ES2364030T3 - COMBINATION THERAPY OF ZD6474 WITH 5-FU AND / OR CPT-11. - Google Patents

Abstract

The use of 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline, also known as ZD6474: or one of its pharmaceutically acceptable salts and one of: a) 5-FU; 10 b) CPT-11; and c) 5-FU and CPT-11 in the manufacture of a medicament for the treatment of cancer in a warm-blooded animal such as a human being.

Description

The present invention relates to a combination for use in the treatment of a cancer, particularly a cancer involving a solid tumor, comprising one of: administration of ZD6474 in combination with 5-FU; the administration of ZD6474 in combination with CPT-11; and the administration of ZD6474 in combination with 5-FU and CPT-11; to a pharmaceutical composition comprising one of: ZD6474 and 5-FU; ZD6474 and CPT-11; and ZD6474 and 5-FU and CPT-11; to a combination product comprising one of: ZD6474 and 5-FU; ZD6474 and CPT-11; and ZD6474 and 5-FU and CPT-11, for use in a method of treating a human or animal body by therapy; to a kit comprising one of: ZD6474 and 5-FU; ZD6474 and CPT-11; and ZD6474 and 5-FU and CPT-11; when using one of: ZD6474 and 5-FU; ZD6474 and CPT-11; and ZD6474 and 5-FU and CPT-11, in the manufacture of a medicament for use in the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being who is optionally being treated with ionizing radiation.

Normal angiogenesis plays an important role in various processes, including embryonic development, wound healing and various components of female reproductive function. Undesirable or pathological angiogenesis has been associated with diseases that include diabetic retinopathy, psoriasis, cancer, rheumatoid arthritis, atheroma, Kaposi's sarcoma and hemangioma (Fan et al, 1995, Trends Pharmacol. Sci., 16 : 57-66; Folkman, 1995, Nature Medicine 1: 27-31). It is believed that the alteration of vascular permeability plays a role in both normal and pathological physiological processes (Cullinan-Bove et al, 1993, Endocrinology 133: 829-837; Senger et al, 1993, Cancer and Metastasis Reviews, 12 : 303-324). Several polypeptides with in vitro endothelial cell growth promoting activity have been identified, including the growth factors of acidic and basic fibroblasts (aFGF and bFGF) and vascular endothelial growth factor (VEGF). By virtue of the restricted expression of its receptors, VEGF growth factor activity, in contrast to that of FGF, is relatively specific for endothelial cells. Recent results indicate that VEGF is an important stimulant of both normal and pathological angiogenesis (Jakeman et al, 1993, Endocrinology, 133: 848-859; Kolch et al, 1995, Breast Cancer Research and Treatment, 36: 139 -155) and vascular permeability (Connolly et al, 1989, J. Biol. Chem 264: 20017-20024). Antagonism of the action of VEGF by sequestration of VEGF with antibodies may result in tumor growth inhibition (Kim et al, 1993, Nature 362: 841-844).

Receptor tyrosine kinases (RTKs) are important in the transmission of biochemical signals through the plasma membrane of cells. These transmembrane molecules typically consist of an extracellular domain of binding to the ligand connected through a segment in the plasma membrane to an intracellular domain with tyrosine kinase activity. Binding of the ligand to the receptor results in stimulation of tyrosine kinase activity associated with the receptor, which leads to phosphorylation of tyrosine residues both in the receptor and in other intracellular molecules. These changes in tyrosine phosphorylation initiate a cascade of signals that leads to a series of cellular responses. To date, at least nineteen differentiated subfamilies of RTK have been identified, defined by the homology of the amino acid sequence. One of these subfamilies currently comprises the fms-type tyrosine kinase receptor, Flt-1, the receptor that contains the kinase insertion domain, KDR (also called Flk-1) and another fms-type tyrosine kinase receptor, Flt -4. Two of these related RTKs, Flt-1 and KDR, have been shown to bind VEGF with great affinity (De Vries et al, 1992, Science 255: 989-991; Terman et al, 1992, Biochem Biophys. Res. Comm ., 1992, 187: 1579-1586). VEGF binding to these receptors expressed in heterologous cells has been associated with changes in the tyrosine phosphorylation state of cellular proteins and in calcium fluxes.

VEGF is a key stimulus for vasculogenesis and angiogenesis. This cytokine induces a rapid vascular growth phenotype by inducing endothelial cell proliferation, protease expression and migration, and the subsequent organization of cells to form a capillary tube (Keck, PJ, Hauser, SD, Krivi, G. , Sanzo, K., Warren, T., Feder, J., and Connolly, DT, Science (Washington DC), 246: 1309-1312, 1989; Lamoreaux, WJ, Fitzgerald, ME, Reiner, A., Hasty, KA, and Charles, ST, Microvasc. Res., 55: 29-42, 1998; Pepper, MS, Montesano, R., Mandroita, SJ, Orci, L. and Vassalli, JD, Enzyme Protein, 49: 138-162 , nineteen ninety six.). In addition, VEGF induces significant vascular permeability (Dvorak, HF, Detmar, M., Claffey, KP, Nagy, JA, van de Water, L., and Senger, DR, (Int. Arch. Allergy Immunol., 107: 233-235, 1995; Bates, DO, Heald, RI, Curry, FE and Williams, BJ Physiol. (Lond.), 533: 263-272, 2001), promoting the formation of an immature, hyperpermeable vascular network, which is characteristic of pathological angiogenesis.

It has been shown that KDR activation is only sufficient to promote all major phenotypic responses to VEGF, including proliferation, migration and survival of endothelial cells, and induction of vascular permeability (Meyer, M., Clauss, M ., Lepple-Wienhues, A., Waltenberger, J., Augustin, HG, Ziche, M., Lanz, C., Büttner, M., Rziha, HJ., And Dehio, C., EMBO J., 18: 363-374, 1999; Zeng, H., Sanyal, S. and Mukhopadhyay, D., J. Biol. Chem, 276: 32714-32719, 2001; Gille, H., Kowalski, J., Li, B., LeCouter, J., Moffat, B, Zioncheck, TF, Pelletier, N. and Ferrara, N., J. Biol. Chem., 276: 3222-3230, 2001).

Quinazoline derivatives that are inhibitors of the VEGF tyrosine kinase receptor are described in International Patent Application Publications No. WO 98/13354 and WO 01/32651. In WO 98/13354 and WO 01/32651, compounds having activity against the VEGF tyrosine kinase receptor are described while having some activity against the EGF tyrosine kinase receptor. The compound of the present invention, ZD6474, falls within the broad general description of WO 98/13354 and is given as an example in WO 01/32651.

In WO 01/32651 it is stated that the compounds of said invention:

"May be applied as a monotherapy or may involve, in addition to a compound of the invention, one or more additional substances and / or treatments. Such joint treatment can be achieved by simultaneous, sequential or separate administration of the individual components of the treatment ». Next, WO 01/32651 continues to describe examples of such joint treatment that include surgery, radiotherapy and many types of chemotherapeutic agents including 5-fluorouracil (5-FU) and irinotecan (CPT-11). Nowhere in WO 01/32651 is it specified that the use of any compound of the invention with other treatments will surprisingly produce beneficial effects.

Unexpectedly and surprisingly it has now been found that the particular compound ZD6474 used in combination with a particular selection of combination therapies, specifically with one of: 5-FU; CPT-11; and 5-FU and CPT-11, produces significantly better anti-angiogenic and / or vascular permeability reducing effects than any one of: ZD6474; 5-FU; CPT-11; and 5-FU and CPT-11 used alone. According to one aspect of the present invention, the ZD6474 used in combination with one of: 5-FU; CPT-11; and 5-FU and CPT-11 produces significantly better anticancer effects than any one of: ZD6474; 5-FU; CPT-11; and 5-FU and CPT-11 used alone. According to one aspect of the present invention, the ZD6474 used in combination with one of: 5FU; CPT-11; and 5-FU and CPT-11 produces significantly better effects on solid tumors than any one of: ZD6474; 5-FU; CPT-11; and 5-FU and CPT-11 used alone. According to one aspect of the present invention, the ZD6474 used in combination with one of: 5-FU; CPT-11; and 5-FU and CPT-11 produces significantly better effects on colorectal cancer than any one of: ZD6474; 5-FU; CPT-11; and 5-FU and CPT-11 used alone.

The anticancer effects of a combination of the present invention include, but are not limited to, the antitumor effects, the speed of response, the time for disease progression and the survival rate. The antitumor effects of a combination of the present invention include, but are not limited to, tumor growth inhibition, tumor growth retardation, tumor regression, tumor shrinkage, increased growth time again. of the tumor after cessation of treatment and the slowdown of disease progression. It is expected that when a combination of the present invention is administered to a warm-blooded animal such as a human being, in need of a treatment for cancer, with or without a solid tumor, said combination will produce an effect, measured, for example, for one or more of: the degree of the antitumor effect, the speed of response, the time for disease progression and the survival rate.

According to the present invention a combination is described for the production of an antiangiogenic effect and / or a reduction effect of vascular permeability in a warm-blooded animal such as a human being, which comprises administering to said animal an effective amount of 4- (4-Bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline, also known as ZD6474:

image 1

or of one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of one of:

a) 5-FU;

b) CPT-11; Y

c) 5-FU and CPT-11.

According to another aspect of the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before , after or simultaneously with an effective amount of one of: 5-FU; CPT-11; and 5-FU and CPT-11.

According to another aspect of the present invention there is provided a method for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of one of: 5-FU; CPT-11; and 5-FU and CPT-11.

According to another aspect of the present invention there is provided a combination for the treatment of colorectal cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before , after or simultaneously with an effective amount of one of: 5-FU; CPT-11; and 5-FU and CPT-11.

According to another aspect of the present invention there is provided a combination for the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being, which comprises administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before, after or simultaneously with an effective amount of one of: 5-FU; CPT11; and 5-FU and CPT-11; wherein ZD6474, 5-FU and CPT-11 can each optionally be administered together with a pharmaceutically acceptable carrier or excipient,

According to another aspect of the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before , after or simultaneously with an effective amount of one of: 5-FU; CPT-11; and 5-FU and CPT-11; wherein ZD6474, 5-FU and CPT-11 can each optionally be administered together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention there is provided a combination for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of one of: 5-FU; CPT-11; and 5-FU and CPT-11; wherein ZD6474, 5-FU and CPT-11 can each optionally be administered together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention there is provided a composition for the treatment of colorectal cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before , after or simultaneously with an effective amount of one of: 5-FU; CPT-11; and 5-FU and CPT-11; wherein ZD6474, 5-FU and CPT-11 can each optionally be administered together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the invention there is provided a pharmaceutical composition comprising ZD6474 or a pharmaceutically acceptable salt thereof, and 5-FU in association with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the invention there is provided a pharmaceutical composition comprising ZD6474 or a pharmaceutically acceptable salt thereof, and CPT-11 in association with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the invention there is provided a pharmaceutical composition comprising ZD6474 or a pharmaceutically acceptable salt thereof, and 5-FU and CPT-11 in association with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention there is provided a combination product comprising ZD6474 or a pharmaceutically acceptable salt and 5-FU, for use in a method of treating a human or animal body by therapy.

