MXPA97000883A - Derivatives substituted from quinazol - Google Patents

Abstract

The present invention relates to: a compound having the formula (1) wherein: X is phenyl which is optionally substituted, R and R1 are each, independently, hydrogen, halogen, alkyl, alkoxy, hydroxyl or trifluoromethyl; R2 is hydrogen, alkyl, alkoxy, hydroxyl, trifluoromethyl; Y is a radical selected from the group consisting of (2) R3 is independently hydrogen, alkyl, carboxyl, carboalkoxy, phenyl or carboalkyl, n = 2 to 4 or a pharmaceutically acceptable salt of the same, with the proviso that each R3 of the radical and can be the same or different, which are useful as antineoplastic agents

Description

DERIVATIVES SUBSTITUTED FROM QUINAZOLINE BACKGROUND OF THE INVENTION This invention relates to certain quinazoline compounds, as well as pharmaceutically acceptable salts thereof. The compounds of the present invention inhibit the action of certain protein kinases of t: \ rosin (PTK) of the growth factor receptor, thereby inhibiting the abnormal growth of certain cell types. The compounds of this invention, therefore, are anticancer agents and are useful for the treatment of cancer in mammals. In addition, this invention relates to the manufacture of these quinazolines, their use for the treatment of the cannula and the pharmaceutical preparations containing them. Protein tyrosine kinases are a class of enzymes that catalyze the transfer of a phosphate group from ATP to a tyrosine residue located on a protein substrate. Protein tyrosine kinases clearly play a role in the normal growth of cells. Many of the growth factor receptor proteins function as tyrosine kinases and it is through this process that they carry out the signaling. The interaction of growth factors with these receptors is a necessary event in the normal regulation of cell growth. However, under certain conditions, as a result of a mutation or EF: 23946 - - overexpression, these receptors may become unregulated; the result of which is an uncontrolled proliferation of cells, which can lead to tumor growth and ultimately to the disease known as cancer (Wil s AF, Adv. Cancer Res., 60, 43 (1993) and Parsons , JT; Parsons, SJ, Important Advances in Oncology, DeVita VT Ed., JB Lippincott Co., Phila., 3 (1993)). Among the kinases of the growth factor receptor and its proto-oncogenes that have been identified, and which are targets of the compounds of this invention, are the epidermal growth factor receptor kinase (EGF-R protein kinase, protein kinase). oncogene erbB) and the product produced by the erbB-2 oncogene (also referred to as the neu oncogene or HER2). Because the phosphorylation event is a necessary signal for cell division to take place and because overexpressed or mutated kinases have been associated with cancer, an inhibitory agent of this event, an inhibitor of the protein kinase of tyrosine, will have a therapeutic value for the treatment of cancer and other diseases characterized by uncontrolled or abnormal cell growth. For example, the overexpression of the receptor kinase product of the erbB-2 oncogene has been associated with human breast and ovarian cancers (Slamon, DJ, et al., Science, 244, 707 (1989)). Science, 235, 1146 (1987)). Non-regulation of the EGF-R kinase has been associated with epidermoid tumors (Reiss, M. et al., Cancer Res., 51.6254 (1991)), breast tumors (Macias, A., et. al., Anticancer Res., 7,459 (1987)) and tumors involving other major organs (Gullick, J., Brit. Med. Bull., 47, 87 (1991)). Due to the importance of the role that unregulated receptor kinases play in the pathogenesis of cancer, many recent studies have dealt with specific inhibitors of PTK as potential anti-cancer therapeutic agents (some recent reviews: Burke, TR, Drugs Future, 17, 119 (1992) and Chang, CJ, Geahlen, RL, J. Nat. Prod., 55, 1529 (1992) The compounds of this invention are certain 4-anilinoquinazolines compounds In all this patent application the ring system of quinazoline will be numbered as indicated in the following formula: It has been noted that other 4-anilinoquinazolines, which differ both in nature and in the placement of the substituents at positions 5 to 8, compared to the compounds of this invention have inhibition activity of PTK. It is known, of the European Patent application 520.722 Al, of certain 4-anilinoquinazolinas containing in the positions 5 to 8 substituents of hydrogen, chloro trifluoromethyl or nitro. None of the compounds in the aforementioned application have the unique combination of substituents contained in the compounds of the present invention. Furthermore, it is noteworthy that although an anticancer utility is claimed for the compounds of the European Patent application mentioned above, no demonstration of an in vivo anticancer effect is provided. It is known, from the European patent application 566,226 Al of certain 4-anilinoquinazolines that optionally contain in the 5 to 8 positions a variety of substituents. None of the compounds in the aforementioned application has the unique combination of substituents contained in the compounds of the present invention. Furthermore, it is noteworthy that although an anticancer utility is claimed for the compounds of the aforementioned European patent application, no demonstration of a live iji anticancer effect is provided. The only living activity described in the European Patent application mentioned above is the inhibition of growth stimulated by TGF-alpha of the hepatocyte, in rats. It is known, from the European patent application 635,498 Al, of certain 4-anilinoquinazolines that optionally have, in the 6-position, a variety of substituents, while in the 7-position they must have a halogen. None of the compounds in the aforementioned application has the unique combination of substituents contained in the compounds of the present invention. Furthermore, it is noteworthy that, although an anticancer utility is claimed for the compounds of the aforementioned European patent application, no demonstration of an anti-cancer effect in vivo is provided. The only activity in vivo described in the aforementioned European patent application is the inhibition of growth stimulated by TGF-alpha of the hepatocyte in rats. In addition, certain quinazoline inhibitors that do not have a 4-anilino group are known. It is known, from European Patent Application 602,851 Al, of certain quinazolines that do not have an anilino group in the 4-position and that optionally contain, in the 5 to 8 positions, a variety of substituents. None of the compounds in the aforementioned application has the unique combination of substituents contained in the compounds of the present invention. Furthermore, it is noteworthy that, although an anti-cancer utility is claimed for the compounds of the European Patent application mentioned above, no demonstration of a live anti-cancer effect is provided. The only active activity described in the aforementioned European patent application is the inhibition of growth stimulated by TGF-alpha of the hepatocyte in the rat. It is known, from patent application WO 95/19774, of certain heterocycles, which are inhibitors of PTKs, having a pyrimidine ring similar to the 4-anilinoquinazoles of the present invention. This application mentioned above makes no mention of the 4-anilinoquinazolines, nor by the unique combination of substituents contained in the compounds of the present invention. Furthermore, it is notable that, although an anticancer utility is claimed for the compounds of the aforementioned application, no demonstration of a live anticancer effect is provided. It is known, from the patent application WO 95/157581, of certain quinazolines optionally containing in the 5 to 7 positions a variety of substituents. None of the compounds in the aforementioned application has the unique combination of substituents contained in the compounds of the present invention. Furthermore, it is noteworthy that, although an anti-cancer utility is claimed for the compounds of the aforementioned patent application, no demonstration of an anti-cancer effect in vivo is provided. further, of the Patent applications mentioned above a number of publications describe 4-anilinoquinazolines: Fry, D. W., et. al., Science, 265, 1093 (1994), Rewcastle G. W., et. al., J. Med. Chem., 38, 3482 (1995), and Bridges, A. J., et. al., J. Med. Chem., 39, 267, (1996). None of the compounds described in these publications have the unique combination of substituents contained in the compounds of the present invention. Furthermore, it is notable that no demonstration of a live iji anti-cancer effect is described in these reports. A PTK catalyzes the transfer of a phosphate group from an ATP molecule to a tyrosine residue located on a protein substrate. Inhibitors known up to now in the art are usually competitive, either with ATP or with the protein substrate of the kinase. Some of these inhibitors, the so-called mixed competitive inhibitors, can be competitive both with ATP and with the substrate, at the same time; all these competitive inhibitors work as reversible inhibitors. The 4-anilinoquinazolines known in the art are reversible inhibitors that are competitive with ATP (Fry, D. W., et al., Science, 265, 1093 (1994)). Because the concentration of ATP in a cell is normally very high (millimolar), compounds that are competitive with ATP may lack active activity because these compounds are unlikely to reach concentrations within the cell. , which are necessary to move the ATP from its binding site. As demonstrated, the quinazoline inhibitors of this invention have the unique ability to inhibit these PTKs in an irreversible manner and, therefore, are not competitive with ATP or with the protein substrate. The compounds of this invention can function as irreversible inhibitors by virtue of the fact that they can form covalent bonds with the amino acid residues located in the active site of the enzyme. As shown below, this results in a reinforced therapeutic utility of the compounds of this invention, when compared to the type of reversible inhibitor. In particular, it is shown that it is the unique nature and combination of substituents contained in the compounds of the present invention that leads to the irreversible binding of the inhibitor to the enzyme. These unique properties of the compounds of this invention contribute to their ability to inhibit the growth of human tumors in a cancer model in vivo.

