HK1071743A - Salt forms of e-2-methoxy-n-(3-(4-(3-methyl-pyridin-3-yloxy)-phenylamino)-quinazolin-6-yl)-allyl)-acetamide, its preparation and its use against cancer - Google Patents

Description

Salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, their preparation and their use against cancer

Background

The present invention relates to salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide of formula (I):

formula I

The free base form of formula I is described in unexamined U.S. patent application serial No. 09/883,752 filed on 18.6.2001, which is incorporated herein by reference in its entirety. The above application is a co-application with the present application. The free base of formula I is useful for the treatment of hyperproliferative diseases, such as cancer.

The present invention provides succinate and malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

The present invention also provides sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

The present invention further relates to a process for the preparation of sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide. The invention also relates to pharmaceutical compositions containing the sesquisuccinate and di-malonate salts of the compound of formula I. The salts of the invention are useful for the treatment of hyperproliferative diseases, such as cancer, in mammals, especially humans. The invention also relates to methods of administering the salts of formula I to treat hyperproliferative diseases.

Summary of The Invention

The present invention relates to succinate and malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide of formula I:

formula I

In a preferred embodiment, the present invention relates to the sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

The invention also relates to a process for the preparation of the sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, comprising mixing the free base with one of the above salts in the presence of a suitable organic solvent.

The sesquisuccinate and bis-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide have been characterized by elemental analysis.

It has been surprisingly found that the sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide have high crystallinity, i.e. are substantially free of amorphous material. The advantage of such salts is that they provide more reproducible dosing results. The sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide are substantially hydroscopic stable, which alleviates potential problems associated with weight changes of the active ingredient during capsule or tablet preparation.

The present invention also relates to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a succinate or malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide effective in treating abnormal cell growth. In a preferred embodiment, the present invention relates to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of the sesquisuccinate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide and a bis-malonate salt effective in treating abnormal cell growth.

In one embodiment, the abnormal cell growth treated is cancer.

In one embodiment of the invention, the cancer is selected from lung cancer, non-small cell lung (NSCL) cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head and neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small bowel cancer, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, prostate cancer, chronic and acute leukemias, lymphocytoma, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, tumors of the Central Nervous System (CNS), colorectal cancer (CRC), primary CNS lymphoma, tumors of the spinal axis, brain stem glioma, pituitary adenoma, or a combination of one or more of the foregoing cancers.

In a preferred embodiment of the invention, the cancer is selected from the group consisting of breast cancer, colon cancer, ovarian cancer, non-small cell lung (NSCL) cancer, colorectal cancer (CRC), prostate cancer, bladder cancer, rectal cancer, gastric cancer, endometrial cancer, head and neck cancer, and esophageal cancer.

In a more preferred embodiment of the invention, the cancer is selected from renal cell carcinoma, gastric cancer, colon cancer, breast cancer and ovarian cancer.

In a more preferred embodiment, the cancer is selected from colon cancer, breast cancer or ovarian cancer.

Another embodiment of the present invention relates to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a succinate or malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide in combination with an anti-neoplastic agent effective in treating abnormal cell growth, the antineoplastic agent is selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-hormones, and anti-androgens.

Another embodiment of the present invention relates to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a sesquisuccinate or di-malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide effective to treat abnormal cell growth in combination with an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-cancer agents, Anti-hormones and anti-androgens.

Another embodiment of the present invention is directed to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a succinate or malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide in combination with a cytotoxin effective in treating abnormal cell growth.

In a preferred embodiment of the invention, the cytotoxin is Taxol * (paclitaxel).

The invention further relates to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a succinate or malonate salt of formula I effective in treating abnormal cell growth in combination with a compound selected from the group consisting of cyclophosphamide, 5-fluorouracil, floxuridine, gemcitabine, vinblastine, vincristine, daunorubicin, doxorubicin, epirubicin, tamoxifen, methylprednisolone, cisplatin, carboplatin, CPT-11, gemcitabine, paclitaxel and docetaxel.

In a preferred embodiment, the present invention relates to a method of treating abnormal cell growth in a mammal, which method comprises administering to said mammal an amount of a succinate or malonate salt of formula I effective in treating abnormal cell growth in combination with a compound selected from the group consisting of tamoxifen, cisplatin, carboplatin, paclitaxel and docetaxel.

