HK1082499A - Complexes of e-2-methoxy-n-(3-{4-[3-methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino]-quinazolin-6-yl}-allyl)-acetamide, their method of production, and use - Google Patents

Description

Complexes of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, methods of preparation and uses thereof

Background

The present invention relates to complexes 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 WO 01/98277 published 12/27 in 2001, which is incorporated herein by reference in its entirety. The prior application is designated as such. The free base of formula I is useful for the treatment of hyperproliferative disorders, such as cancer.

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, including sesquisuccinate and dimalonate salts, are disclosed in U.S. provisional patent application No.60/340885, filed 12/2001.

The invention further relates to specific complexes 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 comprising these complexes. The complexes of the invention are useful for treating hyperproliferative disorders, such as cancer, in mammals, particularly humans. The invention also relates to methods of administering these complexes to treat hyperproliferative disorders.

Summary of The Invention

The present invention relates to complexes 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.

Examples of such complexes include maleate (including dimaleate), hydrochloride (including monohydrochloride), succinate (including sesquisuccinate and monosuccinate), malonate (including dimalonate), phosphate (including monophosphate), fumarate (including monofumarate), hemiedisylate (hemiedisylate), tartrate (including racemate and optically active forms), camsylate (including racemate and optically active forms), benzenesulfonate, ethanesulfonate, nitrate, and citraconate (including dicitraconate) complexes of formula I.

The invention also relates to a complex formed from E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide and an acid or a reactive equivalent of said acid, wherein said acid is at least one selected from the group consisting of maleic acid, hydrochloric acid and phosphoric acid.

The present invention also relates to a method of inhibiting abnormal cell growth in a mammal comprising administering to said mammal an amount of a complex effective to inhibit abnormal cell growth as described above.

The invention also relates to a method of treating a mammal suffering from a disorder characterized by overexpression of erbB2, such as cancer, comprising administering to the mammal an amount of the above complex effective to treat the disorder.

The invention also relates to a method of inducing cell death comprising exposing cells which overexpress erbB2 to an effective amount of the above complex.

The invention also relates to a pharmaceutical composition comprising an amount of the above complex effective to treat hyperproliferative disorders in a mammal, and a pharmaceutically acceptable carrier.

Brief Description of Drawings

FIG. 1 is an X-ray powder diffraction pattern of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide monohydrochloride prepared and isolated according to example 5.

FIG. 2 is an X-ray powder diffraction pattern of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide dimaleate prepared and isolated according to example 6.

FIG. 3 is an X-ray powder diffraction pattern of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide monophosphate (monohydrate) as described in example 7.

Detailed Description

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

formula I.

Examples of such complexes include the maleate (including dimaleate), hydrochloride (including monohydrochloride), succinate (including sesquisuccinate and monosuccinate), malonate (including dimalonate), phosphate (including monophosphate), fumarate (including monofumarate), hemiedisylate, tartrate (including racemate and optically active forms), camsylate (including racemate and optically active forms), benzenesulfonate, ethanesulfonate, nitrate, and citraconate (including dicitraconate) complexes of formula I.

In a preferred embodiment the invention relates to hydrochloride, maleate and phosphate complexes of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide.

In a particularly preferred embodiment, the hydrochloride complex is a monohydrochloride complex, the maleate complex is a dimaleate complex and the phosphate complex is a monophosphate complex.

In a preferred embodiment, the dimaleate salt, the monohydrochloride salt, and the monophosphate complex are substantially salts.

In one embodiment, the presently disclosed monohydrochloride, monophosphate, and dimaleate complexes of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide are amorphous, and in one embodiment (preferred) are crystalline, i.e., substantially free of amorphous material (i.e., at least 90% crystalline, and in one embodiment, at least 95% crystalline, and in one embodiment at least 99% crystalline). Such crystalline materials can provide more reproducible dosing results. They have optimum aqueous solubility, chemical and physical stability and bioavailability for pharmaceutical compositions. They generally have relatively high solubility and bioavailability compared to the E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide starting material from which they are prepared. The stability of these materials may also reduce potential problems associated with weight changes of the active ingredient in the preparation of capsules or tablets.

In one embodiment, the hydrochloride, dimaleate and monophosphate salts are crystalline materials that exhibit X-ray powder diffraction patterns having characteristic peaks expressed in degrees (2 θ) and Relative Intensities (RI) as disclosed in examples 3, 4 and 5, respectively.

The dimaleate, monophosphate, and monohydrochloride complexes of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide are chemically stable and non-hygroscopic, which may reduce potential problems associated with weight changes of the active ingredient in the preparation of capsules or tablets.

The invention also relates to a complex formed from E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide and an acid or a reactive equivalent of said acid, wherein said acid is at least one selected from the group consisting of maleic acid, hydrochloric acid and phosphoric acid.

In one embodiment, where the acid is maleic acid, the complex is E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide maleate and preferably E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide dimaleate.

In one embodiment, where the acid is hydrochloric acid, the complex is E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide hydrochloride and preferably E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide monohydrochloride.

In one embodiment, where the acid is phosphoric acid, the complex is E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide phosphate and preferably E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide monophosphate.

The present invention also relates to a method of inhibiting abnormal cell growth in a mammal comprising administering to said mammal an amount of the above-described complex of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, which is effective in inhibiting abnormal cell growth.

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

In one embodiment herein, the cancer is selected from lung cancer, non-small cell lung (NSCL) cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or 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 or acute leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, cancer of the renal cell, cancer of the renal pelvis, tumor of the Central Nervous System (CNS), colorectal cancer (CRC), primary CNS lymphoma, spinal cord tumor, brain stem glioma, 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 hypertrophy or restinosis.

In one embodiment of the invention, the cancer is selected from breast cancer, colon cancer, ovarian cancer, non-small cell lung (NSCL) cancer, colorectal cancer (CRC), prostate cancer, bladder cancer, kidney cancer, stomach cancer, endometrial cancer, head and neck cancer, and esophageal cancer.

In a further 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 invention is directed to a method of inhibiting abnormal cell growth in a mammal comprising administering to the mammal an amount of a complex 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 inhibiting abnormal cell growth, binding an antineoplastic 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-hormonal agents, and anti-androgens.

