800 Mannheim | NEW INDOLINON (2) DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF, MEDICINAL PRODUCTS CONTAINING THESE COMPOUNDS AND INTERMEDIATE PRODUCTS |
| JPH03213847A (en) * | 1990-01-18 | 1991-09-19 | Konica Corp | Silver halide photographic sensitive material |
| GB9004483D0 (en) * | 1990-02-28 | 1990-04-25 | Erba Carlo Spa | New aryl-and heteroarylethenylene derivatives and process for their preparation |
-
1991
- 1991-07-12 GB GB919115160A patent/GB9115160D0/en active Pending
-
1992
- 1992-07-02 IL IL102383A patent/IL102383A/en not_active IP Right Cessation
- 1992-07-06 TW TW081105346A patent/TW202444B/zh active
- 1992-07-06 NZ NZ243454A patent/NZ243454A/en unknown
- 1992-07-08 MX MX9204008A patent/MX9204008A/en not_active IP Right Cessation
- 1992-07-10 DE DE69232093T patent/DE69232093T2/en not_active Expired - Fee Related
- 1992-07-10 JP JP50198193A patent/JP3188701B2/en not_active Expired - Fee Related
- 1992-07-10 AU AU22777/92A patent/AU656015B2/en not_active Ceased
- 1992-07-10 DK DK92914619T patent/DK0552329T3/en active
- 1992-07-10 US US07/987,280 patent/US5409949A/en not_active Expired - Lifetime
- 1992-07-10 HU HU9300723A patent/HUT67496A/en unknown
- 1992-07-10 EP EP92914619A patent/EP0552329B1/en not_active Expired - Lifetime
- 1992-07-10 ES ES92914619T patent/ES2165357T3/en not_active Expired - Lifetime
- 1992-07-10 EP EP19920111757 patent/EP0525472A3/en active Pending
- 1992-07-10 AT AT92914619T patent/ATE206420T1/en not_active IP Right Cessation
- 1992-07-10 RU RU9293004893A patent/RU2072989C1/en active
- 1992-07-10 WO PCT/EP1992/001569 patent/WO1993001182A1/en active IP Right Grant
- 1992-07-10 PT PT92914619T patent/PT552329E/en unknown
- 1992-07-10 IE IE225392A patent/IE922253A1/en not_active Application Discontinuation
- 1992-07-10 ZA ZA925169A patent/ZA925169B/en unknown
- 1992-07-10 CA CA002091058A patent/CA2091058C/en not_active Expired - Fee Related
-
1993
- 1993-03-10 FI FI931061A patent/FI931061A0/en not_active Application Discontinuation
- 1993-03-12 KR KR1019930700756A patent/KR930702337A/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| EP0525472A3 (en) | 1993-02-24 |
| IL102383A (en) | 1997-09-30 |
| HUT67496A (en) | 1995-04-28 |
| GB9115160D0 (en) | 1991-08-28 |
| EP0552329A1 (en) | 1993-07-28 |
| JPH06501494A (en) | 1994-02-17 |
| ES2165357T3 (en) | 2002-03-16 |
| JP3188701B2 (en) | 2001-07-16 |
| KR930702337A (en) | 1993-09-08 |
| AU2277792A (en) | 1993-02-11 |
| PT552329E (en) | 2002-02-28 |
| US5409949A (en) | 1995-04-25 |
| DE69232093D1 (en) | 2001-11-08 |
| MX9204008A (en) | 1993-01-01 |
| WO1993001182A1 (en) | 1993-01-21 |
| DK0552329T3 (en) | 2001-11-12 |
| NZ243454A (en) | 1995-02-24 |
| DE69232093T2 (en) | 2002-05-16 |
| IL102383A0 (en) | 1993-01-14 |
| FI931061A (en) | 1993-03-10 |
| EP0525472A2 (en) | 1993-02-03 |
| EP0552329B1 (en) | 2001-10-04 |
| TW202444B (en) | 1993-03-21 |
| AU656015B2 (en) | 1995-01-19 |
| RU2072989C1 (en) | 1997-02-10 |
| IE922253A1 (en) | 1993-01-13 |
| HU9300723D0 (en) | 1993-06-28 |
| ZA925169B (en) | 1993-04-28 |
| ATE206420T1 (en) | 2001-10-15 |
| CA2091058A1 (en) | 1993-01-13 |
| FI931061A0 (en) | 1993-03-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2091058C (en) | Methylen-oxindole derivatives and process for their preparation | |
| US5397787A (en) | Vinylene-azaindole derivatives | |
| EP0470221B1 (en) | New arylethenylene derivatives and process for their preparation | |
| EP0987263B1 (en) | Arylidene and heteroarylidene oxindole derivatives as tyrosine kinase inhibitors | |
| EP0700388B1 (en) | Substituted beta-aryl and beta-heteroaryl-alpha-cyanoacrylamide derivatives as tyrosine kinase inhibitors | |
| EP0558950B1 (en) | 3-aryl-glycidic ester derivatives and process for their preparation | |
| US5639884A (en) | Arylidene-heterocyclic derivatives and process for their preparation | |
| US5587385A (en) | Arylidene-heterocyclic derivatives and process for their preparation | |
| CA2163115C (en) | N-substituted beta-aryl- and beta-heteroaryl-alpha-cyanoacrylamide derivatives and process for their preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |