WO2013079754A1

WO2013079754A1 - Fluorinated indolenines for use in the treatment of cancer

Info

Publication number: WO2013079754A1
Application number: PCT/ES2012/070830
Authority: WO
Inventors: Sara Preciado Gallego; Alba PÉREZ PERARNAU; Joan Gil Santano; Fernando Albericio Palomera; Rodolfo LAVILLA GRÍFOLS
Original assignee: Universitat De Barcelona; Fundació Privada Institut D'investigació Biomèdica De Bellvitge; Fundació Privada Institut De Recerca Biomèdica
Priority date: 2011-11-28
Filing date: 2012-11-27
Publication date: 2013-06-06
Also published as: ES2410704A8; ES2410704A1; ES2410704B1

Abstract

The compounds of formula (I), or the pharmaceutically acceptable salts thereof or the stereoisomers or mixture of stereoisomers thereof, in which: R₁, R₂, R_4, R₅ and R₇ are independently selected from the group consisting of H, Cl, C(=O)O(C₁-C₄)alkyl, O(C₁-C₄)alkyl; and (C₁-C₄)alkyl; R₃ is selected from the group consisting of H₁, Cl, C(=O)O(C₁-C₄)alkyl, O(C₁-C₄)alkyl, (C₁-C₄)alkyl, pyrrolidinyl-1-carbonyl, and morpholin-1-carbonyl; R₆ is selected from the group consisting of H and (C₁-C₄)alkyl; and R₈ is selected from the group consisting of H, F, Br, (C₁-C₄)alkyl, and phenyl; said compounds inhibit the cellular proliferation of tumour cells independently of the p53 protein and are also able to induce apoptosis in various tumour cells independently of the p53 protein, which makes them useful for the treatment of various types of cancer.

Description

Fluorinated Indolenins useful for cancer treatment

The present invention is related to new compounds of

indolenin, pharmaceutical compositions containing them and their use for the treatment of cancer.

STATE OF THE TECHNIQUE

Cancer is a heterogeneous disease characterized by the accumulation of tumor cells, which can cause death in both animals and humans. Conventional methods for cancer treatment include surgical treatments, agent administration

chemotherapeutic agents, and more recently immune response based treatments, which involve the administration of an antibody or an antibody fragment that can be conjugated to a unit

therapy. However, so far, such treatments have had limited success.

Chemotherapy, despite all its limitations, is still today one of the most widespread methods for the treatment of different types of cancer. Therefore, the development of new antitumor therapies of general applicability is one of the main objectives of medical chemistry.

The inability of chemical agents to distinguish between normal rapidly dividing cells and tumor cells can lead to a depression of the patient's immune system. This has been considered one of the main problems associated with chemotherapy, as well as the resistance mechanisms developed by cancer cells, which include the inactivation of the p53 pathway. Although the majority of drugs currently used in therapy induce apoptosis in these cells, at least partially, through the activation of the p53 pathway, the p53 gene is mutated in half of the tumors analyzed, demonstrating its importance in development of cancer Great efforts are being made to improve antitumor treatments, trying to get active and selective compounds that they can act independently of the p53 pathway, to administer to patients with incipient and recurrent cancer or with metastases.

However, despite the efforts made so far, there is currently no curative therapy for most tumors, so there is still a need to find effective antitumor agents. In particular, it would be of great interest to find antitumor therapies that can act independently of p53. EXPLANATION OF THE INVENTION

The inventors have found a new family of compounds with the indolenin nucleus substituted by fluorine atoms that present

antitumor properties (apoptotic properties) against different cancer cell lines. Therefore, these compounds are useful for the treatment and / or prevention of cancer.

In particular, the inventors have found that the compounds of the present invention promote apoptosis in several tumor cell lines independently of the p53 protein. Under normal conditions, by detecting DNA damage, the p53 protein prevents the cell from replicating by stopping the cell cycle in the G1 phase, or inferred, allowing DNA repair. However, it induces apoptosis if cell damage is very extensive and repair attempts fail. Any interruption in the regulation of the p53 pathway, in its functioning or in the target genes increases the possibility of tumor formation.

The fact that the compounds of the present invention promote apoptosis of tumor cells independently of the p53 protein is of great importance, since the resistance of the tumor cells to current treatments is partially due to their dependence on the p53 pathway. . Therefore, the compounds of the present invention are advantageous because they reduce the chemoresistance problems associated with many antitumor agents and are active against some carcinogenic cell lines. Thus, one aspect of the present invention is related to providing compounds of general formula (I), or their pharmaceutically salts.

acceptable, or its stereoisomers or mixture of stereoisomers,

(i) where: Ri, R ₂ , R ₄ , R5, and R ₇ , are independently selected from the group consisting of H, Cl, C (= 0) 0 (Ci-C ₄ ) alkyl, 0 (Ci-C ₄ ) alkyl; and (Ci-C ₄ ) alkyl; R ₃ is selected from the group consisting of H, Cl,

C (= 0) 0 (Ci-C ₄ ) alkyl, 0 (Ci-C ₄ ) alkyl, (Ci-C ₄ ) alkyl, pyrrolidinyl-1-carbonyl, and morpholin-1 -carbonyl; R ₆ is selected from the group consisting of H and (Ci-C ₄ ) alkyl; and R ₈ is selected from the group consisting of H, F, Br, (Ci-C ₄ ) alkyl, and phenyl; with the proviso that the compound of formula (I) is different from the compound of formula (I) where Ri-R ₈ = H and with the proviso that the compound of formula (I) is different from the compound of formula (I) where R1 and R ₃ -R ₈ are H and R ₂ is methoxy.

The compound of formula (I) where Ri-R ₈ = H and the compound of formula (I) where R1 and R ₃ -R ₈ is H and R ₂ is methoxy are described in Riyuan Lin et al.,

Organic Letters 201 1, vol. 13, No. 17, pp. 4998-4501. However, nothing is said in this document regarding its use in the treatment of cancer.

The term "pharmaceutically acceptable salts" used herein comprises any salt formed from non-toxic acids or bases.

pharmaceutically acceptable including inorganic or organic acids or bases. There is no restriction on salts, except that if they are used for therapeutic purposes they must be pharmaceutically acceptable. All the compounds mentioned can have an asymmetry center, and therefore, exist in different enantiomeric forms. All individual optical isomers and stereoisomers of the compounds to which it is made reference here, and mixtures thereof, are considered within the scope of protection of the present invention. Thus, any compound referred to herein may represent any form of a racemic, one or more enantiomeric forms, one or more forms

atropoisoméhcas, and mixtures of them.