In a preferred embodiment, the complex binds to a cytotoxin.

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

The present invention further relates to a method of inhibiting abnormal cell growth in a mammal comprising administering to said mammal an amount of a complex 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 inhibiting 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 above compound is selected from tamoxifen, cisplatin, carboplatin, paclitaxel and docetaxel.

The present invention further relates to a pharmaceutical composition for inhibiting abnormal cell growth in a mammal comprising an amount of a complex 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 inhibiting abnormal cell growth, and a pharmaceutically acceptable carrier.

In one embodiment, the pharmaceutical composition further comprises an antineoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.

The present invention also relates to a method of treating a mammal having a disease characterized by overexpression of erbB2 comprising administering to said mammal a complex of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide in an amount effective to treat said disease characterized by overexpression of erbB 2.

In a preferred embodiment, the disease is cancer.

The invention also relates to a method of inducing cell death comprising exposing cells over-expressing erbB2 to an effective amount of a complex of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide. In one embodiment, the cell is a mammalian, preferably human, cancer cell.

The present invention relates to a method of inducing cell death comprising exposing cells over-expressing erbB2 to an effective amount of a complex of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, and the method further comprises exposing the cells to 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 comprising administering to the human a therapeutically effective amount of a complex of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, which has reduced affinity for the erbB1 receptor. 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 present invention also relates to a method of treating a disease associated with angiogenesis in a mammal, including a human, comprising administering to said mammal a complex of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, or a solvate or prodrug thereof, that is effective in treating said disease. The disease includes cancerous tumors, such as melanoma; ophthalmic diseases, age-related macular degeneration, presumed ocular histoplasmosis syndrome, and retinal neovascularization due to proliferative diabetic retinopathy; rheumatoid arthritis; bone loss diseases such as osteoporosis, Paget's disease, humoral hypercalcemia of malignancy, hypercalcemia of tumor metastasis to bone, and osteoporosis caused by glucocorticoid therapy; restenosis of the coronary arteries; and include certain microbial infections associated with microbial pathogens selected from the group consisting of adenovirus, hantavirus, borrelia burgdorferi, yersinia, bordetella pertussis, and group a streptococcus.

As used herein, unless otherwise specified, "complex" refers to a defined stoichiometric acid-base pair and includes ionized, non-ionized, and/or partially charged bases and acids, where the degree of proton transfer from the acid (proton donor) to the base (proton acceptor) can vary proportionally from none, partial, to all. All complexes may be defined with the suffix "ate" or "ide" to indicate the complex of the particular acid whose name ends with "ic". For example, a complex of a basic compound with succinic acid in which the molar ratio of succinic acid to the basic compound is 1.5 is named as "sesquisuccinate" of the basic compound. One of ordinary skill in the art will appreciate that the above definition of "complex" includes salts in which the degree of proton transfer from the acid to the base is substantially proportional to the total (i.e., complete proton transfer).

As used herein, "substantially salt" refers to a compound in which the degree of proton transfer from the acid to the base is at least about 90%, and in one embodiment, at least about 95%, and in one embodiment, at least about 99%.

As used herein, "reactive equivalent of a substance" refers to any compound or chemical composition, other than the substance itself, that can react under reaction conditions as the substance itself does. Thus, reactive equivalents of carboxylic acids shall include acid-generating derivatives such as anhydrides, acyl halides and mixtures thereof unless otherwise specifically indicated. One of ordinary skill in the art will recognize that the phrase "synthon" is synonymous with "reaction equivalent".

As used herein, "abnormal cell growth," unless otherwise indicated, refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes abnormal growth: (1) tumor cells (tumors) expressing the activated Ras oncogene; (2) tumor cells in which Ras protein is activated as a result of oncogene mutation in another gene; (3) benign and malignant cells of other proliferative diseases, in which abnormal Ras activation occurs; and (4) any tumor that proliferates due to farnesyl protein transferase.

The term "treating", as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progression of, or preventing the disease or condition for which the term is applied, or one or more symptoms of the disease or condition. The term "treatment", as used herein, unless otherwise indicated, refers to the act of treating by "treating" as defined above.

The term "a compound which has reduced affinity for the erbB1 receptor", as used herein, unless otherwise indicated, means a compound wherein the compound is an erbB2 inhibitor and is selective for the erbB2 receptor over the erbB1 receptor by between 50-1500, i.e. the compound is selective for the erbB2 receptor over the erbB1 receptor by between 50-1500 times. In a preferred embodiment the erbB2 inhibitor is more selective for erbB2 than erbB 160-1200. In a more preferred embodiment the erbB2 inhibitor is selective for erbB2 over erbB 180-1000. In an even more preferred embodiment the erbB2 inhibitor is more selective for erbB2 than erbB 190-500. In a most preferred embodiment the erbB2 inhibitor is selective for erbB2 over erbB 1100-300. In this most preferred embodiment the erbB2 inhibitor is selective for erbB2 over erbB 1110-200. The selectivity of the erbB2 inhibitor over the erbB1 inhibitor was determined using a whole cell (intact) assay as described below.

For all purposes, each document is incorporated herein by reference in its entirety. Except in the examples, or where otherwise expressly indicated, all numerical quantities in this description specifying amounts of material, degree of crystallinity, degree of proton transfer from acid to base as described in the foregoing description of "complexes", reaction and process conditions (e.g., temperature, time, pressure), and the like, are to be understood as modified by the word "about".

The in vitro activity of the complex of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide can be determined by the following procedure.

