WO2019071351A1

WO2019071351A1 - Novel fluorinated 4-aryloxyquinazoline derivatives as egfr inhibitors useful for treating cancers

Info

Publication number: WO2019071351A1
Application number: PCT/CA2018/051282
Authority: WO
Inventors: Abdelmalik Slassi; Peter Dove; David Alexander ROSA
Original assignee: Trillium Therapeutics ULC
Current assignee: Trillium Therapeutics ULC
Priority date: 2017-10-12
Filing date: 2018-10-11
Publication date: 2019-04-18
Anticipated expiration: 2020-04-12

Abstract

A novel class of fluorinated derivatives of Formula I have been prepared and found to be useful in the treatment of cancers and other EGFR related disorders. The compounds of Formula I display excellent brain penetration and an improved side effect profile in comparison to previously known compounds (e.g.) erlotinib.

Description

TITLE: NOVEL FLUORINATED 4-ARYLOXYQUINAZOLINE DERIVATIVES AS EGFR INHIBITORS USEFUL FOR TREATING CANCERS

RELATED APPLICATION

[0001 ] The present application claims the benefit of priority from U.S. provisional patent application S.N. 62/571 ,441 filed on October 12, 2017, the contents of which are incorporated herein by reference in their entirety.

FIELD

[0001] The present application relates to novel fluorinated derivatives, to processes for their preparation, to compositions comprising them, and to their use in therapy. More particularly, it relates to compounds useful in the treatment of diseases, disorders or conditions mediated by epidermal growth factor receptor. Such compounds and salts thereof may be useful in the treatment or prevention of a number of different cancers. The application also relates to pharmaceutical compositions comprising said compounds and salts thereof, especially useful polymorphic forms of these compounds and salts, intermediates useful in the manufacture of said compounds and to methods of treatment of diseases mediated by various different forms of EGFR using said compounds and salts thereof.

BACKGROUND

[0002] Epidermal Growth Factor Receptor (EGFR) is a transmembrane protein tyrosine kinase of the ErbB receptor family. Upon binding of a growth factor ligand such as epidermal growth factor (EGF), the receptor can homo- dimerise with another EGFR molecule or hetero-dimerise with another family member such as ErbB2 (HER2), ErbB3 (HER3), or ErbB4 (HER4). Homo- and/or hetero-dimerisation of ErbB receptors results in the phosphorylation of key tyrosine residues in the intracellular domain and leads to the stimulation of numerous intracellular signal transduction pathways involved in cell proliferation and survival. Deregulation of ErbB family signalling promotes proliferation, invasion, metastasis, angiogenesis, and tumour cell survival and has been described in many human cancers, including those of the lung, head, neck and breast. The ErbB family therefore represents a rational target for anticancer drug development and a number of agents targeting EGFR or ErbB2 are now clinically available, including gefitinib (IRESSA™), erlotinib (TARCEVA™) and lapatinib (TYKERB™, TYVERB™). Detailed reviews of ErbB receptor signalling and its involvement in tumourigenesis are provided in New England Journal of Medicine (2008) Vol. 358, 1 160-74 and Biochemical and Biophysical Research Communications (2004) Vol. 319, l-ll. In 2004 it was reported (Science [2004] Vol .304, 1497-500 and New England Journal of Medicine [2004] Vol. 350,2129-39) that activating mutations in EGFR correlated with response to gefitinib therapy in non-small-cell lung cancer (NSCLC).

[0003] The most common EGFR activating mutations, L858R and de1 E746_A750, result in an increase in affinity for small molecule tyrosine kinase inhibitors such as gefitinib and erlotinib and a decrease in affinity for adenosine triphosphate (ATP) relative to wild type (WT) EGFR. Ultimately, acquired resistance to therapy with gefitinib or erlotinib arises, for example by mutation of the gatekeeper residue T790M, which is reportedly detected in 50% of clinically resistant patients. This mutation is not believed to hinder the binding of gefitinib or erlotinib to EGFR sterically, it merely alters the affinity to ATP to levels comparable to WT EGFR. In view of the importance of this mutation in resistance to existing therapies targeting EGFR, agents which inhibit EGFR harbouring the gatekeeper mutation may be especially useful in the treatment of cancer. There remains a need for compounds that exhibit favourable potency against WT EGFR versus activating mutant forms of EGFR (for example the L858R EGFR mutant, or the del E746-A750 mutant or the Exon19 deletion EGFR mutant) and/or resistant mutant forms of EGFR (for example T790M EGFR mutant), and/or selectivity over other enzyme receptors. For example, the human ether-a-go-go related gene (hERG) channels encode the pore- forming subunit of the inward rectifying voltage gated potassium channel expressed in the heart (kr) as well as in brain regions and endocrine cells. This subunit is involved in cardiac action potential repolarization, and reduced function of hERG lengthens ventricular action potentials amd prolongs the QT interval in an electrocardiogram. Blockade of this target increases the risk for potentially fatal ventricular arrhythmia therefore it is desirable for compounds that are to act as drugs to have minimal potency at the hERG channel [Physiological Reviews 2012 92(3) 1393} In addition, there remains a need for compounds that can effectively penetrate the blood brain barrier. Such compounds may be expected to be more suitable as therapeutic agents, particularly for the treatment of cancers of the brain.

[0004] Glioblastoma multiforme (GBM) is the most aggressive of the astrocytic malignancies and the most common intracranial tumor in adults. Although the EGFR is overexpressed and/or mutated in at least 50% of GBM cases and is required for tumor maintenance in animal models, EGFR inhibitors have thus far failed to deliver significant responses in GBM patients. One inherent resistance mechanism in GBM is the coactivation of multiple receptor tyrosine kinases, which generates redundancy in activation of phosphoinositide-3'-kinase (PI3K) signaling. Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) tumor suppressor is frequently phosphorylated at a conserved tyrosine residue, Y240, in GBM clinical samples. Phosphorylation of Y240 is associated with shortened overall survival and resistance to EGFR inhibitor therapy in GBM patients and plays an active role in mediating resistance to EGFR inhibition in vitro. Y240 phosphorylation can be mediated by both fibroblast growth factor receptors and SRC family kinases (SFKs) but does not affect the ability of PTEN to antagonize PI3K signaling. These findings show that, in addition to genetic loss and mutation of PTEN, its modulation by tyrosine phosphorylation has important implications for the development and treatment of GBM.

SUMMARY

[0005] A novel class of fluorinated derivatives of Formula I has been prepared and found to be useful in the treatment of cancers and other EGFR related disorders. Surprisingly, it was observed that replacement of a nitrogen atom in a previously known class of non-covalent EGFR inhibitors with an oxygen atom retained biochemical activity and showed reduced hERG channel potency.

[0006] The compound(s) of the application also exhibit favourable toxicity profiles (for example a decreased hERG blocking liability) in comparison with other known EGFR / EGFR-mutant inhibitors. Therefore, in some embodiments, the compounds of the application are especially useful in the treatment of disease states in which EGFR and/or activating mutations of EGFR and/or resistance mutations of EGFR are implicated, for example in the treatment of cancer.

[0007] Accordingly, the present application includes a compound of Formula I or a pharmaceutically acceptable salt and/or solvate thereof:

I wherein:

R¹ is selected from unsubstituted or substituted phenyl and unsubstituted or substituted naphthyl, wherein the substituents for R¹ are selected from one or more of CI, Br, F, Ci-₄alkyl, C2-4alkenyl, C2-4alkynyl, Ci-₄fluoroalkyl, C2- ₄fluoroalkenyl and C2-4fluoroalkynyl;

R² is selected from (CH₂)nOR⁴ and C(0)X(CH₂)nOR⁴;

R³ is selected from Ci-₄alkyl, Ci-₄fluoroalkyl and (CH2)mOR⁵;

R⁴ is selected from CHF2 and CF3;

R⁵ is selected from CHF2, CF3 and CH3;

X is selected from NH, NCH3 and O;

n is selected from 1 , 2, 3 and 4; and

m is selected from 1 , 2, 3, and 4.

[0008] The present application also includes a composition comprising one or more compounds of the application and a carrier. In an embodiment, the composition is a pharmaceutical composition comprising one or more compounds of the application and a pharmaceutically acceptable carrier.

[0009] The compounds of the application have been shown to be capable of inhibiting EGFR protein function. Therefore the compounds of the application are useful for treating diseases, disorders or conditions treatable by inhibition of EGFR. Accordingly, the present application also includes a method of treating a disease, disorder or condition treatable by inhibition of EGFR, comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof.

[0010] In a further embodiment, the compounds of the application are used as medicaments. Accordingly, the application also includes a compound of the application for use as a medicament.

[0011] The present application also includes a use of one or more compounds of the application for treatment of a disease, disorder or condition treatable by inhibition of EGFR as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of a disease, disorder or condition treatable by inhibition of EGFR. The application further includes one or more compounds of the application for use in treating a disease, disorder or condition treatable by inhibition of EGFR.

[0012] In an embodiment, the disease, disorder or condition treatable by inhibition of EGFR is a neoplastic disorder. In an embodiment, the treatment is in an amount effective to ameliorate at least one symptom of the neoplastic disorder, for example, reduced cell proliferation or reduced tumor mass in a subject in need of such treatment.

[0013] In an embodiment, the disease, disorder or condition is cancer.

[0014] In an embodiment, the disease, disorder or condition is a disease, disorder or condition associated with an uncontrolled and/or abnormal cellular activity affected directly or indirectly by EGFR. In another embodiment, the uncontrolled and/or abnormal cellular activity that is affected directly or indirectly by EGFR is proliferative activity in a cell.

[0015] The application also includes a method of inhibiting proliferative activity in a cell, comprising administering an effective amount of one or more compounds of the application to the cell.

[0016] In a further embodiment the disease, disorder or condition treatable by inhibition of EGFR is cancer and the one or more compounds of the application are administered in combination with one or more additional cancer treatments. In another embodiment, the additional cancer treatment is selected from radiotherapy, chemotherapy, targeted therapies such as antibody therapies and small molecule therapies such as tyrosine-kinase inhibitors, immunotherapy, hormonal therapy and anti-angiogenic therapies.

[0017] The application additionally provides a process for the preparation of compounds of the applicaition. General and specific processes are discussed in more detail below and set forth in the Examples.

[0018] Other features and advantages of the present application will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating embodiments of the application, are given by way of illustration only and the scope of the claims should not be limited by these embodiments, but should be given the broadest interpretation consistent with the description as a whole.

DETAILED DESCRIPTION

Definitions

[0019] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the application herein described for which they are suitable as would be understood by a person skilled in the art. Unless otherwise specified within this application or unless a person skilled in the art would understand otherwise, the nomenclature used in this application generally follows the examples and rules stated, for example, in "Nomenclature of Organic Chemistry" (Pergamon Press, 1979), Sections A, B, C, D, E, F, and H. Optionally, a name of a compound may be generated using a chemical naming program such as ACD/ChemSketch, Version 5.09/September 2001 , Advanced Chemistry Development, Inc., Toronto, Canada.

[0020] The term "compound of the application" or "compound of the present application" and the like as used herein refers to a compound of Formula I, and pharmaceutically acceptable salts, solvates, prodrugs and/or radiolabeled versions thereof. [0021] The term "composition of the application" or "composition of the present application" and the like as used herein refers to a composition, such as a pharmaceutical composition, comprising one or more compounds of Formula I , or pharmaceutically acceptable salts, solvates, prodrugs and/or radiolabeled versions thereof .

[0022] The term "and/or" as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that "at least one of" or "one or more" of the listed items is used or present. The term "and/or" with respect to pharmaceutically acceptable salts, solvates and/or prodrugs thereof means that the compounds of the application exist as individual salts, hydrates or prodrugs, as well as a combination of, for example, a salt of a solvate of a compound of the application or a salt of a prodrug of a compound of a compound of the application.

[0023] As used in the present application, the singular forms "a", "an" and "the" include plural references unless the content clearly dictates otherwise. For example, an embodiment including "a compound" should be understood to present certain aspects with one compound, or two or more additional compounds.

[0024] In embodiments comprising an "additional" or "second" component, such as an additional or second compound, the second component as used herein is chemically different from the other components or first component. A "third" component is different from the other, first, and second components, and further enumerated or "additional" components are similarly different.

[0025] As used in the present application, the singular forms "a", "an" and "the" include plural references unless the content clearly dictates otherwise. For example, an embodiment including "a compound" should be understood to present certain aspects with one compound, or two or more additional compounds.

[0026] In embodiments comprising an "additional" or "second" component, such as an additional or second compound, the second component as used herein is chemically different from the other components or first component. A "third" component is different from the other, first, and second components, and further enumerated or "additional" components are similarly different.