Preferably, the compounds of formula (I) are those in which Ri, R ₄ and R ₅ are H. In a preferred embodiment, the compounds of formula (I) are those in which R ₆ is H.

In a more preferred embodiment, the compounds of formula (I) are those in which R ₂ and R 7 are independently selected from H and Cl.

In an even more preferred embodiment, the compounds of formula (I) are those in which R ₃ is independently selected from the group consisting of H, Cl, and alkoxycarbonyl (Ci-C ₄ ). Preferably, the

alkoxycarbonyl (Ci-C ₄ ) is ethoxycarbonyl.

In another preferred embodiment, the compounds of the formula (I) according to the above definition are those in which R ₈ is H or methyl.

The most preferred compounds of formula (I) are those of Table 1:

Table 1 :

lh HHHHHHH Methyl l¡H Methoxy Methoxy HHHHH ij HHHHH Ethyl HH lk HH 4-pyrrolidin-1 - HHHHH carbonyl

ll H H 4-morpholin-1 - H H H H H carbonyl

lm H H Ethoxycarbonyl H H H Cl H

In H H Methoxycarbonyl H H H H Phenyl

The most preferred compounds of formula (I) are those selected from the following table:

Table 2:

In a particular preferred embodiment, the compound of formula (I) is that wherein R1-R2, _R4 -Rs is H and R ₃ is ethoxycarbonyl. In another particular preferred embodiment, the compound of formula (I) is that wherein R1-R2, _R4 -R6, and Rs are HR ₃ and R ₇ are chloro.

The compounds of the present invention can be prepared simply and flexibly from commercial reagents by a variety of procedures For example, they can be prepared by the procedure illustrated in Scheme I.

Scheme I

where R ₉ is

In the above scheme, R ₆ -R8 have the same meaning as mentioned above for the compound of formula (I).

In the above scheme R ₉ is a radical selected from the group consisting of: phenyl, phenyl substituted with at least one radical selected from the group consisting of Cl, (Ci-C ₄ ) -alkoxy, (Ci-C ₄ ) -alkyl , (Ci-C ₄ ) alkoxycarbonyl, 4-pyrrolidin-1-carbonyl, and 4-morphol-1-carbonyl.

Preferably, R ₉ is selected from the group consisting of phenyl, 4-chlorophenyl, 4-methoxyphenyl, 4-ethoxycarbonylphenyl, 4-ethylphenyl, 3-chlorophenyl, 3,4-dimethoxyphenyl, 4-phenyl-1-pyrrolidine carbonyl, 4 morpholin-1-carbonylphenyl.

According to the above scheme, 3,3-difluoro-3 / - / - indole of formula (I) can be obtained by the oxidative coupling of Suzuki of a compound of formula (III) with a boronic acid of formula R ₉ -B (OH) ₂ using a palladium catalyst such as Pd (OAc) ₂ , an oxidizing agent such as TEMPO, a base such as KF and an appropriate solvent such as propionic acid. Generally, the rection is carried out at room temperature (20-25 ° C). The subsequent fluorination of the indoline ring of compound (II) obtained with a fluorinating agent such as bis-(tetrafluoroborate) of 1-chloromethyl-4-fluoro-1,4-diazoniabicyclo [2.2.2] octane (Selectfluor® ), gives rise to compounds of formula (I). The fluorination is carried out in a suitable solvent such as acetonitrile, generally at room temperature (20-25 ° C).

The compounds of formula (I) where R ₈ is Br can be prepared by a process comprising direct arylation followed by a fluorination reaction. In this particular embodiment, the compound of formula (III) wherein R ₈ = H is subjected to an activation reaction CH with a compound of formula R ₈ -l, in the presence of a palladium (II) catalyst such as palladium acetate , and a base such as cesium acetate and an appropriate solvent such as N, N-dimethylacetamide (DMA). The reaction can be carried out at a temperature between ⁰ and 130 January 10 C. The subsequent fluorination of the compound indolenine of formula (II) can be carried out as mentioned above.

The preparation procedures described above can be modified to obtain enantiopide compounds as well as mixtures of stereoisomers. It is possible to prepare specific stereoisomers or specific mixtures by various procedures, including the use of stereospecific reagents or by introducing chiral centers in the compounds during their preparation process. Furthermore, it is possible to separate the stereoisomers once the compound has been prepared by standard resolution techniques well known to the person skilled in the art.

The preparation of pharmaceutically acceptable salts of

Compounds of formula (I) can be carried out by methods known in the state of the art. For example, they can be prepared from the primary compounds which contain a basic or acid reactive center, by conventional chemical methods. Generally, said salts are prepared, for example, by reacting the free acid or base forms of those compounds with the stoichiometric amount of a pharmaceutically acceptable base or acid in water or in an organic solvent or in a mixture thereof. The compounds of the present invention may be in crystalline form, both as solvent-free compounds or as solvates (for example, hydrates) and it is understood that both forms are within the scope of protection of the present invention. Solvation methods are generally known within the art.

An important feature of the compounds of the present invention is their ability to inhibit cell growth in tested tumor lines, and in particular their ability to induce cytotoxicity by promoting apoptosis. As shown in the Examples, the compounds of the present invention have antitumor properties on two cancer cell lines. Thus, another aspect of the present invention is related to the

preparation of compounds of general formula (I), or their salts

pharmaceutically acceptable or its stereoisomers or mixture of

stereoisomers, including the compound of formula (I) where Ri-R ₈ is H and the compound of formula (I) where Ri and R ₃ -R ₈ are H and R ₂ is methoxy, for use as a medicament.

Another aspect of the present invention is related to the preparation of compounds of formula (I), or their pharmaceutically acceptable salts, or their stereoisomers or mixture of stereoisomers, including the compounds of formula (I) where Ri-R ₈ is H and the compound of formula (I) where Ri and R ₃ -R ₈ are H and R ₂ is methoxy, for use in the treatment and / or prevention of cancer, since they are active against all types of cancer that have been tested.

In a particular embodiment, compounds of formula (I), or their pharmaceutically acceptable salts, or their stereoisomers or mixture of stereoisomers for use in the treatment and / or prevention of tumors with mutated p53 are provided.

Preferably, the compounds of the present invention are especially active against acute T-cell leukemia and cervical cancer. This aspect of the invention can also be formulated as the use of the compounds of formula (I) as defined above, for the preparation of a medicament for the treatment and / or prevention of cancer in a mammal, including humans. .