E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] as erbB kinase inhibitors in intact cells]The in vitro activity of the complexes of-quinazolin-6-yl } -allyl) -acetamide can be determined by the following procedure. Cells, such as 3T3 cells transfected with human EGFR (Cohen et al J. virology 67: 5303, 1993) or with chimeric EGFR/erbB2 kinase (EGFR extracellular/erbB 2 intracellular, Fazioli et al mol.cell.biol.11: 2040, 1991) were placed in 96-well plates in 12,000 cells per well in 100. mu.l of medium (Dulbecco's minimal essential Medium (DMEM) containing 5% fetal bovine serum, 1% penicillin/streptomycin, 1% L-glutamine) and at 37 ℃, 5% CO₂Culturing in medium. Test compounds were dissolved in DMSO at a concentration of 10mM, and the final concentrations in the medium were 0, 0.3. mu.M, 1. mu.M, 0.3. mu.M, 0.1. mu.M, and 10. mu.M for the assay. Cells were cultured at 37 ℃ for 2 hours. EGF (40ng/ml final) was added to each well and the cells were incubated at room temperature for 15 minutes, followed by aspiration of the medium and then addition of 100. mu.l/well of cold fixative (50% ethanol/50% acetone containing 200 micromoles sodium orthovanadate). The plates were incubated at room temperature for 30 minutes and then washed with wash buffer (0.5% Tween 20 in phosphate buffered saline). Blocking buffer (3% bovine serum albumin, 0.05% Tween 20, 200. mu.M sodium orthovanadate in phosphate buffered saline, 100. mu.l/well) was added followed by incubation at room temperature for 2 hours, followed by washing twice with washing buffer. Direct binding (50. mu.l/well, 1. mu.g/ml in blocking buffer) or coating binding (in block) was addedBlocking buffer containing 1mM phosphotyrosine at 1. mu.g/ml to check specificity) PY54 monoclonal anti-phosphotyrosine antibody to horseradish peroxidase, and the plates were incubated at room temperature for 2 hours. The plate wells were then washed 4 times with wash buffer. The colorimetric signal was enhanced by adding 50. mu.l of TMB microwell peroxidase substrate (Kirkegaard and Perry, Gaithersburg, Md.) per well and stopped by adding 50. mu.l of 0.09M sulfuric acid per well. Absorbance at 450nM indicates phosphotyrosine content of the protein. Increased signal in EGF-treated cells over controls (non-EGF treatment) indicates the activity of EGFR or EGFR/chimera, respectively. Inhibition of phosphotyrosine increase by 50% (IC) in each cell line was determined₅₀) The concentration of the compound required to determine the activity of the inhibitor. IC by comparison of EGFR transfectants to erbB2/EGFR chimera transfectants₅₀To determine the selectivity of the compound for erbB2 for EGFR. Thus, for example, having IC for EGFR transfectants₅₀IC at 100nM and transfectants to the erbB2/EGFR chimera₅₀Compounds at 10nM are believed to be 10-fold more selective for erbB2 kinase.

Administration of the compounds of the present invention (hereinafter "active compounds") is effected by any method that is capable of delivering the compound to the reaction site. These methods include oral routes, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical and rectal administration.

The amount of active compound administered will depend on the individual being treated, the severity of the disease or condition, the rate of administration and the judgment of the prescribing physician. However, an effective dose is from about 0.001 to about 100mg per kg body weight per day, preferably from about 1 to about 35 mg/kg/day, administered alone or in divided doses. For a 70kg human, this possible dosage is 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 instances, larger doses may be employed without causing any harmful side effects, provided that the larger dose is first divided into smaller doses for administration throughout the day.

The activation is carried outThe compounds may be used as monotherapy or may include one or more other anti-tumour agents, such as those selected from: for example mitotic inhibitors such as vinblastine; alkylating agents, such as cisplatin, carboplatin, and cyclophosphamide; antimetabolites, such as 5-fluorouracil, cytarabine and hydroxyurea, or a preferred antimetabolite as disclosed in European patent application No.239362, such asN-(5-[ N- (3, 4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino radical]-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-hormonal agents, e.g. anti-estrogens, e.g. Nolvadex^TM(tamoxifen) or, e.g., antiandrogenic agents, 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 ingredients.

The pharmaceutical compositions may be administered orally, for example in a suitable form such as tablets, capsules, pills, powders, sustained release formulations, solutions, suspensions, parenterally in the form of sterile solutions, suspensions or emulsions, topically in the form of ointments or creams or rectally in the form of suppositories. The pharmaceutical composition may be in unit dosage form suitable for single administration at a precise dosage. Pharmaceutical compositions will include conventional pharmaceutical carriers or excipients and the compounds of the invention as active ingredients. In addition, it may include other medical or pharmaceutical agents, carriers, adjuvants, and the like.

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

Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents. If desired, the pharmaceutical composition may contain additional ingredients such as flavoring agents, binders, excipients, and the like. Thus, for oral administration, tablets containing various excipients, such as citric acid, may be employed with various dispersing agents, such as starch, alginic acid and certain complex silicates, and with binding agents, such as sucrose, gelatin and acacia. Additionally, lubricants, such as magnesium stearate, sodium lauryl sulfate and talc are commonly used to prepare tablets. Solid compositions of a similar type may also be employed in soft and hard filled gelatin capsules. Thus, preferred materials include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration of the active compounds therein, various sweetening or flavoring agents, coloring matter or pigments and, if desired, emulsifying agents or suspending agents may be combined for use with diluents such as water, ethanol, propylene glycol, glycerin or combinations thereof.

Methods for preparing various pharmaceutical compositions containing specific amounts of active compound are known or will be apparent to those skilled in the art. See, for exampleRemington′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 the compounds. It will be understood that the scope of the present invention is not limited by the scope of any of the following examples and preparations. In the following examples, molecules having one chiral center, unless otherwise noted, are racemic mixtures. Those molecules with two or more chiral centers, unless otherwise noted, are racemic mixtures of diastereomers. The individual enantiomers/diastereomers may be obtained by methods known to those skilled in the art.

In the HPLC chromatographs mentioned in the preparations and examples below, the conventional conditions used are as follows, unless otherwise stated. The column used was ZORBAX 150mm long and 4.6mm inner diameter^TMRXC18 column (manufactured by Hewlett Packard). The samples were run on a Hewlett Packard-1100 system. A gradient solvent method was used, eluting with 100% ammonium acetate/acetic acid buffer (0.2M) to 100% acetonitrile for 10 minutes. Then, the reaction was carried out with 100% acetonitrileThe rinse cycle was 1.5 minutes, and then rinsed with 100% buffer solution for 3 minutes. The flow rate at this stage was a constant flow of 3 ml/min.

