JP2007523850A - Indole derivatives with apoptosis-inducing action - Google Patents

Abstract

  The present invention relates to an indole derivative used as a medicament for the treatment of tumor diseases, particularly in the case of drug resistance to other active ingredients and in the case of metastatic carcinoma.

Description

  The invention is used as a medicament for the treatment of tumor diseases with improved biological action, compatibility and oral bioavailability, in particular in drug resistance to another agent and in metastatic carcinoma. The present invention relates to a novel indole derivative.

  Treatment of cancer diseases has great significance in medicine. There is a global need for cancer therapies that are patient-friendly and that work well for targeted therapy. This is reflected in the many chemical studies that have recently appeared in the field of applied oncology or basic research for cancer therapy.

  The action of tumor inhibitors is based on various mechanisms in this regard and is only partially known. It is not unusual to find a new mechanism of action for known tumor drugs. This is also expected for the compounds according to the invention. Many tumor drugs block the cell division mechanism of cells, block the supply of nutrients and oxygen to the tumor (anti-angiogenesis), inhibit metastasis, suppress the reception and transfer of growth signals in tumor cells or programmed cells Acts through mechanisms such as the elimination of tumor cells in death (apoptosis).

  Based on various mechanisms of action, especially interaction with various intracellular targets, clinically relevant cytostatic drugs are administered in combination to obtain a synergistic therapeutic effect.

  Indole derivatives are widely used as pharmacokinetically effective compounds and as building blocks for synthesis in medicinal chemistry.

  In the documents WO 99/51224 A1 and WO 01/22954 A1, in a number of groups, in particular 2-, 3-, 4- and 8-quinoline groups or 2-, 3-, 4-, 5- and 6-pyridine groups. Indol-3-yl derivatives having an antitumor action which may be substituted are described. In Example 60, a 2-methyl-8-quinolinyl group is mentioned as a substituent on the amide group. However, no biological properties are listed.

  WO 99/55696 A1 describes substituted hydroxyindoles as phosphodiesterase 4 inhibitors. The antitumor activity for the compounds according to the invention is neither described nor suggested.

  WO 02/08225 A1 describes 2- (1H-indol-3-yl) -2-oxoacetamide derivatives that have antitumor activity against solid tumors. However, specific examples having a quinoline group, a pyridopyrazine group or an indazolyl group are not mentioned.

  Patent document WO00 / 67802 describes indole-3-glyoxylamide substituted with a long chain fatty acid as a potential antitumor agent. However, specific examples having a quinoline group, a pyridopyrazine group or an indazolyl group are not mentioned. Also, no biological data is shown in the examples.

  W. -T. Li et al. (J. Med. Chem. 2003, 46, pages 1706 et seq.) Describe N-heterocyclic indolylglyoxylamide as an active compound for oral use having antitumor activity. However, the mechanism of action is not described.

  Patent application WO 03/022280 describes 3-glyoxylamidoindole and its use as a medicament for antitumor treatment. The general formula includes, among others, 6-quinoline derivatives. In addition, two examples with 6-quinoline groups are given as examples, which are shown in the biological results. However, specific examples having a pyridopyrazine group or an indazolyl group are not given.

  In the US application US 03/0181482 A1, novel indolylglyoxylamides are described. The compounds according to the invention are described in this connection as antitumor agents and angiogenesis inhibitors having cytotoxic activity. In addition, 6-quinoline derivatives are shown as examples (compound 3; page 10) and anti-proliferative data (see Tables 1a, 1b on page 19) and anti-angiogenic properties (see page 20) ). However, specific examples having a pyridopyrazine group or an indazolyl group are not given.

  Applicant's WO 02/10152 A2 already describes another class of indole derivatives for the treatment of tumors. In particular, there is the agent N- (2-methyl-6-quinolyl)-[1- (4-chlorobenzyl) -indol-3-yl] -glyoxylic acid amide, its anti-proliferation in various tumor cell lines. Tested for action.

  Tested compounds that bind to microtubules (paclitaxel, vincristine) or inhibit topoisomerase II (doxorubicin, etoposide, mitoxantrone) are currently in cancer therapy, especially cancer therapy for breast cancer, ovarian cancer, stomach cancer, lung cancer In Kaposi's sarcoma and in leukemia. However, its use is limited by serious neurological, gastrointestinal, cardiovascular and liver side effects, even with the development of drug resistance.

  The problem underlying the present invention is now to provide a cytotoxic agent with a combined mechanism of action suitable for treating many tumors, particularly in agent resistance to another drug and in metastatic carcinoma. That is.

  The problem is solved by the general formula I

［式中、
Ｒは、Ｎ、Ｏ及びＳの群から選択される１つ以上のヘテロ原子を有する飽和、不飽和又は芳香族の、アミド窒素と直接結合された、置換又は非置換のＣ_２〜Ｃ_１４−複素環を表し、その際、該複素環は有利には
（ｉ）非置換又は置換の５−、６−、７−キノリル、
（ｉｉ）非置換又は置換の２−、３−、６−、７−及び８−ピリドピラジニル、
（ｉｉｉ）非置換又は置換の３−、４−、５−、６−及び７−インダゾリル、
（ｉｖ）非置換又は置換の２−、３−、４−、５−及び６−ピリジル、
（ｖ）非置換又は置換の３−、４−及び５−イソキサゾリル、
（ｖｉ）非置換又は置換の３−、４−及び５−イソチアゾリル
を意味し、
Ｒ１は非置換又は置換のアルキル−アリールを意味し、
Ｒ２は
（ｉ）水素、
（ｉｉ）非置換又は置換のＣ_１〜Ｃ_６−アルキル
を意味し、
Ｒ３〜Ｒ６は
（ｉ）水素、
（ｉｉ）非置換又は置換のＣ_１〜Ｃ_６−アルキル、
（ｉｉｉ）非置換又は置換のＣ_３〜Ｃ_７−シクロアルキル、
（ｉｖ）アミノ、モノ−Ｃ_１〜Ｃ_４−アルキルアミノ、ジ−Ｃ_１〜Ｃ_４−アルキルアミノ、
（ｖ）ハロゲン、
（ｖｉ）１つ以上のフッ素原子で置換されたＣ_１〜Ｃ_４−アルキル、有利にはトリフルオロメチル基、
（ｖｉｉ）シアノ、直鎖状又は分枝鎖状のシアノ−Ｃ_１〜Ｃ_６−アルキル、
（ｖｉｉｉ）Ｃ_１〜Ｃ_６−アルキルカルボニル、
（ｉｘ）カルボキシル、Ｃ_１〜Ｃ_４−アルコキシカルボニル、カルボキシ−Ｃ_１〜Ｃ_６−アルキル又はＣ_１〜Ｃ_６−アルコキシカルボニル−Ｃ_１〜Ｃ_６−アルキル、
（ｘ）ヒドロキシ、
（ｘｉ）Ｃ_１〜Ｃ_６−アルコキシ、
（ｘｉｉ）アリール−Ｃ_１〜Ｃ_４−アルコキシ、有利にはベンジルオキシ、
（ｘｉｉｉ）Ｃ_１〜Ｃ_６−アルコキシカルボニルアミノ、Ｃ_１〜Ｃ_６−アルコキシカルボニルアミノ−Ｃ_１〜Ｃ_６−アルキル
を意味し、
Ｒ７はＣ_１〜Ｃ_６−アルキルカルボニル、有利にはアセチル、プロピオニル、Ｃ_１〜Ｃ_６−アルコキシカルボニル、有利にはメトキシカルボニル、エトキシカルボニル、プロポキシカルボニルを意味し、かつ
Ｘ、Ｙは酸素又は硫黄を意味する］のインドール誘導体、それらの互変異性体、立体異性体、例えばジアステレオマー及びエナンチオマー並びにそれらの生理学的に認容性の塩によって解決される。