The invention also relates to a method of treating cancer in a mammal, including the human being, who suffers or is susceptible to cancer, in particular to the aforementioned types of cancer, said method comprising the administration to said patient of an amount

therapeutically effective of a compound of formula (I), as defined above, together with pharmaceutically acceptable excipients or carriers.

The compounds of the present invention can be used in the same manner as other known chemotherapeutic agents. They can be used alone or in combination with other suitable bioactive compounds.

Another aspect of the present invention is related to a pharmaceutical composition containing a therapeutically effective amount of the compounds of the present invention, of a compound of formula (I) where Ri-R ₈ is H or the compound of formula (I) where Ri and R ₃ -R ₈ are H and R ₂ is methoxy, together with appropriate amounts of pharmaceutically acceptable excipients or carriers. Preferably, the compound of formula (I) is as mentioned above without including the compound of formula (I) where Ri-R ₈ = H, or a compound of formula (I) where Ri and R ₃ -R ₈ are H and R ₂ is methoxy.

The term "therapeutically effective amount" as used herein, refers to the amount of a compound that, when administered, is sufficient to prevent the development of, or relieve to some degree, one or more of the symptoms of the disease a The one that goes. The particular dose of compound administered according to this invention will of course be determined by the particular conditions surrounding the case, including the compound administered, the route of administration, the particular condition being treated, and similar considerations. The term "pharmaceutically acceptable excipients or carriers" refers to pharmaceutically acceptable materials, components or vehicles. Each component must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the pharmaceutical composition. It should also be suitable for use in contact with tissues or organs of humans and animals without too much toxicity, irritation, allergic response, immunogenicity or other problems or complications in accordance with a reasonable benefit / risk ratio. The chemotherapeutic treatment derived from the present invention is a new approach to cancer therapy and has the advantage of being useful for the treatment of various types of cancer.

Throughout the description and the claims the word "comprises" and its vanes are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and drawings are provided by way of illustration, and are not intended to be limiting of the present invention. In addition, the present invention covers all possible combinations of particular and preferred embodiments indicated herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the dose response of compound l _c from 2 μΜ to 40 μΜ in the Jurkat cell line (T lymphocytes from an acute type T leukemia, with mutated p53) at 24 hours of incubation. Viability was measured by flow cytometry and expressed as the percentage of non-apoptotic cells (annexin V APC negative).

FIG. 2 shows the dose-response of compound l _c from 5 μΜ to 40 μΜ in the HeLa cell line (cervical adenocarcinoma epithelial cell line with p53 inactivated) at 48 hours of incubation. Viability was measured by flow cytometry and expressed as the percentage of non-apoptotic cells (annexin V APC negative). FIG. 3 shows the dose response of compound l _d from 2 μΜ to 40 μΜ in the Jurkat cell line (T lymphocytes from an acute type T leukemia, with mutated p53) at 24 hours of incubation. Viability was measured by flow cytometry and expressed as the percentage of non-apoptotic cells (annexin V APC negative).

FIG. 4 shows the dose response of compound l _d from 5 μΜ to 40 μΜ in the HeLa cell line (cervical adenocarcinoma epithelial cell line with p53 inactivated) at 48 hours of incubation. Viability was measured by flow cytometry and expressed as the percentage of non-apoptotic cells

(annexin V APC negative).

EXAMPLES Unless otherwise indicated, all reactions were carried out under an atmosphere of argon and dry glassware. Commercial reagents were used without further purification. Reaction temperatures were controlled by an IKA temperature modulator. Thin layer chromatography was performed on silica gel chromatofolios 60 F254 (Merck) and developed by visualization with UV light or with a solution of KMnO ₄ .

For silica gel column purification, silica gel (particle size 35-70 pm) was used. Symmetry C18 reverse phase columns of dimensions 4.6 mm χ 150 mm, 5 pm (column A) of HPLC were used. The HPLC analyzes were performed in an apparatus composed of two solvent supply pumps, an automatic injector and a variable wavelength detector and a system controller (Breeze V3.20). MALDI-TOF analyzes were performed using the ACH matrix. IR spectra were obtained with a Thermo Nicolet Nexus spectrophotometer and absorption bands are indicated in cm ^"1. Melting points were performed in a Büchi Melting Point B-540.

Example 1: Preparation of 2- (4-chlorophenyl) -1 / - / - indole

Indole (240 mg, 2.00 mmol), (2,2,6,6-tetramethyl-piperidin-1-yl) oxyl (TEMPO) (319 mg, 2.00 mmol), KF (11 mg, were dissolved in a closed tube 2.00 mmol), p-chlorophenylboronic acid (329 mg, 2.00 mmol), palladium acetate (1.3 mg, 0.05 mmol) in propionic acid (2 mL). The mixture was stirred at room temperature for 2 h. After completion of the reaction, it was diluted with a saturated solution of Na ₂ C03 (25 ml_) and extractions were made with dichloromethane (DCM) (3 x 10 ml_). The organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure. The residue was purified by flash chromatographic column (instantaneous) (S02, hexane: AcOEt, 9: 1), obtaining 79 mg of 2- (4-chlorophenyl) -1 / - / - indole as a white solid ( 37% yield). ¹ H NMR (CDCI ₃ , 400 MHz): 8.28 (s, 1 H), 7.63 (d, J = 7.8 Hz, 1 H), 7.61 - 7.57 (m, 2H), 7.44 - 7.39 (m, 3H), 7.24 - 7.18 (m, 1 H), 7.15 - 7.10 (m, 1 H), 6.81 (d, J = 1.3 Hz, 1 H) ppm. HRMS (ESI):

calculated for (M + H ⁺ ) CuH ₀ CIN: 228.0502; Found: 228.0578.

Example 2: Preparation of 2- (4-methoxyphenyl) -1 / - / - indole

The title compound was prepared analogously to Example 1 from indole (240 mg, 2.00 mmol) and p-methoxyphenylboronic acid (304 mg, 2.00 mmol) for 24 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 7: 3),

obtaining 203.7 mg of 2- (4-methoxyphenyl) -1 / - / - indole as a white solid (46% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.24 (s, 1 H), 7.59 (d, J = 8.8 Hz, 3H), 7.40 - 7.36 (m, 1 H), 7.20 - 7.08 (m, 2H) , 6.98 (d, J = 8.8 Hz, 2H), 6.71 (dd, J = 2.1, 0.7 Hz, 1 H), 3.86 (s, 3H) ppm.