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

FIGS. 1-3 Spectroscopy Using Bruker with copper radiation, fixed slits (1.0, 0.6mm), and Kevex solid State Detector¹D5000 diffractometer recordings. Data were collected at 3.0-40.0 degrees 2 theta with step size 0.04 degrees and step time 1.0 second.

The test conditions for powder X-ray diffraction were: a Cu target; wavelength 1: 1.54056 angstroms; wavelength 2: 1.54439 angstroms (relative intensity: 0.500); range # 1-coupling: 3.000 to 40.000; step length: 0.040; when the steps are carried out: 1.00; smooth width: 0.300 parts by weight; and a threshold value: 1.0.

for single crystal X-ray analysis, data were collected using a Bruker CCD diffractometer. Cu target: a wavelength of 1.54178 angstroms; room temperature;

the following details relate to data analysis: the atomic scattering factor is obtained from the International Table of X-ray crystallography (Vol.IV, pp.55, 99, 149 Birmingham: Kynoch Press, 1974). All crystallographic calculations were performed by means of the SHELXTL system g.m. sheldrick, SHELXTL, User Manual, Nicholet Instrument co., 1981). Test structures were obtained by the direct method.

PXRD pattern calculations based on single crystal data: to compare the results between single crystals and powder samples, powder X-ray patterns based on single crystal structure data can be calculated. Can be calculated using the SHELXTL additive computer program, see the reference manual of Siemens Analytical X-ray Instrument, Chapter 10, p.179-181, 1990. The single crystal structure data provides the unit cell size, space group and atomic position of the crystal form. These parameters serve as the basis for calculating the correct powder mode for the crystal form. Comparing the calculated PXRD pattern with the experimental pattern will further confirm whether the powder sample conforms to the specified single crystal structure. This procedure has been used for the crystalline forms of azithromycin A, D, F, G and J. The results are shown in the superimposed powder X-ray diffraction pattern of the lower pattern calculated from the single crystal data and the higher pattern as a representative experimental pattern. The match between the two patterns shows the consistency between the powder sample and the corresponding single crystal structure.

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 procedure G disclosed in PCT publication WO 01/98277, the disclosure of which is incorporated herein by reference in its entirety. Procedure G in WO 01/98277 is shown below.

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: 5.0g of solid (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino) -at 0 ℃ are added]-quinazolin-6-yl } -prop-2-ynyl) -carbamic acid tert-butyl ester was added to a solution of 7.53ml of bis (2-methoxyethoxy) aluminum hydride sodium salt (Red-Al, 24.2mmol) in 90ml of tetrahydrofuran in 65% by weight toluene. The reaction was stirred at 0 ℃ for 2 hours, quenched with 10% aqueous potassium carbonate and extracted with ethyl acetate. The combined organics were dried and evaporated. The crude extract 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. 21ml of 2N hydrochloric acid was added to a solution of 4.42g E- (3- {4- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] in 21ml of tetrahydrofuran]-quinazolin-6-yl } -allyl) -carbamic acid tert-butyl ester. The mixture was heated at 60 ℃ for 3 hours, cooled to room temperature and basified with 10% aqueous potassium carbonate. Dichloromethane was added to the aqueous mixture and a solid precipitated. 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(d₆DMSO)：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)，3.44(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 and 100.3mg of E- [6- (3-amino-propenyl) -quinazolin-4-yl]- [ 3-chloro-4- (6-methyl-pyridin-3-yloxy) -phenyl]-amine treatment. The reaction was stirred at room temperature overnight. The precipitate formed was filtered and chromatographed on a column of silica gel, eluting with 6-10% methanol/chloroform to give 106mg of the title compound; m.p.254-256 deg.c;¹H-NMR(d₆DMSO)：δ9.88(s，1)，8.58(s，1)，8.48(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

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

The following procedure for the preparation of the free base of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide is disclosed in U.S. provisional application 60/334647, filed on 30/11/2001:

synthesis of 6-iodo- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazoline:

the mechanical stirrer is put into a three-neck round-bottom flask and is kept in N₂In (1). The flask was charged with 6-iodo-4-chloroquinazoline (10.0g, 34.43mol) and anhydrous THF (35 ml). Then 3-methyl-4- (6-methyl-pyridin-3-yloxy) -aniline (7.38g, 34.43mmol) and anhydrous THF (45ml) were added and the yellow suspension was heated to reflux. After 15 minutes, most of the reaction dissolved into solution and a fine yellow suspension was obtained. After 25 minutes, the internal temperature of the reaction mixture was 56 ℃ and the desired product began to precipitate. Heating was continued for an additional 2 hours and the reaction mixture was cooled to room temperature and held in an oil bath. The yellow crystals were collected by filtration, washed with cold (0 ℃) THF (1X 10ml) and dried at 50 ℃ with p < 200 mbar. The title compound was obtained as pale yellow crystals (15.75g, 98%). R_f＝0.45(EtOAc/MeOH＝9/1)。¹H-NMR(CDCl₃，300MHz)：δ＝11.40(br，s，1H，NH)，9.29(d，J＝Hz，1H， H-2)，8.91(s，1H， H-2″)，8.36-8.32(m，2H， H-7， H-8)，7.74-7.73(m，2H， H-4″， H-5)，7.62(dd，J₁＝8.7Hz，J₂＝2.6Hz，1H， H-5″)7.49-7.46(m，2H，H-6′， H-5)，7.06(d，J＝8.7Hz，1H， H-2′)，2.54(s，3H，CH₃)，2.26(s，3H，CH₃)。¹³C NMR(CDCl₃+D₆-DMSO，75MHz)：δ＝159.51，153.63，153.17，152.82，152.70，145.26，141.37，138.01，134.75，134.65，131.05，129.10，128.74，126.77，124.86，124.43，120.41，116.98，94.89，23.54，17.67。

The title compound is shown below as RP-HPT under LC condition_R(min.) 12.13: symmetrical shielding RP18, 75 × 4.6 mm; the flow rate is 1.0 ml/min; 205/210/220/245 nm; the temperature is 25 ℃; sample introduction volume: ACN/H₂10 μ L of approximately 0.5% solution in O9/1; eluent: b: ACN, C: at H₂0.01mmol of NH in O₄OAc, pH 6.0; and gradient: and (3) 0 minute: b is 30%, C is 70%; and 20 minutes: b is 85% and C is 15%.