DESCRIPTION OF THE INVENTION This invention provides a compound of the formula 1: wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl, cyano, nitro, carboxyl, carboalkoxy 2 to 7 carbon atoms, carboalkyl of 2 to 7 carbon atoms, amino and alkanoylamino of 1 to 6 carbon atoms; R and R are each, independently, hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl or trifluoromethyl; R_ is hydrogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl; And it is a radical selected from the group consisting of: R- is independently hydrogen, alkyl of 1 to 6 carbon atoms, carboxyl, carboalkoxy of 1 to 6 carbon atoms, phenyl or carboalkyl of 2 to 7 carbon atoms; n = 2 to 4; or a pharmaceutically acceptable salt thereof, with the proviso that each R, of the radical Y can be the same or different. The pharmaceutically acceptable salts are those derived from these organic and inorganic acids, such as acetic, lactic, citric, tartaric, succinic, maleic, malonic, gluconic, idrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic and similarly known acceptable acids. The alkyl portion of the alkyl, alkoxy, carboalkoxyl, carboalkyl and alkanoylamino substituents include both straight chain and branched carbon chains. The carboxyl is defined as the -CO-H radical. The carboalkoxy of 2 to 7 carbon atoms is defined as the radical -C02R ", where R" is an alkyl radical of 1 to 6 carbon atoms. Carboalkyl is defined as the radical -COR "where R" is an alkyl radical of 1 to 6 carbon atoms. When X is substituted it is preferred that it be mono-, di- or tri-substituted, with the monosubstituted being the most preferred. When a compound of this invention contains an asymmetric center, this invention covers the individual R and S enantiomers, as well as the racemate, with respect to this compound. Of the compounds of this invention, preferred members include those in which R, R, and R_ are hydrogen; and those in which R, R. and R_ are hydrogen and X is unsubstituted or monosubstituted with a halogen or alkyl of 1 to 6 carbon atoms. The preparation of the compounds of this invention, grouped by Formula 9, is described below in Flow Chart A, where R, R ,, R-, R3, X and n are defined and R. is alkyl of 1 to 6 carbon atoms (preferably isobutyl). Y 'is a radical selected from the group consisting of: where each R 'is independently hydrogen, alkyl of 1 to 6 carbon atoms, carboxyl, carboalkoxy of 1 to 6 carbon atoms, phenyl or carboalkyl of 2 to 7 carbon atoms. According to the reaction sequence delineated in Flow Chart A, a 5-nitro-anthranilonitrile, of Formula 2, is heated to about 100 ° C, with or without solvent, containing an excess of dimethylformamide dimethyl acetal to obtain an amidine of Formula 3. Heating a solution of amidine 3 and aniline 4 in acetic acid, for 1 to 5 hours, gives the 6-nitro-4-anilinoquinazolines of Formula 5. The reduction of the nitro group, in the compound 5, using a reducing agent, such as iron, in an acetic acid-alcohol mixture at elevated temperature, gives the 6-amino-4-anilinoquinazolines of Formula 6. The acylation of compound 6, either with an acid chloride of Formula 7 or a mixed anhydride of Formula 8 (which is prepared from the corresponding carboxylic acid), in an inert solvent, such as tetrahydrofuran (THF), in the presence of an organic base, such as pyridine or triethylamine, gives the compounds of this invention, represented by Formula 9. In cases where compounds 7 or 8 have an asymmetric carbon atom, they can be used as the racemic mixture or as the individual R or S enantiomers, in which case the compounds of this invention will be found in the racemic or optically active forms R and S, respectively. The 5-nitro-anthranilonitriles of the Formula 2 required to prepare the compounds of this invention are either known to the artisan or can be prepared by methods known in the art, as detailed in the following references; Baudet, Recl. Trav. Chim. Holland, 43, 710 (1924); Hartmans, Recl. Trav. Chim. The Netherlands, 65, 468, 469 (1946); Taylor et al., J. Amer. Chem. Soc., 82, 6058, 6063 (1960); Taylor et al., J. Amer. Chem. Soc, 82, 3152, 3154 (1960); Deshpande; Seshadri, Indian J. Chem., 11, 538 (1973); Katrizky, Alan R.; Laurenzo, Kathlenn S., J. Org. Chem., 51 (1986); Nielas, Hans-Joachim; Bohle, Matthias; Rick, Jens-Detlev; Zeuner, Frank; Zoelch, Lothar, Z. Chem., 25 (4), 137-138 (1985).

FLOW DIAGRAM A The preparation of the compounds of this invention, grouped by Formula 12, is described later in Flow Chart B where R, R ,, R ~, X and n are described above. Each Re is independently hydrogen, phenyl or alkyl of 1 to 6 carbon atoms. According to the reaction outlined in Flow Chart B, the 6-amino-4-anilinoquinazolines of Formula 10 (prepared as indicated in Flow Chart A), are added with a cyclic anhydride, of Formula 11, in an inert solvent, such as tetrahydrofuran, in the presence of a basic catalyst, such as pyridine or triethylamine.

FLOW DIAGRAM B The representative compounds of this invention were evaluated in several standard pharmacological test procedures, showing that the compounds of this invention possess an important activity as inhibitors of tyrosine protein kinases and are antiproliferative agents. Based on the activity shown in standard pharmacological test procedures, the compounds of this invention, therefore, are useful as antineoplastic agents. The test procedures used and the results obtained are shown below. The preparation of the compounds of this invention, grouped by Formula 19, is described below in Flow Chart C, where Y ', R. and X are described above. According to the reactions outlined in Flow Chart C, 4-chloro-6-nitroquinazoline, 13 (Morley, JS and Simpson, J. Chem., Soc., 360 (1948)) is reduced to 6- amino-4-chloroquinazoline, 14, using a reducing agent, such as sodium hydrosulfite, in a two-phase system, consisting of tetrahydrofuran and water, in the presence of a small amount of a phase transfer catalyst. The acylation of compound 14, either with an acid chloride of Formula 15 or with a mixed anhydride of Formula 16 (which is prepared from the corresponding carboxylic acid), in an inert solvent, such as tetrahydrofuran (THF), in The presence of an organic base, such as pyridine or N-methyl morpholine, gives the compounds of Formula 17. In those cases where compounds 15 or 16 have an asymmetric carbon atom, they can be used as the racemic mixture or as the enantiomers individual R or S, in which case the compounds of this invention will be found in the racemic or optically active forms R and S, respectively. Heating a compound of Formula 17 with an aniline of Formula 18 in an inert solvent, such as isopropanol, gives the compounds of this invention, represented by Formula 19.

FLOW DIAGRAM C 13 14 17 The preparation of the compounds of this invention, grouped by Formula 26, is described later in Flow Chart D, where Y ', R 4 and are described above. According to the reactions outlined in Flow diagram D, the nitro group of compound 20 (prepared as indicated in Flow Chart A) is reduced to the corresponding amino compound 21, using a palladium catalyst and a hydrogen source, which can be hydrogen itself or cyclohexene. The acylation of compound 21, either with an acid chloride of Formula 22 or a mixed anhydride of Formula 23 (which is prepared from the corresponding carboxylic acid), in an inert solvent, such as tetrahydrofuran (THF), in The presence of an organic base, such as pyridine or N-methyl morpholine, gives the compounds of Formula 24. In those cases where compounds 22 or 23 have an asymmetric carbon atom, they can be used as the racemate or the individual enantiomers R or 'S, in which case the compounds of this invention will be found in the racemic-active optically active forms or R and S, respectively. Heating a compound of Formula 24 with an aniline of Formula 25, in an inert solvent, such as acetic acid, gives the compounds of this invention, represented by Formula 26.