[0027] In understanding the scope of the present application, the term "comprising" (and any form of comprising, such as "comprise" and "comprises"), "having" (and any form of having, such as "have" and "has"), "including" (and any form of including, such as "include" and "includes") or "containing" (and any form of containing, such as "contain" and "contains"), are inclusive or openended terms and do not exclude additional, unrecited elements or process steps.

[0028] The term "consisting" and its derivatives as used herein are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and/or steps, and also exclude the presence of other unstated features, elements, components, groups, integers and/or steps.

[0029] The term "consisting essentially of" as used herein is intended to specify the presence of the stated features, elements, components, groups, integers, and/or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and/or steps.

[0030] The term "suitable" as used herein means that the selection of the particular compound or conditions would depend on the specific synthetic manipulation to be performed, the identity of the molecule(s) to be transformed and/or the specific use for the compound, but the selection would be well within the skill of a person trained in the art.

[0031] In embodiments of the present application, the compounds described herein may have at least one asymmetric center. Where compounds possess more than one asymmetric center, they may exist as diastereomers. It is to be understood that all such isomers and mixtures thereof in any proportion are encompassed within the scope of the present application. It is to be further understood that while the stereochemistry of the compounds may be as shown in any given compound listed herein, such compounds may also contain certain amounts (for example, less than 20%, suitably less than 10%, more suitably less than 5%) of compounds of the present application having alternate stereochemistry. It is intended that any optical isomers, as separated, pure or partially purified optical isomers or racemic mixtures thereof are included within the scope of the present application.

[0032] The compounds of the present application may also exist in different tautomeric forms and it is intended that any tautomeric forms which the compounds form, as well as mixtures thereof, are included within the scope of the present application.

[0033] The compounds of the present application may further exist in varying polymorphic forms and it is contemplated that any polymorphs, or mixtures thereof, which form are included within the scope of the present application.

[0034] Terms of degree such as "substantially", "about" and "approximately" as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies or unless the context suggests otherwise to a person skilled in the art.

[0035] The term "alkyl" as used herein, whether it is used alone or as part of another group, means straight or branched chain, saturated alkyl groups. The number of carbon atoms that are possible in the referenced alkyl group are indicated by the prefix "Cni-n2". For example, the term Ci-6alkyl means an alkyl group having 1 , 2, 3, 4, 5 or 6 carbon atoms.

[0036] The term "alkylene", whether it is used alone or as apart of another group, means straight or branched chain, saturated alkylene group, that is, a saturated carbon chain that contains substituents on two of its ends. The number of carbon atoms that are possible in the referenced alkylene group are indicated by the prefix "Cn-iV. For example, the term Ci-6alkylene means an alkylene group having 1 , 2, 3, 4, 5 or 6 carbon atoms.

[0037] The term "alkenyl" as used herein, whether it is used alone or as part of another group, means straight or branched chain, unsaturated alkyl groups containing at least one double bond. The number of carbon atoms that are possible in the referenced alkylene groups are indicated by the prefix "Cn-i- n2". For example, the term C2-6alkenyl means an alkenyl group having 2, 3, 4, 5 or 6 carbon atoms and at least one double bond.

[0038] The term "alkynyl" as used herein, whether it is used alone or as part of another group, means straight or branched chain unsaturated alkyl groups containing at least one triple bond. The number of carbon atoms that are possible in the referenced alkylyne group are indicated by the prefix "Cn-i- n2". For example, the term C2-6alkynyl means an alkynyl group having 2, 3, 4, 5 or 6 carbon atoms and at least one triple bond.

[0039] The term "fluoroalkyl" as used herein refers to an alkyl group wherein one or more, including all of the hydrogen atoms are replaced by a fluorine atom.

[0040] As a prefix, the term "substituted" as used herein refers to a structure, molecule or group in which one or more available hydrogen atoms are replaced with one or more other chemical groups.

[0041] The term "available", as in "available hydrogen atoms" or "available atoms" refers to atoms that would be known to a person skilled in the art to be capable of replacement by a substituent.

[0042] The term "optionally substituted" refers to groups, structures, or molecules that are either unsubstituted or are substituted with one or more substituents.

[0043] The term "protecting group" or "PG" and the like as used herein refers to a chemical moiety which protects or masks a reactive portion of a molecule to prevent side reactions in those reactive portions of the molecule, while manipulating or reacting a different portion of the molecule. After the manipulation or reaction is complete, the protecting group is removed under conditions that do not degrade or decompose the remaining portions of the molecule. The selection of a suitable protecting group can be made by a person skilled in the art. Many conventional protecting groups are known in the art, for example as described in "Protective Groups in Organic Chemistry" McOmie, J.F.W. Ed., Plenum Press, 1973, in Greene, T.W. and Wuts, P.G.M., "Protective Groups in Organic Synthesis", John Wiley & Sons, 3^rd Edition, 1999 and in Kocienski, P. Protecting Groups, 3rd Edition, 2003, Georg Thieme Verlag (The Americas).

[0044] The term "cell" as used herein refers to a single cell or a plurality of cells and includes a cell either in a cell culture or in a subject.

[0045] The term "subject" as used herein includes all members of the animal kingdom including mammals, and suitably refers to humans. Thus the methods of the present application are applicable to both human therapy and veterinary applications. In an embodiment, the subject is a mammal. In another embodiment, the subject is human.

[0046] The term "pharmaceutically acceptable" means compatible with the treatment of subjects, for example humans.

[0047] The term "pharmaceutically acceptable carrier" means a nontoxic solvent, dispersant, excipient, adjuvant or other material which is mixed with the active ingredient in order to permit the formation of a pharmaceutical composition, i.e., a dosage form capable of administration to a subject. One non-limiting example of such a carrier is a pharmaceutically acceptable oil typically used for parenteral administration.

[0048] The term "pharmaceutically acceptable salt" means either an acid addition salt or a base addition salt which is suitable for, or compatible with the treatment of subjects.

[0049] An acid addition salt suitable for, or compatible with, the treatment of subjects is any non-toxic organic or inorganic acid addition salt of any basic compound. Basic compounds that form an acid addition salt include, for example, compounds comprising an amine group. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric, nitric and phosphoric acids, as well as acidic metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids which form suitable salts include mono-, di- and tricarboxylic acids. Illustrative of such organic acids are, for example, acetic, trifluoroacetic, propionic, glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, benzoic, hydroxybenzoic, phenylacetic, cinnamic, mandelic, salicylic, 2-phenoxybenzoic, p-toluenesulfonic acid and other sulfonic acids such as methanesulfonic acid, ethanesulfonic acid and 2- hydroxyethanesulfonic acid. Either the mono- or di-acid salts can be formed, and such salts can exist in either a hydrated, solvated or substantially anhydrous form. In general, acid addition salts are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection criteria for the appropriate salt will be known to one skilled in the art. Other non-pharmaceutically acceptable salts such as but not limited to oxalates may be used, for example in the isolation of compounds of the application for laboratory use, or for subsequent conversion to a pharmaceutically acceptable acid addition salt.

[0050] A base addition salt suitable for, or compatible with, the treatment of subjects is any non-toxic organic or inorganic base addition salt of any acidic compound. Acidic compounds that form a basic addition salt include, for example, compounds comprising a carboxylic acid group. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium or barium hydroxide as well as ammonia. Illustrative organic bases which form suitable salts include aliphatic, alicyclic or aromatic organic amines such as isopropylamine, methylamine, trimethylamine, picoline, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2- diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N- ethylpiperidine, polyamine resins, and the like. Exemplary organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. [See, for example, S. M. Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1 -19]. The selection of the appropriate salt may be useful so that an ester functionality, if any, elsewhere in a compound is not hydrolyzed. The selection criteria for the appropriate salt will be known to one skilled in the art.

[0051] The term "solvate" as used herein means a compound, or a salt or prodrug of a compound, wherein molecules of a suitable solvent are incorporated in the crystal lattice. A suitable solvent is physiologically tolerable at the dosage administered. Examples of suitable solvents are ethanol, water and the like. When water is the solvent, the molecule is referred to as a "hydrate". The formation of solvates of the compounds of the application will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions. The selection of suitable conditions to form a particular solvate can be made by a person skilled in the art.

[0052] The term "treating" or "treatment" as used herein and as is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. not worsening) state of disease, preventing spread of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission (whether partial or total), whether detectable or undetectable. "Treating" and "treatment" can also mean prolonging survival as compared to expected survival if not receiving treatment. "Treating" and "treatment" as used herein also include prophylactic treatment. For example, a subject with early cancer can be treated to prevent progression, or alternatively a subject in remission can be treated with a compound or composition described herein to prevent recurrence. Treatment methods comprise administering to a subject a therapeutically effective amount of one or more of the compounds of the application and optionally consist of a single administration, or alternatively comprise a series of administrations. For example, the compounds of the application are administered at least once a week. However, in another embodiment, the compounds are administered to the subject from about one time per two weeks, three weeks or one month. In another embodiment, the compounds are administered about one time per week to about once daily. In another embodiment, the compounds are administered 2, 3, 4, 5 or 6 times daily. The length of the treatment period depends on a variety of factors, such as the severity of the disease, disorder or condition, the age of the subject, the concentration and/or the activity of the compounds of the application, and/or a combination thereof. It will also be appreciated that the effective dosage of the compound used for the treatment may increase or decrease over the course of a particular treatment regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration may be required. For example, the compounds are administered to the subject in an amount and for duration sufficient to treat the patient.

[0053] "Palliating" a disease, disorder or condition means that the extent and/or undesirable clinical manifestations of a disease, disorder or condition are lessened and/or time course of the progression is slowed or lengthened, as compared to not treating the disorder.

[0054] The term "prevention" or "prophylaxis", or synonym thereto, as used herein refers to a reduction in the risk or probability of a patient becoming afflicted with a disease, disorder or condition or manifesting a symptom associated with a disease, disorder or condition.

[0055] The "disease, disorder or condition" as used herein refers to a disease, disorder or condition treatable by inhibition of EGFR activity and particularly using an EGFR inhibitor, such as a compound of the application herein described.

[0056] The term "treatable by inhibition of EGFR" as used herein means that the disease, disorder or condition to be treated is affected by, modulated by and/or has some biological basis, either direct or indirect, that includes aberrant EGFR activity, in particular, increased EGFR activity or, also, decreased EGFR activity such as results from mutation or splice variation and the like. These diseases respond favourably when EGFR activity associated with the disease is blocked or inhibited by one or more of the present compounds.

[0057] As used herein, the term "effective amount" or "therapeutically effective amount" means an amount of one or more compounds of the application that is effective, at dosages and for periods of time necessary to achieve the desired result. For example in the context of treating a disease, disorder or condition, an effective amount is an amount that, for example, inhibits EGFR activity compared to the inhibition without administration of the one or more compounds. In an embodiment, effective amounts vary according to factors such as the disease state, age, sex and/or weight of the subject. In a further embodiment, the amount of a given compound or compounds that will correspond to an effective amount will vary depending upon factors, such as the given drug(s) or compound(s), the pharmaceutical formulation, the route of administration, the type of condition, disease or disorder, the identity of the subject being treated, and the like, but can nevertheless be routinely determined by one skilled in the art. The effective amount is one that following treatment therewith manifests as an improvement in or reduction of any disease symptom. When the disease is cancer, amounts that are effective can cause a reduction in the number, growth rate, size and/or distribution of tumours.

[0058] The term "administered" as used herein means administration of a therapeutically effective amount of one or more compounds or compositions of the application to a cell either in cell culture or in a subject.

[0059] The term "neoplastic disorder" as used herein refers to a disease, disorder or condition characterized by cells that have the capacity for autonomous growth or replication, e.g., an abnormal state or condition characterized by proliferative cell growth. The term "neoplasm" as used herein refers to a mass of tissue resulting from the abnormal growth and/or division of cells in a subject having a neoplastic disorder. Neoplasms can be benign (such as uterine fibroids and melanocytic nevi), potentially malignant (such as carcinoma in situ) or malignant (i.e. cancer). Exemplary neoplastic disorders include but are not limited to carcinoma, sarcoma, metastatic disorders (e.g., tumors arising from the prostate), hematopoietic neoplastic disorders, (e.g., leukemias, lymphomas, myeloma and other malignant plasma cell disorders), metastatic tumors and other cancers.

[0060] The term "cancer" as used herein refers to cellular-proliferative disease states.

II. Compounds and Compositions of the Application

[0061] Compounds of the present application were prepared and were found to inhibit uncontrolled and/or abnormal cellular activities affected directly or indirectly by EGFR protein. In particular, compounds of the present application exhibited activity as EGFR inhibitors, and are therefore useful in therapy, for example for the treatment of neoplastic disorders such as cancer. Compounds of the application also showed reduced inhibition of hERG compared to closely related arylaminequinazoline compounds indicating that compounds of the application may show a reduced risk for cardiovascular side effects.