Example 3: Preparation of 2- (4-ethoxycarbonylphenyl) -1 / - / - indole The title compound was prepared analogously to Example 1 from indole (240 mg, 2.00 mmol) and p-ethoxycarbonylphenylboronic acid (388 mg, 2.00 mmol) for 24 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 9: 1),

obtaining 104.6 mg of 2- (4-ethoxycarbonylphenyl) -1 / - / - indole as a white solid (27% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.12 (d, J = 5.7

Hz, 2H), 7.72 (m, 2H), 7.65 (d, J = 7.8 Hz, 1 H), 7.42 (d, J = 6.8 Hz, 1 H), 7.23 - 7.20 (m, 1 H), 7.14 ( t, J = 7.5 Hz, 1 H), 6.97 (s, 1 H), 4.41 (q, J = 7.1 Hz, 2H), 1.42 (t, J = 7.0 Hz, 3H) ppm. HRMS (ESI): calculated for (M + H ⁺ ) C17H15NO2: 266.1 103; Found: 266.1 178.

Example 4: Preparation of 6-chloro-2- (4-chlorophenyl) -1 / - / - indole The title compound was prepared analogously to Example 1 from 6-chloroindole (310 mg, 2.00 mmol) and p-chlorophenylboronic acid (388 mg, 2.00 mmol) for 24 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 9: 1),

obtaining 149.2 mg of 6-chloro-2- (4-chlorophenyl) -1 / - / - indole as a white solid (33% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.27 (s, 1 H), 7.59 - 7.55 (m, 2H), 7.54 - 7.50 (m, 1 H), 7.44 - 7.41 (m, 2H), 7.39 ( s, 1 H), 7.10 (dd, J = 8.4, 1 .8 Hz, 1 H), 6.78 (d, J = 1.5 Hz, 1 H) ppm. HRMS (ESI): calculated for (M + H ⁺ ) CuH ₉ CI ₂ N: 262.01 12; Found: 262.0214.

Example 5: Preparation of 2- (4-ethylphenyl) -1 H-indole

The title compound was prepared analogously to Example 1 from indole (200 mg, 1.67 mmol) and p-ethylphenylboronic acid (317 mg, 2.00 mmol) for 24 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 9: 1),

obtaining 58.7 mg of 2- (4-ethylphenyl) -1 / - / - indole as a white solid (16% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.31 (s, 1 H), 7.63 - 7.58 (m, 3H), 7.39 (dd, J = 8.0, 0.8 Hz, 1 H), 7.28 (d, J = 8.4 Hz, 2H), 7.20 - 7.16 (m, 1 H), 7.13 - 7.09 (m, 1 H), 6.81 - 6.77 (m, 1 H), 2.69 (q, J = 7.6 Hz, 2H), 1.28 ( t, J = 7.6 Hz, 3H) ppm. HRMS (ESI): calculated for (M + H ⁺ ) Ci ₆ Hi ₅ N: 222.1204; Found: 222.1303.

Example 6: Preparation of 2- (3-chlorophenyl) -1 / - / - indole

The title compound was prepared analogously to Example 1 from indole (200 mg, 1.71 mmol) and m-chlorophenylboronic acid (304.3 mg, 2.04 mmol) for 48 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 7: 3),

obtaining 41 mg of 2- (3-chlorophenyl) -1 / - / - indole as a white solid (11% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 7.64 (dd, J = 6.6, 4.9 Hz, 2H), 7.56 - 7.53 (m, 1 H), 7.42 - 7.39 (m, 1 H), 7.36 (d, J = 7.8 Hz, 1 H), 7.29 (ddd, J = 8.0, 1 .9, 1 .0 Hz, 1 H), 7.24 - 7.20 (m, 1 H), 7.16 - 7.1 1 (m, 1 H) , 6.85 (d, J = 1.3 Hz, 1 H) ppm.

Example 7: Preparation of 2- (2-chlorophenyl) -1 / - / - indole The title compound was prepared analogously to Example 1 from indole (200 mg, 1.65 mmol) and o-chlorophenylboronic acid (310.8 mg, 2.00 mmol) for 48 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 7: 3),

obtaining 39 mg of 2- (2-chlorophenyl) -1 / - / - indole as a white solid (10% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.78 (s, 1 H), 7.70 - 7.65 (m, 2H), 7.49 (dd, J = 7.9, 1.4 Hz, 1 H), 7.43 (dd, J = 8.1, 0.8 Hz, 1 H), 7.35 (td, J = 7.6, 1 .4 Hz, 1 H), 7.30 - 7.26 (m, 1 H), 7.25 - 7.21 (m, 1 H), 7.14 (td, J = 7.6, 1 .0 Hz, 1 H), 6.87 (dd, J = 2.1, 0.8 Hz, 1 H) ppm.

Example 8: Preparation of 2- (3,4-dimethoxyphenyl) -1 / - / - indole

The title compound was prepared analogously to Example 1 from indole (240 mg, 2.00 mmol) and 3,4-dimethoxyphenylboronic acid (364 mg, 2.00 mmol) for 24 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 6: 4),

244 mg of 2- (3,4-dimethoxyphenyl) -1 / - / - indole being obtained as a white solid (48% yield). Example 9: Preparation of 6-chloro-2- (4-ethoxycarbonylphenyl) -1 / - / - indole

The title compound was prepared analogously to Example 1 from 6-chloroindole (200 mg, 1.30 mmol) and p-ethoxycarbonylphenylboronic acid (320 mg, 1.56 mmol) for 24 h at room temperature. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 9: 1), obtaining 64.5 mg of 6-chloro-2- (4-ethoxycarbonylphenyl) -1 / - / - indole as a white solid (17% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.44 (s, 1 H), 8.14 - 8.08 (m, 2H), 7.72 - 7.68 (m, 2H), 7.54 (t, J = 7.6 Hz, 1 H) , 7.40 (d, J = 0.8 Hz, 1 H), 7.1 1 (dd, J = 8.4, 1 .8 Hz, 1 H), 6.91 (dd, J = 2.1, 0.8 Hz, 1 H), 4.41 (q , J = 7.1 Hz, 2H), 1.42 (t, J = 7.1 Hz, 3H) ppm.