[Where:
R is, N, saturated with one or more heteroatoms selected from the group of O and S, unsaturated or aromatic, coupled directly to the amide nitrogen, substituted or unsubstituted C ₂ -C ₁₄ - Represents a heterocycle, in which the heterocycle is preferably (i) unsubstituted or substituted 5-, 6-, 7-quinolyl,
(Ii) unsubstituted or substituted 2-, 3-, 6-, 7- and 8-pyridopyrazinyl,
(Iii) unsubstituted or substituted 3-, 4-, 5-, 6- and 7-indazolyl,
(Iv) unsubstituted or substituted 2-, 3-, 4-, 5- and 6-pyridyl,
(V) unsubstituted or substituted 3-, 4- and 5-isoxazolyl,
(Vi) means unsubstituted or substituted 3-, 4- and 5-isothiazolyl,
R1 means unsubstituted or substituted alkyl-aryl,
R2 is (i) hydrogen,
(Ii) unsubstituted or substituted _C 1 -C ₆ - refers to an alkyl,
R3 to R6 are (i) hydrogen,
(Ii) unsubstituted or substituted _C 1 -C ₆ - alkyl,
(Iii) unsubstituted or substituted _C 3 -C ₇ - cycloalkyl,
(Iv) amino, _mono--C 1 -C ₄ - alkylamino, di _-C 1 -C ₄ - alkylamino,
(V) halogen,
(Vi) C ₁ -C ₄ -alkyl, preferably a trifluoromethyl group, substituted with one or more fluorine atoms,
(Vii) cyano, straight-chain or branched _cyano--C 1 -C ₆ - alkyl,
_(Viii) C 1 ~C ₆ - alkylcarbonyl,
(Ix) _carboxy, C 1 -C ₄ - alkoxycarbonyl, carboxy _-C 1 -C ₆ - alkyl or _C 1 -C ₆ - alkoxycarbonyl _-C 1 -C ₆ - alkyl,
(X) hydroxy,
_(Xi) C 1 ~C ₆ - alkoxy,
(Xii) aryl _-C 1 -C ₄ - alkoxy, preferably benzyloxy,
_(Xiii) C 1 ~C ₆ - _{alkoxycarbonylamino,} C 1 -C ₆ - alkoxycarbonylamino _-C 1 -C ₆ - refers to an alkyl,
R7 represents C ₁ -C ₆ -alkylcarbonyl, preferably acetyl, propionyl, C ₁ -C ₆ -alkoxycarbonyl, preferably methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, and X and Y are oxygen or sulfur Indole derivatives, their tautomers, stereoisomers such as diastereomers and enantiomers and their physiologically tolerable salts.

  In the case where R represents an unsubstituted or substituted 2-, 3-, 4-, 5- or 6-pyridyl group and R1-R6 have the meanings given above, R7 in this case is an acetyl group or t- Do not represent a butyloxycarbonyl group.

  A further subject of the present invention is a compound of formula I

［式中、
Ｒはアミド窒素に直接的に結合された、
（ｉ）置換の６−キノリル、非置換又は置換の７−キノリル、その際、２−メチル−６−キノリルは例外とし、そうしてＸが硫黄原子である場合には、Ｒは非置換の６−キノリルであってもよい、
（ｉｉ）非置換又は置換の２−、３−、６−、７−及び８−ピリドピラジニル、
（ｉｉｉ）非置換又は置換の３−、４−、５−、６−及び７−インダゾリル
を意味し、
Ｒ１は非置換又は置換のアルキル−アリールを意味し、
Ｒ２は水素を意味し、
Ｒ３〜Ｒ６は
（ｘｉｖ）水素、
（ｘｖ）非置換又は置換のＣ_１〜Ｃ_６−アルキル、
（ｘｖｉ）非置換又は置換のＣ_３〜Ｃ_７−シクロアルキル、
（ｘｖｉｉ）アミノ、モノ−Ｃ_１〜Ｃ_４−アルキルアミノ、ジ−Ｃ_１〜Ｃ_４−アルキルアミノ、
（ｘｖｉｉｉ）ハロゲン、
（ｘｉｘ）１つ以上のフッ素原子で置換されたＣ_１〜Ｃ_４−アルキル、有利にはトリフルオロメチル基、
（ｘｘ）シアノ、直鎖状又は分枝鎖状のシアノ−Ｃ_１〜Ｃ_６−アルキル、
（ｘｘｉ）Ｃ_１〜Ｃ_６−アルキルカルボニル、
（ｘｘｉｉ）カルボキシル、Ｃ_１〜Ｃ_４−アルコキシカルボニル、カルボキシ−Ｃ_１〜Ｃ_６−アルキル又はＣ_１〜Ｃ_６−アルコキシカルボニル−Ｃ_１〜Ｃ_６−アルキル、
（ｘｘｉｉｉ）Ｃ_１〜Ｃ_６−アルコキシ、
（ｘｘｉｖ）アリール−Ｃ_１〜Ｃ_４−アルコキシ、有利にはベンジルオキシ、
（ｘｘｖ）Ｃ_１〜Ｃ_６−アルコキシカルボニルアミノ、Ｃ_１〜Ｃ_６−アルコキシカルボニルアミノ−Ｃ_１〜Ｃ_６−アルキル
を意味し、かつ
Ｒ７は水素を意味し、
Ｘ、Ｙは酸素又は硫黄を意味する］のインドール誘導体、それらの互変異性体、立体異性体、例えばジアステレオマー及びエナンチオマー並びにそれらの生理学的に認容性の塩である。

[Where:
R was directly attached to the amide nitrogen;
(I) substituted 6-quinolyl, unsubstituted or substituted 7-quinolyl, with the exception of 2-methyl-6-quinolyl, and when X is a sulfur atom, R is unsubstituted May be 6-quinolyl,
(Ii) unsubstituted or substituted 2-, 3-, 6-, 7- and 8-pyridopyrazinyl,
(Iii) means unsubstituted or substituted 3-, 4-, 5-, 6- and 7-indazolyl,
R1 means unsubstituted or substituted alkyl-aryl,
R2 means hydrogen,
R3 to R6 are (xiv) hydrogen,
(Xv) unsubstituted or substituted _C 1 -C ₆ - alkyl,
(Xvi) the unsubstituted or substituted _C 3 -C ₇ - cycloalkyl,
(Xvii) amino, _mono--C 1 -C ₄ - alkylamino, di _-C 1 -C ₄ - alkylamino,
(Xviii) halogen,
(Xix) C ₁ -C ₄ -alkyl substituted with one or more fluorine atoms, preferably a trifluoromethyl group,
(Xx) cyano, straight-chain or branched _cyano--C 1 -C ₆ - alkyl,
_(Xxi) C 1 ~C ₆ - alkylcarbonyl,
(Xxii) a _carboxyl, C 1 -C ₄ - alkoxycarbonyl, carboxy _-C 1 -C ₆ - alkyl or _C 1 -C ₆ - alkoxycarbonyl _-C 1 -C ₆ - alkyl,
_(Xxiii) C 1 ~C ₆ - alkoxy,
(Xxiv) aryl _-C 1 -C ₄ - alkoxy, preferably benzyloxy,
_(Xxv) C 1 ~C ₆ - _{alkoxycarbonylamino,} C 1 -C ₆ - alkoxycarbonylamino _-C 1 -C ₆ - refers to an alkyl, and R7 is a hydrogen,
Indole derivatives, their tautomers, stereoisomers such as diastereomers and enantiomers and physiologically acceptable salts thereof.