FLOW DIAGRAM D v 20 21 22 23 'N (CH3) 2 I THF, pyridine or (C2H5) 3N rc 24 Inhibition of the Empidimer Growth Factor Receptor Kinase (EGF-R) The test compounds were evaluated for their ability to inhibit the phosphorylation of the tyrosine residue of a peptide substrate, catalyzed by the epidermal growth factor receptor kinase enzyme. . The peptide substrate (RR-SRC) has the sequence arg-arg-leu-ile-glu-asp-ala-glu-tir-ala-ala-arg-gli. The enzyme was obtained as a membrane extract from A431 cells (American Type Culture Collection, Rockville, MD). The A431 cells grew in T175 flasks to an 80% confluence. The cells were washed twice with buffered saline 2+ with phosphates (PBS), without Ca. The flasks were rotated for 1.5 hours in 20 mL of PBS with 1.0 mM ethylenediaminetetraacetic acid (EDTA) at room temperature and centrifuged at 600 g for 10 minutes. The cells were solubilized in 5x10 cells per 1 mL of cold lysis buffer (10 mM 4- (2-hydroxyethyl) -l-piperazine-ethanesulfonic acid (HEPES), pH 7.6, 10 mM NaCl, 2 mM EDTA, phenylmethylsulfonyl fluoride 1 mM (PMSF), aprotinin 10 mg / mL, leupeptin 10 mg / mL, 0.1 mM sodium orthovanadate) in a Dounce homogenizer with 10 strokes, on ice. The one used was centrifuged at 600 g for 10 minutes, first to clarify the cell debris, and the supernatant was further centrifuged at 100,000 g, for 30 minutes at 4 C. The membrane button was suspended in 1.5 mL of HNG buffer (HEPES 50 mM, pH 7.6, 125 mM NaCl, 10% glycerol). The membrane extract was aliquoted, frozen immediately in liquid nitrogen, and 3β stored at -70 C. The test compounds were prepared in stock solutions of 10 mg / mL in 100% dimethylsulfoxide (DMSO). Before the experiment, the stock solutions were diluted to 500 mM with buffer (30 mM Hepes, pH 7.4) and then diluted serially to the desired concentration. An aliquot of the A431 membrane extract (10 mg / mL) was diluted in 30 mM HEPES (pH 7.4) to give a protein concentration of 50 ug / mL. To 4 microliters of the enzyme preparation was added EGF (1 microliter at 12 ug / mL) and incubated for 10 minutes on ice, followed by 4 microliters of the test compound or buffer, this mixture was incubated on ice for 30 minutes . To this was added 33P-ATP (10 mCi / mL), diluted 1:10 in the assay buffer, together with the substrate peptide, at a concentration 0.5 mM (the control reactions had no test compound) and was allowed The reaction proceeded for 30 minutes at 30 ° C. The reaction was stopped with TCA 10% and left on ice for at least 10 minutes, after which the tubes were microfuged at maximum speed for 15 minutes. Then, a portion of the supernatants was deposited in drops on P81 phosphocellulose discs and washed twice in 1% acetic acid, then with water for 5 minutes, each followed by the scintillation count. The inhibition data for the representative compounds of the invention are shown below in TABLE 1. The IC_0 value is the concentration of the test compound needed to reduce the total amount of phosphorylated substrate by 50%. The% inhibition of the test compound was determined for at least three different concentrations and the IC-.Q value was evaluated from the dose-response curve. The% inhibition was evaluated with the following formula: % inhibition = 100 - (CPM (medication) / CPM (control)) x 100 where CPM (drug) is in units of counts per minute and is a number that expresses the amount of radioactively labeled ATP (g-33P) incorporated in the RR-SRC peptide substrate by the enzyme after 30 minutes at 30 ° C, in the presence of the test compound, as measured by liquid scintillation. CPM (control) is found in units of counts per minute and is a number that expresses the amount of radioactively labeled ATP (g-33P) incorporated in the RR-SRC peptide substrate by the enzyme after 30 minutes at 30 C, in absence of the test compound, as measured by the liquid scintillation count. The CPM values were corrected for the background accounts produced by the ATP in the absence of the enzymatic reaction. The IC5Q values reported in TABLE 1 are averages of the number of tests conducted.

TABLE 1 Inhibition of the Epidermal Growth Factor Receptor Kinase. IC50 Compound Number (uM) Tests Example 4 0.012 5 Example 5 0.198 4 Example 6 0.5 1 Example 7 0.05 1 EEjjeemmpplloo 88 00..0044 2 Example 9 0.002 20 Example 10 0.11 2 Example 11 0.056 4 Example 13 10_ 1 EEjjeemmpplloo 1144 11 ..00 1 Determination of Covalent Union of the Test Compound with Epidermal Growth Factor Receptor Kinase An aliquot of enzyme extract A431 (prepared as described above) was diluted to 50 to 100 ug / mL with 30 mM Hepes buffer at pH 7.4, which contained EGF at a concentration of 12 ug / mL so that, under standard test conditions, approximately 2% of the reaction was performed and the final concentration of EGF was 2.4 ug / mL (as in the standard assay described previously). This mixture was incubated at least 10 minutes at 4 C before being used. This enzyme preparation was used for the following dialysis test procedures. To 60 microliters of the enzyme preparation 48 microliters of the test compound was added, dissolved in 5% dimethylsulfoxide (DMSO) (or only 48 microliters of 5% DMSO for control). The concentrations of the test compound were chosen from an amount 20 to 100 times higher than the IC5Q 'value to ideally ensure an 80% to 90% inhibition. The enzyme-inhibitor solution was incubated for 45 to 60 minutes at 4 C. For the non-dialyzed control test procedures, an aliquot of 9 microliters of the enzyme-inhibitor solution was evaluated under standard protocols, as described above. For the dialysis test procedures, a 60 microliter aliquot of the enzyme-inhibitor solution was placed in a well of a Pierce Microdialyzer System 100 system and dialyzed at 4 ° C against 30 mM Hepes, which contained 1.25 ug / mL of EGF, for 24 hours, with two changes of buffer (minimum 3 hours of dialysis before each change). Membrane slices with a molecular weight of 8000 were used. An aliquot of 9 microliters (at least in duplicate) of the dialyzed solution was evaluated for its activity by the standard protocol, as described above. The enzyme without added test compound retains 50 to 90% of its initial activity after dialysis. Solutions subjected to dialysis of the test compound without the added enzyme were also evaluated, to ensure that the compounds are dialysable. If the enzymatic activity is not recovered after dialysis, then it is determined that the test compounds are covalently bound (irreversible inhibition). If the enzyme activity is recovered to a large extent after dialysis, then it is determined that the test compound binds non-covalently (reversible inhibition). The determinations of covalent binding can be expressed as the% of Recovered Activity, which is calculated using the following formula that uses% inhibition before and after dialysis: % of Recovered Activity = ((% inhibition (pre-dialysis) -% Inhibition (ost-dialysis)) /% Inhibition (pre-dialysis)) x 100 A value of% of Recovered Activity close to 100% indicates non-covalent binding (reversible inhibition). A value for% of Recovered Activity much less than 100% indicates covalent bond (irreversible inhibition). The results obtained for the determinations of the covalent binding to the EGF-R kinase, for the representative compounds of this invention are given below in TABLE 2. For purposes of comparison, the TABLE 2 also provides connection data for N- (3-bromophenyl) -6,7-dimethoxy-4-quinazolinamine. This quinazoline inhibitor has been identified as a potent inhibitor of the EGF-R kinase (Fry, D. W., et al., Science, 265, 1093 (1994); Rewcastle G. W., et. al., J. Med. Chem., 38, 3482 (1995), and Bridges, E. J., et. al., J. Med. Chem., 39, 267, (1996)) and is included in European Patent Application 566,226 Al. The results of the independent multiple evaluations for each compound evaluated are given in TABLE 2: TABLE 2 Determination of Covalent Link to Receptor Kinase Epidermal Growth Factor Compound% Recovery Activity Determination Example 4 20 11 17 Covalent (link (irreversible) Example 9 9 Covalent (irreversible link) N- (3-bromophenyl) 102 70 107 Non-Covalent -6,7-dimethoxy-4- (reversiquinazolinamine bond ble) The results of TABLE 2 show that the compounds of this invention inhibit the EGF-R kinase irreversibly by forming a covalent bond with an amino acid residue located on the enzyme. In this regard, they are different from the usual 4-anilinoquinazolines, such as N- (3-bromophenyl) -6,7-dimethoxy-4-quinazolinamine, which binds in a reversible manner.