According to another aspect of the present invention there is provided a combination product comprising ZD6474 or one of its pharmaceutically acceptable salts and CPT-11, for use in a method of treating a body human or animal through therapy.

According to another aspect of the present invention there is provided a combination product comprising ZD6474 or one of its pharmaceutically acceptable salts and 5-FU and CPT-11, for use in a method of treating a human or animal body through therapy.

According to another aspect of the present invention there is provided a kit comprising ZD6474 or one of its salts

pharmaceutically acceptable, and 5-FU. According to another aspect of the present invention there is provided a kit comprising ZD6474 or one of its salts pharmaceutically acceptable, and CPT-11.

According to another aspect of the present invention there is provided a kit comprising ZD6474 or one of its salts pharmaceutically acceptable, and 5-FU and CPT-11. According to another aspect of the present invention a kit is provided comprising: a) ZD6474 or a pharmaceutically acceptable salt thereof in a first unit dosage form; b) 5-FU in a second unit dosage form; Y c) containers for containing said first and second pharmaceutical forms. According to another aspect of the present invention a kit is provided comprising: a) ZD6474 or a pharmaceutically acceptable salt thereof in a first unit dosage form; b) CPT-11 in a second unit dosage form; Y c) containers for containing said first and second pharmaceutical forms. According to another aspect of the present invention a kit is provided comprising: a) ZD6474 or a pharmaceutically acceptable salt thereof in a first unit dosage form; b) 5-FU in a second unit dosage form; c) CPT-11 in a third unit dosage form; Y d) containers for containing said first, second and third pharmaceutical forms;

According to another aspect of the present invention a kit is provided comprising: a) ZD6474 or a pharmaceutically acceptable salt thereof, together with a pharmaceutically excipient or vehicle acceptable, in a first unit dosage form;

b) 5-FU together with a pharmaceutically acceptable carrier or excipient, in a second pharmaceutical form unitary; Y c) containers for containing said first and second pharmaceutical forms.

According to another aspect of the present invention a kit is provided comprising: a) ZD6474 or a pharmaceutically acceptable salt thereof, together with a pharmaceutically excipient or vehicle acceptable, in a first unit dosage form;

b) CPT-11 together with a pharmaceutically acceptable excipient or vehicle, in a second pharmaceutical form unitary; Y c) containers for containing said first and second pharmaceutical forms.

According to another aspect of the present invention a kit is provided comprising: a) ZD6474 or a pharmaceutically acceptable salt thereof, together with a pharmaceutically excipient or vehicle acceptable, in a first unit dosage form;

b) 5-FU together with a pharmaceutically acceptable carrier or excipient, in a second pharmaceutical form unitary;

c) CPT-11 together with a pharmaceutically acceptable excipient or vehicle, in a third pharmaceutical form unitary; Y

d) containers for containing said first, second and third pharmaceutical forms; According to another aspect of the present invention, the use of ZD6474 or one of its salts is provided pharmaceutically acceptable and one of:

a) 5-FU; b) CPT-11; Y c) 5-FU and CPT-11 in the manufacture of a medicament for use in the production of an antiangiogenic effect and / or a reducing effect

of vascular permeability in a warm-blooded animal such as a human being.

According to another aspect of the present invention, the use of ZD6474 or one of its salts is provided pharmaceutically acceptable and one of: a) 5-FU; b) CPT-11; Y c) 5-FU and CPT-11 in the manufacture of a medicament for use in the production of an anticancer effect in a blood animal

Hot just like a human being.

According to another aspect of the present invention, the use of ZD6474 or one of its salts is provided pharmaceutically acceptable and one of: a) 5-FU; b) CPT-11; Y c) 5-FU and CPT-11 in the manufacture of a medicament for use in the production of an antitumor effect in a blood animal

Hot just like a human being.

According to another aspect of the present invention, the use of ZD6474 or one of its salts is provided pharmaceutically acceptable and one of: a) 5-FU; b) CPT-11; Y c) 5-FU and CPT-11 in the manufacture of a medicament for use in the production of an anticancer effect in a blood animal

hot just like a human being in which cancer is a colorectal cancer. According to another aspect of the present invention a combination therapeutic treatment is provided which comprises the administration of an effective amount of ZD6474 or a pharmaceutically active salt thereof acceptable, optionally together with a pharmaceutically acceptable carrier or excipient, and administration of an effective amount of 5-FU, optionally together with a pharmaceutically acceptable carrier or carrier, at a

warm-blooded animal such as a human being in need of such therapeutic treatment, in which the ZD6474 and 5-FU can be administered simultaneously, sequentially or separately and in any order. According to another aspect of the present invention a combination therapeutic treatment is provided which

comprises the administration of an effective amount of ZD6474 or a pharmaceutically active salt thereof acceptable, optionally together with a pharmaceutically acceptable carrier or excipient, and administration of an effective amount of CPT-11, optionally together with a pharmaceutically acceptable carrier or carrier, to a warm-blooded animal such as a human being that needs such therapeutic treatment, in which the ZD6474 and CPT-11 can be administered simultaneously, sequentially or separately and in any order.

According to another aspect of the present invention a combination therapeutic treatment is provided which comprises the administration of an effective amount of ZD6474 or a pharmaceutically active salt thereof acceptable, optionally together with a pharmaceutically acceptable carrier or excipient, and administration of an effective amount of 5-FU, optionally together with a pharmaceutically acceptable carrier or carrier, and the

administration of an effective amount of CPT-11, optionally together with a pharmaceutically acceptable carrier or excipient, to a warm-blooded animal such as a human being in need of such therapeutic treatment, in which the ZD6474, 5-FU and CPT-11 they can be administered simultaneously, sequentially or separately and in any order.

A combination treatment of the present invention as defined herein can be performed by the simultaneous, sequential or separate administration of the individual components of said treatment. A combination treatment, as defined herein, may be applied as a single therapy or may involve surgery or radiotherapy or an additional chemotherapeutic agent, in addition to a combination treatment of the invention. Surgery may comprise the stage of partial or complete resection of the tumor, before, during or after the administration of the combination treatment with ZD6474 described herein.

Other chemotherapeutic agents for optional use with a combination treatment of the present invention include those described in WO 01/32651.

Such chemotherapy can cover five main categories of therapeutic agents as indicated in WO 01/32651:

(i)

 other antiangiogenic agents, including agents for vascular targets;

(ii)

 cytostatic agents;

(iii) biological response modifiers (for example interferon);

(iv)

 antibodies (eg edrecolomab); Y

(v)

 antiproliferative / antineoplastic drugs and their combinations, such as those used in medical oncology; and other categories of agents are:

(saw)

 antisense therapies;

(vii) methods of gene therapy; Y

(ix) immunotherapy methods.

The administration of a multiple combination of ZD6474, 5-FU and ionizing radiation or ZD6474, CPT-11 and ionizing radiation or ZD6474, 5-FU, CPT-11 and ionizing radiation can produce effects, such as antitumor effects, greater than those achieved with any of ZD6474, 5-FU, CPT-11 and ionizing radiation used alone. The administration of a multiple combination of ZD6474, 5-FU and ionizing radiation or ZD6474, CPT-11 and ionizing radiation or ZD6474, 5-FU, CPT-11 and ionizing radiation can produce effects, such as antitumor effects, greater than those achieved with the combination of ZD6474 and 5-FU, greater than those achieved with the combination of ZD6474 and CPT-11 and greater than those achieved with the combination of ZD6474, 5-FU and CPT-11. The administration of a multiple combination of ZD6474, 5-FU and ionizing radiation or ZD6474, CPT-11 and ionizing radiation or ZD6474, 5-FU, CPT-11 and ionizing radiation can produce effects, such as antitumor effects, greater than those achieved with the combination of ZD6474 and ionizing radiation, greater than those achieved with the combination of 5-FU and ionizing radiation, greater than those achieved with the combination of CPT-11 and ionizing radiation, and greater than those achieved with the combination of 5- FU, CPT-11 and ionizing radiation.

According to the present invention a combination is described for the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of 5-FU and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention a combination is described for the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention a combination is described for the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of 5-FU, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474

or of one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of 5-FU and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474

or of one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474

or of one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of 5-FU, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention a combination is provided for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof. , before, after or simultaneously with an effective amount of 5-FU and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention a combination is provided for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof. , before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation.

According to the present invention a combination is provided for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof. , before, after or simultaneously with an effective amount of 5-FU, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation.

In particular, the cancer that involves a solid tumor is colorectal cancer.

According to another aspect of the present invention a combination is described for the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being, which comprises administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of 5-FU and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474 and 5-FU can each be administered optionally together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention a composition is described for the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being, which comprises administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474 and CPT-11 can each be administered optionally together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention a composition is described for the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being, which comprises administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before, after or simultaneously with an effective amount of 5-FU, before, after

or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474, 5-FU and CPT-11 can each optionally be administered together with a pharmaceutically acceptable excipient or vehicle .

According to another aspect of the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before , after or simultaneously with an effective amount of 5-FU and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474 and 5-FU can each optionally be administered together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before , after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474 and CPT-11 can each optionally be administered together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention there is provided a combination for the treatment of a cancer in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, before , after or simultaneously with an effective amount of 5-FU, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474, 5-FU and CPT -11 can each optionally be administered together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention there is provided a combination for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of 5-FU and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474 and 5-FU can each optionally be administered together with an excipient or pharmaceutically acceptable vehicle.

According to another aspect of the present invention there is provided a combination for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474 and CPT-11 can each optionally be administered together with an excipient or pharmaceutically acceptable vehicle.