  The present invention is an improvement of the invention described in WO 02/10152. The indole derivatives include 2-methyl-6-quinolyl groups unsubstituted or substituted 2-, 3-, 6-, 7- and 8-pyridopyrazinyl, unsubstituted or substituted 3-, 4-, 5-, 6 It was confirmed that various tumor cell lines showed improved antiproliferative effects when substituted with-and 7-indazolyl.

  Furthermore, it has been confirmed that the compounds according to the invention have a potent cytotoxic effect which can be based on various mechanisms. The mechanism of the compounds according to the invention demonstrated in the above invention is based on inhibition of tubulin polymerization and inhibition of topoisomerase II. As a result, tumorigenic cells are arrested in the G2M phase. Furthermore, the compounds according to the invention induce apoptosis.

  Furthermore, it has been confirmed that the compounds according to the invention show an improved water solubility and thus an improved oral bioavailability.

  Furthermore, by introducing an acetyl group as the R7 group, the compounds according to the invention could show better compatibility with improved in vivo activity.

  It is desirable to use the substance classes described in the invention to give the possibility of medications that are smaller, long lasting, and compatible than using classical cytostatic drugs. In particular, it is desirable to avoid the disadvantageous resistance formation known from many antitumor agents. It is desirable that the enhanced action achieved with the indole derivatives according to the present invention make the consumption of pharmaceuticals more effective. Furthermore, it would be desirable to be able to extend treatment to the case of treatment resistance.

  In a preferred embodiment, in the indole derivatives of formula I, R1 represents 4-chlorobenzyl, R2 to R6 represent hydrogen, R represents a heterocycle, and R7 represents alkylcarbonyl or alkoxycarbonyl. To do.

  In a further advantageous embodiment, in the indole derivatives of the formula I, R represents unsubstituted 5-quinolyl, unsubstituted 6-quinolyl or unsubstituted 7-quinolyl and R7 represents acetyl or propionyl .

  In a further advantageous embodiment, in the indole derivatives of the formula I, R means unsubstituted 5-quinolyl, unsubstituted 6-quinolyl or unsubstituted 7-quinolyl and R7 is methoxycarbonyl, ethoxycarbonyl or Means propionoxycarbonyl.

  In the following, some concepts used in the specification and claims are defined.

  With respect to “heterocycle”, unless the concept is listed in detail in advance, pyrrole, furan, thiophene, pyrazole, thiazole, indole, oxazole, imidazole, isothiazole, isoxazole, 1,2,3-triazole, 1, 2,4-triazole, 1,2,4-oxadiazole, 1,3,4-oxadiazole, 1,2,5-thiadiazole, 1,3,4-thiadiazole, tetrazole, pyridine, pyrimidine, pyridazine, Pyrazine, benzofuran, indazole, carbazole, benzoxazole, benzimidazole, benzothiazole, benzotriazole, quinoline, cinnoline, quinoxaline, quinazoline, phthalazine, pyridopyrazine, 1,2,3-triazine, 1,2,4-triazine, 1, 3 5-triazine, represents purine, pteridine, acridine and phenanthridine.

The expression “alkyl”, within the scope of the present invention, includes hydrocarbons that may be acyclic, unsaturated or saturated, linear or branched. With respect to “alkyl”, the term “substituted” means within the scope of the present invention that a hydrogen radical is F, Cl, Br, I, CN, NH ₂ , NH— unless specifically defined in advance. Represents substituted by alkyl, NH-cycloalkyl, OH, O-alkyl, wherein the polysubstituted group is substituted at different or the same atom at multiple positions, for example at 2 or 3 positions. and are, for example _-CF 3, at three places on the same C atom as in the case of _-CH 2 CF _3, or at different positions as in the case of _{-CH (OH) -CH 2 -CH 2} -CHCl 2 Represents a group substituted at three positions. Multiple substitutions can be made with the same or different substituents.

The expression “alkyl-aryl” means C ₁ -C ₆ -alkyl-C ₆ -C ₁₄ -aryl and preferably C ₁ -C ₆ -alkyl-aryl. With reference to “alkyl-aryl” as well as “cycloalkyl”, within the scope of the present invention, unless otherwise specifically stated in advance, “mono- or polysubstituted” refers to one or more hydrogens of the ring system. atoms, F, Cl, Br, I , CN, NH 2, NH- alkyl, OH, O-alkyl, CF _{3, alkyl, C} 6 _{~C 10} - _{aryl, C} 6 _{~C 10} - aryl _-C 1 ~ Represents substitution on one or optionally different atoms in one or more places, for example 2, 3 or 4 places by C ₆ -alkyl and / or heterocyclyl (substituents optionally May itself be substituted). The polysubstitution is in this case carried out with the same or different substituents.

With respect to “heterocycle”, unless otherwise specifically stated in the scope of the present invention, “mono- or polysubstituted” means that one or more hydrogen atoms of the ring system are F, Cl , Br, I, nitro, _amino, C 1 -C ₆ - alkyl, preferably methyl, _mono--C 1 -C ₆ - alkylamino, di _-C 1 -C ₆ - alkylamino, _hydroxy, C 1 -C ₆ - alkoxy, benzyloxy, _carboxy, C 1 -C ₆ - _{alkoxycarbonyl,} C 1 -C ₆ - alkoxycarbonylamino or fluorine _C 1 monosubstituted or polysubstituted by -C ₆ - alkyl, preferably trifluoromethyl Methyl, C ₆ -C ₁₀ -aryl and / or C ₆ -C ₁₀ -aryl-C ₁ -C ₆ -alkyl in one or more places, for example in two, three or four places Indicates that it is substituted on one or optionally different atoms (the substituent may optionally be substituted itself). The polysubstitution is in this case carried out with the same or different substituents.

  If the compounds according to the invention of the general formula I have at least one asymmetric center, the compound is in racemic form, pure enantiomer and / or diastereomeric form or a mixture of enantiomers and / or diastereomers Can exist in the form of This mixture can be present in any mixing ratio of the stereoisomers. If possible, the compounds according to the invention can exist in tautomeric forms.