[0062] Accordingly, the present application includes a compound of Formula I or a pharmaceutically acceptable sal or solvate:

wherein:

R² is selected from (CH₂)nOR⁴ and C(0)X(CH₂)nOR⁴;

R³ is selected from Ci-₄alkyl, Ci-₄fluoroalkyl and (CH2)mOR⁵;

R⁴ is selected from CHF2 and CF3;

R⁵ is selected from CHF2, CF3 and CH3;

X is selected from NH, NCH3 and O;

n is selected from 1 , 2, 3 and 4; and

m is selected from 1 , 2, 3, and 4.

[0063] In some embodiments, R¹ has one to three substituents. In some embodiments, R¹ is selected from substituted phenyl, wherein the substituents for R¹ are selected from one to three of CI, Br, F, Ci-₄alkyl, C2-4alkenyl, C2- ₄alkynyl, Ci-₄fluoroalkyl, C2-4fluoroalkenyl and C2-4fluoroalkynyl. In some embodiments, R¹ is selected from substituted phenyl, wherein the substituents for R¹ are selected from one to three of CI, Br, F, CH3, CF3, CH=CH₂ and C≡CH. In some embodiments, R¹ is selected from substituted phenyl, wherein the substituents for R¹ are selected from one or two of CI, Br, F, CH3, CF3, CH=CH2 and C≡CH. In some embodiments the substituents on the phenyl of R¹ are in the ortho or meta positions. In some embodiments, R¹ is selected from:

In some embodiments, both of R² and R³ are . in some embodiments, R² is F and R³ is CH3.

[0065] In some embodiments, R⁴ is CHF2.

[0066] In some embodiments, R⁵ is CHF2.

[0067] In some embodiments, X is O.

[0068] In some embodiments, n is selected from 1 , 2 and 3. In some embodiments, n is 2.

[0069] In some embodiments, m is selected from 1 , 2 and 3. In some embodiments, m is 2.

[0070] In some embodiments, the compounds of Formula I are selected from:

6,7-bis[2-(difluoromethoxy)ethoxy]- 4-(3-ethynylphenoxy)quinazoline

1-8

F

[4-(3-chloro-2-fluoro-phenoxy)-7- methoxy-quinazolin-6-yl] N-[2-

1-9 (difluoromethoxy)ethyl]-N-methyl- carbamate

and pharmaceutically acceptable salts and/or solvates thereof.

[0071] As noted above, all stereoisomers are included within the scope of the present application.

Preparation of Compounds

[0072] Compounds of the present application can be prepared by various synthetic processes. The choice of particular structural features and/or substituents may influence the selection of one process over another. The selection of a particular process to prepare a given compound of Formula I is within the purview of the person of skill in the art. Some starting materials for preparing compounds of the present application are available from commercial chemical sources. Other starting materials, for example as described below, are readily prepared from available precursors using straightforward transformations that are well known in the art.

[0073] In some embodiments, the compounds of Formula I are prepared according to the processes illustrated in Schemes l-V. Variables in these schemes are as defined above for Formula I unless otherwise specified.

[0074] In some embodiments, commercially available 6-benzyloxy-4- chloro-7-methoxy-quinazoline (A) is a versatile intermediate that can be used to make certain compounds of Formula I as shown in Scheme I . Therefore, for example, compounds of Formula A are reacted with phenols or naphthols R¹OH in the presence of a suitable base, such as potassium carbonate, in a suitable solvent, such as as dimethylformamide, to provide compounds of Formula B. Deprotection of compounds of Formula B, for example in the presence of an acid such as trifluoroacetic acid (TFA) provides compounds of Formula C which are reacted with various alkylating or acylating reagents R²LG, wherein R² is as defined in Formula I and LG is a suitable leaving group, such as halo, to provide compounds of Formula I, wherein R³ is CH3.

Scheme I

[0075] In some embodiments, intermediates of Formula C are used to prepare compounds of Formula I wherein R² is C(0)X(CH2)nOR⁴ and X is NH or NCH3 by reacting the compounds of Formula C with phosgene in the presence of a suitable amine as shown in Scheme I I.

Scheme II

[0076] In some embodiments, commercially available 6-benzyloxy-4- chloro-7-methoxy-quinazoline (D) is a versatile intermediate that is used to make certain compounds of Formula I as shown in Scheme II I. Therefore, for example, compounds of Formula D are reacted with phenols or naphthols R¹OH in the presence of a suitable base, such as potassium carbonate, in a suitable solvent, such as as dimethylformamide, to provide compounds of Formula E. Compounds of Formula E are then deprotected, using acidic conditions, for example in the presence of TFA, to give compounds of Formula F. Compounds of Formula F are then reacted with various alkylating or acylating reagents R²LG or R³LG, wherein R² amd R3 are as defined in Formula I and LG is a suitable leaving group, such as halo, to provide compounds of Formula I wherein R² and R³ are the same.

Scheme III

[0077] In some embodiments, known intermediate J (PCT appl. 2000055141 ) is another versatile internmediate that is used to make certain compounds of Formula I as shown in Scheme IV. Therefore, for example, compounds of Formula J are selectively alkylated at the phenolic position using a suitable base, such as potassium carbonate, in a suitable solvent, such as as dimethylformamide, to give compounds of Formula K which are then chlorinated to give compounds of Formula L. Compounds of Formula L then undergo a displacement reaction with various phenols or naphthols (R¹-OH) in the presence of a suitable base, such as potassium carbonate, in a suitable solvent, such as as dimethylformamide, giving compounds of Fornula M. This intermediate is then deprotected using acidic conditions (for example, TFA) to give compounds of Formula N. Compounds of Formula N are then be alkylated with R²LG, wherein LG is a suitable leaving group such as halo, to give the desired compounds of Formula I, where R² and R³ may or may not be identical.

N

R² and R³ are identical or different

Scheme IV

[0078] As is shown in Scheme V, in some embodiments, the substituents on R¹ may be converted using known reactions into other substituents on R¹. In one representative example compounds of Formulat I wherein R¹ is substituted at least with one bromine are cross-coupled to trimethylsilylacetylene using bis(triphenylphosphine)palladium(ll) dichloride to give compounds of Formula O which can be deprotected, for example, using potassium carbonate and methanol to give compounds of Formula I, wherein R¹ is substituted with an acetylene (C2alkynyl).

o

Scheme V [0079] In Schemes l-IV the benzyl group (Ph-Chte) is used as a protecting group. A person skilled in the art would appreciate that other protecting groups may be used in place of the benzyl group. In some embodiments, the other protecting groups are also removable under acidic conditions.

[0080] Throughout the processes described herein it is to be understood that, where appropriate, suitable protecting groups will be added to, and subsequently removed from, the various reactants and intermediates in a manner that will be readily understood by one skilled in the art. Conventional procedures for using such protecting groups as well as examples of suitable protecting groups are described, for example, in "Protective Groups in Organic Synthesis", T.W. Green, P.G.M. Wuts, Wiley-lnterscience, New York, (1999). It is also to be understood that a transformation of a group or substituent into another group or substituent by chemical manipulation can be conducted on any intermediate or final product on the synthetic path toward the final product, in which the possible type of transformation is limited only by inherent incompatibility of other functionalities carried by the molecule at that stage to the conditions or reagents employed in the transformation. Such inherent incompatibilities, and ways to circumvent them by carrying out appropriate transformations and synthetic steps in a suitable order, will be readily understood to one skilled in the art. Examples of transformations are given herein, and it is to be understood that the described transformations are not limited only to the generic groups or substituents for which the transformations are exemplified. References and descriptions of other suitable transformations are given in "Comprehensive Organic Transformations - A Guide to Functional Group Preparations" R.C. Larock, VHC Publishers, Inc. (1989). References and descriptions of other suitable reactions are described in textbooks of organic chemistry, for example, "Advanced Organic Chemistry", March, 4th ed. McGraw Hill (1992) or, "Organic Synthesis", Smith, McGraw Hill, (1994). Techniques for purification of intermediates and final products include, for example, straight and reversed phase chromatography on column or rotating plate, recrystallisation, distillation and liquid-liquid or solid-liquid extraction, which will be readily understood by one skilled in the art. Compositions

[0081] The compounds of the present application are suitably formulated in a conventional manner into compositions using one or more carriers. Accordingly, the present application also includes a composition comprising one or more compounds of the application and a carrier. The compounds of the application are suitably formulated into pharmaceutical compositions for administration to subjects in a biologically compatible form suitable for administration in vivo. Accordingly, the present application further includes a pharmaceutical composition comprising one or more compounds of the application and a pharmaceutically acceptable carrier. In embodiments of the application the pharmaceutical compositions are used in the treatment of nay of the diseases, disorders or conditions described herein.

[0082] The compounds of the application are administered to a subject in a variety of forms depending on the selected route of administration, as will be understood by those skilled in the art. For example, a compound of the application is administered by oral, inhalation, parenteral, buccal, sublingual, nasal, rectal, vaginal, patch, pump, topical or transdermal administration and the pharmaceutical compositions formulated accordingly. In some embodiments, administration is by means of a pump for periodic or continuous delivery. Conventional procedures and ingredients for the selection and preparation of suitable compositions are described, for example, in Remington's Pharmaceutical Sciences (2000 - 20th edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999.

[0083] Parenteral administration includes systemic delivery routes other than the gastrointestinal (Gl) tract, and includes, for example intravenous, intraarterial, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary (for example, by use of an aerosol), intrathecal, rectal and topical (including the use of a patch or other transdermal delivery device) modes of administration. Parenteral administration may be by continuous infusion over a selected period of time. [0084] In some embodiments, a compound of the application is orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it is enclosed in hard or soft shell gelatin capsules, or it is compressed into tablets, or it is incorporated directly with the food of the diet. In some embodiments, the compound is incorporated with excipient and used in the form of ingestible tablets, buccal tablets, troches, capsules, caplets, pellets, granules, lozenges, chewing gum, powders, syrups, elixirs, wafers, aqueous solutions and suspensions, and the like. In the case of tablets, carriers that are used include lactose, corn starch, sodium citrate and salts of phosphoric acid. Pharmaceutically acceptable excipients include binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose); fillers (e.g. , lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g., magnesium stearate, talc or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulphate). In embodiments, the tablets are coated by methods well known in the art. In the case of tablets, capsules, caplets, pellets or granules for oral administration, pH sensitive enteric coatings, such as Eudragits™ designed to control the release of active ingredients are optionally used. Oral dosage forms also include modified release, for example immediate release and timed- release, formulations. Examples of modified-release formulations include, for example, sustained-release (SR), extended-release (ER, XR, or XL), time- release or timed-release, controlled-release (CR), or continuous-release (CR or Contin), employed, for example, in the form of a coated tablet, an osmotic delivery device, a coated capsule, a microencapsulated microsphere, an agglomerated particle, e.g., as of molecular sieving type particles, or, a fine hollow permeable fiber bundle, or chopped hollow permeable fibers, agglomerated or held in a fibrous packet. Timed-release compositions are formulated, for example as liposomes or those wherein the active compound is protected with differentially degradable coatings, such as by microencapsulation, multiple coatings, etc. Liposome delivery systems include, for example, small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. In some embodiments, liposomes are formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines. For oral administration in a capsule form, useful carriers or diluents include lactose and dried corn starch.

[0085] In some embodiments, liquid preparations for oral administration take the form of, for example, solutions, syrups or suspensions, or they are suitably presented as a dry product for constitution with water or other suitable vehicle before use. When aqueous suspensions and/or emulsions are administered orally, the compound of the application is suitably suspended or dissolved in an oily phase that is combined with emulsifying and/or suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents are added. Such liquid preparations for oral administration are prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid). Useful diluents include lactose and high molecular weight polyethylene glycols.

[0086] It is also possible to freeze-dry the compounds of the application and use the lyophilizates obtained, for example, for the preparation of products for injection.

[0087] In some embodiments, a compound of the application is administered parenterally. For example, solutions of a compound of the application are prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. In some embodiments, dispersions are prepared in glycerol, liquid polyethylene glycols, DMSO and mixtures thereof with or without alcohol, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. A person skilled in the art would know how to prepare suitable formulations. For parenteral administration, sterile solutions of the compounds of the application are usually prepared, and the pH's of the solutions are suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic. For ocular administration, ointments or droppable liquids are delivered, for example, by ocular delivery systems known to the art such as applicators or eye droppers. In some embodiment, such compositions include mucomimetics such as hyaluronic acid, chondroitin sulfate, hydroxypropyl methylcellulose or polyvinyl alcohol, preservatives such as sorbic acid, EDTA or benzyl chromium chloride, and the usual quantities of diluents or carriers. For pulmonary administration, diluents or carriers will be selected to be appropriate to allow the formation of an aerosol.