As will be delineated below, this difference in the binding capabilities between the compounds of this invention and the quinazolines and the usual quinazoline inhibitors of the prior art leads to a significantly improved biological activity and, therefore, to greater utility. therapy.

Inhibition of Cell Growth As Was Measured by the 3 Incorporation of (H) -thymidine (Tritiated Thymidine) The representative compounds of this invention were evaluated for their ability to inhibit the growth of the cell lines described below, in vitro. Inhibition is quantified by measuring the decrease in the incorporation of radiolabeled thymidine when the cells grow in the presence of the inhibitor. The A431 and SKBR3 cell lines were obtained from American Type Culture Collection, Rockville, MD. Neu-3T3 cells were obtained by transfecting the NIH 3T3 mouse fibroblasts with an activated oncogene, Neu rat. NHEK cells are obtained from Clonetics (San Diego, California). The cells grew routinely in a humidified incubator, in air with 5% CO ". These cell lines depend on the growth factors, which are ligands for receptor tyrosine kinases, which are the targets of the compounds of this invention and have the following characteristics: A431: Human squamous cell carcinoma cells, which overexpress EGFR . Neu-3T3: NIH 3T3 cells transfected with activated Neu oncogene. NHEK: Normal human epidermal keratinocytes, dependent on EGF. SKBR3: Human breast cancer cells that overexpress the ErbB2 gene.

The cell lines were cultured in appropriate media, as described below: A431: Modified media Dulbecco's Eagles, high in glucose, BRL / Gibco (10% Fetal Bovine Serum (FBS), Glutamine, Penicillin-Streptomycin) Dulbecco, R., Freeman, G. Virology 8, 396 (1959). Neu-3T3: Dulbecco Eagles modified media, with high glucose concentration (10% Fetal Bovine Serum, Glutamine, Penicillin-Streptomycin). SKBR3: Roswell Park Memorial Institute 1640 W / GLU (10% FBS, GLU, PS) Moore, G.E., Gerner, R.E. and Franklin, H.A. A.M.A., 199, 516 (1967). NHEK: Keratinocyte Growth Medium, Clonetics Boyce, S.T. and Ham, R.G. In Vitro 17, 239 (Abstract No. 159) (1981). The cells were seeded at a concentration of 10,000 cells / well, in 96-well plates, in complete media and allowed to grow to the logarithmic phase. At this stage, the complete medium was replaced with medium containing 0.5% FBS (for cells growing in 10% FBS) or media lacking epidermal growth factor (EGF) (for cells growing in free medium serum). After an overnight incubation in media with low serum content (or lacking EGF), the compounds that were evaluated were added, and the cells remained in the presence of the compounds for 48 to 72 hours. The media with the test compound were removed and complete medium was added again. The cells were allowed to grow for 18 hours. This was followed 3 by incubation in (H) thymidine (1 mCi / mL, in serum / EGF medium) for 4 hours. The cells were lysed in 0.5 M NaOH for at least 30 minutes at 37 ° C and the radioactivity was analyzed. The cell growth inhibition data is given below in TABLE 3. The IC -.Q value is the concentration of the test compound necessary to reduce the amount of (3 H) thymidine incorporation by 50%. The% inhibition of the evaluated compound was determined for at least three different concentrations and the evaluated value of C, 0 is evaluated from the dose-response curve. The% inhibition was evaluated with the following formula: % inhibition = 100 - (CPM (medication) / CPM (control) x 100 where CPM (medicine) is in units of accounts per minute and is the number that expresses the amount of 3 (H) thymidine incorporated into the DNA when the cells grow in the presence of the test compound, as measured by the liquid scintillation count. CPM (control) is found in units of counts per minute and is a number that expresses the amount of (H) thymidine incorporated into the DNA when the cells grow in the absence of the test compound, as measured by the liquid scintillation count.

TABLE 3 Inhibition of Cell Growth as Measured by the Incorporation of (H) Thymidine (IC5Q) Compound: or A431 SKBR3 NHEK NEU-3T3 (uM) (UM) (uM) (uM) Example 4 0.07 50 0.17 50 Example 5 0.825 0.30 0.17 10 Example 6 27 > 50 4.5 s or Example 7 0.45 5.5 0.45 7.5 Example 8 0.22 7 0.5 0.3 Example 9 0.011 1.057 0.002 0.002 Example 10 60 > 50 > 50 15 Example 11 0.8 4 0.85 0.4 In Vivo Inhibition of Growth of Human Epidermoid Tumors (A431) Female BALB / c nu / nu mice (Charles River, Wilmington, MA) were used in the standard iri vivo pharmacological test procedures. Human epidermoid carcinoma cells A431 (American Type Culture Collection, Rockville, Maryland # CRL-155) grew in vitro as described above. A unit of 5 x 10 cells was injected SC in the mice. When the tumors achieved a mass of between 100 and 150 mg, the mice were randomly mixed into treatment groups (day zero). The mice were treated IP once a day either on days 1, 5 and 9 or on days 1 to 10 after treating them with doses of either 80, 40 or 20 mg / Kg / dose of the compound to be evaluated, in Klucel at 0.2%. The control animals did not receive medication. The mass of tumor 2 was determined every 7 days (length x width) / 2 during the 28 days after treatment. The relative growth of the tumor (average tumor mass at day 7, 14, 21 and 28, divided by the average tumor mass at day zero) was determined for each treatment group. The% T / C (Tumor / Control) was determined by dividing the relative growth of the tumor of the treated group by the relative growth of the tumor of the placebo group and multiplying by 100. A compound is considered to be active if the% T / C is find that it is less than or equal to 42%. The inhibition results obtained for the compound of Example 9 are given below in TABLE 4.

TABLE 4 In Vivo Inhibition of Growth of Human Epidepnoid Tumors (A431) in Mice by Compound of Example 9 Dose RTG% T / CC RTG% T / CC RTGb% T / CC RTG% T / CC S / Td IP Day Day Day Day (mg / Kg / 7 16 21 28 dosia) * Control 3.68 7.91 11.41 15.04 10/10 * 80 0.71 18 0.91 11 1.07 9 1.36 9 5/5 * 40 1.48 40 2.23 28 3.05 27 4.04 27 5/5 * 20 1.72 47 2.69 34 4.33 38 6.18 41 5/5 ** 20 0.75 20 1.01 13 1.25 11 2.53 17 5/5 a) Drugs administered IP on days 1, 5, 9 * or on days 1 to 10 **. Average Tumor Mass at day 7, 14, 21, 28 b) Relative Tumor Growth = Average Tumor Mass at day 0 Relative Tumor Growth of Treated Group c)% T / C = X 100 Relative Growth of the Tumor of the Placebo Group d) S / T = No of Survivors / No. Treated to Day +28 after Tumor Treatment.

The ability of the compound of Example 9 and of N- (3-bromophenyl) -6,7-dimethoxy-4-quinazolinamine to inhibit the growth of human epidermoid tumors (A431) in vivo are compared below, in TABLE 5 The N- (3-bromophenyl) -6,7-dimethoxy-4-quinazolinamine was chosen as the comparison compound because this quinazoline inhibitor has been identified as a potent inhibitor of the EGF-R kinase (Fry, DW , et al., Science, 265, 1093 (1994), Re Castle GW, et al., J. Med. Chem., 38, 3482 (1995), Bridges, AJ, et al., Med. Chem. , 39, 267 (1996)) and because it is grouped in European Patent Application 566,226 Al.