A preferred embodiment of the invention relates to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal a sesquisuccinate or malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide in combination with a cytotoxin in an amount effective to treat abnormal cell growth.

In a preferred embodiment of the invention, the cytotoxin is Taxol * (paclitaxel).

The invention further relates to a method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a sesquisuccinate or di-malonate salt of formula I effective to treat abnormal cell growth in combination with a compound selected from the group consisting of cyclophosphamide, 5-fluorouracil, floxuridine, gemcitabine, vinblastine, vincristine, daunorubicin, doxorubicin, epirubicin, tamoxifen, methylprednisolone, cisplatin, carboplatin, CPT-11, gemcitabine, paclitaxel and docetaxel.

In a preferred embodiment, the present invention relates to a method of treating abnormal cell growth in a mammal, which comprises administering to said mammal an amount of a sesquisuccinate or di-malonate salt of formula I effective to treat abnormal cell growth in combination with a compound selected from the group consisting of tamoxifen, cisplatin, carboplatin, paclitaxel and docetaxel.

The present invention further relates to a pharmaceutical composition for treating abnormal cell growth in a mammal comprising an amount of a succinate or malonate salt of formula I effective to treat abnormal cell growth and a pharmaceutically acceptable carrier.

The invention further relates to a pharmaceutical composition for treating abnormal cell growth in a mammal comprising an amount of a sesquisuccinate or di-malonate salt of formula I effective to treat abnormal cell growth and a pharmaceutically acceptable carrier.

The present invention also relates to a method of treating abnormal cell growth in a mammal, including a human, comprising administering to said mammal an amount of a sesquisuccinate or di-malonate salt of formula I or a solvate or prodrug thereof effective to treat abnormal cell growth. In one embodiment of the method, the abnormal cell growth is a cancer, including, but not limited to, lung cancer, non-small cell lung (NSCL) cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's disease, carcinoma of the esophagus, carcinoma of the small intestine, cancer of the endocrine system, carcinoma of the thyroid gland, carcinoma of the parathyroid gland, carcinoma of the adrenal gland, sarcoma of soft tissue, carcinoma of the urethra, carcinoma of the penis, carcinoma of the prostate, chronic and acute leukemias, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, neoplasms of the Central Nervous System (CNS), colorectal cancer (CRC), primary CNS lymphoma, Pituitary adenoma or a combination of one or more of the foregoing cancers. In another embodiment of the method, the abnormal cell growth is a benign proliferative disease including, but not limited to, psoriasis, benign prostatic hyperplasia or restenosis.

The present invention also relates to a method of treating abnormal cell growth in a mammal, including a human, comprising administering to said mammal an amount of a sesquisuccinate or di-malonate salt of formula I, or a solvate or prodrug thereof, effective in treating abnormal cell growth in combination with an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-hormones, and anti-androgens.

The present invention also relates to a pharmaceutical composition for treating abnormal cell growth in a mammal, including a human, comprising an amount effective to treat abnormal cell growth of a sesquisuccinate or di-malonate salt of formula I or a solvate or prodrug thereof and a pharmaceutically acceptable carrier. In one embodiment of the composition, the abnormal cell growth is a cancer, including, but not limited to, lung cancer, non-small cell lung (NSCL) cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, carcinoma of the fallopian tubes, endometrial cancer, cervical cancer, vaginal cancer, vulval cancer, hodgkin's disease, esophageal cancer, small bowel cancer, cancer of the endocrine system, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, cancer of the urethra, cancer of the penis, prostate cancer, chronic and acute leukemias, lymphocytic neoplasms, bladder cancer, renal or ureter cancer, renal cell carcinoma, renal pelvis cancer, tumors of the Central Nervous System (CNS), colorectal cancer (CRC), primary CNS lymphoma, spinal axis tumors, brain stem glioma, Pituitary adenoma or a combination of one or more of the foregoing cancers. In another embodiment of the pharmaceutical composition, the abnormal cell growth is a benign proliferative disease including, but not limited to, psoriasis, benign prostatic hyperplasia or restenosis.