  For example, the compounds according to the invention of the general formula I which have one or more chiral centers and exist as racemates can be separated into optical isomers, ie enantiomers or diastereomers, by methods known per se. it can. This separation may be accomplished by column separation in a chiral phase or by recrystallization from an optically active solution, using optically active acids or bases, or by derivatization with optically active reagents such as optically active alcohols, and subsequent residues. It can be done by separation.

  A compound according to the invention of the general formula I, when it has a sufficiently acidic group, for example a carboxyl group, can be converted into a physiologically acceptable salt using inorganic and organic bases. Examples of the inorganic base include sodium hydroxide, potassium hydroxide and calcium hydroxide, and examples of the organic base include ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dibenzylethylenediamine and lysine. The stoichiometry of the salts produced of the compounds according to the invention is in this case an integer multiple of 1 or a non-integer multiple of 1.

  The compounds according to the invention of the general formula I can be converted into salts using inorganic and organic acids when they have a sufficiently basic group, for example a secondary or tertiary amine. Pharmaceutically acceptable salts of the compounds according to the invention according to general formula I are hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, carbonic acid, formic acid, acetic acid, trifluoroacetic acid , Oxalic acid, malonic acid, maleic acid, succinic acid, tartaric acid, glucose acid, malic acid, embonic acid, mandelic acid, fumaric acid, lactic acid, citric acid, glutamic acid or aspartic acid. The salts produced are in particular the hydrochloride, hydrobromide, sulfate, phosphate, methane sulfate, sulfoacetate, tosylate, carbonate, bicarbonate, formate, acetate, triflate, Oxalate, malonate, maleate, succinate, tartrate, malate, embonate, mandelate, fumarate, lactate, citrate and glutamate. The stoichiometry of the salts produced of the compounds according to the invention is in this case an integer multiple of 1 or a non-integer multiple of 1.

  Preference is likewise given to solvates, in particular hydrates, of the compounds according to the invention which can be obtained, for example, by crystallization from solvents or aqueous solutions. In this case, 1, 2, 3 or any number of solvent molecules or water molecules can be combined with the compounds according to the invention to form solvates and hydrates.

  It is known that chemicals form solids that exist in a variety of ordered states expressed as polymorphs or transformations. The various transformations of polymorphic substances differ significantly in their physical properties. The compounds according to the invention of the general formula I can exist in various polymorphic forms, in which the given modification is metastable.

  Both the compounds of formula I and their salts are biologically active. The compounds of formula I can be administered in free form or as a salt with a physiologically acceptable acid or base.

  Administration of the compounds of the general formula may be effected orally, rectally, buccal (sublingual), parenterally (eg subcutaneously, intramuscularly, enterally or intravenously), topically or transdermally.

  Furthermore, the present invention relates to at least one compound of the formula I or salts thereof with physiologically tolerable inorganic or organic acids and optionally pharmaceutically usable carrier substances and / or diluent substances or It relates to a medicinal product having an auxiliary content.

  These medicaments are used for the treatment of tumor diseases, particularly in tumor diseases having drug resistance to another agent and / or in tumor diseases with metastatic carcinoma.

  Applicable forms are, for example, tablets, dragees, capsules, drops or ampoules, suppositories, salves, inhalable powder preparations, suspensions, creams and ointments.

  The compounds according to the invention may be finely divided, for example dispersed in a nanoparticle composition.

The therapeutically useful properties found are related in particular to the following advantages:
The compounds according to the invention are distinguished by strong and highly proliferative activity properties;
The compounds according to the invention inhibit tubulin polymerization;
The compounds according to the invention inhibit topoisomerase II;
The compounds according to the invention arrest dividing cells in the G2 / M phase;
The compounds according to the invention induce apoptosis;
The compounds according to the invention are distinguished by potent antitumor in vivo activity and even improved compatibility;
The compounds of formula I according to the invention are effective in vitro in mdr-resistant cell lines as opposed to paclitaxel, vincristine, doxorubicin or etoposide.

Most preferred compounds have the following options, represented by general formula I:
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-pyrido [2,3-b] pyrazin-7-yl-acetamide (1),
2- [1- (4-chloro-benzyl) -1H-indol-3-yl] -N- (1H-indazol-5-yl) -2-oxo-acetamide (4),
N- {2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -N-quinolin-6-yl-acetamide (2),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-methyl ester (3),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-ethyl ester (5),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-propyl ester (6),
N- {2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -N-quinolin-6-yl-propionamide (7),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -pyridin-4-yl-carbamic acid-ethyl ester (8),
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -N-quinolin-6-yl-2-thio-acetamide (11)
It is a compound that matches

  Compounds (1), (4) and (11) are compounds in which the group R7 means hydrogen. Compounds (2), (3), (5) and (6) to (8) have an alkylcarbonyl group or an alkoxycarbonyl group as the group R7.

The following compounds (9), (10), (12), (13), (14) and (15) are compounds investigated for the purpose of comparison. Compounds (9), (10), (14) and (15) are known from the prior art. Compound (9) is described in Applicant's WO02 / 10152, Compound (10) is described in WO03 / 022280, Compound (13) is included in the claims of WO02 / 08225A, Compound ( 12), (14) and (15) are included in the claims of WO99 / 51224 A1 and WO01 / 22954.
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -N- (2-methyl-quinolin-6-yl) -2-oxo-acetamide (9),
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-quinolin-6-yl-acetamide (10),
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-quinolin-8-yl-acetamide (12),
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -N-isoquinolin-5-yl-2-oxo-acetamide (13),
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-pyridin-4-yl-acetamide (14),
2- [1- (4-Fluoro-benzyl) -1H-indol-3-yl] -N- (2-methyl-quinolin-8-yl) -2-oxo-acetamide (15)
Compounds of general formulas Ia and Ib of the reaction scheme are obtained by the following reaction scheme 1:

  Compounds of general formula Ic where X is S can be prepared according to Scheme 2:

  The compounds of general formula Ic and Y being S are obtained according to methods known from the literature (W.-D. Malmberg et al. Liebigs Ann. Chem. 10, 1983; 1649-1711).

  These starting compounds II, III and IV are commercially available or can be prepared by methods known per se. Starting materials II, III and IV are important intermediate compounds for the preparation of the indole derivatives according to the invention of formula I.

For the preparation of starting compounds and target compounds, for example, the following standard studies of organic synthesis are pointed out. These contents should be the disclosure elements of the present application with this:
・ Houben-Weyl, Volume E7a (Part 1), 290-492, 571-740,
Houben-Weyl, Volume E7a (Part 2), 119-156, 205-686, 157-204,
Monographie, "Heterosyclic Compounds" (Elderfield),
Volume 1, pages 119-207, pages 397-616,
Volume 3, pages 1 to 274,
Volume 6, pages 101-135, pages 234-323,
・ Monographie "Comprehensive Organic Chemistry" (SDBarton, WDOllis)
Vol. 4, pp. 155-204, pp. 205-232, 493-564 The solvents and auxiliaries to be used optionally and the reaction parameters to be applied, for example the reaction temperature and reaction time, are based on their expertise by those skilled in the art. It is known.

The following compounds were synthesized according to the general instructions for Step 1, Step 2 and Step 3, which are the basis of Synthetic Schemes 1 and 2. Each chemical name is clear from the following summary. Analytical characterization of the compounds according to the invention was carried out by their melting point or ¹ H-NMR spectroscopy and / or mass spectrometry.