According to another aspect of the present invention there is provided a combination for the treatment of a cancer involving a solid tumor in a warm-blooded animal such as a human being, comprising administering to said animal an effective amount of ZD6474 or one of its pharmaceutically acceptable salts, before, after or simultaneously with an effective amount of 5-FU, before, after or simultaneously with an effective amount of CPT-11 and before, after or simultaneously with an effective amount of ionizing radiation, in which ZD6474, 5FU and CPT-11 can each be optionally administered together with a pharmaceutically acceptable carrier or excipient.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt and 5-FU is provided in the manufacture of a medicament for use in the production of an antiangiogenic effect and / or a vascular permeability reducing effect in a warm-blooded animal such as a human being who is being treated with ionizing radiation.

According to another aspect of the present invention, the use of ZD6474 or one of its pharmaceutically acceptable salts and CPT-11 is provided in the manufacture of a medicament for use in the production of an antiangiogenic effect and / or a vascular permeability reducing effect in a warm-blooded animal such as a human being who is being treated with ionizing radiation.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt thereof and 5-FU and CPT-11 is provided in the manufacture of a medicament for use in the production of an antiangiogenic effect and / or a reducing effect of vascular permeability in a warm-blooded animal such as a human being who is being treated with ionizing radiation.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt and 5-FU in the manufacture of a medicament for use in the production of an anticancer effect in a warm-blooded animal such as a human being is provided that is being treated with ionizing radiation.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt and CPT-11 is provided in the manufacture of a medicament for use in the production of an anti-cancer effect in a warm-blooded animal such as a human being. that is being treated with ionizing radiation.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt thereof and 5-FU and CPT-11 is provided in the manufacture of a medicament for use in the production of an anti-cancer effect in such a warm-blooded animal. as a human being who is being treated with ionizing radiation.

In particular, the warm-blooded animal such as a human being has colorectal cancer.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt and 5-FU is provided in the manufacture of a medicament for use in the production of an antitumor effect in a warm-blooded animal such as a human being. that is being treated with ionizing radiation.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt and CPT-11 is provided in the manufacture of a medicament for use in the production of an antitumor effect in a warm-blooded animal such as a human being. that is being treated with ionizing radiation.

According to another aspect of the present invention, the use of ZD6474 or a pharmaceutically acceptable salt thereof and 5-FU and CPT-11 is provided in the manufacture of a medicament for use in the production of an antitumor effect in such a warm-blooded animal. as a human being who is being treated with ionizing radiation.

According to another aspect of the present invention, a combination therapeutic treatment is provided comprising the administration of an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable carrier or carrier, and the administration of an effective amount. 5-FU, optionally together with a pharmaceutically acceptable excipient or vehicle and the administration of an effective amount of ionizing radiation, to a warm-blooded animal such as a human being in need of such therapeutic treatment, in which the ZD6474, 5- FU and ionizing radiation can be administered simultaneously, sequentially or separately and in any order.

According to another aspect of the present invention, a combination therapeutic treatment is provided comprising the administration of an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable carrier or carrier, and the administration of an effective amount. of CPT-11, optionally together with a pharmaceutically acceptable excipient or vehicle and the administration of an effective amount of ionizing radiation, to a warm-blooded animal such as a human being in need of such therapeutic treatment, in which the ZD6474, CPT- 11 and ionizing radiation can be administered simultaneously, sequentially or separately and in any order.

According to another aspect of the present invention, a combination therapeutic treatment is provided comprising the administration of an effective amount of ZD6474 or a pharmaceutically acceptable salt thereof, optionally together with a pharmaceutically acceptable carrier or carrier, and the administration of an effective amount. 5-FU, optionally together with a pharmaceutically acceptable excipient or vehicle, and the administration of an effective amount of CPT-11, optionally together with a pharmaceutically acceptable excipient or vehicle, and the administration of an effective amount of ionizing radiation, to a Warm-blooded animal such as a human being in need of such therapeutic treatment, in which the ZD6474, 5-FU, CPT11 and ionizing radiation can be administered simultaneously, sequentially or separately and in any order.

A warm-blooded animal such as a human being who is being treated with ionizing radiation means a warm-blooded animal such as a human being who is treated with ionizing radiation before, after or at the same time as the administration of a medication or a combination treatment comprising ZD6474 and one of: 5-FU; CPT-11; and 5-FU and CPT-11. For example, said ionizing radiation can be given to said warm-blooded animal such as a human being within the period of one week before to one week after the administration of a medicament or a combination treatment comprising ZD6474 and one of: 5-FU; CPT-11; and 5-FU and CPT-11. This means that ZD6474, 5-FU, CPT-11 and ionizing radiation can be administered separately or sequentially in any order, or they can be administered simultaneously. The warm-blooded animal can experience the effect of each of ZD6474, 5-FU, CPT-11 and radiation simultaneously.

According to one aspect of the present invention the ionizing radiation is administered before one of ZD6474 and one of: 5FU; CPT-11; and 5-FU and CPT-11, or after one of ZD6474 and one of: 5-FU; CPT-11; and 5-FU and CPT-11.

According to one aspect of the present invention the ionizing radiation is administered before any of ZD6474 and one of: 5-FU; CPT-11; and 5-FU and CPT-11 or after all of ZD6474 and one of: 5-FU; CPT-11; and 5-FU and CPT-11.

According to one aspect of the present invention, the ZD6474 is administered to a warm-blooded animal after the animal has been treated with ionizing radiation.

As indicated above, the combination treatments of the present invention, which consist of ZD6474, optionally with ionizing radiation, combined with one of: 5-FU; CPT-11; and 5-FU and CPT-11, as defined herein, are of interest for their antiangiogenic effects and / or their effects on vascular permeability. Angiogenesis and / or increased vascular permeability are present in a wide range of diseases including cancer (including leukemia, multiple myeloma and lymphoma), diabetes, psoriasis, rheumatoid arthritis, Kaposi's sarcoma, hemangioma, acute and chronic kidney disease, atheroma , arterial restenosis, autoimmune diseases, acute inflammation, lymphoedema, excessive scarring and adhesion formation, endometriosis, dysfunctional uterine bleeding and ocular diseases with proliferation of retinal vessels (including age-related macular degeneration). Combination treatments of the present invention are expected to be particularly useful in the prophylaxis and treatment of diseases such as cancer and Kaposi's sarcoma. In particular, the combination treatments of the invention are expected to advantageously slow the growth of primary and recurrent solid tumors, for example, colon, breast, prostate, lung and skin tumors. More particularly, such combination treatments of the invention are expected to inhibit any form of cancer associated with VEGF including leukemia, multiple myeloma and lymphoma and also, for example, that inhibit the growth of primary and recurrent solid tumors that are associated with VEGF. , especially tumors that depend significantly on VEGF for growth and dispersion, including, for example, certain tumors of the colon, breast, prostate, lung, vulva and skin.

According to one aspect of the present invention, such combination treatments of the invention are expected to advantageously slow the growth of primary and secondary (recurrent) tumors in colorectal cancer.

In another aspect of the present invention it is expected that the ZD6474, optionally with ionizing radiation, and one of: 5-FU; CPT-11; and 5-FU and CPT-11 inhibit the growth of primary and recurrent solid tumors that are associated with VEGF especially tumors that depend significantly on VEGF for growth and dispersion.

In another aspect of the present invention it is expected that the ZD6474, optionally with ionizing radiation, and one of: 5-FU; CPT-11; and 5-FU and CPT-11 inhibit the growth of primary and recurrent solid tumors that are associated with both VEGF and EGF, especially tumors that depend significantly on VEGF and EGF for growth and dispersion.

According to another aspect of the present invention the effect of a combination of treatment of the present invention is expected to be at least equivalent to the sum of the effects of each of the components of said treatment used alone, that is, of each of ZD6474, 5-FU, CPT-11 and ionizing radiation used alone.

According to another aspect of the present invention the effect of a treatment combination of the present invention is expected to be greater than the sum of the effects of each of the components of said treatment used alone, that is, of each of ZD6474, 5-FU, CPT-11 and ionizing radiation used alone.

According to another aspect of the present invention the effect of a combination of treatment of the present invention is expected to be a synergistic effect.

According to the present invention a combination treatment is defined as providing a synergistic effect if the effect is therapeutically superior, when measured, for example, by the degree of response, the response rate, the time of disease progression or survival period, which is achieved by dosing one or the other of the components of the combination treatment at its conventional dose. For example, the effect of the combination treatment is synergistic if the effect is therapeutically superior to the effect achievable with ZD6474, 5-FU, CPT-11, 5-FU and CPT-11, or the ionizing radiation used alone. In addition, the effect of the combination treatment is synergistic if a beneficial effect is obtained in a group of patients who do not respond (or respond poorly) to ZD6474, 5-FU, CPT-11, 5-FU and CPT-11, or radiation Ionizing used alone. In addition, the effect of the combination treatment is defined by achieving a synergistic effect if one of the components is dosed at its conventional dose and the other or the other components are dosed at a reduced dose, and the therapeutic effect measured, for example, by the degree of response, the speed of response, the time of progression of the disease or the period of survival, is equivalent to that which can be achieved after the dosing of conventional amounts of the components of the combination treatment. In particular, it is considered that there is synergy if the conventional dose of ZD6474, 5-FU, CPT-11, 5-FU and CPT-11, or ionizing radiation can be reduced without detriment to one or more between the degree of response, the speed of response, the time for disease progression or survival data, in particular without detriment to the degree of response, but with fewer side effects and / or with less problematic side effects than those that occur when used Conventional doses of each component.

The compositions described herein may be in a form suitable for oral administration, for example as tablets or capsules, for nasal administration or inhalation administration, for example as a powder or solution, for parenteral injection (including intravenous injection , subcutaneous, intramuscular, intravascular or perfusion) for example as a sterile solution, suspension or emulsion, for topical administration, for example as an ointment or cream, for rectal administration, for example as a suppository, or the route of administration may be by direct injection into the tumor or by regional administration or by local administration. In other embodiments of the present invention, the combination treatment ZD6474 can be administered endoscopically, intratracheally, intralesionally, percutaneously, intravenously, subcutaneously, intraperitoneally or intratumorally. In general, the compositions described herein can be prepared in a conventional manner using conventional excipients. The compositions of the present invention are advantageously presented in a unit dosage form.

Compound ZD6474 will normally be administered to a warm-blooded animal at a unit dose within the range of 10-500 mg per square meter of the animal's body area, for example approximately 0.3-15 mg / kg in a human being. A unit dose is provided in the range of, for example, 0.3 to 15 mg / kg, preferably 0.5 to 5 mg / kg is contemplated, and this is usually a therapeutically effective dose. A unit dosage form such as a tablet or capsule will usually contain, for example, 25 to 500 mg of active ingredient. Preferably, a daily dose in the range of 0.5-5 mg / kg is used.