Synthesis of 2-methoxy-acetic acid propargylamide:

in N₂In anhydrous CH₂Cl₂A solution of methoxyacetyl chloride (12.5ml, 0.137mol, 1.2 eq.) in (45ml) was cooled to-40 ℃. Maintaining the temperature below-25 deg.C, adding anhydrous CH₂Cl₂A solution of propargylamine (7.98ml, 0.125mol, 1.0 eq.) in (40ml) was over 45 minutes. After 15 minutes, triethylamine (17.4ml, 0.125mol, 1.0 eq) was added over 45 minutes, maintaining below-25 ℃. The reaction mixture was warmed to room temperature. TLC after 3 hours showed complete conversion. By H₂The reaction mixture was quenched with O (50ml) and the organic phase was washed with half-saturated NaCl solution, filtered through cotton wool and concentrated at a pressure of more than 650mbar and 40 ℃. The crude compound was purified by short-neck distillation (boiling point 49 ℃ C. and pressure 0.09 mbar). Crystallization was carried out by a conventional method to give the title compound as a colorless liquid (7.84g, 50%).

R_f0.36. (heptane/ethyl acetate 7/3).

¹H NMR(CDCl₃，300MHz)：δ＝6.72(br，s，1H，N-H)，4.09(dd，J₁＝5.5Hz，J₂＝2.6Hz，2H，CH₂-NH)，3.92(s，2H，CH₂-OMe)，3.43(s，3H，OCH₃) 2.24.(t, J ═ 2.6Hz, 1H, alkyne CH)

¹³C-NMR：(CDCl₃，75MHz)δ＝169.14(C＝O)，79.11( C-2′)，71.63( C-2)，71.41( C-3′)，59.04(OCH₃)，28.26.( C-1′)。

T was measured by gas chromatography under the conditions shown in the following table_RThe (minute) was 6.42.

Chromatographic column	DB-5(30m 0.32mm, 0.25 μm film thickness)
		Sample injector	Splitting at 250 deg.C
Split ratio	60.243：1
		Diverting fluid	108.3ml/min, carrier gas type: hydrogen gas
Column oven	60℃，1min，10℃/min，290℃，10min
		Sample introduction temperature	250℃
Detector (FID)	The detection temperature is 250 DEG C
		Detecting fluid	H2: 40.0ml/min, air: 450ml/min
Make-up fluid	N2：45.0ml/min

Preparation of 6- (N-methoxyacetyl-3-amino-propen-1-yl) -4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazoline, which is E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide

Using a Suzuki coupling reaction:

2-methyl-2-butene (0.59ml, 5.60mmol, 2.8 equiv.) was added to cold (0-5 ℃ C.) BH over 1 hour₃THF complex solution (1.0M solution, 3.0ml, 3.0mmol, 1.5 equiv.) in N₂And (4) maintaining.

The reaction mixture was stirred at this temperature for 30 minutes, followed by the addition of 2-methoxy-acetic acid propargylamide (255mg, 2mmol, 1.0 equiv.) dissolved in anhydrous THF (1ml) over 15 minutes. The ice bath was removed and the reaction mixture was warmed to room temperature for 20 minutes or more. The reaction mixture was then heated at 35 ℃ for 1 hour. Dissolving in degassing H₂K in O (1.2ml)₂CO₃(0.55g, 4mmol, 2.0 equiv.) was added to the reaction mixture over 30 minutes. Gas evolution was observed at the first half of the addition and stopped at further additions. Adding 6-iodo- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino group to the three parts]Quinazoline (1.41g, 3mmol, 1.5 equiv.) gave a yellow suspension. Mixing PPh₃(21mg, 0.08mmol, 4 mol%) and Pd (OAc)₂(4.5mg, 0.02mmol, 1 mol%) was added to each portion and the reaction mixture was heated to reflux (65-68 ℃). After about 30 minutes, a yellow solution was obtained and the reaction was checked by HPLC detection. After 18 h, the reaction mixture was cooled to room temperature, followed by addition of half-saturated NaCl solution (10ml) and EtOAc (10 ml). Separating the organic phase from H₂O (5ml) wash and at 50 ℃ and less than 200mbarConcentrated under pressure of (1). Filtering and purifying by a plug to obtain SiO₂EtOAc/MeOH 9/1. The title compound was obtained as pale yellow crystals (0.55g, 59%). R_f＝0.16(EtOAc/MeOH＝9/1)。¹H-NMR：(CDCl₃，250MHz)δ＝8.71(s，1H， H-2)，8.25(d，J＝1.7Hz，1H， H-8)，7.90(s，1H， H-7)，7.82(s，1H，NH)，7.79(s，1H， H-5)，7.66(d，J＝2.5Hz，1H， H-4″)，7.54(dd，J₁＝8.7Hz，J₂＝2.6Hz，1H， H-5″)，7.15-7.07(m，2H， H-5′， H-6′)，6.91(d，J＝8.7Hz，1H， H-2′)，6.83(bt，1H，NH)，6.6(d，J＝15.9Hz，1H， H-9), 6.34 and 6.29(dt, J)₁＝15.9Hz，J₂＝6.1Hz，1H， H-10)，4.14(dt，J＝6.1Hz，2H，CH₂OMe))，3.97(s，2H，CH₂NH)，3.45(s，3H，OCH₃)，2.53(s，3H，CH₃)，2.29(s，3H，CH₃)。¹³C-NMR(CDCl₃，75MHz)：δ＝169.79(C＝O)，157.90，154.93，152.367，152.23，150.90，149.74，139.34，134.73，134.63，131.16，130.77，130.36，128.85，129.98，125.47，124.66，123.65，121.32，119.51，119.13，115.39，71.96，59.26，40.84，23.57，16.41。

The t of the title compound was determined by reverse phase high performance liquid chromatography under the conditions shown in the following table_R(min) is 6.02

Symmetrical shield RP18	75×4.6mm
		Flow rate of flow	1.0ml/min
Wavelength of light	205/210/220/245nm
		Temperature of	25℃
Sample introduction volume	At ACN/H2About 0.5% solution in O9/1 10. mu.L
		Eluent B	ACN
Eluent C	At H20.01mmol NH in O4OAc，pH＝6.0
		Gradient 0min	B＝30％，C＝70％
Gradient for 20min	B＝85％，C＝15％

Example 3

The 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 form of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide may also be prepared by neutralization of the corresponding dimesylate salt.