  The chemicals and solvents used were either commercially available or synthesized from common sources (Acros, Avocado, Aldrich, Fluka, Lancaster, Maybridge, Merck, Sigma, TCI, etc.).

  The present invention will be described in detail with reference to the following examples, but the present invention is not limited to these examples.

Example
Example 1 (reaction according to reaction scheme 1, first stage):
Preparation of 1- (4-chlorobenzyl) -indole 1.32 g of sodium hydride (0.055 mol, mineral oil suspension) in a mixture of 50 ml of dimethyl sulfoxide, 5.86 g (0.05 mol) A solution of indole in 25 ml of DMSO is added. Heat at 60 ° C. for 1.3 hours, then cool and add 17.7 g (0.11 mol) of 4-chloro-benzyl chloride dropwise. The solution is heated to 60 ° C., allowed to stand overnight and poured into 200 ml of water with further stirring. Extract multiple times with a total of 75 ml of CH ₂ Cl ₂ , dry the organic phase over anhydrous sodium sulfate, filter and concentrate the filtrate in vacuo.

Yield: 11.5 g (95% of theory)
Example 2 (reaction according to the second stage of reaction formula 1):
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-pyrido [2,3-b] pyrazin-7-yl-acetamide (1)
To a solution of 1.12 ml oxalyl chloride in 50 ml ether, 10.2 g (10.7 mmol) 1- (4-chlorobenzyl) -indole in 200 ml ether at 0 ° C. under nitrogen. Add the dissolved solution dropwise. Heat under reflux for 2 hours and subsequently evaporate the solvent. Then 30 ml of DMF is added to the residue, 1.93 g (13.9 mmol) of potassium carbonate is added, the suspension is cooled to 0 ° C. and 1.57 g (10.7 mmol) of amine is added. A solution of the components in 10 ml DMF is added dropwise. Stir overnight at room temperature. The reaction mixture is finally introduced into ice water with stirring and the resulting precipitate is separated off with suction. The crude product obtained is chromatographed on 100 g of silica gel with n-heptane / acetic acid = 4: 1.

Yield: 3.23 g (68.0%)
Freezing point: 250 ° C
¹ H-NMR (DMSO-D6) δ = 11.56 (s, 1H), 9.53 (d, 1H), 9.12 (s, 1H), 9.09 (d, 1H), 9.04 (S, 1H), 8.32 (d, 1H), 7.6 (d, 1H), 7.40 (d, 2H), 7.35 (m, 3H), 7.32 (m, 2H) , 5.64 (s, 2H) ppm
Example 3 (reaction according to the third stage (a) of reaction formula 1):
N- {2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -N-quinolin-6-yl-acetamide (2)
6.0 g (13.6 mmol) 2- [1- (4-chlorobenzyl) -1H-indol-3-yl] -2-oxo-N-quinolin-6-yl-acetamide in 60 ml DMF To the dissolved and stirred solution, 0.833 g (6.82 mmol) of DMAP, 1.38 g (13.6 mmol) of triethylamine and 13.9 g (136 mmol) of acetic anhydride are added under nitrogen. The reaction mixture is stirred at room temperature for 10 minutes and then 200 ml of ethyl acetate is injected. After adding 300 ml of water, the mixture is shaken in a separatory funnel and then both phases are allowed to separate. Settling begins after 20 minutes. The pale yellow crystals are filtered off and dried at 60 ° C. under vacuum.

Yield: 4.04 g (61.5%)
Freezing point: 122.9 ° C
¹ H-NMR (600 MHz, DMSO-d6) δ = 9.02 (d, 1H), 8.54 (s, 1H), 8.44 (d, 1H), 8.21 (d, 1H), 8 .17 (d, 1H), 8.10 (m, 1H), 7.88 (m, 1H), 7.65 (m, 1H), 7.58 (m, 1H), 7.44 (d, 2H), 7.33 (d, 2H), 7.28 (m, 2H), 5.60 (s, 2H), 2.15 (s, 3H)
MS (ESI) m / z 482.1 (MH ^<+> ), (theoretical 481.94)
Example 4 (reaction according to the third stage (a) of reaction formula 1):
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-methyl ester (3)
10.0 g (22.7 mmol) of 2- [1- (4-chlorobenzyl) -1H-indol-3-yl] -2-oxo-N-quinolin-6-yl-acetamide in 500 ml of anhydrous THF 930.2 mg (27.3 mmol) of NaH (also as a 60% dispersion in mineral oil) is added under nitrogen to the stirred solution. The solution is stirred at 0 ° C. until a yellow precipitate precipitates and then stirred for a further 15 minutes. Then 2.58 g (27.3 mmol) of methyl chloroformate are added dropwise at a temperature below + 5 ° C. The reaction is monitored by thin layer chromatography (eluent: n-heptane / ethyl acetate 1/1 RF = 0.11). The reaction mixture is poured into water and extracted with ethyl acetate, the organic phase is washed with saturated sodium chloride solution and dried over anhydrous MgSO ₄ . Evaporation of the solvent gives a crude product, which is purified by column chromatography (n-heptane / acetone 2/1) to give compound 3. Since compound 3 was low in impurities by thin layer chromatography, it can be removed by stirring crude compound 3 with acetone for 1 hour. Filtration gives compound 3 as pale yellow crystals.

Yield: 3.0 g (26.5%)
Freezing point: 178.5 ° C
¹ H-NMR (600 MHz, DMSO-d6) δ = 9.02 (d, 1H), 8.58 (s, 1H), 8.47 (d, 1H), 8.17 (m, 3H), 7 .84 (m, 1H), 7.63 (m, 2H), 7.44 (d, 2H), 7.34 (m, 4H), 5.60 (s, 2H), 3.65 (s, 3H)
MS (ESI) m / z 498.2 (MH ^<+> ), (theoretical 497.94)
Example 5 (reaction according to the third stage (b) of reaction formula 2):
Preparation of 2- [1- (4-chloro-benzyl) -1H-indol-3-yl] -N-quinolin-6-yl-2-thioxo-acetamide (11) 4.00 g (9.1 mmol) A suspension of 2- [1- (4-chlorobenzyl) -1H-indol-3-yl] -2-oxo-N-quinolin-6-yl-acetamide in 200 ml of toluene was added to nitrogen. Below, 3.68 g (9.1 mmol) of 2,4-bis- (4-methoxyphenyl) -1,3-dithia-2,4-diphosphetane-2,4-disulfide were mixed, followed by 75 ° C. For 3 hours. From the reaction solution, the residue formed at elevated temperature is filtered off and subsequently washed with 100 ml of methylene chloride. The filtrate is concentrated in vacuo and the residue is subjected to flash silica gel chromatography (eluent: methylene chloride / methanol 99: 1). The product fraction is filtered over flash silica gel after removing the solvent again in vacuo (eluent: n-heptane / ethyl acetate 1: 1).