TABLE 5 A Comparison of In Vivo Inhibition by the Compound of Example 9 and N- (3-Bromophenyl) -6,7-dimethoxy-4-quina2? Lin-amine, from the Growth of Human Epidermoid Tumors (A431) in Mice. Dosage 20 mg / Kg / dose IP. Composite - RTGb% T / CC RTGb% T / CC RTGb% T / CC RTGb% T / CC S / Td to Day D to Day Day 7 14 21 28 * Control 3.18 5.65 7.79 10.3 10/10 Example 9 1.11 35 1.26 22 1.51 19 2.55 22 14/15 NN - ((33 - bbrroo-- 33..0033 9955 66..5588 1 11166 1100..55 128 14.47 140 15/15 mofenil) - 6, 7-dimethoxy- 4-quinazo-linamine - - a) The drugs administered IP on days 1 to 15. Average Tumor Mass at day 7, 14, 21, 28 b) Relative Tumor Growth = Average Tumor Mass at day 0 Relative Growth of the Tumor of the Treaty Group c)% T / C = X 100 Relative Growth of the Tumor of the Placebo Group d) S / T = No. of Survivors / No. Treated to Day +28 after Tumor Treatment.

As shown in TABLES 4-5, the compounds of this invention inhibit the growth of human tumors in mammals and, therefore, are useful as antineoplastic agents. In this regard, they are very different from the usual 4-anilinoquinazolines, such as N- (3-bromophenyl) -6,7-dimethoxy-4-quinazolinamine, which lacks antineoplastic activity. The ability of the compound of Example 9 to inhibit the growth of human epidermoid tumors (A431) in vivo was compared with two structurally similar compounds, N- (4- ((3-methylphenyl) amino) -6-quinazolinyl) -7- fluoro-2-? ropenamide (referred to as Comparator A) and N- (4- ((3-bromophenyl) amino) -6-quinazo linyl) -butanamide (referred to as Comparator B), both of which are covered by European Patent Application 635,488A1 and 566,226 Al, respectively. The results of these comparisons are shown in TABLES 6 and 7.

TABLE 6 A Comparison of In Vivo Inhibition by the Compound of Example 9 and N- (4- ((3-methylphenyl) amino) -6-quinazolinyl) -7-fluoro-2-propenamide (Comparator A) of Growth Human Epidermoid Tumors (A431) in Mice. Compound RTG% t / cc RTG% T / CC RTG% T / CC S / Td Day Day Day 7 14 21 Control 5.52 11.63 7.79 10/10 Example 9 1.25 18 * 2.50 21 * 3.77 24 * 10/10 (80 mg / Kg) Comparator A 3.39 61 5.60 48 7.68 48 10/10 (80 mg / Kg) Example 9 0.79 14 * 1.39 12 * 2.55 16 * 10/10 (20 mg / Kg) Comparator A 4.82 87 7.36 63 8.75 56 9/10 (20 (mg / Kg) a) Medicines administered IP. The control and doses of 80 mg / kg were administered on days 1, 5 and 9; Doses of 20 mg / Kg were administered on days 1 to 10. Average Tumor Mass on day 7, 14, 21 b) Relative Tumor Growth = Average Tumor Mass at day 0 Relative Tumor Growth of the Treated Group c) % T / C = X 100 Relative Growth of the Tumor of the Placebo Group d) S / T = No. of Survivors / No. Treated to Day +21 after Tumor Treatment. * Indicates statistical significance of p less than 0.01.

TABLE 7 A Comparison of In Vivo Inhibition by the Compound of Example 9 and N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -butanamide (Comparator B) of the Growth of Human Epidermoid Tumors (A431) ) in Mice. CompuesRTG% T / CC RTG% t / cc RTG% T / CC RTG% T / CC S / Td to3 Day Day Day 7 14 21 28 Control 3.56 3.55 5.85 7.63 10/10 Ex.9 (80 * * * * mg / kg) 0.89 25 1.50 27 2.44 42 3.45 45 5/5 Comparator B (80 mg / kg) 3.37 95 5.43 98 6.21 106 10.26 142 5/5 Comparator B (20 mg / kg) 2.90 81 4.19 75 5.62 96 8.04 105 5/5 a) Drugs administered IP. The control and doses of 80 mg / kg were administered on days 1, 5 and 9; the dose of 20 mg / kg was administered on days 1 to 10. Average Tumor Mass on day 7, 14, 21, 28 b) Relative Tumor Growth = »- Average Tumor Mass at day 0 Relative Tumor Growth Treaty group c)% T / C = X 100 Relative Growth of the Tumor of the Placebo Group d) S / T = No. of survivors / No. treated at day +28 after tumor treatment. * Indicates statistical significance of p • < .0.01.

The results obtained in Tables 6 and 7 show that the compound of Example 9, a representative compound of this invention, significantly inhibited (p less than 0.01) the growth of the human epidermoid tumor alive. The structurally closest compounds of European Patent Applications 635,488 Al (Comparator A) and 566,226 Al (Comparator B) were substantially less active than the compound of Example 9 and both could not significantly reduce tumor growth at both doses evaluated. Based on the results obtained for the representative compounds of this invention, the compounds of this invention are particularly useful for the treatment, growth inhibition or eradication of neoplasms. In particular, the compounds of this invention are useful for the treatment, growth inhibition or eradication of neoplasms that express EGFR such as those of the breast, kidney, bladder, mouth, larynx, esophagus, stomach, colon. , the ovaries or the lungs. The compounds of this invention can be formulated alone or can be combined with one or more pharmaceutically acceptable carriers for administration. For example, solvents, diluents and the like and can be administered orally in forms, such as tablets, capsules, powders that can be dispersed, granules or suspensions containing, for example, from about 0.05 to 5% of the suspending agent, syrups containing, for example, from about 10 to 50% sugar and elixirs containing, for example, from about 20 to 50% ethanol and the like; or parenterally, in the form of a sterile, injectable solution or suspension containing from about 0.05 to 5% of suspending agent in an isotonic medium. Such pharmaceutical preparations may contain, for example, from about 0.05 to about 90% of the active ingredient in combination with the carrier, more usually between about 5% and 60% by weight. The effective dose of active ingredient employed may vary, depending on the particular compound employed, the mode of administration and the severity of the condition to be treated. However, in general, satisfactory results are obtained when the compounds of the invention are administered at a daily dose of from about 0.5 to about 1000 mg / Kg of animal body weight, optionally given in divided doses of two to four times a day or in the form of sustained release. For larger mammals the total daily dose is from about 1 to 1000 mg, preferably from about 2 to 500 mg. Suitable dosage forms for internal use comprise from about 0.5 to 1000 mg of the active compound in an intimate mixture with a solid or liquid pharmaceutically acceptable carrier. This dose regimen can be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be reduced proportionally, as indicated by the exigencies of the therapeutic situation.

These active compounds can be administered orally, as well as intravenously, intramuscularly or subcutaneously. Solid carriers include starch, lactose, dicalcium phosphate, icrocrystalline cellulose, sucrose and kaolin, while liquid carriers include sterile water, polyethylene glycols, nonionic surfactants and edible oils, such as corn oil, peanut oil and sesame oil, as appropriate to the nature of the active ingredient and the particular form of administration desired. Adjuvants commonly used in the preparation of pharmaceutical compositions that can be advantageously included, such as flavoring agents, coloring agents, preservatives and antioxidants, for example, vitamin E, ascorbic acid, BHT and BHA. Preferred pharmaceutical compositions, from the standpoint of ease of preparation and administration, are solid compositions, particularly tablets and solid-fill or liquid-fill capsules. Oral administration of the compounds is preferred. In some cases it may be desirable to administer the compounds directly to the airways in the form of an aerosol. These active compounds can also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds can be prepared as a free base or a pharmacologically acceptable salt in water, suitably mixed with a surfactant, such as hydroxypropylcellulose. Suspensions in glycerol, liquid polyethylene glycols and mixtures thereof in oils can also be prepared. Under ordinary conditions of storage and use, this preparation contains a condom to prevent the growth of microorganisms. Pharmaceutically acceptable forms for injectable use include aqueous, sterile solutions or dispersions, and sterile powders for extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the degree that it can be injected easily. It must be stable under the conditions of manufacture and storage and must be treated against the contaminating action of microorganisms such as bacteria and fungi. The carrier must be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof and vegetable oils. For the treatment of cancer, the compounds of this invention can be administered in combination with other anti-tumor substances, or with radiation. These other substances or radiation treatments can be administered at the same time, or at different times, than the compounds of this invention. These combined therapies can effect synergy and result in improved efficacy. For example, the compounds of this invention can be used in combination with mitotic inhibitors, such as taxol or vinblastine, alkylating agents such as cisplatin or cyclophosphamide, antimetabolites such as 5-fluorouracil or hydroxyurea, DNA intercalators such as adriamycin or bleomycin, topoisomerase inhibitors such as etoposide or camptothecin, and antiestrogen agents such as tamoxifen. The preparation of the representative examples of the compounds of this invention is described below.