The present invention also relates to a pharmaceutical composition for treating abnormal cell growth in mammals, including humans, which comprises a succinate or malonate salt or solvate or prodrug thereof of formula I, in an amount effective to treat abnormal cell growth, in combination with a pharmaceutically acceptable carrier and an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-hormones and anti-androgens.

The present invention also relates to a pharmaceutical composition for treating abnormal cell growth in mammals, including humans, which comprises a combination of an abnormal cell growth treating effective amount of a sesquisuccinate or di-malonate salt of formula I or a solvate or prodrug thereof, with a pharmaceutically acceptable carrier and an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-hormones and anti-androgens.

The invention also relates to a method of treating a mammal suffering from a cancer characterized by overexpression of erbB2 comprising administering to the mammal an amount of a succinate or malonate salt of formula I effective to treat said cancer characterized by overexpression of erbB 2.

A preferred embodiment of the invention relates to a method of treating a mammal suffering from a cancer characterized by overexpression of erbB2 comprising administering to the mammal an amount of a sesquisuccinate or di-malonate salt of formula I effective to treat said cancer characterized by overexpression of erbB 2.

The invention also relates to a method of treating a mammal suffering from a disease characterized by overexpression of erbB2 comprising administering to the mammal an amount of a succinate or malonate salt of formula I effective to treat said disease characterized by overexpression of erbB 2.

A preferred embodiment of the invention relates to a method of treating a mammal suffering from a disease characterized by overexpression of erbB2 comprising administering to the mammal an amount of a sesquisuccinate or di-malonate salt of formula I effective to treat said disease characterized by overexpression of erbB 2.

The invention also relates to a method of inducing cell death comprising contacting a cell which overexpresses erbB2 with an effective amount of a succinate or malonate salt of formula I. In one embodiment, the cell is a mammalian, preferably human, cancer cell.

A preferred embodiment of the invention relates to a method of inducing cell death comprising contacting a cell overexpressing erbB2 with an effective amount of a sesquisuccinate or di-malonate salt of formula I. In one embodiment, the cell is a mammalian, preferably human, cancer cell.

The present invention relates to a method of inducing cell death comprising contacting a cell overexpressing erbB2 with an effective amount of a succinate or malonate salt of formula I, said method further comprising contacting the cell with a growth inhibitory agent.

In another embodiment, the invention relates to a method of inducing cell death comprising contacting a cell overexpressing erbB2 with an effective amount of a sesquisuccinate or di-malonate salt of formula I, said method further comprising contacting the cell with a growth inhibitory agent.

In a preferred embodiment, the cells are exposed to a chemotherapeutic agent or radiation.

The invention further relates to a method of treating cancer in a human, wherein the cancer expresses the erbB2 receptor, which method comprises administering to the human a therapeutically effective amount of a succinate or malonate salt of formula I. In a preferred embodiment, the invention relates to a method of treating cancer in a human, wherein the cancer expresses erbB2 receptor, the method comprising administering to the human a therapeutically effective amount of a sesquisuccinate or di-malonate salt of formula I. In a preferred embodiment of the invention, the cancer is not characterized by overexpression of the erbB1 receptor. In another preferred embodiment, the cancer is characterized by overexpression of the erbB1 and erbB2 receptors.

The invention also relates to a method of treatment of a disease associated with angiogenesis in a mammal, including a human, comprising administering to said mammal an amount of a succinate or malonate salt of formula I, or a solvate or prodrug thereof, effective in treating said disease. In a preferred embodiment, the present invention relates to a method of treatment of a disease associated with angiogenesis in a mammal, including a human, comprising administering to said mammal a sesquisuccinate or di-malonate salt of formula I in an amount effective to treat said disease. Such diseases include cancerous tumors, such as melanoma; ocular diseases such as age-related macular degeneration, presumed ocular histoplasmosis syndrome, and retinal neovascularization caused by proliferative diabetic nephropathy; rheumatoid arthritis; bone loss disorders such as osteoporosis, paget's disease, humoral hypercalcemia of malignancy, hypercalcemia resulting from metastasis of bone tumors, and osteoporosis induced by glucocorticoid therapy; restenosis of the coronary artery; and certain microbial infections, including those associated with microbial pathogens selected from the group consisting of adenovirus, hantavirus, borrelia burgdorferi, yersinia, bordetella pertussis, and group a streptococcus.