Yield: 0.46 g (11% of theory)
ESI-MS m / e 456.1 (MH ⁺ ), (theoretical value 455.97)
^{1 H-MNR (DMSO-D6} ) δ = 10.89 (s, 1H), 8.8 (s, 1H), 8.75 (s, 1H), 8.55 (s, 1H), 8.12 (D, 1H), 8.35 (d, 1H), 8.0 (d, 1H), 7.93 (d, 1H), 7.63 (d, 1H), 7.50 (m, 1H) , 7.4 (m, 3H), 7.3 (m, 3H), 5.6 (s, 2H) ppm
The following compounds of formula I were synthesized analogously to the synthetic route of Scheme 1 and according to Examples 2 and 3.

Example 6:
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -N- (1H-indazol-5-yl) -2-oxo-acetamide (4)
Freezing point: 203 ° C
¹ H-NMR (DMSO-D6) δ = 13.02 (s, 1H), 10.7 (s, 1H), 9.04 (s, 1H), 8.48 (s, 1H), 8.42 (D, 1H), 8.06 (s, 1H), 7.73 (d, 1H), 7.6 (d, 1H), 7.55 (d, 1H), 7.40 (d, 2H) , 7.28-7.35 (m, 4H), 5.63 (s, 2H) ppm
Example 7:
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-ethyl ester (5)
Freezing point: 199 ° C
¹ H-NMR (600 MHz, DMSO-d6) δ = 9.02 (m, 1H), 8.60 (s, 1H), 8.48 (d, 1H), 8.15 (m, 3H), 7 .83 (m, 1H), 7.63 (m, sH), 7.43 (d, 2H), 7.32 (m, 4H), 5.60 (s, 2H), 4.15 (q, 2H), 0.95 (t, 3H)
MS (ESI) m / z 514.2, 512.1 (MH ^<+> ), (theoretical value 511.97)
Example 8:
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-propyl ester (6)
Freezing point: 164 ° C
^{1 H-NMR (600MHz, DMSO} -d6) δ = 9.02 (m, 1H), 8.60 (s, 1H), 8.48 (d, 1H), 8.17 (m, 3H), 7 .84 (m, 1H), 7.63 (m, 2H), 7.43 (d, 2H), 7.33 (m, 4H), 5.61 (s, 2H), 4.03 (t, 2H), 1.32 (m, 2H), 0.56 (t, 3H)
Example 9:
N- {2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -N-quinolin-6-yl-propionamide (7)
¹ H-NMR (600 MHz, DMSO-d6) δ = 9.03 (m, 1H), 8.52 (s, 1H), 8.45 (d, 1H), 8.23 (d, 2H), 8 .18 (d, 1H), 8.13 (m, 1H), 7.88 (m, 1H), 7.65 (m, 1H), 7.58 (m, 1H), 7.45 (d, 2H), 7.30 (m, 4H), 5.59 (s, 2H), 2.61 (q, 3H), 0.88 (t, 3H)
Example 10:
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -pyridin-4-yl-carbamic acid ethyl ester (8)
Freezing point: 62 ° C
¹ H-NMR (500 MHz, DMSO-d6) δ = 8.74 (m, 2H), 8.52 (s, 1H), 8.12 (m, 1H), 7.60 (m, 1H), 7 .55 (m, 2H), 7.40 (m, 2H), 7.30 (m, 4H), 5.57 (s, 2H), 4.10 (q, 2H), 0.95 (t, 3H)
Example 11 (comparative substance):
Preparation of 2- [1- (4-chloro-benzyl) -1H-indol-3-yl] -N- (2-methyl-quinolin-6-yl) -2-oxo-acetamide (9) Yield: 14. 8g (77.3% of theoretical value)
Freezing point: 182-185 ° C
¹ H-NMR (CDCl ₃ ) δ = 9.58 (s, 1H), 9.12 (s, 1H), 8.5 (s, 1H), 8.41 (s, 1H), 8.05 ( t, 2H), 7.78 (d, 1H), 7.4 (dd, 1H), 7.32 (m, 4H), 7.26 (s, 1H), 7.15 (d, 1H), 5.38 (s, 2H), 2.73 (s, 3H) ppm
Example 12 (comparative substance):
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-quinolin-6-yl-acetamide (10)
Freezing point: 200 ° C
¹ H-NMR (DMSO-D ₆ ) δ = 11.5 (s, 1H), 9.05 (s, 1H), 8.85 (s, 1H), 8.66 (s, 1H), 8. 32 (d, 2H), 8.12 (d, 1H), 8.03 (d, 1H), 7.63 (d, 1H), 7.53 (dd, 1H), 7.42 (d, 2H) ), 7.30-7.38 (m, 4H), 5.63 (s, 2H) ppm
Example 13 (comparative substance):
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-quinolin-8-yl-acetamide (12)
Freezing point: 178 ° C
Example 14 (comparative substance):
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -N-isoquinolin-5-yl-2-oxo-acetamide (13)
Freezing point: 239-241 ° C
Example 15 (comparative substance):
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-pyridin-4-yl-acetamide (14)
Freezing point: 264 ° C
Example 16 (comparative substance):
2- [1- (4-Fluoro-benzyl) -1H-indol-3-yl] -N- (2-methyl-quinolin-8-yl) -2-oxo-acetamide (15)
Freezing point: 200-202 ° C
Biological effects of the compounds according to the invention In vitro and in vivo tests on selected tumor models yielded the following pharmacological activity.

Example 17: were investigated for their antiproliferative activity with respect to an established tumor cell line in proliferation tests The antiproliferative agents 1,2,4,9,11,12,13 and 15 relating to a variety of tumor cell lines (DA Scuderio et al. Cancer Res. 1988, 48, 4827-4833). The test used determines cellular dehydrogenase activity and allows measurement of cell activity and indirectly of cell number. The cell lines used were human cervical cancer cell line KB / HeLa (ATCC CCL17), ovarian adenocarcinoma cell line SKOV-3 (ATCC HTB77), human glioma cell line SF-268 (NCI 503138) and lung cancer cell line NCI. -H460 (NCI 503473).

TABLE 1 Growth inhibition of substances according to the invention in human tumor cell lines in the XTT cytotoxicity test This result shows that according to Examples 1, 2, 4 and 11, the growth of selected tumor cell lines is very strong. It shows that it was inhibited.

Example 18: Anti-proliferative effect on MDR tumor cell lines For further characterization, substances 1, 2, 4 and 11 were investigated against multi-drug resistant cell lines compared to non-resistant wild type cell lines. did.

  The cell lines investigated were mouse L1210, acute myeloid leukemia cell line LT12 and resistant cell lines L1210 / mdr and LT12 / mdr. In addition, the mouse P388 cell line (methyl-cholanthrene-induced lymphoid tumor) and doxorubicin-resistant P388 were considered as test systems.

  The results are summarized in Table 2 below:

Table 2: Inhibitory effects of substances in human tumor cell lines in the XTT proliferation test Substances 1, 2, 4 and 11 show very potent inhibitory effects in all tested cell lines, but classical tubulin In the case of substances exhibiting an inhibitory action, such as paclitaxel or vincristine, and in the case of topoisomerase II inhibitors (doxorubicin, mitoxantrone, etoposide), at least a drastic reduction of the action is observed in the MDR1-resistant cell line.