Example 10: Preparation of 5-bromo-2-phenyl-1 / - / - indole

In a closed tube 5-bromoindole (200 mg, 1.02 mmol) and CsOAc (777 mg, 4.13 mmol) were added. The tube was purged 3 times with nitrogen and then iodobenzene (240 μΙ_, 2.14 mmol) and 300 μΙ_ of a solution of palladium aacetate in DMA [1 1 mg of Pd (OAc) ₂ in 1 ml of DMA] was added. The mixture was stirred under Ar atmosphere for 72 h at 125 ° C. When The reaction was complete, AcOEt was added and stirred for a few minutes.

The solution was then passed through S¡O2, washing the residue several times with AcOEt. It was then filtered and evaporated under reduced pressure. The residue was purified by flash chromatographic column (S¡O2, hexane: AcOEt 95: 5), obtaining 53 mg of 5-bromo-2-phenyl-1 / - / - indole as a white solid (19% yield ). ¹ H NMR (CDCI ₃ , 400 MHz): 7.80 (d, J = 1 .9 Hz, 1 H), 7.55 - 7.49 (m, 2H), 7.46 (dt, J = 8.2, 1 .8 Hz, 2H) , 7.42 - 7.37 (m, 2H), 7.33 (d, J = 3.3 Hz, 1 H), 7.29 (dd, J = 8.8, 1 .9 Hz, 1 H), 6.61 (dd, J = 3.2, 0.6 Hz , 1 H) ppm. HRMS (ESI): calculated for (M + H ⁺ ) C _or Hi ₀ BrN: 271 .9997;

Found: 272.007

Example 1 1: Preparation of 5-methyl-2-phenyl-1 H-indole

The title compound was prepared analogously to Example 10 from 5-methylindole (140 mg, 1.06 mmol) and iodobenzene (250 µΙ_, 2.22 mmol) for 72 h at 125 ° C. The residue was purified by flash chromatographic column (S¡O2, hexane: AcOEt, 95: 5), obtaining 53 mg of 5-methyl-2-phenyl-1 H-indole as a white solid (24% yield) . ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.23 (s, 1 H), 7.65 (d, J = 8.5 Hz, 2H), 7.42 (d, J = 8.0 Hz, 2H), 7.32 (d, J = 7.4 Hz, 1 H), 7.29 (d, J = 8.3 Hz, 2H), 7.02 (d, J = 8.2 Hz, 1 H), 6.75 (d, J = 1 .6 Hz, 1 H), 2.45 (s , 3H) ppm.

Example 12: Preparation of 7-ethyl-2-phenyl-1 H-indole The title compound was prepared analogously to Example 10 from 7-ethylindole (145 g, 1.03 mmol) and iodobenzene (160 μΙ_, 1. 40 mmol) for 72 h at 125 ° C. The residue was purified by flash chromatographic column (SOO2, hexane: AcOEt, 95: 5), obtaining 82 mg of 7-ethyl-2-phenyl-1 H-indole as a white solid (36% yield) . ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.22 (s, 1 H), 7.71 - 7.66 (m, 2H), 7.53 - 7.41 (m, 3H), 7.36 - 7.29 (m, 1 H), 7.13 - 7.01 (m, 2H), 6.84 (dd, J = 4.4, 2.0 Hz, 1 H), 2.92 (qd, J = 7.5, 2.4 Hz, 2H), 1.41 (ddd, J = 9.5, 6.7, 3.0 Hz , 3H) ppm.

Example 13: Preparation of 5-phenyl-2- (4-methoxycarbonylphenyl) -1 H-indole

The title compound was prepared analogously to Example 10 from 5-phenylindole (105 mg, 0.52 mmol) and methyl 4-iodobenzoate (199 mg, 0.72 mmol) for 72 h at 125 ° C. The residue was purified by column Flash chromatographic type (S02, hexane: AcOEt, 95: 5), obtaining 24 mg of 5-phenyl-2- (4-ethoxycarbonylphenyl) -1 / - / - indole as a white solid (14% yield) . ¹ H NMR (CDCI ₃ , 400 MHz): 8.45 (s, 1 H), 8.12 (d, J = 8.3 Hz, 2H), 7.86 (s, 1 H), 7.75 (d, J = 8.3 Hz, 2H) , 7.66 (dd, J = 8.2, 1 .0 Hz, 2H), 7.49 (d, J = 1 .5 Hz, 2H), 7.45 (t, J = 7.7 Hz, 2H), 7.33 (t, J = 7.4 Hz, 1 H), 7.00 (d, J = 2.1 Hz, 1 H), 3.95 (s, 3H) ppm.

Example 14: Preparation of 2- (4-pyrrolidine-1-carbonylphenyl) -1 H-indole The title compound was prepared analogously to Example 10 from indole (100 mg, 0.85 mmol) and 4-iodophenyl (pyrrolidin-1-yl) methanone (360 mg, 1.19 mmol) for 48 h at 125 ° C. The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 30:70), yielding 75 mg of 2- (4-pyrrolidine-1-carbonylphenyl) -1 / - / - indole as a white solid (30% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.53 (s, 1 H), 7.69 (dd, J = 8.3, 6.5 Hz, 2H), 7.61 (t, J = 8.5 Hz, 3H), 7.43 (dd, J = 1 1 .5, 6.0 Hz, 1 H), 7.24 - 7.19 (m, 1 H), 7.16 - 7.1 1 (m, 1 H), 6.87 (s, 1 H), 3.66 (s, 5H), 1.94 (s, 5H) ppm.

Example 15: Preparation of 2- (4-morpholine-1-carbonylphenyl) -1 / - / - indole

The title compound was prepared analogously to Example 10 from indole (40.8 mg, 0.34 mmol) and 4-iodophenyl (morphol-1-1) methanone (151.4 mg, 0.48 mmol) for 48 at 125 ° C The residue was purified by flash chromatographic column (S02, hexane: AcOEt, 30:70), obtaining 47 mg of 2- (4-morpholine-1-carbonylphenyl) -1 / - / - indole as a white solid (45% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 9.07 (s, 1 H), 7.67 - 7.61 (m, 3H), 7.40 (t, J = 8.8 Hz, 3H), 7.20 (d, J = 1 .0 Hz, 1 H), 7.12 (s, 1 H), 6.83 (d, J = 1 .4 Hz, 1 H), 3.72 (s, 8H) ppm. Example 16: Preparation of 4-iodophenyl (piperidin-1-yl) methanone

To a solution of 4-iodobenzoic acid (400 mg, 1.58 mmol) in

Anhydrous dichloromethane (10 ml_) and catalytic DMF (48 μΙ_) oxalyl chloride (1.6 ml_, 19 mmol) was added. The mixture was stirred at reflux temperature (65 ° C) for 12 h. After this time, the excess oxalyl chloride was distilled to obtain a yellow solid. Then, at a temperature of 0 ° C, a solution of pyrrolidine (660 μΙ, 7.9 mmol) in dichloromethane was added anhydrous (10 mL), stirring the mixture for 1 h. The mixture was then concentrated to dryness. The residue was purified by flash chromatographic column (Si0 ₂ , hexane: AcOEt, 40:60), obtaining 445 mg of 4- iodophenyl (piperidin-1-yl) methanone as a yellowish solid (94% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 7.79 - 7.71 (m, 2H), 7.28 - 7.24 (m, 2H), 3.63 (t, J = 6.9 Hz, 2H), 3.41 (t, J = 6.6 Hz , 2H), 2.01-1 .84 (m, 4H) ppm.