[0088] In some embodiments, a compound of the application is formulated for parenteral administration by injection, including using conventional catheterization techniques or infusion. Formulations for injection are, for example, presented in unit dosage form, e.g., in ampoules or in multi- dose containers, with an added preservative. In some embodiments, the compositions take such forms as sterile suspensions, solutions or emulsions in oily or aqueous vehicles, and contain formulating agents such as suspending, stabilizing and/or dispersing agents. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. Alternatively, the compounds of the application are suitably in a sterile powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0089] In some embodiments, compositions for nasal administration are conveniently formulated as aerosols, drops, gels and powders. For intranasal administration or administration by inhalation, the compounds of the application are conveniently delivered in the form of a solution, dry powder formulation or suspension from a pump spray container that is squeezed or pumped by the patient or as an aerosol spray presentation from a pressurized container or a nebulizer. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container, which, for example, take the form of a cartridge or refill for use with an atomising device. Alternatively, the sealed container is a unitary dispensing device such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve which is intended for disposal after use. Where the dosage form comprises an aerosol dispenser, it will contain a propellant which is, for example, a compressed gas such as compressed air or an organic propellant such as fluorochlorohydrocarbon. Suitable propellants include but are not limited to dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, heptafluoroalkanes, carbon dioxide or another suitable gas. In the case of a pressurized aerosol, the dosage unit is suitably determined by providing a valve to deliver a metered amount. In some embodiments, the pressurized container or nebulizer contains a solution or suspension of the active compound. Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insufflator are, for example, formulated containing a powder mix of a compound of the application and a suitable powder base such as lactose or starch. The aerosol dosage forms can also take the form of a pump-atomizer.

[0090] Compositions suitable for buccal or sublingual administration include tablets, lozenges, and pastilles, wherein a compound of the application is formulated with a carrier such as sugar, acacia, tragacanth, or gelatin and glycerine. Compositions for rectal administration are conveniently in the form of suppositories containing a conventional suppository base such as cocoa butter.

[0091] Suppository forms of the compounds of the application are useful for vaginal, urethral and rectal administrations. Such suppositories will generally be constructed of a mixture of substances that is solid at room temperature but melts at body temperature. The substances commonly used to create such vehicles include but are not limited to theobroma oil (also known as cocoa butter), glycerinated gelatin, other glycerides, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol. See, for example: Remington's Pharmaceutical Sciences, 16th Ed., Mack Publishing, Easton, PA, 1980, pp. 1530-1533 for further discussion of suppository dosage forms.

[0092] In some embodiments a compound of the application is coupled with soluble polymers as targetable drug carriers. Such polymers include, for example, polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxy-ethylaspartamide- phenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore, in some embodiments, a compound of the application is coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels.

[0093] A compound of the application including pharmaceutically acceptable salts, solvates and/or prodrugs thereof is suitably used on their own but will generally be administered in the form of a pharmaceutical composition in which the one or more compounds of the application (the active ingredient) is in association with a pharmaceutically acceptable carrier. Depending on the mode of administration, the pharmaceutical composition will comprise from about 0.05 wt% to about 99 wt% or about 0.10 wt% to about 70 wt%, of the active ingredient, and from about 1 wt% to about 99.95 wt% or about 30 wt% to about 99.90 wt% of a pharmaceutically acceptable carrier, all percentages by weight being based on the total composition.

[0094] A compound of the application is either used alone or in combination with other known agents useful for treating diseases, disorders or conditions that treatable by inhibition of EGFR, and those that are treatable with an EGFR inhibitor. When used in combination with other agents useful in treating diseases, disorders or conditions treatable by inhibition of EGFR, it is an embodiment that a compound of the application is administered contemporaneously with those agents. As used herein, "contemporaneous administration" of two substances to a subject means providing each of the two substances so that they are both active in the individual at the same time. The exact details of the administration will depend on the pharmacokinetics of the two substances in the presence of each other, and can include administering the two substances within a few hours of each other, or even administering one substance within 24 hours of administration of the other, if the pharmacokinetics are suitable. Design of suitable dosing regimens is routine for one skilled in the art. In particular embodiments, two substances will be administered substantially simultaneously, i.e., within minutes of each other, or in a single composition that contains both substances. It is a further embodiment of the present application that a combination of agents is administered to a subject in a non-contemporaneous fashion. In an embodiment, a compound of the present application is administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present application provides a single unit dosage form comprising one or more compounds of the application, an additional therapeutic agent, and a pharmaceutically acceptable carrier.

[0095] The dosage of a compound of the application varies depending on many factors such as the pharmacodynamic properties of the compound, the mode of administration, the age, health and weight of the recipient, the nature and extent of the symptoms, the frequency of the treatment and the type of concurrent treatment, if any, and the clearance rate of the compound in the subject to be treated. One of skill in the art can determine the appropriate dosage based on the above factors. In some embodiments, a compound of the application is administered initially in a suitable dosage that is adjusted as required, depending on the clinical response. Dosages will generally be selected to maintain a serum level of the compound of the application from about 0.01 μg/cc to about 1000 g/cc, or about 0.1 μg/cc to about 100 μg/cc. As a representative example, oral dosages of one or more compounds of the application will range between about 1 mg per day to about 1000 mg per day for an adult, suitably about 1 mg per day to about 500 mg per day, more suitably about 1 mg per day to about 200 mg per day. For parenteral administration, a representative amount is from about 0.001 mg/kg to about 10 mg/kg, about 0.01 mg/kg to about 10 mg/kg, about 0.01 mg/kg to about 1 mg/kg or about 0.1 mg/kg to about 1 mg/kg will be administered. For oral administration, a representative amount is from about 0.001 mg/kg to about 10 mg/kg, about 0.1 mg/kg to about 10 mg/kg, about 0.01 mg/kg to about 1 mg/kg or about 0.1 mg/kg to about 1 mg/kg. For administration in suppository form, a representative amount is from about 0.1 mg/kg to about 10 mg/kg or about 0.1 mg/kg to about 1 mg/kg. In an embodiment of the application, compositions are formulated for oral administration and the one or more compounds are suitably in the form of tablets containing 0.25, 0.5, 0.75, 1 .0, 5.0, 10.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 75.0, 80.0, 90.0, 100.0, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 mg of active ingredient per tablet. In embodiments of the application the one or more compounds of the application are administered in a single daily, weekly or monthly dose or the total daily dose is divided into two, three or four daily doses.

[0096] In the above, the term "a compound" also includes embodiments wherein one or more compounds are referenced.

III. Methods and Uses of the Application

[0097] The compounds of the application have been shown to be capable of inhibiting EGFR activity.

[0098] Accordingly, the present application includes a method for inhibiting EGFR in a cell, either in a biological sample or in a patient, comprising administering an effective amount of one or more compounds of the application to the cell. The application also includes a use of one or more compounds of the application for inhibiting EGFR in a cell as well as a use of one or more compounds of the application for the preparation of a medicament for inhibiting EGFR in a cell. The application further includes one or more compounds of the application for use in inhibiting EGFR in a cell.

[0099] As the compounds of the application have been shown to be capable of inhibiting EGFR protein activity, the compounds of the application are useful for treating diseases, disorders or conditions treatable by the inhibition of EGFR. Therefore the compounds of the present application are useful as medicaments. Accordingly, the present application includes a compound of the application for use as a medicament.

[00100] The present application also includes a method of treating a disease, disorder or condition treatable by inhibition of EGFR comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof.

[00101] The present application also includes a use of one or more compounds of the application for treatment of a disease, disorder or condition treatable by inhibition of EGFR as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of a disease, disorder or condition treatable by inhibition of EGFR. The application further includes one or more compounds of the application for use in treating a disease, disorder or condition treatable by inhibition of EGFR.

[00102] In an embodiment, the disease, disorder or condition is a neoplastic disorder. Accordingly, the present application also includes a method of treating a neoplastic disorder comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof. The present application also includes a use of one or more compounds of the application for treatment of a neoplastic disorder as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of a neoplastic disorder. The application further includes one or more compounds of the application for use in treating a neoplastic disorder. In an embodiment, the treatment is in an amount effective to ameliorate at least one symptom of the neoplastic disorder, for example, reduced cell proliferation or reduced tumor mass, among others, in a subject in need of such treatment.

[00103] In another embodiment of the present application, the disease, disorder or condition treatable by inhibition of EGFR is cancer. Accordingly, the present application also includes a method of treating cancer comprising administering a therapeutically effective amount of one or more compounds of the application to a subject in need thereof. The present application also includes a use of one or more compounds of the application for treatment of cancer as well as a use of one or more compounds of the application for the preparation of a medicament for treatment of cancer. The application further includes one or more compounds of the application for use in treating cancer. In an embodiment, the compound is administered for the prevention of cancer in a subject such as a mammal having a predisposition for cancer.

[00104] In an embodiment, the cancer is a solid cancer or a so-called liquid cancer, and can be selected from a cancer of the skin, blood, prostate, colorectum, pancreas, kidney, ovary, breast, for example mammary, liver, tongue and lung. In another embodiment, the cancer is selected from leukaemia, lymphoma, non-Hodgkin's lymphoma and multiple myeloma. The cancer target includes particularly those for which regulatory approval has already been granted for other EGFR inhibitors. These cancers include colorectal cancer, head and neck cancer, pancreatic cancer, non-small cell lung cancer, and glioma.

[00105] In an embodiment, the disease, disorder or condition is a disease, disorder or condition associated with an uncontrolled and/or abnormal cellular activity affected directly or indirectly by alteration of EGFR protein activity. In another embodiment, the uncontrolled and/or abnormal cellular activity that is affected directly or indirectly by altered EGFR activity is proliferative activity in a cell. Accordingly, the application also includes a method of inhibiting proliferative activity in a cell, comprising administering an effective amount of one or more compounds of the application to the cell. The present application also includes a use of one or more compounds of the application for inhibition of proliferative activity in a cell as well as a use of one or more compounds of the application for the preparation of a medicament for inhibition of proliferative activity in a cell. The application further includes one or more compounds of the application for use in inhibiting proliferative activity in a cell.

[00106] The present application also includes a method of inhibiting uncontrolled and/or abnormal cellular activities affected directly or indirectly by EGFR protein in a cell, either in a biological sample or in a subject, comprising administering an effective amount of one or more compounds of the application to the cell. The application also includes a use of one or more compounds of the application for inhibition of uncontrolled and/or abnormal cellular activities affected directly or indirectly by EGFR protein in a cell as well as a use of one or more compounds of the application for the preparation of a medicament for inhibition of uncontrolled and/or abnormal cellular activities affected directly or indirectly by EGFR protein inhibition in a cell. The application further includes one or more compounds of the application for use in inhibiting uncontrolled and/or abnormal cellular activities affected directly or indirectly by EGFR.

[00107] Accordingly, the present application also includes a method of treating a disease, disorder or condition that is treatable by inhibition of EGFR comprising administering a therapeutically effective amount of one or more compounds of the application in combination with another known agent useful for such treatment. The present application also includes a use of one or more compounds of the application in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibition of EGFR for treatment of a disease, disorder or condition treatable by inhibition of EGFR as well as a use of one or more compounds of the application in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibition of EGFR, for the preparation of a medicament for treatment of a disease, disorder or condition treatable by inhibition of EGFR. The application further includes one or more compounds of the application in combination with another known agent useful for treatment of a disease, disorder or condition treatable by inhibition of EGFR for use in treating a disease, disorder or condition treatable by inhibition of EGFR. In an embodiment, the disease, disorder or condition treatable by inhibition of EGFR is a cancer such as multiple myeloma, lymphoma, leukemia, ovarian cancer, brain cancer, lung cancer, and pancreatic cancer. Treatable EGFR-mediated cancers thus include benign or malignant tumors (e.g., renal, liver, kidney, bladder, breast, gastric, ovarian, colorectal, prostate, pancreatic, lung, vulva, and thyroid); hepatic carcinomas; sarcomas; glioblastomas; and various head and neck tumors including particularly head and neck cancers and especially squamous cell carcinoma of the head and neck, colorectal cancers, gastrointestinal cancers, brain tumours including glioblastomas, and tumours of the lung including non-small-cell lung carcinoma, and of the breast, pancreas, esophagus, kidney, ovary, cervix and prostate.

[00108] In a further embodiment, the disease, disorder or condition treatable by inhibition of EGFR is cancer and the one or more compounds of the application are administered in combination with one or more additional cancer treatments. In another embodiment, the additional cancer treatment is selected from radiotherapy, chemotherapy, targeted therapies such as antibody therapies and small molecule therapies such as tyrosine-kinase inhibitors, immunotherapy, hormonal therapy and anti-angiogenic therapies.