EXAMPLE 1 N '- (2-Cyano-4-nitrophenyl) -N, N-dimethylformamidine A portion of 40.8 g of 5-nitro-anthranilonitrile and 40 mL of N, N-dimethylformamide dimethyl acetal were heated in a bath of steam for 2 hours. The solvents were removed under reduced pressure and the residue was taken up in methylene chloride. After passing this solution through Magnesol, the solvent was removed. After washing with ether, 50.8 g of N '- (2-cyano-4-nitrophenyl) -N, N-dimethylformamidine were obtained.

EXAMPLE 2 N- (3-bromophenyl) -6-nitro-4-quinazolinamine A solution of 23.74 mL of 3-bromo aniline and 40.5 g of N1 - (2-cyano-4-nitrophenyl) -N, N-dimethylformamidine in 100 mL of glacial acetic acid was stirred and heated in an oil bath at 148 C for 1.5 hours. After cooling, filtration of the resulting solid gives a quantitative yield of N- (3-bromophenyl) -6-nitro-4-quinazolinamine: mp = 267-270 ° C; mass spectrum (m / e): 345.

EXAMPLE 3 N- (3-bromophenyl) -4,6-quinazolindiamine A mixture of 34.5 g of N- (3-bromophenyl) -6-nitro-4-quinazolinamine and 16.8 g of iron powder in 150 mL of ethanol and 150 mL of glacial acetic acid, were heated in an oil bath at 120 C for 2 hours. After filtration of the solid, solid sodium carbonate was added to the filtrate, giving a solid, filtrate was filtered and the solid was extracted with methanol. The extracts were treated with charcoal and evaporated to a solid. After washing the solid with ether, 27.5 g of N- (3-bromophenyl) -4,6-quinazolindiamine were obtained: mass spectrum: (m / e): 315.

EXAMPLE 4 4- ((4- ((3-Bromophenyl) amino) -6-quinazolinyl) amino) -4-oxo- (Z) -2-butenóic acid A 15 mL portion of pyridine was added to 1.6 g of N - (3-bromophenyl) -4,6-quinazolindiamine and 0.6 g of maleic anhydride. After stirring overnight, the solvents were removed in a rotary evaporator. The solid was recovered in approximately 400 mL of hot ethanol and the insoluble material was filtered to give 0.33 g of 4- ((4- ((3-bromophenyl) amino) -6-quinazolinyl) amino) -4-oxo- ( Z) -2-butenóico: mass spectrum: (m / e): M + H 413, 415.

EXAMPLE 5 4- ((4- ((3-Bromophenyl) amino) -6-quinazolinyl) amino) -4-oxo- (E) -2-butenóic acid ethyl ester A solution of N- (3-bromophenyl) - 4,6-quinazolin diamine in 15 mL of pyridine was cooled in an ice bath and a solution of 1.22 g of ethyl fumaryl chloride in 10 mL of methylene chloride was added dropwise. After stirring for 1.5 hours, the reaction was allowed to reach room temperature. The solvents were removed under pressure and the residue was treated with water. The red solid was filtered and extracted into hot acetone. After filtering the insoluble material, the filtrate was concentrated to give 0.45 g of 4- ((4- ((3-bromophenyl) -amino) -6-quinazolinyl) amino) -4-oxo- (E) ethyl ester. -2-butenóico: pf = 259-263 ° C, mass spectrum: (m / e): M + H 441, 443.

EXAMPLE 6 N- (4- (3-Bromophenyl) amino) -6-quinazolinyl) -3-methyl-2-butenamide A solution of 1.58 g of N- (3-bromophenyl) -4,6-quinazolidiamine in 15 L of pyridine, cooled in an ice bath and a solution of 0.67 mL of 3, 3-dimethylacryloyl chloride in 7 mL of ether was added dropwise. After stirring and cooling for 2 hours, the solvents were removed under reduced pressure. The residue was treated with water and the resulting solid was recrystallized from methyl cellosolve to give 0.97 g of N- (4- (3-bromophenyl) amino) -6-quinazolinyl) -3-methyl-2-butenamide: pf = 300-301 C, mass spectrum (m / e): 396, 398.

EXAMPLE 7 N- (4- (bromophenyl) amino) -6-quinazolinyl) - (E) -2-butenamide A solution of 1.6 g of N- (3-bromophenyl) -4,6-quinazolindiamine in 15 mL of pyridine, it was cooled in an ice bath and a solution of 0.57 mL of trans-crotonoyl chloride in 6 mL of ether was added dropwise. After stirring and cooling for 2 hours, the solvents were removed under reduced pressure. The residue was treated with water and the resulting solid was recrystallized from n-butanol to give 0.69 g of N- (4- (bromophenyl) amino) -6-quinazolinyl) - (E) -2-butena ida: pf = 153-160 ° C, mass spectrum (m / e): M + H 383, 385.

EXAMPLE 8 N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2-methyl-2-propenamide A solution of 1.6 g of N- (3-bromophenyl) -4,6-quina-zolindiamine in 15 mL of pyridine, cooled in an ice bath and a solution of 0.59 mL of methacryloyl chloride in 6 mL of ether was added dropwise. After stirring and cooling for 2 hours, the solvents were removed under reduced pressure. The residue was treated with water and the resulting solid was taken up in n-butanol (heating). The addition of ether to the cooled solution gives 0.44 g of N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2-methyl-2-propenamide: mp = 40-245 ° C, mass spectrum (m / e): M + H 383, 385.

EXAMPLE 9 N- (4- ((3-Bromophenyl) amino) -6-quinazolinyl) -2-butinamide A solution of 0.50 g of 2-butynoic acid in 10 mL of tetrahydrofuran was cooled in an ice bath. A 0.79 mL portion of isobutyl chloroformate was added per 0.66 mL portion of N-methyl morphonyl. After about 1 minute, a solution of 1.6 g of N- (3-bromophenyl) -4,6-quinazolindiamine in 10 mL of pyridine was added. The reaction was allowed to reach room temperature and was stirred overnight. The solvents were removed under reduced pressure and the solid was recrystallized from n-butanol to give 1.07 g of N- (4- ((3-bromophenyl) -amino) -6-quinazolinyl) -2-butinamide: mass spectrum (m / e): 381, 383.

EXAMPLE 10 4- ((4- ((3-Bromophenyl) amino) -6-quinazolini1) amino) -4-oxo- (E) -2-butenóic acid A portion of 2.5 mL of 10 N aqueous sodium hydroxide, added to 2.3 g of the ethyl ester of 4 - ((4- ((3-bromophenyl) amino) -6-quinazolini1) amino) -4-oxo- (E) -2-butenóico (Example 5) in 25 mL of ethanol. After stirring for one hour, 2.1 mL of concentrated hydrochloric acid was added, and the reaction was stirred two more hours. The resulting solid was recrystallized from n-butanol to give 0.97 g of 4- ((4- ((3-bromophenyl) amino) -6-quinazolinyl) -amino) -4-oxo- (E) -2- butenóico: mass spectrum (m / e): M + H 413.