As used herein, "abnormal cell growth" means, unless otherwise indicated, cell growth that is not associated with normal regulatory mechanisms (e.g., loss of contact inhibition). This includes the following abnormal growths: (1) tumor cells (tumors) expressing an activated Ras oncogene; (2) tumor cells in which the Ras protein is activated by an oncogenic mutation of another gene; (3) benign and malignant cells in other proliferative diseases, in which disorganized Ras activation occurs; and (4) any tumor that is hyperplastic by farnesyl protein transferase.

The term "treating" as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progression of, or preventing the disease or disorder to which the term applies, or one or more symptoms of the disease or disorder. The term "treatment", as used herein, unless otherwise indicated, refers to the therapeutic effect of "treating" as defined above.

As used herein, the term "compound having reduced affinity for the erbB1 receptor" means, unless otherwise indicated, that the compound is an erbB2 inhibitor with a selectivity for the erbB2 receptor over the erbB1 receptor in the range of 50-1500, that is, the compound is 50-1500 fold more selective for the erbB2 receptor than the erbB1 receptor. In preferred embodiments, the erbB2 inhibitor has a selectivity over erbB1 for erbB2 over erbB1 in the range of 80-1000. In a still more preferred embodiment the erbB2 inhibitor has a selectivity over erbB1 for erbB2 over erbB1 in the range of between 90-500. In the most preferred embodiment, the erbB2 inhibitor has a selectivity over erbB1 over erbB2 in the range of 100-300. In the most preferred embodiment, the erbB2 inhibitor has a selectivity over erbB1 over erbB2 in the range of 110-200. The selectivity of erbB2 inhibitors over erbB1 inhibitors was measured using the whole cell (intact) assay described below.

Detailed description of the invention

The present invention relates to succinate and malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

In a preferred embodiment, the present invention relates to the sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

The present invention further relates to a process for the preparation of sesquisuccinate and di-malonate salts of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide. The salts of the present invention are useful in the treatment of hyperproliferative diseases, such as cancer, in mammals, especially humans, and to pharmaceutical compositions containing such compounds.

Salts of the compounds of formula I have been characterized using elemental analysis.

The in vitro activity of the compounds of formula I can be determined by the following method.

The in vitro activity of the compounds of formula I as erbB kinase inhibitors in whole cells can be determined by the following method. Cells, e.g., 3T3 cells transfected with human EGFR (Cohen et al J. virology 67: 5303, 1993) or chimeric EGFR/erbB2 kinase (EGFR extracellular/erbB 2 intracellular, Fazioli et al, mol. cell biol. 11: 2040, 1991) were plated in 96-well plates with 12,000 cells per well in 100. mu.l Dulbecco's Minimal Essential Medium (DMEM) containing 5% fetal bovine serum, 1% penicillin/streptomycin, 1% L-glutamine, cultured at 37 ℃ in 5% CO₂The following steps. Test compounds were solubilized in DMSO at a concentration of 10mM, and the final concentrations in the medium at the time of the assay were 0, 0.3. mu.M, 1. mu.M, 0.3. mu.M, 0.1. mu.M, and 10. mu.M. The cells were cultured at 37 ℃ for 2 hours. EGF (final 40ng/ml) was added to each well, cells were incubated at room temperature for 15 minutes, followed by aspiration of the medium, and then 100. mu.l/well of cold fixative (50% ethanol/50% acetone, containing)There were 200 micromoles of sodium orthovanadate). The plates were incubated at room temperature for 30 minutes, followed by washing with wash buffer (phosphate buffered saline containing 0.5% tween 20). Blocking buffer (phosphate buffered saline containing 3% bovine serum albumin, 0.05% tween 20, 200 μ M sodium orthovanadate, 100 μ l/well) was added followed by incubation at room temperature for 2 hours, followed by two washes with wash buffer. The PY54 monoclonal anti-phosphotyrosine antibody conjugated directly to horseradish peroxidase (50. mu.l/well with 1. mu.g/ml tissue buffer) or the conjugate from the tissue (1. mu.g/ml blocking buffer with 1mM phosphotyrosine to check for specificity) was added and the plates were incubated at room temperature for 2 hours. The plate wells were then washed 4 times with wash buffer. Addition of 50. mu.l of TMBMicrowell peroxidase substrate (Kirkegaard and Perry, Gaithersburg, Md.) per well generated a colorimetric signal, which was stopped by addition of 50. mu.l of 0.09M sulfuric acid per well. The absorbance at 450nM represents the phosphotyrosine content of the protein. Increased signal from EGF-treated cells compared to controls (no EGF treatment) represents the activity of EGFR or EGFR/chimera, respectively. The potency of the inhibitor was determined by measuring the concentration of compound required to inhibit phosphotyrosine in each cell line by 50%. By comparing IC against EGFR transfectants₅₀IC with chimeric transfectants against erbB2/EGFR₅₀The selectivity of the compounds over EGFR was determined for erbB 2. Thus, for example, IC on EGFR transfectants₅₀IC at 100nM for erbB2/EGFR chimeric transfectants₅₀Compounds at 10nM were considered to be 10-fold selective for erbB2 kinase.