The compound CPT-11 is also known as irinotecan. The CPT-11 can be administered according to any known route of administration and dosage.

For example, CPT-11 can be dosed at 350 mg / m2 as an intravenous infusion over a period of 30 to 90 minutes every 3 weeks.

CPT-11 is a semi-synthetic derivative of camptothecin and is metabolized in vivo to an active metabolite SN-38.

The 5-FU compound is 5-fluorouracil. 5-FU can be administered according to any known route of administration and dosage.

For example, 5-FU can be administered as a daily intravenous infusion of 15 mg / kg diluted in 500 ml of 5% dextrose solution or 500 ml of 0.9% sodium chloride solution administered by intravenous infusion: at flow rate of 40 drops per minute in 4 hours; or administered by infusion in 30 to 60 minutes; or as a continuous daily infusion in 24 hours. It is recommended that the daily dose of 5-FU does not exceed 1 g. Usually, 5-FU is administered daily by one of these routes until 12-15 g has been administered and this constitutes a 5-FU course. It is usual practice to leave 4 to 6 weeks between 5-FU courses. Alternatively, 5-FU may be administered by intravenous injection at a dose of 12 mg / kg on three consecutive days, followed by 6 mg / kg on days 5, 7 and 9, that is, the next three alternate days, followed by a Maintenance dose of 5-15 mg / kg by intravenous injection once a week. Alternatively, 5-FU can be administered by intravenous injection at a dose of 15 mg / kg once a week for the duration of the patient's treatment. 5-FU can also be dosed intra-arterially as a regional infusion at 5-7.5 mg / kg by a continuous 24-hour infusion. 5-FU can also be administered orally at a dose of 15 mg / kg once a week or at a dose of 15 mg / kg for six successive days followed by 15 mg / kg once a week.

5-FU is commonly administered with leucovorin. For the avoidance of doubt, the combination treatments of the present invention include the use of 5-FU when given with or without leucovorin.

Leucovorin can be administered according to any known route of administration and dosage. For example, leucovorin can be administered orally. When used in combination with 5-FU, leucovorin is conveniently administered as calcium leucovorin and given intravenously. For example, calcium leucovorin can be administered at a dose of 200 mg / m2 by slow intravenous injection, followed immediately by 5FU at an initial dose of 370 mg / m2 by intravenous injection. Leucovorin injection should not be given more quickly than in 3-5 minutes due to the calcium content of the solution. This treatment is repeated daily for 5 consecutive days. Subsequent courses can be administered after an untreated interval of 21-28 days.

Alternatively, the following regimen can be used: 500 mg / m2 of leucovorin administered by infusion in 2 hours every week for 6 weeks with 500 mg / m2 of 5-FU administered as a bolus via IV. halfway through the 6 week period.

Alternatively, the following regimen can be used: 200 mg / m2 of leucovorin administered by infusion i.v. in 2 hours followed by 400 mg / m2 of 5-FU in bolus i.v. followed by 600 mg / m2 of 5-FU administered by infusion

i.v. in 22 hours, repeated for 2 consecutive days. The cycle is repeated every 2 weeks.

Alternatively, 5-FU can be administered orally as capecitabine (Xeloda ™), tegafur, or TS-1. Capecitabine is a relatively non-cytotoxic fluoropyrimidine carbamate that functions as an orally administered 5FU precursor. Capecitabine can be administered according to any known dosage. For example, a dose of 1250 mg / m2 can be administered orally twice a day, (equivalent to a daily dose of 2500 mg / m2), for 14 days followed by a rest period of 7 days.

Combination treatments of the present invention include the use of 5-FU when administered in any form, (including prodrug and precursor forms that become 5-FU systemically or within the tumor), when administered by any route. and when administered with or without leucovorin.

Combination treatments of the present invention include the use of CPT-11 or SN-38 when administered in any form, (including prodrug and precursor forms that become SN-38 systemically or within the tumor and including liposomial formulations ), when administered by any route.

Radiation therapy can be administered according to known practices in clinical radiotherapy. The dosages of ionizing radiation will be those known for use in clinical radiotherapy. The radiation therapy used will include for example the use of γ rays, X-rays, and / or the directed administration of radiation from radioisotopes. Other forms of DNA damaging factors are also included in the present invention, such as microwaves and UV irradiation. For example, X-rays can be dosed in daily doses of 1.8 to 2.0 Gy, 5 days per week for 5 to 6 weeks. Normally a total fractional dose will be in the range of 45 to 60 Gy. Larger single doses, for example 5 to 10 Gy, can be administered as part of a radiotherapy course. Single doses can be administered intraoperatively. Hyperfractionated radiation therapy can be used, whereby small doses of X-rays are regularly administered over a period of time, for example 0.1 Gy per hour over several days. The dosage ranges for radioisotopes vary widely, and depend on the half-life of the isotope, the strength and type of radiation emitted, and the absorption by the cells.

As indicated above, the dose size of each therapy that is required for the therapeutic or prophylactic treatment of a particular disease will necessarily vary depending on the host treated, the route of administration and the severity of the disease being treated. . Therefore, the optimal dosage can be determined by the doctor who is treating a particular patient. For example, it may be necessary or desirable to reduce the aforementioned doses of the components of the combination treatments, in order to reduce the toxicity.

The combination treatments of the present invention comprise: ZD6474 and 5-FU; ZD6474 and CPT-11; ZD6474, 5-FU and CPT-11; ZD6474, 5-FU and ionizing radiation; ZD6474, CPT-11 and ionizing radiation; ZD6474, 5-FU, CPT-11 and ionizing radiation. The agents thereof can be administered separately or sequentially in any order, or they can be administered simultaneously.

The present invention comprises combinations of 5-FU or CPT-11 or 5-FU and CPT-11 with ZD6474 or with a salt of ZD6474.

The salts for use in the pharmaceutical compositions will be pharmaceutically acceptable salts, but other salts may be useful in the production of ZD6474 and its pharmaceutically acceptable salts. Such salts can be formed with an organic or inorganic base that provides a pharmaceutically acceptable cation. These salts with inorganic or organic bases include, for example, an alkali metal salt, such as a sodium or potassium salt, an alkaline earth metal salt such as a calcium or magnesium salt, an ammonium salt or , for example, a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris- (2-hydroxyethyl) amine.

ZD6474 can be prepared, for example, according to any of the following procedures illustrated by examples (a) - (c) in which, unless otherwise indicated: -

(i)

 evaporations were carried out by vacuum rotary evaporation and the final treatment procedures were carried out after removing the residual solids, such as drying agents, by filtration;

(ii)

 operations were carried out at room temperature, that is, in the range of 18-25 ° C, and in an atmosphere of an inert gas such as argon;

(iii) column chromatography (by rapid procedure) and medium pressure liquid chromatography (MPLC) were performed on Merck's Kieselgel silica (Art. 9385) or Merck's Lichroprep RP-18 reverse phase silica (Art 9303) obtained from E. Merck, Darmstadt, Germany;

(iv)

 yields are provided for illustration only and are not necessarily the maximum that can be achieved;

(v)

 the melting points are not corrected and were determined using an automatic Mettler SP62 melting point apparatus, an oil bath apparatus or a Koffler hot plate apparatus.

(saw)

 the structures of the final products of formula I were confirmed by nuclear magnetic resonance (NMR) (usually proton) and mass spectroscopy techniques; The values of the proton magnetic resonance chemical shifts were determined on the delta scale and the multiplicities of the peaks are shown as follows: s, singlet; d, double up; t, triplet; m, multiplet; br, width; q, quadruplet; NMR spectra were performed in a 400 MHz apparatus at 24 ° C.

(vii) the intermediates generally were not fully characterized and purity was evaluated by thin layer chromatography (TLC), high resolution liquid chromatography (HPLC), infra-red (IR) analysis or NMR analysis;

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(viii) the following abbreviations were used:

DMF N, N-dimethylformamide

DMSO dimethylsulfoxide

THF tetrahydrofuran

TFA trifluoroacetic acid

NMP 1-methyl-2-pyrrolidinone.]

Procedure (a)

A solution of 37% aqueous formaldehyde (50 µl, 0.6 mmol) followed by sodium cyanoborohydride (23 mg, 0.36 mmol) was added to a solution of 4- (4-bromo-2-fluoroanilino) -6 -methoxy-7- (piperidin-4-ylmethoxy) quinazoline (139 mg, 0.3 mmol), in a mixture of THF / methanol (1.4 ml / 1.4 ml). After stirring for 1 hour at room temperature, water was added and volatile components were removed in vacuo. The residue was triturated with water, filtered, washed with water, and dried in vacuo. The solid was purified by chromatography on neutral alumina eluting with methylene chloride followed by methylene chloride / ethyl acetate (1/1) followed by methylene chloride / ethyl acetate / methanol (50/45/5). Fractions containing the expected product were evaporated in vacuo. The resulting white solid was dissolved in methylene chloride / methanol (3 ml / 3 ml) and 3 N hydrogen chloride in ether (0.5 ml) was added. Volatile components were removed in vacuo. The solid was triturated with ether, filtered, washed with ether and dried in vacuo to give 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) hydrochloride. ) quinazoline (120 mg, 69%).

EM-ESI: 475-477 [MH] +

The NMR spectrum of the protonated form of 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4ylmethoxy) quinazoline hydrochloride shows the presence of 2 forms A and B in an A ratio: B of about 9: 1.

1H NMR Spectrum: (DMSOd6; CF3COOD) 1.55-1.7 (m, form A 2H); 1.85-2.0 (m, form B 4H); 2.03 (d, form A 2H); 2.08-2.14 (br s, form A 1H); 2.31-2.38 (br s, form B 1H); 2.79 (s, form A 3H); 2.82 (s, form B 3H); 3.03 (t, form A 2H); 3.21 (br s, form B 2H); 3.30 (br s, form B 2H); 3.52 (d, form A 2H); 4.02 (s, 3 H); 4.12 (d, form A 2H); 4.30 (d, form B 2H); 7.41 (s, 1 H); 7.5-7.65 (m, 2H); 7.81 (d, 1 H); 8.20 (s, 1 H); 8.88 (s, 1 H)

Elemental analysis: Found C 46.0 H 5.2 N 9.6

C22H24N4O2BrF 0.3H2O 2.65H Theoretical C 45.8 H 4.8 N 9.7%

The starting material was prepared as follows:

A solution of 7-benzyloxy-4-chloro-6-methoxyquinazoline hydrochloride (8.35 g, 27.8 mmol), (prepared, for example, as described in WO 97/22596, example 1), and 4-Bromo-2-fluoroaniline (5.65 g, 29.7 mmol) in 2-propanol (200 ml) was heated at reflux for 4 hours. The resulting precipitate was collected by filtration, washed with 2-propanol and then with ether and dried in vacuo to give 7-benzyloxy-4- (4-bromo-2-fluoroanilino) -6-methoxyquinazoline hydrochloride (9.46 g , 78%).