EXAMPLES

[00109] The following non-limiting examples are illustrative of the present application: Chemical Synthesis

A. General methods

[00110] All starting materials used herein were commercially available or earlier described in the literature. The ¹ H and ¹³C NMR spectra were recorded either on Bruker 300, Bruker DPX400 or Varian +400 spectrometers operating at 300, 400 and 400 MHz for ¹ H NMR respectively, using TMS or the residual solvent signal as an internal reference, in deuterated chloroform as solvent unless otherwise indicated. All reported chemical shifts are in ppm on the delta- scale, and the fine splitting of the signals as appearing in the recordings is generally indicated, for example as s: singlet, br s: broad singlet, d: doublet, t: triplet, q: quartet, m: multiplet. Unless otherwise indicated, in the tables below, ¹ H NMR data was obtained at 400 MHz, using CDC as the solvent. HPLC and LCMS data were collected using an Agilent 1200 HPLC with MS detector using standard reversehase chromatography conditions.

[00111] Purification of products was carried out using Chem Elut Extraction Columns (Varian, cat #1219-8002), Mega BE-SI (Bond Elut Silica) SPE Columns (Varian, cat # 12256018; 12256026; 12256034) or by flash chromatography in silica-filled glass columns.

Representative synthesis of compounds of Formula I

[00112] Scheme VI below outlines the synthesis of compounds of Formula I, represented by 1-1 , when R¹ is 3-chloro-2-fluoro-phenyl and R² and R³ are difluoromethoxyethyl.

1-1

a.BnBr, K2CO3, DMF b. Ac₂0, HNO3 c.Fe powder, AcOH d. formamidine acetate, iBuOH e. POC , NEt3 f. 3-chloro-2-fluoro-phenol, K2CO3, DMF g. TFA, reflux h. 2-(difluoromethoxy)ethyl 4-methylbenzenesulfonate, K2CO3, DMF.

Scheme VI

[00113] Scheme VII below outlines the synthesis of compounds of Formula I, represented by I-2 where R¹ is 3-chloro-2-fluoro-phenyl, R³ is methyl and R² is difluoromethoxyethyl.

KK 1-2

K2CO3, DMF b. TFA, reflux C. 2-(difluoromethoxy)ethyl ethylbenzenesulfonate, K2CO3, DMF

Scheme VII

Exa -dibenzyloxybenzoate (BB)

[00114] To a stirred solution of 3,4-dihydroxybenzoic acid (10 g, 64.88 mmol) in DMF (100 mL) was added potassium carbonate (35.86 g, 259.5 mmol) followed by benzyl bromide (23.89 mL, 201 .1 mmol). The resulting suspension was stirred at room temperature overnight. The mixture was diluted with ethyl acetate and washed with brine (1x), brine:water (2x) and brine (1 x). The organic phase was dried, filtered and concentrated in vacuo giving an amber oil which slowly solidified. The residue was stirred in hexanes and filtered to give the desired product as a white solid (22g, 81 %). [See Australian Journal of Chemistry, 70(1 ), 52-60; 2017].

Example 2: Benzyl 4,5-dibenzyloxy-2-nitro-benzoate (CC)

[00115] 3,4-dibenzyloxybenzoate (47.7 g, 1 12.4 mmol) was stirred in acetic anhydride (300 mL) until all of the material was dissolved. The reaction vessel was placed in a water bath to maintain the temperature; the mixture was then treated with fuming nitric acid (9.38 ml_, 224.7 mmol) over a period of 2 min. After approximately 5 min a precipitate formed. The mixture was quenched via the addition of ice and water (-500 ml_ total) and stirred vigorously. The resulting suspension was filtered and the filter cake was stirred in water. The suspension was filtered; the filter cake was dissolved in DCM and the residual water was separated. The organic phase was dried, filtered and concentrated in vacuo then stirred in hexanes. The resulting suspension was filtered to give the desired product as a white solid (52 g, quantitative). ¹ H NMR (400 MHz, d6-DMSO) δ 7.78 (s, 1 H), 7.49-7.32 (m, 16H), 5.33 (s, 2H), 5.31 (s, 2H), 5.29 (s, 2H).

Example 3: Benzyl 2-amino-4,5-dibenzyloxy-benzoate (DD)

[00116] To a stirred suspension of benzyl 4,5-dibenzyloxy-2-nitro- benzoate (52 g, 1 10.8 mmol) in acetic acid (100 ml_), water (5 ml_) and THF (100 ml_) at 65 °C was added iron (I II) chloride (400 mg), followed by iron powder (34 g). The resulting mixture was stirred at 65 °C for 3 h (starting material dissolves, mixture turns dark. The mixture was filtered through a pad of celite; the filtrate was diluted with ethyl acetate (and minimal THF to aid dissolution) then washed with brine (2x), NaHC03 (3x) and brine (1x). The organic phase was dried, filtered and concentrated then passed through a plug of silica gel eluting with ethyl acetate. The eluent was concentrated in vacuo, stirred in diethyl ether and filtered to give the desired product as an off-white solid (37 g, 77 %). ¹ H NMR (400 MHz, d6-DMSO) δ 7.50-7.29 (m, 17H), 6.49 (s, 2H), 5.24 (s, 2H), 5.13 (s, 2H), 4.96 (s, 2H).

-dibenzyloxy-3H-quinazolin-4-one (EE)

[00117] Benzyl 2-amino-4,5-dibenzyloxy-benzoate (37 g, 84.18 mmol) was stirred in isobutyl alcohol (200 ml_) at 95 °C. When complete dissolution was observed the dark solution was treated with formamidine acetate (12.3 g, 1 17.9 mmol) then stirred at 95 °C for 2 h (suspension was observed after -10 min). The mixture was cooled to room temperature and filtered. The filter cake was stirred in diethyl ether and filtered to give the desired product as a white solid (21 g, 70 %). ¹ H NMR (400 MHz, d6-DMSO) δ 7.94 (s, 1 H), 7.58 (s, 1 H), 7.53-7.45 (m, 4H), 7.44-7.36 (m, 4H), 7.36-7.28 (m, 2H), 7.23 (s, 1 H), 5.31 (s, 2H), 5.26 (s, 2H).

Exa -dibenzyloxy-4-chloro-quinazoline (FF/D)

[00118] To a stirred suspension of 6,7-dibenzyloxy-3H-quinazolin-4-one (30.9 g, 86.2 mmol) in DCE (200 ml_) was added POC (15.8 ml_, 172.4 mmol) followed by trimethylamine (36 ml_, 258.6 mmol). The resulting mixture was stirred at 80 °C for 1 .5 h (TLC analysis showed reaction was complete). The mixture was cooled in an ice bath then quenched via the addition of ice and water. The mixture was diluted with DCM and filtered through celite. The organic layer was separated and washed with brine, then dried with anhydrous magnesium sulfate and some silica gel. The mixture was filtered and concentrated then triturated with ether and hexanes to give the desired product as a beige solid (27 g, 83 %), used directly in the subsequent step.

Example 6(a): 6-benzyloxy-4-(3-bromo-2-fluoro-phenoxy)-7-methoxy- quinazoline (JJ-1 )

[00119] To a stirred solution of 6-benzyloxy-4-chloro-7-methoxy- quinazoline (2 g, 6.65 mmol) and potassium carbonate (2.29 g, 16.63 mmol) in DMF (10 mL) was added 3-bromo-2-fluoro-phenol (1 .52 g, 7.98 mmol) and the resulting mixture was stirred at 90 °C for 1 h. The mixture was cooled to room temperature and diluted with water, diethyl ether and hexanes. The suspension was filtered and washed with water; the filter cake was stirred in diethyl ether then filtered to collect the desired product as a beige solid which was dried in the oven (2.90 g, 96 %). H NMR (400 MHz, DMSO-de) ¹H NMR (400 MHz, DMSO-de) δ 8.61 (s, 1 H), 7.75 (s, 1 H), 7.72 (t, J = 8.0 Hz, 1 H), 7.60 - 7.51 (m, 3H), 7.47 (s, 1 H), 7.46 - 7.36 (m, 3H), 7.32 (t, J = 8.2 Hz, 1 H), 5.33 (s, 2H), 4.02 (s, 3H).

[0002] The following compounds were made following a similar procedure:

Example 7(a): 4-(3-bromo-2-fluoro-phenoxy)-7-methoxy-quinazolin-6-ol trifluoroacetate (KK-1 )

[00120] 6-benzyloxy-4-(3-bromo-2-fluoro-phenoxy)-7-methoxy- quinazoline (2.89 g, 6.35 mmol) was stirred in TFA (15 mL) and anisole (1 mL) at 70 °C for 3 h. The mixture was concentrated in vacuo and stirred with diethyl ether; the resulting suspension was filtered to give the desired product as a pale solid (2.90 g, 95%). ¹H NMR (400 MHz, DMSO-de) δ 10.48 (bs, 1 H), 8.54 (s, 1 H), 7.70 (ddd, J = 8.0, 6.2, 1 .6 Hz, 1 H), 7.55 (ddd, J = 8.5, 7.2, 1 .6 Hz, 1 H), 7.52 (s, 1 H), 7.41 (s, 1 H), 7.31 (td, J = 8.2, 1 .5 Hz, 1 H), 4.02 (s, 3H).

[0003] The following compounds were made following a similar procedure:

Example 8(a): 4-(3-bromo-2-fluoro-phenoxy)-6-[2- (difluoromethoxy)ethoxy]-7-methoxy-quinazoline (I-3)

[00121] To a stirred suspension of 4-(3-bromo-2-fluoro-phenoxy)-7- methoxy-quinazolin-6-ol trifluoroacetate (2.90 g, 6.05 mmol) and potassium carbonate (3.34 g, 24.21 mmol) in DMF (20 ml_) was added 2- (difluoromethoxy)ethyl 4-methylbenzenesulfonate (2.42 g, 9.08 mmol); the resulting mixture was stirred at 70 °C for 3 h. The mixture was then diluted with ethyl acetate and washed with brine (2x), water (1x) and brine (1x). The organic phase was dried, filtered and concentrated in vacuo then chromatographed in 20 - 80 % ethyl acetate in hexanes. The product containing fractions were concentrated in vacuo then stirred in hexanes and diethyl ether. The resulting suspension was filtered to give the desired product as a white solid. ¹H NMR (400 MHz, DMSO-de) δ 8.60 (s, 1 H), 7.71 (ddd, J = 8.0, 6.1 , 1 .6 Hz, 1 H), 7.64 (s, 1 H), 7.55 (ddd, J = 8.2, 7.1 , 1 .6 Hz, 1 H), 7.46 (s, 1 H), 7.32 (td, J = 8.2, 1 .6 Hz, 1 H), 6.79 (t, J = 75 Hz, 1 H), 4.47 - 4.40 (m, 2H), 4.30 - 4.23 (m, 2H), 4.02 (s, 3H).

[0004] The following compounds were made following a similar procedure:

Example 9: 2-[3-[6-[2-(difluoromethoxy)ethoxy]-7-methoxy-quinazolin-4- yl]oxy-2-fluoro-phenyl]ethynyl-trimethyl-silane (0-1 )

[00122] To a stirred solution of 4-(3-bromo-2-fluoro-phenoxy)-6-[2- (difluoromethoxy)ethoxy]-7-methoxy-quinazoline (1 g, 2.18 mmol) in dioxane (5 ml_) was added trimethylamine (1.52 ml_, 10.89 mmol) and ethynyl(trimethyl)silane (3.07 ml_, 21 .78 mmol); the flask was purged with nitrogen. The resulting solution was treated with copper (I) iodide (41 .4 mg, 0.218 mmol) and bis(triphenylphosphine)palladium(ll) dichloride (153 mg, 0.218 mmol), the flask was sealed and the mixture as stirred at 80 °C overnight. The mixture was diluted with ethyl acetate and washed with NaHC03 (1x) and brine (1x). The organic phase was dried, filtered and concentrated onto silica gel then chromatographed in 20-60% ethyl acetate in hexanes. The product containing fractions were concentrated in vacuo giving a sticky foam (800 mg, 78%). ¹ H NMR (400 MHz, DMSO-de) δ 8.60 (s, 1 H), 7.64 (s, 1 H), 7.59 (ddd, J = 7.9, 6.2, 1 .6 Hz, 1 H), 7.51 (ddd, J = 7.9, 6.2, 1 .6 Hz, 1 H), 7.46 (s, 1 H), 7.33 (t, J = 8.0 Hz, 1 H), 6.79 (t, J = 76 Hz, 1 H), 4.46 - 4.40 (m, 2H), 4.29 - 4.24 (m, 2H), 4.02 (s, 3H), 0.25 (s, 9H).