Example 19: Inhibitors of Tubulin Polymerization 1, 4, 9, 11, 12, 13 and 15 were tested for inhibition of bovine tubulin polymerization in an in vitro test (DM Bollag et al. Cancer Res. 1995, 55, 2325-2333). In this test, tubulin purified by a polymerization and depolymerization cycle was used, and this tubulin was polymerized by addition of GTP and heating. Table 3 shows the EC ₅₀ values for inhibition of tubulin polymerization by 30% related protein (MAP).

Table 3: Inhibition of tubulin polymerization. Mean values from two independent tests The results (see Table 3) show a very potent inhibitory action of substances 1, 4, 9 and 11 on tubulin polymerization, whereas compounds 12, 13 and 15 are effective. It shows that does not demonstrate.

Example 20 Topoisomerase II Inhibitor 1 was investigated for inhibition of topoisomerase II in two different in vitro tests.

A kDNA assay for testing topoisomerase II activity In the assay described by Arimondo (Anti-Cancer Drug design 2000, 15 (6), 413-421), kDNA is treated with human DNA topoisomerase II in the absence or presence of the test compound. The test of compound 1 according to the invention was carried out in this case at three different concentrations (100 μM, 31.6 μM and 10 μM). For comparison, positive control and reference compounds m-amsacrine (m-Amsa), paclitaxel (Taxol) and vincristine were used at a concentration of 100 μM, respectively.

Performing the assay:
1 μL of test material (concentrated 20-fold in 100% DMSO) to 2 μL of 10 × assay buffer, 1 μL of kDNA (200 ng), 0.5 μL of human topoisomerase II (1 unit) And 15.5 μL of H ₂ O were added by pipette and mixed.

The reaction batch is placed in a heating block preheated to 37 ° C. and incubated at 37 ° C. for 10 minutes. Incubation is stopped by adding 4 μL of 5 × stop buffer followed by extraction of material with CIA. 20 μL of the supernatant is then loaded onto a 1% agarose gel containing 0.25 μg / mL ethidium bromide and allowed to separate at 100 V for 1 hour. Finally, the gel is photographed under UV excitation (see FIG. 1). Quantification of the inhibition of decatenation of kDNA performed in GelPro ^(TM) Analyzer Software (see Figure 2).

PRYG relaxation assay for testing of topoisomerase II activity:
This relaxation assay could further demonstrate the inhibitory properties of the compounds according to the invention on topoisomerase II. The test of compound 1 according to the invention was carried out in this case at three different concentrations (100 μM, 31.6 μM and 10 μM). For comparison, the reference compounds m-amsacrine, paclitaxel (Taxol) and vincristine were used at concentrations of 316 μM and 100 μM.

In this case, the implementation is as follows:
1 μL of test material (concentrated 20-fold in 100% DMSO) was added to 2 μL of 10 × assay buffer, 0.5 μL of pRYG DNA (125 ng), 0.5 μL of human topoisomerase II ( 1 unit) and 16 μL of H ₂ O were added by pipette and mixed. The reaction batch is placed in a heating block preheated to 37 ° C and incubated at 37 ° C for 30 minutes. Incubation is stopped by adding 4 μL of 5 × stop buffer. The 10 μL batch is then loaded onto a 1.2% agarose gel containing 0.25 μg / mL ethidium bromide and allowed to separate at 100 V for 2.5 hours. Finally, the gel is photographed under UV excitation (see FIG. 3). Quantification of the inhibition of pRYG relaxation performed in GelPro ^(TM) Analyzer Software (see FIG. 4).

  Overall, it can be confirmed that high inhibition of topoisomerase II could be detected in both assays for compound 1 according to the invention. The results for compound 1 are comparable to the inhibition values of the topoisomerase II inhibitor m-amsacrine. For paclitaxel and vincristine, no inhibitory effects were observed as expected in both assays.

Example 21: Cell cycle analysis The cell cycle includes cell generation from one cell generation to the next.

  Between the resting phase (G0) and the synthetic preparation phase (G1), the cells have a polyploid chromosome set (2c). In the synthesis phase (S), the amount of DNA increases due to replication. The S phase ends when it reaches the preparatory division (G2M), during which time the cells have duplicated chromosomal mass (4c) and double the DNA content. In the subsequent short mitotic phase (M), the replicated chromosomes are simultaneously distributed to the two daughter cells, each again showing multiple DNA content and being in the G01 phase so that the cell cycle can be started anew.

  For cell cycle analysis, KB / HeLa cells were treated with test substances at various concentrations (0.1-1000 nM) for 24 hours at 37 ° C.

The percentage of cells arrested in the G2 / M phase of the cell cycle after treatment with a reference substance or selected test substance is shown in Table 4 below. The results were evaluated with specific analysis software (ModFit ^™) .

Table 4: Inhibitory concentrations for 50% cell proportion in G2 / M phase Compounds 1, 2, 4 and 11 according to the present invention have comparable activity to the reference compounds paclitaxel and mitoxantrone.

Example 22: Detection of apoptosis CDD ^plus nucleosome ELISA test:
Nuclear fragmentation is a result after the apoptotic process. The changes to be observed in this case are due to the DNA strands degraded by the endonuclease and the resulting fragmentation into nucleosome particles.

  For detection of nucleosome particles, the CDDplus nucleosome ELISA test described by Roche Molecular Biochemicals was used. For this, compounds 1 and 2 were investigated in U-937 cell line at various concentrations (1 nM to 10 μM; treated for 24 hours). (See FIGS. 5 and 6).

  For compounds 1 and 2, in this case, a concentration-dependent increase in nucleosomes could be observed in the cell lysate. There is no significant increase in cell culture supernatant, which provides evidence for apoptotic cell death after treatment with compounds 1 and 2.

Example 23: Verification of saturation solubility in water for compounds according to the invention The determination of saturation solubility in water for compounds 1, 2, 10 and 14 was carried out as described below. Up to 1% DMSO was added to improve material dissolution and sample wetting. The HPLC-UV method was used to investigate the content. The results are summarized in Table 5 below:

Table 5: Saturated Solubility of Compounds 1, 2, 10 and 14 Compounds 1 and 2 according to the invention are superior in improved water solubility compared to compounds 10 and 14.

Example 24: was tested in vivo activity present invention by the compounds 2 in vivo activity and compatibility with compared to material 10 and 14 human xenograft models (melanoma, MEXF462). The results are summarized in Table 6 below:
In vivo activity of compounds 2, 10 and 14 (melanoma; MEXF462)

Table 6: In vivo activity of compounds 2, 10 and 14 (melanoma; MEXF462)
For compound 2, complete tumor remission was further observed in the xenograft model with very good fit in the treated animals (no deaths). For compounds 10 and 14, comparable anti-tumor effects were observed, but the compatibility was not good.

Example 25: In vivo activity The in vivo activity and suitability of compound 2 according to the invention was tested in another human xenograft model (mammary gland, MEXF857) compared to substance 10. The results are shown in the following table:
Effect of compounds 2 and 10 on breast cancer MAXF857

Table 7: In vivo activity of compounds 2 and 10 (mammary gland; MAXF857)
Equivalent anti-tumor effects were observed for compounds 2 and 10, but the suitability of substance 10 (2/6 mice died) was substantially worse compared to compound 2.