EXAMPLE 11 N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2,4-hexadienamide A solution of 0.67 g of 2,4-hexadienoic acid in mL of tetrahydrofuran, cooled in an ice bath. A 0.79 mL portion of isobutyl chloroformate was added followed by a 0.66 mL portion of N-methyl morpholine. After about 1 minute, a solution of 1.6 g of N- (3-bromophenyl) -4,6-quinazoline-diamine in 10 mL of pyridine was added. The reaction was allowed to reach room temperature and was stirred overnight. The solvents were removed under reduced pressure and the solid was recrystallized to give 1.0 g of N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2,4-hexadienamide: mp: 258-260 ° C.

EXAMPLE 12 N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2- cyclopentenamide A solution of 0.43 g of 2-cyclopentenoic acid in 5 mL of tetrahydrofuran was cooled in an ice bath. A 0.49 mL portion of isobutyl chloroformate was added followed by a 0.41 mL portion of N-methyl morpholine. After about 1 minute, a solution of 1.0 g of N- (3-bromophenyl) -4,6-quinazoline-diamine in 10 mL of pyridine was added. The reaction was allowed to reach room temperature and was stirred overnight. Another 0.5 equivalents of mixed anhydride was added. The mixture was stirred for 5 hours. The solvents were removed under reduced pressure and the solid was purified by chromatography on silica gel to give 0.30 g of N- (4- ((3-bromophenyl) amino) -6-quinazolini1) -2-cyclopenten-amide: mass (m / e): 409 (M + H, El).

- - EXAMPLE 13 N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2-propenamide A solution of 2.0 g of N- (3-bromophenyl) -4,6-quina zolindiamine in 10 mL of pyridine, it was cooled in an ice bath and a solution of 0.61 mL of acrylonitrile chloride in 30 mL of ether was added dropwise at 0 C. After stirring at room temperature for 3.5 hours, the solvents were removed under reduced pressure. The residue was purified by chromatography to give 0.2 g of N- (4- ((3-brom-or phenyl) amino) -6-quinazole? Nil) -2-propenamide: mass spectrum (m / e): M + H 369.

EXAMPLE 14 N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) - (3-phenyl-2-propinamide) A solution of 0.93 g of acid 3-phenyl-2-propionic in 10 mL of tetrahydrofuran, cooled in an ice bath. A 0.82 mL portion of isobutyl chloroformate was added followed by a 0.69 mL portion of N-methyl morpholine. After about 1 minute, a solution of 1.0 g of N- (3-bromophenyl) -4,6-quinazolindiamine in 7 mL of pyridine was added. The reaction was carried out at 0 C for 1 hour. The solvents were removed under reduced pressure and the solid was purified by chromatography on silica gel to give 0.01 g of N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) - (3-phenyl-2-propinamide) ): mass spectrum (m / e): 443.2, 445.2 (M + H, electroaerosol).

EXAMPLE 15 6-amino-4-chloroquinazoline A mixture consisting of 3.25 g of 4-chloro-6-nitroquinazoline, 10.8 g of sodium hydrosulfite, and 0.3 g of the phase transfer catalyst (C.8H._) .. NCH .. Cl "in 97 mL of tetrahydrofuran and 32 mL of water was stirred rapidly for 2 hours.The mixture was diluted with ether and the organic layer was separated.The organic solution was washed with brine and then dried over magnesium sulfate The solution was passed through a small column of silica gel.The solvent was removed at 30 ° C, under reduced pressure, to give 6-amino-4-chloroquinazoline, which is used in the next step without additional purification.

EXAMPLE 16 (4-chloro-6-quinazolinyl) -2-butinamide A solution of 1.64 g of 2-butynoic acid in 46 mL of tetrahydrofuran was cooled in an ice bath. A 2.34 mL portion of isobutyl chloroformate was added followed by a 4.13 mL portion of N-methyl morpholine. After about 10 minutes, this was emptied into a solution of 6-amino-4-chloroquinazoline in 46 mL of tetrahydrofuran. This mixture was stirred at room temperature for 2 hours. The mixture was poured into a mixture of brine and saturated sodium bicarbonate and extracted with ether. The ether solution was dried over magnesium sulfate and filtered. The solvent was removed giving the (4-chloro-6-quina-zolinyl) -2-butinamide as a color oil, which was used in the next step, without further purification.

EXAMPLE 17 N- (4- ((3-Bromophenyl) amino) -6-quinazolinyl) -2-butinamide A solution consisting of 1.76 g of (4-chloro-6-quinazolinyl) -2-butinamide and 1.23 g of 3 -Bromo Aniline, was refluxed under an inert atmosphere in 23 mL of isopropanol for 40 minutes. The mixture was cooled to room temperature and 200 mL of ether was added, giving 0.4 g of N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2-butyn-amide as the hydrochloride salt. Neutralization with a sodium bicarbonate solution, extraction with ethyl acetate, elimination of the solvent and recrystallization from 1-butanol gives the N- (4- ((3-bromophenyl) amino) -6-quinazo-linyl) -2-butinamide as the free base.

EXAMPLE 18 N '- (4-amino-2-cyanophenyl) -N, N-dimethylformamidine A solution of 6.0 g (27.5 mmol) of N' - (2-cyano-4-nitrophenyl) -N, N-dimethylformamidine, 33.9 g (41.8 L, 412.4 mmol) of cyclohexene, and 0.6 g of 10% Pd / C in 360 mL of methanol were refluxed for 4 hours. The hot mixture was filtered through celite. The solvent was removed and the residue was recrystallized from chloroform-carbon tetrachloride to give 4.9 g (95%) of the title compound as a light gray crystalline solid, mass spectrum (m / e): 188.9 (M + H, electroaerosol).

EXAMPLE 19 N- (3-cyano-4- (((dimethylamino) methylene) amino) phenyl) -2-butinamide To a solution of 2.01 g (23.9 mmol) of 2-butynoic acid and 2.9 mL (22.3 mmol) of chloroformate of isobutyl in 30 mL of tetrahydrofuran were stirred at 0 ° C under nitrogen atmosphere, as 2.42 g (2.63 mL, 22.3 mmol) of N-methyl morpholine were added, for more than 3 minutes. After stirring for 15 minutes, a solution of N '- (4-amino-2-cyanophenyl) -N, N-dimethylformamidine and 1.6 g (1.75 mL) was added., 15.9 mmol) of N-methyl morpholine in 25 mL of tetrahydrofuran, for more than 4 minutes. The mixture was stirred 30 minutes at 0 ° C and 30 minutes at room temperature. The mixture was diluted with 70 mL of ethyl acetate and emptied into a mixture of brine and saturated sodium bicarbonate. The organic layer was dried (MgSO.) and filtered through a pad of silica gel. The solvent was removed and the residue was stirred with 50 mL of ether. The suspended solid was collected to give 3.61 g (89%) of an off-white solid. Mass spectrum (m / e): 255.0 (M + H, electroaerosol).

EXAMPLE 20 N- (4- ((3-Bromophenyl) amino) -6-quinazolinyl) -2-butinamide A solution of 3.0 g (11.8 mmol) of N- (3-cyano-4 - (((dimethylamino) methylene) amino) phenyl) -2-butinamide and 2.23 g (12.98 mmol) of 3-bromoaniline in 18 mL of acetic acid, refluxed gently with stirring under nitrogen, for 1 hour 15 minutes. The mixture was cooled in an ice bath and a solid mass was formed. The solid was collected by filtration and washed with ether: acetonitrile 1: 1, to give a yellow solid which was recrystallized from ethanol, giving 2.51 g of N- (4- ((3-bromophenyl) amino) - 6-quinazolinyl) -2-butin-amide: mass spectrum (m / e): 381, 383. It is noted that, in relation to this date, the best method known to the applicant to carry out the present invention, it is the conventional one for the manufacture of the objects or substances to which it refers. Having described the invention as above, the content of the following is claimed as property.

Claims (23)

1. A compound of the formula: wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl, cyano, nitro, carboxyl, carboalkoxy 2 to 7 carbon atoms, carboalkyl of 2 to 7 carbon atoms, amino and alkanoylamino of 1 to 6 carbon atoms; R and R are each independently hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl or trifluoromethyl; R2 is hydrogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl; And it is a radical that is selected from the group consisting of: - - R- is independently hydrogen, alkyl of 1 to 6 carbon atoms, carboxyl, carboalkoxy of 1 to 6 carbon atoms, phenyl or carboalkyl of 2 to 7 carbon atoms; n = 2 to 4; or a pharmaceutically acceptable salt thereof, with the proviso that each R.- of Y may be the same or different.