Administration of the compounds of the present invention (hereinafter "active compounds") may be carried out by any method capable of delivering the compounds to the site of action. These methods include oral, intraduodenal, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical and rectal administration.

The amount of active compound administered will depend on the subject, the severity of the disease or condition, the rate of administration and the judgment of the attending physician. However, an effective dose is in the range of about 0.001 to about 100mg per kg body weight per day, preferably about 1 to about 35 mg/kg/day, administered in single or multiple doses. For a 70kg human, the dosage will reach about 0.05 to about 7 g/day, preferably about 0.2 to about 2.5 g/day. In some instances, dosage levels below the lower limit of the aforesaid range may be more suitable, while in other cases still larger doses may be employed without causing any harmful side effects, provided that such larger doses are first divided into several small doses for administration throughout the day.

The active compound may be used as a monotherapy or may include one or more other anti-tumour agents, for example selected from mitotic inhibitors, for example vinblastine; alkylating agents, such as cisplatin, carboplatin, and cyclophosphamide; antimetabolites, such as 5-fluorouracil, cytosine arabinoside and hydroxyurea, or one of the preferred antimetabolites, such as N- (5- [ N- (3, 4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino-, as disclosed, for example, in European patent application No. 239362]-2-thenoyl) -L-glutamic acid; a growth factor inhibitor; a cell cycle inhibitor; intercalating antibiotics, such as doxorubicin and bleomycin; enzymes, such as interferon; and anti-hormones, e.g. antiestrogens, e.g. Nolvadex^TM(tamoxifen), or for example antiandrogens, such as Casodex^TM(4 '-cyano-3- (4-fluorophenylsulfonyl) -2-hydroxy-2-methyl-3' - (trifluoromethyl) propionylaniline). Such combination therapy may be achieved by the simultaneous, sequential or separate administration of the individual therapeutic components.

The pharmaceutical composition may be, for example, a tablet, capsule, pill, powder, sustained release formulation, solution, suspension suitable for oral administration, a sterile solution, suspension or emulsion suitable for parenteral injection, an ointment or cream suitable for topical administration, or a suppository suitable for rectal administration. The pharmaceutical composition may be in unit dosage form suitable for single administration of a precise dose. The pharmaceutical compositions comprise conventional pharmaceutical carriers or excipients and the compounds according to the invention as active ingredients. In addition, it may include other medicinal or pharmaceutical agents, carriers, adjuvants, etc.

Exemplary parenteral dosage forms include solutions or suspensions of the active compounds in sterile aqueous solutions, such as aqueous propylene glycol or glucose solutions. Such dosage forms may be suitably buffered if desired.

Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents. The pharmaceutical compositions may contain additional ingredients, such as flavoring agents, binders, excipients, and the like, if desired. Thus for oral administration, tablets may be employed which contain various excipients, for example citric acid, together with various disintegrants for example starch, alginic acid and certain complex silicates, together with binding agents such as sucrose, gelatin and acacia. Additionally, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often useful for tableting purposes. Solid compositions of a similar type may also be employed in filled soft and hard gelatin capsules. Thus, preferred materials include lactose or milk sugar and macrogols. When aqueous suspensions or elixirs are desired for oral administration, the active compound may be combined with various sweetening or flavoring agents, coloring matter or dyes, and if desired, emulsifying or suspending agents, as well as diluents such as water, ethanol, propylene glycol, glycerin or combinations thereof.