FIG. 1 is a photograph of a gel showing the results of a kDNA assay. FIG. 2 is a graph showing the results of inhibition of kDNA dissociation circles. FIG. 3 is a photograph of a gel showing the results of the pRYG relaxation assay. FIG. 4 is a graph showing the results of inhibition of pRYG DNA relaxation. FIG. 5 is a graph showing the results of CCD ^plus nucleosome ELISA test with Compound 1. FIG. 6 is a graph showing the results of a CCD ^plus nucleosome ELISA test with Compound 2.

Claims (13)

Formula I

[Where:
R is saturated with one or more heteroatoms selected from the group of N, O and S, unsaturated or aromatic, coupled directly to the amide nitrogen, substituted or unsubstituted C ₂ -C 14 _- Complex Represents a ring,
R1 means unsubstituted or substituted alkyl-aryl,
R2 is i) hydrogen,
ii) unsubstituted or substituted _C 1 -C ₆ - refers to an alkyl,
R3 to R6 are i) hydrogen,
ii) unsubstituted or substituted _C 1 -C ₆ - alkyl,
iii) unsubstituted or substituted _C 3 -C ₇ - cycloalkyl,
iv) amino, _mono--C 1 -C ₄ - alkylamino, di _-C 1 -C ₄ - alkylamino,
v) halogen,
vi) 1 or more _C 1 are substituted with fluorine atoms -C ₄ - alkyl, preferably trifluoromethyl group,
vii) cyano, straight-chain or branched _cyano--C 1 -C ₆ - alkyl,
_viii) C 1 ~C ₆ - alkylcarbonyl,
ix) _carboxyl, C 1 -C ₄ - alkoxycarbonyl, carboxy _-C 1 -C ₆ - alkyl or _C 1 -C ₆ - alkoxycarbonyl _-C 1 -C ₆ - alkyl,
x) hydroxy,
_xi) C 1 ~C ₆ - alkoxy,
xii) aryl _-C 1 -C ₄ - alkoxy, preferably benzyloxy,
_xiii) C 1 ~C ₆ - _{alkoxycarbonylamino,} C 1 -C ₆ - alkoxycarbonylamino _-C 1 -C ₆ - refers to an alkyl,
R7 is _C 1 -C ₆ - alkylcarbonyl or _C 1 -C ₆ - means alkoxycarbonyl, and X, Y is meant oxygen or sulfur, Here, R is an unsubstituted or substituted 2-, 3- , 4-, 5- or 6-pyridyl group, and R7 does not represent an acetyl group or a t-butyloxycarbonyl group when R1 to R6 have the above-mentioned meanings], an indole derivative thereof, Mutants, stereoisomers such as diastereomers and enantiomers and physiologically acceptable salts thereof. R is i) unsubstituted or substituted 5-, 6-, 7-quinolyl,
ii) unsubstituted or substituted 2-, 3-, 6-, 7- and 8-pyridopyrazinyl,
iii) unsubstituted or substituted 3-, 4-, 5-, 6- and 7-indazolyl,
iv) unsubstituted or substituted 2-, 3-, 4-, 5- and 6-pyridyl,
v) unsubstituted or substituted 3-, 4- and 5-isoxazolyl,
vi) Indole derivatives according to claim 1, meaning unsubstituted or substituted 3-, 4- and 5-isothiazolyl.   The indole derivative according to claim 1 or 2, wherein R7 is methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, acetyl or propionyl. A compound of general formula I has the following compound:
N- {2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -N-quinolin-6-yl-acetamide (2),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-methyl ester (3),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-ethyl ester (5),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -quinolin-6-yl-carbamic acid-propyl ester (6),
N- {2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -N-quinolin-6-yl-propionamide (7),
{2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-acetyl} -pyridin-4-yl-carbamic acid-ethyl ester (8)
The indole derivative according to any one of claims 1 to 3, which is selected from the group consisting of: Represented by Formula I,
R was directly attached to the amide nitrogen,
(I) substituted 6-quinolyl, unsubstituted or substituted 7-quinolyl, with the exception of 2-methyl-6-quinolyl, and when X is a sulfur atom, R is unsubstituted May be 6-quinolyl,
(Ii) unsubstituted or substituted 2-, 3-, 6-, 7- and 8-pyridopyrazinyl,
(Iii) means unsubstituted or substituted 3-, 4-, 5-, 6- and 7-indazolyl,
R1 means unsubstituted or substituted alkyl-aryl,
R2 means hydrogen,
R3 to R6 are (i) hydrogen,
(Ii) unsubstituted or substituted _C 1 -C ₆ - alkyl,
(Iii) unsubstituted or substituted _C 3 -C ₇ - cycloalkyl,
(Iv) amino, _mono--C 1 -C ₄ - alkylamino, di _-C 1 -C ₄ - alkylamino,
(V) halogen,
(Vi) 1 or more _C 1 are substituted with fluorine atoms -C ₄ - alkyl, preferably trifluoromethyl group,
(Vii) cyano, straight-chain or branched _cyano--C 1 -C ₆ - alkyl,
_(Viii) C 1 ~C ₆ - alkylcarbonyl,
(Ix) _carboxy, C 1 -C ₄ - alkoxycarbonyl, carboxy _-C 1 -C ₆ - alkyl or _C 1 -C ₆ - alkoxycarbonyl _-C 1 -C ₆ - alkyl,
_(X) C 1 ~C ₆ - alkoxy,
(Xii) aryl _-C 1 -C ₄ - alkoxy, preferably benzyloxy,
_(Xii) C 1 ~C ₆ - _{alkoxycarbonylamino,} C 1 -C ₆ - alkoxycarbonylamino _-C 1 -C ₆ - refers to an alkyl, and R7 is a hydrogen,
X, Y are indole derivatives meaning oxygen or sulfur, their tautomers, stereoisomers such as diastereomers and enantiomers and their physiologically acceptable salts. The compound of formula I is the following compound:
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -N-quinolin-6-yl-2-thioxo-acetamide (11),
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -2-oxo-N-pyrido [2,3-b] pyrazin-7-yl-acetamide (1),
2- [1- (4-Chloro-benzyl) -1H-indol-3-yl] -N- (1H-indazol-5-yl) -2-oxo-acetamide (4)
6. The indole derivative according to claim 5, selected from the group of   The indole derivative according to any one of claims 1 to 6, wherein R 1 represents 4-chlorobenzyl, R 2 to R 6 represent hydrogen, and X and Y represent oxygen or sulfur.   A pharmaceutical comprising at least one indole derivative according to any one of claims 1 to 7.   9. The pharmaceutical product according to claim 8, comprising an indole derivative in the form of microparticles or nanoparticles.   Tablets, dragees, capsules, infusions or ampoules, suppositories, salves, inhalable powder formulations containing indole derivatives and pharmaceutically usable carrier and / or diluents and auxiliaries 10. A pharmaceutical product according to claim 8 or 9, in the form of a suspension, cream or ointment.   Use of the indole derivative according to any one of claims 1 to 6 for the manufacture of a medicament for the treatment of tumor diseases.   12. Use according to claim 11 for the treatment of tumor diseases having drug resistance to another agent.   12. Use according to claim 11 for the treatment of a tumor disease with metastatic carcinoma.

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