Example 10: 6-[2-(difluoromethoxy)ethoxy]-4-(3-ethynyl-2-fluoro- phenoxy)-7-methoxy-quinazoline (I-4)

[00123] To a stirred solution of 2-[3-[6-[2-(difluoromethoxy)ethoxy]-7- methoxy-quinazolin-4-yl]oxy-2-fluoro-phenyl]ethynyl-trimethyl-silane (800 mg, 1 .68 mmol) in methanol (5 ml_) was added potassium carbonate (926 mg, 6.71 mmol); stirred for 1 h at room temperature. The mixture was diluted with ethyl acetate and washed with brine (1x), water (1x) and brine (1x). The organic phase was dried, filtered and concentrated in vacuo then chromatographed in 0-50% ethyl acetate in hexanes. The product containing fractions were concentrated in vacuo then stirred in hexanes. The suspension was filtered to give the desired product as a white solid (368 mg, 54%). ¹H NMR (400 MHz, DMSO-de) δ 8.60 (s, 1 H), 7.64 (s, 1 H), 7.61 - 7.51 (m, 2H), 7.45 (s, 1 H), 7.34 (t, J = 8.0 Hz, 1 H), 6.79 (t, J = 76 Hz, 1 H), 4.63 (s, 1 H), 4.48 - 4.38 (m, 2H), 4.31 - 4.20 (m, 2H), 4.02 (s, 3H). Example 11 (a): 6,7-dibenzyloxy-4-(3-bromo-2-fluoro- phenoxy)quinazoline

[00124] To a stirred suspension of 6,7-dibenzyloxy-4-chloro-quinazoline and potassium carbonate (2.1 g, 5.6 mmol) and potassium carbonate (1 .93 g, 14 mmol) in DMF (28 mL) was added 3-bromo-2-fluoro-phenol (1 .18 g, 6.2 mmol) and the resulting mixture was stirred at 100 °C for 1 h. The mixture was diluted with ethyl acetate and washed with brine (2x). The organic phase was dried, filtered and concentrated in vacuo then stirred in diethyl ether overnight. The resulting suspension was filtered to collect the desired product as a pale solid (2.43 g, 82%). H NMR (400 MHz, DMSO-de) δ 8.59 (s, 1 H), 7.77 (s, 1 H), 7.71 (ddd, J = 8.0, 6.1 , 1 .5 Hz, 1 H), 7.58 - 7.50 (m, 6H), 7.48 - 7.40 (m, 4H), 7.39 - 7.28 (m, 3H), 5.44 (s, 2H), 5.38 (s, 2H).

[0005] The following compounds were made following a similar procedure:

¹ H NMR (d6-DMSO, 400 MHz) δ 8.58 (s, 1 H), 7.76 (s, 1 H), 7.68-7.46 (m,

7H), 7.46-7.32 (m, 7H), 5.42 (s, 2H), 5.37 (s, 2H).

Example 12(a): 4-(3-bromo-2-fluoro-phenoxy)quinazoline-6,7-diol

trifluoroacetate (HH-1 )

[00125] 6,7-dibenzyloxy-4-(3-bromo-2-fluoro-phenoxy)quinazoline (2.3 g, 4.3 mmol) was stirred in TFA (15 ml_) at 70 °C overnight. The mixture was concentrated in vacuo; the residue was stirred in DCM and concentrated in vacuo (2x). The crude product was used directly in the subsequent reaction.

[0006] The following compounds were made following a similar procedure:

Example 13(a): 4-(3-bromo-2-fluoro-phenoxy)-6,7-bis[2- (difluoromethoxy)ethoxy]quinazoline (I-5)

[00126] To a stirred suspension of 4-(3-bromo-2-fluoro- phenoxy)quinazoline-6,7-diol trifluoroacetate (2.0 g, 4.3 mmol) and potassium carbonate (2.97 g, 22 mmol ) in DMF (30 ml_) was added 2- (difluoromethoxy)ethyl 4-methylbenzenesulfonate (2.86 g, 1 1 mmol); the resulting mixture was stirred at 80 °C for 2 h. The mixture was diluted with water and extracted with ethyl acetate (3x). The combine organics were washed with water (1 x) and brine (2x). The organic phase was dried, filtered and concentrated in vacuo then chromatographed in 20-50% ethyl acetate in hexanes. Product containing fractions were concentrated in vacuo and triturated with diethyl ether and hexanes. The resulting suspension was stirred overnight and filtered to give the desired product as a white solid (1 .72 g, 74%).

1 H NMR (400 MHz, DMSO-de) δ 8.61 (s, 1 H), 7.71 (ddd, J = 7.9, 6.2, 1 .6 Hz, 1 H), 7.67 (s, 1 H), 7.56 (ddd, J = 8.5, 7.1 , 1 .6 Hz, 1 H), 7.52 (s, 1 H), 7.32 (td, J = 8.2, 1.6 Hz, 1 H), 6.78 (t, J = 76.0 Hz, 1 H), 6.77 (t, J = 76.0 Hz, 1 H), 4.52 - 4.39 (m, 4H), 4.32 - 4.23 (m, 4H).

[0007] The following compounds were made following a similar procedure:

Yield

Ex.# Cpd.# Structure Nomenclature Appearance

(%)

4-(3-bromophenoxy)-6,7-

(I-6) bis[2- 3(b) A White solid 67

(difluoromethoxy)ethoxy]qu

F inazoline ¹ H NMR (d6-DMSO, 400 MHz) δ 8.60 (s, 1 H), 7.66-7.62 (m, 1 H), 7.63 (s, 1 H), 7.57-7.52 (m, 1 H), 7.50-7.44 (m, 1 H), 7.47 (s, 1 H), 7.42-7.37 (m, 1 H), 6.78 (2t, J = 76 Hz, 2H), 4.48-4.39 (m, 4H), 4.31 -4.25 (m, 4H).

4-(3-chloro-2-fluoro- phenoxy)-6,7-bis[2-

White solid 50 (difluoromethoxy)ethoxy]qu

(1-1 )

inazoline

(c)

¹ H NMR (d6-DMSO, 400 MHz) δ 8.61 (s, 1 H), 7.68 (s, 1 H), 7.64-7.58 (m, 1 H), 7.57-7.51 (m, 1 H), 7.51 (s, 1 H), 7.41 -7.35 (m, 1 H), 6.79 (t, J = 76 Hz, 1 H), 6.78 (t, J = 76 Hz, 1 H),4.50-4.42 (m, 4H), 4.31 -4.24 (m, 4H).

Example 14(a): 2-[3-[6,7-bis[2-(difluoromethoxy)ethoxy]quinazolin-4- yl]oxy-2-fluoro-phenyl]ethynyl-trimethyl-silane (O)

[00127] 4-(3-bromo-2-fluoro-phenoxy)-6,7-bis[2- (difluoromethoxy)ethoxy]quinazoline (1 .0 g, 1 .9 mmol), bis(triphenylphosphine)palladium(ll) dichloride (133 mg, 0.19 mmol) and copper (I) iodide (36 mg, 0.19 mmol) were charged to an oven dried RBF with a stir bar; flask was purged with nitrogen. Dioxane (8.5 mL) was added, followed by trimethylamine (1 .3 mL, 9.3 mmol) and ethynyl(trimethyl)silane (2.69 mL, 19 mmol) and the resulting mixture was stirred at 80 °C overnight.

The mixture was filtered through a pad of celite, concentrated onto silica gel then chromatographed in 40-50% ethyl acetate in hexanes. The product containing fractions were concentrated in vacuo giving the desired product (1 .01 g, 98%). H NMR (d6-DMSO, 400 MHz) δ 8.60 (s, 1 H), 7.67 (s, 1 H), 7.59- 7.49 (m, 2H), 7.50 (s, 1 H), 7.38-7.28 (m, 1 H), 6.77 (2t, J = 78 Hz, 1 H), 4.52- 4.39 (m, 4H), 4.32-4.22 (m, 4H), 0.26 (s, 9H).

[00128] The following compounds were made following a similar procedure: Yield

Ex.# Cpd.# Structure Nomenclature Appearance

(%)

2-[3-[6,7-bis[2-

(difluoromethoxy)ethoxy]qu

inazolin-4- Clear oil quant yl]oxyphenyl]ethynyl-

(0-3)

4(b) trimethyl-silane

¹ H NMR (d6-DMS0, 400 MHz) δ 8.60 (s, 1 H), 7.64 (s, 1 H), 7.54-7.37 (m, 5H), 6.78 (2t, J = 78 Hz, 2H), 4.48-4.39 (m, 4H), 4.31 -4.24 (m, 4H), 0.24 (s, 9H).

Example 15(a): 6,7-bis[2-(difluoromethoxy)ethoxy]-4-(3-ethynyl-2-fluoro- phenoxy)quinazoline (I-7)

[00129] To a stirred solution of 2-[3-[6,7-bis[2- (difluoromethoxy)ethoxy]quinazolin-4-yl]oxy-2-fluoro-phenyl]ethynyl-trimethyl- silane (1.0 g, 1 .8 mmol) in methanol (18 mL) was added potassium carbonate

(0.99 g, 7.2 mmol). The resulting suspension was stirred at room temperature for 45 min. The mixture was diluted with ethyl acetate and washed with water (1x) and brine (1x). The organic phase was dried, filtered and concentrated in vacuo then chromatographed in 30-50% ethyl acetate in hexanes. Product containing fractions were concentrated in vacuo giving the desired product (210 mg, 24%).¹ H NMR (d6-DMSO, 400 MHz) δ 8.60 (s, 1 H), 7.67 (s, 1 H), 7.62-7.51 (m, 2H), 7.50 (s, 1 H), 7.39-7.30 (m, 1 H), 6.78 (2t, J = 78 Hz, 2H), 4.61 (s, 1 H), 4.50-4.40 (m, 4H), 4.32-4.22 (m, 4H).

[00130] The following compounds were made following a similar procedure: Yield

Ex.# Cpd.# Structure Nomenclature Appearance

(%)

6,7-bis[2-

(difluoromethoxy)ethoxy]- 4-(3- White solid 64

(I-8)

5(b) F ethynylphenoxy)quinazolin

e

H NMR (d6-DMSO, 400 MHz) δ 8.59 (s, 1 H), 7.65 (s, 1 H), 7.55-7.40 (m, 5H), 6.78 (2t, J = 78 Hz, 2H), 4.48-4.39 (m, 4H), 4.31 -4.22 (m, 5H).

Example 16: [4-(3-chloro-2-fluoro-phenoxy)-7-methoxy-quinazolin-6-yl]

-[2-(difluoromethoxy)ethyl]-N-methyl-carbamate hydrochloride (I-9)

[00131] 2-(difluoromethoxy)-N-methyl-ethanamine hydrochloride (162 mg, 1 .0 mmol) was stirred in DCM (5 ml_) at 0 °C and treated with triphosgene (297 mg, 1 .0 mmol), followed by pyridine (317 mg, 4.0 mmol). After 5 h, the mixture was concentrated in vacuo and mixed with 4-(3-chloro-2-fluoro- phenoxy)-7-methoxy-quinazolin-6-ol trifluoroacetate (291 mg, 0.67 mmol) and potassium carbonate (276 mg, 2.0 mmol) in DMF (5 ml_). The resulting mixture was stirred at room temperature overnight. The mixture was then diluted with ethyl acetate and washed with NaHCCte (1x) and brine (1 x). The organic phase was dried, filtered and concentrated in vacuo then stirred in diethyl ether. The resulting suspension was filtered to remove starting material. The filtrate was diluted with ether and washed with water (2x) and brine (1 x). The ether layer was dried, filtered and treated with HCI, 2M in diethyl ether (0.25 ml_). The resulting suspension was filtered to give the desired product as a white solid (225 mg, 66%). H NMR (400 MHz, DMSO-de) δ 8.71 (s, 1 H), 8.08 (s, 1 H), 7.65-7.51 (m, 2H), 7.58 (s, 1 H), 7.42-7.34 (m, 1 H), 6.77 (t, J = 76 Hz, 1 H), 4.18- 3.99 (m, 2H), 4.02 (s, 3H), 3.76-3.55 (m, 4H), 2.99 (s, 3H). Biological Testing

Example 17: Determination of hERG activity by Manual Patch-Clamp

Cell lines and cell culture

[00132] HEK 293 cell line stably expressing hERG channel (Cat. K1236) was purchased from Invitrogen. The cells were cultured in medium containing of 85% DMEM, 10% dialyzed FBS, 0.1 mM NEAA, 25mM HEPES, 100U/ml_ Penicillin-Streptomycin and 5μg/mL Blasticidin and 400μg/mL Geneticin. Cells were split using TrypLE™ Express about three times a week, and maintained between -40% to -80% confluence. Before the assay, the cells were seeded onto the coverslips at 5 χ 105cells /per 6 cm cell culture dish and induced with doxycycline at 1 μg/ml for 48 hours.