1H NMR Spectrum: (DMSOd6; CD3COOD) 4.0 (s, 3H); 5.37 (s, 2 H); 7.35-7.5 (m, 4H); 7.52-7.62 (m, 4H); 7.8 (d, 1 H); 8.14 (9s, 1H); 8.79 (s, 1 H)

ESI-MS: 456 [MH] +.

Elemental analysis: Found C 54.0 H 3.7 N 8.7

C22H17N3O2BrF 0.9HCl Theoretical C 54.2 H 3.7 N 8.6%

A solution of 7-benzyloxy-4- (4-bromo-2-fluoroanilino) -6-methoxyquinazoline hydrochloride (9.4 g, 19.1 mmol) in TFA (90 ml) was heated at reflux for 50 minutes. The mixture was allowed to cool and poured onto ice. The precipitate

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Four. Five

The resulting was collected by filtration and dissolved in methanol (70 ml). The solution was adjusted to pH 9-10 with concentrated aqueous ammonia solution. The mixture was concentrated to half of the initial volume by evaporation. The resulting precipitate was collected by filtration, washed with water and then with ether, and dried in vacuo to give 4 (4-bromo-2-fluoroanilino) -7-hydroxy-6-methoxyquinazoline (5.66 g, 82 %).

1H NMR Spectrum: (DMSOd6; CD3COOD) 3.95 (s, 3H); 7.09 (s, 1 H); 7.48 (s, 1 H); 7.54 (t, 1 H); 7.64 (d, 1 H); 7.79 (s, 1 H); 8.31 (s, 1 H)

ESI-MS: 366 [MH] +.

Elemental analysis: Found C 49.5 H 3.1 N 11.3

C15H11N3O2BrF Theoretical C49.5 H 3.0 N 11.5%

Maintaining the temperature in the range of 0-5 ° C, a solution of di-tert-butyl dicarbonate (41.7 g, 0.19 mol) in ethyl acetate (75 ml) was added portionwise to a solution of Ethyl 4-piperidinecarboxylate (30 g, 0.19 mol) in ethyl acetate (150 ml) cooled to 5 ° C. After stirring for 48 hours at room temperature, the mixture was poured into water (300 ml). The organic layer was separated, washed successively with water (200 ml), 0.1 N aqueous hydrochloric acid (200 ml), saturated sodium hydrogen carbonate (200 ml) and brine (200 ml), dried (MgSO4) and dried. evaporated to give ethyl 4- (1- (tert-butoxycarbonyl) piperidine) carboxylate (48 g, 98%).

1H NMR Spectrum: (CDCl3) 1.25 (t, 3H); 1.45 (s, 9H); 1.55-1.70 (m, 2H); 1.8-2.0 (d, 2H); 2.35-2.5 (m, 1 H); 2.7-2.95 (t, 2H); 3.9-4.1 (br s, 2H); 4.15 (q, 2H)

A solution of 1 M lithium aluminum hydride in THF (133 ml, 0.133 mol) was added portionwise to a solution of ethyl 4- (1- (tert-butoxycarbonyl) piperidine) carboxylate (48 g, 0.19 mol ) in dry THF (180 ml) cooled to 0 ° C. After stirring at 0 ° C for 2 hours, water (30 ml) was added followed by 2 N sodium hydroxide (10 ml). The precipitate was filtered off through diatomaceous earth and washed with ethyl acetate. The precipitate was washed with water, brine, dried (MgSO4) and evaporated to give 1- (tert-butoxycarbonyl) -4hydroxymethylpiperidine (36.3 g, 89%).

MS (EI): 215 [M.] +

1H NMR Spectrum: (CDCl3) 1.05-1.2 (m, 2H); 1.35-1.55 (m, 10H); 1.6-1.8 (m, 2H); 2.6-2.8 (t, 2H); 3.4-3.6 (t, 2H); 4.04.2 (br s, 2H)

1,4-Diazabicyclo [2.2.2] octane (42.4 g, 0.378 mol) was added to a solution of 1- (tert-butoxycarbonyl) -4-hydroxymethylpiperidine (52.5 g, 0.244 mol) in tert-butyl methyl -ether (525 ml). After stirring for 15 minutes at room temperature, the mixture was cooled to 5 ° C and a solution of toluenesulfonyl chloride (62.8 g, 0.33 mmol) in tert-butyl methyl ether (525 ml) was added in portions for 2 hours while maintaining the temperature at 0 ° C. After stirring at room temperature for 1 hour, petroleum ether (1 L) was added. The precipitate was filtered off. The filtrate was evaporated to give a solid. The solid was dissolved in ether and washed successively with 0.5 N aqueous hydrochloric acid (2 x 500 ml), water, saturated sodium hydrogen carbonate and brine, dried (MgSO4) and evaporated to give 1- (tert) butoxycarbonyl) -4- (4-methylphenylsulfonyloxymethyl) piperidine (76.7 g, 85%).

MS (ESI): 392 [MNa] +

1H NMR Spectrum: (CDCl3) 1.0-1.2 (m, 2H); 1.45 (s, 9H); 1.65 (d, 2H); 1.75-1.9 (m, 2H); 2.45 (s, 3 H); 2.55-2.75 (m, 2H); 3.85 (d, 1 H); 4.0-4.2 (br s, 2H); 7.35 (d, 2H); 7.8 (d, 2H)

Potassium carbonate (414 mg, 3 mmol) was added to a suspension of 4- (4-bromo-2-fluoroanilino) -7-hydroxy-6-methoxyquinazoline (546 mg, 1.5 mmol) in DMF (5 ml). After stirring for 10 minutes at room temperature, 1- (tert-butoxycarbonyl) -4- (4-methylphenylsulfonyloxymethyl) piperidine (636 mg, 1.72 mmol) was added and the mixture was heated at 95 ° C for 2 hours. After cooling, the mixture was poured onto chilled water (20 ml). The precipitate was collected by filtration, washed with water, and dried under vacuum to give 4- (4-bromo-2-fluoroanilino) -7- (1- (tert-butoxycarbonyl) piperidin-4-ylmethoxy) -6-methoxyquinazoline (665 mg, 79%).

EM-ESI: 561-563 [MH] +

1H NMR Spectrum: (DMSOd6) 1.15-1.3 (m, 2H), 1.46 (s, 9H), 1.8 (d, 2H), 2.0-2.1 (m, 1H) , 2.65-2.9 (m, 2H), 3.95 (s, 3H), 4.02 (br s, 2H), 4.05 (d, 2H), 7.2 (s, 1H) , 7.48 (d, 1H), 7.55 (t, 1H), 7.65 (d, 1H), 7.8 (s, 1H), 8.35 (s, 1H), 9.55 ( br s, 1H)

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TFA (3 ml) was added to a suspension of 4- (4-bromo-2-fluoroanilino) -7- (1- (tert-butoxycarbonyl) piperidin-4-ylmethoxy) 6-methoxyquinazoline (673 mg, 1.2 mmol ) in methylene chloride (10 ml). After stirring for 1 hour at room temperature, volatile components were removed in vacuo. The residue was triturated with a mixture of water / ether. The organic layer was separated. The aqueous layer was washed again with ether. The aqueous layer was adjusted to pH 10 with 2 N aqueous sodium hydroxide. The aqueous layer was extracted with methylene chloride. The organic layer was dried (MgSO4) and the solvent was removed in vacuo. The solid was triturated with a mixture of petroleum ether / ether (1/1), filtered, washed with ether and dried in vacuo to give 4- (4-bromo-2-fluoroanilino) -6-methoxy- 7- (piperidin-4-ylmethoxy) quinazoline (390 mg, 70.5%).

ESI-EM: 461-463 [MH] +

1H NMR Spectrum: (DMSOd6) 1.13-1.3 (m, 2H), 1.75 (d, 2H), 1.87-2.0 (m, 1H), 2.5 (d, 2H) , 3.0 (d, 2H), 3.96 (s, 3H), 3.98 (d, 2H), 7.2 (s, 1H), 7.5 (dd, 1H), 7.55 ( t, 1H), 7.68 (dd, 1H), 7.80 (s, 1H), 8.36 (s, 1H), 9.55 (br s, 1H)

Elemental analysis: Found C 54.5 H 4.9 N 12.1

C21H22N4O2BrF Theoretical C 54.7 H 4.8 N 12.1%

Procedure (b)

37% aqueous formaldehyde (3.5 ml, 42 mmol) was added to a solution of 4- (4-bromo-2-fluoroanilino) -7- (1- (tert-butoxycarbonyl) piperidin-4-ylmethoxy) -6-methoxyquinazoline (3.49 g, 6.22 mmol), (prepared as described for the starting material in the procedure (a) above), in formic acid (35 ml). After heating at 95 ° C for 4 hours the volatile components were removed in vacuo. The residue was suspended in water and the mixture was adjusted to pH 10.5 by the slow addition of a 2 N solution of sodium hydroxide. The suspension was extracted with ethyl acetate. The organic layer was washed with brine, dried (MgSO4) and evaporated to give 4- (4-bromo-2-fluoroanilino) 6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline (2.61 g, 88%).

EM-ESI: 475-477 [MH] +

1H NMR Spectrum: (DMSOd6) 1.3-1.45 (m, 2H), 1.8 (d, 2H), 1.7-1.9 (m, 1H), 1.95 (t, 2H) , 2.2 (s, 3H), 2.85 (d, 2H), 3.96 (s, 3H), 4.05 (d, 2H), 7.19 (s, 1H), 7.5 ( d, 1H), 7.55 (t, 1H), 7.67 (d, 1H), 7.81 (s, 1H), 8.37 (s, 1H), 9.54 (s, 1H)

Elemental analysis: Found C 55.4 H 5.1 N 11.6

C22H24N4O2BrF Theoretical C 55.6 H 5.1 N 11.8%

Procedure (c)

A suspension of 4-chloro-6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline (200 mg, 0.62 mmol) and 4-bromo-2-fluoroaniline was heated at reflux for 1.5 hours (142 mg, 0.74 mmol) in isopropanol (3 ml) containing 6 N hydrogen chloride in isopropanol (110 µl, 0.68 ml). After cooling, the precipitate was collected by filtration, washed with isopropanol followed by ether and dried under vacuum to give 4- (4-bromo-2-fluoroanilino) -6-methoxy7- (1-methylpiperidine-4) hydrochloride -ylmethoxy) quinazoline (304 mg, 90%).