Example 17: Preparation of (4-iodophenyl) (morpholino) methanone

To a solution of 4-iodobenzoic acid (400 mg, 1.58 mmol) in

Anhydrous dichloromethane (10 mL) and catalytic DMF (48 µί) was added oxalyl chloride (1.6 mL, 19 mmol). The mixture was stirred at reflux temperature (65 ° C) for 12 h. After this time, the excess oxalyl chloride was distilled to obtain a yellow solid. Then at a temperature of 0 ° C a solution of morpholine (695 μί, 7.9 mmol) in anhydrous dichloromethane (10 mL) was added, stirring the mixture for 1 h. The mixture was then concentrated to dryness. The residue was purified by flash column chromatography type (S¡0 _2, hexane: AcOEt 40:60) to give 446 mg of (4- iodofen¡l) (morpholino) methanone as a yellowish solid (89%

performance). ¹ H NMR (CDCI ₃ , 400 MHz): 5 7.80 - 7.74 (m, 2H), 7.19 - 7.13 (m, 2H), 3.58 (m, 8H) ppm.

Example 18: Preparation of 3,3-difluoro-2-phenyl-3 / - / - indole

In a flask, 2-phenylindole (1.5 g, 7.37 mmol) was dissolved in anhydrous ACN (80 mL). Then Na ₂ C0 ₃ (s) (15 g) and the Selectfluor® fluorinating agent (5.74 g, 16.2 mmol) were added. The mixture was stirred at room temperature for 5 h. After completion of the reaction, diethyl ether (100 mL) was added and extractions were carried out with H ₂ 0 (5 x 30 mL) of the organic phases. The organic extracts were dried over Na ₂ S0 ₄ , filtered and evaporated under reduced pressure, obtaining 1.74 g of 3,3-difluoro-2-phenyl-3 / - / - indole as an orange solid (99% of performance). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.21 (d, J = 7.2 Hz, 2H), 7.62 - 7.46 (m, 6H), 7.31 (t, J = 7.9 Hz, 1 H) ppm. ¹⁹ F NMR

(CDCIs, 400 MHz): δ -1 16.84 (s) ppm. ¹³ C NMR (CDCI ₃ , 100 MHz): δ 169.15 (t, J = 24.8 Hz), 152.54 (t, J = 9.7 Hz), 132.26, 132.48, 129.08, 128.74 (m), 128.7 (t, J = 37.1 Hz), 128.54, 127.47, 123.07 (t, J = 256.3 Hz), 123.02, 121 .94 ppm. HRMS (ESI): calculated for (M + H ⁺ ) CuH ₉ F ₂ N: 230.0703; found:

230.0784.

Example 19: Preparation of 2- (4-chlorophenyl) -3,3-difluoro-3 / - / - indole

The title compound was prepared analogously to Example 18 from the compound of Example 1 (59 mg, 0.25 mmol) in anhydrous ACN (7 ml_), Na ₂ C0 ₃ (s) (500 mg) and the Selectfluor fluorinating agent ® (227.3 mg, 0.61 mmol). The mixture was stirred at room temperature for 2 h, obtaining 50.8 mg of 2- (4-chlorophenyl) -3,3-difluoro-3 / - / - ndol as an orange solid (77% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.13 (d, J = 8.5 Hz, 2H), 7.57 (d, J = 7.3 Hz, 1 H), 7.54 - 7.49 (m, 4H), 7.32 (td, J = 6.8, 2.1 Hz, 1 H) ppm. ¹⁹ F NMR (CDCI ₃ , 400 MHz): δ -1 17.1 1 ppm. ¹³ C NMR (CDCI ₃ , 100 MHz): δ 168.39 (t, J = 25.0 Hz), 152.63 (t, J = 9.6 Hz), 139.08 (s), 133.61 (s), 129.99 (s), 129.52 (s ), 128.96 (t, J = 24.0 Hz), 127.92 (s, J = 7.1 Hz), 127.55 (t, J = 2.8 Hz), 123.35 (s), 123.14 (t, J = 256.2 Hz), 122.29 (s ) ppm. HRMS (ESI): calculated for (M + H ⁺ ) CuH ₈ CIF ₂ N: 264.0313; Found: 264.0391.

Example 20: Preparation of ethyl 4- (3,3-difluoro-3 / - / - indol-2-yl) benzoate

The title compound was prepared analogously to Example 18 from the compound of Example 3 (70 mg, 0.264 mmol) in anhydrous ACN (7 mL), Na ₂ C0 ₃ (s) (600 mg) and the Selectfluor fluorinating agent ® (236.1 mg, 0.63 mmol). The mixture was stirred at room temperature for 3 h, obtaining 58.6 mg of ethyl 4- (3,3-difluoro-3 / - / - indole-2-yl) benzoate as an orange solid (74% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.26 (d, J = 8.3 Hz, 2H), 8.20 - 8.17 (m, 2H), 7.61 - 7.51 (m, 3H), 7.37 - 7.31 (m, 1 H ), 4.43 (q, J = 7.1 Hz, 2H), 1.43 (t, J = 7.1 Hz, 3H) ppm. ¹⁹ F NMR (CDCI ₃ , 400 MHz): -1 17.71 ppm. ¹³ C NMR (CDCI ₃ , 100 MHz): δ 168.61 (t, J = 25.0 Hz), 166.20 (d, J = 31 .9 Hz), 152.50 (t, J = 9.5 Hz), 133.80 (s), 133.63 (s), 132.76 (t, J = 2.7 Hz), 130.15 (s), 130.74 - 129.73 (m), 129.32 - 128.77 (m), 128.58 (s), 128.28 (s), 123.38 (s), 123.06 ( s), 122.59 (s), 61 .62 (s, J = 36.8 Hz), 14.52 (s) ppm. HRMS (ESI): calculated for (M + H ⁺ ) Ci ₇ Hi ₃ F ₂ N0 ₂ : 302.0914;

Found: 302.0993.