Solution preparations

[00133] 1 ) Extracellular solution (in mM): 132 NaCI, 4 KCI, 3 CaCI₂, 0.5 MgCI₂, 1 1 .1 glucose, and 10 HEPES (pH adjusted to 7.35 with NaOH)

[00134] 2) Intercellular solution (in mM): 140 KCI, 2 MgCI₂, 10 EGTA, 5 Mg ATP and 10 HEPES (pH adjusted to 7.35 with KOH)

The working solution preparation for test article

[00135] 1 ) Test compounds were initially prepared in DMSO with final concentration of 30 mM as stock solution according to SOP-ADMET-MAN-007.

[00136] 2) Then stock solution of each compound was serial-diluted in ratio of 1 :3 with DMSO to prepare additional 4 intermediate solutions concentrated at 10, 3.33, 1 .1 1 , 0.37 mM, respectively.

[00137] 3) Before hERG measurement, the working solutions were finally prepared by dilution of above described serial solutions in 1000 folds using extracellular solution.

[00138] 4) hERG current inhibition in presence of 5 doses, consisting of 30, 10, 3.33, 1 .1 1 and 0.37 μΜ, were tested for ICso determination.

Data acceptance criteria

[00139] The following criteria were used to determine data acceptability. 1 ) Initial seal resistances GQ; 2) Access resistance <10 ΜΩ and a series resistance voltage error < 5 mV;

3) Stable leakage current <100 pA at a test potential;

4) Normal test pulse current waveform (e.g., hERG peak tail) current amplitude greater than prepulse current amplitude and the peak tail amplitude >250 pA;

5) Apparent rundown of test pulse current amplitude <2% per min.

Data analysis

[00140] Data that met the above criteria for hERG current quality were further analyzed as described in the following steps.

1 ) Percent current inhibition was calculated using the following equation.

1 ; ; X 100

Peak tail current_vehicle )

Note: Patch Master or Clampfit software was used to extract the peak current from the original data.

2) The dose response curve of test articles was plotted with percentage of hERG inhibition against the concentration of test articles using Graphpad Prism 5.0, and the data was fit to a sigmoid dose-response curve with a variable slope.

Results

[00141] The ICso values for representative compounds of Formula I are presented in table 1 . Representative compounds of the application have IC50 values that are approximately 1 order of magnitude greater than their corresponding nitrogen counterparts (disclosed in WO2016/123706 A1 ) and therefor these compounds have a significantly lower risk for cardiovascular side effects. Table 1

Example 18: Determination of Kinase Specificity against a Panel of 468 Kinases

Protocol

[00142] For most assays, kinase-tagged T7 phage strains were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage from a frozen stock (multiplicity of infection = 0.4) and incubated with shaking at 32°C until lysis (90- 150 minutes). The lysates were centrifuged (6,000 x g) and filtered (0.2μιη) to remove cell debris. The remaining kinases were produced in HEK-293 cells and subsequently tagged with DNA for qPCR detection. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands for 30 minutes at room temperature to generate affinity resins for kinase assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1 % BSA, 0.05 % Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific phage binding. Binding reactions were assembled by combining kinases, liganded affinity beads, and test compounds in 1x binding buffer (20 % SeaBlock, 0.17x PBS, 0.05 % Tween 20, 6 mM DTT). Test compounds were prepared as 40x stocks in 100% DMSO and directly diluted into the assay. All reactions were performed in polypropylene 384-well plates in a final volume of 0.04 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (1x PBS, 0.05 % Tween 20). The beads were then re- suspended in elution buffer (1x PBS, 0.05 % Tween 20, 0.5 μΜ non-biotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. The kinase concentration in the eluates was measured by qPCR.

Results & Discussion

[00143] The hydrochloride salt of compound 1-1 (1-1 -HCI) was assessed against a panel of 486 kinases, including WT EGFR, EGFR mutant and ephrin receptor tyrosine kinases. Ultrasensitive quantitative PCR (qPCR) was used to measure levels of immobilized kinases after treatment with 1-1 -HCI @ 300 nM. 1-1 -HCI exhibited inhibitory activity against WT and certain mutant EGFR kinases containing activating mutations, such as the E746-A750del, L747- E749del/A750P and L747-S752del/P753S mutants (see Table 2). The compound proved ineffective against the T790M mutant as well as related kinases ERBB2 and ERBB3.

Table 2 DiscoverX KINOMEscan results

ERBB2 ERBB2 100

ERBB4 ERBB4 98

LCK (Lymphocyte TK ) LCK 80

EPHA6 EPHA6 2.8

Example 19: Evaluation of 1-1 Permeability in Caco-2 Cells

[00144] P-glycoprotein (Pgp) is a member of the ABC-transporter family that transports substances across cellular membranes acting as an energy- dependent efflux pump extruding drugs out of the cells. Increased expression of Pgp in cancer cells is one of the major mechanisms of cancer resitances and chemotherapy and thus Pgp plays a key role on the pharmacokinetics of drug absorption and distribution.

Protocol

[00145] 50 μΙ_ and 25 mL of cell culture medium were added to each well of the Transwell insert and reservoir, respectively. And then the HTS transwell plates were incubated at 37 °C, 5% C02 for 1 hour before cell seeding.

[00146] Caco-2 cells were diluted to 6.86x10⁵ cells/mL with culture medium and 50 μΙ_ of cell suspension were dispensed into the filter well of the 96-well HTS Transwell plate. Cells were cultivated for 14-18 days in a cell culture incubator at 37 °C, 5% C02, 95% relative humidity. Cell culture medium was replaced every 2 days, beginning no later than 24 hours after initial plating.

[00147] The After 14-18 days incubation, the plates were removed from incubator. The inserts were washed twice with pre-warmed HBSS (10 mM HEPES, pH 7.4) and placed into receiver plates. 75 μΙ_ and 235 μΙ_ of buffer were added to each Transwell inserts and receiver wells, respectively. Then the plates were incubated for 30 min at 37 °C with shaking at 150 rpm.

[00148] TEER of each well was determined. The TEER value of each well should be greater than 230 ohms- cm².

[00149] TEER value (ohm- cm²) = TEER measurement (ohms) x Area of membrane (cm²) [00150] Stock solutions of 1-1 and control compounds were prepared in DMSO at 10 mM and then diluted to 1 μΜ with HBSS (10 mM HEPES, pH 7.4,) to get compound working solution. To determine the rate of drug transport in the apical to basolateral direction, 75 μΙ_ of compound working solution was added to the filter well (apical compartment) and 235 μΙ_ of HBSS (10 mM HEPES, pH 7.4) to receiver plate (basolateral compartment). To determine the rate of drug transport in the basolateral to apical direction, 235 μΙ_ of compound working solution was added to each well of the receiver plate (basolateral compartment) and 75 μΙ_ of HBSS (10 mM HEPES, pH 7.4) to filter well (apical compartment). The assay was performed in duplicate.

[00151] Plates were incubated for 2 hours at 37 °C with shaking at 150 rpm on a rotary shaker. At the end of the transport period, aliquots of 50 μΙ_ were removed directly from the apical and basolateral wells and transferred to wells of new plates. Four volumes of cold methanol containing internal standards (IS, 100 nM Alprazolam, 200 nM Labetalol and 200 nM Diclofenac) was added into each well. Samples were centrifuged at 3,220 g for 30 minutes. Aliquots of 200 μΙ_ supernatant were used for LC-MS/MS analysis.

[00152] Solution was discarded from Transwell plates. 100 μΙ_ of Lucifer Yellow solution (100 μΜ in HBSS) and 300 μί of HBSS were added into each well of Transwell insert and receiver, respectively, for leakage determination. Plates were incubated at 37 °C for 30 minutes with shaking at 150 rpm on a rotary shaker. 80 μί aliquots from wells of apical and basolateral sides were transferred to solid black plates and the plate was read with Tecan InfiniteTM M 200 (Excitation/Emission wavelength 485 nM/ 530 nM).

[00153] LC/MS analysis was performed for all samples from this study using a Shimadzu liquid chromatograph separation system equipped with degasser DGU-20A3, solvent delivery unit LC-20AD, system controller CBM- 20A, column oven CTO-10ASVP and CTC Analytics HTC PAL System. Mass spectrometric analysis was performed using an API 5500 instrument from AB Inc (Canada) with an ESI interface. The data acquisition and control system were created using Analyst 1 .5 software from ABI Inc. Results

[00154] Activity as a Pgp substrate is an undesirable trait for an anticancer compound. Compounds that are substrates for Pgp will be more readily transported out of a cancer cell and therefore show reduced activity. In the present assay it is desirable for a compound to show an efflux ratio of between about 1 and 5. Results for representative compounds of the application are shown in Table 3. As can be seen, 1-1 exhibits a significantly low efflux ratio. In fact the efflux ratio for 1-1 is well below the desirable range for anticancer drugs.

Table 3: Permeability results of 1-1 and Erlotinib compared control drugs

Propranolol, Prazosin and Digoxin in Caco-2 cell line

Example 20: Kinase HotSpot Profiling

Reagents:

[00155] Base Reaction buffer; 20 mM Hepes (pH 7.5), 10 mM MgC , 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na₃V0₄, 2 mM DTT, 1 % DMSO

*Required cofactors are added individually to each kinase reaction

Reaction Procedure:

[00156] 1 . The indicated substrate was prepared in fresh Base Reaction Buffer. [00157] 2. Any required cofactors were added to the substrate solution above.

[00158] 3. Indicated kinase was added into the substrate solution and gently mixed.

[00159] 4. Compounds in DMSO were added into the kinase reaction mixture by Acoustic technology (Echo550; nanoliter range) and inclubated for 20 minutes at room temperature.

[00160] 5. 33P-ATP (specific activity 10 μθί/μΐ) was added into the reaction mixture to initiate the reaction.

[00161] 6. The kinase reaction was incubated for 2 hours at room temperature

[00162] 7. Reactions were spotted onto P81 ion exchange paper.

[00163] 8. Kinase activity was detected by filter-binding method.

Results & Discussion

[00164] Representative compounds of Formula I were evaluated against WT EGFR and mutant EGFR (L858R) kinases. ICso (nM) concentrations are illustrated in Table 4. This assay confirms that compounds of the application are effective inhibitors of the target EGFR receptors.

Table 4

Example 21: Plasma, Brain and Lung exposure ofl-1-HCI in Male SD Rats

[00165] The objective of this study was to determine the plasma pharmacokinetic profile and the brain penetration, lung penetration of 1-1 -HCI in fed male SD Rats after single intravenous (IV) and oral (PO) administrations.

Protocol

[00166] Male SD Rats were purchased from Si Bei Fu, Co. Ltd (Beijing, China). The animals were 6-8 weeks old with body weights of 200-300 g on the dosing date. The animals were housed in a 12-hour light/12-hour dark cycle environment and had free access to food and water. All animals were food fed prior to dosing. This study was approved by the Pharmaron Institutional Animal Care and Use Committee (IACUC).

[00167] Study Design: Total 36 male SD Rats were assigned to 3 groups as shown in the table below. 1-1 -HCI was administered once intravenously (2 mg/kg, free form) at 10 mL/kg and orally by gavage (25 mg/kg, free form) at a dose volume of 10 mL/kg, respectively. Plasma samples were collected at each time point after IV and PO administration, lung and brain samples were just collected at each time point after PO administration at 25 mg/kg (free form).

[00168] Formulation preparation :

[00169] Preparation of dosing for IV administration: Dissolved 3.13 mg of 1-1 -HCI in 0.073 mL of DMSO with vortexing 1 minute, sonification 2 minutes, and added 0.146 mL of Tween-20 with vortexing 1 minute, sonification 2 minutes, then added 14.357 mL of 0.5% ΗΡ-β-CD in saline (w/v) with vortexing 5 minutes to obtain a solution with concentration at 0.2 mg/mL of 1-1 -HCI.

[00170] Preparation of dosing for PO administration (25 mg/kg): Dissolved 37.34 mg of 1-1 -HCI in 0.070 mL of DMSO with vortexing 1 minutes, sonification 2 minutes, and added 0.139 mL of Tween-20 with vortexing 1 minutes, sonification 2 minutes, then added 13.702 mL of 0.5% ΗΡ-β-CD in DD water (w/v) with vortexing 5 minutes to obtain a suspension with concentration at 2.5 mg/mL of 1-1 -HCI. [00171] Preparation of dosing for PO administration (25 mg/kg): DOSE 1 : Dissolved 134.51 mg of 1-1 -HCI in 0.251 mL of DMSO with vortexing 1 minute, sonification 2 minutes, and added 0.501 mL of Tween-20 with vortexing 1 minute, sonification 2 minutes, then added 49.359 mL of 0.5% ΗΡ-β-CD in water (w/v) with vortexing 5 minutes to obtain a suspension with concentration at 2.5 mg/mL of 1-1 -HCI. DOSE 2: Dissolved 133.96 mg of 1-1 -HCI in 0.250 mL of DMSO with vortexing 1 minute, sonification 2 minutes, and added 0.499 mL of Tween-20 with vortexing 1 minute, sonification 2 minutes, then added 49.157 mL of 0.5% ΗΡ-β-CD in water (w/v) with vortexing 5 minutes to obtain a suspension with concentration at 2.5 mg/mL of 1-1 -HCI.