Methods of preparing a variety of pharmaceutical compositions containing specific amounts of an active compound are known or will be apparent to those of skill in the art. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easter, Pa., 15th Edition (1975).

The following examples and preparations further illustrate and exemplify the compounds of the present invention and methods of making such compounds. It is understood that the scope of the present invention is by no means limited by the scope of the following examples and preparations. In the following examples, molecules with a single chiral center exist as a racemic mixture unless otherwise noted. Molecules with two or more chiral centers exist as racemic mixtures of diastereomers unless otherwise noted. The single enantiomers/diastereomers may be obtained by methods known to those skilled in the art.

If reference is made to HPLC chromatography in the preparations and examples below, the general conditions used are as follows, unless otherwise stated. The column used was ZORBAX^TMRXC18 column (manufactured by Hewlett Packard) 150mm long and 4.6mm inner diameter. Samples were tested on a Hewlett Packard-1100 system. The gradient solvent method used 100% ammonium acetate/acetic acid buffer (0.2M) to 100% acetonitrile over 10 minutes. The system then undergoes a wash cycle of first 1.5 minutes with 100% acetonitrile and then 3 minutes with 100% buffer solution. The flow rate for this phase was constant at 3 ml/min.

In the following examples and preparations, "Et" represents ethyl, "AC" represents acetyl, "Me" represents methyl, "ETOAC" or "ETOAc" represents ethyl acetate, "THF" represents tetrahydrofuran, and "Bu" represents butyl.

Example 1

Free base of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide

The free base of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide was prepared according to example 182 (LMRS: 470.1, HPLC RT: 5.05) using method G described in U.S. patent application Ser. No. 09/883,752, 6, 8, 2001, which is incorporated herein by reference in its entirety. Method G described in U.S. patent application Ser. No. 09/883,752 is as follows:

method G: synthesis of E-N- (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide (7)

E- (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino]-quinazolin-6-yl } -allyl) -carbamic acid tert-butyl ester: to a solution of 7.53ml of a 65% by weight solution of sodium bis (2-methoxyethoxy) aluminum hydride (Red-Al, 24.2mmol) in toluene in 90ml of tetrahydrofuran at 0 ℃ was added 5.0g of (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino]-quinaOxazolin-6-yl } -prop-2-ynyl) -carbamic acid tert-butyl ester as a solid. The reaction mixture was stirred at 0 ℃ for 2 hours, quenched with 10% aqueous potassium carbonate solution, and extracted with ethyl acetate. The organic layers were combined, dried and evaporated. The crude product was purified on 115g silica gel eluting with 80% ethyl acetate/hexanes to give 4.42g E- (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino]-quinazolin-6-yl } -allyl) -carbamic acid tert-butyl ester.¹H NMR(CDCl₃)：δ8.66(s，1)，8.24(m，1)，8.03(m，2)，7.77-7.65(m，3)，7.13(m，2)，6.97(d，J＝8.7Hz，1)，6.54(d，1)，6.35(m，1)，4.9(m，1)，3.90(m，2)，2.52(s，3)，1.46(s，9)。

E- [6- (3-amino-propenyl) -quinazolin-4-yl]- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenyl]-an amine. To 4.42g E- (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino]A solution of tert-butyl (4) -quinazolin-6-yl } -allyl) -carbamate in 21ml tetrahydrofuran was added 21ml2N hydrochloric acid. The mixture was heated at 60 ℃ for 3 hours, cooled to room temperature, and basified with 10% aqueous potassium carbonate. Methylene chloride was added to the aqueous mixture and a solid precipitated out. The solid was filtered and dried to give 2.98g E- [6- (3-amino-propenyl) -quinazolin-4-yl]- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenyl]-an amine.¹H NMR(d₆ DMSO)：δ8.62(s，1)，8.53(m，1)，8.26(m，2)，7.99(m，1)，7.89(m，1)，7.77(m，1)，7.30(m，3)，6.67(m，2)，344(m，2)，2.47(s，3)。