Example 21: Preparation of 6-chloro-2- (4-chlorophenyl) -3,3-difluoro-3 / - / - indole (le), The title compound was prepared analogously to Example 18 from the compound of Example 4 (80 mg, 0.34 mmol) in anhydrous ACN (7.5 ml_), Na ₂ C0 ₃ (s) (340 mg) and the Selectfluor fluorinating agent ® (282.5 mg, 0.76 mmol). The mixture was stirred at room temperature for 2 h, obtaining 71 mg of 6-chloro-2- (4-chlorophenyl) -3,3-difluoro-3 / - / - indole as an orange solid (70% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.12 (d, J = 8.6 Hz, 2H), 7.55 - 7.46 (m, 4H), 7.30 (dd, J = 7.8, 1.7 Hz, 1 H) ppm. ¹⁹ F NMR (CDCIs, 400 MHz): δ -1 16.33 ppm. ¹³ C NMR (CDCI ₃ , 100 MHz): δ 169.74 (t, J = 25.0 Hz), 153.99 (t, J = 9.7 Hz), 139.61 (s, J = 10.5 Hz), 139.50 (s), 130.18 (s ), 129.61 (s), 127.72 (s, J = 18.9 Hz), 127.24 (dd, J = 26.3, 22.6 Hz), 127.19 (s), 124.09 (s), 123.02 (s), 122.53 (t, J = 256.6 Hz) ppm. HRMS (ESI):

calculated for (M + H ⁺ ) C14H7CI2F2N: 297.9924; Found: 298.0009.

Example 22: Preparation of 2- (3-chlorophenyl) -3,3-difluoro-3 / - / - indole (Ih)

The title compound was prepared analogously to Example 18 from the compound of Example 6 (10 mg, 0.044 mmol) in anhydrous ACN (1.5 mL), Na ₂ C0 ₃ (s) (100 mg) and the agent Selectfluor® fluorine (34 mg, 0.09 mmol). The mixture was stirred at room temperature for 2 h, obtaining 7.3 mg of 2- (3-chlorophenyl) -3,3-difluoro-3 / - / - indole as an orange solid (63% yield). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.20 (s, 1 H), 8.06 (d, J = 7.7 Hz, 1 H), 7.61 - 7.52 (m, 7H), 7.47 (t, J = 7.9 Hz , 2H), 7.33 (td, J = 7.1, 1 .6 Hz, 2H) ppm. ¹⁹ F NMR (CDCI ₃ , 400 MHz): δ -1 17.49 ppm. HRMS (ESI): calculated for (M + H ⁺ ) CuH ₈ CIF ₂ N: 264.0313; Found: 264.039.

Example 23: Preparation of ethyl 4- (6-chloro-3,3-difluoro-3 / - / - indol-2-yl) benzoate

The title compound was prepared analogously to Example 18 from the compound of Example 9 (49 mg, 0.16 mmol) in anhydrous ACN (3.2 mL), Na ₂ C0 ₃ (s) (160 mg) and the Selectfluor fluorinating agent ® (134.1 mg, 0.36 mmol). The mixture was stirred at room temperature for 5 h, obtaining 49 mg of ethyl 4- (6-chloro-3,3-difluoro-3 / - / - indol-2-yl) benzoate as an orange solid (86% of performance). ¹ H NMR (CDCI ₃ , 400 MHz): δ 8.25 (d, J = 8.4 Hz, 2H), 8.20 - 8.17 (m, 2H), 7.57 (s, 1 H), 7.53 - 7.49 (m, 1 H) ,

7.33 (dd, J = 7.8, 1 .7 Hz, 1 H), 4.42 (t, J = 7.1 Hz, 2H), 1.44 (d, J = 7.1 Hz, 3H) ppm. ¹⁹ F NMR (CDCI ₃ , 400 MHz): δ -1 16.94 ppm HRMS (ESI): calculated for (M + H ⁺ ) C17H12CIF2NO2: 336.0525; Found: 336.0596.

Example 24: Biological assays for the detection of antitumor activity

An exploration of the antitumor activity of the compounds of formula (I) in Jurkat cells was carried out and a more detailed study of the compounds with better activity in Jurkat and HeLa cells was performed. Cell culture

Jurkat human cell lines (T lymphocytes from acute T-cell leukemia) and HeLa (epithelial cell line of

cervical adenocarcinoma) were obtained from the European Cell Culture Collection.

The Jurkat cell line grew in RPMI-1640 medium and the HeLa cell line in DMEM. Both culture media were supplemented with 10% inactivated fetal serum (veal), 1% glutamine, and 1% penicillin-streptomycin and were grown at 37 ° C in an atmosphere dampened with 5% carbon dioxide.

Reagents Dimethyl sulfoxide (DMSO) was obtained from Sigma Chemicals Co. (St Louis,

MO, USA). The propidium iodide ("propidium iodide", Pl) was obtained from Bender MedSystems (Vienna, Austria). Annexin V-APC was obtained from eBioscience (St Diego, USA). Analysis of apoptosis by flow cytometry

0.25-0.3x10 ⁶ cells were washed in phosphate buffered saline (PBS), resuspended in 100 μΙ of annexin binding buffer and incubated with 2-5 μΙ of Annexin V-Allophycocyanin (APC) .

After a 15 min incubation in the dark at room temperature, 100 μΙ of annexin binding buffer with 5 μΙ of propidium iodide ("propidium iodide", Pl) (20 μg / ml) was added just prior to analysis by flow cytometry. Data were analyzed with the appropriate program. Cell viability was measured by the externalization analysis of the

phosphatidylserine and the incorporation of Pl, and is expressed as the percentage of annexin-V cells and double negative Pl.

Apoptosis, or programmed cell death, is a general physiological mechanism for the removal of unwanted cells. It is characterized by chromatin condensation, a reduction in cell volume, and DNA cutting carried out by endonucleases that results in fragments of oligonucleosomal length. Apoptosis is also accompanied by a loss of asymmetry of the phospholipid membrane, resulting in the exposure of phosphatidylserine on the cell surface. The expression of phosphatidylserine on the cell surface plays an important role in the recognition and elimination of apoptotic cells carried out by macrophages. This is one of the earliest events of the apoptotic process. The method for the detection of apoptotic cells by flow cytometry uses the binding of annexin V labeled with a fluorochrome to phosphatidylserine. In addition, the plasma membrane is disturbed during late apoptosis but also during necrosis, since it becomes permeable to substances such as Pl. Pl is a fluorescent molecule with a molecular weight of 668.4 Da that is sandwiched between double nucleic acids chain and that can be used to stain the DNA. Pl is excluded by viable cells but can penetrate the cell membranes of dying and dead cells.