The bis-mesylate salt was prepared as follows:

67.33 g were mixed in 400ml EtOH and 100ml CH at room temperature₂Cl₂In (b) E-2-methoxy-N- (3- {4- [ 3-methyl-4-, (6-methyl-pyridin-3-yloxy) -phenylamino]The free base of (E) -quinazolin-6-yl } -allyl) -acetamide (prepared as described in example 1 above) was added dropwise to 19.17ml (2.05) of equivs methanesulfonic acid (CH) in 100ml acetonitrile₃SO₃H) And (3) solution. The mixture was homogenized for 15 minutes at room temperature, then dichloromethane (. about.100 ml) was removed. An additional 600ml of acetonitrile was added to complete crystallization and the mixture was homogenized for 2 hours. The crystals were filtered under nitrogen and washed with 100ml acetonitrile. The yield of the bis (mesylate) (94.48 g) was 99%.

The bis-mesylate salt (90g) prepared according to the method of the preceding paragraph was dissolved in water (. about.550 ml). Chloroform (. about.500 ml) was added to the solution followed by 1N NaOH until a white suspension/precipitate (pH. about.13-14) was observed. Chloroform was added prior to NaOH to reduce stickiness due to precipitate formation. The mixture was transferred to a separatory funnel (2L) and the free base was extracted with three portions of chloroform (. about.300 ml). The extracts were combined (. about.1.3L), washed with water (. about.500 ml), dried over anhydrous magnesium sulfate and filtered. The chloroform filtrate was concentrated in vacuo to give a yellow amorphous solid/oil. This material was repeatedly homogenized in ethyl acetate overnight to yield a white solid. This material was then filtered, washed with cold ethyl acetate, and then dried in a vacuum oven at 45 ℃ to yield a white crystalline solid (-59 g). The free base was identified by Polarized Light Microscopy (PLM), powder X-ray diffraction (PXRD), and differential thermal scanning (DSC). It is needle-shaped and DSC shows three endothermic points (DSC melting point: 125 ℃, 160 ℃ and 167 ℃).

Example 4

Synthesis of E-2-methoxy-N- (3- (4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl) -allyl) -acetamide monohydrochloride:

a solution of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide in isopropanol was prepared by the procedure of example 1, 2 or 3 by dissolving 500mg of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide in 50ml of isopropanol with stirring. The solution was heated to 75 ℃. Then, concentrated hydrochloric acid (1.1 eq; 115mg) was diluted with 6ml of isopropanol. The diluted HCl solution was added dropwise with stirring to the hot free base solution. After complete addition, the solution was not heated and cooled to room temperature for more than about 3 hours, resulting in a fine crystalline precipitate. The dark yellow slurry was stirred for one day and filtered. The yellow fine powder was collected by vacuum filtration and dried in vacuo. The yield was about 79%.

The hydrochloride salt was determined to be the anhydrous monohydrochloride salt by combustion analysis. The melting endotherm of the compound at 222 ℃ was determined by DSC at a temperature rise rate of 5 ℃/min. Its PXRD pattern is shown in FIG. 1. Characteristic X-ray powder diffraction peaks (2 θ (± 0.1 °) [% relative intensity ]): 4.6[100], 9.3[20.9], 11.4[10.6], 15.6[3.4], 16.4[2.8], 17.1[11.8], 18.4[34.8], 18.8[5.9], 20.1[3.8], 20.4[8.6], 22.6[8.2], 23.0[5.1], 24.0[3.3], 25.4[2.7], 25.8[3.7], 27.5[10.7], and 28.3[3.2 ].

Example 5

Synthesis of E-2-methoxy-N- (3-4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl) -allyl) -acetamide dimaleate:

maleic acid solution was used at 7: 3(v/v) CHCl₃2.2 equivalents of maleic acid in EtOH. E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino]-quinazolin-6-yl } -allyl) -acetamide (prepared as described in example 1, 2 or 3 above) was dissolved in 70: 30CHCl₃In EtOH (v/v) and the maleic acid solution was added dropwise with stirring. After about 2 days, a white crystalline powder precipitated.

With polarized light microscopy, the dimaleate crystals are needle crystals with strong birefringence. In a hot state Polarized Light Microscope (PLM), the crystals melt/decompose at 170 ℃. The DSC thermal curve shows an endothermic curve followed by an exothermic curve at 170 ℃. Endothermic and exothermic curves corresponding to melting/decomposition were observed with the hot PLM. Moisture absorption: at 90% relative humidity, 0.6% by weight. PXRD is shown in FIG. 2. Characteristic X-ray powder diffraction peaks (2 θ (± 0.1 °), [% relative intensity ]): 4.6[20.4], 6.0[41.9], 7.2[13.1], 9.4[33], 9.7[32], 11.2[27.7], 12.0[5.2], 14.1[20], 14.2[53], 15.5[63.7], 15.7[51.2], 18.4[55], 18.7[93.4], 19.3[5], 19.6[21.9], 20.2[22.9], 20.4[16.2], 20.8[15.5], 21.2[37.6], 22.4[22.7], 22.8[68.7], 23.2[49.2], 23.4[62.5], 23.8[18.8], 24.5[8.7], 24.8[34.3], 25.2[100], 25.7[18.4], 26.4[11.5], 23.9.5 ], 23.8[18.8 [ 8], 24.7 [ 8[ 8.7], 24.9.9.9.9, 29.9[ 9.9, 29.9.9, 9, 29.9, 9, 29.6[ 6[ 6.6.6 [30 [ 6.7, 9, 29.7[ 6[ 6.7 ] and 29.7. Calculated X-ray diffraction peaks (from single crystals) (2 θ (± 0.1 °), [% relative intensity ]): 4.7[21], 6.0[34.5], 7.2[18.3], 9.5[32.3], 9.7[25.9], 11.3[32], 12.1[1.7], 14.0[20.3], 14.2[37.8], 15.6[37.5], 15.8[42.1], 18.4[59.7], 18.8[100], 19.3[15.9], 19.7[22.9], 20.2[22.9], 20.5[16.5], 20.8[186], 21.3[58.8], 22.4[29.5], 22.8[75.9], 23.3[48.3], 23.5[55.7], 23.9[18.4], 24.6[18.1], 24.8[30.3], 25.3[94.5], 25.8[ 14.6.5 ], 23.5[ 30.7 ], 23.9[18.4], 24.1, 24.8[30.3], 25.5 ], 25.8[ 94.5, 8[ 14.6.5, 26.6.5 [ 28, 8.6.9, 7, 6.9, 30.6, 30.5 [ 30.8[ 30.5 ], 6.8 [ 30.8, 6.8 [ 30.5, 6, 6.6.8 [ 30..