Elemental analysis: Found C 47.9 H 4.9 N 10.0

C22H24N4O2BrF 0.5H2O 1.8HCl Theoretical C 48.2 H 5.0 N 10.1%

0.08 isopropanol

The NMR spectrum of the protonated form of 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4ylmethoxy) quinazoline hydrochloride shows the presence of two forms A and B in an A ratio: B of about 9: 1.

1H NMR Spectrum: (DMSOd6) 1.6-1.78 (m, form A 2H); 1.81-1.93 (br s, form B 4H); 1.94-2.07 (d, form A 2H); 2.08-2.23 (br s, form A 1H); 2.29-2.37 (br s, form B 1H); 2.73 (d, form A 3H); 2.77 (d, form B 3H); 2.93-3.10 (q, form A 2H); 3.21 (br s, form B 2H); 3.27 (br s, form B 2H); 3.42-3.48 (d, form A 2H); 4.04 (s, 3 H); 4.10 (d, form

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At 2H); 4.29 (d, form B 2H); 7.49 (s, 1 H); 7.53-7.61 (m, 2H); 7.78 (d, 1 H); 8.47 (s, 1 H); 8.81 (s, 1 H); 10.48 (br s, form A 1H); 10.79 (br s, form B 1H); 11.90 (br s, 1H)

For another NMR reading, some solid potassium carbonate was added to the DMSO solution of 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline hydrochloride described above, in order to release the free base in the NMR tube. The NMR spectrum was then recorded again and presented only one form as described below:

1H NMR Spectrum: (DMSOd6; solid potassium carbonate) 1.3-1.45 (m, 2H); 1.75 (d, 2H); 1.7-1.9 (m, 1 H); 1.89 (t, 2H); 2.18 (s, 3 H); 2.8 (d, 2H); 3.98 (s, 3 H); 4.0 (d, 2H); 7.2 (s, 1 H); 7.48 (d, 1 H); 7.55 (t, 1 H); 7.68 (d, 1 H); 7.8 (s, 1 H); 8.35 (s, 1 H); 9.75 (s, 1 H)

A sample of 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline (free base) was generated from 4- (4-bromo-2 hydrochloride -fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline, (prepared as described above), as follows:

4- (4-Bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxyquinazoline hydrochloride (50 mg) in methylene chloride (2 ml) was suspended and washed with sodium hydrogen carbonate The solution of methylene chloride was dried (MgSO4) and volatile components were removed by evaporation to give 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methlylpiperidin-4-ylmethoxy) quinazoline (free base) The NMR of the free base thus generated has only one form as described below:

1H NMR Spectrum: (DMSOd6) 1.3-1.45 (m, 2H); 1.76 (d, 2H); 1.7-1.9 (m, 1 H); 1.9 (t, 2H); 2.19 (s, 3 H); 2.8 (d, 2H); 3.95 (s, 3 H); 4.02 (d, 2H); 7.2 (s, 1 H); 7.48 (d, 1 H); 7.55 (t, 1 H); 7.68 (dd, 1 H); 7.8 (s, 1 H); 8.38 (s, 1 H); 9.55 (br s, 1H)

For another NMR reading, some CF3COOD was added to the NMR DMSO solution of 4- (4-bromo-2fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline (free base) described above and the NMR spectrum was recorded again. The spectrum of the protonated form of the trifluoroacetate salt of 4- (4-bromo-2-fluoroanilino) -6-methoxy-7 (1-methylpiperidin-4-ylmethoxy) quinazoline thus obtained shows the presence of two forms A and B in an A: B ratio of approximately 9: 1.

1H NMR Spectrum: (DMSOd6; CF3COOD) 1.5-1.7 (m, form A 2H); 1.93 (br s, form B 4H); 2.0-2.1 (d, form A 2H); 2.17 (br s, form A 1H); 2.35 (br s, form B 1H); 2.71 (s, form A 3H); 2.73 (s, form B 3H); 2.97-3.09 (t, form A 2H); 3.23 (br s, form B 2H); 3.34 (br s, form B 2H); 3.47-3.57 (d, form A 2H); 4.02 (s, 3 H); 4.15 (d, form A 2H); 4.30 (d, form B 2H); 7.2 (s, 1 H); 7.3-7.5 (m, 2H); 7.6 (d, 1 H); 7.9 (s, 1 H); 8.7 (s, 1 H)

The starting material was prepared as follows:

1- (tert-Butoxycarbonyl) -4- (4-methylphenylsulfonyloxymethyl) piperidine (40 g, 0.11 mol), (prepared as described for the starting material in the procedure (a) above), was added to a suspension of ethyl 4-hydroxy-3-methoxybenzoate (19.6 g, 0.1 mol) and potassium carbonate (28 g, 0.2 mol) in dry DMF (200 ml). After stirring at 95 ° C for 2.5 hours, the mixture was cooled to room temperature and partitioned between water and ethyl acetate / ether. The organic layer was washed with water, brine, dried (MgSO4) and evaporated. The resulting oil was crystallized from petroleum ether and the suspension was stored overnight at 5 ° C. The solid was collected by filtration, washed with petroleum ether and dried in vacuo to give ethyl 4- (1- (tert-butoxycarbonyl) piperidin-4-ylmethoxy) 3-methoxybenzoate (35 g, 89%).

m.p. 81-83 ° C

MS (ESI :): 416 [MNa] +

1H NMR Spectrum: (CDCl3) 1.2-1.35 (m, 2H); 1.4 (t, 3H); 1.48 (s, 9H); 1.8-1.9 (d, 2H); 2.0-2.15 (m, 2H); 2.75 (t, 2H); 3.9 (d, 2H); 3.95 (s, 3 H); 4.05-4.25 (br s, 2H); 4.35 (q, 2H); 6.85 (d, 1 H); 7.55 (s, 1 H); 7.65 (d, 1 H)

Elemental analysis: Found C 63.4 H 8.0 N 3.5

C21H31NO6 0.3H2O Theoretical C 63.2 H 8.0 N 3.5%

Formaldehyde (12 M, 37% in water, 35 ml, 420 mmol) was added to a solution of ethyl 4- (1- (tert-butoxycarbonyl) piperidin-4-ylmethoxy) -3-methoxybenzoate (35 g, 89 mmol) in formic acid (35 ml). After stirring at 95 ° C for 3 hours, volatile components were removed by evaporation. The residue was dissolved in methylene chloride and 3M hydrogen chloride in ether (40 ml, 120 mmol) was added. After dilution with ether, the mixture was triturated until a solid formed. The solid was collected by filtration, washed with ether and dried in vacuo overnight at 50 ° C to give ethyl 3-methoxy-4- (1-methylpiperidin-4-ylmethoxy) benzoate (30.6 g, quantitative).

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MS (ESI): 308 [MH] + +

1H NMR Spectrum: (DMSOd6) 1.29 (t, 3H); 1.5-1.7 (m, 2H); 1.95 (d, 2H); 2.0-2.15 (br s, 1 H); 2.72 (s, 3 H); 2.9-3.1 (m, 2H); 3.35-3.5 (br s, 2H); 3.85 (s, 3 H); 3.9-4.05 (br s, 2H); 4.3 (q, 2H); 7.1 (d, 1 H); 7.48 (s, 1 H); 7.6 (d, 1H)

A solution of ethyl 3-methoxy-4- (1-methylpiperidin-4-ylmethoxy) benzoate (30.6 g, 89 mmol) in methylene chloride (75 ml) was cooled to 0-5 ° C. TFA (37.5 ml) was added followed by dropwise addition for 15 minutes of a solution of 24 N smoking nitric acid (7.42 ml, 178 mmol) in methylene chloride (15 ml). After completing the addition, the solution was allowed to warm and stirred at room temperature for 2 hours. Volatile components were removed in vacuo and the residue was dissolved in methylene chloride (50 ml). The solution was cooled to 0-5 ° C and ether was added. The precipitate was collected by filtration, and dried under vacuum at 50 ° C. The solid was dissolved in methylene chloride (500 ml) and 3M hydrogen chloride in ether (30 ml) was added followed by ether (500 ml). The solid was collected by filtration and dried in vacuo at 50 ° C to give ethyl 3-methoxy-4- (1-methylpiperidin-4-ylmethoxy) -6-nitrobenzoate (28.4 g, 82%).

MS (ESI): 353 [MH] +.

1H NMR Spectrum: (DMSOd6) 1.3 (t, 3H); 1.45-1.65 (m, 2H); 1.75-2.1 (m, 3H); 2.75 (s, 3 H); 2.9-3.05 (m, 2H); 3.4-3.5 (d, 2H); 3.95 (s, 3 H); 4.05 (d, 2H); 4.3 (q, 2H); 7.32 (s, 1 H); 7.66 (s, 1 H)

A suspension of ethyl 3-methoxy-4- (1-methylpiperidin-4-ylmethoxy) -6-nitrobenzoate (3.89 g, 10 mmol) in methanol (80 ml) containing 10% platinum on activated carbon (50 % wet) (389 mg) was hydrogenated at 1.8 atmospheres of pressure until hydrogen absorption ceased. The mixture was filtered and the filtrate was evaporated. The residue was dissolved in water (30 ml) and adjusted to pH 10 with a saturated solution of sodium hydrogen carbonate. The mixture was diluted with ethyl acetate / ether (1/1) and the organic layer was separated. The aqueous layer was further extracted with ethyl acetate / ether and the organic layers were combined. The organic layers were washed with water, brine, dried (MgSO4), filtered and evaporated. The resulting solid was triturated in a petroleum ether / ether mixture, filtered, washed with petroleum ether and dried in vacuo at 60 ° C to give 6-amino-3-methoxy-4- (1-methylpiperidine -4-methylmethoxy) ethyl benzoate (2.58 g, 80%).

m.p. 111-112 ° C

MS (ESI): 323 [MH] +.