Therefore, viable cells are annexin-V and Pl double negative, early apoptotics are annexin-V positive and Pl negative while late apoptotic cells are annexin V and Pl double positive. These three populations are indicative of apoptosis. A fourth population of positive Pl cells correlates with necrotic cells.

Results 1. Exploratory test of cell viability in Jurkat cells An exploration of the effect of some compounds of formula (I) on the Jurkat cell line (T lymphocytes from an acute T-cell leukemia) was performed at a single maximum dose of 40 μΜ during a 24-hour incubation. The structure of the tested compounds are shown in Table 1. It has been also tested the 5 compound of formula (I) where _Ri-R8 are H (compound l _0).

Jurkat cells were chosen among other leukemic tumor cell lines because they have the mutated p53 protein. 1 o All the mentioned compounds were dissolved in the minimum amount of DMSO needed to be completely dissolved. Cell viability was measured by flow cytometry. It was verified that DMSO alone did not decrease cell viability. Thus, all the effects observed in cell viability were due to the activity of these compounds.

fifteen

The results are summarized in Table 3: "+" means low activity; "++" means good activity and "+++" means very good activity; "-" means that it is not active.

twenty

Table 3:

5

2. Study of the dose-response of compounds l _n and

A dose-response analysis of compounds l _c and I _{d was performed} on Jurkat cells or cells with mutated p53 and on HeLa cells with inactivated p53.

Cell viability was measured by flow cytometry and expressed as a percentage of non-apoptotic cells (annexin V negative) with respect to untreated cells at 24 and 48 hours of incubation. Therefore, values below 5% are indicative of apoptosis or loss of cell viability.

Jurkat cells (which are T lymphocytes from an acute type T leukemia, with mutated p53) were incubated with a dose range from 2 μΜ to 40 μΜ for each compound for 24 and 48 hours. All compounds induced or apoptosis in a dose-dependent manner. The viability was measured by

flow cytometry (cf. FIG. 1 and 3).

HeLa cells (cervical adenocarcinoma epithelial cell line with inactivated p53) were incubated with a dose range from 5 μΜ to 40 μΜ for each compound for 24 and 48 hours. All compounds induced

Apoptosis in a dose-dependent manner. Viability was measured by flow cytometry (cf. FIG. 2 and 4).

The IC5 ₀ at 24 hours was calculated for each compound using the values of 0 viability obtained by flow cytometric analysis. The

Results are expressed in Table 4 as the percentage of non-apoptotic cells (annexin V negative) with respect to untreated cells at 24 and 48 hours. 5 Table 4:

These results demonstrate that these compounds have antitumor activity. In addition, the apoptosis induced by these indolenin compounds is independent of the p53 protein, an important difference from the majority of drugs currently used in cancer therapy, which induce cell cycle arrest and apoptosis through the activation of p53.

Claims

CLAIMS:

one . Compound of formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a mixture of stereoisomers,

(l) where:

Ri, R2, R ₄ , R5, and R7, are independently selected from the group consisting of H, Cl,

O (Ci-C) alkyl; and (Ci-C) alkyl;

R ₃ is selected from the group consisting of H, Cl, C (= O) O (Ci-C ₄ ) alkyl, O (Ci-C ₄ ) alkyl, (Ci-C ₄ ) alkyl, pyrrolidinyl-1-carbonyl, and morpholin-1-carbonyl; R ₆ is selected from the group consisting of H and (Ci-C ₄ ) alkyl; Y

R ₈ is selected from the group consisting of H, F, Br, (Ci-C ₄ ) alkyl, and phenyl; with the proviso that the compound of formula (I) is different from the compound of formula (I) where Ri-R ₈ = H, and with the proviso that the compound of formula (I) is different from the compound of formula (I) where R1 and R ₃ -R ₈ are H and R ₂ is methoxy.

2. Compound of formula (I) according to claim 1, wherein Ri, R ₄ and R ₅ are H.

3. Compound according to claim 2, wherein R ₆ is H.

4. Compound according to any one of claims 1-3, wherein R ₂ and R7 are independently selected from the group consisting of H and Cl.

5. Compound according to any one of claims _1-3 , wherein R ₃ independently selects from the group consisting of H, Cl, and (C1 C ₄ ) alkoxycarbonyl.

6. Compound according to claim 5, wherein the (Ci-C ₄ ) alkoxycarbonyl is ethoxycarbonyl.

7 Compound according to any of claims 1-6, wherein R ₈ is H or methyl.

8. Compound according to claim 1, which is selected from the following table:

9. Compound according to claim 8, which is selected from the following

10. Compound according to claim 9, which is selected from: compound (I) where R ₁ -R ₂ , R ₄ -R ₈ are H and R ₃ is ethoxycarbonyl, and compound (I) wherein R ₁ -R ₂ ,

R ₄ -R ₆ and Rs are H and R ₃ and R ₇ are chlorine.

1 1. Compound of formula (I), as defined in any of the

claims 1-9, including a compound of formula (I) wherein Ri-R ₈ = H or 0 a compound of formula (I) wherein R ₁ and R ₃ -R ₈ are H and R ₂ is methoxy, for use as medicine.

12. Use of the compound of the compound of formula (I), as defined in any of claims 1-9, including a compound of formula (I) wherein Ri-R ₈ = H or a compound of formula (I) wherein R ₁ and R ₃ -R ₈ are H and R ₂ is methoxy, for the preparation of a medicament for the treatment and / or prevention of cancer in a mammal, including humans.

13. Use according to claim 12, wherein the cancer is selected from the group that consists of leukemia and cervical cancer.

14. Pharmaceutical composition comprising an effective therapeutic amount of the compound of formula (I), defined in any of claims 1-9, or a compound of formula (I) wherein Ri-R ₈ = H or a compound of formula (I ) 5 where R ₁ and R ₃ -R ₈ are H and R ₂ is methoxy, together with sufficient amounts of pharmaceutically acceptable excipients or carriers.

15. Pharmaceutical composition according to claim 14, wherein the compound of formula (I) is as defined in any of claims 1-9.