E-N- (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino]-quinazolin-6-yl } -allyl) -acetamide. A mixture of 14.4. mu.l (0.25mmol) of acetic acid and 40.3mg (0.33mmol) of dicyclohexylcarbodiimide in 2ml of dichloromethane is stirred for 10 minutes with 100.3mg of E- [6- (3-amino-propenyl) -quinazolin-4-yl]- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenyl]-amine treatment. The reaction mixture was stirred at room temperature overnight. The resulting precipitate was filtered and purified by silica gel chromatography eluting with 6-10% methanol in chloroform to give 106mg of the title compound; mp 254-.¹H NMR(d₆ DMSO)：δ9.88(s，1)，8.58(m，1)，8.20(m，3)，7.95(m，1)，7.83(m，1)，7.71(d，J＝8.7Hz，1)，7.24(m，2)，7.19(d，J＝8.7Hz，1)，6.61(d，J＝16.2Hz，1)，6.48(m，1)，3.90(m，2)。

Example 2

Sesquisuccinate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide

To a solution of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide in hot THF/acetone (5/100) was added two equivalents of succinic acid. As the solution cools, crystals slowly form. After stirring overnight, the crystals were filtered off and rinsed with acetone. The product was isolated as a white solid and confirmed by CHN analysis to be the sesquisuccinate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide. Calculated values: c61.29, H5.61, N10.83, experimental values: c is 61.04, H is 5.61, and N is 10.85.

Example 3

Di-malonate salt (di-malonate) of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide

To a solution of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide (1g) in hot acetone (100ml) was added two equivalents of malonic acid (443 mg). After 2 hours, crystals formed as the solution cooled, after stirring overnight, the crystals were filtered off and rinsed with acetone. Light yellow solid (1.36g, 94%) was confirmed by CHN analysis to be the di-malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide. Calculated values: c58.49, H5.21, N10.33, experimental values: c is 58.30, H is 5.12 and N is 10.33.

Claims (15)

1. The succinate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

2. The compound of claim 1, wherein the succinate salt is the sesquisuccinate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

3. A method of treating abnormal cell growth in a mammal comprising administering to said mammal an amount of the sesquisuccinate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide effective in treating abnormal cell growth.

4. The method according to claim 3, wherein the abnormal cell growth is cancer.

5. The method according to claim 4, wherein the cancer is selected from lung cancer, non-small cell lung (NSCL) cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head and neck, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, colon cancer, breast cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small bowel cancer, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urinary tract, cancer of the penis, cancer of the prostate, chronic and acute leukemias, lymphocytoma, cancer of the bladder, cancer of the kidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis, tumors of the Central Nervous System (CNS), colorectal cancer (CRC), primary CNS lymphoma, tumors of the spinal axis, brain stem glioma, pituitary adenoma, or a combination of one or more of the foregoing cancers.

6. A method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a compound of claim 1 effective to treat abnormal cell growth in combination with an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-hormones, and anti-androgens.

7. The method of claim 6 which comprises administering to said mammal an amount of a compound of claim 1 in combination with a cytotoxin effective to treat abnormal cell growth.

8. A method of treating abnormal cell growth in a mammal comprising administering to said mammal an amount of a compound of claim 1 effective to treat abnormal cell growth in combination with a compound selected from the group consisting of cyclophosphamide, 5-fluorouracil, floxuridine, gemcitabine, vinblastine, vincristine, daunorubicin, doxorubicin, epirubicin, tamoxifen, methylprednisolone, cisplatin, carboplatin, CPT-11, gemcitabine, paclitaxel, and docetaxel.

9. A pharmaceutical composition comprising an amount of a compound according to claim 1 effective to treat a hyperproliferative disorder in a mammal and a pharmaceutically acceptable carrier.

10. A malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

11. The compound of claim 10, wherein the malonate salt is a di-malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

12. A method of treating abnormal cell growth in a mammal comprising administering to said mammal an amount of a bis-malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide that is effective in treating abnormal cell growth.

13. A method of treating abnormal cell growth in a mammal which comprises administering to said mammal an amount of a compound of claim 12 effective to treat abnormal cell growth in combination with an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, anti-metabolites, intercalating antibiotics, growth factor inhibitors, radiation, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, antibodies, cytotoxins, anti-hormones, and anti-androgens.

14. A process for preparing a succinate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide comprising reacting E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide with succinic acid.

15. A process for preparing a malonate salt of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide comprising reacting E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide with malonic acid.