1H NMR Spectrum: (CDCl3) 1.35 (t, 3H); 1.4-1.5 (m, 2H); 1.85 (m, 3 H); 1.95 (t, 2H); 2.29 (s, 3 H); 2.9 (d, 2H); 3.8 (s, 3H); 3.85 (d, 2H); 4.3 (q, 2H); 5.55 (br s, 2H); 6.13 (s, 1 H); 7.33 (s, 1 H)

Elemental analysis: Found C 62.8 H 8.5 N 8.3

C17H26N2O4 0.2H2O Theoretical C 62.6 H 8.2 N 8.6%

A solution of ethyl 6-amino-3-methoxy-4- (1-methylpiperidin-4-ylmethoxy) benzoate (16.1 g, 50 mmol) in 2-methoxyethanol (160 ml) containing formamidine acetate (5.2 g , 50 mmol) was heated at 115 ° C for 2 hours. Formamidine acetate (10.4 g, 100 mmol) was added portionwise every 30 minutes for 4 hours. Heating was prolonged for 30 minutes after the last addition. After cooling, volatile components were removed in vacuo. The solid was dissolved in ethanol (100 ml) and methylene chloride (50 ml). The precipitate was filtered off and the filtrate was concentrated to a final volume of 100 ml. The suspension was cooled to 5 ° C and the solid was collected by filtration, washed with cold ethanol followed by ether and dried in vacuo overnight at 60 ° C to give 6-methoxy-7- (1-methylpiperidin-4- ilmethoxy) -3,4-dihydroquinazolin-4-one (12.7 g, 70%).

MS (ESI): 304 [MH] +.

1H NMR Spectrum: (DMSOd6) 1.25-1.4 (m, 2H); 1.75 (d, 2H); 1.9 (t, 1 H); 1.9 (s, 3H); 2.16 (s, 2H); 2.8 (d, 2H); 3.9 (s, 3H); 4.0 (d, 2H); 7.11 (s, 1 H); 7.44 (s, 1 H); 7.97 (s, 1 H)

A solution of 6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) -3,4-dihydroquinazolin-4-one (2.8 g, 9.24 mmol) in thionyl chloride (28 ml) containing DMF (280 µl) was heated at reflux at 85 ° C for 1 hour. After cooling, volatile components were removed by evaporation. The precipitate was triturated with ether, filtered, washed with ether and dried in vacuo. The solid was dissolved in methylene chloride and saturated aqueous sodium hydrogen carbonate was added. The organic layer was separated, washed with water, brine, dried (MgSO4) and evaporated to give 4-chloro-6methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline (2.9 g, 98% ).

MS (ESI): 322 [MH] +.

1H NMR Spectrum: (DMSOd6) 1.3-1.5 (m, 2H); 1.75-1.9 (m, 3H); 2.0 (t, 1 H); 2.25 (s, 3 H); 2.85 (d, 2H); 4.02 (s, 3 H); 4.12 (d, 2H); 7.41 (s, 1 H); 7.46 (s, 1 H); 8.9 (s, 1 H)

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Alternatively, 6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) -3,4-dihydroquinazolin-4-one can be prepared as follows:

Sodium hydride (1.44 g of a 60% suspension in mineral oil, 36 mmol) was added portionwise for 20 minutes to a solution of 7-benzyloxy-6-methoxy-3,4-dihydroquinazolin-4-one ( 8.46 g, 30 mmol), (prepared, for example, as described in WO 97/22596, example 1), in DMF (70 ml) and the mixture was stirred for 1.5 hours. Chloromethyl pivalate (5.65 g, 37.5 mmol) was added portionwise and the mixture was stirred for 2 hours at room temperature. The mixture was diluted with ethyl acetate (100 ml) and poured onto ice / water (400 ml) and 2 N hydrochloric acid (4 ml). The organic layer was separated and the aqueous layer was extracted with ethyl acetate, the combined extracts were washed with brine, dried (MgSO4) and the solvent was removed by evaporation. The residue was triturated with a mixture of ether and petroleum ether, the solid was collected by filtration and dried in vacuo to give 7-benzyloxy-6-methoxy-3 - ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4 -one (10 g, 84%).

1H NMR Spectrum: (DMSOd6) 1.11 (s, 9H); 3.89 (s, 3 H); 5.3 (s, 2H); 5.9 (s, 2H); 7.27 (s, 1 H); 7.35 (m, 1 H); 7.47 (t, 2H); 7.49 (d, 2H); 7.51 (s, 1 H); 8.34 (s, 1 H)

A mixture of 7-benzyloxy-6-methoxy-3 - ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4-one (7 g, 17.7 mmol) and 10% palladium catalyst on activated carbon (700 mg) in ethyl acetate (250 ml), DMF (50 ml), methanol (50 ml) and acetic acid (0.7 ml) was stirred under nitrogen at atmospheric pressure for 40 minutes. The catalyst was filtered off and the solvent was removed from the filtrate by evaporation. The residue was triturated with ether, collected by filtration and dried in vacuo to give 7-hydroxy-6-methoxy-3 - ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4-one (4.36 g , 80%).

1H NMR Spectrum: (DMSOd6) 1.1 (s, 9H); 3.89 (s, 3 H); 5.89 (s, 2 H); 7.0 (s, 1 H); 7.48 (s, 1 H); 8.5 (s, 1 H)

Triphenylphosphine (1.7 g, 6.5 mmol) under nitrogen was added to a suspension of 7-hydroxy-6-methoxy-3 ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4-one (1.53 g , 5 mmol) in methylene chloride (20 ml), followed by the addition of 1- (tert-butoxycarbonyl) -4- (hydroxymethyl) piperidine (1.29 g, 6 mmol), (prepared as described for the material starting in the procedure (a) above), and by a solution of diethyl azodicarboxylate (1.13 g, 6.5 mmol) in methylene chloride (5 ml). After stirring for 30 minutes at room temperature, the reaction mixture was poured onto a silica column and eluted with ethyl acetate / petroleum ether (1/1 followed by 6/5, 6/4 and 7/3) . Evaporation of the fractions containing the expected product led to an oil that crystallized after trituration with pentane. The solid was collected by filtration and dried under vacuum to give 7- (1- (tertbutoxycarbonyl) piperidin-4-ylmethoxy) -6-methoxy-3 - ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4- one (232 g, 92%).

EM-ESI: 526 [MNa] +

1H NMR Spectrum: (CDCl3) 1.20 (s, 9H), 1.2-1.35 (m, 2H), 1.43 (s, 9H), 1.87 (d, 2H), 2.05 -2.2 (m, 1H), 2.75 (t, 2H), 3.96 (d, 2H), 3.97 (s, 3H), 4.1-4.25 (br s, 2H) , 5.95 (s, 2H), 7.07 (s, 1H), 7.63 (s, 1H), 8.17 (s, 1H)

Elementary Analysis:

Found C 61.8 H 7.5 N 8.3

C26H37N3O7

Theoretical C 62.0 H 7.4 N 8.3%

Be

stirred  a solution  from 7- (1- (tert-butoxycarbonyl) piperidin-4-ylmethoxy) -6-methoxy-3 - ((pivaloyloxy) methyl) -3.4

dihydroquinazolin-4-one (2.32 g, 4.6 mmol) in methylene chloride (23 ml) containing TFA (5 ml) at room temperature for 1 hour. Volatile components were removed in vacuo. The residue was partitioned between ethyl acetate and sodium hydrogen carbonate. The organic solvent was removed in vacuo and the residue was filtered. The precipitate was washed with water, and dried under vacuum. The solid was azeotropically treated with toluene and dried under vacuum to give 6-methoxy-7- (piperidin-4-ylmethoxy) -3 - ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4-one (1, 7 g, 92%).

EM-ESI: 404 [MH] +.

1H NMR Spectrum: (DMSOd6; CF3COOD) 1.15 (s, 9H), 1.45-1.6 (m, 2H), 1.95 (d, 2H), 2.1-2.25 (m, 1H), 2.95 (t, 2H), 3.35 (d, 2H), 3.95 (s, 3H), 4.1 (d, 2H), 5.95 (s, 2H), 7, 23 (s, 1H), 7.54 (s, 1H), 8.45 (s, 1H)

An aqueous solution of 37% formaldehyde (501 µl, 6 mmol) followed by sodium cyanoborohydride (228 mg, 3.6 mmol) was added portionwise to a solution of 6-methoxy-7- (piperidin-4-ylmethoxy) -3 - ((pivaloyloxy) methyl) -3,4dihydroquinazolin-4-one (1.21 g, 3 mmol) in a mixture of THF / methanol (10 ml / 10 ml). After stirring for 30 minutes at room temperature, the organic solvents were removed in vacuo and the residue was partitioned between methylene chloride and water. The organic layer was separated, washed with water and brine, dried (MgSO4) and volatile components were removed by evaporation. The residue was triturated with ether and the resulting solid was collected by filtration, washed with ether and dried in vacuo to give 6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) -3 ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4-one (1.02 g, 82%).

ESI-MS: 418 [MH] +.

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1H NMR Spectrum: (CDCl3) 1.19 (s, 9H), 1.4-1.55 (m, 2H), 1.9 (d, 2H), 2.0 (t, 2H), 1.85 -2.1 (m, 1H), 2.3 (s, 3H), 2.92 (d, 2H), 3.96 (s, 3H), 3.99 (d, 2H), 5.94 ( s, 2H), 7.08 (s, 1H), 7.63 (s, 1H), 8.17 (s, 1H)

A saturated solution of ammonia in methanol (14 ml) was added to a solution of 6-methoxy-7- (1-methylpiperidin-4-methylmethoxy) -3 - ((pivaloyloxy) methyl) -3,4-dihydroquinazolin-4-one ( 1.38 g, 3.3 mmol) in methanol (5 ml). After stirring for 20 hours at room temperature, the suspension was diluted with methylene chloride (10 ml). The solution was filtered. The filtrate was evaporated in vacuo and the residue triturated with ether, collected by filtration, washed with ether and dried in vacuo to give 6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) -3.4 -dihydroquinazolin-4-one (910 mg, 83%).

EM-ESI: 304 [MH] +.

1H NMR Spectrum: (DMSOd6) 1.3-1.45 (m, 2H), 1.75 (d, 2H), 1.7-1.85 (m, 1H), 1.9 (t, 2H) , 2.2 (s, 3H), 2.8 (d, 2H), 3.9 (s, 3H), 4.0 (d, 2H), 7.13 (s, 1H), 7.45 ( s, 1H), 7.99 (s, 1H)

The following tests were used to demonstrate the activity of ZD6474 in combination with 5-FU and CPT-11.