[00172] Sample collection

[00173] Blood Samples: For IV (2 mg/kg) administration, blood samples were collected from each animal at 0.083, 0.25, 0.5, 1 .5, 3, 6, 9, 12, 24 and 48 hour post-dose. For PO (25 mg/kg) administration, blood samples were collected from each animal at 0.25, 0.5, 1 .5, 3, 6, 9, 12, 24 and 48 hour post- dose. For PO (25 mg/kg) administration, blood, lung and brain samples were collected from each animal at 0.5, 1.5, 3, 6, 9, 12 and 24 hour post-dose.

[00174] Blood samples were collected from each animal via jugular vein (PO 25 mg/kg: heart puncture). These blood samples were placed into the tubes containing K2EDTA. The whole blood tubes were inverted several times and then centrifuged at 2000 g for 5 minutes at 4°C to obtain plasma. The plasma samples were stored frozen at -75±15°C until analysis.

[00175] Brain and lung samples were collected after animals being fully exsanguinated. Procedure: opened chest cavity, cut ventricle and performed a gentle IV saline flush (saline flush volume ~ 20 mL) with the animal head placed down at a 45 degree angle to facilitate blood removal. Brain and lung samples were collected at adopted time points, quick frozen in ice box. The brain and lung samples were stored frozen at -75±15 °C until analysis.

[00176] Sample treatment: All of the brain and lung samples were weighed and homogenized with water by brain weight and lung weight (g) to water volume (mL) using a ratio of 1 :3 before analysis. The actual concentration was the detected value multiplied by the dilution factor. 5 μί of each calibration standard working solution (5, 10, 20, 50, 100, 500, 1000, 5000 and 10000 ng/mL) was added to 50 μΙ_ of the blank SD rat plasma (or blank SD rat brain homogenate and lung homogenate) to achieve calibration standards of 0.5- 1000 ng/mL (0.5, 1 , 2, 5, 10, 50, 100, 500, 1000 ng/mL) in a total volume of 55 μί. Quality Control (QC) samples at 1 ng/mL (low), 2 ng/mL (low), 50 ng/mL (mid), 800 ng/mL (high) were prepared from the QC working solutions in the same way as calibration standards. 55 μί of standards, 55 μί of QC samples and 55 μί of unknown samples (50 μί of plasma or brain homogenate, lung homogenate with 5 μί 50% acetonitrile) were added to 200 μί of acetonitrile containing IS to precipitate proteins. Then the samples were vortexed for 30 sec. After centrifugation at 4°C, 4000 rpm for 15 min, 15 μί of the supernatant was injected into the LC-MS/MS system for quantitative analysis. All samples were processed on ice.

[00177] LC-MS/MS conditions: The LC-MS/MS system consisted of two Shimadzu LC-30AD pumps, a DGU-20A5R degasser, a Rack changer II and an AB API4000LC-MS/MS mass spectrometer. Chromatographic separation was performed on a Phenomenex Synergi 4μ Polar-RP (50 χ 2.00 mm) 80A column at room temperature. The mobile phase was composed of A: 5% acetonitrile (0.1 % formic acid); B: 95% acetonitrile (0.1 % formic acid). The flow rate was 0.7 mL/min. The injection volume was 15 μί. Positive mode electrospray ionization (ESI) was performed on a Turbo V® ion source to obtain a protonated ion of 1-1 -HCI and Dexamethasone (IS). A multiple reaction monitoring (MRM) method was selected for quantitative analysis. The optimized transitions were 495.99 → 334.00 and 393.40 → 373.30 for 1-1 -HCI and Dexamethasone, respectively. The instrument parameters were set as follows: ion spray voltage: 5500 V; curtain gas: 40 psi; nebulizer gas: 50 psi; turbo gas: 50 psi; collision gas: 10 psi; temperature: 450 °C.

[00178] Results are summarized in Tables 5-8. Table 5: Plasma Pharmacokinetic Parameters of 1-1 -HCI Following IV

Administration to Male SD Rats at 2 mg/kg

Table 6: Plasma Pharmacokinetic Parameters of 1-1 -HCI Following PO

Administration to Male SD Rats at 25 mg/kg

Table 7: Brain Pharmacokinetic Parameters of 1-1 -HCI Following PO

Administration to Male SD Rats at 25 mg/kg

Table 8: Lung Pharmacokinetic Parameters of 1-1 -HCI Following PO

Administration to Male SD Rats at 25 mg/kg

[00179] The ultimate test of a lead drug candidate is efficacy in an in vivo model. Throughout the drug development process, various ADME parameters are tested in isolation of one another to ascertain whether a drug can be viable in an in vivo setting. In a live animal, all ADME parameters are working in unison to elucidate a proper pharmacokinetic profile for the drug. 1-1 -HCI brain, lung and plasma distribution was evaluated after administration to male SD Rats. The Cmax ratio and AUCIast ratio of plasma: brain were 0.552 and 0.512. The Cmax ratio and AUCIast ratio of plasma: lung were 0.412 and 0.341 . Remarkably, distribution to brain and lung were more than plasma exposure. Approximately a 195 % and 293% increase in 1-1 -HCI exposure was observed in brain and lung, respectively, compared to plasma. Table 9 presents a comparison of the brain penetrations of 1-1 -HCI and Erlotinib.

Table 9

Example 22: 7-Day Toxicity Study in Male SD Rats

Protocol

[001 80] 1-1 -HCI was dissolved in 0.5% CMC/ 1 % Tween-20/ 0.5%HP- β - CD with vortexing for 5 minutes, followed by sonication for 20 minutes to prepare a uniform suspension with concentrations of 5, 10, and 20 mg/kg. 1-1 - HCI was administered to male SD rats at the aforementioned concentrations orally at a dosing frequency of 1 dose/day for 7 days. All study animals were monitored for behavior, such as mobility, food and water consumption, body weight (BW), eye/hair matting and other signs of physical decline. Animals that were observed to be in a continuing deteriorating condition were euthanized prior to death, or before reaching a comatose state. Animals exhibiting obvious signs of severe distress and/or pain were humanely sacrificed by carbon dioxide followed by cervical dislocation to ensure death.

[001 81 ] At the end of the study (Day 8), blood samples were first collected including whole blood for hematology & blood smear for histopathology and serum for blood chemistry. Then following blood sample collection, perfusion was conducted with 20 ml saline per rat prior to collecting brain samples and other organs. The brain was weighed and cut into 2 hemispheres, ½ for PK analysis and ½ fix. [00182] The organs (adrenals, heart, liver, spleen, lung, kidneys, brain (½ snap frozen and ½ fix), Gl (stomach, small and large intestine), pancreas, testes, thymus) were collected and organs weight were recorded followed by fixation of all collected tissue samples in 10% formalin. Bone marrow was collected and prepared immediately into bone marrow smear slides for later examination.

[00183] Serum samples were used for blood chemistry analysis of the following parameters: TBA, AST, ALT, ALP, ALB, TP, TBIL, DBIL, GGT, UA, UREA, CRE, TG, TCHO, HDL-C, LDL-C, FFA and CK.

Table 10 Blood Biochemistry Parameters

Category Abbreviation Definition

TBA Total bile acid

ALT Alanine aminotransferase

AST Aspartate aminotransferase

ALP Alkaline phosphatase

Liver function ALB Albumin

TP Total protein

TBIL Total bilirubin

DBIL Direct bilirubin

GGT Gamma-glutamyltransferase

UREA Urea

Kidney

UA Uric acid

function

CRE Creatinine

TG Triglycerides

TCHO Total Cholesterol

High density lipoprotein

HDL-C

Blood lipid cholesterol

Low density lipoprotein

LDL-C

cholesterol

FFA Free fatty acid

Heart CK Creatine kinase Category Abbreviation Definition function

[00184] All tissues of the organs (adrenals, heart, liver, spleen, lung, kidneys, brain (½ fix), Gl (stomach, small and large intestine), pancreas, testes, thymus) were collected and fixed in 10% buffered formalin for 48 hours and transferred into 70% ethanol. Fixed samples were dehydrated using ethanol and embedded in paraffin wax for sectioning.

[00185] Blood smears were prepared by placing a drop of blood approximately 4 mm in diameter on a glass slide and speading the droplet with the edge of a separate glass slide. Smears were air-dried, and then dipped into 100% methanol for 5 minutes to complete the fixing process.

[00186] Bone marrow smears were extracted from the femur at the level of the proximal diaphysis. Marrow was removed using curved forceps and was added to a drop of bovine serum albumin on a glass slide. The marrow was gently mixed with the albumin and spread using the edge of a separate glass slide. The smear was air-dried and fixed in 100% methanol for 5 minutes.

Results & Discussion

[00187] 1-1 -HCI exhibited no signs of significant body weight loss or any gross clinical abnormalities at the doses administered (5, 10, and 20 mg/kg, p.o).

[00188] There were no test article-related effects on organ weights and/or organ weight/body weight percentages for any of the 1-1 -HCI dosed groups when compared to vehicle treated rats.

[00189] Regarding hematology and blood chemistry, no test article- related effects were observed in any hematological parameter (Table 10). Individual animal increases or decreases were considered random variation. There were no consistent changes in serum chemistry parameters supportive of test article-related toxicological effects.

[00190] Blood smears of 1-1 -HCI treated mice (20 mg/kg) exhibited no difference in WBC differential counts as compared to control group rats treated with vehicle alone. Also, bone smears of 1-1 -HCI treated male SD rats (20 mg/kg) revealed no significant changes in mean or individual bone marrow cell proportions or M:E ratios when compared to control groups. Differences in the proportion of other cell types by cytological evaluation were not considered test article-related due to lack of dose dependency or relationship.

[00191 ] While the present application has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.

[00192] All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.

Claims

Claims:

1 . A compound of Formula I or a pharmaceutically acceptable salt and/or solvate thereof:

wherein:

R² is selected from (CH₂)nOR⁴ and C(0)X(CH₂)nOR⁴;

R³ is selected from Ci-₄alkyl, Ci-₄fluoroalkyl and (CH2)mOR⁵;

R⁴ is selected from CHF2 and CF3;

R⁵ is selected from CHF2, CF3 and CH3;

X is selected from NH, NCH3 and O;

n is selected from 1 , 2, 3 and 4; and

m is selected from 1 , 2, 3, and 4.

2. The compound of claim 1 , wherein R¹ is selected from substituted phenyl, wherein the substituents for R¹ are selected from one to three of CI, Br, F, Ci_-4alkyl, C2-4alkenyl, C2-4alkynyl, Ci-4fluoroalkyl, C2-4fluoroalkenyl and C2- 4fluoroalkynyl.

3. The compound of claim 2, wherein R¹ is selected from substituted phenyl, wherein the substituents for R¹ are selected from one to three of CI, Br, F, CHs, CFs, CH=CH₂ and C≡CH.

4. The compound of claim 3, wherein R¹ is selected from substituted phenyl, wherein the substituents for R¹ are selected from one or two of CI, Br, F, CH₃, CF₃, CH=CH₂ and C≡CH.

5. The compound of any one of claims 1 to 4, wherein the substituents on the phenyl of R¹ are in the ortho or meta positions.

6. The compound of claim 1 , wherein R¹ is selected from:

7. The compound of claim 6, wherein R¹ is F

8. The compound of any one of claims 1 to 7, wherein R⁴ is selected from CHF₂ and CF₃.

9. The compound of any one of claims 1 to 8, wherein R⁵ is selected from CHF₂, CF₃ and CH₃.

10. The compound of any one of claims 1 to 9, wherein X is O.

1 1 . The compound of any one of claims 1 to 10, wherein n is selected from 1 , 2 and 3.

12. The compound of claim 1 1 , wherein n is 2.

13. The compound of any one of claims 1 to 12, wherein m is selected from 1 , 2 and 3.

14. The compound of claim 13, wherein m is 2.

15. The compound of any one of claims 1 to 14, wherein both of R² and R³ are

16.

The compound of any one of claims 1 to 14, wherein R²

and R³ is CH₃.

17. A pharmaceutical composition comprising one or more compounds of Formula (I) of any one of claims 1 to 16, or a pharmaceutically acceptable salt, and/or solvate thereof, and a pharmaceutically acceptable carrier and/or diluent.

18. The pharmaceutical composition of claim 17, further comprising an additional therapeutic agent.

19. A method of treating one or more diseases, disorders or conditions treatable by inhibition of EGFR comprising administering an effective amount of one or more compounds of any one of claims 1 to 17, or a pharmaceutically acceptable salt, and/or solvate thereof, to a subject in need thereof.

20. The method of claim 19, wherein the disease, disorder or condition is a neoplastic disorder.