2. The compound according to claim 1, characterized in that R, R, and R2 are hydrogen or a pharmaceutically acceptable salt thereof.

3. The compound according to claim 2, characterized in that X is substituted or unsubstituted with halogen or alkyl of 1 to 6 carbon atoms.

4. The compound according to claim 1, characterized in that it is N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2-butinamide or a pharmaceutically acceptable salt thereof.

5. The compound according to claim 1, characterized in that it is N- (4- ((bromophenyl) amino) -6-quinazolinyl) -2-methyl-2-propenamide or a pharmaceutically acceptable salt thereof.

6. The compound according to claim 1, characterized in that it is N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2,4-hexadienamide or a pharmaceutically acceptable salt thereof.

7. The compound according to claim 1, characterized in that it is N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) - (E) -2-butenamide or a pharmaceutically acceptable salt thereof.

8. The compound according to claim 1, characterized in that it is N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -3-methyl-2-butenamide or a pharmaceutically acceptable salt thereof.

9. The compound, according to claim 1, characterized in that it is 4 - ((4- ((3-bromophenyl) -amino) -6-quinazolinyl) amino) -4-oxo- (Z) -2-butenóic acid or a pharmaceutically acceptable salt thereof.

10. The compound according to claim 1, characterized in that it is 4 - ((4- ((3-bromophenyl) -a ino) -6-quinazolinyl) amino) -4-oxo- (E) -2-butenóic acid or a pharmaceutically acceptable salt thereof.

11. The compound according to claim 1, characterized in that it is the ethyl ester of 4 - ((4 - ((3-bromophenyl) -amino) -6-quinazolinyl) amino) -4-oxo- (E) -2 -butenoic or a pharmaceutically acceptable salt thereof.

12. The compound according to claim 1, characterized in that it is N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2-cyclopentenamide or a pharmaceutically acceptable salt thereof.

13. The compound, according to claim 1, characterized in that it is N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) -2-propenamide or a pharmaceutically acceptable salt thereof.

14. The compound according to claim 1, characterized in that it is N- (4- ((3-bromophenyl) amino) -6-quinazolinyl) - (3-phenyl-2-propinamide) or a pharmaceutically acceptable salt thereof .

15. A method for inhibiting the biological effects of a non-regulated tyrosine protein kinase in a mammal, characterized in that it comprises administering to the mammal an effective amount of a compound having the formula: wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl, cyano, nitro, carboxyl, carboalkoxy 2 to 7 carbon atoms, carboalkyl of 2 to 7 carbon atoms, amino and alkanoylamino of 1 to 6 carbon atoms; R and R. are each independently hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl or trifluoromethyl; R- is hydrogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl; And it is a radical selected from the group consisting of R ^ is independently hydrogen, alkyl of 1 to 6 carbon atoms, carboxyl, carboalkoxy of 1 to 6 carbon atoms, phenyl or carboalkyl of 2 to 7 carbon atoms; n = 2 to 4; or a pharmaceutically acceptable salt thereof, with the proviso that each R of the radical Y may be the same or different.

16. A method for the treatment, growth inhibition or eradication of neoplasms in a mammal, characterized in that it comprises administering to the mammal an effective amount of a compound having the formula: wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl, cyano, nitro, carboxyl, carboalkoxy of 2 to 7 carbon atoms, carboalkyl of 2 to 7 carbon atoms, amino and alkanoylamino of 1 to 6 carbon atoms; R and R are each, independently, hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl or trifluoromethyl; R2 is hydrogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl; And it is a radical selected from the group consisting of: R- is independently hydrogen, alkyl of 1 to 6 carbon atoms, carboxyl, carboalkoxy of 1 to 6 carbon atoms, phenyl or carboalkyl of 2 to 7 carbon atoms; - - n = 2 to 4; or a pharmaceutically acceptable salt thereof, with the proviso that each R., of the radical Y may be the same or different.

17. The method, according to claim 15, characterized in that the neoplasm expresses EGFR.

18. The method according to claim 16, characterized in that the neoplasm is selected from the group consisting of the chest, kidney, bladder, mouth, larynx, esophagus, stomach, colon, ovary and lung.

19. A pharmaceutical composition, characterized in that it comprises a compound having the formula: wherein: X is phenyl optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl, cyano, nitro, carboxyl, carboalkoxy 2 to 7 carbon atoms, carboalkyl of 2 to 7 carbon atoms, amino and alkanoylamino of 1 to 6 carbon atoms; R and R are each, independently, hydrogen, halogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl or trifluoromethyl; R- is hydrogen, alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxyl, trifluoromethyl; And it is a radical selected from the group consisting of R- is independently hydrogen, alkyl of 1 to 6 carbon atoms, carboxyl, carboalkoxy of 1 to 6 carbon atoms, phenyl or carboalkyl of 2 to 7 carbon atoms; n = 2 to 4; or a pharmaceutically acceptable salt thereof, with the proviso that each R-. of the radical Y may be the same or different or a pharmaceutically acceptable carrier. - -

20. A process to produce a compound of the formula: where R, R., R- and X are defined according to claim 1, Y 'is a radical selected from the group consisting of: wherein each R 'is, independently, hydrogen, alkyl of 1 to 6 carbon atoms, carboxyl, carboalkoxy of 1 to 6 carbon atoms, phenyl or carboalkyl of 2 to 7 carbon atoms; n is an integer from 2 to 4, wherein the process is characterized in that it comprises treating an anthranilonitrile of the formula: with dimethylformamide dimethyl acetal, with or without solvent, to give a compound of the formula: l1 heating the compound with an aniline of the formula: X-NH2 in an acidic organic solvent to give a 6-nitro-quinazoline of the formula: treat this compound with a reducing agent to give a 6-amino-quinazoline of the formula. reacting this compound with an acid chloride or mixed anhydride of the formulas: where R. is alkyl of 1 to 6 carbon atoms.

21. A process to produce a compound of the formula: where R, R ,, R ~ and X are defined according to claim 1 and (Z) indicates the configuration of the double bond, wherein the process is characterized in that it comprises reacting compounds of the formula: with a cyclic anhydride of the formula: wherein each R5 is, independently, hydrogen, phenyl or alkyl of 1 to 6 carbon atoms, in an inert solvent, in the presence of an organic base.

22. A process for producing a compound of the formula; where X is defined according to claim 1, Y * is defined according to claim 20, wherein the process is characterized in that it comprises, reducing a compound of the formula: with sodium hydrosulfite and a phase transfer catalyst, in a solvent mixture, comprising an inert organic solvent and water, to give a compound of the formula: reacting this compound with an acid chloride or mixed anhydride of the formula: where R. is alkyl of 1 to 6 carbon atoms, in the presence of an amine base, in an inert solvent, to give a compound of the formula: and heating the compound with an aniline of the formula: X-NH2 in an inert solvent.

23. A process to produce a compound of the formula: where X is defined according to claim 1, Y 'is defined according to claim 20, wherein the process is characterized in that it comprises reducing the compound of the formula: with a palladium catalyst and a hydrogen source, in an organic solvent, to give a compound of the formula: H2N N reacting this compound with an acid chloride or mixed anhydride of the formula: where R. is alkyl of 1 to 6 carbon atoms, in the presence of an amine base, in an inert solvent, to give a compound of the formulas and heating the compound with an aniline of the formula: X-NH2 in an acidic solvent. SUMMARY OF THE INVENTION This invention provides a compound having the formula: wherein: X is phenyl which is optionally substituted; R and R are each, independently, hydrogen, halogen, alkyl, alkoxy, hydroxyl or trifluoromethyl; R2 is hydrogen, alkyl, alkoxy, hydroxyl, trifluoromethyl; And it's a radical that is selected from the group that consists of - - R3 is independently hydrogen, alkyl, carboxyl, carboalkoxy, phenyl or carboalkyl; n = 2 to 4 or a pharmaceutically acceptable salt thereof, with the proviso that each R-. of the radical And may be the same or different, which are useful as antineoplastic agents.