Xenografts of human colon tumors LS-174T in nude mice

107 LS-174T tumor cells in 0.2 ml of serum-free Roswell Park Memorial Institute (RPMI) -1640 medium were injected subcutaneously into the flanks of 10 athymic mice (nu / nu genotype). When the tumor sizes reached 700-1000 mm3 (3-4 weeks), the tumors were surgically removed and the smaller tumor fragments (20-30 mg) were implanted s.c. on the right flank of 120 athymic mice. When the tumors reached a volume of 100 to 200 mm3 (14-16 days after grafting), the mice were divided into random groups (13-15 per group) and treatment was started.

(a) 5-FU + ZD6474

□

 The control group (Group 1) received a daily oral (po) vehicle administration of ZD6474 for 14 consecutive days (days 0 - 13) combined with two intravenous (iv) injections of saline (the vehicle for 5FU) on days 0 and 7.

□

 For Group 2, the treatments consisted of a daily administration p.o. of ZD6474 only at 25 mg / kg / administration for 14 consecutive days (days 0 - 13) combined with two injections i.v. of saline solution (the vehicle for 5-FU) on days 0 and 7. The ZD6474 was prepared as a suspension in 1% polysorbate 80 (that is, a 1% (v / v) solution of polyoxyethylene monooleate (20 ) sorbitan in deionized water).

□

 Group 3 received two injections i.v. 5-FU at 75 mg / kg / injection, on days 0 and 7, combined with a daily administration p.o. ZD6474 vehicle for 14 consecutive days (days 0-13).

□

 Group 4 received daily administration p.o. of ZD6474 at 25 mg / kg / administration for 14 consecutive days (days 0 - 13) combined with two injections i.v. 5-FU at 75 mg / kg / injection, days 0 and 7.

The administration volume of ZD6474 was 10.0 ml / kg (200 µl for a 20 g mouse). The injection volume of 5-FU was 10.0 ml / kg (200 µl for a 20 g mouse).

Group

 Treatments Combined doses of drugs (mg base / kg / iny.) Route of administration No. of treatments No. of treatment / day Interval in days between treatment (days)

one

ZD6474 and 5 vehicles 0.0 p.o. for ZD6474 14 p.o. 1 p.o. 1 for p.o.

FU

vehicle i.v. for saline solution 2 i.v. 1 i.v. 7 for i.v.

2

ZD6474 + saline solution 25.0 p.o. for ZD6474 14 p.o. 1 p.o. 1 for p.o.

i. v. for saline solution

2 i.v. 1 i.v. 7 for i.v.

3

5-FU + ZD6474 vehicle 75.0 i.v. for 5-FU 14 p.o. 1 p.o. 1 for p.o.

p.o. for ZD6474 vehicle

2 i.v. 1 i.v. 7 for i.v.

Group

 Treatments Combined doses of drugs (mg base / kg / iny.) Route of administration No. of treatments No. of treatment / day Interval in days between treatment (days)

4

ZD6474 + 5-FU 25.0 for ZD6474 75.0 for 5-FU p.o. for ZD6474 14 p.o. 1 p.o. 1 for p.o.

i.v. for 5-FU

2 i.v. 1 i.v. 7 for i.v.

Tumor volumes (mm3) were evaluated at least twice a week by means of the bilateral measure by Vernier caliber and, considering the length of the longest tumor diameter and the corresponding perpendicular width, they were calculated using the formula (π / 6) x (length x width) x the square root of (length x width). Growth inhibition from the beginning of treatment was evaluated by comparison of the differences in tumor volumes between the control group and the treated groups. For all mice, the study was stopped when the tumors reached 2,000 mm3. For all mice, tumors were removed and weights were recorded after the end of the study.

image 1

Treatment

Growth tumor inhibition control on day 13 P value (t test for two samples of one tail) Regressions *

ZD6474 (25 mg / kg / day p.o., days 0 -13)

80% 0.01 5/15

5-FU (75 mg / kg IV, days 0 and 7)

68% 0.03 6/15

ZD6474 + 5-FU

107% (Regression) 0.002  8/13

Treatment

Growth tumor inhibition control on day 13 P value (t test for two samples of one tail) Regressions *

* Number of tumors that had suffered a regression ≥ 10% in volume on day 13, when compared with their pre-treatment volume on day 0.

The combination of 5-FU with ZD6474 produced a significantly greater tumor growth inhibition than 5-FU alone (P = 0.018 on day 13, by a t-test of two samples of one tail).

The combination of 5-FU with ZD6474 produced a tumor growth inhibition significantly greater than 5 on ZD6474 alone, (P = 0.027 on day 13, by a t-test of two samples of one tail).

The combination of 5-FU with ZD6474 produced more tumor regressions (62%) than ZD6474 alone (33%) or 5FU alone (40%).

(b) CPT-11 + ZD6474

□ The control group (Group 1) received a daily oral administration (p.o.) of ZD6474 vehicle for 14 days

10 consecutive days (days 0 - 13) combined with two intravenous injections (iv) of saline solution (the vehicle for CPT-11) on days 0 and 7. The control group did not continue after this period, since some of the volumes of the tumors were considered too large (~ 2 cm3).

□ For Group 2, the treatments consisted of a daily administration p.o. of ZD6474 only at 25 mg / kg / administration for 21 consecutive days (days 0-20) combined with three injections i.v. from solution

15 saline (the vehicle for CPT-11) on days 0, 7 and 14. The ZD6474 was prepared as a suspension in 1% polysorbate 80 (that is, a 1% (v / v) solution of polyoxyethylene monooleate ( 20) sorbitan in deionized water).

□ Group 3 received three injections i.v. of CPT-11 at 20 mg / kg / injection, on days 0, 7 and 14, combined with a daily administration p.o. of vehicle from ZD6474 for 21 consecutive days (days 0 - 20).

□ Group 4 received daily administration p.o. of ZD6474 at 25 mg / kg / administration for 21 consecutive days 20 (days 0-20) combined with three injections i.v. of CPT-11 at 20 mg / kg / injection, on days 0, 7 and 14.

The administration volume of ZD6474 was 10.0 ml / kg (200 µl for a 20 g mouse). The injection volume of CPT-11 was 10.0 ml / kg (200 µl for a 20 g mouse).

Group

 Treatments Combined doses of drugs (mg base / kg / iny.) Route of administration No. of treatments No. of treatment / day Interval in days between treatment (days)

one

Vehicles of ZD6474 and 0.0  p.o. for vehicle of 14 p.o. 1 p.o. 1 for p.o.

CPT-11

ZD6474 2 i.v. 1 i.v. 7 for i.v.

i. v. for saline solution

2

ZD6474 + saline solution 25.0  p.o. for ZD6474 21 p.o. 1 p.o. 1 for p.o.

3 i.v.

1 i.v. 7 for i.v.

i. v. for saline solution

3

CPT-11 + 20.0 i.v. for CPT-11 21 p.o. 1 p.o. 1 for p.o.

Group

 Treatments Combined doses of drugs (mg base / kg / iny.) Route of administration No. of treatments No. of treatment / day Interval in days between treatment (days)

ZD6474 vehicle

p.o. for ZD6474 vehicle 3 i.v. 1 i.v. 7 for i.v.

4

ZD6474 + CPT11 25.0 for ZD6474 p.o. for 21 p.o. 1 p.o. 1 for p.o.

20.0 for CPT-11

ZD6474 3 i.v. 1 i.v. 7 for i.v.

i.v. for CPT-11

Tumor volumes (mm3) were evaluated at least twice a week by means of the bilateral measure by Vernier caliber and, considering the length of the longest tumor diameter and the corresponding perpendicular width, they were calculated using the formula (π / 6) x (length x width) x the square root of (length x width). Growth inhibition from the beginning of treatment was evaluated by comparison of the differences in tumor volumes between the control group and the treated groups. For all mice, the study was stopped when the tumors reached 2,000 mm3. For all mice, tumors were removed and weights were recorded after the end of the study.

image 1

10 The mean volumes of tumors on day 20 for mice treated with ZD6474 (25 mg / kg / day po, days 0 20), CPT-11 (20 mg / kg iv, days 0, 7 and 14), or combination thereof, were 475 mm3, 552 mm3 and 336 mm3 respectively.

An analogous experiment can be used to study the combination of ZD6474, 5-FU and CPT-11 in this animal model.

15 An analogous experiment can be used to study the combination of ZD6474, 5-FU, CPT-11 and ionizing radiation in this animal model.

Claims (11)

1. The use of 4- (4-bromo-2-fluoroanilino) -6-methoxy-7- (1-methylpiperidin-4-ylmethoxy) quinazoline, also known as ZD6474: image 1 5 or one of its pharmaceutically acceptable salts and one of: a) 5-FU; 10 b) CPT-11; Y c) 5-FU and CPT-11 in the manufacture of a medicament for the treatment of cancer in a warm-blooded animal such as a being 15 human.

2.

 The use according to claim 1, wherein the cancer involves a solid tumor.

3.

The use of ZD6474 or a pharmaceutically acceptable salt and 5-FU in the manufacture of a medicine

20 for use in the treatment of cancer in a warm-blooded animal such as a human being being treated with ionizing radiation. 4. The use of ZD6474 or one of its pharmaceutically acceptable salts and CPT-11 in the manufacture of a medicament for use in the treatment of cancer in a warm-blooded animal such as a human being who is being treated with ionizing radiation. 5. The use of ZD6474 or one of its pharmaceutically acceptable salts and 5-FU and CPT-11 in the manufacture of a medicament for use in the treatment of cancer in a warm-blooded animal such as a human being being treated with ionizing radiation. 30

6.

 The use according to any one of claims 3 to 5, wherein the cancer involves a solid tumor.

7.

 The use according to any one of the preceding claims, wherein the cancer is colorectal cancer.

A pharmaceutical composition comprising ZD6474 or a pharmaceutically acceptable salt thereof, and 5-FU in association with a pharmaceutically acceptable carrier or excipient. 9. A pharmaceutical composition comprising ZD6474 or a pharmaceutically acceptable salt thereof, and CPT-11 in association with a pharmaceutically acceptable carrier or carrier, 10. A pharmaceutical composition comprising ZD6474 or a pharmaceutically acceptable salt thereof, and 5FU and CPT-11 in association with a pharmaceutically acceptable carrier or excipient.

eleven.

 A kit comprising ZD6474 or a pharmaceutically acceptable salt thereof, and 5-FU.

12.

 A kit comprising ZD6474 or a pharmaceutically acceptable salt thereof, and CPT-11.

13.

 A kit comprising ZD6474 or a pharmaceutically acceptable salt thereof, and 5-FU and CPT-11.