Single crystal X-ray data are shown in table 3.

TABLE 3-E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino]Single crystal X-ray data for-quinazolin-6-yl } -allyl } -acetamide dimaleate

	Dimaleate salt
		Empirical molecular weight crystal size (mm) space group unit cell size Z (per formula) density (g/cm)3)R	C27H29N5O3 2+·2(C4H3O4 -) 701.680.03X 0.04X 0.20P-1 IIIAs a-4.7763 (4) * b-19.0308 (14) * c-19.1520 (14) * α -100.4 ° β -90.2 ° γ -95.3 ° 21.3670.0648

Example 6

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

the monophosphate salt is prepared as follows. 5.022g E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, prepared according to the method in example 1, 2 or 3 above, was dissolved in 30ml ethanol and heated to 35 ℃ to give a clear solution to prepare a free base solution. 1 equi-molar phosphoric acid (87%, 0.77ml) was diluted with 20ml ethanol. The acid solution was added dropwise to the free base ethanol solution with stirring and heated (-45 to 55 ℃). A yellow precipitate appeared immediately. The slurry became viscous over time, 50ml of ethyl acetate was added and the slurry was allowed to cool to room temperature. The yellow crystalline powder was collected by filtration and dried in vacuo for 2 hours. The yield of the monophosphate product was about 84%. The monophosphate may comprise 1-3% water.

The powder X-ray diffraction pattern of the monophosphate (monohydrate) is shown in FIG. 3. Characteristic X-ray diffraction peaks (monohydrate) of monophosphate (2 θ (± 0.1 °), [% relative intensity ]): 4.9[100], 6.5[2.7], 10.8[2.6], 13.1[3], 14.3[2], 14.9[4.8], 15.5[25.1], 16.3[2.5], 16.7[2.9], 17.2[4.5], 17.9[2.1], 19.9[17.3], 20.6[8.2], 21.7[4.5], 22.1[2], 22.8[2.4], 23.7[3.1], 24.3[1.9], 25.0[8.7], 26.0[3], 26.5[3.9], 27.5[2.2], 28.3[1.8], 29.1[2.1], 30.1[2.2], 35.5[1.6] and 37.7[1.6 ].

Example 7

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

a THF free base solution was prepared by dissolving 104mg of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide, prepared according to the method of example 1, 2 or 3, in 5ml of THF with stirring to give a clear solution. Citraconic acid solution was prepared by dissolving 64mg citraconic acid (about 2.2 equivalents) in 1ml THF. The citraconic acid solution was added dropwise to the free base solution with stirring. After the addition, no precipitate was produced. The volume of the solution was reduced in a nitrogen sparge and then stirred while capped. After about 15 minutes, traces of precipitate were produced. After 1 hour, the solution became a viscous slurry and the slurry was stirred overnight. The precipitate was then separated by vacuum filtration through a 0.45 μm nylon-66 membrane filter.

The prepared solid was rinsed with a few ml of THF and dried under nitrogen. The yield was about 62%.

The product was E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide dicitraconate according to combustion analysis.

Example 8

Synthesis of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide mono malate:

E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide (1 g), prepared according to the method of example 1, 2 or 3, was dissolved in 25ml of hot THF. Malic acid (571 mg; 2 molar equivalents of free base) was added to the free base solution. The mixture was stirred overnight during which time a solid precipitated. An additional 25ml of THF was added, the slurry was stirred for an additional day, and the solid was collected by vacuum filtration to yield the product mono malate complex.

Powder X-ray diffraction showed the material to be crystalline.

Example 9

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

E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide (2 g), prepared according to the method of example 1, 2 or 3, was dissolved in a refluxing 16: 1(v/v) mixture of ethyl acetate (160 ml)/dichloromethane (10 ml). A fumaric acid solution was prepared by dissolving 2 equivalents (1 gram) of fumaric acid in hot ethanol (12 ml). The acid solution was added hot to the refluxing free base solution.

The resulting mixture was stirred and refluxed for about ten minutes, then cooled to room temperature. Hexane (. about.100 ml) was added until the reaction mixture became turbid. The mixture was then sonicated until crystals were produced. The reaction mixture was heated to about 70 ℃ and stirred overnight to give a slurry. The solid was then collected by cold filtration to give the product.

The fumarate salt was a monofumarate hemipentahydrate (2.5H) as determined by elemental analysis₂O)。

Example 10

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

the E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide edisylate complex was prepared by dissolving 0.5 equivalents of 1, 2-ethanedisulfonic acid in 80: 20 Methyl Ethyl Ketone (MEK)/methanol (MeOH) (v/v). E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide free base, prepared according to the method of example 1, 2 or 3, was dissolved in about 60: 40MEK/MeOH (v/v), and 1, 2-ethanedisulfonic acid solution was added dropwise with stirring. Initially an oil is formed which later crystallizes into a solid powder.

Elemental analysis determined the material to be anhydrous hemiedisylate.

Example 11

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

several racemic E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide tartrates were prepared. The synthesis of mono-and hemitartrate hemihydrate was started with the preparation of amorphous material. This material was synthesized by dissolving 3g E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide free base in 20: 3(v/v) ethanol (EtOH)/dichloromethane (50 ml). A solution of D, L-tartaric acid was prepared by dissolving 2 grams of D, L-tartaric acid in 10ml of water. The two solutions were combined and stirred at room temperature for-30 minutes. Solvent reduction, amorphous material is produced.

Example 12

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

simultaneously synthesize the racemate of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridine-3-yloxy) -phenylamino ] -quinazoline-6-yl } -allyl) -acetamide and (+) -10 camphorsulfonic acid complex.

The (+) -10-camphorsulfonic acid complex was synthesized by dissolving 2 grams of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide free base, prepared according to the method of example 1, 2 or 3, in 5: 2(v/v) EtOH/dichloromethane. (+) -10 Camphorsulfonic acid solution was prepared by dissolving 1 gram of (+) -10 Camphorsulfonic acid in 5mL EtOH. The acid solution was added to the free base solution with stirring at room temperature. The reaction mixture was stirred at room temperature for twenty minutes and then the volume of the solution was reduced to yield a crude solid. Part of the crude solid produced was dissolved in hot EtOAc. Hexane was added until turbid, and the mixture was then cooled to room temperature. The solution was sonicated until a precipitate was formed, and then the slurry was left overnight at room temperature. The material was isolated by filtration to yield a yellow solid.

The remaining crude solid above was dissolved in hot EtOAc (75 mL). The solution was cooled and seeded with the yellow solid described above. The reaction mixture was then heated to-75 ℃ and the slurry was left overnight. The mixture was cooled to RT, filtered and washed with EtOAc to give (+) -E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide camsylate.

The racemic camsylate complex was prepared from 1 gram of E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide free base prepared according to the method of example 1 or 2 dissolved in refluxing EtOAc. The acid solution was prepared from 1 gram (. + -.) -10 camphorsulfonic acid in 15ml EtOAc. The acid solution is added to the refluxing free base solution. The solution was refluxed overnight and then isolated by filtration. The solid was then washed with EtOAc and dried to give racemic E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide camsylate. Both racemic and (+) -d-camphorsulfonic acid samples absorbed moisture to the point of deliquescence.

Example 13

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

E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide monobenzenesulfonate was prepared as follows. E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide free base, prepared according to the method of any of examples 1, 2 or 3, 500mg was dissolved in THF. Benzenesulfonic acid (168mg, 1 molar equivalent) was added to the free base solution. Diethyl ether was then added dropwise to the solution until turbidity was observed. After stirring overnight, the precipitate separated as an oil on the flask side. The oily material was scraped off and stirred for another day. The crystalline material was collected after two days.

With DSC, the monobenzenesulfonate started melting at 135 ℃ and the melting point peak was 137 ℃. The hygroscopicity of the substances was evaluated in a relative humidity chamber. After 16 hours in the 75% RH room, the water uptake was insignificant. In the 94% RH chamber, the weight gain was 6.7% after the same time, and deliquescence was observed after 16 hours in the 100% relative humidity chamber.

Example 14

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

E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide free base, prepared according to the method of any of examples 1, 2 or 3 (3.00 g), was dissolved in 40ml ethanol and 6ml dichloromethane. 2.05 molar equivalents of ethanesulfonic acid dissolved in 10mL of ethanol were added to the free base solution. The solution was concentrated, treated in a minimum volume of ethanol, and then ethyl acetate was added as a non-solvent until precipitation occurred. The slurry was stirred at room temperature for more than 48 hours and isolated to give E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide disulfonate.

The diethylsulforate complex was crystalline as determined by PXRD. DSC showed complete melting starting at 146 ℃ and a peak at 149.5 ℃. Moisture absorption: in 90% relative humidity, 45% by weight.

Example 15

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

E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide free base (100mg), prepared according to the method of any of examples 1, 2 or 3, was dissolved in THF and 2 molar equivalents of nitric acid were added. A pale yellow solid precipitated and was isolated to give the product.

Sample E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide dinitrate was crystalline as determined by PXRD. The DSC thermogram shows a sharp exothermic curve at an initial temperature of 148 ℃ and a temperature peak of 151 ℃. Moisture absorption: at 90% relative humidity, 7% by weight.

While the invention has been illustrated in its preferred embodiments, it will be understood that: various modifications thereof will be apparent to those skilled in the art upon reading the specification. It is, therefore, to be understood that the disclosure herein is intended to cover such modifications as fall within the scope of the appended claims.

Claims (14)

1. A compound selected from E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide hydrochloride, E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide maleate, or E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -acetamide Complexes of yl-quinazolin-6-yl } -allyl) -acetamide phosphate.

2. The complex of claim 1, wherein the complex is crystalline.

3. The complex of claim 1, wherein the complex is amorphous.

4. The complex of claim 1, wherein the complex is a dimaleate.

5. The complex of claim 4 wherein the dimaleate salt exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees (2 θ) at about the following:

2θ

6.0

25.2

27.1

27.7

31.4

6. the complex of claim 1, wherein the complex is a monohydrochloride.

7. The complex of claim 6 wherein the monohydrochloride exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees (2 θ) at about:

2θ

4.6

9.3

17.1

18.4

27.5

8. the complex of claim 1, wherein the complex is a monophosphate.

9. The complex of claim 8 wherein the monophosphate exhibits an X-ray powder diffraction pattern having characteristic peaks expressed in degrees (2 Θ) at about the following:

2θ

4.9

15.5

19.9

20.6

25.0

10. a method of inhibiting abnormal cell growth in a mammal comprising administering to said mammal an amount of a compound of claim 1 effective to inhibit abnormal cell growth.

11. The method according to claim 10 wherein the abnormal cell growth is cancer.

12. A method of inhibiting abnormal cell growth in a mammal comprising administering to said mammal an effective abnormal cell growth inhibiting amount of a compound of claim 1 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-hormonal agents, and anti-androgens.

13. A pharmaceutical composition comprising an amount of a compound of claim 1 effective to treat hyperproliferative disorders in a mammal, and a pharmaceutically acceptable carrier.

14. A complex formed by contacting E-2-methoxy-N- (3- {4- [ 3-methyl-4- (6-methyl-pyridin-3-yloxy) -phenylamino ] -quinazolin-6-yl } -allyl) -acetamide with an acid or a reactive equivalent of said acid, wherein said acid is selected from at least one of maleic acid, hydrochloric acid and phosphoric acid.